제주도 연근해 상어육의 중금속 농도 및 식품안전성

학위논문(석사)--서울대학교 대학원 :수의과대학 수의학과,2019. 8. 박세창.상어고기는 아시아를 필두로 하여 전 세계 다양한 국가에서 식용자원으로 이용되어 왔다. 그러나, 이들은 해양생태계의 최상위층에 존재하는 포식자이기 때문에 중금속 생물축적에 취약하다. 인간의 소비를 위해 연간 1억마리 이상의 상어가 포획되고 있는 만큼, 상어고기의 안전성 판단은 중요한 과제이다. 본 연구에서는 한국 제주도 연근해에 서식하는 6종 (이주성 상어 3종: Carcharhinus brachyurus, Carcharhinus obscurus, Isurus oxyrinchus; 정착성 상어 3종: Triakis scyllium, Mustelus manazo, Cephaloscyllium umbratile) 상어 근육조직의 중금속 농도를 분석하였다. 총 11종의 중금속 (Cr, Fe, Cu, Zn, As, Se, Cd, Sn, Sb, Pb, Hg) 및 MeHg의 농도를 ICP-MS, DMA, GC 및 CVAFS를 이용하여 확인하고 국제식품규격위원회를 포함하여 다양한 기관에서 제시하고 있는 최대규제치 값과 비교하였다. 그 결과 As를 제외한 모든 중금속의 평균 농도는 규제치 이하인 것으로 나타났다. 그 중 Hg와 MeHg는 체장, 체중 및 나이와 유의미한 양의 상관관계를 보였으며, C. brachyurus 종의 경우 체장 130cm, 체중 25kg 이상인 개체에서는 높은 확률로 Hg 농도가 규제치를 초과할 것으로 계산되었다. 본 연구 결과 상어고기를 소비할 경우 소비자들은 높은 농도의 As에 노출될 수 있으며, 예측된 크기보다 큰 C. brachyurus유래 고기를 소비하는 경우 또한 높은 농도의 Hg에 노출될 수 있음이 확인되었다. 추가적인 연구를 통해 다양한 상어고기의 소비 시 적용할 세밀한 규제 및 가이드라인을 설정해야 할 필요성이 제시된다.Shark meat is consumed as a food source worldwide, especially in Asian countries. However, since sharks are apex predators in the ocean food chain, they are prone to bioaccumulation of heavy metals. More than 100 million sharks are caught annually for human consumption, and the safety of shark meat cannot be overemphasized. Here, we examined heavy metal concentration in the muscle tissue of 6 shark species including 3 migratory species (Carcharhinus brachyurus, Carcharhinus obscurus, and Isurus oxyrinchus) and 3 local species (Triakis scyllium, Mustelus manazo, and Cephaloscyllium umbratile) from fish markets in Jeju Island, Republic of Korea. The concentrations of 11 heavy metals (Cr, Fe, Cu, Zn, As, Se, Cd, Sn, Sb, Pb, and Hg) and MeHg were analyzed. The result showed that the average concentrations of all metals, except for that of As, were below the regulatory maximum limits of many organizations, including the Codex standard. Hg and MeHg were significantly correlated with body length, body weight, and age, and the concentration of Hg was expected to exceed the limit in C. brachyurus with a body length or weight of over 130 cm or 25 kg, respectively. Our results indicate that shark meat can expose consumers to a high level of As and that copper sharks bigger than the predicted size should be avoided for excessive Hg. Considering these findings, a detailed guideline on consumption of meat of different shark species should be suggested based on further investigation.Abstract∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ i Contents∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ iii Abbreviations∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ iv Introduction∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 1 Material & Methods∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 7 Results∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 16 Discussion∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 26 References∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 47 Summary∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 55 Abstract in Korean ∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 56 List of articles∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 58 List of conferences∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 62 Acknowledgements∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙∙ 66Maste

상어에서의 기초 보조생식기술 연구

학위논문(박사) -- 서울대학교대학원 : 수의과대학 수의학과, 2022.2. 박세창.상어는 판새아강 상어상목에 속하는 어종들의 총칭으로, 초기 척추동물 중 현재까지 성공적으로 생존해 있는 대표적인 동물군이다. 척추동물 형성 초기에 나타나 진화의 역사를 고스란히 담고 있는 만큼, 상어는 면역체계, 번식 전략, 암 저항성 등 다양한 측면에서 중요한 진화생물학적 연구의 대상이다. 뿐만 아니라 상어는 생태학적으로도 중요한 역할을 하는데, 해양 생태계 먹이사슬의 최상위 포식자로서 해당 생태계가 균형 있고 안정적으로 유지될 수 있도록 기여하고 있기 때문이다. 상어 개체 수에 이상이 생기면 먹이사슬 전체가 무너질 수 있다는 점은 이미 여러 선행연구를 통해 밝혀진 바, 그 중요성이 높기 때문에 핵심종이라고도 불린다. 문제는 이들이 현재 심각한 멸종위기에 처해있다는 점이다. IUCN Red List에 따르면, 연골어류의 37%가량이 취약 (VU), 절멸 위기 (EN), 절멸 위급 (CR)군으로 분류되고 있어 심각한 멸종위기 상태를 마주하고 있다. 이러한 양상은 이미 1970년대부터 확인되어 왔으며, 그 가장 큰 요인으로는 샥스핀 산업을 필두로 하는 어업이 지적되어 왔다. 이에 전세계적으로 상어 어획량을 줄이고자 하는 환경운동이 수행되어 왔으나, 상어 특유의 느린 번식속도에 기인하여 이들의 멸종위기 상태가 쉽게 개선되지는 않고 있는 실정이다. 심각한 멸종위기 상태의 동물 종 보전을 위해서는 사람의 인위적인 개입이 필수불가결하다. 이미 재두루미, 몽고말, 코끼리, 북부흰코뿔소 등 다양한 멸종위기 동물종에서 종 구제 및 보호를 위하여 보조생식기술이 개발 적용되고 있다. 그러나 상어의 경우 동물 자체에 대한 접근이 쉽지 않아 사실상 연구 되어있는 바가 거의 없다. 이에 본 논문에서는 상어 종 보전에 기여하고자 일련의 연구들을 수행하였다. 우선적으로 기초적인 영상의학 아틀라스를 확립하였으며, 이후 보조생식기술 개발을 위하여 상어 정액 동결보존 프로토콜 및 호르몬 유도 배란 프로토콜, 호르몬 유도 정액 샘플링 프로토콜을 개발하였다. 1. 영상의학적 분석 기반 확립은 모든 수의학적 접근 방식의 기본이 되는 만큼, 본 연구는 최초로 컴퓨터 단층촬영 (CT) 및 자기공명영상진단 (MRI) 방식을 이용하여 상어에서의 세밀한 영상의학 아틀라스를 확립하였다. 체장 1 m 안팎의 어린 까치상어 (Triakis scyllium) 세 마리의 전신 CT 및 MRI 스캔을 수행했으며, 각 개체들을 냉동 후 transverse, sagittal, dorsal 단면으로 잘라 실제 단면 모습과 CT, MRI상의 단면을 비교분석 하였다. 다양한 장기 및 조직들을 세밀하게 구분하여 아틀라스를 확립하였다. 다만 미성숙 개체들을 대상으로 연구를 수행하였던 만큼 생식계통에 대한 확인은 불가하였다. 2. 까치상어 (Triakis scyllium) 수컷 다섯 마리를 대상으로 실험을 수행하였다. Ovaprim®을 0.2 mL/kg 투여 후 1시간 뒤, 상어의 복부를 부드럽게 마사지하여 urogenital papilla를 통해 나오는 정액의 secondary cloudy portion을 샘플링 하여 실험에 이용하였다. 정지상태 정자의 motility를 최대로 만들 수 있는 활성화 용액 조성을 확립하였으며 이를 SSAE-1로 명명하였다. 까치상어 정자에 최적화된 동결보존 프로토콜을 확립하기 위하여 총 8종의 extender solution, 3종류의 extension ratio, 15종의 cryoprotectants, 4종의 equilibration periods, 3종의 cooling rates, 3종의 thawing temperature를 테스트 하였다. 결과적으로 정립된 Kim’s protocol은 다음과 같다: extender, filtered seawater; extension ratio, 1:3; cryoprotectant, egg yolk 10% + ethylene glycol 10%; equilibration period, 10 min; cooling rate, 3 cm, 3 min; thawing temperature, 30℃, 10 s. 프로토콜을 이용했을 때 최종적으로 확인된 정자의 해동 후 운동능은 2.03%로 확인되었다. 3. 난태생 상어인 까치상어 (Triakis scyllium)와 태반성 태생 상어인 화이트팁 상어 (Triaenodon obesus)에서 실험을 수행하여 다양한 번식전략을 지닌 상어 종 전반에 걸쳐 연어 유래 gonadotropin-releasing hormone (GnRH, Ovaprim®) 적용 가능성을 확인하였다. 실험에 앞서 두 상어종에서 암컷과 수컷의 정상 혈중 성호르몬 농도를 각각 확인하여 추후 실험 분석의 base line으로 이용하였다. 두 종 모두에서 Ovaprim®이 혈중 에스트로겐, 프로게스테론, 테스토스테론의 농도 변화를 성공적으로 유도하였으며, 암컷에서는 난포 성숙 및 배란을, 수컷에서는 정액 사출을 유도함을 확인하였다. 추후 인공수정에 적용할 수 있도록 Ovaprim®의 농도 및 투여 주기를 최적화한 결과는 다음과 같았다: 까치상어 수컷: 0.2 mL/kg 투여 1시간 뒤 정액 샘플링; 까치상어 암컷: 0.2 mL/kg 투여 24시간 뒤 0.5 mL/kg 2차 투여; 화이트팁 상어 수컷: 0.2 mL/kg 투여 직후 정액 샘플링; 화이트팁 상어 암컷: 0.2 mL/kg 투여 24시간 뒤 0.2 mL/kg 또는 0.3 mL/kg 2차 투여.Shark is a generic term for fish species belonging to the Class Chondrichthyes Superorder Selachimorpha and is a representative animal group that has successfully survived to date among early vertebrates. As they appeared in the early stages of vertebrate formation and thus contain the history of evolution, sharks are important subject of evolutionary biological studies in various aspects such as immunology, reproductive biology, and cancer biology. In addition, sharks play an important role in ecological point of view because they are the apex predators of the marine food chain and are contributing to maintain a balanced and stable ecosystem. It has already been revealed through several previous studies that if the number of sharks become seriously reduced, the entire food chain of the relevant area can collapse. Sharks are thus also called as keystone species because of their high importance. The problem is that sharks are currently in critical danger of extinction. According to the IUCN Red List, about 37% of chondrocytes are classified into “vulnerable (VU),” “endangered (EN),” and “critically endangered (CR)” groups, facing serious extinction. Decline of population had been perceived since the 1970s, and the biggest contributing factor has been pointed out as fishing, led by the shark's fin industry. Accordingly, conservational efforts have been carried out worldwide to reduce shark fishing, but their endangered status has not been easily improved due to their uniquely slow breeding