Multitarget and suspect-screening of antimicrobials in vegetables samples: Uptake experiments and identification of transformation products.

This work provided an accurate analytical method to perform a multitarget analysis of a variety of antimicrobials (AMs) including sulfonamides, tetracyclines, macrolides, fluoroquinolones and quinolones, one imidazole and one nitroimidazole, one triazole, one diaminopyridine and one derivative of Penicillium stoloniferum in vegetables. The analysis is performed using liquid-chromatography coupled to a low-resolution triple quadrupole mass spectrometer (UHPLC-MS/MS) to detect the target analytesor coupled to a high-resolution q-Orbitrap (HRMS) to monitor the formed transformation products (TPs). Both instruments were compared in terms of limits of quantification and matrix effect at the detection. The method was applied to determine the presence of AMs in organic and non-organic vegetables, where sulfadiazine and mycophenolic acid were detected. On the other hand, the transference of four AMs (trimethoprim, sulfamethazine, enrofloxacin, and chlortetracycline) from soils to lettuces was evaluated through controlled uptake experiments. The choice of AMs was based on the classification into different families, and on the fact that those AM families are the most frequently detected in the environment. In this case, each of the AMs with which the soils were contaminated were found in the exposed lettuces. Moreover, in both studies, specific TPs of the AMs were identified, posing the necessity of assessing their effects in relation to food and human safety.

Comparison of conventional and dispersive solid phase extraction clean-up approaches for the simultaneous analysis of tetracyclines and sulfonamides in a variety of fresh vegetables.

The extensive use of antibiotics in agriculture has led to the occurrence of residual drugs in different vegetables frequently consumed by humans. This could pose a potential threat to human health, not only because of the possible effects after ingestion but also because the transmission of antibiotic-resistant genes could occur. In this work, two accurate sample preparation procedures were developed and validated for the simultaneous analysis of sulfonamides (SAs) and tetracyclines (TCs) in four of the most widely consumed vegetables (lettuce, onion, tomato, and carrot) in Europe. The evaluated protocols were based on QuECHERS for extraction and subsequent clean-up by SPE (solid phase extraction) or dispersive SPE. Parameters affecting both extraction and clean-up were carefully evaluated and selected for accuracy of results and minimal matrix effect. Overall, apparent recoveries were above 70% for most of the target analytes with both analytical procedures, and adequate precision (RSD<30%) was obtained for all the matrices. The procedural limits of quantification (LOQPRO) values for SPE clean-up remained below 4.4 µg kg-1 for TCs in all vegetables except for chlortetracycline (CTC) in lettuce (11.3 µg kg-1) and 3.0 µg kg-1 for SAs, with the exception of sulfadiazine (SDZ) in onion (3.9 µg kg-1) and sulfathiazole (STZ) in carrot (5.0 µg kg-1). Lower LOQPRO values (0.1-3.7 µg kg-1) were obtained, in general, when dSPE clean-up was employed. Both methods were applied to twenty-five market vegetable samples from ecological and conventional agriculture and only sulfamethazine (SMZ) and sulfapyridine (SPD) were detected in lettuce at 1.2 µg kg-1 and 0.5 µg kg-1, respectively.

Static Cold Storage vs Ex Vivo Machine Perfusion: Results From a Comparative Study on Renal Transplant Outcome in a Controlled Donation After Circulatory Death Program.

INTRODUCTION: We aimed to evaluate if ex vivo machine perfusion could minimize the negative impact of cold ischemia on those renal grafts obtained from controlled donation after circulatory death (cDCD). MATERIAL AND METHODS: Prospective observational paired study of kidney transplants from cDCD performed in our center. The kidney from each pair preserved on ice was transplanted first within the first few hours following procurement, while the contralateral kidney was machine-perfused with a LifePort device (Organ Recovery Systems, Brussels, Belgium) and transplanted the following day. RESULTS: A total of 12 cDCDs were included. No differences were observed in delayed graft dysfunction or graft survival between the 2 groups. CONCLUSION: The use of ex vivo perfusion devices is simple and they do not require any large infrastructural or high economic investments, considering the fact that it allows a better selection of recipients and viable organs no longer need to be discarded because of prolonged warm ischemia times.

Effect of Cold Ischemia Time on Kidney Graft Function and Survival: Differences Between Paired Kidney Transplants From the Same Donor.

