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Biomedical subjects

Y Giudicelli

Publications and source records attributed to Y Giudicelli.

At least 37 records · Page 2Linked to original sources

Insulinemia and pancreatic alpha2-adrenoceptors in salt-sensitive (SBH) and salt-resistant (SBN) Sabra rats: effect of high salt diet.

Studies in humans have suggested that hyperinsulinemia might play an important role in salt sensitivity and in the later development of high blood pressure. This possibility has been tested in this study on Sabra rats, an animal model of salt-induced hypertension. Salt-sensitive (SBH) and salt-resistant (SBN) Sabra rats have been submitted to either a normal (0.2% NaCl) or a high salt (8% NaCl) diet for 6 weeks. Comparisons of blood pressure, basal glucose and insulin levels, and insulin response to glucose overload (1 g/kg) have been made. As pancreatic alpha2-adrenergic receptors are implicated in the control of insulin release, their densities have been determined on plasma membranes by saturation studies with [3H]-RX-821002 as the specific radioligand. Under normal diet, blood pressures were respectively 133 +/- 9 and 108 +/- 10 mm Hg (n = 6) in SBH and SBN. Basal glucose and insulin levels and insulin response to glucose overload were found to be significantly higher in SBH than in SBN. In contrast, alpha2-adrenergic receptor densities were lower (P < .001) in SBH when compared to SBN. High salt diet increased (P < .01) blood pressure, decreased basal glucose (P < .01) and insulin (P < .001) levels only in SBH. However, when compared to SBN the insulin response to glucose overload was maintained higher in SBH. Alpha2-adrenergic receptor densities and difference between SBH and SBN did not differ from those found in normal diet. In conclusion, the salt-induced high blood pressure of salt-sensitive Sabra rats is not associated with hyperinsulinemia and insulin resistance. Indeed, an improvement in the insulin sensitivity appears to be induced by either a high salt diet or high blood pressure.

Animals↗

Androgen receptors in human preadipocytes and adipocytes: regional specificities and regulation by sex steroids.

Various clinical and epidemiological evidence strongly suggests a major role for sex steroid hormones in the determination of anatomical specificities of fat distribution in human. To date, no studies have examined the possible presence of androgen receptors (AR) in human adipocytes and preadipocytes. We have studied AR in preadipocytes from various anatomical locations (intra-abdominal and subcutaneous) in middle-aged men and women during the proliferation and differentiation processes (adipogenesis). Androgen binding sites quantified by [3H]R-1881-specific binding in whole cell extracts were twofold higher in intra-abdominal than in subcutaneous preadipocytes but identical for the same fat depots in men and women. Western blot analysis revealed 1) the presence of AR in the nuclear and cytosolic fractions of human preadipocytes, 2) a decrease of AR expression during adipogenesis, and 3) an upregulation of AR by androgens in vitro. RT-PCR experiments showed the presence of AR mRNA in human preadipocytes and adipocytes and also the regional specificity of AR distribution. However, AR mRNA expression was found to increase during adipogenesis. The same results were observed in rat preadipocytes. In conclusion, this study clearly demonstrates the presence of AR in human preadipocytes and adipocytes and suggests that androgens may contribute, through regulation of their own receptors, to the control of adipose tissue development.

Abdomen↗

Mutant alpha-subunit of the G protein G12 activates proliferation and inhibits differentiation of 3T3-F442A preadipocytes.

We studied the G protein alpha-subunit Galpha12 in various tissues and cell lines. Significant amounts of Galpha12 were detected by immunoblots in liver, chromaffin cells, RINm5F cells, 3T3-F442A cells, and preadipocytes, but not in adipocytes, sperm, kidney, NB2A cells, or brain. To study the role of Galpha12 in adipose tissue differentiation, the preadipocyte cell line 3T3-F442A was transfected with wild-type Galpha12 or a constitutively activated mutant of Galpha12. Stable expression of the activated mutant of Galpha12 stimulated cell growth and inhibited preadipocyte differentiation. In contrast, wild-type Galpha12 overexpression inhibited preadipocyte differentiation, without any effect on cell proliferation. The role of Galpah12 on the Raf/MEK/mitogen-activating protein kinase (MAPK) cascade was studied. In confluent preadipocytes, expression of the activated mutant of Galpha12 induced an increase in B-Raf expression, but no change in MAPK activity. Differentiation was associated with a decrease in MAPK activity in control 3T3-F442A cells. Wild-type Galpha12 overexpression prevented the decrease in MAPK activity and induced MEK1, but not B-Raf, expression. Moreover, the activated mutant of Galpha12 induced an increase in MAPK activity and in the expression of both MEK1 and B-Raf. These data indicate that the activated mutant of Galpha12 stimulates the proliferation of 3T3-F442A preadipocytes, possibly through an increase in B-Raf expression, independently of the MEK/MAPK pathway, but prevents differentiation, probably through an increase in MEK1 expression and MAPK activity.

