Biomedical subjects

W Dong

Publications and source records attributed to W Dong.

At least 73 records · Page 4Linked to original sources

PubMed1998

Toxic metals stimulate inflammatory cytokines in hepatocytes through oxidative stress mechanisms.

Hepatocytes, as well as nonparenchymal cells, secrete proinflammatory cytokines and chemokines that are involved in the pathology of many liver diseases. In particular, tumor necrosis factor-alpha (TNFalpha), as well as members of the CXC family of chemokines, including interleukin (IL)-8 in humans and macrophage inflammatory protein (MIP)-2 in rodents, have been implicated in both damage and repair processes associated with various hepatotoxins. In the liver, cytokine secretion is usually associated with nonparenchymal cells, particularly Kupffer cells. In the present studies, cytokine gene expression and secretion were investigated in hepatocytes treated with cadmium chloride (CdCl2) or vanadium pentoxide (V2O5). Using human Hep G2 cells and freshly isolated rodent hepatocytes, it was demonstrated that metals increase gene expression and secretion of CXC chemokines and TNFalpha. IL-8 and MIP-2 secretion induced either by the metals or H2O2 were inhibited by antioxidants such as tetramethyl-thiourea and N-acetyl-cysteine. In vitro neutralization experiments with TNFalpha and in vivo studies with TNFalpha receptor knockout mice indicated that the metals directly stimulate CXC chemokine secretion without the need for TNFalpha. Taken together these studies indicate that, in addition to other inflammatory mediators and acute phase proteins, cytokines and chemokines are produced by hepatocytes, which may participate in hepatotoxic responses. The events responsible for their expression involve cellular redox changes.

Animals↗

PubMed1998

Enhanced allergic responses to house dust mite by oral exposure to carbaryl in rats.

Epidemiological studies have demonstrated an association between use of carbamate insecticides, including carbaryl, and increased incidence of allergic asthma in farmers. In this study the effect of oral carbaryl exposure on the development of allergic responses to house dust mite (HDM) was examined in female Brown Norway rats. Rats were gavaged for 2 weeks with 0, 2, 10, or 50 mg/kg/day of carbaryl. They were sensitized with a subcutaneous injection of HDM in aluminum hydroxide adjuvant 3 days after the beginning of carbaryl exposure and challenged with antigen via the trachea 1 day after the final carbaryl ingestion. Two days after challenge, antigen-specific cell proliferation in pulmonary lymph nodes was significantly higher in the 50 mg/kg group than in controls, while antigen-specific splenocyte proliferation was decreased in groups dosed with 2, 10, and 50 mg/kg carbaryl. Total protein and lymphocyte number in bronchoalveolar lavage (BAL) fluid were also increased in the 50 mg/kg group. By 7 days after challenge, immune-mediated pulmonary inflammation (eosinophils), antigen-specific immunoglobulin (Ig) E level in serum, and antigen-specific IgE and IgA levels in BAL fluid were significantly elevated in the 50 mg/kg group. No apparent change was observed for lactate dehydrogenase and eosinophil peroxidase in BAL fluid, while the number of BAL macrophages were decreased in groups dosed with 10 and 50 mg/kg carbaryl. The results suggest that carbaryl may cause systemic immune suppression, while enhancing pulmonary allergic responses to house dust mite antigen.

Animals↗

PubMed1998

Gender differences in accessing cardiac surgery across England: a cross-sectional analysis of the health survey for England.

OBJECTIVE: To examine gender differences in access to cardiac surgery in a nationally representative sample. DESIGN: Nationwide cross sectional household based survey (Health Survey for England). SETTING: Private households in England around 1993 and 1994. SUBJECTS: 1708 subjects reporting a history of either doctor diagnosed angina or heart attack from a stratified random sample of 32 378 people aged 16 and above. OUTCOME MEASURE: The proportion reporting having had cardiac surgery or on a waiting list. RESULTS: 13.5% reported previous (n = 206) or pending (n = 25) cardiac surgery. Men were more likely than women to have had or to be waiting for cardiac surgery (19.1% of men versus 6.8% of women, chi2 54.7, P<0.001). This finding was consistent regardless of age group and across three regional areas. The unadjusted odds ratio for cardiac surgery for men versus women was 3.3 (95% Cl 2.3, 4.5, P<0.001) and was only slightly attenuated to 2.8 (95% CI 1.9, 4.0. P<0.001), after adjustment for other factors. The gender difference remained even when analysis was restricted to subjects reporting a previous heart attack, and after statistical adjustment for disease severity. CONCLUSION: Women are less likely than men to receive cardiac surgery across all age groups and regional areas. These results include private operations and adjust for individual behavioural data. Neither disease severity or co-morbidity explains these discrepancies. Further studies are required to determine why this inequality occurs and how it can be addressed.

