• Title/Summary/Keyword: mammary tissue

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Production of transgenic cattle by somatic cell nuclear transfer (SCNT) with the human granulocyte colony-stimulation factor (hG-CSF)

  • Carvalho, Bruno P.;Cunha, Andrielle T.M.;Silva, Bianca D.M.;Sousa, Regivaldo V.;Leme, Ligiane O.;Dode, Margot A.N.;Melo, Eduardo O.
    • Journal of Animal Science and Technology
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    • v.61 no.2
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    • pp.61-68
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    • 2019
  • The hG-CSF (human Granulocyte Colony-Stimulating Factor) is a growth and stimulation factor capable of inducing the proliferation of bone marrow cells, several types of leukocytes, among other hematopoietic tissue cells. hG-CSF is used in used to treat anomalies that reder a small number of circulating white blood cells, which may compromise the immune defenses of the affected person. For these reasons, the production of hG-CSF in a bioreactor system using the mammary gland of genetic modified animals is a possibility of adding value to the bovine genetic material and reducing the costs of hG-CSF production in pharmaceutical industry. In this study, we aimed the production of transgenic hG-CSF bovine through the lipofection of bovine primary fibroblasts with an hG-CSF expression cassette and cloning these fibroblasts by the somatic cell nuclear transfer (SCNT) technique. The bovine fibroblasts transfected with the hG-CSF cassette presented a stable insertion of this construct into their genome and were efficiently synchronized to G0/G1 cell cycle stage. The transgenic fibroblasts were cloned by SCNT and produced 103 transferred embryos and 2 pregnancies, one of which reached 7 months of gestation.

Retroperitoneal Suppurative Fistula Caused by Remnant Suture Material in a Poodle Dog

  • Kim, Keunyung;Lee, Chaeyeong;Kim, Minyeon;Choi, Hyeonjong;Hong, Jeongho;Kim, Hyoyeon;Park, Hyojin;Kim, Na-hyun;Lee, Dongbin;Lee, Jae-Hoon
    • Journal of Veterinary Clinics
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    • v.38 no.6
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    • pp.285-289
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    • 2021
  • An eight-year-old, 3.4 kg, spayed female poodle dog was referred to Gyeongsang National University Animal Medical Center with obstinate bilateral retroperitoneal subcutaneous abscess. The medical history revealed ovariohysterectomy (OHE), lipoma, and mammary gland tumor excision performed two years ago. The dog presented with left retroperitoneal subcutaneous abscess discharge five months back and was treated by abscess drainage and antibiotics in a local veterinary hospital, but a relapse was reported on discontinuing antibiotics. On physical examination, the dog was febrile (40℃) and both retroperitoneal masses showed typical inflammatory reaction and purulent discharge. On serum chemistry analysis, hyperproteinemia, hyperglobulinemia, and elevated levels of C-reactive protein were observed. Cytologic examination showed degenerative neutrophils and phagocytic macrophages. Radiological examination revealed encapsulated hypoechoic subcutaneous masses in bilateral abdominal flank and the left flank mass was connected with the caudal pole of left kidney through fistula. Based on the diagnostic examination, exploratory celiotomy was recommended and left nephrectomy, and resection of bilateral masses were performed. During procedure, fistula between caudal pole of left kidney and left subcutaneous abscess was detected and suture material was identified. Resected tissue was histologically examined and diagnosed as suppurative abscess caused by the suture material. This case report describes chronic suppurative foreign body reaction including caudal pole of left kidney and bilateral retroperitoneal subcutaneous masses induced by the suture material used in OHE two years ago and the necessity of computed tomography examination to identify character of mass and extent of surgical resection.

