scholarly journals Assessment of Cytotoxicity, Fetotoxicity, and Teratogenicity ofPlathymenia reticulataBenth Barks Aqueous Extract

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Lia de Barros Leite Albuquerque ◽  
Cháriston André Dal Belo ◽  
Marcio Galdino dos Santos ◽  
Patricia Santos Lopes ◽  
Marli Gerenutti ◽  
...  
Keyword(s):  
Aqueous Extract ◽  
Reproductive Toxicity ◽  
Late Pregnancy ◽  
Control Group ◽  
Corpora Lutea ◽  
Experimental Conditions ◽  
Pregnant Rats ◽  
Skeletal Analysis ◽  
Ld50 Test

Scientific assessment of harmful interactions of chemicals over the entire reproductive cycle are divided into three segments based on the period: from premating and mating to implantation (I), from implantation to major organogenesis (II), and late pregnancy and postnatal development (III). We combined the segments I and II to assessPlathymenia reticulataaqueous extract safety. In order to investigate reproductive toxicity (segment I), pregnant rats received orally 0.5 or 1.0 g/kg of extract, daily, during 18 days. These concentrations were determined by a preliminaryin vitroLD50 test in CHO-k1 cells. A control group received deionized water. The offspring was removed at the 19th day, by caesarean, and a teratology study (segment II) was carried out. The corpora lutea, implants, resorptions, live, and dead fetuses were then counted. Placenta and fetuses were weighted. External and visceral morphology were provided by the fixation of fetuses in Bouin, whereas skeletal analysis was carried out on the diaphanizated ones. The increase in the weights of placenta and fetuses was the only abnormality observed. Since there was no sign of alteration on reproduction parameters at our experimental conditions, we conclude thatP. reticulataaqueous extract is safe at 0.5 to 1.0 g/kg and is not considered teratogenic.

Effect of interferon-γ treatment on the expression of interleukin-1β at the maternal - fetal interface of pregnant rats

2007 ◽  
Vol 19 (3) ◽  
pp. 510 ◽  
Author(s):  
Hong-Fei Xia ◽  
Quan-Hong Sun ◽  
Jing-Pian Peng
Keyword(s):  
Late Pregnancy ◽  
Interferon Γ ◽  
Control Group ◽  
Cytokine Network ◽  
Pregnant Rats ◽  
Endometrial Stromal Cells ◽  
Protein Levels ◽  
Decidua Basalis ◽  
Ifn Γ

In the present study, the possible mechanisms by which interferon (IFN)-γ affects pregnancy were investigated using the cytokine network model. The IFN-γ-induced expression of interleukin (IL)-1β was examined using western blotting, immunohistochemistry and immunofluorescence. The results showed that IFN-γ treatment significantly decreased the expression of uterine IL-1β protein during the preimplantation, post-implantation and mid-gestation periods. The expression of IL-1β protein was increased after IFN-γ treatment compared with the control group in late pregnancy. In the placenta, IL-1β protein levels were significantly increased after IFN-γ treatment in early and mid-pregnancy. In late pregnancy, IFN-γ treatment significantly decreased placental IL-1β protein levels. IL-1β was mainly expressed in the myometrium, uterine arteries, decidua basalis, trophospongium of the junctional layer and trophoblastic epithelium of the labyrinthine layers. IL-1β was mainly located in the cytoplasm of in vitro cultured endometrial stromal cells (ESCs). IFN-γ treatment did not affect the distribution of IL-1β, only the expression of IL-1β. The effects of IFN-γ on the proliferation of ESCs were determined using an MTS (a novel tetrazolium compound) assay. IFN-γ treatment inhibited the proliferation of ESCs and decreased the weight of the fetus and placenta. These results indicate that exogenous IFN-γ affects the expression of IL-1β and inhibits ESC proliferation.

scholarly journals In vitro activity of zinc oxide-eugenol and glass ionomer cements on Candida albicans

