External quality assessment for the detection of HCV RNA, HIV RNA and HBV DNA in plasma by nucleic acid amplification technology: a novel approach.

BACKGROUND AND OBJECTIVES: In this EQA study a novel approach was used to assess the performance of blood centres and blood product manufacturers in detecting the possible contamination of plasma with HCV, HIV and HBV by NAT. MATERIALS AND METHODS: A panel of 12 samples, three negative and three positive for each virus, was distributed to the EQA participants. The positive samples were prepared, using the respective WHO standards, in order to obtain a viral concentration of about three times the 95% DL of the methods most commonly used by laboratories involved in blood screening by NAT. Participants were requested to test each sample of the panel on different days, possibly by different operators using their routine NAT assay. RESULTS: Overall, the participants' performance was satisfactory. In particular, 49 of the 59 participants (83%) were able to correctly identify all samples. Regarding the remaining 10 laboratories, in three cases a deviation from the laboratory's procedure that could be attributed to an operator's mistake was observed, in two cases a possible cross-contamination occurred while in the remaining five cases the failure to detect the positive samples couldn't be ascribed to any relevant deviation in the laboratory's procedure. CONCLUSIONS: The novel design of this EQA study allowed participants to verify their day by day activity as the study was carried out in the context of their routine testing. Under these conditions, it was demonstrated that, despite the high level of automation reached by NAT assays, human errors can still occur.

Brush-border-enzyme-mediated intestine-specific drug delivery. Amino acid prodrugs of 5-aminosalicylic acid.

5-Aminosalicylic acid (5-ASA) is the active principle of a number of preparations aimed at the treatment of inflammatory bowel diseases, such as Crohn's disease and ulcerative colitis, but its efficacy is limited by early absorption and metabolism. The possibility to exploit the selective hydrolytic activity of brush border enzymes such as aminopeptidase A and carboxypeptidases was studied by preparing the following four amino acid prodrugs of 5-ASA: 5-(N-L-aspartylamino)-2-salicylic acid, disodium salt (18), 5-(N-L-glutamylamino)-2-salicylic acid, disodium salt (19), [(5-aminosalicyl)-L-prolyl]-L-leucine, sodium salt (25), and [[5-(N-L-glutamylamino)salicyl]-L-prolyl]-L-leucine, disodium salt (28). In these compounds, the peptide bond is selectively split by the intestinal brush border aminopeptidase A (compounds 18, 19, and 28) and carboxypeptidases (compounds 25 and 28).

Granulomatous slack skin.

A 39-year-old man had a two-year history of three indurated plaques that progressively underwent a cutis laxa-like transformation. Histopathologically a granulomatous, deep, dermal infiltrate was observed. It was composed mainly of numerous histiocytes and many multinucleate cells and foam cells. The case has the clinical and microscopic features of granulomatous slack skin (GSS), which may be considered to be a peculiar form of localized elastolysis. To date, there is no effective therapy for GSS.

Contribution of GABAergic cortical circuitry in shaping somatosensory evoked scalp responses: specific changes after single-dose administration of tiagabine.

OBJECTIVES: To determine whether conventional as well as high-frequency somatosensory evoked potentials (SEPs) to upper limb stimulation are influenced by GABAergic intracortical circuitry. METHODS: We recorded SEPs from 6 healthy volunteers before and after a single-oral administration of tiagabine. Conventional low-frequency SEPs have been obtained after stimulation of the median nerve, as well as after stimulation of the first phalanx of the thumb, which selectively involves cutaneous finger inputs. Median nerve SEPs have been further analyzed after digital narrow-bandpass filtering, to selectively examine high-frequency responses. Lastly, in order to explain scalp SEP distribution before and after tiagabine administration, we performed the brain electrical source analysis (BESA) of raw data. RESULTS: After tiagabine administration, conventional scalp SEPs showed a significant amplitude increase of parietal P24, frontal N24 and central P22 components. Similarly, BESA showed a significant strength increase of the second peak of activation of the first two perirolandic dipoles, which are likely to correspond to the N24/P24 and P22 generators. By contrast, no significant changes of high-frequency SEPs were induced by drug intake. CONCLUSIONS: Our findings support the view that both N24/P24 and P22 SEP components are probably generated by deep spiny cell hyperpolarization, which is strongly increased by inhibitory inputs from GABAergic interneurons. By considering the clear influence of inhibitory circuitry in shaping these SEP components, conventional scalp SEP recording could be useful in the functional assessment of the somatosensory cortex in different physiological and pathological conditions. By contrast, intrinsic firing properties of the cell population generating high-frequency SEP responses are unaffected by the increase of recurrent GABAergic inhibition.

