Regulation of DNA replication by homopurine/homopyrimidine sequences

The simple repeating homopurine/homopyrimidine sequences dispersed throughout many eukaryotic genomes are known to form triple helical structures comprising three-stranded and single-stranded DNA. Several lines of evidence suggest that these structures influence DNA replication in cells. Homopurine/homopyrimidine sequences cloned into simian virus 40 (SV40) or SV40 origin-containing plasmids caused a reduced rate of DNA synthesis due to the pausing of replication forks. More prominent arrests were observed in in vitro experiments using single-stranded and double-stranded DNA with triplex-forming sequences. Nucleotides unable to form triplexes when present in the template DNA or when incorporated into the nascent strand prevented termination. Similarly, mutations destroying the triplex potential did not cause arrest while compensatory mutations restoring triplex potential restored it. These and other observations from a number of laboratories indicating that homopurine/homopyrimidine sequences act as arrest signals in vitro and as pause sites in vivo during replication fork movement suggest that these naturally occurring sequences play a regulatory role in DNA replication and gene amplification.     

Differential expression of Alpha,Mu, and Pi classes of glutathione S-transferases in chemosensory mucosae of rats during development

The expression of three classes of glutathione S-transferases (GSTs), Alpha, Mu, and Pi was investigated in the nasal mucosae of rats during development using immunohistochemical methods. GST Alpha and Mu were first detected in the supranuclear region of sustentacular cells on embryonic days 16. The Bowman's glands expressed differential patterns of immunoreactivity during development, beginning at postnatal day (P) 2 and P6 for Alpha and Mu classes, respectively and being greatest at P11 for both. The acinar cells of vomeronasal glands in the vomeronasal organ expressed Alpha and Mu classes of GSTs from P11 onwards. In the septal organ of Masera, the supranuclear region of sustentacular cells expressed GSTs from P11 with little or no variation during development. In the respiratory mucosa, Alpha and Mu classes of GSTs were detected at the brush borders of ciliated cells and in the acinar cells of posterior septal glands, but not in anterior septal or respiratory glands located on the turbinates. Compared to olfactory mucosa, the changes in immunoreactivity for GSTs were less pronounced in the respiratory mucosa during development. Specific GST Pi immunoreactivity was not detected in the nasal mucosae at any stage of development studied. The occurrence of GSTs in the nasal mucosa, including olfactory, vomeronasal, septal, and respiratory epithelia, suggests that the GSTs are actively involved in the biotransformation of xenobiotics including odorants and pheromones, and may also participate in perireceptor processes such as odorant clearance. In addition, we have developed a working model describing the cellular localization of certain phase I (e.g., cytochrome P-450s) and phase II (e.g., GSTs, -glutamyl transpeptidase) biotransformation enzymes in the olfactory mucosa and their proposed roles in xenobiotic metabolism.     

Severe hypoxia decreases oxygen uptake relative to intensity during submaximal graded exercise

The effect of severe acute hypoxia (fractional concentration of inspired oxygen equalled 0.104) was studied in nine male subjects performing an incremental exercise test. For power outputs over 125 W, all the subjects in a state of hypoxia showed a decrease in oxygen consumption ( O₂) relative to exercise intensity compared with normoxia (P < 0.05). This would suggest an increased anaerobic metabolism as an energy source during hypoxic exercise. During submaximal exercise, for a given O₂, higher blood lactate concentrations were found in hypoxia than in normoxia (P < 0.05). In consequence, the onset of blood lactate accumulation (OBLA) was shifted to a lower O₂ ( O₂ 1.77 l·min^–1 in hypoxia vs 3.10 l·min^–1 in normoxia). Lactate concentration increases relative to minute ventilation ( _E) responses were significantly higher during hypoxia than in normoxia (P < 0.05). At OBLA, _E during hypoxia was 25% lower than in the normoxic test. This study would suggest that in hypoxia subjects are able to use an increased anaerobic metabolism to maintain exercise performance.     

Comparative study of the genetic effect of various tritium compounds in male mice

Comparative study of the genetic effect of some tritiated biogenic++ compounds and tritiated water was carried out. Reciprocal translocations induced in mice stem spermatogonia by 3H-glucose, 3H-glycine and 3H-lysine occur with similar frequency, whereas the difference in values of the absorbed doses of beta-irradiation in some cases was about two-fold. The highest genetic effect per unit of activity of 3H-nucleosides (3H-TdR and 3H-CdR) was revealed at lowest activities. The frequency of reciprocal translocations induced by 3H-TdR was twice as that induced by 3H-CdR.     