INTRODUCTION: Kidney transplantation procedures commonly result in a cold ischemia time (CIT) gap when both kidney grafts are implanted in the same center. Owing to logistics, the procedure is usually consecutive, first accomplishing one surgery and then the other. CIT constitutes an independent risk factor for the development of delayed graft function (DGF) in kidney transplants. The effect that CIT exerts on graft and patient survival is still unclear. This study evaluates the relation of CIT and transplant outcomes by comparing paired kidney transplants in terms of survival and graft function. METHODS: We accomplished a retrospective analysis of 402 kidney transplants performed in our center between 2000 and 2017. We selected all transplants where both organs from the same donor were implanted at our hospital, establishing 2 study groups (group 1: first graft implanted and group 2: second graft implanted) to compare by paired data statistical methods. RESULTS: We found an increase in the incidence of DGF in group 2 (42% vs 28.8%; P < .05). Group 2 had significantly worse graft function on day 5 posttransplant (4.7 ± 2.88 vs 3.86 ± 2.8 mg/dL of serum creatinine; P < .05). No significant differences in graft function were found on days 30 and 90 posttransplant. We didn't find any difference in graft survival between both groups. Length of hospitalization stay (17.6 days [± 13] vs 21.6 days [± 17]) and hemodialysis sessions (mean of 2.8 [± 2] vs 3.6 [± 2.2]) were higher in group 2. CONCLUSION: CIT acts as an independent risk factor for the development of DGF in kidney transplantation. CIT had no isolated effect on graft survival.

Usefulness of Delayed Introduction of Tacrolimus in Kidney Transplants Using Type-III Donors After Circulatory Death.

INTRODUCTION: Our study compares 2 immunosuppressive strategies to reduce tacrolimus nephrotoxicity and its risk of acute tubular necrosis: delayed introduction of tacrolimus plus thymoglobulin vs initial tacrolimus plus basiliximab on the results of kidney transplant (KT) using type-III donation after circulatory death (III-DCD). MATERIAL AND METHODS: We analyzed all the transplants performed using type-III DCD in our hospital (42 cases). They were distributed in a first stage with delayed tacrolimus (3°-4° day) + thymoglobulin and a second one with initial tacrolimus + basiliximab, with a follow-up of 6 months. The rate of delayed graft function, the evolution of renal function, and the incidence of rejection were compared. RESULTS: 28 patients received thymoglobulin with delayed tacrolimus, and 13 patients received basiliximab and tacrolimus from day 0 (1 excluded). There were no significant differences in delayed graft function (27% group 1 and 23% group 2) or in rejection (10.7% and 15.4%), respectively. Serum creatinine at day 3, 7, 14, 30, and 180 showed no statistically significant differences. The levels of tacrolimus measured at 10, 30, 90, and 180 days after transplantation were similar, except for the first month: 10.10 ± 2.3 in group 1 and 12 ± 1.7 ng/mL in group 2 (P = .007). CONCLUSIONS: Delayed introduction of tacrolimus does not seem to suppose a benefit in KT using type-III DCD; therefore, the use of thymoglobulin, with its higher profile of adverse effects, seems unjustified in patients with normal immunological risk.

Variability of the Mammalian Target of Rapamycin Inhibitors Is Correlated With Long-Term Renal Graft Survival.

BACKGROUND: Many studies demonstrate the relationship between the high intrapatient variability of calcineurin inhibitor (CNI) levels and poor long-term renal graft outcome. Our objective is to analyze the intrapatient variability observed in the mammalian target of rapamycin inhibitors (mTOR-i) blood levels, to compare the variability of sirolimus (SRL) with that of everolimus (EVL) in kidney transplant patients converted to an mTOR-i, and to analyze whether the coefficient of variation (CV) was correlated with long-term graft survival. METHODS: We analyzed 279 adult renal transplant patients converted to an mTOR-i. CV was calculated using at least 3 blood trough levels between 3 and 18 months postconversion. RESULTS: The mean and median CV of the entire group was 25.54% and 23.7%, respectively. SRL and EVL mean CV was 23.8% and 27.1% (P = .03), respectively. The group of patients into the last tertile with CV> 28.52% presented a lower death-censored graft survival (75.26% vs. 93.01%, P < .0001) with a mean follow-up of 66.5 months. CONCLUSION: The CV of mTOR-i is correlated with long-term renal graft survival, so it should be considered a prognostic factor. SRL has a lower CV than EVL in renal transplant patients converted to mTOR-i in the stable posttransplant phase.

