The effect of selenium supplementation before calving on early-lactation udder health in pastured dairy heifers.

Selenium (Se) deficiency has been associated with lowered resistance to mastitis in dairy cattle. However, little published data exists on the effect of Se supplementation before calving on udder health of pastured dairy heifers. Further, the relative efficacy of injectable barium selenate and oral organic Se for improving udder health in cows has not previously been tested. The objectives of this study were to determine the effects of precalving Se supplementation and type of supplementation on the blood activity of glutathione peroxidase and measures of udder health immediately after calving and during the first month of lactation in pastured dairy heifers. One hundred forty pregnant Chilean Holstein-Friesian heifers were fed a basal diet containing, on average, 0.15 mg of Se/kg of dry matter. One month before predicted calving, heifers were allocated to 1 of 3 groups. Group 1 (n=49) received no supplementary Se, group 2 (n=46) received a single subcutaneous injection of Se (1 mg/kg of live weight, as barium selenate), and group 3 (n=45) was fed Se yeast (3 mg/heifer/d until calving). Heifers supplemented with barium selenate had a higher glutathione peroxidase activity from 14 d in milk onwards. Selenium supplementation, irrespective of source, tended to reduce the prevalence of intramammary infection (IMI) and decrease the prevalence of quarters with high somatic cell count (SCC) at calving. Overall, Se supplementation did not result in a reduction of the incidence of new IMI or clinical mastitis or in decreased SCC during the balance of the first month of lactation. However, in pasture-based heifers injected with barium selenate before calving, and fed diets with 1.3 and 2.5 mg of Se/d precalving and during lactation, respectively, no cases of clinical mastitis were observed in the first month of lactation.

Educational program of organ donation and transplantation at medical school.

A favorable attitude of health professionals to organ donation can positively influence the decision of families of potential donors. By increasing health professionals knowledge about donation and transplantation and qualifying them to disseminate information, education has produced a positive response to increase the insufficient number of donors. Educating students early in their careers may become crucial in this setting. In order to supply the necessary information about the process of donation and transplantation, a medical school in association with the Hospital Transplant Coordination Department created an educational program of organ donation and transplantation. This course is intended for medical, biomedical, and nutrition students. The objective of our program is to supply basic knowledge about organ donation and transplantation to students of medicine, nutrition, and biomedicine and to enhance their commitment to this process. Each semester, 50 to 90 students are enrolled in the course, which involves a total of 25 hours. Various aspects are approached such as brain death, donor management, political and legal aspects of donation, and skin, lung, bone marrow, heart, pancreas, liver, and kidney transplantation. Between March 2006 and June 2007, three courses were carried out and 200 students were trained. The students evaluated the course and rated it as excellent, concluding that it contributed to their education. Their attitude toward organ donation and transplantation was strongly positive at the end of the course. This project aims to educate and stimulate students in the process of organ donation and transplantation and should be implemented in other medical schools.

Clinical relevance of low levels of preformed alloantibodies detected by flow cytometry in the first year post-kidney transplantation.

OBJECTIVE: To determine the prevalence of transplants performed with a false-negative cytotoxicity cross-match and to analyze the clinical relevance of alloantibodies (Ab) detected only by flow cytometry (flow). METHODS: We studied 66 patients undergoing kidney transplantation from a cadaveric donor. All patients had a simultaneous negative T+AHG+DTT and B+DTT. Pretransplant sera were retrospectively analyzed by flow cytometry according to an Emory University protocol: (1) T+ and B-: Ab anti-class I; (2) T- and B+: anti-class II; (3) T+B+: anti-class I + II. Chi-square, Fisher exact, Student t test, and Kaplan Meier analyses were employed with significance assigned at P < or = .05. RESULTS: The overall incidence of false-negative cytotoxicity was 33.3% (22/66), namely, 6.1% (n = 4) anti-class I; 9.1% (n = 6) anti-class II; and 18.2% (n = 12) anti-class I + II. Primary nonfunctioning grafts occurred in 6.8% (3/44) and 13.6% (3/22) negative and positive flow patients (two anti-class I + II and one class II; P = .39). The incidence of graft loss in the first year was respectively, 13.6% (6/44) and 18.2% (4/22; two anti-class II and two anti-class I + II; P = .72). Compared to flow-negative grafts, creatinine levels were significantly higher among flow-positive patients at 8 and 12 weeks. One-year graft survivals were 86.4% among negative versus 81.8% for the positive group (P = .67). CONCLUSIONS: We observed that 33% of kidney transplant recipients had low levels of alloantibodies detected only by flow. This single factor was associated with the worst graft function in the first trimester with a suggestion of a higher risk for non-functioning graft.

