Angiogenesis-Inhibitory Piperidine Alkaloids from the Leaves of Microcos paniculata.

Eight new 2,6-disubstituted piperidin-3-ol alkaloids (1-8), featuring a C10 unsaturated alkyl side chain, together with three previously reported analogues (9-11) were isolated from the leaves of medicinal plant Microcos paniculata. Their structures and absolute configurations were elucidated unambiguously by means of 1D and 2D NMR spectroscopic data analysis, modified Mosher's method, Snatzke's method, and quantum chemical electronic circular dichroism (ECD) calculations, as well as single-crystal X-ray crystallography. The isolates were evaluated for their antiangiogenic effects on human umbilical vein endothelial cells (HUVECs). Compound 2 displayed an inhibitory effect on tube formation of HUVECs in a concentration-dependent manner.

Correction to: Description of Wenzhouxiangella salilacus sp. nov., a moderate halophilic bacterium isolated from a salt lake in Xinjiang Province, China.

In the original publication of the article, the deposit accession numbers of strain 15181T in the acknowledgment section were incorrectly provided as "KCTC 62172T and MCCC 1K03442T".

Anaerophilus nitritogenes gen. nov., sp. nov., isolated from salt lake sediment in Xinjiang Province, China.

An obligately anaerobic, nitrate-reducing bacterial strain (MJB2T) was isolated from sediments of saline in Xinjiang province of China. Cells were Gram-stain-positive rods and motile by means of flagella and formed endospores. The novel strain MJB2T was able to grow at 15-37 °C (optimum 28-30 °C), pH 5.8-9.4 (optimum 7.8) and with 1.0-7.0% NaCl (optimum 5.0-6.0%, w/v). Sulfate, sulfite, thiosulfate, elemental sulfur, nitrite and Fe(III) were not used as terminal electron acceptors. Oxidase and catalase reactions were positive. H2S was producted from L-cystine. Complex substrates such as beef extract, peptone and yeast extract can be used as sole energy sources. The DNA G+C content was 29.4 mol%. The major cellular fatty acids (> 10%) were C14:0, C16:1 cis 7 and C16:1 cis 9. The main polar lipids consisted of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, three unidentified amino lipids, one unidentified amino glycolipid, two unidentified glycolipid, one unidentified aminophospholipid and two unidentified lipids. No respiratory quinones were detected. According to phylogenetic analysis based on 16S rRNA gene sequences, strain MJB2T was affiliated to the family Clostridiaceae (order Clostridiales) with highest 16S rRNA gene sequence similarity of 95.3% to Crassaminicella profunda Ra1766HT. Strain MJB2T exhibited 74.9% ANI values to C. profunda Ra1766HT. In silico DNA-DNA relatedness value between strain MJB2T and C. profunda Ra1766HT was 19.5%. The distinct biochemical, chemotaxonomic and phylogenetic differences from the previously described taxa supported that strain MJB2T represents a novel species of a new genus, for which the name Anaerophilus nitritogenes gen. nov., sp. nov. is proposed. The type strain is MJB2T (=KCTC 15800T=MCCC 1K03631T).

Description of Wenzhouxiangella salilacus sp. nov., a moderate halophilic bacterium isolated from a salt lake in Xinjiang Province, China.

A Gram-stain negative, non-motile, strictly aerobic and rod-shaped bacterium, designated as 15181T, was isolated from a salt lake in Xinjiang Province, China. Strain 15181T was able to grow at 10-40 °C (optimum 37 °C), pH 6.0-8.5 (optimum 7.0) and with 1-14% NaCl (optimum 4%, w/v). According to phylogenetic analysis based on 16S rRNA gene sequences, strain 15181T was assigned to the genus Wenzhouxiangella with high 16S rRNA gene sequence similarity of 97.4% to Wenzhouxiangella sediminis XDB06T, followed by Wenzhouxiangella marina KCTC 42284T (95.9%). Strain 15181T exhibited ANI values of 80.0% and 72.0% to W. sediminis XDB06T and W. marina KCTC 42284T, respectively. The in silico DDH analysis revealed that strain 15181T shared 19.1% and 18.7% DNA relatedness with W. sediminis XDB06T and W. marina KCTC 42284T, respectively. Chemotaxonomic analysis showed that the sole respiratory quinone was ubiquinone-8, the major fatty acids included iso-C15:0, iso-C16:0 and summed feature 9 (C16:0 10-methyl and/or iso-C17:1ω9c). The major polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified glycolipids, two unidentified phospholipids, two unidentified aminophospholipids and an unidentified lipid. On the basis of phenotypic, genotypic and chemotaxonomic characteristics presented in this study, strain 15181T is concluded to represent a novel species in the genus Wenzhouxiangella, for which the name Wenzhouxiangella salilacus sp. nov. is proposed. The type strain is 15181T (=KCTC 62172T=MCCC 1K03442T).

