scholarly journals A Multiparametric Assessment of Human Islets Predicts Transplant Outcomes in Diabetic Mice

2021 ◽  
Vol 30 ◽  
pp. 096368972110522
Author(s):  
Hirotake Komatsu ◽  
Meirigeng Qi ◽  
Nelson Gonzalez ◽  
Mayra Salgado ◽  
Leonard Medrano ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
Single Cells ◽  
Human Islets ◽  
In Vitro Assays ◽  
Diabetic Mice ◽  
Post Transplantation

Prior to transplantation into individuals with type 1 diabetes, in vitro assays are used to evaluate the quality, function and survival of isolated human islets. In addition to the assessments of these parameters in islet, they can be evaluated by multiparametric morphological scoring (0–10 points) and grading (A, B, C, D, and F) based on islet characteristics (shape, border, integrity, single cells, and diameter). However, correlation between the multiparametric assessment and transplantation outcome has not been fully elucidated. In this study, 55 human islet isolations were scored using this multiparametric assessment. The results were correlated with outcomes after transplantation into immunodeficient diabetic mice. In addition, the multiparametric assessment was compared with oxygen consumption rate of isolated islets as a potential prediction factor for successful transplantations. All islet batches were assessed and found to score: 9 points ( n = 18, Grade A), 8 points ( n = 19, Grade B), and 7 points ( n = 18, Grade B). Islets that scored 9 (Grade A), scored 8 (Grade B) and scored 7 (Grade B) were transplanted into NOD/SCID mice and reversed diabetes in 81.2%, 59.4%, and 33.3% of animals, respectively ( P < 0.0001). Islet scoring and grading correlated well with glycemic control post-transplantation ( P < 0.0001) and reversal rate of diabetes ( P < 0.05). Notably, islet scoring and grading showed stronger correlation with transplantation outcome compared to oxygen consumption rate. Taken together, a multiparametric assessment of isolated human islets was highly predictive of transplantation outcome in diabetic mice.

43 EFFECT OF VITRIFICATION AT GERMINAL VESICLE STAGE ON THE MITOCHONDRIAL AND CYTOSKELETAL INTEGRITY IN BOVINE OOCYTES

2012 ◽  
Vol 24 (1) ◽  
pp. 134
Author(s):  
Y. Abe ◽  
K. Takakura ◽  
K. Kaito ◽  
T. Ogawa ◽  
M. Yokoo ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
Germinal Vesicle ◽  
Nylon Mesh ◽  
Developmental Rates ◽  
Type 3

In the previous report, we demonstrated that bovine germinal vesicle (GV) stage oocytes vitrified using a nylon mesh holder showed an in vitro maturation rate to the metaphase II (MII) stage similar to that of fresh ones. However, cleavage and developmental rates of vitrified oocytes were low. Because mitochondria and the cytoskeleton are thought to have a central role in energy supply and cellular division in mammalian embryogenesis, it seems possible that alternation in their function in vitrified GV oocytes may contribute to subsequent lower cleavage and developmental rates. The oxygen consumption rate reflects the mitochondrial activity and its measurement may be an effective way for non-invasive evaluation of oocyte quality. In this study, to ascertain that altered mitochondrial functions and cytoskeleton may contribute to reduce the quality of oocytes after vitrification, we evaluated the distribution of active mitochondria and the cytoskeleton in vitrified oocytes. We also examined the relationship between oxygen consumption rate and the distribution of active mitochondria in vitrified oocytes. Bovine GV oocytes connected with cumulus cells were exposed to the cryoprotectant (EFS40) in a stepwise way and transferred onto a nylon mesh holder, followed by plunging them directly into liquid nitrogen. After warming, vitrified oocytes were allowed in vitro maturation. After denuding, matured oocytes were stained with a mitochondria-specific probe, rhodamine-123 and then oxygen consumption rate using an embryo respirometer (HV-403; Research Institute for Functional Peptides, Yamagata, Japan) was measured in each oocyte. According to morphological distribution of mitochondria, oocytes were classified as follows: type 1, uniform distribution; type 2, spotted distribution; and type 3, a weak fluorescence. The oxygen consumption rate of the fresh oocytes at the MII stage was significantly (P < 0.05) higher than that of vitrified oocytes (5.24 and 4 × 1015 mol–1 s–1, respectively), although there was no difference between the fresh and vitrified groups at the GV stage (5.02 and 5.06 × 1015 mol–1 s–1, respectively). The oxygen consumption rates of type 1 oocytes in fresh and vitrified groups at the MII stage tended to be higher than those of type 2 and 3 oocytes (type 1, 5.29 and 5.27; type 2, 4.99 and 4.52; type 3, 4.77 and 4.48 × 1015 mol–1 s–1, respectively). In addition, the percentage of type 1 oocytes in the fresh group was significantly (P < 0.05) higher than that in the vitrified group (59.4 and 34.3%, respectively). The matured oocytes also were stained with α-tubulin monoclonal antibody or F-phalloidin independently to examine the morphological status of microtubules or microfilament. The rates of oocytes with abnormal microtubules and microfilament in the vitrified group were 29.7 and 43.5%, respectively, showing higher rates compared with corresponding fresh oocytes (9.8%; P < 0.05 and 25.0%; P = 0.21, respectively). These results suggested that the reduction of quality and subsequent developmental competence in vitrified oocytes might be related to damages of mitochondria and cytoskeleton.

