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1.
Ann Lab Med ; 42(3): 358-362, 2022 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-34907106

RESUMEN

Identifying Mycobacterium using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is challenging. We evaluated the performance of MALDI-TOF MS in identifying nontuberculous mycobacteria (NTM) using the ASTA MicroIDSys system (ASTA Inc., Suwon, Korea) with the MycoDB v1.95s and upgraded MycoDB v2.0-beta databases. We tested 124 NTM isolates collected from Ogawa medium at Severance Hospital, Seoul, Korea, between January and April 2019. MicroIDSys scores were categorized into three groups: ≥140, reliable identification; 130-139, ambiguous identification; and <130, invalid identification. To validate the results, we used the reverse blot hybridization assay (Molecutech REBA MycoID, YD Diagnostics Corp., Korea). Initial analysis using MycoDB v1.95s resulted in 26.6% (33/124) reliable, 43.5% (54/124) ambiguous, and 29.8% (37/124) invalid identifications. Re-analysis using the upgraded MycoDB v2.0-beta database resulted in 94.4% (117/124) reliable, 4.0% (5/124) ambiguous, and 1.6% invalid (2/124) identifications. The percentage of reliable identifications that matched with the reference increased from 26.6% (33/124) with MycoDB v1.95s to 93.5% (116/124) with MycoDB v2.0-beta. The upgraded databases enable substantially improved NTM identification through deep learning in the inference algorithm and by considering more axes in the correlation analysis. MALDI-TOF MS using the upgraded database unambiguously identified most NTM species. Our study lays a foundation for applying MALDI-TOF MS for the simple and rapid identification of NTM isolated from solid media.


Asunto(s)
Mycobacterium , Micobacterias no Tuberculosas , Medios de Cultivo , Humanos , Rayos Láser , Micobacterias no Tuberculosas/genética , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
2.
Food Microbiol ; 102: 103924, 2022 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-34809950

RESUMEN

Exopolysaccharides production by 3 ropy strains of Lactobacillus delbrueckii subsp. bulgaricus of dairy origin was evaluated in synthetic medium by combining different approaches: impedometric measurements, fluorescent microscopy and flow cytometry analyses. The evaluation of ΔE by impedometric measurement (E%max-E%40h) allowed the detection of EPS production in synthetic medium, but the differences in EPS production kinetic was highlighted by flow cytometry analysis and fluorescent microcopy. This approach enabled us to unravel the diversity in EPS synthesis and release into the laboratory medium during the growth of the strains. Our results showed that the maximum EPS production occurred after 8 h of incubation, when cells were in late exponential growth phase. Furthermore, flow cytometry analysis revealed that only part of the cell population could be identified as EPS producer or as EPS-bounded cell. Therefore, the combined approach used, allowed us to define at the same time the kinetics of EPS production and release by three strains belonging to the same species and, highlight that the production of EPS depends also on the number of EPS-producing cells within the same population. This approach could be useful for the selection of strains to be used as starter cultures in dairy products where EPS production is considered an important feature.


Asunto(s)
Lactobacillus delbrueckii , Polisacáridos Bacterianos/biosíntesis , Medios de Cultivo , Productos Lácteos/microbiología , Fermentación , Lactobacillus delbrueckii/clasificación , Lactobacillus delbrueckii/metabolismo
3.
Bioresour Technol ; 344(Pt B): 126320, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-34780906

RESUMEN

A series of commercial powdered media (Cell-Hi F2P, JWP and WP) and a hydroponics medium (FloraMicroBloom) were investigated for the cultivation of P. tricornutum, and compared with f/2 (a commonly employed laboratory cultivation medium; costlier to scale). Cell-Hi JWP showed good performance characteristics including cost-effectiveness. Outdoor cultivation of P. tricornutum in an airlift photobioreactor, using Cell-Hi JWP in the United Kingdom (UK) during September and October (average daily temperature ranging between 8 and 18 °C and natural sunlight) was comparable to cultivation indoors under controlled temperature and lighting. A strong positive correlation between fucoxanthin and chlorophyll a content, and a weak inverse correlation between eicosapentaenoic (EPA) content and temperature were observed. Commensal bacterial counts revealed a sinusoidal growth profile with a change in community dominance from Halomonas sp. to Marinobacter sp. This investigation reveals for the first time that a multi-product approach can be adopted with P. tricornutum in a UK outdoor environment using commercially viable powdered media.


