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Accuracy of Frozen Section Analysis of Sentinel Lymph Nodes for the Detection of Asian Breast Cancer Micrometastasis - Experience from Pakistan

  • Hashmi, Atif Ali;Faridi, Naveen;Khurshid, Amna;Naqvi, Hanna;Malik, Babar;Malik, Faisal Riaz;Fida, Zubaida;Mujtuba, Shafaq
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2657-2662
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    • 2013
  • Background: Intraoperative sentinel lymph node biopsy has now become the standard of care for patients with clinically node negative breast cancer for diagnosis and also in order to determine the need for immediate axillary clearance. Several large scale studies confirmed the diagnostic reliability of this method. However, micrometastases are frequently missed on frozen sections. Recent studies showed that both disease free interval and overall survival are significantly affected by the presence of micrometastatic disease. The aim of this study was to determine the sensitivity and specificity of intraoperative frozen section analysis of sentinel lymph nodes (SLNs) for the detection of breast cancer micrometastasis and to evaluate the status of non-sentinel lymph nodes (non-SLNs) in those patients subjected to further axillary sampling. Materials and Methods: We performed a retrospective study on 154 patients who underwent SLN biopsy from January 2008 till October 2011. The SLNs were sectioned at 2 mm intervals and submitted entirely for frozen sections. Three levels of each section submitted are examined and the results were compared with further levels on paraffin sections. Results: Overall 40% of patients (62/154) were found to be SLN positive on final (paraffin section) histology, out of which 44 demonstrated macrometastases (>2mm) and 18 micrometastases (<2mm). The overall sensitivity and specificity of frozen section analysis of SLN for the detection of macrometastasis was found to be 100% while those for micrometastasis were 33.3% and 100%, respectively. Moreover 20% of patients who had micrometastases in SLN had positive non-SLNs on final histology. Conclusions: Frozen section analysis of SLNs lacks sufficient accuracy to rule out micrometastasis by current protocols. Therefore these need to be revised in order to pick up micrometastasis which appears to have clinical significance. We suggest that this can be achieved by examining more step sections of blocks.

A Study for Predicting Adfreeze Bond Strength from Shear Strength of Frozen Soil (동결토 전단강도를 활용한 동착강도 산정에 관한 연구)

  • Choi, Chang-Ho;Ko, Sung-Gyu
    • Journal of the Korean Geotechnical Society
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    • v.27 no.10
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    • pp.13-23
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    • 2011
  • Bearing capacity of pile foundations in cold region is dominated by adfreeze bond strength between surrounding soil and pile perimeter. It denotes that adfreeze bond strength is the most important design parameter for foundations in cold region. Adfreeze bond strength is affected by various factors like 'soil type', 'frozen temperature', 'normal stress acting on soil/pile interface', 'loading rate', 'roughness of pile surface', etc. Several methods have already been proposed to estimate adfreeze bond strength during past 50 years. However, most methods have not considered the effect of normal stress for adfreeze bond strength. In this study, both freezing temperature and normal stress have been controlled as primary factors affecting adfreeze bond strength. A direct shear box was used to measure adfreeze bond strength between sand and aluminum under different temperature conditions. Based on the test results, the relation between shear strength of frozen sand and adfreeze bond strength have been investigated. The test results showed that both of shear strength and adfreeze bond strength tend to increase with decreasing frozen temperature or increasing confining pressure. The ratio of shear strength and adfreeze bond strength, expressed as $r_s$, decreased initially frozen section but increased at much lower frozen temperature and there were uniform intervals under the different normal stress conditions. A method for predicting adfreeze bond strength using $r_s$ has finally been proposed in this study.

Physicochemical Changes of Pork During its Frozen Storage (동결(凍結) 돈육(豚肉) 저장(貯藏) 중(中)의 변화(變化))

  • Park, Seok-Won;Kang, Tong-Sam;Mon, Byong-Yong;Suh, Kee-Bong;Yang, Ruyng
    • Korean Journal of Food Science and Technology
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    • v.12 no.1
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    • pp.34-40
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    • 1980
  • In order to establish the frozen storage method of pork and contribute to the stabilization of pork price physicochemical changes and sensory evaluation of pork, stored at $-20^{\circ}C$ up to the period of 12 months, were analyzed every three months. The drip loss of frozen meat cuts was below 1% regardless of storage months. In the course of storage, pH of frozen half carcass rose a little, while that of meat cuts remained almost the same. WHC(water holding capacity) of frozen half carcass and meat cuts was in the range of $50{\sim}60\;and\;55{\sim}62%,$ respectively and VBN (volatile basic nitrogen) was about $11{\sim}18mg%,$ all of which did considerably change during the storage. TBA(thiobarbituric acid) value was not increased up to the 6th month of storage, but represented a considerable increase after the 9th month of storage, Both tenderness and juiciness of frozen pork were decreased after the 12th month of storage but the axxrptability of frozen pork to the consumers turned out fairly good.

