Comparison of different methods for protein determination inAspergillus niger mycelium

Summary Protein contents were determined in submerged as well as in surface-grown citric acid producingAspergillus niger mycelia. Various methods (Kjeldahl, Biuret, Lowry and Coomassie Blue) for protein determination were compared. The Biuret method seemed to be more suitable than the others for true protein determination in mycelia. The Lowry method gave lower results in all cases. The Coomassie Blue method did not prove suitable for the material used.     

The measurement of actin concentration in solution: a comparison of methods

Intrinsic optical density, Folin, and Biuret color development have been carefully studied as methods of determining actin concentration in solution. It appears that the Lowry (Folin) method is the most sensitive and reliable method as standardized by Kjeldahl analysis. Intrinsic optical density is also found to be a reliable method and the extinction coefficients of F and G actin at 280 and 290 nm are determined. The Biuret reaction is found to be the least reliable of the three methods for determining the concentration of actin in solution.     

Concentration and composition of plasma proteins in wild mammals

Total protein concentrations from a variety of wild mammalian species were measured by the Lowry and Biuret techniques and compared with results obtained or derived by refractometry. Values obtained with the Lowry method were the highest and certain derived refractometry results, the lowest. It is suggested that the Biuret method provides the most acceptable values but that care should be exercised in using the correct standard. The percentage composition of plasma from 22 species of wild mammals is also presented and the observations discussed with relation to methodology and A/G ratio.     

黄热病病毒检测微阵列的研究制备

目的:制备黄热病病毒寡核苷酸检测微阵列.方法:根据黄热病病毒基因组序列,并应用生物信息学软件设计出寡核苷酸探针用于制备基因芯片,克隆于质粒上的黄热病病毒全长基因DNA经限制性显示技术扩增并标记,完成杂交后对芯片进行扫描和数据分析.结论:微阵列检测技术为检测黄热病病毒提供了一种早期、快速、可靠的方法,具有应用于临床检测的前景.     

4'-C-methyl-beta-D-ribofuranosyl purine and pyrimidine nucleosides revisited

In order to evaluate their antiviral properties, a series of 4'-C-methyl-beta-D-ribofuranosyl purine and pyrimidine nucleosides has been prepared. Unfortunately, none of these 4'-branched nucleosides showed any antiviral activity or cytotoxcity when tested against HIV, HBV, and Yellow Fever virus.     

Worldwide invasion of vector mosquitoes: present European distribution and challenges for Spain

An Asiatic mosquito species, Aedes albopictus, began to spread worldwide in the 1970s thanks to marine transport of tires and other goods, leading to colonization of many areas of the world. This species is a vector of major human diseases such as Dengue, Yellow Fever and the West Nile virus. In Europe, it was established in Albania and Italy and has been detected in other countries such as France; no records exist for Spain as yet. Colonization by Aedes albopictus is a major public health concern considering that the West Nile virus and several other viruses are known to circulate sporadically in the Mediterranean. Additionally, the parent species Aedes aegypti was the vector causing severe outbreaks of Dengue and Yellow Fever two centuries ago. Although Ae. aegypti was also introduced, it was eradicated from Spain. Both mosquitoes shared habitat types, diseases transmitted and many bionomic data. This article contains a review of the present Ae. albopictusdistribution range worldwide and discusses the likelihood of an establishment in Spain in view of climatological and geographical data. This revised version was published online in July 2006 with corrections to the Cover Date.     

High-level gene expression in Aedes albopictus cells using a baculovirus Hr3 enhancer and IE1 transactivator

Background  

Aedes aegypti is the key vector of both the Yellow Fever and Dengue Fever viruses throughout many parts of the world. Low and variable transgene expression levels due to position effect and position effect variegation are problematic to efforts to create transgenic laboratory strains refractory to these viruses. Transformation efficiencies are also less than optimal, likely due to failure to detect expression from all integrated transgenes and potentially due to limited expression of the transposase required for transgene integration.     

Mathematical model to assess the control of Aedes aegypti mosquitoes by the sterile insect technique

We propose a mathematical model to assess the effects of irradiated (or transgenic) male insects introduction in a previously infested region. The release of sterile male insects aims to displace gradually the natural (wild) insect from the habitat. We discuss the suitability of this release technique when applied to peri-domestically adapted Aedes aegypti mosquitoes which are transmissors of Yellow Fever and Dengue disease.     

