Uterus transplantation in a sheep model: novel surgical technique with long-term survival and uterus vitality. First case series in Argentina.

OBJECTIVE: To develop a sheep autologous uterus transplantation (UT) program with an innovative surgical technique and assess long term uterus vitality and animal survival. METHODS: A novel surgical technique consisting of the procurement of the complete uterus and the two ovaries, back table vascular reconstruction, and subsequent implantation in the same animal, performing only two arterial and two venous anastomoses. RESULTS: Four autologous transplantations were performed; anesthesia and surgery were well tolerated by all the animals without complications. Direct observation and Doppler US performed a week after UT and laparoscopy performed three months later confirmed uterus vitality. All animals were alive more than a year after transplantation. CONCLUSIONS: Our study was the first to describe a novel surgical technique for sheep uterus autologous transplantation in Latin America, showing long-term survival and uterus vitality.

Supramolecular solvent-based high-throughput sample treatment for monitoring phytohormones in plant tissues.

Quantification of endogenous hormones in plants is essential to understand their growth, development and response to biotic and abiotic stresses. However, it is challenging to develop high-throughput sample treatments from complex plant tissues containing low amounts of structurally unrelated and labile phytohormones while delivering clean and analyte-enriched extracts. In this paper we propose the use of supramolecular solvents (SUPRASs) made up or inverted hexagonal nanostructures of alkanols to address this challenge. The strategy was applied, as a proof of concept, to the quantification of stress-related phytohormones belonging to different categories (abscisic acid, salicylic acid, jasmonic acid, methyl jasmonate and 3-indoleacetic acid) in melon and pepper leaves. Sample treatment consisted in a single extraction-cleanup step involving the use of a low volume of SUPRAS (244 µL), the stirring (5 min) and centrifugation (15 min) of the sample at room temperature, and the direct analysis of the extract by liquid chromatography tandem mass spectrometry (LC-MS/MS). This high-throughput sample treatment method delivered excellent results for the target phytohormones regarding absolute recoveries (80-92%), method quantification limits (0.05-2 ng g-1), reproducibility (1-7%) and matrix effects (+13 to -31%), in both melon and pepper leaves, compared to reported methods based on repetitive solvent extraction, purification and solvent evaporation steps. The method was successfully applied to determine target hormones in melon and pepper plants for the evaluation of the effect of thermal stress. It was found that their concentration increased in the ranges 1.2-1.9 and 1.3-3.8 times in melon and pepper leaves, respectively, compared with control samples.

Supramolecular solvent-based high-throughput sample treatment platform for the biomonitoring of PAH metabolites in urine by liquid chromatography-tandem mass spectrometry.

Monohydroxylated polycyclic aromatic hydrocarbons (OH-PAHs), present in human urine at trace concentrations (viz. from ng L-1 to µg L-1), are considered the main biomarkers of human exposure to PAHs. In this work, we report a simple and high-throughput sample treatment platform to facilitate the biomonitoring of OH-PAHs by making it easier, greener and most cost-effective. This platform is based on the integration of analyte extraction and sample cleanup in a single step by the use of supramolecular solvents with restricted access properties (SUPRAS-RAM). The SUPRAS was spontaneously formed in situ in the urine by the addition of a colloidal suspension of decanoic acid in THF. Metabolites from naphthalene, fluorene, phenanthrene and pyrene were quantitatively extracted (absolute recoveries in the range 91-109%). Polysaccharides and proteins in the urine were excluded from extraction by physical and chemical mechanisms, which allowed the direct analysis of the SUPRAS extract by liquid chromatography tandem mass spectrometry. Absolute matrix effects for OH-PAHs were in the range 92-103%. Method quantification limits for OH-PAHs, without the need for evaporation of the SUPRAS extracts, were in the interval 1.0-6.7 ng L-1. The precision, evaluated in terms of repeatability and reproducibility, varied between 1.1 and 13.8%. The method was successfully applied to the analysis of urine from 16 smoking and non-smoking volunteers. Both analytical and operational features of this method make it suitable to evaluate human exposure to PAHs.

Bisphenol A and reproductive hormones and cortisol in peripubertal boys: The INMA-Granada cohort.

INTRODUCTION: Bisphenol A (BPA) is a well-known endocrine disrupting compound. Although several studies have investigated the effect of BPA exposure and reproductive hormones in humans, results have been inconsistent. OBJECTIVE: To explore the cross-sectional relationship between bisphenol A (BPA) exposure and reproductive hormones/cortisol among peripubertal boys. MATERIAL AND METHODS: Urinary BPA and serum hormones were assessed in 172 boys belonging to the INMA "Environment and Childhood" Granada birth cohort in their follow-up at 9-11years of age. BPA concentrations were quantified by liquid chromatography-mass spectrometry, and levels of serum total testosterone (TT), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and cortisol were measured by electrochemiluminescence immunoassay. RESULT(S): After adjustment for confounders, linear regression models showed that each natural-log unit increase in urinary BPA concentrations was associated with a 19% increase in geometric mean (GM) serum TT levels, and a 16% decrease in GM serum cortisol levels. When urinary BPA concentrations were categorized in tertiles, boys in the 3rd tertile showed 49% higher TT levels and 23% lower cortisol concentrations compared to boys in the 1st tertile. Additionally, urinary BPA concentrations were also significantly associated with higher TT:LH and TT:cortisol ratios, but not with serum LH or FSH levels. CONCLUSION(S): Our results suggest the possible endocrine disrupting potential of BPA during this important period of development. Although action at the testis or pituitary cannot be ruled out, our findings are compatible with a possible involvement of BPA at the adrenal gland, resulting in a differential production of androgens/cortisol. However, given the cross-sectional design of our study, the heterogeneous results reported in the literature, and the scant experimental research on BPA effects at the adrenal gland, the present findings should be interpreted with caution.