Physical and Chemical Properties of Cilia Isolated from Tetrahymena pyriformis*

SYNOPSIS. Cilia devoid of their basal granules and isolated from Tetrahymena pyriformis strain W, by the method of Watson et al. (66) have been characterized in terms of physical, chemical and immunologic properties. The general chemical composition of cilia, based on mean values from these determinations is 66.3% protein, 24.3%“lipid,” 3.7 or 4.8% carbohydrate, and 0.4% nucleic acid. The amino acid composition of cilia includes hydroxyproline and is not significantly different from the amino acid composition of whole cells. Chloroformmethanol extracts of ciliary material contain free amino acids as well as neutral lipids and phospholipids. The lipid composition of cilia also is not significantly different from that of whole cells. Pentose, hexose and small but significant amounts of adenine nucleotide and ribonucleic acid are present. Cilia are a greater stimulus for the production of antibody than an equivalent amount of cellular antigen, and the ciliary antigen is strain specific.     

Giemsa Stain for the Diagnosis of Bovine Babesiosis. II. Changes in Erythrocytes Infected with Babesia bigemina and B. argentina

SYNOPSIS. Cytoplasmic stippling, intensification of the cell margin, and alterations in color, which have been reported in erythrocytes parasitized by Plasmodium falciparum in man, have been seen also in bovine erythrocytes parasitized by either Babesia bigemina or B. argentina. These changes appear to be identical in the human and bovine infections.
Tests with each component of Giemsa stain in simple aqueous solutions alone and in various combinations with eosin, together with tests with Giemsa stains containing one azure component, showed that demonstration of the changes depends on the presence of azure A and eosin and on prolonged staining times at pH 7.2 to 7.4. Specific tests suggested that the changes represent catabolic by-products of the parasites.     

Isolation from Pasteurella multocida of a Lipopolysaccharide Antigen with Immunizing and Toxic Properties

A heat-stable, particulate, lipopolysaccharide-protein antigenic complex has been isolated from a virulent, encapsulated strain of Pasteurella multocida by extraction with cold, formalinized saline, and centrifugation at 105,000 x g. The original bacterial culture had been obtained from a bison that died of hemorrhagic septicemia, an infectious disease of cattle and buffalo. Injection of fractional milligram amounts of the antigen into mice, rabbits, and calves produced toxic reactions which frequently resulted in death of the host. The surviving animals demonstrated a high degree of immunity to challenge with live, virulent organisms. Two injections with 15 mug of the antigen produced a high degree of immunity in mice without the development of any signs of toxicity. The gross chemical composition and toxicity of the antigen were similar to those reported for endotoxins obtained by the Boivin or Westphal procedure. Although strong serological cross-reactions were obtained in Ouchterlony plates between the 105,000 x g antigens from the bison strain and an avian strain with antisera to these strains, these antisera agglutinated only the bacterial cells of the homologous strain. The active immunity obtained in mice by the injection of the 105,000 x g antigens of each strain was specific and could be correlated with the agglutination tests.     

Serologic Changes Associated with the Encystment of Axenically Grown Hartmannella culbertsoni*

SYNOPSIS. Serologic reactions elicited by sonically ruptured trophic and cystic forms of Hartmannella culbertsoni were studied. The antigens of trophic amoebae reacted with their homologous rabbit antiserum showing multiple precipitin lines which could not be seen when the reacting antigens were treated with trypsin prior to application on the Ouchterlony plates. Antigens of trophic amoeba did not react with antiserum against cysts. Cyst antigens reacted with their homologous antiserum only after trypsin treatment. Antigens prepared from trophozoites excysting from cysts reacted positively with the antiserum against antigens of trophic amoebae. Antigens of trophic as well as cystic forms fixed guinea pig complement in presence of their homologous antisera. With the trophic form, this property was abolished after trypsin treatment. Non-specific complement fixation mediated by cyst antigens was abolished by treatment with cellulase. Antiserum against trophic amoebae immobilized trophozoites and, in the presence of guinea pig complement, led to their lysis.     