Gallaecimonas mangrovi sp. nov., a novel bacterium isolated from mangrove sediment.

A Gram-stain negative, rod-shaped, non-motile, strictly aerobic bacterium HK-28T was isolated from a mangrove sediment sample in Haikou city, Hainan Province, China. Strain HK-28T was able to grow at 10-45 °C (optimum 25-30 °C), pH 5.0-8.5 (optimum 6.0-7.0) and 0.5-12.0% (w/v) NaCl (optimum 1.0-3.0%, w/v). The major cellular fatty acids were C16:0, Summed Feature 8 (C18:1 ω7c and/or C18:1 ω6c), Summed Feature 3 (C16:1 ω7c and/or C16:1 ω6c), C17:0, C12:0 3-OH and C17:1ω8c. Ubiquinone-8 (Q-8) was the predominant respiratory quinone. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified aminophospholipids, four unidentified phospholipids, two unidentified glycolipid, an unidentified glycophospholipid, an unidentified aminolipid and an unidentified lipid. The DNA G+C content was 50.2 mol%. Accoroding to 16S rRNA gene sequence similarities, strain HK-28T shared 97.1 and 96.7% sequence similarities to the validly named species Gallaecimonas xiamenensis MCCC 1A01354T and Gallaecimonas pentaromativorans MCCC 1A06435T, respectively, and shared lower sequence similarities (< 92.0%) to all other genera. Phylogenetic analysis showed strain HK-28T was clustered with G. pentaromativorans MCCC 1A06435T and G. xiamenensis MCCC 1A01354T. Strain HK-28T showed low DNA-DNA relatedness with G. xiamenensis MCCC 1A01354T (28.3 ± 1.5%) and G. pentaromativorans MCCC 1A06435T (25.2 ± 2.4%). On the basis of phenotypic, chemotaxonomic and genotypic characteristics, strain HK-28T is considered to represent a novel species in the genus Gallaecimonas, for which the name Gallaecimonas mangrovi sp. nov. is proposed. The type strain is HK-28T (= KCTC 62177T = MCCC 1K03441).

Muriicola marianensis sp. nov., isolated from seawater.

A Gram-stain-negative, aerobic, orange-pigmented, rod-shaped and non-motile bacterium, designated strain A6B8(T), was isolated from seawater of the Mariana Trench. The isolate grew at 4-50 °C (optimum 30-35 °C), at pH 6.5-8.0 (optimum pH 7.5) and with 0.5-4.0 % (w/v) NaCl (optimum 1.0-2.0 %). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain A6B8(T) was related most closely to the genus Muriicola and shared highest sequence similarity of 97.7 % with Muriicola jejuensis EM44(T). Chemotaxonomic analysis showed menaquinone 6 (MK-6) was the predominant isoprenoid and iso-C15 : 0, iso-C15 : 1 G and iso-C17 : 0 3-OH were the major cellular fatty acids. The polar lipid profile of strain A6B8(T) included phosphatidylethanolamine, three unidentified aminolipids and four unidentified lipids. The genomic DNA G+C content was 47.1 mol%. The DNA-DNA relatedness value (23.3 %) clearly demonstrated that strains A6B8(T) and M. jejuensis EM44(T) were representatives of two different species. Based on the phenotypic, phylogenetic and chemotaxonomic characterizations, A6B8(T) ( = CGMCC 1.12606(T) = KCTC 32436(T)) is considered to be the type strain of a novel species of the genus Muriicola, for which the name Muriicola marianensis sp. nov. is proposed.

