• Asian-Australasian Journal of Animal Sciences
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• 제23권8호
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• pp.993-999
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• 2010

Serum lysozyme gene is one of the important genes influencing the immune system as its product can cause lysis of bacterial cell wall by cleaving the peptidoglycan layer. The present investigation on the serum lysozyme gene of Indian riverine buffalo was undertaken with the objectives to identify and characterize single nucleotide polymorphic patterns by PCR-SSCP method as well as to study the effect of different genotypes on serum lysozyme activity and somatic cell count. A total of 280 animals comprising four different famous bubaline breeds (Murrah, Mehsana, Surti and Bhadawari), spread over six different farms across the country were used for this study. A 276 bp (partial intron 2, complete exon 3 and partial intron 3) fragment of lysozyme gene was screened for polymorphism using the SSCP technique. Four genotypes namely AA, AB, BC and AC were observed, out of which BC genotype was found to be the most frequent. Among these three alleles, C allele (0.38) was most prevalent in these populations. Various SSCP allelic variants were cloned for sequencing and sequences were submitted to NCBI Genbank. From the alignment of the nucleotide sequences of various allelic variants, it was found that there were differences in 12 positions among the alleles, out of which maximum variation (at 8 places) was found in the intronic region. The allele A was closer to allele-C than allele-B. Allele B was phylogenetically equidistant from both of the other alleles. Mean lysozyme activity determined in serum samples of different animals of Murrah buffalo was $27.35{\pm}2.42\;{\mu}g$ per ml of serum, whereas the mean somatic cell count was $1.25{\pm}0.13{\times}10^5$ cells per ml of milk. The SSCP pattern-wise effects of various genotypes on lysozyme activity and SCC were analyzed. Although the mean values were apparently different in various genotypes, these differences were statistically non-significant. It can be concluded that the riverine buffaloes are sufficiently polymorphic with respect to serum lysozyme gene. The absence of AA genotype in Bhadawari breed of buffalo can be considered as a marker for breed characterization. The difference of four nucleotides in exon-3 indicates high selection pressure on the gene.

• 한국가축번식학회지
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• 제17권2호
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• pp.85-92
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• 1993

In the previous study, we observed that Purine has a time dependent effect in maintaining the oocytes in meiotic arrest, and human fetal cord serum(HFCS) and human mature follicular fluid(HMFF) reverse the GVBD suppressed by purines. And it was reported that purine has a harmful effect on the development of oocytes or embryos, when they were cultured for a long time, in vitro. Therefore this study was performed to investigate the effects of purine on extrusion rates of 1st pb and viability of oocytes cultured for a long time, in vitro. Immature oocytes(GV stage) were collected from ovaries of 25~28 day old ICR mice at 48 hrs after PMSG injection. Cumulus-enclosed and denuded oocytes collected were assigned randomly to one of several culture conditions. Some of the oocytes were cultured in 4mM hypoxanthine for 24hr, and the extrusion rates of 1st pb and viability of the oocytes were assessed at every 12 hrs. In the viability, the oocytes showed granulation, pigmentation of cytoplasm or lysis of 1st pb extruded were regarded as degenerating oocytes. Also some of the oocytes were cultured in hypoxanthine for 12 hrs then the resulting oocytes were transferred to hypoxanthine-free medium and cultured for 12 hrs to determine whether the inhibitory effect of hypoxanthine on the 1st pb extrusion was reversible. The rest of the oocytes were cultured in medium containing hypoxanthine and adenosine for 18 hrs to compare the 1st pb extrusion be attendant upon hte concentration of HFCS or HMFF supplemented. Hypoxanthine suppressed the extrusion of 1st pb and viability of the oocytes significantly, when they were cultured for more than 12 hrs and the harmful effect of hypoxanthine was showed in denuded oocytes, prominently. The suppressive effect of hypoxanthine was reversed by just removal of the hypoxanthine from the cultrue medium. Also there was no difference in reverse the pb extrusion rate suppressed between HFCS and HMFF. The extrusion rate of 1st pb in medium containing adenosine and hypoxanthine was increased in line with the concentration of HFCS or HMFF supplemented. Hypoxanthine suppressed the extrusion of 1st pb and viability of the oocytes significantly, when they were cultured for more than 12 hrs and the harmful effect of hypoxanthine was showed in denuded oocytes, prominetly. The suppressive effect of hypoxanthine was reversed by just removal of the hypoxanthine fromthe culture medium. Also there was no difference in reverse the pb extrusion rate suppressed between HFCS and HMFF. The extrusion rate of 1st pb in medium containing adenosine and hypoxanthine was increased in line with the concentration of HFCS or HMFF supplemented.