Two Cases of Supernumerary Nipple (과다 유두(Supernumerary Nipple) 2례)

  • Lee, Jeong-Woo;Yang, Jung-Dug;Lee, Jung-Hun;Choi, Kang-Young;Kim, Gui-Rak;Chung, Ho-Yun;Cho, Byung-Chae
    • Archives of Plastic Surgery
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    • v.37 no.5
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    • pp.695-698
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    • 2010
  • Purpose: Supernumerary nipple or polythelia is one of the developmental anomalies occurring at the embryonic stage and this anomaly usually arises from the milk line. While this atypical feature is determined during early developmental stage, it may not come out obviously or become troublesome until puberty or lactation. Moreover, sometimes it is confused with a pigmented nevus. Methods: Case 1, a 18-year-old woman with intramammary supernumerary breast consisted of another nipple with middle sized areola on the right lower breast was admitted for a $2.8{\times}3.1\;cm$-sized mass on the right breast which was starting appeared 1 year earlier. The preliminary cytological examination of the material obtained by needle aspiration biopsy from the mass was revealed by fibroadenoma with no malignant change. The patient had the surgical excision of the mass and accessory breast. Case 2, a 16 year-old woman admitted for intra-areolar polythelia of the left breast, even she doesn't have any family history of polythelia. Since she wanted surgical correction of her atypical nipple for aesthetic and psychological reasons, we reconstructed the areola using transposition flaps in an S-plasty design. Results: Case 1, the excised supernumerary nipple showed following histological features. In the superficial layer, an acanthotic and hyperpigmented epithelium with elongated rete ridges was found. In the dermis, there were follicles with hairs surrounded by hypertrophic sebaceous glands. In the deepest portion, abundant secretory glomerules and excretory ducts of apocrine gland type were observed. Case 2, follow-up visits 3 months after the procedure showed a satisfactory result with good shape and projection of the nipple. Conclusion: We report two cases of aberrant mammary tissue who underwent surgical correction, including complete breast (with nipple, areola, and glandular tissue) and intra-areolar polythelia according to the Kajava's classification, and the results were satisfactory.

Molecular characterization and expression pattern of a novel Keratin-associated protein 11.1 gene in the Liaoning cashmere goat (Capra hircus)

  • Jin, Mei;Cao, Qian;Wang, Ruilong;Piao, Jun;Zhao, Fengqin;Piao, Jing'ai
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.3
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    • pp.328-337
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    • 2017
  • Objective: An experiment was conducted to determine the relationship between the KAP11.1 and the regulation wool fineness. Methods: In previous work, we constructed a skin cDNA library and isolated a full-length cDNA clone termed KAP11.1. On this basis, we conducted a series of bioinformatics analysis. Tissue distribution of KAP11.1 mRNA was performed using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis. The expression of KAP11.1 mRNA in primary and secondary hair follicles was performed using real-time PCR (real-time polymerase chain reaction) analysis. The expression location of KAP11.1 mRNA in primary and secondary hair follicles was performed using in situ hybridization. Results: Bioinformatics analysis showed that KAP11.1 gene encodes a putative 158 amino acid protein that exhibited a high content of cysteine, serine, threonine, and valine and has a pubertal mammary gland) structural domain. Secondary structure prediction revealed a high proportion of random coils (76.73%). Semi-quantitative RT-PCR showed that KAP11.1 gene was expressed in heart, skin, and liver, but not expressed in spleen, lung and kidney. Real time PCR results showed that the expression of KAP11.1 has a higher expression in catagen than in anagen in the primary hair follicles. However, in the secondary hair follicles, KAP11.1 has a significantly higher expression in anagen than in catagen. Moreover, KAP11.1 gene has a strong expression in inner root sheath, hair matrix, and a lower expression in hair bulb. Conclusion: We conclude that KAP11.1 gene may play an important role in regulating the fiber diameter.

Transareolar-Perinipple Dual Pockets Breast Augmentation (횡유륜 유두주위절개를 통한 이중포켓 유방확대술)