Aquibacillus salifodinae sp. nov., a novel bacterium isolated from a salt mine in Xinjiang province, China.

A Gram-positive, rod-shaped, strictly aerobic bacterium, strain WSY08-1(T), was isolated from a salt mine in Wensu county, Xinjiang province, China. Spherical to ellipsoidal endospores were observed to be formed in terminal swollen sporangia. Strain WSY08-1(T) was found to be able to grow at 20-45 °C (optimum 37 °C), 0-10 % (w/v) NaCl (optimum 4 %, w/v) and pH 6.0-9.0 (optimum 7.0). Catalase and oxidase activities were observed to be positive. The cell-wall peptidoglycan of strain WSY08-1(T) was found to contain meso-diaminopimelic acid. Menaquinone-7 (MK-7) was identified as the predominant isoprenoid quinone. The polar lipids were found to consist of phosphatidylglycerol, diphosphatidylglycerol, an unknown glycolipid, two unknown phospholipids and an unknown lipid. The major cellular fatty acids were identified as anteiso-C15:0 and anteiso-C17:0. The DNA G+C content was determined to be 36.9 mol%. Analysis of the 16S rRNA gene sequence showed that strain WSY08-1(T) is closely related to Aquibacillus halophilus B6B(T), Aquibacillus koreensis BH30097(T) and Aquibacillus albus YIM 93642(T) (97.6, 96.9 and 96.5 % similarity, respectively). The level of DNA-DNA relatedness between strains WSY08-1(T) and A. halophilus B6B(T) was 31.4 %. On the basis of its phenotypic, chemotaxonomic and genotypic characteristics, strain WSY08-1(T) is considered to represent a novel species in the genus Aquibacillus, for which the name Aquibacillus salifodinae sp. nov. is proposed. The type strain is WSY08-1(T) (=JCM 19761(T) = CGMCC 1.12849(T)).

Bacillus salitolerans sp. nov., a novel bacterium isolated from a salt mine in Xinjiang province, China.

A novel aerobic bacterium, KC1(T), was isolated from a salt mine in Kuche county, Xinjiang province, China. Cells were observed to be Gram-positive, rod-shaped, endospore-forming and motile with flagella. Strain KC1(T) was found to grow at 25-45 °C (optimum 37 °C), pH 6.5-9.0 (optimum 8.0) and NaCl 0-10 % (v/v) (optimum 4 %). The major fatty acids were identified as anteiso-C15:0 and anteiso-C17:0. Menaquinone-7 (MK-7) was found to be the predominant isoprenoid quinone. The cell-wall diamino acid was found to be meso-diaminopimelic acid. Polar lipid analysis revealed the presence of phosphatidylglycerol and a glycolipid. The 16S rRNA gene sequence of strain KC1(T) showed low similarity (<96 %) to other validly named species. The phylogenetic trees showed that strain KC1(T) is closely related to Bacillus azotoformans DSM 1046(T) and Bacillus methanolicus DSM 16454(T). Both these type strains showed 95.4 % 16S rRNA gene sequence similarity to strain KC1(T). The DNA G+C content of strain KC1(T) was determined to be 39.0 mol%. On the basis of its phenotypic, chemotaxonomic and genotypic characteristics, strain KC1(T) is considered to represent a novel species of the genus Bacillus, for which the name Bacillus salitolerans sp. nov. is proposed. The type strain is KC1(T) (=JCM 19760(T) = CGMCC 1.12810(T)).

Devosia pacifica sp. nov., isolated from deep-sea sediment.