• Asian Pacific Journal of Cancer Prevention
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• 제15권3호
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• pp.1219-1225
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• 2014

Epigallocatechin-3-gallate (EGCG) is the most abundant polyphenol molecule from green tea and is known to exhibit antioxidative as well as tumor suppressing activity. In order to examine EGCG tumor invasion and suppressing activity against adult T-cell leukemia (ATL), two HTLV-1 positive leukemia cells (HuT-102 and C91-PL) were treated with non-cytotoxic concentrations of EGCG for 2 and 4 days. Proliferation was significantly inhibited by 100 ${\mu}M$ at 4 days, with low cell lysis or cytotoxicity. HTLV-1 oncoprotein (Tax) expression in HuT-102 and C91-PL cells was inhibited by 25 ${\mu}M$ and 125 ${\mu}M$ respectively. The same concentrations of EGCG inhibited NF-kB nuclearization and stimulation of matrix metalloproteinase-9 (MMP-9) expression in both cell lines. These results indicate that EGCG can inhibit proliferation and reduce the invasive potential of HTLV-1-positive leukemia cells. It apparently exerted its effects by suppressing Tax expression, manifested by inhibiting the activation of NF-kB pathway and induction of MMP-9 transcription in HTLV-1 positive cells.

• 대한약침학회지
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• 제11권1호
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• pp.71-82
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• 2008

Objectives : The purpose of this study is to investigate the effects of Spirodelae Herba pharmacopuncture(SHP) on the adipogenesis in 3T3 cells, lipolysis in rat epididymal adipocytes and histological changes in porcine adipose tissue. Methods : Inhibition of preadipocyte differentiation and/or stimulation of lipolysis play important roles in reducing obesity. 3T3 preadipocytes were differentiated with adipogenic reagents by incubating for 3 days in the absence or presence of SHP ranging from 0.01 to $1.0mg/m{\ell}$. The effect of SHP on adipogenesis was examined by measuring glycerol-3-phosphate dehydrogenase(GPDH) activity and by Oil Red O staining. Mature adipocytes from rat epididymal fat pad was incubated with SHP ranging from 0.01 to $1.0mg/m{\ell}$ for 3 days. The effect of SHP on lipolysis was examined by measuring free glycerol released. Fat tissue from porcine skin was injected with SHP ranging from 0.1 to $10.0mg/m{\ell}$ to examine the effect of SHP onhistological changes under light microscopy. Results : Following results were obtained from the preadipocyte proliferation and lipolysis adipocyte and histologic investigation of fat tissue 1. SHP showed the effect of decreased preadipocyte proliferation on the high dosage($1mg/m{\ell}$). 2. SHP showed the effect of decreased the activity of glycerol-3-phosphate dehydrogenase (GPDH) on the high dosage($1mg/m{\ell}$). 3. Investigated the changes in lipolysis of differentiated adipocyte after treated SHP, we knew that these pharmacopuncture showed increasing the effect of lipolysis in all concentration significantly. 4. Investigated the histological changes in porcine fat tissue after treated SHP, we knew that these pharmacopuncture showed significant activity to the lysis of extensive cell membranes on high dosage($10.0mg/m{\ell}$). Conclusions : These results suggest that SHP efficiently induces diminishing proliferation of preadipocyte and lipolysis in adipose tissue.

