Search results for: porcine oocyte.

Authors: M. Areekijseree, W. Pongsawat, M. Pumipaiboon, C. Thepsithar, S. Sengsai, T. Chuen-Im

Abstract:

Morphological interaction of porcine cumulus-oocyte complexes (pCOCs) was investigated on in vitro condition using electron microscope (SEM and TEM). The totals of 1,923 oocytes were round in shape, surrounded by Zona pellucida with layer of cumulus cells ranging between 59.29-202.14 μm in size. They were classified into intact-, multi-, partial cumulus cell layer oocyte, and completely denuded oocyte, at the percentage composition of 22.80% 32.70%, 18.60%, and 25.90 % respectively. The pCOCs classified as intact- and multi cumulus cell layer oocytes were further culturing at 37°C with 5% CO2, 95% air atmosphere and high humidity for 44 h in M199 with Earle’s salts supplemented with 10% HTFCS, 2.2 mg/mL NaHCO3, 1 M Hepes, 0.25 mM pyruvate, 15 μg/mL porcine follicle-stimulating hormone, 1 μg/mL LH, 1μg/mL estradiol with ethanol, and 50 μg/mL gentamycin sulfate. On electron microscope study, cumulus cells were found to stick their processes to secrete substance from the sac-shape end into Zona pellucida of the oocyte and also communicated with the neighboring cells through their microvilli on the beginning of incubation period. It is believed that the cumulus cells communicate with the oocyte by inserting the microvilli through this gap and embedded in the oocyte cytoplasm before secreting substance, through the sac-shape end of the microvilli, to inhibit primary oocyte development at the prophase I. Morphological changes of the complexes were observed after culturing for 24-44 h. One hundred percentages of the cumulus layers were expanded and cumulus cells were peeling off from the oocyte surface. In addition, the round-shape cumulus cells transformed themselves into either an elongate shape or a columnar shape, and no communication between cumulus neighboring cells. After 44 h of incubation time, diameter of oocytes surrounded by cumulus cells was larger than 0 h incubation. The effect of hormones in culture medium is exerted by their receptors present in porcine oocyte. It is likely that all morphological changes of the complexes after hormone treatment were to allow maturation of the oocyte. This study demonstrated that the association of hormones in M199 could promote porcine follicle activation in 44 h in vitro condition. This culture system should be useful for studying the regulation of early follicular growth and development, especially because these follicles represent a large source of oocytes that could be used in vitro for cell technology.

Keywords: Cumulus cells, electron microscopy (SEM and TEM), in vitro, porcine oocyte.

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Authors: D. Biswas, Sang H. Hyun

Abstract:

In mammalian reproductive tract, the oviduct secretes huge number of growth factors and cytokines that create an optimal micro-environment for the initial stages of preimplantation embryos. Secretion of these growth factors is stage-specific. Among them, VEGF is a potent mitogen for vascular endothelium and stimulates vascular permeability. Apart from angiogenesis, VEGF in the oviduct may be involved in regulating the oocyte maturation and subsequent developmental process during embryo production in vitro. In experiment 1, to evaluate the effect of VEGF during IVM of porcine COC and subsequent developmental ability after PA and SCNT. The results from these experiments indicated that maturation rates among the different VEGF concentrations were not significant different. In experiment 2, total intracellular GSH concentrations of oocytes matured with VEGF (5-50 ng/ml) were increased significantly compared to a control and VEGF group (500 ng/ml). In experiment 3, the blastocyst formation rates and total cell number per blastocyst after parthenogenesis of oocytes matured with VEGF (5-50 ng/ml) were increased significantly compared to a control and VEGF group (500 ng/ml). Similarly, in experiment 4, the blastocyst formation rate and total cell number per blastocyst after SCNT and IVF of oocytes matured with VEGF (5 ng/ml) were significantly higher than that of oocytes matured without VEGF group. In experiment 5, at 10 hour after the onset of IVF, pronuclear formation rate was evaluated. Monospermy was significantly higher in VEGF-matured oocytes than in the control, and polyspermy and sperm penetration per oocyte were significantly higher in the control group than in the VEGFmatured oocytes. Supplementation with VEGF during IVM significantly improved male pronuclear formation as compared with the control. In experiment 6, type III cortical granule distribution in oocytes was more common in VEGF-matured oocytes than in the control. In conclusion, the present study suggested that supplementation of VEGF during IVM may enhance the developmental potential of porcine in vitro embryos through increase of the intracellular GSH level, higher MPN formation and increased fertilization rate as a consequence of an improved cytoplasmic maturation.