Ornithine decarboxylase in colonic mucosa from patients with moderate or severe Crohn's disease and ulcerative colitis.

Polyamines, as well as ornithine decarboxylase (ODC), the enzyme involved in their synthesis, were reported to be closely related to cell proliferation. In Crohn's disease and ulcerative colitis, cell destruction and proliferation increase in the active stage. The aim of the present study was to determine the ODC in both involved and uninvolved areas of the colonic mucosa of active Crohn's disease and ulcerative colitis patients. The patients were divided in two groups, owing to the different level of activity (severe or moderate), by means of clinical endoscopy, laboratory, and histology evaluations. Subjects with suspected disease, but endoscopically unconfirmed, were used as controls. In all ulcerative colitis and Crohn's disease patients the ODC values both in involved and uninvolved mucosa were significantly lower than in controls. In severe Crohn's disease ODC was significantly reduced versus moderate Crohn's disease only in affected tissues. In all ulcerative colitis patients (moderate or severe) the ODC was significantly decreased in involved mucosa compared with uninvolved mucosa. Severe ulcerative colitis showed the significantly lowest ODC. We suggest that the significant decrease of ODC in the bowel mucosa is closely related to the severity of the disease. The highest decrease of ODC in ulcerative colitis patients would be due both to the enhanced cell destruction, and to the feed-back from exogenous increased polyamine production (bowel bacteria, cell desquamation). Therefore ODC would be considered a sensitive index of the inflammatory derangement of the mucosa, especially in acute ulcerative colitis. We conclude that this behaviour may result in an enhanced risk of neoplasia.

Serum IL-10 levels in HIV-positive subjects: correlation with CDC stages.

HIV-1 infection and the HIV gp120 have been shown to induce an IL-10 increase in cultured peripheral blood mononuclear cells. Furthermore, the expression of this cytokine has been reported to increase in lymphnodes of infected patients along the disease course, and a shift from the TH-1 towards the TH-0/TH-2 phenotypes (with subsequent IL-10 release) has been hypothesized to underly AIDS progression. In this study the serum IL-10 levels found in 30 HIV-negative controls and in 65 HIV-positive patients, untreated with AZT and negative for HBsAg and HCV-Ab have been compared, using a commercial, competitive ELISA method based on a polyclonal anti-IL-10 serum. With this test, HIV-positive sea showed IL-10 levels significantly higher than those found in the controls. In addition the IL-10 levels progressively increased in the subsequent CDC stages, without further changes from the stage III to the stage IV. Accordingly, patients evaluated two times in CDC stage II, with a time interval of at least one year, showed significant IL-10 increases, even more pronounced when the same patients passed from CDC stage II to stage III. Furthermore, a significant, negative correlation was observed between the circulating IL-10 levels and the patients' CD4/CD8 ratios. These data may be important from a clinical point of view since IL-10 monitoring could be considered as a surrogate marker for evaluating the disease progression. In addition, several immunological abnormalities present in HIV positive patients, such as the monocyte/macrophage impairment and the hypergammaglobulinemia could be related to the enhanced IL-10 expression.

Behaviour of several 'progression markers' during the HIV-Ab seroconversion period. Comparison with later stages.