Asunto(s)
Diatomeas , Microalgas , Clorofila A , Medios de Cultivo , Fotobiorreactores , Reino Unido
4.
Bioresour Technol ; 344(Pt B): 126356, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-34822989

RESUMEN

Using municipal wastewater sludge to produce microbial lipid is an effective way of resource recycling. Sludge contains heavy metals and may lead to negative impact on lipid production. However, relative study has not been reported. In this study, metal impact on Lipomyces starkeyi lipid accumulation was conducted. Results showed that Cd2+ had great impact on lipid accumulation, but other metals had no much impact. The maximum lipid content of L. starkeyi cultivated in 0.55 mg/L of Cd2+ was only 41% w/w, which was lower than the control (51% w/w). The inhibition on acetyl-CoA formation was observed when Cd2+ was in the medium. After removing metals from sludge, the lipid accumulation was only around half of the one without metal removal. It would be due to that not only the toxic metals in the sludge were removed as well as the metals such as Zn2+ which can enhance lipid accumulation.


Asunto(s)
Metales Pesados , Aguas del Alcantarillado , Medios de Cultivo , Lípidos , Aguas Residuales
5.
Methods Mol Biol ; 2373: 177-199, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-34520013

RESUMEN

Pumpless microfluidic systems are easy-to-use devices that can be used to culture cells that are sensitive to mechanical shear, such as lymphatic endothelial cells. However, previously developed pumpless systems either provide unidirectional shear where the cell culture medium is discarded, or bidirectional shear that produces endothelial cell cultures with disease characteristics. Here, we describe a PDMS-based system that produces cyclically rising and falling shear that is unidirectional, similar to what has been reported in lymphatic vessels. The system can recirculate cell culture medium, making it possible for proteins and growth factors produced by the cell culture to remain in circulation. In addition, we describe the custom-made rotating platform that we used to create this unique flow pattern. Using this rotating platform, the microfluidic device can be used to grow confluent layers of lymphatic endothelial cells under physiologically relevant growth conditions.


Asunto(s)
Vasos Linfáticos , Microfluídica , Técnicas de Cultivo de Célula , Medios de Cultivo , Células Endoteliales , Dispositivos Laboratorio en un Chip
6.
Int J Food Microbiol ; 362: 109497, 2022 Feb 02.
Artículo en Inglés | MEDLINE | ID: mdl-34896913

RESUMEN

The filter concentration method facilitates the rapid detection of foodborne pathogens. The filter concentration method lowered the limit of detection (LOD) of artificially inoculated cabbage with Salmonella Typhimurium; however, the procedure injured foodborne pathogens during filtering procedure. Thus, to detect injured pathogens under the detection limit, an enrichment broth promoting pathogen resuscitation and growth is required. To rapidly recover, cultivate and lower the time to result (TTR) of S. Typhimurium detection after filter concentration method, a brain heart infusion (BHI) broth-based modified enrichment broth (MEB) was developed. The MEB was developed by fitting growth curves to a modified Gompertz model; 1.00 g/L of sodium pyruvate, 0.20 g/L proline and 2.0 g/L magnesium sulphate additives were optimized as additional components to rapidly grow filter-injured S. Typhimurium. As a result, the rate of filter-injured S. Typhimurium went from 100% to 0.0% using MEB within 3.5 h. In contrast, BHI required 4 h and buffered peptone water (BPW) required more than 4 h to decrease the injury rate to 0.0%. Using MEB, BHI and BPW, filter-injured S. Typhimurium in cabbages were enriched to 4.056 ± 0.026 Log CFU/25 g, 3.571 ± 0.187 Log CFU/25 g and 3.708 ± 0.156 Log CFU/25 g, respectively. Additionally, 1-9 CFU/mL S. Typhimurium in cabbage was detected within 3.0 h, including 1 h enrichment with MEB, whereas 5.0 h was required for BHI and BPW. Thus, the MEB developed in this study showed great potential as a short enrichment broth for the rapid detection of filter-injured S. Typhimurium.


