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Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Imaging Neuroreceptors in the Living Human Brain

  • Wagner Jr Henry N.;Dannals Robert F.;Frost J. James;Wong Dean F.;Ravert Hayden T.;Wilson Alan A.;Links Jonathan M.;Burns H. Donald;Kuhar Michael J.;Snyder Solomon H.
    • The Korean Journal of Nuclear Medicine
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    • v.18 no.2
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    • pp.17-23
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    • 1984
  • For nearly a century it has been known that chemical activity accompanies mental activity, but only recently has it been possible to begin to examine its exact nature. Positron-emitting radioactive tracers have made it possible to study the chemistry of the human mind in health and disease, using chiefly cyclotron-produced radionuclides, carbon-11, fluorine-18 and oxygen-15. It is now well established that measurable increases in regional cerebral blood flow, glucose and oxygen metabolism accompany the mental functions of perception, cognition, emotion and motion. On May 25, 1983 the first imaging of a neuroreceptor in the human brain was accomplished with carbon-11 methyl spiperone, a ligand that binds preferentially to dopamine-2 receptors, 80% of which are located in the caudate nucleus and putamen. Quantitative imaging of serotonin-2, opiate, benzodiazapine and muscarinic cholinergic receptors has subsequently been accomplished. In studies of normal men and women, it has been found that dopamine and serotonin receptor activity decreases dramatically with age, such a decrease being more pronounced in men than in women and greater in the case of dopamine receptors than serotonin-2 receptors. Preliminary studies in patients with neuropsychiatric disorders suggests that dopamine-2 receptor activity is diminished in the caudate nucleus of patients with Huntington's disease. Positron tomography permits quantitative assay of picomolar quantities of neuro-receptors within the living human brain. Studies of patients with Parkinson's disease, Alzheimer's disease, depression, anxiety, schizophrenia, acute and chronic pain states and drug addiction are now in progress. The growth of any scientific field is based on a paradigm or set of ideas that the community of scientists accepts. The unifying principle of nuclear medicine is the tracer principle applied to the study of human disease. Nineteen hundred and sixty-three was a landmark year in which technetium-99m and the Anger camera combined to move the field from its latent stage into a second stage characterized by exponential growth within the framework of the paradigm. The third stage, characterized by gradually declining growth, began in 1973. Faced with competing advances, such as computed tomography and ultrasonography, proponents and participants in the field of nuclear medicine began to search for greener pastures or to pursue narrow sub-specialties. Research became characterized by refinements of existing techniques. In 1983 nuclear medicine experienced what could be a profound change. A new paradigm was born when it was demonstrated that, despite their extremely low chemical concentrations, in the picomolar range, it was possible to image and quantify the distribution of receptors in the human body. Thus, nuclear medicine was able to move beyond physiology into biochemistry and pharmacology. Fundamental to the science of pharmacology is the concept that many drugs and endogenous substances, such as neurotransmitters, react with specific macromolecules that mediate their pharmacologic actions. Such receptors are usually identified in the study of excised tissues, cells or cell membranes, or in autoradiographic studies in animals. The first imaging and quantification of a neuroreceptor in a living human being was performed on May 25, 1983 and reported in the September 23, 1983 issue of SCIENCE. The study involved the development and use of carbon-11 N-methyl spiperone (NMSP), a drug with a high affinity for dopamine receptors. Since then, studies of dopamine and serotonin receptors have been carried out in over 100 normal persons or patients with various neuropsychiatric disorders. Exactly one year later, the first imaging of opitate receptors in a living human being was performed [1].

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Effect of cytokines in breast milk on infant during the first two-years of life (모유 내 사이토카인 함량과 2 년간 추적관찰 후 알레르기 발생)