rate. Artificial intervention by human is indispensable for the conservation of critically endangered animal species. Assisted reproductive technology has already been developed and applied to conservational works for various endangered species such as white cranes, Przewalski’s horses, elephants, and northern white rhinos. However, in the case of sharks, access to the animals is not easy, so few studies have been conducted thus far. Therefore, in this paper, a series of studies on assisted reproductive technology were conducted to contribute to the shark conservation. First, basic imaging atlas was established, and then shark semen cryopreservation protocol, hormone induced ovulation protocol, and hormone induced semen sampling protocol were developed. 1. As imaging analysis techniques are the basis of veterinary approaches, this study established detailed imaging atlas in sharks using computed tomography (CT) and magnetic resonance imaging (MRI) methods for the first time. Whole-body CT and MRI scans were performed with three young banded houndsharks (Triakis scyllium) of around 1 m in total body length, and each individual was cryosectioned into transverse, sagittal, and dorsal planes to compare and analyze with the images from CT and MRI scans. Atlas was established by classifying various organs and tissues in detail. However, it was impossible to confirm the reproductive system as the study was conducted on immature individuals. 2. Experiments were conducted on five male banded houndsharks (Triakis scyllium). One hour after 0.2 mL/kg of Ovaprim® administration, the shark's abdomen was gently massaged and the secondary cloudy portion of semen was sampled through urogenital papilla. The composition of an activating extender capable of maximizing the motility of stationary spermatozoa was established, which was designated as SSAE-1. To establish a cryopreservation protocol optimized for banded houndshark semen, a total of 8 extender solutions, 3 extension ratios, 15 cryoprotectants, 4 equilibration periods, 3 cooling rates, and 3 thawing temperatures were tested. The optimized protocol (Kim’s protocol) was as follows: extender, filtered seawater; extension ratio, 1:3; cryoprotectant, egg yolk 10% + ethylene glycol 10%; equilibration period, 10 min; cooling rate, 3 cm, 3 min; thawing temperature, 30℃, 10 s. The resulting post-thaw spermatozoa motility was 2.03%. 3. Experiments were conducted on the ovoviviparous shark banded houndshark (Triakis scyllium) and the placental shark (Triaenodon obesus) to confirm the applicability of salmon-derived gonadotropin-releasing hormone analogue (sGnRHa, Ovaprim®) throughout shark species with various breeding strategies. Prior to the experiment, normal blood sex hormone concentrations of females and males were identified in each of the species, and used as the base line for further experimental analysis. In both species, it was confirmed that Ovaprim® successfully induced changes in concentration of estradiol, progesterone, and testosterone in the blood, follicular maturation and ovulation in females, and semen release in males. The optimized injection protocols of Ovaprim® for future application to artificial insemination were as follows: male banded houndshark: 0.2 mL/kg administration and semen sampling 1 hour after administration; female banded houndshark: 0.2 mL/kg first administration and 0.5 mL/kg second administration with 24 hours of gap time; male whitetip reef shark: 0.2 mL/kg administration and semen sampling right after administration; female whitetip reef shark: 0.2 mL/kg first administration and 0.2 mL/kg or 0.3 mL/kg second administration with 24 hours of gap time.ABSTRACT CONTENTS LIST OF FIGURES LIST OF TABLES ABBREVIATIONS CHAPTER I 1 Abstract 2 1. Introduction 3 2. Materials and Methods 5 3. Results 8 4. Discussion 9 5. Conclusion 13 References 39 CHAPTER II 47 Abstract 48 1. Introduction 49 2. Materials and Methods 52 3. Results 59 4. Discussion 63 5. Conclusion 73 References 88 CHAPTER Ⅲ 97 Abstract 98 1. Introduction 99 2. Materials and Methods 103 3. Results 112 4. Discussion 118 References 141 GENERAL CONCLUSION 151 국문 초록 153 PUBLISHED ARTICLES 158 CONFERENCE PROCEEDINGS 166 감사의 글 168박