2005 ◽  
Vol 19 (2) ◽  
pp. 134-138 ◽  
Author(s):  
Anna Carolina Aguiar Cassanho ◽  
Aletéia Massula Fernandes ◽  
Luciane Dias de Oliveira ◽  
Claudio Antonio Talge Carvalho ◽  
Antonio Olavo Cardoso Jorge ◽  
...  
Keyword(s):  
Zinc Oxide ◽  
Candida Albicans ◽  
Control Group ◽  
Glass Ionomer ◽  
Experimental Conditions ◽  
Mean Values ◽  
Zinc Oxide Eugenol ◽  
Colony Forming Units ◽  
And Control

The aim of this study was to evaluate in vitro the antimicrobial activity of glass ionomer (GIC) and zinc oxide-eugenol (ZOE) cements against Candida albicans. Standardized GIC and ZOE specimens were maintained in contact with C. albicans suspension (1 <FONT FACE=Symbol>´</FONT> 10(6) cells/ml) at 37°C for 24 h, 48 h or 7 days. A control group without any testing cement was included. After the incubation period, aliquots of 0.1 ml were plated on Sabouraud's agar, and then the number of colonies was counted. The results were expressed as values of logarithms of colony-forming units per milliliter (log CFU/mL) and were analyzed statistically by Kruskal-Wallis ANOVA. After 48 h of incubation, the ZOE group presented no growth of C. albicans. GIC and control groups presented similar mean values at all tested periods. According to the results obtained, it could be concluded that, under the experimental conditions, ZOE cement was more effective in vitro against C. albicans than GIC.

scholarly journals Silica nanoparticles inducing the apoptosis of spermatocyte cell through microRNA-450b-3p targeting MTCH2-mediating mitochondrial signaling pathway

2020 ◽  
Author(s):  
Guiqing Zhou ◽  
Jianhui Liu ◽  
Xiangyang Li ◽  
Yujian Sang ◽  
Yue Zhang ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Silica Nanoparticles ◽  
Caspase 3 ◽  
Reproductive Toxicity ◽  
Control Group ◽  
Caspase 9 ◽  
Protein Expressions ◽  
Underlying Mechanisms

Abstract Background: Silica nanoparticles (SiNPs) are found in environmental particulate matter and are proven to have adverse effects on fertility. The relationship and underlying mechanisms between miRNAs and apoptosis induced by SiNPs during spermatogenesis is currently ambiguous. Experimental design: The present study was designed to investigate the role of miRNA-450b-3p in the reproductive toxicity caused by SiNPs. In vivo, 40 male mice were randomly divided into control and SiNPs groups, 20 per group. The mice in the SiNPs group were administrated 20 mg/kg SiNPs by tracheal perfusion once every 5 days, for 35 days, and the control group were given the equivalent of a normal luminal saline. In vitro, spermatocyte cells were divided into 0 and 5 μg/mL SiNPs groups, after passaged for 30 generations, the GC-2spd cells in 5 μg/mL SiNPs groups were transfected with miRNA-450b-3p and its mimic and inhibitor. Results: In vivo, the results showed that SiNPs damaged tissue structures of testis, decreased the quantity and quality of the sperm, reduced the expression of miR-450b-3p, and increased the protein expressions of the MTCH2, BID, BAX, Cytochrome C, Caspase-9, and Caspase-3 in the testis. In vitro, SiNPs obviously repressed the viability and increased the LDH level and apoptosis rate, decreased the levels of the miR-450b-3p, significantly enhanced the protein expressions of the MTCH2, BID, BAX, Cytochrome C, Caspase-9, Caspase-3; while the mimic of miR-450b-3p reversed the changes induced by SiNPs, but inhibitor further promoted the effects induced by SiNPs.Conclusion: The result suggested that SiNPs could induce the spermatocyte apoptosis by inhibiting the miR-450b-3p expression to target promoting the MTCH2 resulting in activating mitochondrial apoptotic signaling pathways in the spermatocyte cells.

