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"n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Despite advances in cancer diagnosis and treatment, survival rate of patients suffering from glioblastoma multiform (GBM) has not been significantly improved. Therefore, novel therapeutic adjuncts to routine therapies have been suggested over time. Inhibition of angiogenesis by antiangiogenic drugs is one of the new approaches to inhibit the growth of malignant cells. Microvessel density (MVD) assay is a technique performed by counting immunohistochemically-stained blood vessels. Nowadays, athymic nude mice are widely used for the establishment of xenograft tumor models in cancer research. The aim of this study was to evaluate the MVD of autochthonous xenograft models of GBM isolated from Iranian patients for use in pharmaceutical research on antiangiogenic drugs."n"nMethods: Fresh tumor samples of GBM were obtained from three patients in Cancer Institute of Tehran University of Medical Sciences in Fall of 2010 and Winter of 2011. After preliminary processing, minced tumor samples were implanted heterotopically on flanks of athymic nude mice. Two months later, the animals were sacrificed and the xenograft tumor samples were sent to the pathology laboratory. After establishing the proof of the xenograft tumor type, MVD-CD34, an endothelial cell marker, was assessed by counting hot spot areas in 22 samples."n"nResults: The mean number of microvessels in these xenograft tumor models was 30±2.1."n"nConclusion: These autochthonous xenograft models of GBM can be used in preclinical settings for research on antiangiogenic drugs regarding a pharmacogenomics-based treatment regimen for the Iranian population. Moreover, such models can be used in future studies for determining the sensitivity or resistance to antiangiogenic drugs in individualized cancer therapy.

Background & Objective: In recent years, more attention has been devoted to herbal medicines. Up to now, many compounds with therapeutic effects has been extracted from the herbs. The aim of this study is to evaluate the antioxidant and the antimicrobial effect of Rumex Alveollatus L. and to partially identify the effective compounds in this plant. Materials & Methods: Extraction was performed by using maceration method for dried flower sample. Then, the antimicrobial effect of aqueous and ethanolic extracts on eight bacterial sp. and two fungi were tested using disc diffusion method. The antioxidant effect was also determined through ferric reducing potency and phosphomolybdenum followed by total phenol determination. Finally, partial detection of bioactive compounds was conducted using chemical and calorimetric methods. Results: The results showed that ethanolic extract had the most antimicrobial effect; while aqueous extract weakly affected bacterial and fungal strains. Antioxidant experiments also revealed that ethanol extract had more antioxidant effects than aqueous extract. The most content of total phenolic compounds was found in ethanol extract. The results of the plant chemical determination showed the presence of flavonoids, alkaloids, anthraquinones, tannins, glycosides, and reducing sugars. Conclusion: Considering that few reports about the therapeutic effect of Rumex alveollatus L. has been published, this study could be considered as a valuable report about the important role of this plant on preventing infections and neutralizing oxidant agents. [ABSTRACT FROM AUTHOR]

BACKGROUND: Follicular fluid (FF) is biological fluid rich in nutrients, growth factors, hormones and may affect the sperm quality. Sperm washing has been done using conventional media in laboratory procedure so far. AIM: This study aimed to investigate the effects of FF on survival and maintenance of chromatin integrity post swim up. MATERIALS AND METHODS: Each washed semen sample was divided into two parts; the control group was incubated in the media, and the experimental groups incubated in the media containing 10% follicular fluid. Smears were prepared after 20 min, 180 min, 24 hours and no incubation times. Sperm chromatin changes like protamine, histone, DNA denaturation, sperm chromatin stability and motility were evaluated at different times. RESULTS: Incubation of sperm in the follicular fluid increased sperms with normal histone, normal chromatin protamine and sperm with normal head size (p < 0.05). CONCLUSIONS: Administration of follicular fluid into the culture media of the sperms that had been separated by swim up method could improve the sperm quality. Further studies are recommended for understanding the mechanism of the structural change of the sperm chromatin. [ABSTRACT FROM AUTHOR]

