512. COMMERCIAL EQUINE CHORIONIC GONADOTROPHIN ISOFORM COMPOSITION, IMMUNOACTIVITY, AND BIOACTIVITY IN A MURINE MODEL USING OVARIAN AUGMENTATION AND TESTICULAR INTERSTITIAL CELL BIOASSAYS

2009 ◽  
Vol 21 (9) ◽  
pp. 111 ◽  
Author(s):  
U. A. Ciller ◽  
J. D. McFarlane ◽  
J. R. McFarlane
Keyword(s):  
Interstitial Cell ◽  
Chorionic Gonadotrophin ◽  
In Vitro Bioassay ◽  
Glycoprotein Hormone ◽  
Immature Female ◽  
Future Studies ◽  
In Vivo Bioassay

Equine chorionic gonadotrophin (eCG) is a placental glycoprotein hormone that is harvested from the plasma of pregnant mares for the formulation of commercial products which are used in a variety of assisted reproductive procedures in livestock. It is well documented that the bioactivity of gonadotrophin products is highly variable. The aim of this study was therefore to determine how different eCG products affected target tissues and cells. Isoforms of eCG were separated with iso-electric focusing and immunoactivity measured with RIA. For in vivo bioassay, dose-response eCG treatments were administered subcutaneously to immature female mice with follow-up treatment on day 2. On day 3 the mice were asphyxiated and ovaries dissected and weighed relative to standard. For in vitro bioassay, adult male mice were asphyxiated, testes removed, decapsulated, dispersed, strained and washed with DMEM:F12 0.1% BSA. The cell stock was counted and diluted for culture at 20,000 cells/well. Cell viability was determined using trypan blue. Five doses were tested for each eCG product. The cells were incubated at 32ºC for 3 hours in 5.0% CO2 in humidified conditions. Media was collected after 3 hours and immediately assayed for testosterone with RIA. Product eCG with ~90% isoforms with pH 3.0–3.7 and ~10% with pH 3.8–4.4, showed 29% greater biologically activity in the ovarian augmentation assay and 44.8% more immunoactivity then stated product bioactivity. Product eCG with ~70% isoforms with pH 3.0–3.7 and ~30% with pH 3.8–7.4 was 3.0% less effective in vivo and had 7.9% less immunoactivity then stated bioactivity, however, in vitro testosterone production was more effectively stimulated then with the previous more acidic eCG product. Our study shows a selective difference in commercial eCG biological activity that appears to depend on subtle isoform heterogeneity. Future studies aim to determine the specific actions of eCG isoforms in target tissues.

The International Standard for Pituitary FSH: Collaborative study of the Standard and of four other purified human FSH preparations of differing molecular composition by bioassays, receptor assays and different immunoassay systems

1989 ◽  
Vol 123 (2) ◽  
pp. 275-293 ◽  
Author(s):  
P. L. Storring ◽  
Gaines Das R. E.
Keyword(s):  
Geometric Mean ◽  
In Vitro Bioassay ◽  
International Standard ◽  
Receptor Assay ◽  
Specific Activities ◽  
In Vivo Bioassay ◽  
Ranking Order ◽  
Receptor Assays

