Dot-immunobinding assay in the detection of IgG antibodies against farmer's lung antigens

Circulating antibodies against Faenia rectivirgula, Thermoactinomyces candidus, T. vulgaris and Aspergillus fumigatus were studied in the sera of 14 clinically proven farmer's lung patients and 10 normal controls using three immunological methods. These methods were agar gel double diffusion (DD), biotin-avidin-linked immunosorbent assay (BALISA) and dot-immunobinding assay (DIBA). Agar gel diffusion, the least sensitive of the three methods, failed to detect antibodies in some of the patients, while BALISA detected antibodies even in the normal controls. However, the sensitivity of dot-immunobinding assay was in between DD and BALISA while the specificity was comparable to DD to all the antibodies except against A. fumigatus antigens. Dot-immunobinding assay gave faster results than DD and the blots can be stored as record for longer periods of time without fading.     

Aerenchyma and lenticel formation in pine seedlings: A possible avoidance mechanism to anaerobic growth conditions

Seedlings of pond pine ( Pinus serotina Michx.), sand pine [ P. clausa (Engelm.) Sarg.], and loblolly pine ( P. taeda L., wet-site and drought-hardy seed sources) were grown in hydroponic solution culture using a non-circulating, continuously flowing design under anaerobic or aerobic conditions to determine whether flooding tolerance was correlated with enhanced internal root aeration. Transport of atmospheric O₂ from the shoot to the root of anaerobically grown loblolly and pond pine seedlings was demonstrated via rhizosphere oxidation, using both reduced indigo-carmine solution and a polarographic, ensheathing Pt-electrode. Stem and root collar lenticels were the major sites of atmospheric O₂ entry for submerged roots in these seedlings. No O₂ leakage was detected from roots of aerobically grown pine seedlings. Longitudinal and radial pathways for gaseous diffusion via intercellular air spaces in the pericycle and between ray parenchyma cells, respectively, were demonstrated histo-logically in anaerobically grown loblolly and pond pines. Rhizosphere oxidation, and lenticel and aerenchyma development in roots of flood-intolerant sand pine seedlings grown in anaerobic solutions were minimal. Only 15 days of anaerobic growth conditions were necessary to increase internal root porosities of loblolly and pond pine seedlings – although not to the extent found in seedlings treated for 30 or 75 days. Histological results indicated that root tissue in the secondary stage of growth was capable of forming intercellular air spaces, demonstrating a degree of internal plasticity – at least in the more flood-tolerant loblolly and pond pine seedlings.     

Lectin biosynthesis in callus culture established from seeds ofCanavalia virosa

Plant tissue cultures established from isolated embryos or cotyledons were used to investigate the lectin biosynthesis inCanavalia virosa. The lectin (CVL) was detected by double immunodiffusion and hemagglutination tests. CVL was present in all callus cultures. The stem and leaf of the plantlet generated from the embryo also contain CVL, but no CVL was detected in the roots. As compared to mature seeds, callus derived from cotyledon of immature seeds exhibited the largest CVL content. Acknowledgements: The first author would like to express his sincere thanks to Council of Scientific and Industrial Research for the fellowship awarded to him to work on this project.     

Seroimmunological and parasitological surveys of leucocytozoon caulleryi infection in chickens in several asian countries

Epizootiological surveys of Leucocytozoon caulleryi infection in chickens in Japan, Taiwan, Philippines, Singapore, Malaysia and Thailand were undertaken by means of the immunodiffusion test. The rate of infection of L. caulleryi confirmed by the examination of parasites in the peripheral blood of chickens coincided with that of positive antibody response in the immunodiffusion test. Antibodies against L. caulleryi were found in chickens in all the countries surveyed in the present investigation. The prevalence of L. caulleryi infection in chickens was confirmed by the immunodiffusion test. Several chickens in each country showed the presence of serum antigens of L. caulleryi at the times of serum sample collection. These results seemed to indicate that the immunodiffusion test is a method efficient enough to be applicable to the epizootiological surveys and diagnosis of L. caulleryi infection in chickens in the field. As a result, the antibodies or soluble antigens in the sera of chickens infected with L. caulleryi present, respectively, in each country may have the same immunological characters.     

Serum Proteins Promoting [3H]Thymidine Uptake by Trypanosoma (Schizotrypanum) cruzi (Chagas) in vitro

SYNOPSIS. Five proteins capable of stimulating [³H]thymidine uptake by Trypanosoma cruzi in vitro were isolated from fetal calf serum by (NH₄)₂SO₄ precipitation and ion exchange column chromatography. The proteins were partially characterized by immunodiffusion, immunoelectrophoresis, polyacrylamide gel disc electrophoresis, and SDS electrophoresis. As estimated by SDS electrophoresis, using 4 standards, the molecular weight of protein 1 was 100,000, that of protein 2 was 76,000. and that of proteins 3–5 was 68,000 daltons.     

