ABSTRACT
AIMS: The present study was conducted to screen for psychrophilic yeasts that are able to degrade pectin compounds at low temperature, and to examine the cold-active pectinolytic enzymes produced by the isolated psychrophilic yeasts. METHODS AND RESULTS: Psychrophilic yeasts, which grow on pectin as a sole carbon source, pectinolytic-psychrophilic yeast (PPY) strains PPY-3, 4, 5 and 6, were isolated from soil from Abashiri (Hokkaido, Japan). The sequences of 28S rDNA D1/D2 of strains PPY-3 and 4 indicated a taxonomic affiliation to Cryptococcus cylindricus and Mrakia frigida, respectively, strains PPY-5 and 6 belonged to Cystofilobasidium capitatum. The isolated strains were able to grow on pectin at below 5 degrees C, and showed the activities of several cold-active pectinolytic enzymes. CONCLUSION: The findings of this study indicate the possibility that the isolated strains produce novel pectinolytic enzymes that are able to degrade pectin compounds at low temperature. SIGNIFICANCE AND IMPACT OF THE STUDY: It is possible that the cold-active pectinolytic enzymes from the isolated strains can be applied to the food industry, e.g. the clarification of fruit juice below 5 degrees C.
Subject(s)
Basidiomycota , Cold Temperature , Cryptococcus , Pectins/metabolism , Soil Microbiology , Basidiomycota/classification , Basidiomycota/enzymology , Basidiomycota/genetics , Basidiomycota/isolation & purification , Carboxylic Ester Hydrolases/metabolism , Cryptococcus/classification , Cryptococcus/enzymology , Cryptococcus/genetics , Cryptococcus/isolation & purification , DNA, Ribosomal/analysis , Enzyme Stability , Glycoside Hydrolases/metabolism , Molecular Sequence Data , Phylogeny , Polysaccharide-Lyases/metabolism , RNA, Ribosomal, 28S/genetics , Sequence Analysis, DNA , Substrate SpecificityABSTRACT
BACKGROUND: The effectiveness of losartan for the treatment of leucocyte entrapment in the retinal microcirculation of diabetic rats was evaluated quantitatively. METHODS: After diabetes was induced by injection of streptozotocin (STZ), the rats were divided into two subgroups. The first subgroup (n = 6), received no medications; the second subgroup (n = 6) was given fresh drinking water supplemented with losartan (5 mg/kg/day) for 4 weeks. Six rats that were not injected with STZ or given medications served as controls. 4 weeks after intervention, leucocyte dynamics in the retina were observed using acridine orange digital fluorography. Leucocyte entrapment in the retina was compared among the three groups. RESULTS: In the untreated diabetic rats, the number of trapped leucocytes (6.1 (SD 1.4) cells/mm(2)) increased significantly compared with control rats (2.8 (1.2) cells/mm(2); p = 0.005) and diabetic rats treated with losartan (3.1 (0.9) cells/mm(2); p = 0.0002). CONCLUSIONS: Losartan, an AT1 angiotensin II receptor antagonist, inhibited increased leucocyte entrapment in the diabetic retina. The authors demonstrated that losartan may have therapeutic efficacy in preventing development of diabetic retinopathy. Further clinical studies of the effect of the angiotensin receptor antagonist on preventing development of diabetic retinopathy are needed.
Subject(s)
Angiotensin Receptor Antagonists , Diabetes Mellitus, Experimental/physiopathology , Leukocytes/drug effects , Losartan/pharmacology , Retinal Vessels/drug effects , Acridine Orange , Animals , Fluorescence , Male , Microcirculation/drug effects , Rats , Rats, Inbred BNABSTRACT
A new dimeric aporphine alkaloid, phoenicanthusine (1), and six known alkaloids were isolated from the stem bark of Phoenicanthus obliqua. The structure of 1 was elucidated by 1D ((1)H, (13)C) and 2D ((1)H-(1)H COSY, HMQC, HMBC, and NOESY) NMR and HRMS studies. Phoenicanthusine represents the first example of a N-6--C-4' and C-7--C-5' linked dimeric aporphine alkaloid.