Physical and Textural Properties of Mayonnaise Prepared Using Virgin Coconut Oil/Fish Oil Blend

Physical and textural properties of mayonnaise prepared using virgin coconut oil (VCO)/fish oil (FO) blends at different ratios were examined in comparison with that prepared using soybean oil (SO) as affected by storage time (30 days). At day 0, sample prepared with SO showed the highest L*, a*, and b* values among all the samples, whereas the lowest values were noticeable for VCO containing sample. At day 30 of storage, decreases in L*, and b* values of all mayonnaise samples were observed (p < 0.05). However, a* values were increased at day 30 of storage (p < 0.05). For texture analysis, highest firmness, consistency and cohesiveness were obtained for the sample containing SO. Increasing levels of FO in VCO/FO samples increased the firmness, consistency and cohesiveness. For all the samples, loss modulus (G″) values were lower than G′. After 30 days of storage, all samples demonstrated slight decreases in G′ and viscosity than freshly prepared mayonnaise (day 0). When the sample containing VCO/FO (90:10) blend was further characterized, slight difference was observed in microscopic structure and droplet size distribution before and after storage of 30 days. Increase in droplet size was noticeable because of coalescence after the storage. Overall, type of oil used for preparation of mayonnaise as well as storage time affected the physical properties including textural and rheological properties of mayonnaise.

     

Preparation and Properties of Mitochondria from Naegleria gruberi*

Whole cell respiration rates were measured polarographically for Naegleria gruberi during growth in agitated cultures. Log growth phase amebae consumed 80 ng atoms O/min/mg cell protein. At stationary phase, respiration rate decreased 4–fold. Intact mitochondria were isolated from N. gruberi and their oxidative and phosphorylative capacities were studied polarographically. As with the mammalian system, the mitochondria oxidized succinate and NAD-linked substrates, but unlike rat liver mitochondria, those from the protozoan rapidly oxidized citrate and NADH. The rates of substrate oxidation were ADP-dependent, with ADP:O ratios equalling ? 2.8 for NAD-linked substrates and ? 2.2 for succinate. The respiratory control ratios. 2 to 4 for 11 substrates, were dependent on Pi, Mg²⁺, and serum albumin. Potassium cyanide, azide, malonale, and rotenone inhibited electron transport the same way as that of the mammalian system: however, amytal inhibited both glutamate and succinate respiration. Pentachlorophenol, DNP, and bilirubin uncoupled oxidation from phosphorylation. Difference spectra of oxidized and dithionite-reduced mitochondria had distinct absorption bands of flavins and of c-, b-, and α-type cytochromes.     

Extraction and Preliminary Use for Diagnosis of Soluble Precipitating Antigen from Babesia bigemina*

Soluble antigen from bovine blood with a Babesia bigemina parasitemia of 5–6% formed 4 precipitin lines in gel diffusion tests with antiserum from infected cattle. Antigen was obtained from plasma and hemolysate by elution from DEAE cellulose columns and (NH₄)₂SO₄ precipitation as well as from washed parasite-erythrocyte stroma by sonication or freezethawing. About 5 ml of antigen suitable for diagnostic tests could be extracted from 200 ml of infected blood.     

Indirect Hemagglutination with Mycoplasma Antigens: Effects of pH on Antigen Sensitization of Tanned Fresh and Formalinized Sheep Erythrocytes

An investigation of the influence of different factors affecting the sensitivity of the indirect hemagglutination test has been performed with antigens of four mycoplasmas isolated from sheep or goat. Tanned erythrocytes of sheep, fresh and formalinized, were sensitized with the above antigens. It was demonstrated that, with formalinized erythrocytes, the sensitivity was increased by 50 to 100 times when the sensitization was done at a low pH level. The pH level was unimportant for sensitizing fresh erythrocytes. The greatest sensitivity of the indirect hemagglutination test was obtained with fresh rather than formalinized erythrocytes. Three different types of antigens were used, and the most suitable antigen was found to be the supernatant fluid from an ultrasonically treated centrifuged Mycoplasma suspension.     