Introduction: Nowadays, freezing process is a common tool in assisted reproductive laboratories. Oxidative stress during freezing process is a major factor of sperm defect after this procedure. Despite of the fact that antioxidant compounds have an effective role on semen freezing techniques, the aim of this study was to evaluate the antioxidant effect of Calligonum on cryopreserved human sperm parameters. Materials and Methods: 10 normozoospermic patients participated in this study. Semen inclusion criteria were as follows: concentration, >60x106/ml; motility, >50%; total motile sperm count, >40x106 and strict morphology score, >10%. First of all, semen samples were frozen. Then, semen samples were washed ones with Ham's F-10 medium +10% HAS, after 24 hours of freezing. After washing, the sperm suspensions were incubated for 1 h at 37°C in medium supplemented with 10 μM/ml concentration of Calligonum extract. Morphology percent, swim up sperm count, total and forward motility of the spermatozoa were measured using computer-aided semen analyzer (CASA). Data was analyzed by one way ANOVA test. Results: The total and forward motility were significantly (p⩽0.05) higher in sperm swim up with the antioxidant (10 μM/ml) than in the control group. Data didn't show any significant difference between two groups in count and morphology percent. Conclusion: Our data reveled that CALLIGONUM extract can improve motility rate. This is an important factor in ART and affect on embryo quality. These changes are related to antioxidant properties of CALLIGONUM extract. [ABSTRACT FROM AUTHOR]

Introduction: Nowadays, spermatogonial stem cells (SSCs) cultivation has been used as an effective tool for infertility treatments, by many researchers. Oxidative conditions can be effectiveness on cell proliferation and differentiation of these cells. So, the aim of this study was establishment of oxidative stress model for antioxidant activity of some drugs investigation during SSCs in vitro culture. Materials and Methods: Neonatal NMRI male mice (3-5 day) were used for isolation of SSCs. The cell suspension was prepared by twice enzymatic digestion. The cell suspension contents were spermatogonial and Sertoli cells and treated by different doses of H2O2 logarithmic concentrations from 0-1000 μM after 24 hours. To access the optimal dose, extra doses from 10-100 μM was evaluated. After 2 hours of H2O2 treatment, viability was determined by MTT assay. The data was analyzed using SPSS software and One-way ANOVA test. Results: Our data showed that spermatogonial cells colonies appeared after 4 days of isolation. These cells expressed OCT4 and PLZF proteins. Many of spermatogonial cells were removed after using of higher doses of H2O2. The results showed that 50 μM concentration of H2O2 could induce oxidative stress in spermatogonial stem cell during in vitro culture. Conclusion: According to this study, 50 μM concentration of H2O2 can cause cell death lower than 50% of total number of cells and increase oxidative stress in cultivation of SSCs. This model is a suitable tool for studying of some new antioxidant drugs. [ABSTRACT FROM AUTHOR]

Introduction: Polycystic ovarian syndrome as a prevalent disease in women should be considered. This disease is due to an increase in the number of cysts in the ovaries and balance of oxidant and antioxidant can be affected by that. Therefore, this study is an attempt to investigate oxidative stress level in cystic ovarian tissue of mouse model. Methods and Methods: In this study, 10 NMRI female mice were divided into 2 groups: control and experimental (PCOS that was induced using single injection of estradiol valerate; 40 mg/kg). During 8 weeks after PCOS induction, body weight was measured weekly. Mice in each group were scarified and their ovarian tissue was collected. To confirm the model, histopathological study was down. Oxidative stress was measured in ovarian tissue by using DCFH-DA and fiowcytometric assay. Data was analyzed by T-test. Results: The results showed that there is a significant increasing level (p<0.05) of gaining weight in experimental group compared to control group. Ovarian tissue histopathological study was demonstrated PCO model after passing two months of induction. Fiowcytometric data reveled that oxidative stress increased in ovarian tissue of PCO group. Conclusion: The results of this study express that PCO can lead to increase oxidative stress in ovarian tissue. This increase can lead to a feedback and make more cysts in ovarian tissue of patient. [ABSTRACT FROM AUTHOR]

Introduction: Nowadays, using of vitrification to preserve of embryo is a routine technique. Re-vitrification, is applied in some situations like unprepared paitient's endometrium. In this study profiles of Bax, Bcl-2 and ErbB4 genes were evaluated in vitrified and re-vitrified mouse blastocysts. Materials and Methods: 5-8 cell embryos were collected from female mature mice, 60-62 h post hCG injection. The embryos were divided to three groups included: fresh, vitrified at blastocyst stage, vitrified (in 5-8 cells stage) and then re-vitrified at blastocyst stage. Vitrification was done using cryolock method according Kuwayama report. The relative quantification of Bax, Bcl-2 and ErbB4 genes was carried out by real time PCR. Total RNA was isolated from blastocyst embryos using QIAzol. RAN was transcribed to cDNA. Then real time PCR was performed. The data analyzed with ANOVA. Results: Our data showed that expression of Bax (pro apoptotic gene), Bcl-2 (anti apoptotic gene) had a significant difference (p<0.05) between re-vitrified and fresh embryos. Although; there was no significant difference between re-vitirified and vitrified embryos. Similar to pervious genes, ErbB4 gene expression had also a significant difference (p<0.05) between re-vitrified and fresh embryos. Re-vitrification down regulated the expression of ErbB4 (implanting gene). Although there was no significant difference between re-vitirified and vitrified embryos regarding genes expression. Conclusion: Based on our study, re-vitrification like vitrification could reduce expression of implanting gene and altered apoptotic genes expression compared to fresh embryo. [ABSTRACT FROM AUTHOR]

