• Journal of Embryo Transfer
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• v.15 no.1
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• pp.103-108
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• 2000

The object of this study was to find simple and effective methods for the speculation of vitality and scorsome status of bovine spermatozoa. The eosin-nigrosin staining, trypan blue staining, and naphthol yellow S-erythrosin B staining was ofter used for the speculation of vitality and/or acrosome status of bovine spermatozoa, respectively. This study has shown that the combined trypan blue-naphthol yellow S-erythrosin B staining is more accurate and effective for the examination of acrosome status and vitality of bovine spermatozoa.

• Journal of the korean veterinary medical association
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• v.16 no.1
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• pp.25-28
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• 1980

The studies were undertaken with the objective to compare six staining techniques, nigrosineosin, eosin-nigrosin, trypan blue, Congo red-nigrosin, fast green-erythrocin and fast green-eosin and select one of these staining techniques to measure live-dead

• Journal of Fisheries and Marine Sciences Education
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• v.28 no.6
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• pp.1822-1827
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• 2016

Consumption of Paralichthys olivaceus in raw fish have been reported as the cause of outbreaks of food-born illness, and Kudoa septempunctata in muscle of Paralichtys olivaceus was suggested with the causative agent. For this reason, distinguish of vital and dead spore is important to study survivability of Kudoa septempunctata in human intestinal condition and in vitro inactivation of Kudoa septempunctata. In this reports, we suggest NR & MB (Neutral red and Methylene blue) staining method that is easier and simpler than the previously described HO & PI (Hoechst33342 and Propidium iodide) method according to a experimental condition.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.429-433
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• 2000

Giardia lamblia, a human pathogen causing outbreaks of diarrhea, recently became a focus of great concerns in the fields of both medical and environmental microbioloty. To develop the experimental tools to study giardiasis, encystation, one of the major processes in its life cycle, was reconstituted by inducing an axenic culture of a flagellated form of G. lamblia into a cyst from under high concentration of bile and alkaline pH condition. The successful induction was confirmed by Northern analysis of resulting increased expression of the CWPl gene encoding the cyst wall protein 1. An examination of the encystation process with SEM (scanning electron microscopy) and TEM (transmission electron microscopy) revealed that the trophozoite, a flagellate with a bilateral symmetry, was transformed to a cyst form with an oval-shape and defined filamentous wall. The encystation was found to cause a disappearance of the flagella and an invagination of the adhesive disc. An extensive formation of rER (rough endoplasmic reticulum) was observed after 24h of induction, indication an active synthesis and export of proteins during this process. The vital staining of the invitro-induced systs showed that most cysts maintained their viability.

• Journal of Veterinary Clinics
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• v.22 no.3
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• pp.228-232
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• 2005

Damaged spermatozoa are supposed to be trapped in glass wool. In the respect of this, two glass wool filtration spermatozoa groups (0.5 cm, 1 cm depth) were compared with control group to assay sperm motility, HOS values, and vital rate by CFDA/PI staining method following glass wool filtration. The motility of canine sperm extended with PBS+PVP after glass wool filtration was lower in both filtrated groups than that of the control group (p<0.01) and the same significant difference was also shown in canine semen extended with Tris buffer (p<0.01). The motility of canine sperm diluted with PBS+PVP was higher than that diluted with Tris buffer in the same experimental groups (p<0.05). The motility of control group was not significantly decreased until 2 hours immediately after extending, however, the motility of both glass wool filtrated spermatozoa were significantly decreased as time passed until 2 hours after filtration (p<0.01). At each time for assay (immediately, 30 min, 2 hours after filtration), the motility of canine sperm of control group was higher than the filtrated groups (p<0.05), whereas the motility of 0.5 cm depth group was higher than 1 cm depth group at the immediate time after filtration (p<0.05), 30 minutes later (p<0.05) with no difference at 2 hours. No difference was shown among the experimental groups in HOS values of canine sperm after glass wool filtration. The vital rate assayed by CFDA/PI staining of both filter groups was higher than the control group (p<0.05).

