11 research outputs found
ΠΠΏΡΠΈΠΌΠΈΠ·Π°ΡΠΈΡ ΡΠ΅Ρ Π½ΠΎΠ»ΠΎΠ³ΠΈΠΈ ΠΏΠΎΠ»ΡΡΠ΅Π½ΠΈΡ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ° Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° Π΄Π»Ρ Π±ΠΈΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠΉ ΠΊΠΎΡΡΠ΅ΠΊΡΠΈΠΈ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΡ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ°
Fecal microbiota transplantation (FMT) is now considered as an effective tool for the treatment of various GI pathologies. Fecal preparations are delivered both through the lower GIT (enema, colonoscopy) and upper (endoscopy, capsules). A common disadvantage of instrumental methods of administration is their high invasiveness associated with the risk of intestinal perforation and the use of anesthesia. Oral capsules are minimally invasive, comfortable and more aesthetic, so this method of drug delivery is gaining popularity. The main issue with the use of frozen feces (including the lyophilisate used in capsules) is its efficiency compared to the original material. During lyophilization, cells are exposed to stress factors such as low temperatures, water crystallization, osmotic stress, changes in pH, and dehydration. To reduce the likelihood of cell damage during lyophilization, protective media (lyo-protectants) are used. In this work sucrose, gelatin, and their combinations have been used as lyoprotectors. To estimate the number of microorganisms, a bacteriological study was carried out. The number of Bifidobacteria, Lactobacilli, and the total number of E.coli and Enterobacteriaceae was estimated. It was found that the lyophilized stool sample containing 10% sucrose as a protective medium had the highest number of viable cells. Also, the physical properties of the lyophilisate (its flowability) are convenient for preparing capsulated form. The molar ratios of short chain fatty acids (SCFAs) in the original fecal samples and lyophilisates were studied by gas chromatography. The molar ratios of major SCFAs (acetate, propionate and butyrate) were identical in the samples studied. The composition of the protective medium in which the lyophilized biomaterial corresponds to the original feces in terms of the number of "live" microorganisms has been proposed. According to its physical characteristics lyophilisate is convenient for capsules preparation.Π Π½Π°ΡΡΠΎΡΡΠ΅ΠΌΡ ΠΌΠΎΠΌΠ΅Π½ΡΡ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΡΡΠ°Π½ΡΠΏΠ»Π°Π½ΡΠ°ΡΠΈΠΈ ΡΠ΅ΠΊΠ°Π»ΡΠ½ΠΎΠΉ ΠΌΠΈΠΊΡΠΎΠ±ΠΈΠΎΡΡ (Π’Π€Π) ΠΏΡΠΈ Π»Π΅ΡΠ΅Π½ΠΈΠΈ ΡΠ°Π·Π»ΠΈΡΠ½ΡΡ
ΠΏΠ°ΡΠΎΠ»ΠΎΠ³ΠΈΠΉ ΠΠΠ’ Π½Π΅ Π²ΡΠ·ΡΠ²Π°Π΅Ρ ΡΠΎΠΌΠ½Π΅Π½ΠΈΠΉ. ΠΡΠ΅ΠΏΠ°ΡΠ°ΡΡ ΡΠ΅ΠΊΠ°Π»ΠΈΠΉ Π΄ΠΎΡΡΠ°Π²Π»ΡΡΡ ΠΊΠ°ΠΊ ΡΠ΅ΡΠ΅Π· Π½ΠΈΠΆΠ½ΠΈΠ΅ ΠΎΡΠ΄Π΅Π»Ρ ΠΠΠ’ (ΠΊΠ»ΠΈΠ·ΠΌΠ°, ΠΊΠΎΠ»ΠΎΠ½ΠΎΡΠΊΠΎΠΏΠΈΡ), ΡΠ°ΠΊ ΠΈ Π²Π΅ΡΡ
Π½ΠΈΠ΅ (ΡΠ½Π΄ΠΎΡΠΊΠΎΠΏΠΈΡ, ΠΊΠ°ΠΏΡΡΠ»Ρ). ΠΠ±ΡΠΈΠΌ Π½Π΅Π΄ΠΎΡΡΠ°ΡΠΊΠΎΠΌ ΠΈΠ½ΡΡΡΡΠΌΠ΅Π½ΡΠ°Π»ΡΠ½ΡΡ
ΠΌΠ΅ΡΠΎΠ΄ΠΎΠ² Π²Π²Π΅Π΄Π΅Π½ΠΈΡ ΡΠ²Π»ΡΠ΅ΡΡΡ ΠΈΡ
Π²ΡΡΠΎΠΊΠ°Ρ ΠΈΠ½Π²Π°Π·ΠΈΠ²Π½ΠΎΡΡΡ, ΡΠ²ΡΠ·Π°Π½Π½Π°Ρ Ρ ΡΠΈΡΠΊΠΎΠΌ ΠΏΠ΅ΡΡΠΎΡΠ°ΡΠΈΠΈ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ° ΠΈ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ΠΌ Π°Π½Π΅ΡΡΠ΅Π·ΠΈΠΈ. ΠΠ΅ΡΠΎΡΠ°Π»ΡΠ½ΡΠ΅ ΠΊΠ°ΠΏΡΡΠ»Ρ ΠΌΠΈΠ½ΠΈΠΌΠ°Π»ΡΠ½ΠΎ ΠΈΠ½Π²Π°Π·ΠΈΠ²Π½Ρ, ΡΠ΄ΠΎΠ±Π½Ρ ΠΈ Π±ΠΎΠ»Π΅Π΅ ΡΡΡΠ΅ΡΠΈΡΠ½Ρ, ΠΏΠΎΡΡΠΎΠΌΡ ΡΡΠΎΡ ΡΠΏΠΎΡΠΎΠ± Π΄ΠΎΡΡΠ°Π²ΠΊΠΈ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ° ΡΡΠ°Π½ΠΎΠ²ΠΈΡΡΡ Π²ΡΠ΅ Π±ΠΎΠ»Π΅Π΅ ΠΏΠΎΠΏΡΠ»ΡΡΠ½ΡΠΌ. ΠΡΠ½ΠΎΠ²Π½ΠΎΠΉ Π²ΠΎΠΏΡΠΎΡ, ΡΠ²ΡΠ·Π°Π½Π½ΡΠΉ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ Π·Π°ΠΌΠΎΡΠΎΠΆΠ΅Π½Π½ΠΎΠ³ΠΎ ΠΊΠ°Π»Π° (Π² ΡΠΎΠΌ ΡΠΈΡΠ»Π΅ Π»ΠΈΠΎΡΠΈΠ»ΠΈΠ·Π°ΡΠ°, ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΠ΅ΠΌΠΎΠ³ΠΎ Π² ΠΊΠ°ΠΏΡΡΠ»Π°Ρ