Subject(s)
Annonaceae/chemistry , Aporphines/isolation & purification , Plants, Medicinal/chemistry , Aporphines/chemistry , Chromatography, Thin Layer , Molecular Conformation , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Plant Stems/chemistry , Spectrophotometry, Infrared , Spectrophotometry, Ultraviolet , Sri LankaABSTRACT
Five known sesquiterpenoids, solavetivone, lubimin, lubiminoic acid, aethione and lubiminol were isolated from the root exudates recovered from Solanum aethiopicum by a newly proposed method using charcoal. Quantitative analysis of the sesquiterpenoids in the root exudates of S. aethiopicum and S. melongena suggested that relatively large amounts of the sesquiterpenoids were exuded from the roots. Antifungal activity of the sesquiterpenoids against Fusarium oxysporum and Verticillium dahliae was also examined.
Subject(s)
Antifungal Agents/chemistry , Antineoplastic Agents, Phytogenic/chemistry , Plant Extracts/chemistry , Plant Roots/chemistry , Sesquiterpenes/chemistry , Solanaceae/chemistry , Antifungal Agents/isolation & purification , Antineoplastic Agents, Phytogenic/isolation & purification , Fungi/drug effects , Furans/chemistry , Furans/isolation & purification , Furans/pharmacology , Gas Chromatography-Mass Spectrometry , Magnetic Resonance Spectroscopy , Microbial Sensitivity Tests , Molecular Conformation , Plant Extracts/isolation & purification , Sesquiterpenes/isolation & purification , Sesquiterpenes/pharmacology , Species SpecificityABSTRACT
Sixty-eight water and methanol extracts from 34 Chinese herbal drugs, most of which are used for inflammatory diseases, were screened for their inhibitory effects on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated J774.1 macrophages and in LPS/interferon (IFN)-gamma-stimulated mouse peritoneal exudate macrophages. Among the extracts, methanol extracts of Myristica fragrans, Plantago asiatica, Rubia cordifolia, and Zanthoxylum bungeanum showed significant inhibition in J774.1 macrophages, while in mouse peritoneal exudate macrophages, water extracts of Ru. cordifolia and Scutellaria baicalensis and methanol extracts of Angelica megaphylla, My. fragrans, and Z. bungeanum inhibited the NO production. Among them, inhibition of water extract of Sc. baicalensis was found to be mainly due to direct scavenging of NO radicals, through an examination of its scavenging activity on PAPA NONOate-generated NO radicals, while water extract of Ru. cordifolia and methanol extracts of An. megaphylla, My. fragrans, P. asiatica, and Z. bungeanum showed inhibition on iNOS mRNA expression. At last, an inhibitory compound on iNOS mRNA expression was isolated from a methanol extract of Z. bungeanum and identified as 4-O-beta-D-glucopyranosyldihydroferulic acid by NMR spectral analyses and chemical synthesis.
Subject(s)
Drug Evaluation, Preclinical/methods , Drugs, Chinese Herbal/pharmacology , Macrophages, Peritoneal/drug effects , Nitric Oxide Synthase/biosynthesis , Nitric Oxide/biosynthesis , Animals , Cell Survival , Cells, Cultured , Drugs, Chinese Herbal/isolation & purification , Macrophages, Peritoneal/metabolism , Male , Mice , Mice, Inbred BALB C , Nitric Oxide Synthase Type II , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
13 quinic acid derivatives along with caffeic acid, methyl caffeate, myo-inositol, bis[5-formylfurfuryl] ether and 6,7-dihydroxycoumarin were isolated from the ethanol extract of the flower buds of Lonicera bournei Hemsl., among which 8 compounds were firstly obtained from this genus. The effects of different solvent soluble fractions of the ethanol extract and the pure compounds on heptocyte death induced by D-galactosamine (D-GalN)/tumor necrosis factor alpha (TNF-alpha) were studied, and the structure-activity relationships were also discussed.