  • Lee, Paik Kwon;Kim, Jee Hoon;Seo, Byung Chul;Oh, Deuk Young;Rhie, Jong Won;Ahn, Snag Tae
    • Archives of Plastic Surgery
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    • v.34 no.1
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    • pp.93-98
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    • 2007
  • Purpose: Many options are available for the incision and pocket selection in breast augmentation. Each method has its advantages and disadvantages. To leave an invisible operation scar and to achieve easier pocket dissection by the central location of the incision on the breast, we made a transareolar-perinipple incision. To overcome the disadvantages of the transareolar incision, originally advocated by Pitanguy in 1973, we modified the direction of incision line and dissection plane. Methods: To avoid the injury of 4th intercostal nerve responsible for nipple sensation, we made perinipple incision on the medial side of the nipple instead of trans-nipple incision and made the transareolar incision as 11-5 o'clock on the left side and 1-7 o'clock on the right side instead of 3-9 o'clock on both sides. To avoid the possible infection and breast feeding problem caused by the injury to the lactiferous duct, and the possible implant hernia caused by the incisions lying on a same plane of pocket dissection, we made a subcutaneous dissection just above the breast tissue medially down to the bottom of breast tissue and made a subglandular or subfascial pocket, which may avoid the injury of lactiferous duct and create different planes for skin incision and pocket dissection. Other advantages of the transareolar-perinipple incision include easier pocket dissection, less chance of hematoma, and as a result less postoperative pain because of the central location of the approach which allow finger dissection and meticulous bleeding control with direct vision, without any specialized instrument such as an endoscope or long mammary dissectors. As for pocket selection, we made dual pockets. We prefer subglandular or subfascial pocket. Also, we made a subpectoral pocket in the upper 1/4 of the pocket to add more volume on the upper part of the augmented breast, which can make aesthetically more desirable breasts in thin Asian women with small breasts. Possible disadvantages of our method are subclinical infection and scar widening, which could be overcome by meticulous operation techniques, antibiotic therapy, and intradermal tattooing. Results: From September, 2003 to August, 2005, 12 patients underwent breast augmentation using round smooth surface saline implants by our method. During the mean follow-up period of 13 months, there were no complications such as infection, hematoma, capsular contracture, and sensory change of nipple, and results were satisfactory. Conclusion: We suggest breast augmentation via transareolar-perinipple incision and dual pockets(subpectoral-subglandular or subfascial) as a valuable method in thin oriental women with small breasts.

Effect of Gamijipaesan Extracts against Mastitis Induced by Staphylococcus aureus Infection in a Rat Model through Anti-inflammatory and Antibacterial Effects (가미지패산(加味芷貝散)의 포도상구균 감염 유방염에 대한 항균활성 및 항염 효과)

  • Kwon, Ji-Myung;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.26 no.1
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    • pp.1-24
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    • 2013
  • Objectives: The object of this study was to observe the protective effect of Gamijipaesan aqueous extracts(GJS), which has been traditionally used in Korean medicine in obstetrics & gynecological fields as anti-infectious and anti-inflammatory agents, against mastitis induced by Staphylococcus aureus infection in a rat model through antibacterial, antiinflammatory, immunomodulatory, and anti-oxidant effects. Methods: Antibacterial activities of GJS against S. aureus were detected using standard agar microdilution methods, with the effects on the bacterial invasion and intracellular killing of individual test materials in human mammary gland carcinoma cell(MCF-7) and murine macrophages(Raw 264.7) at MIC1/2, MIC and MIC2 concentration levels. In addition, the effects on the cell viability, nitric oxide(NO), tumor necrosis factor(TNF)-${\alpha}$ and interleukin (IL)-6 productions of LPS activated Raw 264.7 cells. The changes on the mammary tissue viable bacterial numbers, myeloperoxidae(MPO), inducible nitric oxide synthetase(iNOS), TNF-${\alpha}$ and IL-6 contents were observed in the S. aureus in vivo intramammary infectious rat model. The anti-bacterial and anti-inflammatory effects were compared with ciprofloxacin and piroxicam, respectively in the present study. Results: MIC of GJS and ciprofloxacin against S. aureus were detected as $0.860{\pm}0.428$ (0.391-1.563) mg/ml and $0.371{\pm}0.262$(0.098-0.782) ${\mu}g/ml$, respectively. In addition, GJS and ciprofloxacin were also showed marked dosage-dependent inhibition of the both bacterial invasion and intracellular killing assays using MCF-7 and Raw 264.7 cells at MIC1/2, MIC and $MIC{\times}2$ concentrations, respectively. $ED_{50}$ against LPS-induced cell viabilities and NO, TNF-${\alpha}$ and IL-6 releases of GJS were detected as 0.72, 0.04, 0.08 and 0.11 mg/ml, and as 19.04, 4.18, 5.37 and 4.27 ${\mu}g/ml$ in piroxicam, respectively. 250 and 500 mg/kg of GJS also inhibit the intramammary bacterial growth, MPO, iNOS, TNF-${\alpha}$ and IL-6 contents in S. aureus in vivo intramammary infected rats, respectively. GJS 500 mg/kg showed quite similar antibacterial and anti-infectious effects as compared with ciprofloxacin 40 mg/kg and also showed similar anti-inflammatory effects as piroxicam 10 mg/kg, in S. aureus in vivo intramammary infectious models. Conclusions: The results obtained in this study suggest that over 250 mg/kg of GJS showed favorable anti-infectious effects against S. aureus infection in a rat model through their antibacterial, anti-inflammatory, immunomodulatory and anti-oxidant effects and therefore expected that GJS can be used as alternative therapies, having both anti-inflammatory and anti-infectious activities. However, more detail mechanism studies should be conducted in future with the efficacy tests of individual herbal composition of GJS and the screening of the biological active compounds in individual herbs. In the present study, GJS 500 mg/kg showed quite similar anti-infectious effects were detected as compared with ciprofloxacin 40 mg/kg treated rats, and also GJS shows quite similar anti-inflammatory effects as compared with piroxicam 10 mg/kg in S. aureus in vivo intramammary infectious rats, but ciprofloxacin did not showed any anti-inflammatory effects, and piroxicam did not showed anti-infectious effects in this study.

