• Development and Reproduction
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• v.2 no.2
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• pp.165-171
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• 1998

p62 is a novel cytoplsmic protein that binds to SH2 domain of p56$^{lck}$, lymphocyte-specific protein tyrosine kinase, and the expression of p62 was observed in most tissues. In addition p62 interacts with various proteins including ubiquitin and atypical PKC isoform, indicating its diverse biological role in different tissues. However, little is known about functional connection between p62 and its binding proteins. In the present study, a novel cellular protein, p62 has been shown to bind to 14-3-3 $\tau$ isoform that is specific for T cells. Moreover, overexpression of p62 in T cells caused to delay onset of UV-induced apoptosis characterized by DNA fragmentation and breakdown of poly (ADP-ribose) polymerase (PARP). Lately, 14-3-3 proteins have been shown to mediate survival signal via interacting proapoptotic Bad protein in the Iymphocyte. These results suggested the presence of p62-mediated regulatory mechanism during apoptosis in T cells, in which activation-induced apoptotic signal could be interfered by p62 and 14-3-3 protein.n.

• Applied Microscopy
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• v.32 no.4
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• pp.329-337
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• 2002

The morphology and cellular characteristics of adventitious roots, viz aerial roots, in the epiphytic American Ivy were examined to reveal structural changes of the aerial root upon surface attachment. Immature aerial roots were composed of parenchyma cells with dense cytoplasm containing plastids, however, the upper and lower epidermis were not distinguished. At early development, electron-dense substances (EDS) were constituents of much of the aerial root tissue, but the distribution of EDS varied within the tissue. The deposits appeared most concentrated in the superficial cell layers, with lesser amounts in cell layers closer to the cortex. Electron micrographs revealed that EDS deposits were always found in the vacuole, and were mainly associated with the tonoplast. While most of them occurred in the vacuole as small spherical deposits adjacent to the tonoplast, some deposits were oddly shaped or larger in size. Many of the vacuoles eventually filled with EDS, but the EDS content in those vacuoles decreased substantially after initial attachment to the surface. When the vacuoles became almost empty, cells near the epidermis already exhibited irregularity in outline. Subsequent breakdown of cellular components took place in the cells while they were still attached to the surface. This study suggests the potential role of EDS as substances involved in the surface attachment of the plant, however, further studies must be conducted to reveal the nature of EDS and the effects of EDS storage within these vacuoles.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.179-188
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• 2017

White-rot basidiomycetes are the organisms that decompose lignin most efficiently, and Trametes villosa is a promising species for ligninolytic enzyme production. There are several publications on T. villosa applications for lignin degradation regarding the expression and secretion of laccase and manganese peroxidase (MnP) but no reports on the identification and characterization of lignin peroxidase (LiP), a relevant enzyme for the efficient breakdown of lignin. The object of this study was to identify and partially characterize, for the first time, gDNA, mRNA, and the corresponding lignin peroxidase (TvLiP) protein from T. villosa strain CCMB561 from the Brazilian semiarid region. The presence of ligninolytic enzymes produced by this strain grown in inducer media was qualitatively and quantitatively analyzed by spectrophotometry, qPCR, and dye fading using Remazol Brilliant Blue R. The spectrophotometric analysis showed that LiP activity was higher than that of MnP. The greatest LiP expression as measured by qPCR occurred on the $7^{th}$ day, and the ABSA medium (agar, sugarcane bagasse, and ammonium sulfate) was the best that favored LiP expression. The amplification of the TvLiP gene median region covering approximately 50% of the T. versicolor LPGIV gene (87% identity); the presence of Trp199, Leu115, Asp193, Trp199, and Ala203 in the translated amplicon of the T. villosa mRNA; and the close phylogenetic relationship between TvLiP and T. versicolor LiP all indicate that the target enzyme is a lignin peroxidase. Therefore, T. villosa CCMB561 has great potential for use as a LiP, MnP, and Lac producer for industrial applications.

• Journal of Microbiology and Biotechnology
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• v.28 no.9
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• pp.1547-1553
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• 2018

Hyaluronidases are a family of enzymes that catalyse the breakdown of hyaluronic acid, which is abundant in the extracellular matrix and cumulus oocyte complex. To investigate the activity of recombinant bovine sperm hyaluronidase 1 (SPAM1) and determine the effect of the Asn-X-Ser/Thr motif on its activity, the bovine SPAM1 open reading frame was cloned into the mammalian expression vector pCXN2 and then transfected to the HEK293 cell line. Expression of recombinant bovine hyaluronidase was estimated using a hyaluronidase activity assay with gel electrophoresis. Recombinant hyaluronidase could resolve highly polymeric hyaluronic acid and also caused dispersal of the cumulus cell layer. Comparative analysis with respect to enzyme activity was carried out for the glycosylated and deglycosylated bovine sperm hyaluronidase by N-glycosidase F treatment. Finally, mutagenesis analysis revealed that among the five potential N-linked glycosylation sites, only three contributed to significant inhibition of hyaluronic activity. Recombinant bovine SPAM1 has hyaluronan degradation and cumulus oocyte complex dispersion ability, and the N-linked oligosaccharides are important for enzyme activity, providing a foundation for the commercialization of hyaluronidase.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.2
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• pp.174-183
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• 2001

