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1.
Stem Cells Int ; 2022: 9346767, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35371264

RESUMO

Pluripotent stem cells (PSCs) are considered as a potent tool for use in regenerative medicine. Highly efficient generation of PSCs through chromatin modulators such as trichostatin A (TSA) might change their MHC molecule expression profile. The efficiency of PSC generation and their immunogenicity is major obstacles for clinical use. Hence, we aim to investigate whether the use of TSA during PSC generation affects MHC expression level. Three PSC lines were generated by iPSCs, NT-ESCs, and IVF-ESCs' reprogramming methods from B6D2F1 mouse embryonic fibroblast cells. Established PSC lines were characterized by alkaline phosphatase assay (ALP) and immunocytochemistry. Their chromosome fidelity was checked by karyotyping. The expression level of pluripotent genes (oct4, nanog, sox2, klf4), HDACs (hdac1, hdac2, and hdac3), and immune-related genes (including Qa-1, Qa-2, H2kb, H2kd, H2db, H2db, CIITA, H2-IE-ßb, H2-IE-ßd) in iPSC and ESC lines were assessed by real-time PCR analysis. The presence of MHC molecules on the surface of pluripotent stem cells was also checked by flow cytometry technique. Significant increase of pluripotency markers, oct4, nanog, sox2, and klf4, was observed in 100 nM TSA-treated samples. 100 nM TSA induced significant upregulation of H2db in generated iPSCs. H2-IE-ßd was remarkably downregulated in 50 and 100 nM TSA-treated iPSC lines. The expression level of other immune-related genes was not greatly affected by TSA in iPSC and NT-ESC lines. It is concluded that the use of short-term and low concentration of TSA during reprogramming in PSC generation procedure significantly increases PSC generation efficiency, but do not affect the MHC expression in established cell lines, which is in the benefit of cell transplantation in regenerative medicine.

2.
Sci Rep ; 12(1): 9655, 2022 06 10.
Artigo em Inglês | MEDLINE | ID: mdl-35688923

RESUMO

Nowadays, the presence of excessive ions in water resources is of utmost concern and has attracted increasing attention; therefore, excessive amounts of these ions such as fluoride should be removed from drinking water. Conventional water treatment processes are shown to be incapable of the complete removal of redundant fluoride from aqueous water bodies, whereas adsorption is a promising, effective, cost-benefit, and simple method for this purpose. This study aimed to synthesize effective adsorbents from bivalve shells and evaluate the adsorption function of bivalve shells in removing fluoride from aqueous solutions. In this study, the oyster shell was collected from the Persian Gulf's seaside and were crushed by manual mortar and blender, and graded with standard sieves with 70 mesh size. The prepared bivalve shell was characterized by SEM and FTIR. To investigate and optimize various variables on fluoride removal percentage a response surface methodology based on central composite design (RSM-CCD) was used. Under optimal conditions (pH: 5.5, adsorbent dose: 0.3 g/L, contact time: 85 min and fluoride concentration: 3 mg/L) the maximum removal efficiency was 97.26%. Results showed that the adsorption equilibrium and kinetic data were matched with the isotherm Langmuir Model (R2 = 0.98) with qmax = 27.31 mg/g and pseudo-second-order reaction (R2 = 0.99). Also, a thermodynamic study exhibited that the adsorption process of fluoride into bivalve shells was an exothermic reaction and could not be a spontaneous adsorption process. Based on the results, the bivalve shell was found as an appropriate adsorbent to remove fluoride from aqueous solutions.


Assuntos
Poluentes Químicos da Água , Purificação da Água , Adsorção , Exoesqueleto/química , Animais , Fluoretos , Concentração de Íons de Hidrogênio , Cinética , Termodinâmica , Poluentes Químicos da Água/análise , Purificação da Água/métodos
3.
Eur Heart J Cardiovasc Imaging ; 23(7): 970-978, 2022 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-35511038

