The Effect of Dietary β-Sitosterol and β-Sitostanol on the Metabolism of Cholesterol in Rats

Effects of dietary β-sitosterol (S) and β-sitostanol (HS) on the metabolism and fate of labeled cholesterol intravenously injected were compared in rats fed diets high in cholesterol. Kinetic behavior of the decay curve for serum cholesterol in the HS supplemented (C + HS) group approximated to that in the cholesterol-free (control) group. The largest dilution of the label was observed in rats of the cholesterol (C) group and the least in the C + HS group, the C + S group being intermediate. The specific activity of hepatic cholesterol was in the decreasing order of the C + HS, C + S and C groups, while the situation was reversed when expressed in terms of net incorporation. Thus, cholesterol pool seemed to be much smaller in the C + HS group than in the C + S group.

In a long term feeding experiment with diets free of cholesterol, HS exhibited significantly greater hypocholesterolemic activity than S did.

These data, together with those reported previously, indicated that inhibitory effect on the absorption of both endogenous and exogenous cholesterol was much more greater in HS than in S.     

Early Training-Induced Reduction of Angiotensinogen in Autonomic Areas—The Main Effect of Exercise on Brain Renin-Angiotensin System in Hypertensive Rats

Background

Exercise training (T) blunts functional deficits and renin-angiotensin system (RAS) hyperactivity in hypertensive individuals. There is no information on T-induced temporal changes of brain RAS. We evaluate now the simultaneous effects of T on functional responses and time course changes in the expression/activity of brain RAS components in autonomic cardiovascular-controlling areas.

Methods and Results

Spontaneously hypertensive rats (SHR) and age-matched normotensive controls (WKY) were trained for 0, 1, 2, 4, 8 and 12 weeks. Sedentary (S) groups served as time-controls. After arterial pressure (AP) and heart rate (HR) recordings at rest, fresh and fixed brains were harvested for qPCR and immunofluorescence assays. SHR-S vs. WKY-S exhibited higher mean AP (MAP) and HR, increased pressure variability and sympathetic activity, elevated AT₁ receptor (AT₁) expression in nucleus tractus solitarii (NTS) and higher Mas receptor expression in the rostroventrolateral medulla (RVLM). In SHR, T promptly (T₂ on) reduced sympathetic variability to heart/vessels and largely decreased angiotensinogen expression in the paraventricular hypothalamic nucleus (PVN) and NTS, with a late RVLM reduction (T₄). AT₁ expression was only reduced at T₁₂ (PVN and NTS) with transient, not maintained Mas receptor changes in PVN and RVLM. These responses were accompanied by baseline MAP and HR reduction in the SHR-T (from T₄ on). In the SHR group, PVN angiotensinogen expression correlated positively with sympathetic activity, resting MAP and HR. In WKY-T, a precocious (T₂-T₁₂) RVLM AT₁ decrease preceded the appearance of resting bradycardia (from T₈ on).

Conclusions

Early and maintained reduction of angiotensinogen content in autonomic areas of the SHR is the most prominent effect of training on brain RAS. Down-regulation of PVN RAS expression is an essential factor to drive cardiovascular benefits in SHR-T, while resting bradycardia in WKY-T is correlated to RVLM AT₁ reduction.     

The Effect of Elevated Concentrations of Fructose 2,6-Bisphosphate on Carbon Metabolism during Deacidification in the Crassulacean Acid Metabolism Plant Kalanchöe daigremontiana

In C₃ plants, the metabolite fructose 2,6-bisphosphate (Fru 2,6-P₂) has an important role in the regulation of carbon partitioning during photosynthesis. To investigate the impact of Fru 2,6-P₂ on carbon metabolism during Crassulacean acid metabolism (CAM), we have developed an Agrobacterium tumefaciens-mediated transformation system in order to alter genetically the obligate CAM plant Kalanchöe daigremontiana. To our knowledge, this is the first report to use genetic manipulation of a CAM species to increase our understanding of this important form of plant metabolism. Transgenic plants were generated containing a modified rat liver 6-phosphofructo-2-kinase gene. In the plants analyzed the activity of 6-phosphofructo-2-kinase ranged from 175% to 198% of that observed in wild-type plants, resulting in Fru 2,6-P₂ concentrations that were 228% to 350% of wild-type plants after 2 h of illumination. A range of metabolic measurements were made on these transgenic plants to investigate the possible roles of Fru 2,6-P₂ during Suc, starch, and malic acid metabolism across the deacidification period of CAM. The results suggest that Fru 2,6-P₂ plays a major role in regulating partitioning between Suc and starch synthesis during photosynthesis. However, alterations in Fru 2,6-P₂ levels had little effect on malate mobilization during CAM fluxes.     

