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1.
《Gene》1998,208(2):177-182
The Eco29kI restriction-modification system (RMS2) has been found to be localized on the plasmid pECO29 occurring naturally in the Escherichia coli strain 29k (Pertzev, A.V., Ruban, N.M., Zakharova, M.V., Beletskaya, I.V., Petrov, S.I., Kravetz, A.N., Solonin, A.S., 1992. Eco29kI, a novel plasmid encoded restriction endonuclease from Escherichia coli. Nucleic Acids Res. 20, 1991). The genes coding for this RMS2, a SacII isoschizomer recognizing the sequence CCGCGG have been cloned in Escherichia coli K802 and sequenced. The DNA sequence predicts the restriction endonuclease (ENase) of 214 amino acids (aa) (24 556 Da) and the DNA-methyltransferase (MTase) of 382 aa (43 007 Da) where the genes are separated by 2 bp and arranged in tandem with eco29kIR preceding eco29kIM. The recombinant plasmid with eco29kIR produces a protein of expected size. ṀEco29kI contains all the conserved aa sequence motifs characteristic of m5C-MTases. Remarkably, its variable region exhibits a significant similarity to the part of the specific target-recognition domain (TRD) from ṀBssHII—multispecific m5C-MTase (Schumann, J.J., Walter, J., Willert, J., Wild, C., Koch D., Trautner, T.A., 1996. ṀBssHII: a multispecific cytosine-C5-DNA-methyltransferase with unusual target recognizing properties. J. Mol. Biol. 257, 949–959), which recognizes five different sites on DNA (HaeII, MluI, Cfr10I, SacII and BssHII), and the comparison of the nt sequences of its variable regions allowed us to determine the putative TRD of ṀEco29kI.  相似文献   

2.
《Gene》1997,195(2):207-216
With lysogeny by bacteriophage SfV, Shigella flexneri serotype Y is converted to serotype 5a. The glucosyl transferase gene (gtr) from bacteriophage SfV of S. flexneri, involved in serotype-specific conversion, was cloned and characterized. The DNA sequence of a 3.7 kb EcoRI–BamHI fragment of bacteriophage SfV which includes the gtr gene was determined. This gene, encoding a polypeptide of 417 aa with 47.67 kDa molecular mass, caused partial serotype conversion of S. flexneri from serotype Y to type V antigen as demonstrated by Western blotting and the sensitivity of the hybrid strain to phage Sf6. The deduced protein of the partially sequenced open reading frame upstream of the gtr showed similarity to various glycosyl transferases of other bacteria. Orf3, separated from the gtr by a non-coding region and transcribed convergently, codes for a 167 aa (18.8 kDa) protein found to have homology with tail fibre genes of phage lambda and P2.  相似文献   

3.
Chemical and genetic diversity of Teucrium polium L. subsp. polium from western Algeria and T. polium L. subsp. capitatum from Corsica were investigated. Diversity within and among the two populations of subspecies was assessed according to the chemical composition of their essential oils and the genetic diversity. Chemical analysis was performed using a combination of capillary GC-RI and GC/MS after fractionation using column chromatography. Genetic structures were mapped using three polymorphic genetic markers: two chloroplast markers (RPL32-TRNL and TRNL-F) and ribosomal nuclear markers (ITS region). The statistical analysis showed that both subspecies were clearly distinguished by these chemical and genetic markers. The oil chemical compositions differed qualitatively and quantitatively between the subspecies. Both collective oils were dominated by hydrocarbon compounds however the Algerian sample oils exhibited higher amounts of hydrocarbon sesquiterpenes than those of Corsica (31.2 g/100 g vs. 4.4 g/100 g) while the latter displayed higher amounts of hydrocarbon monoterpenes than the first (59.3 g/100 g vs. 34.3 g/100 g). Neighbor-joining, Maximum likelihood and Bayesian trees constructed from chloroplast markers and nuclear ITS region sequences showed the existence of two groups associated with taxonomic and chemical characteristics. The study indicated that variation in the essential oil composition within subspecies depends on genetic background. The samples of subsp. capitatum from Corsica are a homogeneous group, in contrast to samples of subsp. polium from Algeria which were clustered in two groups. Chemical and genetic diversity of Algerian populations could be explained by geographical isolation of the populations. In addition, the morphological polymorphism observed throughout the colour of flowers could be explained by environmental parameters as well as the soil pH.  相似文献   

