Evaluation of Some Cytogenetic Effects of Hexane Extract from Tribulus terrestris in vivo and in vitro

The present study was carried out to evaluate the cytogenetic effects of the hexane extract of the plant Tribulus terrestris fruits. The cytogenetic evaluations involved mitotic index of bone marrow and spleen cells in albino male mice, micronucleus formation and sperm-head abnormalities. The micronucleus evaluations were further explored in blood lymphocyte cultures of healthy donors through an in vitro study. The search was carried out through three stages. In the first, the cytogenetic effects of three doses 5,10 and 20 mg/kg) of the plant extract were evaluated, while in the second stage, interactions Pre- and Post-treatments between the ideal dose 5 mg/kg of the plant extract and the drug Mitomycin-C (MMC). In stage three, the micronucleus formation was evaluated in the lymphocyte cultures of healthy individuals after treatment with three concentrations (5, 10 and 20 µg/ml) of the plant extract, in addition to interactions with the drug MMC which revealed significant mutagenic actions as judged by the investigated parameters. Reduced mitotic index, and increased frequencies of micronucleus formation (in vivo and in vitro) and sperm-head abnormalities were observed. The first stage revealed that the plant extract reduced the spontaneous formation of micronuclei and sperm-head abnormalities. The ideal dose 5 mg/kg of the plant extract was effective in modulating the mutagenic effects of the drug MMC. In this regard, the pre-treatment was more effective than post-treatment. The stage three showed that the plant extract was effective in reducing the spontaneous, as well as, MMC induced formation of micronuclei in lymphocyte cultures.

Download Full-text

Effects of chelating agents during freeze-drying of boar spermatozoa on DNA fragmentation and on developmental ability in vitro and in vivo after intracytoplasmic sperm head injection

Zygote ◽

10.1017/s0967199406003935 ◽

2007 ◽

Vol 15 (1) ◽

pp. 15-24 ◽

Cited By ~ 38

Author(s):

M. Nakai ◽

N. Kashiwazaki ◽

A. Takizawa ◽

N. Maedomari ◽

M. Ozawa ◽

...

Keyword(s):

Dna Fragmentation ◽

Freeze Drying ◽

Chelating Agents ◽

Sperm Head ◽

Control Group ◽

Sperm Dna Fragmentation ◽

Freeze Dried ◽

Boar Sperm

SUMMARYSuccessful offspring production after intracytoplasmic injection of freeze-dried sperm has been reported in laboratory animals but not in domesticated livestock, including pigs. The integrity of the DNA in the freeze-dried sperm is reported to affect embryogenesis. Release of endonucleases from the sperm is one of the causes of induction of sperm DNA fragmentation. We examined the effects of chelating agents, which inhibit the activation of such enzymes, on DNA fragmentation in freeze-dried sperm and on the in vitro and in vivo developmental ability of porcine oocytes following boar sperm head injection. Boar ejaculated sperm were sonicated, suspended in buffer supplemented with (1) 50 mM EGTA, (2) 50 mM EDTA, (3) 10 mM EDTA, or (4) no chelating agent and freeze-dried. A fertilization medium (Pig-FM) was used as a control. The rehydrated spermatozoa in each group were then incubated in Pig-FM at room temperature. The rate of DNA fragmentation in the control group, as assessed by the TUNEL method, increased gradually as time after rehydration elapsed (2.8% at 0 min to 12.2% at 180 min). However, the rates in all experimental groups (1–4) did not increase, even at 180 min (0.7–4.1%), which were all significantly lower (p < 0.05) than that of the control group. The rate of blastocyst formation after the injection in the control group (6.0%) was significantly lower (p < 0.05) than those in the 50 mM EGTA (23.1%) and 10 mM EDTA (22.6%) groups incubated for 120–180 min. The average number of blastocyst cells in the 50 mM EGTA group (33.1 cells) was significantly higher (p < 0.05) than that in the 10 mM EDTA group (17.8 cells). Finally, we transferred oocytes from 50 mM EGTA or control groups incubated for 0–60 min into estrous-synchronized recipients. The two recipients of the control oocytes became pregnant and one miscarried two fetuses on day 39.The results suggested that fragmentation of DNA in freeze-dried boar sperm is one of the causes of decreased in vitro developmental ability of injected oocytes to the blastocyst stage. Supplementation with EGTA in a freeze-drying buffer improves this ability.

