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Probiotic microorganisms to be used as biotherapeutic agents have to resist the rigors of the upper human gastrointestinal tract. In this study we evaluated the acid tolerance in vitro and the fecal recovery in vivo after oral administration of a Saccharomyces cerevisiae strain to healthy volunteers. At the lowest pH value (pH 1.0) the yeast load in tablets decreased slightly. From pH 1.0 to pH 7.0 the release of S. cerevisiae in buffer solutions increased. The selected yeast strain showed good tolerance to low pH which mimic the gastric environment. After one month of treatment at a dose of 100 million cells per day, S. cerevisiae grew from the feces of 6 (37.5%) of the 16 healthy, treated volunteers. Based on the results of the present experiments the yeast studied can be considered a strain that tolerates adverse conditions comparable to those of the human gastrointestinal tract, and when administered orally may colonize the bowel of healthy volunteers and even replace resident Candida species.

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In August-September 1998 an Italian medical team, composed of a plastic surgeon, an anaesthesiologist, an infectious diseases specialist and a psychologist, assisted by a local non-government organization (Narripokkho), studied a cohort of Bangladeshi women injured by sulphuric acid. The goals of the mission were: (1) recognition of the magnitude of the phenomenon of acid being intentionally thrown at women; (2) evaluation of the type and severity of burns; (3) preparation of a tentative schedule of surgical treatment of the lesions; (4) organization of local facilities; (5) training of doctors and nurses.

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The introduction of HAART has changed the nutritional status of HIV patients. In the pre-protease inhibitor (PI) era, more than 60% of HIV-positive persons presented with protein energy malnutrition (PEM) and vitamin and mineral deficit. This caused progressive physical-metabolic wasting (wasting syndrome/cachexia) and increased susceptibility to opportunistic infections and drug toxicity. PEM was a concurrent cause in 80% of deaths attributed to AIDS. Since 1996, the year in which PIs were introduced, the number of patients dying as a result of AIDS has decreased by two thirds, and cachexia is no longer the AIDS terminal phase in developed countries. But different patterns of nutritional status changes have appeared in association with the use of newer anti-HIV therapies and with longer survival of HIV-infected patients. A new clinical and laboratory syndrome--lipodystrophy syndrome--now affects patients receiving PI-based therapy. This syndrome consists of changes in body shape that are caused by an abnormal redistribution of fat. Fat accumulates in the abdominal area (truncal and visceral obesity), in the axillary pads (bilateral symmetric lipomatosis), and in the dorsocervical pads ("buffalo hump," "bull neck") but decreases in the legs, arms, and nasolabial and cheek pads (peripheral lipodystrophy). Hyperlipidemia and insulin resistance are also frequently present (metabolic syndrome X). Pathogenic mechanisms of lipid and fat tissue disturbances are discussed in this article, and the clinical approach to patient management and therapeutic options for lipodystrophy and lipid dysmetabolism is evaluated.

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The circadian rhythms of plasma growth hormone (GH), insulin-like growth factor type I (IGF-I), cortisol, adrenocorticotropic hormone (ACTH), thyroid-stimulating hormone (TSH), and prolactin (PRL) were evaluated in 13 HIV-seropositive patients (8 males and 5 females; mean age [+/-SD], 30 +/- 5 years), classified as CDC C2. Sixteen clinically healthy subjects (9 males and 7 females; mean age [+/-SD], 32 +/- 8 years) were chosen as control group. Samples were taken every 4 hr from 04:00 to 20:00 and every 2 hr from 20:00 to 04:00. Plasma GH was evaluated by IRMA procedure, plasma IGF-I by RIA (after separation of soluble IGF-I from IGF-I-binding proteins, using acid-ethanol extraction), plasma cortisol by a solid-phase RIA, plasma ACTH by double-antibody RIA, and serum TSH and serum PRL by a solid-phase two-site fluoroimmunometric assay. Rhythmometric data were analyzed by single and population mean cosinor analysis; the comparison of the parameters of the rhythm between patients and controls was carried out by the mesor test and the amplitude-acrophase Hotelling test. Alterations of the circadian pattern of GH, IGF-I, cortisol, ACTH, TSH, and PRL were demonstrated in HIV-seropositive patients. In fact, the circadian profiles of these hormones were clearly flattened and no statistically significant 24-hr rhythm was detectable (with the exception of cortisol). These results are consistent with the hypothesis that alterations of the circadian temporal structure may already be present in HIV-seropositive patients without wasting and infectious complications.

