Consequences of quality of reduction on osteosynthesis complications and quality of life in elderly patients with subtrochanteric fracture. / Consecuencias de la calidad de reducción sobre complicaciones en la osteosíntesis y calidad de vida en pacientes adultos de edad avanzada con fractura subtrocantérea.

OBJECTIVE: To compare results in terms of orthopaedic complications and quality of life in elderly patients with subtrochanteric fracture treated with intramedullary nailing according to fracture reduction status. PATIENTS AND METHODS: A prospective cohort study including 90 elderly patients with subtrochanteric fractures of the femur treated with a cephalomedullary nail, with a minimum 1-year follow up. The inclusion criteria were: aged 60 years or older, without severe cognitive dysfunction and independent ambulatory capability before the fracture. We defined 3different groups in relation to fracture reduction status: good, acceptable and poor, according to modified criteria from Baumgartner et al. We compared clinical and surgical characteristics and healthy quality of life, social function and mobility according to the EQ-5D, Jensen Index and Mobility Score of Parker and Palmer questionnaires. RESULTS: We found differences in time to union, better in the good reduction group (P=.002); need for open reduction, more frequent in the good reduction group (P<.001), and in postoperative complications, more frequent in the poor reduction group (P=.001). We found no significant differences between the 3groups regarding scores in quality of life, social function and mobility. CONCLUSIONS: Reduction in subtrochanteric fractures in older people is key to obtaining better clinic and surgical results, improving time to union and decreasing surgical complications. Exposure of the focus fracture seems to be a safe manoeuvre. Quality of life had substantially deteriorated n these patients, but a there was a tendency, although not statistically significant, for it to improve in patients after good surgical reduction.

Apoptosis mediated by phosphatidylserine externalization in the elimination of aneuploid germ cells during human spermatogenesis.

It has been described that aneuploidies trigger cell cycle checkpoints leading to apoptosis. The aim of this study was to assess the relationship between the presence of chromosomal abnormalities and apoptosis in germ cells and in Sertoli cells. Fourteen diagnostic testicular biopsies from infertile patients were processed following a sequential methodology, which included enzymatic disaggregation, apoptotic staining, cell sorting, cell fixation, and fluorescent in situ hybridization analysis. The chromosome constitution of germ cells (interphase pre-meiotic germ cells, meiotic figures, post-reductional germ cells, and spermatozoa) and Sertoli cells was evaluated in non-sorted and flow-sorted cell populations (apoptotic and viable). The mean percentage of aneuploidy was compared between the three fractions in each cell type using a Kruskal-Wallis test. If significant results were obtained, a two-by-two Chi-squared test was performed. There were significant differences between the apoptotic fraction and the viable and non-sorted fractions in the pre-meiotic germ cells (p < 0.01). In the remaining cell types, no association between the presence of aneuploidy and apoptotic processes was observed, even in the case of post-reductional germ cells in which we detected the highest rates of aneuploidy regardless of the fraction analyzed. From our data, it can be inferred that most of the aneuploid post-reductional germ cells are efficiently removed from the testicular epithelium without differentiating into spermatozoa. Our results suggest that the elimination of aneuploid testicular epithelial cells is triggered by different mechanisms. Accordingly, the cellular elimination of aneuploid germ cells beyond the blood-testis barrier does not involve phosphatidylserine externalization.

Hidden mosaicism in patients with Klinefelter's syndrome: implications for genetic reproductive counselling.

BACKGROUND: Most individuals with Klinefelter's syndrome (KS) are azoospermic but residual foci of spermatogenesis have been observed in some patients. However, no consistent predictive factors for testicular sperm extraction success have been established and mosaicism could be a factor to investigate. In this study, we have assessed the degree of mosaicism in somatic and germinal tissues in KS, the meiotic competence of 47,XXY germ cells and the aneuploidy rate of post-reductional cells. METHODS: Five patients with KS previously diagnosed as pure 47,XXY have been studied. Samples from four donors were processed as controls. The chromosome constitution of lymphocytes, buccal mucosa and testicular tissue was assessed by interphase fluorescence in situ hybridization for chromosomes X, Y and 18. In meiotic figures, sex chromosome number and pairing was confirmed. RESULTS: 46,XY cell lines were observed in all patients and tissues analysed. The degree of mosaicism (mean ± SD) differed among tissues (lowest in lymphocytes: 4.8 ± 2.5%; highest in Sertoli cells: 42.3 ± 11.1%). Meiotic figures were found in three cases (KS1, KS2 and KS5), all of them showed an XY complement. Hyperhaploid post-reductional cells were found in all patients (range: 3.3-36.4%) and increased rates versus controls (P< 0.05) were observed. CONCLUSIONS: Diagnosis of homogeneous KS based on lymphocyte karyotyping should be contrasted in other tissues. Mucosa cells could help to better approximate the degree of germ cell mosaicism. Our results indicate that 47,XXY germ cells are not meiotically competent. Increased post-reductional aneuploidy rate is related to meiotic errors in 46,XY cells. Appropriate genetic counselling is recommended in KS.

Complete meiosis from human induced pluripotent stem cells.

Gamete failure-derived infertility affects millions of people worldwide; for many patients, gamete donation by unrelated donors is the only available treatment. Embryonic stem cells (ESCs) can differentiate in vitro into germ-like cells, but they are genetically unrelated to the patient. Using an in vitro protocol that aims at recapitulating development, we have achieved, for the first time, complete differentiation of human induced pluripotent stem cells (hiPSCs) to postmeiotic cells. Unlike previous reports using human ESCs, postmeiotic cells arose without the over-expression of germline related transcription factors. Moreover, we consistently obtained haploid cells from hiPSCs of different origin (keratinocytes and cord blood), produced with a different number of transcription factors, and of both genetic sexes, suggesting the independence of our approach from the epigenetic memory of the reprogrammed somatic cells. Our work brings us closer to the production of personalized human gametes in vitro.