Effects of parathyroid hormone on bone acid hydrolases in tissue culture

1968 ◽  
Vol 46 (2) ◽  
pp. 261-267 ◽  
Author(s):  
Susan Tolnai
Keyword(s):  
Parathyroid Hormone ◽  
Acid Phosphatase ◽  
Bone Cells ◽  
Culture Media ◽  
Enzyme Synthesis ◽  
Maximum Activity ◽  
Acid Protease ◽  
Acid Hydrolases ◽  
The Media

Experiments were carried out to assess the activity of acid hydrolases of bone cells under in vitro conditions. Rat and mouse skull bones were kept in vitro for 2 to 6 days in the presence and absence of added parathyroid hormone. The activities of acid protease (cathepsin D), acid phosphatase, and β-glucuronidase were measured in the culture media and in homogenates prepared from the cultured bones. All three enzymes showed increased activity in various degrees in the media when parathyroid hormone (PTE) was present, but increased enzyme synthesis in the homogenates could be detected only in the case of acid protease and acid phosphatase. The acid protease showed maximum activity at pH 3.0–3.2, and the PTE-stimulated increase was time-dependent. Lysozyme incorporated into the culture medium stimulated acid protease activity, while histone had an opposite effect. Cultures kept in incomplete media for a limited period of time also responded to PTE treatment with an increased release of acid hydrolases into the medium.

Biochemical markers for pregnancy in the spent culture medium of in vitro produced bovine embryos

2021 ◽  
Author(s):  
Gabriela de Oliveira Fernandes ◽  
Marcella Pecora Milazzotto ◽  
Andrei Antonioni Guedes Fidelis ◽  
Taynan Stonoga Kawamoto ◽  
Ligiane de Oliveira Leme ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Biochemical Markers ◽  
Culture Medium ◽  
Culture Media ◽  
Ionization Mass ◽  
Metabolic Profiles ◽  
Bovine Embryos ◽  
The Media

Abstract The present study aimed to identify biomarkers to assess the quality of in vitro produced (IVP) bovine embryos in the culture media. IVP embryos on Day (D) 5 of development were transferred to individual drops, where they were maintained for the last 48 h of culture. Thereafter, the medium was collected and the embryos were transferred to the recipients. After pregnancy diagnosis, the media were grouped into the pregnant and nonpregnant groups. The metabolic profiles of the media were analyzed via electrospray ionization mass spectrometry, and the concentrations of pyruvate, lactate, and glutamate were assessed using fluorimetry. The spectrometric profile revealed that the media from embryos from the pregnant group presented a higher signal intensity compared to that of the nonpregnant group; the ions 156.13 Da [M + H]+, 444.33 Da [M + H]+, and 305.97 Da [M + H]+ were identified as biomarkers. Spent culture medium from expanded blastocysts (Bx) that established pregnancy had a greater concentration of pyruvate (p = 0.0174) and lesser concentration of lactate (p = 0.042) than spent culture medium from Bx that did not establish pregnancy. Moreover, pyruvate in the culture media of Bx can predict pregnancy with 90.9% sensitivity and 75% specificity. In conclusion, we identified markers in the culture media that helped in assessing the most viable IVP embryos with a greater potential to establish pregnancy.

scholarly journals Gas Chromatography-Mass Spectrometry (GC-MS) Analysis of Essential Oils from AgNPs and AuNPs Elicited Lavandula angustifolia In Vitro Cultures

Molecules ◽  
2019 ◽  
Vol 24 (3) ◽  
pp. 606 ◽  
Author(s):  
Aneta Wesołowska ◽  
Paula Jadczak ◽  
Danuta Kulpa ◽  
Włodzimierz Przewodowski
Keyword(s):  
Molecular Weight ◽  
Silver Nanoparticles ◽  
Essential Oils ◽  
Culture Media ◽  
In Vitro Cultures ◽  
Gas Chromatography Mass Spectrometry ◽  
Lavandula Angustifolia ◽  
Gold And Silver Nanoparticles ◽  
The Media