Influence of Proportion of Normal Saline Administered in the Perioperative Period of Renal Transplantation on Kalemia Levels.

BACKGROUND: The hyperchloremic metabolic acidosis triggered by the infusion of normal saline (NS) significantly increases the level of extracellular potassium. In this study we assessed the influence of proportion of NS administered in the perioperative period of renal transplantation on potassium levels in usual clinical practice. METHODS: This study was a retrospective cohort analysis of patients undergoing renal transplantation during a 24-month period (2015-2016). To determine the influence of NS on K+ levels, simple linear regression and multiple linear regression analyses were performed, adjusted for the total volume of fluids administered, establishing the difference in serum K+ levels for each 20% increase in the proportion of NS. RESULTS: As the proportion of NS administered increased, K+ levels at 24 hours were significantly increased (P = .026) (0.69 mEq/L K+ increase per 20% increase in NS ratio). Mean K+ values at 24 hours (adjusted for total volume of fluids administered) ranged from 4.17 mEq/L (95% confidence interval [CI] 3.7-4.56) in patients who did not receive NS to 4.85 mEq/L (95% CI 4.56-5.15) in those administered exclusively NS. CONCLUSION: The risk of developing hyperkalemia in patients who receive a balanced solution with potassium in its formulation compared with NS in the perioperative period of renal transplantation is not increased, but the volume of NS administered is significantly associated with increases in K+ levels at 24 hours.

Improving the Outcomes of Organs Obtained From Controlled Donation After Circulatory Death Donors Using Abdominal Normothermic Regional Perfusion.

The use of donation after circulatory death (DCD) has increased significantly during the past decade. However, warm ischemia results in a greater risk for transplantation. Indeed, controlled DCD (cDCD) was associated with inferior outcomes compared with donation after brain death. The use of abdominal normothermic regional perfusion (nRP) to restore blood flow before organ recovery in cDCD has been proposed as better than rapid recovery to reverse the effect of ischemia and improve recipients' outcome. Here, the first Spanish series using abdominal nRP as an in situ conditioning method is reported. A specific methodology to avoid restoring circulation to the brain after death determination is described. Twenty-seven cDCD donors underwent abdominal nRP during at least 60 min. Thirty-seven kidneys, 11 livers, six bilateral lungs, and one pancreas were transplanted. The 1-year death-censored kidney survival was 91%, and delayed graft function rate was 27%. The 1-year liver survival rate was 90.1% with no cases of ischemic cholangiopathy. Transplanted lungs and pancreas exhibited primary function. The use of nRP may represent an advance to increase the number and quality of grafts in cDCD. Poor results in cDCD livers could be reversed with nRP. Concerns about restoring brain circulation after death are easily solved.

Relationship Between Albuminuria During the First Year and Antibody-Mediated Rejection in Protocol Biopsies in Kidney Transplant Recipients.

BACKGROUND: Antibody-mediated rejection is the main cause of deterioration of kidney transplants and frequently is detected only by means of protocol biopsies. The aim of this study was to relate the presence of albuminuria throughout the 1st year to the histologic findings detected by 1-year protocol biopsies in kidney graft recipients. METHODS: Retrospective observational study of 86 protocol biopsies 1 year after transplantation. Albuminuria was measured at 3, 6, 9, and 12 months in urine samples and expressed as albumin/creatinine (mg/g). RESULTS: Analysis of biopsies, reflected according to the Banff criteria, the following categories: fibrosis and tubular atrophy, 35 (40.7%); cellular rejection, 13 (15.1%); antibody-mediated rejection, 8 (9.3%); chronic glomerulopathy, 10 (11.6%); normal, 14 (16.3%); recurrence, 1 (1.2%); and other, 5 (5.8%). The proportions of patients with albuminuria for Banff scale scores (0 vs ≥1, respectively) at 6 and 12 months, respectively, after transplantation, were: for marker glomerulitis, 45.5% versus 59.3% (P = .021) and 36.4% versus 70.4% (P < .001); for marker glomerulopathy, 49.1% versus 50.0% (P = .051) and 42.1% versus 58.3% (P = .019); for marker peritubular capillaritis, 45.8% versus 60.9% (P = .047) and 39.0% versus 69.6% (P = .276); and for marker C4d, 49.2% versus 56.3% (P = .894) and 46.2% versus 56.3% (P = .774). CONCLUSIONS: The presence of albuminuria after renal transplantation is common, especially in patients with proteinuria. Persistent albuminuria after transplantation, even at low levels, can be indicative of subclinical antibody-mediated rejection. Additional broader studies to relate the albuminuria to histologic changes observed in protocol biopsies are required.