测定胸腺肽含量几种定量方法的比较

本文用凯氏定氮法、双缩脲法及福林－酚法测定了胸腺肽含量,并加以比较,说明三种方法对其含量测定没有显著差异．在实践中可以根据不同条件选择适当的方法进行实验．     

用气相色谱法检测甲型肝炎减毒活疫苗中三氯甲烷含量

用气相色谱法检测甲型肝炎减毒活疫苗中的三氯甲烷含量。以正丙醇作内标,用峰高代替峰面积,采用HP INNOW_(ax)毛细管柱,程序升温方法,准确定量甲型肝炎减毒活疫苗中的三氯甲烷含量。结果显示,色谱法优于限量法,能准确定量,快速灵敏。色谱法可用于实际检测分析。     

In vitro potency assay for yellow fever vaccines: comparison of three vero cell lines sources.

Quality control of Yellow Fever vaccines performed by Control Authorities prior to marketing vaccines batches requires in vitro potency assays. The two currently available methods are the plaque formation assay and the cytopathic effect assay based on the use of porcine kidney PS cells or monkey kidney Vero cells. Among several sources of variation in virus titration, the cell systems are considered as important issues and Quality Assurance strongly recommends working with cell banks from certified suppliers. The aim of our study was to compare the behaviour and the sensitivity of three Vero cell sources obtained from ATCC, WHO and EP used at different passage levels in a plaque formation test. The conclusion of this work was that the yellow fever live attenuated virus titration, adapted in Vero cell lines appeared as a reliable method applicable for routine in vitro potency assay. The comparison of Vero cell lines, originated from three different sources, showed that they could be equally used as substrates by laboratories having the basic facility of cell culture, without influence on the final viral titre.     

4′- C -Methyl-β-D-ribofuranosyl Purine and Pyrimidine Nucleosides Revisited

Abstract

In order to evaluate their antiviral properties, a series of 4′-C-methyl-β-D-ribofuranosyl purine and pyrimidine nucleosides has been prepared. Unfortunately, none of these 4′-branched nucleosides showed any antiviral activity or cytotoxcity when tested against HIV, HBV, and Yellow Fever virus.     

Larvicidal effects of fungal Meroterpenoids in the control of Aedes aegypti L., the main vector of dengue and Yellow fever

The mosquito Aedes aegypti is an increasing problem of public health, being the vector responsible for dengue and Yellow Fever in tropical and subtropical regions. The aim of this work was to determine the potential larvicidal activity of a series of meroterpenoids, compounds 1-7, previously obtained fungal secondary metabolites from Penicillium sp., against the third-instar larvae of A. aegypti. The lethal concentrations (LC(50) and LC(90)) of 1-7 were evaluated 24 h after exposure. Dehydroaustin (4) was the most active meroterpenoid in the series, with an LC(50) value of 2.9 ppm, making it an attractive natural insecticide.     

A critical reappraisal of Waddell's technique for ultraviolet spectrophotometric protein estimation

Waddell's method of estimating protein concentration by the difference between spectrophotometric absorptions (215-225 nm) has been reexamined. Over limited ranges of total protein, a linear relation was found for ten purified proteins; the narrowest range was between 5 and 25 micrograms/ml. Using published extinction coefficients at 280 nm for these ten proteins, protein concentration at 280 nm correlated closely with the 215 nm/225 nm difference measurements (mean difference of 2.6%). Waddell's method also accurately determined the total protein in a mixture of proteins with widely varying individual 280-nm extinction coefficients. Biuret estimates of total protein in plasma or serum gave poor correlation with measurements by Waddell's method. Within protein concentration limits, Waddell's method was linear, narrow, and more variable, both for individual proteins and for protein mixtures, than previously reported. Within these limits, the method is probably as accurate a measure of total protein as measurement by nitrogen analysis, with the advantage of being nondestructive.     