The aim of this study was to determine how the addition of gold and silver nanoparticles to culture media affects the composition of essential oils extracted from Lavandula angustifolia propagated on MS media with the addition of 10 and 50 mg·dm−3 of gold (24.2 ± 2.4 nm) and silver (27.5 ± 4.8 nm) nanocolloids. The oil extracted from the lavender tissues propagated on the medium with 10 mg·dm−3 AgNPs (silver nanoparticles) differed the most with respect to the control; oil-10 compounds were not found at all, and 13 others were detected which were not present in the control oil. The addition of AuNPs (gold nanoparticles) and AgNPs to the media resulted in a decrease of lower molecular weight compounds (e.g., α- and β-pinene, camphene, δ-3-carene, p-cymene, 1,8-cineole, trans-pinocarveol, camphoriborneol), which were replaced by those of a higher molecular weight (τ- and α-cadinol 9-cedranone, cadalene, α-bisabolol, cis-14-nor-muurol-5-en-4-one, (E,E)-farnesol).

99 DIFFERENTIAL GENE REGULATION OF STEROIDOGENIC TRANSCRIPTS AND ESTRADIOL PRODUCTION FOLLOWING IN VITRO PIG EMBRYO ELONGATION IN ALGINATE HYDROGEL THREE-DIMENSIONAL MATRIX

2012 ◽  
Vol 24 (1) ◽  
pp. 162
Author(s):  
J. R. Miles ◽  
C. N. Sargus ◽  
S. A. Plautz ◽  
J. L. Vallet ◽  
A. K. Pannier
Keyword(s):  
Equal Opportunity ◽  
Culture Media ◽  
Morphological Changes ◽  
Three Dimensional ◽  
Differential Gene Regulation ◽  
Alginate Hydrogels ◽  
The Media ◽  
And Control

Between Day 10 and 12 of gestation, the pig embryo elongates from a sphere to a long thin, filament. During this time, the embryo increases the production of oestrogen via an increase in steroidogenic transcripts, which is critical for maternal recognition of pregnancy. To date, attempts to elongate porcine embryos in vitro have been unsuccessful. Therefore, the objective of this study was to utilise alginate hydrogels to establish a culture system that promotes in vitro embryo elongation with a corresponding increase in steroidogenic transcripts and oestradiol production. In 3 replicate collections, White crossbred gilts (n = 15) were bred at Day 0 of the oestrous cycle. At Day 9 of gestation, reproductive tracts were collected and flushed with RPMI-1640 containing antibiotics. Embryos were recovered, grouped according to size and washed with RPMI-1640 containing antibiotics and 10% fetal bovine serum (FBS). Embryos were randomly assigned to be encapsulated using a double encapsulation technique (0.7% sodium alginate and 1.5% calcium chloride solution) or used as controls. Encapsulated and control embryos were cultured for 96 h in CO2 -pretreated RPMI-1640 containing antibiotics and 10% FBS at 38°C, 5% CO2 in air and 100% humidity. Every 24 h, the embryos were imaged and half of the media was replaced. The removed media was stored at –20°C and used to assess oestradiol levels by radioimmunoassay. At the end of culture, a subset of encapsulated and control embryos were snap frozen and used to assess the expression level of steroidogenic transcripts (STAR, CYP11 and CYP19) using quantitative PCR. All data were analysed using general linear model (GLM) procedures for ANOVA. Cell survival, assessed by blastocyst fragmentation and confirmed by live/dead staining in representative embryos, was greater (P = 0.01) for encapsulated embryos (60.1 ± 4.8%) compared with controls (33.3 ± 4.8%). Of encapsulated embryos, 27% had some morphological change (minor flattening and tubal formation) and 14% had significant morphological changes (considerable flattening and tubal formation elongating through the gel), consistent with in vivo embryo elongation. In contrast, the control embryos had no morphological changes observed and remained spherical during culture. The expression levels of STAR, CYP11 and CYP19 were significantly (P < 0.05) greater in encapsulated embryos compared with control embryos. Furthermore, a significant (P < 0.01) time-dependent increase in oestradiol levels in the culture media of encapsulated embryos was identified compared with controls and culture media alone. These results illustrate that cultured pig embryos encapsulated in alginate hydrogels undergo limited morphological changes with increased expression of steroidogenic transcripts and oestrogen production. †USDA is an equal opportunity provider and employer.

