Complex chromosomal rearrangement—a lesson learned from PGS

Purpose

The aim of the study is to report a case of non-diagnosed complex chromosomal rearrangement (CCR) identified by preimplantation genetic screening (PGS) followed by preimplantation genetic diagnosis (PGD) which resulted in a pregnancy and delivery of healthy offspring.

Methods

A 29-year-old woman and her spouse, both diagnosed previously with normal karyotypes, approached our IVF-PGD center following eight early spontaneous miscarriages. PGS using chromosomal microarray analysis (CMA) was performed on biopsied trophectoderm. Fluorescence in situ hybridization (FISH), as well as re-karyotype, were performed on metaphase derived from peripheral blood of the couple. Subsequently, in the following PGD cycle, a total of seven blastocysts underwent CMA.

Results

A gain or loss at three chromosomes (3, 7, 9) was identified in six out of seven embryos in the first PGS-CMA cycle. FISH analysis of parental peripheral blood samples demonstrated that the male is a carrier of a CCR involving those chromosomes; this was in spite of a former diagnosis of normal karyotypes for both parents. Re-karyotype verified the complex translocation of 46,XY,t (3;7;9)(q23;q22;q22). Subsequently, in the following cycle, a total of seven blastocysts underwent PGD-CMA for the identified complex translocation. Two embryos were diagnosed with balanced chromosomal constitution. A single balanced embryo was transferred and pregnancy was achieved, resulting in the birth of a healthy female baby.

Conclusions

PGS employing CMA is an efficient method to detect unrevealed chromosomal abnormalities, including complicated cases of CCR. The combined application of array CGH and FISH technologies enables the identification of an increased number of CCR carriers for which PGD is particularly beneficial.

     

Splenectomy During Secondary Cytoreduction for Ovarian Cancer Disease Recurrence: Surgical and Survival Data

Background Ovarian cancer disease recurs predominantly in the abdomen, with the spleen usually involved as part of a vast spread of upper-abdominal disease or, less frequently, as an isolated site of disease recurrence. Very few reports are available in the literature on the outcome of patients subjected to splenectomy during secondary cytoreduction. The aim of this study was to identify prognostic factors and to review surgical and clinical data in order to identify those patients who would benefit the most from splenectomy during secondary cytoreduction.Methods This was a retrospective review of platinum-sensitive recurrent epithelial ovarian cancer patients who underwent splenectomy as part of secondary cytoreduction. Surgical and survival data were recorded.Results Twenty-four patients were identified. Multiple site disease recurrence was observed in 15 patients. The spleen was involved at the hilus in 12 patients; surface and intraparenchymal metastases were equally present. Optimal cytoreduction was achieved in all patients. At a median follow-up of 30 months, median progression-free and overall survival from the time of secondary surgery were 34 and 56 months, respectively. Overall survival was significantly correlated to residual disease at secondary surgery, disease-free survival, consolidation chemotherapy, and type of adjuvant therapy.Conclusions Splenectomy as part of secondary cytoreduction is a feasible and safe procedure. Secondary cytoreduction in selected groups of patients is confirmed to be associated with high long-term survival rates even when aggressive surgery of the upper abdomen is required.     

Supportive short-term family therapy by nursing staff in the inpatient unit: preventing dependence and rehospitalization in the acutely ill and suicidal adolescent

Adolescents with acute mental illness or suicidal behavior are almost always hospitalized for safety and evaluation purposes. The tendency towards long-term or repeated hospitalizations has many adverse effects such as dependency on the mental health care system and increased chronicity of illness. The causes for these phenomena may be prevented in the early stages of hospitalization. We suggest a therapeutic model of supportive short-term family therapy. The family therapy component aims to enhance the quality of interaction and the level of support among family members. The therapy component dealing with the individual targets the patient's anxiety symptoms and coping strategies, and focuses on return to a healthy state. The child is encouraged to return home to a supportive family as soon as the treatment team feels this to be advisable. This paper discusses a case which highlights how a patient reacts in crisis, and ways in which a supportive environment can help bring about therapeutic success with reduced hospitalization.     