Asunto(s)
Brassica , Salmonella typhimurium , Medios de Cultivo , Microbiología de Alimentos
7.
Gene ; 809: 146012, 2022 Jan 30.
Artículo en Inglés | MEDLINE | ID: mdl-34655719

RESUMEN

Cancer cells rewire metabolic pathways as they demand more ATP and building blocks for proliferation. Glucose is the most consumed nutrient by cancer cells and metabolized to lactate even in the presence of oxygen. This phenomenon is called 'aerobic glycolysis'. Also, glucose level is found lower in tumor environment. Leukemia is characterized by abnormal proliferation of hematopoietic cells. STAT3 a transcription factor and an oncogene is upregulated in many tumor types. Despite its well-defined functions, STAT3 has also been proposed as a metabolic regulator. In this study, we aimed to determine the role STAT3 activation in glucose limitation, in leukemia cell lines. K562, NB-4 and HL-60 cells were found sensitive to glucose limitation. In low glucose conditions, total and nuclear STAT3 protein was decreased in all cells. In mitochondria, S727 phosphorylated STAT3 (mitochondrial form) was determined slightly increased in K562 and NB-4 cells. On the other side, ectopically STAT3 expressing cells had increased glucose consumption and less proliferated in low glucose medium. This data suggests that aerobic glycolysis might be upregulated upon STAT3 expression in leukemia cells, in glucose limitation. Furthermore, in this study, it was found that GLUT3 expressing cells did not reduce STAT3 expression in low glucose medium. GLUT3 was previously determined as a molecular marker for cell sensitivity to glucose limitation, therefore, it could be hypothesized as GLUT3 expressing cells might not need to alter STAT3 expression in low glucose level. Overall, our data suggest that leukemia cells rewire glucose metabolism via STAT3 expression in glucose limitation. Elucidating pathways that cause differential phosphorylation of STAT3 and its interaction with other energy regulating pathways in cellular response to glucose limitation might be beneficial to design new drug targets such as STAT3 inhibitors for leukemia treatment.


Asunto(s)
Glucosa/metabolismo , Leucemia/metabolismo , Factor de Transcripción STAT3/metabolismo , Línea Celular Tumoral , Proliferación Celular , Medios de Cultivo/química , Regulación hacia Abajo , Regulación Leucémica de la Expresión Génica , Transportador de Glucosa de Tipo 3/metabolismo , Glucólisis/fisiología , Células HL-60 , Humanos , Células K562 , Leucemia/genética , Leucemia/patología , Mitocondrias/metabolismo , Factor de Transcripción STAT3/genética
8.
Methods Mol Biol ; 2391: 55-62, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-34686976

RESUMEN

The appearance of colony growth sectors on solid medium plates has been described in many fungi. Although the molecular bases of this phenomenon remain largely unknown, possible relationships with genetic or epigenetic changes have been reported. Here we present a method to quantify the frequency of colony growth sectors in Fusarium oxysporum, which can be used to compare different fungal strains and to infer their genetic instability.


Asunto(s)
Fusarium/genética , Medios de Cultivo
9.
Methods Mol Biol ; 2387: 17-28, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-34643898

RESUMEN

Primary isolation of Mycobacterium ulcerans is the separation and growth of the bacterium from a mixed population either in clinical specimen or environmental specimen in pure cultures. It is a crucial activity as it can be used to monitor antimicrobial treatment, surveillance for antimicrobial resistance, and molecular epidemiology studies toward understanding pathogen ecology and transmission as well as pathogen biology. The process involves removal of unwanted fast-growing bacteria using 5% oxalic acid, inoculation on Lowenstein-Jensen medium supplemented with glycerol, and incubation at temperatures between 30 °C and 33 °C.