), Π·Π°ΠΊΠ»ΡΡΠ°Π΅ΡΡΡ Π² ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΡΠ°ΠΊΠΎΠ³ΠΎ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ° ΠΏΠΎ ΡΡΠ°Π²Π½Π΅Π½ΠΈΡ Ρ ΠΈΡΡ
ΠΎΠ΄Π½ΡΠΌ ΠΌΠ°ΡΠ΅ΡΠΈΠ°Π»ΠΎΠΌ. Π ΠΏΡΠΎΡΠ΅ΡΡΠ΅ Π»ΠΈΠΎΡΠΈΠ»ΠΈΠ·Π°ΡΠΈΠΈ ΠΊΠ»Π΅ΡΠΊΠΈ ΠΏΠΎΠ΄Π²Π΅ΡΠ³Π°ΡΡΡΡ Π΄Π΅ΠΉΡΡΠ²ΠΈΡ ΡΡΡΠ΅ΡΡΠΎΠ²ΡΡ
ΡΠ°ΠΊΡΠΎΡΠΎΠ², ΡΠ°ΠΊΠΈΡ
ΠΊΠ°ΠΊ Π½ΠΈΠ·ΠΊΠΈΠ΅ ΡΠ΅ΠΌΠΏΠ΅ΡΠ°ΡΡΡΡ, ΠΊΡΠΈΡΡΠ°Π»Π»ΠΈΠ·Π°ΡΠΈΡ Π²ΠΎΠ΄Ρ, ΠΎΡΠΌΠΎΡΠΈΡΠ΅ΡΠΊΠΈΠΉ ΡΡΡΠ΅ΡΡ, ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ ΡΠ ΡΠ°ΡΡΠ²ΠΎΡΠΎΠ², Π΄Π΅Π³ΠΈΠ΄ΡΠ°ΡΠ°ΡΠΈΡ. ΠΠ»Ρ ΡΠ½ΠΈΠΆΠ΅Π½ΠΈΡ ΡΠΈΡΠΊΠ° ΠΏΠΎΠ²ΡΠ΅ΠΆΠ΄Π΅Π½ΠΈΠΉ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΏΡΠΈ Π»ΠΈΠΎΡΠΈΠ»ΠΈΠ·Π°ΡΠΈΠΈ ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΡΡ Π·Π°ΡΠΈΡΠ½ΡΠ΅ ΡΡΠ΅Π΄Ρ (Π»ΠΈΠΎΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡΡ). Π ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅ Π»ΠΈΠΎΠΏΡΠΎΡΠ΅ΠΊΡΠΎΡΠΎΠ² Π² Π΄Π°Π½Π½ΠΎΠΉ ΡΠ°Π±ΠΎΡΠ΅ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π»ΠΈ ΡΠ°Ρ
Π°ΡΠΎΠ·Ρ, ΠΆΠ΅Π»Π°ΡΠΈΠ½ ΠΈ ΠΈΡ
ΠΊΠΎΠΌΠ±ΠΈΠ½Π°ΡΠΈΠΈ. ΠΠ»Ρ ΠΎΡΠ΅Π½ΠΊΠΈ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π° ΠΌΠΈΠΊΡΠΎΠΎΡΠ³Π°Π½ΠΈΠ·ΠΌΠΎΠ² ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ Π±Π°ΠΊΡΠ΅ΡΠΈΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ΅ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅. ΠΡΠ΅Π½ΠΈΠ²Π°Π»ΠΈ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²ΠΎ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ ΡΠΎΠ΄Π° Bifidobacterium, Lactobacillus, Escherichia, Π° ΡΠ°ΠΊΠΆΠ΅ ΡΠ΅ΠΌΠ΅ΠΉΡΡΠ²Π° Enterobacterales Π² ΡΠ΅Π»ΠΎΠΌ. Π£ΡΡΠ°Π½ΠΎΠ²Π»Π΅Π½ΠΎ, ΡΡΠΎ Π² Π»ΠΈΠΎΡΠΈΠ»ΠΈΠ·ΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΌ ΠΎΠ±ΡΠ°Π·ΡΠ΅ ΠΊΠ°Π»Π°, ΡΠΎΠ΄Π΅ΡΠΆΠ°ΡΠ΅ΠΌ Π² ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅ Π·Π°ΡΠΈΡΠ½ΠΎΠΉ ΡΡΠ΅Π΄Ρ 10 % ΡΠ°Ρ
Π°ΡΠΎΠ·Ρ, Π½Π°Π±Π»ΡΠ΄Π°Π΅ΡΡΡ Π½Π°ΠΈΠ±ΠΎΠ»ΡΡΠ΅Π΅ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²ΠΎ ΠΆΠΈΠ·Π½Π΅ΡΠΏΠΎΡΠΎΠ±Π½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ, ΡΠΈΠ·ΠΈΡΠ΅ΡΠΊΠΈΠ΅ ΡΠ²ΠΎΠΉΡΡΠ²Π° Π»ΠΈΠΎΡΠΈΠ»ΠΈΠ·Π°ΡΠ° (Π΅Π³ΠΎ ΡΡΠΏΡΡΠ΅ΡΡΡ) ΡΠ΄ΠΎΠ±Π½Ρ Π΄Π»Ρ Π½Π°ΠΏΠΎΠ»Π½Π΅Π½ΠΈΡ ΠΊΠ°ΠΏΡΡΠ». ΠΠ΅ΡΠΎΠ΄ΠΎΠΌ Π³Π°Π·ΠΎΠ²ΠΎΠΉ Ρ
ΡΠΎΠΌΠ°ΡΠΎΠ³ΡΠ°ΡΠΈΠΈ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½Ρ ΠΌΠΎΠ»ΡΡΠ½ΡΠ΅ ΡΠΎΠΎΡΠ½ΠΎΡΠ΅Π½ΠΈΡ ΠΠΠ Π² ΠΈΡΡ
ΠΎΠ΄Π½ΡΡ
ΠΎΠ±ΡΠ°Π·ΡΠ°Ρ
ΠΊΠ°Π»Π° ΠΈ Π»ΠΈΠΎΡΠΈΠ»ΠΈΠ·Π°ΡΠ°Ρ
. ΠΠΎΠ»ΡΡΠ½ΡΠ΅ ΡΠΎΠΎΡΠ½ΠΎΡΠ΅Π½ΠΈΡ ΠΌΠ°ΠΆΠΎΡΠ½ΡΡ
ΠΊΠΎΡΠΎΡΠΊΠΎΡΠ΅ΠΏΠΎΡΠ΅ΡΠ½ΡΡ
ΠΆΠΈΡΠ½ΡΡ
ΠΊΠΈΡΠ»ΠΎΡ (ΠΠΠ) Π°ΡΠ΅ΡΠ°ΡΠ°, ΠΏΡΠΎΠΏΠΈΠΎΠ½Π°ΡΠ° ΠΈ Π±ΡΡΠΈΡΠ°ΡΠ° ΠΎΠΊΠ°Π·Π°Π»ΠΈΡΡ ΠΈΠ΄Π΅Π½ΡΠΈΡΠ½Ρ Π² ΠΈΡΡΠ»Π΅Π΄ΡΠ΅ΠΌΡΡ
ΠΎΠ±ΡΠ°Π·ΡΠ°Ρ
. ΠΡΠ΅Π΄Π»ΠΎΠΆΠ΅Π½ ΡΠΎΡΡΠ°Π² Π·Π°ΡΠΈΡΠ½ΠΎΠΉ ΡΡΠ΅Π΄Ρ, Π² ΠΊΠΎΡΠΎΡΠΎΠΉ Π»ΠΈΠΎΡΠΈΠ»ΠΈΠ·ΠΈΡΠΎΠ²Π°Π½Π½ΡΠΉ Π±ΠΈΠΎΠΌΠ°ΡΠ΅ΡΠΈΠ°Π» ΠΌΠ°ΠΊΡΠΈΠΌΠ°Π»ΡΠ½ΠΎ ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΡΠ΅Ρ ΠΈΡΡ
ΠΎΠ΄Π½ΠΎΠΌΡ ΠΊΠ°Π»Ρ ΠΏΠΎ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Ρ Β«ΠΆΠΈΠ²ΡΡ
Β» ΠΌΠΈΠΊΡΠΎΠΎΡΠ³Π°Π½ΠΈΠ·ΠΌΠΎΠ². ΠΠΈΠΎΡΠΈΠ»ΠΈΠ·Π°Ρ ΠΏΠΎ ΡΠ²ΠΎΠΈΠΌ ΡΠΈΠ·ΠΈΡΠ΅ΡΠΊΠΈΠΌ Ρ
Π°ΡΠ°ΠΊΡΠ΅ΡΠΈΡΡΠΈΠΊΠ°ΠΌ ΡΠ΄ΠΎΠ±Π΅Π½ Π΄Π»Ρ ΠΏΡΠΈΠ³ΠΎΡΠΎΠ²Π»Π΅Π½ΠΈΡ ΠΊΠ°ΠΏΡΡΠ»