Introduction: Spermatogonial stem cells (SSCs) are the foundation of spermatogenesis. Sound wave especially low intensity ultrasound (LIUS) can be effective on increasing the number of cells. Materials and Methods: Isolated SSCs from neonate mice cultured in DMEM culture medium with 10% Fetal Bovine Serum. In the first phase, temperature controlled and in the second phase, SSCs stimulated by LIUS with 3 different Intensity dose (100, 200 and 300 mW/cm2) for 5 day and SSCs proliferation and colonization assessed at 71 day. Results: The LIUS treatment of mouse SSCs increased the proliferation rate and colonization of SSCs in experimental groups compared to the control group. Average of proliferation rate in 100, 200, 300 mW/cm2 and control group were 1.96±0.03, 2.26±0.03, 1.73±0.03 and 1.66±0.03, respectively. Average number of colonies in 100, 200, 300 mW/cm2 and control group were 45±1.2, 53±2.4, 28±1.2 and 20±0.8, respectively. Average diameters of colonies in 100, 200, 300 mW/cm2 and control group were 235.3±6.8 μm, 204.6±12.3 μm, 203.6±5.6 and 214.3±9.1 μm, respectively. Our results showed that there was significant increase in proliferation rate and number of colonies in experimental groups compared to control group (p<0.05), whereas there were not significant differences between groups regarding to diameter of colonies. Conclusion: These results suggested that LIUS treatment can be an efficient and cost-effective method to improve proliferation and colonization of SSCs during in vitro culture. [ABSTRACT FROM AUTHOR]

Coatings play a crucial role in the functionality of vacuum chambers in particle accelerators, serving a dual goal by efficiently facilitating pumping and mitigating electron cloud effects. However, their impact on the surface impedance of the chamber walls raises concerns, potentially affecting the machine performance and imposing limitations on achievable energies and currents. Therefore, an electromagnetic characterization is essential for a comprehensive study of accelerator structures, particularly in the context of the next-generation machines where the demand for extremely short particle bunches accentuates the importance of evaluating material responses in the very-high-frequency region. We present a technique for probing the sub-THz response of coating materials by measuring pulsed signals passing through a specifically designed waveguide, in which is placed a slab with the deposited material under test. The proposed methodology allows for a comprehensive exploration of the electromagnetic properties of the most used technical surfaces (substrate plus coatings) in accelerators under realistic conditions, providing valuable insights into their behavior in the sub-THz frequency range. The experimental data of three different Non-Evaporable Getter coating samples, prepared on a copper substrate at the CERN deposition facilities under different sputtering conditions, are discussed. The findings contribute to a deeper understanding of the complex interactions between coatings and accelerator structures, with the aim of optimizing performance and efficiency in the evolving landscape of particle acceleration technologies. The limitations and advantages of the technique are also reported.

Purpose: This study examined SSC proliferation on an epididymosome-enriched decellularized testicular matrix (DTM) hydrogel and spermatogenesis induction in azoospermic mice., Methods: Epididymosomes were extracted and characterized using SEM and western blotting. After cryopreservation, thawed SSCs were cultured in a hydrogel-based three-dimensional (3D) culture containing 10 ng/mL GDNF or 20 µg/mL epididymosomes. SSCs were assessed using the MTT assay, flow cytometry, and qRT-PCR after two weeks of culture. The isolated SSCs were microinjected into the efferent ducts of busulfan-treated mice. DiI-labeled SSCs were followed, and cell homing was assessed after two weeks. After 8 weeks, the testes were evaluated using morphometric studies and immunohistochemistry., Results: The expression of PLZF, TGF-β, and miR-10b did not increase statistically significantly in the 3D + GDNF and 3D + epididymosome groups compared to the 3D group. Among the groups, the GDNF-treated group exhibited the highest expression of miR-21 (*P < 0.05). Caspase-3 expression was lower in the epididymosome-treated group than in the other groups (***P < 0.001). Compared to the 3D and negative control groups, the 3D + epididymosomes and 3D + GDNF groups showed an increase in spermatogenic cells. Immunohistochemical results confirmed the growth and differentiation of spermatogonial cells into spermatids in the treatment groups., Conclusion: The DTM hydrogel containing 20 µg/mL epididymosomes or 10 ng/mL GDNF is a novel and safe culture system that can support SSC proliferation in vitro to obtain adequate SSCs for transplantation success. It could be a novel therapeutic agent that could recover deregulated SSCs in azoospermic patients., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