Adipocytes↗

Congenital lumbosacral lipomas.

Congenital lumbosacral lipomas can be responsible for progressive defects. The general feeling is that tethering of roots, filum, or cord probably explains this evolution, and that untethering of these structures could prevent late deterioration. Like the vast majority of neurosurgeons, we too have routinely and systematically operated on lumbosacral lipomas, even in the absence of neurological deficits. This policy stemmed from our belief that spontaneous neurological deterioration was frequent, recovery from preoperative deficits rare, and surgery both efficient and benign in nature. After 22 years of experience, we felt that it was necessary to review our series of 291 lipomas (38 lipomas of the filum and 253 of the conus) operated on from 1972 to 1994. To reassess the value of prophylactic surgery, we attempted an accurate evaluation of (1) the risk of pathology, (2) the risks involved in surgery, (3) the postoperative outcome with respect to preoperative deficits, and (4) the postoperative outcome in asymptomatic patients at 1 year and at maximum follow-up. Special attention was paid to 93 patients whose postoperative follow-up was more than 5 years (average 8.7, median 8, range 5-23 years). Of these 93 patients, 39 were asymptomatic preoperatively (7 with lipoma of the filum and 32 with lipoma of the conus). Lipomas of the filum and of the conus are entirely different lesions and were studied separately. In 6 cases prenatal diagnosis had been possible. The mean age at surgery was 6.4 years. Low back skin stigmata were present in 89.4% of cases. Preoperative neurological deficits existed in 57% of the patients and were congenital in 22%. Clinical signs and symptoms recorded were pain in 13.3% of the patients and/or neurological deficits affecting sphincter (52%), motor (27.6%) and sensory (22.4%) functions. Deficits were progressive in 22.4% of cases, slowly progressive in 58.8% of these and rapidly progressive in the remaining 41.2%. In 36 patients (13.2%) the lipomas were seen to grow either subcutaneously or intraspinally. Among these patients, 21 were infants, 2 were obese adolescents, and 10 were pregnant women. The metabolism of the fat within the lipomas was studied in 11 patients and found to be similar to that at other sites. Lipomas were associated with various other malformations, either intra- or extraspinal. These associated anomalies were rare in the case of lipomatous filum (5.2%) but frequent with lipomas of the conus, except for intracranial malformations (3.6%). Therapeutic objectives were spinal cord untethering and decompression, sparing of functional neural tissue and prevention of retethering. Procedures used to achieve these goals were subtotal removal of the lipoma, intraoperative monitoring, duroplasty, and sometimes closure of the placode. Histologically, lipomas consisted of normal mature fat. However, 77% of them also included a wide variety of other tissues, originating from ectoderm, mesoderm, or entoderm. This indicates that lipomas are either simple or complex teratomas. The results of the study are as follows. (1) Surgery was easy and safe when performed for treatment of lipomas of the filum (no complications), but difficult and hazardous in the case of lipomas of the conus (20% local, 3.9% neurological complications). (2) All types of deficit could be improved by surgery, which was beneficial in all cases of lipoma of the filum and 50% of cases of lipoma of the conus. (3) In asymptomatic patients long-term surgical results depended on the anatomical type of the lipoma. They were excellent in lipomas of the filum. In lipomas of the conus they were good in the short term but eroded with time. At more than 5 years of follow-up only 53.1% of the patients were still free of symptoms. (4) Reoperations were performed in 16 patients (5.5%), 5 (31.2%) of whom improved postoperatively, while in 7 (43.7%) progression stopped, in 3 (18.7%) deterioration continued and in 1 (6.2%) the condition was wor

Adolescent↗

Evaluating the risk of preterm delivery: a comparison of fetal fibronectin and transvaginal ultrasonographic measurement of cervical length.

