RESUMEN
AIM: Confirmation of taxonomic position of Lactobacillus fermentum 90 TC-4 strain using phenotypic (classic microbiological, MALDI TOF mass-spectrometry) and genetic (16S rRNA gene segment sequencing and full genome sequencing) methods. MATERIALS AND METHODS: Object of the study--Lactobacillus fermentum 90 TC-4 strains from various collections. Mass-spectrometric analysis was carried out using Autoflex MALDI TOF mass-spectrometer (Bruker Daltonics, Germany), study of biochemical properties of the strain was carried out using API 50 CHL strips (Biomerueux, France), "DNA-sorb B" kitwas used for isolation ofgenome DNA (CRIE, Moscow). Sequencing of the accumulated fragments of 16S rRNA gene was carried out using GenomeLab GeXP sequencing (Beckman Coulter, USA), full genome sequencing was carried out in MiSeq platform (Illumina). Assembly of genome and bioinformation analysis was carried out using BLAST program (www.blast.ncbi.nlm.nih.gov/blast.cgi), "CLC Bio Assembly" and genome server RAST (rast.nmpdr.org). RESULTS: L. fermentum 90 TC-4 strain was established to be contaminated by L. plantarum culture in a series of cases. As a result of identification of a pure culture of L. fermentum 90 TC-4 strain using a specter of high-technology methods, membership of the strain in L. fer- mentum species has been proven. CONCLUSION: Taxonomic status of L. fermentum 90 TC-4 strain was confirmed.
Asunto(s)
Limosilactobacillus fermentum/genética , Filogenia , Probióticos , ARN Ribosómico 16S/genética , Secuencia de Bases , Genoma Bacteriano , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Limosilactobacillus fermentum/clasificación , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
With the use of whole genome sequencing, the taxonomic status of Lactobacillus fermentum 90 TC-4 strain from Russian collections were studied. Complex analysis of phenotypical and genetic properties was conducted using phenotypic and molecular genetic methods. The main characteristics of the genome and biochemical activity profile of the strain were determined. A comparative analysis of the mass spectrum of ribosomal proteins of the strain, its biochemical properties, a fragment of 16S rRNA gene sequencing, and the entire genome revealed that the present strain belongs to the species L. fermentum, confirming its taxonomic status in accordance with modern taxonomy.
Asunto(s)
Limosilactobacillus fermentum , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Secuenciación Completa del Genoma , Limosilactobacillus fermentum/clasificación , Limosilactobacillus fermentum/genética , Limosilactobacillus fermentum/metabolismoRESUMEN
The analysis of efficiency of treatment of 17 patients with Gaucher disease (GD) in Ukraine who had received fermento-substitution therapy for 2 years and more was conducted on the basis of clinical and laboratory monitoring data. Regular infusions of recombinant glucocerebroside reduced signs of hepatosplenomegaly and pancytopenia, reduced a bone pain and a bone crisis at the majority of patients with GD I type that led to considerable improvement of health state and improvement of patients life quality. Efficiency of treatment depended on regularity of drug administration, dosage and severity level of the disease at the start of the therapy. Adult patients were not seen to have corrections of bones and neurologic disorders after the treatment that confirmed necessity of an early initiation of the treatment, before formation of irreversible changes in these organs and systems. Chitiotriodase activity in blood plasma is the most complex laboratory indicator which displays activity of pathological process in patients with GD, therefore it is necessary to use it for an estimation of treatment efficiency to correct a recombinant glucocerebroside dosage.
