Absence of highly homologous sequence to HTLV-I in Japanese multiple sclerosis

We tried to detect HTLV-I-related sequences in Japanese patients with multiple sclerosis with a highly sensitive method that employs the polymerase chain reaction (PCR) of genomic DNA followed by Southern blot hybridization analysis. To amplify HTLV-I sequences, we used primers for LTR, pol, gag, and env coding regions. Fourteen patients with definite MS, 14 disease controls, 12 normal controls, and 3 patients with HTLV-I-associated myelopathy (HAM) were investigated. Results of particle aggregation assay for HTLV-I antibodies were negative in serum from all subjects except for the 3 HAM patients. Neither the 14 MS patients nor the 26 controls showed the presence of any highly homologous sequences to HTLV-I. We did observe faint signals for gag, pol, and env coding regions only at low stringent hybridization in some MS patients as well as some normal controls. The nucleotide sequence analysis of the faint bands was more homologous to major histocompatibility complex molecules than the HTLV-I genome.     

Intraluminal thrombosis and growth-mechanism of giant intracranial aneurysms

12 giant intracranial aneurysms were studied by MRI. Intraluminal thrombosis was observed in 9 aneurysms. Thrombosis was found more frequently in larger aneurysms. Thrombi were formed posteriorly or inferiorly in the lumen of 4 among 5 IC-cavernous aneurysms. Location of the neck of the aneurysms and stagnation of blood flow influenced by gravity may be causative factors determining the location of thrombi. In 6 aneurysms intraluminal thrombi were inhomogeneous on MRI, suggesting that the thrombi had been formed at different times. New thrombi were formed between the aneurysmal wall and the old thrombus in 3 cases. Dissection of the aneurysmal wall by residual blood flow in the lumen or hemorrhage in the aneurysmal wall may be one of the growth mechanisms of giant intracranial aneurysms.     

Characteristics of rhabdomyosarcoma cell lines derived from uterine carcinosarcomas

 Rhabdomyosarcoma (RMS) is occasionally found in the female genital tract, and mostly appears as one of the heterologous mesenchymal components in uterine carcinosarcoma designated as malignant mixed müllerian tumour (MMMT). We examined the biological properties of a pure rhabdomyosarcoma (RMS) cell line designated FU-MMT-3, which was newly established from a surgical specimen taken from a patient with uterine MMMT. We also evaluated c-myc and MYCN gene amplification in three RMS cell lines (including FU-MMT-3) derived from three MMMTs by Southern blot analysis. FU-MMT-3 cells were propagated continuously for 57 serial passages over a 2-year period in vitro. FU-MMT-3 was able to produce tumours demonstrating pure RMS in athymic nude mice. Cytogenetically, FU-MMT-3 showed a triploidy pattern, with complex karyotypic abnormalities including trisomy of chromosome 8. All three RMS cell lines, including FU-MMT-3, showed amplification of the c-myc gene (approximately fourfold to eightfold), while no cell lines demonstrated MYCN gene amplification. FU-MMT-3 is considered to provide a useful system for the study of the biological behaviour of RMS in MMMTs. Extra copies of chromosome 8 and c-myc gene amplification may be associated with the rhabdomyoblastic differentiation in MMMT. Received: 7 January 1997 / Accepted: 2 May 1997     

Neurogenic bladder after operation of rectal cancer

Of the patients who had had a urodynamic examination during the five year period from 1982 to 1987 in our clinic, 48 patients underwent operations for rectal cancer prior to the study. In 35 of them, the operation mode was known. If the pelvic nerve is damaged by operative modes for rectal cancer, urinary disturbances of severe kinds may occur. In spite of such disturbances, 71.4% of those who had had excision of the low anterior part and 51.9% of those with Miles' operation could be weaned from the clean intermittent self-catheterization and take up spontaneous urination. Even in patients who developed severe dysuria, if catheterized at an early stage, many of them could urinate by abdominal pressure with in several months after operation, without the aid of a catheter. This transition took place mostly within one year after operation. When a patient develops dysuria after radical surgery for rectum cancer, treatment mainly with self-catheterization is an effective method at present.     

Interstitial deletion of the short arm of chromosome 6 as a new cytogenetic marker of T-cell lymphoma

Three patients with malignant lymphoma, two of T-cell type and the other probably also of T-cell type, had an interstitial deletion of the short arm of chromosome 6, del (6)(p21p23), in common in their lymphoma cells. These findings suggest that this new type of chromosome deletion may be a specific marker for T-cell lymphoma and may play an important role in the lymphomagenesis and the expression of the T-cell phenotype.     