OBJECTIVE: Our purpose was to compare the predictive values for preterm delivery of fetal fibronectin and cervical length measured by transvaginal ultrasonography and to determine whether performing both tests improves their separate predictive values. STUDY DESIGN: This prospective blinded study performed both tests on 76 patients hospitalized with signs of premature labor between 24 and 34 weeks of gestation. The outcome measure was delivery before 37 weeks' gestation. RESULTS: The rate of preterm bith was 26.3% (20/76). The predictive values of fetal fibronectin and of a cervical length of < or = 26 mm, considered separately, were approximately equal, and the negative predictive value of each was excellent (86.6% and 89.1%, respectively). This value improved slightly when positive fetal fibronectin, a cervical length < or = 26 mm, or both defined abnormality (negative predictive value 94.4%). The positive predictive values, although less helpful, were still useful (45.2% and 50.0%, respectively). Combining both indicators did not noticeably improve the positive predictive value (52.4%). The risk of preterm delivery for a patient with a positive fetal fibronectin level and a short cervix was high (odds ratio 13.9, 95% confidence interval 3.7 to 52.2). CONCLUSION: Fetal fibronectin and cervical length are approximately equivalent in their ability to distinguish between patients at high and low risk for preterm delivery. For physicians equipped to perform transvaginal ultrasonography, however, the additional information about the fibronectin level provides only slight benefits.

Adult↗

Control of rat preadipocyte adipose conversion by ovarian status: regional specificity and possible involvement of the mitogen-activated protein kinase-dependent and c-fos signaling pathways.

As ovariectomy induces obesity in rats, we have investigated the influence of ovariectomy and hormone replacement on the proliferation and differentiation capacities of rat cultured preadipocytes removed from different fat depots (femoral sc, parametrial, and perirenal). Ovariectomy induced increased proliferation and differentiation as well as high mitogen-activated protein (MAP) kinase activity and c-fos protein induction in both confluent and differentiated preadipocytes from perirenal fat depots. In parametrial preadipocytes, ovariectomy also increased proliferation and c-fos protein induction, but failed to alter the capacities of these cells to differentiate. Treatment of ovariectomized rats with estradiol and progesterone reversed the promoting effect of ovariectomy on proliferation, differentiation, and c-fos induction in perirenal preadipocytes, but not the MAP kinase activation observed during the proliferative phase. This treatment also reversed the promoting effect of ovariectomy on proliferation and c-fos induction seen in confluent parametrial preadipocytes. In contrast, sc preadipocytes were totally insensitive to ovarian status in terms of proliferation and differentiation capacities, MAP kinase activity, and c-fos induction. This study demonstrates that adipogenesis is site-specifically controlled by the ovarian status in the rat. It also suggests that ovariectomy-induced obesity (mainly abdominal) could be related to changes in some of the signaling pathways controlling adipogenesis in intraabdominal preadipocytes.

Adipose Tissue↗

Site-related specificities of the control by androgenic status of adipogenesis and mitogen-activated protein kinase cascade/c-fos signaling pathways in rat preadipocytes.

In rats, castration induces a complete defective adipose conversion of preadipocytes from the epididymal fat depots (Lacasa, D., B. Agli, D. Noynarol, and Y. Giudicelli, 1995, Endocrine 3: 789-793). The aim of this study was to establish the eventual site-specificity of this effect as well as the mechanisms involved. Therefore, the influence of androgenic status on the Fos protein induction and the Raf/mitogen-activated protein (MAP) kinase kinase (MEK)MAP cascade, which are all required for adipose conversion of preadipocytes, was compared in proliferating and differentiated preadipocytes from femoral sc and deep intraabdominal (epididymal and perirenal) fat depots. In epididymal and perirenal proliferating preadipocytes, increased proliferation due to castration is associated with increased MAP kinase activity. However, higher immunoreactive levels of the upstream activators of MAP kinase, Raf-1 and MEK, were observed only in epididymal cells. Moreover, in vivo testosterone treatment corrected the effects of castration on Raf-1 but not on MEK and MAP kinase. MAP kinase activity was decreased during the course of adipogenesis. In differentiated cells, MAP kinase activity showed variations according to the anatomical origin of preadipocytes but not to the androgenic status. In contrast, MEK and Raf-1 immunoreactive levels were both sensitive to androgenic status but were differently affected depending on cell origin. Finally, the defective adipogenesis seen in epididymal preadipocytes from castrated rats was associated with reduced Fos protein induction in these cells, an alteration which was partly corrected by testosterone-treatment. Taken together, these results suggest that androgenic status affects adipogenesis from deep intraabdominal preadipocytes through alterations of some components of the MAP kinase cascade/Fos signaling pathways.

