Surfactant protein C G100S mutation causes familial pulmonary fibrosis in Japanese kindred.

Several mutations in the surfactant protein C (SP-C) gene (SFTPC) have been reported as causing familial pulmonary fibrosis (FPF). However, the genetic background and clinical features of FPF are still not fully understood. We identified one Japanese kindred, in which at least six individuals over three generations were diagnosed with pulmonary fibrosis. We examined the patients radiologically and histopathologically and sequenced their SFTPC and ABCA3 genes. We also established a cell line stably expressing the mutant gene. All the patients had similar radiological and histopathological characteristics. Their histopathological pattern was that of usual interstitial pneumonia, showing numerous fibroblastic foci even in areas without abnormal radiological findings on chest high-resolution computed tomography. No child had respiratory symptoms in the kindred. Sequencing of SFTPC showed a novel heterozygous mutation, c.298G>A (G100S), in the BRICHOS domain of proSP-C, which co-segregated with the disease. However, in the ABCA3 gene, no mutation was found. In vitro expression of the mutant gene revealed that several endoplasmic reticulum stress-related proteins were strongly expressed. The mutation increases endoplasmic reticulum stress and induces apoptotic cell death compared with wild-type SP-C in alveolar type II cells, supporting the significance of this mutation in the pathogenesis of pulmonary fibrosis.

Increased prevalence of Kaposi΄s sarcoma-associated herpesvirus in the Kaposi΄s sarcoma-endemic area of western Kenya in 1981-2000.

Kaposi΄s sarcoma (KS) had been endemic in Africa before the appearance of human immunodeficiency viruses (HIV) in 1985. Incidence of African KS has increased over the time and the risk of contracting KS become greater in HIV-positive as opposed to HIV-negative individuals. KS specimens were collected in 1981-2000 from 228 surgical cases originating from a KS-endemic area of Western Kenya and examined for Kaposi΄s sarcoma-associated herpesvirus (KSHV) by an immunoperoxidase assay. The results showed that the specimens from 1981-1985 (before the HIV epidemic) were KSHV-positive in 10.3% in contrast to the KSHV positivity of 50.1-63.5% in 1986-2000. The linear increase of KSHV positivity in 1981-2000 was statistically significant. The most plausible explanation for the increased prevalence of KSHV in KS cases is that the endemic KS has changed to the epidemic one.

Detection of Human papillomavirus and cellular regulators p16INK4a, p53, and NF-kappaB in penile cancer cases in Kenya.

Human papillomavirus (HPV) E6 and E7 gene products play a central role in the induction of benign proliferation and malignant transformation by interacting with several cellular regulatory proteins such as p53, p16(INK4a), and nuclear factor kappaB (NF-kappaB). In this study, HPV DNA was detected by in situ hybridization (ISH) and p53, p16(INK4a), and NF-kappaB by immunochemistry in 22 penile cancer cases in Kenya. HPV DNA was found in 68.2% of the cases. There was no difference in the p53- and p16(INK4a)-positivities in HPV DNA-positive and HPV DNA-negative cases. In HPV DNA-positive cases, the NF-kappaB positivity in the nucleus, cytoplasm, and nucleus and/or cytoplasm amounted to 73.3%, 93.3%, and 100%, respectively, while in HPV DNA-negative cases, a 28.7% NF-kappaB positivity of in the nucleus and/or cytoplasm was observed. It is concluded that NF-kappaB in penile cancer is expressed more frequently in the presence of HPV infection than in its absence.

Pluripotent hemopoietic precursors in vitro (CFUMIX) in aplastic anemia.

When human marrow cells were cultured in a medium containing alpha-medium, methylcellulose, fetal calf serum, bovine serum albumin, erythropoietin, and leucocyte-conditioned medium, mixed colonies composed of erythrocytic cells and granulocytes were formed. The clonal nature of the mixed colonies was confirmed by the linear relationship between the numbers of cells plated and the number of colonies, and the absence or presence of Y-chromatin in the mixed colonies in a co-culture experiment with male and female cells. Using the methylcellulose cell culture techniques, the pluripotent hemopoietic precursors (CFUMIX) in marrow cells from 15 patients with aplastic anemia were assayed. In the control subjects of patients with iron-deficiency anemia, lymphoadenitis, reactive leucocytosis or Hodgkin's disease, 8 X 10(5) marrow cells in 4 dishes produced 12.7 +/- 6.9 (mean +/- SD) mixed colonies. On the other hand, 8 X 10(5) marrow cells from patients with aplastic anemia formed only 2.1 +/- 5.5 (mean +/- SD) mixed colonies. Furthermore, the marrow cells from 5 patients who were repeatedly receiving transfusions contained no CFUMIX which give rise to mixed colonies. The present results provided the first direct evidence that pancytopenia in most patients with aplastic anemia results from a reduced influx into the compartment of maturing hemopoietic cells from the compartment of pluripotent hemopoietic precursors.