Immunoglobulin G-positive in B-cell cross-match decreases kidney allograft survival.

We retrospectively studied all 1149 transplants performed at our center between 1993 and 2003 to determine the incidence and clinical effect of pretransplant B-positive cross-match on kidney graft survival. The patients were divided in two groups: B-negative (n = 1102) and B-positive in current sera (n = 47; 4.1%). AB-positive test was more frequent among regrafted patients (14% vs 3%; P = .00). Demographic data were not different between the groups. The overall rate of graft loss was similar (26% vs 24%, respectively; P = .86). However, early nonsurgical graft losses were more frequent among B-positive patients (46% vs 20%, respectively; P = .04). IgM was the most frequent immunoglobulin in the B-positive group (76% IgM and 24% IgG). There was no significant difference between B-negative and B-positive groups in the 1-, 5-, and 10-year graft survival rates (87% vs 83%, 73% vs 78%, 64% vs 66%, respectively; P = .87). The graft survival was significantly reduced comparing an IgG anti-B cell to the B-negative group (P = .03) as well as IgG compared to IgM (P = .004). In conclusion, only B-positive cross-match due to IgG decreased graft survival. Even though it is an uncommon situation (0.9%), this study stressed the clinical value of the B-cell cross-match as a tool to identify patients with a higher immunological risk.

Antigenemia for cytomegalovirus in renal transplantation: choosing a cutoff for the diagnosis criteria in cytomegalovirus disease.

Cytomegalovirus (CMV) infection is a frequent complication in transplant recipients, causing a high level of morbidity and mortality. We studied 203 consecutive renal transplant recipients performed between January 2000 and December 2001. Patients underwent weekly measurements of CMV pp65 antigen to assess CMV activity from the 4th to the 12th week posttransplantation. The results were reported as number of cells positive for the pp65 antigen among 10(5) granulocytes. In order to define a best cutoff to diagnose CMV disease with desirable sensitivity and specificity, we used a receiver operator characteristics (ROC) curve. The cutoff of four positive cells corresponded to a sensitivity of 93% and specificity of 60% (AUC = 0.87) for the diagnosis of CMV disease. The chosen cutoff for starting antiviral treatment was 10 cells, since this was associated with a sensitivity of 92% and specificity of 70% (AUC = 0.90). In conclusion, the highly sensitive cutoff points for the diagnosis of antigenemia was four cells and 10 cells for initiation of antiviral therapy.

Off-pump coronary artery bypass grafting does not reduce lymphocyte activation.

OBJECTIVE: In this study, we test the hypothesis that off-pump coronary bypass surgery might result in less lymphocyte activation than on-pump coronary surgery. We also study the behavior of lymphocyte activation markers during and after surgery. BACKGROUND: Coronary artery bypass surgery is known to be associated with changes of inflammatory mediators, immune function, and early phase lymphocyte activation, which could cause postoperative lymphopenia and lymphocyte unresponsiveness. METHODS: We studied lymphocyte activation response in 28 patients randomized to off-pump (n = 13) or on-pump (n = 15) coronary artery bypass surgery. Expression of CD25, CD26, CD69, and DR on T (CD3+) and B (CD19+) lymphocytes on peripheral blood was assessed through flow cytometry. RESULTS: The response of T lymphocytes and their activation markers, as well as B lymphocytes and their activation markers, was similar after on- and off-pump surgery. Overall, T lymphocytes decreased to the lowest level 9 h after surgery and tended to increase later. For B lymphocytes, there was early reduction with increase on the 1st postoperative day. There was early activation of CD69+ and late activation of CD25+ on T lymphocytes. For B lymphocytes, there was early activation of CD69+ and late activation of DR+. CONCLUSIONS: (1) Compared to on-pump cardiopulmonary bypass, off-pump surgery does not reduce lymphocyte activation. (2) Coronary bypass surgery causes the early activation of lymphocytes, as evidenced by the increased expression of lymphocyte activation markers.

