In Vitro Metabolic Depression of Tissues from the Aestivating Frog Neobatrachus Pelobatoides

1991 ◽  
Vol 161 (1) ◽  
pp. 273-283 ◽  
Author(s):  
JAMES E. FLANIGAN ◽  
PHILIP C. WITHERS ◽  
MICHAEL GUPPY
Keyword(s):  
Energy Demand ◽  
Energy Production ◽  
Metabolic Depression ◽  
Short Term ◽  
Vitro Assay ◽  
Summer Dormancy ◽  
Resting Metabolism ◽  
Assay Conditions

The desert frog Neobatrachus pelobatoides reduced its resting metabolism in vivo by 60–70% during 5–7 weeks of aestivation (summer dormancy). The rate ofoxygen consumption (V·OO2) of isolated and intact skeletal muscle, measured in vitro, was 70% lower for aestivating frogs compared with non-aestivating frogs. The cause of the reduced V·OO2 of aestivating frog muscle must lie in the tissue itself rather than being induced by external factors such as oxygen supply or bloodborne metabolites (because these were identical in the in vitro assay conditions), by any short-term effects produced by hormones (as these would have been washed out of the tissues during incubation) or by tissue dehydration (as the tissues from aestivating frogs had rehydrated to non-aestivating levels). The reduced in vitro muscle V·OO2 accounted for 60–77% of the frogs in vivo metabolic depression that accompanied aestivation. Other tissues of the aestivating frog, namely intestine, liver, skin and fat, did not have a reduced in vitro V·OO2. We suggest that metabolic depression is initiated by reduced energy demand in cells and this consequently leads to reduced energy production.

Glucuronidation of thyroid hormone in rat liver: effects of in vivo treatment with microsomal enzyme inducers and in vitro assay conditions.

Endocrinology ◽  
1993 ◽  
Vol 133 (5) ◽  
pp. 2177-2186 ◽  
Author(s):  
T J Visser ◽  
E Kaptein ◽  
H van Toor ◽  
J A van Raaij ◽  
K J van den Berg ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Rat Liver ◽  
In Vitro Assay ◽  
Microsomal Enzyme ◽  
Vitro Assay ◽  
In Vivo Treatment ◽  
Microsomal Enzyme Inducers ◽  
Assay Conditions

Case study in 21 st century ecotoxicology: using in vitro aromatase inhibition data to predict short term in vivo responses in adult female fish

2020 ◽  
Author(s):  
Daniel L. Villeneuve ◽  
Brett R. Blackwell ◽  
Jenna E. Cavallin ◽  
Wan‐Yun Cheng ◽  
David J. Feifarek ◽  
...  
Keyword(s):  
Adult Female ◽  
Female Fish ◽  
Aromatase Inhibition ◽  
Short Term ◽  
Inhibition Data

scholarly journals TNF-α Triggers RIP1/FADD/Caspase-8-Mediated Apoptosis of Astrocytes and RIP3/MLKL-Mediated Necroptosis of Neurons Induced by Angiostrongylus cantonensis Infection

2021 ◽  
Author(s):  
Hongli Zhou ◽  
Minyu Zhou ◽  
Yue Hu ◽  
Yanin Limpanon ◽  
Yubin Ma ◽  
...  
Keyword(s):  
Cell Death ◽  
Enrichment Analysis ◽  
Angiostrongylus Cantonensis ◽  
Gene Set Enrichment Analysis ◽  
Caspase 8 ◽  
Cns Injury ◽  
Vitro Assay ◽  
Tnf Α

