[Two cases of acute eosinophilic pneumonia--analyses of cytokines in BALF before and after treatment].

Acute eosinophilic pneumonia (AEP) has been well recognized as sudden onset respiratory failure characterized by an increased eosinophilic cell count in the bronchoalveolar lavage fluid (BALF). Although several lines of research have demonstrated that the Th2 cytokine network is likely to play a pivotal role in the development of AEP, that mechanism has not been clearly understood yet. Previous reports published in Japan demonstrated that the BALF CD4/CD8 ratio in patients with AEP ranged between 1.1 and 5.8. In this report, we describe 2 cases of AEP in which the CD4/CD8 ratios were 0.3 and 7.8 when present. We measured IL-4, IL-5, IL-10 and IFN gamma in the BALF before and after treatment in these patients. IL-5 decreased drastically after the treatment phase, while the other cytokines did not change much. We concluded that the IL-5 concentration in the BALF is probably related to the development of AEP. In addition, we also speculated that the CD4/CD8 ratio in BALF from patients with AEP may be affected whether a patient has an atopic background or not.

Expression of syndecan-1 is common in human lung cancers independent of expression of epidermal growth factor receptor.

In order to determine syndecan-1 expression in lung cancer, we examined 115 lung cancer specimens and 17 lung cancer cell lines. Syndecan-1 was immunohistochemically stained with a polyclonal antibody in 115 paraffin-embedded specimens; 84 cases out of 97 non-small cell lung cancer (NSCLC) and eight cases out of 18 small cell lung cancer (SCLC) were positively stained. Simultaneously, epidermal growth-factor receptor (EGFR) was stained; 47 cases out of 97 NSCLC and one case of 18 SCLC were positively stained. No significant correlation was shown between EGFR and syndecan-1 expression (P=0.68). Syndecan-1 mRNA was detectable in 16 of 17 lung cancer cell lines and EGFR mRNA in nine of 17. Eight cell lines had syndecan-1 mRNA as well as EGFR mRNA. PR-39 (1 microM) and 80 pM transforming growth factor-beta(1) (TGF-beta(1)), did not increase expressions of syndecan-1 mRNA and EGFR in five lung cancer cell lines. We concluded that lung cancer had detectable syndecan-1; however, expression of syndecan-1 protein did not correlate with survival time of lung cancer patients.

Detection of Photofrin Fluorescence From Malignant and Premalignant Lesions in the Bronchus using a Full-color Endoscopic Fluorescence Imaging System.

STUDY OBJECTIVES: To detect invisible lung cancer and to determine field of laser radiation during PDT we developed a full-color fluorescence fiberscopic system. We tested the efficacy of this system in patients with various bronchial malignancies.System design: A fiber-optic endoscope was attached to a camera box containing a color ICCD camera which can detect from 400 to 700nm fluorescence in full-color. Light of average wavelength 405 nm was selected and radiated through the light channel of the fiberscope from a 300W Xenon lamp. PATIENTS AND METHODS: We examined nine consecutive patients with bronchial malignancy admitted in our hospital to receive PDT. Sixteen lesions in these nine patients were observed with white light and excitation light and the results were compared. Histological examinations were done by taking biopsy specimens and samples for pathological and cytological examination. After the diagnosis was confirmed, 2.0 mg/kg Photofrin was injected. Forty eight hours after the administration of Photofrin, observation of the bronchial wall was made using a full-color endoscopic fluorescence imaging system just before PDT. RESULTS: Bright red fluorescence from Photofrin was Observed in 14/14 bronchial malignancies: 3 squamous cell carcinoma, 9 squamous cell carcinoma in situ, 1 metastatic breast cancer and 1 metastatic islet cell tumor. Bright red fluorescence was also detected in 2/2 squamous dysplasia. Green autofluorescence was observed in the normal part of the bronchus. CONCLUSIONS: RESULTS of the present study suggest that the full-color endoscopic fluorescence imaging system can be used to detect malignant and premalignant lesions as red fluorescence against green autofluorescence with Photofrin administration, and this system has the potential to detect absence of autofluorescence in cancerous lesions.