4.
In pathological conditions, the balance between reactive oxygen species (ROS) and antioxidants may shift toward a relative increase of ROS, resulting in oxidative stress. Conflicting data are available on antioxidant defenses in human failing heart and they are limited to the left ventricle. Thus, we aimed to investigate and compare the source of oxidant and antioxidant enzyme activities in the right (RV) and left (LV) ventricles of human failing hearts. We found a significant increase in superoxide production only by NADPH oxidase in both failing ventricles, more marked in RV. Despite unchanged mRNA or protein expression, catalase (CAT) and glutathione peroxidase (GPx) activities were increased, and their increases reflected the levels of Tyr phosphorylation of the respective enzyme. Manganese superoxide dismutase (Mn-SOD) activity appeared unchanged. The increase in NADPH oxidase-dependent superoxide production positively correlated with the activation of both CAT and GPx. However, the slope of the linear correlation (m) was steeper in LV than in RV for GPx (LV: m = 2.416; RV: m = 1.485) and CAT (LV: m = 1.007; RV: m = 0.354). Accordingly, malondialdehyde levels, an indirect index of oxidative stress, were significantly higher in the RV than LV. We conclude that in human failing RV and LV, oxidative stress is associated with activation of antioxidant enzyme activity. This activation is likely due to post-translational modifications and more evident in LV. Overall, these findings suggest a reduced protection of RV against oxidative stress and its potential contribution to the progression toward overt heart failure.  相似文献   

5.
Concerning the instability of ATP liposomes formulated to easily diffuse through the liver (size ~100 nm), this work targets the key parameters that influence the freeze-drying of a preparation that combines cholesterol, DOTAP and phosphatidylcholine (either natural soybean or egg (SPC or EPC) or hydrogenated (HSPC)). After freeze-drying blank liposomes, size increased significantly when initial lipid concentration was lowered from 20 to 5 mM (p = 0.0018). With low lipid concentration preparation (5 mM), SPC limited size increase (SI) more efficiently compared to EPC or HSPC. With SPC and EPC, sucrose showed better size results compared to trehalose (Lyoprotectant/Lipid ratio (w/w) avoiding any SI: ~5 and ~10 (for SPC), ~10 and ~15 (for EPC), for sucrose and trehalose, respectively), but the opposite was evidenced with HSPC liposomes where a Trehalose/Lipid ratio of 25 barely prevented SI. In addition, slow versus quick cooling rate led to limiting SI for HSPC liposomes (p = 0.0035). With sucrose or trehalose at both Lyoprotectant/Lipid ratios ensuring size stabilisation (10:1 and 15:1, respectively), ATP leakage ranged between 38.8 ± 7.9% and 58.2 ± 1.4%. In conclusion, this study emphasizes that using strict size maintenance as the primary objective does not result in drug complete retention inside the liposome core.  相似文献   

6.
A series of methoxylated chalcones with fluoro and trifluoromethyl derivatives were synthesized and investigated for their ability to inhibit human monoamine oxidase A and B. The chemical structures of the compounds have been characterized by means of their 1H NMR, 13C NMR, Mass spectroscopic datas and elemental analysis. The results demonstrate that these compounds are reversible and selective MAO-B inhibitors with a competitive mode of inhibition. The most potent compound (2E)-1-(4-methoxyphenyl)-3-[4-(trifluoromethyl)phenyl] prop-2-en-1-one showed the best activity and higher selectivity towards hMAO-B with Ki and SI values of 0.22 ± 0.01 μM and 0.05 comparable to that standard drug, Selegiline Ki and SI values were found as 0.33 ± 0.03 μM and 0.04, respectively. Molecular docking studies were carried out to further explain the in vitro results of the new compounds, and to identify the hypothetical binding mode for the compounds inside the inhibitor binding cavity of hMAO-B.  相似文献   

