[Psychophysiological reactivity and personality traits of left- and right-handers during mental stress].

The aim of the present study was to establish whether there is dependence of vascular reactivity to mental stress on the levels of neuroticism and anxiety in left- and right-handers. Thirty-two left-handed and 32 righthanded volunteers (16 males and 16 females in each group) between 18 and 30 years of age were studied. During mental stress both groups demonstrated a "standard" type of response, indicating an increase of the sympathetic-adrenal activity. In left-handers, however, the magnitude of the stress vascular reactivity was significantly higher (p<0.05) in comparison with that of right-handers. In left-handed females a moderate negative correlation between vascular reactivity and the levels of neuroticism (r = -0.39) and trait anxiety (r = -0.47, F = 4.04, p = 0.06) was established. In right-handed males a moderate positive correlation between neuroticism and vascular reactivity (r = 0.35) and significant positive correlation between trait anxiety and vascular reactivity (r = 0.60, F = 7.92, p = 0.01) was found. The results obtained show that vascular reactivity to mental stress is not influenced in a specific to left- and right-handers way by the levels of negative emotionality (neuroticism and trait anxiety).

Enhanced ozone strongly reduces carbon sink strength of adult beech (Fagus sylvatica)--resume from the free-air fumigation study at Kranzberg Forest.

Ground-level ozone (O(3)) has gained awareness as an agent of climate change. In this respect, key results are comprehended from a unique 8-year free-air O(3)-fumigation experiment, conducted on adult beech (Fagus sylvatica) at Kranzberg Forest (Germany). A novel canopy O(3) exposure methodology was employed that allowed whole-tree assessment in situ under twice-ambient O(3) levels. Elevated O(3) significantly weakened the C sink strength of the tree-soil system as evidenced by lowered photosynthesis and 44% reduction in whole-stem growth, but increased soil respiration. Associated effects in leaves and roots at the gene, cell and organ level varied from year to year, with drought being a crucial determinant of O(3) responsiveness. Regarding adult individuals of a late-successional tree species, empirical proof is provided first time in relation to recent modelling predictions that enhanced ground-level O(3) can substantially mitigate the C sequestration of forests in view of climate change.

Variation in morphological and biochemical O3 injury attributes of mature Jeffrey pine within canopies and between microsites.

Crown morphology and leaf tissue chemical and biochemical attributes associated with ozone (O3) injury were assessed in the lower, mid- and upper canopy of Jeffrey pine (Pinus jeffreyi Grev. & Balf.) growing in mesic and xeric microsites in Sequoia National Park, California. Microsites were designated mesic or xeric based on topography and bole growth in response to years of above-average precipitation. In mesic microsites, canopy response to O3 was characterized by thinner branches, earlier needle fall, less chlorotic leaf mottling, and lower foliar antioxidant capacity, especially of the aqueous fraction. In xeric microsites, canopy response to O3 was characterized by higher chlorotic leaf mottling, shorter needles, lower needle chlorophyll concentration, and greater foliar antioxidant capacity. Increased leaf chlorotic mottle in xeric microsites was related to drought stress and increased concurrent internal production of highly reactive oxygen species, and not necessarily to stomatal O3 uptake. Within-canopy position also influenced the expression of O3 injury in Jeffrey pine.

Mechanism of rescue of common p53 cancer mutations by second-site suppressor mutations.

The core domain of p53 is extremely susceptible to mutations that lead to loss of function. We analysed the stability and DNA-binding activity of such mutants to understand the mechanism of second-site suppressor mutations. Double-mutant cycles show that N239Y and N268D act as 'global stability' suppressors by increasing the stability of the cancer mutants G245S and V143A-the free energy changes are additive. Conversely, the suppressor H168R is specific for the R249S mutation: despite destabilizing wild type, H168R has virtually no effect on the stability of R249S, but restores its binding affinity for the gadd45 promoter. NMR structural comparisons of R249S/H168R and R249S/T123A/H168R with wild type and R249S show that H168R reverts some of the structural changes induced by R249S. These results have implications for possible drug therapy to restore the function of tumorigenic mutants of p53: the function of mutants such as V143A and G245S is theoretically possible to restore by small molecules that simply bind to and hence stabilize the native structure, whereas R249S requires alteration of its mutant native structure.

Serum levels of female sex hormones in left-handed and right-handed menopausal women.

