p53 expression in cultured blood human monocytes infected with mycobacterial strains.

BACKGROUND: An upregulation of the cell-cycle associated proteins p53 and p21/Waf1/Cip1 induced by mycobacteria was previously reported. We aimed to evaluate the expression of such proteins in peripheral blood human monocyte cultures infected with strains of different mycobacterial pathogens. METHODS: The study relied on the immunocytochemical determination of p53, p21/Waf1/Cipl, bcl-2 and on the Tunel detection of apoptosis in monocytes populations cultured on four-welled chamber slides (10(6) cells/well) infected with Mycobacterium tuberculosis, M. bovis and M. avium for four consecutive days (mycobacterium/monocyte ratio 10:1). The results were expressed as mean values and SD of the percentages of stainings recorded in five fields per slide. RESULTS: The statistical analysis with Fischer test demostrated that at most sampling times the p53 and p21/Waf1/Cip1 expression and the apoptosis index were significantly higher in M. tuberculosis infected cultures than in controls (p<0.05). The M. bovis related picture diverged from the previous one for a lower p53 expression (p<0.05) at all sampling times. The M. avium infected culture values did not diverge significantly from the controls. CONCLUSIONS: The p53 and p21/Wafl/Cipl upregulation is compatible with both host defense strategies and pathogen strategies (safeguard of intracellular sanctuaries). The discrepancies among different cultures suggest a direct relationship between p53 activation and mycobacterial ability to enter host cells.

Intravesical bacillus calmette-guerin (BCG) as inducer of tumor-suppressing proteins p53 and p21 Waf1-Cip1 during treatment of superficial bladder cancer.

PURPOSE: Previous in vitro investigations recorded an inhibition of cell proliferation by BCG when added to different cell cultures. The induction of apoptosis by BCG is controversial. Our study aimed to evaluate the influence of BCG on the expression of tumor suppressing proteins p53 and p21Waf1-Cip1 and apoptosis of the urothelial cells in vivo. MATERIALS AND METHODS: Twenty-one cases of superficial bladder cancer, treated with TUR and subsequent intravesical BCG, were studied retrospectively. The assays evaluated the expression of p53 and p21Waf1-Cip1 by immunochemistry (IHC), and the presence of apoptosis by TUNEL assay. The estimates were performed, in each case, on the following specimens: one tumor sample and one non-neoplastic sample collected during the TUR which preceded the administration of BCG; one non-neoplastic sample collected 3 months after the diagnosis; and one non-neoplastic sample collected in the first 2 weeks after the completion of the treatment. Samples of 6 cancer recurrences detected during BCG were examined too. RESULTS: As usual for non-neoplastic urothelium, the pre-BCG samples displayed poor p53 and p21Waf1-Cip1 immunoreactivity. By contrast, the samples collected during and in the aftermath of BCG showed an overall increase of the expression of both proteins. The rare occurrence of apoptosis proved to be chronologically unrelated to the BCG treatment. DISCUSSION: The relationship between changes of the IHC features and BCG suggests that BCG, at least under some circumstances, can induce the activation of wild type p53 and p21Waf1-Cip1 in the urothelium. The mechanism of the BCG-p53 status interaction and its role in the antitumor activity of BCG remain to be clarified.

Direct detection of Mycobacterium tuberculosis complex and M. avium complex in tissue specimens from cattle through identification of specific rRNA sequences.

Mycobacterium Tuberculosis Direct test and Accuprobe M. avium Complex test, rely on recognition of specific rRNA sequences. The present investigation included sequentially a pre-evaluation study and a study on non-visible lesion tuberculin reactor cattle. As regards the first study, tests were performed on tissue samples from 14 reactors with tuberculous-like lesions and from as many lesion-free non-reactors, using culture findings as a reference. MTD and Accuprobe M. Avium Complex tests proved positive in 12 and 2 reactors respectively. Both tests proved negative in all non-reactors. Data consisted with culture findings. The aim of the second study was to verify the presence of mycobacteria in lymph node and tonsil homogenates from 32 non-visible lesion cattle. MTD and Accuprobe M. avium Complex test proved positive in 10 and 14 subjects respectively; no overlap was found. Our research demonstrates that low to minimal amounts of the main pathogenic mycobacteria can be reliably identified from cattle tissues by commercially available tests, which give simple, more rapid results than conventional techniques.