Keywords: angiogenesis, GSH, monospermy, VEGF

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Authors: Mohd Sukri Hassan, Ahlam Inayatullah Badrul Munir, M. Husaini A. Rahman

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Halal authentication and verification in supplement capsules are highly required as the gelatine available in the market can be from halal or non-halal sources. It is an obligation for Muslim to consume and use the halal consumer goods. At present, real-time polymerase chain reaction (RT-PCR) is the most common technique being used for the detection of porcine and bovine DNA in gelatine due to high sensitivity of the technique and higher stability of DNA compared to protein. In this study, twenty samples of supplements capsules from different products with different Halal logos were analyzed for porcine and bovine DNA using RT-PCR. Standard bovine and porcine gelatine from eurofins at a range of concentration from 10^-1 to 10^-5 ng/µl were used to determine the linearity range, limit of detection and specificity on RT-PCR (SYBR Green method). RT-PCR detected porcine (two samples), bovine (four samples) and mixture of porcine and bovine (six samples). The samples were also tested using FT-IR technique where normalized peak of IR spectra were pre-processed using Savitsky Golay method before Principal Components Analysis (PCA) was performed on the database. Scores plot of PCA shows three clusters of samples; bovine, porcine and mixture (bovine and porcine). The RT-PCR and FT-IR with chemometrics technique were found to give same results for porcine gelatine samples which can be used for Halal authentication.

Keywords: Halal, real-time PCR, gelatin, FTIR and chemometrics.

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Authors: Marcela Capcarova, Adriana Kolesarova, Marina Medvedova, Peter Petruska, Alexander V. Sirotkin

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The aim of this study was to determine the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), total antioxidant status (TAS) and accumulation of Hsp70 in porcine ovarian granulosa cells after deoxynivalenol (DON) and zearalenone (ZEA) exposure in vitro. Porcine ovarian granulosa cells were incubated with DON/ZEA administrations as follows: group A (10/10 ng/mL), group B (100/100 ng/mL), group C (1000/1000 ng/mL), and the control group without any additions for 24h. In this study mycotoxins developed stress reaction of porcine ovarian granulosa cells and increased accumulation of Hsp70 what resulted in increasing activities of SOD and GPx in groups with lower doses of mycotoxins. High dose of DON and ZEA had opposite effect on GPx activity than the lower doses. Slight increase in TAS of porcine granulosa cells was observed after mycotoxins exposure. These results contribute towards the understanding of cellular stress and its response.

Keywords: Deoxynivalenol, zearalenone, antioxidants, Hsp70, granulosa cells.

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Authors: J. Sepitka, J. Lukes, J. Reznicek

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Hysitron TriboIndenterTM TI 950 system has been used for studying the local viscoelastic properties of porcine intervertebral disc end plate by means of nanoscale mechanical dynamic analysis. The specimen of an endplate was cut from fresh porcine vertebra dissected from 16 month animal. The lumbar spine motion segments were dissected and 5 millimeter thick plates of vertebral body, endplate and annulus fibrosus were prepared for nanoindentation. The surface of the sample was kept in physiological solution during nanoindentation experiment. We obtained mechanical characteristics of different areas of native endplate (endplate middle and vertebra and annulus fibrosus boundary).

Keywords: nanoindentation, DMA, endplate, cartilage

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Authors: K.Ting, K. T. Chen, Y. L. Su, C. J. Chang

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In medical therapy, laser has been widely used to conduct cosmetic, tumor and other treatments. During the process of laser irradiation, there may be thermal damage caused by excessive laser exposure. Thus, the establishment of a complete thermal analysis model is clinically helpful to physicians in reference data. In this study, porcine liver in place of tissue was subjected to laser irradiation to set up the experimental data considering the explored impact on surface thermal field and thermal damage region under different conditions of power, laser irradiation time, and distance between laser and porcine liver. In the experimental process, the surface temperature distribution of the porcine lever was measured by the infrared thermal imager. In the part of simulation, the bio heat transfer Pennes-s equation was solved by software SYSWELD applying in welding process. The double ellipsoid function as a laser source term is firstly considered in the prediction for surface thermal field and internal tissue damage. The simulation results are compared with the experimental data to validate the mathematical model established here in.