The study covered 25 left-handed and 27 right-handed clinically healthy females aged between 47 and 55 years during normal menopause. It was carried out in the third year from the beginning of amenorrhoea. The levels of the follicle-stimulating hormone, luteinising hormone, proclactin, estradiol, and progesterone were determined by means of enzyme-immunological methods with IMX (Abbott, USA) and Serozyme I (Biochem Immunosystem Diagnostica, USA) apparatuses. It was established that in left-handed women the serum concentrations both of the follicle-stimulating hormone and luteinising hormone were significantly higher (P <.001), and those of prolactin, estradiol, and progesterone much lower (P <.001), as compared to the right-handed women.

Complement modulatory activity of bisbenzylisoquinoline alkaloids isolated from Isopyrum thalictroides--I. Influence on classical pathway in human serum.

Eleven bisbenzylisoquinoline alkaloids (BBI) were isolated from the plant Isopyrum thalictroides (L.). Treatment of normal human serum (NHS) with BBI resulted in a diminution of the haemolytic activity of the classical pathway (CP). The mode of action of the main alkaloids isopyruthaline (It1), fangchinoline (It2) and isotalictrine (It3) on CP activation was investigated in vitro. The inhibition was time- and temperature-related and for Itl and It3 depended on the concentration of Ca2+ and Mg2+ ions. It was established that the substances reduced C1 haemolytic activity. It2 and It3 enhanced the complement consumption caused by heat aggregated human IgG (HAGG). The BBI prevented the formation of C3 convertase of the classical pathway. The loss of haemolytic activity was partially restored by the addition of C142 reagent (zymosan-treated guinea pig serum) to alkaloids-treated NHS. The addition of the late components C3-9 (EDTA-treated rat sera) recovered to some extent the haemolytic activity of It1-treated NHS, but not of It2- and It3-treated NHS.

Semirational design of active tumor suppressor p53 DNA binding domain with enhanced stability.

We have designed a p53 DNA binding domain that has virtually the same binding affinity for the gadd45 promoter as does wild-type protein but is considerably more stable. The design strategy was based on molecular evolution of the protein domain. Naturally occurring amino acid substitutions were identified by comparing the sequences of p53 homologues from 23 species, introducing them into wild-type human p53, and measuring the changes in stability. The most stable substitutions were combined in a multiple mutant. The advantage of this strategy is that, by substituting with naturally occurring residues, the function is likely to be unimpaired. All point mutants bind the consensus DNA sequence. The changes in stability ranged from +1.27 (less stable Q165K) to -1.49 (more stable N239Y) kcal mol-1, respectively. The changes in free energy of unfolding on mutation are additive. Of interest, the two most stable mutants (N239Y and N268D) have been known to act as suppressors and restored the activity of two of the most common tumorigenic mutants. Of the 20 single mutants, 10 are cancer-associated, though their frequency of occurrence is extremely low: A129D, Q165K, Q167E, and D148E are less stable and M133L, V203A and N239Y are more stable whereas the rest are neutral. The quadruple mutant (M133LV203AN239YN268D), which is stabilized by 2.65 kcal mol-1 and Tm raised by 5.6 degreesC is of potential interest for trials in vivo.

Glaucine analogues as inhibitors of mouse splenocyte activity.

The inhibitory effect of 15 semi-synthetic analogues of glaucine (1) on the lipopolysaccharide (LPS)-induced and the concanavalin A (Con A)-induced proliferation of mouse splenocytes was compared in vitro. Isoboldine (3), bracteoline (4) and dehydroglaucine (9) showed a significantly higher potency to suppress LPS-induced proliferation than 1, while 7-hydroxy-4-methylglaucine (8), 7-formyldehydroglaucine (11), 7-acetyldehydroglaucine (13), 7-benzoyldehydroglaucine (14), oxoglaucine (15) and glaucine-quinol (16) were less inhibitory. Compounds 3, 4, boldine (5), 15 and 16 surpassed significantly the inhibition expressed by 1 on Con A-induced proliferative response. The effect was equal to the inhibition determined for mitomycin C (Mit C) with both mitogens. In contrast to all others analogues, thaliporphine (2) stimulated splenocyte proliferation in both assays. Antibody response against sheep red blood cells (SRBC) was lowered most strongly by cataline (6), 7-methyldehydroglaucine (10) and 16.

Thermostability and irreversible activity loss of exoglucanase/xylanase Cex from Cellulomonas fimi.