Subject(s)
Caffeic Acids/isolation & purification , Cells, Cultured/drug effects , Ethers/isolation & purification , Glycosides/isolation & purification , Hepatocytes/drug effects , Liver/cytology , Liver/drug effects , Plants, Medicinal/chemistry , Quinic Acid/analogs & derivatives , Quinic Acid/isolation & purification , Umbelliferones/isolation & purification , Animals , Apoptosis , Caffeic Acids/chemistry , Caffeic Acids/pharmacology , Drugs, Chinese Herbal , Ethers/pharmacology , Galactosamine/pharmacology , Glycosides/chemistry , Glycosides/pharmacology , In Vitro Techniques , Inositol/pharmacology , Magnetic Resonance Spectroscopy , Male , Medicine, Chinese Traditional , Mice , Mice, Inbred Strains , Molecular Structure , Quinic Acid/chemistry , Quinic Acid/pharmacology , Spectrophotometry, Infrared , Structure-Activity Relationship , Tumor Necrosis Factor-alpha/pharmacology , Umbelliferones/chemistry , Umbelliferones/pharmacologyABSTRACT
The methanol extract of Vietnamese ginseng (Panax vietnamensis) was found to possess hepatocytoprotective effects on D-galactosamine (D-GalN)/tumor necrosis factor-alpha (TNF-alpha)-induced cell death in primary cultured mouse hepatocytes. Further chemical investigation of the extract afforded two new dammarane-type triterpene saponins, ginsenoside Rh(5) (1) and vina-ginsenoside R(25) (2), as well as eight known dammarane-type triterpene saponins, majonoside R(2) (3), pseudo-ginsenoside RT(4) (4), vina-ginsenosides R(1) (5), R(2) (6), and R(10) (7), ginsenosides Rg(1) (8), Rh(1) (9), and Rh(4) (10), and a known sapogenin protopanaxatriol oxide II (11). Their structures were elucidated on the basis of spectral analysis. In addition, by the using LC-electrospray ionization (ESI)-MS method, five known saponins, ginsenosides Rb(1), Rb(2), Rc, Rd, and Re (12--16), were also identified in the extract. Among the compounds isolated, majonoside R(2) (3), the main saponin in Vietnamese ginseng, showed strong protective activity against D-GalN/TNF-alpha-induced cell death in primary cultured mouse hepatocytes. This demonstrates that the hepatocytoprotective effect of Vietnamese ginseng is due to dammarane-type triterpene saponins that have an ocotillol-type side chain, a characteristic constituent of Vietnamese ginseng.
Subject(s)
Liver/drug effects , Panax/chemistry , Plants, Medicinal , Saponins/isolation & purification , Triterpenes/isolation & purification , Animals , Cell Death/drug effects , Chromatography, High Pressure Liquid , Liver/cytology , Mice , Molecular Structure , Saponins/chemistry , Saponins/pharmacology , Spectrometry, Mass, Electrospray Ionization , Triterpenes/chemistry , Triterpenes/pharmacology , Tumor Necrosis Factor-alpha/pharmacologyABSTRACT
To elucidate the antibacterial activity of Gosyuyu, the crude extract from the fruit of Evodia rutaecarpa, a Chinese herbal medicine, has been fractionated chromatographically, and each fraction was assayed for antibacterial activity against Helicobacterpylori (H. pylori) in vitro. As the result, a single spot having marked antibacterial activity against H. pylori was obtained and the chemical structure was analyzed. The isolated compound was revealed to be a novel alkyl quinolone alkaloid based on the solubility, IR spectra, NMR analysis and mass spectrometric data after purification by TLC of silica. We compared the antimicrobial activity of this compound with that of other antimicrobial agents and examined susceptibility of various intestinal pathogens. As the result, the new quinolone compounds obtained from Gosyuyu extracts were found to be a mixture of two quinolone alkaloids, 1-methyl-2-[(Z)-8-tridecenyl]-4-(1H)-quinolone and 1-methyl-2-[(Z)-7-tridecenyl]-4-(1H)-quinolone (MW: 339), reported previously. The minimum inhibitory concentration (MIC) of these compounds against reference strains and clinically isolated H. pylori strains were less than 0.05 microg/ml, which was similar to the MIC of amoxicillin and clarithromycin that are used worldwide for the eradication of H. pylori, clinically. Furthermore, it was noted that the antimicrobial activity of these compounds was highly selective against H. pylori and almost non-active against other intestinal pathogens. The above results showed that these alkyl methyl quinolone (AM quinolones) alkaloids were useful for the eradication of H. pylori without affecting other intestinal flora.