Keywords: laser infrared thermal imager, bio-heat transfer, double ellipsoid function.

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Authors: Caighley Logan, Suzzanne McColl

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The outcome of fire related fatalities, along with other research, has found fires can have a detrimental effect to the mineral and crystalline structures within bone. This study focused on the mineral and crystalline structures within porcine bone samples to analyse the changes caused, with the intent of effectively ‘reverse engineering’ the data collected from burned bone samples to discover what may have happened. Using Fourier Transform Infrared (FTIR), and X-Ray Fluorescence (XRF), the data were collected from a controlled source of intense heat (muffle furnace) and an open fire, based in a living room setting in a standard size shipping container (2.5 m x 2.4 m) of a similar temperature with a known ignition source, a gasoline lighter. This approach is to analyse the changes to the samples and how the changes differ depending on the heat source. Results have found significant differences in the levels of remaining minerals for each type of heat/burning (p =< 0.001), particularly Phosphorus and Calcium, this also includes notable additions of absorbed elements and minerals from the surrounding materials, i.e., Cerium (Ce), Bromine (Br) and Neodymium (Ne). The analysis techniques included provide validated results in conjunction with previous studies.

Keywords: Forensic anthropology, thermal alterations, porcine bone, FTIR, XRF.

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Authors: Young Geun Kim, Tae Woo Lee, Changmin Yeo, Jung min Yoo, Yeo Jin Kang, Tack-Joong Kim, Byungjo Jung, Ji Hun Cha, Chan Hoi Hur, Dong-Sup Kim, Ki Jung Park, Han Sung Kim

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The intention of this lessons is to assess the probability of optical coherence tomography (OCT) for biometric recognition. The OCT is the foundation on an optical signal acquisition and processing method and has the micrometer-resolution. In this study, we used the porcine skin for verifying the abovementioned means. The porcine tissue was sound acknowledged for structural and immunohistochemical similarity with human skin, so it could be suitable for pre-clinical trial as investigational specimen. For this reason, it was tattooed by the tattoo machine with the tattoo-pigment. We detected the pattern of the tattooed skin by the OCT according to needle speed. The result was consistent with the histology images. This result showed that the OCT was effective to examine the tattooed skin section noninvasively. It might be available to identify morphological changes inside the skin.

Keywords: mechanical skin damage, optical coherence tomography, tattooed skin

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Authors: Seunghwan Lee

Abstract:

The lubricating properties of commercially available mucins originating from different animal organs, namely bovine submaxillary mucin (BSM) and porcine gastric mucin (PGM), have been characterized at polymeric surfaces for biomedical applications. Atomic force microscopy (AFM) and pin-on-disk tribometry have been employed for tribological studies at nanoscale and macroscale contacts, respectively. Polystyrene (PS) was employed to represent ‘rigid’ contacts, whereas poly(dimethylsiloxane) (PDMS) was employed to represent ‘soft contacts’. To understand the lubricating properties of mucins in correlation with the coverage on surfaces, adsorption properties of mucins onto the polymeric substrates have been characterized by means of optical waveguide light-mode spectroscopy (OWLS). Both mucins showed facile adsorption onto both polymeric substrates, but the lubricity was highly dependent upon the pH change between 2 and 7.

Keywords: Bovine submaxillary mucin (BSM), Porcine Gastric Mucin (PGM), lubricity, biomedical.

