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Biomedical subjects

M Bologna

Publications and source records attributed to M Bologna.

At least 55 records · Page 3Linked to original sources
PubMed1988

Early diagnosis of prostatic carcinoma based on in vitro culture of viable tumor cells harvested by prostatic massage.

Prostatic cancer is diagnosed too late in most cases, so that therapy is frequently ineffective or even not undertaken at all because of the already advanced stage of the disease. An early diagnosis technique for prostatic cancer would therefore be highly desirable, also because all other available markers give very unsatisfactory results. Because of our experience in tissue culture of human prostatic specimens, by which we have shown good correlations with patient prognosis, we attempted to grow epithelial cells collected from prostatic fluid after rectal prostatic massage. Samples from prostatic cancer patients, diagnosed by needle biopsy, were grown in culture and were able to survive in vitro for at least 2 weeks, thus providing morphological and biochemical data concerning their neoplastic and differentiation features. The early data on this new approach, which we believe might represent a very useful test for the early diagnosis of the neoplasm, are reported here. The method is noninvasive and suitable for mass screening of the disease. Accuracy and reliability of the technique are currently being tested.

Carcinoma↗
PubMed1988

Short-term tissue culture of prostatic carcinoma samples provides useful biological parameters related to patient prognosis.

In order to identify new data to be able to better evaluate patient prognosis in prostatic carcinoma (PRCA), we started a study 6 years ago correlating in vitro parameters from human PRCA samples grown in tissue culture with histological diagnosis of the same tumors [Eur. Urol. 11: 330-333, 1985]. The original study has been extended with more cases and updated with follow-up of the patients. To date, we evaluated 51 specimens of PRCA (18 grade I, 19 grade II, 13 grade III and 1 grade IV) and 8 of benign prostatic hypertrophy. Tissue samples were cultured in medium DME plus 10% fetal calf serum, 10% horse serum and 50 ng/ml each of hydrocortisone and insulin. Epithelial cells grown from the explants showed an average life in culture and morphological and biochemical features in good correlation with tumor grade. Short cultural life span, regular growth and positive secretion activity are typical of low-grade tumors, meanwhile the opposite is true for high-grade tumors. Of these patients, 15 were evaluable for prognosis, because they died or because they were followed-up for at least 3 years. In this group we compared tissue culture data with survival and found a fairly good correlation between growth parameters and clinical outcome of each case. Although more cases are needed to provide statistical significance to the results, the data we collected seem to indicate that low-grade tumors susceptible of poorer prognosis can be identified by a short-term tissue culture showing morphological atypias and long average life span. This method may be easily reproduced in any hospital equipped with a tissue culture unit.

Carcinoma↗
PubMed1988

HPLC reveals famotidine in the urine up to five days after a single 20 mg oral dose.

Using a simple method for the HPLC determination of famotidine (FMTD), a new inhibitor of histamine H2-receptors, it is possible to evaluate the urine levels of the drug in patients undergoing treatment. FMTD is excreted mostly in the urine, in the unmetabolized form. The authors evaluated the endpoint of FMTD levels in the urine in five patients given a single oral dose of 20 mg and found measurable levels of the drug up to 106 h (5 days) after the patients began the medication. This method may be useful for assessing patient compliance in taking the drug: this will allow the gastroenterologist to distinguish patients with true relapses of peptic ulcer disease from false relapses, in clinical trials using this histamine H2-inhibitor.

Administration, Oral↗
PubMed1988

Study of plasma, urine and gastric juice concentrations of famotidine using high performance liquid chromatography.

An improved method for the high performance liquid chromatography (HPLC) determination of famotidine, a recently introduced inhibitor of histamine H2-receptors, has been devised. Plasma, urine and gastric juice concentrations of famotidine have been measured in a series of 46 patients hospitalized for gastrointestinal disorders and for other unrelated pathologies, after a single oral dose of 40 mg. Pharmacokinetic data confirmed previously described trends of distribution and metabolism of famotidine, attaining peak plasma levels 1.5-2.0 h after oral administration and high urine levels, in unmodified form, between two and twelve hours. This simple HPLC method may be adopted for monitoring plasma concentrations of famotidine in patients, for assessing patient compliance in taking the drug by measuring urine levels and for examining the relationship between plasma famotidine concentration and the antisecretory effect.

Chromatography, High Pressure Liquid↗
PubMed1987

Effect of hrTNF alpha on TxB2 release by macrophages.

Peritoneal rat macrophages incubated with human recombinant tumour necrosis factor (hrTNF alpha) released 2-3 times more thromboxane (TxB2) than untreated macrophage controls, a result similar to that obtained by incubating them with endotoxin (ET) or calcium ionophore A23187.