ΠΠΈΠΊΡΠΎΠ±ΠΈΠΎΠΌ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ° ΠΈ ΠΌΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΠ·ΠΌ Π»Π΅ΠΊΠ°ΡΡΡΠ²Π΅Π½Π½ΡΡ ΡΠΎΠ΅Π΄ΠΈΠ½Π΅Π½ΠΈΠΉ
The human physiology textbooks traditionally consider the intestine as a metabolically active organ, with its activity primarily associated with the production of numerous digestive enzymes. The development of molecular analysis technologies has significantly detailized this picture, primarily by decoding the metabolic potential of the intestinal microbiota. Data from numerous metagenomic studies indicate that the number of eukaryotic and bacterial cells in the human body is comparable - about 3.0Γ1013, while the number of genes in the intestinal metagenome is one hundred times greater than in the human genome. Obviously, the gut microbiota exhibits both direct and indirect effects on the metabolism of drugs and xenobiotics, that can affect their effectiveness and toxicity. Orally administrated xenobiotics have been found to be metabolized by intestinal microbial enzymes before being absorbed from the gastrointestinal tract into the blood flow. The metabolic reactions performed by the gut microbiota greatly differ from the metabolic reactions of the liver, providing modification of drugs by acetylation, deacetylation, decarboxylation, dehydroxylation, demethylation, dehalogenation, etc. Despite the metabolism of xenobiotics by microbial enzymes of the intestine is rather known, information about the specific microflora mediating each metabolic reaction is still limited, mainly by the lack of an adequate model of the intestinal microbial community to allow the accumulation of experimental data for the creation of computational models. Currently, studies of drug metabolism use microfluidic chips, reproducing functions of various organs and tissues, such as the liver, kidney, lungs and intestine, as in vitro models in the form of 2D and 3D cell cultures. Supplementation of such systems with the microbial community will allow to get as close as possible to in vitro modeling of complicated biological processes in the interests of pharmacological research and the accumulation of data for constructing computational models.Π ΠΊΡΡΡΠ΅ ΡΠΈΠ·ΠΈΠΎΠ»ΠΎΠ³ΠΈΠΈ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊ ΡΡΠ°Π΄ΠΈΡΠΈΠΎΠ½Π½ΠΎ ΡΠ°ΡΡΠΌΠ°ΡΡΠΈΠ²Π°ΡΡ ΠΊΠ°ΠΊ ΠΌΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΡΠ΅ΡΠΊΠΈ Π°ΠΊΡΠΈΠ²Π½ΡΠΉ ΠΎΡΠ³Π°Π½, Π΄Π΅ΡΡΠ΅Π»ΡΠ½ΠΎΡΡΡ ΠΊΠΎΡΠΎΡΠΎΠ³ΠΎ ΡΠ²ΡΠ·ΡΠ²Π°ΡΡ Π² ΠΏΠ΅ΡΠ²ΡΡ ΠΎΡΠ΅ΡΠ΅Π΄Ρ Ρ ΠΏΡΠΎΠ΄ΡΠΊΡΠΈΠ΅ΠΉ ΠΌΠ½ΠΎΠ³ΠΎΡΠΈΡΠ»Π΅Π½Π½ΡΡ
ΠΏΠΈΡΠ΅Π²Π°ΡΠΈΡΠ΅Π»ΡΠ½ΡΡ
ΡΠ΅ΡΠΌΠ΅Π½ΡΠΎΠ². Π Π°Π·Π²ΠΈΡΠΈΠ΅ ΡΠ΅Ρ
Π½ΠΎΠ»ΠΎΠ³ΠΈΠΉ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΠΎΠ³ΠΎ Π°Π½Π°Π»ΠΈΠ·Π° ΠΏΠΎΠ·Π²ΠΎΠ»ΠΈΠ»ΠΎ ΡΡΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎ Π΄Π΅ΡΠ°Π»ΠΈΠ·ΠΈΡΠΎΠ²Π°ΡΡ ΡΡΡ ΠΊΠ°ΡΡΠΈΠ½Ρ, Π² ΠΏΠ΅ΡΠ²ΡΡ ΠΎΡΠ΅ΡΠ΅Π΄Ρ Π·Π° ΡΡΠ΅Ρ ΡΠ°ΡΡΠΈΡΡΠΎΠ²ΠΊΠΈ ΠΌΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΠΏΠΎΡΠ΅Π½ΡΠΈΠ°Π»Π° ΠΊΠΈΡΠ΅ΡΠ½ΠΎΠΉ ΠΌΠΈΠΊΡΠΎΠ±ΠΈΠΎΡΡ. ΠΠ°Π½Π½ΡΠ΅ ΠΌΠ½ΠΎΠ³ΠΎΡΠΈΡΠ»Π΅Π½Π½ΡΡ
ΠΌΠ΅ΡΠ°Π³Π΅Π½ΠΎΠΌΠ½ΡΡ
ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠΉ ΡΠ²ΠΈΠ΄Π΅ΡΠ΅Π»ΡΡΡΠ²ΡΡΡ, ΡΡΠΎ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²ΠΎ ΡΡΠΊΠ°ΡΠΈΠΎΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΈ Π±Π°ΠΊΡΠ΅ΡΠΈΠ°Π»ΡΠ½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π² ΠΎΡΠ³Π°Π½ΠΈΠ·ΠΌΠ΅ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° ΡΠΎΠΏΠΎΡΡΠ°Π²ΠΈΠΌΠΎ β ΠΎΠΊΠΎΠ»ΠΎ 3.0Ρ
