Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 20
Filter
1.
Ecol Evol ; 13(5): e10032, 2023 May.
Article in English | MEDLINE | ID: mdl-37153019

ABSTRACT

The original description of Natrix leonardi (currently Rhabdophis leonardi) by Frank Wall in 1923, based on a specimen from the "Upper Burma Hills," lacked important morphological details that have complicated the assignment of recently collected material. Furthermore, although the holotype was never lost, its location has been misreported in one important taxonomic reference, leading to further confusion. We report the correct repository of the holotype (Natural History Museum, London), together with its current catalog number. We also describe key features of that specimen that were omitted from the original description, and provide new details on the morphology of the species, including sexual dichromatism unusual for the genus, based upon specimens from southern Sichuan, China. Rhabdophis leonardi is distinguished from its congeners by the following characters: 15 or 17 DSR at midbody and 6 supralabials; distinct annulus around the neck, broad and red in males, and narrow and orange with a black border in females; dorsal ground color light green or olive; some lateral and dorsal scales possessing black edges, the frequency of black edges gradually increasing from anterior to posterior, forming irregular and ill-defined transverse black bands; eye with prominent green iris; black ventral spots with a red edge, most numerous at midbody but extending halfway down the length of the tail. In southwestern China, this species is frequently found at 1730-2230 m elevation. It has been documented to prey upon anuran amphibians, including toads. A recently published phylogenetic analysis showed this species to be deeply nested with the genus Rhabdophis, as a member of the R. nuchalis Group. That analysis also revealed the existence of two closely related but geographically distinct subclades in the molecular analysis, one of which may represent an unnamed taxon.

2.
BMC Nephrol ; 23(1): 49, 2022 01 29.
Article in English | MEDLINE | ID: mdl-35093023

ABSTRACT

BACKGROUND: The urine protein-creatinine ratio (UPCR) in a spot first-morning urine sample is used to estimate 24-h urine proteinuria (24hUP) in patients who underwent urine protein testing. UPCR cannot be directly compared with 24-h proteinuria. Thus, an equation to estimate 24-h total protein excretion rate, using age, gender, and the UPCR may improve its bias and accuracy in patients who underwent urine protein testing. METHODS: We simultaneously measured 24-h urine protein and the same day's first-morning spot urine from patients with kidney disease. Generalized linear and no-linear models, using age, gender, and UPCR, were constructed to estimate for 24-h urine protein and the best model (NJ equation) was selected to estimated 24 hUP (e24hUP). RESULTS: A total of 5435 paired samples (including a training cohort of 3803 patients and a validation cohort of 1632 patients) were simultaneously measured for UPCR and 24-h urine protein. In the training cohort, the unadjusted UPCR obviously underestimated 24-h urine protein when UPCR ≤1.2 g/g (median bias - 0.17 g/24 h) and overestimated 24-h urine protein when UPCR > 1.2 g/g (median bias 0.53 g/24 h). In the validation cohort, the NJ equation performed better than the unadjusted UPCR, with lower root mean square error (0.81 vs. 1.02, P < 0.001), less bias (median difference between measured and estimated urine protein, - 0.008 vs. 0.12), improved precision (interquartile range of the differences, 0.34 vs. 0.50), and greater accuracy (percentage of estimated urine protein within 30% of measured urine protein, 53.4% vs. 32.2%). Bland-Altman plot indicated that the agreement of spot and daily estimates was less pronounced with 24 hUP > 2 g than lower values. CONCLUSIONS: The NJ e24hUP equation is more accurate than unadjusted UPCR to estimate 24 hUP in patients with kidney disease and could be used for laboratory application.


Subject(s)
Creatinine/urine , Proteinuria/urine , Adult , Female , Humans , Male , Mathematical Concepts , Middle Aged , Monitoring, Physiologic , Urinalysis/methods
3.
Zookeys ; 1114: 59-76, 2022.
Article in English | MEDLINE | ID: mdl-36761701

ABSTRACT

This study presents a comprehensive morphological comparison along with molecular phylogeny of the genus Gloydius based on five mitochondrial genes (12S, 16S, COI, cytb, and ND4). The specimens collected from Jiuzhaigou National Nature Reserve are shown to be a new species, Gloydiuslateralis sp. nov. Zhang, Shi, Jiang & Shi based on a combination of morphological and molecular accounts. G.lateralis sp. nov. differs from other congeneric species by a series of diagnostic morphological characteristics and forms a strongly supported monophyletic group. The new species is phylogenetically closely related to G.swild, another recently described species from Heishui, Aba, Sichuan.

