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1.
Clin Microbiol Infect ; 26(3): 384.e5-384.e8, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31705996

ABSTRACT

OBJECTIVE: Laboratory quality control (QC) is essential to assess the reliability of tuberculosis diagnostic testing. To provide safe QC reagents for the detection of drug-resistant Mycobacterium tuberculosis, we generated antibiotic-resistant mycobacterial strains of attenuated virulence (M. bovis bacillus Calmette-Guérin (BCG)). METHODS: Seven mono-resistant BCG strains were developed by introducing resistance-conferring mutations into wild-type BCG strains. Mutations were confirmed by dideoxynucleotide sequencing. Phenotypic resistance was quantified by microbroth dilution to determine the MIC90. The capacity of two commercial tests (GeneXpert TB/RIF and Genotype MTBDRplus) to detect resistance-conferring mutations was evaluated independently. RESULTS: Our panel included BCG strains with mutations in rpoB (S450L, I491F), katG (deletion at AA428), gyrA (D94G), rpsL (K43R) and Rv0678c (S63R). These mutations translated respectively into phenotypic resistance to rifampin (MIC ≥8 mg/L), isoniazid (MIC ≥8 mg/L), moxifloxacin (MIC 4 mg/L) and streptomycin (MIC ≥8 mg/L); the Rv0678c mutant showed decreased susceptibility to both clofazimine (MIC 4 mg/L) and bedaqualine (MIC 1 mg/L). GeneXpert (Cepheid) and Genotype MTBDRplus (Hain Lifesciences) both called the rpoB S450L strain rifampin-resistant and the I491F mutant rifampin-susceptible, as expected based on single nucleotide polymorphism positions. Likewise, MTBDRplus called the novel katG deletion mutant isoniazid susceptible despite phenotypic resistance. CONCLUSION: BCG strains engineered to be mono-resistant to anti-tuberculosis drugs can be used as safe QC reagents for tuberculosis diagnostics and drug susceptibility testing.


Subject(s)
Antitubercular Agents/pharmacology , Drug Resistance, Multiple, Bacterial , Mutation , Mycobacterium bovis/drug effects , Mycobacterium bovis/genetics , Tuberculosis, Bovine/diagnosis , Tuberculosis, Bovine/microbiology , Alleles , Amino Acid Substitution , Animals , Bacterial Proteins/genetics , Cattle , Clinical Laboratory Techniques/methods , Clinical Laboratory Techniques/standards , Codon , Dose-Response Relationship, Drug , Genotype , Humans , Mycobacterium bovis/classification , Polymorphism, Single Nucleotide , Quality Control , Rifampin/pharmacology , Tuberculosis, Bovine/drug therapy
2.
Placenta ; 29(3): 266-73, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18279954

ABSTRACT

Marinobufagenin (MBG) is an endogenous mammalian cardiotonic steroid that is involved in the inhibition of the sodium pump Na(+)/K(+)-ATPase. Increased plasma levels of MBG have been reported in patients with volume expansion-mediated hypertension and preeclampsia. We have recently demonstrated that MBG impairs both the proliferation and growth factor-induced migration of human first trimester cytotrophoblast (CTB) cells, crucial for proper placental development. However, the intracellular signaling mechanisms regulating the MBG-induced impairment of CTB differentiation, migration and invasion are unknown. The human extravillous CTB cell line SGHPL-4 was utilized for this study. The phosphorylation of MAP kinase protein ERK1/2 was evaluated by Cellular Activation of Signaling ELISA (CASE) in control CTB cells and those treated with MBG. MBG at concentrations of 10 and 100nM inhibited CTB cell proliferation, migration and invasion (60%, 50% and 50%, respectively). MBG also caused a significant decrease in the phosphorylation of ERK1/2. In addition, MBG decreased proliferation, migration, and ERK1/2 activity in another motile cell line, CHO cells. Another sodium pump inhibitor, ouabain, similarly decreased proliferation and ERK1/2 activity in CTB and CHO cells. These data suggest that the changes observed in cell function may be mediated by inhibition of Na(+)/K(+)-ATPase. We demonstrate that the MBG-induced impairment of CTB cell proliferation, migration and invasion is associated with decreased ERK1/2 activity which may be mediated by inhibition of Na(+)/K(+)-ATPase.