1013, ΠΏΡΠΈ ΡΡΠΎΠΌ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²ΠΎ Π³Π΅Π½ΠΎΠ² Π² ΠΌΠ΅ΡΠ°Π³Π΅Π½ΠΎΠΌΠ΅ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ° Π² ΡΡΠΎ ΡΠ°Π· Π±ΠΎΠ»ΡΡΠ΅, ΡΠ΅ΠΌ Π² Π³Π΅Π½ΠΎΠΌΠ΅ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ°. ΠΡΠ΅Π²ΠΈΠ΄Π½ΠΎ, ΡΡΠΎ ΠΌΠΈΠΊΡΠΎΠ±ΠΈΠΎΡΠ° ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ° ΠΎΠΊΠ°Π·ΡΠ²Π°Π΅Ρ ΠΊΠ°ΠΊ ΠΏΡΡΠΌΠΎΠ΅, ΡΠ°ΠΊ ΠΈ ΠΎΠΏΠΎΡΡΠ΅Π΄ΠΎΠ²Π°Π½Π½ΠΎΠ΅ Π²Π»ΠΈΡΠ½ΠΈΠ΅ Π½Π° ΠΌΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΠ·ΠΌ Π»Π΅ΠΊΠ°ΡΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ² ΠΈ ΠΊΡΠ΅Π½ΠΎΠ±ΠΈΠΎΡΠΈΠΊΠΎΠ², ΡΡΠΎ ΠΌΠΎΠΆΠ΅Ρ ΡΠΊΠ°Π·Π°ΡΡΡΡ Π½Π° ΠΈΡ
ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΠΈ ΡΠΎΠΊΡΠΈΡΠ½ΠΎΡΡΠΈ. ΠΠ±Π½Π°ΡΡΠΆΠ΅Π½ΠΎ, ΡΡΠΎ ΠΊΡΠ΅Π½ΠΎΠ±ΠΈΠΎΡΠΈΠΊΠΈ, Π²Π²ΠΎΠ΄ΠΈΠΌΡΠ΅ ΠΏΠ΅ΡΠΎΡΠ°Π»ΡΠ½ΠΎ, ΠΌΠΎΠ³ΡΡ ΠΌΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΠ·ΠΈΡΠΎΠ²Π°ΡΡΡΡ ΠΊΠΈΡΠ΅ΡΠ½ΡΠΌΠΈ ΠΌΠΈΠΊΡΠΎΠ±Π½ΡΠΌΠΈ ΡΠ΅ΡΠΌΠ΅Π½ΡΠ°ΠΌΠΈ Π΅ΡΠ΅ Π΄ΠΎ Π²ΡΠ°ΡΡΠ²Π°Π½ΠΈΡ ΠΈΠ· ΠΆΠ΅Π»ΡΠ΄ΠΎΡΠ½ΠΎ-ΠΊΠΈΡΠ΅ΡΠ½ΠΎΠ³ΠΎ ΡΡΠ°ΠΊΡΠ° Π² ΠΊΡΠΎΠ²Ρ. ΠΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΡΠ΅ΡΠΊΠΈΠ΅ ΡΠ΅Π°ΠΊΡΠΈΠΈ, Π²ΡΠΏΠΎΠ»Π½ΡΠ΅ΠΌΡΠ΅ ΠΌΠΈΠΊΡΠΎΠ±ΠΈΠΎΡΠΎΠΉ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ°, Π·Π½Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎ ΠΎΡΠ»ΠΈΡΠ°ΡΡΡΡ ΠΎΡ ΠΌΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΡΠ΅ΡΠΊΠΈΡ
ΡΠ΅Π°ΠΊΡΠΈΠΉ ΠΏΠ΅ΡΠ΅Π½ΠΈ, ΠΎΠ±Π΅ΡΠΏΠ΅ΡΠΈΠ²Π°Ρ ΠΌΠΎΠ΄ΠΈΡΠΈΠΊΠ°ΡΠΈΡ Π»Π΅ΠΊΠ°ΡΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ² ΠΏΡΡΠ΅ΠΌ Π°ΡΠ΅ΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ, Π΄Π΅Π°ΡΠ΅ΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ, Π΄Π΅ΠΊΠ°ΡΠ±ΠΎΠΊΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ, Π΄Π΅Π³ΠΈΠ΄ΡΠΎΠΊΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ, Π΄Π΅ΠΌΠ΅ΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ, Π΄Π΅Π³Π°Π»ΠΎΠ³Π΅Π½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΈ Π΄Ρ. ΠΠ΅ΡΠΌΠΎΡΡΡ Π½Π° ΡΠΎ, ΡΡΠΎ ΠΌΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΠ·ΠΌ ΠΊΡΠ΅Π½ΠΎΠ±ΠΈΠΎΡΠΈΠΊΠΎΠ² ΠΌΠΈΠΊΡΠΎΠ±Π½ΡΠΌΠΈ ΡΠ΅ΡΠΌΠ΅Π½ΡΠ°ΠΌΠΈ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ° Π΄ΠΎ Π½Π΅ΠΊΠΎΡΠΎΡΠΎΠΉ ΡΡΠ΅ΠΏΠ΅Π½ΠΈ ΠΈΠ·Π²Π΅ΡΡΠ΅Π½, ΠΈΠ½ΡΠΎΡΠΌΠ°ΡΠΈΡ ΠΎ ΠΊΠΎΠ½ΠΊΡΠ΅ΡΠ½ΠΎΠΉ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΠ΅, ΠΎΠΏΠΎΡΡΠ΅Π΄ΡΡΡΠ΅ΠΉ ΠΊΠ°ΠΆΠ΄ΡΡ ΠΌΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΡΠ΅ΡΠΊΡΡ ΡΠ΅Π°ΠΊΡΠΈΡ, Π²ΡΡ Π΅ΡΡ ΠΎΠ³ΡΠ°Π½ΠΈΡΠ΅Π½Π°, Π² ΠΏΠ΅ΡΠ²ΡΡ ΠΎΡΠ΅ΡΠ΅Π΄Ρ, ΠΎΡΡΡΡΡΡΠ²ΠΈΠ΅ΠΌ Π°Π΄Π΅ΠΊΠ²Π°ΡΠ½ΠΎΠΉ ΠΌΠΎΠ΄Π΅Π»ΠΈ ΠΌΠΈΠΊΡΠΎΠ±Π½ΠΎΠ³ΠΎ ΡΠΎΠΎΠ±ΡΠ΅ΡΡΠ²Π° ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ°, ΠΏΠΎΠ·Π²ΠΎΠ»ΡΡΡΠ΅ΠΉ Π½Π°ΠΊΠ°ΠΏΠ»ΠΈΠ²Π°ΡΡ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°Π»ΡΠ½ΡΠ΅ Π΄Π°Π½Π½ΡΠ΅ Π΄Π»Ρ ΠΏΠΎΡΡΡΠΎΠ΅Π½ΠΈΡ Π²ΡΡΠΈΡΠ»ΠΈΡΠ΅Π»ΡΠ½ΡΡ
ΠΌΠΎΠ΄Π΅Π»Π΅ΠΉ. Π‘Π΅Π³ΠΎΠ΄Π½Ρ Π² ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠΈ ΠΌΠ΅ΡΠ°Π±ΠΎΠ»ΠΈΠ·ΠΌΠ° Π»Π΅ΠΊΠ°ΡΡΡΠ²Π΅Π½Π½ΡΡ
ΡΡΠ΅Π΄ΡΡΠ² ΠΏΡΠΈΠΌΠ΅Π½ΡΡΡ ΠΌΠΈΠΊΡΠΎΡΠ»ΡΠΈΠ΄Π½ΡΠ΅ ΡΠΈΠΏΡ, Π½Π° ΠΊΠΎΡΠΎΡΡΡ
ΡΡΠ½ΠΊΡΠΈΠΈ ΡΠ°Π·Π»ΠΈΡΠ½ΡΡ
ΠΎΡΠ³Π°Π½ΠΎΠ² ΠΈ ΡΠΊΠ°Π½Π΅ΠΉ, ΡΠ°ΠΊΠΈΡ