Adolescent↗

PubMed1998

Neuroendocrine protein 7B2 is essential for proteolytic conversion and activation of proprotein convertase 2 in vivo.

The 7B2 protein is widely distributed in neural and endocrine tissues. Its biological function was found to be related to the processing enzyme proprotein convertase 2 (PC2), a mammalian subtilisin/kexin-like endoproteinase that cleaves at specific single or multiple basic amino-acid residues. In order to examine the proposed function of 7B2 on PC2 in in vivo models, we first compared the distribution of 7B2 and PC2 mRNAs in the rat brain. Expression of 7B2 mRNA was found to be pan-neuronal, but additionally, we observed 7B2 mRNA in ependymal cells and in the subcommissural organ. Although the expression of PC2 mRNA was exclusively neuronal, it was more restricted, sparing some regions expressing high levels of 7B2. This finding suggests that 7B2 has an additional function in non-PC2-expressing cells. No evidence of PC2-positive/7B2-negative cells could be obtained in the adult rat brain. However, in the developing rat brain (E17), such regions were easily observed, showing higher levels of pro-PC2 (75 kD). Similarly, in the animal model of insulin-induced hypoglycemic shock, where adrenomedullary 7B2 expression is decreased, the ratio of pro-PC2 to mature PC2 (75 kD:68 kD) was observed to be increased. Finally, the human neuroepithelioma SK-N-MCIXC expresses PC2 but not 7B2. Accordingly, only inactive pro-PC2 forms were observed: 75-kD intracellular and 71-kD extracellular. After stable transfection of SK-N-MCIXC cells with 27-kD pro-7B2, mature and active (68-kD) PC2 was secreted into the medium. Our data demonstrate a critical role of 7B2 in the proteolytic conversion and activation of PC2 in vivo.

Adrenal Glands↗

PubMed1998

Cloning, sequence analysis, and distribution of rat metallocarboxypeptidase Z.

A cDNA encoding human carboxypeptidase Z (CPZ), a novel metallocarboxypeptidase, was recently cloned (Song and Fricker, J. Biol. Chem., 272, 1054, 1997). In the present study, a cDNA encoding the rat homolog of CPZ was identified. As with the human form, rat CPZ contains an N-terminal domain of 120 amino acids that has 20% to 30% amino acid identity with the "frizzled" domain found on proteins that interact with Wnt, a protein involved in tissue polarity in early embryogenesis. Sequence analysis showed rat and human CPZ to be highly conserved within the frizzled domain (77% amino acid identity), the carboxypeptidase domain (91%), and the C-terminal 28 residues (78%). The entire rat CPZ protein has high sequence similarity with human CPZ (81% amino acid identity), moderate sequence similarity to human carboxypeptidase N (45%), human carboxypeptidase E (41%), and human carboxypeptidase M (33%), and less sequence similarity with other metallocarboxypeptidases. Northern blot analysis showed rat CPZ mRNA to be abundant in the placenta, with low to moderate levels in the brain, lung, thymus, and kidney. The BRL3A rat liver cell line and the PC12 rat adrenal cell line express high levels of CPZ mRNA. In situ hybridization analysis indicated that CPZ is expressed only in specific cell types. For example, in the brain, CPZ mRNA is present in leptomeningeal cells, but not in the majority of other cell types. This distribution in leptomeningeal cells is shared by AEBP1, a recently reported member of the metallocarboxypeptidase gene family. However, the distribution of CPZ and AEBP1 differ in pituitary and thyroid. Taken together, these studies suggest that CPZ functions in a range of cell types.

Amino Acid Sequence↗

PubMed1998

Identification of mouse CPX-2, a novel member of the metallocarboxypeptidase gene family: cDNA cloning, mRNA distribution, and protein expression and characterization.

