Month: January 2020

Differential expression profiles across the blood but not the sucrose feeding conditions. The most significant differences were observed between the early blood and early sucrose transcriptomes, where 5,712 genes, 35% of the total number shared, were differentially expressed. In contrast, fewer differences were seen between the late blood and late sucrose transcriptomes. The comparison of the LB, LS and teneral transcriptomes showed only 68 differentially expressed genes, which implies that 36 h after feeding on either diet, a large part of the expression profile is returning to a condition similar to that seen in non-feeding midges. The genes with differential expression profiles identified during the course of both the blood and sucrose meals were categorized according to GO classifications to depict global differences between blood and sucrose feeding. The physiological response to blood feeding was strikingly different from the teneral state, while sucrose feeding showed fewer differences in response; this stark contrast between the two diet sources was evident in both in the diversity of categories and the number of genes differentially expressed in those feeding conditions. Genes with differential expression profiles in response to blood feeding were further classified into functional categories in order to observe trends in molecular function. In comparison to the teneral midges, the early blood fed midges showed the strongest genetic response through transcript abundance changes in genes classified within anatomical structure development, cellular differentiation, reproduction, signal transduction, response to stress, transport, embryo development, lipid and ion binding, and carbohydrate metabolism. Table S4 in File S1 is a list of the top 100 genes with at least a 2-fold increase or decrease in transcript abundance as a result of an early blood meal. Intriguingly, 36 of the top 100 induced genes had no homology to other organisms, revealing new targets for functional classification. Not surprisingly, many genes that were differentially expressed in early blood feeding are associated with blood digestion, hematophagy, and vitellogenesis. At 36 h post blood feeding, we observed a general decrease in the number of genes with differential expression profiles, suggesting that many major metabolic and biological events unfold within 12 h post blood meal and the expression of these early response genes subsides within the window of 36 h. Comparison of the early and late blood-fed midge transcriptomes revealed 7,334 differentially expressed genes categorized within anatomical structure and cell differentiation, transport, ion binding, oxidoreductase, signal transduction, embryo development, response to stress, and peptidase activity, were more significantly upregulated in comparison to the measurement at early blood feeding, while categories such as reproduction, DNA binding, and cell cycle were downregulated.

FkB2 and REL were found exclusively in stromal cells, and maintained this distribution after castration. In contrast, the expression of RelB in the stroma was suppressed in response to androgen deprivation. Figure S1 summarizes these findings. By comparing the gene expression profiles in the prostate under different hormonal conditions, we identified seven TF, which were further characterized with respect to their expression in the prostate and possible variations in response to castration. These TF were found to be differentially expressed and were grouped in three functionally important groups: coordinators of the immaturity state or regulators of epithelial differentiation, coordinators of the ability of the organ to function as an immune barrier, or early-response genes or genes related to the response to extracellular signaling, which together coordinate the epithelial and/ or stromal cell behavior under androgen-deprived conditions. It is well understood that treatment with androgens increases the AR and coactivator occupancy in the PSA enhancer and promoter regions. Although the regulation of genes such as PSA by the AR in prostate cells seems to be relatively simple, and the prostate weight loss after castration is a direct effect, complex mechanisms are involved in the adaptation of the gland to varying levels of androgen stimulation. We considered that extracting information from the mass of genes revealed by the present experiments using the usual tools could be misleading, in the sense that some regulatory molecules might show subtle or no variations in mRNA content and might escape detection in assays of differential expression. Therefore, we defined an experimental approach to filter for druggable TF that could function as state-defining regulatory hubs by orchestrating different functions related to tissue remodeling and as regulators of differentiation, among others, and localizing these TF to either the epithelium and/or the stroma, and employed a MST to determine the correlation among their expression levels is a number of tissues. We identified TF genes in regulatory networks established from the enrichment terms and from the analysis of the proximal promoter region of the genes that were regulated by androgen deprivation, as revealed by the microarray analysis and referenced not only to the normal androgen levels in non-castrated rats, but also to the high-dose estrogen stimulus. The search for biological relationship between the TF found in the selected networks and the steroid hormone receptors Ar, Esr1 and Esr2 was unveiled by calculating their expression correlation and constructing a MST. MYBL2 is related to proliferative activity. Use of the MMTVPyMT mouse was instrumental in the discovery that Myb is essential for the development of mammary tumors, but not ultimately for their progression.

Many of the genes that are essential for the functioning of the vertebrate HPI axis were not expressed in 3dpf stickleback embryos, including several key neuroendocrine ligands, their receptors, and enzymes responsible for the production of cortisol. Also, we detected a significant effect of maternal stress on the expression of genes shown to be similarly influenced in humans and other mammal cells by exposure to the synthetic glucocorticoid dexamethasone. It is therefore possible that some of the genes differentially expressed in stickleback embryos in response to maternal stress were activated or repressed through the action of maternal cortisol, though cortisol was not measured in this experiment. Further study is needed to understand the magnitude and tissue specificity by which maternal glucocorticoids might influence the embryonic transcriptome. Consistent with the known effects of glucocorticoids on immune function, we detected the differential regulation of several immune genes, including genes involved in the innate and adaptive immune responses. Previous studies in humans and other primates have found an enhancing effect of maternal stress on the offspring innate immune response, coupled with a depression of the adaptive immune response. Cardiovascular disease is a major cause of death worldwide, especially in developed countries. Higher concentrations of circulating cholesterol are a major risk factor for cardiac problems. Limiting cholesterol intake to 300 mg/day can prevent elevated blood cholesterol levels and prevent increases in the risk of coronary heart diseases. This figure is given in both the National Cholesterol Education Program diet and the American Heart Association dietary recommendations. In developed countries, egg consumption has decreased considerably because the cholesterol, 200–300 mg per egg, can increase the risk of coronary heart disease. However, in China and or other developing countries, people usually eat eggs as their main source of dietary protein. Low-cholesterol eggs would be of great significance to public health. Ever since the 1970s, research efforts have been directed toward reducing egg cholesterol content in different ways. These have included genetic selection, altering the hens’ diet with various nutrients and probiotics, and treating the hens with drugs, non-nutritive factors, and phytogenic extracts. Consumers are likely to prefer eggs modified by phytogenic extract to eggs modified by drugs because they have fewer and less side effects. Alfalfa saponin extract is extracted from Medicago sativa L. The main active component is saponins. It also includes flavonoids and polysaccharides. The mixture of biologically active ingredients presented in ASE has pleiotropic physiological effects, including reduction of aphid population, alteration nutrient digestion and fermentation in rumen, antimicrobial activity.