Asunto(s)
Biomarcadores/sangre , Terapia de Reemplazo Enzimático/métodos , Enfermedad de Gaucher , Glucosilceramidasa , Glucosilceramidas/sangre , Pancitopenia/sangre , Adolescente , Adulto , Huesos , Niño , Esquema de Medicación , Femenino , Enfermedad de Gaucher/sangre , Enfermedad de Gaucher/tratamiento farmacológico , Enfermedad de Gaucher/fisiopatología , Glucosilceramidasa/administración & dosificación , Glucosilceramidasa/deficiencia , Glucosilceramidasa/uso terapéutico , Hemoglobinas/análisis , Hepatomegalia/sangre , Hepatomegalia/fisiopatología , Humanos , Infusiones Intravenosas , Inyecciones Intravenosas , Masculino , Dolor , Calidad de Vida , Índice de Severidad de la Enfermedad , Esplenectomía , Esplenomegalia/sangre , Esplenomegalia/fisiopatología , Resultado del Tratamiento , UcraniaRESUMEN
AIM: The aim of the work is the development of laboratory test for indication and identification of Lactobacillus spp. by the polymerase chain reaction. MATERIALS AND METHODS: The work is developed on the base of the GenBank/EMBL data about genetic sequences of the Lactobacillus spp. The sequences of DNA were studied with the help of the ClustalW program. The strains of the Lactobacillus spp., which are the object of the research, have been registered in Russian collection of industrial microorganisms. RESULTS: The laboratory test of nested-PCR for indication and identification L. plantarum, L. fermentum, L. acidophilus, L. delbrueckii, L. casei, L. rhamnosus was performed. The specificity of the nested-PCR was correlated with the control analyses of monoculture Lactobacillus spp. and commercial products. CONCLUSION: The new developed laboratory nested-PCR test may be use in control system of milk foods enriched by probiotic microorganisms.
Asunto(s)
Lactobacillus/clasificación , Lactobacillus/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Probióticos/clasificación , Probióticos/aislamiento & purificación , Técnicas de Tipificación Bacteriana/métodos , Cartilla de ADN , Lactobacillus/genética , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Sensibilidad y Especificidad , Programas InformáticosRESUMEN
The paper considers how to obtain quantitative characteristics of sensomotor responses while tracing, learning, and reproducing rhythmical stimuli of different frequency and modality which can be used to assess psychomotor activity, perception, attention, short-term memory in schoolchildren with infantile cerebral paralysis. The results of examinations of children with this condition are given and analyzed.
Asunto(s)
Parálisis Cerebral/fisiopatología , Diagnóstico por Computador , Desempeño Psicomotor , Adolescente , Factores de Edad , Atención , Parálisis Cerebral/diagnóstico , Niño , Femenino , Humanos , Masculino , Memoria a Corto Plazo , Percepción , Factores de TiempoRESUMEN
3,5-Cyclic phosphates and phosphoramides of 6-halogenated glucofuranoses were synthesized via interaction of 3,5,6-bicyclophosphites of 1,2-O-alkylidene-alpha-D-glucofuranoses with halogens (followed by treatment with nucleophilic reagents) and N-chloroamines. 3,5-Cyclic trans-dibutylphosphoramides of 6-chloro-6-deoxy-1,2-O-isopropylidene- and 6-chloro-6-deoxy-(R)-(2,2,2)-trichloroethylidene)-alpha-D-glucofuranoses were shown to possess antiproliferative activity against CaOv human ovarian carcinoma cells in vitro (CE50 of approximately 10(-5) M). Cyclic trans-dibutylphosphoramide of 6-chloro-6-deoxy-1,2,-O-isopropylidene-alpha-D-glucofuranose also displayed marked antitumor effect on P-388 transplantable murine leukemia in vivo (the maximum increase in life span of 100% was reached at the quintuple injection of 100 mg/kg daily).
Asunto(s)
Antineoplásicos/síntesis química , Antineoplásicos/farmacología , Glucofosfatos/síntesis química , Leucemia P388/tratamiento farmacológico , Animales , Antineoplásicos/uso terapéutico , División Celular/efectos de los fármacos , Glucofosfatos/farmacología , Glucofosfatos/uso terapéutico , Leucemia P388/mortalidad , Espectroscopía de Resonancia Magnética , Ratones , Relación Estructura-Actividad , Células Tumorales CultivadasRESUMEN
A synthesis of 3-(1H-3-indolyl)-4-(1-glycosyl-3-indolyl)furan-2,5-diones and -1H-pyrrole-2,5-diones modified with the residues of D-ribo-, D-xylo-, L-arabino-, D-galactopyranose, and D-lactose was described. Influence of the compounds prepared on DNA biosynthesis in CaOv cells was studied.