335 CYTOPLASMIC MICROINJECTION OF EXOGENOUS DNA IN IN VITRO AND IN VIVO DERIVED SHEEP EMBRYOS

2011 ◽  
Vol 23 (1) ◽  
pp. 263
Author(s):  
F. Pereyra-Bonnet ◽  
A. Gibbons ◽  
M. Cueto ◽  
R. Bevacqua ◽  
L. Escobar ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Genetically Modified ◽  
Fold Increase ◽  
Egfp Expression ◽  
Control Group ◽  
Corpora Lutea ◽  
Exogenous Dna ◽  
Frozen Semen

Microinjection of DNA into the male pronucleus is a commonly used method to generate transgenic animals. However, it is only moderately efficient in several species because it requires proper male pronuclear visualisation, which occurs only in a narrow window of time in mice. The cytoplasmic microinjection of exogenous DNA (eDNA) is an alternative method that has not been fully investigated. Our objective was to evaluate if cytoplasmic microinjection of eDNA is capable of producing genetically modified embryos. In vitro and in vivo derived sheep embryos were cytoplasmically microinjected with pCX-EGFP previously incubated (5 min in a PVP droplet) with oolemma-cytoplasm fragments obtained from donor oocytes by microsurgery. A control group using microinjected plasmid alone was included in the in vivo procedure. For in vitro microinjection, IVF embryos were microinjected with circular plasmid with promoter (50 or 500 ng μL–1) or without promoter (50 ng μL–1) at 6 h after fertilization. The IVF was performed following (Brackett and Olliphant 1975 Biol. Reprod. 12, 260–274) with 15 × 106 spermatozoa mL–1, and presumptive zygotes were cultured in SOF. The expression of enhance green fluorescent protein (EGFP) was determined under blue light. For in vivo microinjection, embryos from superovulated sheep (by standard procedures) were recovered and microinjected with 50 ng μL–1 of linearized plasmid without promoter at 12 h after laparoscopic insemination with frozen semen (100 × 106 spermatozoa per sheep). Plasmid without promoter was used to avoid any possible cytotoxic effect produced by EGFP expression. The microinjection of IVF embryos with 50 ng μL–1 of plasmid was the best condition to produce embryos expressing eDNA (n = 96; 46.9% cleaved; 12.2% blastocysts; 53.0 and 4.1% of green embryos and blastocysts, respectively). Variables between the groups with or without promoter IVF were not statistically different (Fisher test: P < 0.05); however, when 500 ng μL–1 was microinjected, no blastocysts were obtained. In the in vivo embryo production group, 111 presumptive zygotes were microinjected (n = 37; with plasmid alone) from 16 donor sheep (11.5 ± 4.0 corpora lutea; 8.4 ± 4.8 presumptive zygotes recovered; 74.3% recovery rate). The mean time from injection to cleavage was 18.0 ± 4.5 h, and the percentage of cleavage and damage (due to the embryo injection) were >70% and <10%, respectively. Fifty-eight good quality embryos were transferred into the oviducts of 19 surrogate ewes; 12 of them are pregnant (63.1%). The presence of green IVF embryos demonstrates that eDNA was transported to the nucleus after cytoplasmic injection. We believe that the multi-fold increase (50- to 100-fold) in plasmid concentration compared with that used by others was the key step to our successful cytoplasmic microinjection. Accordingly, the new/old methodology described in this study provides an easy DNA construct delivery system of interest for the implementation of early reprogramming events. In addition, results obtained in the near future using in vivo cytoplasmic microinjection with high concentrations of eDNA could revalidate this technique for producing genetically modified large animals.

scholarly journals Hormonal control of Luteal 20α-Hydroxy steroid dehydrogenase and Δ5-3β-hydroxy steroid dehydrogenase during luteolysis in the pregnant rat