[Changes in the pre-transplant bone-mineral metabolism do not affect the initial progress of the renal graft]. / Las alteraciones del metabolismo óseo-mineral pretrasplante no influyen en la evolución inicial del injerto renal.

BACKGROUND: Abnormalities in serum calcium, phosphate, and Parathyroid Hormone (PTH) concentrations are common in patients with chronic kidney disease and have been associated with increased morbidity and mortality. One of the most common problems in the first weeks after renal transplantation is Delayed Graft Function (DGF). There are several well-known risk factors for DGF development, but the role of calcium phosphate-PTH homeostasis as a risk factor for early graft dysfunction is controversial. This issue was addressed in the current study. METHODS: Pretransplant PTH, calcium and phosphate values were gathered in 449 patients that received a renal transplant in our center between 1994 and 2007. Other variables expected to influence the risk for delayed graft function were included from the clinical charts. RESULTS: The incidence of DGF was 27.3%. DGF development was significantly associated with recipient age, type and need of renal replacement therapy, peak panel reactive antibodies, transfusion number and donor age. There were no significant differences in the mean pretransplant values of calcium (9.4 +/- 1.0 vs. 9.5 +/- 0.9 mg/dl, p = 0.667), phosphate (5.7 +/- 1.8 vs. 5.5 +/- 1.5 mg/dl, p = 0.457), calcium-phosphate product (53.5 +/- 17.2 vs. 51.8 +/- 14.6 mg(2)/dl(2), p = 0.413) and PTH (315 +/- 312 vs. 340 +/- 350 pg/ml, p = 0.530) between patients with and without DGF. CONCLUSIONS: In our study population pretransplant serum PTH, calcium and phosphorus levels have no influence on the risk for DGF.

Las alteraciones del metabolismo óseo-mineral pretrasplante no influyen en la evolución inicial del injerto renal / No disponible

Antecedentes: el Retraso en la Función del Injerto (RFI) es uno delos problemas más frecuentes en las primeras semanas del trasplante renal, afectando a su evolución. Conocer los factores de riesgo de RFI puede ayudar a reducir su incidencia. Las alteraciones en los niveles séricos de calcio, fósforo y Hormona Paratiroidea (HPT) son muy frecuentes en los pacientes en lista de espera de trasplante y podrían favorecer la aparición de RFI. Sin embargo, diversos estudios que han analizado la relación entre los niveles pretrasplante de calcio, fósforo y HPT y el desarrollo de RFI han obtenido resultados dispares que no permiten confirmar ni descartar que influyan en el mismo. Métodos: estudiamos los valores pretrasplante de calcio, fósforo y HPT en 449 pacientes trasplantados renales realizados entre 1994 y 2007. Se definió RFI en aquellos pacientes que precisaron diálisis durante la primera semana postrasplante. De las historias clínicas se recogieron los datos clínicos y analíticos relacionados con RFI. Resultados: un 27,3%presentó RFI. Los factores significativos de riesgo para desarrollar RFI fueron la edad del receptor, el tipo y la necesidad de tratamiento sustitutivo renal, el título de anticuerpos anti-HLA máximos, el número de trasfusiones pretrasplante y la edad del donante. No detectamos diferencias significativas en los valores medios de calcio (9,4 ± 1,0 vs. 9,5 ± 0,9 mg/dl, p = 0,667), fósforo(5,7 ± 1,8 vs. 5,5 ± 1,5 mg/dl, p = 0,457), producto fosfocálcico (53,5± 17,2 vs. 51,8 ± 14,6 mg2/dl2, p = 0,413) y HPTi (315 ± 312 vs. 340± 350 pg/ml, p = 0,530) en los pacientes con y sin RFI. Conclusiones: en nuestro estudio, los parámetros séricos pretrasplante del metabolismo óseo-mineral no favorecen el desarrollo de RFI (AU)