Protective effects of renal fibrosis by renal-specific chitobionic acid- conjugated polysorbitol gene transporter carrying hepatocyte growth factor

학위논문 (박사)-- 서울대학교 대학원 의과대학 협동과정 종양생물학전공, 2017. 8. 강경선.Chronic kidney disease (CKD) refers to kidney disease that results in structural or functional damage to the kidneys due to a variety of causes, resulting in irreversible loss of nephron and renal function. The incidence and mortality rate of CKD is increasing worldwide, due to the absence of treatment methods and the existence of various causes of disease. Renal fibrosis is a chronic and progressive kidney disorder that is associated with many kidney diseases. Recovery from acute kidney injury involves the regeneration of the tissue, in which the damaged tissue is replaced by new cells of the same lineage. Abnormal or incomplete recovery lead to renal fibrosis, which is a cause of chronic, progressive renal failure. Considerable research has focused on the development of efficient and specific methods to treat renal diseases. Gene therapy using gene carriers has the advantages of high transfer efficiency, low toxicity, and application to various diseases. In addition, non-viral gene carriers also have potential for gene therapy, due to their ease of modification and biocompatibility. This study was demonstrated that the protective effects of kidney – specific chitonic acid-conjugated polysorbitol gene transporter (CBA-PSGT) carrying the gene for hepatocyte growth factor (HGF) as an anti fibrotic factor, against renal fibrosis progression. First, CBA, a kidney specific ligand, was synthesized, and conjugated it to a PSGT carrier. CBA can effectively target vimentin to the kidney. Vimentin is an intermediate filament protein that is found in vascular endothelial cells, vascular smooth muscle cells and fibroblasts. Therefore, vimentin, which is expressed on the surface of kidney cells or kidney tissues, can be used as a target for gene delivery. Moreover, vimentin is expressed in the renal interstitial region, as well as in normal and abnormal renal tissues. In particular, vimentin can be considered as a marker of renal tubular disorder, owing to its high expression in those disorders, including renal atrophy, inflammation, and fibrosis. The kidney-specific gene transporter CBA-PSGT as a gene carrier was synthesized and evaluated. CBA-PSGT was confirmed a successful gene transporter with high stability, due to DNA complexation, minimal toxicity, and high gene expression efficiency with the gene. In addition, CBA-PSGT was effectively protected from digestion by DNase. Second, unilateral ureteral obstruction (UUO) animal models were constructed for the evaluation of the protective effects of CBA-PSGT/HGF against renal fibrosis. In the UUO model, the expression of HGF was increased by treatment with CBA-PSGT/HGF. The UUO model causes renal tissue damage, mainly resulting in atrophy of the interstitial tissue of the tubule and fibrosis. In addition, infiltration of inflammatory cells into the tubulointerstitial tissue causes inflammatory cytokine release in the UUO model. The CBA-PSGT/HGF delivery group was detected less damage to the kidney tissue by histological analysis. BUN and creatinine levels were used as kidney damage markers, and the expression levels of inflammatory cytokines, interleukin-6 (IL-6) and IL-1β were deceased in the group that treated of CBA-PSGT/HGF. Renal fibrosis lead to the formation and accumulation of collagen in the renal tissues. Therefore, collagen formation and accumulation were reduced by gene therapy by histological analysis and assessment of the levels of collagen. In addition, the expression level of collagen-related proteins, such as ICAM-1, TIMP-1 and α-SMA, which highly expressed in the setting of renal injury, as well as metalloproteinase (MMPs), which