A novel member of the metallocarboxypeptidase gene family was identified from its homology with carboxypeptidase E and has been designated CPX-2. The cDNA of 2500 nucleotides encodes a protein of 764 amino acids that contains an N-terminal signal peptide-like sequence, a 158-residue discoidin domain, and a 400-residue carboxypeptidase domain. The 400-residue metallocarboxypeptidase domain has 59% amino acid identity with a protein designated AEBP-1; 44% to 46% identity with carboxypeptidases E, N, and Z; and lower homology with other members of the metallocarboxypeptidase gene family. The discoidin domain of CPX-2 has 22% amino acid identity with the carbohydrate-binding domain of discoideum-I, 29% to 34% identity with the phospholipid-binding domain of human factors V and VIII, and 59% identity with the discoidin-like domain on AEBP-1. CPX-2 is missing several of the predicted active-site residues that are conserved in most other members of the metallocarboxypeptidase gene family and which are thought to be required for enzyme activity. Expression of CPX-2 using the baculovirus system produced several forms of protein, from 80 to 105 kDa, but no detectable activity toward a variety of carboxypeptidase substrates. A shorter 50-kDa form of CPX-2, which contains the carboxypeptidase domain but not the discoidin domain, was also inactive when expressed in the baculovirus system. CPX-2 is able to bind to Sepharose-Arg; this binding is blocked by 10 mM Arg. Northern blot analysis showed CPX-2 mRNA in mouse brain, liver, kidney, and lung. In situ hybridization analysis of brain revealed a broad distribution. Areas that are enriched in CPX-2 include the hippocampus, cerebral cortex, median eminence, and choroid plexus. Taken together, these data suggest a widespread function for CPX-2, possibly as a binding protein rather than an active carboxypeptidase.

Amino Acid Sequence↗

PubMed1998

Cytokine expression in hepatocytes: role of oxidant stress.

Inflammatory mediators, including cytokines and chemokines, are associated with the pathology of chronic liver disease. Interleukin-8 (IL-8) in humans and macrophage inflammatory protein-2 (MIP-2) in rodents, both members of the C-X-C family of chemokines, are particularly potent neutrophil attractants and have been implicated in chronic liver diseases. In the liver, cytokine secretion is usually associated with non-parenchymal cells, particularly Kupffer cells. In the present studies, chemokine gene expression and secretion were investigated in hepatocytes treated with various stimulators. Using human Hep G2 cells, it was demonstrated that, in contrast to lipopolysaccharides (LPS), both tumor necrosis factor-alpha (TNF-beta) and H2O2 are potent inducers of IL-8, presumably acting via protein kinase C (PKC)-dependent pathways. MIP-2 expression occurred in freshly isolated rat hepatocytes following treatment with TNF-alpha, LPS, and to a lesser degree, H2O2. Both IL-8 and MIP-2 secretion were inhibited, although to varying degrees, by such antioxidants as TMTU, DMSO, catalase, and N-acetylcysteine. Furthermore, in vitro TNF-alpha neutralization experiments and transfection of Hep G2 cells with an IL-8 construct confirmed that TNF-alpha and H2O2 directly stimulate IL-8 secretion. RT-PCR analyses indicated that chemokine secretion induced by these agents operates via increased gene expression. Furthermore, a variety of cytokine genes were found to be expressed by hepatocytes, including MCP-1, cytokine-induced neutrophil chemoattractant (CINC), and IL-6. Taken together, these studies indicate that hepatocytes respond to biologically relevant levels of common activators, including H2O2, to produce cytokines and chemokines that contribute to pathophysiologic and repair processes in the liver.

Animals↗

PubMed1998

Blood pressure screening, management and control in England: results from the health survey for England 1994.

OBJECTIVE: To assess the current levels of awareness, treatment and control of hypertension in England and to determine the number and type of drugs prescribed. DESIGN: A cross-sectional household-based survey of English adults. SUBJECTS: A random sample from the adult English population of 12,116 adults who participated in the 1994 Health Survey for England. MAIN OUTCOME MEASURES: Prevalences of treatment hypertension, awareness and control. RESULTS: Using a definition of hypertension as a systolic blood pressure > or = 160 mmHg or a diastolic blood pressure > or = 95 mmHg, or a patient's being administered antihypertensive treatment, the prevalence of awareness of hypertension was 63%. Among hypertensives, 50% were receiving treatment and 30% had their hypertension controlled (< 160 mmHg/95 mmHg). Awareness, treatment and control rates are considerably lower than the most recently reported rates from the USA. Diuretics and beta-blockers remain the most common antihypertensive agents used in England. CONCLUSION: There is considerable scope for improving the treatment and control of hypertension in the English adult population.