AbstractAngiostrongylus cantonensis (AC) can cause severe eosinophilic meningitis or encephalitis in non-permissive hosts accompanied by apoptosis and necroptosis of brain cells. However, the explicit underlying molecular basis of apoptosis and necroptosis upon AC infection has not yet been elucidated. To determine the specific pathways of apoptosis and necroptosis upon AC infection, gene set enrichment analysis (GSEA) and protein–protein interaction (PPI) analysis for gene expression microarray (accession number: GSE159486) of mouse brain infected by AC revealed that TNF-α likely played a central role in the apoptosis and necroptosis in the context of AC infection, which was further confirmed via an in vivo rescue assay after treating with TNF-α inhibitor. The signalling axes involved in apoptosis and necroptosis were investigated via immunoprecipitation and immunoblotting. Immunofluorescence was used to identify the specific cells that underwent apoptosis or necroptosis. The results showed that TNF-α induced apoptosis of astrocytes through the RIP1/FADD/Caspase-8 axis and induced necroptosis of neurons by the RIP3/MLKL signalling pathway. In addition, in vitro assay revealed that TNF-α secretion by microglia increased upon LSA stimulation and caused necroptosis of neurons. The present study provided the first evidence that TNF-α was secreted by microglia stimulated by AC infection, which caused cell death via parallel pathways of astrocyte apoptosis (mediated by the RIP1/FADD/caspase-8 axis) and neuron necroptosis (driven by the RIP3/MLKL complex). Our research comprehensively elucidated the mechanism of cell death after AC infection and provided new insight into targeting TNF-α signalling as a therapeutic strategy for CNS injury.

scholarly journals Isolation in a single step of a highly enriched murine hematopoietic stem cell population with competitive long-term repopulating ability

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 930-939 ◽  
Author(s):  
SJ Szilvassy ◽  
PM Lansdorp ◽  
RK Humphries ◽  
AC Eaves ◽  
CJ Eaves
Keyword(s):  
Simple Procedure ◽  
Hematopoietic Cells ◽  
Stem Cell Population ◽  
Proliferative Potential ◽  
Vitro Assay ◽  
Hematopoietic Stem ◽  
Marrow Cells

Abstract A simple procedure is described for the quantitation and enrichment of murine hematopoietic cells with the capacity for long-term repopulation of lymphoid and myeloid tissues in lethally irradiated mice. To ensure detection of the most primitive marrow cells with this potential, we used a competitive assay in which female recipients were injected with male “test” cells and 1 to 2 x 10(5) “compromised” female marrow cells with normal short-term repopulating ability, but whose long-term repopulating ability had been reduced by serial transplantation. Primitive hematopoietic cells were purified by flow cytometry and sorting based on their forward and orthogonal light-scattering properties, and Thy-1 and H-2K antigen expression. Enrichment profiles for normal marrow, and marrow of mice injected with 5-fluorouracil (5- FU) four days previously, were established for each of these parameters using an in vitro assay for high proliferative potential, pluripotent colony-forming cells. When all four parameters were gated simultaneously, these clonogenic cells were enriched 100-fold. Both day 9 and day 12 CFU-S were copurified; however, the purity (23%) and enrichment (75-fold) of day 12 CFU-S in the sorted population was greater with 5-FU-treated cells. Five hundred of the sorted 5-FU marrow cells consistently repopulated recipient lymphoid and myeloid tissues (greater than 50% male, 1 to 3 months post-transplant) when co-injected with 1 to 2 x 10(5) compromised female marrow cells, and approximately 100 were sufficient to achieve the same result in 50% of recipients under the same conditions. This relatively simple purification and assay strategy should facilitate further analysis of the heterogeneity and regulation of stem cells that maintain hematopoiesis in vivo.

scholarly journals DIGESTIBLE ENERGY IN FORAGE BYIN VIVOANDIN VITROASSAYS

1970 ◽  
Vol 50 (3) ◽  
pp. 557-562 ◽  
Author(s):  
J. E. TROELSEN
Keyword(s):  
Energy Analysis ◽  
Energy Content ◽  
In Vitro Assay ◽  
Pure Species ◽  
Vitro Assay ◽  
Digestible Energy ◽  
The Relationship ◽  
Cattle And Sheep

Forage of six pure species was harvested for hay at several maturity stages during four years. The digestible energy content of 102 different lots of hay was determined by feeding to four groups of sheep during the same period, and by in vitro digestions and energy analysis of the undigested residues. The relationship between digestible energy content assayed by the two methods was highly significant (r = 0.85) and did not differ between years and species. Exclusion from regression of the hays containing less than 2 or more than 3 digestible kcal/g revealed that the in vitro assay could reproduce the in vivo digestible energy value with a standard deviation of 0.31 in over 70% of the hays. This represented the maturity and quality range of forage commonly fed to cattle and sheep. The in vitro assay therefore appeared promising for commercial quality determinations.

