A successfully treated case of mycetoma due to Nocardia veterana.

We report a successfully treated case of mycetoma from which an unusual Nocardia species was isolated. The isolate was identified as N. veterana by biochemical characterization and 16S ribosomal RNA gene sequencing, and it has not been previously reported as a causative agent of human mycetomas. Treatment with various antibiotics over 6 years and surgical resection failed to cure the disease. However, the combination of intravenous imipenem/cilastatin and amikacin along with oral clarithromycin and minocycline proved very effective in this case. This is the first case report of mycetoma due to N. veterana in a clinical setting.

A novel GGNG-related neuropeptide from the polychaete Perinereis vancaurica.

The GGNG peptides are myoactive peptides so far identified from earthworms and leeches, which are the earthworm excitatory peptides (EEP) and the leech excitatory peptide (LEP), respectively. A novel GGNG peptide was isolated and structurally determined from a marine polychaete, Perinereis vancaurica, using a combination of immunological assay and high performance liquid chromatography (HPLC). The peptide was a pentadecapeptide whose amino acid sequence was similar to that of EEP and LEP, and showed myoactivity on isolated esophagus of P. vancaurica with a threshold concentration of 10(-10)M. The peptide was designated as polychaete excitatory peptide (PEP). Amidation of the alpha-carboxyl group of C-terminal residue occurred in PEP. This is the case for LEP, but not for EEP. The cDNA cloning revealed that the structure of the PEP precursor is more similar to the EEP precursor than to the LEP precursor. Immunohistochemical staining showed the presence of PEP in several neurons of central nervous system (CNS) as somata and neuropile structure, epithelial cells of the pharynx and epidermal cells throughout the body wall. Altogether these results support the physiological significance of PEP in regulation of the CNS neural activity and the peripheral myoactivity.

A case of anaphylaxis due to ibuprofen.

A forty-four-year-old Japanese female, who had persistant rhinorrhea, was administered Benza block tablets orally along with two other medicines. Immediately after ingestion, the patient displayed itching of the right upper eyelid, followed by coughing, sneezing, nasal discharge, nasal obstruction, nausea, vomiting, swelling of the face, and dyspnea. She had edema, a wheal extending from the face to the neck, and swelling of the eyelids and lips. Her symptoms subsided after treatment. Her reaction to ibuprofen, which was contained in the Benza Block tablets, was confirmed by a positive reaction to prick testing. From the results of these examinations, our patient was diagnosed as having anaphylaxis due to the ibuprofen in the Benza Block tablets. A review of the literature revealed no previous reports of anaphylaxis due to ibuprofen, although a few cases of ibuprofen urticaria have been reported.

Possible functions of oxytocin/vasopressin-superfamily peptides in annelids with special reference to reproduction and osmoregulation.

Annetocin is an earthworm oxytocin-related peptide that we previously isolated from the whole body of a lumbricid earthworm Eisenia foetida. We have reported that annetocin induces egg-laying-like behaviors in E. foetida and a gnathobdellid leech, Whitmania pigra, when it is injected into the respective animals. The present study was undertaken to probe physiological functions of invertebrate oxytocin-vasopressin-superfamily peptides with special reference to reproductive and osmoregulatory events in which vertebrate peptides of this superfamily are involved. Annetocin, Lys-conopressin (a leech vasopressin-related peptide) and two analog peptides, [Tyr(3)]-annetocin ((3)Y-annetocin) and [Phe(3)]-annetocin ((3)F-annetocin), were compared for their activities to induce egg-laying-like behavior and to change body weight as a measure of water balance in the leech W. pigra. Injection of annetocin, Lys-conopressin, and (3)F-annetocin caused both egg-laying-like behavior and reduction of body weight in the animals, but (3)Y-annetocin induced neither. Furthermore, leeches in the non-breeding season responded to peptides less conspicuously than those in the breeding season. Such a concomitant induction of egg-laying-like behavior and body-weight reduction suggests that these two phenomena are unitary and might be accounted for by the fact that egg-laying in leeches and earthworms is accompanied by secretion of a large quantity of mucus, which should significantly contribute to body-weight loss. J. Exp. Zool. 284:401-406, 1999.

Light-regulated localization of the beta-subunit of Gq-type G-protein in the crayfish photoreceptors.

