Localization and clinical significance of thyrotropin receptor mRNA expression in orbital fat and eye muscle tissues from patients with thyroid-associated ophthalmopathy.

We have studied the cellular localization of thyrotropin receptor (TSH-R) mRNA in orbital fat and extraocular muscle tissues from patients with thyroid-associated ophthalmopathy (TAO) using Northern blot, reverse transcriptase polymerase chain reaction (RT-PCR), and in situ hybridization, and we correlated the findings with clinical estimates of ophthalmopathy. Although we failed to detect TSH-R mRNA in orbital tissues by Northern blot, TSH-R cDNA was amplified in orbital fat tissue from 13 of 25 patients with TAO and from 2 of 4 control subjects, in eye muscle tissue from 2 out of 7 patients with TAO, and in cultured orbital fibroblasts and subcutaneous fibroblasts from TAO patients. In situ hybridization showed that TSH-R mRNA was detected in cultured orbital fibroblasts as well as skin fibroblasts obtained from the patient. Furthermore, the expression of TSH-R mRNA in orbital fat tissue from patients with TAO significantly correlated with the orbital fat volume and the severity of ophthalmopathy, especially the extent of eye muscle dysfunction. These results suggest that the expression of TSH-R in the orbit, especially fibroblasts, may play a role in the pathogenesis and clinical manifestations of the ophthalmopathy in patients with TAO, although a secondary effect, involving fibroblasts in TAO is also possible.

[A case of sarcoidosis presenting with hoarseness and dysphagia due to glossopharyngeal and vagus nerve paresis].

The patient was a 20-year-old female who complained of hoarseness and dysphagia. Chest X-ray showed bilateral hilar lymphadenopathy. Sarcoidosis was diagnosed histologically on the basis of granuloma without necrosis, by transbronchial lung biopsy (TBLB). Bronchofiberscopic findings revealed no granuloma of the vocal cords. Examination of the central nervous system with MRI identified no abnormalities. Hoarseness and dysphagia were thought to have been caused by glossopharyngeal and vagus nerve paresis. These signs improved markedly after two weeks of steroid therapy. This is a rare case of sarcoidosis associated with glossopharyngeal & vagus nerve paresis.

[Mechanism of efficacy of erythromycin on diffuse panbronchiolitis--effect of erythromycin on cytokine mRNA expression in human whole blood model].

Recently, "low-dose and long-term" erythromycin (EM) has been reported to be effective in treatment of diffuse panbronchiolitis (DPB), but its mechanism is still obscure. We studied the effect of EM on cytokine mRNA expression by using LPS-stimulated human whole blood as an experimental vivo model. IL-8 mRNA was expressed in biphasic fashion with peak expression at 6 hours and 20 hours from the start of LPS stimulation. When whole blood was pretreated with EM (2 micrograms/ml) for 1 hours. IL-8 mRNA expression was depressed at 20 hours (p < 0.025) from the start of LPS (1 microgram/ml) stimulation. However, when pretreated for 12 hours, it was not depressed. EM (2 micrograms/ml) also depressed IL-1 beta (p < 0.025) and TNF alpha (p < 0.05) mRNA expressions at 6 hours from the start of LPS stimulation. From the above results, it was suggested that the direct inhibition of IL-1 beta and TNF alpha production by EM resulted in subsequent depression of production of IL-8 that is a potent chemotactic factor for neutrophil, and consequently, EM acts to protect the bronchiole tissues of DPB patients from destruction by proteolytic enzymes released from neutrophils. This assumption seems to be supported by our previous observation that when patients with DPB were treated with EM a marked decrease in number of neutrophil in bronchoalveolar lavage fluid (BALF) was accompanied by clinical and radiographic improvement.

Rapid and sensitive diagnosis of cytomegalovirus and Pneumocystis carinii pneumonia in patients with haematological neoplasia by using capillary polymerase chain reaction.

We attempted the simultaneous detection of cytomegalovirus DNA (CMV-DNA) and Pneumocystis carinii (carinii-DNA) in sputum samples obtained from 20 patients with haematological neoplasm with pneumonia, using rapid cycle DNA amplification (capillary PCR). We used a thermal cycler for capillary PCR which featured recirculation of hot air for rapid temperature control of 10 microliters reaction samples in thin glass capillary tubes. We extracted DNA from patients' sputa using a simple method. A comparison of the results obtained using the phenol extraction-ethanol precipitation method and those obtained using our simple method was made, and demonstrated complete agreement between the two. For detection of CMV-DNA and P. carinii-DNA with capillary PCR it was not necessary to vary temperature setting based on the primers used. Therefore, capillary PCR was used for the simultaneous detection of CMV and P. carinii. After amplification, the total time required for which was 20 min, amplified products were electrophoresed on agarose gels and visualized with ethidium bromide. Product sensitivity was higher with capillary PCR than with conventional PCR. We conclude that capillary PCR amplification is a valuable tool for rapid and simple diagnosis of CMV and P. carinii pneumonias.

Rapid diagnosis of cytomegalovirus and Pneumocystis carinii pneumonia by using the capillary polymerase chain reaction.