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.3
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• pp.975-978
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• 2012

P63 is a gene product required in cell cycle regulation which plays vital roles in tumor differentiation. Aims of the present study were to assess the frequency, pattern, sensitivity and specificity of two p63 protein clones P63 4A4 and P63 4A4+Y4A3 in squamous cell carcinomas (SCCs). Thirty cases of head and neck region SCC diagnosed on the basis of H&E staining were examined along with 60 cases of head and neck region biopsies other than squamous cell carcinoma, negative on H&E staining, were taken as control. Immunostaining was performed on slides according to the Thermo Scientific UltraVision LP detection System. P63 4A4+Y4A3 clone is more sensitive 96.6% in comparison to 86% in P63 4A4 with having greater NPV of 98.3%. The results signify the importance of P63 4A4+Y4A3 marker over the old markers and may be used as a confirmatory marker of squamous cell carcinoma.

• Restorative Dentistry and Endodontics
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• v.3 no.1
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• pp.17-21
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• 1977

After a vital pulpotomy in dogs' teeth, the responses of the remaining pulp tissue under calcium hydroxide and formocresol were studied histologically. The class I and V cavities were prepared on the teeth and the pulp was amputated. Calcium hydroxide and formocresol were placed over the amputated tissue and the cavities were sealed with zine oxide eugenol cement and zinc phosphate cement. Animals. were sacrifice after 1, 2, and 3 weeks following the operation. The teeth were decalcfied, sectioned and stained with hematoxylin and eosin. Microscopic examination reveals as follows; 1. Healing of the pulp at the amputation site did not occur in the pulps treated with formocresol. 2. At one week, a thin layer of darker staining tissues just below the necrotic zone was presented in the pulps treated with formocresol. In this stage the tissues beneath the darker staining layer were normal. 3. At two weeks, the cells of the palest staining layer were showed indistinct nucleus which suggested the karyolysis and the karyorrhexis in the pulps treated with formocoresol. As reached to the middle third of the pulp, the odontoblasts were scarcely evident or missed in this stage. 4. At three weeks, the necrotic zone was reached to the middle third of the pulp canal. The cells beneath the zone showed massive infiltration of inflammatory cells in the pulps treated with formocresol. 5. Dentin bridge in the control group was deposited below the necrotic zone from the two. weeks later. 6. Normal tissues were observed ill the apical third of all. the dental pulps in all case of calcium hydroxide and formocresol.

• Journal of Korean Neurosurgical Society
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• v.30 no.6
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• pp.683-687
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• 2001

Objective : Angiogenesis, the proliferation of capillary endothelial cells, is a vital component in the development, progression, and metastasis of many human tumors. Vascular endothelial growth factor(VEGF) is an endothelial cell-specific mitogen and induces angiogenesis and vascular permeability. The features of glioblastoma, distinct from low grade astrocytomas, are the presence of necroses and vascular endothelial proliferation. In this study, we investigated VEGF expression in the different grades of astrocytomas and determined whether VEGF expression correlates with development of glioblastoma and progression of astrocytomas. Patients and Methods : Forty seven patients with astrocytic tumors(24 males and 23 females), aged 3 to 65 years, were evaluated. Immunohistochemical staining was carried out using labelled streptavidin biotin method and primary antibody was a antirabbit polyclonal Ab against N-terminus region of VEGF165(Oncogene research product, MA, USA). Immunoreactivity(IR) was classified into no IR(absent or a trace of stain), moderate IR and intense IR by level of staining amount and intensity. Results : Six pilocytic astrocytomas showed 3 no IR and 3 moderate IR, 10 astrocytomas showed 2 no IR, 6 moderate IR and 2 intense IR, 12 anaplastic astrocytomas showed I no IR, 7 moderate IR and 4 intense IR and 19 glioblastomas showed 1 no IR, 11 moderate IR and 7 intense IR. Immunoreactivity was significantly different between low and high grade of tumors but there was no significant difference between anaplastic astrocytomas and glioblastomas. Gemistocytic tumor cells represented the predominent VEGF-immunoreactive cell types, as compared with compactly-arranged small tumor cells. In glioblastomas VEGF IR was observed in both perinecrotic and vital tumor areas. Conclusion : VEGF seems to be a important angiogenic factor in anaplastic astrocytomas and glioblastomas and VEGF expression may contribute to neovascularization of human astrocytomas.