In crayfish photoreceptor cells, Gq-type G-protein plays a central role in the phototransduction pathway, and the translocation of Gq alpha has been proposed as one of the molecular mechanisms to control photoreceptor sensitivity. We here investigated beta subunit of Gq and its localization profiles under various light conditions in the crayfish photoreceptor cells to understand the functional characteristic of visual Gq in the phototransduction pathway. An immunoprecipitation experiment was performed using an anti-Gq alpha antibody and a thiol-cleavable crosslinker. A 39 kDa protein was co-immunoprecipitated with Gq alpha, but not by irradiation, in the presence of GTP gamma S. The partial amino acid sequence of the 39 kDa protein was similar to G beta e in Drosophila photoreceptors, indicating that the crayfish G beta which combines with Gq alpha is a G beta e homologue. Immunohistochemical and immunoblot analyses revealed that the amount of the G beta decreased in the rhabdomeric membranes and increased in the cytoplasm in the light, compared with that in the dark. The profile of the translocation was similar to that reported for Gq alpha. Since both alpha and beta gamma subunits are necessary for G-proteins to be activated by rhodopsin in the rhabdom, the light-modulated translocation of a G beta e homologue possibly controls the amount of Gq which can be activated by light-stimulated rhodopsin.

Immunohistochemical Localization of Annetocin, an Earthworm Oxytocin-Related Peptide, and Identification and Ultrastructural Characteristics of the Annetocin-Secretory Cells in the Oligochaete Earthworm Eisenia foetida.

Annetocin is an egg-laying-inducing oxytocin-related peptide which we have previously isolated from the earthworm, Eisenia foetida. Here we report the results of immunohistochemical and ultrastructural studies on annetocin-secretory cells in the earthworm. Annetocin-immunoreactive (IR) cell-somata were located mainly at the ventro-lateral side of the subesophageal ganglion. Only four annetocin-IR cells were seen in the cerebral ganglion. Some annetocin-IR cells displayed unipolar-like structure with a process directing to the core region (the neuropile) of the ganglion. Annetocin-IR fibers were also observed in the neuropile of the ventral ganglia and the ventral nerve cord between the 4th and the 30th segments including the clitellum, but not in the posterior segments (31-55th). The number of annetocin-IR fibers decreased from the 4th to the 30th segment. The annetocin-secretory cells were identified by the immunogold staining, and filled with gold-labeled vesicles, 200-250 nm in diameter, which included moderately electron dense material. The annetocin-secretory cells possessed a euchromatic nucleus, well-developed rough endoplasmic reticulum and Golgi apparatus. Some of the annetocin-secretory cells were found to form a neurohemal-like structure, where somata or fibers with loose glial investment came in contact with the coelomic space at the ventral side of the subesophageal ganglion. The results suggest that annetocin is a neuropeptide produced and secreted by the neuron in the cerebral and subesophageal ganglia of the earthworm.

Immunochemical demonstration of Eisenia tetradecapeptide, a bioactive peptide isolated from the gut of the earthworm Eisenia foetida, in tissues of the earthworm.

The quantity and localization of Eisenia tetradecapeptide which was isolated from the gut of the earthworm Eisenia foetida were examined in tissues of the same species by enzyme-linked immunosorbent assay and immunohistochemistry. Analysis by enzyme-linked immunosorbent assay showed that Eisenia-tetradecapeptide-like immunoreactivity was present in both the central nervous system (cerebral ganglion, subesophageal ganglion, ventral ganglia, and ventral nerve cord) and the gut (esophagus, crop, gizzard, and intestine). The central nervous system contained a higher amount of Eisenia-tetradecapeptide-like immunoreactivity (1.3 pmol/mg wet weight) than the gut (0.2-0.6 pmol/mg wet weight). Eisenia-tetradecapeptide-like immunoreactivity was scarcely detected in the body-wall muscle, nephridia, and sexual organs (testis, ovary, seminal vesicle, and ovisac). Immunohistochemical analysis demonstrated that intense Eisenia-tetradecapeptide-like immunopositive cells and nerve fibers were present in the central nervous system. Immunoreactivity was found in the epithelial cells lining the esophagus and in the submucous plexus in various parts of the gut. Thus, the present study suggests that Eisenia tetradecapeptide is a neuropeptide and/or peptide hormone present in both the central nervous system and the gut of the earthworm and that its role involves the regulation of gut motility.

IgE memory B cells identified by the polymerase chain reaction.

The existence of memory B cells for IgE was investigated using spleen cells harvested from mice primed and boosted with dinitrophenylated keyhole limpet hemocyanin. Using the polymerase chain reaction, it was found that mRNA for IgE was already demonstrable in spleen cells taken 1 day after boost. Spleen cells taken from mice 1 day after boost and cultured in vitro without the addition of antigen also produced anti-dinitrophenylated (DNP) IgE antibody. In contrast, mRNA for IgE in spleen cells from mice primed 4 weeks previously, but not boosted, was not demonstrable. Anti-DNP IgE was not produced in cultures of spleen cells from mice primed but not boosted. As mRNA for IgE was demonstrable already 1 day after boost we concluded that mice produce memory B cells not only for IgM, but also for IgE specific for the immunizing antigen.

