In vitro Cholesterol Assimilation by Bifidobacterium animalis subsp. lactis (BPL1) Probiotic Bacteria under Intestinal Conditions.

Author(s):  
María Coronada Fernández-Calderón ◽  
María Dolores Hinchado Sánchez-Moro ◽  
Eduardo Ortega Rincón

Background: Hypercholesterolemia is one of the principal causes of the development of cardiovascular diseases. Recently, probiotics consumption has been also proposed as a non-pharmacological intervention to control cholesterol concentrations. Objective: To evaluate in vitro assimilation of cholesterol by Bifidobacterium animalis subsp. lactis (BPL1) under simulated intestinal environment in anaerobic conditions and to review and discuss potential physiological mechanisms in this context. Methods: Bacterial viability and cholesterol assimilation was evaluated in both standard MRS and stimulated intestinal fluid (SIF) medium under anaerobic conditions, and in presence or absence of cholesterol. For assimilation assays, cholesterol concentrations in the different suspensions, containing the probiotic or not, was determined by chromatography coupled to mass spectrometry. Results: Results showed that the growth of B. lactis BPL1 under intestinal conditions is favored when cholesterol is present in the culture medium. In addition, a cholesterol assimilation of up to 44.4% under intestinal and anaerobic conditions was observed. Conclusion: Taking into account the revised literature and the experimental results herein presented, administration of functional foodstuffs together with probiotic bacteria such as B. lactis BPL1 could be a potential effective option to decrease hypercholesterolemia, thus preventing the development of cardiovascular diseases. Nevertheless, further studies on mechanisms of effectiveness in animals and clinical trials are still needed.

Blood ◽  
1967 ◽  
Vol 30 (6) ◽  
pp. 691-706 ◽  
Author(s):  
LAUREN NT. PACHMAN

Abstract 1. A homogeneous population of small lymphocytes with an average size of 6.7 micra was isolated from equine blood. 2. These cells could be maintained in vitro, with essentially complete survival for 24 hours, and with a 50% viability for five days. 3. The small lymphocytes consumed glucose at a rate of 1.87 mµmoles/1 x 107 cells/minute, and produced lactic acid at a rate of 2.30 mµmoles/1 x 107 cells/minute. 4. The oxygen consumption of small equine lymphocytes was 1.023 ± .165 mµmo1es oxygen 1 x 107 cells/minute, and that of mixed peripheral blood leukocytes, 1.25 ± .07 mµmoles oxygen/1 x 107 cells/minute, as determined, using a Clark oxygen electrode. 5. Lymphocyte glycolysis was stimulated under anaerobic conditions (Pasteur effect), and viability appeared unimpaired after 24 hours in a N2 environment. 6. Uncoupling of oxidative phosphorylation by the addition of 2,4, dinitrophenol stimulated both respiration and glycolysis. 7. The glycogen content of the normal small horse lymphocyte was 7.45 ± 1.04 µg glucose equivalents per 1 x 107 cells. 8. Many of the initial cell population were transformed into "blasts" following the addition of phytohemagglutin to the tissue culture medium. This response was associated with an increase in the rate of glycolysis and respiration by 24 hours, and a rise in intracellular glycogen by 48 hours.


Cells ◽  
2019 ◽  
Vol 8 (11) ◽  
pp. 1383 ◽  
Author(s):  
John Papaconstantinou

The ASK1-signalosome→p38 MAPK and SAPK/JNK signaling networks promote senescence (in vitro) and aging (in vivo, animal models and human cohorts) in response to oxidative stress and inflammation. These networks contribute to the promotion of age-associated cardiovascular diseases of oxidative stress and inflammation. Furthermore, their inhibition delays the onset of these cardiovascular diseases as well as senescence and aging. In this review we focus on whether the (a) ASK1-signalosome, a major center of distribution of reactive oxygen species (ROS)-mediated stress signals, plays a role in the promotion of cardiovascular diseases of oxidative stress and inflammation; (b) The ASK1-signalosome links ROS signals generated by dysfunctional mitochondrial electron transport chain complexes to the p38 MAPK stress response pathway; (c) the pathway contributes to the sensitivity and vulnerability of aged tissues to diseases of oxidative stress; and (d) the importance of inhibitors of these pathways to the development of cardioprotection and pharmaceutical interventions. We propose that the ASK1-signalosome regulates the progression of cardiovascular diseases. The resultant attenuation of the physiological characteristics of cardiomyopathies and aging by inhibition of the ASK1-signalosome network lends support to this conclusion. Importantly the ROS-mediated activation of the ASK1-signalosome p38 MAPK pathway suggests it is a major center of dissemination of the ROS signals that promote senescence, aging and cardiovascular diseases. Pharmacological intervention is, therefore, feasible through the continued identification of potent, non-toxic small molecule inhibitors of either ASK1 or p38 MAPK activity. This is a fruitful future approach to the attenuation of physiological aspects of mammalian cardiomyopathies and aging.


