scholarly journals Involvement of sperm sulphatases in early sperm-zona interactions in the hamster

1985 ◽  
Vol 78 (1) ◽  
pp. 247-261
Author(s):  
K.K. Ahuja ◽  
D.J. Gilburt
Keyword(s):  
Zona Pellucida ◽  
Artificial Substrates ◽  
Dependent Manner ◽  
Vitro Fertilization ◽  
Pre Treatment ◽  
High Concentrations ◽  
Dose Dependent ◽  
Fertilization System

Sulphatase preparations from Abalone entrails, the limpet Patella vulgata and ox liver, as well as artificial substrates for these enzymes, were used in the hamster in vitro fertilization system to study the possible roles of sperm sulphatases in sperm-zona pellucida interactions. p-nitrophenyl sulphate, p-nitrocatechol sulphate, ascorbate 2-sulphate, as well as D-glucopyranose and D-galactopyranose, both sulphated at the 3, 4 or 6 position but not the 2 position, inhibited fertilization in a dose-dependent manner. Sperm-egg fusion was not inhibited by the substrates used and eggs pre-treated with sulphates could readily be fertilized. Sperm motility and therefore viability was unaffected by inhibitory concentrations of substrates as determined by rhodamine 123 labelling of motile spermatozoa. Acrosomal integrity of rhodamine-labelled spermatozoa was assessed and found to be unaffected by inhibitory levels of substrates. Fertilization was inhibited by high concentrations of the two molluscan sulphatases but not by purified ox liver sulphatase. Pre-treatment of eggs with these enzymes did not prevent fertilization. Long-term exposure of hamster oocytes to N-acetylhexosaminidase or limpet sulphatase caused thinning and distension of the zona pellucida but these changes were not observed with the ox liver sulphatase. The results suggest that a glycosulphatase is probably released from hamster spermatozoa during sperm-egg adhesion and, or, penetration. If sperm-egg adhesion molecules are sulphated, the commercially available sulphatases would be unsuitable for their characterization.

Differences between antigenic determinants of pig and cat zona pellucida proteins

Reproduction ◽  
2000 ◽  
pp. 15-23 ◽  
Author(s):  
K Jewgenow ◽  
M Rohleder ◽  
I Wegner
Keyword(s):  
In Vitro Fertilization ◽  
Zona Pellucida ◽  
Antigenic Determinants ◽  
Vitro Fertilization ◽  
Contraceptive Vaccine ◽  
Sperm Binding ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Dose Dependent

Despite many efforts, the control of reproduction in feral cat populations is still a problem in urban regions around the world. Immunocontraception is a promising approach; thus the present study examined the suitability of the widely used pig zona pellucida proteins (pZP) for contraception in feral domestic cats. Purified zona pellucida proteins obtained from pig and cat ovaries were used to produce highly specific antisera in rabbits. Antibodies against pZP raised in rabbits or lions were not effective inhibitors of either in vitro sperm binding (cat spermatozoa to cat oocytes) or in vitro fertilization in cats, whereas antibodies against feline zona pellucida proteins (fZP) raised in rabbits showed a dose-dependent inhibition of in vitro fertilization. Immunoelectrophoresis, ELISA and immunohistology of ovaries confirmed these results, showing crossreactivity of anti-fZP sera to fZP and to a lesser extent to pZP, but no interaction of anti-pZP sera with fZP. It is concluded that cat and pig zonae pellucidae express a very small number of shared antigenic determinants, making the use of pZP vaccine in cats questionable. A contraceptive vaccine based on feline zona pellucida determinants will be a better choice for the control of reproduction in feral cats if immunogenity can be achieved.

scholarly journals Ethnopharmacological basis for antispasmodic, antidiarrheal and antiemetic activities of Ceratonia siliqua pods

2017 ◽  
Vol 12 (4) ◽  
pp. 384
Author(s):  
Irfan Hamid ◽  
Khalid Hussain Janbaz
Keyword(s):  
Crude Extract ◽  
Video Clip ◽  
Dependent Manner ◽  
Ceratonia Siliqua ◽  
Spasmolytic Activity ◽  
Rabbit Jejunum ◽  
Pre Treatment ◽  
Dose Dependent