ΠΊΠ°ΠΊ ΠΏΠ΅ΡΠ΅Π½Ρ, ΠΏΠΎΡΠΊΠΈ, Π»Π΅Π³ΠΊΠΈΠ΅ ΠΈ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊ, Π²ΠΎΡΠΏΡΠΎΠΈΠ·Π²ΠΎΠ΄ΡΡΡΡ Π² Π²ΠΈΠ΄Π΅ in vitro ΠΌΠΎΠ΄Π΅Π»Π΅ΠΉ Π² ΡΠΎΡΠΌΠ΅ 2D ΠΈ 3D ΠΊΠ»Π΅ΡΠΎΡΠ½ΡΡ
ΠΊΡΠ»ΡΡΡΡ. ΠΠΎΠΏΠΎΠ»Π½Π΅Π½ΠΈΠ΅ ΡΠ°ΠΊΠΈΡ
ΡΠΈΡΡΠ΅ΠΌ ΠΌΠΈΠΊΡΠΎΠ±Π½ΡΠΌ ΡΠΎΠΎΠ±ΡΠ΅ΡΡΠ²ΠΎΠΌ ΠΏΠΎΠ·Π²ΠΎΠ»ΠΈΡ ΠΌΠ°ΠΊΡΠΈΠΌΠ°Π»ΡΠ½ΠΎ ΠΏΡΠΈΠ±Π»ΠΈΠ·ΠΈΡΡΡΡ ΠΊ ΠΌΠΎΠ΄Π΅Π»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ in vitro ΡΠ»ΠΎΠΆΠ½ΡΡ
Π±ΠΈΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΏΡΠΎΡΠ΅ΡΡΠΎΠ² Π² ΠΈΠ½ΡΠ΅ΡΠ΅ΡΠ°Ρ
ΡΠ°ΡΠΌΠ°ΠΊΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠΉ ΠΈ Π½Π°ΠΊΠΎΠΏΠ»Π΅Π½ΠΈΡ Π΄Π°Π½Π½ΡΡ
Π΄Π»Ρ ΠΏΠΎΡΡΡΠΎΠ΅Π½ΠΈΡ Π²ΡΡΠΈΡΠ»ΠΈΡΠ΅Π»ΡΠ½ΡΡ
ΠΌΠΎΠ΄Π΅Π»Π΅ΠΉ
Discrimination between Streptococcus pneumoniae and Streptococcus mitis based on sorting of their MALDI mass spectra
AbstractAccurate species-level identification of alpha-hemolytic (viridans) streptococci (VGS) is very important for understanding their pathogenicity and virulence. However, an extremely high level of similarity between VGS within the mitis group (S. pneumoniae, S. mitis, S. oralis and S. pseudopneumoniae) often results in misidentification of these organisms. Earlier, matrix-assisted laser desorption ionizationβtime of flight mass spectrometry (MALDI-TOF MS) has been suggested as a tool for the rapid identification of S. pneumoniae. However, by using Biotyper 3.0 (Bruker) or Vitek MS (bioMΓ©rieux) databases, Streptococcus mitis/oralis species can be erroneously identified as S. pneumoniae. ClinProTools 2.1 software was used for the discrimination of MALDI-TOF mass spectra of 25 S. pneumoniae isolates, 34 S. mitis and three S. oralis. Phenotypical tests and multilocus gene typing schemes for the S. pneumoniae (http://spneumoniae.mlst.net/) and viridans streptococci (http://viridans.emlsa.net/) were used for the identification of isolates included in the study. The classifying model was generated based on different algorithms (Genetic Algorithm, Supervised Neural Network and QuickClassifier). In all cases, values of sensitivity and specificity were found to be equal or close to 100%, allowing discrimination of mass spectra of different species. Three peaks (6949, 9876 and 9975 m/z) were determined conferring the maximal statistical weight onto each model built. We find this approach to be promising for viridans streptococci discrimination
ΠΠ·ΡΡΠ΅Π½ΠΈΠ΅ Π²ΠΈΠ΄ΠΎΠ²ΠΎΠ³ΠΎ ΡΠ°Π·Π½ΠΎΠΎΠ±ΡΠ°Π·ΠΈΡ Π±Π°ΠΊΡΠ΅ΡΠΈΠΉ ΡΠΎΠ΄Π° Bifidobacterium ΠΊΠΈΡΠ΅ΡΠ½ΠΎΠΉ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΡ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΠΌΠ΅ΡΠΎΠ΄Π° MALDI-TOF ΠΌΠ°ΡΡ-ΡΠΏΠ΅ΠΊΡΡΠΎΠΌΠ΅ΡΡΠΈΠΈ
Background: The members of genus Bifidobacterium represent a significant part of intestinal microbiota in adults and predominate in infants. Species repertoire of the intestinal bifidobacteria is known to be subjected to major changes with age; however, many details of this process are still to be elucidated.Objective: Our aim was to study the diversity of intestinal bifidobacteria and changes of their qualitative and quantitative composition characteristics during the process of growing up using MALDI-TOF mass-spectrometric analysis of pure bacterial cultures.Methods: A cross-sectional study of bifidobacteria in the intestinal microbiota was performed in 93 healthy people of the ages from 1 month to 57 years. Strains were identified using