RESUMO

AIMS: Arrhythmogenic right ventricular cardiomyopathy (ARVC) is characterized by progressive fibro-fatty infiltration of the myocardium and associated with adverse cardiovascular (CV) events. This study aims to examine right atrial (RA) deformation in ARVC and understand its association with CV outcomes. METHODS AND RESULTS: RA strain was determined in 50 patients with definite ARVC, compared with a matched control group of 50 healthy individuals, and analysed for outcome association over a median follow-up duration of 5 years. A subgroup of 30 ARVC patients with normal RA volume (ARVC-N group) was compared with 30 matched controls (Control-N), and the outcome was analysed separately. RA reservoir, conduit, and pump strain were significantly impaired in ARVC vs. control. Similar observations were made in the N-ARVC subgroup. Reservoir strain was associated with an increased risk of atrial arrhythmia (AA) [hazard ratio (HR) 0.88, P < 0.01] and CV events (HR 0.92, P < 0.01). Conduit strain also predicted AA (HR 1.02, P < 0.01), while pump strain predicted CV events (HR 1.09, P = 0.02). Reservoir strain improved the fitness of bivariable models for the association of RV end-diastolic area index, RV fractional area change, and RV global longitudinal strain with CV events. CONCLUSION: ARVC patients display impaired RA strain even when RA volume is normal. Reservoir and pump strain are associated with an increased risk of CV events. Reservoir strain improved model fitness for the association of RVGLS and other echocardiographic parameters with CV events.


Assuntos
Displasia Arritmogênica Ventricular Direita , Apêndice Atrial , Ecocardiografia , Átrios do Coração/diagnóstico por imagem , Humanos , Miocárdio
4.
Heart ; 108(3): 225-232, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-33972358

RESUMO

OBJECTIVE: Left ventricular (LV) twist is a major component of ventricular mechanics reflecting the helical orientation of cardiac fibres and compensating for afterload mismatch. However, it is not known whether it determines outcome after transcatheter aortic valve implantation (TAVI). This study sought to investigate TAVI-induced short-term changes of LV twist and to define its role in outcome prediction. METHODS: A total of 146 patients (median age 81.78 years, 50.7% male) undergoing TAVI for severe aortic stenosis were included. LV rotation and twist were determined by speckle tracking echocardiography within 3 months before and 2 weeks after TAVI. All-cause mortality at 2 years was defined as primary end point. RESULTS: Patients who survived exhibited a higher apical peak systolic rotation (APSR) (p<0.001), twist (p=0.003) and torsion (p=0.019) pre-TAVI compared with those who died (n=22). Within 2 weeks after TAVI, APSR, twist and torsion decreased in patients who survived (all p<0.001), while no change occurred in those who died. Cox regression analysis showed an association of pre-TAVI APSR (HR 0.92, p=0.010), twist (HR 0.93, p=0.018) and torsion (HR 0.68, p=0.040) with all-cause mortality and an even stronger association of the respective changes after TAVI (∆APSR: HR 1.15, p<0.001; ∆twist: HR 1.14, p<0.001; ∆torsion: HR 2.53, p<0.001). All the parameters determined outcome independently of global longitudinal strain (GLS) and LV ejection fraction (LVEF). CONCLUSION: APSR, twist and torsion pre-TAVI as well as their change within 2 weeks after TAVI predict 2-year all-cause mortality after TAVI, adding incremental prognostic value to LVEF and GLS.


Assuntos
Estenose da Valva Aórtica , Substituição da Valva Aórtica Transcateter , Disfunção Ventricular Esquerda , Idoso de 80 Anos ou mais , Estenose da Valva Aórtica/complicações , Estenose da Valva Aórtica/diagnóstico por imagem , Estenose da Valva Aórtica/cirurgia , Feminino , Ventrículos do Coração/diagnóstico por imagem , Humanos , Masculino , Volume Sistólico , Substituição da Valva Aórtica Transcateter/efeitos adversos , Resultado do Tratamento , Disfunção Ventricular Esquerda/diagnóstico por imagem , Disfunção Ventricular Esquerda/etiologia , Função Ventricular Esquerda
5.
Zygote ; : 1-7, 2021 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-34823620