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Studies of the Ultrarapid Freezing of In Vitro Fertilized Bovine Embryos I. Studies on the Survival Rates after Rapid Frozen-Thawing of In Vitro Fertilized Bovine Embryos (소 체외수정란의 초급속동결에 관한 연구 II. 소 체외수정란의 초급속동결 융해후의 생존성에 관한 연구)

  • 김상근;이만휘
    • Korean Journal of Animal Reproduction
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    • v.15 no.2
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    • pp.141-147
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    • 1991
  • This stduy was carried out in order to investigate the effects of cryoprotective concentration and equilibration time on survival rate of ultrarapidly frozen in vitro fertilized bovine embryos. In vitro fertilized bovine embryos, following dehydration by cryoprotective agents and sucorese were directly plunged into liquid nitrogen and thawed in 38$^{\circ}C$ water. Survival rate was defined by development rate to the morula and blaqstocyst stage after in vitro culture of by FDA test. The results are summarized as follows : 1. The survival rates of in vitro fertilized bovine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucroese added 2.0M, 2.5M, 3.0M, 3.5M, 4.0M glycerol were 75.0%, 72.0%, 67.6%, 44.8% and 18.3% respectively. 2. The survival rates of in vitro fertilized bovine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0M, 2.5M, 3.0M, 3.5M, 4.0M DMSO were 64.0%, 66.7%, 70.8%, 52.7% and 18.6, respectively. 3. The survival rates of in vitro fertilized bovine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0M, 2.5M, 3.0M, 3.5M, 4.0M propanediol were 68.4%, 64.9%, 63.2%, 62.2% and 34.7%, respectively. 4. The survival rates of in vitro fertilized bovine embryos after ultrarapid frozen-thawing in the freezing medium of 2.50M glycerol added 0.1M, 0.25M, 0.5M, 0.75M, sucrose were 60.5%, 72.2%, 70.1% and 54.9%, respectively. The survival rate of in vitro fertilized embryos after ultrarapid frozen-thawing in the freezing medium of 2.5M glycerol added 0.25M sucrose were higher than concentration of 0.10M, 0.50M and 0.75M sucrose. 5. The equilibration time on the survival rate of in vitro fertilized bovine embryos was attained after short period of time(2.5~5min.) in the freezing medium added 0.25M sucrose and 3.0M DMSO higher than long period time(1~20min.).

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The Effects of the Ash Content in Flour on the Rheological Properties of Frozen Dough (밀가루의 회분 함량이 냉동 생지 반죽의 물성에 미치는 영향)

  • Kim, Seok-Young;Han, Jae-Heung;Song, Young;Lee, Si-Kyung
    • Applied Biological Chemistry
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    • v.46 no.1
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    • pp.39-45
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    • 2003
  • This study was conducted to investigate the effect of ash contents of bread flour on the rheology of frozen dough In making frozen dough by measuring amylograph, flrinograph and extensograph. The quality of frozen-stored dough under freezing condition ($-20^{\circ}C$, 12 weeks) was evaluated by measuring final proof time, moisture content, baking loss, loaf volume and hardness of bread with storage time. In bread flour with high ash content farinogram showed that water absorption, degree of softening increased but valorimeter value decreased. In bread flour with low ash content amylogram showed that gelatinization temperature and maximum viscosity increased and extensogram showed that the area and resistance of the bread flour increased. As the proof time increased the extensibility decreased. Final proof time of frozen dough was shortened at the bread flour with low ash content with storage time. In bread using the flour with high ash content, moisture content, increased but baking loss rate decreased while the hardness of product increased slowly with time. But in bread using the flour with low ash content, the loaf volume of baking increased but the hardness of product decreased. As the frozen storage time was shortened, the product was more stable and better in quality.

