Effect of lipoarabinomannan from Mycobacterium avium subsp avium in Freund’s incomplete adjuvant on the immune response of cattle

The aim of the present study was to determine whether lipoarabinomannan (LAM), in combination with Freund’s incomplete adjuvant (FIA), was able to improve cell-mediated and antibody-mediated immune responses against ovalbumin (OVA) in cattle. Twenty-three calves were assigned to four treatment groups, which were subcutaneously immunized with either OVA plus FIA, OVA plus FIA and LAM from Mycobacterium avium subsp avium, FIA plus LAM, or FIA alone. Lymphoproliferation, IFN-γ production and cell subpopulations on peripheral blood mononuclear cells before and 15 days after treatment were evaluated. Delayed hypersensitivity was evaluated on day 57. Specific humoral immune response was measured by ELISA. Inoculation with LAM induced higher levels of lymphoproliferation and IFN-γ production in response to ConA and OVA (P < 0.05). Specific antibody titers were similar in both OVA-immunized groups. Interestingly, our results showed that the use of LAM in vaccine preparations improved specific cell immune response evaluated by lymphoproliferation and IFN-γ production by at least 50 and 25 percent, respectively, in cattle without interfering with tuberculosis and paratuberculosis diagnosis.

Effect of lipoarabinomannan from Mycobacterium avium subsp avium in Freund's incomplete adjuvant on the immune response of cattle.

The aim of the present study was to determine whether lipoarabinomannan (LAM), in combination with Freund's incomplete adjuvant (FIA), was able to improve cell-mediated and antibody-mediated immune responses against ovalbumin (OVA) in cattle. Twenty-three calves were assigned to four treatment groups, which were subcutaneously immunized with either OVA plus FIA, OVA plus FIA and LAM from Mycobacterium avium subsp avium, FIA plus LAM, or FIA alone. Lymphoproliferation, IFN-γ production and cell subpopulations on peripheral blood mononuclear cells before and 15 days after treatment were evaluated. Delayed hypersensitivity was evaluated on day 57. Specific humoral immune response was measured by ELISA. Inoculation with LAM induced higher levels of lymphoproliferation and IFN-γ production in response to ConA and OVA (P < 0.05). Specific antibody titers were similar in both OVA-immunized groups. Interestingly, our results showed that the use of LAM in vaccine preparations improved specific cell immune response evaluated by lymphoproliferation and IFN-γ production by at least 50 and 25%, respectively, in cattle without interfering with tuberculosis and paratuberculosis diagnosis.

[Leptospira interrogans serovar Pomona: detection of antigenic differences among 3 regional isolates from cattle and a reference strain]. / Leptospira interrogans serovar Pomona: detección de diferencias antigenicas entre tres aislamientos regionales de bovinos y una cepa de referencia.

Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.

Leptospira interrogans serovar Pomona: detección de diferencias antigenicas entre tres aislamientos regionales de bovinos y una cepa de referencia / Leptospira interrogans serovar Pomona: detection of antigenic differences among 3 regional isolates from cattle and a reference strain

Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.

Leptospira interrogans serovar Pomona: detección de diferencias antigenicas entre tres aislamientos regionales de bovinos y una cepa de referencia / Leptospira interrogans serovar Pomona: detection of antigenic differences among 3 regional isolates from cattle and a reference strain

Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.(AU)

Leptospira interrogans serovar Pomona: detección de diferencias antigenicas entre tres aislamientos regionales de bovinos y una cepa de referencia. / [Leptospira interrogans serovar Pomona: detection of antigenic differences among 3 regional isolates from cattle and a reference strain]

Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.

[Rectal cancer. The effect of the clinical stage and the degree of lymphatic invasion on local recurrence and survival. An analysis of 168 patients]. / Cáncer de recto. Influencia del estadio clínico y grado de invasión ganglionar en la recidiva local y en la supervivencia. Análisis de 168 pacientes.

