Increased expression of tumor necrosis factor-alpha, interleukin-6, platelet-derived growth factor-B and granulocyte-macrophage colony-stimulating factor mRNA in cells of bronchoalveolar lavage fluids from patients with sarcoidosis.

Cytokines released from activated alveolar macrophages and T-lymphocytes affect the accumulation of monocyte-macrophage-lineage cells and therefore play an important role in the formation of sarcoid granuloma. Although it is likely that certain monokines and lymphokines are involved in the development of sarcoid granulomas, the evidence for this is not unequivocal. In an attempt to clear critical cytokines in the development and maintenance of sarcoid granuloma, we have measured the level of seven cytokine mRNA (TNF-alpha, IL-6, IL-8, TGF-beta, PDGF-B, IFN-gamma, and GM-CSF) in cells obtained by BAL from sarcoidosis patients and normal subjects. To detect cytokine mRNA, we employed a reverse transcription-polymerase chain reaction. We report that the levels of TNF-alpha, IL-6, PDGF-B and GM-CSF mRNA were significantly increased in BAL cells from the patients with pulmonary sarcoidosis compared to controls. No significant differences were observed in the mRNA expression of IL-8, TGF-beta and IFN-gamma. A significant correlation of the expression of the mRNA levels of seven cytokines in the same patients with sarcoidosis was observed between IL-8 and TNF-alpha, PDGF-B, and IL-6, IL-8 and IL-6 and TFN-alpha and PDGF-B and IL-8. This finding indicates that at least these four cytokines are involved in the cytokine network at the local alveolar site of chronic granulomatous inflammation. This study adds a report to the literature that supports a role for cytokine, TNF-alpha, IL-6, PDGF and GM-CSF in particular, in the promotion and maintenance of sarcoid granulomatous inflammation.

Alterations in telomeric repeat length in lung cancer are associated with loss of heterozygosity in p53 and Rb.

In the two-stage model of controlling cellular senescence in cultured human fibroblasts, retinoblastoma (Rb) and p53 proteins may be key factors regulating the mortality stage 1 mechanism. In addition, the critical loss of telomeric DNA due to the end-replication problem may result in the mortality stage 2 mechanism. Cells which acquire telomerase activity can overcome the M2 mechanism by stabilizing telomere length and thus become immortal (telomere hypothesis). At present it is known whether cellular immortality is a prerequisite for all human cancers. To investigate this question and the applicability of the two-stage model to human cancers, we analysed the relationship between alterations of telomere length and other genetic changes in lung cancer. Among 60 primary lung cancer tissues, telomere length alterations were observed in 16 tumors (26.7%) including 14 with short and two with elongated telomeres. Ten of them revealed allelic loss of both p53 and Rb genes, and remaining six showed no abnormalities in both genes. We propose that inactivation of both p53 and Rb genes may promote cell divisions causing telomere shortening in lung cancer as in the two-stage model, while there may be another pathway to overcome both M1 and M2 mechanisms, especially for adenocarcinoma.

[A case of severe bronchial asthma following first use of timolol ophthalmic solution].

A 65-year-old woman with stable atopic bronchial asthma received a prescription for timolol maleate eye drops (0.25% solution, one drop twice a day to both eyes) for glaucoma. On that evening, ten minutes after administration of the first application of timolol, the patient experienced wheeze and slight difficulty in breathing. Within the next ten minutes, her respiration became obviously asthmatic. Her symptoms progressed rapidly. When she arrived at our hospital, reduced respiratory sounds, cyanosis and disturbance of consciousness were observed. After treatment with aminophylline, hydrocortisone, epinephrine and oxygen inhalation, her symptoms completely recovered. In a provocation test, fifteen minutes after application of timolol, wheeze and dyspnea were induced following an increase in respiratory resistance. After the asthma attack, transient hypoxemia, bradycardia and decrease in blood pressure were also observed.

Rapid detection and identification of mycobacteria in sputum samples by nested polymerase chain reaction and restriction fragment length polymorphisms of dnaJ heat shock protein gene.

In the diagnosis of mycobacterial infection, more than 4-8 weeks is required to identify the species of mycobacterium responsible for an infection. Therefore, the development of a method for the rapid detection and identification of mycobacteria is necessary for selecting an optimal therapeutic plan early in the patient's course. For this purpose, we developed a method combining a nested polymerase chain reaction (nested PCR) procedure and a restriction fragment length polymorphisms (RFLP) analysis of the dnaJ gene of mycobacteria, which codes for a heat shock protein. The PCR procedure allowed the sensitive detection of mycobacterial DNA in clinical samples. Using only 10 femtograms of mycobacterial DNA as a reaction mixture, a detectable band of target DNA segments could be yielded on an agarose gel. This indicates that even with a single genome amount, the PCR is able to detect mycobacteria. The RFLP analysis of the PCR products allowed us rapidly to distinguish the strains belonging to the M.tuberculosis complex from 11 different strains of nontuberculous mycobacteria. Within 2 days, the method is able to identify the mycobacterial species present in the sputum. Moreover, it has the advantage of not requiring the use of radioisotopes, which strongly enhances its clinical usefulness.