Download Full-text

Changes in lipid diffusibility in the hamster sperm head plasma membrane during capacitation in vivo and in vitro

Molecular Reproduction and Development ◽

10.1002/(sici)1098-2795(199805)50:1<86::aid-mrd11>3.0.co;2-w ◽

1998 ◽

Vol 50 (1) ◽

pp. 86-92 ◽

Cited By ~ 15

Author(s):

T. Timothy Smith ◽

Christine A. Mckinnon-Thompson ◽

David E. Wolf

Keyword(s):

Plasma Membrane ◽

Sperm Head

Download Full-text

Characteristics and Biocompatibility of the Hydroxyapatite Based Two Ternary Biocomposites

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.720.130 ◽

2016 ◽

Vol 720 ◽

pp. 130-140

Author(s):

Berrak Bulut ◽

Ziya Engin Erkmen ◽

Eyup Sabri Kayali

Keyword(s):

Mechanical Properties ◽

Physical And Mechanical Properties ◽

Secondary Phase ◽

In Vitro Test ◽

Compression Tests ◽

Vitro Test ◽

Dental Applications ◽

The Ideal

Hydroxyapatite (HA) is a very popular bioceramic for orthopedic and dental applications. Although HA has excellent biocompatibility, its inferior mechanical properties make it unsuitable for load-bearing implant applications. Therefore, HA should be strengthened by a secondary phase for robust mechanical properties. The aim of this study was to compare the properties of HA-Al2O3 (HAC) and HA-ZrO2 (HZC) composites with the addition of 5 and 10 wt% commercial inert glass (CIG); independently. The mixture powders were pressed and then, the pellets were sintered between 1000-1300 °C for 4 hours. Microstructural characterizations were carried out using SEM + EDS and XRD, while hardness and compression tests were done to measure mechanical properties. In order to investigate the biocompatibility behavior of the samples in vitro and in vivo tests were performed. The mechanical properties of HAC composites increased with rising CIG content and increasing sintering temperature. For HZC composites, increasing CIG content caused an elevation in hardness and a decrease in compressive strength values at 1300 °C. The composites having the best physical and mechanical properties also showed improved bioactive properties at in vitro test. In this study, the ideal composite was selected as HZC5 sintered at 1200 °C depending on the microstructure, mechanical and biocompatibility properties.

Download Full-text

Delving into the Antiurolithiatic Potential of Tribulus terrestris Extract Through –In Vivo Efficacy and Preclinical Safety Investigations in Wistar Rats

Scientific Reports ◽

10.1038/s41598-019-52398-w ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Jyoti Kaushik ◽

Simran Tandon ◽

Rishi Bhardwaj ◽

Tanzeer Kaur ◽

Surinder Kumar Singla ◽

...

Keyword(s):

Body Weight ◽

Aqueous Extract ◽

Kidney Stones ◽

Wistar Rats ◽

Lethal Dose ◽

Tribulus Terrestris ◽

Oral Toxicity ◽

Preclinical Safety

Abstract Modern treatment interventions for kidney stones are wrought with side-effects, hence the need for alternative therapies such as plant-based medicines. We have previously documented through in vitro studies that statistically optimized aqueous extract of Tribulus terrestris (Zygophyllaceae family) possesses antiurolithic and antioxidant potential. This provides strong scientific foundation to conduct in vivo efficacy and preclinical safety studies to corroborate and lend further proof to its ability to prevent and cure kidney stones. The preventive and curative urolithiatic efficacy in experimentally induced nephrolithiatic Wistar rats, along with preclinical toxicity was evaluated following oral administration of statistically optimized aqueous extract of T. terrestris. Treatment showed augmented renal function, restoration of normal renal architecture and increase in body weight. Microscopic analysis of urine revealed excretion of small sized urinary crystals, demonstrating that treatment potentially modulated the morphology of renal stones. Tissue enzymatic estimation affirmed the antioxidant efficacy of treatment with reduced free radical generation. Significant upregulation of p38MAPK at both the gene and protein level was noted in hyperoxaluric group and interestingly treatment reversed it. Acute oral toxicity study established the Median Lethal Dose (LD50) to be greater than 2000 mg/kg body weight (b.wt.) No observed adverse effect level (NOAEL) by repeated oral toxicity for 28 days at 750 mg/kg b.wt. was noted. This study lends scientific evidence to the safe, preventive and curative potential of statistically optimized aqueous extract of T. terrestris at a dose of 750 mg/kg b.wt. and suggests that the extract shows promise as a therapeutic antiurolithic agent.

Download Full-text

ACUTE TOXICITY STUDY OF THE LEAVES ETHANOLIC EXTRACT OF PICRIA FEL-TERRAE LOUR.