7.
The impact of a rectal spacer and an increased near maximum target dose in VMAT prostate SBRT is studied.For a group of 11 patients (35 Gy-in-five-fractions VMAT prostate SBRT) a set of 4 plans were generated, namely two VMAT plans, with D2%  37.5 Gy (Hom) and with D2%  40.2 Gy (Het), were created for each of two CT scans taken before (NoSpc) and after (Spc) transperineal spacer insertion. Consequently the methodology for parameter invariant TCP (tumor control probability) plan ranking was applied for comparison of the plans in terms of tumor control. NTCPs (normal tissue complication probabilities) were calculated for rectum and bladder using Lyman’s model.For all 11 patients the TCP plan ranking has shown that the Het plans would perform considerably better in TCP terms than the Hom ones. The plans without rectal spacer were ranked worse compared to those with rectal spacer except for one set of Hom plans. The calculated NTCPs for rectum produced by the Het plans were quite similar to the NTCPs of the Hom ones. The rectal NTCPs of the Hom Spc plans were always lower than the NTCPs of the Hom NoSpc plans. The NTCP values for bladder were extremely low in all cases.The use of rectal spacer leads in general to lower risk of rectal complications, as expected, and even to better tumor control. Plans with increased near maximum target dose (D2%  40.2 Gy) are expected to perform much better in terms of tumor control than those with D2%  37.5 Gy.  相似文献   

8.
《Small Ruminant Research》2010,91(1-3):161-164
Progesterone is essential for maintaining pregnancy in goats, and embryonic losses may be a consequence of the reduction in circulating progesterone levels close to the time of implantation. Some evidence exists regarding social dominance affecting the plasma progesterone levels in several species—where dominant females conceive earlier. The objective of this research was to determine whether serum progesterone levels differ in goats of different social status. A behavioural study was conducted for 10 days in a herd of 57 does and an individual success index (SI) was calculated according to the result of aggressive interactions. Goats were classified as high (SI: 1–0.67), medium (SI: 0.66–0.34) and low-ranking (SI: 0.33–0.0). Ovulation was synchronized using two injections of prostaglandin 11 days apart, and the plasma progesterone levels determined daily for a period of 20 days. The area under the plasma progesterone curve during the entire study was greater in the high than in the medium and low-ranking does (96.2 ± 5.8, 79.5 ± 5.3 and 81.3 ± 5.3 ng/ml, respectively, P < 0.05). During days 11–17 following prostaglandin synchronization, the plasma progesterone levels were higher in the high-ranking (P < 0.05), compared to the low-ranking does. Plasma progesterone levels were significantly correlated with SI at days 14 and 15 (r = 0.26; P < 0.05). Results suggest a relationship between social ranking of goats and the plasma progesterone production from the corpus luteum and other possible sources.  相似文献   

9.
10.
The 13C/12C ratios of leaves of the conifer morphotype Frenelopsis were measured to decipher the influences of water and salt stress on stomatal density (SD), epidermal cell density (ECD) and stomatal index (SI). Three morphospecies were analyzed: F. ugnaensis from freshwater fluvio-lacustrine deposits (Upper Barremian), F. turolensis and alata from coastal deposits (Lower-Middle Albian and Upper Albian respectively). The cuticle δ13C values show a large variation from ? 28‰ to ? 21‰. Comparison with previously published marine carbonate δ13C records indicate that the difference in cuticle δ13C between the different deposits are mainly due to difference in CO2-plant isotope fractionation rather than to change in isotopic composition of inorganic carbon in the atmosphere and ocean. The less negative δ13C and wide range in δ13C of F. turolensis and alata (? 27.5 to ? 21‰), compared to F. ugnaensis, (? 28 to ? 25‰) are interpreted as a result of salt and/or water stress. The data as a whole yield a good relationship between the 13C/12C ratio and SD (r = 0.67, n = 42, p < 0.001), SI (r = 0.53, n = 41, p < 0.001), hence suggesting that the differences in SD and SI between the three morphospecies are related to freshwater/saline environment. Looking at single morphospecies, the SD of F. ugnaensis decreases with increasing δ13C value (r = ? 0.57, n = 15, p = 0.026) as well as a decrease of SI (r = ? 0.62, n = 15, p = 0.013), possibly reflecting warmer and drier conditions. Average SI of F. alata does not significantly change with δ13C and inferred soil salinity in contrast to SD (p < 0.01).  相似文献   