Microflex LT MALDI-TOF MS, the confirmation was performed by 16S rRNA gene fragment sequencing.Results: 93% of isolated bifidobacterial strains were successfully identified using MALDI-TOF mass-spectrometry. At least two of the strains from each species were additionally identified by 16S rRNA gene fragment sequencing, in all of the cases the results were the same. It was shown that the total concentration of bifidobacteria decreases with age (p 0.001) as well as the frequency of isolation of Bifidobacterium bifidum (p =0.020) and Bifidobacterium breve (p 0.001), and the frequency of isolation of Bifidobacterium adolescentis, increases (p 0.001), representing the continuous process of transformation of microbiota.Conclusion: The method of MALDI-TOF mass spectrometry demonstrated the ability to perform rapid and reliable identification of bifidobacteria that allowed the study of changes in the quantitative and qualitative characteristics of human microbiota in the process of growing up.ΠΡΠ΅Π΄ΡΡΠ°Π²ΠΈΡΠ΅Π»ΠΈ ΡΠΎΠ΄Π° Bifidobacterium ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»ΡΡΡ Π·Π½Π°ΡΠΈΡΠ΅Π»ΡΠ½ΡΡ ΡΠ°ΡΡΡ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΡ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ° Π²Π·ΡΠΎΡΠ»ΡΡ
Π»ΡΠ΄Π΅ΠΉ ΠΈ ΡΠΈΡΠ»Π΅Π½Π½ΠΎ Π΄ΠΎΠΌΠΈΠ½ΠΈΡΡΡΡ Π² ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΠ΅ ΠΌΠ»Π°Π΄Π΅Π½ΡΠ΅Π². ΠΠ·Π²Π΅ΡΡΠ½ΠΎ, ΡΡΠΎ Π²ΠΈΠ΄ΠΎΠ²ΠΎΠΉ ΡΠΎΡΡΠ°Π² ΠΊΠΈΡΠ΅ΡΠ½ΡΡ
Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ ΠΏΠΎΠ΄Π²Π΅ΡΠ³Π°Π΅ΡΡΡ ΡΠΈΠ»ΡΠ½ΡΠΌ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡΠΌ Ρ Π²ΠΎΠ·ΡΠ°ΡΡΠΎΠΌ, ΠΎΠ΄Π½Π°ΠΊΠΎ ΠΌΠ½ΠΎΠ³ΠΈΠ΅ Π΄Π΅ΡΠ°Π»ΠΈ ΡΡΠΎΠ³ΠΎ ΠΏΡΠΎΡΠ΅ΡΡΠ° ΠΎΡΡΠ°ΡΡΡΡ Π½Π΅ΡΡΠ½ΡΠΌΠΈ.Π¦Π΅Π»Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ: ΠΈΠ·ΡΡΠΈΡΡ Π²ΠΈΠ΄ΠΎΠ²ΠΎΠ΅ ΡΠ°Π·Π½ΠΎΠΎΠ±ΡΠ°Π·ΠΈΠ΅ Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ° ΠΈ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ ΠΈΡ
ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠ³ΠΎ ΠΈ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠ³ΠΎ ΡΠΎΡΡΠ°Π²Π° Π² ΠΏΡΠΎΡΠ΅ΡΡΠ΅ Π²Π·ΡΠΎΡΠ»Π΅Π½ΠΈΡ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° ΠΏΡΠΈ ΠΏΠΎΠΌΠΎΡΠΈ ΡΠ΅Ρ
Π½ΠΎΠ»ΠΎΠ³ΠΈΠΈ MALDI-TOF ΠΌΠ°ΡΡ-ΡΠΏΠ΅ΠΊΡΡΠΎΠΌΠ΅ΡΡΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ Π°Π½Π°Π»ΠΈΠ·Π° Π±Π΅Π»ΠΊΠΎΠ²ΡΡ
ΠΏΡΠΎΡΠΈΠ»Π΅ΠΉ ΡΠΈΡΡΡΡ
ΠΊΡΠ»ΡΡΡΡ.ΠΠ΅ΡΠΎΠ΄Ρ: ΠΊΡΠΎΡΡ-ΡΠ΅ΠΊΡΠΈΠΎΠ½Π½ΠΎΠ΅ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅ ΡΠ°Π·Π½ΠΎΠΎΠ±ΡΠ°Π·ΠΈΡ Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ Π² ΡΠΎΡΡΠ°Π²Π΅ Π½ΠΎΡΠΌΠ°Π»ΡΠ½ΠΎΠΉ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΡ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊΠ° ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½ΠΎ Ρ 93 ΡΠ΅Π»ΠΎΠ²Π΅ΠΊ Π² Π²ΠΎΠ·ΡΠ°ΡΡΠ΅ ΠΎΡ 1 ΠΌΠ΅Ρ Π΄ΠΎ 57 Π»Π΅Ρ. ΠΡΡΡΠ΅ΡΡΠ²Π»ΡΠ»ΠΈ Π²ΡΠ΄Π΅Π»Π΅- Π½ΠΈΠ΅ ΡΠΈΡΡΡΡ
ΠΊΡΠ»ΡΡΡΡ ΠΈ ΠΈΡ
ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΠΊΠ°ΡΠΈΡ Π½Π° ΠΏΡΠΈΠ±ΠΎΡΠ΅ Microflex LT MALDI-TOF MS (Bruker Daltonics, ΠΠ΅ΡΠΌΠ°Π½ΠΈΡ), ΠΏΠΎΠ΄ΡΠ²Π΅ΡΠΆΠ΄Π΅Π½ΠΈΠ΅ ΡΠ΅Π°Π»ΠΈΠ·ΠΎΠ²ΡΠ²Π°Π»ΠΈ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΡΠ΅ΠΊΠ²Π΅Π½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠ° Π³Π΅Π½Π° 16S ΡΠ ΠΠ.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ: Ρ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ΠΌ MALDI-TOF ΠΌΠ°ΡΡ-ΡΠΏΠ΅ΠΊΡΡΠΎΠΌΠ΅ΡΡΠΈΠΈ Π±ΡΠ»ΠΎ ΡΡΠΏΠ΅ΡΠ½ΠΎ ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΡΠΈΡΠΎΠ²Π°Π½ΠΎ 93% Π²ΡΠ΄Π΅Π»Π΅Π½Π½ΡΡ
