Preimplantation genetic diagnosis of chromosome abnormalities: implications from the outcome for couples with chromosomal rearrangements

OBJECTIVES: Chromosomal rearrangements can lead to infertility or repeated spontaneous or induced abortions. The use of preimplantation genetic diagnosis (PGD) allows the selected transfer of chromosomally balanced embryos. The aim of this study was to carry out detailed analysis of the outcome of 11 PGD cycles for 8 patients carrying various chromosomal rearrangements. METHODS: Patients underwent routine in vitro fertilisation with biopsy of embryos on day 3. Specific fluorescent in situ hybridisation protocols were developed for each couple. Embryo transfer was possible in all 11 cycles. RESULTS: The outcome was four pregnancies, leading to three live births and one biochemical pregnancy. Post-zygotic mosaicism was detected in 75% of untransferred embryos, the majority of which were chaotic. Detailed follow-up and analysis provided evidence for the co-existence of chromosomally balanced and abnormal cells in six embryos. The mechanisms involved included chromosome breakage and loss of material. CONCLUSIONS: Biopsy and analysis of two blastomeres, where possible, reduced the risk of misdiagnosis in cases of balanced/aneuploid mosaics. The three live births achieved for the eight couples treated in this series, despite the poor history in almost all cases, is further proof that a policy of biopsying two cells from embryos consisting of six or more cells and a single cell from four- or five-cell embryos is compatible with a positive outcome.     

Comparison between rules-based human immunodeficiency virus type 1 genotype interpretations and real or virtual phenotype: concordance analysis and correlation with clinical outcome in heavily treated patients

We compared 2 rules-based genotype interpretation systems and real or virtual phenotype through a retrospective analysis of a prospective trial. Genotypes were determined with VircoGEN II (VIRCO) and were interpreted with either RetroGram 1.4 or TRUGENE HIV-1 (guidelines 3.0) or original virtual phenotype (Virtual Phenotype; VIRCO), as available in the year 2000. Among 188 human immunodeficiency virus (HIV) type 1 isolates, overall concordance (kappa agreement) was observed for the 2 rules-based systems, whereas striking discordances were noted between them and real and virtual phenotype interpretations for stavudine, didanosine, zalcitabine, abacavir, and amprenavir (kappa<0.4). Clinical evaluation of a subset of 173 patients showed that both rules-based sensitivity scores were independently associated with HIV RNA loads <400 copies/mL at week 16 of during-treatment analysis (TRUGENE: odds ratio [OR], 2.90; 95% confidence interval [CI], 1.52-5.52; P=.001; RetroGram: OR, 2.34; 95% CI, 1.21-4.55; P=.012), whereas, in contrast to real or virtual phenotype, interpretations according to biological cut-offs were not (OR, 1.91; 95% CI, 0.77-4.76; P=.162).     

Iron supplementation in a child with severe burns

Non-intentional burns are still a frequent cause of morbidity in children despite the implementation of preventive campaigns. Children with severe burns (more than 10 % of the body surface) may require iron supplementation, especially in the post-stress or recovery phase. We report the case of a 15-month-old boy, who suffered polytrauma and burns to 20 % of the body surface. A dermal graft was performed eight days after his admission to the pediatric burns unit, which required transfusion of packed red blood cells. On day 15 hemoglobin value was 9.3 g/dL, the reticulocyte hemoglobin content was reduced to 26.2 pg (normal value: 27.2-30.3 pg) and the reticulocyte mean corpuscular value was increased to 124 fl (normal value: 101-108 fl). These findings were interpreted as a functional iron insufficiency, despite the increased ferritin value (219 ng/mL). We highlight the satisfactory outcome of the different hematologic values, with the development of reticulocyte crisis and normalization of hematological parameters after administration of 0.5 mg/kg/day of ferrous sulfate, plus iron corresponding to the follow-up milk formula and a regular diet for the child's age.     

