In vitro–in vivo study on the effects of plant compounds on rumen fermentation,microbial abundances and methane emissions in goats

Two in vitro and one in vivo experiments were conducted to investigate the effects of a selection of plant compounds on rumen fermentation, microbial concentration and methane emissions in goats. Treatments were: control (no additive), carvacrol (CAR), cinnamaldehyde (CIN), eugenol (EUG), propyl propane thiosulfinate (PTS), propyl propane thiosulfonate (PTSO), diallyl disulfide (DDS), a mixture (40 : 60) of PTS and PTSO (PTS+PTSO), and bromochloromethane (BCM) as positive control with proven antimethanogenic effectiveness. Four doses (40, 80, 160 and 320 µl/l) of the different compounds were incubated in vitro for 24 h in diluted rumen fluid from goats using two diets differing in starch and protein source within the concentrate (Experiment 1).The total gas production was linearly decreased (P<0.012) by all compounds, with the exception of EUG and PTS+PTSO (P⩾0.366). Total volatile fatty-acid (VFA) concentration decreased (P⩽0.018) only with PTS, PTSO and CAR, whereas the acetate:propionate ratio decreased (P⩽0.002) with PTS, PTSO and BCM, and a tendency (P=0.064) was observed for DDS. On the basis of results from Experiment 1, two doses of PTS, CAR, CIN, BCM (160 and 320 µl/l), PTSO (40 and 160 µl/l) and DDS (80 and 320 µl/l) were further tested in vitro for 72 h (Experiment 2). The gas production kinetics were affected (P⩽0.045) by all compounds, and digested NDF (DNDF) after 72 h of incubation was only linearly decreased (P⩽0.004) by CAR and PTS. The addition of all compounds linearly decreased (P⩽0.009) methane production, although the greatest reductions were observed for PTS (up to 96%), DDS (62%) and BCM (95%). No diet–dose interaction was observed. To further test the results obtained in vitro, two groups of 16 adult non-pregnant goats were used to study in vivo the effect of adding PTS (50, 100 and 200 mg/l rumen content per day) and BCM (50, 100 and 160 mg/l rumen content per day) during the 9 days on methane emissions (Experiment 3). The addition of PTS and BCM resulted in linear reductions (33% and 64%, respectively, P⩽0.002) of methane production per unit of dry matter intake, which were lower than the maximum inhibition observed in vitro (87% and 96%, respectively). We conclude that applying the same doses in vivo as in vitro resulted in a proportional lower extent of methane decrease, and that PTS at 200 mg/l rumen content per day has the potential to reduce methane emissions in goats. Whether the reduction in methane emission observed in vivo persists over longer periods of treatments and improves feed conversion efficiency requires further research.     

Divergent effects of compounds on the hydrolysis and transpeptidation reactions of γ-glutamyl transpeptidase

A novel class of inhibitors of the enzyme γ-glutamyl transpeptidase (GGT) were evaluated. The analog OU749 was shown previously to be an uncompetitive inhibitor of the GGT transpeptidation reaction. The data in this study show that it is an equally potent uncompetitive inhibitor of the hydrolysis reaction, the primary reaction catalyzed by GGT in vivo. A series of structural analogs of OU749 were evaluated. For many of the analogs, the potency of the inhibition differed between the hydrolysis and transpeptidation reactions, providing insight into the malleability of the active site of the enzyme. Analogs with electron withdrawing groups on the benzosulfonamide ring, accelerated the hydrolysis reaction, but inhibited the transpeptidation reaction by competing with a dipeptide acceptor. Several of the OU749 analogs inhibited the transpeptidation reaction by slow onset kinetics, similar to acivicin. Further development of inhibitors of the GGT hydrolysis reaction is necessary to provide new therapeutic compounds.     