RESUMO

It has been documented that the inefficacy of round spermatid injection (ROSI) might be caused by abnormal epigenetic modifications. Therefore, this study aimed to evaluate the effect of trichostatin A (TSA) as an epigenetic modifier of preimplantation embryo development in activated ROSI oocytes. Matured oocytes were collected from superovulated female mice. Testes were placed in human tubal fluid medium and masses were then cut into small pieces to disperse spermatogenic cells. Round spermatids were treated with TSA and subsequently injected into oocytes. The expression level of the development-related genes including Oct4, Sox2, Nanog, Dnmt and Hdac transcripts were evaluated using qRT-PCR. Immunohistochemistry was performed to confirm the presence of Oct-4 protein at the blastocyst stage. There was no statistically significant difference in fertilization rate following ROSI/+TSA compared with the non-treated ROSI and intracytoplasmic sperm injection (ICSI) groups. Importantly, TSA treatment increased blastocyst formation from 38% in non-treated ROSI to 68%. The relative expression level of developmentally related genes increased and Dnmt transcripts decreased in ROSI/+TSA-derived embryos, similar to the expression levels observed in the ICSI-derived embryos. In conclusion, our results indicate that spermatid treatment with TSA prior to ROSI would increase the success rate of development to the blastocyst stage and proportion of pluripotent cells.

6.
Cardiovasc Ultrasound ; 19(1): 37, 2021 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-34802441

RESUMO

BACKGROUND: The effect of right ventricular (RV) leads on tricuspid valve has been already raised concerns, especially in terms of prognostic implication. For such assessment, three-dimensional transthoracic echocardiography (3D-TTE) has been used previously but there was no data on the use of post-procedural fluoroscopy in the literature. METHODS: We prospectively enrolled 59 patients who underwent clinically indicated placement of pacemaker or implantable cardioverter defibrillator (ICD). Vena contracta (VC) and tricuspid regurgitation (TR) severity were measured using two-dimensional transthoracic echocardiography (2D-TTE) at baseline. Follow up 3D-TTE was performed 6 months after device implantation to assess TR severity and RV lead location. RESULTS: Lead placement position in TV was defined in 51 cases.TR VC was increased after the lead placement, compared to the baseline study (VC: 3.86 ± 2.32 vs 3.18 ± 2.39; p = 0.005), with one grade worsening in TR in 25.4% of cases. The mean changes in VC levels were 1.14 ± 0.67 mm. Among all investigated parameters, VC changes were predicted based on lead placement position only in 3D-TTE (p < 0.001) while the other variables including fluoroscopy parameters were not informative. CONCLUSION: The RV Lead location examined by 3D-TTE seems to be a valuable parameter to predict the changes in the severity of the tricuspid regurgitation. Fluoroscopy findings did not improve the predictive performance, at least in short term follow up.


Assuntos
Ecocardiografia Tridimensional , Insuficiência da Valva Tricúspide , Ecocardiografia , Fluoroscopia , Humanos , Valva Tricúspide/diagnóstico por imagem , Insuficiência da Valva Tricúspide/diagnóstico
7.
Cell J ; 23(1): 109-118, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33650827

RESUMO

OBJECTIVE: In vitro maturation (IVM) of human oocytes is used to induce meiosis progression in immature retrieved oocytes. Calcium (Ca2+) has a central role in oocyte physiology. Passage through meiosis phase to another phase is controlled by increasing intracellular Ca2+. Therefore, the current research was conducted to evaluate the role of calcium ionophore (CI) on human oocyte IVM. MATERIALS AND METHODS: In this clinical trial study, immature human oocytes were obtained from 216 intracytoplasmic sperm injection (ICSI) cycles. After ovarian stimulation, germinal vesicle (GV) stage oocytes were collected and categorized into two groups: with and without 10 µM CI treatment. Next, oocyte nuclear maturation was assessed after 24-28 hours of culture. Real-time reverse transcription polymerase chain reaction (RT-PCR) was used to assess the transcript profile of several oocyte maturation-related genes (MAPK3, CCNB1, CDK1, and cyclin D1 [CCND1]) and apoptotic-related genes (BCL-2, BAX, and Caspase-3). Oocyte glutathione (GSH) and reactive oxygen species (ROS) levels were assessed using Cell Tracker Blue and 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) fluorescent dye staining. Oocyte spindle configuration and chromosome alignment were analysed by immunocytochemistry. RESULTS: The metaphase II (MII) oocyte rate was higher in CI-treated oocytes (73.53%) compared to the control (67.43%) group, but this difference was not statistically significant (P=0.13). The mRNA expression profile of oocyte maturation-related genes (MAPK3, CCNB1, CDK1, and CCND1) (P<0.05) and the anti-apoptotic BCL-2 gene was remarkably up-regulated after treatment with CI (P=0.001). The pro-apoptotic BAX and Caspase-3 relative expression levels did not change significantly. The CI-treated oocyte cytoplasm had significantly higher GSH and lower ROS (P<0.05). There was no statistically significant difference in meiotic spindle assembly and chromosome alignment between CI treatment and the control group oocytes. CONCLUSION: The finding of the current study supports the role of CI in meiosis resumption of human oocytes. (Registration Number: IRCT20140707018381N4).

