Seed production and storage for endangered Morus boninensis using an ex-situ living collection.

Meaningful conservation techniques for the endangered Morus boninensis require seed production and storage, since in situ purebred seed production is infrequent in Ogasawara Islands (World Natural Heritage Site) in Japan. Ex situ living M. boninensis specimens produced seeds by open- and artificial-pollination in a greenhouse. Seed desiccation tolerance and storage characteristics were examined. Open-pollinated seeds were dried at 75, 33 and 8% relative humidity (RH) for 5 days then cryopreserved at -170 °C. Artificially pollinated seeds were dried at 8% RH then stored cryogenically at -170 °C and in freezers at -80 or -20 °C for 6 months. Germination rates were compared among seeds before and after drying and storage. Germination rates of undried seeds were 83% for open-pollinated seeds and 65% for artificially pollinated seeds. M. boninensis seeds exhibited the characteristic high desiccation tolerance of orthodox seeds and maintained a high germination rate after drying to a moisture content of approximately 4%. Cryopreservation of open-pollinated dry seeds with moisture contents of ca. 12, 7 and 4% maintained germination rates, while cryopreservation of undried seeds (moisture content ca. 44%) resulted in no germinability. Cryogenic and freezer storage of artificially pollinated dry seeds for 6 months resulted in maintenance of germination rates comparable to undried seeds before storage. Seed production was enabled by ex situ M. boninensis collection. The present study also developed a method for low temperature seed storage. Ex situ living tree collection represents a feasible seed resource for reintroduction and restoration programmes for M. boninensis.

Sodium-glucose cotransporter 2 inhibitor luseogliflozin improves glycaemic control, assessed by continuous glucose monitoring, even on a low-carbohydrate diet.

This randomized, double-blind, placebo-controlled, crossover study was the first to determine the effects of luseogliflozin in combination with a low-carbohydrate diet (LCD) on 24-h glucose variability, assessed by continuous glucose monitoring (CGM). A total of 18 Japanese patients with type 2 diabetes were randomized into two groups, in which patients first received luseogliflozin 2.5 mg once daily then placebo for 8 days each, or vice versa. Patients took luseogliflozin or placebo with a normal-carbohydrate diet (NCD) on day 7 and with the LCD on day 8. CGM was performed on both days. Luseogliflozin significantly reduced glucose exposure in terms of the area under the curve over the course of 24 h when administered with the NCD (difference vs placebo: -555.6 mg/dl·h [1 mg/dl = 0.0556 mmol/l]; p < 0.001) or with the LCD (-660.7 mg/dl·h; p < 0.001). No hypoglycaemia was observed over 24 h with either diet. Although glucose levels were lower with the LCD than with the NCD in the placebo treatment period, luseogliflozin with the LCD improved glycaemic control throughout the day to nearly the same extent as luseogliflozin with the NCD, without inducing hypoglycaemia.

Effects of luseogliflozin, a sodium-glucose co-transporter 2 inhibitor, on 24-h glucose variability assessed by continuous glucose monitoring in Japanese patients with type 2 diabetes mellitus: a randomized, double-blind, placebo-controlled, crossover study.

The aim of the present study was to determine the effects of luseogliflozin on 24-h glucose levels, assessed by continuous glucose monitoring, and on pharmacodynamic variables measured throughout the day. In this double-blind, placebo-controlled, crossover study, 37 patients with type 2 diabetes mellitus inadequately controlled with diet and exercise were randomized into two groups. Patients in each group first received luseogliflozin then placebo for 7 days each, or vice versa. After 7 days of treatment, the mean 24-h glucose level was significantly lower with luseogliflozin than with placebo [mean (95% confidence interval) 145.9 (134.4-157.5) mg/dl vs 168.5 (156.9-180.0) mg/dl; p < 0.001]. The proportion of time spent with glucose levels ≥70 to ≤180 mg/dl was significantly greater with luseogliflozin than with placebo [median (interquartile range) 83.2 (67.7-96.5)% vs 71.9 (46.9-83.3)%; p < 0.001] without inducing hypoglycaemia. The decrease in glucose levels was accompanied by reductions in serum insulin levels throughout the day.

Smoking-related changes in the background lung of specimens resected for lung cancer: a semiquantitative study with correlation to postoperative course.