scholarly journals SBA1 Encodes a Yeast Hsp90 Cochaperone That Is Homologous to Vertebrate p23 Proteins

1998 ◽  
Vol 18 (7) ◽  
pp. 3727-3734 ◽  
Author(s):  
Yifang Fang ◽  
Albert E. Fliss ◽  
Jie Rao ◽  
Avrom J. Caplan
Keyword(s):  
Hormone Receptors ◽  
Pcr Amplification ◽  
Steroid Hormone Receptors ◽  
Loss Of Function ◽  
Vitro Assay ◽  
Growth Defects ◽  
Double Deletion ◽  
Affinity Isolation

ABSTRACT The Saccharomyces cerevisiae SBA1 gene was cloned by PCR amplification from yeast genomic DNA following its identification as encoding an ortholog of human p23, an Hsp90 cochaperone. TheSBA1 gene product is constitutively expressed and nonessential, although a disruption mutant grew more slowly than the wild type at both 18 and 37°C. A double deletion of SBA1and STI1, encoding an Hsp90 cochaperone, displayed synthetic growth defects. Affinity isolation of histidine-tagged Sba1p (Sba1His6) after expression in yeast led to coisolation of Hsp90 and the cyclophilin homolog Cpr6. Using an in vitro assembly assay, purified Sba1His6 bound to Hsp90 only in the presence of adenosine 5′-O-(3-thiotriphosphate) or adenyl-imidodiphosphate. Furthermore, interaction between purified Sba1His6 and Hsp90 in yeast extracts was inhibited by the benzoquinoid ansamycins geldanamycin and macbecin. The in vitro assay was also used to identify residues in Hsp90 that are important for complex formation with Sba1His6, and residues in both the N-terminal nucleotide binding domain and C-terminal half were characterized. In vivo analysis of known Hsp90 substrate proteins revealed that Sba1 loss of function had only a mild effect on the activity of the tyrosine kinase v-Src and steroid hormone receptors.

PREDICTION OF DIGESTIBLE ENERGY INTAKE BY SHEEP OF DIETS CONTAINING GROUND ROUGHAGE AND GRAIN

1977 ◽  
Vol 57 (2) ◽  
pp. 279-288 ◽  
Author(s):  
S. O. THORLACIUS
Keyword(s):  
Energy Demand ◽  
Dry Matter ◽  
Organic Matter Content ◽  
Rapeseed Meal ◽  
Regression Equation ◽  
Curvilinear Relationship ◽  
Crested Wheatgrass ◽  
Digestible Energy

Digestibility and intake of diets containing 8, 28, 48 or 68% ground wheat straw plus ground crested wheatgrass and rapeseed meal, and diets containing 33, 48, 63 and 78% ground crested wheatgrass plus barley and rapeseed meal was measured with four yearling wethers per diet. Digestible energy (DE) content ranged from 2.07 to 2.95 kcal/g dry matter (DM) and dry matter digestibility (DMD) (%) from 48.7 to 71.1%. Regression of DE intake y (kcal/w0.75kg/d) on DE content (x) was curvilinear; y = −2,133 + 1,626x − 277.9x2, r = 0.996, P < 0.01, SE = ± 7.3. There was also a curvilinear relationship between diet density, as fed, (x) g (DM)/ml and DMD (%), y = 9.057 + 364.1x − 530.0x2, r = 0.970, P < 0.01, SE = ± 2.4. A linear regression equation was calculated over the DE range (2.07–2.52) for which there was an obvious increase in DE intake with increasing diet DE content; y = −700.6 + 361x, r = 0.994, P < 0.01, SE = ± 9.4, y = DE intake (kcal/w0.75kg/d), x = DE [kcal/g (DM)]. Using this regression equation and assuming a linear increase in DE intake with increase in diet DE content up to a point at which the apparent energy demand of the animal is satisfied gave a more accurate prediction of DE intake than when the curvilinear regression equation, y = −2,133 + 1,626x − 277.9x2, was used empirically. Accuracy of the prediction was further improved by expressing DE/unit ration volume instead of per unit DM. The sheep used in the present experiments had an apparent energy demand of 230 kcal/w0.75kg/day which was met at diet DE contents above 0.48 kcal/ml or 2.6 kcal/g (DM). There was a high correlation between in vivo DE content of the diet, y [kcal/g (DM)] and in vitro (x) digestible organic matter content, x, (%), r = 0.991, P < 0.01, y = 0.38 + 0.037x, SE = ± 0.04.