Induced expression of the IER5 gene by γ-ray irradiation and its involvement in cell cycle checkpoint control and survival

The immediate-early response gene 5 (IER5) was previously shown, using microarray analysis, to be upregulated by ionizing radiation. Here we further characterized the dose- and time-dependency of radiation-induced expression of IER5 at doses from 0.5 to 15 Gy by quantitative real-time PCR analyses in HeLa cells and human lymphoblastoid AHH-1 cells. A radiation-induced increase in the IER5 mRNA level was evident 2 h after irradiation with 2 Gy in both cell lines. In AHH-1 cells the expression reached a peak at 4 h and then quickly returned to the control level, while in HeLa cells the expression only remained increased for a short period of time at around 2 h after irradiation before returning to the control. After high-dose irradiation (10 Gy), the induction of the IER5 expression was lower and delayed in AHH-1 cells as compared with 2-Gy irradiated cells. In HeLa cells, at this dose, two peaks of increased expression were observed 2 h and 12–24 h post-irradiation, respectively. RNA interference technology was employed to silence the IER5 gene in HeLa cells. siRNA-mediated suppression of IER5 resulted in an increased proliferation of HeLa cells. Cell growth and survival analyses demonstrated that suppression of IER5 significantly increased the radioresistance of HeLa cells to radiation doses of up to 6 Gy, but barely affected the sensitivity of cells at 8 Gy. Moreover, suppression of IER5 potentiated radiation-induced arrest at the G2-M transition and led to an increase in the fraction of S phase cells. Taken together, we propose that the early radiation-induced expression of IER5 affects the radiosensitivity via disturbing radiation-induced cell cycle checkpoints.     

Inputs of seabird guano alter microbial growth,community composition and the phytoplankton–bacterial interactions in a coastal system

Seabird guano enters coastal waters providing bioavailable substrates for microbial plankton, but their role in marine ecosystem functioning remains poorly understood. Two concentrations of the water soluble fraction (WSF) of gull guano were added to different natural microbial communities collected in surface waters from the Ría de Vigo (NW Spain) in spring, summer, and winter. Samples were incubated with or without antibiotics (to block bacterial activity) to test whether gull guano stimulated phytoplankton and bacterial growth, caused changes in taxonomic composition, and altered phytoplankton–bacteria interactions. Alteromonadales, Sphingobacteriales, Verrucomicrobia and diatoms were generally stimulated by guano. Chlorophyll a (Chl a) concentration and bacterial abundance significantly increased after additions independently of the initial ambient nutrient concentrations. Our study demonstrates, for the first time, that the addition of guano altered the phytoplankton–bacteria interaction index from neutral (i.e. phytoplankton growth was not affected by bacterial activity) to positive (i.e. phytoplankton growth was stimulated by bacterial activity) in the low-nutrient environment occurring in spring. In contrast, when environmental nutrient concentrations were high, the interaction index changed from positive to neutral after guano additions, suggesting the presence of some secondary metabolite in the guano that is needed for phytoplankton growth, which would otherwise be supplied by bacteria.     

Different effects of α- and γ-glycine on the aberrant activity of pyramidal neurons of hippocampal slices

Experiments in vitro on hippocampal slices of mouse have shown that solutions prepared from polymorphic modifications α- and γ-glycine have different effect on the aberrant activity of neurons. In the presence of α-glycine the excitability of these neurons decreased more slowly, prolonging its modulating effect on NMDA type glutamate receptors. This effect agrees with higher biological activity of α-polymorphic modifications (as compared with that of the α-form) that previously observed with respect to behavior of mice from the line with genetic diathesis to catalepsy, which were used as a biological model for investigation of some pathological behavior forms.     

Evaluation of the effects and interactions of mixing and oxygen transfer on the production of Fab’ antibody fragments in Escherichia coli fermentation with gas blending

Fermentations carried out at 450-L and 20-L scale to produce Fab’ antibody fragments indicated a serious problem to control levels of dissolved oxygen in the broth due to the large oxygen demand at high cell densities. Dissolved oxygen tension (DOT) dropped to zero during the induction phase and it was hypothesised that this could limit product formation due to inadequate oxygen supply. A gas blending system at 20-L scale was employed to address this problem and a factorial 2² experimental design was executed to evaluate independently the effects and interaction of two main engineering factors: agitation rate and DOT level (both related to mixing and oxygen transfer in the broth) on Fab’ yields. By comparison to the non-gas blending system, results in the gas blending system at same scale showed an increase in the production of Fab’ by 77% independent of the DOT level when using an agitation rate of 500 rpm level and by 50% at an agitation rate of 1,000 rpm with 30% DOT. Product localisation in the cell periplasm of >90% was obtained in all fermentations. Results obtained encourage further studies at 450-L scale initially, to evaluate the potential of gas blending for the industrial production of Fab’ antibody fragments.     