Skeletal troponin I as a marker of exercise-induced muscle damage

Sorichter, Stephan, Johannes Mair, Arnold Koller, WalterGebert, Daniel Rama, Charles Calzolari, Erika Artner-Dworzak, and BerndPuschendorf. Skeletal troponin I as a marker of exercise-inducedmuscle damage. J. Appl. Physiol.83(4): 1076-1082, 1997.The utility of skeletal troponin I (sTnI)as a plasma marker of skeletal muscle damage after exercise wascompared against creatine kinase (CK), myoglobin (Mb), and myosin heavychain (MHC) fragments. These markers were serially measured in normalphysical education teacher trainees after four different exerciseregimens: 20 min of level or downhill (16% decline) running(intensity: 70% maximal O₂uptake), high-force eccentric contractions (70 repetitions), orhigh-force isokinetic concentric contractions of the quadriceps group(40 repetitions). Eccentrically biased exercise (downhill running andeccentric contractions) promoted greater increases in all parameters.The highest plasma concentration were found after downhill running{median peaks: 309 U/l CK concentration ([CK])}, 466 µg/l Mb concentration([Mb]), 1,021 µU/l MHC concentration ([MHC]),and 27.3 µg/l sTnI concentration ([sTnI]). Level running produced a moderate response (median peaks: 178 U/l [CK],98 µg/l [Mb], 501 µU/l [MHC], and 6.6 µg/l [sTnI]), whereas the concentric contraction protocoldid not elicit significant changes in any of the markers assayed. sTnIincreased and peaked in parallel to CK and stayed elevated (>2.2µg/l) for at least 1-2 days after exercise. In contrast to MHC,sTnI is an initial, specific marker of exercise-induced muscle injury,which may be partly explained by their different intracellularcompartmentation with essentially no (MHC <0.1%) or a small solublepool (sTnI: median 3.4%).

     

Enhanced plant regeneration in grain and sweet sorghum by asparagine,proline and cefotaxime

Summary Cefotaxime ( 50 and 100 mg/1 ), a cephalosporin antibiotic and the amino acids asparagine and proline (200 mg/l) enhanced the production of embryogenic callus, increased the frequency of plant regeneration, and delayed the loss of regeneration potential in immature embryo-derived callus cultures ofSorghum bicolor (L.) Moench. Although these compounds did not promote callus induction or growth of callus, they influenced plant regeneration considerably in 10 low responding genotypes of grain and high anthocyanin containing sweet sorghums.     

Biomass protein formation by filamentous fungi on woody substrates

Summary Growth and biomass protein formation by filamentous fungi grown on pretreated tropical woods of Mesta (Hibiscus cannabinus Linn.) and Subabul [Leucaena leucocephala (Lam.) de Witt] as well as their isolated hemicellulose and cellulose fractions have been studied. Penicillium janthinellum and Penicillium funiculosum produced a biomass having 20 to 30% crude protein when grown on either hemicellulose, while growth on pretreated (autoclaved in 1% NaOH) wood or isolated cellulose fractions was comparatively poor and crude protein content only 5 to 8% in the biomass.NCL Communication no.3550     

Phospholipid vesicle aggregation induced by human myelin basic protein

Human myelin basic protein isolated from the brains of individuals who died with multiple sclerosis was more potent in inducing the aggregation of egg phosphatidylcholine vesicles than was the basic protein isolated from the brains of normal individuals. The portion of myelin basic protein which bound to egg phosphatidylcholine vesicles was separated from the free protein by sucrose density gradient centrifugation. Similar amounts of basic protein from normal or from multiple sclerosis brains are bound to the lipid and no consistent differences in the N^G, N^G dimethyl-arginine content of the protein fractions have been found.     

A general multistate model for the analysis of hydrogen-exchange kinetics

In proton nmr, the chemical exchange rates of slowly exchanging labile hydrogens (with lifetimes in the range ～ 10 msec – ～ 1 sec) of peptides, proteins, and nucleic acids can be measured in H₂O by a combination of two separate experiments: (1) the transfer of solvent saturation and (2) saturation-recovery experiments. When these molecules exist in a dynamic equilibrium among different conformations, the experiments cannot be analyzed in a straightforward manner to derive the intrinsic exchange rates. In the present study we have derived analytical expressions for the above two experiments on a biomolecule under certain limiting conditions: (1) the extreme low-motility limit, where each of the conformational transitions is much slower than the corresponding hydrogen exchange rate with the solvent; (2) the high-motility limit (EX₂ mechanism), which is the opposite extreme of the previous limit; and (3) the low-motility limit (EX₁ mechanism), which is a mixture of limits (1) and (2), i.e., for some of the conformations, the exchange rate with the solvent is much faster than their conformational transition rates, while for the remaining conformations the reverse situation is realized. The results may be considered as a generalization to an arbitrary number of states of the two-state model treated by Hvidt. Equations have also been derived that are applicable to the iostope exchange method of measuring very slow exchange rates (with life-times of the order of minutes and longer) in biomolecules. The saturation recovery experiments performed in H₂O on the active pentapeptide fragment of thymopoietin serve to illustrate the high-motility limit. The theoretical formulation presented in this study can be easily adapted to other double-resonance techniques and also to situations where the kinetics of an arbitrary system existing in a multistate equilibrium are of interest.