Asunto(s)
Antineoplásicos/síntesis química , Glicósidos/síntesis química , Antineoplásicos/farmacología , División Celular/efectos de los fármacos , Glicósidos/farmacología , Humanos , Espectroscopía de Resonancia Magnética , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Interaction of 3'-amino-3'-deoxy-2',3'-secoadenosine with the N-hydroxysuccinimide esters of nicotinic or quinaldic acids and with 1-nitroanthraquinone-2-carboxylic acid in the presence of 2-ethoxy-1-ethoxy-carbonyl-1,2-dihydroquinoline led to the corresponding amides. To obtain 5'-modified 2',3'-secoadenosine analogs, 5'-deoxy-5'-nicotinoylamido-, 5'-deoxy-5'-(quinoline-2- carbonylamido)-, and 5'-deoxy-5'-[3-(3-indolyl)propionylamido]adenosine were subjected to the periodate oxidation--sodium borohydride reduction procedure. Structures of the synthesized compounds were was confirmed by 1H NMR spectroscopy, 2',3'-Diamino-2',3'-deoxy-, 3'-deoxy-3'-(quinoline-2-carbonylamido)-, and 5'-deoxy-5'-[3-(3-indolyl)propionylamido]-2',3'-secoadenosines+ ++ exhibited a cytotoxic effect on CaOv cells in vitro (CE50 10-30 microM).
Asunto(s)
Adenosina/análogos & derivados , Antineoplásicos/síntesis química , Acilación , Adenosina/síntesis química , Adenosina/farmacología , Antineoplásicos/farmacología , Femenino , Humanos , Espectroscopía de Resonancia Magnética , Neoplasias Ováricas/patología , Células Tumorales CultivadasRESUMEN
O-Quinaldoyl derivatives of thymidine, 2'-deoxyuridine, and 5-trimethylsilyl-2'-deoxyuridine were synthesized. 5'-Deoxy-5'-(quinoline-2-carbonylamino)- and 5'-deoxy-5'-[(quinoline-2-carbonylamino)butyroylamino]thymidine were obtained by the reaction of 5'-amino-5'-deoxythymidine with pentafluorophenyl ester of quinaldic acid, or with 4-(quinoline-2-carbonylamino)butyric acid. Antiproliferative properties in respect to CaOv cells in vitro were found in most of the synthesized quinaldoyl derivatives of nucleosides (CE50 approximately 10(-5) M). 3'-O-Quinaldoylthymidine exhibited an antitumor activity in vivo. The interaction of 3'- and 5'-O-quinaldoyl- as well as 3',5'-di-O-quinaldoylthymidine with DNA was investigated by the method of fluorescent probes.
Asunto(s)
Antineoplásicos/síntesis química , Desoxiuridina/análogos & derivados , Desoxiuridina/síntesis química , Quinolinas/química , Timidina/síntesis química , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , División Celular/efectos de los fármacos , Pollos , ADN/química , ADN/efectos de los fármacos , Desoxiuridina/química , Desoxiuridina/farmacología , Femenino , Humanos , Conformación de Ácido Nucleico , Neoplasias Ováricas/patología , Timidina/química , Timidina/farmacología , Células Tumorales CultivadasRESUMEN
The reaction of 5'-amino-2',5'-dideoxyuridine and 5'-amino-5'-deoxy-2',3'-O-ethoxymethyliden-6- azauridine with 3-(3-indolyl)propionic or 1-nitroanthraquinon-2-carboxylic acids in THF in the presence of 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) resulted in the corresponding amide derivatives. The reaction conditions of the standard procedure for the removal of the O-alkylidene protecting group turned out to be too severe for the 5'-N-acylamide derivatives of 6-azauridine. 5'-Deoxy-5'-[3-(3-indolyl)propionyl-amino]-6-azauridine was synthesized from 5'-amino-5'-deoxy-6-azauridine and 3-(3-indolyl)propionic acid in THF in the presence of EEDQ. A reaction between 5'-O-tosyl-2',3'-O-ethoxymethyliden-6-azauridine and 3-aminopropanol gave 3-(3-hydroxypropylamino)-2-(2',3'-O-ethoxymethylidene-beta- D-ribofuranosyl)-as-triazine-5(2H)-one, the structure of which was confirmed also by synthesis from O2,5'-anhydronucleoside and 3-aminopropanol followed by further chemical transformations. A reaction of 3-(3-hydroxypropylamino) derivative obtained with nicotinoyl chloride prepared in situ, or with 1-nirtoanthraquinon-2-carboxylic acid in the presence of DCC with subsequent deprotection, afforded 3-[(3-pyridin-3-ylcarboxy)propylamino]- or 3-[3-(1-nitroanthraquinon-2-carboxy)propylamino]-2-beta-D-ribof ura nosyl-as- triazine-5(2H)-one, respectively. Structures of the nucleosides prepared were examined by 1H NMR spectroscopy. 2',5'-Dideoxy-5'-[(1-nitroanthraquinon-2-carbonyl)amino]uridine at a 10(-4) M concentration was shown to inhibit thymidine incorporation into cell DNA (CE50 10(-5) M) by 72%.