Effects of the cis-Acting Element in the 3' End of Porcine $\beta$-Casein Gene on the Expression in Mammary Epithelial Cells (돼지 $\beta$-Casein 유전자의 3' 말단 부위의 cis-Acting Element가 유선 상피 세포내의 발현에 미치는 영향)

  • Lee, Hwi-Cheul;Kim, Byoung-Ju;Byun, Sung-June;Lee, Seung-Hoon;Kim, Min-Ji;Chung, Hee Kyoung;Lee, Hyun-Gi;Jo, Su-Jin;Chang, Won-Kyong;Park, Jin-Ki;Lee, Poong-Yeon
    • Reproductive and Developmental Biology
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    • v.32 no.3
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    • pp.153-158
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    • 2008
  • Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.

Maspin Expression and Its Clinical Significance in Non-Small Cell Lung Cancer (비소세포폐암에서 Maspin의 발현과 임상적 의의)

  • Yoon, Seong-Hoon;Kim, Won-Jin;Shin, Kyung-Hwa;Kim, Mi-Hyun;Cho, Woo-Hyun;Kim, Ki-Uk;Park, Hye-Kyung;Jeon, Doo-Soo;Kim, Yun-Seong;Lee, Chang-Hun;Lee, Min-Ki;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.70 no.2
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    • pp.132-138
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    • 2011
  • Background: Maspin (mammary serine protease inhibitor) is a member of the serpin superfamily. A few studies have examined the role of maspin in tumor suppression of non-small cell lung cancer (NSCLC); however, its role in the development and progression of NSCLC still remains controversial. We evaluated the immunohistochemical expression of maspin in order to elucidate its clinical significance in NSCLC. Methods: We analyzed 145 patients with pathologically confirmed NSCLC, including 66 cases of squamous cell carcinomas (SCCs) and 79 cases of adenocarcinomas (ADCs). We performed a immuno-histochemical stain with maspin and PCNA (proliferating cell nuclear antigen) using tissue microarray blocks. Results: There were 108 men and 37 women in the study population. The mean age of patients in the study was 63.7 years (range, 40.0~82.0; median, 65.0). The proportion of maspin expression was significantly higher in SCCs (52/66, 78.8%; p<0.01) than in ADCs (17/79, 21.5%; p<0.01). Maspin expression was not associated with PCNA (p=0.828), lymph node involvement (p=0.483), or tumor stage (p=0.216), but showed correlation with well-to-moderate tumor differentiation (p=0.012). There was no observed correlation between maspin expression and survival with NSCLC (p=0.218). Conclusion: The present study suggests that maspin expression was significantly higher in SCCs than in ADCs and was associated with low histological grade. However, maspin expression was not an independent factor to predict a prognosis in NSCLC.