scholarly journals A Peripheral CB1R Antagonist Increases Lipolysis, Oxygen Consumption Rate, and Markers of Beiging in 3T3-L1 Adipocytes Similar to RIM, Suggesting that Central Effects Can Be Avoided

2020 ◽  
Vol 21 (18) ◽  
pp. 6639
Author(s):  
Rebecca L. Paszkiewicz ◽  
Richard N. Bergman ◽  
Roberta S. Santos ◽  
Aaron P. Frank ◽  
Orison O. Woolcott ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Oxygen Consumption ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
Cannabinoid Receptor ◽  
In Vitro Assays ◽  
Mitochondrial Uncoupling ◽  
Direct Role ◽  
The Impact

With the increased prevalence of obesity and related co-morbidities, such as type 2 diabetes (T2D), worldwide, improvements in pharmacological treatments are necessary. The brain- and peripheral-cannabinoid receptor 1 (CB1R) antagonist rimonabant (RIM) has been shown to induce weight loss and improve glucose homeostasis. We have previously demonstrated that RIM promotes adipose tissue beiging and decreased adipocyte cell size, even during maintenance on a high-fat diet. Given the adverse side-effects of brain-penetrance with RIM, in this study we aimed to determine the site of action for a non-brain-penetrating CB1R antagonist AM6545. By using in vitro assays, we demonstrated the direct effects of this non-brain-penetrating CB1R antagonist on cultured adipocytes. Specifically, we showed, for the first time, that AM6545 significantly increases markers of adipose tissue beiging, mitochondrial biogenesis, and lipolysis in 3T3-L1 adipocytes. In addition, the oxygen consumption rate (OCR), consisting of baseline respiratory rate, proton leak, maximal respiratory capacity, and ATP synthase activity, was greater for cells exposed to AM6545, demonstrating greater mitochondrial uncoupling. Using a lipolysis inhibitor during real-time OCR measurements, we determined that the impact of CB1R antagonism on adipocytes is driven by increased lipolysis. Thus, our data suggest the direct role of CB1R antagonism on adipocytes does not require brain penetrance, supporting the importance of focus on peripheral CB1R antagonism pharmacology for reducing the incidence of obesity and T2D.

scholarly journals Necrostatin-1 Supplementation to Islet Tissue Culture Enhances the In-Vitro Development and Graft Function of Young Porcine Islets

2021 ◽  
Vol 22 (16) ◽  
pp. 8367
Author(s):  
Hien Lau ◽  
Shiri Li ◽  
Nicole Corrales ◽  
Samuel Rodriguez ◽  
Mohammadreza Mohammadi ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Oxygen Consumption ◽  
Insulin Secretion ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
In Vitro Development ◽  
Porcine Islets ◽  
Vitro Development