AIMS: To assess the pathological findings in lobectomy specimens, to correlate them with smoking history and postoperative course and to compare the findings with those in smoking-related interstitial lung disease. METHODS AND RESULTS: Patients who had undergone lobectomy for lung cancer were reviewed. Subjects included 230 non-smokers and 587 smokers, of whom 572 had a known smoking index (SI). They were classified into mild, moderate and heavy smokers. Centrilobular emphysema (CLE), respiratory bronchiolitis, airspace enlargement with fibrosis (AEF), the presence of foci resembling usual interstitial pneumonia pattern (UIP/P) and the rate of postoperative respiratory failure were assessed. The incidence of AEF was 6.5% in mild smokers, and 17.7% in moderate smokers (P < 0.01) with lower lobe predominance. There were significant correlations (P < 0.01) between AEF and CLE and AEF and UIP/P. The rate of respiratory failure after lobectomy was 6%, and 10% in patients having UIP/P with or without AEF, but was not seen in patients with AEF alone (P < 0.01). CONCLUSIONS: AEF is an important smoking-related change in the lung that appears to correlate with the smoking history, and its distinction from UIP/P may be important.

[Invasive pulmonary aspergillosis following influenza A infection].

A 55-year-old man with arc welder's pneumoconiosis who presented with bilateral pneumonia was admitted to our hospital. Streptococcus pneumoniae and Aspergillus fumigatus were cultured from sputum on admission. The patient was treated with antibiotics and an anti-fungal agent, but his chest radiograph shadows became exacerbated, and he died of respiratory failure on the fourth day of hospitalization. Histological examination of postmortem lung tissue revealed necrotizing aspergillous pneumonia. The results of hemagglutination inhibition tests for influenza A (H3 N2) were x16 in September 1999, and x512 on the third day of hospitalization. We diagnosed this patient's condition as invasive pulmonary aspergillosis associated with influenza A viral infection. The suppression of cellular immunity, lymphocytopenia, and destruction of airways-mucociliary transport induced by influenza A viral infection were suspected to have predisposed him to aspergillus superinfection.

[A case of pulmonary lymphangioleiomyomatosis in an elderly woman].

We encountered a postmenopausal woman in whom pulmonary lymphangioleiomyomatosis was pathologically diagnosed. Cases of pulmonary lymphangioleiomyomatosis are rare in postmenopausal women. The patient was the oldest in whom such a case was reported in the literature. Pulmonary lymphangioleiomyomatosis should be on the list of differential diagnoses for cases of pulmonary cystic lesions, even in postmenopausal women.

[Pathologic findings both before and after steroid therapy in a case of desquamative interstitial pneumonia].

A 62-year-old man was admitted to our hospital because of dyspnea on effort. Histological findings from video-assisted thoracoscopic surgical biopsy were desquamative interstitial pneumonia (DIP). Two months after the introduction of steroid therapy, the patient underwent right middle lobe lobectomy for primary lung cancer. The histologic findings in this lobe revealed non-specific interstitial pneumonia, not characteristic of DIP. This case was the first in which histologic examination revealed DIP both before and after steroid therapy.

Tautomycetin is a novel and specific inhibitor of serine/threonine protein phosphatase type 1, PP1.

Here we isolated tautomycetin, TC, and examined its phosphatase inhibitory activity. Recently we have reported that the left-hand moiety of tautomycin, TM, and the right one containing the spiroketal are essentially required for inhibition of protein phosphatase, PP, and induction of apoptosis, respectively. TC is structurally almost identical to TM except that TC is lacking the spiroketal, which has the potential apoptosis-inducing activity. TC specifically inhibited PP1 activity, IC50 values for purified PP1 and PP2A enzymes being 1.6 and 62 nM, respectively, whereas the IC50 values of TM were 0.21 and 0.94 nM, respectively. These results demonstrate that TC is the most specific PP1 inhibitor out of over 40 species of natural phosphatase inhibitors reported, strongly suggesting that TC is a novel powerful tool to elucidate the physiological roles of PP1 in various biological events.

Stilbenoids in lianas of Gnetum parvifolium.

Five new stilbene dimers were isolated from the lianas of Gnetum parvifolium in addition to known stilbenoids. The structures of the compounds were established on the basis of spectroscopic evidence, including long-range coupling and nuclear Overhauser effect experiments, in NMR spectrum. Among the isolates, 2b-hydroxyampelopsin F showed potent inhibitory activity in the Maillard reaction.