8.
Adv Pharmacol Pharm Sci ; 2021: 5018092, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34993484

RESUMO

OBJECTIVES: Diabetic cardiomyopathy (DC) has become one of the serious complications in diabetic cases. In this study, we aimed to explore the syringic acid (SYR) protective effect against diabetes-induced cardiac injury in experimental rats. METHODS: Rats were divided in control and streptozotocin-induced diabetic rats which were subdivided into diabetic controls, and three test groups (SYR at 25, 50, and 100 mg/kg) and the nondiabetic group received 100 mg/kg of SYR. All treatments were given SYR for 6 weeks. SYR effects on cardiac diagnostic markers, heart lipid peroxidation, protein carbonylation, antioxidant system, and changes of the heart mitochondrial mass and biogenesis were measured. RESULTS: Diabetes induction prompted CK-MB, LDH levels in serum, cardiac catalase, and superoxide dismutase activity, as well as cardiac TBARs and carbonylated protein. SYR administration (100 m/kg) attenuated CK-MB and LDH levels. Also, 50 and 100 mg/kg of SYR reduced cardiac TBARs and carbonylated protein in diabetic rats. These treatments did not show any effects on GSH content, mtDNA, and mitochondrial biogenesis indices (PGC1- α, NRF1, NRF2, and TFAM) in heart tissue. CONCLUSIONS: SYR treatment showed protective effects on diabetic cardiomyopathy in rats by reducing lipid peroxidation and protein carbonylation. The possible mechanisms could be related to antioxidant activity of this phenolic acid. SYR might play a role of a protective factor in cardiac challenges in diabetes.

9.
Int J Fertil Steril ; 14(2): 110-115, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32681622

RESUMO

BACKGROUND: Embryo vitrification is a key instrument in assisted reproductive technologies (ARTs). However, there is increasing concern that vitrification adversely affects embryo development. This study intends to assess the effect of vitrification on developmental competence, in addition to expressions of long non-coding RNA (lncRNA) gene trap locus 2 (Gtl2) and its reciprocal imprinted gene delta-like homolog 1 (Dlk1), in mouse blastocysts. MATERIALS AND METHODS: In this experimental study, we have designed three experimental groups: control (fresh blastocysts collected from superovulated mice), in vitro fertilization (IVF; blastocysts derived from IVF) and vitrification (IVF derived blastocysts subjected to vitrification/warming at the 2-cell stage). Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to assess the expression levels of Gtl2 and Dlk1 in the blastocysts. RESULTS: The results showed that vitrification group had significantly lower blastocyst and hatching rates compared to the IVF group (P<0.037) and (P<0.041), respectively. Gtl2 was down-regulated and Dlk1 was up-regulated following the IVF and vitrification (P<0.05). CONCLUSION: These results suggested that IVF and vitrification disturbed genomic imprinting and lncRNA gene expressions, which might affect the health of IVF children.