Effect of 2-deoxy-d-Glucose on Aminoacids Metabolism in Rats’ Cerebral Cortex Slices

We studied the effect of different concentrations of 2-deoxy-d-glucose on the l-[U-¹⁴C]leucine, l-[1-¹⁴C]leucine and [1-¹⁴C]glycine metabolism in slices of cerebral cortex of 10-day-old rats. 2-deoxy-d-glucose since 0.5 mM concentration has inhibited significantly the protein synthesis from l-[U-¹⁴C]leucine and from [1-¹⁴C]glycine in relation to the medium containing only Krebs Ringer bicarbonate. Potassium 8.0 mM in incubation medium did not stimulate the protein synthesis compared to the medium containing 2.7 mM, and at 50 mM diminishes more than 2.5 times the protein synthesis compared to the other concentration. Only at the concentration of 5.0 mM, 2-deoxy-d-glucose inhibited the CO₂ production and lipid synthesis from l-[U-¹⁴C] leucine. This compound did not inhibit either CO₂ production, or lipid synthesis from [1-¹⁴C]glycine. Lactate at 10 mM and glucose 5.0 mM did not revert the inhibitory effect of 2-deoxy-d-glucose on the protein synthesis from l-[U-¹⁴C]leucine. 2-deoxy-d-glucose at 2.0 mM did not show any effect either on CO₂ production, or on lipid synthesis from l-[U-¹⁴C]lactate 10 mM and glucose 5.0 mM.     

Effect of Sodium Nitrite on Ischaemia and Reperfusion-Induced Arrhythmias in Anaesthetized Dogs: Is Protein S-Nitrosylation Involved?

Background and Purpose

To provide evidence for the protective role of inorganic nitrite against acute ischaemia and reperfusion-induced ventricular arrhythmias in a large animal model.

Experimental Approach

Dogs, anaesthetized with chloralose and urethane, were administered intravenously with sodium nitrite (0.2 µmolkg^-1min^-1) in two protocols. In protocol 1 nitrite was infused 10 min prior to and during a 25 min occlusion of the left anterior descending (LAD) coronary artery (NaNO₂-PO; n = 14), whereas in protocol 2 the infusion was started 10 min prior to reperfusion of the occluded vessel (NaNO₂-PR; n = 12). Control dogs (n = 15) were infused with saline and subjected to the same period of ischaemia and reperfusion. Severities of ischaemia and ventricular arrhythmias, as well as changes in plasma nitrate/nitrite (NOx) levels in the coronary sinus blood, were assessed throughout the experiment. Myocardial superoxide and nitrotyrosine (NT) levels were determined during reperfusion. Changes in protein S-nitrosylation (SNO) and S-glutathionylation were also examined.

Key Results

Compared with controls, sodium nitrite administered either pre-occlusion or pre-reperfusion markedly suppressed the number and severity of ventricular arrhythmias during occlusion and increased survival (0% vs. 50 and 92%) upon reperfusion. There were also significant decreases in superoxide and NT levels in the nitrite treated dogs. Compared with controls, increased SNO was found only in NaNO₂-PR dogs, whereas S-glutathionylation occurred primarily in NaNO₂-PO dogs.

Conclusions

Intravenous infusion of nitrite profoundly reduced the severity of ventricular arrhythmias resulting from acute ischaemia and reperfusion in anaesthetized dogs. This effect, among several others, may result from an NO-mediated reduction in oxidative stress, perhaps through protein SNO and/or S-glutathionylation.     