Rapid identification of Mycobacterium tuberculosis complex on urine samples by Gen-Probe amplification test.

The aim of the study was to evaluate the applicability to urine samples of the Amplified Mycobacterium tuberculosis Direct Detection Test (AMTD), which is currently used to identify this organism in respiratory specimens within a few hours. The study was performed on 95 patients, comprising 35 subjects with a high index of suspicion for active tuberculosis of the urinary tract and 60 subjects with evidence of non-mycobacterial disease. One urine specimen from each subject was examined by microscopy, culture and AMTD. AMTD was positive in 38 specimens and negative in 57. Assuming culture as the reference standard, the sensitivity, specificity, positive predictive value and negative predictive value of AMTD were 100%, 91.93%, 86.84% and 100%, respectively. Reassessing the discrepancies between AMTD and culture by review of patients' charts, the sensitivity, specificity, positive predictive value and negative predictive value of AMTD were 100%, 93.44%, 89.47% and 100%. The results of the study as well as the characteristics of AMTD encourage its use for the rapid recognition of urinary tract tuberculosis, although its findings should be interpreted cautiously when the clinical picture is not consistent with an active tuberculosis.

Rapid identification of mycobacteria from culture using acridinium-ester-labelled DNA probes.

OBJECTIVES: Our study used newly developed acridinium-ester-labelled DNA (AE-DNA) probes on 183 mycobacterial isolates, performing the tests on 12B Bactec vials and Loewenstein-Jensen (LJ) slants. METHODS: The probe results were verified using the conventional method as a reference. RESULTS: The probe for M. tuberculosis complex correctly identified 131 of 133 M. tuberculosis isolates, with two false negatives and no false positives, for a sensitivity of 98.5% and a specificity of 100%. The M. gordonae probe correctly identified 27 out of 27 M. gordonae isolates, with no false negatives and one false positive, for a sensitivity of 100% and a specificity of 90.9%. One hundred sixty-eight of the 183 isolates were screened in accordance with an algorithm, designed primarily for the rapid detection and identification of M. tuberculosis. CONCLUSION: The use of the algorithm considerably reduced detection time: M. tuberculosis strains were identified within 16.2 +/- 2.3 days, while identification by conventional method required 35.7 +/- 5.5 days. Probe testing resulted in a cost increase of 67.8%.

[Tolerability to terizidone (TZ) in the treatment of pulmonary tuberculosis in dialyzed patients]. / Tollerabilità del terizidone (TZ) nel trattamento della tubercolosi polmonare in pazienti dializzati.

Six dialysed patients affected by tuberculosis involving the respiratory apparatus were included in the study. Antimycobacterial treatment consisted of polychemiotherapeutic combinations using terizidone (TZ). The need to resort to this drug may be explained by the need to exclude normally used drugs (R/AMP, INH, SM) due to resistance, allergy or organ meiopragia. Five patients received terizidone at a dose of 600-900 mg/day for the entire course of treatment, whereas it was necessary to abandon treatment in one patient due to the onset of depressive symptoms which then rapidly regressed following the suspension of TZ therapy. The good level of tolerance demonstrated in the 5 remaining cases confirm the inclusion of this drug in the treatment of a limited number of cases of tuberculosis in dialysed patients.

[Contribution of the serum anti-mycobacterial IgG measurement to the diagnosis of tuberculosis. Personal cases]. / Contributo della determinazione delle IgG sieriche antimicobatteriche alla diagnosi di tubercolosi. Casistica personale.

Data of a transversal research on 36 suspected tuberculosis (tb) inpatients of St. Luigi Hospital are reported. Serum antimycobacterial antibodies Class IgG to A60 were measured by means of ELISA. Final diagnosis was tb in 19 subjects, while 9 had a tb history over different years. Sensitivity was 78.94% and specificity 75%. According to with other recent reports, our results are poorly discriminating. However, the test, if valued with other clinical data, can contribute to the ascertainment of tb.

