Correlation among lung damage after radiation, amount of lipid peroxides, and antioxidant enzyme activities

The correlation between lipid peroxidation and morphologic changes was examined in Sprague-Dawley rat lungs after 30 Gy single thoracic radiation. The rats were sacrificed every week until the end of the fifth week after radiation. The left lungs were used for the measurement of lipid peroxides and antioxidant enzymes activities. The right lungs were examined by light and electron microscopy. Amounts of lung lipid peroxides were within normal limits, and no cellular degenerative changes were observed in the lungs except for subendothelial and interstitial edema 2 weeks after radiation. Lipid peroxides drastically increased and marked degenerative cellular changes such as edematous swelling, vacuolation, and destruction of cell membranes occurred in the alveolar septa following the third week after radiation. The activities of catalase were significantly higher during the period from the second to the fifth week and those of superoxide dismutase and glutathione peroxidase increased at the end of the fifth week. Our results demonstrated that the acceleration of lipid peroxidation was well correlated with the morphologic expression of cell injury in the irradiated lungs.     

Relationship between presence of Creola bodies and airway hyperresponsiveness in patients with bronchial asthma

It has recently been suggested that epithelial damage participates in the development of bronchial hyperresponsiveness. In this study we investigated the relationship between the presence of clusters of desquamated respiratory epithelial cells (Creola body. CrB) in sputum and airway responsiveness. Sputa were collected from asthmatic patients and acetylcholine inhalation tests were performed to assess airway responsiveness. Smears of 100 microliters of sputum were spread in pairs of two glass slides. One slide was stained with Papanicolaou's stain, the other with Giemsa stain. CrBs were detected on the whole glass slide stained with Papanicolaou's stain and the CrB score was determined by summing up the points given to each CrB on a glass slide according to the number of epithelial cells composing one CrB. The CrB score was significantly higher in patients with CrBs than in patients without CrBs. The patients with higher CrB score (CrB score greater than or equal to 6) had significantly greater number of eosinophils in sputa than the patients without CrBs. These results suggest the participation of damage to the respiratory epithelium in the development of airway hyperresponsiveness in bronchial asthma. The detection of CrBs is easy and we consider it clinically useful in the estimation of airway hyperresponsiveness.     

Muddy lung

A 31-year-old man, a racing car driver, was submerged in muddy water as the result of an accident. He died from respiratory failure after a 17-day clinical course. Foreign body granulomatosis and massive fibrosis of the lung were revealed at autopsy. The crystalline foreign bodies mainly were composed of silicon and ranged in size from 20 micron to 500 micron in diameter (average, 90 micron). Their distribution in the lungs corresponded to the areas of lung carnification. In this study, the authors demonstrate that near drowning in muddy water causes pulmonary silicate granulomatosis associated with carnificating fibrosis of the lung and term the pulmonary changes "muddy lung."     

Increased concentrations of secretory leukocyte protease inhibitor in peritoneal fluid of women with endometriosis

Secretory leukocyte protease inhibitor (SLPI) is a potent inhibitor of human leukocyte elastase. We investigated whether SLPI was present in the peritoneal fluid of women with endometriosis and to clarify the role of SLPI in the pathogenesis of endometriosis. Western blot analyses revealed that SLPI protein was detected as a 12 kDa band in peritoneal fluid. The peritoneal fluid concentrations of SLPI, elastase and interleukin-6 were assayed by enzyme-linked immunosorbent assays (ELISA). SLPI concentrations and the SLPI/elastase ratio in the peritoneal fluid of women with endometriosis were higher than in samples from women without endometriosis. There was no significant correlation between concentrations of SLPI and interleukin-6 in the peritoneal fluid. Immunohistochemistry using an anti-SLPI polyclonal antibody revealed positive staining in peritoneal macrophages, but not lymphocytes. The present findings suggest that SLPI found in the peritoneal fluid of patients with endometriosis may contribute to the pathogenesis of endometriosis.     

Effects of recombinant cytokines on murine megakaryocyte colony formation in a serum-free fibrin clot culture system.

A convenient serum-free fibrin clot culture system for murine megakaryocyte progenitor cells was developed. The culture and counting of colonies is much easier in this system, when compared with previously reported serum-free culture methods. Recombinant murine interleukin-3 (rmIL-3) stimulated megakaryocyte colony formation in a dose-dependent manner in this system. While recombinant human granulocyte colony-stimulating factor (rhG-CSF) had no effect on megakaryocytopoiesis, recombinant human erythropoietin (rhEpo) and recombinant human interleukin-6 (rhIL-6) augmented megakaryocyte colony formation stimulated by rmIL-3. The depletion of adherent cells and T cells from the cultured bone marrow did not eliminate the synergistic effect of rhEpo and rhIL-6.     