Eosinophilic globules in pheochromocytoma of the adrenal medulla. A histochemical, immunohistochemical and ultrastructural study.

Histochemical, immunohistochemical, and ultrastructural characteristics of eosinophilic globules in pheochromocytoma of the adrenal medulla are described. The globules were observed in 7 (63.5%) out of 11 cases of pheochromocytoma of the adrenal medulla. These globules were eosinophilic, PAS-positive with and without diastase predigestion, phosphotungstic acid hematoxylin (PTAH) positive, and autofluorescent under ultraviolet illumination. Although almost all globules were not stained with neuron specific enolase (NSE) using the immunoperoxidase method, a few globules were stained positive. The lectins of Triticum vulgaris (WGA) and Ricinus communis (RCA-120) were weakly bound to most of the eosinophilic globules. Immunohistochemical reactions for alpha-fetoprotein (AFP), alpha-l-antichymotrypsin (ACT), and human chorionic gonadotropin (HCG) revealed negative reaction in these globules. An ultrastructural study revealed no relationship between these globules and chromaffin secretory granules. These findings suggested that the eosinophilic globules in pheochromocytoma of the adrenal medulla were not related to the chromaffin secretory granules and might be some kind of complex protein.

Staining method for hepatitis B surface antigen (HBs Ag) and its mechanism.

HBs Ag in the hepatocytes has been stained by the orcein method. As degenerative and necrotic cytoplasms of hepatocytes are occasionally stained also by this method, it is difficult to identify HBs Ag. The orcein dye has different staining properties from lot to lot, causing difficulty in obtaining stable results. Therefore, the author tried to modify the orcein method and found that the procedure using a sensitizer (ferric ammonium sulfate or uranium nitrate) after oxidizing the solution gave stable and satisfactory results. By this method, in the hepatocytes, only HBs Ag is stained positively. The mechanism of function of the sensitizer is not clear, but it is considered that ferric or uranium ions occupy small gaps in the protein molecular structure in the cells before orcein molecules occupy these gaps and that the method does not give any false positive result. It is also suggested that various HBs Ag staining dyes can react with sulfonic acid residue groups which are formed by the oxidization of disulfide group of proteins. Another new staining method for HBs Ag using resorcin fuchsin has been developed. The results of this method were also discussed.

Statistical analysis of relationship between iron accumulation and hepatitis B surface antigen.

Statistical analysis was carried out to determine the relationships of iron deposition among hepatocytes, Kupffer cells, bile duct epithelial cells, portal tracts, and spleens using 68 autopsy cases obtained from Kenya, which included cases of Bantu siderosis. Of 68 cases, 33 (49%) showed siderosis. Cirrhosis was observed in 19 cases, of which 10 (53%) showed siderosis. However, there was no correlation between iron deposition and cirrhosis (Pearson chi 2 = 0.6734). The finding suggests that iron accumulation does not lead to cellular injury. On the other hand, there was a strong association between iron deposition in the livers and that in the spleens. There was also a strong correlation between the presence of hepatitis B surface antigen and iron deposition in the Kupffer cells and in the spleens. Therefore, it is suggested that the hepatitis B virus infection affects the iron metabolism of the reticuloendothelial system.

Long-term trend of decreasing frequency of hepatitis B virus infection.

Statistical and histopathological analysis was carried out to determine whether hepatitis B surface antigen (HBsAg) with the autopsy diagnosis of hepatitis, cirrhosis, and hepatocellular carcinoma declined during 1964-1991. In this study, the liver specimens from 673 autopsy cases at Nagasaki University Hospital were used, and these materials were stained for HBsAg with immunoperoxidase method. For statistical calculation, we used Cochran's chi-square test with one degree of freedom for linear trend for proportions. We investigated HBsAg positive rates for the four time periods, 1964-1970, 1971-1977, 1978-1984, and 1985-1991, and for the diagnosis of hepatitis, cirrhosis, and hepatocellular carcinoma. The decreasing linear trend of HBsAg positivity with time was significant for hepatocellular carcinoma and cirrhosis.

Endocarditis caused by Candida parapsilosis.

The authors report a case of endocarditis caused by Candida parapsilosis. To the best of our knowledge, a case has not been described previously in Japan in the English literature. A battery of 8 peroxidase-labeled lectins was tested on sections of paraffin-embedded tissue to determine which lectin could be used in the microscopic diagnosis of C. parapsilosis. One lectin, from Archis hypoaea (PNA) was found to react with C. parapsilosis. On the other hand, C. albicans, Aspergillus, Mucor, and Cryptococcus did not react with A. hypoaea (PNA). On fluorescence microscopic study, C. parapsilosis was not fluorescent, but other fungi were fluorescent when exposed to ultraviolet illumination. Therefore, we propose new procedures for identification of C. parapsilosis in tissue sections using lectin histochemistry and fluorescence microscopy.