We report a study on the hydrogen bonding mechanisms of three aliphatic alcohols (2-propanol, methanol, and ethanol) and one diol (ethylene glycol) in water solution using a time-domain ellipsometer in the THz region. The dielectric response of the pure liquids is nicely modeled by the generalized Debye-Lorentz equation. For binary mixtures, we analyze the data using a modified effective Debye model, which considers H-bond rupture and reformation dynamics and the motion of the alkyl chains and of the OH groups. We focus on the properties of the water-rich region, finding anomalous behavior in the absorption properties at very low solute molar concentrations. These results, first observed in the THz region, are in line with previous findings from different experiments and can be explained by taking into account the amphiphilic nature of the alcohol molecules.

The use of a combination of three-drug regimen has improved HIV-1 infected patients' life span and quality; however the emergence of drug-resistant strains remains a main problem. Reverse transcriptase inhibitors (RTIs) consist of a main part of highly active anti-retroviral therapy (HAART) regimen. The present study aimed to investigate resistant mutations to RTI drugs in both treatment naïve and under treatment HIV patients in Mashhad city, north-eastern Iran. RNA was extracted from sera of 22 treatment naïve and 22 under treatment patients. The mean age of under treated and treatment naive groups were 38.5±6.7 and 40.8±7.9 respectively. cDNA was synthesized and amplified with Nested PCR assay targeting specific sequences of RT gene. The PCR products were sent for sequencing. Bidirectional sequencing results were analysed using HIV drug resistance database supplied by Stanford University (HIV Drug Resistance Database, https://hivdb.stanford.edu). Among under treatment patients 10 out of 22 (45%) had at least one high-level resistance mutation which was higher than high level resistance mutation rate among treatment naive cases (P<0.01). Detected resistance mutations were as follows: K101E, K103N, K103E, V106M, V108I, E138A, V179T, Y181C, M184V, Y188L, Y188H, Y188F, G190A, L210W, T215F, T215Y, K219Q, and P225H. A high level of resistance mutations to RT inhibitors was observed that causes drug resistance especially against lamivudine (3TC). Such mutations should be considered as probable responsible for therapeutic failure. Serial surveillance studies of circulating drug resistance mutations are recommended., Competing Interests: The author declared no conflict of interest.

Telocytes are interstitial cells found in different tissues, including cardiac stem cell niches. The purpose of this study was to investigate the response of the telocytes to the cardiac growth that occurs in response to resistance and endurance exercise trainings using rats distributed into control, endurance, and resistance training groups. Results revealed that the ratio of heart weight to body weight, cardiomycyte number, cardiomyocyte area, thickness of the left ventricular wall were significantly higher in the training groups compared to the control group. We observed increment in the cardiomyocytes surface area and thickness of the left ventricular wall in the resistance-training group than endurance-training group. We conclude that both resistance and endurance exercise trainings will lead to an increased number of cardiac telocytes, consequently, promote activity of the cardiac stem cells, and results in physiological cardiac growth, and this response does not seem to depend on the type of exercise., (© 2023. The Author(s).)

Using a customized time-domain ellipsometer operating in the THz range, the molecular dynamics of a liquid binary solution based on water and isopropyl alcohol (2-propanol) is investigated. The setup is capable of detecting small changes in the optical properties of the mixture within a single measurement. The complex dielectric response of samples with different concentrations is studied through the direct measurement of the ellipsometric parameters. The results are described using an effective Debye model, from which the relaxation parameters associated with different activation energies can be consistently extracted. Significant deviations between experimental data and the theoretical expectations at an intermediate volume percentage of 2-propanol in water are observed and interpreted as produced by competing effects: the creation/destruction of hydrogen bonding on the one hand, and the presence of cluster/aggregation between water and alcohol molecules on the other.