Adipocytes↗

Fibrinogen Poissy I: a new case of the A alpha Arg 16His fibrinogen variant.

A fibrinogen variant was identified in a patient with disseminated intravascular coagulation and in one member of her family. Coagulation studies showed marked prolongation of both the thrombin and reptilase times and discrepancy was noted between the levels of plasma fibrinogen, determined by a kinetic vs immunological determination or light scattering assay. Studies on purified fibrinogen revealed an impaired release of fibrinopeptides by thrombin. DNA sequencing revealed a heterozygous A to G point mutation in exon 2 of the A alpha chain, which substituted Arg for His at position 16. This mutation creates a Nla III cleavage site which was used to confirm the mutation.

Adult↗

White adipose tissue nitric oxide synthase: a potential source for NO production.

Nitric oxide synthase (NOS) activity was detected in soluble and membranous fractions of adipose tissue homogenates of control rats. After LPS-treatment, this activity was (i) markedly increased (about 10-fold) in both fractions, (ii) unaltered after dexamethasone pretreatment, (iii) partly calcium-calmodulin sensitive, and (iv) almost entirely accounted by the NOS activity found in isolated adipocytes. In adipose tissue homogenates from control rats, Western blot analysis demonstrated the presence of the endothelial (eNOS) isoform in the membranous fraction of control rats and of the inducible (iNOS) isoform in the soluble and membranous fractions. After LPS treatment, the amount of immunoreactive iNOS protein was dramatically increased, suggesting that adipose tissue is an important site of NO production during the endotoxic shock.

Adipose Tissue↗

Site-related differences in G-protein alpha subunit expression during adipogenesis in vitro: possible key role for Gq/11 alpha in the control of preadipocyte differentiation.

G protein alpha subunits were compared by immunoblotting in preadipocytes and adipocytes from rat subcutaneous and epididymal adipose tissue at three steps of adipogenesis. The most striking difference concerned the Gq/11 alpha subunits whose expression decreased during preadipocyte differentiation in subcutaneous cells while it remained constant in epididymal cells. The PKC inhibitor CGP 41251 increased glycerol 3-phosphate dehydrogenase activity (EC 1.1.1.8), a late marker of differentiation, in epididymal preadipocytes but not in subcutaneous cells. There was no difference in the proliferation capacities between subcutaneous and epididymal preadipocytes: in both cells, CGP 41251 led to the same decrease in [(3)H]thymidine incorporation and, at confluence, the amounts of Gq/11 alpha subunits were equivalent. No major site-related difference was found in the amounts of Gs and Gi alpha during adipogenesis. Thus, compared to epididymal preadipocytes, the higher capacity of subcutaneous preadipocyte to differentiate seems to be correlated with the decrease in Gq/11 alpha expression and the decreased Gq/11 mediated PKC activation.

Adipocytes↗

G proteins in adipocytes and preadipocytes: characterization, subcellular distribution, and potential roles for Gi2 and/or Gi3 in the control of cell proliferation.