are enzymes that inhibits the formation of extraellular matrix (ECM), were decreased in the CBA-PSGT/HGF complex group. In summary, a CBA-conjugated gene transporter was successfully synthesized as kidney specific ligand, which targets the vimentin that is highly expressed in the setting of renal disease and damage. It is characterized that the kidney-specific gene transporter. CBA-PSGT was stable due to DNA complexation and it was protected from degradation by DNasethe construct also had low toxicity and high delivery efficiency. In addition, The study was demonstrated that CBA-PSGT carrying HGF, an anti-fibrotic factor, protected against the progression of renal fibrosis in a UUO model. CBA-PSGT should be further investigated, to explore its potential to alleviate UUO and kidney-related disease using high affinity kidney targeting.Protective effects of renal fibrosis by renal-specific chitobionic acid- conjugated polysorbitol gene transporter carrying hepatocyte growth factor 1 INTRODUCTION 2 1. Gene delivery system 2 1.1. Viral vector gene delivery system 3 1.2. Non-viral vector gene delivery system 6 1.3. Chitosan conjugated gene delivery system 11 2. Kidney fibrosis 13 2.1. Kidney injury animal model 13 2.2. Mechanism of kidney fibrosis 16 2.3. Gene delivery system in kidney disease 20 3. Experimental purpose 23 MATERIALS AND METHODS 24 RESULTS 35 DISCUSSION 74 REFERENCES 81 LIST OF ABBREVIATION 108 ABSTRACT OF KOREAN 110Docto

Next-generation sequencing of BRCA1/2 in breast cancer patients: potential effects on clinical decision-making using rapid, high-accuracy genetic results

PURPOSE: We evaluated the clinical role of rapid next-generation sequencing (NGS) for identifying BRCA1/2 mutations compared to traditional Sanger sequencing. METHODS: Twenty-four paired samples from 12 patients were analyzed in this prospective study to compare the performance of NGS to the Sanger method. Both NGS and Sanger sequencing were performed in 2 different laboratories using blood samples from patients with breast cancer. We then analyzed the accuracy of NGS in terms of variant calling and determining concordance rates of BRCA1/2 mutation detection. RESULTS: The overall concordance rate of BRCA1/2 mutation identification was 100%. Variants of unknown significance (VUS) were reported in two cases of BRCA1 and 3 cases of BRCA2 after Sanger sequencing, whereas NGS reported only 1 case of BRCA1 VUS, likely due to differences in reference databases used for mutation identification. The median turnaround time of Sanger sequencing was 22 days (range, 14-26 days), while the median time of NGS was only 6 days (range, 3-21 days). CONCLUSION: NGS yielded comparably accurate results to Sanger sequencing and in a much shorter time with respect to BRCA1/2 mutation identification. The shorter turnaround time and higher accuracy of NGS may help clinicians make more timely and informed decisions regarding surgery or neoadjuvant chemotherapy in patients with breast cancer.ope

Delayed postpartum regression of theca lutein cysts with maternal virilization: A case report

Theca lutein cysts are rare, benign lesions responsible for gross cystic enlargement of both ovaries during pregnancy. This condition is also termed hyperreactio luteinalis. Elevated human chorionic gonadotropin (hCG) levels or states of hCG hypersensitivity seem to promote these changes, which in up to 30% of patients produce clinical signs of hyperandrogenism. Given the self-limiting course of theca lutein cysts, which are subject to spontaneous postpartum resolution, conservative treatment is the mainstay of patient management. Described herein is a rare case of theca lutein cysts with maternal virilization that failed to regress by 9 months after childbirth. Surgical intervention was eventually undertaken, necessitated by adnexal torsion.ope