<p class="Abstract">The study was conducted to provide the ethnopharmacological bases of the crude extract of seed pods of <em>Ceratonia siliqua</em> in the gastrointestinal spasm, diarrhea and emesis. In segregated rabbit jejunum, it showed dose-dependent (0.01-10 mg/mL) relaxation of spontaneous as well as carbachol (1 µM)-induced contraction. Pre-treatment of segregated rat ileum with <em>C. siliqua</em>, significantly (p&lt;0.0001) suppressed the carbachol (1 µM)-induced contraction similar to atropine (1 µM). These results indicated that <em>C. siliqua</em> possesses spasmolytic activity through possible blockage of muscarinic receptor in jejunum preparations. Furthermore, the crude extract inhibited the castor oil-induced diarrhea, charcoal meal propulsion in mice and copper sulfate-induced retches in chicks in a dose-dependent manner (100, 200, 300 mg/kg). These in vitro and in vivo results indicate that <em>C. siliqua</em> possesses the spasmolytic and antidiarrheal activities mediated possibly through blockage of muscarinic receptors. Thus, this study provides a rationale for its folkloric use.</p><p><strong>Video Clip of Methodology</strong>:</p><p>12 min 42 sec   <a href="https://www.youtube.com/v/BQGWdIZqpsY">Full Screen</a>   <a href="https://www.youtube.com/watch?v=BQGWdIZqpsY">Alternate</a></p>

Sodium Butyrate Pre-Treatment Enhance Differentiation of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) into Hepatocytes and Improve Liver Injury

2021 ◽  
Vol 21 ◽  
Author(s):  
Qiu-Yun Li ◽  
Juan Chen ◽  
Yong-Heng Luo ◽  
Wei Zhang ◽  
En-Hua Xiao
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Dependent Manner ◽  
Hepatic Differentiation ◽  
Marrow Mesenchymal Stem Cells ◽  
Specific Factors ◽  
Pre Treatment ◽  
Dose Dependent

Objective: The treatment of liver failure by stem cell transplantation has attracted growing interest. Herein, we aim to explore the role of sodium butyrate (NaB) in the hepatic differentiation of bone marrow mesenchymal stem cells (BM-MSCs) under liver-specific factors induction in vitro and vivo. Materials & Methods: We isolated BM-MSCs from the mononuclear cell fraction of rabbit bone marrow samples, and identified the cells by Immunophenotypic analysis. We investigated the effects of different concentrations and induction conditions. The histone deacetylase inhibitor NaB induced hepatic differentiation of BM-MSCs under liver-specific factors induction in vitro. Morphological features, liver-specific gene and protein expression, and functional analyses in vitro and vivo were performed to evaluate the hepatic differentiation of BM-MSCs. Results: Our results showed that pre-treated NaB inhibited the expression of liver-specific protein in a dose-dependent manner. The induction efficiency of NaB with 24h pre-treatment was higher than that of NaB continuous intervention. 0.5 mM 24h NaB pre-treated cells can improve liver tissue damage in vivo. And the liver ALB, AAT and the serum TP were significantly increased, while the serum ALT was significantly reduced. Conclusion: Continuous NaB treatment can inhibit BM-MSCs proliferation in a dose-dependent manner at a certain concentration range. 0.5 mM 24h pre-treatment of NaB enhanced differentiation of BM-MSCs into hepatocytes and improves liver injury in vitro and vivo.

Impact of gonadotropins on oocyte maturation, fertilisation and developmental competence in vitro

2014 ◽  
Vol 26 (5) ◽  
pp. 752 ◽  
Author(s):  
Xuemei Wang ◽  
Tony Tsai ◽  
Jie Qiao ◽  
Zhan Zhang ◽  
Huai L. Feng
Keyword(s):  
Oocyte Maturation ◽  
Early Embryo ◽  
Developmental Competence ◽  
Dependent Manner ◽  
Success Rates ◽  
Maturation In Vitro ◽  
High Concentrations ◽  
Development In Vitro ◽  
Dose Dependent

The aim of the present study was to evaluate the dose-dependent effects of gonadotropins, either singly (Bravelle (B), Luveris (L), Menupur (M), Repronex (R), Gonal-F (G), Follism (F) and Norvarel (N)) or in combination (Menupur + Bravelle; Repronext + Bravelle; and Bravelle + Norvarel), on rates of oocyte maturation, fertilisation and early embryo development in vitro in an animal model. Bovine cumulus–oocyte complexes (COCs) were purchased commercially and cultured in TCM-199 with 10% fetal bovine serum supplemented with varying concentrations of gonadotropin (0, 5, 10, 20, 40 IU or United States Pharmacopoeia (USP) mL–1) for 24 and 48 h according to current IVF clinical stimulation protocols. All gonadotropins enhanced oocyte maturation in vitro in a dose-dependent manner. Individually, Gonal-F (Merck KGaA, Darmstadt, Germany), Follism (Merck Co, Whitehouse Station, NJ, USA) and Repronext (Ferring, Parsippany, NJ, USA) promoted oocyte maturation; in combination, they effectively enhanced COC expansion and increased the maturation competence of MII oocytes. However, high concentrations of gonadotropins may result in maturation arrest. Specific combinations of gonadotropins may change the rate of early embryonic development (8–16-cells) and morula–blastocyst formation. These data provide support for the responsiveness of bovine oocytes to gonadotropins in vitro and the need to consider variations in the relative concentrations and ratio of combinations (FSH/LH or human chorionic gonadotropin) for optimisation of oocyte developmental competence. The results of the present study could be applied to therapeutic clinical stimulation protocols and help improve IVF success rates.