ABSTRACT The International Standard for Pituitary FSH (IS; in ampoules coded 83/575) was assayed in terms of the Second International Reference Preparation of Human Pituitary FSH and LH for Bioassay (IRP 78/549) by 27 laboratories in 13 countries using bioassays, receptor assays and immunoassays. Estimates of the FSH content of the IS by in-vivo bioassay were homogeneous both within and between laboratories and gave a combined geometric mean (with 95% fiducial limits) of 79·9 (74·6–85·4) i.u./ampoule. Estimates by different in-vitro bioassays and receptor assays were also homogeneous between assays and laboratories, and gave a combined geometric mean (with 95% fiducial limits) of 31·2 (28·8–33·9) i.u./ampoule. However, estimates by the 19 different immunoassay systems were heterogeneous and varied between 5 and 31 i.u./ampoule. The material in ampoules coded 83/575 was established by the World Health Organization as the International Standard for Pituitary FSH. It was assigned a unitage of 80 i.u./ampoule on the basis of its calibration by in-vivo bioassay, because this assay best identifies and defines the hormone. However, the introduction of the new IS will necessitate the recalibration of immunoassay kits. FSH 84/530, prepared in the same way as the IS from the same FSH preparation, did not differ significantly from the IS in any of the assay systems studied and appeared to be equally suitable as a standard. Four highly purified preparations of human FSH (FSH A–D), differing in their isoform compositions and in their in-vivo: in-vitro bioactivity ratios, were also studied. The ranking order of the specific activities of FSH A–D by in-vitro bioassays paralleled their order by receptor assays and the order of their content of FSH isoforms with isoelectric points > 4·5. (Potency estimates of FSH B and C in terms of the IS were greater by receptor assay than by in-vitro bioassay.) The overall ranking order of the specific activities of FSH A–D by immunoassays was different. Contrary to expectation, estimates in terms of the IS of specific activities by immunoassay differed more between preparations than those by in-vitro bioassay or receptor assay. Differences in specificity between immunoassay systems were demonstrated not only in the calibration of the IS in terms of the crude FSH of IRP 78/549 but also in the comparisons of the highly purified FSH in the IS and FSH A–D. The differences in the immunoreactivities and bioactivities of FSH preparations differing in their isoform compositions greatly complicate the standardization of assays for FSH. Journal of Endocrinology (1989) 123, 275–293

A COMPARISON OF PREPARATIONS OF HIGHLY PURIFIED HUMAN PITUITARY FOLLICLE-STIMULATING HORMONE: DIFFERENCES IN THE FOLLICLE-STIMULATING HORMONE POTENCIES AS DETERMINED BY IN-VIVO BIOASSAY, IN-VITRO BIOASSAY AND IMMUNOASSAY

1981 ◽  
Vol 91 (2) ◽  
pp. 353-362 ◽  
Author(s):  
P. L. STORRING ◽  
A. A. ZAIDI ◽  
Y. G. MISTRY ◽  
BERIT FRÖYSA ◽  
BRIDGET E. STENNING ◽  
...  
Keyword(s):  
Biological Activity ◽  
Follicle Stimulating Hormone ◽  
In Vitro Bioassay ◽  
Human Pituitary ◽  
International Reference ◽  
Biological Potency ◽  
Reference Preparation ◽  
In Vivo Bioassay

The FSH potencies of 12 preparations of highly purified human pituitary FSH, originating from six different laboratories, were determined by in-vivo and in-vitro bioassays and by immunoassay in terms of the First International Reference Preparation of Human Pituitary Gonadotrophins (FSH and LH) for Bioassay (IRP; coded 69/104). The contamination of these FSH preparations with LH was also determined. Estimates of protein content were based on the absorbance at 280 nm of solutions of the preparations, assuming that A1%1 cm 280 = 10. The FSH potencies varied between different preparations from 827 i.u./mg to 13 100 i.u./mg by in-vivo bioassay; from 2930 to 14 600 i.u./mg by in-vitro bioassay and from 1680 to 5690 i.u./mg by immunoassay. The ratios of in-vivo biological activity relative to in-vitro biological activity and to immunoreactivity respectively varied between preparations from 0·06 to 2·3 and from 0·15 to 4·1, and there was a significant positive correlation between each of these ratios and the in-vivo biological potency of the preparations; such differences could be due to varying degrees of sialylation between preparations. On the other hand the ratios of in-vitro biological activity to immunoreactivity between preparations were fairly constant (approx. 2). The excess biological activity relative to immunoreactivity observed, in terms of the IRP, in all these materials is consistent with recent findings of some immunoreactive FSH in the IRP unassociated with biological activity. These data did not demonstrate any significant advantage, in terms of FSH in-vivo biological potency, from the use of fresh-frozen rather than acetone-dried pituitary glands for the isolation of FSH. Contamination of all these preparations with LH appeared to be less than 3% (w/w), as determined by in-vitro bioassay and by immunoassay. The results of this study are discussed in relation to the selection of material for an international reference preparation for immunoassay and attention is drawn to the value of high in-vivo biological FSH potency as a criterion of the identity of a preparation as well as of its freedom from contaminants without FSH biological activity.

