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Myostatin (MSTN) and transforming growth factor-ß1 (TGF-ß1) belong to the same TGF-ß superfamily of proteins. They are involved in regulation of skeletal muscle growth and development as well as muscle catabolism. The aim of the present study was to investigate the relationship between mstn and TGF-ß1 expression in proliferating and differentiating mouse C2C12 myoblasts cultured in normal and catabolic conditions and to evaluate the effect of exogenous TGF-ß1 as well as "knock down" of TGF-ß1 receptor type II on mstn expression in proliferating and differentiating myogenic cells. The direct effect of TGF-ß1 on myostatin was also examined. Myostatin expression increased gradually with cell confluency in proliferating cultures, while the level of TGF-ß1, detected in the form of a 100 kDa small latent complex diminished. Myostatin expression was accompanied by a partial cell cycle arrest. Three forms of myostatin were found: a 52 kDa precursor, a 40 kDa latency associated propeptide, and a 26 kDa active peptide. A decrease in myostatin and TGF-ß1 levels was observed during the first three days of differentiation, which was subsequently followed by significant increase of their expression during next three to four days of differentiation. Catabolic state induced by dexamethasone significantly increased the level of all forms of myostatin as well as latent (100 kDa) and active (25 kDa) forms of TGF-ß1 in differentiating myoblasts in a dose dependent manner. Exogenous TGF-ß1 (2 ng/ml) significantly increased myostatin levels both in proliferating and differentiating C2C12 myoblasts, whereas silencing of the TGF-ß1 receptor II gene significantly lowered myostatin level in examined cells. The presented results indicate that TGF-ß1 may control myostatin-related regulation of myogenesis through up-regulation of myostatin, predominantly in the course of terminal differentiation and glucocorticoid-dependent catabolic stimulation.
The therapeutic effect of 1 mg/kg of dexamethasone on lung oedema and respiratory distress induced by 0.08 mg/kg of oleic acid (OA) in dogs was investigated into 8 dogs. One hour after OA injection, blood pH, HCO₃, pO₂, O₂Hb, and heart rate decreased, whereas COHb, pCO₂, temperature, and respiration rate increased. Lung oedema was confirmed histopathologically in a dog from the control group who died. Blood pH, COHb, temperature, and respiration rate returned to normal values after administration of dexamethasone. It has been determined that respiratory distress and lung oedema cause important changes in blood gases and the administration of a single dose of 1 mg/kg of dexamethasone is effective in the treatment of lung oedema and respiratory distress induced in dogs by OA.
The aim of the study was to examine the changes in the density of VAChT (marker of acetylcholine present)-, NPY-, VIP-, SOM-, SP- and nNOS-immunoreactive (IR) nerve terminals and co-localization of VAChT with the above-mentioned neurotransmitters after the occurrence of dexamethasone (DXM)-induced ovarian cysts in gilts. DXM administration led to an increase in the density of VAChT/SP-, VAChT/nNOS- and NPY-IR nerve terminals around the cystic walls. In DXM-treated animals an elevated number of VAChT- and SP-IR nerve endings was found close to the tertiary follicles. Moreover, in the gilts receiving DXM the density of NPY-IR nerve endings (that simultaneously co-localized VAChT) was high near the interstitial gland. An increase in the number of VAChT/SP- and VIP-IR nerve fibers around the medullar arteries (A) was observed in cystic ovaries, while the number of VAChT-IR nerve endings near the cortical A was lowered after DXM application. Furthermore, nerve fibers containing VAChT were absent around veins in the whole ovary of DXM-treated animals. After DXM injections, an increase in the number of VAChT/SP- and VAChT/nNOS-IR nerve endings in the cortical, as well as VIP- and nNOS-IR (co-existing with VAChT), nerve terminals in the medullar part of the autonomic ground plexus (GP) was present. However, the administration of DXM led to a drop in the density of SOM-positive nerve endings (also VAChT-IR) in the medullar subdivision of the GP. The present study shows that in the porcine ovaries with DXM induced cysts the pattern of cholinergic innervation, as well as the co-localization of VAChT and NPY, VIP, SOM, SP or nNOS, were changed. Data obtained also suggest that acetylcholine and the above-mentioned neurotransmitters effecting the functioning (steroidogenic activity, blood flow) of the polycystic ovaries may have a significant influence on the course of this pathological status.
Multiple myeloma is a neoplastic disease which is characterised by proliferation of monoclonal plasmocytes in the bone marrow. It is the second most common hematologic cancer and it represents 1% of all cancer deaths. Despite enormous development in multiple myeloma biology and treatment over the last 30 years - it is still incurable disease with a median survival of 50 – 55 months. Currently, one of the most important goals in the treatment of multiple myeloma is to achieve long-term control of the disease, without negative impact on the patient’s quality of life. Thanks to therapeutic regimens based on new immunomodulatory drugs, this aim seems to be achievable. In this paper we present the case of a female patient living with multiple myeloma for 14 years. Initially patient was treated with standard VAD (vincristine, doxorubicin, dexamethasone) chemotherapy regimen. After a nearly complete remission of the disease, autotransplantation of hematopoietic cells was performed. One year after transplantation there was a relapse of the disease. In the treatment of relapse it was decided to use scheme based on lenalidomide and dexamethasone. After 4th cycle of treatment, a complete remission was achieved. So far, the patient received 149 cycles. In the evaluation of minimal residual disease still maintains a state of complete remission maintains. During over 12 years of treatment no complications in grade 3 and 4 of the CTCAE v.4 was observed. Currently the patient is 58 years old, she still receives lenalidomide and leads moderately active life.
The early molecular events of glucocorticoid-induced apoptosis have been investigated by studying glucocorticoid receptor levels, as well as binding activities to GRE and AP-1 sequences, using nuclear extracts from dexamethasone (Dex)-treated rat thymocytes. When the time-course of glucocorticoid-receptor complexes in nuclei of thymocytes was evaluated by binding studies using the tritiated ligand, we found that nuclear accumulation of radioactive complexes occurred in the first hour of incubation, and was followed by a progressive decline. This trend was confirmed by immunoblotting of nuclear proteins using a monoclonal anti-glucocorticoid receptor antibody. When the kinetics of binding activity to AP-1 and GRE sequences were studied, using nuclear extracts prepared from Dex-treated thymocytes in gel shift assays, we found peaks at 1 and 2 h after Dex treatment, and a return to basal levels in the following hours. Binding specificity was proved by competition studies using non-radioactive sequences, including mutated AP-1. Unexpectedly, however, protein binding to GRE was better competed for by AP-1 sequence than by GRE itself. Data obtained using the super gel shift assay suggested that AP-l/Jun can be responsible for the high affinity for the GRE sequence. Thus, we report here for the first time that an interference between AP-1 and GR in the binding to DNA consensus sequences — previously described in other biological systems — also occurs during apoptosis induced by glucocorticoids in lymphoid cells.
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