Mitogen-activated protein kinase mediates luteinizing hormone-induced breakdown of communication and oocyte maturation in rat ovarian follicles

Resumption of meiosis, induced by LH, is preceded by the breakdown of gap junctional communication, which terminates the supply of cAMP from the somatic cells of the ovarian follicle to the oocyte. It has recently been shown that LH-induced reinitiation of meiosis is mediated by MAPK; however, the underlying molecular mechanism involved in the action of this enzyme remains unknown. We hypothesized that activation of MAPK interrupts junctional communication within the ovarian follicle, leading, in turn, to oocyte maturation. To test this hypothesis, we blocked the activation of MAPK by UO126, which specifically inhibits the MAPK signaling pathway. We analyzed junctional communication using three complementary methods: 1) patch-clamp analysis, which determined changes in the electrical coupling between two adjacent granulosa cells; 2) the scrape-loading technique, which monitored the spread of dyes through a granulosa cell layer; and 3) a metabolic coupling assay, which evaluated the transfer of radiolabeled uridine from the cumulus cells to the oocyte. We show, herein, that the somatic follicle cells, rather than the oocyte, activate MAPK immediately after their exposure to LH. Moreover, inhibition of LH-induced MAPK activation not only prevents oocyte maturation but also blocks the reduction in junctional communication. In addition, the appearance of the two phosphorylated forms of the gap junction protein, connexin 43, in response to LH, was avoided by UO126. We concluded that MAPK mediates LH-induced oocyte maturation by interrupting cell-to-cell communication within the ovarian follicle, possibly through phosphorylation of connexin 43.     

Metallothionein protein and minichromosome maintenance protein-2 expression in adrenocortical tumors

AimSome resected adrenal-confined adrenocortical carcinomas metastasize and others not. The present study was designed to evaluate the expression of metallothionein protein (MT) and minichromosome maintenance protein-2 (MCM2) in adrenocortical carcinomas and adrenocortical adenomas, and to test the correlation between this and adrenocortical carcinoma aggressiveness.Materials and methodsThe study comprised 14 patients operated on for adrenocortical carcinoma, 15 operated on for adrenocortical adenoma and 2 with normal adrenals. Hematoxylin-eosin staining was used for histological evaluation under light microscopy, and sequential sections were used for MCM2 and MT staining.ResultsIn normal adrenals, positive staining was weak for MT and zero for MCM2. Rates of positive staining for MT and MCM2 were significantly higher in adrenocortical carcinomas than in adrenocortical adenomas (P = 0.008 and P < 0.001, respectively). In adrenocortical carcinomas, a significant positive correlation was found between MCM2 staining and Weiss revisited score (P = 0.022) but not for Weiss score, and a significant positive correlation was found between MCM2 and mitotic rate on histology (P = 0.033). MCM2 but not MT staining was also shown to correlate significantly with stage IV carcinoma (P = 0.008 and P = 0.165, respectively).ConclusionMCM2 and MT are overexpressed in adrenocortical carcinoma, and MCM2 expression correlates significantly with metastatic disease.     

Reconstruction of Arabidopsis metabolic network models accounting for subcellular compartmentalization and tissue-specificity

Plant metabolic engineering is commonly used in the production of functional foods and quality trait improvement. However, to date, computational model-based approaches have only been scarcely used in this important endeavor, in marked contrast to their prominent success in microbial metabolic engineering. In this study we present a computational pipeline for the reconstruction of fully compartmentalized tissue-specific models of Arabidopsis thaliana on a genome scale. This reconstruction involves automatic extraction of known biochemical reactions in Arabidopsis for both primary and secondary metabolism, automatic gap-filling, and the implementation of methods for determining subcellular localization and tissue assignment of enzymes. The reconstructed tissue models are amenable for constraint-based modeling analysis, and significantly extend upon previous model reconstructions. A set of computational validations (i.e., cross-validation tests, simulations of known metabolic functionalities) and experimental validations (comparison with experimental metabolomics datasets under various compartments and tissues) strongly testify to the predictive ability of the models. The utility of the derived models was demonstrated in the prediction of measured fluxes in metabolically engineered seed strains and the design of genetic manipulations that are expected to increase vitamin E content, a significant nutrient for human health. Overall, the reconstructed tissue models are expected to lay down the foundations for computational-based rational design of plant metabolic engineering. The reconstructed compartmentalized Arabidopsis tissue models are MIRIAM-compliant and are available upon request.