Elevation of antioxidant potency in the brain of mice by low-dose γ-ray irradiation and its effect on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced brain damage

The elevation of endogenous thiol-related antioxidants and free radical scavenging enzymes in the brain of C57BL/6 female mice after low-dose γ-ray irradiation and its inhibitory effect on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced brain damage were investigated. The brain level of the reduced form of glutathione (GSH) increased soon after irradiation with 50 cGy of γ-rays, reached a maximum at 3 h post-treatment, and remained elevated until 12 h. Thioredoxin (TRX) was also transiently increased after irradiation. The activities of free radical scavenging enzymes, including Cu/Zn-superoxide dismutase, catalase and glutathione peroxidase, were significantly induced after irradiation as well. Cerebral malondialdehyde was remarkably elevated by MPTP treatment, and this elevation was suppressed by pre- irradiation (50 cGy). The contents of GSH and TRX were significantly decreased by MPTP treatment in comparison with those of the control group. These reductions both seemed to be attenuated by pre-irradiation with γ-rays. These results suggest that low-dose γ-ray irradiation induces endogenous antioxidative potency in the brain of mice and might be effective for the prevention and/or therapy of various reactive oxygen species-related neurodegenerative disorders, such as Parkinson’s disease and Alzheimer’s disease.     

Pentoxifylline enhances the radioprotective properties of γ-tocotrienol: differential effects on the hematopoietic, gastrointestinal and vascular systems

The vitamin E analog γ-tocotrienol (GT3) is a potent radioprotector and mitigator. This study was performed to (a) determine whether the efficacy of GT3 can be enhanced by the addition of the phosphodiesterase inhibitor pentoxifylline (PTX) and (b) to obtain information about the mechanism of action. Mice were injected subcutaneously with vehicle, GT3 [400 mg/kg 24 h before total-body irradiation (TBI)], PTX (200 mg/kg 30 min before TBI), or GT3+PTX before being exposed to 8.5-13 Gy TBI. Overall lethality, survival time and intestinal, hematopoietic and vascular injury were assessed. Cytokine levels in the bone marrow microenvironment were measured, and the requirement for endothelial nitric oxide synthase (eNOS) was studied in eNOS-deficient mice. GT3+PTX significantly improved survival compared to GT3 alone and provided full protection against lethality even after exposure to 12.5 Gy. GT3+PTX improved bone marrow CFUs, spleen colony counts and platelet recovery compared to GT3 alone. GT3 and GT3+PTX increased bone marrow plasma G-CSF levels as well as the availability of IL-1α, IL-6 and IL-9 in the early postirradiation phase. GT3 and GT3+PTX were equally effective in ameliorating intestinal injury and vascular peroxynitrite production. Survival studies in eNOS-deficient mice and appropriate controls revealed that eNOS was not required for protection against lethality after TBI. Combined treatment with GT3 and PTX increased postirradiation survival over that with GT3 alone by a mechanism that may depend on induction of hematopoietic stimuli. GT3+PTX did not reduce GI toxicity or vascular oxidative stress compared to GT3 alone. The radioprotective effect of either drug alone or both drugs in combination does not require the presence of eNOS.     

Using energetic budgets to assess the effects of environmental stress on corals: are we measuring the right things?