유방보존술을 시행한 유방암환자에서 삼중음성유방암의 임상적 영향

의과대학/석사Background and Objectives : The optimum local surgical strategy regarding Breast-Conserving Therapy (BCT) for Triple-Negative Breast Cancer (TNBC) is controversial. We evaluated the clinical outcomes of BCT in women with TNBC compared to those without TNBC, using a large, single center cohort. Methods : We performed a retrospective analysis of 1533 women (TNBC n=321; non-TNBC n=1212) who underwent BCT for primary breast cancer between 2000 and 2010. Clinicopathological characteristics, locoregional recurrence-free survival (LRFS), and overall survival (OS) were analyzed. Results : Tumors from the TNBC group had higher T stage (T2 37.4% vs. 21.0%, p<0.001), lower N stage (N0 86.9% vs. 75.5%, p<0.001), and higher histologic grade (Grade III 66.8% vs. 15.4%, p<0.001) versus the non-TNBC group. There were no differences in 5-year LRFS rates between the TNBC and non-TNBC groups (98.7% vs. 97.8%, p=0.63). The non-TNBC group showed a slightly better 5-year OS than the TNBC group, but the difference was not significant (96.2% vs. 97.3%, p=0.72). In multivariate analyses, TNBC was not associated with poor clinical outcomes in terms of LRFS and OS (HR for LRFS=0.37, 95% CI=0.10-1.31, HR for OS=1.03, 95% CI=0.31-3.39). Conclusions : TNBC patients who underwent BCT showed relatively low locoregional recurrence. BCT is an acceptable surgical approach in selected patients with TNBC.ope

The clinical implication of the number of lymph nodes harvested during sentinel lymph node biopsy and its effects on survival outcome in patients with node-negative breast cancer

BACKGROUND: The optimal number of sentinel lymph nodes (SLN) that need to be harvested to achieve favorable survival outcome during a SLN biopsy (SLNB) has not yet been established. METHODS: Six hundred and thirteen patients with clinically node-negative breast cancer who underwent SLNB were reviewed. Survival outcomes according to the number of total harvested lymph nodes (THLNs), defined as the sum of enumerated SLNs and non-SLNs were analyzed. RESULTS: Patients with only 1 THLN showed lower recurrence-free survival (RFS) as compared to those with ≥2 THLNs (p = 0.049). In multivariate analysis, only 1 THLN was associated with poor RFS (HR = 2.711; p = 0.029). CONCLUSIONS: Removing at least 2 lymph nodes during SLNB may be acceptable. Harvesting only 1 lymph node should be undertaken cautiously because of false negative results and increasing the subsequent recurrence rate.restrictio

Serum 25-hydroxyvitamin D deficiency and increased risk of breast cancer among Korean women: a case-control study

Despite the emerging literature supporting the beneficial role of vitamin D on various health outcomes including carcinogenesis, current evidence on the association between vitamin D and breast cancer is still largely inconsistent. Furthermore, this relationship is particularly under explored among Asian population. We conducted a large case-control study with Korean women. We obtained and compared serum 25-hydroxyvitamin D (25(OH)D) between breast cancer patients (N = 3634) and general population (N = 17,133). Moreover, we further examined the association between serum 25(OH)D and breast cancer risk stratified by menopausal status and hormone receptor (HR) status of the tumor. Adjusted odds ratio (OR) for breast cancer comparing women with deficient level of serum 25(OH)D to women with sufficient level of serum 25(OH)D was 1.27 [95 % confidence interval (CI) 1.15-1.39]. This association did not significantly vary by menopausal status [pre-menopause: 1.26 (95 % CI 1.09-1.45) vs. post-menopause: 1.25 (95 % CI 1.10-1.41)]. When stratified by HR status, the inverse association remained significant in both positive and negative statuses. However, this association was more pronounced in HR-negative breast cancer, particularly with triple-negative breast cancer patients (1.45, 95 % CI 1.15-1.82). Given the growing burden of breast cancer in Asia and dearth of studies examining the association between vitamin D and breast cancer risk in Asian women thus far, this study provides a meaningful evidence for potential preventive effect of vitamin D on breast cancer for this particular population.ope