2017 ◽  
Vol 49 (02) ◽  
pp. 148-152
Author(s):  
Johannes Von den Hoff ◽  
Laury Fuentes ◽  
Marjon Bloemen ◽  
Anne Kuijpers-Jagtman

AbstractThe mouse is an excellent model to study palatogenesis since, like humans, it is a mammal, it can easily be bred and a wide array of molecular tools are available. Moreover, the isolated embryonic mouse palate can be cultured in vitro, which allows us to study the actual fusion process in detail.Just prior to the actual fusion at E13, the maxilla with the 2 palatal shelves can be dissected from the embryo head. The maxilla is cultured on a filter paper on top of a stainless-steel grid at the interface of the culture medium and air. After 24 h in culture, the palatal shelves are in contact and the MES is clearly visible. At 48 h, the MES has nearly completely disintegrated with small remnants at the nasal and oral side of the palate. Finally, at 72 h, the palatal shelves are completely fused and osteoid tissue is beginning to form in the lateral areas.The exact downstream molecular mechanisms that lead to clefting are far from being unraveled. The model for palate fusion presented here offers the possibility to analyze the molecular and cellular mechanisms that lead to clefting as a result of specific genetic and environmental factors. This will pave the way to pharmacological intervention for the prevention of cleft lip and/or palate in susceptible individuals.


1976 ◽  
Vol 4 (4) ◽  
pp. 360-371
Author(s):  
P L Sandok ◽  
S T Knight ◽  
H M Jenkin

Treponema pallidum (Nichols virulent) was incubated with and without cells in cell culture medium reduced to -275 mV Ecal, pH 7.3, under deoxygenated conditions. Five to ten percent of the treponemes attached to cells and remained motile for at least 120 h in cell-treponeme systems of co-incubation. Virulent treponemes could be detected after 120 to 144 h in the supernatant fluids of cell-treponeme co-incubation cultures and in cell-free tubes containing medium harvested from aerobically cultivated mammalian cells. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Increases in treponemal numbers were observed using dark-field microscopy but were not substantiated using the rabbit lesion test. Continuous passage of the treponeme was not achieved in vitro.


Author(s):  
Lidia Stasiak-Różańska ◽  
Anna Berthold-Pluta ◽  
Antoni Stanisław Pluta ◽  
Krzysztof Dasiewicz ◽  
Monika Garbowska

Probiotics are recommended, among others, in the diet of children who are under antibiotic therapy, or that suffer from food allergies or travel diarrhea, etc. In the case of toddlers taking probiotic preparations, it is highly recommended to first remove the special capsule, which normally protects probiotic strains against hard conditions in the gastrointestinal tract. Otherwise, the toddler may choke. This removal can impair probiotic survival and reduce its efficacy in a toddler’s organism. The aim of this study was to evaluate the survivability of five strains of lactic acid bacteria from the commercial probiotics available on the Polish market under simulated conditions of the gastrointestinal tract. Five probiotics (each including one of these strains: Bifidobacterium BB-12, Lactobacillus (Lb.) rhamnosus GG, Lb. casei, Lb. acidophilus, Lb. plantarum) were protective capsule deprived, added in a food matrix (chicken–vegetable soup) and subjected under simulated conditions of the gastric and gastrointestinal passage. Strain survivability and possibility to growth were evaluated. Obtained results showed that, among all analyzed commercial probiotic strains, the Lb. plantarum was the most resistant to the applied conditions of the culture medium. They showed a noticeable growth under both in vitro gastric conditions at pH 4.0 and 5.0, as well as in vitro intestinal conditions at all tested concentrations of bile salts.


1991 ◽  
Vol 125 (3) ◽  
pp. 280-285 ◽  
Author(s):  
J. Alan Talbot ◽  
Ann Lambert ◽  
Robert Mitchell ◽  
Marek Grabinski ◽  
David C. Anderson ◽  
...  

