Molecular and functional characterization of polynucleotide phosphorylase from the antarctic eubacterium Pseudoalteromonas haloplanktis.

Polyribonucleotide phosphorilase from the psychrophilic Antarctic eubacterium Pseudoalteromonas haloplanktis (PhPNPase) has been purified. This enzyme catalyzes both the RNA polymerisation and degradation reaction, showing the highest activity at temperatures below 40 degrees C. PhPNPase is quite sensitive to heat treatment and it is endowed with remarkable halotolerance.

Increased metastatic lymph node 64 and CYP17 expression are associated with high stage prostate cancer.

The metastatic lymph node 64 (MLN64), which is localized in the human chromosome 17, encodes a protein with strong homology with steroidogenic acute regulatory protein. Its overexpression in human breast carcinomas and MLNs led to the hypothesis that this protein could be involved in intraneoplastic steroidogenesis. In the present study, we investigated the expression of MLN64 in prostate cancer, another hormone-dependent tumor, and compared its expression with that of CYP17, the gene encoding for the key enzyme of androgen synthesis. We investigated by RT-PCR the expression of MLN64 and CYP17 in 60 prostatic tumors and compared their expression with the stage of disease and the appearance of relapses in a follow-up of 24 months. We found MLN64 and CYP17 expressed in all samples examined, with significantly higher expression in neoplastic tissues with respect to normal tissues (NTs). Moreover, only in neoplastic but not in NTs, a positive linear correlation was found between MLN64 and CYP17 gene expression. MLN64 and CYP17 expression seems to correlate with high stage, high Gleason score and short relapse-free time. These data, for the first time, demonstrate the presence of MLN64 and CYP17 expression in both normal and neoplastic prostatic tissues. The biological role of MLN64 in human prostate and, particularly, in neoplastic tissue is still unclear. Our findings concerning MLN64 and CYP17 gene expression and their significant positive correlation in human prostate cancer may suggest their possible role in intraneoplastic autonomous steroidogenesis.

The IGF axis in prostate cancer.

Prostate cancer, the most frequent non-cutaneous malignancy in men from industrialized countries, is a growing medical problem, representing the second leading cause of male cancer deaths. In the last decade, converging evidence from epidemiological and biological studies suggests that the Insulin-like Growth Factor (IGF) axis is involved in the tumorigenesis and neoplastic growth of prostate cancer. Epidemiological observations indicated that circulating IGF-I levels are positively associated with the increased risk of prostate cancer. The activation of type I IGF receptor (IGF-IR) by IGF-I and/or IGF-II, has mitogenic and antiapoptotic effects on normal and malignant prostate cells. Altered expression of IGF axis components has also been reported in vitro and in animal models of prostate cancer, as well as in human prostate cancer tissue samples. In this review we address and analyze epidemiological studies, in vitro and in vivo cancer models, and human ex vivo prostate cancer researches performed to date supporting the role of IGF axis in prostate cancer.

Long-term persistence with antihypertensive drugs in new patients.

The objective of this study was to investigate stay-on-therapy patterns over 3 years among patients prescribed different classes of antihypertensive drugs for the first time. A retrospective analysis of information recorded in the drugs database of the Local Health Unit of Ravenna (Italy) was carried out on 7312 subjects receiving a first prescription for diuretics, beta-blockers, calcium channel blockers, angiotensin-converting enzyme (ACE) inhibitors or angiotensin II antagonists between 1 January and 31 December 1997. Patients were followed up for 3 years. All prescriptions of antihypertensive drugs filled during the follow-up periods were considered. The patients continuing or discontinuing the initial treatment, the duration of treatment, and the doses taken were all calculated, as well as main factors influencing the persistence rate. The drugs prescribed were predominantly ACE-inhibitors, followed by calcium channel blockers, diuretics, beta-blockers and angiotensin II antagonists. A total of 57.9% of patients continued their initial treatment during the 3-year follow-up period, 34.5% discontinued the treatment, whilst 7.6% were restarted on a treatment in the third year. Persistence with treatment was influenced by: age of patient (persistence rate increasing proportionately with advancing years), type of drug first prescribed (persistence rate higher with angiotensin II antagonists, progressively lower with ACE-inhibitors, beta-blockers, calcium channel blockers and diuretics), gender of patient (persistence was better in males), age of general practitioner (GP) (the younger the GP, the better the persistence rate) and gender of GP (better stay-on-therapy rate with male GP prescribing). In the case of patients treated continuously, mean daily dose increased progressively over the 3 years. With adequate markers, helpful data can be collected from prescription claims databases for the purpose of monitoring the persistence of patients in continuing their medication, and the quality of antihypertensive treatment in a general practice setting.