Growth hormone and parathyroid hormone stimulate IGFBP-3 in rat osteoblasts

1994 ◽  
Vol 267 (2) ◽  
pp. E226-E233 ◽  
Author(s):  
C. Schmid ◽  
I. Schlapfer ◽  
M. Peter ◽  
M. Boni-Schnetzler ◽  
J. Schwander ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Parathyroid Hormone ◽  
Growth Factor ◽  
Culture Media ◽  
Insulin Like Growth Factor ◽  
Igf I ◽  
Conditioned Cell Culture Medium ◽  
Rat Osteoblasts ◽  
Igfbp 3

Osteoblast-like cells prepared from calvaria of newborn rats produce insulin-like growth factor (IGF) I and several insulin-like growth factor binding proteins (IGFBPs) in vitro. Among the IGFBPs found in conditioned cell culture medium, IGFBP-3 is the most abundant. Intact IGFBP-3, as assessed by 125I-labeled IGF-II ligand blot analysis, is more abundant in culture media of cells exposed to growth hormone (GH) or to parathyroid hormone (PTH), both at 5 x 10(-9) mol/l, for 24 h. At the same time, concentrations of IGF-I are increased in media of cells exposed to PTH but not to GH, compared with hormone-free control cultures. IGFBP-3 mRNA is increased in osteoblasts exposed to PTH or to GH but not in response to 5 x 10(-9) mol/l IGF-I. PTH exerts a rapid (within 2 h) stimulatory effect on IGF-I and IGFBP-3 production, both at the message and peptide levels, whereas GH increases only IGFBP-3, both at the message and peptide levels (after 24 h). We conclude that IGF-I does not mediate increased IGFBP-3 production by rat osteoblasts in response to GH and PTH.

scholarly journals Urochloa brizantha cv. Marandu presents a better response to in vitro salt stress than other commercial cultivars

2021 ◽  
Vol 17 (4) ◽  
pp. 74-82
Author(s):  
Paula Beatriz Ramos Guimarães ◽  
Mayara de Oliveira Vidotto Figueiredo ◽  
Tiago Benedito dos Santos ◽  
Alessandra Ferreira Ribas
Keyword(s):  
Salt Stress ◽  
Molecular Mechanisms ◽  
Culture Media ◽  
Proline Content ◽  
Quantitative Expression ◽  
Mechanisms Of Adaptation ◽  
The Media ◽  
Urochloa Brizantha ◽  
Biomass Reduction

Urochloa brizantha is the main forage grass to raise cattle in Brazil, but salt stress can reduce yield. Physiological and molecular mechanisms of adaptation to salt stress remain poorly understood in this species. The objective of this study was to evaluate the responses of three cultivars of U. brizantha to in vitro salt stress. Seeds of three cultivars (Piatã, Marandu, and Xaraés) germinated in filter paper and then transferred to growth on culture media in vitro containing 0, 50, 100, and 200 mg L-1 of sodium chloride (NaCl). Biometric parameters and proline content were determined after 28 days. The data were subjected to analysis of variance and the separation of means was performed by the LSD test (p<0.05). Semi-quantitative expression of the Δ1-pyrroline-5-carboxylate synthase (P5CS1) gene was performed. In all cultivars, increase of NaCl concentration in the media affected roots and shoots growth. Xaraes cultivar presented the greater biomass reduction while Marandu cultivar was the least affected. Salt stress increased by approximated 0.6 folds transcription of the P5CS1 gene in all cultivars. However, Marandu cultivar presented a higher proline content and least biomass reduction suggesting a better response to in vitro to salt stress.

scholarly journals In Vitro Effect of Crude Extract from Traganum Nudatum on Glucose-Uptake in Liver Slices Isolated from Westar Rats

2020 ◽  
Vol 4 (2) ◽  
pp. 550-551
Author(s):  
Faiza Mouderas
Keyword(s):  
Glucose Uptake ◽  
Crude Extract ◽  
Wistar Rats ◽  
Culture Media ◽  
Chronic Hyperglycemia ◽  
Liver Slices ◽  
Insulin Sensitizers ◽  
The Media ◽  
Vitro Effect