Proteins are transported via the cell membrane and can catalyze reactions, and many proteins form aggregates and amyloidal structures in conditions when they pass to intermediate states. Herein we have tried to clarify which amino acid residues determine the stability of the molten globule state of apomyoglobin. On the one hand, it can be proposed that the formation of this intermediate state is affected by hydrophobic amino acid residues in the protein hydrophobic core since it is known that at the first stage of folding hydrophobic collapse of the polypeptide chain occurs. On the other hand, we can suggest that the hydrophobic amino acid residues on the protein surface interacting with the solvent molecules also affect the molten globule state. The substitution of hydrophobic residues on the protein surface can have an effect on protein misfolding, i.e. decrease or increase the probability of formation of irregular hydrophobic contacts during protein folding. We have also postulated that the introduced disulfide bond can influence the molten globule state because the SS-bond should affect the mobility and compactness of the protein. By our hypothesis, the disulfide bond should be introduced on the protein surface so that the packing of the hydrophobic core of apomyoglobin would remain undamaged. To verify the above proposals, we have studied four mutant forms of apomyoglobin with substitutions of hydrophobic amino acid residues on its surface and ten mutant forms of apomyoglobin with substitutions of amino acid residues in the hydrophobic core of this protein. In addition, we have analyzed the mutant form of apomyoglobin with the introduced disulfide bond on the protein surface. They are large hydrophobic amino acid residues located in different structural elements of apomyoglobin. Earlier we investigated the effect of these residues on the rates of refolding and unfolding of the apomyoglobin structure. In this study, we analyze only the influence of single substitutions of hydrophobic amino acid residues on the stability of the molten globule state of apomyoglobin. Hydrophobic amino acid residues on the protein surface were also chosen for substitutions. Based on the crystal structure of myoglobin we selected residues that are maximally exposed to the aqueous environment and weakly interact with other amino acid residues of the protein. Six hydrophobic amino acid residues were selected to be substituted by hydrophilic ones. Four mutant proteins were examined: with substitutions of two, three, four and six amino acid residues. Later the mutant proteins were designated as m2, m3, m4, and m6, respectively. Fig. 1 shows a three-dimensional model of apomyoglobin. As can be seen, the chosen amino acid residues on the surface of apomyoglobin are grouped on one side of the protein. We suggest that such positions of residues can enhance the effect of mutation on the molten globule state of apomyoglobin.

Observation of an absence of a bystander effect in the present study may help explain the sensitivity of radioresistant tumor cells to neutrons, because there is a possibility that the protection otherwise provided by the bystander effect on the tumor in response to neutrons is absent or not strong enough in magnitude, thereby causing tumor cells to be killed. The risks currently associated with neutron exposure may be over or underestimated depending on which model of risk estimation is used to predict low dose risks from high dose data. Hence, reevaluation of radiation protection standards may be required. The work described in this paper may be relevant for radiation oncologists planning cancer treatments that involve fast neutron or proton radiotherapy, particularly for pediatric patients or pregnant women. This study used cells that lack gap junctions. There is a possibility that a neutron-induced bystander effect requires physical contact between cells, which could be tested by performing experiments using cell lines such as fibroblasts and keratinocytes that have gap junctions. If no bystander effect is induced in these cell lines, then it may be likely that neutrons do not have any ability to induce a bystander effect. Another possible explanation for the lack of a bystander effect with neutrons observed in this study may be the presence of dimethyl sulfoxide, a scavenger of reactive oxygen species, which was used to dissolve cytochalasin B that is required for the cytokinesis-block micronucleus assay. Both pre- and post-radiation treatment with DMSO is known to suppress DNA damage in irradiated cells. However, this possibility seems unlikely in the work described here because we observed a bystander effect with an identical procedure involving DMSO when the same cell lines were exposed to photons. However, if a very small bystander effect was in fact induced by neutrons, then it may have been obscured by the DMSO, whereas the bystander effect induced by high levels of photons was too large to be masked by DMSO. For cells irradiated with high doses of photons, a considerable amount of damage was attributed to the bystander component. The percent contribution by the bystander exposure to the direct exposure was highest at the lowest dose delivered and then it appears to saturate as dose increases, perhaps because there is saturation either of the bystander signals or the cellular responses to those signals. This observation is in agreement with other reports. For cells irradiated with neutrons there is little or no damage that can be attributed to a bystander effect, because as previously noted, there is comparatively less oxidative damage following neutron than gamma exposure. We report two RBEs for neutron radiation, one for micronuclei and the other for nucleoplasmic bridges. These two genetic endpoints have different mechanisms of formation.