R337H mutation of the TP53 gene as a clinical marker in cancer patients: a systematic review of literature.

The germline R337H mutation of the TP53 gene has been associated with the development of many tumor types. This systematic review of literature investigated the association between the R337H mutation and the patients' family history and its predictive and prognostic value in cancer. Data were collected from articles archived in the PubMed, LILACS, MEDLINE, IBECS, and SciELO databases. The systematic review of literature was performed on 12 selected articles, describing a total of 175,462 individuals tested for the R337H mutation, including 1548 individuals with cancer and 118 individuals with a family history of Li-Fraumeni and Li-Fraumeni-like syndrome. Eight studies showed an association between the mutation and a family history of cancer in 411 patients, including 390 cases of cancer among family members. Patients with the homozygous mutant genotype experienced cancer recurrence, progressive disease, secondary cancer, and a short survival rate. Heterozygous patients showed a better response to treatment and increased survival rates than did patients with the homozygous mutant genotype from newborns to adult patients. In conclusion, the R337H mutation has significant predictive and prognostic value and is associated with tumorigenesis of the adrenal cortex.

Analysis of the BRAF V600E mutation in primary cutaneous melanoma.

BRAF V600E is the most common mutation in cutaneous melanomas, and has been described in 30-72% of such cases. This mutation results in the substitution of valine for glutamic acid at position 600 of the BRAF protein, which consequently becomes constitutively activated. The present study investigated the BRAF V600E mutation frequency and its clinical implications in a group of 77 primary cutaneous melanoma patients treated in a cancer reference center in Brazil. Mutation analysis was accomplished by polymerase chain reaction, restriction fragment length polymorphism, and automated DNA sequencing. The chi-squared and Fischer exact tests were used for comparative analyses. The BRAF V600E mutation was detected in 54/77 (70.1%) melanoma subjects. However, no statistically significant association was found between the presence of the mutation and clinical or prognostic parameters. Our results demonstrated that the BRAF V600E mutation is a common event in melanomas, representing an important molecular target for novel therapeutic approaches in such tumors.

Effect of prostaglandin E2, lipopolysaccharide, IFN-gamma and cytokines on the generation and function of fast-DC.

BACKGROUND: Recent reports have described a new strategy for differentiation and maturation of monocyte-derived DC within only 48 h of in vitro culture (fast-DC). We compared the ability of various maturation stimuli with the generation of Ag-specific T-cell responses and generation of functional fast-DC. METHODS: CD14+ cells were treated with GM-CSF and IL-4 for 1 day to generate immature DC, and were then matured with either inflammatory cytokines or a combination of lipopolysaccharide (LPS) and INF-gamma. Mature DC were then used to study the effect of prostaglandin E2 (PGE2) on the stimulatory function of fast-DC. RESULTS: fast-DC were CD14- and expressed mature DC surface markers, and maintained this phenotype after withdrawing the cytokine from culture. Treatment of fast-DC with a combination of LPS and INF-gamma promoted the maturation of highly uniform fast-DC. The T-cell proliferative response to DC was enhanced by inclusion of PGE2 in the MCM-mimic (TNF-a, IL-1 a, IL-6, PGE2) cocktail. DISCUSSION: fast-DC are very effective; they not only reduce the labor, cost and time required for in vitro DC development, but may also represent a model more closely resembling DC differentiation from monocytes in vivo.

Estimation of the tissue composition of the tumour mass in neuroblastoma using segmented CT images.

Neuroblastoma is the most common extra-cranial, solid, malignant tumour in children. Advances in radiology have made possible the detection and staging of the disease. Nevertheless, there is no method available at present that can go beyond detection and qualitative analysis, towards quantitative assessment of the tissue composition of the primary tumour mass in neuroblastoma. Such quantitative analysis could provide important information and serve as a decision-support tool to the radiologist and the oncologist, result in better treatment and follow-up and even lead to the avoidance of delayed surgery. The problem investigated was the improvement of the analysis of the primary tumour mass, in patients with neuroblastoma, using X-ray computed tomography (CT) images. A methodology was proposed for the estimation of the tissue content of the mass: it comprised a Gaussian mixture model for estimation, from segmented CT images, of the tissue composition of the primary tumour. To demonstrate the potential of the method, the results are presented of its application to ten CT examinations of four patients. The method provides quantitative information, and it was observed that the tumour in one of the patients reduced from 523 cm3 to 81 cm3 in volume, with an increase in calcification from about 20% to about 88% of the tumour volume, in response to chemotherapy over a period of five months. Results indicate that the proposed technique may be of considerable value in assessing the response to therapy of patients with neuroblastoma.

Generation of Ag-specific cytotoxic T lymphocytes by DC transfected with in vitro transcribed influenza virus matrix protein (M1) mRNA.