Background: Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia resulting from defects in insulin secretion, insulin action, or both. There are many classes of drugs used for treatment, and these include insulin sensitizers, insulin secretagogues, and agents that delay the absorption of carbohydrates from the bowel. This study intends to investigate the effect of crude extract from a plant from South Algeria Traganum nudatum (Chenopodiaceae) on glucose uptake in liver slices isolated from Wistar rats. Methods: The liver slices were incubated for 90 min at 37° in normoglycaemic (1g/l of glucose) and hyperglycaemic (3g/l of glucose) KRBA Krebs Ringer Bicarbonate Albumin 4% media using 24 well-polyethylene plates. In each, well different concentrations of insulin (10, 50 and 100µU/ml) and hydromethanolic crude extract (100, 200 and 500µg/ml) were added. After every 30 minutes, aliquots of the culture media were assayed for the determination of glucose left. Results: Tests showed that the glucose left after 90 minutes in the media which contained insulin at 100µg/ml was the lowest (0.44 and 1.41 )g/l in the normo and hyperglycaemic media respectively, which reflect that insulin at this concentration was the most effective on the stimulation of glucose uptake. The extract had the highest effect at 500µg/ml, the concentrations of glucose left after 90 minutes of incubation were found to be (0.38 and 1.31)g/l in the normoglycaemic and hyperglycaemic media respectively. Conclusion: From the obtained results, it can be concluded that our extract seems to have an insulin-like effect on glucose uptake in liver slices isolated from Wistar rats.

scholarly journals In Vitro Multiplication of Potato (Solanum tuberosum L.) cv. Cristal under Different MS Salt and Sucrose Concentrations

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 471E-471
Author(s):  
Gerson R. de L. Fortes ◽  
Luciana B. Andrade ◽  
Marisa de F. Oliveira ◽  
Nilvane T.G. Müller ◽  
Janine T. C. Faria
Keyword(s):  
Culture Media ◽  
Solanum Tuberosum L ◽  
Sucrose Concentration ◽  
Sugar Content ◽  
Potato Cultivar ◽  
In Vitro Multiplication ◽  
Full Strength ◽  
High Dry Matter ◽  
The Media

The potato cultivar Cristal has recently been released by the CPACT/EMBRAPA Breeding Program. Such cultivar was selected for having high dry matter and low sugar content, which makes it desirable for the chip industry. However, this is a recalcitrant cultivar as far as in vitro multiplication is concerned. The aim of this work was to improve the rate of multiplication for this cultivar when it was submitted to different MS salt and sucrose concentrations in the culture media. Two-bud microcuttings were inoculated in test tubes (20 × 150) mm with 10 ml MS media at 3/4-, 1/2-, and full-strength and MS vitamins added to: myo-inositol (100 mg·L–1), agar (7.0 g·L–1) and sucrose as follows: 10, 20 and 30 g·L-1. Each treatment was repeated eight times and each replicate had eight explants. After inoculation the whole material was kept in a growth room at 25 ± 2°C, 16-hr photoperiod and 2000 lux. The evaluation was done 35 days later. It was found and increase in the number of buds as the sucrose concentration in the media decreased. As far as MS salts are concerned no difference in bud number was observed. The rate of multiplication was slightly higher for MS media at full strength and sucrose at low concentration (10 g·L–1). This treatment could be recommended for this cultivar.

scholarly journals Efisiensi Mikropropagasi Pisang Kepok Amorang melalui Modifikasi Formula Media dan Temperatur

2016 ◽  
Vol 6 (2) ◽  
pp. 91
Author(s):  
Yati Supriati
Keyword(s):  
In Vitro Culture ◽  
Incubation Temperature ◽  
Culture Media ◽  
Shoot Multiplication ◽  
Root Induction ◽  
Incubation Condition ◽  
Growth Environment ◽  
Media Formulation ◽  
The Media