We attempted to detect cytomegalovirus DNA (CMV-DNA) and Pneumocystis carinii DNA (P. carinii-DNA) in sputum samples of 18 hematological neoplasm patients with pneumonia, using rapid cycle DNA amplification. A thermal cycler based on recirculating hot air was used for rapid temperature control of 10-microliters samples in this glass capillary tubes. After a total amplification time of 15 min, the amplified products were electrophoresed on agarose gels and visualized with ethidium bromide. In three cases, CMV-DNA was detected at about the time the pneumonia occurred. These patients were successfully treated with ganciclovir in the early stages of infection and CMV was not detected by the virus culture method. In four other cases, P. carinii-DNA was detected in their sputum samples but not detected by Grocott staining. These four cases of P. carinii were successfully treated with sulfamethoxazole-trimethoprim. For detection of CMV-DNA and P. carinii-DNA using the capillary polymerase chain reaction (PCR), a different temperature setting base on the primer difference was not necessary. Therefore, capillary PCR was performed at the same time for detection of CMV and P. carinii. We conclude that capillary PCR amplification is a valuable tool for rapid diagnosis and early treatment of pneumonia due to CMV and P. carinii.

[A case of encephalopathy accompanying Mycoplasma pneumoniae pneumonia in an adult].

We experienced a 39-year-old male who developed neurological complication during a course of Mycoplasma pneumoniae pneumonia. The diagnosis of M. pneumoniae pneumonia was made on the basis of elevation of specific antibody (CF) titer in convalescent serum. Electroencephalogram showed diffuse damage in the brain, but no other abnormalities were not found on brain CT-scan and MRI. In the cerebrospinal fluid, the number of cells did not increase and the M. pneumoniae CF titer was not elevated. From these results, we concluded that encephalopathy in this patient was raised by an allergic reaction of the brain tissue to M. pneumoniae antigen. Until now, encephalitis or meningoencephalitis accompanied with M. pneumoniae infection has been reported by many investigators, but reports on encephalopathy due to M. pneumoniae are few. Therefore, we reported herein a case of encephalopathy following Mycoplasma pneumoniae pneumonia with several references.

[Detection of Chlamydia pneumoniae by polymerase chain reaction].

We aimed at developing a method for early detection of Chlamydia pneumoniae (C. pneumoniae) from clinical specimens. For this purpose, we amplified C. pneumoniae-specific DNA fragments by polymerase chain reaction. A pair of 24 mer of oligonucleotides which were complementary to the sequences within the region encoding the major outer membrane were synthesized and used as primers. As the results, all three standard strains of C. pneumoniae (AR-39, TW-183 and AR-388) were identified by the detection of the amplified products of 174 base pairs, while each strain of Chlamydia trachomatis and Chlamydia psittaci and a total of 11 strains of bacteria and a strain of Mycoplasma pneumoniae were not. Thus, the present method was found to provide a very high specificity and also a high sensitivity of a detectable level of 10 x 10(-9) inclusion-forming units.

[Rapid diagnosis of cytomegalovirus pneumonia and Pneumocystis carinii pneumonia by using polymerase chain reaction DNA amplification].

We attempted to detect cytomegalovirus DNA (CMV-DNA) and Pneumocystis carinii DNA (carinii-DNA) in urine, blood and sputum samples of 16 leukemia patients with pneumonia, using the polymerase chain reaction (PCR). Synthetic oligonucleotide primer pair were used to amplify DNA from the major immediately genes of CMV and genes for the large subunit of mitochondrial ribosomal RNA of P. carinii. Amplified products were detected by gel electrophoresis. In two cases, CMV-DNA was detected at about the time the pneumonia occurred, and in one of the two cases, CMV-DNA was detected in the sputum sample. This patient was treated immediately with ganciclovir. After ganciclovir treatment, clinical and biochemical signs of CMV pneumonia disappeared. In three cases, carinii-DNA was detected in their sputum samples. In their blood and urine samples, carinii-DNA were not detected. This three cases were treated with sulfamethoxazole-trimethoprim and successfully treated episodes of P. carinii pneumonia. We conclude that PCR amplification may be a valuable tool for rapid diagnosing CMV pneumonia and P. carinii pneumonia.

[A case of pneumonia due to Mycoplasma pneumoniae accompanying high adenosine deaminase activity in pleural effusion].

We experienced a 5-year-old male case of Mycoplasma pneumoniae pneumonia accompanying Adenosine Deaminase (ADA) activity in pleural effusion. Chest roentgenograms revealed the infiltration in the left upper lung field and the left pleural effusion. In serum, the M. pneumoniae CF titer increased to 1:512. The pleural effusion was yellowish in color, with a specific gravity of 1.030, protein 3.7 g/dl, glucose 101 g/dl, and ADA 50 IU/l. Pleural effusion accompanying M. pneumoniae pneumonia is rare, and the high ADA activity in this case has been reported only in one other case. This is a report of a high activity of ADA in the pleural fluid by M. pneumoniae pneumonia.