Epidermal growth factor receptor expression correlates with poor prognosis in non-small cell lung cancer patients with p53 overexpression.

To determine whether cancer patients with tumor suppressor gene abnormality survive for a shorter time when their growth was stimulated by growth factors, we examined 290 non-small cell lung cancer (NSCLC) specimens for p53 and epidermal growth factor receptor (EGFR) protein expressions using immunohistochemical staining. The distribution of cases by pathological stage of tumor was 155 cases of stage I, 30 cases of stage II, 96 cases of stage III and 9 cases of stage IV. Pathological types were 142 adenocarcinomas, 127 squamous cell carcinomas, 17 large cell carcinomas and 4 other types of malignancy. Immunohistochemical staining was performed on the formalin fixed, paraffin-embedded materials with monoclonal antibodies DO-7 and clone EGFR.133. positive staining for EGFR was seen in 124 (42.8%) cases. More EGFR positive cases were found in squamous cell carcinomas than in non-squamous cell carcinomas (p=0.0121). Staining for p53 protein was observed in 147 (50.7%) specimens. Multivariate proportional hazard model analyses revealed EGFR protein expression as a risk factor in the patients with NSCLC (p=0.0240). Patients negative for both EGFR and p53 survived for a longer period of time (p=0.0427).

Inhibition of human small cell lung cancer cell growth by methylprednisolone.

Methylprednisolone (MP) inhibited cancer cell growth at concentrations between 0 and 2.0 mM. IC50 was 0.96 mM in H82, 0.44 mM in H345, 0.86 mM in H510A, 0.54 mM in N592 and 0.52 mM in H2081. The growth inhibition was not disturbed by addition of 1 microM RU38486. DNA fragmentation were observed in H510A and N592 at MP concentration of 0.2 mM. Glucocorticoid (GC) receptor mRNA expression was detectable only in H510A. We concluded that MP has potency as an anti-cancer agent at a high concentration. The effect was probably not through GC receptor binding, and growth inhibition was independent of apoptosis induction.

Increased serum and urinary levels of a parathyroid hormone-related protein COOH terminus in non-small cell lung cancer patients.

Parathyroid hormone-related protein (PTHrP) is expressed in a variety of human cancers including lung cancer. Three mature peptides with different COOH-terminal regions, PTHrP (1-139), PTHrP (1-173), and PTHrP (1-141), are translated from three different mRNAs through alternative splicing. In each, COOH-terminal fragment (C-PTHrP) is stable and measurable in the urine. In the present study, we measured concentrations of circulating and urinary C-PTHrP in 28 patients with primary lung cancer and normal serum calcium levels. We used PCR to evaluate PTHrP mRNA expression and its alternative splicing types in 16 lung cancer cell lines and 17 lung cancer tissues. The average serum C-PTHrP level was 38.95 +/- 19.41 pmol/l in 28 lung cancer patients, whereas that in 10 normal subjects was 26.53 +/- 9.43; the difference was statistically significant (P = 0.0065). Average urine C-PTHrP:urine creatinine ratio was 7.56 +/- 5.17 x 10(-1) pmol/mg creatinine in 28 lung cancer patients, whereas it was 4.91 +/- 1.77 in 10 normal subjects; the difference was statistically significant (P = 0.0287). C-PTHrP radioimmunoassays detected that 23% of non-small cell lung cancer patients had higher serum C-PTHrP levels, and 32% had higher urinary C-PTHrP:urine creatinine ratio than average + 2 SD of normal subjects. Reverse transcription-PCR detected PTHrP mRNA expression in 21 of 21 non-small cell lung cancer (NSCLC) samples and 3 of 12 small cell lung cancer samples. In the cancer cell lines and tissues that had detectable PTHrP mRNA, PTHrP (1-139) mRNA was found in 21 of 24, PTHrP (1-173) mRNA was found in 19 of 24, and PTHrP (1-141) mRNA was found in 23 of 24. Our results suggest that all PTHrP mRNA expression is common in lung cancers. We found that NSCLCs cancers had detectable PTHrP mRNA, and serum and urinary C-PTHrP levels in NSCLC patients were significantly higher than those in normal subjects. We concluded that NSCLC produced PTHrP more frequently, but there was no clear significance of C-PTHrP measurement in lung cancer patients for cancer detection using the present assay. We suggested that PTHrP probably plays a role similar to a growth factor or proliferation factor in lung cancer, especially NSCLC, at a level insufficient to cause humoral hypercalcemia of malignancy.