Background: Open cranial vault reconstruction is the standard technique of craniosynostosis correction that may cause significant blood loss., Objectives: The current study aimed at comparing the effect of tranexamic acid (TXA), controlled hypotension, and their combination on perioperative blood loss and transfusion requirement in craniosynostosis surgery., Methods: The present randomized, double-blind clinical trial was conducted on 75 infants referred for craniosynostosis surgery during 2017 - 2018. Ten minutes before the start of surgery, 10 mg/kg of TXA was administered intravenously to patients in the first group (TXA group). In the second group, patients were subjected to the controlled hypotension anesthesia (CHA) using intravenous remifentanil 0.1 μ/kg (CHA group). In the third group, the patients underwent CHA similar to that of the second group, along with intravenous injection of 10 mg/kg of TXA (CHA-TXA group). Then, patients' mean arterial pressure (MAP), heart rate (HR), total blood loss, and transfusion volume were evaluated and recorded., Results: The results of the present study revealed that although the changes in MAP and HR parameters over time (three hours after surgery) were significant in all three groups, the lowest decrease was observed in the CHA-TXA group (P-value < 0.05). In addition, the total perioperative blood loss in the CHA-TXA group with the mean of 181.20 ± 82.71 cc was significantly less than the total perioperative blood loss in the CHA and TXA groups with the means of 262.00 ± 104.04 cc and 212.80 ± 80.75 cc, respectively (P-value < 0.05). Moreover, the transfusion volume in the CHA-TXA group with the mean of 112.40 ± 53.50 cc was significantly lower than the transfusion volume in the CHA and TXA groups with the means of 174.00 ± 73.93 cc and 160.63 ± 59.35 cc, respectively (P-value < 0.05). In contrast, the total blood loss and transfusion volume were not significantly different between the CHA and TXA groups (P-value > 0.05)., Conclusions: According to the results of the present study, although the administration of TXA alone could effectively prevent blood loss and was associated with fewer transfusion requirements, the combination of this approach with hypotensive anesthesia resulted in more reduction in perioperative blood loss and transfusion volume as well as better hemodynamic stability., Competing Interests: Conflict of Interests: The authors declare they have no conflict of interests., (Copyright © 2023, Author(s).)

Aims: This study aimed to explore the effect of epididymosomes on the proliferative efficiency of spermatogonial stem cells (SSCs) in vitro and the resumption of spermatogenesis in the azoospermic mice., Main Methods: The epididymosomes were extracted from the epididymis and characterized. SSCs were cultured in 2D (two-dimensional) and hydrogel-based 3D culture in the presence of 20 μg/mL epididymosome or 10 ng/mL GDNF. After two weeks of culture, the proliferation and purity of the separated SSCs were evaluated using the MTT test and flow cytometry, respectively. qRT-PCR was used to analyze PLZF, caspase-3, TGF-β, miR-10b, and miR-21 expression levels. Then, SSCs grown in the 3D culture system were labeled by DiI and transplanted into azoospermic mice via the efferent duct. After 2 weeks, tracing of DiI and cell homing were evaluated. Subsequently, histomorphometric studies and immunohistochemistry analysis were performed in testes after eight weeks of transplantation., Key Findings: The expression of PLZF, TGF-β, miR-10b, and miR-21 increased significantly (*p < 0.05) in the 3D + GDNF and 3D + epididymosomes groups than in the 2D group. Transplanted SSCs migrated into the seminiferous tubules of recipient mice and the number of spermatogenic cells and protein expression of PLZF, SCP3 and ACRBP in the 3D + GDNF and 3D + epididymosomes groups were considerably higher (∗ ∗ ∗ p < 0.001) compared to the azoospermic group., Significance: This finding indicates that culturing SSCs on decellularized testicular matrix (DTM) hydrogel with 10 ng/mL GDNF or 20 μg/mL epididymosomes could lead to an increase in SSCs proliferation which provides a sufficient number of SSCs for successful transplantation in azoospermic mice., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest., (Copyright © 2022. Published by Elsevier Inc.)

Although numerous studies have investigated the molecular basis of male infertility, various aspects of this area have remained uncovered. Over the past years, researchers have reported the significant potential of miRNAs in posttranscriptional regulatory roles. By targeting mRNAs, these notable molecules can modulate the processes related to male infertility. On the other side, the outstanding potential of male germline stem cells, SSCs, includes their application in infertility treatment. SSCs retain normal spermatogenesis and fertility by adjusting both SSC self-renewal and differentiation. Therefore, for the characterization and manipulation of SSCs, effective and efficient in vitro culture methods are essential in supporting their maintenance and development. In this regard, the present investigation was undertaken to evaluate the impact of one of the recently conspicuous miRNAs, miR-106b, in SSCs enrichment. As a result, we first found that the SSCs induced with miR-106b-5p highly express TGF-β1, which is known as a regulator of epigenetic modifiers and downstream genes. We next sought to show that self-renewal markers, including c-Myc, Oct-4, and Sox2, are increased in the induced SSC group. The intended miRNA also induced the inhibitor of differentiation 4 (ID4) and aided to remain unmethylated in SSCs. Additionally, for the tumorigenicity possibility of the manipulation, we indicated that PTEN, a tumor-suppressor gene, expressed remarkably in the induced SSCs. In conclusion, our findings showed that miR-106b-5p enhances the proliferative potential of SSCs, making it a substantial factor for therapeutic strategies of male infertility., (© 2022. Society for Reproductive Investigation.)