During the rapid phase of gonadal development of the freshwater teleost, the catfish (Silurus asotus), the influence of hCG upon the inducement of final oocyte maturation and spawning was investigated electrophoretically and ultrastructurally. The electrophoretic patterns obtained were different in the presence and absence of some of the major or minor zones, because of the hormone level in catfish. The vitellogenin of hormone-treated fish was stained more intensively than that of sham-treated fish. These proteins showed some minor or main bands of egg extracts which migrated at positions corresponding to molecular weights of approximately 90,000. However, the thickness of electrophoretic band in molecular weight for hCG-treated fish was slightly lower than that for saline control. It seemed the plasma protein with molecular weight of approximately 45,000 in hCG-treated fish disappeared. In contrast to the control fish, the ovaries in the catfish treated with hCG shows a marked ultrastructural change under the electron microscope. No dilated profiles were seen in the granulosa cells of the mature oocyte before ovulation. After germinal vesicle breakdown (GVBD), the zona radiata interna (ZRI) becomes more compact, and there is a loss of all the processes from the pore canals. There is a wide space between the vitelline membrane and zona radiata. Also, during final maturation, the microvillar processes from the oocyte are seen no longer to penetrate deeply into the extracellular spaces of the overlying granulosa cells, and the reticulate patterns of the zona radiata interna becomes occluded, giving the zona radiata a more solid appearance. It has been possible to initiate 100% oocyte maturation in yolk granules and follicles in vivo by treatment with hCG and a high water temperature ($27^{\circ}C$). In hCG-treated fish, the percentages of successful artificial fertilization and hatching were maximal at 15 h after a single injection. It seems clear that a long acting preparation containing hCG can be successfully used in prespawning fish to advance the final events of gonadal maturation and initiate spawning. Further studies are necessary to evaluate the potential of hCG to either stimulate or inhibit the reproductive development of fish at other stages of the seasonal reproductive cycle.

• Archives of Reconstructive Microsurgery
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• v.11 no.2
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• pp.153-161
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• 2002

Purpose : Plantar surfaces, calcaneal area, and region of Achilles insertion, which are extremely related with weight-bearing area and shoes application, must be reconstructed with glabrous and strong fibrous skin. Numerous methods of reconstructing defects of these regions have been advocated, but the transfer of similar local tissue as a cutaneous flap with preservation of sensory potential would best serve the functional needs of the weight-bearing and non-weight-bearing surfaces of this region. Therefore it is recommended to use the limited skin of medial surface of foot that is similar to plantar region and non-weight-bearing area. In this paper we performed the medial plantar flap transfered as a fasciocutaneous island as one alterative for moderate-sized defects of the plantar forefoot, plantar heel, and area around the ankle in 25 cases and report the result, availability and problem of medial plantar flap. Materials and methods : We performed proximally based medial plantar flap in 22 cases and reverse flow island flap in 3 cases. Average age was $36.5(4{\sim}70)$ years and female was 3 cases. The causes of soft tissue defect were crushing injury on foot 4 cases, small bony exposure at lower leg 1 case, posterior heel defect with exposure of calcaneus 8 cases, severe sore at heel 2 cases, skin necrosis after trauma on posterior foot 4 cases, and defect on insertion area of Achilles tendon 6cases. Average follow up duration was 1.8(7 months-9.5 years) years. Results: Medial plantar flaps was successful in 22 patients. 18 patients preserved cutaneous branches of medial plantar nerve had sensation on transfered flap but diminished sensation or dysesthesia. At the follow up, we found there were no skin ulceration, recurrence of defect or skin breakdown in all 18 patients. But there was one case which occurred skin ulceration postoperatively among another 4 cases not contained medial plantar nerve. At the last follow up, all patients complained diminished sensation and paresthesia at medial plantar area distally to donor site, expecially with 4 patients having severe pain and discomfort during long-time walking. Conclusion : Medial plantar island flap based on medial plantar neurovascualr pedicle have low failure rate with strong fibrous skin and preserve sensibility of flap, so that it is useful method to reconstruct the skin and soft tissue defect of foot. But it should be emphasized that there are some complications such like pain and paresthesia by neuropraxia or injury of medial plantar nerve at more distal area than donor site. We may consider that medial plantar flap have limited flap size and small arc of rotation, and require skin graft closure of the donor defect and must chose this flap deliberately.