Mutation and docking studies on NS2b-NS3 complex from yellow fever virus towards drug discovery

Yellow fever virus is the causative agent of Yellow fever. The genome of the virus contains three structural and seven non-structural proteins. Of these seven nonstructural proteins, NS2B-NS3 protein complex has protease activity required for viral replication. Predicting the 3D structure of this complex and studying the interaction of residues at the recognized catalytic triad of the complex is an integral part to understand the virus replication mechanism. In the present study, the structure was determined for NS2B-NS3 complex by Homology modeling and modeled structure was validated for its stability. Mutation studies at the residues His94, Asp118 and Ser176 revealed that Asp118-His94 bond played an important role in the structural stability of NS2B-NS3 complex. This indicates site-directed mutagenesis, controlling YFV replication, as one mechanism to design vaccine strains. Docking studies of the bioactive compounds at the active site of NS2B-NS3 complex also indicated 4-hydroxypanduratin A as potential lead compound for drug development. The theoretical models will further pave way to experimentally verify our mutation and docking studies, thus taking a lead in pharmacogenomics and drug development.

Abbreviations

YFV - Yellow Fever Virus, WNV - West Nile Virus, H-bonds - hydrogen bonds, SNP - Single nucleotide polymorphism.     

Metabolic and febrile responses to typhoid vaccine in humans: effect of beta-adrenergic blockade.

Fever and activation of acute phase responses were induced in human volunteers by intramuscular injection of typhoid vaccine. Vaccine injection caused a rapid (within 1 h) and sustained rise in metabolic rate (peak response 16%, 6-8 h), followed by later increases in white blood cell count (3-4 h), skin temperature (4-5 h), oral temperature (5-6 h), heart rate (6-8 h), and plasma cortisol (5-8 h). A peak fever [1.2 +/- 0.2 degree C (SE) rise] was recorded 12 h after vaccine injection. The involvement of the sympathetic nervous system in the development of these responses was investigated by the oral administration of propranolol before (80 mg) and 3 h after (40 mg) vaccine injection. Propranolol prevented the increases in metabolic rate, heart rate, and skin temperature but did not inhibit the rise in oral temperature or white cell count after vaccine administration. These data indicate that the sympathetic nervous system is responsible for the rise in energy expenditure associated with fever in humans. However, the rise in body temperature can develop in the absence of this increase in metabolic rate possibly by changes in heat loss.     

Immunogenicity and tumor protective activity of B16 melanoma vaccines

The immunogenicity and tumor-protective activity of different vaccines were examined and compared with murine B16 melanoma. All vaccines were prepared from material shed into culture medium by B16 melanoma cells. Vaccine I was generated by concentrating the shed material. Vaccine II was partially purified by precipitating the shed material with 50% ammonium sulfate followed by sephadex G-200 column chromatography. Vaccine III was concentrated shed material that was treated with 0.5% NP-40 and then ultracentrifuged to remove transplantation antigens. Mice were immunized to equal protein concentrations of vaccines weekly for 5 weeks or to control buffer. Antibody, cellular, and tumor-protective immunity to melanoma was measured in all mice 2 weeks following the last immunization. All three vaccine preparations were immunogenic. Vaccine preparation I appeared to be the most immunogenic and the one that most consistently augmented tumor-protective immunity. Augmentation in tumor-protective immunity correlated better with increase in cellular than in humoral immunity to melanoma.     

A comparison of the indirect haemagglutination test with the toxin neutralization test for the estimation of diphtheria antitoxin in mouse sera

Serum samples from 42 groups of mice immunized for different immunization periods with various doses of Adsorbed Diphtheria-Tetanus Vaccine, Adsorbed Diphtheria-Tetanus and Pertussis Vaccine and a standard diphtheria toxoid were assayed for their diphtheria antitoxin content by indirect haemagglutination (IHA) and by toxin neutralization (TN) tests. A very good correlation of 0.91 was obtained between the results of the two methods. There was no statistically significant difference between the IHA and the TN titres obtained. Adsorption with sheep red cells and treatment of the sera with 2-mercaptoethanol had no effect on the IHA titres. The minimum level of antitoxin detectable by the IHA test was 0.00039 IU ml-1. IHA proved to be a sensitive, specific and reproducible method which can be used reliably for the assay of diphtheria antitoxin in mouse sera.