scholarly journals Mutagenicity evaluation of Anastatica hierochuntica L. aqueous extract in vitro and in vivo

2017 ◽  
Vol 243 (4) ◽  
pp. 375-385 ◽  
Author(s):  
Siti Rosmani Md Zin ◽  
Zahurin Mohamed ◽  
Mohammed A Alshawsh ◽  
Won F Wong ◽  
Normadiah M Kassim
Keyword(s):  
Aqueous Extract ◽  
Positive Control ◽  
In Vitro Assay ◽  
In Vivo Study ◽  
Sufficient Evidence ◽  
Aqueous Extracts ◽  
Mutagenic Potential ◽  
Vitro Assay

Anastatica hierochuntica L. ( A. hierochuntica), a folk medicinal plant, was evaluated for mutagenic potential via in vitro and in vivo assays. The in vitro assay was conducted according to modified Ames test, while the in vivo study was performed according to Organisation for Economic Co-operation and Development guideline for mammalian erythrocyte micronucleus assay. Four groups ( n= 5 males and 5 females per group) Sprague Dawley rats were randomly chosen as the negative control, positive control (received a single intramuscular injection of cyclophosphamide 50 mg/kg), 1000 and, 2000 mg/kg A. hierochuntica aqueous extracts. All groups except the positive control were treated orally for three days. Findings of the in vitro assay showed mutagenic potential of AHAE at 0.04 and 0.2 mg/ml. However, no mutagenic effect was demonstrated in the in vivo study up to 2000 mg/kg. No significant reduction in the polychromatic and normochromatic erythrocytes ratio was noted in any of the groups. Meanwhile, high micronucleated polychromatic erythrocytes frequency was seen in cyclophosphamide-treated group only. These findings could perhaps be due to insufficient dosage of A. hierochuntica aqueous extracts to cause genetic damage on the bone marrow target cells. Further acute and chronic in vivo toxicity studies may be required to draw pertinent conclusion on the safety aspect of A. hierochuntica aqueous extracts consumption. Impact statement In this paper, we report on the mutagenicity evaluation of Anastatica hierochuntica aqueous extract. This is a significant research in view of the popularity of this herb consumption by the people across the globe despite of limited scientific evidence on its toxicity potential. This study is intended to encourage more extensive related research in order to provide sufficient evidence and guidance for determining its safe dosage.

A NEW NON-PROTEIN NITROGEN SOURCE FOR RUMINANTS

1969 ◽  
Vol 49 (2) ◽  
pp. 135-141 ◽  
Author(s):  
L. P. Milligan ◽  
A. R. Robblee ◽  
J. C. Wood ◽  
W. C. Kay ◽  
S. K. Chakrabartty
Keyword(s):  
Nitrogen Source ◽  
Dietary Supplement ◽  
Nitrogen Retention ◽  
Feeding Trial ◽  
Short Term ◽  
Protein Nitrogen ◽  
Rumen Microorganisms ◽  
Production Of Ammonia

The preparation of a polymer of urea and furfural containing 23.2% nitrogen is described. This product was converted by rumen microorganisms in vitro to ammonia at a rate approximately one-seventh that of conversion of urea to ammonia. Use of the polymer as a dietary supplement in a feeding trial with lambs improved nitrogen retention over that of unsupplemented controls by 3.45 g of nitrogen retained per day, while an isonitrogenous quantity of supplemental urea improved nitrogen retention by 0.51 g of nitrogen retained per day. The blood urea pattern, throughout the day, of lambs adapted to control, urea-supplemented and urea–furfural polymer-supplemented rations indicated a slow, prolonged production of ammonia from the latter supplement and very rapid, short-term degradation of urea in vivo.

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