Pharmacoutilization of antihypertensive drugs: a model of analysis.

OBJECTIVES: To identify clinical and economic indicators of pharmacoutilization of antihypertensive drugs. PATIENTS AND METHODS: 4614 subjects receiving a first prescription for amlodipine, atenolol, fosinopril, indapamide, or losartan were included in the study. All prescriptions filled during the study period from January 1, 1997 to December 31, 1998 were considered. A retrospective analysis was carried out on information recorded in the drug database. The percentage of patients continuing, discontinuing, and switching the initial treatment, duration of treatment, and doses used were calculated together with total costs. RESULTS: A large proportion of patients (65.1%) discontinued the treatment. From the analysis of the mean daily dose taken by patients who continued the treatment, it was found that many subjects took a drug dosage which was below the therapeutic dose range, whereas the administration of doses above the therapeutic range occurred only occasionally. Continuation of treatment accounted for 48.1% of total costs, switching accounted for 20.8%, and discontinuation represented 31.1% of total expenditures. CONCLUSIONS: With adequate markers, helpful data can be collected for monitoring the quality of antihypertensive drug prescriptions and the rational usage of resources in the general practice setting.

Regional variations of insulin-like growth factor I (IGF-I), IGF-II, and receptor type I in benign prostatic hyperplasia tissue and their correlation with intraprostatic androgens.

Benign prostatic hyperplasia (BPH) is an androgen-dependent disease; it originates exclusively in the inner prostate, which includes tissue surrounding the urethra. Stromal-epithelial interaction has a pivotal role in the regulation of the development and growth of the prostate, and locally produced peptide growth factors are considered important mediators of this interaction. Insulin-like growth factor I (IGF-I) and IGF-II, acting mainly through type 1 IGF receptor (IGFR1), have mitogenic and antiapoptotic effects on epithelial and stromal prostatic cells. In this study the expression of IGF-I, IGF-II, and IGFR1 messenger ribonucleic acid (mRNA), the immunoreactive content of IGF-I (irIGF-I) and IGF-II (irIGF-II) were determined in periurethral, intermediate, and subcapsular regions of BPH tissue to verify their possible regional variation; a correlation to the tissue levels of dihydrotestosterone (DHT) and 3 alpha-androstanediol (3 alpha Diol) was also determined to verify their possible androgen dependence. Prostates were removed by suprapubic prostatectomy from 14 BPH patients and sectioned in the periurethral, intermediate, and subcapsular regions. Gene expression of IGF-I, IGF-II, and IGFR1 was evaluated by semiquantitative RT-PCR, using beta-actin as a control. irIGF-I was measured by RIA, and irIGF-II was measured by IRMA after acidification and chromatography on Sep-Pak C(18) cartridges. DHT and 3 alpha Diol concentrations were evaluated by RIA after extraction and purification on Celite microcolumns. IGF-II and IGFR1, but not IGF-I, mRNA was higher in the periurethral than in the intermediate (P < 0.05) and subcapsular (P < 0.01) region. Also, prostatic levels of irIGF-II, expressed as picomoles per g tissue, were higher in the periurethral (20.84 +/- 1.84) than in the intermediate (14.81 +/- 2.11; P < 0.05) and subcapsular (10.88 +/- 1.21; P < 0.001) region. No significant differences were found in irIGF-I content. Considering prostatic androgen levels, DHT and 3alphaDiol presented a regional variation, with the highest concentrations in the periurethral region. IGF-II mRNA and irIGF-II levels were positively correlated with both DHT and 3 alpha Diol content. These results demonstrate that in BPH tissue a greater IGF-II activity is present in the periurethral region, the site of origin of BPH. Moreover, we can hypothesize that the tissue androgen content may modulate prostatic production of IGF-II, acting at the transcriptional and probably the posttranscriptional level. Therefore, even though further studies will need to confirm this hypothesis, DHT may increase IGF-II activity, mainly in the periurethral region, which, in turn, induces, through IGFR1, benign proliferation of both epithelial and stromal cells, characteristic of BPH.

[Epidemiologic model for the assessment of health needs]. / Un modello epidemiologico per la valutazione dei bisogni di salute.

Health is a high priority in public health planning, a process by which health, services and resources are connected to each other. Assessing peoples health needs is one of the most difficult tasks, because of the lack of information on the spread of diseases, the seriousness of them and their capacity to disable, as well as the lack of timely and appropriate information on them. The epidemiological model put forward describes the volume of resources necessary to maintain the health needs of a population as depending upon the prevalence of diseases, the health services used to combat these diseases and the consequent human and financial resources employed. According to this model the management of the supply of health services depends on monitoring the relationship between population, disease, health services and resources.

