Cell proliferation in breast cancer is a major determinant of clinical outcome in node-positive but not in node-negative patients.

The growth rate of a tumor cell population depends on two major factors: the percentage of proliferating cells (cell growth fraction) and the rapidity of their duplication (cell proliferation rate). The authors evaluated the prognostic and predictive value of both kinetics parameters in a large series of breast cancer patients (n=504). The cell growth fraction was determined by MIB-1 immunostaining, the cell proliferation rate by AgNOR analysis. Ki-67 LI (labeling index) and AgNOR area were significantly associated with histotype, histologic grade, tumor size, estrogen/progesterone receptor status, patient age, and lymph node involvement (P<0.005). In the entire series of patients, both kinetics variables were significantly and independently associated with the clinical outcome, but their prognostic relevance was quite different when node-negative and node-positive patients were considered separately. Although in node-positive patients Ki-67 LI and AgNOR area were the unique independent predictors of disease-free and overall survival, they were excluded by the multivariate Cox model in node-negative patients, where only tumor size and estrogen receptor status retained a significant P-value. These results show that in breast carcinoma the cell growth fraction and the cell proliferation rate have a different prognostic impact with respect to the lymph node status and are major determinants of clinical outcome in node-positive patients only. Within this subgroup, the rapidity of cell proliferation as assessed by AgNOR analysis also served as a sensitive predictor of the response to adjuvant treatments.     

K-ras gene mutational analysis supports a monoclonal origin of biphasic pleomorphic carcinoma of the lung.

We investigated 27 pleomorphic carcinomas of the lung for exon 1 K-ras gene mutations using polymerase chain reaction-single-strand conformation polymophism analysis and direct sequencing. All pleomorphic carcinomas were biphasic, that is, composed of an adeno-, squamous- or large-cell-carcinomatous component associated with a spindle- and/or giant-cell component. Of 27 cases, six (22%) showed K-ras codon 12 mutations, which is a figure higher than that previously reported on in pure sarcoma-like pleomorphic carcinomas. Five tumors displayed the same mutation in both the epithelial and the sarcomatoid components, whereas in one tumor the mutation was restricted to the epithelial component. All mutations occurred in smokers, and were transversions, including GGT (glycine) to TGT (cysteine) change in two cases, to GCT (alanine) in two and to GTT (valine) in two. No significant relationships were found between the occurrence and type of mutations and patients' survival or any other clinicopathological variable, suggesting that K-ras mutations are early events in the development of these tumors. Our results indicate that most, though not all, biphasic pleomorphic carcinomas of the lung are monoclonal in origin, and that cigarette smoking may have a causative role in the development of K-ras alterations in these tumors, as all mutations are transversions.     

Involvement of CD80 in the generation of CD4+ cytotoxic T cells

CD4+ T cells can exert different effector functions, which are partly distinguishable by the secretion of different cytokines, namely by either IFN-γ, IL-2 and lymphotoxins for Th1-like or IL-4, IL-5, IL-10 and IL-13 for Th2-like T cells. Th1-like T cells can exert cytotoxic functions, too. The cytokinetic phenotype of an activated T cell clone (TCC) is mainly influenced by the cytokinetic pattern of the microenvironment where it was activated. However, the interaction between certain adhesion molecules (i.e. CD28-CD80 and CD28-CD86) may also have an influence on the functionality of the reactive T cell. On the contrary, the requirements for the induction of CD4+ cytotoxic T cells (CD4+CTLs) are not well understood. We have focused this review on studies investigating the development of CD4+ T cells with cytotoxic effector functions. In particular, we discuss here whether the type of antigen-presenting cells (APCs) and the distinct expression of important adhesion molecules like CD80 and CD86 may influence the generation of CD4+ CTLs. Among a large panel of APCs only dendritic cells and TCCs are able to induce cytotoxicity. The level of CD80, but not of CD86, present on the APCs appears to be crucial for the induction of CD4+ CTLs.     