Clonotypic analysis of T cells infiltrating the skin of patients with atopic dermatitis: evidence for antigen-driven accumulation of T cells.

Atopic dermatitis (AD) is a chronic inflammation of the skin characterized by marked infiltration of lymphocytes, suggesting an important role for cellular immune responses in the pathogenesis of AD. It is still unclear, however, whether accumulation of lymphocytes results from antigen-driven mechanisms or by nonspecific inflammatory processes. We applied a novel method of T cell clonotypic analysis, using a combination of reverse transcriptase-PCR with multiple T cell receptor (TCR) primers and subsequent single-strand conformation polymorphism (SSCP). Using this method, we were able to detect oligoclonal accumulation of T cells in inflamed skin areas of patients with AD, but not in normal skin. Identical T cell clones with the same antigen specificities were present in samples obtained from separate skin regions. Our results indicate that antigen(s) that exist in large areas of the skin stimulate and expand a relatively restricted number of antigen-specific T cells, leading to oligoclonal accumulation of T cells. We suggest that a marked antigen-driven infiltration of T cells is present in AD.

Light-modulated subcellular localization of the alpha-subunit of GTP-binding protein Gq in crayfish photoreceptors.

Gq-type GTP-binding protein (Gq) plays an important role in invertebrate visual phototransduction. The subcellular localization of the alpha subunit of visual Gq in crayfish photoreceptor was investigated immunocytochemically and biochemically to demonstrate the details of the rhodopsin-Gq interaction. The localization of Gq(alpha) changed depending on the light condition. In the dark, Gq(alpha) was localized in the whole rhabdoms as the membrane-bound form. In the light, half of the Gq(alpha) was localized in the cytoplasm as the soluble form. The translocation of Gq(alpha) was reversible. The light-modulated translocation possibly controls the amount of Gq that can be activated by rhodopsin. In vitro hydroxylamine treatment of rhabdomeric membranes suggested that the translocation was regulated by the fatty-acid modification of Gq(alpha).

Murine IgG1 and IgE memory B cells.

Anti-dinitrophenyl IgG1 and IgE antibody production were examined in vitro. Splenic B cells from mice immunized in vivo at various intervals with dinitrophenylated proteins were cultured in vitro with splenic T cells and splenic antigen-presenting cells from mice immunized previously (2 weeks) with keyhole limpet hemocyanin. Anti-dinitrophenyl IgG1 and IgE antibodies were determined in the supernatants by ELISA. Without T cells and antigen-presenting cells or without antigen, no anti-dinitrophenyl antibodies were detected. When the B cells were separated by panning with anti-murine IgG1 antibody, only the cells adhering to anti-IgG1 produced anti-dinitrophenyl IgG1 antibodies. Anti-dinitrophenyl IgE production was observed from cells adhering and from cells not adhering to the anti-IgG1-coated plates, as expected. However, anti-dinitrophenyl IgE was also produced from cells which adhered to the anti-IgG1-coated plates, but did not have surface IgE antibodies. Therefore, cells without surface IgE adhering to anti-IgG1 were capable of producing anti-dinitrophenyl IgE antibodies.

Granulocyte colony-stimulating factor (G-CSF) and granulocyte macrophage colony-stimulating factor (GM-CSF) in Behçet's disease.

Increases in the number and activity of peripheral polymorphonuclear neutrophils (PMNs) is often found in Behçet's disease (BD), indicating that PMN may play an important role in the pathogenesis of this disorder. It has recently been reported that G-CSF and GM-CSF, a family of hematopoietic growth factors, enhance PMN activity. To explore the role of these two CSFs in BD, we first examined the chemotactic response of PMNs to these CSFs by performing a polarization assay. PMN response to G-CSF in BD patients was lower than that in controls, while PMN response to GM-CSF was similar in patients and controls. However, PMNs from BD patients showed an enhanced chemotactic response to N-formyl-L-methionyl-leucyl-phenylalanine. Thus, it is speculated that the PMNs of the patients might have already been activated in vivo by G-CSF and thus could not respond further to this agent in vitro. We examined G-CSF and GM-CSF mRNA expressions in peripheral mononuclear cells stimulated with LPS, PMA, and Con A by Northern hybridization. G-CSF mRNA expression levels in BD patients were higher than in the controls, while GM-CSF mRNA expression levels were lower than in the controls. We also examined the serum levels of the two CSFs by ELISA and EIA. However, all levels of the two CSFs in both patients and controls were not detectable, except in the case of one BD patient in the active stage of the disease, who showed high levels of G-CSF, but not of GM-CSF.(ABSTRACT TRUNCATED AT 250 WORDS)

Evidence for a frontal-organ homologue in the pineal complex of the salamander, Hynobius dunni.