11.
Current treatment for hepatitis C is barely satisfactory, there is an urgent need to develop novel agents for combating hepatitis C virus infection. This study discovered a new class of thieno[2,3-b]pyridine derivatives as HCV inhibitors. First, a hit compound characterized by a thienopyridine core was identified in a cell-based screening of our privileged small molecule library. And then, structure activity relationship study of the hit compound led to the discovery of several potent compounds without obvious cytotoxicity in vitro (12c, EC50 = 3.3 μM, SI >30.3, 12b, EC50 = 3.5 μM, SI >28.6, 10l, EC50 = 3.9 μM, SI >25.6, 12o, EC50 = 4.5 μM, SI >22.2, respectively). Although the mechanism of them had not been clearly elucidated, our preliminary optimization of this class of compounds had provided us a start point to develop new anti-HCV agents.  相似文献   

12.
Botulinum neurotoxins, responsible for the neuroparalytic syndrome botulism, are the deadliest of known biological toxins. The work described in this study was based on a three-zone pharmacophore model for botulinum neurotoxin serotype A light chain inhibition. Specifically, the pharmacophore defined a separation between the overlaps of several different, non-zinc(II)-coordinating small molecule chemotypes, enabling the design and synthesis of a new structural hybrid possessing a Ki = 600 nM (±100 nM).  相似文献   

13.
Hypericum perforatum L. (St. John’s wort, SJW) is a traditional medicinal plant with a long history of pharmaceutical application. Today, besides formulations based on hydro-alcoholic extracts, lipophilic SJW preparations are widely used in phytomedicine e.g. for the symptomatic treatment of minor inflammations of the skin and subcutaneous tissue disorders. In the present study, SJW extracts were obtained with twelve different fatty oils according to a standardized protocol. In order to evaluate and compare the resulting macerates, a set of analytical parameters was chosen: the phytochemical profile of SJW oil extracts comprising acylphloroglucinol derivatives, flavonols, biflavones and naphthodianthrones was assessed using UHPLC-DAD and UV/VIS-spectroscopic methods Furthermore, overall appearance was monitored applying the CIE L*a*b* color system. Depending on the type of oil applied for extraction, a*-values, which are a measure of red color hues, ranged from 50.3 ± 0.2 (almond oil) to −3.8 ± 0.3 (macadamia nut oil). Considering total hypericin contents, extraction with almond oil also brought about highest amounts (5.5 ± 0.21 mg/100 g). In contrast, preparations with macadamia nut oil resulted in highest contents of hyperforin and adhyperforin, whereas a semi-synthetic extractant composed of medium-chain triglycerides (MCT) revealed highest yields of flavonoids, i.e. quercetin, kaempferol and I3, II8-biapigenin. Since the phytochemical composition and the corresponding specific characteristics of SJW oil preparations markedly differed, tailor-made SJW extracts may be prepared by systematic adaption of the manufacturing conditions, including the choice of a specific fatty oil. Additionally, SJW oil extracts were stored under different temperature (5 °C, 20 °C, 50 °C) and light (artificial light, darkness) regimes over a period of six months to evaluate their respective storage stability. For maximizing the retention of the investigated plant secondary metabolites in the corresponding oil extracts, cold storage (5 °C) under the exclusion of light is recommended.  相似文献   

14.
The study aimed to map instantaneous centers of rotation (ICRs) of lumbar motion segments during a functional lifting task and examine differences across segments and variations caused by magnitude of weight lifted. Eleven healthy participants lifted loads of three different magnitudes (4.5, 9, and 13.5 kg) from a trunk-flexed (~75°) to an upright position, while being imaged by a dynamic stereo X-ray (DSX) system. Tracked lumbar vertebral (L2-S1) motion data were processed into highly accurate 6DOF intervertebral (L2L3, L3L4, L4L5, L5S1) kinematics. ICRs were computed using the finite helical axis method. Effects of segment level and load magnitude on the anterior-posterior (AP) and superior-inferior (SI) ICR migration ranges were assessed with a mixed-effects model. Further, ICRs were averaged to a single center of rotation (COR) to assess segment-specific differences in COR AP- and SI-coordinates. The AP range was found to be significantly larger for L2L3 compared to L3L4 (p=0.02), L4L5 and L5S1 (p<0.001). Average ICR SI location was relatively higher – near the superior endplate of the inferior vertebra – for L4L5 and L5SI compared to L2L3 and L3L4 (p≤0.001) – located between the mid-transverse plane and superior endplate of the inferior vertebra – but differences were not significant amongst themselves (p>0.9). Load magnitude had a significant effect only on the SI component of ICR migration range (13.5 kg>9 kg and 4.5 kg; p=0.049 and 0.017 respectively). The reported segment-specific ICR data exemplify improved input parameters for lumbar spine biomechanical models and design of disc replacements, and base-line references for potential diagnostic applications.  相似文献   