Multispacer typing of Rickettsia prowazekii enabling epidemiological studies of epidemic typhus

Currently, there is no tool for typing Rickettsia prowazekii, the causative agent of epidemic typhus, currently considered a potential bioterrorism agent, at the strain level. To test if the multispacer typing (MST) method could differentiate strains of R. prowazekii, we amplified and sequenced the 25 most variable intergenic spacers between the R. prowazekii and R. conorii genomes in five strains and 10 body louse amplicons of R. prowazekii from various geographic origins. Two intergenic spacers, i.e., rpmE/tRNA(fMet) and serS/virB4, were variable among tested R. prowazekii isolates and allowed identification of three and two genotypes, respectively. When the genotypes obtained from the two spacers were combined, we identified four different genotypes. MST demonstrated that several R. prowazekii strains circulated in human body lice during an outbreak of epidemic typhus in Burundi. This may help to discriminate between natural and intentional outbreaks. Our study supports the usefulness of MST as a versatile method for rickettsial strain genotyping.     

The Serum Factor from Patients with Ulcerative Colitis that Induces T Cell Proliferation in the Mouse Thymus Is Interleukin-7

The disturbance of immune regulatory T cells is related to the pathogenesis of ulcerative colitis. Here we demonstrated and characterized the serum factor from ulcerative colitis patients that induced proliferation of intrathymic T cells. The factor isolated from the patient sera by a combination of gel filtration and anion-exchange chromatography induced proliferation of CD4⁺CD8^– intrathymic T cells in the organ-cultured embryonic mouse thymus. Purification and amino acid sequence analysis of the serum factor demonstrated that the N-terminal 12 sequence was homologous to that of interleukin-7. SDS-PAGE and Western blot confirmed that purified serum factor was interleukin-7. Enzyme immunoassay demonstrated that the serum interleukin-7 concentration was significantly increased in the patients. PCR and Southern blot hybridization demonstrated that interleukin-7 mRNA expression was increased in the thymus tissues from patients but decreased in the colonic mucosa. Since interleukin-7 is a crucial cytokine for proliferation and differentiation of T cells in the thymus, the present study indicates that interleukin-7 may contribute to the disturbance of immune regulatory T cells in ulcerative colitis.     

On the various manifestations of spontaneous autoimmune diabetes in rodent models

Autoimmune (type 1) diabetes mellitus in mouse, rat, and humans shares several features, including T lymphocyte infiltration into pancreatic islets and a dependence on permissive class II major histocompatibility complex (MHC) alleles. We report here on an experimental model involving mice that express influenza hemagglutinin (HA) under the control of the insulin promoter and, at the same time, a transgenic class II MHC-restricted T cell receptor (TcR) specific for an HA peptide. These mice spontaneously develop islet infiltrates resembling those found in NOD mice and most animals become diabetic within 8 weeks of age. Because of the availability of a clonotypic TcR antibody, we can be confident that the Ins-HA transgene does not induce any measurable alterations in the vast majority of T cells with the transgenic TcR in primary and secondary lymphoid organs. Continuous export of large numbers of HA-specific lymphocytes from the thymus was not required for the manifestation of the disease since mice thymectomized at 3 days after birth still developed the disease albeit with smaller infiltrates.     

Long‐term observation of a Japanese mucolipidosis IV patient with a novel homozygous p.F313del variant of MCOLN1

Mucolipidosis type IV (MLIV) is an autosomal recessively inherited lysosomal storage disorder characterized by progressive psychomotor delay and retinal degeneration that is associated with biallelic variants in the MCOLN1 gene. The gene, which is expressed in late endosomes and lysosomes of various tissue cells, encodes the transient receptor potential channel mucolipin 1 consisting of six transmembrane domains. Here, we described 14‐year follow‐up observation of a 4‐year‐old Japanese male MLIV patient with a novel homozygous in‐frame deletion variant p.(F313del), which was identified by whole‐exome sequencing analysis. Neurological examination revealed progressive psychomotor delay, and atrophy of the corpus callosum and cerebellum was observed on brain magnetic resonance images. Ophthalmologically, corneal clouding has remained unchanged during the follow‐up period, whereas optic nerve pallor and retinal degenerative changes exhibited progressive disease courses. Light‐adapted electroretinography was non‐recordable. Transmission electron microscopy of granulocytes revealed characteristic concentric multiple lamellar structures and an electron‐dense inclusion in lysosomes. The in‐frame deletion variant was located within the second transmembrane domain, which is of putative functional importance for channel properties.