Asunto(s)
Mycobacterium ulcerans , Antiinfecciosos , Medios de Cultivo , Glicerol
10.
Braz J Biol ; 84: e253514, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34932678

RESUMEN

The objective of the current study was to investigate the synergistic impact of α-Tocopherol and α-Linolenic acid (100 µM) on IVM and IVC of Nili Ravi buffalo oocytes. Oocytes were obtained from the ovaries of slaughtered buffaloes within two hours after slaughter and brought to laboratory. Buffalo cumulus oocyte complexes were placed randomly in the five experimental groups included; GROUP 1: Maturation media (MM) + 100 µM ALA (control), GROUP 2: MM + 100 µM ALA + 50µM α-Tocopherol, GROUP 3: MM + 100 µM ALA + 100µM α-Tocopherol, GROUP 4: MM + 100 µM ALA + 200 µM α-Tocopherol and GROUP 5: MM + 100 µM ALA + 300 µM α-Tocopherol under an atmosphere of 5% CO2 in air at 38.5 °C for 22-24 h. Cumulus expansion and nuclear maturation status was determined (Experiment 1). In experiment 2, oocytes were matured as in experiment 1. The matured oocytes were then fertilized in Tyrode's Albumin Lactate Pyruvate (TALP) medium for about 20 h and cultured in synthetic oviductal fluid (SOF) medium to determine effect of α-Linolenic acid (100 µM) and α-Tocopherol in IVM medium on IVC of presumptive zygotes. To study the effect of α-Linolenic acid (100 µM) in IVM media and increasing concentration of α-tocopherol in the culture media on early embryo development (Experiment 3), the presumptive zygotes were randomly distributed into the five experimental groups with increasing concentration of α-tocopherol in culture media. Higher percentage of MII stage oocytes in experiment 1(65.2±2.0), embryos at morula stage in experiment 2 (30.4±1.5) and experiment 3 (22.2±2.0) were obtained. However, overall results for cumulus cell expansion, maturation of oocyte to MII stage and subsequent embryo development among treatments remain statistically similar (P > 0.05). Supplementation of α-tocopherol in maturation media having α-Linolenic acid and/or in embryo culture media did not further enhance in vitro maturation of oocyte or embryo production.


Asunto(s)
Búfalos , Ácido alfa-Linolénico , Animales , Medios de Cultivo , Desarrollo Embrionario , Oocitos , Ácido alfa-Linolénico/farmacología , alfa-Tocoferol/farmacología
11.
Ann Parasitol ; 67(3): 445-453, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34953119

RESUMEN

Hydatidosis is a disease caused by the larval stage of Echinococcus granulosus, which has great importance in medicine and veterinary medicine. Protoscolices (PSCs) of fertile hydatid cysts play a critical role in secondary echinococcosis after surgery. Fertile cysts were acquired from infected sheep at the local municipal abattoir in Ahvaz, southwest of Iran. PSCs were obtained aseptically and transferred to 10 different culture media and kept at 4°C and 37°C to determine the duration of PSCs' survival. Then, 2000 live PSCs from each of the culture media were injected into the peritoneal cavity of BALB/c mice. After five months, the mice were evaluated in terms of cyst number, size, and weight. The highest PSCs survival time at 4°C was related to RPMI-1640 medium and cyst fluid for 50 and 45 days, respectively. Also, at 37°C, the longest survival time of PSCs was related to cyst fluid and RPMI-1640 media for 30 and 29 days, respectively. The highest level of infection and median cyst number was observed in mice received PSCs from the RPMI-1640 medium at 4°C, and the highest level of infection in mice at 37°C was related to the DMEM low glucose (L) medium. The current study indicated that 4°C was a more suitable temperature for in vitro storage of live PSCs. The maximum amount of infection was observed in mice received PSCs from the RPMI-1640 medium at 4°C. The present study is the first attempt to compare the ability of PSCs to generate hydatid cysts in mice after being cultured in different media and at various temperatures.