[A case of drug-induced interstitial pneumonitis in rheumatoid arthritis treated with bucillamine].

We report a case of bucillamine-induced interstitial pneumonitis in a 57-year-old woman. Rheumatoid arthritis was diagnosed in May 1999, and she was treated with bucillamine from June 1999, with a favorable outcome. After complaining of cough, fever, and dyspnea in October, she was admitted to this hospital. Blood gas analysis showed severe hypoxemia. The chest CT revealed both bilateral diffuse ground-glass opacity along the bronchovascular bundles, and thickening of the interlobular septa. We suspected bucillamine-induced interstitial pneumonitis from the findings of the CT scan, BALF and TBLB, and also from the improvement of PaO2 after the withdrawal of bucillamine. We treated the patient with prednisolone, and a favorable response was noted. A lymphocyte stimulation test using bucillamine was positive. A video-assisted thoracic surgery lung biopsy showed findings compatible with acute interstitial pneumonia without the association of hyaline membrane formation. A focal fibrosis was also observed. We believe that this is the only reported case of pathologically proven bucillamine-induced interstitial pneumonitis, in which a surgical lung biopsy was performed.

Pulmonary nocardiosis during immunosuppressive therapy for idiopathic pulmonary fibrosis.

Nocardiosis is a subacute or chronic suppurative infection caused by Nocardia species. Although it is more common in immunocompromised hosts, idiopathic pulmonary fibrosis (IPF) has not been recognized as a predisposing factor for nocardial infection. We report a case of IPF, in which pulmonary nocardiosis developed during treatment with prednisolone and cyclophosphamide. The risk of pulmonary nocardiosis may be increased in cases of IPF on immunosuppressive therapy. Since IPF often accompanies lung carcinoma, it is important to correctly differentiate nocardiosis from carcinoma.

[Familial summer-type hypersensitivity pneumonitis in a grandfather and his two-and-a-half-year-old grandson].

We encountered a family in which two of the six members, the grandfather and his grandson, had summer-type hypersensitivity pneumonitis in late summer. Chest computed tomography of these patients showed small, diffuse nodular shadows in both lung fields, with mosaic-like opacities in the grandfather and ground glass opacity of both lung fields in the grandson. Bronchoalveolar lavage fluid from the grandfather disclosed high total cell counts, high percentages of lymphocytes, and a low ratio of CD 4+ cells to CD 8+ cells. Transbronchial lung biopsy specimens obtained from the same patient revealed alveolitis with non-caseous epithelioid cell granulomas in the interstitium and Masson bodies in the alveolar septa. These two patients recovered spontaneously after hospital admission. They had positive results in provocation tests for their home and were positive for serum anti-Trichosporon cutaneum (T. asahii, T. mucoides) antibodies. Both cases were accordingly diagnosed as summer-type hypersensitivity pneumonitis.

New types of liposidomycins that inhibit bacterial peptidoglycan synthesis and are produced by Streptomyces. II. Isolation and structure elucidation.

Various new liposidomycins were isolated from a culture of the strain Streptomyces sp. SN-1061M by changing medium components and they were classified into four types (I-IV) based on their structures. They were purified by butanol extraction, silica gel and LH-20 column chromatographies, and high performance liquid chromatography on ODS columns. Type (I) has the original structure which has sulfate and 3-methylglutaric acid moieties. Type (II) has no 3-methylglutaric acid moiety and type (III) has no sulfate moiety. Type (IV) has neither moiety. Type (III) and (IV) compounds, which have no sulfate moiety, exhibited more potent antimicrobial activity.

Different moieties of tautomycin involved in protein phosphatase inhibition and induction of apoptosis.