Guanosine triphosphate (GTP)-binding protein subunits were studied by immunoblot analysis in particulate fractions from mature adipocytes, confluent preadipocytes, and in vitro-differentiated preadipocytes. Mature adipocytes express Gi alpha 1, Gi alpha 2, Gi alpha 3, Go alpha, Gq/11 alpha, G13 alpha and the long and short isoforms of Gs alpha, but no Gz alpha or G12 alpha. Confluent and differentiated preadipocytes differ in having a higher content of Gi alpha 3 and G13 alpha and expressing G12 alpha. In contrast, they lack Gi alpha 1, Go alpha, and the short from of Gs alpha. The G-protein alpha subunits Gi alpha 2, Gs alpha (long isoform), and Gq/11 alpha, and G-protein beta subunits were unchanged throughout the differentiation process. By immunoblot and indirect immunofluorescence studies on confluent preadipocytes, we showed that Gi alpha 2 is present in the endoplasmic reticulum and marginally in plasma membranes and nuclei. In contrast, antibodies to Gi alpha 3 stained the Golgi apparatus. The role of G proteins on preadipocyte proliferation was studied using Bordetella pertussis toxin. Exposure of growing cells to this toxin in the presence of fetal calf serum (FCS) decreased [3H]thymidine incorporation by 40% and induced a 40% increase in doubling time. This resulted in a 30% decrease in cell number per well after 48 h. These effects of B. pertussis toxin did not appear to be related to an increase in cyclic adenosine monophosphate (cAMP) concentration, because forskolin had the opposite effect on cell proliferation. Finally, B. pertussis toxin prevented serum-induced Raf1 association to the plasma membrane, possibly by disrupting FCS-induced G beta gamma effects on the Ras/Raf1 pathway. Since Go alpha and Gi alpha 1 subunits were absent in preadipocytes, we conclude that Gi2 and/or Gi3 proteins transduce some mitogenic signals of FCS through release of G beta gamma subunits. The subcellular distribution of Gi alpha 2 and Gi alpha 3 suggests that part of their functions result from interactions with components other than the plasma membrane.

Adipocytes↗

Activated protein C sensitivity ratio in pregnant women at delivery.

We studied activated protein C sensitivity ratio (APC-SR), factors V and VIII activity and von Willebrand antigen in control women, women using oral contraceptives, and pregnant women at delivery. The mean APC-SR of 2.4 in pregnant women was significantly lower than the mean APC-SR value of 3.5 for both the other groups and 45% of pregnant women had a ratio below the 5th percentile of the control group. None of them carried the R506-->Q mutation. This decreased ratio at delivery appeared to be connected, at least in part, with increased VIII activity. Thus, APC-SR at delivery should not be used to detect APC resistance.

Adult↗

Role of factor VIII on activated protein C resistance ratio in inflammatory diseases.

Activated protein C resistance ratio (APC-Rr), factor VIIIC (FVIIIC) and plasma fibrinogen levels were studied in patients with inflammatory disease. The patient mean APC-Rr was significantly lower than in the control group. This decreased ratio in inflammatory diseases appeared to be connected with increased FVIIIC. Moreover, supplementation of plasmas with purified factor VIII decreased the APC-Rr in plasma from both groups, and suppressed the difference between groups. These data suggest that factor VIIIa and factor Va compete for protein C-catalysed cleavage. Ratios were identical in both groups when FVIIIC level was lowered by dilution in factor V deficient plasma.

Binding, Competitive↗

[Rational use of fetal fibronectin in the evaluation of premature labor risk].

OBJECTIVE: To better determined the usefulness of fetal fibronectin assay to identify patients at risk of premature delivery. SUBJECTS: 155 primiparous or multiparous patients presenting with single or twin pregnancies between 24 and 34 weeks and divided into 3 groups: high risk (70 patients), medium risk (35 patients), and low risk (50 patients) of premature delivery. MAIN OUTCOME MEASURES: The occurrence of delivery in the 21 days following sampling and/or a premature delivery. RESULTS: Among the 50 patients in the low risk group, there was one false positive and no premature delivery. Among the 35 patients in the medium risk group, there was no single positive fetal fibronectin test and no premature delivery. Among the 70 patients in the high risk group, 14 had a premature rupture of the membranes with very strongly positive fetal fibronectin test and all were delivery prematurely, 11 within a period of less than 21 days. The remaining 56 patients presented a threat of premature delivery without rupture of membranes; 20 gave birth prematurely (prevalence: 35.7%). In patients with a negative fetal fibronectin test, those with positive fetal fibronectin test were significantly more likely to experience preterm birth (odds ration: 12; 95% confidence interval: 3.4 to 42.1; p = 0.001) or to deliver within 21 days (odds ratio: 29.9; 95% confidence interval: 13.3 to 243; p < 0.001). CONCLUSION: Measurement of fetal fibronectin in cervico-vaginal secretions enabled us to define an authentic sub-group at high risk of premature delivery among patients presenting uterine contractions and changes in the cervix. In contrast, measurement of fetal fibronectin in cervico-vaginal secretions of patients with low and medium risk is not suitable, due to the low rate of premature delivery in these groups and the significant increase in the cost of pregnancy monitoring.

