Bone metabolic microarray analysis of ligature-induced periodontitis in streptozotocin-induced diabetic mice.

BACKGROUND AND OBJECTIVE: Periodontal disease is a chronic infectious disease that results in bone loss. Many epidemiological studies have reported the progression of periodontal tissue destruction in patients with diabetes; however, the associated mechanism remains unclear. In this study, we comprehensively investigated how diabetes affects the periodontal tissue and alveolar bone loss using a ligature-induced periodontitis model in streptozotocin-induced diabetic (STZ) mice. MATERIAL AND METHODS: Diabetes was induced by intraperitoneal injection with streptozotocin in 6-wk-old C57/BL6J male mice. A silk ligature was tied around the maxillary left second molar in 9-wk-old wild-type (WT) and STZ mice. Bone loss was evaluated at 3 and 7 d after ligation. mRNA expression levels in the gingiva between the two groups were examined by DNA microarray and quantitative polymerase chain reaction at 1, 3 and 7 d post-ligation. Tartrate-resistant acid phosphatase and alkaline phosphatase staining of the periodontal tissue was performed for evaluation of osteoclasts and osteoblasts in histological analysis. RESULTS: In the gingiva, hyperglycemia upregulated the osteoprotegerin (Opg) mRNA expression and downregulated Osteocalcin mRNA expression. In the ligated gingiva, tumor necrosis factor-α (Tnf-α) mRNA expression was upregulated at 1 d post-ligation in STZ mice but not in WT mice. At 3 d post-ligation, alveolar bone loss was observed in STZ mice, but not in WT mice. Significantly severe alveolar bone loss was observed in STZ mice compared to WT mice at 7 d post-ligation. Bone metabolic analysis using DNA microarray showed significant downregulation in the mRNA expression of glioma-associated oncogene homologue 1 (Gli1) and collagen type VI alpha 1 (Col6a1) at the gingiva of the ligated site in STZ mice compared to that in WT mice. Quantitative polymerase chain reaction showed that Gli1 and Col6a1 mRNA expression levels were significantly downregulated in the gingiva of the ligated site in STZ mice compared to WT mice. Histological analysis showed lower alkaline phosphatase activity in STZ mice. In addition, an increased number of tartrate-resistant acid phosphatase-positive multinucleated cells were observed at the ligated sites in STZ mice. CONCLUSIONS: These results suggest that an imbalance of bone metabolism causes osteoclastosis in insulin-deficient diabetes, and that alveolar bone loss could occur at an early phase under this condition.

The first case of cutaneous mucormycosis caused by Rhizopus azygosporus.

A rapidly enlarging leg ulcer appeared in a 54-year-old woman with systemic lupus erythematosus receiving aggressive immunosuppressive therapy. Skin biopsy revealed proliferation of hyphae in the midst of a neutrophilic abscess. Culture yielded Rhizopus azygosporus. As no organ involvement was detected by thorough examination, the patient was diagnosed as having primary cutaneous mucormycosis. Although intravenous amphotericin B therapy seemed to be very effective, it had to be discontinued due to nephrotoxicity. She unfortunately died of subsequent disseminated fungal infection and cerebral infarction in which the primary cause could not be determined. Minimum inhibitory concentrations of several antifungal drugs to the isolate were examined and amphotericin B proved to be the only agent that may potentially reach the effective plasma concentration. This is the first case report of cutaneous mucormycosis caused by R. azygosporus.

Cutaneous zygomycosis: a case report and review of Japanese reports.

A 69-year-old man, a carpenter with idiopathic thrombopenic purpura and interstitial pneumonia, was treated with steroid pulse therapy and antibiotics. On the seventh day of steroid therapy, a conglomeration of papules, vesicles and pustules appeared in an area of the left buttock in contact with his napkin. In a Parker KOH specimen of the crust of the lesion, many non-septate hyphae were seen, and culture of material obtained by biopsy yielded Rhizopus microsporus var. microsporus. Ketoconazole cream was applied topically for 1 week, and the exanthema healed. After the third month of inpatient treatment, the patient's overall condition had returned to normal, and he was discharged. Cutaneous zygomycosis is a rare disease, and only 19 cases have been reported in Japan. Its characteristics, as reported in these cases, have been collected and collated.

Involvement of transcriptional repressor ATF3 in acceleration of caspase protease activation during DNA damaging agent-induced apoptosis.