Studies on In Vitro Developmental Rate of Activated Bovine Oocytes by Intracytoplasmic Sperm Injection with Frozen-Thawed Epididymal Spermatozoa (정자미세주입술에 의하여 동결 융해 부고환 정자와 수정시킨 활성화처리 난자의 체외발생율에 관한 연구)

  • 김상근;이동수
    • Journal of Embryo Transfer
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    • v.17 no.1
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    • pp.55-59
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    • 2002
  • The objective of this study is to determine the developmental competence of in vitro matured bovine oocytes after intracytoplasmic sperm injection(ICSI) with frozen-thawed epididymal spermatozoa. The ovaries were obtained from slaughtered Korean native cows. Oocytes matured in vitro for 24 hrs were fertilized by ICSI with frozen-thawed epididymal spermatozoa. After ICSI, a group of oocytes was activated with 7% ethanol fur 5 min, and the other group was not activated. The oocytes were cultured in TCM-199 medium containing hormones and 10% FCS for 24~30 hrs in a incubator with 5% $CO_2$ in air at 38.5$^{\circ}C$. The percentage of oocytes reaching M II after 24 hrs and 30 hrs of incubation were significantly higher(p<0.05) after culture with TCM-199 media(80.0% and 88.3%) than M I(8.3% and 6.7%). The rate of cleavaged embryos to blastocyst obtained by ICSI treated activation oocytes was significantly higher(p<0.05) than that of nonactivation oocytes(22/46, 47.8% vs 10/39, 25.6%). The rates of embryos development to blastocyst obtained by ICSI treated sperm of flesh, epididymal and frozen-thawed epididymal were 24/45(53.3%), 15/40(37.5%), 11/43(25.6%), respectively and these values of fresh sperm injection were higher than frozen-thawed epididymal sperm. We also concluded that embryos can be produced with ICSI of in vitro matured oocytes by ICSI using frozen-thawed epididymal semen.

Production of cloned Rabbits Embryos and Offsprings by Nuclear Transplantation using In Vitro Matured Oocytes in Rabbits (토끼의 체외배양 난자를 이용한 핵이식으로 복제수정란 및 복제산자의 생산)

  • 박충생;전병균;이경미;윤희준;이효종;곽대오;최상용
    • Journal of Embryo Transfer
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    • v.10 no.1
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    • pp.65-72
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    • 1995
  • The purposes of this study were to produce cloned rabbit embryos and offsprings by nuclear transplantation(NT) using in vitro matured oocytes as nuclear recipient cytoplasm and to determine the effect of frozen nuclei donor embryos on the production efficiency of cloned embryos. The 8cell embryos were collected from the mated does by flushing oviducts with Dulbecco's phosphate buffered saline containing 10% fetal calf serum(FCS) at 40 hours after hGG injection. A portion of collected embryos were preserved at 4$^{\circ}C$ for 24 hours and a portion of them were frozen by vitrification method. The embryos used for donor nuclei were synchronized in the phase of Gi /S transition. The in vitro matured oocytes were used as recipient cytoplasm following removing the nucleus and the first polar body. The synchronized blastomeres from fresh, cooled or frozen embryos were injected into the enucleated oocytes by micromanipulation and were electrofused by electrical stimulation of three pulses for 60 $\mu$sec at 1.0 W /cm in 0.28 M mannitol solution. The fused oocytes were co-cultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FCS for 120 hours at 39$^{\circ}C$ in a 5% $CO_2$incubator. Following in vitro culture of the NT embryos to blastocyst stage, they were stained with Hoechst 33342 dye for counting the number of blastomeres by fluorescence microscopy. The nuclear transplant embryos developed in vitro to 2- to 4-cell stage were transferred into the oviducts of synchronized recipient does. The results obtained were summarized as follows: 1. The fusion rates of the blastomeres from fresh, cooled and frozen embryos with the in vitro matured and enucleated oocytes were 100, 95.8 and 64, 3%, respectively. 2. Development in vitro to blastocyst was significantly(p<0.05) different between the cloned embryos with the blastomeres from fresh, cooled or frozen embryos as 39.0, 20. 9 and 15.7%, respectively. 3. The mean numbers of cell cycle per day during in vitro culture of cloned embryos blastomeres from fresh, cooled or frozen embryos was 1.31, 1.29 and 1.16, respectively. 4. A total of 77 nuclear transplant embryos were transferred into 6 recipient does, of which two offsprings were produced from a foster mother 31 days after embryo transfer.