The activity of the beta-1,4-glycanase Cex (EC 3.2.1.91) from Cellulomonas fimi is investigated in connection with its industrial application in cellulose hydrolysis and its potential use in cellosaccharide synthesis. Catalytic activity measurements as a function of temperature, complemented with differential scanning calorimetry (DSC) data, are used to characterize the thermostability of the protein and the influence of interdomain interactions. The data suggest that the enzyme is irreversibly deactivated in one of two possible ways: (1) through a low-temperature route characterized by first-order kinetics; or (2) through a high-temperature route characterized by an initial reversible step followed by an irreversible step. Melting temperatures (Tm) of Cex and p-33 (the isolated catalytic domain of Cex) as estimated by DSC are 64.2 and 64.0 degrees C, respectively, suggesting that the binding and catalytic domains of the protein fold independently. Kinetic parameters (Km, kcat, and kcat/Km) of Cex for the hydrolysis of p-nitrophenyl beta-D-cellobioside (pNPC) were determined at temperatures ranging from 15 to 80 degrees C. As demanded by reversible mass-action thermodynamics, the Tm of Cex in the presence of excess ligand as determined from activity-temperature data is ca. 66.55 degrees C, more than 2 degrees C higher than the Tm for Cex under ligand-free conditions. The effect of temperature on the rate constant has been determined using Arrhenius plots. Combined with irreversible deactivation half-life data and DSC data, the results are used to evaluate a model, based on a theory developed by Hei et al. (1993), for predicting the time-dependent activity and active-state stability of the protein under a range of potential operating conditions.

Thermodynamic stability of wild-type and mutant p53 core domain.

Some 50% of human cancers are associated with mutations in the core domain of the tumor suppressor p53. Many mutations are thought just to destabilize the protein. To assess this and the possibility of rescue, we have set up a system to analyze the stability of the core domain and its mutants. The use of differential scanning calorimetry or spectroscopy to measure its melting temperature leads to irreversible denaturation and aggregation and so is useful as only a qualitative guide to stability. There are excellent two-state denaturation curves on the addition of urea that may be analyzed quantitatively. One Zn2+ ion remains tightly bound in the holo-form of p53 throughout the denaturation curve. The stability of wild type is 6.0 kcal (1 kcal = 4.18 kJ)/mol at 25 degrees C and 9.8 kcal/mol at 10 degrees C. The oncogenic mutants R175H, C242S, R248Q, R249S, and R273H are destabilized by 3.0, 2.9, 1.9, 1.9, and 0.4 kcal/mol, respectively. Under certain denaturing conditions, the wild-type domain forms an aggregate that is relatively highly fluorescent at 340 nm on excitation at 280 nm. The destabilized mutants give this fluorescence under milder denaturation conditions.

Functional brain asymmetry, handedness and age characteristics of climacterium in women.

A total of 1985 women aged between 55 and 65 were distributed into two groups (145 left-handers and 1840 right-handers). They were asked to complete a questionnaire on the appearance of menopause, duration of menopausal transition and age of menopause. In left-handed women a significantly earlier appearance of premenopause was established together with a shorter menopause transition and an earlier occurrence of menopause. These results give grounds for a correlation between handedness, functional brain asymmetry, respectively and the genetically determined fading away of ovary steroidogenesis associated with the appearance and progression of the climacterium. In light of the available literature we assume that progressive reduction in ovarian function during climacterium is coupled with possible specific functioning of the hypothalamo-pituitary-gonadal axis, dependent on the type of hemispheric asymmetry.

Functional brain asymmetry, handedness and menarcheal age.

Functional brain asymmetry influences many functions of the organism; the neuroendocrine axis is one that has received insufficient attention. In this study we set us as the goal of studying the link between functional brain asymmetry and menarcheal age in females with left versus right manual dominance. The appearance of the first menarche was used as a natural model of functioning of the hypothalamic-pituitary-gonadal (HPG) axis. 1695 females, aged between 16 and 25 years, were interviewed by questionnaire about manual dominance and menarcheal age. 182 women were selected and divided into 2 groups: all left-handers (n = 91), and a control group of 91 right-handers. We found a significantly lower average age of menarcheal appearance in the left-handers' age of 12.09 +/- 0.16 years compared to the right-handers' age of 13.32 +/- 0.12 years (p < 0.001). The earliest menarcheal age in left-handers was 8 years and the peak of appearance at age 13 (in 30.76% of the cases). In right-handers these values were 10 and 14 years (in 40.60% of the cases), respectively. The data allow us to accept the existence of a link between functional brain asymmetry and menarche, which causes earlier activation of the HPG axis in left-handed females.