Effect of Seawater–Sewage Cross-Transplants on Bacterial Metabolism and Diversity

Bioassays experiments were conducted to determine the metabolic and community composition response of bacteria to transplants between relatively pristine coastal seawater and sewage-impacted seawater. There were four treatments: (1) pristine seawater bacteria?+?pristine seawater (Pb?+?Pw), (2) sewage-impacted bacteria?+?sewage-impacted water (Sb?+?Sw), (3) pristine seawater bacteria?+?sewage-impacted water (Pb?+?Sw), and (4) sewage-impacted bacteria?+?pristine seawater (Sb?+?Pw). Sewage-derived DOC was more labile and readily utilized by bacteria, which favored the growth of high nucleic acid (HNA) bacteria, resulting in high bacterial production (BP, 113?±?4.92 to 130?±?15.8 μg C l^?1?day^?1) and low respiration rate (BR, <67?±?11.3 μg C l^?1?day^?1), as well as high bacterial growth efficiency (BGE, 0.68?±?0.09 to 0.71?±?0.05). In contrast, at the relatively pristine site, bacteria utilized natural marine-derived dissolved organic matter (DOM) at the expense of lowering their growth efficiency (BGE, <0.32?±?0.02) with low BP (<62?±?6.3 μg C l^?1?day^?1) and high BR 133?±?14.2 μg C l^?1?day^?1). Sewage DOM input appeared to alter the partitioning of carbon between respiration and production of bacteria, resulting in a shift toward higher BGE, which would not enhance oxygen consumption. Taxonomic classification based on 454 pyrosequencing reads of the 16S rRNA gene amplicons revealed that changes in bacterial community structure occurred when seawater bacteria were transferred to the eutrophic sewage-impacted water. Sewage DOM fueled the growth of Gammma-proteobacteria and Epsilson-proteobacteria and reduced the bacterial richness, but the changes in the community were not apparent when sewage-impacted bacteria were transferred to pristine seawater.     

The coming of age of niche vaccines?: Effect of vaccines on resistance profiles in Streptococcus pneumoniae

Pneumococcal conjugate vaccine is not only effective against invasive and non-invasive disease in all ages but also has a significant impact on antibiotic-resistant infections since a number of resistant serotypes are targeted by the vaccine. Surveillance studies in the post-licensure years have shown an increase in non-vaccine types both in carriage and disease isolates. There appears to be an increasing trend in antimicrobial resistance among these non-vaccine types, especially in serotype 19A. The impact these non-vaccine types will have on disease remains to be seen, but clearly continued efforts at characterizing the pneumococcal population are essential for future surveillance. Although expanded vaccine formulations with efficacy against a wider range of serotypes will be useful, reduction of drug-resistant Streptococcus pneumoniae will require a combination of the conjugate vaccine and a reduction in antimicrobial use.     

Effect of Recombinant Interferon-α2b (Laferon) on Transport of Sodium Ions in Human Neuroblastoma Cells

To elucidate molecular mechanisms of neurotropic action of a recombinant interferon, IFN-2b (laferon), its effect on transport of ²²Na⁺ through the membrane of cultured human neuroblastoma cells (line IMR 32) was investigated. Within the first minutes after treatment with IFN-2b, the influx of ²²Na⁺ ions was reduced by 20%, as compared with the control. Depolarization of the plasma membrane by a mixture of veratrine and scorpion (Leiurus quinquestriatus) toxin (200 and 10 g/ml, respectively) increased this flux by 50% in the control and by 70% in the IFN-2b-treated cells. A blocker of voltage-operated sodium channels, tetrodotoxin (TTX, 4 · 10^-7 M), suppressed the inward flux of ²²Na⁺ ions (completely in the control cells and by 75% in the IFN-2b-treated cells). The influx of ²²Na⁺ ions into neuroblastoma cells depended on the concentration of IFN-2b in the incubation medium, reaching a maximum at concentrations of 600-1000 IU/ml. This allows us to suggest that entry of Na⁺ ions into neuroblastoma cells caused by IFN-2b is basically performed through voltage-operated TTX-sensitive sodium channels.     