A novel bacterial strain, NH131(T), was isolated from deep-sea sediment of South China Sea. Cells were strictly aerobic, Gram-stain negative, short rod-shaped and motile with a single lateral flagellum. Strain NH131(T) grew optimally at pH 6.5-7.0 and 25-30 °C. 16S rRNA gene sequence analysis revealed that strain NH131(T) belonged to the genus Devosia, sharing the highest sequence similarity with the type strain, Devosia geojensis BD-c194(T) (96.2%). The predominant fatty acids were C(18 : 1)ω7c, 11-methyl C(18 : 1)ω7c, C(18 : 0) and C(16 : 0). Ubiquinone 10 was the predominant ubiquinone. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phospholipid, three glycolipids and two unknown lipids. The DNA G+C content of strain NH131(T) was 63.0 mol%. On the basis of the results of polyphasic identification, it is suggested that strain NH131(T) represents a novel species of the genus Devosia for which the name Devosia pacifica sp. nov. is proposed. The type strain is NH131(T) ( = JCM 19305(T) = KCTC 32437(T)).

Halopiger salifodinae sp. nov., an extremely halophilic archaeon isolated from a salt mine.

A novel extremely halophilic archaeon KCY07-B2(T) was isolated from a salt mine in Kuche county, Xinjiang province, China. Colonies were cream-pigmented and cells were pleomorphic rod-shaped. Strain KCY07-B2(T) was able to grow at 25-50 °C (optimum 37-45 °C) and pH 6.0-8.0 (optimum 7.0). The strain required at least 1.9 M NaCl for growth. MgCl2 was not required. Cells lysed in distilled water. Polar lipid analysis revealed the presence of phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester, derived from both C20C20 and C20C25 glycerol diethers, together with five glyolipids. The bis-sulfated glycolipid S2-DGD-1 was present. The DNA G+C content was 62.5 mol%. Analysis of the 16S rRNA gene sequence revealed that strain KCY07-B2(T) was closely related to Halopiger xanaduensis SH-6(T) and Halopiger aswanensis 56(T) (95.8 % and 95.5 % similarity, respectively). On the basis of its phenotypic, chemotaxonomic and genotypic characteristics, strain KCY07-B2(T) is considered to represent a novel species of the genus Halopiger, for which the name Halopiger salifodinae sp. nov. is proposed. The type strain is KCY07-B2(T) ( = JCM 18547(T) = CGMCC 1.12284(T)).

Halopelagius longus sp. nov., a member of the family Halobacteriaceae isolated from a salt mine, and emended description of the genus Halopelagius.

A thermotolerant, extremely halophilic archaeon, BC12-B1(T), was isolated from a salt mine in Baicheng county, Xinjiang province, China. Colonies were off-white-grey. The cells stained Gram-negative, were motile and irregularly long-rod-shaped (variation in both width and length) with abundant gas vesicles. The strain was able to grow at 20-55 °C (optimum, 48 °C), at pH 6.0-8.0 (optimum, 7.0-7.3), with 1.8-6.0 M NaCl (optimum, 3.0-3.5 M) and with 0.02-2.2 M Mg(2+) (optimum, 0.1-0.2 M). Cells lysed in distilled water and the minimal NaCl concentration to prevent cell lysis was 8 % (w/v). Phylogenetic analysis based on the 16S rRNA gene sequences showed that strain BC12-B1(T) was most closely related to Halopelagius inordinatus RO5-2(T) (98.5 %) with less than 95 % sequence similarity to other described species. The genomic DNA G+C content of strain BC12-B1(T) was 64.0 mol%. The DNA-DNA hybridization value between strain BC12-B1(T) and Hpl. inordinatus RO5-2(T) was 43.6 %. The major polar lipids of strain BC12-B1(T) were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, four glycolipids and an unknown lipid. Based on phenotypic, chemotaxonomic and genotypic characteristics, strain BC12-B1(T) represents a novel species of the genus Halopelagius, for which the name Halopelagius longus sp. nov. is proposed. The type strain is BC12-B1(T) ( = CGMCC 1.12397(T) = JCM 18758(T)). An emended description of the genus Halopelagius is also provided.

Halolamina salifodinae sp. nov. and Halolamina salina sp. nov., two extremely halophilic archaea isolated from a salt mine.