Background: abnormalities in serum calcium, phosphate, and Parathyroid Hormone (HPT) concentrations are common in patients with chronic kidney disease and have been associated with increased morbidity and mortality. One of the most common problems in the first weeks after renal transplantation is Delayed Graft Function (DGF). There are several well-known risk factors for DGF development, but the role of calciumphosphate-HPT homeostasis as a risk factor for early graft dysfunction is controversial. This issue was addressed in the current study. Methods: Pretransplant HPT, calcium and phosphate values were gathered in 449patients that received a renal transplant in our center between 1994 and 2007. Other variables expected to influence the risk for delayed graft function wereincluded from the clinical charts. Results: The incidence of DGF was 27.3%. DGF development was significantly associated with recipient age, type and need of renal replacement therapy, peak panel reactive antibodies, transfusion number and donor age. There were no significant differences in the mean pretransplant values of calcium (9.4 ± 1.0 vs. 9.5 ± 0.9 mg/dl, p = 0.667),phosphate (5.7 ± 1.8 vs. 5.5 ± 1.5 mg/dl, p = 0.457),calcium-phosphate product (53.5 ± 17.2 vs. 51.8 ± 14.6mg2/dl2, p = 0.413) and HPT (315 ± 312 vs. 340 ± 350pg/ml, p = 0.530) between patients with and without DGF. Conclusions: In our study population pretransplant serum HPT, calcium and phosphorus levels have no influence on the risk for DGF (AU)

Evaluation of the efficacy and safety of the conversion from a calcineurin inhibitor to an everolimus-based therapy in maintenance renal transplant patients.

Everolimus has recently been introduced into clinical practice with promising perspectives due to its efficacy, lack of nephrotoxicity, and antitumor effects. Experience in clinical trials associated with low-dose cyclosporine showed good results, but there is almost no experience in calcineurin inhibitor (CNI) elimination learning it as the primary immunosuppressant. We describe our experience in a series of 78 stable renal transplant patients who were switched to Everolimus with complete and quick elimination of the CNI: the procedure of conversion, pharmacokinetic results after conversion, evolution of renal parameters (renal function, proteinuria, and others), and safety data (acute rejection and adverse events). An initial dose of 3 mg/d was adequate to obtain the recommended trough levels between 5 and 10 ng/mL. Our results demonstrated that conversion to Everolimus was a simple, safe procedure that must be considered in patients CNI toxicity, especially those with malignant neoplasms and progressive deterioration of renal function due to chronic allograft nephropathy.

Elemental distributions in femoral bone of rat under osteoporosis preventive treatments.

One of the abnormalities of bone architecture is osteoporosis as occurring in post-menopausal women. Especially long bones, such as femur, become more fragile and more prone to fracture. The efficiency of several osteoporosis preventative treatments based on oestrogen and progestin in bone structure and mineral recovery was studied using ovariectomized Wistar rats as an osteoporosis experimental model. Diagonal cross-sections of the proximal epiphysis of femoral bones were analysed using nuclear microscopy techniques in order to map and determine the concentration profiles of P, Ca, S, Fe and Zn from the epiphysis to diaphysis and across the cortical and trabecular bone structures. In control animals (not ovariectomized), the S and Zn contents significantly characterized differences between cortical and trabecular bone structures, whereas P and Ca showed increased gradients from the epiphyseal region to the diaphysis. After ovariectomy the differences observed were differential according to the type of hormonal supplementation. A significant decrease in P and Ca contents and depletion of minor and trace minerals, such as S, Fe and Zn, were found for both cortical and trabecular bone structures after ovariectomy relative to controls. Bone mineral contents were reversed to control levels by synthetic oestrogen supplementation, and combined oestrogen and progesterone treatment. Recovery was more evident in the femoral epiphysis and neck than in the diaphysis. The use of oestrogen alone did not lead to bone recovery after ovariectomy. Alterations in bone mineral composition observed for animals receiving synthetic oestrogen and combined oestrogen and progesterone supplement might reflect beneficial structural changes in critical regions of long bones, mostly affected in post-menopausal osteoporosis.

[Microscopic polyangiitis in a patient with rheumatoid arthritis]. / Poliangeítis microscópica en paciente con artritis reumatoide.

Rheumatoid arthritis (RA) is a systemic disorder that primary involves joints, although renal disease has also been associated it is not common that rapidly progressive glomerulonephritis (RPGN) appears. We report the case of a patient with nodular and aggressive RA who had an acut renal failure secondary to ANCA positive RPGN due to a Microscopic polyangiitis who was not responsive to steroids and cyclophosphamide therapy.

Effect of hormone replacement therapy on the elemental contents of uterine tissue.