Historically, the response of marine invertebrates to their environment, and environmentally induced stress, has included some measurement of their physiology or metabolism. Eventually, this approach developed into comparative energetics and the construction of energetic budgets. More recently, coral reefs, and scleractinian corals in particular, have suffered significant declines due to climate change-related environmental stress. In addition to a number of physiological, biophysical and molecular measurements to assess “coral health,” there has been increased use of energetic approaches that have included the measurement of specific biochemical constituents (i.e., lipid concentrations) as a proxy for energy available to assess the potential outcomes of environmental stress on corals. In reading these studies, there appears to be some confusion between energy budgets and carbon budgets. Additionally, many assumptions regarding proximate biochemical composition, metabolic fuel preferences and metabolic quotients have been made, all of which are essential to construct accurate energy budgets and to convert elemental composition (i.e., carbon) to energy equivalents. Additionally, models of energetics such as the metabolic theory of ecology or dynamic energy budgets are being applied to coral physiology and include several assumptions that are not appropriate for scleractinian corals. As we assess the independent and interactive effects of multiple stressors on corals, efforts to construct quantitative energetic budgets should be a priority component of realistic multifactor experiments that would then improve the use of models as predictors of outcomes related to the effects of environmental change on corals.     

Floristic changes of epiphytic flora in the Metropolitan Lisbon area between 1980–1981 and 2010–2011 related to urban air quality

Epiphytic lichen and bryophyte Floristic Richness (FR) and distribution were investigated in Lisbon and the adjacent southern riverbank, in the centre-west of Portugal within the Lisbon Metropolitan Area (LMA). Field studies were carried out in the years 2010–2011 to replicate research conducted 30 years ago in 1980–1981.Compared to previous surveys, we confirm that the overall environmental condition has largely improved, with higher epiphyte richness. The two areas have been recolonized during the last 30 years by sensitive species mainly due to changes in SO₂ levels. However, the traffic-related NO₂ and dust deposition have become the main pollutants and the increase of nitrophilous and saxicolous/terricolous taxa reflects this influence. But, besides air pollution, the important variable affecting the epiphytic flora of LMA, currently updated to more than 200 taxa, is the influence of arborisation system type, road type and road proximity of the new surveys, in addition to urbanization (calculated in an Index of Human Impact – IHI).According to their current epiphyte diversity, with Floristic Richness (RF) ranging from 4 to more than 90 taxa (lichens and bryophytes), five zones were identified in LMA and related with air quality.As a conclusion, significant changes in the Floristic Richness (FR) were observed over the past 30 years, not only the value but also the spatial pattern which differs greatly between the two areas, linked significantly with air quality and other human influences.Due to the few number of available air quality monitoring stations, in particularly for NO₂ values, the important contributions of epiphytic flora in defining the distribution range and spatial patterns of urban disturbance imply that FR may be a practical and useful indicator of air quality in LMA.     

Seasonal effects on the sleep–wake cycle,the rest–activity rhythm and quality of life for Japanese and Thai older people

Background: The sleep-wake cycle and the rest–activity rhythm are known to change with aging, and such changes have been implicated in higher levels of depression as well as an increased incidence of dementia. However, information supporting seasonal changes in the sleep–wake cycle, the rest–activity rhythm and quality of life in older community-dwelling people remains insufficient. The aim of the present study was to prospectively investigate seasonal effects on the sleep–wake cycle, the rest–activity rhythm and quality of life among older people living in areas of Japan or Thailand with different climate classifications.

Method: The survey was conducted from March 2016 to May 2017, and 109 participants were recruited from Japan and Thailand: 47 older people living in Akita prefecture, Japan, and 62 older people living in Chiang Mai or Nakhon Ratchasima, Thailand. According to the Köppen–Geiger classification of Asian climates comprising tropical, desert, steppe, temperate and subarctic climates, Akita prefecture, which is located in northern Japan, is classified as a humid subarctic climate, while the Thai study areas are classified as tropical savanna. To monitor parameters of the sleep–wake cycle during nighttime (e.g. total sleep time, sleep latency, sleep efficiency, awaking time and frequency of sleep interruptions) and to calculate parameters of the rest–activity rhythm over the 24 h profile (e.g., interdaily stability, intradaily variability, relative amplitude, mean of least active 5 h period and mean of most active 10 h period), all the participants from both countries wore an Actiwatch 2 device on their nondominant wrist continuously for 7 days during each local season. The World Health Organization Quality of Life Questionnaire-BREF (WHOQOL-BREF) was also assessed during each local season.