Pre-weaned porcine islets (PPIs) represent an unlimited source for islet transplantation but are functionally immature. We previously showed that necrostatin-1 (Nec-1) immediately after islet isolation enhanced the in vitro development of PPIs. Here, we examined the impact of Nec-1 on the in vivo function of PPIs after transplantation in diabetic mice. PPIs were isolated from pancreata of 8–15-day-old, pre-weaned pigs and cultured in media alone, or supplemented with Nec-1 (100 µM) on day 0 or on day 3 of culture (n = 5 for each group). On day 7, islet recovery, viability, oxygen consumption rate, insulin content, cellular composition, insulin secretion capacity, and transplant outcomes were evaluated. While islet viability and oxygen consumption rate remained high throughout 7-day tissue culture, Nec-1 supplementation on day 3 significantly improved islet recovery, insulin content, endocrine composition, GLUT2 expression, differentiation potential, proliferation capacity of endocrine cells, and insulin secretion. Adding Nec-1 on day 3 of tissue culture enhanced the islet recovery, proportion of delta cells, beta-cell differentiation and proliferation, and stimulation index. In vivo, this leads to shorter times to normoglycemia, better glycemic control, and higher circulating insulin. Our findings identify the novel time-dependent effects of Nec-1 supplementation on porcine islet quantity and quality prior to transplantation.

THE IN VITRO ACTION OF ACTH ON THE OXYGEN CONSUMPTION OF SLICES OF CATTLE ADRENAL CORTEX

1953 ◽  
Vol 9 (4) ◽  
pp. 379-390 ◽  
Author(s):  
M. REISS ◽  
EVA BRUMMEL ◽  
I. D. K. HALKERSTON ◽  
F. E. BADRICK ◽  
M. FENWICK
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Uptake ◽  
Linear Relationship ◽  
Adrenal Cortex ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
Biological Assay ◽  
Percentage Increase ◽  
Small Doses

A technique for measuring the action of small doses of ACTH on the oxygen consumption of slices of cattle adrenal cortex is described. The oxygen consumption rate of such slices in vitro is increased by ACTH. A linear relationship between logarithm of the dose of ACTH and the percentage increase in the rate of oxygen uptake is obtained with this method, and its suitability for biological assay purposes has been investigated. The question of the specificity of this action of ACTH is discussed.

Development of a new oxygen consumption rate assay in cultures of Acanthamoeba (Protozoa: Lobosea) and its application to evaluate viability and amoebicidal activity in vitro

2015 ◽  
Vol 155 ◽  
pp. 35-39 ◽  
Author(s):  
I. Heredero-Bermejo ◽  
A. Criado-Fornelio ◽  
J. Soliveri ◽  
J.A. Díaz-Martín ◽  
J. Matilla-Fuentes ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
Amoebicidal Activity

scholarly journals Event-Associated Oxygen Consumption Rate Increases ca. Five-Fold When Interictal Activity Transforms into Seizure-Like Events In Vitro

2017 ◽  
Vol 18 (9) ◽  
pp. 1925 ◽  
Author(s):  
Karl Schoknecht ◽  
Nikolaus Berndt ◽  
Jörg Rösner ◽  
Uwe Heinemann ◽  
Jens Dreier ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Consumption Rate ◽  
Consumption Rate

Oxygen consumption by bovine granulosa cells with prediction of oxygen transport in preantral follicles

2013 ◽  
Vol 25 (8) ◽  
pp. 1158 ◽  
Author(s):  
Dongxing Li ◽  
Gabe P. Redding ◽  
John E. Bronlund
Keyword(s):  
Oxygen Consumption ◽  
Granulosa Cells ◽  
Oxygen Transport ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
Model Parameters ◽  
Preantral Follicles ◽  
Rate Of Oxygen Consumption ◽  
Key Parameter

The rate of oxygen consumption by granulosa cells is a key parameter in mathematical models that describe oxygen transport across ovarian follicles. This work measured the oxygen consumption rate of bovine granulosa cells in vitro to be in the range 2.1–3.3 × 10–16 mol cell–1 s–1 (0.16–0.25 mol m–3 s–1). The implications of the rates for oxygen transport in large bovine preantral follicles were examined using a mathematical model. The results indicate that oocyte oxygenation becomes increasingly constrained as preantral follicles grow, reaching hypoxic levels near the point of antrum formation. Beyond a preantral follicle radius of 134 µm, oxygen cannot reach the oocyte surface at typical values of model parameters. Since reported sizes of large bovine preantral follicles range from 58 to 145 µm in radius, this suggests that oocyte oxygenation is possible in all but the largest preantral follicles, which are on the verge of antrum formation. In preantral bovine follicles, the oxygen consumption rate of granulosa cells and fluid voidage will be the key determinants of oxygen levels across the follicle.

scholarly journals Ceria nanoparticles ameliorate renal fibrosis by modulating the balance between oxidative phosphorylation and aerobic glycolysis