A case of cardiac sarcoidosis: significance of ventricular tachycardia originating from the septum.

A 65-year-old woman was admitted for assessment of recurrent tachycardia. Cross-sectional echocardiography showed that the anterobasal portion of the ventricular septum was thin and dyskinetic. An electrophysiologic study revealed ventricular tachycardia, during which marked fragmented potentials could be obtained from the anterior septal aspect of the right ventricle. The site of earliest activation was in the vicinity of the His bundle. A diagnosis of cardiac sarcoidosis was made by based on endomyocardial biopsy combined with the clinical manifestations. Ventricular tachycardia originating from the anterior septum may be an indicator of underlying cardiac sarcoidosis.

[Respiratory disorders in type-1 hereditary motor and sensory neuropathy].

Type-1 hereditary motor and sensory neuropathy (HMSN I) is a slowly progressive disease resulting in distal muscle weakness with atrophy, and in sensory disturbance. Restrictive lung disease and respiratory muscle failure, common in many advanced neuromuscular disorders, is not a predominant feature of HMSN-I. Recently, there have been several reports of respiratory dysfunction in patients with HMSN I, complicated by diaphragmatic weakness. In five patients with HMSN I (3 men and 2 women, mean age 55.4 yrs), we measured spirometric variables, maximal inspiratory pressure, and maximal expiratory pressure, in both sitting and supine positions. We also studied phrenic nerve conduction by cutaneous stimulation at the posterior border of the sternocleidomastoid muscle. Four of five patients had low maximal inspiratory pressure and abnormally long phrenic nerve latency. Two patients showed evidence of a restrictive lung disorder and daytime alveolar hypoventilation. All-night polysomnography in those two patients revealed periodic decreases in arterial blood oxygen saturation, and episodes of central apnea. We conclude that diaphragmatic dysfunction is not rare in HMSN I, and that maximal inspiratory pressure and phrenic nerve conduction may be useful in the early detection of phrenic nerve involvement.

bcl-2 and p53 protein expression in non-small cell lung cancers: correlation with survival time.

Apoptosis-related genes have received increasing attention in carcinogenesis, drug sensitivity, radiation sensitivity, and patient survival. bcl-2 and mutated p53 genes have been reported to inhibit apoptosis. To determine bcl-2 and p53 protein expression and their impacts on survival time in lung cancers, we studied 99 surgically resected, paraffin-embedded non-small cell lung cancer (NSCLC) specimens by immunohistochemical staining. The bcl-2 protein was expressed in 19.2% of NSCLCs. bcl-2-positive cases were found in 30. 4% of stages I and II carcinomas in 36.8% of squamous cell carcinomas. Patients with bcl-2 expression survived longer than those without. p53 protein was found in 44.4%; there was no significant difference in survival time between patients with and without p53 expression. Patients who were both bcl-2 positive and p53 negative survived significantly longer than those who were bcl-2 negative or p53 positive. These results suggest that bcl-2 protein expression can be histologically specific and stage dependent, and that the bcl-2 protein expression is potentially valuable for prognosis in NSCLC, particularly in the early stages, when bcl-2 protein expression is considered with mutant p53 protein expression.

The effects of 4-aminopyridine on focal nerve conduction block.