Results: The final sample size was 37 older people living in Akita prefecture, Japan, and 44 older people living in Thailand; these subjects completed the data collections during each local season. The dropout rates were 21% in Japan and 29% in Thailand. The results for the Japanese subjects showed a significantly shorter sleep time with higher levels of activity during the nighttime on summer (p < 0.001) and a fragmented rest–activity rhythm over the 24 h profile on winter (p < 0.001). The older Thai participants exhibited a poor state of night sleeping year-round, and a significant relationship was observed between seasonal variations in motor activity and the social domain of WHOQOL-BREF for each Thai season (|r| = 0.4, p < 0.01).

Conclusion: These findings provide new and important information regarding seasonal effects on the sleep–wake cycle, the rest–activity rhythm and quality of life in older community-dwelling people living in two different Asian climates. Consequently, clinical preventions targeting such seasonal variations might be useful for improving the quality of life of older Japanese and Thai individuals.     

In vitro and in silico approaches to quantify the effects of the Mitraclip® system on mitral valve function

Mitraclip^® implantation is widely used as a valid alternative to conventional open-chest surgery in high-risk patients with severe mitral valve (MV) regurgitation. Although effective in reducing mitral regurgitation (MR) in the majority of cases, the clip implantation produces a double-orifice area that can result in altered MV biomechanics, particularly in term of hemodynamics and mechanical stress distribution on the leaflets.In this scenario, we combined the consistency of in vitro experimental platforms with the versatility of numerical simulations to investigate clip impact on MV functioning. The fluid dynamic determinants of the procedure were experimentally investigated under different working conditions (from 40 bpm to 100 bpm of simulated heart rate) on six swine hearts; subsequently, fluid dynamic data served as realistic boundary conditions in a computational framework able to quantitatively assess the post-procedural MV biomechanics. The finite element model of a human mitral valve featuring an isolated posterior leaflet prolapse was reconstructed from cardiac magnetic resonance. A complete as well as a marginal, sub-optimal grasping of the leaflets were finally simulated.The clipping procedure resulted in a properly coapting valve from the geometrical perspective in all the simulated configurations. Symmetrical complete grasping resulted in symmetrical distribution of the mechanical stress, while uncomplete asymmetrical grasping resulted in higher stress distribution, particularly on the prolapsing leaflet.This work pinpointed that the mechanical stress distribution following the clipping procedure is dependent on the cardiac hemodynamics and has a correlation with the proper execution of the grasping procedure, requiring accurate evaluation prior to clip delivery.     

The effects of geographical and prolonged cultural isolation on the marital behaviour of an Alpine community (Valsesia-Italy, 1618–1899)

The settlement of the “Alta Valsesia” during the XIII century by the ethnic group “Walser” coming from the Switzerland region “Vallese”, left clear historical and cultural traces. In the present study we analysed the “Alagna Valsesia” community (Piedmont). The aim of this research is to reconstruct the demographic and the matrimonial structure of this cultural and geographic isolate over three centuries and to evaluate the degree of isolation through the analysis of some biodemographic parameters calculated on the bases of parish registers, from 1618 to 1899 (1503 marriages). Data on the population dynamics highlighted an early depopulation, since 1500, than led in 1800 to a decline to population of about 60%, relating to the well known phenomenon of the leaving of mountainous isolate. The analysis of the marriages shows a low rate of endogamic mating (82%). This rate reach the highest peak 93% between 1650 and 1659, then it decreases to the minimum values 67% in the middle of 1700. The rates of consanguineous and isonymic marriages are around 20% and 4–5% respectively, reaching the maximum values during 1800; with the only exception for the period between 1700 and 1749, when the rates decreases because of a substantial migratory flow. Inbreeding coefficient stressed out — values between 0,4×10–3 and 2,4×10–3, not so different from the ones reported for other Italian mountainous isolates. The analysis of the seasonality of marriages high-lighted a strong concentration of marriages during winter months, particularly in January and February, when works were suspended because of the cold.     