[The functional and morphological changes in the liver and kidneys of white rats treated with aluminum]. / Funktsionalni i morfologichni promeni v cherniia drob i bubretsite na beli plukhove, tretirani s aluminii.

A total of 126 white male Wistar rats under subacute conditions were studied. Of them, 76 were perorally treated with AlCl3 in a dose of 3 mg/kg b.w. daily for 40 days. Activities of acid phosphatase (AP), succinate dehydrogenase (SDH), adenosine triphosphatase (ATP-ase) and the contents of glycogen, glucoproteins and RNA were dynamically followed-up in the liver and kidney. Serum activities of ASAT and ALAT were estimated by Borhringer's tests. Morphological changes were histochemically investigated. There was an initial elevation followed by reduction mostly manifested for SDH and ATP-ase and an increase of AP at the end of trial. Histochemical data argued for disorders of protein and carbohydrate metabolism. Morphological alterations more outlined in the liver became more severe until the end of the experiment.

Whole cell biocatalysis in nonconventional media.

In this paper biocatalytic reactions carried out by whole cells in nonconventional media are reviewed. Similar relationships are observed between solvent hydrophobicity and catalytic activity in reactions carried out by isolated enzymes and whole cells. In addition to the effect of organic solvent on biocatalyst stability, microbial cells are susceptible to damaging effects caused by the organic phase. In general, more hydrophobic solvents manifest lower toxicity towards the cells. Whole cell biocatalysts require more water than isolated enzymes and two-phase systems have been most widely used to study whole cell biocatalysis. Immobilization makes cell biocatalysts more resistant to organic solvents and helps achieve homogeneous biocatalyst dispersion. Cell entrapment methods have been widely used with organic solvent systems and mixtures of natural and/or synthetic polymers allow adjustment of the hydrophobicity-hydrophilicity balance of the support matrix. Some examples of stereoselective catalysis using microbial cells in organic solvent media are presented.

Reductive biotransformation by wild type and mutant strains of Saccharomyces cerevisiae in aqueous-organic solvent biphasic systems.

Whole cells of Saccharomyces cerevisiae analyzed the conversion of benzaldehyde to benzyl alcohol in aqueous-organic biphasic media. Reaction rate increased dramatically as moisture content of the solvent was increased in the range 0% to 2%. The highest biotransformation rates were observed when hexane was used as organic solvent. Benzaldehyde was also converted to benzyl alcohol by a cell-free crude extract in biphasic systems containing hexane, although the rate of product formation was much lower. Mutant strains of S. cerevisiae lacking some or all of the ADH isoenzymes, ADH I, II, and III, manifested similar rates for bioconversion of benzaldehyde to benzyl alcohol in both aqueous and two-phase systems. In general, conversion rates observed in aqueous media were 2 to 3 times higher than those observed in hexane containing 2% moisture.

Production of L-phenylacetyl carbinol by biotransformation: product and by-product formation and activities of the key enzymes in wild-type and ADH isoenzyme mutants of Saccharomyces cerevisiae.

The capacities of yeast wild-type and mutants strains known to lack specific ADH isoenzymes to produce L-phenylacetyl carbinol (PAC) and benzyl alcohol in biotransformation trials were also investigated. Pyruvate decarboxylase activity, responsible for PAC formation and ADH activity, which can participate in reduction of benzaldehyde to benzyl alcohol, was also determined in each strain. In addition, the capacity of each strain to produce ethanol was investigated. Mutant strains lacking all of the isoenzymes, ADH-I, ADH-II, and ADH-III, still exhibited some ADH activity and were capable of production of benzyl alcohol and ethanol.

The effect of certain heavy metals (Mn and Pb) on parameters of erythrocyte energy metabolism.

Two groups of workers without clinical manifestations of intoxications were examined in the study. Workers in Group One were occupationally exposed to manganese (Mn) and in Group Two - to lead (Pb). Erythrocyte concentrations of adenyl nucleotides (AMP, ADP and ATP) as well as that of inorganic phosphorus (Pi) were determined. The authors also calculated both ratios ADP/Pi and ATP/ADP, the energy charge and phosphorylation potential. The metabolic processes in the erythrocytes of workers exposed to heavy metals but without clinical manifestations were found to intensify and become more pronounced in workers occupationally exposed to lead. Erythrocytes from workers occupationally exposed to lead featured higher ratios of ATP/ADP.