The product of the retinoblastoma-related human gene Rb2/p130 is highly homologous with the product of the retinoblastoma tumor suppressor gene (pRb) and Rb-related p107. this homology is shared mainly in the pocket domain, a region that seems to play a key role in the functions of these proteins. Here we report the molecular cloning and initial characterization of the cDNA encoding the murine homologue of the human Rb2/p130 gene product. The 4.8-kb cDNA encodes a protein of 1125 amino acids that shows 90% indentity to that of the human protein. The Rb2/p130 mRNA is found to be expressed in all of the adult mouse tissues examined, with the highest level being detected in kidney and skeletal muscle. For the protein characterization, we used a polyclonal antibody raised against the COOH terminus of the human Rb2/p130 protein that also recognizes the mouse protein. In developing mouse embryos, the Rb2/p130 protein is expressed as early as day 10 of gestation and reached a peak of expression around day 13 of gestation, implying a developmental regulation of the Rb2/p130 gene in murine ontogeny.

3T3 Cells↗

PubMed1994

p130/pRb2 has growth suppressive properties similar to yet distinctive from those of retinoblastoma family members pRb and p107.

The retinoblastoma tumor suppressor gene product, as well as its related protein p107, has been shown clearly to exert its growth suppressive effects in a cell cycle dependent manner. In this study we demonstrate that the introduction of our recently cloned Rb family member p130/pRb2 causes growth arrest in three tumor cell lines. In addition, in the nasopharyngeal carcinoma derived cell line HONE-1, we identified a low level of expression of p130/pRb2, possibly due to gene rearrangement, and a drastic reduction in proliferation upon introduction of a constitutive active p130/pRb2 complementary DNA clone. Furthermore, we were able to dissect distinct properties of the Rb family by demonstrating that p130/pRb2 inhibits proliferation of the glioblastoma cell line T98G, which is resistant to the growth suppressive effects of both pRb and p107. Our studies demonstrate that the Rb family proteins identified to date may complement each other but they are not fully functionally redundant.

Cell Division↗

PubMed1994

Monoclonal antibodies against human breast tumor-associated antigens: characterization of B21 antigen.

BACKGROUND: Monoclonal antibodies (MAbs) show promise in the early detection and monitoring of cancer and may have therapeutic applications as well. PURPOSE: The purpose of this study was to characterize MAb B21, a novel murine-derived antibody that has strong reactivity with MCF-7 and T47D cell lines from human breast cancer. METHODS: A number of MAbs that react with breast cancer cell lines were obtained from cultured mouse spleen cells, and one, MAb B21, was selected for detailed analysis. MAb B21 was characterized by immunocytochemical, immunofluorescence, immunoprecipitation, and Western blotting procedures. RESULTS: We found a strong reactivity of MAb B21 with cultured breast cancer cells and cells from human breast tumors, although some reactivity was seen sporadically in non-breast or normal tissue. Negligible reactivity was detected in a series of non-breast cell lines and with normal breast epithelial cell line MCF-10A. However, when MCF-10A cells were permeabilized, allowing the antibody to penetrate within the cells, reaction became apparent. MCF-10A cells, when transfected with the oncogene c-Ha-ras (MCF-10T), gave a positive immunostaining similar to that observed with MCF-7 and T47D cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of L-[35S]methionine-labeled MCF-7 and T47D cell extracts showed distinct immunoprecipitated components, with molecular weight values ranging from 150,000 to 20,000 with the addition of MAb B21. Western blot assays using MAb B21 of SDS-PAGE fractionated/electroblotted proteins from breast cancer cell lines and MCF-10A cells showed specific reaction with a 95,000 component. CONCLUSIONS: Our results indicate that B21 antigen is expressed in neoplastic cells of epithelial origin, mainly breast cancer, and to a minor extent in other cell lines. In addition, MAb B21 recognizes an antigen that is differentially localized during cell transformation. IMPLICATIONS: Our future studies will address the full characterization of MAb B21 and explore its capacity as a tool for therapeutic manipulation.

Antibodies, Monoclonal↗

PubMed1994

Hormonal and metabolic responses to the stress of transport and slaughterhouse procedures in clenbuterol-fed pigs.

The aim of this study was to evaluate if metabolic and endocrine adaptive responses to stressful stimuli are influenced by clenbuterol treatment in pigs. Two groups of nine Goland barrows of 167 kg liveweight, fed diets containing 0 (control) or 1 ppm of clenbuterol (clenbuterol-treated) for 40 days, were used. Blood samples were collected by jugular venipuncture the day before (day 40) and at slaughter (day 41). On day 40 clenbuterol-treated pigs had lower levels of thyroxin, tri-iodothyronine, insulin, urea and higher levels of lactate and creatine phosphokinase activity. On day 41, thyroxin concentration was lower and creatine phosphokinase activity higher in clenbuterol-treated pigs than in controls. In response to transport and slaughterhouse procedures, most of the parameters considered both in clenbuterol-treated and in control pigs increased.