Asunto(s)
Quistes , Echinococcus granulosus , Echinococcus , Animales , Medios de Cultivo , Ratones , Ovinos , Temperatura
12.
PLoS One ; 16(12): e0261191, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34928974

RESUMEN

The intestinal mucus layer plays a crucial role in human health. To study intestinal mucus function and structure in vitro, the mucus-producing intestinal cell line HT29-MTX-E12 has been commonly used. However, this cell line produces only low amounts of the intestine-specific MUC2. It has been shown previously that HT29-MTX-E12 cells cultured under Semi-Wet interface with Mechanical Stimulation (SWMS) produced higher amounts of MUC2, concomitant with a thicker mucus layer, compared to cells cultured conventionally. However, it remains unknown which underlying pathways are involved. Therefore, we aimed to further explore the cellular processes underlying the increased MUC2 production by HT29-MTX-E12 cells grown under SWMS conditions. Cells grown on Transwell membranes for 14 days under static and SWMS conditions (after cell seeding and attachment) were subjected to transcriptome analysis to investigate underlying molecular pathways at gene expression level. Caco-2 and LS174T cell lines were included as references. We characterized how SWMS conditions affected HT29-MTX-E12 cells in terms of epithelial barrier integrity, by measuring transepithelial electrical resistance, and cell metabolism, by monitoring pH and lactate production per molecule glucose of the conditioned medium. We confirmed higher MUC2 production under SWMS conditions at gene and protein level and demonstrated that this culturing method primarily stimulated cell growth. In addition, we also found evidence for a more aerobic cell metabolism under SWMS, as shown previously for similar models. In summary, we suggest different mechanisms by which MUC2 production is enhanced under SWMS and propose potential applications of this model in future studies.


Asunto(s)
Biomarcadores de Tumor/metabolismo , Técnicas de Cultivo de Célula/métodos , Neoplasias del Colon/patología , Regulación Neoplásica de la Expresión Génica , Moco/metabolismo , Apoptosis , Biomarcadores de Tumor/genética , Fenómenos Biomecánicos , Células CACO-2 , Ciclo Celular , Proliferación Celular , Neoplasias del Colon/genética , Neoplasias del Colon/metabolismo , Medios de Cultivo , Células HT29 , Humanos
13.
Biomolecules ; 11(12)2021 12 20.
Artículo en Inglés | MEDLINE | ID: mdl-34944556

RESUMEN

This study hypothesizes that bacteria inhabiting shale rock affect the content of the sedimentary cobalt protoporphyrin present in it and can use it as a precursor for heme synthesis. To verify this hypothesis, we conducted qualitative and quantitative comparative analyses of cobalt protoporphyrin as well as heme, and heme iron in shale rock that were (i) inhabited by bacteria in the field, (ii) treated with bacteria in the laboratory, and with (iii) bacterial culture on synthetic cobalt protoporphyrin. Additionally, we examined the above-mentioned samples for the presence of enzymes involved in the heme biosynthesis and uptake as well as hemoproteins. We found depletion of cobalt protoporphyrin and a much higher heme concentration in the shale rock inhabited by bacteria in the field as well as the shale rock treated with bacteria in the laboratory. Similarly, we observed the accumulation of protoporphyrin in bacterial cells grown on synthetic cobalt protoporphyrin. We detected numerous hemoproteins in metaproteome of bacteria inhabited shale rock in the field and in proteomes of bacteria inhabited shale rock and synthetic cobalt protoporhyrin in the laboratory, but none of them had all the enzymes involved in the heme biosynthesis. However, proteins responsible for heme uptake, ferrochelatase and sirohydrochlorin cobaltochelatase/sirohydrochlorin cobalt-lyase were detected in all studied samples.