Subject(s)
Bufanolides/pharmacology , Cell Movement/drug effects , Cell Proliferation/drug effects , Trophoblasts/drug effects , Animals , CHO Cells , Cell Adhesion/drug effects , Cricetinae , Cricetulus , Enzyme Inhibitors/pharmacology , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Ouabain/pharmacology , Phosphorylation/drug effects , Sodium-Potassium-Exchanging ATPase/antagonists & inhibitors , Trophoblasts/physiology
3.
Am J Nephrol ; 28(1): 8-13, 2008.
Article in English | MEDLINE | ID: mdl-17890853

ABSTRACT

BACKGROUND/AIMS: There are two major pathophysiologic processes involved in the development of hypertension: (1) expanded extracellular fluid volume and (2) vasoconstriction. We have developed a model of preeclampsia in the rat, in which excessive volume expansion (VE) plays a role. These animals excrete increased amounts of the bufodienolide, marinobufagenin (MBG), even before their hypertension and proteinuria become established. Furthermore, their hypertension is corrected by administration of resibufogenin (RBG), a compound structurally similar to MBG. METHOD: We studied two models of experimental hypertension in the nonpregnant animal, produced either by deoxycorticosterone acetate (DOCA)-salt administration or by angiotensin infusion. RESULTS: RBG administered to the DOCA-salt rats lowered blood pressure and reduced proteinuria in the VE animals, but had no affect on the rats infused with angiotensin. Furthermore, although the production of superoxide anion in the aortas of both groups of hypertensive rats was increased over control, RBG reduced these levels to normal in the VE (DOCA-salt) animals only. RBG had no effect in the angiotensin-infused rats. The urinary excretion of angiotensinogen did not rise in VE-mediated hypertension, but did increase in the angiotensin-infused rats. CONCLUSIONS: MBG plays an important role in the causation of hypertension in the VE rats, but not in the vasoconstrictive model. RBG is effective only in VE-mediated hypertension.


Subject(s)
Blood Pressure/drug effects , Bufanolides/pharmacology , Hypertension, Renal/drug therapy , Proteinuria/drug therapy , Angiotensin II/pharmacology , Angiotensinogen/urine , Animals , Bufanolides/antagonists & inhibitors , Creatinine/blood , Desoxycorticosterone , Disease Models, Animal , Hypertension, Renal/chemically induced , Male , Mineralocorticoids , Proteinuria/chemically induced , Rats , Rats, Inbred Strains , Sodium Chloride , Superoxides/metabolism , Vasoconstrictor Agents/pharmacology
4.
Klin Khir ; (11-12): 95-8, 2007.
Article in Ukrainian | MEDLINE | ID: mdl-18402039

ABSTRACT

The main and immediate causes of death, pathogenetic mechanisms of postoperative cerebral inflammatory-purulent complications (CIPC) in patients, suffering the brain tumour, were analyzed. The main pathogenetic patterns of the postoperative period course in the CIPC occurrence were studied, basing on the data of analysis of 30 patients, who died after the operation. There was proved, that the sepsis occurrence on the background of postoperative meningoencephalitis means the lethal ending of the disease. There was established, that in occurrence of septic complications, connected with postoperative meningoencephalitis, the most frequently the affection of respiratory and urinary systems occurs.


Subject(s)
Brain Neoplasms/surgery , Meningoencephalitis/etiology , Neurosurgical Procedures/adverse effects , Sepsis/etiology , Adolescent , Adult , Aged , Brain Neoplasms/mortality , Brain Neoplasms/pathology , Female , Humans , Male , Meningoencephalitis/mortality , Meningoencephalitis/pathology , Middle Aged , Neurosurgical Procedures/mortality , Sepsis/mortality , Sepsis/pathology
5.
Biofizika ; 39(2): 323-7, 1994.
Article in Russian | MEDLINE | ID: mdl-8193193

ABSTRACT

The physical parameters of liposomes from phosphatidylcholine with cholesterol or dicetylphosphate were investigated. The sizes, electrophoretic mobility, surface density charges of these liposomes were estimated. It has been shown that dicetylphosphate, as well as cholesterol increases the thickness of bilayer of the lecithin liposomes and at the same time the area occupied by one lipid molecule has not been changed by dicetylphosphate in contrast to cholesterol.


Subject(s)
Cholesterol/pharmacology , Liposomes/chemistry , Organophosphates/pharmacology , Lipid Bilayers , Phosphatidylcholines/chemistry
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