4.
Zookeys ; 1061: 87-108, 2021.
Article in English | MEDLINE | ID: mdl-34707454

ABSTRACT

We provide a molecular phylogeny of Asian pit vipers (the genus Gloydius) based on four mitochondrial genes (12S, 16S, ND4, and cytb). Sequences of Gloydiushimalayanus, the only member of the genus that occurs south of the Himalayan range, are included for the first time. In addition, two new species of the genus Gloydius are described based on specimens collected from Zayu, Tibet, west of the Nujiang River and Heishui, Sichuan, east of the Qinghai-Tibet Plateau. The new species, Gloydiuslipipengi sp. nov., can be differentiated from its congeners by the combination of the following characters: the third supralabial not reaching the orbit (separated from it by a suborbital scale); wide, black-bordered greyish postorbital stripe extending from the posterior margin of the orbit (not separated by the postoculars, covering most of the anterior temporal scale) to the ventral surface of the neck; irregular black annular crossbands on the mid-body; 23-21-15 dorsal scales; 165 ventral scales, and 46 subcaudal scales. Gloydiusswild sp. nov. can be differentiated from its congeners by the narrower postorbital stripe (only half the width of the anterior temporal scale, the lower edge is approximately straight and bordered with white); a pair of arched stripes on the occiput; lateral body lakes black spots; a pair of round spots on the parietal scales; 21 rows of mid-body dorsal scales; zigzag dark brown stripes on the dorsum; 168-170 ventral scales, and 43-46 subcaudal scales. The molecular phylogeny in this study supports the sister relationship between G.lipipengi sp. nov. and G.rubromaculatus, another recently described species from the Qinghai-Tibet Plateau, more than 500 km away, and indicate the basal position of G.himalayanus within the genus and relatively distant relationship to its congeners.

5.
Zookeys ; 1048: 23-47, 2021.
Article in English | MEDLINE | ID: mdl-34295215

ABSTRACT

Based on combined morphological and osteological characters and molecular phylogenetics, we describe a new species of the genus Elaphe that was discovered from the south slope of the Qinling Mountains, Shaanxi, China, namely Elaphe xiphodonta sp. nov. It is distinguished from the other congeners by a combination of the following characters: dorsal scales in 21-21-17 rows, the medial 11 rows keeled; 202-204 ventral scales, 67-68 subcaudals; two preoculars (including one subpreocular); two postoculars; two anterior temporals, three posterior temporals; reduced numbers of maxillary teeth (9+2) and dentary teeth (12); sharp cutting edges on the posterior or posterolateral surface of the rear maxillary teeth and dentary teeth; dorsal head yellow, three distinct markings on the head and neck; a distinct black labial spot present in supralabials; dorsum yellow, 46-49 complete (or incomplete) large black-edged reddish brown blotches on the body and 12-19 on the tail, two rows of smaller blotches on each ventrolateral side; ventral scales yellow with mottled irregular black blotches, a few irregular small red spots dispersed on the middle of the ventral. Based on molecular phylogenetic analyses, the new species forms the sister taxon to E. zoigeensis. The discovery of this new species increases the number of the recognized species in the genus Elaphe to 17.