10.
J Cell Physiol ; 235(12): 9752-9762, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32415675

RESUMO

Despite encouraging advances in fertility technology, the success rate of an ongoing pregnancy is relatively low and predominantly associated with implantation failure. Inflammatory responses are beneficial in the fetomaternal interface and supposedly accelerate the chances for successful implantation. The current study aims to determine the effect of Toll-like receptor 4 (TLR4) overexpression in mouse blastocysts via Let-7a downregulation using intracytoplasmic sperm injection-sperm-mediated gene transfer on embryo attachment rate. The pLenti-III-GFP-miR-Off-Let-7a vector was transmitted to oocytes derived via in vitro maturation (IVM) and in vivo oocytes by using NaOH-treated spermatozoa. Let-7a and TLR4 expression levels were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR), immunocytochemistry, and western blot analysis in both oocytes and embryos. Blastocyst adhesion on the endometrial cells was monitored by microscopic analysis. qRT-PCR results showed that Let-7a expression decreased in the IVM (GV-MII) oocytes compared to the in vivo oocyte (MII) group (p < .05). TLR4 showed a higher expression in GV-MII oocytes at both the gene and protein levels (p < .05). Following anti-miR-Let-7a transmission, the TLR4 expression level was significantly upregulated in embryos compared with the control groups (p < .05). Attachment and migration of trophoblasts cells towards endometrial cells dramatically increased compared to the control group (p < .05). Based on our results, we concluded that Let-7a might mediate embryo attachment through regulation of TLR4 expression levels.


Assuntos
Antagomirs/genética , MicroRNAs/genética , Oócitos/metabolismo , Receptor 4 Toll-Like/genética , Animais , Antagomirs/farmacologia , Blastocisto/metabolismo , Desenvolvimento Embrionário/genética , Endométrio/crescimento & desenvolvimento , Endométrio/metabolismo , Feminino , Fertilização In Vitro/métodos , Regulação da Expressão Gênica no Desenvolvimento/genética , Humanos , Camundongos , Oócitos/crescimento & desenvolvimento , Gravidez , Injeções de Esperma Intracitoplásmicas , Trofoblastos/metabolismo
11.
Epigenetics ; 15(9): 972-987, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32228351

RESUMO

DNA methylation is an epigenetic mark that plays an important role in genetic regulation in eukaryotes. Major progress has been made in dissecting the molecular pathways that regulate DNA methylation. Yet, little is known about DNA methylation variation over evolutionary time. Here we present an investigation of the variation of DNA methylation and transposable element (TE) content in species of the filamentous ascomycetes Neurospora. We generated genome-wide DNA methylation data at single-base resolution, together with genomic TE content and gene expression data, of 10 individuals representing five closely related Neurospora species. We found that the methylation levels were low (ranging from 1.3% to 2.5%) and varied among the genomes in a species-specific way. Furthermore, we found that the TEs over 400 bp long were targeted by DNA methylation, and in all genomes, high methylation correlated with low GC, confirming a conserved link between DNA methylation and Repeat Induced Point (RIP) mutations in this group of fungi. Both TE content and DNA methylation pattern showed phylogenetic signal, and the species with the highest TE load (N. crassa) also exhibited the highest methylation level per TE. Our results suggest that DNA methylation is an evolvable trait and indicate that the genomes of Neurospora are shaped by an evolutionary arms race between TEs and host defence.


Assuntos
Metilação de DNA , Epigenoma , Neurospora crassa/genética , Elementos de DNA Transponíveis , Regulação Fúngica da Expressão Gênica , Genoma Fúngico
12.
Heliyon ; 6(2): e03496, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32154399

RESUMO

Sleeve gastrectomy is a surgical technique and a leading method in metabolic surgery. Sleeve gastrectomy gained ever-increasing popularity among laparoscopic surgeons involved in bariatric surgery and has proved to be a successful method in achieving considerable weight loss in a short time. There are some disparate effects that patients may experience after sleeve gastrectomy including a reduction in BMI, weight, blood pressure, stroke, and cancer and also a significant remission in obesity-related diseases including type 2 diabetes (T2D), Non-alcoholic fatty liver (NAFLD), cardiovascular disease, obstructive sleep apnea, and craniopharyngioma-related hypothalamic obesity as well as non-obesity-related diseases such as gout, musculoskeletal problems, ovarian disorders and urinary incontinence. The most common complications of sleeve gastrectomy are bleeding, nutrient deficiencies, and leakage. There are several studies on the impact of gender and ethnic disparities on post-operative complications. This study collects state of the art of reports on sleeve gastrectomy. The aim of this study was to analyze recent studies and review the advantages and disadvantages of sleeve gastrectomy.