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Rhodococcus equi is a facultative intracellular, obligate aerobe, partially acid fast, gram-positive pathogen that causes cavitary pneumonia in animals and immunocompromised humans. We describe 8 cases of R. equi pneumonia in patients with advanced HIV infection (CD4 counts less than 100/mm3), 7 males and 1 female (mean age 30.8 years), observed between 1991 and 1994. A history of exposure to farm animals was found in 4 patients. The most common presenting symptoms were fever, malaise, dyspnea, cough and hemoptysis, chest pain and weight loss. Chest x-rays showed tipical focal area of consolidation throughout the lung (3 upper, 3 lower and 2 middle fields) associated with cavitation in 4 cases. The definitive diagnosis in our hands was delayed only in the first case in which conflicting data resulted from blood culture (Bacillus sp. isolation) and sputum examen (acid-fast bacterium in the Ziehl-Neelsen stain). Final microbiological diagnosis depended on blood cultures (n=5), bronchoalveolar lavage (n=1), sputum (n=1), lung biopsy (n=1). All the patients were treated with prolonged courses of antibiotic therapy (259 days, range 120-340 in 6 dead patients; more than one year and two months respectively in two patients alive). According to microbial susceptibility TMP/SMX, vancomycin, imipenem, rifampin, aminoglycosides, macrolides and quinolons were more frequently used. Resistant R. equi mutants were selected during therapy with TMP/SMX (n=2), rifampin (n=1) and erythromycin (n=1). Five patient underwent pulmonary lobectomy after exclusion of metastatic bacterial lesions. Only 2 patients are alive, one after 365 days of antibiotic therapy and upper lung lobectomy, one after 60 days of antibiotic therapy. Optimal antimicrobial therapy and the role of surgery remain, in our experience, uncertain.

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The beneficial effect of disaccharides, lactulose and lactitol, in prevention and treatment of hepatic encephalopathy is well established but their use in combination with neomycin is still controversial. We studied in vitro the fecal bacterial growth, acid and gas formation in presence of lactitol (beta-galactoside-sorbitol) and neomycin alone or in combination. The results indicate that neomycin only inhibits the growth of susceptible bacteria (E. coli, Staph. aureus) which, conversely, are poor lactitol fermenters. The resistant organisms (Lactobacillus acidophilus, Clostridium perfringens) that are efficient disaccharide fermenters continue to metabolize lactitol still when antibiotic is added. Addition of lactitol 10% increased the inhibitory effect of neomycin on bacterial growth by 25-50% within 60-70 min. These preliminary data suggest that lactitol and neomycin may have additional or synergistic effects in vivo when used together in presence of favourable intestinal microbial environment.

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During the last eighteen years (1970-1987) at the Infectious Diseases Clinic of the University of Pavia, Ospedale Policlinico S. Matteo, IRCCS, Pavia (referral Center for hepatitis in our district: 502534 inhabitants) we observed 4238 patients (2706 M = 63.8%; 1532 F = 36.2%) admitted with presumptive diagnosis of hepatitis. The male to female sex ratio was 1.78 and average age was 38 (1-90) years. Acute viral hepatitis was diagnosed in 3238 patients (76.4%), 1960 of which were males (60.5%) and 1278 (39.5%) females, with an average age of 35 (1-88) years. The possible route of transmission was: drug addition in 487 patients (15%), blood transfusion in 464 (14.3%), other (sexual, professional, familiar) in 332 (10.3%), unknown in 1955 (60.4%). Chronic hepatitis (CH) was diagnosed according to the European Association for the Study of the Liver (EASL) and to the International Association for the Study of the Liver (IASL) in 848 patients (20%), 704 M(83%) and 144 F (17%) with an average age of 48 (2-90) years. 463 patients (54.5%) were biopsied during admission, 385 (45.5%) received definitive diagnosis by clinical and previous histologic records. CAH was found in 268 (57.9%), CPH in 161 (34.8%) and CLH in 20 (4.3%) patients. Other liver diseases (steatosis, cirrhosis, HCC) were identified in 152 subjects (3%). The prevalence of A, B, NANB and Delta hepatitis virus and HI virus in the acute disease was respectively of 5.4%, 54.8%, 33.9%, 0.28% and 0.77%. In CH the HBV aetiology accounted for 49.1%, NANB virus for 44.5%, co/super infection with HDV for 15%. Among factors involved in pathogenesis of chronic hepatitis we focused attention on drug addition which was found in 129 (28.7%) patients, blood transfusion in 70 (15.6%), HIV infection in 35 of 166 (21.1%). The data still demonstrate the high prevalence of HBV aetiology of CH and existence of co-factors in the pathogenesis of chronicity. The lack of markers for NANB infection persists as the main problem in the diagnosis of liver disease. This work was supported by grant 40% from M.P.I.: "Epatiti virali acute e croniche"....