Vitamin D as a Regulator of Adipocyte Differentiation Effects in vivo and in vitro

2018 ◽  
Vol 69 (3) ◽  
pp. 731-734
Author(s):  
Alin Constantin Pinzariu ◽  
Teodor Oboroceanu ◽  
Florin Zugun Eloae ◽  
Ioana Hristov ◽  
Victor Vlad Costan ◽  
...  
Keyword(s):  
Vitamin D ◽  
Adipocyte Differentiation ◽  
Dependent Manner ◽  
Omental Adipose Tissue ◽  
In Vivo Experiments ◽  
Dose Dependent ◽  
Stromal Stem Cells

The age-associated adiposity and the effect of long-term vitamin D was studied in vitamin D deficient rats. In in vivo experiments, the influence of a 9 months of vitamin D treatment (weekly oral gavage with 0.125 mg vitamin D3 (5000 IU)/100g body weight) on the adipocyte precursors from the omental adipose tissue was examinated. In in vitro experiment, rat adipose-derived mesenchymal stromal/stem cells (ASCs) were induced to differentiate into adipocytes in the presence or absence of 25(OH)D3 (0.25, 25, and 2500 nmol/L). ASCs derived from vitamin D-treated animals showed an increase adipogenic potential as compared to vitamin D-deficient rats. The addition of 25(OH)D3 inhibits the adipocyte differentiation and lipid deposition in a dose dependent manner.

High-Dose Calcitriol Modulates the Expression and Activity of Tissue Factor and Thrombomodulin in Tumor Cells.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 921-921
Author(s):  
Enriqueta Coll-Sangrona ◽  
Ali Amirkhosravi ◽  
Alshad S. Lalani ◽  
Liza Robles ◽  
Hina Desai ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Endothelial Cells ◽  
Tumor Cells ◽  
High Dose ◽  
Dependent Manner ◽  
High Concentrations ◽  
Dose Dependent ◽  
A549 Lung

Abstract Calcitriol, the hormonally-active metabolite of Vitamin D3, plays critical roles in calcium homeostasis, cell growth and differentiation, and immunoregulation. The anti-tumor activities of high-dose calcitriol have been demonstrated in a variety of preclinical models of solid tumors, leukemias and lymphomas. Recently, a new dose-intense formulation of calcitriol, termed DN-101 (Asentar™), was developed specifically for cancer therapy which allows for supraphysiological concentrations of calcitriol to be safely delivered in vivo to patients with cancer. In a recent Phase 2 clinical trial, DN-101 significantly increased overall survival and also reduced the incidence of thromboembolic events in men with androgen-independent prostate cancer receiving docetaxel-based chemotherapy. Based on previous observations we hypothesized that calcitriol’s anti-thrombotic effects in vivo may be due to the downregulation of Tissue Factor (TF) antigen and activity and/or upregulation of Thrombomodulin (TM). To test this hypothesis, we incubated A549 lung carcinoma, A375-C15 metastatic melanoma, THP-1 monocytic leukemia, and Eahy926 endothelial cells with increasing concentrations of calcitriol for 24 hrs. For TF induction, tumor cells were stimulated with TNFα for 5 hrs and activity was measured by a clotting assay and a thrombin generation assay (TGA). TM activity was measured by a chromogenic assay. TF and TM surface antigen were assessed by flow cytometry. Calcitriol prevented the induction of TF in TNFα-stimulated THP-1 cells in a dose-dependent manner (from 33% at 1 nM to 94% at 100 nM) as evidenced by a prolongation of plasma clotting time, a decrease in endogenous thrombin potential (ETP), and a reduction of surface TF antigen. In addition, the activity and surface expression of TM on THP-1 cells was increased significantly (40% and 3-fold respectively, P < 0.01) following 100 nM calcitriol treatment. Similarly, in TNFα-stimulated melanoma cells, calcitriol prevented the induction of TF activity (from 26% at 1 nM to 60% at 1 μM) and expression in a dose-dependent manner. High-dose calcitriol treatment also increased melanoma cell TM activity between 8% and 62%. In contrast, constitutively expressed TF activity and antigen were less affected by calcitriol in A549 lung carcinoma cells (12 to 28% reduction at concentrations between 1–100 nM) whilst TM activity and antigen were unaffected. In comparison to the tumor cells, calcitriol had no significant effect on TM or TF activity or antigen in TNFα-stimulated EAhy926 endothelial cells. In conclusion, we have demonstrated that high concentrations of calcitriol inhibit the induction of surface TF expression and upregulates TM in multiple tumor cell lines in vitro. The degree of the inhibition is proportional to the extent of TF induction by TNF-α. These in vitro results provide further support for the anticoagulant properties associated with high concentrations of calcitriol and may provide a rationale for understanding the lower incidence of thromboembolic complications observed in patients with metastatic prostate cancer treated with DN-101.