6.
Clin J Am Soc Nephrol ; 16(8): 1221-1227, 2021 08.
Article in English | MEDLINE | ID: mdl-34083219

ABSTRACT

BACKGROUND AND OBJECTIVES: Associations between HLA alleles and susceptibility to M-type phospholipase A2 receptor (PLA2R)-related membranous nephropathy have been well defined previously in Chinese patients. However, the relationships between HLA alleles and kidney outcome remain unclear. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Five HLA genes (DRB1, DQA1, DQB1, DRB3, and DRB5) were genotyped in a prospective cohort of 392 patients with PLA2R-related membranous nephropathy. The associations between HLA alleles and kidney outcomes were studied. RESULTS: A total of 79 HLA alleles were identified in this study. Four HLA alleles, DRB1*13:01 (n=12; hazard ratio, 3.7; 95% confidence interval, 1.8 to 7.8; P<0.001), DQB1*06:03 (n=12; hazard ratio, 3.7; 95% confidence interval, 1.8 to 7.8; P<0.001), DRB1*04:05 (n=12; hazard ratio, 3.8; 95% confidence interval, 1.5 to 9.5; P=0.004), and DQB1*03:02 (n=21; hazard ratio, 3.1; 95% confidence interval, 1.4 to 6.7; P=0.005), were associated with a ≥40% eGFR decline during follow-up. DRB1*13:01 and DQB1*06:03 were tightly linked with each other. Forty-four of the 392 patients (11%) carried at least one of the four identified risk HLA alleles in this study. Compared with patients who were negative for all risk HLA alleles, those carrying at least one risk HLA allele had a significant risk of a ≥40% eGFR decline during follow-up (hazard ratio, 3.9; 95% confidence interval, 2.3 to 6.7; P<0.001). After adjusting for age, sex, proteinuria, albumin, eGFR, and anti-PLA2R antibody levels, multivariable Cox analysis showed that patients carrying any of the four risk HLA alleles remained associated with a higher risk of a ≥40% decline in eGFR (hazard ratio, 4.1; 95% confidence interval, 2.3 to 7.1; P<0.001). CONCLUSIONS: Carrying any of the HLA alleles, DRB1*13:01/DQB1*06:03, DRB1*04:05, and DQB1*03:02, was independently associated with poor prognosis in Chinese patients with PLA2R-related membranous nephropathy.


Subject(s)
Glomerulonephritis, Membranous/genetics , HLA-DQ alpha-Chains/genetics , HLA-DQ beta-Chains/genetics , HLA-DR beta-Chains/genetics , Receptors, Phospholipase A2/genetics , Adult , Alleles , Asian People/genetics , China , Disease Progression , Female , Genotype , Glomerular Filtration Rate , Glomerulonephritis, Membranous/physiopathology , HLA-DRB1 Chains/genetics , HLA-DRB3 Chains/genetics , HLA-DRB5 Chains/genetics , Humans , Male , Middle Aged , Prognosis , Prospective Studies , Risk Factors
7.
Zookeys ; 1025: 35-71, 2021.
Article in English | MEDLINE | ID: mdl-33814945

ABSTRACT

Kraits of the genus Bungarus Daudin 1803 are widely known venomous snakes distributed from Iran to China and Indonesia. Here, we use a combination of mitochondrial DNA sequence data and morphological data to describe a new species from Yingjiang County, Yunnan Province, China: Bungarus suzhenae sp. nov. Phylogenetically, this species forms a monophyletic lineage sister to the Bungarus candidus/multicinctus/wanghaotingi complex based on cyt b and ND4 genes but forms a sister species pair with the species B. magnimaculatus Wall & Evans, 1901 based on COI gene fragments. Morphologically, B. suzhenae sp. nov. is similar to the B. candidus/multicinctus/wanghaotingi complex but differs from these taxa by a combination of dental morphology, squamation, coloration pattern, as well as hemipenial morphology. A detailed description of the cranial osteology of the new species is given based on micro-CT tomography images. We revised the morphological characters of B. candidus/multicinctus/wanghaotingi complex and verified the validity of three species in this complex. The distribution of these species was revised; the records of B. candidus in China should be attributed to B. wanghaotingi. We also provide an updated key to species of Bungarus.