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Authors: M. Petchsangsai, N. Wonglertnirant, T. Rojanarata, P. Opanasopit, T. Ngawhirunpat

Abstract:

The objective of the present study was to evaluate the potential of hollow microneedles for enhancing the transdermal delivery of Bovine Serum Albumin (MW~66,000 Da)-Fluorescein Isothiocyanate (BSA-FITC) conjugate, a hydrophilic large molecular compound. Moreover, the effect of different formulations was evaluated. The series of binary mixtures composed of propylene glycol (PG) and pH 7.4 phosphate buffer solution (PBS) was prepared and used as a medium for BSA-FITC. The results showed that there was no permeation of BSA-FITC solution across the neonatal porcine skin without using hollow microneedles, whereas the cumulative amount of BSA-FITC released at 8 h through the neonatal porcine skin was about 60-70% when using hollow microneedles. Furthermore, the results demonstrated that the higher volume of PG in binary mixtures injected, the lower cumulative amount of BSA-FITC released and release rate of BSA-FITC from skin. These release profiles of BSA-FITC in binary mixtures were expressed by Fick-s law of diffusion. These results suggest the utilization of hollow microneedle to enhance transdermal delivery of protein and provide useful information for designing an effective hollow microneedle system.

Keywords: Hydrophilic macromolecules, Microneedles, Propylene glycol, Transdermal drug delivery

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Authors: Roxana Mendoza Huaitalla, Eva Gallmann, Kun Zheng, Xuejun Liu, Eberhard Hartung

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Porcine production in China represents approximately the 50% of the worldwide pig production. Information about pig husbandry characteristics in China and manure properties from sows to fatteners in intensive pig farms are not broadly available for scientific studies as it is a time consuming, expensive task and highly inaccessible. This study provides a report about solid pig manures (28% dry matter) in a commercial pig farm located in the peri-urban area of Beijing as well as a general overview of the current pig husbandry techniques including pig breeds, feeds, diseases, housing as well as pig manure and wastewater disposal. The main results are intended to serve as a literature source for young scientists in order to understand the main composition of pig manures as well as to identify the husbandry techniques applied in an intensive pig farm in Beijing.

Keywords: China, heavy metals, intensive pig farming, manure, nutrients, pig growing stages.

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Authors: Wai Prathumpai, Pranee Rachtawee, Sutamat Khajeeram, Pariya Na Nakorn

Abstract:

The  β-glucan produced by Ophiocordyceps dipterigena BCC 2073 is a (1, 3)-β-D-glucan with highly branching O-6-linkedside chains that is resistant to acid hydrolysis (by hydrochloric acid and porcine pancreatic alpha-amylase). This β-glucan can be utilized as a prebiotic due to its advantageous structural and biological properties. The effects of using this β-glucan as the sole carbon source for the in vitro growth of two probiotic bacteria (L. acidophilus BCC 13938 and B. animalis ATCC 25527) were investigated. Compared with the effect of using 1% glucose or fructo-oligosaccharide (FOS) as the sole carbon source, using 1% β-glucan for this purpose showed that this prebiotic supported and stimulated the growth of both types of probiotic bacteria and induced them to produce the highest levels of metabolites during their growth. The highest levels of lactic and acetic acid, 10.04 g·L^-1 and 2.82 g·L^-1, respectively, were observed at 2 h of cultivation using glucose as the sole carbon source. Furthermore, the fermentation broth obtained using 1% β-glucan as the sole carbon source had greater antibacterial activity against selected pathogenic bacteria (B. subtilis TISTR 008, E. coli TISTR 780, and S. typhimurium TISTR 292) than did the broths prepared using glucose or FOS as the sole carbon source. The fermentation broth obtained by growing L. acidophilus BCC 13938 in the presence of β-glucan inhibited the growth of B. subtilis TISTR 008 by more than 70% and inhibited the growth of both S. typhimurium TISTR 292 and E. coli TISTR 780 by more than 90%. In conclusion, O. dipterigena BCC 2073 is a potential source of a β-glucan prebiotic that could be used for commercial production in the near future.

Keywords: β-glucan, Ophiocordyceps dipterigena, prebiotic, probiotic, antimicrobial.