The effects of tautomycin and its derivatives on protein phosphatases PP1 and PP2A and their apoptosis-inducing activity toward human leukemia Jurkat cells were examined, and the relationship between chemical structure and function was discussed. Among the compounds we examined, tautomycin was the most potent inhibitor and the most effective inducer of apoptosis. It inhibited PP1 and PP2A enzymatic activity concentration-dependently with IC50 values of 20 and 75 pM, respectively, in the presence of 0.01% Brij-35, and an LC50 value of 1 microM. Esterification of the anhydride moiety of tautomycin markedly increased the IC50 for the protein phosphatases. The C1'-C7' fragment of tautomycin had no inhibitory effect, but the fragment containing the C22-C26 moiety was inhibitory. These results suggest that the C22-C26 moiety is essential for inhibition of protein phosphatase activity and that the anhydride moiety enhances the inhibition. However, the esterification of the anhydride did not decrease, nor did the inclusion of the C22-C26 moiety increase the apoptosis-inducing activity. On the other hand, the C1-C18 moiety of tautomycin was essential for induction of apoptosis, and the conformation and the arrangement of functionalities of the C18-C26 carbon chain affected the apoptosis activity. However, modification of C1-C18, C1-C21, or C1-C26 compounds had little effect on phosphatase inhibitory activity. Our results strongly suggest that different moieties of tautomycin are involved in protein phosphatase inhibition and induction of apoptosis.

Synthesis and morphological reversion activity on srctsNRK cells of pyrimidinylpropanamide antibiotics, sparsomycin, sparoxomycin A1, A2, and their analogues.

Three pyrimidinylpropanamide antibiotics sparsomycin (1), sparoxomycins A1, A2 (2, 3), and also six analogues (4-9) have been synthesized by employing asymmetric sulfide oxidation conditions as a key step. Sparsomycin (1) and its alkyl analogues (5-7) showed higher morphological reversion activities on srctsNRK cells than 2 and 3.

Facile Synthesis of Cyanogen Glycosides (R)-Prunasin, Linamarin and (S)-Heterodendrin.

A facile synthetic route is described to cyanogenic glycosides (R)-prunasin, linamarin and (S)-heterodendrin from O-(2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl)trichloroace- timidate and the corresponding α-hydroxyamides by a 3-step reaction of glycosylation, cyanohydrin formation by dehydration of carboxamides, and deprotection.

[Clinical and diagnostic characteristics of Legionella pneumonia].

We have characterized clinical and diagnostic features in 18 cases of Legionella pneumonia. Age average of patients was 62.0 years old (male:female = 14:4) and underlying diseases were observed in 12 patients. Legionella pneumonia were diagnosed in 3, 5, 8 and 9 cases by culture, serum antibody measurement, urinary antigen detection and PCR, respectively. Sixteen cases were caused by L. pneumophila, while the other 2 cases were L. bozemanii pneumonia and L. pneumophila or L. dumoffii pneumonia. Chest X-rays of those patients showed multiple pneumonia shadows in 14 cases, alveolar shadows in 10 cases, pleural effusion in 5 cases. Blood-gas analysis on admission indicated hypoxemia in all cases with abnormal A-a DO2. Laboratory findings showed abnormal data in WBC, CRP, LDH, CPK and liver function tests (ex. GOT, GPT) in most cases. Serum antibody testing showed positive by 5 weeks after onset of pneumonia, but 10 cases of Legionella pneumonia diagnosed by other techniques were judged to be negative. In urinary antigen detection test, 6 and 2 cases showed positive 1 and 4 weeks after onset of pneumonia, respectively. Macrolide antibiotics were administered in all cases during the episode, but delay of macrolide administration was observed in 3 of 4 cases of dead outcome. Serum antibody measurement, urinary antigen detection and PCR, in addition to culture to bacteria, may be required for exact diagnosis of Legionella infection.

Effectiveness of Tc-99m MIBI scintigraphy in diagnosing lung cancer.

The use of Tc-99m hexakis 2-methoxy isobutyl isonitrile (MIBI) scintigraphy for qualitative diagnosis was examined. A total of 37 lesions from 31 patients with respiratory disease (19 lesions from 19 primary lung cancer patients, 8 lesions from 3 metastatic lung cancer patients and 10 lesions from benign lung disease patients) were examined. Positive results were seen in 89.4% of the primary lung cancer lesions, 50.0% of the metastatic lung cancer lesions, and 20.0% of the benign lung disease lesions. The positive rate of 77.7% for malignant lung cancer was significantly higher (P < 0.01) than that for benign lung cancer. In particular, for lesions with a tumor diameter of 1.0 cm or more, the positive rate for malignant lung cancer lesions (95.2%) was significantly higher (P < 0.001) than that for benign lung disease lesions (25.0%). These results thus indicate Tc-99m MIBI scintigraphy to be useful for qualitative diagnosis of lung peripheral coin lesions with a diameter of 1.0 cm or more.