Two strictly aerobic, extremely halophilic archaea, strains WSY15-H1(T) and WSY15-H3(T), were isolated from a salt mine in Wensu county, Xinjiang province, China. Cells of the two strains were Gram-stain-negative, non-motile and pleomorphic. Colonies were pink- and red-pigmented, respectively. Strain WSY15-H1(T) grew at 20-45 °C (optimum 37-42 °C), 1.6-5.4 M NaCl (optimum 3.4-3.9 M), 0-2.0 M MgCl2 (optimum 0.1-0.5 M) and pH 6.0-9.0 (optimum 7.0), whereas strain WSY15-H3(T) grew at 20-50 °C (optimum 37 °C), 1.9-5.4 M NaCl (optimum 3.4 M), 0.02-2.5 M MgCl2 (optimum 0.5-1.0 M) and pH 6.0-7.5 (optimum 6.5). The minimal NaCl concentrations to prevent cell lysis were 9 % (w/v) for strain WSY15-H1(T) and 8 % (w/v) for strain WSY15-H3(T). The major polar lipids of the two isolates were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and phosphatidylglycerol sulfate, as well as nine glycolipids for strain WSY15-H1(T) and seven glycolipids for strain WSY15-H3(T); two of these glycolipids (GL1 and GL3) were chromatographically identical to bis-sulfated diglycosyl diether (S2-DGD-1) and sulfated diglycosyl diether (S-DGD-1), respectively. The genomic DNA G+C contents of strains WSY15-H1(T) and WSY15-H3(T) were 65.4 and 66.2 mol%. On the basis of 16S rRNA gene sequence analysis, strains WSY15-H1(T) and WSY15-H3(T) shared 97.0% similarity with each other and showed respectively 98.4 and 97.6% sequence similarity to Halolamina pelagica TBN21(T), which was the only type strain that had higher than 91% sequence similarity with the two isolates. Analysis of phylogenetic relationships and DNA-DNA relatedness indicated that strains WSY15-H1(T) and WSY15-H3(T) represent two novel lineages with closest affinity to H. pelagica TBN21(T). Based on phenotypic, chemotaxonomic and genotypic characteristics, two novel species of the genus Halolamina are proposed, Halolamina salifodinae sp. nov. (type strain WSY15-H1(T) = JCM 18548(T) = GCMCC 1.12371(T)) and Halolamina salina sp. nov. (type strain WSY15-H3(T) = JCM 18549(T) = GCMCC 1.12285(T)).

Extensimonas vulgaris gen. nov., sp. nov., a member of the family Comamonadaceae.

A novel strain, named S4(T), was obtained from industrial wastewater in Xiaoshan, Zhejiang Province, China. Cells were Gram-negative, neutrophilic and non-spore-forming and moved by means of a polar flagellum. Normal cells were 0.8-0.9 × 1.3-1.9 µm and the cells elongated to 10-25 µm when cultivated at high temperatures. Strain S4(T) grew at 15-50 °C (optimum at 48 °C), pH 5.5-8.5 (optimum 7.0-7.5) and 0-2% (optimum 0.5%) (w/v) NaCl. Ubiquinone-8 was the predominant respiratory quinone. C16:0, summed feature 3 (C16:1ω7c and/or iso-C15:0 2-OH) and C17:0 cyclo were the major cellular fatty acids. The major 3-OH fatty acid was C10:0 3-OH. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unknown aminoglycolipid. The genomic DNA G+C content was 68.8 mol%. Based on 16S rRNA gene sequences alignment, the most closely related strains were members of the genera Comamonas (94.6-95.6% similarities), Giesbergeria (94.9-95.6%), Acidovorax (94.8-95.4%), Brachymonas (94.1-95.2%) and Macromonas (95.1%). Phylogenetic analysis showed the closest relatives of strain S4(T) were members of the genus Macromonas. Based on phenotypic and phylogenetic characteristics, we suggest that strain S4(T) represents a novel species of a new genus of the family Comamonadaceae, for which the name Extensimonas vulgaris gen. nov., sp. nov. is proposed. The type strain of Extensimonas vulgaris is S4(T) (=CGMCC 1.10977(T)=JCM 17803(T)).

Prognostic value of magnetic resonance imaging in patients with clinically isolated syndrome conversion to multiple sclerosis: a meta-analysis.