• 대한약침학회지
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• 제11권1호
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• pp.163-176
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• 2008

Objective : The purpose of this study is to investigate the effects of Bangpungtongsung-san extracts on the preadipocytes proliferation, of 3T3-L1 cell line. lipolysis of adipocytes in rat's epididymis and localized fat accumulation of porcine by extraction methods(alcohol and water). Methods : Diminish 3T3-L1 proliferation and lipogenesis do primary role to reduce obesity. So, 3T3-L1 preadipocyte and adipocytes were performed on cell cultures, and using Sprague-Dawley rats for the lipogenesis, and treated with 0.01-$1mg/m{\ell}$ Bangpungtongsung-san Extracts depend on concentrations. Porcine skin including fat tissue after treated Bangpungtongsung-san Extracts by means of the dosage dependent variation are investigated the histologic changes after injection of these extracts. Results : Following results were obtained from the 3T3-L1 preadipocyte proliferation and lipolysis of adipocyte in rats and histologic investigation of fat tissue. 1. Bangpungtongsung-san extracts were showed the effect of decreased preadipocyte proliferation on the high dosage($1.0mg/m{\ell}$). 2. Bangpungtongsung-san extracts were showed the effect of decreased the activity of glycerol-3-phosphate dehydrogenase(GPDH) on the high dosage($1.0mg/m{\ell}$) and Specially, alcohol extract of Bangpungtongsung-san was clear as time goes by high concentration. 3. Bangpungtongsung-san extracts were showed tries to compare the effect of lipolysis, alcohol extract of Bangpungtongsung-san on the high dosage($1.0mg/m{\ell}$) was observed the effect is higher than water extract. 4. Investigated the histological changes in porcine fat tissue after treated Bangpungtongsung-san extracts, we knew that water extract of Bangpungtongsung-san was showed the effect of lipolysis on the high dosage($10.0mg/m{\ell}$) and alcohol extract of Bangpungtongsung-san was showed significant activity to the lysis of cell membranes in all concentration. Conclusion : These results suggest that Bangpungtongsung-san efficiently induces diminish proliferation of preadipocyte and lipolysis in adipose tissue.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제31권6호
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• pp.544-549
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• 2009

Failure to use effective methods of reduction, fixation and immobilization may lead to osteomyelitis with the exposed necrotic bone, as the overzealous use of transosseous wires & plates that devascularizes bone segments in the compound comminuted fractures of mandible. Once osteomyelitis secondary to fractures has become established, intermaxillary fixation should be instituted as early as possible. Fixation enhances patient comfort and hinders ingress of microorganisms and debris by movement of bone fragments. Teeth and foreign materials that are in the line of fracture should be removed and initial debridement performed at the earliest possible time. Grossly necrotic bone should be excised as early as possible ; no attempt should be made to create soft tissue flaps to achieve closure over exposed bone. The key to treatment of chronic osteomyelitis of the mandible is adequate and prolonged soft tissue drainage. If good soft tissue drainage is provided over a long period, sequestration of infected bone followed by regeneration or fibrous tissue replacement will occur so that appearance and function are not seriously altered. Localization and sequestration of infected mandible are far better performed by natural mechanism of homeostasis than by cutting across involved bone with a cosmetic or functional defect. As natural host defenses and conservative therapy begin to be effective, the process may become chronic, inflammation regresses, granulation tissue is formed, and new blood vessels cause lysis of bone, thus separating fragments of necrotic bone(sequestra) from viable bone. The sequestra may be isolated by a bed of granulation tissue, encased in a sheath of new bone(involucrum), and removed easily with pincettes. This is a case report of the long-term conservative drainage care in osteomyelitis associated with mandibular fractures.