Physiological effects of translaryngeal open ventilation in patients with restrictive respiratory disorders

OBJECTIVE: To compare the efficacy of pressure-controlled ventilation (PCV) delivered through a conventional endotracheal tube with the same ventilatory mode using a small-size tube with the cuff left deflated (translaryngeal open ventilation: TLOV). SETTING: A medical-surgical intensive care unit (ICU). DESIGN: Prospective physiological study. PATIENTS: Thirteen consecutive patients with restrictive neuromuscular and thoracic respiratory disorders ventilated in pressure-controlled mode. INTERVENTIONS: The standard tube was removed and a microlaryngeal tube (i.d. 4 mm, o.d. 6 mm, length 380 mm) was inserted with the cuff left deflated. PCV was increased to match the tracheal pressure measured during conventional ventilation. Arterial blood gases were measured before, 1 h and 20 h after initiating TLOV. A patient comfort score was measured by a visual analogue scale during conventional ventilation and 20 h after initiating TLOV (0= very bad, 1= bad, 2= quite bad, 3= sufficient, 4= good, and 5= very good). RESULTS: Inspiratory pressure was significantly increased from 16+/-5 cmH(2)O to 68+/-13 cmH(2)O after 1 h and to 65+/-12 cmH(2)O after 20 h to match the tracheal pressure measured during conventional ventilation (CV) (p<0.005). No statistically significant differences were found in arterial blood gases and patient's respiratory rate before and after 1 and 20 h of TLOV. The comfort score was 1.3+/-0.4 and 3.6+/-0.4 during CV and TLOV, respectively, on a scale from 0 to 5 (p<0.002). CONCLUSION: This study indicates that, in selected patients, TLOV was as efficient as conventional PCV.     

RETMEN2A and RETMEN2B oncoproteins are targets of PP1 inhibitor

Medullary thyroid carcinoma (MTC) responds very poorly to chemotherapy. Mutations in the RET gene are critical for MTC pathogenesis. RET therefore represents a rational target for the development of novel MTC therapies. The accumulation of evidence from laboratory studies strongly suggests that PP1 inhibitor is a cytostatic agent for cells expressing RET oncoproteins. PP1 functions as a potent and selective inhibitor of RET oncoprotein phosphorylation, promoting its proteasomal degradation.     

Disease stage variation in CD4+ and CD8+ T-cell reactivity to the receptor tyrosine kinase EphA2 in patients with renal cell carcinoma

We have evaluated CD8+ and CD4+ T-cell responses against a new tumor-associated antigen, the receptor tyrosine kinase EphA2, which is broadly expressed in diverse cancer histologies and is frequently overexpressed in advanced stage/metastatic disease. We report herein that EphA2 is overexpressed in renal cell carcinoma (RCC) cell lines and clinical specimens of RCC, and find that the highest levels of EphA2 are consistently found in the most advanced stages of the disease. We identified and synthesized five putative HLA class I-binding and three class II-binding peptides derived from EphA2 that might serve as targets for immune reactivity. Each peptide induced specific, tumor-reactive CD8+ or CD4+T-cell responses as measured using IFN-gamma enzyme-linked immunospot assays. The EphA2 peptides elicited relatively weak responses from CD8+ T cells derived from normal healthy volunteers or from RCC patients with active disease. In marked contrast, immune reactivity to EphA2-derived epitopes was greatly enhanced in CD8+ T cells that had been isolated from patients who were rendered disease-free, after surgery. Furthermore, enzyme-linked immunospot analyses demonstrated prominent EphA2-restricted T-helper 1-type CD4+ T cell activity in patients with early stage disease, whereas T-helper 2-type and T regulatory-type responses predominated in patients with more advanced forms of RCC. These data suggest that the immune system of cancer patients actively monitors EphA2-derived epitopes, and that the magnitude and character of T-cell responses to EphA2 epitopes may convey much-needed predictive information about disease stage and outcome.     