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2018.v11s1.26567 ◽

2018 ◽

Vol 11 (13) ◽

pp. 55

Author(s):

Popi Patilaya ◽

Dadang Irfan Husori ◽

Imam Bagus Sumantri ◽

Simon Sihombing

Keyword(s):

Acute Toxicity ◽

Plant Extract ◽

Aspartate Aminotransferase ◽

Alanine Aminotransferase ◽

Ethanolic Extract ◽

Male Mice ◽

Plant Leaves ◽

Liver And Kidney

Objective: Picria fel-terrae belongs to family Linderniaceae is also known as Pugun tano by Indonesian people. The ethanolic extract of plant leaves has several potential pharmacological activities including antidiabetic, anthelmintic, and antioxidant. However, the toxicity of the plant extract is rarely explored. This work was to investigate toxicity of the leaf ethanolic extract of P. fel-terrae on Artemia salina and male mice.Methods: Acute toxicity of the plant extract was studied by in vitro and in vivo methods. In vitro study was carried out by exposing nauplii to the plant extract at concentrations of 10, 100, 200, 500, and 1000 μg/ml for 48 h. In vivo study was performed on male mice that divided into four groups. Groups I, II, III, and IV were treated with sodium carboxymethyl cellulose 0.5%, the ethanolic extract of plant leaves at doses of 1000, 2000, and 5000 mg/kg bw, respectively. The animal toxic symptoms were observed every day for 14 days. On day 15, the blood of mice was collected to measure alanine aminotransferase, aspartate aminotransferase, and creatinine levels. The effects of plant extract on vital animal organs such as heart, liver, and kidney were also studied. Statistical analysis of data was performed using analysis of variance and followed by Tukey post hoc.Results: The results showed that the leaf ethanolic extract of P. fel-terrae to have weakly toxicity on A. salina with the LC50 of 768.07 μg/ml. At in vivo studies, the toxic symptoms of mice were not identified during experiment with all doses of the plant extract for 14 days. In addition, aspartate aminotransferase and creatinine levels were no significantly different between control and all treatment groups (p>0.05). However, alanine aminotransferase level changed when mice were exposed by the plant extract at the doses of 2.000 and 5.000 mg/kg bw. Although the mice were not dead during experiment, the animal organs such as heart, liver, and kidney were histologically changed.Conclusion: This study suggests that the ethanolic extract of P. fel-terrae leaves has weakly toxicity on A. salina and causes histological changes on male mice organs at the high doses.

Download Full-text

In vivo and in vitro studies of the cytogenetic effects ofCannabis sativa in rats and men

Teratology ◽

10.1002/tera.1420090110 ◽

1974 ◽

Vol 9 (1) ◽

pp. 81-85 ◽

Cited By ~ 15

Author(s):

Patricia A. Martin ◽

Marigold J. Thorburn ◽

Sybil A. Bryant

Keyword(s):

In Vitro Studies ◽

Cytogenetic Effects

Download Full-text

Cytogenetic effects in peripheral blood lymphocytes in descendants of chernobyl disaster liquidators under the impact of mitomycin c in vitro and folic acid in vivo

Cytology and Genetics ◽

10.3103/s0095452713010039 ◽

2013 ◽

Vol 47 (1) ◽

pp. 53-57 ◽

Cited By ~ 2

Author(s):

V. I. Kovaleva ◽

N. V. Bagatskaya

Keyword(s):

Folic Acid ◽

Mitomycin C ◽

Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Blood Lymphocytes ◽

Chernobyl Disaster ◽

Cytogenetic Effects ◽

The Impact

Download Full-text

Defective sperm head decondensation undermines the success of ICSI in the bovine

Reproduction ◽

10.1530/rep-17-0270 ◽

2017 ◽

Vol 154 (3) ◽

pp. 307-318 ◽

Cited By ~ 8

Author(s):

Luis Águila ◽

Ricardo Felmer ◽

María Elena Arias ◽

Felipe Navarrete ◽

David Martin-Hidalgo ◽

...

Keyword(s):

Calcium Oscillations ◽

Sperm Head ◽

Oocyte Activation ◽

Bovine Sperm ◽

Bovine Oocytes ◽

Mouse Eggs ◽

Do So ◽

Nuclear Decondensation

The efficiency of intracytoplasmic sperm injection (ICSI) in the bovine is low compared to other species. It is unknown whether defective oocyte activation and/or sperm head decondensation limit the success of this technique in this species. To elucidate where the main obstacle lies, we used homologous and heterologous ICSI and parthenogenetic activation procedures. We also evaluated whetherin vitromaturation negatively impacted the early stages of activation after ICSI. Here we showed that injected bovine sperm are resistant to nuclear decondensation by bovine oocytes and this is only partly overcome by exogenous activation. Remarkably, when we used heterologous ICSI,in vivo-matured mouse eggs were capable of mounting calcium oscillations and displaying normal PN formation following injection of bovine sperm, althoughin vitro-matured mouse oocytes were unable to do so. Together, our data demonstrate that bovine sperm are especially resistant to nuclear decondensation byin vitro-matured oocytes and this deficiency cannot be simply overcome by exogenous activation protocols, even by inducing physiological calcium oscillations. Therefore, the inability of a suboptimal ooplasmic environment to induce sperm head decondensation limits the success of ICSI in the bovine. Studies aimed to improve the cytoplasmic milieu ofin vitro-matured oocytes and to replicate the molecular changes associated within vivocapacitation and acrosome reaction will deepen our understanding of the mechanism of fertilization and improve the success of ICSI in this species.

Download Full-text