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Authors: Jienny Lee, In-Soo Cho, Sang-Ho Cha

Abstract:

Adult mesenchymal stem cells (MSCs) have been investigated using preclinical approaches for tissue regeneration. Porcine MSCs (pMSCs) are capable of growing and attaching to plastic with a fibroblast-like morphology and then differentiating into bone, adipose, and cartilage tissues in vitro. This study was conducted to investigate the proliferating abilities, differentiation potentials, and multipotency of miniature pig adipose tissue-derived MSCs (mpAD-MSCs) with or without long-term cryopreservation, considering that cryostorage has the potential for use in clinical applications. After confirming the characteristics of the mpAD-MSCs, we examined the effect of long-term cryopreservation (> 2 years) on expression of cell surface markers (CD34, CD90 and CD105), proliferating abilities (cumulative population doubling level, doubling time, colony-forming unit, and MTT assay) and differentiation potentials into mesodermal cell lineages. As a result, the expression of cell surface markers is similar between thawed and fresh mpAD-MSCs. However, long-term cryopreservation significantly lowered the differentiation potentials (adipogenic, chondrogenic, and osteogenic) of mpAD-MSCs. When compared with fresh mpAD-MSCs, thawed mpAD-MSCs exhibited lower expression of mesodermal cell lineage-related genes such as peroxisome proliferator-activated receptor-g2, lipoprotein lipase, collagen Type II alpha 1, osteonectin, and osteocalcin. Interestingly, long-term cryostoraged mpAD-MSCs exhibited significantly higher cell viability than the fresh mpAD-MSCs. Long-term cryopreservation induced a 30% increase in the cell viability of mpAD-MSCs when compared with the fresh mpAD-MSCs at 5 days after thawing. However, long-term cryopreservation significantly lowered expression of stemness markers such as Oct3/4, Sox2, and Nanog. Furthermore, long-term cryopreservation negatively affected expression of senescence-associated genes such as telomerase reverse transcriptase and heat shock protein 90 of mpAD-MSCs when compared with the fresh mpAD-MSCs. The results from this study might be important for the successful application of MSCs in clinical trials after long-term cryopreservation.

Keywords: Mesenchymal stem cells, Cryopreservation, Stemness, Senescence.

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Authors: Eun-Geun Kim, Won-Pil Park, Dae-Gon Woo, Chang-Yong Ko, Yong-Heum Lee, Dohyung Lim, Tae-Min Shin, Han-Sung Kim, Gyoun-Jung Lee

Abstract:

Functional gastrointestinal disorders affect millions of people spread all age regardless of race and sex. There are, however, rare diagnostic methods for the functional gastrointestinal disorders because functional disorders show no evidence of organic and physical causes. Our research group identified recently that the gastrointestinal tract well in the patients with the functional gastrointestinal disorders becomes more rigid than healthy people when palpating the abdominal regions overlaying the gastrointestinal tract. Aim of this study is, therefore, to develop a diagnostic system for the functional gastrointestinal disorders based on ultrasound technique, which can quantify the characteristic above related to the rigidity of the gastrointestinal tract well. Ultrasound system was designed. The system consisted of transmitter, ultrasonic transducer, receiver, TGC, and CPLD, and verified via a phantom test. For the phantom test, ten soft-tissue specimens were harvested from porcine. Five of them were then treated chemically to mimic a rigid condition of gastrointestinal tract well, which was induced by functional gastrointestinal disorders. Additionally, the specimens were tested mechanically to identify if the mimic was reasonable. The customized ultrasound system was finally verified through application to human subjects with/without functional gastrointestinal disorders (Normal and Patient Groups). It was identified from the mechanical test that the chemically treated specimens were more rigid than normal specimen. This finding was favorably compared with the result obtained from the phantom test. The phantom test also showed that ultrasound system well described the specimen geometric characteristics and detected an alteration in the specimens. The maximum amplitude of the ultrasonic reflective signal in the rigid specimens (0.2±0.1Vp-p) at the interface between the fat and muscle layers was explicitly higher than that in the normal specimens (0.1±0.0Vp-p). Clinical tests using our customized ultrasound system for human subject showed that the maximum amplitudes of the ultrasonic reflective signals near to the gastrointestinal tract well for the patient group (2.6±0.3Vp-p) were generally higher than those in normal group (0.1±0.2Vp-p). Here, maximum reflective signals was appeared at 20mm depth approximately from abdominal skin for all human subjects, corresponding to the location of the boundary layer close to gastrointestinal tract well. These results suggest that newly designed diagnostic system based on ultrasound technique may diagnose enough the functional gastrointestinal disorders.

Keywords: Functional Gastrointestinal Disorders, DiagnosticSystem, Phantom Test, Ultrasound System.

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