• 대한소아치과학회지
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• 제30권1호
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• pp.80-91
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• 2003

치아우식증의 원인균인 S. mutans GS5와 S. sobrinus 6715에 대한 polyP의 효과를 관찰하여 보다 안전하고 효과적인 치아우식증 예방을 위한 임상적용의 가능성을 고찰하고자 첫째, 다양한 사슬길이의 polyP를 첨가한 후 흡광도를 측정하여 MIC를 결정하고, 둘째, 실험균주를 흡광도 $0.3{\sim}0.5$까지 증식시킨 후 MIC 농도의 polyP를 첨가하여 흡광도의 변화를 측정함으로써 균주증식 후 성장 억제효과를 관찰하였으며, 셋째, 생균수 측정으로 polyP의 항균효과를 평가하였고, 넷째, 핵산유리의 정도로 polyP의 킬레이션 작용여부를 관찰하였으며, 다섯째, polyP의 비수용성 글루칸 합성능력을 관찰하였으며, 여섯째, 투과전자현미경으로 세포막과 세포질 내의 구조적 변화를 관찰하였다. 이상의 연구를 통하여, polyP의 살균작용이 S. mutans와 S. sobrinus에 대한 성장을 억제시키는 효과가 있는 것으로 가늠된다. 이와 같은 성장 억제효과는 polyP의 킬레이션에 의한 것이라기보다는 균주 세포의 구조적, 형태적 변화가 주된 요인이었던 것으로 판단된다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권8호
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• pp.4031-4036
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• 2012

Background: The negative signaling provided by interactions of the co-inhibitory molecule, programmed death-1 (PD-1), and its ligands, B7-H1 (PD-L1) and B7-DC (PD-L2), is a critical mechanism contributing to tumor evasion; blockade of this pathway has been proven to enhance cytotoxic activity and mediate antitumor therapy. Here we evaluated the anti-tumor efficacy of AAV-mediated delivery of the extracellular domain of murine PD-1 (sPD-1) to a tumor site. Material and Methods: An rAAV vector was constructed in which the expression of sPD-1, a known negative regulator of TCR signals, is driven by human cytomegalovirus immediate early promoter (CMV-P), using a triple plasmid transfection system. Tumor-bearing mice were then treated with the AAV/sPD1 construct and expression of sPD-1 in tumor tissues was determined by semi quantitative RT-PCR, and tumor weights and cytotoxic activity of splenocytes were measured. Results: Analysis of tumor homogenates revealed sPD-1 mRNA to be significantly overexpressed in rAAV/sPD-1 treated mice as compared with control levels. Its use for local gene therapy at the inoculation site of H22 hepatoma cells could inhibit tumor growth, also enhancing lysis of tumor cells by lymphocytes stimulated specifically with an antigen. In addition, PD-1 was also found expressed on the surfaces of activated CD8+ T cells. Conclusion: This study confirmed that expression of the soluble extracellular domain of PD-1 molecule could reduce tumor microenvironment inhibitory effects on T cells and enhance cytotoxicity. This suggests that it might be a potential target for development of therapies to augment T-cell responses in patients with malignancies.

• 농업과학연구
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• 제14권2호
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• pp.286-294
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• 1987

오골계 난백 lysozyme의 용균특성을 조사하고, 이를 식품보존료로 이용하기 위한 자료를 얻고자 오골계 난백으로부터 추출 분리한 lysozyme의 각종 미생물에 대한 용균성을 조사하였다. 그 결과, 공시균중 Gram 양성균인 Staphylococcus aureus phage type 29와 Bacillus subtilis ATCC 6633에 대하여는 용균성이 인정되었으나, Gram 음성균인 E. coli를 비롯한 여타의 균종과 Staphylococcus aureus phage type 57은 lysozyme에 대한 용균 감수성이 인정되지 않았다. 분리된 lysozyme은 S. aureus phage type 29를 공시균주로 하여 이를 육즙배지에 접종하고, $37^{\circ}C$에서 24시간 정치배양하여 세포를 회수한 후 540nm에서 흡광도가 0.6이 되도록 0.05M 초산완충액(pH 4.5)에 현탁시켜, 여기에 0.05%의 lysozyme을 가하여 $30^{\circ}C$, 30분간의 조건으로 반응시킬 때 용균성이 가장 좋았다. 또한 0.05%의 lysozyme은 반응액 중에 glycine(1%)을 첨가하므로서 공시균에 대한 용균효과가 양자의 상승작용에 의하여 그 단용시보다 현저히 증대(<50%) 되었으나, 기타 다른 첨가제와 금속이온 및 lysozyme과의 혼용효과는 인정되지 않았다.