A comparison of preparations of highly purified human pituitary luteinizing hormone: differences in the luteinizing hormone potencies as determined by in vivo bioassays, in vitro bioassay and immunoassay

1982 ◽  
Vol 101 (3) ◽  
pp. 339-347 ◽  
Author(s):  
P. L. Storring ◽  
A. A. Zaidi ◽  
Y. G. Mistry ◽  
Monica Lindberg ◽  
Bridget E. Stenning ◽  
...  
Keyword(s):  
Luteinizing Hormone ◽  
In Vitro Bioactivity ◽  
In Vitro Bioassay ◽  
Response Curves ◽  
Human Pituitary ◽  
International Reference ◽  
Reference Preparation ◽  
In Vivo Bioassay

Abstract. The LH potencies of 12 preparations of highly purified human pituitary LH, from 6 laboratories, were estimated by 2 in vivo bioassays and an in vitro bioassay in terms of the International Reference Preparation of Human Pituitary Gonadotrophins (FSH and LH) for Bioassay (coded 69/104); and by immunoassay in terms of the International Reference Preparation of Human Pituitary Luteinizing Hormone for Immunoassay (IRP; coded 68/40). The LH potencies varied between preparations, including the IRP (68/40), from 864 to 5740 IU/mg by seminal vesicle weight gain (SVW) assay; from 1510 to 11500 IU/mg by ovarian ascorbate depletion (OAAD) assay; from 4490 to 14500 IU/mg by in vitro (testicular interstitial-cell testosterone production) bioassay; and from 2030 to 9180 IU/mg by immunoassay. Estimates of protein content were based on the assumption that the absorbance of LH at 280 nm (A 1% 1 cm) was 6.0. The LH potency of most preparations was highest by in vitro bioassay and lowest by SVW assay. The correlation between activities determined by SVW and OAAD assays was more marked than that between estimates by OAAD assay and in vitro bioassay; there was no correlation between estimates by SVW assay and in vitro bioassay. The slopes of the log dose-response curves of preparations in the OAAD assay were positively correlated with their potencies by OAAD assay and negatively correlated with the slopes of their log dose-response curves in the SVW assay. The qualitative differences between preparations are considered to be a reflection of the heterogeneity of LH and of its modification by different purification procedures. The present data, together with the different patterns of heterogeneity found in some of these preparations by isoelectric focusing in a separate study, suggest that the more basic molecular forms of LH, which are preferentially purified during the isolation of LH free from FSH and TSH, have shorter plasma survival times than the more acidic forms. The LH immunoreactivities of all preparations were significantly correlated with their potencies estimated by each of the in vivo bioassays but not with those estimated by in vitro bioassay. The ratios of in vitro bioactivity (in terms of IRP (68/40)): immunoreactivity varied between preparations from 0.53–1.5. The FSH content of each preparation was less than 2% (w/w) by bioassay and immunoassay. Most preparations were more potent by in vitro bioassay than by in vivo bioassay, which contrasted with, and complemented, findings for purified FSH preparations. This indicated that, as in the case of LH, the more basic molecular species of FSH are associated with lower ratios of in vivo: in vitro bioactivity than are the more acidic species. This study provides the most comprehensive comparison available of the activities of purified preparations of LH isolated from frozen and acetone-dried human pituitary glands in different experienced laboratories. These data are needed for selecting material for an international reference preparation of LH for immunoassay on the basis of high LH potency by in vivo bioassay, recommended by the WHO as a criterion for the identity of the hormone and for its freedom from contaminants. The consequences of the heterogeneity of LH are considered for the purification of the reference material and for the suitability of the latter for the various types of specimens which require LH assays.