2022 ◽  
Vol 20 (1) ◽  
Author(s):  
Mengling Wang ◽  
Feng Zeng ◽  
Fengling Ning ◽  
Yinhang Wang ◽  
Shilin Zhou ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Consumption Rate ◽  
Renal Fibrosis ◽  
Consumption Rate ◽  
Metabolic Reprogramming ◽  
Ceria Nanoparticles ◽  
Extracellular Acidification ◽  
Extracellular Acidification Rate

Abstract Background and aims Renal fibrosis is the common outcome in all progressive forms of chronic kidney disease. Unfortunately, the pathogenesis of renal fibrosis remains largely unexplored, among which metabolic reprogramming plays an extremely crucial role in the evolution of renal fibrosis. Ceria nanoparticles (CeNP-PEG) with strong ROS scavenging and anti-inflammatory activities have been applied for mitochondrial oxidative stress and inflammatory diseases. The present study aims to determine whether CeNP-PEG has therapeutic value for renal fibrosis. Methods The unilateral ureteral obstructive fibrosis model was used to assess the therapeutic effects in vivo. Transforming growth factor beta1-induced epithelial-to-mesenchymal transition in HK-2 cells was used as the in vitro cell model. The seahorse bioscience X96 extracellular flux analyzer was used to measure the oxygen consumption rate and extracellular acidification rate. Results In the present study, CeNP-PEG treatment significantly ameliorated renal fibrosis by increased E-cadherin protein expression, and decreased α-SMA, Vimentin and Fibronectin expression both in vitro and in vivo. Additionally, CeNP-PEG significantly reduced the ROS formation and improved the levels of mitochondrial ATP. The seahorse analyzer assay demonstrated that the extracellular acidification rate markedly decreased, whereas the oxygen consumption rate markedly increased, in the presence of CeNP-PEG. Furthermore, the mitochondrial membrane potential markedly enhanced, hexokinase 1 and hexokinase 2 expression significantly decreased after treatment with CeNP-PEG. Conclusions CeNP-PEG can block the dysregulated metabolic status and exert protective function on renal fibrosis. This may provide another therapeutic option for renal fibrosis. Graphical Abstract

scholarly journals In Vitro Evaluation of the Combination of Melaleuca alternifolia (Tea Tree) Oil and Dimethyl Sulfoxide (DMSO) against Trophozoites and Cysts of Acanthamoeba Strains. Oxygen Consumption Rate (OCR) Assay as a Method for Drug Screening

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 491
Author(s):  
Tania Martín-Pérez ◽  
Irene Heredero-Bermejo ◽  
Cristina Verdú-Expósito ◽  
Jorge Pérez-Serrano
Keyword(s):  
Oxygen Consumption ◽  
Dimethyl Sulfoxide ◽  
Drug Screening ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
Screening Method ◽  
Acanthamoeba Keratitis ◽  
Tea Tree Oil ◽  
Tea Tree

Ameobae belonging to the genus Acanthamoeba are responsible for the human diseases Acanthamoeba keratitis (AK) and granulomatous amoebic encephalitis (GAE). The treatment of these illnesses is hampered by the existence of a resistance stage (cysts). In an attempt to add new agents that are effective against trophozoites and cysts, tea tree oil (TTO) and dimethyl sulfoxide (DMSO), separately and in combination, were tested In Vitro against two Acanthamoeba isolates, T3 and T4 genotypes. The oxygen consumption rate (OCR) assay was used as a drug screening method, which is to some extent useful in amoebicide drug screening; however, evaluation of lethal effects may be misleading when testing products that promote encystment. Trophozoite viability analysis showed that the effectiveness of the combination of both compounds is higher than when either compound is used alone. Therefore, the TTO alone or TTO + DMSO in combination were an amoebicide, but most of the amoebicidal activity in the combination’s treatments seemed to be caused mainly by the TTO effect. In fact, DMSO alone seems to be a non-amoebicide, triggering encystment. Regarding cytotoxicity, these compounds showed toxicity in human corneal epithelial cells (HCEpiC), even at low concentrations when tested in combination. In conclusion, the use of TTO and DMSO, in combination or alone, cannot be recommended as an alternative for AK treatment until more cytotoxicity and cyst adhesion tests are performed.

Sign in / Sign up

Export Citation Format

Share Document