Histidine-rich basic peptide: a cardioactive neuropeptide from Aplysia neurons R3-14

1987 ◽  
Vol 57 (4) ◽  
pp. 1201-1209 ◽  
Author(s):  
J. T. Campanelli ◽  
R. H. Scheller
Keyword(s):  
Signal Sequence ◽  
Beat Frequency ◽  
Gel Filtration ◽  
Dependent Manner ◽  
Gene Encoding ◽  
Basic Peptide ◽  
High Concentrations ◽  
Dense Core Granules ◽  
Dose Dependent

We have previously demonstrated that neurons R3-14 of the Aplysia abdominal ganglia specifically express a gene encoding a 108-amino acid neuropeptide precursor. This precursor is postranslationally processed by cleavage of a signal sequence and two internal dibasic residues resulting in three peptides. The peptide products are colocalized in dense core granules throughout the R3-14 processes that innervate the efferent vein of the gill and the auricle. Gel filtration and reverse phase high-pressure liquid chromatography (rpHPLC) were used to purify a 4.9-kDa peptide produced by the R3-14 neurons. We call this peptide the histidine-rich basic peptide (HRBP), which reflects its primary structure. In vitro tension measurements of cannulated Aplysia hearts revealed dose-dependent cardioexcitatory actions of HRBP. HRBP increased both beat frequency and amplitude with a threshold of 10(-7) M. HRBP increased the amplitude of ventricular contractions in a dose-dependent manner, whereas the frequency of contraction is unaffected. In contrast both the amplitude and frequency of auricular contractions were enhanced. High concentrations of HRBP also had a positive tonotropic effect on the auricle. HRBP was also demonstrated to have actions on tissue of the gut. Circular muscles of the crop adjacent to the anterior gizzard showed infrequent spontaneous contractions. Both HRBP and acetylcholine (ACh) induced repetitive contractions of this muscle. Circular muscles of the posterior gizzard had a high degree of spontaneous activity when continually perfused. Contraction amplitude and frequency was increased by HRBP and ACh, whereas contractility was inhibited by Phe-Met-Arg-Phe-amide (FMRFamide).

Dissimilar peptidase production by avirulent and virulent promastigotes of Leishmania braziliensis: inference on the parasite proliferation and interaction with macrophages

Parasitology ◽  
2009 ◽  
Vol 136 (10) ◽  
pp. 1179-1191 ◽  
Author(s):  
A. K. C. LIMA ◽  
C. G. R. ELIAS ◽  
J. E. O. SOUZA ◽  
A. L. S. SANTOS ◽  
P. M. L. DUTRA
Keyword(s):  
Virulent Strain ◽  
Cellular Interaction ◽  
Leishmania Braziliensis ◽  
Avirulent Strain ◽  
Dependent Manner ◽  
Association Index ◽  
Cysteine Peptidase ◽  
Pre Treatment ◽  
Dose Dependent

SUMMARYIn the present paper, we have analysed the cellular and extracellular proteolytic activity profiles in 2 distinct Leishmania braziliensis strains: a recently isolated (virulent) and a laboratory-adapted (avirulent) strain. Quantitative and qualitative differences on the peptidase expression were observed in both strains. For instance, low-molecular mass acidic cysteine peptidase activities were detected exclusively in the virulent strain. Similarly, metallopeptidase activities were mainly produced by L. braziliensis virulent promastigotes. Interestingly, metallo- and cysteine peptidase activities were drastically reduced after several in vitro passages of the virulent strain. Western blotting, flow cytometry and fluorescence microscopy analyses were performed to detect homologous of the major leishmania metallopeptidase (gp63) and cysteine peptidase (cpb) in virulent and avirulent strains of L. braziliensis. Our results revealed that the virulent strain produced higher amounts of gp63 and cpb molecules, detected both in the surface and cytoplasm regions, than the avirulent counterpart. Metallo- (1,10-phenanthroline and EGTA) and cysteine peptidase (E-64) inhibitors arrested the growth of L. braziliensis virulent strain in a dose-dependent manner, as well as the association index with peritoneal murine macrophages. Conversely, these peptidase inhibitors did not affect either the proliferation or the cellular interaction of the avirulent strain. Corroborating these findings, the pre-treatment of the virulent strain with both anti-peptidase antibodies promoted a prominent reduction in the interaction with macrophages, while the association index of the avirulent strain to macrophage was only slightly diminished. Moreover, the spent culture medium from virulent strain significantly enhanced the association index between avirulent strain and macrophages, and this effect was reversed by 1,10-phenanthroline. Collectively, the results presented herein suggest that peptidases participate in several crucial processes of L. braziliensis.