Physicomechanical Properties of Naproxen-Loaded Microparticles Prepared from Eudragit L100

Microparticles of naproxen with Eudragit L100 and Aerosil were prepared by the emulsion solvent diffusion method in order to avoid local gastrointestinal irritation, one of the major side effects of nonsteroidal anti-inflammatory drugs after oral ingestion. The process of preparation involved the use of ethanol as good solvent, dichloromethane as a bridging liquid, water as poor solvent, Aerosil as anti-adhesion agent, and sodium dodecyl sulfate to aid in the dispersion of the drug and excipients into the poor solvent. The obtained microparticles were evaluated for micromeritic properties, yield, encapsulation efficiency, drug physical state, and drug release properties. The influence of formulation factors and preparation condition (polymer/naproxen ratio, Aerosil/polymer ratio, and the initial difference of temperature between the solvent and nonsolvent) on the properties of the microparticles were also examined. The resultant microparticles were finely spherical and uniform with high incorporation efficiency (>79%) and yield (>71%). The incorporation efficiency was enhanced with increasing the ratio of excipients to drug and the initial difference of temperature between the solvent and nonsolvent. The mean diameter of the microparticles was influenced by all of the manufacturing parameters. Studies carried out to characterize the micromeritic properties of formulations, such as flowability and packability, showed that microparticles were suitable for further pharmaceutical manipulation (e.g., capsule filling). Drug release studies of the microparticles confirmed the gastroresistance, and mathematical studies showed that the drug released followed a Hixon and Crowell kinetic. These microparticles represent a simple method for the preparation of drug-loaded enteric microparticles with desired micromeritic properties and gastroresistance release.     

北京鸭红细胞铜锌超氧化物歧化酶电荷异构体的分离和某些性质

等电聚焦表明,北京鸭红细胞铜锌超氧化物歧化酶由等电点分别为５．０,５．３,５．９,６．１和６．５的五个主要的活性组分（电荷异构体）构成,利用分析型聚丙烯酰胺凝胶等电聚焦电泳进行电荷异构体的制备级分离,采用三氯乙酸沉淀法快速确定蛋白条带的位置,电渗洗脱法回收蛋白,获得其中两个电荷异构体,对比研究结果表明电荷异构体的活性,氨基酸组成,二级结构等性质无明显差异。     

Effects of Melatonin on Carbonic Anhydrase from Human Erythrocytes In Vitro and from Rat Erythrocytes In Vivo

The in vitro effects of melatonin (N-acetyl-5-methoxytryptamine) on human carbonic anhydrase isozymes (HCA-I and HCA-II) from human erythrocytes and in vivo effects on rat erythrocytes carbonic anhydrase (CA) were determined. Human erythrocyte carbonic anhydrase isozymes were purified by haemolysate preparation and Sepharose-4B-L tyrosine-sulfanilamide affinity gel chromatography. The HCA-I enzyme, having a specific activity of 7337.5?EU/mg protein, was purified 843-fold with a yield of 60% and the HCA-II enzyme, having a specific activity of 17067?EU/mg protein, was purified 1962-fold with a yield of 22.7%. For in vitro experiments, the enzyme activity was minimal at 2×10-4?M melatonin concentration and increased above this concentration. Ten mg?kg-1 melatonin was administered intraperitoneally and showed a stimulatory effect on the enzyme. Time-dependent in vivo studies were conducted for melatonin in Sprague–Dawley type rats. It was found that CA activity in the rat erythrocytes was decreased by the melatonin after 1 and 3 hours to 2500±500.0 and 1875±239.4 respectively which were statistically significant (p<0.05) differences to the control (2660±235.8). However, CA activity was restored to its normal level after 6?h (2666±235.7) (p>0.05) probably due to metabolism of the melatonin. The findings indicate that melatonin may be pharmacologically useful in some diseases.     

Physical Properties of Mitochondrial Lipids from Lycopersicon hirsutum

Mitochondrial lipids from Lycopersicon hirsutum undergo a broad thermal transition beginning well below 0°C and ending at approximately 25°C. Differential thermal analysis of mitochondrial lipids isolated from ecotypes of L. hirsutum that differ in chilling sensitivity indicates that these lipid preparations have physically similar properties. This was confirmed by electron-spin-resonance experiments, although this technique failed to detect the broad transition detected by differential thermal analysis. No quantitative differences were observed between the percentages of individual lipid classes (based on polar head group) or between the fatty acid compositions of mitochondrial lipids from the two ecotypes investigated. These results suggest that the observed differences between the responses of these ecotypes to prolonged exposure to 5°C may not be related to differences between the physical properties of their mitochondrial lipids.     