Subject(s)
Alkaloids/pharmacology , Anti-Infective Agents/pharmacology , Drugs, Chinese Herbal/pharmacology , Helicobacter pylori/drug effects , Quinolines/pharmacology , Quinolones/pharmacology , Alkaloids/chemistry , Alkaloids/isolation & purification , Anti-Infective Agents/chemistry , Anti-Infective Agents/isolation & purification , Chemical Fractionation , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/isolation & purification , Evodia , Fruit/chemistry , Helicobacter pylori/isolation & purification , Humans , Microbial Sensitivity Tests , Molecular Structure , Plant Extracts , Quinolines/chemistry , Quinolines/isolation & purification , Quinolones/chemistry , Quinolones/isolation & purification , Solvents , Stomach Ulcer/microbiologyABSTRACT
An overoxidized polypyrrole colloid, which can recognise enantiomers of amino acids has been prepared by a newly developed molecular imprinting technique. A polypyrrole colloid, which had been polymerised from a mixture of pyrrole (monomer), polyvinylpyrrolidone (steric stabiliser), peroxodisulfate (oxidant) and L-lactate (dopant), was overoxidized to create a dopant-complementary cavity. The enantioselectivity of the overoxidized colloid was evaluated by comparing the uptake of L-alanine and L-cysteine with that of the respective D-enantiomers. The L/D uptake ratios for these amino acids were about 2, while phenylalanine showed suppressed uptake for both the enantiomers. The absence of phenylalanine uptake can be explained in terms of the molecular size, which is too large to be accommodated by the cavity created by L-lactate. In contrast, a colloid templated with L-phenyllactate took up L-phenylalanine with a higher enantioselectivity of about 7. A colloid templated with L-lactate was applied to surface chirality analysis through enantioselective adsorption on cysteine-modified gold surfaces. Quartz microbalance experiments and scanning electron microscope observation of the gold surface revealed that the colloidal particle has higher affinity to a surface modified with L-cysteine than to one modified with D-cysteine.
Subject(s)
Amino Acids/analysis , Animals , Colloids , Electrochemistry/methods , Hydrogen-Ion Concentration , Isomerism , Lactic Acid , Microscopy, Electron, Scanning , Polymers , PyrrolesABSTRACT
PURPOSE: To determine whether secondary MR changes occur in the thalamus or the substantia nigra after middle cerebral artery (MCA) occlusion in rats. METHODS: Sprague-Dawley rats were subjected to MCA occlusion. At varying intervals, proton density-, T1-, and T2-weighted images were obtained with a 4.7-T superconductive MR unit. RESULTS: T2-weighted images revealed an area of high signal intensity in the ipsilateral substantia nigra 4 days after occlusion. A lesion of low signal intensity appeared in the ipsilateral thalamus 7 days after surgery on proton density- and/or T2-weighted images. CONCLUSION: MR showed secondary changes in the thalamus and the substantia nigra after MCA occlusion in rats. MR imaging should provide more information on the neuropathology of the delayed neuronal degeneration after cerebral ischemia.