9.
Chin Med J (Engl) ; 131(22): 2658-2665, 2018 Nov 20.
Article in English | MEDLINE | ID: mdl-30425192

ABSTRACT

BACKGROUND: Treatment with the dipeptidyl peptidase-4 inhibitors (DPP4i) and angiotensin receptor blockers (ARBs) in patients with type 2 diabetic nephropathy (DN) has not been well characterized. This study aimed to assess the renoprotection of this combined treatment in DN patients. METHODS: A total of 159 type 2 DN patients from 2013 to 2015 were enrolled retrospectively from a prospective DN cohort at the National Clinical Research Center of Kidney Diseases, Jinling Hospital (China). Fifty-seven patients received DPP4i and ARB treatment, and 102 patients were treated with ARBs alone. All patients were followed up for at least 12 months. Statistical analyses were performed using Stata version 12.0. RESULTS: There were no significant differences at baseline for age, sex, body mass index, duration of diabetes, fasting blood glucose (FBG), hemoglobin A1c (HbA1c), and estimated glomerular filtration rate (eGFR) between the two groups. Antihypertensive and antidiabetic medication use was similar in each group except calcium channel antagonists (P = 0.032). No significant changes in FBG and HbA1c were observed in the two groups after treatment. The eGFR decreased slower in the DPP4i + ARB group than in the ARB group at 12 months (Δ12 months: -2.48 ± 13.86 vs. -6.81 ± 12.52 ml·min-1·1.73m-2, P = 0.044). In addition, proteinuria was decreased further in the DPP4i + ARB group than in the ARB group after 24 months of treatment (Δ24 months: -0.18 [-1.00, 0.17] vs. 0.32 [-0.35, 0.88], P = 0.031). There were 36 patients with an eGFR decrease of more than 30% over 24 months. After adjusting for FBG, HbA1c, and other risk factors, DPP4i + ARB treatment was still associated with a reduced incidence of an eGFR decrease of 20% or 30%. CONCLUSIONS: The combined treatment of DPP4i and ARBs is superior to ARBs alone, as evidenced by the greater proteinuria reduction and lower eGFR decline. In addition, the renoprotection of DPP4i combined with ARBs was independent of glycemic control.


Subject(s)
Angiotensin Receptor Antagonists/therapeutic use , Diabetic Nephropathies/drug therapy , Dipeptidyl-Peptidase IV Inhibitors/therapeutic use , Aged , Female , Humans , Losartan/therapeutic use , Male , Middle Aged , Prospective Studies , Retrospective Studies
10.
BMC Nephrol ; 18(1): 2, 2017 Jan 05.
Article in English | MEDLINE | ID: mdl-28056860

ABSTRACT

BACKGROUND: The KDIGO Clinical Practice Guidelines for Glomerulonephritis recommended tacrolimus as an alternative regimen for the initial therapy for Idiopathic membranous nephropathy (IMN), however, large observational studies evaluating tacrolimus treatment in IMN remains rare. METHODS: A total of 408 consecutive IMN patients with nephrotic syndrome who were treated with tacrolimus in Jinling Hospital were included. The effectiveness and safety of tacrolimus treatment in IMN were analyzed in this study. RESULTS: The cumulative partial or complete remission after tacrolimus therapy were 50%, 63% and 67% at 6, 12 and 24 months, respectively, and the cumulative complete remission rates were 4%, 13% and 23%, respectively. Multivariate logistic analysis showed that higher tacrolimus exposure during induction treatment, female gender, higher eGFR and no history of previous immunosuppressive therapy were independently associated with higher probability of remission. A relapse occurred in 101 of the 271 (37.3%) patients with partial or complete remission, and 18 of the 95 (18.9%) patients with complete remission. Tapering duration of tacrolimus and complete remission versus partial remission status were independent factors associated with risk of relapse. A decline in eGFR was the most frequent adverse event during tacrolimus treatment. During tacrolimus treatment, a ≥40% decrease in eGFR was observed in 43 (10.5%) patients. CONCLUSIONS: Low dose tacrolimus is effective for IMN, with a total remission rate of 66% whereas with a rather high rate of relapse. However, the safety of tacrolimus treatment needs to be further validated in large randomized clinical trials.


Subject(s)
Drug-Related Side Effects and Adverse Reactions/epidemiology , Glomerulonephritis, Membranous/drug therapy , Glomerulonephritis, Membranous/epidemiology , Tacrolimus/administration & dosage , Adult , China/epidemiology , Cohort Studies , Dose-Response Relationship, Drug , Drug-Related Side Effects and Adverse Reactions/prevention & control , Female , Humans , Immunosuppressive Agents/administration & dosage , Longitudinal Studies , Male , Prevalence , Recurrence , Risk Factors , Sex Distribution , Treatment Outcome
11.
J Am Soc Nephrol ; 28(5): 1642-1650, 2017 May.
Article in English | MEDLINE | ID: mdl-28028136