[Different rates of theophylline clearance. A statistical study of a group of asthmatics]. / Studio delle diverse velocità di clearance della teofillina. Studio statistico su un gruppo di asmatici.

In standard clinical practice the pharmacokinetic profile of theophylline is monitored by measuring blood levels of the drug. However a more accurate picture of the metabolic pattern over time can be obtained by monitoring theophylline clearance. This technique was applied to 40 lung disease patients most (55%) of them slow metabolisers as described in the literature. In the light of this findings, cautious dosing is advisable when no data are available or clearance (difficult to apply outside a strictly specialist environment).

[Clinical aspects of respiratory pathology in scleroderma. Case reports]. / Aspetti clinici della patologia respiratoria nella sclerodermia. Contributo casistico.

Six cases of scleroderma with pulmonary involvement, either isolated or accompanied by other visceral signs, observed between 1985 and 1987 have been examined. Patients were undergoing treatment with cortisone drugs in 4 cases and azathioprine in 2. Chest X-ray documented a picture of nodular opacities in 3 cases, reticular in 2 and miliariform in 1. B.A.L. revealed a picture of prevalently neutrophil alveolitis in all subjects, while transbronchial biopsy showed a finding of macrophagic infiltration in 2 cases, and one of fibroblastic proliferation in 4. The anatomoclinical damage was reflected adequately by spirometric tests and above all by the DLCO, while Gallium 67 scan and measurement of the A.C.E. were not very sensitive.

The release of thromboxane B2 and 6-keto-PGF1 alpha following pulmonary embolism.

It has been shown in animals that acute obstruction of pulmonary artery branches is followed by an early but shortly lived increase in blood levels of thromboxane B2 and a subsequent longer-lasting increase in blood levels of 6-keto-PGF1 alpha. Our study was conducted on twelve patients with acute pulmonary embolism. Nine were treated with urokinase; three could not be given thrombolytic or anticoagulant drugs due to bleeding peptic ulcer (2 cases) or recent cerebral hemorrhage (1 case). HPLC and RIA tests were performed on arterial blood samples at diagnosis and after 1, 6, 12 and 24 hours. Findings were compared with those in a control group of 6 healthy subjects. There was a difference in prostanoid behaviour between the untreated and urokinase treated patients. Among the former mean TxB2 was significantly raised at clinical onset and began to decline after 6-12 hours, approaching the mean level found among the controls after 24 hours. In contrast 6-keto-PGF1 alpha was raised after 1 hour and gradually declined thereafter. In the subjects treated with urokinase TxB2 was already close to the mean control level after 1 hour; 6-keto-PGF1 alpha had increased after 1 hour but had returned near the control level after 12. The behaviour of prostanoids appears to match the clinical course.

[Lung diseases associated with silicosis. Study of 618 cases]. / Pneumopatie associate a silicosi. Indagine su 618 casi.

Retrospective research has been carried out on the clinical reports of 618 cases of patients suffering from silicosis admitted to the S. Luigi Gonzaga Hospital in 1973-1987 (1.4% of all pneumopathy admittances). A comparative evaluation of the five-year periods 1973-77, 1978-82, 1983-87 showed a steady reduction in the percentage of silicosis to total cases (from 1.62% to 1.55% and 1.16%). Among the 618 cases assessed, the following pneumopathies were associated with silicosis either alone or in combination: chronic obstructive bronchopneumopathy (89.4%), pulmonary tuberculosis (22.2%), bronchogenic carcinoma (8.4%), acute aspecific infections (26.1%), mycosis (0.6%), sarcoidosis (0.3%), other pneumopathies (1.1%). Comparison between the three five-year periods shows an increase in the frequency of the silicosis-carcinoma association and a fall in the silicosis-tuberculosis and silicosis-COLD associations.

Examination of 41 cases of laryngeal tuberculosis observed between 1975-1985.