Efficient Production of Cloned Bovine Embryos from Transformed Somatic Cells (형질전환 체세포로부터 소 복제수정란의 효율적인 생산)

  • Wee G.;B. H Sohn;Park, J. S.;D. B. Koo;Lee, K. K.;Y. M. Han
    • Korean Journal of Animal Reproduction
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    • v.27 no.1
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    • pp.25-34
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    • 2003
  • Human thrombopoietin (hTPO) is a cytokine that plays a central role in megakaryopoiesis. To direct hTPO expression in the mammary gland, an expression vector was constructed by combining the promoter of bovine beta-casein gene, cDNA of hTPO and neomycin resistance gene (pBT-L neo). Fibroblast cells derived from cow's ear skin tissue were transfected with the expression vector (pBT-L neo) using Lipofectamine. Transfected cells resistant to G418 trea?nt were cultured to form the colonies for more than 2 weeks. The transformed colonies identified by PCR were further expanded prior to nuclear transfer. Reconstructed oocytes with transformed cells were electrofused, activated using calcium ionophore and 6-DMAP, and cultured in vitro for 7 days. Of 35 cell colonies analyzed by PCR, 29 colonies (82.9%) were positive for the hTPO gene. Cleavage and developmental rates to the blastocyst stage of reconstructed embryos with the transformed cells were 65.1% and 23.8%, respectively Of 29 blastocysts that developed from reconstructed embryos with the transformed cells, 27 embryos (93.1%) were transgenic. These results indicate that transgenic bovine embryos can be efficiently produced by somatic cell nuclear transfer using transformed cells.

The Effect of Vitamin A Derivatives on the Activity of Drug-metabolizing Enzyme in Rat Liver (Vitamin A 유도체로 인한 간의 약물대사효소 변동)

  • Lee, H.W.;Ryu, K.Z.;Ro, J.Y.;Hong, S.S.
    • The Korean Journal of Pharmacology
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    • v.18 no.1
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    • pp.65-72
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    • 1982
  • It has been known that retinoids are intrinsically of critical importance for control of premalignant epithelial cell differentiation. In the absence of retinoids, normal cellular differentiation and growth does not occur in epithelia such as those of trachea and bronchi. Furthermore, it was also reported that retinoid deficiency enhanced susceptibility to chemical carcinogenesis in the respiratory system, in the bladder, and in the colon of the experimental animal. In 1974, Bollag examined the effects of synthetic retinoids in prevention of development of cancer and demonstrated synthetic retinoids to have more favorable therapeutic index than retinoic acid for causing regression of skin papilloma in mice. Therefore, it was assumed that this anticarcinogenic effect of vitamin A derivatives could be due to modification of the metabolism of the carcinogenic polycyclic hydrocarbon, which must first be activated to exert their effect. Hill and Shih reported that vitamin A compounds and analogs had inhibitory effect on drug metabolizing enzyme from liver and lung tissue of mouse and hamster. Lucy suggested that the chemoprevention effect of vitamin A derivatives is due to reaction with molecular oxygen, and it is possible that inhibition of hydroxybenzpyrene formation is a result of this property. On the other hand, butylated hydroxytoluene which is a potent antioxidant strongly inhibited the formation of mammary tumor induced by dimethylbenranthracene. Also, it was observed that this antioxidant inhibited cancer induction in rats by N-2-fluo-renylacetamide. The purpose of this experiment was to investigate the effect of vitamin A derivatives such as retinoic acid and retinoid on drug-metabolizing enzyme and to determine whether riboflavin tetrabutylate or vitamin E could prevent of modify any changes induced by vitamin A delivatives in the rats. The results obtained were as followings. 1) Body weight was significantly reduced by retinoic acid, but not by retinoid. 2) Retinoic acid markedly increased liver weight while retincid showed no effect on liver weight. Treatment of riboflavin tetrabutylate did not affect retinoic acid-induced change in both body weight and liver weight. 3) Both retinoic acid and retinoid remarkably decreased the activity of aminopyrine demethylase. Pretreatment of riboflavin tetrabutylate, however, prevented inhibitory effect of retinoic acid on the enzyme activity. 4) No significant effect of vitamin E on aminopyrine demethylase was observed in both groups treated with retinoic acid and retinoid.

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