TESTICULAR ANDROGEN BIOSYNTHESIS FOLLOWING CORTICOTROPHIN AND HUMAN CHORIONIC GONADOTROPHIN ADMINISTRATION

1965 ◽  
Vol 50 (3) ◽  
pp. 365-378 ◽  
Author(s):  
Edgar J. Schoen ◽  
Leo T. Samuels
Keyword(s):  
Interstitial Cell ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Steroid Biosynthesis ◽  
Clear Cut ◽  
Adrenocortical Hyperplasia ◽  
Testicular Interstitial Cells ◽  
Stimulation Of

ABSTRACT The effects of in vivo administration of corticotrophin (ACTH) and human chorionic gonadotrophin (HCG) on rat testes were studied. HCG, administered for 14 days to young rats, led to testicular interstitial cell hypertrophy and hyperplasia, accompanied by stimulation of in vitro testicular androgen production, as manifested by increased 17α-hydroxylase and 17β-desmolase enzyme activities, with no clear-cut effect on 20α-hydroxysteroid dehydrogenase activity. The findings in ACTH-treated animals were similar to those in the saline-treated controls. The lack of influence of ACTH on the testicular enzymes involved in steroid biosynthesis is consonant with the concept that ACTH-dependent bilateral testicular tumours occurring in boys with congenital adrenocortical hyperplasia arise from »adrenocortical rest cells« rather than from testicular interstitial cells.

Synthesis and Fungicidal Activity of (E)-5-[1-(4-Phenyl-2-oxo-1-oxaspiro[4.5]dec-3-en-3-yl)ethylidene]-2-aminoimidazolin-4-one Derivatives

Synthesis ◽  
2017 ◽  
Vol 49 (20) ◽  
pp. 4663-4669
Author(s):  
Zhao Yu ◽  
Tang Bo ◽  
Guan Aiying ◽  
Wang Weiwei ◽  
Zhang Zhenhua ◽  
...  
Keyword(s):  
1H Nmr ◽  
Fungicidal Activity ◽  
In Vitro Bioassay ◽  
In Vivo Bioassay

(E)-5-[1-(4-Phenyl-2-oxo-1-oxaspiro[4.5]dec-3-en-3-yl)eth­yl­idene]-2-aminoimidazolin-4-one derivatives, as novel fungicidal agents, are designed and synthesized in moderate to excellent yields in four steps from (1-hydroxycyclohexyl)(phenyl)methanone and diketene as the starting materials. The products are characterized by 1H NMR and HRMS (ESI) analysis. An in vivo bioassay shows that some of the products exhibit good to excellent inhibition against P. cubensis and C. lagenarium, whilst up to 94.7% inhibition against P. capsici and up to 78.1% inhibition against B. cinerea is demonstrated in the in vitro bioassay. EC50 values of 3.40 and 5.86 μg/mL are demonstrated against P. capsici and B. cinerea.

scholarly journals TRIM25 regulates oxaliplatin resistance in colorectal cancer by promoting EZH2 stability

2021 ◽  
Vol 12 (5) ◽  
Author(s):  
Sha Zhou ◽  
Jianhong Peng ◽  
Liuniu Xiao ◽  
Caixia Zhou ◽  
Yujing Fang ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Disease Free Survival ◽  
Free Survival ◽  
Epigenetic Regulator ◽  
Crc Management ◽  
Disease Free ◽  
Resistance To Chemotherapy

AbstractResistance to chemotherapy remains the major cause of treatment failure in patients with colorectal cancer (CRC). Here, we identified TRIM25 as an epigenetic regulator of oxaliplatin (OXA) resistance in CRC. The level of TRIM25 in OXA-resistant patients who experienced recurrence during the follow-up period was significantly higher than in those who had no recurrence. Patients with high expression of TRIM25 had a significantly higher recurrence rate and worse disease-free survival than those with low TRIM25 expression. Downregulation of TRIM25 dramatically inhibited, while overexpression of TRIM25 increased, CRC cell survival after OXA treatment. In addition, TRIM25 promoted the stem cell properties of CRC cells both in vitro and in vivo. Importantly, we demonstrated that TRIM25 inhibited the binding of E3 ubiquitin ligase TRAF6 to EZH2, thus stabilizing and upregulating EZH2, and promoting OXA resistance. Our study contributes to a better understanding of OXA resistance and indicates that inhibitors against TRIM25 might be an excellent strategy for CRC management in clinical practice.