Adult↗

Zeta PKC in rat preadipocytes: modulation by insulin and serum mitogenic factors and possible role in adipogenesis.

Several PKC isoforms belonging to the three PKCs' subfamilies cPKC alpha and beta isoforms, nPKC epsilon isoform and aPKC zeta isoform were detected by Western blot in rat preadipocytes. zeta PKC which appears involved in proliferation and differentiation of some cellular types was shown to display variations according to the preadipocyte anatomical origin and stage of differentiation. A rapid increase in zeta PKC in the cytosolic compartment and translocation into the nucleus were induced by mitogenic factors in proliferating preadipocytes and by insulin in differentiating preadipocytes. These findings suggest that zeta PKC could be involved i) in the post-receptor signaling pathway of serum mitogenic factors and insulin in preadipocytes, and ii) in the mechanisms underlying the variations in the proliferating and differentiating capacities of preadipocytes according to their anatomical localization.

Adipose Tissue↗

Enhancement of the expression of the alpha 2-adrenoreceptor protein and mRNA by a direct effect of androgens in white adipocytes.

In vivo, testosterone-treatment of female hamsters for 4 days promotes a doubling of alpha 2-adrenoreceptor protein in parametrial adipocytes, with a concomitant accumulation of the alpha 2A-adrenoreceptor subtype mRNA. During in vitro incubation of minced parametrial fat pads for 6 to 48h with testosterone or dihydrotestosterone (100 nM), alpha 2A-adrenoreceptor protein and mRNA levels were also increased and remained to control levels when an antiandrogen or actinomycin D were added in the medium. It is concluded that in hamster adipocytes, androgens upregulate alpha 2A-adrenoreceptor subtype expression at the mRNA level by an androgen receptor-dependent transcriptional activation.

Adipocytes↗

Estrogen binding sites in hamster white adipose tissue: sex- and site-related variations; modulation by testosterone.

Estrogen binding sites (ER) were studied, using 17 beta-[3H]estradiol as the ligand, in epididymal or parametrial and subcutaneous adipose tissues of male and female hamsters. Compared with other mammalian fat deposits, intact male and female hamsters possess abundant estrogen binding sites with moderate affinity for estradiol and which occur as a single class of receptor in males but as two populations in females. The levels of estrogen receptors depend on both sex and tissue localization. In males, receptor densities are higher in both localizations when compared to those of females and ER are more abundant in superficial adipose deposits than in the deep fat tissue. In females, there are two estrogen binding populations; the one with the highest affinity is similar to the classical estrogen receptor and both populations are more abundant in deep fat than in subcutaneous deposits, in contrast to male hamsters. These characteristics depend on androgen status: in male adipose tissues, testosterone (TP) up-regulates the ER levels. Conversely, in female fat deposits, TP down-regulates the highest affinity estrogen receptors and the lowest affinity population disappears. Binding affinities are never affected by testosterone. These results suggest that, in hamster adipose tissue, estrogen receptors exhibit site- and sex-related differences, as previously described for androgen receptors. Furthermore, estrogen receptor expression is modulated by the androgen status, depending on gender, which could be related to some physiological situations observed in the hamster.

Adipose Tissue↗

Differential sodium regulation between salt-sensitive and salt-resistant Sabra rats is not due to any mutation in the renal alpha 2B-adrenoceptor gene.

A defect in sodium modulation of density and agonist affinity of renal alpha 2-adrenoceptor exists in normotensive salt-resistant Sabra (SBN) rats when compared to hypertensive salt-sensitive (SBH). A highly conserved aspartic acid residue in the second helix has been implicated in sodium regulation of alpha 2-adrenoceptor-ligand interactions. As the alpha 2B-adrenoceptor subtype is preponderantly present in kidney of SBH and SBN rats, a mutation might distinguish this subtype between SBH and SBN rats. From this study, no difference between SBH and SBN alpha 2B-adrenoceptor gene could be demonstrated in terms of nucleotide sequence. These data suggest that in Sabra rats, the differential sodium regulation in density and agonist affinity between renal SBH and SBN alpha 2-adrenoceptor may have another origin than the alpha 2B-adrenoceptor encoding gene.

Amino Acid Sequence↗