Asunto(s)
Bacterias/crecimiento & desarrollo , Fósiles/microbiología , Sedimentos Geológicos/microbiología , Hemo/análisis , Protoporfirinas/análisis , Bacterias/metabolismo , Proteínas Bacterianas/metabolismo , Técnicas Bacteriológicas , Medios de Cultivo/química , Ferroquelatasa/metabolismo , Regulación Bacteriana de la Expresión Génica , Sedimentos Geológicos/química , Hemo/biosíntesis , Liasas/metabolismo , Proteómica , Protoporfirinas/biosíntesis
14.
Proc Natl Acad Sci U S A ; 118(51)2021 12 21.
Artículo en Inglés | MEDLINE | ID: mdl-34916298

RESUMEN

The thyroid maintains systemic homeostasis by regulating serum thyroid hormone concentrations. Here we report the establishment of three-dimensional (3D) organoids from adult thyroid tissue representing murine and human thyroid follicular cells (TFCs). The TFC organoids (TFCOs) harbor the complete machinery of hormone production as visualized by the presence of colloid in the lumen and by the presence of essential transporters and enzymes in the polarized epithelial cells that surround a central lumen. Both the established murine as human thyroid organoids express canonical thyroid markers PAX8 and NKX2.1, while the thyroid hormone precursor thyroglobulin is expressed at comparable levels to tissue. Single-cell RNA sequencing and transmission electron microscopy confirm that TFCOs phenocopy primary thyroid tissue. Thyroid hormones are readily detectable in conditioned medium of human TFCOs. We show clinically relevant responses (increased proliferation and hormone secretion) of human TFCOs toward a panel of Graves' disease patient sera, demonstrating that organoids can model human autoimmune disease.


Asunto(s)
Regulación de la Expresión Génica/fisiología , Enfermedad de Graves/metabolismo , Organoides/metabolismo , Células Epiteliales Tiroideas/fisiología , Animales , Medios de Cultivo , Humanos , Ratones , Factor de Transcripción PAX8/genética , Factor de Transcripción PAX8/metabolismo , Tiroglobulina/genética , Tiroglobulina/metabolismo , Factor Nuclear Tiroideo 1/genética , Factor Nuclear Tiroideo 1/metabolismo
15.
Arch Microbiol ; 204(1): 90, 2021 Dec 28.
Artículo en Inglés | MEDLINE | ID: mdl-34962612

RESUMEN

The choice of inoculum for successful isolation and establishment of axenic lichen mycobiont culture is a key step towards eliminating endolichenic and lichenicolous fungi and other microbial contamination. The nutritional requirements of each lichen species are unique. This work reports on the isolation, phenotypic plasticity, growth and secondary metabolites from mycobiont culture of the pantropical lichen Platygramme caesiopruinosa. Media composition [Malt yeast extract (MY), Modified Murashige and Skoog (MMS) and Lilly and Barnett (LB) media] was optimized to determine nutritional requirements for optimal growth of this species as assessed by dry biomass and the occurrence of secondary metabolite. Furthermore, the role of different carbon sources in affecting growth, growth stages, phenotypic plasticity, biomass and spectrum of secondary metabolites produced of this mycobiont was examined. The molecular identity of the mycobiont culture was confirmed by amplifying mitochondrial small subunit (mtSSU) sequences. Cultures showed optimum biomass production in MY medium with 10% sucrose. The secondary metabolite profiles for each culture treatment were characterized using High-performance Thin-Layer Chromatography (HPTLC) and Gas Chromatography with Mass Spectrometric (GC-MS) analysis. The HPTLC spectral comparison, phenolic and iodine confirmatory analysis revealed the absence of phenolic metabolites and the presence of non-phenolic metabolites in mycobiont extracts, while GC-MS analysis revealed the biosynthesis of side chain fatty acids, hydrocarbons and sugar alcohol in mycobiont cultures treated with 10% supplemented sucrose as a carbon source.