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We evaluated the susceptibility of 618 strains of yeasts belonging to the genus Candida, isolated from various biological specimens. Amphotericin B, Nystatin, Fluorocytosine, Clotrimazole, Miconazole, Econazole and Ketoconazole were tested with the Kirby-Bauer method, modified according to Drouhet and Dupont. The polyenes Amphotericin B and Nystatin and the antimetabolyte Fluorocytosine were the most effective drugs, while the imidazole derivatives (Clotrimazole, Miconazole, Econazole and Ketoconazole) generally showed a lower activity. We emphasize the importance of routine evaluation of the susceptibility of isolated yeasts to the main antimycotic drugs.

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We measured the chemiluminescence (CL) of human leukocytes (PMNs) exposed to different concentrations of Clindamycin (1; 5; 1 mcg/ml) using a standardized luminolamplified reagent-instrument methodology (Auto Picolite 6500 Luminometer and ZAP/Picolite Kits, Packard Instruments Co. Drowners Grove, IL. USA). Cells obtained from healthy donors were also tested for chemotaxis with Boyden chambers, for phagocytosis and for killing activity against Staphylococcus aureus by the agar-medium culture plate technique. Clindamycin does not induce significant variations of the CL response in whole blood, or changes in phagocytosis and killing activity. On the contrary, concentrations of drug corresponding to therapeutically obtainable levels significantly increase light emission by isolated cells. A concentration effect was seen on leukotaxis, that was increased, but not significantly, at 1 and 5 mcg/ml and decreased (P less than 0.01) at 10 mcg/ml. CL assay is a simple, sensitive, reproducible technique to assess the PMNs functions during antibiotic therapy.

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This review covers 42 patients with fulminant hepatitis treated at the Clinic of Infectious Diseases of University of Pavia over a period of 11 years (1975-1985). This group is 1.9% of the 2181 subjects hospitalized for viral hepatitis, and consists of 25 males (59.5%) and 17 females (40.5%) with a mean age of 44 years. 26 patients (62%) had a virus B hepatitis, whereas in the remaining 16 the non-B type was diagnosed. B hepatitis had an overall prevalence of 56.4%. No case of fulminant hepatitis showed prothrombin levels above 23 (overall average 14%) and bilirubin below 17 mg/dl (overall average 24.5 mg/dl). SGOT and SGPT values were 1684 and 1868 UI/l respectively. 24 patients (57.1%) developed coma after admission to hospital and 18 had this diagnosis upon entry. History evidenced a heroin habit for 6 patients (14.2%); antiblastic treatment for 13 (30.9%); a preexisting liver disease for 7 (16.6%); other major pathologies for 6 (14.2%); steroid anabolizers, barbiturates + pirazolone derivatives and oral contraceptives use for 4 (9.5%) respectively. The overall mortality rate was 90.5%. 17 subjects (40.4%) were treated in an intensive care unit with a mortality rate of 75%.