Effect of Selenium on Ascorbate�CGlutathione Metabolism During PEG-induced Water Deficit in Trifolium repens L.

To elucidate the effect of selenium (Se) on the ascorbate?Cglutathione (ASC?CGSH) cycle under drought stress, the activities of antioxidant enzymes and the levels of molecules involved in ASC?CGSH metabolism were studied in Trifolium repens seedlings subjected to polyethylene glycol (PEG)-induced water deficit alone or combined with 5???M Na₂SeO₄. Compared to the control, H₂O₂, thiobarbituric acid reactive substances (TBARS), ascorbate (ASC), dehydroascorbate (DHA), and glutathione disulfide (GSSG) contents increased, whereas a constant content of glutathione (GSH) and decreases in ASC/DHA and GSH/GSSG ratios were observed in the presence of PEG. The activities of ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), and glutathione reductase (GR) were upregulated, except for monodehydroascorbate reductase (MDHAR) activity during PEG-induced water deficit. Se application decreased the contents of H₂O₂, TBARS, DHA, and GSSG, increased the levels of GSH and ASC, and inhibited the decreases of ASC/DHA and GSH/GSSG ratios. Although it did not affect APX activity significantly, Se addition improved the activities of MDHAR, DHAR, and GR. Furthermore, GR activity showed the highest increase followed by that of DHAR and MDHAR in decreasing order. These data indicated that fluctuations in ASC?CGSH metabolism resulting from Se may have a positive effect on drought stress mitigation, and the regulation in the ASC?CGSH cycle can be attributed mainly to GR and DHAR in PEG?+?Se-treated T. repens seedlings.     

Showoffs or Providers? The Parenting Effort of Hadza Men

Paternal care plays an important role in many scenarios of human evolution. Lately, however, this “Man the Provisioner” view has been challenged. The showoff hypothesis, for example, proposes that men hunt not to provision children but to gain extra mating opportunities, and some have suggested that male care among mammals is always a form of mating, rather than parenting, effort. This study, based on observation in a hunting and gathering society, the Hadza of Tanzania, tests whether men provide care as parenting effort. If male care were mating effort only, stepchildren should receive no less care than biological children. My data, however, reveal that stepchildren do receive less care. This suggests that care is provided, at least in part, as parenting effort. Although lower direct care implies stepfathers are less motivated to care for stepchildren, resource acquisition data raise the question of whether stepfathers are less motivated, or simply less skilled.     

Effect of Time‐of‐Day‐Specific Strength Training on Serum Hormone Concentrations and Isometric Strength in Men

A time‐of‐day influence on the neuromuscular response to strength training has been previously reported. However, no scientific study has examined the influence of the time of day when strength training is performed on hormonal adaptations. Therefore, the primary purpose of this study was to examine the effects of time‐of‐day‐specific strength training on resting serum concentrations and diurnal patterns of testosterone (T) and cortisol (CORT) as well as maximum isometric strength of knee extensors. Thirty eight diurnally active healthy, previously untrained men (age 20–45 yrs) underwent a ten‐week preparatory strength training period when sessions were conducted between 17:00–19:00 h. Thereafter, these subjects were randomized into either a morning (n=20, training times 07:00–09:00 h) or afternoon (n=18, 7:00–19:00 h) training group for another ten‐week period of time‐of‐day‐specific training (TST). Isometric unilateral knee extension peak torque (MVC) was measured at 07:00, 12:00, 17:00, and 20:30 h over two consecutive days (Day 1 & Day 2) before and after TST. Blood samples were obtained before each clock‐time measurement to assess resting serum T and CORT concentrations. A matched control group (n=11) did not train but participated in the tests. Serum T and CORT concentrations significantly declined from 07:00 to 20:30 h on all test days (Time effect, p<.001). Serum CORT at 07:00 h was significantly higher on Day 1 than Day 2 in the control and afternoon group, both in Pre and Post conditions (Day×Time interaction, p<.01). In the morning group, a similar day‐to‐day difference was present in the Pre but not Post conditions (Time×Group interaction, p<.05). MVC significantly increased after TST in both the morning and afternoon groups (Pre to Post effect, p<.001). In both groups, a typical diurnal variation in MVC (Time effect, p<.001) was found, especially on Day 2 in the Pre condition, and this feature persisted from Pre to Post in the afternoon group. In the morning group, however, diurnal variation was reduced after TST on both Day 1 and Day 2 (Pre to Post×Day×Time×Group interaction, p<.05). In conclusion, 10 weeks of morning time‐of‐day‐specific strength training resulted in reduced morning resting CORT concentrations, presumably as a result of decreased masking effects of anticipatory psychological stress prior to the morning testing. The typical diurnal pattern of maximum isometric strength was blunted by the TST period in the morning but not the afternoon group. However, the TST period had no significant effect on the resting total T concentration and its diurnal pattern and on the absolute increase in maximum strength.     