BACKGROUND: Application of DC transfected with tumor Ag RNA is promising for DC-based tumor immunotherapy. In this study, Ag-specific cytotoxic T lymphocytes (CTL) were generated by priming lymphocytes with DC transfected with in vitro transcribed (IVT) influenza virus matrix protein M1 (M1) mRNA. METHODS: Human UC blood-CD34+ cell-derived DC were transfected with IVT mRNA encoding either the enhanced green fluorescence protein (EGFP), or M1 by square-wave electroporation. DC were confirmed to have typical morphology and phenotype. DC transfected with IVT EGFP mRNA were analyzed with the FACScan flow cytometer, to confirm the efficiency of this transfection method. On Days 7, 14, 21 and 28 after the start of DC culture, DC were harvested and electroporated with M1 mRNA. The transfected DC were co-cultured with autologous UC blood CD34- cells. One week after the fourth priming of autologous CD34 negative cells with M1 mRNA electroporated DC, Ag-specific CTL activity was evaluated. To prepare target cells, M1 mRNA was added to autologous DC 48 h prior to CTL assays. RESULTS: Our CTL assays results indicate that UC blood CD34+ cell-derived DC transfected with M1 mRNA by electroporation stimulated Ag-specific CTL responses that are capable of recognizing and lysing autologous DC loaded with M1 mRNA. M1 mRNA transfected DC-primed CTL showed a significant cytotoxic activity against M1 mRNA loaded autologous DC, while nearly baseline cytotoxic activity was recorded for the M1 mRNA unloaded DC. DISCUSSION: Our results showed that mRNA-transfected DC are potent stimulators of T-cell immunity in vitro. In addition, mRNA-loaded DC can function as targets in CTL cytotoxicity assays, which offer a practical substitute for tumor cells in assays to test the immunological effects of specific Ags.

Dipole defects in Al2O3:Mg,Cr.

In this work, dipole defects are investigated applying the thermally stimulated depolarisation currents (TSDC) technique. The TSDC spectra of Al2O3 doped with Mg and Cr show two bands centred at 230 K and 250 K, respectively. The maximum intensity of the bands increases linearly with the polarisation field, a typical behaviour of defects with dipole origin. An increase of the band at 250 K with gamma irradiation has been observed and a thermal decrease of the bands for heat treatments between 1000 K and 1400 K. Above this temperature the bands are partially recovered. Impurity neutron activation analysis shows that magnesium. chromium and iron content varies from 15 to 60 ppm. Optical absorption (AO) measurements show a broad band centred in 2.6 eV (21000 cm(-1)) associated with trapped holes localised on an O- ion adjacent to a cation site which is deficient in positive charge. It has been assumed that a substitutional Mg2+ ion occupies the cation site near a trapped hole on one of the six oxygen ions surrounding the magnesium impurity giving rise to the dipole responsible for the observed TSDC bands. Calculations carried out through defect simulation methods confirm that the probability of Al3+ being replaced by Mg2+ is higher than Mn2+, Co2+, Fe2+ and Cr2+.

Generation of dendritic cells from leukaemia cells of a patient with acute promyelocytic leukaemia by culture with GM-CSF, IL-4 and TNF-alpha.

Dendritic cells are potent antigen-presenting cells derived from CD34+ haemopoietic stem cells. Dendritic cells have been reported to be generated from cells in granulocytic lineage as well as monocytes, blood dendritic cell precursors and lymphoid progenitors. In order to explore the differentiation pathway of dendritic cells from granulocytic cells and the applicability of leukaemia-derived dendritic cells for anti-leukaemic immunotherapy in acute leukaemia of granulocytic origin, we tried to generate dendritic cells from leukaemia cells of a patient with acute promyelocytic leukaemia (APL). Leukaemia cells were cultured with GM-CSF, IL-4 and TNF-alpha for 10 days. Azurophilic granule-containing cells with marked cytoplasmic projections were generated in the culture. FACS analysis of these cultured cells revealed the generation of CD1a+, CD83+, CD80+, CD86+, CD40+ and HLA-DR+ cells. The leukaemic origin of these dendritic-like cells was demonstrated by in situ hybridization of magnetic-bead-sorted CD1a+ dendritic cells using the DNA probes of t(15;17). Cells generated by culturing leukaemia cells were demonstrated to have a potent antigen-presenting function in allogeneic mixed leucocyte cultures. These findings show the plausibility of the previously reported pathway of dendritic cell maturation through granulocytic cells and suggest the possibility of anti-leukaemic immunotherapy using leukaemia-derived dendritic cells even in patients with acute promyelocytic leukaemia.

Observations on the commercial production of ostrich (Struthio camelus) in the United Kingdom: incubation.

Two sets of ostrich eggs (60 and 120 eggs) were imported into the United Kingdom under class 1 quarantine restrictions. Single stage incubation was carried out and the eggs were weighed before and during incubation in order to control weight loss. In the two hatches the weight losses during the incubation of viable eggs were 13.4 per cent and 11.4 per cent, respectively. The development of the eggs was followed by candling and although only dark shadows were observed a pattern could be recognised. For the first set of eggs the average length of incubation was 45.9 days with an interval of 12.7 hours between pipping and hatching. The second set of eggs was incubated at higher temperatures than the first and the main incubation period was 43.3 days with hatching 19.2 hours after pipping. Larger eggs took longer to hatch.

Observations on the commercial production of ostrich (Struthio camelus) in the United Kingdom: rearing of chicks.

Two sets of ostrich eggs (60 and 120 eggs) were imported into the United Kingdom under class 1 quarantine restrictions. The eggs were incubated and observations were made on the growth, survival and sex ratio of the chicks hatched. The chicks decreased in weight for five days after hatching before they began a sustained period of exponential growth. They reached a liveweight of 4 kg five weeks after hatching. Female chicks grew significantly faster than male chicks. The survival rates of the chicks to three months of age were 66.7 per cent and 78.3 per cent, respectively, for the two sets of eggs, and mortality restricted mainly to the first four weeks of rearing. All the birds which died showed poor rates of growth before they died. The sex ratio of both groups was skewed 2:1 towards males.