Abstract We have investigated the role of Ca2+ in the control of FSH-induced estradiol secretion by Sertoli cells isolated from 8-10 days old rats. Exogenous Ca2+ (4-8 mmol/1) inhibited FSH-stimulated E2 secretion such that, with 8 mmol/l Ca2+ and FSH (8 IU/l) E2 secretion decreased from 2091±322 to 1480±84 pmol/l (p<0.002), whilst chelation of Ca2+ in the culture medium with EGTA (3 mmol/l) increased E2 secretion from 360±45 to 1242±133 pmol/l) in the absence of FSH. Further, EGTA (3 mmol/l) markedly potentiated FSH (8 IU/l), forskolin (1 μmol/l) and dibutyryl cAMP (1 mmol/l)-stimulated E2 secretion. Addition of the Ca2+ ionophores, ionomycin (2-5 μmol/l) and A23187 (2 μmol/l), inhibited FSH (8 IU/l)-stimulated E2 secretion by >80%. The effect of ionomycin was totally reversible, whereas that of A23187 was irreversible. Ionomycin (5 μmol/l) had no effect on EGTA-induced E2 secretion in the absence of FSH, but reduced EGTA-provoked E2 secretion by 59% in the presence of FSH (8 IU/l). Similarly, forskolin- and dibutyryl cAMP-provoked E2 production was inhibited 46-50% by ionomycin (5 μmol/l). We conclude that FSH-induced E2 secretion from immature rat Sertoli cells is modulated by intra- and extracellular Ca2+.


Reproduction ◽  
2000 ◽  
pp. 391-396 ◽  
Author(s):  
AH Duittoz ◽  
M Batailler

The aim of this study was to investigate the development of pulsatile GnRH secretion by GnRH neurones in primary cultures of olfactory placodes from ovine embryos. Culture medium was collected every 10 min for 8 h to detect pulsatile secretion. In the first experiment, pulsatile secretion was studied in two different sets of cultures after 17 and 24 days in vitro. In the second experiment, a set of cultures was tested after 10, 17 and 24 days in vitro to investigate the development of pulsatile GnRH secretion in each individual culture. This study demonstrated that (i) primary cultures of GnRH neurones from olfactory explants secreted GnRH in a pulsatile manner and that the frequency and mean interpulse duration were similar to those reported in castrated ewes, and (ii) pulsatile secretion was not present at the beginning of the culture but was observed between 17 and 24 days in vitro, indicating the maturation of individual neurones and the development of their synchronization.


Author(s):  
С.В. Калиш ◽  
С.В. Лямина ◽  
А.А. Раецкая ◽  
И.Ю. Малышев

Цель исследования. Репрограммирование М1 фенотипа макрофагов с ингибированными факторами транскрипции М2 фенотипа STAT3, STAТ6 и SMAD и оценка их влияния на развитие карциномы Эрлиха (КЭ) in vitro и in vivo. Методика. Рост опухоли иницировали in vitro путем добавления клеток КЭ в среду культивирования RPMI-1640 и in vivo путем внутрибрюшинной инъекции клеток КЭ мышам. Результаты. Установлено, что M1макрофаги и in vitro, и in vivo оказывают выраженный противоопухолевый эффект, который превосходит антиопухолевые эффекты М1, M1, M1 макрофагов и цисплатина. Заключение. М1 макрофаги с ингибированными STAT3, STAT6 и/или SMAD3 эффективно ограничивают рост опухоли. Полученные данные обосновывают разработку новой технологии противоопухолевой клеточной терапии. Objective. Reprogramming of M1 macrophage phenotype with inhibited M2 phenotype transcription factors, such as STAT3, STAT6 and SMAD and assess their impact on the development of Ehrlich carcinoma (EC) in vitro and in vivo . Methods. Tumor growth in vitro was initiated by addition of EC cells in RPMI-1640 culture medium and in vivo by intraperitoneal of EC cell injection into mice. Results. It was found that M1 macrophages have a pronounced anti-tumor effect in vitro , and in vivo , which was greater than anti-tumor effects of M1, M1, M1 macrophages and cisplatin. Conclusion. M1 macrophages with inhibited STAT3, STAT6 and/or SMAD3 effectively restrict tumor growth. The findings justify the development of new anti-tumor cell therapy technology.


Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.


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