13.
J Mater Sci Mater Med ; 31(1): 8, 2019 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-31838602

RESUMO

Endometriosis is a common, chronic gynecological disorder associated with ongoing pelvic pain, infertility, and adhesions in reproductive age women. Current therapeutic strategies are not effective and the recurrent nature of endometriosis makes it difficult to treat. In this study, we have designed a drug delivery system to control sustained and prolonged release of curcumin in the peritoneum and pelvic cavity of a mouse model of endometriosis. Poly ε-Caprolactone (PCL) and poly ethylene glycol (PEG) polymers were used to synthesize curcumin loaded nanofibers. After scanning electron microscopy (SEM) observation of the nanofiber's morphology, we evaluated the drug release profile and in vitro degradation rate of the curcumin-loaded nanofibers. Next, we tested these nanofibers in vivo in the peritoneum of an endometriosis mouse model to determine their anti-endometriosis effects. Histological evaluations were also performed. Curcumin loaded nanofibers were successfully synthesized in the 8 and 10 wt% polymers. The release test of the curcumin-loaded nanofibers showed that approximately 23% of the loaded curcumin was released during 30 min, 35% at 24 h, and 50% at 30 days. Endometriosis was successfully induced in Balb/c mice, as noted by the observed characteristics of endometriosis in all of the mice and confirmation of endometriosis by hematoxylin and eosin (H&E) staining. In vivo experiments showed the ability of these implanted curcumin loaded nanofibers to mitigate endometriosis. We observed a considerable reduction in the endometrial glands and stroma, along with significant reduction in infiltration of inflammatory cells. Implantable curcumin loaded nanofibers successfully mitigated intraperitoneal endometriosis.


Assuntos
Anti-Inflamatórios/uso terapêutico , Curcumina/química , Curcumina/uso terapêutico , Endometriose/tratamento farmacológico , Nanofibras/química , Animais , Anti-Inflamatórios/administração & dosagem , Curcumina/administração & dosagem , Formas de Dosagem , Sistemas de Liberação de Medicamentos , Técnicas Eletroquímicas , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Estrutura Molecular
14.
J Cell Biochem ; 120(12): 19691-19698, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31297859

RESUMO

microRNAs (miRNAs) play a critical role in implantation and development of mouse embryos. In this study, we aim to evaluate the possibility of miRNAs as potential biomarkers in the blastocyst culture to assess embryo quality. We also intend to investigate whether improved clinical outcomes of vitrified embryos agree with altered miRNA expressions. Mouse embryos from in vitro fertilization were vitrified at the two-cell stage. After thawing, the embryos were individually cultured and developed to the blastocyst stage. We used quantitative real-time polymerase chain reaction to evaluate miRNA expression levels in both vitrified and fresh groups, and culture medium (CM). The fibronectin binding assay was performed to examine for blastocyst attachment. The findings showed reduced expressions of miR-16-1 (0.2 ± 0.06) and miR-Let-7a (0.65 ± 0.1) after vitrification compared to fresh embryos. We observed significant upregulation of the target genes Vav3 (4.33 ± 0.25), integrin ß-3 (Itg ß3; 4.73 ± 0.2), and Bcl2 (2.29 ± 0.16) in the vitrified embryos compared to the fresh groups. Evaluation of blastocyst CM showed upregulation of miR-Let-7a (15.68 ± 0.89), miR-16-1 (16.18 ± 0.75), and miR-15a (13.36 ± 0.73) in the vitrified group in comparison to the fresh blastocysts (P < .05). The expression levels of miR-16-1 (3.28 ± 0.63), miR-15a (5.91 ± 0.38), and miR-Let-7a (9.07 ± 0.6) in CM of the vitrified blastocysts conducted on fibronectin were significantly higher than the fresh group (P < .05).This study showed that vitrification of embryos changes implantation and proliferation biomarkers. In addition, upregulated miRNAs in CM could be potentially used for noninvasive early assessment of embryo quality.