Adolescent↗

PubMed1998

A virus-encoded RNA polymerase purified from baculovirus-infected cells.

A DNA-dependent RNA polymerase was purified to homogeneity, starting from insect cells infected with the baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV). The purified polymerase supported accurate and specific transcription from late and very late promoters but was not active on viral early promoters. Thus, promoter recognition is an integral function of the purified enzyme. The purified RNA polymerase was composed of only four equimolar subunits, which makes it the simplest DNA-directed RNA polymerase from a eukaryotic source described so far. Amino-terminal protein sequencing, peptide fingerprinting, and immunochemical analyses were used to identify the four subunits, all of which are virus encoded. Overexpression of the four viral proteins (LEF-8, LEF-4, LEF-9, and p47) in baculovirus-infected cells resulted in a significant increase in the levels of RNA polymerase produced in the infected cells. Thus, the overexpression data are consistent with our identification of the RNA polymerase subunits.

Amino Acid Sequence↗

PubMed1998

Guanylyltransferase activity of the LEF-4 subunit of baculovirus RNA polymerase.

The baculovirus Autographa californica nuclear polyhedrosis virus encodes a DNA-dependent RNA polymerase that transcribes viral late genes. This polymerase is composed of four equimolar subunits, LEF-4, LEF-8, LEF-9, and p47. Here we present data indicating that the LEF-4 subunit of RNA polymerase is a guanylyltransferase. Incubation of RNA polymerase in the presence of divalent cation and radiolabeled GTP resulted in the formation of a covalent enzyme-guanylate complex that comigrated with the LEF-4 subunit. The label transfer assay showed an absolute requirement for divalent cation which could be satisfied by either manganese or magnesium. The reaction was specific for guanine nucleotides, and GTP was more effective than dGTP in the formation of enzyme-guanylate complex. To demonstrate that LEF-4 was the guanylyltransferase, the single subunit was overexpressed in baculovirus-infected cells. The overexpressed protein was primarily cytosolic, indicating that other proteins in the RNA polymerase complex were responsible for nuclear targeting of LEF-4. LEF-4 alone was able to covalently bind GMP, although less efficiently than viral RNA polymerase.

Amino Acid Sequence↗

PubMed1998

The LEF-4 subunit of baculovirus RNA polymerase has RNA 5'-triphosphatase and ATPase activities.

The baculovirus Autographa californica nuclear polyhedrosis virus encodes a DNA-dependent RNA polymerase that is required for transcription of viral late genes. This polymerase is composed of four equimolar subunits, LEF-8, LEF-4, LEF-9, and p47. The LEF-4 subunit has guanylyltransferase activity, suggesting that baculoviruses may encode a full complement of capping enzymes. Here we show that LEF-4 is a bifunctional enzyme that hydrolyzes the gamma phosphates of triphosphate-terminated RNA and also hydrolyzes ATP and GTP to the respective diphosphate forms. Alanine substitution of five residues previously shown to be essential for vaccinia virus RNA triphosphatase activity inactivated the triphosphatase component of LEF-4 but not the guanylyltransferase domain. Conversely, mutation of the invariant lysine in the guanylyltransferase domain abolished the guanylyltransferase activity without affecting triphosphatase function. We also investigated the effects of substituting phenylalanine for leucine at position 105, a mutation that results in a virus that is temperature sensitive for late gene expression. We found that this mutation had no significant effect on the ATPase or guanylyltransferase activity of LEF-4 but resulted in a modest decrease in RNA triphosphatase activity.

Acid Anhydride Hydrolases↗

PubMed1998

Altered alveolar macrophage function in calorie-restricted rats.