ΡΡΠ°ΠΌΠΌΠΎΠ² Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ. ΠΠΈΠ½ΠΈΠΌΡΠΌ ΠΏΠΎ 2 ΠΏΡΠ΅Π΄ΡΡΠ°Π²ΠΈΡΠ΅Π»Ρ ΠΎΡ ΠΊΠ°ΠΆΠ΄ΠΎΠ³ΠΎ ΠΈΠ· Π²ΠΈΠ΄ΠΎΠ² Π±ΡΠ»ΠΈ Π΄ΠΎΠΏΠΎΠ»Π½ΠΈΡΠ΅Π»ΡΠ½ΠΎ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½Ρ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΡΠ΅ΠΊΠ²Π΅Π½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠ° Π³Π΅Π½Π° 16SΡΠ ΠΠ; Π²ΠΎ Π²ΡΠ΅Ρ
ΡΠ»ΡΡΠ°ΡΡ
ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΡ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠ² ΡΠΎΠ²ΠΏΠ°Π»ΠΈ. ΠΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ Ρ Π²ΠΎΠ·ΡΠ°ΡΡΠΎΠΌ ΠΏΡΠΎΠΈΡΡ
ΠΎΠ΄ΠΈΡ ΡΠ½ΠΈΠΆΠ΅Π½ΠΈΠ΅ ΠΎΠ±ΡΠ΅ΠΉ ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ (p 0,001), ΡΠΌΠ΅Π½ΡΡΠ°Π΅ΡΡΡ Π²ΡΡΡΠ΅ΡΠ°Π΅ΠΌΠΎΡΡΡ Π²ΠΈΠ΄ΠΎΠ² Bifidobacterium bifidum (p =0,020) ΠΈ Bifidobacterium breve (p 0,001), Π° Π²ΡΡΡΠ΅ΡΠ°Π΅ΠΌΠΎΡΡΡ Π²ΠΈΠ΄Π° Bifidobacterium adolescentis ΡΠ²Π΅Π»ΠΈΡΠΈΠ²Π°Π΅ΡΡΡ (p 0,001), ΠΎΡΡΠ°ΠΆΠ°Ρ ΠΏΠΎΡΡΠ΅ΠΏΠ΅Π½Π½ΡΠ΅ ΠΏΡΠΎΡΠ΅ΡΡΡ ΠΏΠ΅ΡΠ΅ΡΡΡΠΎΠΉΠΊΠΈ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΡ.ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅: ΠΌΠ΅ΡΠΎΠ΄ MALDI-TOF ΠΌΠ°ΡΡ-ΡΠΏΠ΅ΠΊΡΡΠΎΠΌΠ΅ΡΡΠΈΠΈ ΠΏΠΎΠΊΠ°Π·Π°Π» Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΡ Π±ΡΡΡΡΠΎΠΉ ΠΈ Π½Π°Π΄Π΅ΠΆΠ½ΠΎΠΉ ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ Π±ΠΈΡΠΈΠ΄ΠΎΠ±Π°ΠΊΡΠ΅ΡΠΈΠΉ, ΠΏΠΎΠ·Π²ΠΎΠ»ΠΈΠ²ΡΠ΅ΠΉ ΠΏΡΠΎΠ²Π΅ΡΡΠΈ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΠΉ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΡΡ
ΠΈ ΠΊΠ°ΡΠ΅ΡΡΠ²Π΅Π½Π½ΡΡ
ΠΏΠΎΠΊΠ°Π·Π°ΡΠ΅Π»Π΅ΠΉ ΠΌΠΈΠΊΡΠΎΡΠ»ΠΎΡΡ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° Π² ΠΏΡΠΎΡΠ΅ΡΡΠ΅ Π²Π·ΡΠΎΡΠ»Π΅Π½ΠΈ
GENERATION AND CHARACTERIZATION OF A NEUTRALIZING MONOCLONAL ANTIBODY AGAINST RABIES VIRUS
Rabies is a zoonotic disease, for which effective treatment methods after the onset of clinical symptoms have not been developed yet. Polyclonal sera, both human and equine, along with vaccines are important means of disease prophylaxis. However, due to adverse reactions to the immunoglobulins of animal origin, high cost, and limited availability of the safer human serum, polyclonal antibodies should be substituted for a stable and efficient preparation, which is recombinant neutralizing antirabies monoclonal antibodies (mAbs). This paper reports generation of the humanized mAb 1C5, which binds with the antigenic site (AS) III of the rabies virus glycoprotein (RABVG) and demonstrates high virus neutralization activity in the fluorescent antibody virus neutralization test, as a result of expression in the Chinese hamster ovary (CHO) cells
An improved and extended dual-index multiplexed 16S rRNA sequencing for the Illumina HiSeq and MiSeq platform
Abstract Background Recent advancements in next-generation sequencing (NGS) technology have ushered in significant improvements in sequencing speed and data throughput, thereby enabling the simultaneous analysis of a greater number of samples within a single sequencing run. This technology has proven particularly valuable in the context of microbial community profiling, offering a powerful tool for characterizing the microbial composition at the species level within a given sample. This profiling