INTRODUCTION: Surgery is the most important therapeutic means for treatment of rectal carcinoma. Nevertheless, from stage B2, relapse rates are high and it is therefore necessary to use supplemental treatments such as radiotherapy associated or not with chemotherapy OBJECTIVE: To assess the likelihood of being free of local, local and distant disease and overall and specific survival in function of clinical stage and degree of lymph node involvement among patients diagnosed with colo-rectal adenocarcinoma treated with radical surgery and radiotherapy associated or not with chemotherapy. MATERIALS AND METHODS: Since January 1990 up to December 1997, all patients with rectal adenocarcinoma treated with radical surgery and postoperative radiotherapy, with or without chemotherapy, were prospectively included in a database which was analyzed. RESULTS: The crude actuarial survival at five years was 61.1 +/- 9.2% and specific survival 64.2 +/- 9.2%. As for stages: B (84.1 +/- 10.1%) and C (49.9 +/- 3.3%) (p < 0.001). Likewise, for N0 84.1 +/- 10.1%, for N1 62.2 +/- 13.5% and for N2 13.7 +/- 22.3% (p < 0.001). The likelihood of being in complete remission for the overall patient population was 50.2 +/- 9.2%: B (67.5 +/- 13.5%) and C (37.9 +/- 11.9%) (p < 0.001). Likewise, for N0 67.5 +/- 13.5%, for N1 47.8 +/- 13.5%, and for N2 9.9 +/- 17% (p < 0.001). CONCLUSIONS: Given the poor results obtained in stages C, particularly stage N2 and also that a better local control is obtained with good tolerance when preoperative radiotherapy is administered, we believe that in order to improve the results it is necessary to initiate preoperative radiotherapy.

[Local control and the esthetic results in the conservative treatment of breast cancer. An analysis of 397 consecutive cases]. / Control local y resultado estético en el tratamiento conservador del cáncer de mama. Análisis de 397 casos consecutivos.

Conservative therapy has been the therapy of choice for patients with breast cancer in early stage. The results of 397 patients treated with conservative therapy and radiotherapy over the breast and lymph node areas (when necessary) are analyzed. The results obtained in the different risk groups and according to the irradiation mode of the tumoral bed are compared. The likelihood of remaining local disease free at 7 years was 94.9 (95% CI: 90.7-99.1). No significant differences were observed regarding the mode of overprinting the tumoral bed: external radiotherapy or brachytherapy, regarding control and aesthetic result; also, no differences were observed between the different risk groups. The existence of a subgroup of patients with contraindication for conservative therapy is currently not demonstrated.

Evidence of genetic heterogeneity in a BALB/c mouse colony as determined by DNA fingerprinting.

A genetic monitoring of the BALB/c mouse foundation colony in our animal facility was carried out. The techniques of choice were skin grafting, coat colour test, flow cytometric analysis for H2 antigens (loci H2-D and H2-A), electrophoretic analysis of isoenzymes (loci Idh1, Pep3, Es3 and Mod1), PCR-amplified microsatellites (loci Igh-V, Ngfg, Plau, Crp, Igh, D16Mit5, D3Mit49 and D17Mit16) and DNA fingerprinting (multilocus probes 33.6, 33.15 and (CAC)5). No evidence of genetic contamination was found, ruling out the possibility of an outcross with AKR, the other albino strain maintained at the facility. Nevertheless, DNA fingerprint patterns revealed evidence of genetic heterogeneity in four out of nine lines of the nucleus colony, interpreted as minisatellite mutations favoured for a single line system with more than 40 generations of separation from the ancestral pair. These mice are mainly used in cancer and immunological research within the institute.

Antigenic analysis of Rhodococcus equi preparations using different horse sera.

An R. equi vaccine, prepared under conditions which induce the expression of many antigens, and which has given encouraging results in field trials, was analyzed by SDS-PAGE and immunoblots and compared with other R. equi preparations: a preparation made in with the same technique from a nonvirulent isolate (virulence associated protein negative, VapA-negative); a whole cell preparation of a VapA-positive R. equi, prepared as a standard bacterin; and a semipurified VapA preparation (APTX). The antigens in these preparations were analyzed using hyperimmune sera (from adult horses vaccinated with the R. equi vaccine), passively and actively immunized foals' sera, asymptomatic but serologically positive foals' sera sera from R. equi pneumonic foals, an equine APTX antiserum, and a VapA monoclonal antibody (Mab). The vaccine under study had many proteins in high concentrations. Hyperimmune sera reacted strongly with vaccine antigens in the high molecular weight regions. In the low molecular weight range, it reacted in the 14 and less kDa zone. Sera from passively immunized foals reacted similarly but not so strongly. Actively immunized foals gave very weak reactions. With the APTX extract, the Mab reacted with bands at 15-17, 44 and 66 kDa; it reacted weakly with the whole cell and not with the VapA-negative preparations. The APTX antiserum and the Mab reacted strongly with the vaccine at the 14 and less kDa zone, and also with bands at 21, 44 and 66 kDa and very tenuously at 18 kDa, but not in the expected 15-17 kDa zone, suggesting that the native form of VapA is altered but without loss of antigenicity in the vaccine preparation. Our results suggest that other higher molecular weight antigens, in addition to VapA, may be important in inducing antibodies that protect young foals from R. equi pneumonia. These antigens are in high concentrations and in an immunogenic form in the vaccine.