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Development of New Vitrification Method for Preimplantation Mouse Embryo

  • Ha, A-Na;Fakruzzaman, Md.;Lee, Kyeong-Lim;Wang, Erdan;Lee, Jae-Ik;Min, Chan-Sik;Kong, Il-Keun
    • Journal of Embryo Transfer
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    • v.28 no.2
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    • pp.141-147
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    • 2013
  • The purpose of this study was attempted to new methods in mammalian embryos vitrification. This method was affected to increase of the embryo vitrification efficiency and it would be applied to the field of embryo transfer to recipient by modified loading method of embryo into 0.25 ml plastic straw. The frozen mouse embryos were carried out warmed from two different cell stages (8-cell and blastocyst, respectively) by attachment of an embryo in the vitrification straw (aV) method. All groups were cultured in M-16 medium to determine the development and survivability for 24 h, respectively. Results shown that, the survivability of two different groups were significantly different (94.8% vs. 70.9%). Total cell number was not significantly different the non-frozen blastocyst ($99.7{\pm}12.4$) compared to the post-thaw blastocyst ($94.8{\pm}15.1$). From the 8-cell embryo, total cell number of frozen blastocysts were significantly lower than others groups ($74.7{\pm}14.6$, p<0.05). In the case of cell death analysis, the blastocysts from non-frozen and frozen-thawed 8-cell group were not different ($0.0{\pm}0.0$ vs. $1.9{\pm}3.1$, p>0.05). However, the apoptotic nuclei of blastocyst were significantly observed the frozen-thawed group ($5.4{\pm}4.4$) compared to non-frozen group (p<0.05). Therefore, this new method of embryos using in-straw dilution and direct transfer into other species would be more simple procedure of embryo transfer rather than step-wise dilution method and cryopreservation vessels, so we can be applied in animal as well as human embryo cryopreservation in further.

Effect of freezing on electrical properties and quality of thawed chicken breast meat

  • Wei, Ran;Wang, Peng;Han, Minyi;Chen, Tianhao;Xu, Xinglian;Zhou, Guanghong
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.4
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    • pp.569-575
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    • 2017
  • Objective: The objective of this research was to study the electrical properties and quality of frozen-thawed chicken breast meat and to investigate the relationship between these parameters at different times of frozen storage. Methods: Thawed samples of chicken breast muscles were evaluated after being kept in frozen storage at $-18^{\circ}C$ for different periods of time (1, 2, 3, 4, 5, 6, 7, and 8 months). Results: The results showed that water-holding capacity (WHC) and protein solubility decreased while thiobarbituric acid-reactive substances content increased with increasing storage time. The impedance module of samples decreased during 8-month frozen storage. Pearson correlation coefficients showed that the impedance change ratio (Q value) was significantly (p<0.05) related to pH, color, WHC, lipid oxidation and protein solubility, indicating a good relationship between the electrical properties and qualities of frozen-thawed chicken breast meat. Conclusion: Impedance measurement has a potential to assess the quality of frozen chicken meat combining with quality indices.

Unfrozen Water Content and Unconfined Compressive Strength of Frozen Soils according to Degree of Saturations and Silt Fractions (포화도와 실트 함량에 따른 동결토의 부동 수분량 및 일축압축강도 특성)

  • Kim, Sang Yeob;Hong, Won-Taek;Hong, Seung Seo;Baek, Yong;Lee, Jong-Sub
    • Journal of the Korean Geotechnical Society
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    • v.32 no.12
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    • pp.59-67
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    • 2016
  • The strength of frozen soils is affected by size and shape of particles, and the amount of ice and unfrozen water. The objective of this study is to characterize the unfrozen water content and the unconfined compressive strength of the frozen soils according to the degree of saturations and silt fractions. The specimens are mixtures of sand, silt, and water. The silt fractions (SF), which are the ratio of the silt weight ($W_{silt}$) to the sand weight ($W_{sand}$), are 10% and 30%. In addition, the degrees of the saturation are 5%, 10%, 15%, and 20%. The specimens are frozen under the temperature of $-10^{\circ}C$ conditions. The uniaxial compression tests are conducted for 24 hours, 48 hours, and 72 hours after freezing to determine proper freezing time. The freezing time of 24 hours is chosen because the unconfined compressive strengths of specimens after 24 hours freezing times are similar to each other. Furthermore, the unfrozen water content is monitored during freezing using the TDR system. The unfrozen water content increases with the increase of the silt fraction and degree of saturation. The unconfined compressive strength of the frozen soils exponentially increases with increasing the degree of saturation. This study shows that the amount of ice has more influence on the strength of the frozen soils than the amount of unfrozen water.