  • Kim, Woo Kyung;Yoon, Hae-Sun
    • Clinical and Experimental Pediatrics
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    • v.50 no.9
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    • pp.912-918
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    • 2007
  • Purpose : Breast milk contains several components that provide specific immunity and affect the maturation of the infant's immune system. Allergic disease (AD), including atopic eczema, asthma, allergic rhinitis, and food allergy is characterized by an imbalance between cytokines produced by distinct T-helper cell subtypes. The aim of the study was to investigate the concentrations of cytokines and chemokines that were involved in allergic reactions in breast milk from allergic and nonallergic mothers and then analyse the effect of breastfeeding duration on the prevalence of allergic disease in the age of two. Methods : The breast milk samples were collected from mothers with AD (n=88) and without AD (n=47). Breast milk was collected at the second day (colostrum) and four weeks later (mature milk).The level of Interlukine (IL)-4, IL-5, IL-6, IL-8, IL-10, IL-13, $TGF-{\beta}1$, $TGF-{\beta}2$, RANTES in breast milk were determined by commercially available enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer's instructions. Results : Mothers with AD had a higher concentration of IL-8 in colostrum compared with those without AD (P=0.021). But, $TGF-{\beta}1$ and $TGF-{\beta}2$ were higher concentrated in colostrum of mother without AD (P=0.013, P=0.001). Whereas concentrations of other cytokines were not significantly different between the two groups. There was no association between levels of cytokines and chemokines in the breast milk and allergic development during the first 2 years of life in the infants. Conclusion : The higher concentration of $TGF-{\beta}1$ and $TGF-{\beta}2$ in colostrum from non-allergic mothers may explain the protective effect. But, the higher concentrations of IL-8 in colostrum from allergic mothers may in part explain the controversial results on the protective effect of breastfeeding against allergic diseases. We conclude that there is no convincing evidence form a relation between cytokines in breast milk and allergic diseases in infants. Longer follow-up are necessary to evaluate the effects of breast milk components on AD.

Growth of Radish Plug Seedlings as Influenced by Various Pre-planting Nitrogen Levels in Inert Media (기비로 혼합된 질소 수준이 무 플러그 묘 생장에 미치는 영향)

  • Sung, Jwa Kyung;Park, Myong Sun;Choi, Jong Myung
    • Journal of Bio-Environment Control
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    • v.26 no.3
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    • pp.201-207
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    • 2017
  • Objective of this research was to investigate the influence of pre-plant nitrogen levels in root media on plug seedling growth of radish cv. Soksungbommu. To achieve the research purpose, a root medium, the mixture of perlite, coir dust, and peatmoss (volume percentage of 30:35:35) was formulated and the N levels incorporated during mixing were controlled to 0, 100, 250, 500, 750, 1,000, and $1,500mg{\cdot}L^{-1}$. Then, the seeds were sown into 72-cell plug trays in which the root medium was packed. The measurements of growth and analysis of tissue and root media were conducted 2 and 4 weeks after sowing. Elevation of pre-plant N levels raised EC and turned down pH of root media. But, as seedling grew, the pH rose and EC get lowered in all treatments. The EC as well as $NH_4-N$ and $NO_3-N$ concentrations of root media declined gradually until week 2, but those declined sharply between weeks 2 to 4. The seedling growth 2 weeks after sowing showed quadratic response to pre-plant N levels with the highest growth in $250mg{\cdot}L^{-1}$ treatment and lagging growth in the treatments of lower or higher N levels than $250mg{\cdot}L^{-1}$. The seedling growth 4 weeks after sowing showed also quadratic response with the highest growth in $500mg{\cdot}L^{-1}$ treatment. The tissue N contents were get higher and those of K, Ca, and Mg were get lower as pre-plant N levels were elevated. Above results suggest that lower than $250mg{\cdot}L^{-1}$ in pre-plant N levels is optimistic for growth of plug seedling and avoidance of toxic injury in very young stage.

Association between polymorphisms in Interleukin-17 receptor A gene and childhood IgA nephropathy (IgA 신병증 환자에서 Interleukin-17 수용체 A 유전자의 단일염기다형성 연관성 연구)