For the past years, different therapies based on steroid hormone supplementation or modulators of estrogen receptors have been used after menopause to prevent or manage osteoporosis. Although these treatments seem to be beneficial, they have some negative effects in the uterus and breast. The objective of this study was to assess variations for the concentrations of K, Ca, Mn, Fe, Cu, Zn, and Se in uterine tissue of Wistar rats. Ovariectomized rats were subjected to estrogen, progesterone, raloxifene, and tibolone supplementation and compared with nonovariectomized control animals. Elemental contents determined by the particle-induced X-ray emission (PIXE) technique revealed major alterations in Fe, Ca, Mn, and Se in the uterus of ovariectomized rats relative to control animals. After ovariectomy, a significant increase in Ca and Fe and a significant decrease in Mn and Se contents were determined in the uterus. For the ovariectomized groups in which animals received raloxifene, tibolone, estrogen, and estrogen combined with progesterone supplementation, an overall recovery in Mn, Fe, and Se contents was verified. Elemental concentration in the progesterone-supplemented group did not significantly differ from ovariectomized animals receiving placebo. The alterations found for ovariectomized animals receiving placebo and progesterone suggest tissue impairment and trace element imbalance, contrasting with the remaining supplemented groups where an enhancement of tissue activity might justify similar concentration levels relative to controls, because most of the elemental contents altered after ovariectomy.

Immunoprophylaxis with Simulect (basiliximab) in kidney transplantation: results from routine clinical practice at 18 kidney transplant units.

OBJECTIVE: The objective of this study is to assess a Simulect (basiliximab) regimen in routine clinical practice in the Spanish kidney transplantation units to evaluate efficacy and safety. METHODS: In this prospective, observational study, data on demographics, parameters of efficacy, and safety in patients who under with kidney transplantation treated with Simulect (basiliximab) were collected through an on-line collection system. RESULTS: One hundred sixty three patients at 18 kidney transplant units included 12 months follow-up. The patient mean age was 52 years (DS 13,67) including 96 (58.90%) men and 67 (41.10%) women. Cold ischemia time was 19 hours (DS 6,79). Only 2 patients presented with PRA >50%. For prophylactic immunosuppression, 67.13% of patients received triple therapy with CNI (cyclosporine 49.65% or tacrolimus 17.48%), MMF (66.43%) or AZA (10.49%), and steroids. Incidence of acute rejection (AR) at 12 months was 12.27% (1.84% steroid-resistant). In subgroup analysis, AR was 13.5% in nondiabetics and 4.5% in diabetics, including 3 steroid-resistant episodes (1.84%) in nondiabetics and none in diabetics. In relation to donor age, AR was incidence 10.3% in patients with kidneys from donors aged 50 years or younger and 10.6% when donors were older than 50 years, including 1 (1.73%) and 2 (1.93%) steroid-resistant episodes, respectively. The graft and patient survival rates at 12 months were 90% and 98%, respectively. CONCLUSIONS: Simulect (basiliximab) used in routine clinical practice provided good prophylaxis against acute rejection in several kidney transplant patient populations, similar to that observed in randomized clinical studies with excellent tolerability and safety.

Influence of estradiol and gestagens on oxidative stress in the rat uterus.

OBJECTIVE: We studied the effect of ovariectomy, estradiol (E2), and E2 + medroxyprogesterone (MPA) on the Wistar rat uterus. METHODS: We used 15 adult female rats. The study was divided into the following four stages: (a) extirpation of the upper half of the left hemi-uterus (basal state) and ovariectomy; (b) animals were maintained for 15 days without treatment, performance of a new laparotomy, and extirpation of the remaining left hemi-uterus (OVX state); (c) beginning of E2 replacement therapy (ERT) (8 microg/day) for 15 days, followed by extirpation of the upper half of the right hemi-uterus (ERT state); and (d) the administration of E2 was continued, and oral treatment with MPA was begun (20 microg/day) to last for a further 15 days. At the end of the combined hormone replacement therapy (HRT) the remaining right hemi-uterus was extirpated (HRT state). At the end of each intervention, the plasma concentrations of E2 and PRG were measured. RESULTS AND DISCUSSION: The ovariectomy significantly reduced the malonaldehyde (MDA) levels (P < 0.0008) and catalase activity (P < 0.0006). The ERT very significantly (P < 0.0033) raised the catalase and MDA levels; these significance levels were maintained after the Bonferroni method was applied (overall error 5%). The HRT reduced the levels of MDA and catalase, but not significantly after the Bonferroni test was applied.Conclusions. Uterine oxidative stress is increased by E2, resulting in a significant increase in MDA. This may be modulated in part by the catalase activity. Although it cannot be confirmed categorically, MPA seems to intervene by decreasing the said oxidative stress.