Two acute phase reactants, four cytokines, five soluble factors and lymphocyte subpopulations have been simultaneously evaluated in 16 subjects before and closely after the HIV-Ab seroconversion time. The same variables have also been determined in 50 HIV-Ab-negative high risk subjects, in 36 CDC II-III and in 30 CDC IV patients, utilizing a mixed longitudinal epidemiological model. The results show significant variations of few parameters in the early phases (increase: sCD8, beta-2-Microglobulin, sIL-2R, sCD23, Neopterin, IFN-alpha; decrease: CD4+ lymphocytes). In the course of the disease, many others parameters progressively increase (IFN-tau, IL-4, IL-6, acid-alpha 1-glycoprotein, alpha 1-antitrypsin) or decrease (B- and T-lymphocytes). Ferritin, in particular, highly increases only in CDC IV stage. These data may be useful to monitor patients during the entire course of their disease and to suggest the time elapsed from seroconversion.

Electromagnetic and thermal models of a water-cooled dipole radiating in a biological tissue.

An insulated, water-cooled dipole, radiating in a biological tissue, is analyzed with a theoretical electromagnetic and thermal model. The SAR and temperature distributions are calculated taking into account the effect of the water flowing inside the applicator. The steady-state temperatures in a dissipative medium, interacting with the dipole, are evaluated for several thicknesses of the external casing, water temperatures and blood perfusions. A correct design of the external casing thickness and a proper choice of the temperature and flow velocity of water allows to control the wall temperature of the applicator within physiological limits. The influence of the blood perfusion on the temperature distribution is investigated.

FDTD electromagnetic and thermal analysis of interstitial hyperthermic applicators. Finite-difference time-domain.

In this paper, the electromagnetic and thermal behavior of interstitial applicators was analyzed by using the Finite-Difference Time-Domain method. Two configurations were considered: a simple insulated dipole antenna radiating in a layered tissue, and an air cooled applicator radiating in a tissue-equivalent phantom. The proposed approach allows a detailed modeling of the complete structure of the applicator. Furthermore, Specific Absorption Rate and temperature distributions can be determined considering real clinical or experimental conditions. The temperature distribution for the air cooled applicator has been compared with experimental results.

A water-cooled EM applicator radiating in a phantom equivalent tissue--experiments and numerical analysis.

A prototype of a water-cooled electromagnetic applicator for intracavitary hyperthermia has been tested. The temperature distributions produced in a polyacrylamide dissipative medium have been shown using liquid crystals. A complete electromagnetic and heat transfer model can predict the experimental temperatures.

Risk factors for hepatitis B virus infection among homosexual men attending a sexually transmitted diseases clinic in Italy.

Prevalence of and risk factors for hepatitis B virus (HBV) infection were determined among 252 homosexual men with no history of intravenous drug use (median age 33 years, range 18-77) treated at a sexually transmitted disease (STD) clinic in Rome. The overall prevalence of antibodies to HBV core antigen (anti-HBc) was 50.8%, a rate nearly nine times as high as the 5.8% found recently in a national sample of young male adults, aged 18-26 years, and twice as high as the 22% found in heterosexuals attending the same clinic over the same period of time. Multiple logistic regression analysis showed that the risk of anti-HBc positivity was independently associated with increasing age, five or more sexual partners in the previous year, positive HIV serology and positive syphilis serology. Lower level of schooling, lack of condom use, history of non-ulcerative STD, current or past history of genital herpes, and positive anti-HCV serology were not associated with anti-HBc positivity. These findings corroborate the importance of sexual transmission of HBV in homosexual men. Behavioural factors, such as multiple sexual partners, probably enhance the efficiency of this mode of HBV transmission.

Interferon plus ursodeoxycholic acid versus interferon in the treatment of chronic C viral hepatitis.