Asunto(s)
Antraquinonas/química , Ácidos Carboxílicos/química , Desoxiuridina/química , Indoles/química , Ácidos Nicotínicos/química , Propionatos/química , Nucleósidos de Pirimidina/química , Aminas/química , Antineoplásicos/química , Antineoplásicos/farmacología , Desoxiuridina/farmacología , Humanos , Indicadores y Reactivos , Espectroscopía de Resonancia Magnética , Nucleósidos de Pirimidina/farmacología , Células Tumorales CultivadasRESUMEN
Interaction of nicotinoyl chloride in situ with 2'-deoxyuridine, its 3'-O-acetyl-, or 5'-O-trityl derivatives led to 3'-O-nicotinoyl-, 5'-O-nicotinoyl-, 3',5'-di-O-nicotinoyl-, and N3,3'-di-O-nicotinoyl-2'-deoxyuridine. Similarly, 5'-O-nicotinoyl-6-azauridine resulted from the reaction of 2',3'-O-ethoxymethylidene-6-azauridine followed by the deprotection. Reaction of 5'-amino-5'-deoxy-2',3'-O-ethoxymethylidene-6-azauridine with nicotinic acid in the presence of 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline followed by the cleavage of the 2',3'-O-protecting group gave 5'-deoxy-5'-nicotinamido-6-azauridine. The same compound was obtained from 5'-amino-5'-deoxy-6-azauridine and N-succinimidyl nicotinate. Structures of the compounds obtained were corroborated by 1H NMR spectra. It is shown that 3',5'-di-O-nicotinoyl-2'- deoxyuridine and 5'-deoxy-5'-O-nicotinamido-6-azauridine are cytotoxic toward CaOv cells in vitro (CE50 10(-5) M).
Asunto(s)
Antineoplásicos/química , Niacina/análogos & derivados , Nucleósidos de Pirimidina/química , Antineoplásicos/farmacología , Femenino , Humanos , Espectroscopía de Resonancia Magnética , Niacina/química , Neoplasias Ováricas/patología , Nucleósidos de Pirimidina/farmacología , Células Tumorales CultivadasRESUMEN
The effect of MESNA, an uroprotector (sodium 2-mercaptoethane sulfonate) on intracellular accumulation of doxorubicin and its interaction with DNA was studied in the cultures of the cells of human ovary carcinoma (CaOV) and normal epithelial kidney cells of green monkeys (VERO). The study was performed with methods developed by the authors. The results showed that MESNA had no effect on the qualitative and quantitative characteristics of the indicated processes in both the types of the doxorubicin sensitive cells. It was concluded that with the combined use of doxorubicin and phosphamide or cyclophosphane the use of MESNA for lowering the urotoxic action of oxazophosphorines had no effect on the biological efficacy of doxorubicin. The necessity of studying the possible modifying effect of MESNA in the cultures of doxorubicin resistant cells and in vivo is indicated.
Asunto(s)
ADN/metabolismo , Daunorrubicina/metabolismo , Mesna/farmacología , Animales , Chlorocebus aethiops , Humanos , Células Tumorales Cultivadas , Células VeroRESUMEN
Development, course and treatment of glomerulonephritis in comparison with other nephropathies was studied in 295 Hb-carriers with the nephrotic syndrome. Results of quantitative content of Hb-antigens in the blood serum and urine and nephrobiopsy materials were evaluated to show the etiopathogenetic role of Hb-viral infection in the development of glomerulonephritis. Four groups of Hb-carriers of the nephrotic syndrome were studied depending on hepatitis B markers in the blood serum. Treatment results are described in this category of patients.
