Isolation of the inner acrosomal-nuclear membrane complex from rabbit spermatozoa

Rabbit spermatozoa were passed through a Bio-Glas 2500 (BG) column to yield pure cytoplasmic droplets and consequently the droplet-free spermatozoa. Rapid decondensation of sperm nuclei was achieved by 1 mM dithiothreitol (DTT) and 1.5 M NaCl. This treatment caused removal of the plasma and the outer acrosomal membranes. The viscosity of decondensed nuclei was reduced by pancreatic DNase. Further elution of the suspension from BG column yielded a membrane complex. On the basis of electron microscopic observations and the enzyme analysis, these membranes appeared to be the inner acrosomal-nuclear membrane complex (IANC). The IANC was also prepared from isolated sperm head by DTT-NaCl and Dnasetreatment and low-speed centrifugation.     

Thermostable lichenase as a translational reporter

Hybrid genes containing the reporter gene for thermostable lichenase and model genes recA, recA1, cry3a, cry3aM, and ssp1 were constructed. The expression of these genes was studied in prokaryotic and eukaryotic cells. The presence of lichenase in the hybrid proteins was shown to facilitate analysis of the hybrid protein expression in transgenic organisms. Owing to high relative activity and thermostability of lichenase, the activity of this enzyme can be measured by simple, rapid and sensitive qualitative and quantitative methods that do not require costly equipment and reagents. Using the zymograms method, molecular masses of the lichenase-containing hybrid proteins can be precisely estimated. This method is proposed instead of Western blotting using lichenase as a translational reporter. Our results showed that the use of thermostable lichenase as a translational reporter yields the data that are problematic to obtain using traditional methods of gene expression analysis, which is of importance for fundamental and applied research.Translated from Genetika, Vol. 41, No. 1, 2005, pp. 30–39.Original Russian Text Copyright © 2005 by Komakhin, Abdeeva, Salehi Dzhuzani, Goldenkova, Zhuchenko.     

Characterization of a Phytoplasma Associated with Pear Decline in Iran

Symptoms of pear decline (PD) were observed in several pear growing regions of Iran. Pear trees with typical symptoms of PD from Estahban (Fars Province) were examined for phytoplasma infection using polymerase chain reaction (PCR) assay. Graft inoculation of healthy pear trees with scions from diseased trees resulted in production of PD symptoms and transmission of phytoplasma as verified by PCR. Target DNA was amplified from symptomatic pear trees with fO1/rO1, an apple proliferation (AP) group-specific primer pair. Physical and putative restriction fragment length polymorphism (RFLP) analyses of fO1/rO1 primed PCR products showed profiles corresponding to AP group, 16SrX-C subgroup ( Candidatus Phytoplasma pyri). Percent similarity values and phylogenetic analysis of fO1/rO1 primed sequences confirmed that, as a member of AP subclade, Estahban PD phytoplasma has a closer relationship to PD and peach yellow leaf roll phytoplasmas than to AP ( Ca . Phytoplasma mali) and European stone fruit yellows ( Ca . Phytoplasma prunorum) phytoplasmas. This is the first report of PD phytoplasma in the eastern Mediterranean.     

Comparative analysis of cost factors in sturgeon fingerling production in Iranian hatcheries (2000–2004)

Production cost analysis in aquaculture is an essential exercise to assist farm managers. Economic assessment of a farm operation also provides the basis to formulate governmental aquaculture and enhancement policies in many regions. The present study employed questionnaires and interviewed managers while also using governmental statistics to gain insight into production cost variables in Iranian sturgeon hatcheries. Within a decade, production of sturgeon fingerlings for release and stock enhancement increased in Iran to more than 21 million fingerlings by 2004. Costs and contributions of various production factors were determined using data obtained from a questionnaire involving all hatcheries between 2000 and 2004. A team of experts completed the questionnaire data sets while conducting interviews at all sturgeon centres and other related departments. From 2000 to 2004 the contribution of A. persicus was 79% of the total number of sturgeon fingerlings produced followed by A. nudiventris with 7.5% and Huso huso with 6.6%. Among the various expenditures between 2000 and 2004, the costs for permanent and part‐time employees contributed the greatest share of total costs, averaging 44%, with a noticeable declining trend from 51% in 2000 to 36% in 2004. Obtaining and incubating fertilized eggs averaged 22% of total costs, increasing during the same time period from 6 to 35%, respectively. On average, the 2000–2004 production cost for a single sturgeon fingerling was estimated at Rials 1667 (US$ 0.20), increasing from Rials 992 (US$ 0.12) to Rials 2623 (US$ 0.29) over these 4 years. Permanent staff at a hatchery was determined as being the principal cost, followed by costs for obtaining fertilized eggs (including broodstock handling). Over the 5‐year study period the results indicated that costs for part‐time labour declined yearly and, conversely, the costs of obtaining broodstocks as well as fertilizing and incubating eggs increased. Considering the background of hatchery production and stock enhancement of sturgeon species and the results of fishing data, it is possible to arrive at a first estimate of the potential contribution of Persian sturgeon farming to the total catch in Iranian waters; it is assumed that these increases were most likely through stock enhancement.     

Effect of environmental pollution on the proteins, allergenic bands, ontogeny and structure of Avicennia marina (Forsk.) Vierh (Avicenniaceae) pollen grains

In Bushehr province of Iran, Avicennia marina trees have grown in Bordekhoon (Mond Protected Area) and Assaluyeh (Marine National Park of Nayband). Contrary to Bordekhoon, Assaluyeh is a petrochemical region with environmental pollution. This study was aimed to studying protein profiles, allergenic bands, ontogeny, structure and elemental composition of tectum of A. marina pollens in Assaluyeh and Bordekhoon. Pollens were collected from two regions and extracted in PBS, and protein profiles of pollens were determined by sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). As an experimental model, 20 female 6–8-week-old Balb/C mice were divided into two groups. The mice of first and second groups were sensitized by Bordekhoon and Assaluyeh pollen extracts, respectively, and mice serum samples were used for immunoblotting. Pollen characteristics were studied using light and scanning electron microscopes. SDS-PAGE showed some differences between pollen protein profiles of two regions. Immunoblotting assay detected that pollens have two allergenic bands and the protein band at 100 KD is the common allergenic protein in two regions. Light microscopy revealed that the development of anther wall was basic type and some abnormalities were observed in microspores and pollens of Assaluyeh. Scanning electron microscopy studies showed that the apertures in considerable numbers of Assaluyeh pollens were closed. The comparison of elemental composition of pollen tectum between two regions showed that pollens of Assaluyeh have accumulated Cu on their tectum. Results obtained indicated that environmental pollution can affect protein profile, allergenic bands, structure and elemental composition of tectum of A. marina pollens.     

Systematic analysis of gut microbiome reveals the role of bacterial folate and homocysteine metabolism in Parkinson’s disease

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