[Clinical studies and sputum levels of ceftriaxone once daily administration in respiratory tract infection].

Ceftriaxone (CTRX), a new cephalosporin, was investigated by once daily administration for its clinical efficacy and safety on respiratory tract infections. The results obtained are summarized as follows: 1. Clinical responses to CTRX of a total of 39 cases with respiratory tract infections were excellent in 12 cases, good in 23, fair in 3, poor in 1 with an efficacy rate of 89.7%. Against acute bronchitis, lung abscess, bronchiectasis, chronic bronchitis and obstructive pneumonia, efficacy rates were 100%. 2. Serum levels and urinary excretion rates of CTRX were investigated in 2 cases after intravenous drip infusion of the drug at doses of 1 g and 2 g, respectively. Although urinary excretion rate tended to decrease with the deterioration of renal functions, prolongation of serum half-life was slight in those patients with normal liver function. In 1 case, it remained at 1.9 micrograms/ml at 12 hours and in another at 0.9 microgram/ml at 22 hours in sputum. According to the results, it appears that once daily administration of CTRX is effective and well tolerated in patients with acute respiratory infections.

Frequency of 6-thioguanine-resistant T cells is inversely related to the declining T-cell activities in aging mice.

The frequency of clonable 6-thioguanine-resistant (6-TGr) splenic T cells increased moderately with age in female BALB/c mice ranging in age from 3 to 32 months; however, the correlation between the frequency of clonable 6-TGr cells and age was weak. Those clonable 6-TGr T cells were deficient in hypoxanthine/guanine phosphoribosyltransferase (HGPRT) activity and sensitive to hypoxanthine/aminopterin/thymidine medium, as in the case of HGPRT-deficient L5178Y mouse lymphoma cells. When splenic T cells of individual aging mice were assessed simultaneously for the frequency of clonable 6-TGr T cells and for their ability to produce interleukin 2 or to proliferate in response to mitogenic stimulation, an inverse correlation was observed. These results indicate that the frequency of 6-TGr T cells is more closely related to physiologic age than chronologic age. This would mean that the frequency could be used as an index of physiologic age and that the T cells could serve as a cellular model relating gene alterations to physiologic age.

Evaluation of a new bronchodilator, Formoterol, using biochemical parameters.

The authors administered orally 40, 60 and 80 micrograms of Formoterol which was developed as a new bronchodilator and 4 mg of Salbutamol which is considered to be a beta 2-selective drug, to healthy adults. Pulse rate, blood pressure and blood levels of cyclic GMP, free fatty acid, cyclic AMP, glucose and lactic acid were measured in order to evaluate efficacy and dosage of Formoterol and to compare efficacy of this drug with that of Salbutamol. As a result, it is considered that Formoterol has a strong dose dependent beta 2-effect with 40 micrograms of Formoterol almost as high as 4 mg of Salbutamol in beta 2-effect. In addition, Formoterol has almost no alpha-effect and its beta 1-effect can be considered negligible in the doses of 60 micrograms and 80 micrograms.

Influence of age on the production and regulation of interleukin-1 in mice.

The decrease in T-cell proliferation with age is due, in part, to the decline in the production of IL-2. Since IL-1 is needed to trigger IL-2 production, we determined the IL-1 producing capacity of peritoneal macrophages of young (2-4 months) and old (24-26 months) BALB/c and C57BL/6 mice. Mice were stimulated with LPS, and their peritoneal macrophages were obtained 3 days later, purified, and assessed for IL-1 production by coculturing them with splenic T cells at a ratio of 1:5 in the presence of LPS. Supernatants were obtained 4 days later when the PGE2 and IL-2 activities were minimal and IL-1 activity maximal. IL-1 activity was assessed for their ability to augment the proliferative activity of indicator thymocytes in their response to PHA stimulation. The results revealed that (i) IL-1 production by cells of old BALB/c and C57BL/6 mice is reduced to about 40% and 30% that of young mice, respectively; (ii) indomethacin enhances IL-1 production by cells of both young and old mice to the same extent; and (iii) reduction in the IL-1 producing capacity by cells of old mice results from altered activities of both the IL-1 producing peritoneal macrophages and the augmenting T cells.

Immunologic basis for susceptibility to infection in the aged.

Age-related immune dysfunction contributes to the vulnerability of old individuals to infection, e.g., animal model studies demonstrate the association between age-related decline in T cell-dependent immunologic responses and the decline in resistance against viral, bacterial, and parasitic infections. This review briefly describes age-related changes in the immune system at the systemic, tissue and cellular levels. At the systemic level, emphasis is on polymorphic effects of aging; at the tissue level, emphasis is on the vulnerability of primary tissues engaged in the generation of antigen-responsive cells and on the difference in the onset and rate of changes between different peripheral tissues of the system; and at the cellular level, emphasis is on the qualitative changes at the surface receptor, cytoplasmic and nuclear levels.