The dog mast cell tumour as a model to study the relationship between angiogenesis,mast cell density and tumour malignancy

Substantial experimental data suggest that tumour progression is associated with angiogenesis and that increase in microvessel density (MVD) is associated with increase in mast cells density (MCD). Dog mast cell tumour (MCT) is common in dog with an incidence much higher than that found in human and in both species several common biological and clinical characteristics have been demonstrated. To evaluate the role of angiogenesis in progression of this tumour and to correlate MVD and MCD, in this study a series of 78 MCT was investigated. Serial sections obtained from biopsy specimens were processed with toluidine blue staining, specific for MC identification, and by immunohistochemistry using a polyclonal antibody anti factor VIII-related antigen (FVIII-RA), used as an endothelial marker, and MVD and MCD were determined. Results showed that MVD was significantly higher in poorly differentiated (G3) MCTs than in intermediate (G2) and well differentiated (G1) MCTs and that MCD and MVD were significantly correlated in G3, but not in G1 and G2 subgroups. These data indicate that angiogenesis and MCD are significantly correlated in MCTs progression.     

Cognitive-existential group psychotherapy for women with primary breast cancer: a randomised controlled trial

BACKGROUND: We conducted a randomised, controlled trial of cognitive-existential group therapy (CEGT) for women with early stage breast cancer receiving adjuvant chemotherapy with the aim of improving mood and mental attitude to cancer. METHODS: Women were randomised to 20 sessions of weekly group therapy plus 3 relaxation classes or to a control arm receiving 3 relaxation classes. Assessments, independently done at baseline, 6 and 12 months, included a structured psychiatric interview and validated questionnaires covering mood, attitudes to cancer, family relationships, and satisfaction with therapy. RESULTS: Three hundred and three of 491 (62%) eligible patients participated over 3 years. Distress was high pre-intervention: 10% were diagnosed as suffering from major depression, 27% from minor depression and 9% from anxiety disorders. On an intention-to-treat analysis, there was a trend for those receiving group therapy (n=154) to have reduced anxiety (p=0.05, 2-sided) compared to controls (n=149). Women in group therapy also showed a trend towards improved family functioning compared to controls (p=0.07, 2-sided). The women in the groups reported greater satisfaction with their therapy (p<0.001, 2-sided), appreciating the support and citing better coping, self-growth and increased knowledge about cancer and its treatment. They valued the CEGT therapy. Overall effect size for the group intervention was small (d=0.25), with cancer recurrence having a deleterious effect in three of the 19 therapy groups. Psychologists as a discipline achieved a moderate mean effect size (d=0.52). CONCLUSION: CEGT is a useful adjuvant psychological therapy for women with early stage breast cancer. Interaction effects between group members and therapists are relevant to outcome. Group-as-a-whole effects are powerful, but the training and experience of the therapist is especially critical to an efficacious outcome.     

Angiogenesis and angiogenesis inhibitors: a new potential anticancer therapeutic strategy

Tumor cells cannot grow as a mass above 2 to 3 mm3 because diffusion is insufficient for oxygen and glucose requirements, unless the tumor induces a blood supply. This mechanism of induction of a new blood supply from pre-existing vascular bed is called angiogenesis. Furthermore, tumor invasiveness and metastasis require neovascularization. In fact, recent published studies suggest that acquisition of the angiogenic phenotype is a common pathway for tumor progression and neovascularization is linked with other molecular steps leading to tumor progression. Angiogenic process is a complex multi-step cascade under the control of positive and negative soluble factors. A paracrine interaction occurs between tumor and endothelial cells. Angiogenesis involves: endothelial cell proliferation, migration and tubule formation with associated changes in the extra-cellular matrix, allowing subsequent new vessel growth toward the tumor. Each of the above steps may represent a target for antiangiogenic therapy. Antiangiogenesis is to be distinguished from direct targeting and destruction of tumor vasculature (vascular targeting). Inhibition of angiogenesis represents one of the more promising, new approaches, to anticancer treatment and its already in early clinical trials. This review takes into consideration: (i) the biological mechanism underlining angiogenesis process; (ii) the method to assess tumor angiogenesis activity; (iii) inhibition of angiogenesis as an anticancer therapy; (iv) the methodology for the clinical development of angiogenesis inhibitors, that should be considered biological response modifiers; (v) some angiogenesis antagonists that are in development and leader compounds that are under clinical trial.