Physical and Chemical Properties of an Oncornavirus Associated with a Murine Adrenal Carcinoma Cell Line

A type C oncornavirus has been isolated from a continuous cell line of murine adrenal carcinoma in culture. The particles have a buoyant density of 1.165 g/cm(3), exhibit typical type C morphology by electron microscopy, possess an RNA-dependent DNA polymerase, and have a high molecular weight RNA (6.1 x 10(6)) which can be denatured to a homogeneous lower molecular weight species (3.2 x 10(6)) when extracted from rapidly harvested "immature" virions. The virus is related antigenically to other mammalian oncornaviruses and exhibits a similar, although much more complex, sodium dodecyl sulfate gel electrophoretic profile of virion proteins when compared to the profiles of other type C RNA tumor viruses.     

采用亲和层析结合制备凝胶电泳获得高纯度的重组抗原蛋白用于制备蛋白质芯片

为了得到制备抗原芯片所需的高纯度重组抗原蛋白,需要建立一套适合于多种重组抗原表达和纯化的技术路线．采用了亲和层析结合制备胶电泳的方法,对16种用于构建蛋白质芯片的食管癌相关抗原基因进行了克隆重组并在大肠杆菌中进行了表达．对高表达的重组蛋白首先制备包涵体,然后采用Ni-Sepharose亲和层析得到初步纯化的蛋白质,最后使用SDS-PAGE制备胶电泳作进一步纯化．经过透析复性后,用于制备蛋白质芯片．采用亲和层析纯化重组蛋白,得率为71% ,纯度约为70%;在SDS-PAGE制备胶进一步纯化后,得率为32%,纯度为95%,经过透析和复性后,最终得率为21%,纯度为95%．得到的重组蛋白RPS4在ELISA检测中可以和血清中识别RPS4 的自身抗体起反应,并且,采用精纯抗原制备的蛋白质芯片,在检测抗原与抗体这一对反应中也具有较高的敏感性和特异性,适合大规模血清抗体的检测．研究表明,采用亲和层析结合制备凝胶电泳纯化抗原蛋白,是一条简便快捷,适合需要量不大,但对纯度要求比较高的蛋白质芯片制备的技术路线．     

Effect of Transglutaminase on the Properties of Films Prepared from Chitosan and Gelatin

Applied Biochemistry and Microbiology - The production conditions were optimized, and the characteristics of biodegradable chitosan-gelatin films crosslinked with various microbial...     

Purification and Some Physical Properties of Acid-cellulase from Aspergillus niger

A cellulase preparation which exhibits the highest activity at a lower pH range, 2.3 to 2.5, was purified from a commercial cellulase preparation from a culture filtrate of Asp. niger and referred to as acid-cellulase.

The purification involves ammonium sulfate fractionation, gel filtration and ion-exchange and adsorption chromatographies. The purified enzyme was revealed to be homogenous in ultracentrifugation and disc as well as ampholine electrophoreses and to be an acidic protein of which isoelectric point lied at pH 3.3. The sedimentation coefficient and molecular weight were determined to be 3.27 S and 46,000, respectively. The optical properties were also studied.     

Immunochemical Characteristics and Localization on Cells of Protective Antigen (PAg) Prepared from Leptospira interrogans Serovar lai

Immuno-electron microscopic methods revealed that the protective antigen (PAg) of Leptospira interrogans serovar lai exists on the outer envelope sheathing the leptospiral cell body. PAg lost its protective activity after treatment by hydrolysis with 2 M formic acid at 100 C for 2 hr, or oxidation with periodate at 4 C for 40 hr. The antigenic oligosaccharide fraction was further purified from the hydrolyzed PAg by immunoaffinity column coupled with protective monoclonal antibody, LW2, and by gel filtration of HPLC. The antigenic oligosaccharide fraction contained two unknown sugars and 4-O-methylmannose (molar ratio 3:5:1). These findings suggested that these sugars are components of an antigenic determinant contributing to the protective immunity against serovar lai infection.     

Blastomyces dermatitidis Antibody and Antigen Detection: Comparison of Four Lysate Antigens and Antibodies Prepared from Human Isolates from a Blastomycosis Outbreak

Mycopathologia - Blastomycosis is a systemic fungal disease of humans and other animals produced by the thermally dimorphic fungal organism, Blastomyces dermatitidis. Recent studies have focused on...