[A fatal case of streptococcal toxic shock-like syndrome probably caused by acupuncture].

A 41-year-old male received acupuncture in the right shoulder for the sake of arthralgia. Three days after acupuncture he was admitted due to severe epigastralgia. Erythematous change and swelling were observed around the right shoulder. A study by magnetic resonance showed an increased signal intensity in a portion of the right subscapular muscle. Four hours after admission he became hypotensive. The erythematous and necrotic change in the right shoulder skin rapidly spread. Excisional debridement in the right lateral chest wall was immediately done. However, the patient died one day after admission despite administration of a high-dose ampicillin and other supportive therapies. Bacteriological and histological examinations confirmed severe streptococcal myositis. This is a case report of toxic shock-like syndrome probably caused by acupuncture.

Immunohistochemical reactivity of phagocytic and non-phagocytic histiocytes in lymph nodes with lysozyme, alpha-1-antichymotrypsin, S-100 protein, alkaline phosphatase, and acid phosphatase.

Yellow-brown bodies were observed in the sinusoids of lymph node and histiocytes. The authors confirmed immunohistochemical reactivity of lysozyme, alpha-1-antichymotrypsin, S-100 protein, alkaline phosphatase, and acid phosphatase in non-phagocytic and phagocytic histiocytes which contained yellow-brown bodies. Phagocytic histiocytes (histiocytes with yellow-brown bodies) were not reacted with lysozyme, alpha-1-antichymotrypsin, S-100 protein, alkaline phosphatase, and acid phosphatase. On the other hand, non-phagocytic histiocytes were reacted with lysozyme, alpha-1-antichymotrypsin, S-100 protein, alkaline phosphatase, and acid phosphatase.

[A case report of giant cell granuloma of the jaw].

A giant cell granuloma of the mandible seen in a 16 month-old boy was reported. Histopathology showed that the lesion consisted basically of granulation tissue with multinucleated giant cells, spindle-shaped and polygonal stromal cells. No atypia was seen in giant cells or stromal cells. Electron microscopic examination revealed that multinucleated giant cells possessed microvillous structure on their cytoplasm and were rich in organelles such as endoplasmic reticulum, mitochondria and dense bodies. Stromal cells consisted of macrophage-like and fibroblastic cells. Immunohistochemical examination demonstrated a large number of giant cells and stromal cells that were strongly positive for the antibodies of alpha-1-antitrypsin and alpha-1-antichymotrypsin. These morphological findings suggest that macrophage-like stromal cells are the precursors to giant cells.

[Kaposi's sarcoma].

Kaposi's sarcoma (KS) (Kaposi, 1872) is classified into at least three forms: 1) the classic form; in elderly men of southern European, Jewish, and Italian origin, 2) the endemic form; among native populations in equatorial Africa and 3) the epidemic form; as a complication of AIDS. As one additional form could be posttransplantation KS in organ transplant recipients. Histologically, KS is composed of two characteristic features; a proliferation of angiomatous lesion and of spindle-shaped cells. The pathological entity of the disease is still controversial. As a cause of the disease, human cytomegalovirus (HCMV) has been the subject based on epidemiologic, serologic, geopathologic and histopathologic observations. About half of KS cases with AIDS has shown evidence of HCMV infection as cytomegalic viral inclusions of the cells. In contrast, about 30% of AIDS cases without KS showed histological HCMV infection. A connection between HCMV and KS was suggested by the detection of herpes-type virus particles in several KS tissue culture lines and in a tumor specimen. DNA and RNA homologous with HCMV sequences have been detected in tumor tissue of both endemic and epidemic KS by electrophoretic methods. HCMV sequences was detected in extracted DNA of 30% of classic KS, using Southern blot technique. In situ hybridization technique using specific DNA probes for HCMV has been developed to reveal endogenous nucleic acid sequences in fixed tissue sections. However, some reports showed that by the in situ technique only a small number of KS cells in the AIDS cases and no case of the classic KS were positive for HCMV within KS lesions. HCMV infections are ubiquitous. There is a very high seropositivity of HCMV in African inhabitants as well as in Africans with endemic KS. A strong association between HCMV and KS was not known. As for HIV itself in AIDS with KS, previous studies have showed the presence of chromosomal abnormalities of KS cells, but not revealed the presence of HIV DNA sequences in KS. Vascular tumors were produced by NIH/3T3 cells transfected with KS DNA of AIDS. An oncogene isolated by transfection of KS DNA encodes a growth factor; FGF. Isolation of rearranged human transforming gene following transfection of KS DNA was performed. DNA having transforming genetic elements which have been repetitively transmitted into phenotypically normal NIH/3T3 cells through cycles of transfection was isolated from tissues of KS with AIDS.(ABSTRACT TRUNCATED AT 400 WORDS)