Asunto(s)
Hepatitis B/complicaciones , Síndrome Nefrótico/etiología , Biomarcadores/química , Portador Sano/inmunología , Antígenos HLA/sangre , Hepatitis B/inmunología , Anticuerpos contra la Hepatitis B/análisis , Antígenos de la Hepatitis B/análisis , Humanos , Síndrome Nefrótico/inmunología , Renografía por RadioisótopoRESUMEN
The effect of different inorganic and organic selenium compounds on the nucleic acid synthesis in culture of human carcinoma ovary cells was studied. The rate of synthesis was determined by the incorporation of 3H-thymidine in the DNA and 3H-uridine in the RNA. Selenium compounds in the concentration to 1 mg/ml inhibited the nucleic acid synthesis in human tumour cells. The level of nucleic acid inhibition depends on the concentration of examined compounds. The tetravalent selenium compounds possessed the most pronounced cytotoxic effect.
Asunto(s)
Carcinoma/tratamiento farmacológico , Neoplasias Ováricas/tratamiento farmacológico , Selenio/toxicidad , Carcinoma/metabolismo , Células Cultivadas , ADN de Neoplasias/biosíntesis , Evaluación Preclínica de Medicamentos , Femenino , Humanos , Neoplasias Ováricas/metabolismo , ARN Neoplásico/biosíntesis , Selenio/uso terapéuticoRESUMEN
1-beta-D-Ribofuranosyl-4-methylthiopyrazolo(3,4-d)pyrimidine (I) has been converted into its 5'-monophosphate (III) by reacting with POCl3 in trialkyl phosphates or by phosphorylating 2',3'-O-ethoxymethylidene derivative of riboside (I) using 2-cyanoethyl phosphate in the presence of DCC and subsequent removal of blocking groups. Condensation of nucleotide (III) imidazolide with pyrophosphoric acid afforded corresponding 5'-triphosphate. Pools of natural NTPs and riboside (I) phosphates were monitored by HPLC after administering riboside (I), phosphate (III), or 4-methylthiopyrazolo(3,4-d)pyrimidine (II) into mice with leukemia L1210 or after incubating CaOv culture cells with these compounds. Treatment with riboside (I) or nucleotide (III) possessing antileukemic and cytotoxic activites led to much higher level of monophosphate (III), than treatment with biologically inactive base (II). ATP and GTP levels in CaOv cells incubated with (I) or (III) decreased by 60-70%, whereas (II) did not affect NTP pool. Bioactivation of nucleoside (I) into monophosphate (III) proceeds via direct phosphorylation by adenosine kinase. No tranformation of (II) into (I) or (III) occurs under these conditions.
Asunto(s)
Antineoplásicos/metabolismo , Leucemia L1210/metabolismo , Tionucleósidos/metabolismo , Tionucleótidos/metabolismo , Animales , Antineoplásicos/síntesis química , Biotransformación , Células Cultivadas , Fenómenos Químicos , Química , Femenino , Humanos , Ratones , Neoplasias Ováricas/metabolismo , Tionucleósidos/síntesis química , Tionucleótidos/síntesis químicaRESUMEN
The effect of glutamin (asparagin)ase from Pseudomonas fluorescens and Pseudomonas boreopolis 526 on DNA synthesis by tumor cells, lines CaOv and L8, has been studied. The L8 cells have been demonstrated highly sensitive to the enzyme.
Asunto(s)
Amidohidrolasas/farmacología , ADN de Neoplasias/biosíntesis , Pseudomonas/enzimología , Línea Celular , Relación Dosis-Respuesta a Droga , HumanosRESUMEN
The influence of methionine-gamma-lyase from Pseudomonas putida on DNA synthesis by CaOv and L-8 cell lines has been studied. The agent has been demonstrated to inhibit the incorporation of 3H-thymidine into L-8 cell line and to have no effect on CaOv cells.