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Type and number of specific diagnostic procedures; type, time and exitus of therapy were studied in the management of 335 hospitalized patients with Salmonella infection during a nine/years period (1976-1984). Sixty-three cases of S. typhi (18.8%), 26 of S. paratyphi B (7.7%), 48 of S. typhimurium (14.3%) and 198 other Salmonella species (59.1%) were identified. Positive cultures were obtained from blood in 18.8% of cases. In typhoid fever bacteriemia was detected in 60.3% of cases. The sensitivity of microorganisms to antibiotics was detected. Of 281 (83.8%) patients treated with "major" antibiotics (CAF, Ampicillin or Co-trimoxazole) clearance of Salmonella was obtained in 171 (60.8%). Stool "acidification" with lactulose was attempted in 54 patients (16.2%), of which 20 (37%) became negative. Charges of the bacteriologic cultures, serological tests, antibiotics and supportive therapy were estimated in order to assess the cost/benefit ratio of patients care. Length of stay and chemotherapy were respectively 20.7 (11-18) and 11 (7-17) days. Maximal cost/case ratio occurs in diagnosis for stool cultures, in therapy for Ampicillin. The introduction of an economic approach to use diagnostic and therapeutic procedures has significantly improved the management of Salmonella infections from 1976 to 1984.

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Among the large spectrum of pharmacological activities of flavonoids, play an important role the recently investigated properties involving the arachidonic acid metabolism. In order to clarify the mechanisms of "cytoprotection" of the 3-palmitoyl-(+)-catechin (Palm-cat), a new flavonoid compound (C31 H44 O7) we have studied in experimental hepatitis of the rat, induced by Galactosamine (Ga1N) and E. coli 055:B 5 endotoxin (LPS), hepatic cAMP and cGMP, transaminases, bilirubin and endotoxemia. The Palm-cat significantly increases cyclic-GMP levels in the liver, whereas reduces or slightly modifies the cAMP. Transaminases and bilirubin values increase both in controls and flavonoid treated rats. The flavonoid significantly decreases the frequency of endotoxemia. These effects suggest that RES and hepatocytes functions, immune and inflammatory response can be affected in liver disease by flavonoids via cyclic nucleotides regulation.

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The Authors wish to examine the protective effects which a period of pre-treatment with thymostimulin, would have on endotoxin hepatitis, induced in thymectomized and non-thymectomized animals. The test showed that the histological picture and the degree of endotoxinemia, measured with the Lymulus test, benefited from treatment with a thymic extract. This therapy was effect (and was statistically significant) in obtaining an immunorestoring effect (in the thymectomized mouse) and in inducing an immunostimulating effect (in the normal mouse).

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Alterations of the specific enzymes located in the cell membranes might promote changes of the cyclic nucleotides ratio which is regulatory in growth stimulation of hepatocytes. Bacterial toxins, hormones and drugs affecting cyclic nucleotides system can interfere with this process in liver diseases. The Authors have determined hepatic cAMP concentrations by means of cAMP radioimmunoassay, in vivo, in rats treated with cholera toxin (CT), E. coli endotoxin (LPS) and glucagon. CT (0.15 mg i.v./rat/4th hr) and glucagon (0.8 mg/i.v./10') (significantly p less than 0.01) increase liver cAMP from 1.72 n mol/g wet wt (controls) to 2.62 and 4.13 n mol/g wet/wt respectively. Serum transaminases levels were unmodified. LPS (1.5 mg/hg/i.v./3th hr) significantly (p less than 0.01) raises serum transaminase activity while liver cAMP was not affected. Kinetics study however show that LPS, at lower dosage (0.5 mg/hg i.v.) enhances liver cAMP before cytotoxicity is observed.

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Through the use of the Limulus test research has been carried out on gram-negative endotoxin in patients with hepatic cirrhosis, chronic hepatitis, acute hepatitis, and in a control group. The positivity of this test in patients with cirrhosis and chronic hepatitis was 93.3% and in cases of acute hepatitis it was 90.9%. The effect of the combined administration of lactulose and paromomycin on endotoxin blood levels has been evaluated in a group of 9 patients with acute hepatitis, 8 with cirrhosis, 1 in hepatic coma, and 1 patient with chronic persistent hepatitis: in 18 of the 19 patients the Limulus test became negative. The results have been discussed in relation to clinical and laboratory data, and to recent data concerning the interaction between intestinal bacterial flora, endotoxin, and liver. Hypotheses have been proposed regarding the hepatocellular c-AMP mediated mechanism of endotoxin action.

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