  • Baek, Seung-Ah;Han, Won-Ho;Cho, Byoung-Soo;Kim, Sung-Do
    • Clinical and Experimental Pediatrics
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    • v.53 no.2
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    • pp.215-221
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    • 2010
  • Purpose : Interleukin-17 (IL-17) is produced by activated CD4+T cells and exhibits pleiotropic biological activity on various cell types. IL-17 was reported to be involved in the immunoregulatory response in IgA nephropathy (IgAN). Our aim was to investigate the association between single-nucleotide polymorphisms (SNPs) in IL-17 receptor A (IL-17RA) gene and childhood IgAN. Methods : We analyzed the SNPs in the IL-17RA in 156 children with biopsy-proven IgAN and 245 healthy controls. We divided the IgAN patients into 2 groups and compared them with respect to proteinuria (${\leq}4$ and >$4mg/m^2/h$, ${\leq}40$ and >$40mg/m^2/h$, respectively) and the presence of pathological levels of biomarkers of diseases such as interstitial fibrosis, tubular atrophy, or global sclerosis. Results : No difference was observed between the SNP genotypes rs2895332, rs1468488, and rs4819553 between IgAN patients and control subjects. In addition, no significant difference was observed between allele frequency of SNPs rs2895 332, rs1468488, and rs4819553 between patients in the early and advanced stage of the disease. However, significant difference was observed between the genotype of SNP rs2895332 between patients with proteinuria (>$4mg/m^2/h$) and those without proteinuria (codominant model OR 0.36, 95% CI 0.19-.66, P <0.001; dominant model OR 0.35, 95% CI 0.17-.69 P =0.002; recessive model OR 0.12, 95% CI 0.01-.06 P =0.025). Conclusion : Our results indicate that the SNP in IL-17RA (rs2895332) may be related to the development of proteinuria in IgAN patients.

Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionic Gonadotropin Receptor

  • Min, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.357-364
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Usefulness of $^{99m}Tc$-labeled RBC Scan and SPECT in the Diagnosis of Head and Neck Hemangiomas (두경부 혈관종 진단시 $^{99m}Tc$-RBC Scan and SPECT 검사의 유용성)

  • Oh, Shin-Hyun;Roh, Dong-Wook;Ahn, Sha-Ron;Park, Hoon-Hee;Lee, Seung-Jae;Kang, Chun-Goo;Kim, Jae-Sam;Lee, Chang-Ho
    • The Korean Journal of Nuclear Medicine Technology
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    • v.12 no.1
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    • pp.39-43
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    • 2008
  • Purpose: There are various methods to diagnose hemangioma, such as ultrasonography (US), computed tomography (CT), magnetic resonance imaging (MRI) and nuclear medicine. However, by development of SPECT imaging, the blood-pool scan using $^{99m}Tc$-labeled red blood cell has been used, because it was non-invasive and the most economical method. Therefore, in this study, we proposed that the usefulness of $^{99m}Tc$-RBC scan and SPECT of the head and neck to diagnose unlocated hemangiomas. Materials and Methods: $^{99m}Tc$-RBC scan and SPECT was performed on 6 patients with doubtful hemangioma (4 person, head; 1 person, neck; 1 person, another). We labeled radiopharmaceutical using modified in vivo method and then, centrifuged it to remove plasma. After a bolus injection of tracer, dynamic perfusion flow images were acquired. Then, anterior, posterior, both lateral static blood-pool images were obtained as early and 4 hours delayed. SPECT was progressed 64 projections per 30 seconds. Each image was interpreted by physicians, Nuclear medicine specialist, and technologist blinded to patient's data. Results: In 5 patients of all the radioactivity of doubtful site didn't change in flow images, but, in blood-pool, delayed and SPECT images, it was increased. So, it was a typical hemangioma finding. The size of lesion was over 2 cm, and it could discriminate as comparing to the delayed and SPECT imaging. On the other hand, in 1 patient, the radioactivity was increased in blood-pool images, but, not in delayed and SPECT images, so, it was proved no hemangioma. Conclusion: Using $^{99m}Tc$-RBC Scan and SPECT, we could diagnose the hemangiomas in head and neck, as well as, liver, more non-invasive, economical, and easy. Therefore, it considered that $^{99m}Tc$-RBC scan and SPECT would offer more useful information for diagnosis of hemangioma, rather than otherimaging such as US, CT, MRI.