• Advances in nano research
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• v.13 no.6
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• pp.575-585
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• 2022

Due to its biofilm formation and colonization of the osteocyte-lacuno canalicular network (OLCN), Staphylococcus aureus (S.aureus) implant-associated bone infection (SIABI) is difficult to cure thoroughly, and may occur recurrently subsequently after a long period dormant. It is essential to explore an alternative therapeutic strategy that can eradicate the pathogens in the infected foci. To address this, the polymethylmethacrylate (PMMA) bone cement and Fe3O4 nanoparticles compound cylinder were developed as implants based on their size and mechanical properties for the alternative magnetic field (AMF) induced thermal ablation, The PMMA mixed with optimized 2% Fe₃O₄ nanoparticles showed an excellent antibacterial efficacy in vitro. It was evaluated by the CFU, CT scan and histopathological staining on a rabbit 1-stage transtibial screw model. The results showed that on week 7, the CFU of infected soft tissue and implants, and the white blood cells (WBCs) of the PMMA+2% Fe₃O₄+AMF group decreased significantly from their controls (p<0.05). PMMA+2% Fe₃O₄+AMF group did not observe bone resorption, periosteal reaction, and infectious reactive bone formation by CT images. Further histopathological H&E and Gram Staining confirmed there was no obvious inflammatory cell infiltration, neither pathogens residue nor noticeably burn damage around the infected screw channel in the PMMA+2% Fe₃O₄+AMF group. Further investigation of nanoparticle distributions in bone marrow medullary and vital organs of heart, liver, spleen, lung, and kidney. There were no significantly extra Fe₃O₄ nanoparticles were observed in the medullary cavity and all vital organs either. In the current study, PMMA+2% Fe₃O₄+AMF shows promising therapeutic potential for SIABI by providing excellent mechanical support, and promising efficacy of eradicating the residual pathogenic bacteria in bone infected lesions.

• Restorative Dentistry and Endodontics
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• v.48 no.2
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• pp.21.1-21.15
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• 2023

Objectives: This study evaluated the effects of Biodentine (BD), Bio-C Repair (BCR), and mineral trioxide aggregate (MTA) plug on the fracture resistance of simulated immature teeth with replacement root resorption (RRR) and in vitro-induced osteoclastogenesis. Materials and Methods: Sixty bovine incisors simulating immature teeth and RRR were divided into 5 groups: BD and BCR groups, with samples completely filled with the respective materials; MTA group, which utilized a 3-mm apical MTA plug; RRR group, which received no root canal filling; and normal periodontal ligament (PL) group, which had no RRR and no root canal filling. All the teeth underwent cycling loading, and compression strength testing was performed using a universal testing machine. RAW 264.7 macrophages were treated with 1:16 extracts of BD, BCR, and MTA containing receptor activator of nuclear factor-kappa B ligand (RANKL) for 5 days. RANKL-induced osteoclast differentiation was assessed by staining with tartrate-resistant acid phosphatase. The fracture load and osteoclast number were analyzed using 1-way ANOVA and Tukey's test (α = 0.05). Results: No significant difference in fracture resistance was observed among the groups (p > 0.05). All materials similarly inhibited osteoclastogenesis (p > 0.05), except for BCR, which led to a lower percentage of osteoclasts than did MTA (p < 0.0001). Conclusions: The treatment options for non-vital immature teeth with RRR did not strengthen the teeth and promoted a similar resistance to fractures in all cases. BD, MTA, and BCR showed inhibitory effects on osteoclast differentiation, with BCR yielding improved results compared to the other materials.