Four mouse bone marrow or thymus cell populations, namely granulopoietic/monocytopoietic, erythropoietic, B-lymphopoietic, and T-lymphopoietic precursor cells have been assayed by RTPCR technique for the presence and relative amounts of adenosine A1, A2a, A2b, and A3 receptor mRNA. It has been found that (i) all four populations studied express all four adenosine receptor subtypes, (ii) the A1 receptor is the least expressed in all populations studied, (iii) the A3 receptor is markedly expressed in the populations of granulopoietic/monocytopoietic and erythropoietic cells, (iv) the A2a receptor is markedly expressed in the populations of B-lymphopoietic and T-lymphopoietic cells, and v) the A2b receptor does not predominate in any of the precursor cells studied. Our data offer a new possibility for the assessment of the readiness of these cells to respond, by receptor-mediated mechanisms, to adenosine or its analogs present in the tissues as a result of endogenous processes and/or following their administration., D. Štreitová ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
Expression of mRNA for adenosine receptor subtypes A1, A2a, A2b, and A3 in normal and lipopolysaccharide (LPS)-activated murine RAW 264.7 macrophages has been investigated using the method of quantitative real-time polymerase chain reaction. The results have shown a very low, unquantifiable expression of adenosine A1 receptor mRNA in both normal and LPS-activated macrophages. The other three adenosine receptor mRNAs have been found to be expressed at various but always quantifiable levels. Activation of the macrophages by LPS induced upregulation of the expression of adenosine receptor A2a and A2b mRNA, whereas the expression of adenosine receptor A3 mRNA was downregulated. Unstimulated macrophages exhibited a high expression of the A2b adenosine receptor mRNA. The findings are discussed from the point of view of the antiinflammatory and hematopoiesis-stimulating roles of the adenosine receptor signaling., D. Štreitová ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
Cytochrome P450s (P450s) involved in insecticide resistance reduce the efficacy of insecticide-based vector control by rendering vector control ineffective. They are recorded in many species of vectors and have various constitutive and insecticide induction profiles. In this study, the isolation and prediction of the structure of a P450 from a strain of Aedes aegypti originating from Malaysia is reported. Quantitative mRNA expression of this gene and a previously reported P450, CYP4H28v2, in the developmental stages of the mosquito after exposure to sub-lethal concentrations of insecticides is also reported. The isolated P450, CYP4H31v2, is an allelic variant of CYP4H31 and contains several conserved motifs of P450s. The secondary structure of the protein is mostly made up of alpha helices and random coils. The tertiary structure was generated using homology modeling and was of good quality based on structure validation using protein structure assessment tools. CYP4H28v2 and CYP4H31v2 were differentially expressed in the developmental stages of the vector, with a significantly increased expression in adult males. The genes were significantly over-expressed in larvae exposed to deltamethrin and permethrin for 6 h. In the DDT-treated larvae, only CYP4H31v2 was significantly over-expressed after a 6 h exposure. Under-expression of the genes was predominant in larvae treated with the organophosphates malathion and temephos. Though the functions of these P450s are unknown, their response to induction by exposure to insecticides indicates the likely involvement of these genes in insecticide tolerance. and Fatma M. A. El-Garj, Mustafa F.F. Wajidi, Silas W. Avicor.
mRNA expression patterns of genes for metabolic key enzymes sucrose phosphate synthase (SPS), phosphoenolpyruvate carboxylase (PEPC), pyruvate kinase, ribulose 1,5-bisphosphate carboxylase/oxygenase, glutamine synthetase 1, and glutamine synthetase 2 were investigated in leaves of rice plants grown at two nitrogen (N) supplies (N0.5, N3.0). The relative gene expression patterns were similar in all leaves except for 9th leaf, in which mRNA levels were generally depressed. Though increased N supply prolonged the expression period of each mRNA, it did not affect the relative expression intensity of any mRNA in a given leaf. SPS Vmax, SPS limiting and PEPC activities, and carbon flow were examined. The ratio between PEPC activity and SPS Vmax was higher in leaves developed at the vegetative growth stage (vegetative leaves: 5th and 7th leaves) than in leaves developed after the ear primordia formation stage (reproductive leaves: 9th and flag leaves). PEPC activity and SPS Vmax decreased with declining leaf N content. After using 14CO2 the 14C photosynthate distribution in the amino acid fraction was higher in vegetative than in reproductive leaves when compared for the same leaf N status. Thus, at high PEPC/SPS activities ratio, more 14C photosynthate was distributed to the amino acid pool, whereas at higher SPS activity more 14C was channelled into the saccharide fraction. Thus, leaf ontogeny was an important factor controlling photosynthate distribution to the N- or C-pool, respectively, regardless of the leaf N status. and T. Shinano ... [et al.].
GIP (glucose dependent insulinotr ophic polypeptide), originally identified as an incretin peptide synthesized in the gut, has recently been identified, along with its receptors (GIPR), in the brain. Our objective was to investigate the role of GIP in hypothalamic gene expression of biomarkers linked to regulating energy balance and feeding behavi or related neurocircuitry. Rats with lateral cerebroventricular cannulas were administered 10 μg GIP or 10 μl artificial cerebrospinal fluid (aCSF) daily for 4 days, after which whole hypothalami were collected. Real time Taqman™ RT-PCR was used to quantitatively compare the mRNA expression levels of a set of genes in the hypothalamus. Administration of GIP resulted in up-regulation of hypothalamic mRNA levels of AVP (46.9±4.5 %), CART (25.9±2.7 %), CREB1 (38.5±4.5 %), GABRD (67.1±11 %), JAK2 (22.1±3.6 %), MAPK1 (33.8±7.8 %), NPY (25.3±5.3 %), OXT (49.1±5.1 %), STAT3 (21.6±3.8 %), and TH (33.9±8.5 %). In a second experiment the same set of genes was evaluated in GIPR -/- and GIPR +/? mice to determine the effect of lack of GIP stimulation on gene expression. In GIPR -/- mice expressions of the following genes were down-regulated: AVP (27. 1±7.5 %), CART (28.3±3.7 %), OXT (25.2±5.8 %), PTGES (23.9±4.5 %), and STAT3 (8.8±2.3 %). These results suggest that AVP, CART, OXT and STAT3 may be involved in energy balance-related hypothalamic circuits affected by GIP., S. Ambati ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy
In the present study, we have investigated the role of antimalarial drug halofantrine (HF) in inducing the sterile protection against challenges with sporozoites of the live infectious Plasmodium yoelii (Killick-Kendrick, 1967) in Swiss mice malaria model. We observed that during the first to third sequential sporozoite inoculation cycles, blood-stage patency remains the same in the control and chemoprophylaxis under HF drug cover (CPS-HF) groups. However, a delayed blood-stage infection was observed during the fourth and fifth sporozoite challenges and complete sterile protection was produced following the sixth sporozoite challenge in CPS-HF mice. We also noticed a steady decline in liver stage parasite load after 3th to 6th sporozoite challenge cycle in CPS-HF mice. CPS-HF immunisation results in a significant up-regulation of pro-inflammatory cytokines (IFN-γ, TNF-α, IL-12 and iNOS) and down-regulation of anti-inflammatory cytokines (IL-10 and TGF-β) mRNA expression in hepatic mononuclear cells (HMNC) and spleen cells in the immunised CPS-HF mice (after 6th sporozoite challenge) compared to control. Overall, our study suggests that the repetitive sporozoite inoculation under HF drug treatment develops a strong immune response that confers protection against subsequent challenges with sporozoites of P. yoelii.