Animals↗

PubMed1994

Epidermal growth factor in the topical treatment of herpetic corneal ulcers.

The tolerability and efficacy of epidermal growth factor (EGF) in the topical treatment of herpetic corneal ulcers in addition to topical acyclovir have been evaluated in a double-blind, placebo-controlled, randomized study in two groups of patients. The time required for complete reepithelialization of the cornea was recorded, and the data obtained were analyzed statistically. In the EGF group, the reepithelialization was significantly faster than in the control group. Tolerability of EGF was always excellent. These results indicate that EGF is safe and effective in reducing the healing time of herpetic corneal ulcers.

Acyclovir↗

PubMed1993

Vacuolar apical compartment (VAC) in breast carcinoma cell lines (MCF-7 and T47D): failure of the cell-cell regulated exocytosis mechanism of apical membrane.

We have previously shown that an integral plasma membrane glycoprotein (AP2) is highly polarized to the apical domain in confluent Madin-Darby canine kidney (MDCK) epithelial cells. However, when the monolayers are prevented from forming intercellular contacts, approximately 60% of the AP2 cellular content is stored in the intracellular vacuolar apical compartment (VAC). In the current work we found that AP2 was present in the non-tumorigenic human mammary epithelial cell line MCF-10A, in the breast carcinoma cell lines MCF-7 and T47D, and in breast ductal carcinomas in vivo. By radioimmunoassay, an intracellular compartment of AP2 was identified in the mammary cell lines in culture. In MCF-10A, this compartment behaved as in MDCK cells; namely it was observed only when the cells cannot form cell-cell contacts. However, in the carcinoma cell lines MCF-7 and T47D, a significant AP2 intracellular compartment was observed also under conditions permissive for the formation of intercellular contacts. These results were confirmed by immunofluorescence and immunoelectron microscopy experiments that showed VACs in MCF-7 and T47D, even in cells with extensive intercellular contacts. In MCF-7 cells, the addition of serum caused a partial decrease of the AP2 intracellular compartment. The exocytosis of VACs occurred towards the center of multi-cellular groups, forming intercellular lumens, similar to those transiently observed in MDCK cells and to structures described by others during embryo development. Altogether, these results suggest that VAC exocytosis is controlled by cell-cell contact signalling, which may be defective in carcinoma cells.

Animals↗

PubMed1993

Autologous bone marrow purging with LAK cells.

In this study we will demonstrate that LAK cells, in vitro, can lyse hematologic neoplastic cells with a minor toxicity of the staminal autologous marrow cells. In fact, after bone marrow and LAK co-culture at a ratio of 1/1 for 8 hours, the inhibition on the GEMM colonies resulted to be 20% less compared to the untreated marrow. These data made LAK an inviting agent for marrow purging in autologous bone marrow transplantation.

Bone Marrow Purging↗

PubMed1993

The retinoblastoma-related gene, RB2, maps to human chromosome 16q12 and rat chromosome 19.

A retinoblastoma-related human gene, referred to as RB2, has been cloned based on sequence homology of the E1A-binding domain of the retinoblastoma gene. Structural homology with the retinoblastoma gene suggests a possible function of RB2 as a tumor suppressor gene. In this study, we have mapped this gene to human chromosome 16q12.2 and rat chromosome 19, using fluorescence in situ hybridization and somatic hybrid cell analysis, respectively. Based on known syntenic relationships among human, rat and mouse, the data suggest that the mouse homolog resides on chromosome 8. Deletions of chromosome 16q have been found in several human neoplasias (including breast, ovarian, hepatic, and prostatic cancers) which is in support of an involvement of RB2 in human cancer as a tumor suppressor gene.

Animals↗

PubMed1993

Cloning of a new member of the retinoblastoma gene family (pRb2) which binds to the E1A transforming domain.

The product of the retinoblastoma tumor suppressor gene (pRb) and p107 share a high degree of structural homology in the pocket region, which is thought to play a primary role in the function of these proteins. It is conceivable that there exists a larger family of cellular proteins containing this pocket region. In this communication, we report cloning of a new human cDNA encoding a polypeptide that shows a high level of identity with pRb and p107 and possesses a pocket region. We have named it pRb2. From the deduced amino acid sequence, pRb2 has a predicted molecular weight of approximately 120 kD and its in vitro translated product binds to the adenovirus E1A protein. Due to its size, pRb2 may correspond to p130, which has previously been shown by us to interact with the transforming region of E1A in in vivo studies. Interestingly, pRb2 fails to bind an E1A mutant in the transforming domain 2 suggesting that pRb2 is involved in the transforming capacity of E1A.