N-methyl-d-aspartate (NMDA) modulates the spermatogenesis process through stimulating the steroid hormone biosynthesis. The aim of this study was to evaluate the effects of NMDA receptors agonists (d-Serine) and antagonists (MK801) on spermatogonia differentiation on decellularization testicular matrix (DTM) hydrogel scaffold. Four treatment groups were planned: 2D + D-Serine, 3D + D-Serine, 2D + MK801, and 3D + MK801. Results showed that cell viability was significantly decreased after 48 h in the 3D + D-Serine group and after 24 and 48 h in the 3D + MK801 group compared to the controls. The spermatogonia proliferation after two, four, and eight weeks was significantly increased in the 3D + D-Serine culture, while it was significantly reduced in the 2D + MK801 and 3D + MK801 groups after four and eight weeks. Real-time PCR results demonstrated that pre-meiotic gene (Plzf) expression was significantly increased only in the 3D + D-Serine culture compared to the control groups after four weeks of culture. The meiotic gene (Sycp3) expression was significantly increased in the 2D + D-Serine and 3D + D-Serine compared to the 2D controls after four and eight weeks. The post-meiotic gene (Tnp1) level in the 3D + D-Serine was significantly higher than the other groups. Flow-cytometry results indicated that the protein expression of Plzf (after four and eight weeks), Sycp3 (after eight weeks), and Tnp1 (after eight weeks) in the d-Serine-treated groups was significantly increased compared with the 2D control groups. There were not any significant changes in the gene expression of spermatogenic-related markers in MK801 culture media. However, a significant decrease in the protein levels of Plzf after eight weeks and Sycp3 after four and eight weeks was observed. In conclusion, the addition of NMDARs agonists (d-Serine) could be used to regulate the differentiation of spermatogonia in the 3D culture system., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Objective: According to the mounting data, microRNAs (miRNAs) may play a key role in reprogramming. miR-106b, is considered as an enhancer in reprogramming efficiency. Based on induced pluripotent stem cells (iPSCs), cell treatments have a huge amount of potential. One of the main concerns about using iPSCs in therapeutic settings is the possibility of tumor formation. It is hypothesized that a procedure that can reprogram cells with less genetic manipulation reduces the possibility of tumorigenicity., Materials and Methods: In this experimental study, miR-106b-5p transduced by pLV-miRNA vector into mice isolated spermatogonial stem cells (SSCs) to achieve iPS-like cells. Then the transduced cells were cultured in specific conditions to study the formation of three germ layers. The tumorigenicity of these iPS-like cells was investigated by transplantation into male BALB/C mice., Results: We show that SSCs can be successfully reprogrammed into induced iPS-like cells by pLV-miRNA vector to transduce the hsa-mir-106b-5p into SSCs and generating osteogenic, neural and hepatoblast lineage cells in vitro as a result of pluripotency. Although these iPS-like cells are pluripotent, they cannot form palpable tumors in vivo., Conclusion: These results demonstrate that infection of hsa-mir-106b-5p into SSCs can reprogram them into iPSCs, and advanced germ cell lineages without tumorigenicity. Also, a novel approach for studying the generation of iPSCs, and the application of iPS or iPS-like cells in regenerative medicine is presented.

Mesenchymal stem cells can be differentiated into tissue-specific cells. MicroRNAs (miRNAs) regulate the translation of mRNAs involved in the growth and development of a variety of cells, including primordial germ cells (PGCs). This study evaluated male germ cell differentiation from human MSCs by miR-106b. The MSCs were obtained from human adipose tissue. The differentiation of MSCs into PGCs was accomplished by transfection of a lentiviral vector expressing miR-106b. MSCs were treated with bone morphogenic factor 4 as a control and also as a putative inducer of PGC differentiation. PGC was differentiated into spermatogonial-like cells by retinoic acid. Moreover, Dazl, Plzf, Stra8, Gfra, and Thy1 gene expressions were investigated using real-time PCR. Our results showed that Dazl, Plzf, and Stra8 genes that were treated with BMP4 and miR-106b did not show any significant difference, meaning that miR-106b, like BMP4, is able to differentiate PGC cells from MSCs. In spermatogonial-like cells, Thy1 was significantly unregulated in both the miR-106b and BMP4 groups. Our findings showed that miR-106b regulates the differentiation of MSCs into PGCs. miR-106b influences on the expression of Dazl, Plzf, and Stra8 genes in PGC and Gfra, Stra8, and Thy1 genes., (© 2022. The Society for In Vitro Biology.)