Assuntos
Blastocisto/metabolismo , Meios de Cultura/metabolismo , Embrião de Mamíferos/metabolismo , Desenvolvimento Embrionário/genética , Regulação da Expressão Gênica no Desenvolvimento , MicroRNAs/genética , Vitrificação , Animais , Blastocisto/citologia , Técnicas de Cultura Embrionária , Implantação do Embrião , Embrião de Mamíferos/citologia , Feminino , Fertilização In Vitro , Camundongos
15.
J Cell Physiol ; 234(10): 18942-18950, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30916357

RESUMO

Proper epigenetic modifications during preimplantation embryo development are important for a successful pregnancy. We aim to investigate the putative influence of in vitro fertilization (IVF) and vitrification on DNA methylation in mouse preimplantation embryos. The study groups consisted of blastocyst-derived vitrified two-cell embryos, nonvitrified embryos, and a control group of in vivo derived blastocysts. We assessed developmental competence, global DNA methylation, relative expression levels of miR-29a/29b, and their target genes, Dnmt3a/3b. Vitrified embryos had a lower developmental rate as compared with nonvitrified embryos. There was no significant decrease in blastocyst cell numbers among studied groups, whereas there was a steady decline in DNA methylation after IVF and vitrification. The levels of miR-29a/29b upregulated in the experimental groups as compared with the control group. IVF and vitrification caused Dnmt3a/3b downregulations in blastocysts. The results of this study have suggested that a relationship exists between IVF and embryo vitrification with methylation interruptions in the blastocysts.


Assuntos
DNA (Citosina-5-)-Metiltransferases/metabolismo , Metilação de DNA/genética , Fertilização In Vitro , MicroRNAs/genética , Vitrificação , Animais , Blastocisto/metabolismo , DNA (Citosina-5-)-Metiltransferases/genética , Embrião de Mamíferos/metabolismo , Desenvolvimento Embrionário/fisiologia , Epigênese Genética/genética , Feminino , Masculino , Camundongos , MicroRNAs/biossíntese , Gravidez
16.
Andrologia ; 51(6): e13276, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30920003

RESUMO

Sperm quality can be affected by different factors including the length of incubation time between sperm preparation and intracytoplasmic sperm injection. Here, we have evaluated the level of DNA methylation and expressions of related genes in mice spermatozoa. The spermatozoa were divided into three groups: fresh, spermatozoa incubated at room temperature (RT) and 37°C for 24 hr. The sperm chromatin structure assay was used to determine the DNA fragmentation index (DFI), and DNA methylation was analysed by flow cytometry. The expression levels of DNA methylation-related genes were determined by quantitative real-time PCR (qRT-PCR). According to the results, we observed significantly higher sperm progressive motility and viability in the group incubated at RT compared to the spermatozoa incubated at 37°C (p < 0.05). Spermatozoa incubated at 37°C had a higher DFI compared to the other groups (p < 0.05), but the DNA methylation level significantly decreased (p < 0.05). qRT-PCR analysis showed increased Dnmt-1 expression in spermatozoa after 24-hr incubation at 37°C. However, there were significantly higher expression levels of Dnmt-3l, Dnmt-3a and Dnmt-3b after incubation at both RT and 37°C compared to the fresh group (p < 0.05). The 24-hr incubation period affected both sperm DNA methylation and integrity. This study indicated that incubation at RT resulted in better sperm quality.


Assuntos
Cromatina/metabolismo , Fragmentação do DNA , Metilação de DNA , Injeções de Esperma Intracitoplásmicas/métodos , Animais , Temperatura Alta/efeitos adversos , Masculino , Camundongos , Modelos Animais , Motilidade Espermática , Espermatozoides/metabolismo , Fatores de Tempo
17.
J Reprod Infant Psychol ; 37(3): 302-310, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30786750

RESUMO

Objective: The aim of this study was to assess the effect of providing massage (tactile and kinesthetic stimulation) on behavioural responses for preterm infants. Background: These infants have immature central nervous systems. Sleep-wake states during infancy have been shown to reflect severity of disease developments, and the nervous system and brain maturation. Methods: This study is a quasi-experimental study with before and after design conducted on 45 preterm infants who were admitted in neonatal intensive care unit (NICU). The subjects received massage 15 min per day for 5 days using field massage technique. Behavioural responses were measured by behavioural state, motor activity and behavioural distress. Data were obtained 10 min before and 10 min after the providing massage period. Results: An increase was observed in sleep state score after providing massage. And also, the awake, fidgeting and motor activity scores reduced after providing massage. No significant change was seen in the total behavioural distress. Conclusion: The findings suggest that providing field massage had soothing and calming effect on preterm infants and could be beneficial in nursing intervention. Nurses working in neonatal intensive care unit need to be educated on how to performing the massage on preterm infants. Abbreviation: NICU: Neonatal Intensive Care Unit.