process typically involves the sequencing of 16S ribosomal RNA (rRNA) gene fragments. By scaling up the analysis to accommodate a substantial number of samples, sometimes as many as 2,000, it becomes possible to achieve cost-efficiency and minimize the introduction of potential batch effects. Our study was designed with the primary objective of devising an approach capable of facilitating the comprehensive analysis of 1,711 samples sourced from diverse origins, including oropharyngeal swabs, mouth cavity swabs, dental swabs, and human fecal samples. This analysis was based on data obtained from 16S rRNA metagenomic sequencing conducted on the Illumina MiSeq and HiSeq sequencing platforms. Results We have designed a custom set of 10-base pair indices specifically tailored for the preparation of libraries from amplicons derived from the V3-V4 region of the 16S rRNA gene. These indices are instrumental in the analysis of the microbial composition in clinical samples through sequencing on the Illumina MiSeq and HiSeq platforms. The utilization of our custom index set enables the consolidation of a significant number of libraries, enabling the efficient sequencing of these libraries in a single run. Conclusions The unique array of 10-base pair indices that we have developed, in conjunction with our sequencing methodology, will prove highly valuable to laboratories engaged in sequencing on Illumina platforms or utilizing Illumina-compatible kits
Gene Networks Underlying the Resistance of Bifidobacterium longum to Inflammatory Factors
As permanent residents of the normal gut microbiota, bifidobacteria have evolved to adapt to the hostβs immune response whose priority is to eliminate pathogenic agents. The mechanisms that ensure the survival of commensals during inflammation and maintain the stability of the core component of the normal gut microbiota in such conditions remain poorly understood.Β We propose a new in vitro approach to study the mechanisms of resistance to immune response factors based on high-throughput sequencing followed by transcriptome analysis. This approach allowed us to detect differentially expressed genes associated with inflammation. In this study, we demonstrated that the presence of the pro-inflammatory cytokines IL-6 and TNFΞ± to the growth medium of the B. longum subsp. longum GT15 strain changes the latterβs growth rate insignificantly while affecting the expression of certain genes. We identified these genes and performed a COG and a KEGG pathway enrichment analysis.Β Using phylogenetic profilingΒ we predicted the operons of genes whose expression was triggered by the cytokines TNFΞ± and IL-6 in vitro. By mapping the transcription start points, we experimentally validated the predicted operons. Thus, in this study, we predicted the genes involved in a putative signaling pathway underlying the mechanisms of resistance to inflammatory factors in bifidobacteria. Since bifidobacteria are a major component of the human intestinal microbiota exhibiting pronounced anti-inflammatory properties, this study is ofΒ great practical and scientific relevance. Β© Copyright Β© 2020 Veselovsky, Dyachkova, Menyaylo, Polyaeva, Olekhnovich, Shitikov, Bespiatykh, Semashko, Kasianov, Ilina, Danilenko and Klimina