竞争和气候对新疆阿尔泰山西伯利亚五针松树木径向生长的影响

阿尔泰山的北方森林是中亚以及全球的生态系统的重要组成部分, 其生长动态可以影响到全球范围的热辐射、碳平衡等。因此, 探究影响阿尔泰山树木径向生长的主要因素至关重要。该研究以新疆喀纳斯国家级自然保护区的西伯利亚五针松(Pinus sibirica)为研究对象, 建立西伯利亚五针松年表, 通过分析不同时间间隔累年生长量、竞争指数以及气候因子之间的关系, 运用线性混合效应模型、相关分析等方法, 探究竞争和气候对新疆阿尔泰山西伯利亚五针松树木径向生长的影响。结果表明: (1)线性混合效应模型结果显示竞争树胸径和与西伯利亚五针松过去30年的累年生长量之间的拟合效果最好; (2)标准年表与3月的平均气温、平均最高气温、平均最低气温之间有显著正相关关系; (3)累年生长量最高值出现在气温0-5 ℃, 竞争指数低于100的时候。累年生长量最低时, 气温达到-10 ℃, 竞争指数也超过了300。目标树的树木径向生长受到竞争树胸径和及生长季前期气温的影响, 两者共同作用。但相较于气候因子而言, 竞争对西伯利亚五针松的树木径向生长有更大的影响作用。     

Hydroxylation of Phenol to Hydroquinone Catalyzed by A Human Myeloperoxidase-Superoxide Complex: Possible Implications In Benzene-Induced Myelotoxicity

Benzene, a known human rnyelotoxin and leukemogen is metabolized by liver cytochrome P-450 mono-oxygenase to phenol. Further hydroxylation of phenol by cytochrome P-450 monooxygenase results in the formation of mainly hydroquinone, which accumulates in the bone marrow. Bone marrow contains high levels of myeloperoxidase. Here we report that phenol hydroxylation to hydroquinone is also catalyzed by human myeloperoxidase in the presence of a superoxide anion radical generating system, hypoxanthine and xanthine oxidase. No hydroquinone formation was detected in the absence of myeloperoxidase. At low concentrations superoxide disniutase stimulated, but at high concentrations inhibited, the conversion of phenol to hydroquinone. The inhibitory effect at high superoxide dismutase concentrations indicates that the active hydroxylating species of myeloperoxidase is not derived from its interaction with hydrogen peroxide. Furthermore, catalase a hydrogen peroxide scavenger, was found to have no significant effect on hydroxylation of phenol to hydroquinone, supporting the lack of hydrogen peroxide involvement. Mannitol (a hydroxyl radical scavenger) was found to have no inhibitory effect, but histidine (a singlet oxygen scavenger) inhibited hydroquinone formation. Based on these results we postulate that a myeloperoxidase-superoxide complex spontaneously rearranges to generate singlet oxygen and that this singlet oxygen is responsible for phenol hydroxylation to hydroquinone. These results also suggest that myeloperoxidase dependent hydroquinone formation could play a role in the production and accumulation of hydroquinone in bone marrow, the target organ of benzene-induced myelotoxicity.     

Micromanipulation of Gene Expression in the Adult Zebrafish Brain Using Cerebroventricular Microinjection of Morpholino Oligonucleotides

Manipulation of gene expression in tissues is required to perform functional studies. In this paper, we demonstrate the cerebroventricular microinjection (CVMI) technique as a means to modulate gene expression in the adult zebrafish brain. By using CVMI, substances can be administered into the cerebroventricular fluid and be thoroughly distributed along the rostrocaudal axis of the brain. We particularly focus on the use of antisense morpholino oligonucleotides, which are potent tools for knocking down gene expression in vivo. In our method, when applied, morpholino molecules are taken up by the cells lining the ventricular surface. These cells include the radial glial cells, which act as neurogenic progenitors. Therefore, knocking down gene expression in the radial glial cells is of utmost importance to analyze the widespread neurogenesis response in zebrafish, and also would provide insight into how vertebrates could sustain adult neurogenesis response. Such an understanding would also help the efforts for clinical applications in human neurodegenerative disorders and central nervous system regeneration. Thus, we present the cerebroventricular microinjection method as a quick and efficient way to alter gene expression and neurogenesis response in the adult zebrafish forebrain. We also provide troubleshooting tips and other useful information on how to carry out the CVMI procedure.     

Implications of a zinc uptake and transport model

While physicists regularly use mathematical equations to describe natural phenomena, mathematical modeling of biological systems is still not well established and is hampered by communication barriers between experimental and theoretical biologists. In a recent study we developed a mathematical model of zinc uptake and radial transport in Arabidopsis thaliana roots. By refraining from writing many equations in the main text and confining the derivation of formulas to a supplemental file, we attempted to reach both experimentalists and theoreticians likewise. Here, we give a short summary of our results on the accumulation pattern of zinc and the importance of transporter regulation, water flow and geometry. For a better understanding of the dynamics of adaptation to changes in external conditions, we plead for more detailed and frequent measurements. As a new aspect, we analyzed the effect of buffering. Simulations indicate that it dampens oscillations and may therefore play a key role in zinc homeostasis.