15.
Gulf Menhaden (Brevoortia patronus) are a species of commercial and ecological importance in the northern Gulf of Mexico, provisioning the second largest fishery by weight, in the United States, and providing critical ecosystem services in the coastal region. The recruitment and productivity dynamics of the stock are influenced by a suite of environmental factors but an understanding of the factors that determine individual variation in oil content (an indicator of an individual’s commercial value to the fishery and its dietary value to predators) has not been well described. In this work I describe the temporal dynamics of oil content and determine the demographic characteristics that provide predictive power to describe annual contrasts. I relate the predicted patterns in oil yield to a suite of seasonal environmental data series including: the magnitude of spring Mississippi River discharge, spring wind vectors, and the preceding winter El Nino conditions. Two uncorrelated (r = 0.06, p = 0.81) population-level predictor variables were identified that have explanatory power to describe temporal patterns in oil content (L kg−1); a weight-at-length power function parameter (a) and the von Bertalanffy asymptotic fork length (L, mm FL): L kg−1 =  0.158  0.026*a  0.00163*L (p < 0.05, R2 = 0.42). Analysis of the impacts of environmental variables on the oil content of Gulf Menhaden was evaluated comprehensively in a Bayesian framework by transforming the observed oil content information from two sources to a common scale. Parameters relating oil content to spring Mississippi River discharge and the preceding winter (December–February) El Nino Southern Oscillation index resulted in sample distributions from the posterior where zero was outside the 95% credible interval. This work contributes to the understanding of Gulf Menhaden as a prey species in the Gulf of Mexico and indicates that the value of the species to both the fishery and predators exhibits relatively large inter-annual variability controlled, in part, by seasonal environmental conditions.  相似文献   

16.
We investigated the effect of lipopolysaccharide (LPS) chemotypes differing in their carbohydrate chain length on phagocytosis of serum-opsonized zymosan (OZ) particles and related functions of human polymorphonuclear leukocyte (PMNL, neutrophils). LPS from deep core mutant (Re), complete core (Ra) and smooth (S) phenotypes of Salmonella typhimurium was studied. Priming of neutrophils with various LPSs caused prominent enhancement of OZ phagocytosis, superoxide production and leukotriene (LT) synthesis in neutrophils, with LPS effects increasing as Re < S < Ra. The LPS forms were less potent to activate OZ uptake in the presence of MK-886, 5-lipoxygenase activating protein inhibitor, suggesting the regulatory function of 5-lipoxygenase (5-LO)-derived LTs. Direct measurement of nitrite release from OZ-stimulated neutrophils revealed that the effects of LPS on NO synthesis increased in the range of Ra < S < Re. Nitric oxide synthase (NOS) inhibitor l-NAME increased phagocytosis, LT and superoxide formation by neutrophils, and abolished the difference in the action of the LPSs forms. Further mechanistic studies revealed that NO modulates cellular 5-LO activity in a guanylyl cyclase and protein kinase G dependent manner, as well as interplay between NO and superoxide, and peroxynitrite generation contribute to distinct effects of LPS chemotypes on phagocytosis and LT synthesis in human neutrophils. Our investigation of the three LPS species demonstrates that the LPS polysaccharide core is mostly essential for the PMNL activation and is able to suppress lipid A-induced increase in NOS activity in phagocyting cells by triggering specific signaling cascades.  相似文献   