Expanding criteria for the use of living donors: what are the limits?

The cadaver organ shortage has pushed the transplant community to extend the boundaries beyond the traditional criteria used for living donor transplantation. This new liberal policy involves: (1) the type of donor, such as emotionally related individuals, the direct or indirect interchange of donors, anonymous as well as rewarded donation; (2) challenging immunological criteria, using incompatible ABO blood types and or transplantation across a positive cross-match; (3) relaxing clinical criteria related to elderly, hypertensive, or obese donors, or patients with nephrolithiasis, fibromuscular renal artery disease, hematuria, or renal cell carcinomas. However, these practices may be dangerous. They must be clearly validated to promote a liberal policy of donor acceptance since it may carry a risk for both the donor and the recipient as well as for society. It is crucial to ensure the physical integrity of the donor as well as to provide guarantees, for instance a 1-year policy of life insurance, an indefinite long-term medical follow-up and the assurance of going to the top of the waiting list if the donor becomes uremic in the future.

Comparison between RFLP-PCR and antigenemia for pp65 antigen for diagnosis of cytomegalovirus disease after kidney transplantation.

Cytomegalovirus (CMV) infection is the single most frequent infectious complication in renal transplant recipients. The purpose of this study was to analyze the diagnostic efficacy of PCR-RFLP compared to antigenemia for CMV disease (CMVD) in kidney transplant recipients. From November 2001 to February 2002, 19 renal adult transplant recipients were followed with weekly measurements of CMV pp65 antigen to monitor the activity of CMV from the week 4 to 12 posttransplantation. Only 4 (21.1%) patients did not develop viremia during the first 12 posttransplantation weeks. Active infection was observed in 15 patients (78.9%): asymptomatic viremia in 6 (31.6%) and CMVD in 9 (47%). All patients who developed CMVD showed positivity in both methods during the observation period. The number of positive cells ranged from 11 to 292 cells in patients with CMVD and one to eight cells among those with asymptomatic viremia. Both methods revealed 100% sensitivity for CMVD diagnosis. The specificity was 60% for antigenemia and 70% for PCR, with positive predictive values of 60% and 75%, respectively.

Cytomegalovirus disease latent and active infection rates during the first trimester after kidney transplantation.

Cytomegalovirus (CMV) infection is the single most frequent infectious complication in renal transplant recipients. The aim of this study was to determine the incidence of latent and active infections with CMV during the first 3 months after kidney transplantation. From January 2000 to December 2001, 203 consecutive adult renal transplant recipients underwent weekly measurements of pp65 CMV antigen from the 4th to the 12th posttransplantation week. Latent infection (seropositivity) was found in 92% of the population. Primary infection occurred in 4.9% (10 of 203), among whom 66% were previously seronegative patients. Among the primary infection patients, 70% (7 of 10) developed severe disease. The overall incidence of viremia was 69.5%, being more frequent among cadaver recipients (79% vs 59%; P =.02). The overall incidence of CMV disease was 38.4% (78 of 203) with 24.6% classified as severe disease requiring antiviral therapy. In conclusion, our population showed a high prevalence of latent infection with viremia. Not all patients developed clinical disease. Most subjects experienced a mild spectrum of symptoms, probably due to the prospective search for active infection during the major risk period after kidney transplantation.

Prevalence of TEL/AML1 fusion gene in Brazilian pediatric patients with acute lymphoblastic leukemia.