scholarly journals Differential Normalization of Mucosal Interleukin-8 and Interleukin-6 Activity after Helicobacter pyloriEradication

1998 ◽  
Vol 66 (10) ◽  
pp. 4742-4747 ◽  
Author(s):  
Takafumi Ando ◽  
Kazuo Kusugami ◽  
Masahiro Ohsuga ◽  
Kenji Ina ◽  
Masataka Shinoda ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Eradication Therapy ◽  
Interleukin 8 ◽  
Biopsy Specimens ◽  
Mucosal Tissues ◽  
H Pylori ◽  
Triple Treatment

ABSTRACT There is differential resolution of mucosal infiltration with neutrophils and mononuclear cells following successfulHelicobacter pylori eradication. We investigated the effects of H. pylori eradication on mucosal interleukin-8 (IL-8) and IL-6 activity in relation to the resolution of H. pylori-associated gastritis. Eighty-one duodenal ulcer patients with H. pyloriinfection received dual- or triple-treatment eradication therapy, and mucosal biopsy specimens obtained at the initial and follow-up endoscopic examinations were cultured in vitro for 24 h. The levels of IL-8 and IL-6 were measured by enzyme-linked immunosorbent assays. In the 42 patients in whomH. pylori eradication failed, there was little change in the numbers of neutrophils and mononuclear cells infiltrating the mucosa and in IL-8 and IL-6 activity. In the 39 patients in whom H. pylori was eradicated, there was normalization both in the numbers of infiltrating neutrophils and in mucosal IL-8 activity, which was evident within 1 month following therapy. In contrast, there was a gradual resolution of mononuclear cell infiltration over a 6-month period, accompanied by a gradual normalization in IL-6 levels. Addition of H. pylori to cultures of mucosal tissues induced a significant increase in IL-8 activity in both uninfected control subjects and patients from whom H. pylori was eradicated. However, this introduction yielded a significant increase in IL-6 activity only in the latter group. This study indicates a dichotomy in the changes of mucosal IL-8 and IL-6 activity afterH. pylori eradication. The rapid normalization of IL-8 after H. pylori eradication and the ability of H. pylori cells to stimulate IL-8 in control tissues indicate that IL-8 induction is a part of the innate (nonimmune) responses to this organism. In contrast, the results of experiments analyzing IL-6 activity in cultured mucosal tissues suggest that the gradual resolution of mucosal IL-6 activity and mononuclear infiltration after successful eradication observed in vivo may reflect gradually diminishing residual immune responses against H. pylori.

Antimalaria Activity of Cassia spectabilis DC Leaf and Its Inhibition Effect in Heme Detoxification

2020 ◽  
Author(s):  
Wiwied - Ekasari ◽  
Dewi Resty Basuki ◽  
Heny - Arwati ◽  
Tutik Sri Wahy
Keyword(s):  
Antimalarial Activity ◽  
Ethanol Extract ◽  
Ic50 Value ◽  
Cassia Spectabilis ◽  
In Vivo Testing ◽  
Chloroquine Diphosphate ◽  
Better Than