Ferritin immunohistochemistry as a marker for microglia

Summary An immunohistochemical analysis of formalin-fixed, paraffin-embedded brain sections was performed with antisera against holoferritin and the light(L)-subunit of ferritin. Sections immunostained using anti-glial fibrillary acidic protein (GFAP), Ricinus communis agglutinin-1 (RCA-1) stain for microglia and iron stain (Berlin blue stain) were compared. The L-subunit of ferritin was purified from normal human spleen according to the modified scrapie-associated fibrils purification, and the antiserum was raised in a rabbit. Both ferritin antisera positively stained resting and, more markedly, reactive microglia, both of which were also stained with RCA-1 but not with GFAP. Ferritin-positive resting microglia were seen more abundantly in cerebral and cerebellar cortices than in white matter. The advantages of ferritin antisera over RCA-1 are as follows. (1) RCA-1 heavily stains blood vessels, while anti-ferritin does not, hence the microglial cells are more readily visualized with ferritin immunohistochemistry. (2) Reactive microglia and macrophages are more strongly stained with anti-ferritin. (3) The staining intensity of ferritin is independent of the length of tissue fixation in formalin. However, anti-ferritin is inferior to RCA-1 in staining resting microglia with a scanty cytoplasm, especially in the white matter, probably because the former recognizes cytoplasmic components, while the latter recognizes cell membrane. Iron stain only gave a reaction to microglial cells in brains with neurosyphilis and to hemosiderin-laden macrophages. Thus, in addition to RCA-1, ferritin antisera are useful as a microglia marker in formalin-fixed, paraffin-embedded sections.Supported in part by Dr. A. Kondo, Department of Neuropathology, Neurological Institute, Kyushu University     

TECHNIQUES AND PITFALLS OF ENDOSCOPIC SUBMUCOSAL DISSECTION FOR COLORECTAL TUMORS

Endoscopic submucosal dissection (ESD) for colorectal tumors is steadily being developed. Safety and standardization of ESD for colorectal tumors have not been yet established because of the technical difficulties and the unsuitable anatomical characteristics of the colon and rectum. The authors mainly use a Flex knife for mucosal incision and a Hook knife for submucosal dissection to perform ESD safely. Skillful colonoscopic control, selection of scope, distal attachment tip hood, adequate high‐frequency generator and correct approach strategy should all be considered for safe performance of ESD. However, the incidence of indicative lesions is rare because the majority of colorectal tumors are adenomatous large laterally spreading tumors, which can be cured by intentional endoscopic piecemeal resection. At present, ESD for colorectal tumors should be performed only at central facilities that have expert colonoscopists. With the development of new devices and associated techniques, technical standardization of ESD for colorectal tumors is expected in the near future.     

Enzymes induced by ethanol differently affect the pharmacokinetics of trichloroethylene and 1,1,1-trichloroethane.

This study was undertaken to clarify the effect of enzymes induced by ethanol consumption on the pharmacokinetics of trichloroethylene (TRI, a highly metabolised substance) and 1,1,1-trichloroethane (1,1,1-TRI, a poorly metabolised substance). Rats maintained on a control liquid diet or a liquid diet containing ethanol (2 g/day/rat) for not less than three weeks were exposed to either TRI (50, 100, 500, and 1000 ppm) or 1,1,1-TRI (50, 100, and 500 ppm) by inhalation for six hours and the concentration of each compound in the blood and the urinary excretion of metabolites (trichloroethanol and trichloroacetic acid) were measured over several hours. Ethanol, which increased the in vitro metabolism of both compounds about fivefold, enhanced the in vivo metabolism of TRI only at high levels of exposure (marginally at 500 and considerably at 1000 ppm), whereas the metabolism of 1,1,1-TRI was enhanced at all concentrations tested. Moreover, there was a definite difference in the effect of induction of enzymes between the two solvents: the enhanced metabolism of TRI in vivo was shown by a decrease in the blood concentration of TRI as well as by an increase in the urinary excretion of its metabolites, whereas that of 1,1,1-TRI was shown by an increase in the urinary excretion of its metabolites alone. These results suggest that the induction of enzymes differentially affects the pharmacokinetics of TRI and 1,1,1-TRI in human occupational exposure: TRI metabolism may be increased only at concentrations much higher than the current occupational exposure limit (mostly 50 ppm), whereas 1,1,1-TRI metabolism may be increased at an exposure similar to occupational exposure.     

Characteristic findings of hepatocellular carcinoma: an evaluation with comparative study of US, CT, and MRI.

The sensitivity, specificity, and accuracy of ultrasonography (US), dynamic incremented computed tomography (CT) with delayed phase imaging, and magnetic resonance imaging (MRI) with or without Gd-DTPA were studied for detecting the characteristic appearances of hepatocellular carcinomas (HCC): fibrous capsules, fibrous septa, and mosaic appearances. Results were prospectively evaluated in 30 patients who subsequently underwent hepatic lobectomies or segmentectomies. Pathologic evaluations of the resected liver specimens demonstrated fibrous capsules in 20 tumors (66.7%), fibrous septa in 13 tumors (43.3%), and mosaic appearances in 19 tumors (63.3%). The accuracies for fibrous capsules were 71.4% (20 of 28) for US, 81.5% (22 of 27) for CT, and 92.3% (24 of 26) for MRI. The accuracies for fibrous septa were 57.1% (16 of 28) for US, 59.3% (16 of 27) for CT, and 73.1% (19 of 26) for MRI. The accuracies for mosaic appearances were 71.4% (20 of 28) for US, 51.9% (14 of 27) for CT, and 69.2% (18 of 26) for MRI. Gd-DTPA administered MRI showed higher accuracies than did conventional MRI for all manifestations. In conclusion, the fibrous capsules of HCCs were readily detected by CT and MRI. Gd-DTPA administration demonstrated an advantage in clarifying fibrous capsules, as well as fibrous septa and mosaic appearances.