Hyperandrogenism in the adolescent female.

Hirsutism in adolescent girls commonly starts as an esthetic problem in young women and is later complicated by the development of infertility and polycystic ovary syndrome, which are frequent consequences of prolonged hyperandrogenism. To ascertain whether particular prepubertal clinical manifestations may predict the development of adolescent hirsutism, we followed 70 girls with precocious pubarche (PP) with or without prepubertal hypertrichosis (PH) until 3 years (mean age 14.8 +/- 0.9 years) after menarche. Similar follow-up was carried out in six girls with PP secondary to 21 hydroxylase deficiency (NC-CAH), treated with hydrocortisone. In addition, a retrospective study on the incidence of precocious pubarche was performed in 139 hirsute teenagers (mean age 17 +/- 1.8 years). Testosterone, androstenedione, dehydroepiandrosterone sulphate, 17 alpha-hydroxyprogesterone (basal and after ACTH), luteinizing hormone and follicle-stimulating hormone were evaluated by radioimmunoassay or immunoradio metric assay in the early follicular phase, in cycling subjects. Pelvic ultrasonography was also performed. In the 139 hirsute teenagers, 29 had a history of PP (21% vs. 0.6% in the general Italian population). Of these 139 patients, NC-CAH was diagnosed in 8 (6%), 5 of whom (63%) had PP. Of the 70 girls with PP, hirsutism was present in 44 (63%). PH was present in 37 of 44 patients (84%) with hirsutism, but only in 9 of 26 (35%) without hirsutism. Our results showed that 1) PP represents a risk factor for the development of postpubertal hirsutism; 2) the association with PH seems to increase the risk probability; and 3) patients with hirsutism due to NC-CAH have a higher incidence of PP compared with other hirsute patients, but glucocorticoid treatment in such patients prevents the development of hirsutism. Whether early treatment in the other PP patients may prevent the development of hirsutism remains to be established.

Synergistic transcriptional activation of the thyrotropin receptor promoter by cyclic AMP-responsive-element-binding protein and thyroid transcription factor 1.

In this study we have investigated the molecular mechanisms involved in hormonal induction of thyroid-specific transcription of the thyrotropin receptor (TSHr). A cyclic AMP-responsive element (CRE) has been characterized in the minimal TSHr promoter, and promoter activity shown to be also induced by thyroid transcription factor 1 (TTF-1). We here describe a cooperative effect between TTF-1 and CRE-binding protein on the TSHr promoter. Moreover we have identified a second TTF-1-binding site in the minimal promoter, which does not activate TSHr promoter activity but is required for the co-operative activation of the promoter. This report describes a new aspect of thyroid-specific gene expression, namely, how a generic extracellular signal can be interpreted in a thyroid-specific way.

[SO2 air pollution and hospital admissions in Ravenna: a case-control study]. / Inquinamento atmosferico da SO2 e ricoveri ospedalieri a Ravenna: un'analisi caso-controllo.

The association between air pollution and hospital admission for chronic respiratory and ischemic heart diseases was analyzed through a case-control approach. The study was based on routinely available data in Ravenna and covered the period 1986-88. A control group (n = 37250) was used to evaluate the specificity of the health effects of air pollution. It included all patients admitted into the hospital for conditions other than respiratory or heart diseases. The adjustment for confounding variables (climatic factors and time variables) was attained by multiple logistic regression. The risk of hospital admission for chronic bronchitis (n = 587) increased of 10% for a 10 ppb increase in SO2 concentration (in the range 0-60 ppb). A similar positive association (OR = 1.2) was observed for exposures to more than 10 ppb of SO2 and hospitalization for ischemic heart diseases (n = 1982). An apparently inverse relationship was found between SO2 pollution and hospitalization for asthma (n = 203). The case-control method can be a useful alternative tool to elucidate specific associations, in comparison with the statistical methods most frequently used in time-correlation analysis.

Purification and characterization of thyroid transcription factor 2.

Thyroid transcription factor 2 binds to the promoters of both thyroglobulin and thyroperoxidase genes, two markers of thyroid tissue differentiation, and its binding modulates the activity of both promoters. In this paper we describe the purification of thyroid transcription factor 2 essentially to homogeneity and demonstrate that it is a thyroid-specific DNA-binding protein. Furthermore, we provide a biochemical characterization suggesting that thyroid transcription factor 2 binds to DNA as a dimer and that it is a zinc-finger DNA-binding protein regulated in vitro by the redox state.