ABSTRACT

Idiopathic membranous nephropathy (MN) is associated with HLA; however, the HLA allele involved remains unknown. To identify the HLA risk alleles associated with phospholipase A2 receptor (PLA2R)-related MN in the Chinese population, we sequenced the entire MHC region in DNA samples from 99 patients with PLA2R-related MN, 50 patients with PLA2R-unrelated MN, and 100 healthy subjects. Two HLA risk alleles, HLA-DRB1*15:01 and HLA-DRB3*02:02, independently and strongly associated with an increased risk of PLA2R-related MN. After adjusting for HLA-DRB1*15:01 and HLA-DRB3*02:02, no other alleles showed significant association with PLA2R-related MN. A replication study in an independent cohort of 293 participants with PLA2R-related MN and 285 healthy controls validated these findings. In a joint analysis, a multivariate logistic regression model confirmed that HLA-DRB1*15:01 (odds ratio [OR], 24.9; 95% confidence interval [95% CI], 15.3 to 42.6; P=2.3×10-35) and HLA-DRB3*02:02 (OR, 17.7; 95% CI, 11.0 to 30.3; P=8.0×10-29) independently and strongly associated with PLA2R-related MN. As many as 98.7% of patients with PLA2R-related MN, compared with 43.9% of control subjects, carried at least one HLA risk allele. Subjects with either risk allele had higher odds of developing PLA2R-related MN than those without a risk allele (OR, 98.9; 95% CI, 44.4 to 281.7; P=2.5×10-23). These HLA risk alleles also associated with the age at disease onset in patients with PLA2R-related MN. In conclusion, our findings provide clear evidence that the HLA-DRB1*15:01 and HLA-DRB3*02:02 alleles independently and strongly associate with PLA2R-related MN in the Chinese population.


Subject(s)
Glomerulonephritis, Membranous/genetics , HLA-DRB1 Chains/genetics , HLA-DRB3 Chains/genetics , Receptors, Phospholipase A2/physiology , Adult , Alleles , Asian People , Female , Glomerulonephritis, Membranous/immunology , HLA-DRB1 Chains/immunology , HLA-DRB3 Chains/immunology , Humans , Male , Middle Aged , Young Adult
12.
Exp Ther Med ; 10(1): 237-240, 2015 Jul.
Article in English | MEDLINE | ID: mdl-26170941

ABSTRACT

Patients with traumatic hemorrhagic shock are highly susceptible to the development of acute kidney injury (AKI), but little data are available regarding the changes in cystatin C (CysC) in patients with traumatic hemorrhagic shock. The aim of the present study, therefore, was to investigate whether CysC has a higher value than serum creatinine (SCr) and urea for use in monitoring glomerular function in traumatic hemorrhagic shock. Data from a cohort of patients with traumatic hemorrhagic shock, who had been admitted to a trauma center, were collected. Receiver operating characteristic (ROC) curve analysis was used to determine the diagnostic value of serum CysC, SCr and urea for the identification of renal dysfunction, and the data were expressed as the area under the curve (AUC). CysC was not significantly affected by gender, age, mechanism of injury or time between injury and arrival at the center in the patients with traumatic hemorrhagic shock. The CysC level of the patients was significantly higher than that of the normal subjects (1.10±0.36 vs. 0.91±0.34 mg/l); the SCr and urea levels of the patients were also significantly increased compared with those of the normal subjects. Nonparametric ROC plots of the sensitivity and specificity of SCr, CysC and urea for the detection of AKI revealed AUC values of 0.901 [95% confidence interval (CI), 0.791-1.000], 0.728 (95% CI, 0.570-0.886) and 0.709 (95% CI, 0.552-0.865) for SCr, CysC and urea, respectively. No significant correlation between mortality and CysC, SCr or urea was found. These data indicate that the level of CysC is significantly increased in the early stage of traumatic hemorrhagic shock and that CysC can be used as a marker to predict AKI; however, the diagnostic utility of CysC remains lower than that of SCr in the early stage of the condition.

13.
J Basic Microbiol ; 53(11): 934-41, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23457057

ABSTRACT

Nitrilase-mediated biocatalysis has attracted substantial attention for its application in carboxylic acid production in recent years. In the present study, the fungus CA3-1 was isolated and identified as Gibberella intermedia based on its morphology, its 18S ribosomal DNA (rDNA), and internal transcribed spacer (ITS) sequences. The enzymatic properties of G. intermedia resting cells were determined, and the optimum activity was achieved at 40 °C with pH 7.6. The half-lives of the nitrilase at 30, 40, and 50 °C were 231.1, 72.9, and 6.4 h, respectively. This Gibberella nitrilase showed a wide substrate spectrum with high specificity for heterocyclic and aliphatic nitriles. It remained extremely active in 5% propanol. The presence of Ag(+), Hg(2+), and excess substrate inhibited the nitrilase activity, whereas Fe(2+), Mn(2+), and Li(+) improved enzyme activity. 3-Cyanopyridine (50 mM) was hydrolyzed into nicotinic acid within 30 min, whereas only <5% of nicotinamide was detected. The results show that this fungal nitrilase is a promising candidate for commercial application in nicotinic acid production.


