Identification of differential gene expression for <Emphasis Type="Italic">Kr1</Emphasis> gene in bread wheat using annealing control primer system

Intergeneric hybridization is an important strategy to introgress alien genes into common wheat for its improvement. But presence of cross ability barrier mechanism regulated by Kr1 gene played a major destructive role for hybridization than other reported genes. In order to know the underlying molecular mechanism and to dissect out this barrier, a new annealing system, ACP (anneling control primer) system was used in chromosome 5B (containing Kr1 gene) specific Recombinant Inbred Line (RIL) population. Two differentially expressed fragments for Kr1 gene was identified, cloned and sequenced. Further the expression was confirmed by northern blotting analysis. Sequence analysis of the resulted clones revealed classes of putative genes, including stress responsive and signal transduction.     

Recent achievements in in vitro culture and preservation of ovarian follicles in mammals

The mammalian ovary contains a large number of follicles that are in various developmental stages. The largest portion of them are primordial follicles. However, throughout the female reproductive lifespan only a small proportion of these follicles will produce oocytes competent to undergo successful maturation and ovulation. The rest of the ovarian oocytes (>99.9%) undergo atresia. It would be of great practical benefit to rescue some of these follicles by growing them in culture in order to provide an extra source of gametes. There is considerable interest in developing technologies that aim to produce fully-grown, developmentally competent oocytes from a pool of early developmental stages of follicles. Two methods have been used: 1/ long-term in vitro culture of either follicles or oocytes, and 2/ transplantation of ovarian tissue grafts. The development of efficient technologies may provide an additional source of oocytes for livestock production and reproduction in humans and rare or endangered species. The aim of this paper is to present a comprehensive review of recent achievements in the utilization of small ovarian follicles (primordial, preantral and early antral) by long-term in vitro culture and/or transplantation of ovarian tissue grafts (fresh and cryopreserved) in mammals including humans.     

A new type of cell in the pulmonary epithelium of the newt Triturus alpestris Laur

Summary Single cells of a new type appear scattered among pneumocytes in the pulmonary epithelium. The surfaces of these cells communicate with the air space and display numerous finger-like microvilli. In comparison to pneumocytes, these cells have a more lucid cytoplasm and their apical parts contain large amounts of electron-lucent vesicles and electron-dense granules, which are probably released into the lumen of the lung. These secretory cells exhibit a yellow formaldehyde-induced fluorescence, which suggests that they belong to the class of APUD cells.     

Kinetic model of oxidation catalyzed by xanthine oxidase-the final enzyme in degradation of purine nucleosides and nucleotides

A new kinetic model is presented for analysis of experimental data of oxidation process catalyzed by milk xanthine oxidase. The kinetics for two substrates, xanthine and its analog 2-chloroadenine, in a broad pH range (5.8-9.0) are best described by an equation which is a rational function of degree 2:3 and 2:2, respectively.     

Design of selective substrates of proteinase 3 using combinatorial chemistry methods

In this study, chemical synthesis of the selective chromogenic/fluorogenic substrates for proteinase 3 is described. The substrates’ sequence was obtained using combinatorial chemistry methods. Deconvolution of the tripeptide library against proteinase 3 with general formula ABZ-X₃-X₂-X₁-ANB-NH₂ yielded the active sequence. Selected peptide was further modified on its C terminus to investigate the impact of chromophore moiety modification on enzyme-substrate interaction. To determine specificity, activity of selected substrates was characterized against proteinase 3 and neutrophil elastase. Finally, the peptide ABZ-Tyr-Tyr-Abu-ANB-NH₂ displayed the highest value of specificity constant (k_cat/K_M = 189 × 10³ M⁻¹ s⁻¹) for proteinase 3. To the best of our knowledge, this is the first short peptide that undergoes selective proteolysis by proteinase 3 and displays no significant hydrolysis in the presence of human neutrophil elastase and cathepsin G.     