Subject(s)
Arterial Occlusive Diseases/diagnosis , Cerebral Angiography , Cerebral Arteries/diagnostic imaging , Cerebral Infarction/diagnosis , Magnetic Resonance Imaging , Animals , Arterial Occlusive Diseases/etiology , Cerebral Arteries/pathology , Cerebral Infarction/complications , Diagnosis, Differential , Disease Models, Animal , Male , Rats , Rats, Sprague-Dawley , Sensitivity and Specificity , Substantia Nigra/diagnostic imaging , Substantia Nigra/pathology , Thalamus/diagnostic imaging , Thalamus/pathology , Tomography, X-Ray Computed , Wallerian DegenerationABSTRACT
Many hemodialysis patients are still suffering from secondary hyperparathyroidism although 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) has been used to treat renal osteodystrophy for the last two decades. The main reason for its failure to correct the secondary hyperparathyroidism is that in patients, hypercalcemia occurs before adequate parathyroid hormone (PTH) suppression is obtained when a large daily dose of 1,25(OH)2D3 is started. In this study, the oral dose of 1,25(OH)2D3 (4.0 micrograms) was administered only twice a week at the end of hemodialysis ('oral 1,25(OH)2D3 pulse therapy'), in 19 patients with severe secondary hyperparathyroidism. Serum immunoreactive PTH started to decrease after 6 weeks of therapy, and the original level of 41.2 +/- 7.24 was reduced to 24.4 +/- 6.12 ng/ml by the end of the 6-month therapy (p less than 0.001). Serum alkaline phosphatase also was reduced by 64.4%. Three out of 19 patients suffered from hypercalcemia during the 4th month of therapy. Calcium supplement given to 6 other patients with severe secondary hyperparathyroidism did not lower serum PTH levels significantly after 6 weeks of therapy, although serum calcium levels increased and were sustained above 10 mg/dl for the last 5 weeks. These findings strongly suggest that the suppressive effect of the oral 1,25(OH)2D3 pulse therapy was attained by a direct action of 1,25(OH)2D3 on the parathyroid gland rather than by its ability to elevate serum calcium levels. In conclusion, the oral 1,25(OH)2D3 pulse therapy effectively lowered PTH levels in hemodialysis patients who cannot tolerate large daily doses of 1,25(OH)2D3.
Subject(s)
Calcitriol/therapeutic use , Hyperparathyroidism, Secondary/drug therapy , Administration, Oral , Alkaline Phosphatase/blood , Calcitriol/administration & dosage , Calcitriol/blood , Calcium/administration & dosage , Calcium/blood , Dose-Response Relationship, Drug , Humans , Kidney Diseases/therapy , Parathyroid Hormone/blood , Renal DialysisABSTRACT
The toxicity/carcinogenicity of monosodium succinate, a food additive, was examined in F344 rats. The oral LD50 was greater than 8 g/kg body weight. In a 13-wk subchronic oral toxicity study, the only toxicological finding was suppression of body-weight gain in groups given greater than or equal to 2.5% monosodium succinate in the drinking-water. Histological examination revealed no toxic lesions specifically caused by the compound in any organs of any of the treated rats. The maximum tolerated dose was determined to be 2-2.5% on the basis of body-weight depression. In a long-term (2-yr) toxicity/carcinogenicity study, monosodium succinate was given ad lib. in drinking-water (distilled water) at levels of 0, 1 or 2% to groups of 50 male and 50 female rats. No toxic lesion specifically caused by long-term administration of monosodium succinate was detected. No dose-related increase was found in the incidences of tumours in any organ or tissue except for C-cell tumours of the thyroid gland of females. The incidence of these tumours in females given the 2% dose was higher than that in controls but not significantly so, and a positive trend for this tumour was noted in females. C-Cell tumour is one of the most commonly observed spontaneous tumours in ageing female rats of this strain and occurs at a variable incidence. There was no difference between the female control and treated groups in the incidence of preneoplastic change of the thyroid gland. Furthermore, the incidence of C-cell tumours in the female control group was lower than that in our historical controls. It is concluded that the increase in C-cell tumours in the female high-dose group and the detection of a positive trend for this tumour in females were probably a function of experimental variability and were not related to treatment. The results indicate that monosodium succinate had neither toxic nor carcinogenic activity in F344 rats when it was given continuously at levels of 1 or 2% in the drinking-water for 2 yr.