Home alone? Effect of weather-induced behaviour on spread of SARS-CoV-2 in Germany

In early 2020 the world was struck by the epidemic of novel SARS-CoV-2 virus. Like many others, German government has introduced severe contact restrictions to limit the spread of infection. This paper analyses effects of weather on the spread of the disease under the described circumstances. We demonstrate that regions reported lower growth rates of the number of the infection cases after days with higher temperatures, no rain and low humidity. We argue that this effect is channelled through human behaviour. The evidence suggests that “good” weather attracts individuals to outdoor (safer) environments, thus, deterring people from indoor (less safe) environments. Understanding this relationship is important for improving the measures aiming at combating the spread of the virus.     

Phytoremediation in the Tropics—The Effect of Crude Oil on the Growth of Tropical Plants

Phytoremediation is a nondestructive, cost-effective in-situ technology to clean up contaminated soils. In the case of contamination with petroleum hydrocarbons, plants enhance microbial degradation of the contaminant in the rhizosphere. The potential of this technology for the tropics should be high due to prevailing climatic conditions favoring plant growth and stimulating microbial activity. Investigations of the potential of tropical plants for phytoremediation, however, are scarce. The present work studied two grasses and six legumes from the eastern savannah of Venezuela on their reaction to crude oil contamination in soil. Results shall help to identify plants with a potential for phytoremediation and subsequent studies. Seedling emergence and biomass production were determined for plants growing in soil contaminated with 0%, 3%, and 5% heavy crude oil. Contamination had, in general, a tendential but not significant negative influence on seedling emergence. Dry matter production was reduced by only a few percent to up to 85%. Furthermore, in some legumes inhibition of nodulation was observed. The grass Brachiaria brizantha and the legumes Centrosema brasilianum and Calopogonium mucunoides are promising for phytoremediation because in contaminated soil they combined high seedling emergence with least affected biomass production. Since they are cultivated forage/soil cover species also in other regions of the tropics, their potential for phytoremediation of petroleum contaminated soils extends beyond Venezuela.     

The Effect of Attachment Site Mutations on Strand Exchange in Bacteriophage λ Site-Specific Recombination

Recombination of phage λ attachment sites occurs by sequential exchange of the DNA strands at two specific locations. The first exchange produces a Holliday structure, and the second resolves it to recombinant products. Heterology for base substitution mutations in the region between the two strand exchange points (the overlap region) reduces recombination; some mutations inhibit the accumulation of Holliday structures, others inhibit their resolution to recombinant products. To see if heterology also alters the location of the strand exchange points, we determined the segregation pattern of three single and one multiple base pair substitution mutations of the overlap region in crosses with wild type sites. The mutations are known to differ in the severity of their recombination defect and in the stage of strand exchange they affect. The three single mutations behaved similarly: each segregated into both products of recombination,, and the two products of a single crossover were frequently nonreciprocal in the overlap region. In contrast, the multiple mutation preferentially segregated into one of the two recombinant products, and the two products of a single crossover appeared to be fully reciprocal. The simplest explanation of the segregation pattern of the single mutations is that strand exchanges occur at the normal locations to produce recombinants with mismatched base pairs that are frequently repaired. The segregation pattern of the multiple mutation is consistent with the view that both strand exchanges usually occur to one side of the mutant site. We suggest that the segregation pattern of a particular mutation is determined by which stage of strand exchange it inhibits and by the severity of the inhibition.