Ellipsometry is extensively used in the optical regime to investigate the properties of many materials as well as to evaluate with high precision the surface roughness and thickness of thin films and multilayered systems. Due to the inherent non-coherent detection technique, data analyses in optical ellipsometry tend to be complicated and require the use of a predetermined model, therefore indirectly linking the sample properties to the measured ellipsometric parameters. The aim of this tutorial is to provide an overview of terahertz (THz) time-domain ellipsometry, which is based instead on a coherent detection approach and allows in a simple and direct way the measurement of the material response. After giving a brief description of the technology presently used to generate and detect THz radiation, we introduce the general features of an ellipsometric setup operating in the time domain, putting in evidence similarities and differences with respect to the classical optical counterpart. To back up and validate the study, results of THz ellipsometric measurements carried out on selected samples are presented.

Background: Recent years have brought notable progress in raising the efficiency of the reprogramming technique so that approaches have evolved from known transgenic factors to only a few miRNAs. Nevertheless, there is a poor understanding of both the key factors and biological networks underlying this reprogramming. The present study aimed to investigate the potential of miR-106b-5p in regulating spermatogonial stem cells (SSCs) to induced pluripotent stem cell (iPSC)-like cells., Methods: We used SSCs because pluripotency is inducible in SSCs under defined culture conditions, and they have a few issues compared to other adult stem cells. As both signaling and post-transcriptional gene controls are critical for pluripotency regulation, we traced the expression of Oct-4, Sox-2, Klf-4, c-Myc, and Nanog (OSKMN). Besides, we considered miR-106b-5p targets using bioinformatic methods., Results: Our results showed that transfected SSCs with miR-106b-5p increased the expression of the OSKMN factors, which was significantly more than negative control groups. Moreover, using the functional miRNA enrichment analysis, online tools, and databases, we predicted that miR-106b-5p targeted a signaling pathway gene named MAPK1/ERK2, related to regulating stem cell pluripotency., Conclusion: Together, our data suggest that miR-106b-5p regulates the reprogramming of SSCs into iPSC-like cells. Furthermore, noteworthy progress in the in vitro development of SSCs indicates promise reservoirs and opportunities for future clinical trials.

We report the design, fabrication and experimental validation of a THz all-dielectric multi-mode q-plate having a fixed rate of change of the optical axis. The device consists of space-variant birefringent slabs manufactured by 3D printing using melt-extruded Acrylonitrile Butadiene Styrene (ABS). The desired form birefringence is analytically evaluated and experimentally measured by the THz time domain spectroscopy technique. The manufactured q-plate design is characterized using a polarization-sensitive imaging setup. The full electric field spatial maps are acquired from the beam propagating through the q-plate. The device enables the realization of both radial and azimuthal vector beams at discrete frequency intervals by controlling the space-dependent orientation of the ordinary and extraordinary axes in the transverse plane with a multi-mode sequence.

We report on the realisation of a customized THz time domain spectroscopic ellipsometer (THz-TDSE) based on fiber-coupled photoconductive antennas, operating in a wide range of incident angles and allowing also standard transmission spectroscopy without any optical realignment. To ensure accurate parameter extraction for a broad range of materials, we developed a fast and effective algorithm-assisted method to calibrate the setup and compensate for the nonideality in the response of the THz system. The procedure allows to minimise errors induced by imperfect response of the antennas and polarizers, imprecise setting of the impinging and receiving angles in the goniometric mechanical arms, and unavoidable mismatches in the THz beam optics. Differently from other calibration methods applied in the literature, our approach compares in time domain the ellipsometric derived electric field s- and p-polarised components at a given angle of incidence with the reconstructed ones, attained by using the complex dielectric function of a known sample. The calibrated response is determined with high precision by setting the system in transmission mode. In order to validate the technique, ellipsometric measurements have been carried out at various angle of incidences on a number of materials both in solid and liquid form, and their data compared with what obtained by conventional THz spectroscopy. Results show that THz-TDSE accompanied with an accurate calibration procedure is an effective technique for material characterization, especially in case of samples with a high absorption rate that are not easily investigated through transmission measurements., (© 2022. The Author(s).)