BACKGROUND: The aim of as study was to ascertain whether the association of interferon alpha-2a and ursodeoxycholic acid (IFN+UDCA) was more efficacious in ameliorating liver parameters than interferon (IFN) alone in patients with chronic hepatitis C. METHODS: Forty-one chronic hepatitis C patients, who had at least twice the normal value of one transaminase, were randomly assigned to treatment with IFN + UDCA (n = 21) or IFN alone (n = 20). IFN was administered subcutaneously at a dose of 3 MU thrice weekly, UDCA orally at 10 mg/kg bw/day. IFN therapy was terminated 6 months later and the responders (normalized transaminases) of both groups were treated with UDCA alone for a further 12 months. RESULTS: In the IFN + UDCA group there were 2 drop-outs from therapy and 11 responders, while in the IFN group they were, respectively, 3 and 10. Transaminases normalized after the first month of treatment in 7/11 responders with IFN + UDCA compared with 3/10 in the IFN responders group. The trend to normalization was more rapid with IFN + UDCA than with IFN alone (chi 2t = 3.95; p < 0.05). Disease relapse (defined as at least one transaminase > x 1.5 the normal value) was 3/11 in the IFN + UDCA group and 4/10 in the IFN group. 2/11 responders in the IFN + UDCA and 1/10 in the IFN group were HCV RNA negative by PCR. The total Knodell histological score decreased more in the IFN+UDCA than in the IFN group (-2.67 +/- 3.44. vs -1.67 +/- 2.16, mean +/- SD). CONCLUSIONS: The administration of UDCA determine an earlier normalization-time of transaminases in the patients responders to IFN therapy and could be useful to reduce the relapse into disease after the IFN therapy.

[Qualitative and quantitative significance of the microbial flora in natural mineral waters during their bottling]. / Significato qualitativo e quantitativo della flora microbica nelle acque minerali naturali durante la loro commercializzazione

On the basis of data reported in literature concerning the saprophytic bacterial flora of natural mineral waters bottled in glass or in PVC containers and in view of the results obtained during their own investigations, the AA. consider it impossible to establish a limit to the charge of bacterial flora in agar at 20 degrees C and believe that any possible proposal concerning rules limiting the bacterial charge at 37 degrees C requires more detailed study and preliminary controls. In any case, they propose to apply the strictest vigilance in excluding the presence of any sign whatever of fecal contamination or of potentially pathogenic bacterial types (f.i. P. aeruginosa) in all water samples; they also suggest to require that the performance of comparative tests be extended to the stages preceding bottling. Current experimental investigations reveal that the multiplying process seems to be accelerated and more protracted in time in waters bottled in PVC containers than in those kept in glass containers.

[Assessment of the effects of alfuzosin on miction with dynamic echography]. / Valutazione degli effetti dell'alfuzosina sulla minzione mediante ecografia dinamica.

Alfuzosina has a specific antagonist effect on the alfa-1-adrenergic post-junctional receptor posed at urethral laeve musculature, prostatic capsule and vesical trgonum. It's and ideal drug the symptoms of the difficult urinary flow typical of the benign prostatic hypertrophia (IPB) and of some patologies of vesical collum to attenuate. With this work we want to demonstrate the anatomic alterations of vesical collum and prostatic urethra by means of dynamic recording mictional phase with transrectal scan carried out before and after treatment. In this study, 30 patients were divided into three groups of 10 in relation to the pathology: overtone of vesical collum, little benign prostatic ipertrophia and great benign prostatic ipertrophia have included. The patients of each group have also been divided in undergroups of five (selected with the method of double-blind) of which the first treated with alfuzosina 7.5 mg/die for 30 days and the second with placebo. Considering the limits of this study, the analysis of the results have permitted to conclude that the drug has determined a subjective improvement in all patients except those with great IPB, while the objective valuation, reached comparing scan dynamic pictures before and after treatment, has made it possible to visualize an improvement on the opening of the vesical collum and the prostatic urethra after therapy with great variations of the size of che mictional funnel and same prostatic urethra. The alfuzosina is efficacious, in the cases under examination, except the third group were the compression on the urethra was of an elevated degree.