Subject(s)
Aminohydrolases/metabolism , Gibberella/enzymology , Gibberella/isolation & purification , Cluster Analysis , DNA, Fungal/chemistry , DNA, Fungal/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , DNA, Ribosomal Spacer/chemistry , DNA, Ribosomal Spacer/genetics , Enzyme Activators/analysis , Enzyme Inhibitors/analysis , Genes, rRNA , Gibberella/classification , Gibberella/genetics , Hydrogen-Ion Concentration , Molecular Sequence Data , Phylogeny , RNA, Fungal/genetics , RNA, Ribosomal, 18S/genetics , Sequence Analysis, DNA , Substrate Specificity , Temperature
14.
Acta Physiol (Oxf) ; 207(1): 166-72, 2013 Jan.
Article in English | MEDLINE | ID: mdl-22846709

ABSTRACT

AIM: Salusin-ß is a regulatory peptide that exerts negative inotropic effect on ventricular muscle, but its electrophysiological effects on ventricular myocytes are still unknown. METHODS: Action potential and channel currents such as sodium current (I(N) (a) ), transient outward potassium current (I(to) ), steady-state potassium current (I(sus) ), sodium-calcium exchange current (I(N) (aCa) ) and inward rectifier potassium current (I(K) (1) ) were measured in ventricular myocytes isolated from 12 to 16 weeks rats by whole-cell voltage-clamp techniques. RESULTS: Salusin-ß dose-dependently shortened the duration of action potential in rat ventricular myocytes. Furthermore, salusin-ß significantly inhibited I(N) (aCa) and increased I(to) , but did not affect I(N) (a) , I(sus) and I(K) (1) . CONCLUSION: These results suggest that the effect of salusin-ß on action potential may be partly attributed to a decrease in inward currents and an increase in outward currents.


Subject(s)
Action Potentials/physiology , Intercellular Signaling Peptides and Proteins/physiology , Myocardial Contraction/physiology , Myocytes, Cardiac/physiology , Action Potentials/drug effects , Animals , Cells, Cultured , Heart Ventricles/cytology , Intercellular Signaling Peptides and Proteins/pharmacology , Membrane Potentials/drug effects , Membrane Potentials/physiology , Myocardial Contraction/drug effects , Myocytes, Cardiac/cytology , Myocytes, Cardiac/drug effects , Patch-Clamp Techniques , Potassium Channels/physiology , Potassium Channels, Inwardly Rectifying/physiology , Rats , Sodium Channels/physiology , Sodium-Calcium Exchanger/physiology
15.
PLoS One ; 7(11): e50622, 2012.
Article in English | MEDLINE | ID: mdl-23226336

ABSTRACT

BACKGROUND: Nitrilase is an important member of the nitrilase superfamiliy. It has attracted substantial interest from academia and industry for its function of converting nitriles directly into the corresponding carboxylic acids in recent years. Thus nitrilase has played a crucial role in production of commercial carboxylic acids in chemical industry and detoxification of nitrile-contaminated wastes. However, conventional studies mainly focused on the bacterial nitrilase and the potential of fungal nitrilase has been far from being fully explored. Research on fungal nitrilase gene expression will advance our understanding for its biological function of fungal nitrilase in nitrile hydrolysis. METHODOLOGY/PRINCIPAL FINDINGS: A fungal nitrilase gene from Gibberella intermedia was cloned through reverse transcription-PCR. The open reading frame consisted of 963 bp and potentially encoded a protein of 320 amino acid residues with a theoretical molecular mass of 35.94 kDa. Furthermore, the catalytic triad (Glu-45, Lys-127, and Cys-162) was proposed and confirmed by site-directed mutagenesis. The encoding gene was expressed in Escherichia coli Rosetta-gami (DE3) and the recombinant protein with His(6)-tag was purified to electrophoretic homogeneity. The purified enzyme exhibited optimal activity at 45°C and pH 7.8. This nitrilase was specific towards aliphatic and aromatic nitriles. The kinetic parameters V(max) and K(m) for 3-cyanopyridine were determined to be 0.81 µmol/min·mg and 12.11 mM through Hanes-Woolf plot, respectively. 3-Cyanopyridine (100 mM) could be thoroughly hydrolyzed into nicotinic acid within 10 min using the recombinant strain with the release of about 3% nicotinamide and no substrate was detected. CONCLUSIONS/SIGNIFICANCE: In the present study, a fungal nitrilase was cloned from the cDNA sequence of G. intermedia and successfully expressed in E. coli Rosetta-gami (DE3). The recombinant strain displayed good 3-cyanopyridine degradation efficiency and wide substrate spectrum. This fungal nitrilase might be a potential candidate for industrial applications in carboxylic acids production.