Adenovirus E1A Proteins↗

PubMed1991

Vitreous fluorophotometric recordings in HIV infection.

Fluorophotometry was performed in 10 patients. Five of these were affected by AIDS and five ARC (AIDS Related Complex). All patients had normal fundi detectable by standard examination and by fluorescein angiography. In spite of the normal condition of the fundus, vitreous fluorophotometry readings were constantly above normal. We believe that the disturbance of the blood retinal barrier appears before any lesion detectable both clinically and by fluorangiography. It is possible that the first toxic effect of the retrovirus is at the level of the retinal vessels.

AIDS-Related Complex↗

PubMed1991

Bioelectrical impedance assay (BIA) of total body composition in alcohol-induced migraine patients. Preliminary report.

"Whole body composition" was investigated in 30 people subject to alcohol-induced migraine. The control group was 30 people matched for age, sex, height and weight, who were not subject to migraine and who consumed moderate amounts of alcohol. The following were evaluated: anthropometric data, alcohol intake in grams per day, total body water, fat-free mass, fat mass, and body mass index (kg/m2). Two methods of measurement were used: skin fold thickness evaluation, and bioelectric impedance assay (BIA). BIA is a non-invasive method based on the principle that lean tissues conduct a low frequency alternating current of electricity better than do fatty tissues. Body mass index was increased in the alcohol-induced migraine patients (p less than 0.04), as was fat mass, (p less than 0.03) as evaluated by skin fold measurements. Bioelectric impedance assay demonstrated a slight total body water increase in alcohol-induced migraine patients (p less than 0.07). Possibly, if this total body water increase was reflected at the neuronal level, the bioelectric properties of the migrainous brain may be altered, thus creating a hypothetical link with the phenomenon of spreading depression.

Adolescent↗

PubMed1990

Fine needle aspiration biopsy of the prostate gland: our experience concerning 101 cases with histological follow-up.

Our experience concerning 605 fine needle aspiration (FNA) biopsies performed between 1985 and 1988 is reported. FNA specimens of the prostate gland were compared to histological material in 101 cases: 37 patients underwent suprapubic prostatectomy, 15 radical prostatectomy, 28 transurethral resection, and 21 core needle biopsies. Adenocarcinoma was correctly diagnosed by using cytology in 39 out of 40 cases; benign prostatic hypertrophy was confirmed by histology in 54 out of 57 cytologically benign cases. The absolute sensitivity of the FNA biopsy was 98.2%; specificity was 98.1%; efficiency was 96%; and false negative rate was 6.6%. Our data support the value of transrectal aspiration biopsy as a precise and easy method for diagnosing prostatic cancer; the low false negative rate and the high number of correct diagnoses underline the great accuracy of the method.

Adenocarcinoma↗

PubMed1989

[Total parenteral nutrition. Analysis of the results of 144 treated cases].

Of 144 patients treated with T.P.N. in the period 1-5-1987/30-9-1988, 128 have been to have an operation. Their survival at the time of the removal, the post-operating mortality, the average stay in hospital, have been compared with those of other 128 patients non-treated with T.P.N., suffering from similar pathologies and who had operation of the same kind. The obtained results have stressed in the patients treated with T.P.N. as in those who had not, a survival at the time of the removal of 75% contrary to 62.5%, a post-operating mortality of 25% contrary to 37.5%, an average post-operative stay in hospital of 13 days contrary to 15 days. The statistical elaboration executed with chi 2's test with Yates' correction, provided 0.25 less than P greater than 0.50, so data non particularly meaningful. In our casuistry the percentage of surviving in the patients treated with T.P.N. as to those not treated, is nevertheless, very encouraging; furthermore, we most think that only by taking advantage of T.P.N. has been possible to extend the operability to patients that in the past because of well on years, or because of dangerous surgical illnesses joined to dismetabolic and cardiovascular diseases, were judged not operable.

Aged↗

PubMed1989

Effects of blood sampling procedures, grouping and adrenal stimulation on stress responses in the growing pig.

Changes in the blood and the behaviour of 14 growing pigs from 4 different litters were evaluated under different experimental conditions of blood sampling, grouping and adrenal stimulation. The results showed that the different techniques of blood sampling influenced lactic dehydrogenase (LDH) and creatine kinase (CK) activities. Cortisol, proteins and CK levels were negatively correlated with social hierarchy after regrouping. Cortisol was also correlated with total activity levels. Adrenal stimulation by adrenocorticotropic hormone (ACTH) administration caused a sharp increase in plasma cortisol levels. However, plasma glucose, plasma proteins and total leukocyte counts were not affected by the ACTH treatment.

Adrenal Cortex↗