Recycle use of immobilized glucoamylase by tangential flow filtration unit

Various properties of glucoamylase immobilized onto corn stover supporting material and separation of immobilized enzyme by tangential flow filtration unit were studied. Optimum pH and temperature of immobilized enzyme were 3.5 and 60 degrees C, respectively. Enzyme stability was studied in a packed-bed column. The starch conversion rate was attained at 81% for 15 days; after that, the hydrolysis rate gradually decreased. Size of supporting material proved to be an important factor, with higher activity and good loading yield resulting from smaller supporting material. Glucoamylase immobilized onto supporting material less than 44 mum was used for hydrolysis of 10% soluble starch at pH 3.5 and 40 degrees C for 3 h. Then immobilized glucoamylase was separated from the product by means of a tangential flow filtration unit using a 0.2-mum pore size Nylon 66 membrane filter. This operation was continued until 180 ml filtrate was obtained from a 260-mL starting volume. Then, the next batch was started by adding 180 mL starch substrate into the reactor. The batchwise experiments were repeated 20 times. The average filtration rate of each batch was determined and found to sharply decline during the first four batches. Thereafter, it gradually decreased from batch to batch. The cause of decreasing filtration rate appeared to be due to retrogradation of starch. The percentage of starch hydrolysis within 20 batches was in the range 89-96%. The filtration rate becomes higher if the hydrolyzation time is extended to 14 h. Resistance to filtration was also investigated. Almost all of the total resistance is related to insoluble materials, with the significant part of this from the resistance due to insoluble materials deposited on a surface of membrane and boundary layer resistance. Using a microscopic method, no microorganisms were found in the filtrate.     

Methylmercury and Total Mercury in Eels, <Emphasis Type="Italic">Anguilla anguilla</Emphasis>, from Lakes in Northeastern Poland: Health Risk Assessment

In the aquatic environment, mercury is readily methylated into its most toxic form of methylmercury. In this form, it enters the aquatic food chain and its concentrations increase in subsequent links, which decreases the quality of fish meat and poses risks to consumer health. Concentrations of methylmercury (MeHg) and total mercury (THg) were determined in the muscle tissues of 64 eel specimens measuring from 59 to 95 cm in length as functions of specimen size and weight. Risks posed to consumers by eel from different length classes were also assessed. The mean concentration of THg in all of the eel examined was 0.179 mg kg^?1, but the range was from 0.028 to 0.487 mg kg^?1. The mean concentration of MeHg was 0.147 mg kg^?1, and the range was also wide from 0.023 to 0.454 mg kg^?1. Accumulated MeHg and THg increased with eel body length. The percentage share of MeHg in THg also changed with specimen length, and there was a positive correlation between the concentrations of MeHg and THg. Risk assessment was performed based on the doses of THg and MeHg ingested with fish for several specimen length classes. Consuming the meat of eel measuring 80 cm in length increased the estimated weekly intake (EWI) of THg and MeHg twofold in comparison to that from specimens 60 cm in length and fourfold in specimens exceeding 90 cm in length. The percentage shares of the EWI in the tolerable weekly intake and the target hazard quotient coefficient also increased proportionally. Generally, concentrations of MeHg and THg in eel are below current limits and pose no risk to consumer health as long as the consumption of larger specimens is avoided.     

Molecular simulations of intermediate and long alkanes adsorbed on graphite: Tuning of non-bond interactions

The interplay between the torsional potential energy and the scaling of the 1-4 van der Waals and Coulomb interactions determines the stiffness of flexible molecules. In this paper we demonstrate for the first time that the precise value of the nonbond scaling factor (SF)—often a value assumed without justification—has a significant effect on the critical properties and mechanisms of systems undergoing a phase transition, and that, for accurate simulations, this scaling factor is highly dependent on the system under consideration. In particular, by analyzing the melting of n-alkanes (hexane C6, dodecane C12, tetracosane C24) on graphite, we show that the SF is not constant over varying alkane chain lengths when the structural correlated transformations are concerned. Instead, monotonic decrease of SF with the molecular length drives a cross-over between two distinct mechanisms for melting in such systems. In a broad sense we show that the choice for SF in any simulation containing adsorbed or correlated long molecules needs to be carefully considered.