<p>Micropropagation Efficiency of Banana cv Kepok<br />Amorang through Modifications of Culture Media and<br />Incubation Temperature. Yati Supriati. The budless<br />banana cv Kepok Amorang is potentially commercialized<br />due to its sweet taste and does not have flower bud, hence<br />reduced the potential of being infected by the blood disease<br />pathogen. Enhancement of banana industry needs continuous<br />supplies of large number banana seedlings. In vitro<br />culture enable the production of seedlings in a large scale,<br />uniform, quick. The research aims: (1) to formulate an<br />efficient medium for in vitro multiplication of cv Kepok<br />Amorang shoot, (2) to identify efficient growth environment<br />for in vitro culture of cv Kepok Amorang, and (3) to formulate<br />an efficient culture medium for roots inductions of cv<br />Kepok Amorang. The plant material used was in vitro culture<br />of Kepok cv Amorang, 2 cm in height without leaf and root.<br />The media formulation for shoot multiplication were full<br />strength, half strength, one fourth strength MS media,<br />supplemented with either 1, 3, or 5 ppm IBA. On optimization<br />step, the media tested were MS, Knop, Knop and<br />Heller, Hyponex N, Growmore N, and Rosasol N containing<br />of 1 ppm BA. The explants were incubated in culture room<br />with 8, 12, and 16 hours photoperiod with temperatures 30oC<br />(non air conditioned) and 25oC (air conditioned). The root<br />induction trial was done using MS, Knop, Knop and Heller,<br />Hyponex N, Growmore N, and Rosasol N media containing<br />of 1 ppm and 3 ppm IBA. The results showed that the best<br />medium formula for shoot multiplication was ¼ MS + 1 ppm<br />IBA. The best incubation condition was 16 hours photoperiods<br />at 30oC. The best media for root induction was<br />Hyponex 2 g/l + 1 ppm IBA. This culture method reduced<br />cost by Rp 261.7 per plantlet through efficiency of media<br />formulation and electricity use.</p>

403. Differential effect of hexoses on sperm metabolism and function in culture

2008 ◽  
Vol 20 (9) ◽  
pp. 83
Author(s):  
H. W. Bakos ◽  
M. Lane
Keyword(s):  
Lipid Peroxidation ◽  
Dna Damage ◽  
Carbohydrate Metabolism ◽  
Culture Media ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Sperm Function ◽  
Ros Production ◽  
Sperm Dna Damage ◽  
The Media

Currently there is lack information regarding how human spermatozoa regulate their energy metabolism. This is surprising considering that carbohydrate metabolism is a vital point for the understanding of sperm function. This coupled with the increased use of assisted reproductive technology and the importance of a well balanced culture media has led us to hypothesise that an imbalance of carbohydrate presence in the media may alter sperm function, particularly in relation to oxidative stress, DNA damage and lipid peroxidation. Sperm samples were obtained from three healthy normospermic donors for this study. Motile sperm were separated from semen samples using density gradient separation. Samples were incubated at different media conditions with varying glucose or fructose concentrations (0, 2.5, 25 mM) for 6–24hrs. Reactive oxygen species (ROS) were measured using 5-(and-6)-carboxy-2', 7'-dichlorofluorescein diacetate (DCFDA). Sperm DNA damage was determined using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling (TUNEL). Lipid peroxidation was assessed using the probe BODIPY (581/591) C11. Carbohydrate uptake from the media was measured using a fluorometric procedure. Statistical differences between treatments were assessed by ANOVA and Bonferroni post-hoc test. No significant motility differences were found following treatments. Results showed an increased level of ROS production as glucose concentration increased (P < 0.05). This was accompanied by an increased number of TUNEL positive cells (P < 0.05). Furthermore, lipid peroxidation of spermatozoa was significantly increased when incubated under high glucose concentrations (P < 0.01). In contrast, increases in fructose concentrations did not alter ROS levels or the number of TUNEL positive cells. Sperm metabolised both glucose and fructose in vitro and the removal of one carbohydrate resulted in a compensatory increase in the metabolism of the other. To our knowledge, this is the first report providing evidence that altered carbohydrate metabolism may induce ROS production, lipid peroxidation and increase the number of sperm exhibiting DNA damage.

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