BACKGROUND: Numerous studies have investigated the associations of brain or spinal cord MRI with the risk of developing Multiple Sclerosis (MS) in people with Clinically Isolated Syndrome (CIS), however, the findings are uncertain. Therefore, we performed a meta-analysis based on 24 publications to comprehensively evaluate such associations. MATERIALS AND METHODS: The databases of EMBASE and MEDLINE (January 1980-August 2011) were searched electronically for all relevant studies. Data were extracted from each study independently by both reviewers using a predefined structured spreadsheet. The quality of each study was assessed independently by two reviewers according to Newcastle-Ottawa Scale for reading cohort study proposed by Deeks et al. The meta-analysis including 24 qualified studies was performed by using the Cochrane Collaborations RevMan5.0 software. RESULTS: Twenty-four identified studies met the inclusion criteria and minimum quality threshold. A meta-analysis of cohort studies indicated that the CISs having MRI lesions did have significantly increased risk for MS (risk ratio [RR] = 3.71, 95% confidence interval [CI], 3.27-4.21, P < 0.00001). In the subgroup analysis (according to the number of T2 lesions at baseline), the risk of developing MS in CIS patients with the medium MRI burden (4-9 lesions) was higher than with the low MRI burden (1-3 lesions) (RR = 0.66,95% CI, 0.45-0.95, P < 0.00001). While, no correlation was found in group between the medium MRI burden and the high MRI burden(>9 lesions) (RR = 0.97, 95% CI, 0.82-1.15, P = 0.72). Meanwhile, the CIS patients with abnormal baseline MRI, especially with infratentorial lesions, had a high risk of conversion to MS compared to patients without the such infratentorial lesions (RR = 1.37, % CI, 1.09-1.73, P = 0.0008). CONCLUSIONS: Despite some limitations, this meta-analysis established solid statistical evidence for an association between the presence or absence of MRI lesions within the brain or spinal cord MRI and the risk of developing MS, particularly for studies with large sample size. The CIS patients with abnormal baseline MRI, especially with infratentorial lesions, had a high risk of conversion to MS. However, this association warrants additional validation in larger and well designed studies.

[Investigation of amylin level changes among obese patients at different glucose metabolic states and effects of rosiglitazone intervention].

OBJECTIVE: To measure the changes in plasma amylin level among obese patients at different glucose metabolic states, and to evaluate effects of rosiglitazone intervention on obese type 2 diabetes patients. METHODS: A total of 92 obese patients were categorized into normal glucose tolerance group (Group A, n=31), impaired glucose tolerance group (Group B, n=30), and type 2 diabetes group (Group C, n=31) according to oral glucose tolerance test (OGTT) results. Within the new type 2 diabetes group, patients were further randomized into 4 mg rosiglitazone treatment group and life style adjustment group. Body mass index (BMI) and waist circumference of all the patients were measured, and their plasma amylin and true insulin levels measured by radioimmunoassay and EIA. RESULTS: Compared with Group A, both fasting and 30 minute glucose load plasma amylin levels, and ΔAmylin30/ΔGlucose30 in Group B and C were lower. Compared with the life style adjustment group, both fasting and 30 minute plasma amylin levels, and homeostasis model assessment for B cell function (HOMA-B) were higher in the group that received rosiglitazone treatment, but still lower than those in the Group A. CONCLUSION: Pancreatic B cell function and amylin secretion were impaired in the abnormal metabolic states of impaired glucose tolerance and type 2 diabetes patients. Rosiglitazone intervention helped to improve B cell function and increase amylin level.

Absolute Configurations and Stereochemical Inversion Mechanism of Epimeric Securinega Alkaloids from Flueggea suffruticosa.

Three pairs of Securinega alkaloid epimers with a piperidin-2-yl moiety (1-6) were isolated from Flueggea suffruticosa, and their structures including absolute configurations were definitely characterized. An interconvertible C-2' epimerization process within each pair of epimers was observed. The following comprehensive experimental and theoretical investigations demonstrated an unusual stereochemical inversion mechanism of an N-substituted carbon stereogenic center, which was evidenced to be a protic solvent mediated process involving a tandem 1,4-elimination/1,4-addition as the key step.