Background: The main purpose of this study was to investigate the effect of D-serine (DS) and Dizocilpine (MK-801) on the proliferation of spermatogonial stem cells (SSCs) in two-dimensional (2D) and three-dimensional (3D) culture systems., Methods and Results: The SSCs of male NMRI mice were isolated by enzymatic digestion and cultured for two weeks. Then, the identity of SSCs was validated by anti-Plzf and anti-GFR-α1 antibodies via immunocytochemistry (ICC). The proliferation capacity of SSCs was evaluated by their culture on a layer of the decellularized testicular matrix (DTM) prepared from mouse testis, as well as two-dimensional (2D) with different mediums. After two weeks of the initiation of proliferation culture on 3D and 2D medium, the pre-meiotic at the mRNA and protein levels were evaluated via qRT-PCR and flow cytometry methods, respectively. The results showed that the proliferation rate of SSCs in 3D culture with 50 mM glutamic acid and 20 mM D-serine was significantly different from other groups after 14 days treatment. mRNA expression levels of promyelocytic leukemia zinc finger (Plzf) in 3D cultures supplemented by 20 mM D-serine and 50 mM glutamic acid were considerably higher than the 3D control group (p < 0.001). The flow cytometry analysis revealed that the amount of Plzf in the 2D-culture groups of SSCs with 20 mM MK-801 was considerably lower compared to the 2D-culture control group (p < 0.001)., Conclusions: This study indicated that decellularized testicular matrix supplemented with D-serine and glutamic acid could be considered a promising vehicle to support cells and provide an appropriate niche for the proliferation of SSCs., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Our previous study indicated that Potentilla reptans root has a preconditioning effect by its antioxidant and anti-apoptotic effects in an isolated rat heart ischemia/reperfusion (IR) model. In the present study, we investigated the post-conditioning cardio-protective effects of Potentilla reptans and its active substances., Methods: The ethyl acetate fraction of P. reptans root (Et) was subjected to an IR model under 30 min of ischemia and 100 min of reperfusion. To investigate the postconditioning effect, Et was perfused for 15 min at the early phase of reperfusion. RISK/SAFE pathway inhibitors, 5HD and L-NAME, were applied individually 10 min before the ischemia, either alone or in combination with Et during the early reperfusion phase. The hemodynamic factors and ventricular arrhythmia were calculated during the reperfusion. Oxidative stress, apoptosis markers, GSK-3β and SGK1 proteins were assessed at the end of experiments., Results: Et postconditioning (Etpost) significantly reduced the infarct size, arrhythmia score, ventricular fibrillation incidence, and enhanced the hemodynamic parameters by decreasing the MDA level and increasing expression of Nrf2, SOD and CAT activities. Meanwhile, Etpost increased the BCl-2/BAX ratio and decreased Caspase-3 expression. The cardioprotective effect of Etpost was abrogated by L-NAME, Wortmannin (a PI3K/Akt inhibitor), and AG490 (a JAK/STAT3 inhibitor). Finally, Etpost reduced the expression of GSK-3β and SGK1 proteins pertaining to the IR group., Conclusion: P. reptans reveals the post-conditioning effects via the Nrf2 pathway, NO release, and the RISK/SAFE pathway. Also, Etpost decreased apoptotic indexes by inhibiting GSK-3β and SGK1 expressions. Hence, our data suggest that Etpost can be a suitable natural candidate to protect cardiomyocytes during reperfusion injury., (© 2021. The Author(s).)

Objective: Spermatogonial stem cells (SSCs) are able to form embryonic stem-like cells (ES-like cells) and embryonic bodies (EBs). Low-intensity ultrasound stimulation (LIUS) has positive effects on the growth and differentiation of the different cells. In this study, we tried to investigate the effects of LIUS on SSC differentiation to ES-like cells., Materials and Methods: SSCs were isolated from neonatal mice and their identification was confirmed by tracking of PLZF, Oct-4, and C-Kit proteins. The SSCs and Sertoli cells were co-cultured in DMEM/F12 supplemented with 15% FBS and LIF. SSCs stimulated by LIUS with 200mW/CM 2 intensity. Characterization of obtained ES-like cells was confirmed with Sox2, Oct-4, and SSEA-1 immunofluorescence staining. Also, real-time PCR was performed to analyse the expression of c-Myc and Nanog genes in ES-Like Cells and Stra8, Piwil2 and Plzf genes in SSCs after 21 days of the in vitro culture., Results: Our results showed c-Kit, PLZF and Oct-4 proteins were expressed positively in SSCs and Sox2, Oct-4, SSEA-1 in the ES-like cells by immunocytochemistry. The results of flow cytometry showed a significant increase in expression of c-Myc and Nanog in ES-like cells compared to SSCs (p<.05), whereas the Stra8, Piwil2, and Plzf became down-regulated during 21 days of culture. ES-like markers cell SSEA-1, Sox2 and Oct-4 were increased in the LIUS group compared to the control group (p<.05)., Conclusion: The results indicated that ES-like cells with pluripotency characteristics were derived from SSCs., (Copyright © 2020 Asociación Española de Andrología, Medicina Sexual y Reproductiva. Publicado por Elsevier España, S.L.U. All rights reserved.)