DNA topoisomerase inhibitors are effective chemotherapeutic agents on several solid tumor cells. They induce a specific signaling cascade that executes an active cell death process (apoptosis), including caspase activation, and the blockage of the signaling is associated with drug-resistance of tumor cells. However, little is known about the initial signal transduction induced by the agents. In the present study, we screened genes that are initially upregulated in caspase-independent manner. We found that the activating transcription factor 3 (ATF3) protein, a repressor of cyclic-AMP responsive element (CRE)-dependent transcription, was strongly induced among CRE-BP/ATF members and subsequently accumulated in nuclei following camptothecin or etoposide treatment. During induction of apoptosis, the accumulation and the nuclear translocation of ATF3 coincided with the activation of caspase protease and were not inhibited by the broad caspase inhibitor Z-VAD-fmk, indicating that ATF3 induction is not a downstream event of caspase activation. When stably or transiently overexpressed, ATF3 markedly accelerated the drug-induced apoptosis and enhanced caspase protease activation. ATF3 strongly downregulated CRE-dependent transcription, while ATF3 did not affect the expression levels of Bcl-2, Bcl-x, or Bax. Our present results indicate that ATF3 plays a critical role in accelerating caspase protease activation and apoptosis. Since CRE-dependent transcription functions as cell survival signaling, ATF3 could control the upstream signaling of apoptosis by repressing CRE-dependent gene expression of cell survival factors.

Antifungal substances against pathogenic fungi, talaroconvolutins, from talaromyces convolutus.

The dichloromethane extract of Talaromyces convolutus cultivated on barley exhibited antifungal activity against Candida albicans. In the course of a search for the active compounds, four new tetramic acid derivatives, talaroconvolutins A (1), B (2), C (3), and D (4), were isolated along with ZG-1494alpha (5), and mitorubrin derivatives. The structures of talaroconvolutins A-D (1-4) were established on the basis of spectroscopic and chemical investigations and chemical correlations. The antifungal activity of the talaroconvolutins against the pathogenic fungi Aspergillus fumigates, Aspergillus niger, C. albicans, and Cryptococcus neoformans was determined.

Subcutaneous phaeohyphomycosis caused by Phaeoacremonium rubrigenum in an immunosuppressed patient.

BACKGROUND: Phaeohyphomycosis refers to infection by dematiaceous fungi with pigmented hyphae or yeast-like cells in the tissue. In humans, this disease is usually considered to be an opportunistic infection. The causal agents of phaeohyphomycosis include numerous species belonging to different genera and they are increasing as a result of the development of intensive medical therapy. OBSERVATION: We report the case of a 61-year-old Japanese female under corticosteroid treatment for malignant rheumatoid arthritis. An asymptomatic subcutaneous tumor developed on the back of her left foot. Histological examination of the excised material revealed mixed cell granuloma (H&E) and the presence of branched hyphal elements (periodic acid-Schiff). A fungus grown in pure culture was identified as Phaeoacremonium rubrigenum. CONCLUSION: The hyphomycete genus, Phaeoacremonium, was proposed in 1996 by Crous et al. Three species belonging to this genus have been isolated from clinical specimens: P. inflatipes, from a human toenail, human synovial fluid and human mycetoma of the foot, P. parasiticum, from a subcutaneous lesion on a kidney transplant patient and several other sources, and P. rubrigenum, from a human patient with pneumonia. To our knowledge, however, this is the first report of phaeohyphomycosis caused by Phaeoacremonium rubrigenum.

Subcutaneous phaeohyphomycosis caused by Geniculosporium species; a new fungal pathogen.

A 70-year-old Japanese timberworker dealing with imported timber from the U.S.A. and Russia had an asymptomatic subcutaneous nodule with a small fistula on his left knee. Histopathological examination of the nodule revealed brownish hyphal elements in encapsulated pyogranuloma. The fungus isolated from a discharge of the fistula and an excised specimen of the lesion was identified as Geniculosporium sp., which represents a conidial state (anamorph) of several genera such as Anthostomella, Biscogniauxia, Euepixylon, Leprieuria, Nemania, Phylacia and Rosellina in the Xylariaceae. Whereas this dematiaceous hyphomycete is commonly found on decaying wood and bark of various trees, to our knowledge, this is the first case of a phaeomycotic cyst caused by fungi belonging to the genus Geniculosporium.

[A case of cutaneous pseudallescheriosis resembling sporotrichosis].

An 83-year-old man with aplastic anemia and steroid induced diabetes mellitus developed multiple nodules with pus on the back of his right hand and forearm. He had been treated with a daily dose of 30mg prednisolone for 2 months. These lesions had appeared a month before his visit. The histopathological findings revealed dermal abscesses containing hyphal structures, lymphocytes, histiocytes and giant cells. Grocott-Methenamine stain demonstrated abundant fungal elements. In culture, colonies grow rapidly and produce a white, cottony mycelium which later becomes gray in color. Microscopically, ovoid and pyriform conidia are produced at the ends of long slender conidiophores. On PCA agar, synnemata with small conidal heads developed at 37 degrees C but cleistothecia did not appear. The patient did not respond to itraconazole therapy, however, with hyperthermic treatment, the eruptions gradually healed. Based on these findings, this fungal infection was diagnosed as pseudallescheriosis.

Isolation of Rhizopus microsporus var. rhizopodiformis in the ulcer of human gastric carcinoma.