We studied 58 childhood B-lineage acute lymphoblastic leukemia (B-ALL) in Brazilian sample patients at the time of diagnosis to investigate the prevalence of the cryptic t(12;21)(p13;q22). All bone marrow specimens were G-band karyotyped, and commercial dual-color DNA probes were used to search for fusion signals in nuclei. The karyotype analysis showed hyperdiploidy as the most frequent abnormality. The frequency of patients with TEL/AML1 gene fusion was 19% (11 out of 58 cases). Six of the positive samples had normal karyotypes. Deletion of the wild-type TEL allele was observed in 27.3% of TEL/AML1 fusion-positive cases, but it was also identified in 4.2% of the negative cases. Three cases presented two fusion signals, indicating possible duplication of the der(21). The mean age of the patients with TEL/AML1 fusion was 4.8 years and the mean amount of peripheral leukocytes was 44,270 x 10(6)/L. The higher frequency of females with B-ALL (33/58 cases) observed in our sample was probably due to the selection mode of the study cases. The prevalence of TEL/AML1 fusion in Brazilian children in our study is similar to that found in other populations.

Lymphocyte subpopulations during cytomegalovirus disease in renal transplant recipients.

We have determined the number of circulating T, B and natural killer cells in renal transplant recipients in order to detect changes during cytomegalovirus (CMV) infections. Serial blood samples were taken from 61 patients on standard triple immunosuppression therapy (cyclosporin A, azathioprine and prednisone). Using two-color flow cytometry analysis, the absolute number of CD3+, CD4+, CD8+, CD19+, CD3+HLA-DR+ and CD16+56+ cells was determined. Forty-eight patients (78.7%) developed active CMV infection, and all of them subsequently recovered. Twenty of the infected patients (32.8%) presented symptoms compatible with CMV disease during the infectious process. The number of lymphocytes and their main subpopulations were normal before the onset of CMV disease. During the disease there was a decrease followed by a significant increase (P<0.005) in the number of CD3+, CD4+, CD8+ and CD3+HLA-DR+ cells. No significant changes were observed in natural killer cells or B lymphocytes during the disease. We conclude, as observed in all viremic patients recovering from infection, that recovery is associated with an increase in the number of T cell subsets. The monitoring of different lymphocyte subsets along with antigenemia can be extremely useful in the detection of patients at high risk of developing CMV symptoms, allowing the early introduction of antiviral therapy or the reduction of immunosuppression therapy.

Lymphocyte subpopulations during cytomegalovirus disease in renal transplant recipients

We have determined the number of circulating T, B and natural killer cells in renal transplant recipients in order to detect changes during cytomegalovirus (CMV) infections. Serial blood samples were taken from 61 patients on standard triple immunosuppression therapy (cyclosporin A, azathioprine and prednisone). Using two-color flow cytometry analysis, the absolute number of CD3+, CD4+, CD8+, CD19+, CD3+HLA-DR+ and CD16+56+ cells was determined. Forty-eight patients (78.7 percent) developed active CMV infection, and all of them subsequently recovered. Twenty of the infected patients (32.8 percent) presented symptoms compatible with CMV disease during the infectious process. The number of lymphocytes and their main subpopulations were normal before the onset of CMV disease. During the disease there was a decrease followed by a significant increase (P<0.005) in the number of CD3+, CD4+, CD8+ and CD3+HLA-DR+ cells. No significant changes were observed in natural killer cells or B lymphocytes during the disease. We conclude, as observed in all viremic patients recovering from infection, that recovery is associated with an increase in the number of T cell subsets. The monitoring of different lymphocyte subsets along with antigenemia can be extremely useful in the detection of patients at high risk of developing CMV symptoms, allowing the early introduction of antiviral therapy or the reduction of immunosuppression therapy

Tuberculosis infantil en Valdivia: Experiencia clínica en 16 años / Tuberculosis in infancy: sixteen year experience

A clinical experience of tuberculosis (TB) in 104 pediatric patients observed at the Hospital Regional de Valdivia (southern Chile), along a sixteen year period (1986 - 2001) is presented. Pulmonary TB was diagnosed in 88 cases, extrapulmonary TB in 19 cases and both localizations in three cases. Pulmonary TB was bacteriologically confirmed in 62.5% of cases, through either gastric aspirate or sputum samples. Gastric aspirate yielded 42.8% of positive cultures; two infants who presented central nervous system involvement, died. A case of congenital TB was observed in a 30 day old infant. There were no cases of AIDS in this experience. Pediatric TB has progressively decreased its incidence in our region, particularly the extrapulmonary localization, but still represents a challenging disease to clinicians.