Behavior of the bone-titanium interface after push-in testing: a morphological study

Fourteen titanium dental implants (Tioblast) were implanted singly in the proximal tibia of New Zealand rabbits for 120 days. A bone defect was surgically produced and filled with Bio-Oss around six of these implants. After the animals were sacrificed and their organs harvested, bone segments were fixed and methacrylate embedded after the push-in test had been performed. Microradiography was performed on longitudinal sections of the implants, whereas scanning electron microscope analysis was performed on the remaining embedded half-implants using secondary electrons only. The results showed that the implants were apically and coronally surrounded by bone, whether Bio-Oss was used or not. Fractures were evident through the newly formed bone and between the pre-existing and newly formed bone. Some fracture lines propagated through the bone and stopped at the implant surface without continuing along the bone-titanium interface. Detachment between the implant and the bone occurred at the coronal extremity of the implants and along its cervical region. These results highlight the fact that the bone-titanium interface has a high resistance to loading. It exhibited greater resistance than the newly formed bone and seems to behave in a manner similar to the cement lines of osteons.     

Transmural distribution of antioxidant defences and lipid peroxidation in the rabbit left ventricular myocardium

The left ventricular subendocardial and subepicardial layers of six perfused rabbit hearts were tested for enzymatic and non-enzymatic antioxidant defences and for lipid peroxidation. The subendocardium showed significantly lower catalase activity and contents of non-protein thiol compounds and vitamin E associated with a higher degree of lipid peroxidation. The activities of Cu,Zn- and Mn-superoxide dismutases, glutathione reductase, -glutamylcysteine synthetase and -glutamyl transpeptidase showed no significant transmural differences, and Se-independent glutathione peroxidase activity was not detectable in either layer. Comparable results were observed in another group of six unperfused rabbit hearts. In five H₂O₂-perfused rabbit hearts, lipid peroxidation was higher, and myocardial creatine phosphokinase activity lower, in the subendocarium than in the subepicardium. In this group, only the subendocardium had significantly higher lipid peroxidation levels than the control hearts. Thus, a lower antioxidant capacity and a greater oxidative stress are present in the rabbit subendocardium. These findings could provide insight into the problem of subendocardial vulnerability to free radical-mediated processes, such as occurs in ischaemia-reperfusion injury.     

Compartmental Analysis of Breathing in the Supine and Prone Positions by Optoelectronic Plethysmography

Optoelectronic plethysmography (OEP) has been shown to be a reliable method for the analysis of chest wall kinematics partitioned into pulmonary rib cage, abdominal rib cage, abdomen, and right and left side in the seated and erect positions. In this paper, we extended the applicability of this method to the supine and prone positions, typically adopted in critically ill patients. For this purpose we have first developed proper geometrical and mathematical models of the chest wall which are able to provide consistent and reliable estimations of total and compartmental volume variations in these positions suitable for clinical settings. Then we compared chest wall (CW) volume changes computed from OEP( V _CW) with lung volume changes measured with a water seal spirometer (SP) ( V _SP)in 10 normal subjects during quiet (QB) and deep (DB) breathing on rigid and soft supports. We found that on a rigid support the average differences between V _SP and V _CW were –4.2% ± 6.2%, –3.0% ± 6.1%, –1.7% ±7.0%, and –4.5% ± 9.8%, respectively, during supine/QB, supine/DB, prone/QB, and prone/DB. On the soft surface we obtained –0.1% ± 6.0%, –1.8% ± 7.8%, 18.0% ± 11.7%, and 10.2% ± 9.6%, respectively. On rigid support and QB, the abdominal compartment contributed most of the V _CW in the supine (63.1% ± 11.4%) and prone (53.5% ± 13.1%) positions. V _CW was equally distributed between right and left sides. In the prone position we found a different chest wall volume distribution between pulmonary and abdominal rib cage (22.1% ± 8.6% and 24.4% ± 6.8, respectively) compared with the supine position (23.3% ± 9.3% and 13.6% ± 3.0%). © 2001 Biomedical Engineering Society. PAC01: 8763Lk, 8719Uv     