Subject(s)
Aminohydrolases/genetics , Aminohydrolases/metabolism , Gibberella/enzymology , Histidine , Amino Acid Sequence , Aminohydrolases/chemistry , Aminohydrolases/isolation & purification , Cloning, Molecular , Gene Expression , Gibberella/genetics , Hydrolysis , Kinetics , Molecular Sequence Data , Nitriles/metabolism
16.
Appl Microbiol Biotechnol ; 92(2): 371-9, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21870045

ABSTRACT

In this study, alteration in morphology of submergedly cultured Antrodia camphorata ATCC 200183 including arthroconidia, mycelia, external and internal structures of pellets was investigated. Two optimization models namely response surface methodology (RSM) and artificial neural network (ANN) were built to optimize the inoculum size and medium components for intracellular triterpenoid production from A. camphorata. Root mean squares error, R (2), and standard error of prediction given by ANN model were 0.31%, 0.99%, and 0.63%, respectively, while RSM model gave 1.02%, 0.98%, and 2.08%, which indicated that fitness and prediction accuracy of ANN model was higher when compared to RSM model. Furthermore, using genetic algorithm (GA), the input space of ANN model was optimized, and maximum triterpenoid production of 62.84 mg l(-1) was obtained at the GA-optimized concentrations of arthroconidia (1.78 × 105 ml(-1)) and medium components (glucose, 25.25 g l(-1); peptone, 4.48 g l(-1); and soybean flour, 2.74 g l(-1)). The triterpenoid production experimentally obtained using the ANN-GA designed medium was 64.79 ± 2.32 mg l(-1) which was in agreement with the predicted value. The same optimization process may be used to optimize many environmental and genetic factors such as temperature and agitation that can also affect the triterpenoid production from A. camphorata and to improve the production of bioactive metabolites from potent medicinal fungi by changing the fermentation parameters.


Subject(s)
Antrodia/metabolism , Artificial Intelligence , Culture Media/chemistry , Fermentation , Triterpenes/metabolism , Antrodia/chemistry , Antrodia/genetics , Culture Media/metabolism , Neural Networks, Computer
17.
Appl Biochem Biotechnol ; 165(3-4): 963-77, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21720838

ABSTRACT

Microbial transformation of glycinonitrile into glycine by nitrile hydrolase is of considerable interest to green chemistry. A novel fungus with high nitrile hydrolase was newly isolated from soil samples and identified as Fusarium oxysporum H3 through 18S ribosomal DNA, 28S ribosomal DNA, and the internal transcribed spacer sequence analysis, together with morphology characteristics. After primary optimization of culture conditions including pH, temperature, carbon/nitrogen sources, inducers, and metal ions, the enzyme activity was greatly increased from 326 to 4,313 U/L. The preferred carbon/nitrogen sources, inducer, and metal ions were glucose and yeast extract, caprolactam, and Cu(2+), Mn(2+), and Fe(2+), respectively. The maximum enzyme formation was obtained when F. oxysporum H3 was cultivated at 30 °C for 54 h with the initial pH of 7.2. There is scanty report about the optimization of nitrile hydrolase production from nitrile-converting fungus.