Assuntos
Cuidado do Lactente , Recém-Nascido Prematuro , Unidades de Terapia Intensiva Neonatal , Massagem/métodos , Estudos Controlados Antes e Depois , Feminino , Idade Gestacional , Hospitais de Ensino , Humanos , Recém-Nascido , Irã (Geográfico) , Masculino , Atividade Motora , Sono
18.
J Cell Physiol ; 234(8): 13858-13866, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30605234

RESUMO

miR-224 is associated with polycystic ovary syndrome (PCOS) that is an epidemic in reproductive age women. Most studies of miR-224 have focused on in vitro analyses, whereas the in vivo effects are not widely understood. In this study, we have conducted in silico analysis and found two potential miR-224 target genes, Ptx3 and Smad4 that have roles in folliculogenesis. Because patients with PCOS have decreased numbers of follicular cells related to cell apoptosis, we also investigated two apoptotic genes, Bax and Bcl2. We used the intraovarian injection method to deliver miR-224 into a mouse model. Histological examination of the ovaries was done by fluorescent microscope. Fertilization, cleavage, and developmental competence rates were counted under a stereomicroscope and compared between the studied groups. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of miR-224 was conducted to determine the levels of the studied genes in the oocytes, cumulus cells, and blastocysts. The numbers of oocytes and fertilization rate indicated a higher apoptosis index ( p < 0.05) and increased numbers of degenerated embryos with irregular blastomeres and fragmented cytoplasm in the experimental group. RT-PCR results indicated a significant increase in miR-224 levels in the manipulated group. Of the four analyzed genes, Ptx3, Smad4, and Bcl2 had decreased levels in the transfected group, with increased Bax expression ( p < 0.05). This data showed that miR-224 negatively affected ovulation in the mouse model by decreasing Ptx3 and Smad4 expressions. The changes in Bcl2 and Bax expression levels, as apoptosis biomarkers, showed that apoptosis was a secondary outcome of the effect of miR-224.


Assuntos
Desenvolvimento Embrionário , MicroRNAs/administração & dosagem , Oócitos/metabolismo , Ovário/metabolismo , Síndrome do Ovário Policístico/genética , Animais , Biomarcadores/metabolismo , Embrião de Mamíferos/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Fluorescência Verde/metabolismo , Camundongos , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
19.
J Cell Biochem ; 120(6): 9430-9436, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30506995

RESUMO

Embryo manipulations may cause the misexpression of various genes, most of which play critical roles in the regulation of implantation. This study aimed to evaluate the effects of embryo biopsy on the expression of miR-Let-7a and its gene targets including ErbB4, Tgf-α, Itg-αv, Itg ß3 on the implantation of mouse embryo. Embryos were produced by in vitro fertilization followed by blastomere biopsy at the eight-cell stage. The effects of blastomere removal on the expression of genes ErbB4, Tgf-α, Itg αv, Itg ß3, and miR-Let-7a as well as the alteration of the blastocyst cell number were compared in both biopsied and non-biopsied groups. Finally, blastocyst attachment was assessed on culture dishes precoated with Fibronectin. The results revealed that there were no significant differences between the biopsied and non-biopsied embryos with reference to the blastocyst formation rates, the average inner cell mass, trophectoderm cell number, and percentage of attachment of blastocysts (P > 0.05). The expression of ErbB4, Itg-ß3, Itg-αv, TGF-α transcripts, and miR-Let-7a in blastocysts biopsied embryos did not differ from the non-biopsied blastocysts (P > 0.05). The results demonstrated that the preimplantation embryo development and attachment of biopsied embryos in vitro is not adversely affected by one blastomere biopsy at the eight-cell stage embryo.


Assuntos
Blastômeros/metabolismo , Implantação do Embrião/genética , Desenvolvimento Embrionário/genética , MicroRNAs/genética , Animais , Biópsia , Blastocisto/metabolismo , Embrião de Mamíferos , Feminino , Fertilização In Vitro , Regulação da Expressão Gênica no Desenvolvimento/genética , Humanos , Integrina alfa5/genética , Integrina beta3/genética , Camundongos , Gravidez , Receptor ErbB-4/genética , Fator de Crescimento Transformador alfa/genética
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