In order to test whether 4-aminopyridine (4-AP), a potassium channel blocker, may be of therapeutic value in demyelinating neuropathies, a focal tibial nerve conduction block with demyelination was produced in adult rats by an intraneural microinjection of potassium tellurite. Onset and recovery of the lesion were monitored by evoked compound muscle action potentials (CMAP) activated from the proximal and distal nerve one day before and 1, 4, 7, 14, 21 and 28 days after the injection. Intraperitoneal 4-AP (2 mg/kg) or buffered saline were injected prior to the potassium tellurite and 6 days per week for 28 days. The data show that 4-AP is tolerated, it does not prevent conduction block, and only has a modest effect on increasing its recovery from day 4 to 7 (91 % increase in CMAP ratio compared with control of 35%). Recovery is similar by day 28 in 4-AP treated or untreated animals. These results suggest that 4-AP will have limited use in the therapy of subacute demyelinating neuropathies.

Nerve conduction studies in the Twitcher mouse (murine globoid cell leukodystrophy).

Progression of the neuropathy in the Twitcher mouse (twi-C57BL/6J), an animal model of globoid cell leukodystrophy, was assessed with serial motor nerve conduction studies from just after birth until near death (day 45) and after hematopoietic cell transplantation (HCT). Under ether anesthesia, the tibial nerve was stimulated percutaneously at the sacral notch and at the ankle, and recordings were made from plantar foot muscles. Motor conduction velocity (MCV), distal latency, amplitude, duration and number of phases of compound muscle action potentials on proximal (pCMAP) and distal (dCMAP) stimulation were measured. In 15-19 day-old Twitcher, despite the absence of motor signs, MCV was significantly reduced, 12.8 +/- 2.8 (10) m/s (M +/- SD, No. of recordings), compared with unaffected siblings, 18.1 +/- 2.6 (21) m/s (P less than 0.01). The ratio of pCMAP to dCMAP amplitudes was reduced in the Twitcher, 0.39 +/- 0.13 (10), compared with controls 0.72 +/- 0.17 (21) and the ratio of pCMAP to dCMAP phases was increased (2.8 +/- 0.8 (10) vs 1.0 +/- 0.2 (21), P less than 0.01 for all). As neurologic signs progressed by 35-39 days, MCV became slower, 5.8 +/- 1.0 (11) m/s, pCMAP and dCMAP became smaller, but the ratio of pCMAP to dCMAP amplitudes in the Twitcher (0.55 +/- 0.36, 11) was similar to controls (0.71 +/- 1.0, 20) as was the ratio of pCMAP to dCMAP phases (1.0 +/- 0.4 vs 1.0 +/- 0.1). These results suggest that there is diffuse non-uniform slowing of nerve conduction with block especially in proximal nerve fibers initially. With HCT, mean MCV remained slow (6.7 +/- 1.2 (18) m/s, vs 34.5 +/- 3.9 (12) m/s) but motor function persisted.

The pharmacological effect of thyrotropin-releasing hormone on ataxic mutant mice.

The Rolling mouse Nagoya (RMN), Staggerer, Weaver and Reeler, all of which show hereditary ataxia, were intraperitoneally injected with 25 mg/kg of thyrotropin-releasing hormone (TRH-T) or physiological saline, and changes in the motions of these animals were observed by an Animex II and an open field method. All four strains of mice with ataxia showed improvement of ataxia and an increase in the motion volume, but these changes were not necessarily consistent in degree. Improvement of ataxia was most marked in the RMN and the Staggerer, moderate in the Weaver and slight in the Reeler, which showed enhanced tremor. The relationship between the competence of transmitting information in the cerebellum and improvement of ataxia by the injection of TRH-T aroused our interest.

Effects of 2,4-dichlorophenoxyacetic acid (2,4-D) on the contractile properties of reinnervated rat skeletal muscle.