Abstract Background In previous studies, Cassia spectabilis DC leaf has shown a good antimalarial activity. Therefore, this study is a follow-up study of leaf activity and mechanism of C. spectabilis DC as an antimalarial. Methods In vitro antimalarial activity testing using P. falciparum which was done with bioassay guide isolation in order to obtain the active compound. In vivo testing towards infected P. berghei mice was conducted to determine the effects of antimalarial prophylaxis and antimalarial activity in combination with artesunate. Whereas, heme detoxification inhibition testing as one of the antimalarial mechanisms was carried out using the Basilico method. Results The results showed that active antimalarial isolate obtained from C. spectabilis DC leaf had a structural pattern that was identical to (-)-7-hydroxyspectaline. Prophylactic test on infected P. berghei mice obtained the highest dose of inhibition percentage of 90% ethanol extract of C. spectabilis DC leaf was 68.61% while positive (doxycycline) control at 100 mg kg-1 was 73.54%. In antimalarial testing in combination with artesunate, it was found that administering 150 mg kg-1 (three times a day) of C. spectabilis DC (D0 − D2) + artesunate (D2) was better than the standard combination of amodiaquine + artesunate with 99.18% and 92.88% inhibition percentage. For the inhibitory activity of heme detoxification from ethanol extract 90%, C. spectabilis DC leaf had IC50 value of 0.375 mg mL-1 which was better than chloroquine diphosphate. Conclusion These results showed that C. spectabilis DC leaves possesses potent antimalarial activity and may offer a potential agent for effective and affordable antimalarial phytomedicine.

scholarly journals Model-Based Roentgen Stereophotogrammetric Analysis to Monitor the Head–Taper Junction in Total Hip Arthroplasty in Vivo—And They Do Move

Materials ◽  
2020 ◽  
Vol 13 (7) ◽  
pp. 1543 ◽  
Author(s):  
Jing Xu ◽  
Robert Sonntag ◽  
J. Philippe Kretzer ◽  
Dominic Taylor ◽  
Raimund Forst ◽  
...  
Keyword(s):  
Femoral Stem ◽  
Geometrical Shape ◽  
Roentgen Stereophotogrammetric Analysis ◽  
Model Based ◽  
Similar Range ◽  
Standard Marker ◽  
Relative Migration

Model-based Roentgen stereophotogrammetric analysis (RSA) using elementary geometrical shape (EGS) models allows migration measurement of implants without the necessity of additional attached implant markers. The aims of this study were: (i) to assess the possibility of measuring potential head–taper movement in THA in vivo using model-based RSA and (ii) to prove the validity of measured head–taper migration data in vitro and in vivo. From a previous RSA study with a 10 years follow-up, retrospectively for n = 45 patients head–taper migration was calculated as the relative migration between femoral ball head and taper of the femoral stem using model-based RSA. A head–taper migration of 0.026 mm/year can be detected with available RSA technology. In vitro validation showed a total migration of 268 ± 11 µm along the taper axis in a similar range to what has been reported using the RSA method. In vivo, a proof for interchangeable applicability of model-based RSA (EGS) and standard marker-based RSA methods was indicated by a significant deviation within the migration result after 12-month follow-up for all translation measurements, which was significantly correlated to the measured head–taper migration (r from 0.40 to 0.67; p < 0.05). The results identified that model-based RSA (EGS) could be used to detect head–taper migration in vivo and the measured movement could be validated in vitro and in vivo as well. Those findings supported the possibility of applying RSA for helping evaluate the head–taper corrosion related failure (trunnionosis).

Immunochemical analysis of the human chorionic gonadotrophin-like material secreted by 'normal' and neoplastic urothelial cells

1989 ◽  
Vol 2 (2) ◽  
pp. 107-112 ◽  
Author(s):  
R. K. Iles ◽  
T. Chard
Keyword(s):  
Bladder Tumour ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Sds Page ◽  
Β Hcg ◽  
Bladder Carcinomas ◽  
Immunochemical Characteristics ◽  
Normal Urothelium

ABSTRACT Material with the immunochemical characteristics of human chorionic gonadotrophin (hCG) is produced by bladder tumour cells in vitro and in vivo. In order to characterize this material further, media were collected from 17 cell cultures (three choriocarcinomas, seven bladder carcinomas and seven 'normal' urothelium). The hCG-like material was compared with pregnancy hCG and purified α- and β-subunits by specific radioimmunoassays. Media were also submitted to affinity chromatography and the fractions further analysed by SDS-PAGE and Western blotting. It was shown that both the neoplastic and normal urothelium produced only free β-subunit-like material. This urothelial 'β-hCG' has the same molecular weight and electrophoretic mobility as that present in the intact hCG of pregnancy.

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