A comparison of the effects of γ-ketotriazole,MCPA and carbaryl on the plasma membrane Mg2+-ATPase activity and lipid composition of rice shoots (Oryza sativa)

The effect of -ketotriazole, MCPA and carbaryl on the lipid composition and ATPase activity associated with plasma membrane fractions from rice (Oryza sativa cv Bahia) shoots was investigated. With -ketotriazole and MCPA treatments the relative amount of ⁵-avenasterol (%) was reduced in the plasma membrane, whereas with -ketotriazole a reduction was also found in the sitosterol content, expressed as a percentage of the total free sterol composition. The fatty acyl chain length of phosphatidylcholine fractions from MCPA-treated plants was also reduced. The plasma membrane Mg²⁺-ATPase activity was only stimulated in MCPA-treated plants. No changes were observed in lipid composition or ATPase activity with carbaryl treatment.     

Estimates and comparisons of the effects of sampling variation using ‘national’ macroinvertebrate sampling protocols on the precision of metrics used to assess ecological status

The Water Framework Directive (WFD) of the European Union requires all member countries to provide information on the level of confidence and precision of results in their river monitoring programmes to assess the ecological status class of river sites. As part of the European Union project STAR, the overall effects of sampling variation for a wide range of commonly used metrics and sampling methods were assessed. Replicate samples were taken in each of two seasons at 2–6 sites of varying ecological status class within each of 18 stream types spread over 12 countries, using both the STAR-AQEM method and a national sampling method or, where unavailable, the RIVPACS sampling protocol. The sampling precision of a combination of sampling method and metric was estimated by expressing the replicate sampling variance as a percentage P_samp of the total variance in metric values with a stream type; low values of P_samp indicate high precision. Most metrics had percentage sampling variances less than 20% for all or most stream types and methods. Most national methods including RIVPACS had sampling precisions at least as good as those for the STAR-AQEM method as used in their country at the same sites; the main exceptions were the national methods used in Latvia and Sweden. The national methods used in the Czech Republic, Denmark, France, Poland and the RIVPACS method used in the UK and Austria all had percentage sampling variances of less than 10% for the majority of metrics assessed. In contrast, none of the metrics had percentage sampling variances less than 10% when based on either the Italian (IBE) method, which used bank-side sorting, or the Latvian national method which identifies only a limited set of taxa. P_samp was lowest on average for the two stream types sampled in the Czech Republic using either the PERLA national method or the STAR-AQEM method. Averaged over all stream types and methods, the three Saprobic-based metrics had the lowest average percentage sampling variances (3–6%) amongst the 26 metrics assessed. These estimates of sampling standard deviation can be used to help assess the uncertainty in single or multi-metric systems for estimating site ecological status using the general STAR Bioassessment Guidance Software (STARBUGS) developed within the STAR project.     

Influence of supplemental high molecular weight pullulan or γ-cyclodextrin on ileal and total tract nutrient digestibility,fecal characteristics,and microbial populations in the dog

Abstract

This study was conducted to determine if supplemental pullulan and γ-cyclodextrin affect canine nutrient digestibility, microbial populations, and fecal characteristics. Ileal cannulated dogs were fed a commercial diet, and treatments were administered daily in a 5×5 Latin square design: (i) no supplement; (ii) 2 g pullulan; (iii) 4 g pullulan; (iv) 2 g γ-cyclodextrin; (v) 4 g γ-cyclodextrin. Ileal and fecal samples were collected the last 4 d of each 14-d period. Increasing pullulan tended (p < 0.10) to linearly increase ileal bifidobacteria and lactobacilli and quadratically increase fecal lactobacilli. A similar response was noted in ileal bifidobacteria and lactobacilli with γ-cyclodextrin. γ-Cyclodextrin resulted in a quadratic decrease (p < 0.05) in fecal Clostridium perfringens. Increasing pullulan linearly increased (p < 0.05) fecal score, while γ-cyclodextrin resulted in a linear decrease (p < 0.05). Pullulan and γ-cyclodextrin supplementation may have beneficial effects on the microbial ecology of dogs.     