[Intervention to improve the quality of continuing drug prescription at a primary care center]. / Intervención de mejora de la calidad de la prescripción farmacológica continuada en un centro de atención primaria.

An evaluation study has been done on the quality of long-term drugs prescription. The aim was to check the effectiveness of certain corrective measures agreed by the professionals working in a primary care centre. Using the results of a first evaluation made in 1989, a series of recommendations were made to reduce the prescription of drugs with a low intrinsic value (LIV). A further evaluation took place 9 months later. A relative reduction of 20.2% in LIV drugs was obtained. This represented a reduction from 25% to 20.8% of the total number of prescriptions. In the relevant pharmacological subgroups a reduction of 77.9% in the prescription of nitrites linked to barbiturites was obtained; of 60% in LIV antacids; of 50.3% in antivaricose drugs; of 36.1% in LIV laxatives; and of 30.1% in Dipiridamol. There was no improvement in the prescription of external LIVs nor cerebral vasodilators. It is concluded that the intervention has proved its usefulness in the improvement of long-term drugs prescription.

[Immune response against foot-and-mouth disease virus in cattle: effect of vaccination]. / Respuesta inmune al virus de la fiebre aftosa en el bovino: efecto de la vacunación.

Foot and Mouth Disease Virus (FMDV) is one of the most feared animal virus and vaccination still has to be used in many countries. In previous reports, using a murine model, we studied the cellular basis of immune responses against FMDV and were able to show that they are atypical. In cattle, although complete protection may be attained after only one dose of killed virus vaccine, very little is known about protection against FMDV, except for antibody responses, but practically nothing concerning the cellular basis of their immune response. Moreover, since neutralizing titers do not always correlate with protection, the potency of vaccines in controlled by viral challenge. Our aim is to study cellular immune responses against FMDV, and to search for a correlate to protection. As a first step, 55 virgin cattle from a non endemic area (Patagonia) were divided into three groups: C: non immunized controls; HS: immunized with saponine containing vaccine; and EO: with oil emulsified vaccine. After vaccination, they were carried to an endemic area (Buenos Aires), where they were challenged with live FMDV. Animals were bled immediately before and 7 days after challenge, and their white blood cells and lymphocyte subpopulations were counted. All animals showed a marked neutropenia and eosinophilia, significantly higher in HS than in EO and C groups; both parameters were significantly better in the 2nd assay. Total lymphocyte counts were normal. Lymphocyte subpopulations were assessed by immunofluorescence using monoclonal antibodies: their proportions were normal and did not change during illness in group C. Several factors could have induced the observed eosinophilia and neutropenia: parasites, stress, saponine, others.(ABSTRACT TRUNCATED AT 250 WORDS)

Respuesta inmune al virus de la fiebre aftosa en el bovino: efecto de la vacunación. / [Immune response against foot-and-mouth disease virus in cattle: effect of vaccination]

Foot and Mouth Disease Virus (FMDV) is one of the most feared animal virus and vaccination still has to be used in many countries. In previous reports, using a murine model, we studied the cellular basis of immune responses against FMDV and were able to show that they are atypical. In cattle, although complete protection may be attained after only one dose of killed virus vaccine, very little is known about protection against FMDV, except for antibody responses, but practically nothing concerning the cellular basis of their immune response. Moreover, since neutralizing titers do not always correlate with protection, the potency of vaccines in controlled by viral challenge. Our aim is to study cellular immune responses against FMDV, and to search for a correlate to protection. As a first step, 55 virgin cattle from a non endemic area (Patagonia) were divided into three groups: C: non immunized controls; HS: immunized with saponine containing vaccine; and EO: with oil emulsified vaccine. After vaccination, they were carried to an endemic area (Buenos Aires), where they were challenged with live FMDV. Animals were bled immediately before and 7 days after challenge, and their white blood cells and lymphocyte subpopulations were counted. All animals showed a marked neutropenia and eosinophilia, significantly higher in HS than in EO and C groups; both parameters were significantly better in the 2nd assay. Total lymphocyte counts were normal. Lymphocyte subpopulations were assessed by immunofluorescence using monoclonal antibodies: their proportions were normal and did not change during illness in group C. Several factors could have induced the observed eosinophilia and neutropenia: parasites, stress, saponine, others.(ABSTRACT TRUNCATED AT 250 WORDS)