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Physiological Activity and Nutritional Composition of Pleurotus Species (느타리속 버섯류의 영양성분 및 생리활성)

  • Um, Su-Na;Jin, Gyoung-Ean;Park, Kye-Won;Yu, Young-Bok;Park, Ki-Moon
    • Korean Journal of Food Science and Technology
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    • v.42 no.1
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    • pp.90-96
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    • 2010
  • In this study, the anti-oxidant, anti-tumorigenic, anti-hypertensive, anti-thrombic, anti-diabetic, and anti-inflammatory properties of 18 different species of genus Pleurotus were investigated. In addition, the amino acid, $\beta$-glucan, and polyphenol content were also measured. All species contained more than 20 mg% of polyphenol with the highest contents found in Pleurotus cornucopiae var. citrinopileatus (yellow pleurotus) ($39.13{\pm}0.82\;mg%$). The $\beta$-glucan contents was also the highest in yellow Pleurotus ($37.67{\pm}0.22%$) followed by Won-Hyeong1 (C, $28.75{\pm}0.61%$) and Jang-an PK (A, $27.95{\pm}0.33%$). The yellow Pleurotus exhibited the highest antioxidant activity as assessed by the DPPH scavenging rate with an $IC_{50}$ value of $2.94{\pm}0.44\;mg/mL$. Ethanol extracts from the yellow Pleurotus treated at 1% concentration showed cytotoxic activity up to 36.9% in the human embryonic kidney 293T cell lines. The yellow Pleurotus also showed the highest inhibitory effects on ACE activity ($60.52{\pm}0.2%$). Finally, the yellow Pleurotus exhibited anti-diabetic and anti-inflammatory properties as shown by inhibition of $\alpha$-amyloglucosidase activity ($50.5{\pm}0.8%$) and nitric oxide production ($68.4{\pm}0.3%$). Taken together, our data indicate the yellow pleurotus is a promising functional food ingredients.

Clinical Outcomes of Corrective Surgical Treatment for Esophageal Cancer (식도암의 외과적 근치 절제술에 대한 임상적 고찰)

  • Ryu Se Min;Jo Won Min;Mok Young Jae;Kim Hyun Koo;Cho Yang Hyun;Sohn Young-sang;Kim Hark Jei;Choi Young Ho
    • Journal of Chest Surgery
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    • v.38 no.2 s.247
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    • pp.157-163
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    • 2005
  • Background: Clinical outcomes of esophageal cancer have not been satisfactory in spite of the development of surgical skills and protocols of adjuvant therapy. We analyzed the results of corrective surgical patients for esophageal cancer from January 1992 to July 2002. Material and Method: Among 129 patients with esophageal cancer, this study was performed in 68 patients who received corrective surgery. The ratio of sex was 59 : 9 (male : female) and mean age was $61.07\pm7.36$ years old. Chief complaints of this patients were dysphagia, epigastric pain and weight loss, etc. The locations of esophageal cancer were 4 in upper esophagus, 36 in middle, 20 in lower, 8 in esophagogastric junction. 60 patients had squamous cell cancer and 7 had adenocarcinoma, and 1 had malignant melanoma. Five patients had neoadjuvant chemotherapy. Result: The postoperative stage I, IIA, IIB, III, IV patients were 7, 25, 12, 17 and 7, respectively. The conduit for replacement of esophagus were stomach (62 patients) and colon (6 patients). The neck anastomosis was performed in 28 patients and intrathoracic anastomosis in 40 patients. The technique of anastomosis were hand sewing method (44 patients) and stapling method (24 patients). One of the early complications was anastomosis leakage (3 patients) which had only radiologic leakage that recovered spontaneously. The anastomosis technique had no correlation with postoperative leakage, which stapling method (2 patients) and hand sewing method (1 patient). There were 3 respiratory failures, 6 pneumonia, 1 fulminant hepatitis, 1 bleeding and 1 sepsis. The 2 early postoperative deaths were fulminant hepatitis and sepsis. Among 68 patients, 23 patients had postoperative adjuvant therapy and 55 paitents were followed up. The follow up period was $23.73\pm22.18$ months ($1\~76$ month). There were 5 patients in stage I, 21 in stage 2A, 9 in stage IIB, 15 in stage III and 5 in stage IV. The 1, 3, 5 year survival rates of the patients who could be followed up completely was $58.43\pm6.5\%,\;35.48\pm7.5\%\;and\;18.81\pm7.7\%$, respectively. Statistical analysis showed that long-term survival difference was associated with a stage, T stage, and N stage (p<0.05) but not associated with histology, sex, anastomosis location, tumor location, and pre and postoperative adjuvant therapy. Conclusion: The early diagnosis, aggressive operative resection, and adequate postoperative treatment may have contributed to the observed increase in survival for esophageal cancer patients.