Asunto(s)
Ascomicetos , Líquenes , Adaptación Fisiológica , Medios de Cultivo
16.
Int J Mol Sci ; 22(24)2021 Dec 11.
Artículo en Inglés | MEDLINE | ID: mdl-34948135

RESUMEN

Brain pathologies evoked by thiamine deficiency can be aggravated by mild zinc excess. Cholinergic neurons are the most susceptible to such cytotoxic signals. Sub-toxic zinc excess aggravates the injury of neuronal SN56 cholinergic cells under mild thiamine deficiency. The excessive cell loss is caused by Zn interference with acetyl-CoA metabolism. The aim of this work was to investigate whether and how astroglial C6 cells alleviated the neurotoxicity of Zn to cultured SN56 cells in thiamine-deficient media. Low Zn concentrations did not affect astroglial C6 and primary glial cell viability in thiamine-deficient conditions. Additionally, parameters of energy metabolism were not significantly changed. Amprolium (a competitive inhibitor of thiamine uptake) augmented thiamine pyrophosphate deficits in cells, while co-treatment with Zn enhanced the toxic effect on acetyl-CoA metabolism. SN56 cholinergic neuronal cells were more susceptible to these combined insults than C6 and primary glial cells, which affected pyruvate dehydrogenase activity and the acetyl-CoA level. A co-culture of SN56 neurons with astroglial cells in thiamine-deficient medium eliminated Zn-evoked neuronal loss. These data indicate that astroglial cells protect neurons against Zn and thiamine deficiency neurotoxicity by preserving the acetyl-CoA level.


Asunto(s)
Neuronas Colinérgicas/metabolismo , Neuroglía/metabolismo , Deficiencia de Tiamina/prevención & control , Zinc/toxicidad , Animales , Línea Celular Tumoral , Medios de Cultivo , Ratones , Tiamina/metabolismo , Tiamina/farmacología , Deficiencia de Tiamina/metabolismo
17.
Georgian Med News ; (319): 124-128, 2021 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-34749336

RESUMEN

The scientific and practical significance of the problem of urogenital trichomoniasis is determined by the prevalence and variability of the pathogen. It is assumed that under adverse conditions, these protozoa lose the ability to move intensively and undergo morphological transformation. The existence of such a problem generates the scientific and practical interest in the matter of improvement of bacteriological methods of diagnosis when examining patients with chronic inflammatory processes of the urogenital tract. The purpose of the study - optimization of the composition of the nutrient medium for bacteriological detection of the trichomoniasis pathogen, considering the existence of different morphotypes and ways of trichomonas' motility. 50 culture samples of Trichomonas vaginalis, taken from 293 patients with chronic diseases of the genitourinary system, served as the object of study, in their comparative cultivation in the improved and standard environment using the methods of classical bacteriology. It has been evidentiated that the optimal ratio of additional ingredients, tested in the course of the experiments, has led to the increased growth and increased biomass of the pathogen, indicating the improvement in the quality of the nutrient medium. The data obtained indicates that the changed components of the optimized nutrient medium have improved the quality of the diagnostic procedure especially in patients with a torpid course of the disease.


Asunto(s)
Tricomoniasis , Trichomonas vaginalis , Técnicas Bacteriológicas , Medios de Cultivo , Humanos , Nutrientes , Tricomoniasis/diagnóstico
18.
Braz J Biol ; 83: e250550, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34730714