17.
《Phytomedicine》2014,21(11):1298-1302
ObjectiveThe essential oil from the leaves of Ocimum kilimandscharicum (EOOK), collected in Dourados-MS, was investigated for anticancer, anti-inflammatory and antioxidant activity and chemical composition.Materials and methodsThe essential oil was extracted by hydrodistillation, and the chemical composition was performed by gas chromatography–mass spectrometry. The essential oil was evaluated for free radical-scavenging activity using the DPPH assay and was tested in an anticancer assay against ten human cancer cell lines. The response parameter (GI50) was calculated for the cell lines tested. The anti-inflammatory activity was evaluated using carrageenan-induced pleurisy in mice.ResultsThe chemical composition showed 45 components with a predominance of monoterpenes, such as camphor (51.81%), 1,8 cineole (20.13%) and limonene (11.23%). The EOOK exhibited potent free radical-scavenging activity by the DPPH assay with a GI50 of 8.31 μg/ml. The major constituents, pure camphor (IC50 = 12.56 μg/ml) and mixture of the limonene: 1, 8 cineole (IC50 = 23.25 μg/ml) displayed a potent activity. The oral administration of EOOK (at 30 and 100 mg kg−1), as well as the pure camphor or a mixture of 1,8 cineole with limonene, significantly inhibited the carrageenan (Cg) induced pleurisy, reducing the migration of total leukocytes in mice by 82 ± 4% (30 mg kg−1 of EOOK), 95 ± 4% (100 mg kg−1 of EOOK), 83 ± 9% (camphor) and 80 ± 5% (mixture of 1,8 cineole:limonene 1:1). In vitro cytotoxicity screening against a human ovarian cancer cell line displayed high selectivity and potent anticancer activity with GI50 = 31.90 mg ml−1. This work describes the anti-inflammatory, anticancer and antioxidant effects of EOOK for the first time.ConclusionsThe essential oil exhibited marked anti-inflammatory, antioxidant and anticancer effects, an effect that can be attributed the presence of majorital compounds, and the response profiles from chemical composition differed from other oils collected in different locales.  相似文献   

18.
Exopolysaccharide produced by a marine Enterobacter cloaceae (designated as EPS 71a) emulsified hexane, benzene, xylene, kerosene, paraffin oil, cottonseed oil, coconut oil, jojoba oil, castor oil, groundnut oil and sunflower oil. However, it could form stable emulsions with groundnut oil and hexane at optimal concentration of 1 mg ml−1. Further increase in concentration of EPS 71a did not show noteworthy increase in emulsification indices. Emulsions with groundnut oil and hexane were stable up to 10 days between pH 2 and 10 and in the presence of sodium chloride in the range of 5–50 mg ml−1 at 35–37 °C. EPS 71a formed stable emulsion with xylene as compared to commercial gums such as arabic, tragacanth, karaya and xanthan.  相似文献   

19.
A novel series of triazole integrated phenyl heteroterpenoids have been synthesized and screened for their in vitro activity against intracellular amastigote form of Leishmania donovani. Among all tested compounds, compound 3a was found to be the most active with IC50 6.4 μM and better selectivity index (SI) 18 as compared to reference drugs, miltefosine and miconazole. When evaluated in vivo in L. donovani/hamster model, 3a has exhibited 79 ± 11% inhibition of parasite multiplication at 50 mg kg?1 × 5 days on day 7 post treatment.  相似文献   

20.
Gemini surfactants from the homologous series of alkane-α,ω-diyl-bis(dodecyldimethylammonium bromide) (CnCS12, number of spacer carbons n = 2  12) and dioleoylphosphatidylethanolamine (DOPE) were used for cationic liposome (CL) preparation. CLs condense highly polymerized DNA creating complexes. Small-angle X-ray diffraction identified them as condensed lamellar phase LαC in the studied range of molar ratios CnGS12/DOPE in the temperature range 20  60 °C. The DNA–DNA distance (dDNA) is studied in dependence to CnGS12 spacer length and membrane surface charge density. The high membrane surface charge densities (CnGS12/DOPE = 0.35 and 0.4 mol/mol) lead to the linear dependence of dDNA vs. n correlating with the interfacial area of the CnGS12 molecule.  相似文献   

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