[Spontaneous rupture of the excretory tract following renal colic in kidney malformation]. / Rottura spontanea della via escretrice a seguito di colica renale in malformazione renale.

This paper reports a group's experience in treating a case of excretory tract rupture caused by a renal colic. After careful analysis and description of this case as well as related literature analysis, they have hypothesised the dynamic physiopathological events involved in this case. In addition, they have offered clinical considerations in preventing and detecting the development of such a lesion.

[A seroepidemiological study on the level of immunological coverage in a nomadic population in Rome]. / Studio sieroepidemiologico sul livello di copertura immunitaria in una comunita nomade a Roma.

Immunization status to the three types of poliovirus, to tetanus, diphtheria and measles was evaluated in a Gypsy population living at a Roman camp. Information about demographic data and history of immunization was collected from 149 subjects and a blood sample was obtained from 86 individuals to determine antibody titres to the above mentioned infectious agents. Among the responders, only 20.8% had received at least one dose of oral polio vaccine (OPV), tetanus and diphtheria vaccine, while none was vaccinated against measles. In spite of a low immunization coverage, serological data showed high prevalence of antibodies to the three types of poliovirus (81.4% to polio type 1:94.2% to polio type 2:62.8% to polio type 3) and to measles (76.7%), while antibodies to tetanus and to diphtheria were detected respectively only in 3.5% and 0% of the individuals tested. High levels of antibodies to polio were found also among unvaccinated subjects. For these, a statistically significant positive correlation between age and number of "contact doses" from vaccinated family members was observed (r = 0.70; CI 95%: 0.27-0.90). In conclusion, this study uncovers very low levels of immunization to poliovirus, tetanus, diphtheria and measles in the study Gypsy population, and shows the effects of the secondary spread of the OPV, which probably contributed to reduce the risk of contracting the disease in unvaccinated individuals.

[Bilateral carcinoma of the breast: treatment and prevention, Review of the literature]. / Il carcinoma bilaterale della mammella: trattamento e profilassi. Revisione della letteratura.

Authors analyze 256 patients admitted in III Clinica Chirurgica of University of Rome "La Sapienza" for breast cancer, from 1982 to 1992; 16 of them had later developed another neoplasm in the opposite breast. After a retrospective study of the clinical history of all patients they came to the conclusion that bilateral neoplasm frequency is similar to that reported in world literature (1-18%) (23, 9, 2). Moreover, are analyzed the primary risk factors for a bilateral carcinoma, and they consider the utility of the prophylactic surgical treatment of the opposite breast.

[Chronic therapy with ursodeoxycholic acid in a child with Alagille syndrome]. / Terapia cronica con acido ursodesossicolico in un bambino affetto da Sindrome di Alagille.

The effectiveness of oral bile acid therapy with ursodeoxycholic acid (10 mg/kg/day) was investigated in a 10 year old boy affected by Alagille's Syndrome, a chronic cholestatic disorder due to congenital hypoplasia of the intrahepatic biliary ducts. Cholestatic and hepatonecrotic indices were measured before and during ursodeoxycholic acid therapy and 1 month after stopping and 36 months after restarting the treatment. Ursodeoxycholic acid led to a marked improvement in the cholestatic and hepatonecrotic parameters which was maintained during all the treatment phase. Pruritus and steatorrhea disappeared during the treatment with ursodeoxycholic acid. Histological examination of the liver biopsy after the treatment revealed a disappearance of the biliary plugs but without increasing he intrahepatic bile ducts. The results suggest that ursodeoxycholic acid may improve the condition of the children affected by Alagille's Syndrome, specially when the liver transplantation is required, and indicate a need for long term studies in a larger number of patients.

[Chondrosarcoma of the mediastinum: a clinical case. Diagnosis using needle aspiration]. / Condrosarcoma del mediastino: caso clinico. Diagnosi mediante ago-aspirazione.

The Authors report a rare case of mediastinal chondrosarcoma. Diagnosis, cytology and therapeutic problems are underlined.