Gastric zygomycosis in a patient with gastric cancer was described. Colonization was diagnosed because broad sparsely septate hyphae were identified in the necrotic tissue of gastric biopsied specimens. No fungus was present within viable tissue. Positive cultures for Rhizopus microsporus var. rhizopodiformis were obtained from fragments of biopsied specimens. Members of the Mucorales have rarely been isolated from the culture of endoscopically biopsied materials.

Isolation and structures of antibacterial binaphtho-alpha-pyrones, talaroderxines A and B, from Talaromyces derxii.

New compounds designated talaroderxines A (1) and B (2) were isolated from a new heterothallic ascomycetous fungus, Talaromyces derxii, cultivated on rice. The structures of 1a and 1b were elucidated by means of spectroscopic examination and chemical reactions. Talaroderxines A (1a) and B (1b) are atropisomers of a 6,6'-binaphtho-alpha-pyrone derivative, and have strong antibacterial activity against Bacillus subtilis.

Experimental acute poisoning in mice induced by emestrin, a new mycotoxin isolated from Emericella species.

The effects of emestrin (EMS), a secondary metabolite of the Emericella species, on male ICR mice were examined. The intraperitoneal LD50 values of EMS were 17.7 and 13.0 mg/kg at 24 and 48 hr, respectively. The target organs of EMS were the heart, liver and thymus. In doses over 30 mg/kg the experimental animals died from cardiac failure shortly after the injections. Several survivors that were given EMS in doses under 20 mg/kg showed severe centrilobular necrosis in the liver at 24 hr. Marked degeneration of mitochondria was seen in electron micrographs of both cardiac muscle cells and hepatocytes. In the degenerated hepatocytes, prominent proliferation of RER, membrane-limited inclusions containing both ribosome-like granules and RER, and fenestrated lamella-like structures were observed. Massive necrosis of lymphocytes was always observed in the cortical layer of the thymus of the survivors within 24 hr, while bilateral adrenalectomized mice showed no discernible pathomorphological changes in the lymphoid tissues. Pretreatment of mice with diethyl maleate increased the incidence and severity of hepatic necrosis, whereas that with either cysteine or CoCl2 reduced the severity of centrilobular necrosis of the liver. Pretreatment with phenobarbital had no significant effect on EMS-induced hepatic lesions.

Pulmonary aspergillosis due to Aspergillus terreus combined with staphylococcal pneumonia and hepatic candidiasis.

A female patient with systemic lupus erythematosus (SLE) developed pulmonary aspergillosis with staphylococcal pneumonia and hepatic candidiasis. Aspergillus terreus, which is a rare causative organism of pulmonary aspergillosis, was identified from a pulmonary lesion by culture. The aleurioconidium production, a characteristic of the genus Aspergillus sect. terrei, was demonstrated on short and irregular hyphal features in tissue sections. This report is the first of a combined case of pulmonary aspergillosis due to A. terreus with infections caused by other microorganisms.

Paecilotoxin production in clinical or terrestrial isolates of Paecilomyces lilacinus strains.

The production of paecilotoxin from various isolates of Paecilomyces lilacinus was studied using three different media and high performance liquid chromatography (HPLC). Alkaline medium was found to be suitable for the production of the toxins. Among 20 strains tested, 19 including four clinical isolates were found to produce the toxins. Production patterns of paecilotoxins were very similar in each strain and the main toxins were A and B.

Isolation of a new tremorgenic indoloditerpene, 1'-O-acetylpaxilline, from Emericella striata and distribution of paxilline in Emericella spp.

The distribution of a tremorgenic mycotoxin, paxilline (1), was investigated in 19 species belonging to the genus Emericella. It was found that Emericella desertorum, E. foveolata, and E. striata produced paxilline (1). A new type of indoloditerpene, emindole DA (4), was also found in E. quadrilineata. A new tremorgenic indoloditerpene, 1'-O-acetylpaxilline (3), was isolated from the mycelium of E. striata. Its structure was established on the basis of spectroscopic investigations.

Effect of roasting on ochratoxin A level in green coffee beans inoculated with Aspergillus ochraceus.

The heat stability of ochratoxin A in green coffee beans inoculated with Aspergillus ochraceus was studied. Heat treatment (roasting) at 200 degrees C for 10 or 20 min reduced the levels of ochratoxin A by only 0-12% in the dried whole beans. Almost all of the ochratoxin A was infused into the coffee decoction when the roasted samples were ground and extracted with boiling water. Therefore, the reduction of ochratoxin A concentration of contaminated coffee beans by roasting under these conditions is ineffective.

Subcutaneous abscesses caused by Ochroconis gallopavum.

A subcutaneous fungus infection that occurred in a patient with an acute myeloblastic leukemia was found to be caused by Ochroconis gallopavum (Dactylaria gallopava). In histological sections dematiaceous, septate hyphae were present. A darkly pigmented fungus was isolated on Sabouraud glucose medium. The mycological features of the causative agent were typical of O. gallopavum. The patient died after 6 months of treatment with antileukemic drugs and 5-fluorocytosine. At autopsy, tissue sections revealed leukemic infiltrates in most of the internal organs but fungal invasion was not detected.