Subject(s)
Aminoacetonitrile/metabolism , DNA, Ribosomal/analysis , Fungal Proteins/metabolism , Fusarium/enzymology , Hydrolases/metabolism , Industrial Microbiology/methods , Soil Microbiology , Caprolactam/metabolism , Carbon/metabolism , Culture Media , Fusarium/genetics , Fusarium/isolation & purification , Glucose/metabolism , Glycine/biosynthesis , Hydrogen-Ion Concentration , Hydrolysis , Metals, Heavy/metabolism , Nitriles/metabolism , Nitrogen/metabolism , Phylogeny , Temperature
18.
Zhong Yao Cai ; 34(11): 1722-5, 2011 Nov.
Article in Chinese | MEDLINE | ID: mdl-22506397

ABSTRACT

OBJECTIVE: To analyze the volatile compounds of Antrodia camphorata in solid-state and submerged cultures. METHODS: A headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry(GC-MS) were used to evaluate the profile of the volatile compounds. RESULTS: 49 volatile compounds were identified in A. camphorata mycelia in submerged culture, while 43 volatile compounds were identified in mycelia in solid-state culture. 1-octen-3-ol, 3-octanone, 1-octen-3-ylacetate, acetic acid octyl ester and ethanol were the main volatile compounds in A. camphorata mycelia in submerged culture, while 1-octen-3-ol, 3-octanone, 3-methyl-butyraldenhyde, gamma-podecalactone and methyl 2-furozte were the most potent key volatile compounds in mycelia in solid-state culture. CONCLUSION: The volatile compounds in the mycelia of A. camphorata in solid-state and submerged cultures are similar but their relative contents are different.


Subject(s)
Antrodia/chemistry , Mycelium/chemistry , Volatile Organic Compounds/analysis , Acetates/analysis , Culture Techniques/methods , Ethanol/analysis , Gas Chromatography-Mass Spectrometry/methods , Ketones/analysis , Octanols/analysis , Solid Phase Microextraction/methods
19.
Peptides ; 31(6): 1146-9, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20307603

ABSTRACT

Salusin-beta is a new regulatory peptide relevant to the cardiovascular system and exerts negative inotropic effect on ventricular muscle. The purpose of the present study was to determine whether salusin-beta can inhibit cardiac L-type calcium channel current (I(Ca,L)). Using whole-cell voltage-clamp techniques, I(Ca,L) was measured in ventricular myocytes isolated from 12 to 16 weeks rats. Salusin-beta dose-dependently and reversibly reduced the magnitude of I(Ca,L) in rat ventricular myocytes. Neither threshold potential nor the peak potential of current-voltage relationship was affected. Salusin-beta increased the rate of I(Ca,L) inactivation without altering its gating properties. These results suggest salusin-beta inhibited I(Ca,L) by increasing the rate of I(Ca,L) inactivation and the inhibition of L-type Ca(2+) channels induced by salusin-beta may contribute to its negative inotropic effect on ventricular muscle.


Subject(s)
Calcium Channels, L-Type/drug effects , Intercellular Signaling Peptides and Proteins/pharmacology , Myocytes, Cardiac/drug effects , Animals , Calcium Channels, L-Type/physiology , Male , Patch-Clamp Techniques , Rats , Rats, Sprague-Dawley
20.
Peptides ; 30(9): 1742-5, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19560502

ABSTRACT

Obestatin is a recently discovered 23-amino acid peptide encoded by the same gene that encodes ghrelin. It has been reported that there is a significant negative correlation between the plasma ghrelin concentration and systemic blood pressure in patients with pregnancy-induced hypertension. We investigated the plasma concentration of obestatin in 18 non-pregnant women, 18 normal pregnant women, and 15 patients with pregnancy-induced hypertension. The plasma concentrations of obestatin in these 3 groups of women were 63.4+/-9.5pg/ml, 38.1+/-6.3pg/ml, and 46.0+/-9.3pg/ml, respectively. In non-pregnant women, there was no correlation between the plasma obestatin concentration and the mean arterial pressure. However, there was a positive correlation between the plasma obestatin concentration and the mean arterial pressure in normal pregnant women and pregnant women with pregnancy-induced hypertension. These results suggest that obestatin may have some potential role in the regulation of blood pressure in normal pregnant women and women with pregnancy-induced hypertension.


Subject(s)
Blood Pressure/physiology , Peptide Hormones/blood , Pregnancy Trimester, Third/blood , Pregnancy Trimester, Third/physiology , Adult , Female , Fetal Blood/metabolism , Ghrelin , Humans , Hypertension, Pregnancy-Induced/blood , Pregnancy , Young Adult
SELECTION OF CITATIONS
SEARCH DETAIL
...