The pineal complex of larval and adult salamanders, Hynobius dunni, was examined by light and scanning electron microscopy. This pineal complex displays an anterior and a posterior portion, both of which possess a lumen. The anterior lumen is small and closed, whereas the posterior lumen is in open communication with the third ventricle. Cell processes of the photoreceptor cells and microvilli of the supportive cells are visible in both lumina. The anterior part of the complex is formed by an independent, second evagination from the common pineal anlage; this process takes place immediately after hatching. The anterior body of the pineal complex of H. dunni appears to be homologous to the frontal organ of anurans.

Structural and endocytotic differences of fibroblasts and macrophages in the tail fin of amphibian larvae during metamorphosis.

Different features in the fibroblasts and the macrophages, which are prominent cell types in the dermis of the dorsal tail fin of the larval axolotl, Ambystoma mexicanum, and the tadpole, Rana japonica, were examined by light and electron microscopy. At the non-metamorphic stages, the cytoplasm of the macrophage, loaded with numerous lysosomes, is generally located in the cell periphery. Outstanding was the presence of many ruffles or microvillous projections of different shapes and sizes in the plasma membrane. In contrast, the fibroblasts are spindle-shaped and possess less numerous microvillous projections compared with the macrophages, and extracellular spaces neighboring the fibroblasts are loaded with collagen fibers. The fibroblasts are located superficially and sometimes contact each other by the desmosomes. The macrophages are situated relatively deep in the dermis. At the metamorphic stages, both fibroblasts and macrophages contain many phagolysosomes in common, but the desmosomes still remain between the fibroblasts, and therefore the fibroblasts are distinguished from the macrophages. The macrophages are characterized by the their phagolysosomes which contain degenerating cells or cell debris of fibroblasts, myelinated nerve fibers and neutrophils. The macrophages also exhibit a higher endocytotic activity than the fibroblasts for the injected foreign particles (FITC-dextran or latex beads). On the other hand, the phagolysosomes (or autophagolysosomes) of the fibroblasts are characterized by intracellular collagen fibers. These different phagolysosomes between the macrophages and the fibroblasts mean that they may play different roles in the degeneration of the cellular or the extracellular components, respectively, of the tail fins during metamorphosis.

[A multicenter study on panipenem/betamipron in dermatology].

Panipenem/betamipron (PAPM/BP), a new carbapenem, was studied in dermatology. PAPM/BP was used clinically in the treatment of skin and skin structure infections in a multicenter trial. Fifty three patients were enrolled in the trial. Clinical evaluations were made in 50 patients. Most patients received intravenous infusion of PAPM/BP in a dose of 500 mg twice daily. Other dosages were used in some patients. The overall clinical efficacy rate was 78%. When 15 cases of secondary infections were excluded, the rate was 85.7%. Adverse responses were nausea and/or vomiting in 3 patients, redness with itching in 1 patient, headache or head heaviness in 2 patients and diarrhea in 1 patient. The patient with redness and itching had also nausea and vomiting. This occurred 1 hour after the start of the first infusion of this drug. After the discontinuation of the treatment the symptoms went away on the next day. Abnormalities in laboratory test results were observed in 7 out of 53 patients. One patient with liver cirrhosis and hepatocellular carcinoma developed anemia (RBC 372 x 10(4)/mm3----275 x 10(4)/mm3, Hb 11.9 g/dl----8.8 g/dl, 35.1%----26.0%). Other abnormalities were all mild. Penetration of the drug into skin tissues after intravenous infusion of 500 mg of this drug in skin surgery patients was studied. Skin/serum concentration ratios ranged from 0.20 to 0.97. Skin concentrations were higher than the concentration of PAPM inhibiting 80% of clinical isolates over a period of 6 hours. In rats, skin concentrations were much lower than serum concentrations probably due to the difference in in vivo metabolism of PAPM. A few resistant strains of Staphylococcus aureus against PAPM and imipenem (IPM) were isolated. However, PAPM and IPM showed good antibacterial activities compared to other drugs tested. In conclusion, PAPM/BP is considered to be a useful drug in the treatment of skin and skin structure infections.

Operations for microforms of cleft lip.

Microform cleft lip is a mild expression of cleft lip and may be difficult to repair. Its severity may be defined by the degree of downward depression of the nostril rim, skin striae of the upper lip, notching of Cupid's bow, and deformity of the vermilion border. Variation in surgical repair is reported for each type of microform cleft lip.