RESUMEN

Vanillin is the major component which is responsible for flavor and aroma of vanilla extract and is produced by 3 ways: natural extraction from vanilla plant, chemical synthesis and from microbial transformation. Current research was aimed to study bacterial production of vanillin from native natural sources including sewage and soil from industrial areas. The main objective was vanillin bio-production by isolating bacteria from these native sources. Also to adapt methodologies to improve vanillin production by optimized fermentation media and growth conditions. 47 soil and 13 sewage samples were collected from different industrial regions of Lahore, Gujranwala, Faisalabad and Kasur. 67.7% bacterial isolates produced vanillin and 32.3% were non-producers. From these 279 producers, 4 bacterial isolates selected as significant producers were; A3, A4, A7 and A10. These isolates were identified by ribotyping as A3 Pseudomonas fluorescence (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) and A10 Bacillus subtilis (KT962919). Vanillin producers were further tested for improved production of vanillin and were grown in different fermentation media under optimized growth conditions for enhanced production of vanillin. The fermentation media (FM) were; clove oil based, rice bran waste (residues oil) based, wheat bran based and modified isoeugenol based. In FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36, and FM37, the selected 4 bacterial strains produced significant amounts of vanillin. A10 B. subtilis produced maximum amount of vanillin. This strain produced 17.3 g/L vanillin in FM36. Cost of this fermentation medium 36 was 131.5 rupees/L. This fermentation medium was modified isoeugenol based medium with 1% of isoeugenol and 2.5 g/L soybean meal. ech gene was amplified in A3 P. fluorescence using ech specific primers. As vanillin use as flavor has increased tremendously, the bioproduction of vanillin must be focused.


Asunto(s)
Benzaldehídos , Aromatizantes , Alcaligenes faecalis/metabolismo , Bacillus subtilis/metabolismo , Benzaldehídos/metabolismo , Medios de Cultivo , Enterococcus faecium/metabolismo , Fermentación , Aromatizantes/metabolismo , Microbiología Industrial , Pseudomonas fluorescens/metabolismo
19.
Antonie Van Leeuwenhoek ; 114(12): 2047-2063, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-34609626

RESUMEN

The waste and by-products of the soybean industry could be an economic source of nutrients to satisfy the high nutritional demands for the cultivation of lactic acid bacteria. The aims of this work were to maximize the biomass production of Lacticaseibacillus paracasei 90 (L90) in three culture media formulated from an effluent derived from soy protein concentrate production and to assess the effects these media have on the enzymatic activity of L90, together with their influence on its fermentation profile in milk. The presence of essential minerals and fermentable carbohydrates (sucrose, raffinose, and stachyose) in the effluent was verified. L90 reached high levels of microbiological counts (∼ 9 log cfu mL-1) and dry weight (> 1 g L-1) on the three optimized media. Enzymatic activities (lactate dehydrogenase and ß-galactosidase) of L90, and its metabolism of lactose and citric acid, as well as lactic acid and pyruvic acid production in milk, were modified depending on the growth media. The ability of the L90 to produce the key flavour compounds (diacetyl and acetoin) was maintained or improved by growing in the optimized media in comparison with MRS.


Asunto(s)
Minerales , Proteínas de Soja , Biomasa , Medios de Cultivo , Fermentación
20.
Lab Chip ; 21(20): 3963-3978, 2021 10 12.
Artículo en Inglés | MEDLINE | ID: mdl-34636813

RESUMEN

Organ-on-chip (OoC) systems have become a promising tool for personalized medicine and drug development with advantages over conventional animal models and cell assays. However, the utility of OoCs in industrial settings is still limited, as external pumps and tubing for on-chip fluid transport are dependent on error-prone, manual handling. Here, we present an on-chip pump for OoC and Organ-Disc systems, to perfuse media without external pumps or tubing. Peristaltic pumping is implemented through periodic compression of a flexible pump layer. The disc-shaped, microfluidic module contains four independent systems, each lined with endothelial cells cultured under defined, peristaltic perfusion. Both cell viability and functionality were maintained over several days shown by supernatant analysis and immunostaining. Integrated, on-disc perfusion was further used for cytokine-induced cell activation with physiologic cell responses and for whole blood perfusion assays, both demonstrating the versatility of our system for OoC applications.


Asunto(s)
Células Endoteliales , Dispositivos Laboratorio en un Chip , Animales , Medios de Cultivo , Microfluídica , Perfusión
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