1975 ◽  
Vol 152 (3) ◽  
pp. 433-443 ◽  
Author(s):  
R G Rodway ◽  
N J Kuhn
Keyword(s):  
Prostaglandin F2α ◽  
Late Pregnancy ◽  
Normal Pregnancy ◽  
Hormonal Control ◽  
Placental Lactogen ◽  
Pregnant Rats ◽  
Pregnant Rat ◽  
Hydroxy Steroid ◽  
Steroid Dehydrogenase

Treatment of pregnant rats with human chorionic gonadotrophin, luteotrophin (luteinizing hormone), luteotrophin-releasing hormone, prostaglandin F2α, aminoglutethimide, or by foetoplacental removal or hysterectomy achieved a common multiple-response pattern, namely increased activity of luteal 20α-hydroxy steroid dehydrogenase with decreased activity of delta5-3β-hydroxy steriod dehydrogenase and release of delta4-3-oxo steroids in vitro. 2. Similar effects of foetoplacental removal are noted in pregnant mice. 3. Gonadotrophin induced lower activities of 20α-hydroxy steroid dehydrogenase, except at the very end of pregnancy, and partly inhibited the induction caused by foetoplacental removal. 4. The results suggest that existence of a placental factor that restrains these changes until the end of normal pregnancy, which is produced in amounts proportional to the number of placentae and is conveyed to the ovary via the blood. 5. This factor was not replaced by prolactin. 6. It is argued that neither placental lactogen nor pituitary luteotrophin participate in the induction of 20α-hydroxy steroid dehydrogenase at late pregnancy in the rat. 7. Aminoglutethimide induced 20α-hydroxy steroid dehydrogenase only in late pregnancy. This was partly reversed by progesterone, wholly reversed by progesterone plus oestrogen, and did not involve the pituitary.

Pregnancy Does Not Increase Susceptibility to Bupivacaine in Spinal Root Axons

1997 ◽  
Vol 87 (3) ◽  
pp. 610-616 ◽  
Author(s):  
Friederike B. Dietz ◽  
Richard A. Jaffe
Keyword(s):  
Conduction Block ◽  
Late Pregnancy ◽  
Underlying Mechanism ◽  
Ventral Root ◽  
Spinal Root ◽  
Pregnant Rats ◽  
C Fibers ◽  
Antinociceptive Effects ◽  
The Difference

Background The underlying mechanism of enhanced antinociceptive effects and increased susceptibility to local anesthetics during pregnancy is not known. Mechanical, hormonal, biochemical, and neural changes have been suggested. The authors measured the susceptibility of individual spinal root axons to bupivacaine during late pregnancy in rats and compared them with similar measurements in nonpregnant rats. Methods Lumbar dorsal and ventral roots were excised from anesthetized pregnant and nonpregnant rats. Single-fiber dissection and recording techniques were used to isolate activity in individual axons. Supramaximal constant voltage stimuli were delivered to the distal end of the root. During in vitro perfusion, each root was exposed to increasing concentrations of bupivacaine, and the minimum blocking concentration (Cm) and the concentration that increased conduction latency by 50% (EC50) were measured. Results Myelinated and unmyelinated dorsal and ventral root axons of pregnant rats appeared to be less sensitive to steady-state conduction block and to the latency-increasing effects of bupivacaine than were equivalent axons from nonpregnant rats. Although when comparing specific axon types, only the difference in C-fibers was significant (Cm = 29.8 microM for pregnant and Cm = 22.1 microM for nonpregnant rats, P &lt; 0.05; EC50 = 19.9 microM and 13.6 microM, respectively). Conclusions In contrast to clinical expectations, the susceptibility to bupivacaine conduction block in individual dorsal and ventral root axons during late pregnancy in rats was not greater in pregnant animals. Pregnancy-related changes in diffusion barriers and activation of endogenous analgesic systems without changes in the electrophysiologic properties of spinal root axons are suggested as possible explanations for the discrepancy between clinical and experimental observations.