Forty one cases of laryngeal tuberculosis, accounting for 0.8% of all cases of tuberculosis observed in 1975-1985, are evaluated. The mean age was 53 yrs, the male/female ratio 9:1. In all cases current or earlier pulmonary tuberculosis was noted. The laryngeal sites affected were, in order of frequency: true vocal cords, epiglottis, false cords and laryngeal ventricles, arytenoid and interarytenoid area, subglottic area. The patients were usually treated with isoniazid, rifampicin, ethambutol for 6-12 months. Two cases required excision of laryngeal lesions. The results of chemotherapy were good.

Tuberculosis of the larynx. Today.

Forty-one cases of laryngeal tuberculosis accounting for 0.79% of the TB cases observed in 1975-85 are examined. The M/F ratio was 9.2:1; the mean age 52 years 9 months. In all cases current or earlier tubercular lesions on the pulmonary parenchyma were also noted. The laryngeal sites affected were as follows, in order of frequency: true vocal cords, epiglottis, false cords and laryngeal ventricles, arytenoid and interarytenoid area, subglottic area. The patients were treated with R/AMP, INH, EMB or SM, EMB with satisfactory results. Two cases required exeresis of laryngeal lesions and 3 tracheotomy.

[Aluminum lung as an occupational disease. Case reports]. / Il polmone da alluminio come malattia professionale. Contributo casistico.

The cases are examined of 13 males, mean age 53, granted disability pensions by the Turin I.N.A.I.L. in 1975-86 as suffering from lung disease caused by the inhalation of dust and exhalation of aluminium. The patients had worked in bauxite smelting for Al2O3 production and the preparation of synthetic abrasives (8 cases), in milling using synthetic abrasives (1 case) in the cold grinding of aluminium for paint production (3 cases) and in the electrolytic processing of aluminium (1 case). In line with the latest reports in the literature the present series confirms the possibility that interstitial fibrosis can be caused by exposure to aluminium in industries other than bauxite processing. The relatively benign character of modern aluminium lung is also confirmed.

[Talcosis as an occupational disease. Case histories]. / La talcosi come malattia professionale. Contributo casistico.

The case are examined of 49 people (45 males, 4 females), mean age 60 exposed to the risk of talc inhalation in a talc processing and packing factory who were granted disability pensions by the Turin I.N.A.I.L. on the grounds of silicosis in 1975-86. Radiologically the cases presented small round patches in 28 subjects small irregular ones in 15 and large patches in 6. In functional terms, examination revealed ventilatory insufficiency that was primarily restrictive in 17 cases, obstructive in 11 and mixed in 21.

[Comparison of protective effects induced by the ultrasonic mist of nicardipine + fenoterol versus nicardipine + fenoterol + ipratropium bromide]. / Raffronto tra la protezione nei confronti della nebbia ultrasonica indotta dalla nicardipina + fenoterolo versus quella indotta da nicardipina + fenoterolo + ipratropium bromuro.

A comparison is presented of the protective effect of ultrasound mist induced by nicardipine + fenoterol and nicardipine + fenoterol + ipratropium bromide in a group of 11 patients with chronic bronchial asthma in a phase of relative clinical stability. The data collected show that the three-drug combination was more effective than the use of nicardipine and fenoterol.

[Blood copper levels in a group of patients with asbestosis]. / Tassi sierici di cupremia in un gruppo di soggetti affetti da asbestosi.

Colorimetry was used to assay blood copper levels in 13 asbestosis patients (mean age 55.5) selected by excluding all cases with an evident clinical anamnesis of dietary imbalances or other significant pathologies. The results show increased blood copper in 11 out of 13 patients, the mean level in this group being significantly higher than that in a control group (p less than 0.01). The phenomenon, already described in other interstitial lung diseases, may well be attributable to enhanced reabsorption of the metal due to the intensified activity of a copper enzyme lysyl oxidase that is involved in collagen synthesis.

[Histo-radiological and histo-bronchoscopic correlates in neoplastic pathology of the lung. Personal case series]. / Correlazioni isto-radiologiche e isto-broncoscopiche nella patologia neoplastica del polmone. Casistica personale.

Data are presented on a series of 100 lung cancer patients, diagnosed histologically and encountered in 1984-87. The percentage distribution of the various histotypes and their relationship with radiological and endoscopic aspects were examined. The data obtained are then compared with those reported in the literature.