2,4-Dichlorophenoxyacetic acid (2,4-D), a common herbicide, was administered to rats (100 mg/kg, i.p.) during 24 days to determine whether or not it is toxic to regenerating peripheral nerve and reinnervating muscle. The peroneal nerve was crushed 1 cm proximal to the extensor digitorum longus muscle and recordings made in vivo after 1 to 24 days. Functional reinnervation was observed by day 10 and recovery was similar in rats receiving 2,4-D or vehicle. Distal motor latencies and muscle action potentials returned toward normal during the 24 days in a similar manner in 2,4-D and controls. Isometric twitch tensions per muscle weight on indirect stimulation returned to intact values by 17 days, but in the 2,4-D animals they became larger (P less than 0.01) than controls by day 24. The twitch:tetanus ratios were increased at day 10 and returned toward normal values in the controls but remained increased (P less than 0.01) in the 2,4-D animals at 24 days. Similar results were obtained on direct muscle stimulation. The data suggest that 2,4-D is not toxic to nerve during regeneration or muscle reinnervation in the rat, but that it does affect both twitch and tetanus tensions suggesting proliferation as well as disruption of myofibrils.

2,4-Dichlorophenoxyacetic acid (2,4-D) does not cause polyneuropathy in the rat.

2,4-Dichlorophenoxyacetic acid (2,4-D), a component of Agent Orange, was injected intraperitoneally into adult male Fisher rats for 3-12 weeks. During the period of study gait and toe-spreading reflexes remained normal and distal motor latencies, motor and mixed nerve conduction velocities and amplitudes remained similar (P greater than 0.05) in animals receiving 2,4-D or vehicle. This study suggests that 2,4-D is not toxic to peripheral nerves in the rat.

2,4-Dichlorophenoxyacetic acid (2,4-D) reduces acetylcholinesterase activity in rat muscle.

A single dose (200 mg/kg body weight, i.p.) of 2,4-dichlorophenoxyacetic acid (2,4-D), commonly used as a herbicide, caused significant decreases in acetylcholinesterase (AChE) activity in diaphragm and other muscles of the rat. The 4S, 10S, and 16S forms of AChE were affected. The effect was maximal 15 to 24 h after injection. Choline acetyltransferase (CAT) activity was not affected. Neither AChE nor CAT activities changed in sciatic nerve from 2,4-D-treated animals. Spontaneous locomotor activity decreased dramatically 4 h after 2,4-D treatment. Myotonia that was present 1.5 h after 2,4-D injection became maximal at 2 to 6 h. Twenty-four hours after drug injection, when animals were recovering from myotonia, spontaneous locomotor activity was still depressed to 50% of control values. Prolonged distal motor latencies were observed 15 to 24 h after drug administration. AChE activity and spontaneous locomotor activity returned to control values at 48 h. Thus, 2,4-D causes a decrement of end-plate AChE, as well as behavioral and electrophysiologic changes. Decreased activity of AChE may be an early step in development of the myopathy that occurs after large dose 2,4-D.

[Studies on the behavioral pharmacology of TRH-T in cytosine arabinoside-induced ataxic mice: a comparison with genetically ataxic mice].

The effects of thyrotropin releasing hormone tartrate (TRH-T) on the behavior of cytosine arabinoside (Ara-C) induced ataxic mice were studied. The ataxic mice were prepared by injecting 50 mg/kg/day subcutaneously on the 2nd, 3rd and 4th postnatal days. Spontaneous motor activities were measured with a movement spectrum analyzer (ANIMEX-2) and the degree of ataxic gait determined by an open-field study. Cerebella from these animals were subjected to histopathologic examination at 4, 8 and 12 weeks of age. Ataxic gait became improved moderately and spontaneous motor activities increased slightly following administration of TRH-T. The effects of TRH-T on ataxia and spontaneous motility in the Ara-C injected mice varied with the age, and resembled those seen in weaver mice but not those in the Rolling mouse Nagoya. The results also suggest that the effects of TRH-T on the behavior of ataxic mice correlate with the extent of pathologic changes evoked in their cerebella.