Chromosomal localization of a highly repeatedEcoRI DNA fragment inMegoura viciae (Homoptera,Aphididae) by nick translation and fluorescencein situ hybridization

To investigate the genome of the aphidMegoura viciae at molecular level, we have studied total DNA by agarose gel electrophoresis after cleavage with different restriction endonucleases.EcoRI digestion produced a highly repeated DNA fragment, about 600 bp long. The contribution of thisEcoRI element to the total genome ofM. viciae was estimated at about 6% by means of densitometric scanning of agarose gel photographs. The chromosomal localization of this fragment, investigated by fluorescentin situ hybridization (FISH), constantly showed one large and two narrower fluorescent bands located on the X chromosome, all corresponding to C-positive heterochromatic areas. These results are in full accordance with the data obtained byin situ nick translation experiments carried out afterEcoRI digestion, and clearly demonstrate that a substantial amount ofM. viciae heterochromatin consists ofEcoRI fragments which are mainly located on the X chromosome. Using theEcoRI restriction fragment as a molecular probe may prove to be a practical tool for the investigation of taxonomic and evolutionary relationships in this group of insects.accepted for publication by J. S. (Pat) Heslop-Harrison     

Lack of association between angiotensin converting enzyme polymorphism and sporadic Alzheimer's disease

Epidemiological and pathogenetic evidences suggest a strong association between vascular risk factors and sporadic Alzheimer's disease (sAD). In agreement with the vascular hypothesis of AD, the role of various candidate genes for atherosclerosis has been investigated, leading to conflicting results. In order to clarify the significance of angiotensin-converting enzyme (ACE) gene insertion (I)/deletion (D) polymorphism in a group of patients with sAD, we conducted a case-control study including 149 cases and 149 age and sex matched controls. All subjects were genotyped for ACE and Apolipoprotein E (APOE). There were no significant differences in ACE genotype or allele frequencies between cases and controls, even after stratification for APOE4 carrier status. Our data suggest that the ACE I/D polymorphism is not associated to genetic susceptibility in sAD patients.     

CD34+ Cord Blood Cell-Transplanted Rag2−/− γc−/− Mice as a Model for Epstein-Barr Virus Infection

Recent studies suggest that Epstein-Barr virus (EBV) can infect naïve B cells, driving them to differentiate into resting memory B cells via the germinal center reaction. This hypothesis has been inferred from parallels with the biology of normal B cells but has never been proven experimentally. Rag2^−/− γ_c^−/− mice that were transplanted with human CD34⁺ cord blood cells as newborns were recently shown to develop human B, T, and dendritic cells, constituting lymphoid organs in situ. Here we used this model to better define the strategy of EBV infection of human B cells in vivo and to compare this model system with different conditions of EBV infection in humans. Our results support the model of EBV persistence in vivo in cases that were characterized by follicular hyperplasia and a relatively normal CD4⁺ and CD8⁺ T-cell distribution. Intriguingly, in cases that were characterized by nodular and diffuse proliferation with a preponderance of CD8⁺ T cells, similar to infectious mononucleosis, EBV still infects naïve B cells but also induces clonal expansion and ongoing somatic mutations without germinal center reactions. Our results reveal different strategies of EBV infection in B cells that possibly result from variations in the host immune response. Future experiments might allow understanding of the mechanisms responsible for persistent EBV infection and provide targets for more highly tailored therapeutic interventions.     

A rapid hemi-nested PCR for HTLV-I detection.

A hemi-nested PCR approach was adopted to detect HTLV-1 infection in clinical samples of peripheral blood mononuclear cells (PBMCs) from subjects with positive or indeterminate serological results. Our results showed that the hemi-nested PCR quickly solved the diagnostic query, detecting the presence of proviral HTLV-1 DNA in two of the 252 patients with inconclusive serological results. The main advantage of this method are the typology of DNA extraction, allowing a consistent DNA recovery without amplification problems, the rapidity (4-5 hours), the performance of the assay and its comparable or better sensitivity than other HTLV-1 PCR formats.