scholarly journals Effects of zinc and molybdenum on European Bluestar (Amsonia orientalis): An in vitro study

2020 ◽  
Vol 4 (1) ◽  
pp. 32-41 ◽  
Author(s):  
Arda Acemi ◽  
Yonca Avcı Duman ◽  
Yonca Yüzügüllü Karakuş ◽  
Fazıl Özen
Keyword(s):  
In Vitro Study ◽  
Dependent Manner ◽  
Sod Activity ◽  
Metal Concentrations ◽  
Drought And Salt Stress ◽  
Toxic Concentrations ◽  
Dose Dependent ◽  
Zn Stress

AbstractThis study aimed to investigate the effects of possible zinc (Zn) and molybdenum (Mo) contaminations on the critically endangered European Bluestar (Amsonia orientalis). The effects of Zn and Mo were tested in a dose-dependent manner on in vitro cultures. Zn at 0.1 mM in the medium inhibited root development whereas Mo showed the same effect only at ≥2.5 mM concentration. Gradual inhibition of shoot development was observed after treatment with both metals. Protein contents were also negatively affected by increasing metal concentrations, while proline levels increased gradually. Successive increases in metal concentrations resulted in higher hydrogen peroxide (H2O2) and malondialdehyde (MDA) concentrations. The activity of the antioxidant enzymes, peroxidase (POD) and catalase (CAT), were found to be enhanced in response to increasing metal concentrations. Superoxide dismutase (SOD) activity decreased after Zn treatment but increased after Mo treatment. A marked increase in POD and CAT in response to metal stress suggests that these enzymes might have a significant cooperative role in regulating H2O2 production, although CAT, in response to drought and salt stress, has been reported to only play a supplementary role in A. orientalis. These results indicated that A. orientalis is susceptible to long-term Zn stress but can tolerate up to 2.5 mM Mo in the long-term. Deficiency of Mo is more common than high toxic concentrations in the environment. Therefore Zn contamination should be considered as one of the major threats for A. orientalis in its native habitat.

Neopterin-induced Suppression of Erythropoietin Production In Vitro

Pteridines ◽  
1995 ◽  
Vol 6 (1) ◽  
pp. 12-16 ◽  
Author(s):  
W. Schobersberger ◽  
W. Jelkmann ◽  
J. Fandrey ◽  
S. Frede ◽  
H. Wachter ◽  
...  
Keyword(s):  
Inhibitory Effect ◽  
Inhibitory Influence ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Anemia Of Chronic Disease ◽  
Erythropoietin Production ◽  
High Concentrations ◽  
Dose Dependent

Summary The production of neopterin increases in several diseases with activation of the ceIlular immune response. As previously shown serum concentrations of neopterin are inversely correlated with blood hemoglobin concentrations in the anemia of hematological and malignant disorders. Besides the role of chronic immune activation on the disturbed iron metabolism, an inhibitory influence of pteridines on cellular erythropoietin production could not be excluded. To test the possibility that pteridines are able to suppress the hypoxia-induced production of erythropoietin, the effects of neopterin and 7,8-dihydroneopterin on the human ceIl line HepG2 (hepatoceIlular carcinoma) were investigated. 24 h incubation with neopterin induced a dose-dependent reduction of erythropoietin production. The erythropoietin concentration significantly decreased by - 57.6% with 300 11M and by - 34.9% with 100 11M neopterin, respectively. 7,8 dihydroneopterin did not influence erythropoietin production. The inhibitory effect of neopterin on erythropoietin production was a consequence of reduced erythropoietin-mRNA levels. The results of this study show a neopterin-induced suppression of hypoxia-induced erythropoietin formation in HepG2 cultures in a dose dependent manner. We speculate that under in vivo conditions high concentrations of neopterin can aggravate the anemia of chronic disease.

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