scholarly journals Heat-shock protein-25/27 phosphorylation by the δ isoform of protein kinase C

1998 ◽  
Vol 332 (3) ◽  
pp. 703-712 ◽  
Author(s):  
Evelyn T. MAIZELS ◽  
Carl A. PETERS ◽  
Michael KLINE ◽  
Richard E. CUTLER ◽  
Malathy SHANMUGAM ◽  
...  
Keyword(s):  
Heat Shock ◽  
Protein Kinase ◽  
Late Pregnancy ◽  
Mitogen Activated Protein Kinase ◽  
Corpora Lutea ◽  
Hsp 27 ◽  
Pkc Isoforms ◽  
Mitogen Activated Protein

Small heat-shock proteins (sHSPs) are widely expressed 25–28 kDa proteins whose functions are dynamically regulated by phosphorylation. While recent efforts have clearly delineated a stress-responsive p38 mitogen-activated protein-kinase (MAPK)-dependent kinase pathway culminating in activation of the heat-shock (HSP)-kinases, mitogen-activated protein-kinase-activated protein kinase-2 and -3, not all sHSP phosphorylation events can be explained by the p38 MAPK-dependent pathway. The contribution of protein kinase C (PKC) to sHSP phosphorylation was suggested by early studies but later questioned on the basis of the reported poor ability of purified PKC to phosphorylate sHSP in vitro. The current study re-evaluates the role of PKC in sHSP phosphorylation in the light of the isoform complexity of the PKC family. We evaluated the sHSP phosphorylation status in rat corpora lutea obtained from two stages of pregnancy, mid-pregnancy and late-pregnancy, which express different levels of the novel PKC isoform, PKC-δ. Two-dimensional Western blot analysis showed that HSP-27 was more highly phosphorylated in vivo in corpora lutea of late pregnancy, corresponding to the developmental stage in which PKC-δ is abundant and active. Late-pregnant luteal extracts contained a lipid-sensitive HSP-kinase activity which exactly co-purified with PKC-δ using hydroxyapatite and S-Sepharose column chromatography. To determine whether there might be preferential phosphorylation of sHSP by a particular PKC isoform, purified recombinant PKC isoforms corresponding to those PKC isoforms detected in rat corpora lutea were evaluated for HSP-kinase activity in vitro. Recombinant PKC-δ effectively catalysed the phosphorylation of sHSP in vitro, and PKC-α was 30–50% as effective as an HSP-kinase; other PKCs tested (β1, β2, ε and ζ) were poor HSP-kinases. These results show that select PKC family members can function as direct HSP-kinases in vitro. Moreover, the observation of enhanced luteal HSP-27 phosphorylation in vivo, in late pregnancy, when PKC-δ is abundant and active, suggests that select PKC family members contribute to sHSP phosphorylation events in vivo.

scholarly journals Investigation of ginger (Zingiber officinale) aqueous extract as an anti-diabetic in vitro

2021 ◽  
Vol 913 (1) ◽  
pp. 012108
Author(s):  
P Pakan ◽  
K Lidia ◽  
M Riwu
Keyword(s):  
Side Effects ◽  
Glucose Uptake ◽  
Aqueous Extract ◽  
Zingiber Officinale ◽  
Control Group ◽  
Plain Water ◽  
Ginger Extract ◽  
Mouse Tissues ◽  
Treatment Of Diabetes