Animals↗
PubMed1986

Mitogen dose-dependent effect of weak pulsed electromagnetic field on lymphocyte blastogenesis.

The effects of pulsed extremely low frequency electromagnetic fields on human peripheral blood lymphocyte mitogenesis induced by phytohaemoagglutinin, concanavalin A or calcium ionophore A23187 were studied. The dependence of the field effect on mitogen concentrations was investigated. Field exposure produced strong inhibition of DNA synthesis when optimal doses of mitogens were used, confirming our previous findings. Opposite effects were observed at suboptimal concentration of mitogens. Experiments performed by exposing cell cultures to the field for short periods indicated that a field application of at least 6 h is needed to influence irreversibly lymphocyte blastogenesis.

Calcimycin↗
PubMed1986

Organization of cytokeratin bundles by desmosomes in rat mammary cells.

In a rat mammary epithelial cell line, LA-7, cytokeratin bundles recognized in immunofluorescence by a monoclonal antibody (24B42) disappear after trypsinization of cultures and are gradually reformed after replating. We have followed the time course of cytokeratin filament reappearance by growing cells in low calcium medium (0.1 mM) which prevents desmosome formation, and then shifting to high calcium (1.8 mM) to start the process. By fixing the cells at various intervals and staining them in immunofluorescence for 24B42 cytokeratin and for desmosomal proteins, we found that cell to cell contact and desmosome formation are prerequisites for keratin filament formation in these cells. EGTA treatment, by disassembling desmosomes, causes the cytokeratin filaments to disappear and the 24B42 protein to pass into a soluble form in this cell line, as ascertained by 100,000 g fractionation and immunoenzymatic assay. Cycloheximide treatment also causes cytokeratin filaments to disappear, indicating that protein synthesis is needed for normal filament maintenance. In another related cell line (106A-10a) and in HeLa cells, trypsinization and EGTA exposure do not cause a complete loss of 24B42 immunofluorescence, although distinct filaments disappear, indicating the presence in these cells of different organizing centers, besides desmosomes, for cytokeratin bundle formation. LA7 cells therefore seem to have a cytokeratin system strictly dependent on the presence of desmosomes, which act as an organizing center for filament assembly. 106A-10a cells (also rich in desmosomes) and HeLa cells (showing instead a reduced number of desmosomes) have a cytokeratin system partially or totally independent from that of desmosomes, with different organizing centers.

Animals↗
PubMed1986

Marker evolution during the development of the rat mammary gland: stem cells identified by markers and the role of myoepithelial cells.

Using monoclonal antibodies and other immunological reagents we have identified characteristic markers for various epithelial cell types within the rat mammary gland. We have followed the evolution of cell types from the emergence of mammary ducts from the epidermis in the fetus to adulthood. Throughout mammary development some cells retain a group of markers which characterize the early stages of development. We have previously suggested that these cells are the stem cells for mammary development. In the adult, these cells are present in end buds and in the myoepithelial layer of ducts. We suggest that the myoepithelial layer, which we propose should be called the "basal" layer, contains several cell types, of which two are pluripotent. It contains the stem cells for mammary development, which also are present in end buds, and a precursor of ductules and alveoli. In the ducts, basal cells are probably also the precursor of luminal cells. We propose a scheme of mammary development.

Age Factors↗
PubMed1986

Abnormal spectrin in hereditary elliptocytosis.

An abnormal alpha subunit of erythrocyte spectrin has been described in hereditary pyropoikilocytosis (HPP), a rare hemolytic anemia characterized by erythrocyte budding and fragmentation. In HPP spectrin, the N terminal domain of the alpha subunit (alpha I T80) shows increased susceptibility to tryptic digestion, resulting in cleavage to a 50,000-d peptide, presumably due to a change in primary structure of the alpha I domain which alters conformation and generates the new cleavage site. The functional result of this conformational alteration is marked impairment of spectrin oligomer formation in vitro, consistent with the established role of alpha I T80 in spectrin self-association. In the present study, we demonstrate an abnormal spectrin alpha subunit in two kindreds with hereditary elliptocytosis (HE) that is qualitatively identical to HPP spectrin. Clinical expression of HE in these families ranges from mild elliptocytosis without hemolysis to severe poikilocytic hemolytic anemia clinically resembling HPP. In all affected individuals, a fraction of alpha I T80 is abnormal, as shown by its cleavage during mild tryptic digestion to the 50 kd peptide described in HPP; the fraction of alpha I T80 affected is directly proportional to the severity of clinical expression of HE. Spectrin oligomer formation is likewise impaired to a degree which correlates with hematologic disease. One of the HE kindreds studied demonstrated polymorphism in the spectrin alpha II domain, previously described as a frequent occurrence in blacks. This family also demonstrates a variant alpha III domain in spectrin that has not previously been described. We conclude that the abnormality in the alpha I domain originally described in HPP spectrin is shared by a subset of patients with HE; the severity of clinical expression, ranging from mild nonhemolytic HE to poikilocytic hemolytic anemia, is related to the fractional quantity of the alpha subunit that is affected.