• 대한수의사회지
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• 제25권10호
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• pp.595-606
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• 1989

항원제시세포(APC)와 보조T세포 간의 협력작용에 의하여 활성화된 작동세포(NK세포, CTL, K세포, 대식세포, 과립구 등)의 종양세포, 이식장기 및 세포내기생세균에 감염된 각종 세포에 대한 세포독성작용은 생체방어를 위한 중요한 세포성면역기전이다. 지난 몇 년간 세포성면역기전에 관한 많은 연구에도 불구하고 T림파구매개성 세포독성작용의 면역생물학적기전은 확실히 밝혀있지 않다. 지금까지 알려진 중요한 연구내용을 요약하면 다음과 같다. 1. 세포독성작용을 나타내는 작동세포로는 NK세포, CTL, K세포, 대식세포/단핵구 및 과립구가 있다. 2. T세포의 세포표면항원분자군(CD)으로는 $CD_{2},\;CD_{3},\;CD_{4}[Ly_{3}T_{4}],\;CD_{5}[=Ly_{1}],\;CD_{7},\;CD_{8}[Ly_{2,3}]$가 있으며 $CD_{4}$는 보조Ttpvhdml 특이마커이고 $CD_{8}$는 세포독성 T세포 및 억압T세포의 특이마커이다. 주요 T세포수용체(TCR)는 $CD_{4}$ 또는 $CD_{8}$ 분자와 가까이 연합된 이향체($TCR-{\alpha}{\beta}/TCR-{\gamma}{\delta}$이며 보조 T세포 $CD_{4}$(마우스 $L_{3}T_{4}$)는 수용체와 연합되어 있는반면 억압 T세포 $CD_{8}(Ly+_{2,3})$는 항원수용체와 연합되어 있다. 3. T세포는 Ti-$CD_{3}$(항원/MHC) 복합체를 통한 '항원가교'에 의한 자극(항원인식)과 $CD_{2}$를 통한 비특이경로에 의하여 활성화(분화증식)된다. 비특이경로를 통한 활성경로에서 T세포($CD_{4}$ 및 $CD_{8}$)가 활성화되기 위하여는 보조T세포가 생산하는 IL-2을 요구하며 IL-2의 자극으로 분화증식된 $CD_{8}$는 세포독성능을 나타내지만 $CD_{4}^{+}$는 여전히 세포독성능을 나타내지 못한다. 4. 보조T세포는 class II MHC분자와 연합된 항원을 식별하는 반면 세포독성T세포는 class I MHC 분자와 연합된 항원을 식별한다. 5. 림파구 매개성 세포독성은 접촉(conjugati-on), 탈분극(depolarization), 용해계획(progra-mming), 용해(lysis) 및 재순환(recycling)의 단계를 거쳐 진행된다. 6. 표적세포살해매체로는 perforin / PFP / cy-tolysin, lymphopores, lymphotoxins, protone, cytolytic enzymes가 알려졌으며 세포독성작용은 이들 이외에도 여러 가지 매체를 통한 복합작용으로 추정된다. 7. CTL 매개성 표적세포의 주요 대사변화는 actomyocin ATPase의 증가, phosphocreatine과 ATPase의 소모, ATP 의존성 $Na^{+}/K^{+}$ 펌프작용의 중지, ATP 의존성 $Ca^{2+}$ 유출감소 및 세포내 축적이 관찰된다. 8. $Ca^{2+}$의 축적으로 세포막 교질 침투손상을 주어 수분의 유입을 증가시킴으로써 수포형성, 핵붕괴, 사립체팽화 및 정상세포 구조상실(Zeiosis)이 있다. 결론적으로 CTL 매개성 세포독성작용은 PFP, LT, TNF, 유사 TNF / LT 및 기타 매체를 통한 복합작용이며 세포살해기전은 지속적 대사소모와 정형적 세포구조(핵 및 세포질)의 파괴에 의한 것이다.