Sex-specific effects of 17β-estradiol and dihydrotestosterone (DHT) on growth plate chondrocytes are dependent on both ERα and ERβ and require palmitoylation to translocate the receptors to the plasma membrane

Rat costochondral cartilage growth plate chondrocytes exhibit cell sex-specific responses to 17β-estradiol (E₂), testosterone, and dihydrotestosterone (DHT). Mechanistically, E₂ and DHT stimulate proliferation and extracellular matrix synthesis in chondrocytes from female and male rats, respectively, by signaling through protein kinase C (PKC) and phospholipase C (PLC). Estrogen receptors (ERα; ERβ) and androgen receptors (ARs) are present in both male and female cells, but it is not known whether they interact to elicit sex-specific signaling. We used specific agonists and antagonists of these receptors to examine the relative contributions of ERs and ARs in membrane-mediated E₂ signaling in female chondrocytes and DHT signaling in male chondrocytes. PKC activity in female chondrocytes was stimulated by agonists of ERα and ERβ and required intact caveolae; PKC activity was inhibited by the E₂ enantiomer and by an inhibitor of ERβ. Western blots of cell lysates co-immunoprecipitated for ERα suggested the formation of a complex containing both ERα and ERß with E₂ treatment. DHT and DHT agonists activated PKC in male cells, while AR inhibition blocked the stimulatory effect of DHT on PKC. Inhibition of ERα and ERβ also blocked PKC activation by DHT. Western blots of whole-cell lysates, plasma membranes, and caveolae indicated the translocation of AR to the plasma membrane and specifically to caveolae with DHT treatment. These results suggest that E₂ and DHT promote chondrocyte differentiation via the ability of ARs and ERs to form a complex. The results also indicate that intact caveolae and palmitoylation of the membrane receptor(s) or membrane receptor complex containing ERα and ERβ is required for E₂ and DHT membrane-associated PKC activity in costochondral cartilage cells.     

Mitochondrial DNA deletions in the peripheral blood of workers at the Mayak PA who were exposed to long-term combined effects of external γ- and internal α-radiation

The levels of large deletions in the mitochondrial DNA of workers at the Mayak Production Association (Mayak PA) who were exposed to external and combined occupational (external γ- and internal α-rays) radiation during the course of their duties were investigated. Peripheral blood-derived DNA samples were provided by the Radiobiological Human Tissue Repository of the Southern Urals Biophysics Institute (Russia). The samples were analyzed using long-extension PCR. The number of large-scale deletions in the mitochondrial DNA of workers who, in addition to external γ-radiation, were exposed to extra doses of irradiation due to incorporated ²³⁹Pu with a Pu body burden of 0.77–4.32 kBq, was 2.5-times lower compared to that of individuals who received only external γ-radiation. No significant gender-associated effects on the number of mitochondrial DNA deletions were detected among age-matched individuals.

     

Light effects on α-amylase activity and carbohydrate content in relation to lipid mobilization during the seedling growth of sunflower

The changes in α-amylase activity and in starch and free sugar content were investigated in correlation with lipid mobilization inHelianthus annuus during the first 15 days of seedling growth in discontinuous light and in darkness. Throughout the seedling development α-amylase activity increased more significantly in light than in darkness. It was always lower in cotyledons than in other tissues of the embryo axis. In both culture conditions, most of the transitory carbohydrates accumulated in germinating cotyledons were very likely synthesized by gluconeogenesis from the stored lipid breakdown. Nevertheless, in light-grown cotyledons, photosynthesis contributes to increase the carbohydrate levels. The study of several soluble sugars indicates that 1) sucrose stored in cotyledons of mature seeds was used at the onset of seedling growth, more rapidly in light than in darkness, 2) galactose and xylose, both involved as precursors of some cell-wall polysaccharides, remained at a very low level throughout the 15 days and 3) glucose, fructose and maltose accumulated in old etiolated cotyledons in contrast to what occurred in the light.