Alkaloid constituents from the fruits of Flueggea virosa.

Three new indole alkaloids, flueindolines A-C (1-3), along with nine known alkaloids (4-12), were isolated from the fruits of Flueggea virosa (Roxb. ex Willd.) Voigt. Compounds 1 and 2 are two new fused tricyclic indole alkaloids possessing an unusual pyrido[1, 2-a]indole framework, and 3 presents a rare spiro (pyrrolizidinyl-oxindole) backbone. Their structures with absolute configurations were elucidated by means of comprehensive spectroscopic analysis, chemical calculation, as well as X-ray crystallography. Chiral resolution and absolute configuration determination of the known compounds 4, 10, and 11 were reported for the first time. The hypothetical biogenetical pathways of 1-3 were herein also proposed.

Expression and functional analysis of CYP2D6.24, CYP2D6.26, CYP2D6.27, and CYP2D7 isozymes.

The objectives of this study were to compare the drug-metabolizing activity of human CYP2D6.24 (I297L), CYP2D6.26 (I369T), and CYP2D6.27 (E410K) allelic isoforms with wild-type CYP2D6.1 and to express the CYP2D7 protein derived from an indel polymorphism (CYP2D7 138delT) and investigate its possible codeine O-demethylase activity. Successful creation of individual cDNAs corresponding to CYP2D6*24 (2853 A>C), CYP2D6*26 (3277 T>C), and CYP2D6*27 (3853 G>A) allelic variants and CYP2D7 was achieved via molecular cloning. The corresponding proteins, CYP2D6.24, CYP2D6.26, CYP2D6.27, and CYP2D7, were expressed in insect cells by using a baculovirus-mediated expression system. All CYP2D proteins showed the empirical carbon monoxide difference spectra. We were surprised to find that the CYP2D7 protein was detected mainly in mitochondrial fractions, whereas all CYP2D6 allelic isoforms were present in the microsomal fraction. Furthermore, CYP2D7 did not produce any morphine from codeine. In contrast, CYP2D6.24, CYP2D6.26, and CYP2D6.27 allelic isoforms all showed active drug-metabolizing activities toward both codeine and dextromethorphan O-demethylation. Whereas CYP2D6.24 exhibited the highest intrinsic clearance in dextromethorphan O-demethylation (approximately 6-fold higher than that by CYP2D6.1), it had the lowest enzyme efficiency in codeine O-demethylation (approximately 50% lower than that by CYP2D6.1). Overall, the enzymatic consequences of CYP2D6 allelic isozymes are substrate dependent. These data would help preclinical and clinical assessments of the metabolic elimination of drugs that are mediated by human CYP2D enzyme.

The Succinate Receptor GPR91 Is Involved in Pressure Overload-Induced Ventricular Hypertrophy.

BACKGROUND: Pulmonary arterial hypertension is characterized by increased pressure overload that leads to right ventricular hypertrophy (RVH). GPR91 is a formerly orphan G-protein-coupled receptor (GPCR) that has been characterized as a receptor for succinate; however, its role in RVH remains unknown. METHODS AND RESULTS: We investigated the role of succinate-GPR91 signaling in a pulmonary arterial banding (PAB) model of RVH induced by pressure overload in SD rats. GPR91 was shown to be located in cardiomyocytes. In the sham and PAB rats, succinate treatment further aggravated RVH, up-regulated RVH-associated genes and increased p-Akt/t-Akt levels in vivo. In vitro, succinate treatment up-regulated the levels of the hypertrophic gene marker anp and p-Akt/t-Akt in cardiomyocytes. All these effects were inhibited by the PI3K antagonist wortmannin both in vivo and in vitro. Finally, we noted that the GPR91-PI3K/Akt axis was also up-regulated compared to that in human RVH. CONCLUSIONS: Our findings indicate that succinate-GPR91 signaling may be involved in RVH via PI3K/Akt signaling in vivo and in vitro. Therefore, GPR91 may be a novel therapeutic target for treating pressure overload-induced RVH.