The thyroid hormone inhibits the thyrotropin receptor promoter activity: evidence for a short loop regulation.

Thyrotropin, by binding its specific receptor on the plasma membrane of the thyrocyte, regulates thyroid function and differentiation. In FRTL5 cells, thyrotropin down-regulates the thyrotropin receptor (TSHr) promoter activity and induces the transcription of the alpha form of thyroid hormone receptor (TR-alpha 1). In this study we show that the thyrotropin receptor down-regulation, induced by thyrotropin, is mediated by TR-alpha 1. The thyroid hormone receptor binds, in vitro, the thyrotropin receptor minimal promoter and inhibits promoter activity in cotransfections experiments in CV 1 cells. The inhibition is achieved only in the presence of the thyroid hormone. The TSHr promoter mutated in the thyroid hormone receptor binding site does not bind TR, in vitro, and its activity is not inhibited, in cotransfection experiments, in CV 1 cells. The same mutation abolishes the TSH mediated down-regulation of the TSHr promoter activity in FRTL5 thyroid cells. These results support the hypothesis of a regulatory short loop of thyroid hormone in thyroid cells.

Thyroid transcription factor 1 activates the promoter of the thyrotropin receptor gene.

The TSH receptor (TSHr) is one of the most important thyroid differentiation markers. The binding of the TSH hormone to its receptor is an essential step in the modulation of thyroid function and differentiation. Here we report that the thyroid transcription factor 1 (TTF1), a transcription factor essential for thyroid-specific gene expression, binds to the TSHr minimal promoter. The promoter, when mutated at this binding site, shows a decreased activity in thyroid cells. In cotransfection experiments in nonthyroid cells, TTF1 is able to trans-activate the TSHr minimal promoter. This finding strengthens the importance of TTF1 in the maintenance of thyroid differentiation. The promoters of the main thyroid differentiation markers thyroglobulin, thyroperoxidase, and now TSHr, are regulated by TTF1.

Decline of incidence of A, B and non-A, non-B hepatitis in Italy. Results of four years surveillance (1985-88). SEIEVA collaborating group.

A specific surveillance system for acute viral hepatitis which uses weekly notification of cases and a standard risk factor questionnaire was started in Italy in 1984. From 1985 to 1988 153 Health Departments (27% of Italian population) joined the system. Estimated incidence of reported viral hepatitis was 30 per 100,000 in 1985 and 14 in 1988. The decline was impressive for hepatitis A, particularly in the south. Reduction of incidence was also evident for hepatitis B and Non-A, Non-B hepatitis, particularly in young adults. The decrease of viral hepatitis in Italy is consistent with recent seroepidemiological data. Shellfish consumption was the most frequent risk factor reported for hepatitis A cases at all ages. Hospitalization, surgical intervention, dental therapy and other percutaneous exposures still play a role in the transmission of parenteral hepatitis in Italy. Blood transfusions seem to be important only for Non-A, Non-B. Interventions other than vaccination to prevent B and Non-A, Non-B hepatitis due to hospitalization, surgical intervention, other percutaneous exposures and dental therapy are needed and can further contribute to the decline of hepatitis virus infection rates in Italy.

The role of antiseptics and disinfectants in the control of nosocomial infections.

More than 600,000 cases per year of nosocomial infection in Italy reflect, in part, the scant attention given to the vehicles of infection, conditions which favour cross-contamination and not least to the inappropriate use of antiseptics and disinfectants in our hospitals. Disinfection procedures are intended to remove or neutralise sources of potentially pathogenic organisms or to prevent their transmission by blocking the routes of transmission. Data to support many disinfection policies, particularly those of routine disinfection of inanimate surfaces, i.e. floors and walls, appear unable to show significant benefit to the patient. With the exception of particular areas, a correlation between reduction in counts of airborne bacteria and microbial colonisation or clinical infection has failed to be demonstrated. However, certain antiseptic practices e.g. correct hand washing procedures, surgical skin preparation and instrument sterilisation are of invaluable use in the control of nosocomial infection. It is therefore of critical importance in this period of cost cutting and financial restraint within the hospital services that we adopt those infection control measures which are of greatest cost effectiveness. A recent critical evaluation of the use of antiseptics in several Italian hospitals, illustrates the principal characteristics of antiseptics and their fundamental role in the control of hospital infection. This study also highlights common misuses of antiseptics and their cost/benefit ratio.