Influence of estrogen replacement therapy on plasma lipid peroxidation.

OBJECTIVE: To observe whether any relationship exists between the concentration of plasma estradiol (E2) and the plasma concentrations of malondialdehyde (MDA) or whether a relationship exists between the concentration of plasma E2 and the activity of the erythrocyte enzymes, superoxide dismutase (SOD) and catalase, in ovariectomized female Wistar rats (treated and untreated with E2). DESIGN: We used 40 ovariectomized Wistar rats randomly assigned to four groups. The first group was allowed to evolve freely with no treatment. A gel containing 17beta-estradiol was administered transdermally to the other three groups at doses of 4, 8, and 16 microg/day, respectively. After 15 days of treatment, blood samples were obtained from the four groups. The concentrations of plasma MDA and E2 and the activities of erythrocyte catalase and SOD were determined. RESULTS: There were significant correlations between the MDA levels and the logarithm (base 10) of the plasma E2 concentrations in both linear (p = 0.00093) and quadratic (p = 0.000001) regression analyses. No relationship was found between the E2 concentrations and the catalase and SOD activities. CONCLUSIONS: There was a clear relationship between the plasma levels of MDA and the logarithm of the plasma E2 concentrations, which was best demonstrated with a quadratic regression. This model may explain the contradictory findings presented by estrogens with respect to their pro-or antioxidant action.

A phase I clinical trial of a multi-epitope polypeptide TAB9 combined with Montanide ISA 720 adjuvant in non-HIV-1 infected human volunteers.

A phase I clinical trial was performed to examine the safety and immunogenicity of a multi-epitope polypeptide comprising the central 15 amino acids of the V3 loop from six HIV-1 isolates. This protein called TAB9 was emulsified in Montanide ISA720 (Seppic, Paris) and administered intramuscularly at doses of 0, 0.2 and 1 mg to 24 healthy, HIV-1 seronegative adult males. Three immunisations were given at months 0, 1 and 6 in a randomised, double blind, placebo controlled clinical trial. The placebo was generally well tolerated. However, severe local reactions were observed in TAB9 vaccinated subjects after the second and third inoculations. Seven out of eight volunteers from the lower dose group showed moderate or severe local inflammation, while four out of eight subjects from the higher dose group developed granulomas and sterile abscesses. In general, the reactogenicity depended on the number of inoculations given and the dose of TAB9. Both doses were immunogenic, all immunised volunteers seroconverted and antibodies were broadly reactive against the V3 peptides included in the protein. All vaccine's sera reacted against gp120 in Western blot and 50% of them also neutralised at least one out of five laboratory isolates tested. No differences between doses were found. Anti TAB9 lymphoproliferative responses were observed, being more intense in the high dose group. Due to the strong local reactions that were found in this study, a change in the formulation will be required for further trials with this vaccine candidate in humans.

Influence of estradiol on oxidative stress in the castrated rat uterus.

OBJECTIVE: We set out to study how the concentration of estradiol influences oxidative stress (malondialdehyde) and the superoxide dismutase and catalase antioxidant systems in the castrated female Wistar rat uterus. METHODS: We used 28 castrated female Wistar rats: 7 were left to evolve freely and the rest were divided into three groups of 7 animals receiving respective doses of 4, 8, and 16 microg/day of estradiol (E2) for 15 days. At the end of the study period, we determined the plasma concentrations of E2 and the concentrations of malondialdehyde (MDA) and superoxide dismutase and catalase activities in the uterus. RESULTS: There was a significant correlation (P < 0.000028) between the uterine malondialdehyde levels and the logarithm (base 10) of the plasma E2 concentrations and also between malondialdehyde and the uterine catalase activity (P < 0.002). The regression plane that best fitted the correlation among the three variables was MDA = 10.21 + 12.88 x Log [E2] - 0.49 x catalase activity. We found no significant relationships with the superoxide dismutase activity. CONCLUSIONS: There was a linear correlation between the base-10 logarithm of the estradiol plasma concentration and the phenomenon of uterine lipid peroxidation as measured by the MDA concentration in the uterus. This phenomenon was in part modulated by the inverse linear relationship between the antioxidant activity of the uterine catalase and the concentration of uterine MDA.