• Journal of Bio-Environment Control
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• v.12 no.3
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• pp.166-172
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• 2003

Rockwool slabs used for hydroponic cultures are expected to have potential to be reused after breakdown as a growing medium component for hydroponic culture of vegetables and cut flowers, pot plants, or plug plants. This study was conducted to test the feasibility of using particles of used rockwool slabs (PURS) mixed with woodchips for growing plug seedlings of ‘Nokkufang’ pepper. In the first experiment growth of pepper seedlings was tested in media of steam sterilized PURS mixed with chestnut woodchips (CW), which was weatherized for six months and screened through 2.8 mm or 5.6 mm sieves, at 100:0, 75:25, 50:50, 28:75 or 0:100 (%, v/v). In the second experiment growth of pepper seedlings was tested in 20 different media containing steam sterilized PURS mixed with 4 mm chestnut woodchips (CW), 4 mm pinc woodchips (PW), coir, peatmoss, or perlite. In the first experiment, there were no significant growth differences as affectcd by particle size of CW, while the overall best growth was achieved il1 the control and l00% PURS. In the second experiment, seedling growth was enhanced as compared to the control in 100% PURS or PURS mixed with peatmoss, coir, or perlite. In contrast, growth was suppressed as the proportion of peatmoss or coir mixed with PURS decreased and the proportion of CW increased. Growth was better in the PURS + peatmoss than in the 100% PURS or PURS + coir, and when woodchips were mixed with coir than peatmoss.

• Korean Journal of Food Science and Technology
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• v.31 no.2
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• pp.410-415
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• 1999

Yakju was sterilized with high-voltage pulses of short time of a continuous pulsed electric field (PEF) system. The initial microbial counts of Yakju were $2.2{\times}10^{5}$ CFU/mL for total aerobes. The pH, acidity and electric conductivity of Yakju were 3.82, 0.37% and 1.24 mS/cm, respectively. Yakju was treated with exponential-wave formed electric pulses of 100 Hz for $0{\sim}4000{\mu}s$ under the field strength of $20{\sim}35\;kV/cm$. The lethal effect of electric fields on microorganisms was resulted from the breakdown of the cell membrane induced by the transmembrane electric potential. The critical values of the external field for the sterilization were 16.0 kV/cm for total aerobes. Logarithmic survival rates decreased linearly at low electric field strength, but curvilinearly at high electric field strength with treatment time. The sterilization of Yakju was more largely affected by the electric field strength than by the treatment time. Any changes in pH, acidity, and the growth of microorganisms were not found in the PEF treated Yakju during the storage at both $4^{\circ}C\;and\;30^{\circ}C$.

• Journal of Animal Science and Technology
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• v.46 no.5
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• pp.871-878
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• 2004

Pulsed electric fields(PEF) technology is one of the latest nonthermal methods of food processing for obtaining safe and minimally processed foods. This technology can be effectively explored for obtaining safe food with minimum effect on nutritional, flavor, rheological and sensory qualities of food products. The process involves the application of high voltage(typically 20 ${\sim}$ 80 kv/cm) to foods placed between two electrodes. The mode of inactivation of microorganism; by PEP processing has been postulated in term; of electric breakdown and electroporation. The extent of destruction of microorganisms in PEF processing depends mainly on the electric field strength of the pulses and treatment time. For each cell types, a specific critical electric field strength and specific critical treatment time are required depending on the cell characteristics and the type and strength of the medium where they have been present. The effect also depends on the types of microorganisms and their phase of growth. A careful combination of processing parameters has to be selected for effective processing. The potential applications of PEF technology are numerous ranging from biotechnology to food preservation. With respect to food processing, it has already been established that, the technology is non-thermal in nature, economical and energy efficient, besides providing minimally processed foods. This article gives a brief overview of this technology for food processing applications.

• Journal of agriculture & life science
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• v.44 no.2
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• pp.57-66
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• 2010

The antioxidant and neuronal cell protective effects of water extract from purple sweet potato were investigated. The total phenolics and monomeric anthocyanin contents of purple sweet potato extract were 44.25 mg/g and 2,394 mg/L, respectively. The antioxidant activities of purple sweet potato extract were evaluated using various antioxidant tests, including 1,1-diphenyl- 2-picrylhydrazyl (DPPH), 2,2'-azino- bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, ferric reducing/antioxidant power (FRAP) and reducing power. In these assays, the extract of purple sweet potato presented significant radical scavenging activities, FRAP, and reducing power in a dose-dependent manner. MTT {3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl- tetrazoliumbromide} reduction assay showed significantly increase in cell viability when PC12 cells were pretreated with purple sweet potato extract. Because oxidative stress is also known to increase neuronal cell membrane breakdown, we further investigated by lactate dehydrogenase (LDH) and neutral red uptake assay. Purple sweet potato extract inhibited oxidative stress-induced membrane damage in neuronal cells. Therefore, these data results demonstrated that water extract of purple sweet potato have antioxidant activity and neuronal cell protective effect thus it has great potential as a natural source for human health.