Adolescent↗
PubMed1985

Human prostatic carcinoma in tissue culture: correlations between histological diagnosis and in vitro parameters.

Prostatic cell biology is still largely unknown so that even the natural history of prostatic carcinoma is unpredictable. In order to correlate new observations with the prognosis of patients with prostatic carcinoma of various grades, we followed up 24 in vitro samples from surgical specimens of prostatic carcinoma. Fragments from 7 grade-I, 10 grade-II, 6 grade-III; and 1 grade-IV tumors were cultivated in Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum, 10% horse serum and 50 ng/ml each of hydrocortisone and insulin. Epithelial cells grown from the explants continued to grow for a maximum of 120 days and their morphology varied from a fairly regular monolayer of polygonal cells to irregular patterns of overlapping growth with many giant multinucleated cells. Although our data need a longer clinical follow-up time and larger numbers to achieve any statistical significance, the present findings seem to indicate rather clearly that a short life span in culture, a regular growth and a positive secretion activity is typical of low-grade tumors and that a longer life span, an irregular growth and a negative secretion in vitro are characteristics of high-grade tumors. A longer clinical follow-up of these patients will be important in the future to indicate whether these findings can be of any real prognostic value.

Aged↗
PubMed1985

Norfloxacin in prostatitis: correlation between HPLC tissue concentrations and clinical results.

Prostatic infections are difficult both to diagnose and to treat and have negative influences on the fertility of the patient. Norfloxacin, a new quinolone antibacterial agent particularly active in urinary tract infections, gave excellent results (400 mg every 12 h for 10 days) in the treatment of chronic-relapsing prostatitis in a group of 20 patients. An 85% success rate was recorded at the end of a follow-up period of 30 days without any undesired effects. This clinical study was paralleled by a pharmacokinetic evaluation of the serum, urine and prostatic tissue concentrations attained by the drug, measured by an HPLC assay on biological fluids and tissue extracts from surgical specimens of 15 patients suffering from benign prostatic hypertrophy and given two 400 mg doses of norfloxacin in the 12 h preceding prostatectomy. The average tissue concentrations (1 microgram/gm), as compared with the MIC90 of the drug for the pathogens sustaining prostatic infections, appear quite sufficient to control the bacterial foci responsible for the chronic-relapsing course of prostatitis and easily account for the clinical success rate.

Administration, Oral↗
PubMed1985

Norfloxacin concentrations in seminal plasma measured by HPLC.

Eighteen healthy volunteers aged 22 to 37 (average 30.44 +/- 4.11 years) seeking fertility evaluation at the Fertility and Sterility Centre of the Medical Clinic of the University of L'Aquila Medical School were studied for norfloxacin concentrations in serum, urine and seminal plasma 14-15 h after taking two 400 mg doses of the drug. The extraction of the biological fluids and the high pressure liquid chromatographic (HPLC) assay were performed following a method described previously (1), with seminal plasma samples processed in the same way as serum samples. The results in urine and serum substantially confirmed the figures already established for norfloxacin concentrations in such fluids. Seminal plasma levels (average 4.78 +/- 2.69 micrograms/ml) were about 5 times higher than the corresponding serum levels (average 0.89 +/- 0.59 microgram/ml). The authors believe that such concentrations, attaining a therapeutic range in the seminal plasma after only two 400 mg doses of norfloxacin, support its use as an effective chemotherapeutic agent in the management of male genital infections that frequently account for subfertility conditions in young patients.

Adult↗
PubMed1984

Dimethylsulphoxide stimulation of dome formation in cultured rat prostate epithelial cells.

Primary cultures of normal rat prostate produce cells with epithelial characteristics that may be useful in the study of cell differentiation and prostate physiology. These epithelial cells, grown in DME plus 10% FCS, 10% HS, and 50 ng/ml each of hydrocortisone and insulin, can be induced to differentiate into dome ( hemicyst ) forming cells by 250 mM DMSO, a molecule that acts as a differentiation inducer in Friend erythroleukemia cells, Madin-Darby canine kidney cells, and in mouse and rat mammary epithelial cells. Dome formation in cell culture is the consequence of transepithelial transport of water and ions, resulting in fluid accumulation between the culture dish and the cell layer. Dome formation in prostate epithelial cells, besides representing an example of induction of differentiation in a cell culture system, suggests that prostate (ductal?) epithelial cells have fluid reabsorption capabilities.

Animals↗