Abstract Diabetes mellitus is a condition of metabolic imbalance, indicated by a high level of blood glucose (hyperglycemia) resulting from a reduction of insulin secretion, action, or both. People with diabetes suffer from a lack or deficiency of insulin or insulin resistance. The metabolic imbalances are often not satisfactorily corrected using conventional medicines and even cause some side effects, which can be detrimental. Research on herbal medicines for the treatment of diabetes is urged by the need to reduce unwanted side effects common with conventional medicines/treatments used in glucose regulation. This study aims to investigate the antidiabetic effect of Ginger (Zingiber officinale) aqueous extract in improving the glucose uptake in mouse tissues in vitro. This study is a true experimental research design with a posttest-only control group design. There were three groups of mice in this study: the control group, which were only given plain water; the second group of mice with 5% aqueous ginger extract and the last group were given 25% aqueous ginger extract. All groups were given treatment for four consecutive weeks, then dissected their cardiac muscle, skeletal muscle, pancreas, and liver tissues to analyze the glucose uptake. The result showed that both the ginger aqueous extract groups were able to increase the glucose uptake of the mice. In conclusion, this research has shown that aqueous ginger extract may have improved the glucose uptake in most tissues of the mice in the groups. Therefore, ginger could have great potential as an alternative way in the treatment of diabetes type 2.

Localization and regulation of IL-1α in rat myometrium during late pregnancy and the postpartum period

2001 ◽  
Vol 280 (3) ◽  
pp. R879-R888 ◽  
Author(s):  
J. Andres Melendez ◽  
James M. Vinci ◽  
John J. Jeffrey ◽  
Brian D. Wilcox
Keyword(s):  
Postpartum Period ◽  
Preterm Labor ◽  
Interleukin 1 ◽  
Late Pregnancy ◽  
Mrna Levels ◽  
Pregnant Rats ◽  
Low Levels ◽  
Interstitial Collagenase

Interleukin-1 (IL-1) has been implicated as a participant in preterm labor that is induced by bacterial infection. Previously, we showed that serotonin-induced production of IL-1α by myometrial smooth muscle cells in vitro is also essential for the synthesis of interstitial collagenase. It is therefore likely that IL-1α production in uterine tissues has implications for both the normal physiology of involution and for the pathophysiological mechanisms of preterm labor. The objective of this study was to characterize the serotonin-induced production of IL-1α by myometrial cultures in vitro and to assess the production of IL-1α and its relationship to collagenase production in vivo during pregnancy and the postpartum period. Immunohistochemistry demonstrated IL-1α protein in the nuclei and cytoplasm of serotonin-treated myometrial cells. IL-1α levels were decreased by treatment with progesterone or IL-1-receptor antagonist but were unaffected by lipopolysaccharide. Western analysis of myometrium from pregnant rats showed low levels of IL-1α during midpregnancy with increased concentrations at days 21 and 22 and postpartum. IL-1α mRNA levels also increased from days 15to 22. Levels of mRNA for IL-1β also increased, although to a lesser degree than IL-1α. Both mRNAs decreased postpartum. Conversely, mRNA for interstitial collagenase was barely detectable at term but increased postpartum. Together, these data show that serotonin stimulates IL-1α production in vitro and indicate that normal myometrium from pregnant rats is an identifiable source of IL-1 during late pregnancy. The findings are consistent with the possibility that myometrial IL-1α participates in normal labor as well as the postpartum production of interstitial collagenase.

scholarly journals Effect of insulin on glucose metabolism in adipocytes from virgin and late-pregnant rats

1984 ◽  
Vol 224 (2) ◽  
pp. 685-688 ◽  
Author(s):  
A Leturque ◽  
M Guerre-Millo ◽  
M Lavau ◽  
J Girard
Keyword(s):  
Glucose Metabolism ◽  
High Sensitivity ◽  
Late Pregnancy ◽  
Maximal Response ◽  
Maximal Effect ◽  
Insulin Stimulation ◽  
Response Curves ◽  
Pregnant Rats

Under basal conditions (zero insulin), paraovarian adipocytes from 19-day-pregnant rats exhibited the same rates of [U-14C]glucose conversion into CO2 and total lipids as did those from age-matched virgin rats. The dose-response curves for insulin stimulation of glucose metabolism were similar in both groups: maximal response (+100% over basal values) and high sensitivity (half-maximal effect at 0.05 nM-insulin). The present results suggest that the insulin resistance in vivo that occurs during late pregnancy may involve circulating factors lost in vitro.

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