Alveolar macrophage functions associated with clearance of bacteria from the lung were assessed in male Fischer 344 rats maintained on a 25% calorie-restricted diet. Calorie-restricted and ad libitum-fed (control) rats were exposed to concentrations of ozone known to compromise phagocytic function of alveolar macrophages. Ozone suppressed alveolar macrophage phagocytosis of latex beads in vitro in ad libitum-fed rats, but not in calorie-restricted rats. In fact, caloric restriction enhanced phagocytic function in both control and ozone-exposed animals. Ad libitum-fed rats exposed to ozone and challenged with Streptococcus zooepidemicus experienced a prolonged infection and influx of polymorphonuclear leukocytes (PMN), whereas calorie-restricted rats exposed to ozone cleared the bacteria in 24 h without an inflammatory response. Bacterial endotoxin-stimulated in vitro production of nitric oxide and tumor necrosis factor (TNF)-alpha as well as expression of TNF-alpha and interleukin-6 messenger RNAs were all lower in alveolar macrophages isolated from calorie-restricted rats. Together, the data suggest that caloric restriction enhances resistance to gram-positive bacteria, while lowering the production of proinflammatory mediators elicited by endotoxin, a component of gram-negative bacteria. Although increased bacterial resistance is considered beneficial, reduction in the lung's ability to induce inflammatory mediators can have both positive and pathophysiologic consequences.

Animals↗

PubMed1998

[Study on effect of Erigeron injection in prevention and treatment of cerebral ischemic injury].

OBJECTIVE: To study the effect of Erigeron injection (EI) on cerebral ischemic injury. METHODS: Animal model of brain ischemic/reperfusion was established in male Wistar rats and the effect of EI on proteinkinase C (PKC) activity, intraneuronal free calcium and regional cortex blood flow (rCBF) were observed. RESULTS: EI could prevent the activation of PKC, calcium overload and decrease of RCBF induced by cerebral ischemia/reperfusion. CONCLUSION: EI could prevent neuronal ischemic injury by inhibiting activation of PKC.

Animals↗

PubMed1998

[Experimental studies on antitumor activity of new recombinant human tumour necrosis factor in nude mice bearing nasopharyngeal carcinoma].

OBJECTIVE: Nude mice model bearing nasopharyngeal carcinoma were established using nasopharyngeal squamous carcinoma cell line (CNE-2). METHODS: New recombinant human tumor necrosis factor (NrhTNF-alpha) and carboplatin were administered locally or systemically, single or in combination. The microstructural and ultra-micro-structural changes of carcinoma after administration of NrhTNF-alpha were observed under light and electron microscopes. RESULTS AND CONCLUSION: 1. After treatment, the tumor showed hemorrhage, necrosis, regression, the survival time was prolonged. 2. Locally administered NrhTNF-alpha had more significant antitumor effect than intravenously administered of NrhTNF-alpha. 3. There is a notable synergistic antitumor effect by the combination use of NrhTNF-alpha and carboplatin. 4. Microscopy showed that cell changes mostly took place in plasma especially in mitochondria and endoplosmic reticula. The mitochondria were highly bloated. Besides, some nuclei were condensed and fragmented, resulted in cell death. These studies provide experimental basis for biological treatment of nasopharyngeal carcinoma.

Animals↗

PubMed1998

[Image and quantity analysis of prostaglandin in rats' blood plasma and Na(+)-K(+)-ATPase in their cerebellum during the prevention of motion sickness by cinnarizine].

To study the mechanism of cinnarizine in preventing motion sickness, TXB2, 6-Keto-PGF1 alpha in rats' blood plasma and Na(+)-K(+)-ATPase activity in the endothelial cells of their cerebellar capillary were measured and analysed by a radioactive immunity analyser and a computer image system. The results showed that TXB2 and 6-Keto-PGF1 alpha in rats' blood plasma in the cinnarizine preventing group (CPG) decreased remarkably, compared with those in the motion sickness group(MSG) (p < 0.05). The activity of Na(+)-K(+)-ATPase in the endothelial cells of rats' cerebellar capillary in CPG was higher than that in MSG (p < 0.01). The authors suggest that the lower concentration of TXB2 and 6-Keto-PGF1 alpha in rats' blood plasma in CPG is closely related to cinnarizine which prevents Ca2+ from entering into the platelets and into the endothelial cells of blood vessels. The higher activity of Na(+)-K(+)-ATPase in the cerebellum may be caused by cinnarizene which dilates the blood vessels in the brain, increases the blood flow therein, and hinders Ca2+ from getting into the cerebellum cells. These change are believed to be the important mechanism of how cinnarizine prevents motion sickness.

Animals↗

PubMed1997

Synthesis of Uniform Ferric Oxide Particles from Deionized Colloids

A modified method was employed to prepare monodispersed colloidal particles of ferric (hydrous) oxide. The method contains three steps: (i) preparation of uniform nuclei of ferric hydrous oxide via a so-called instantaneous nucleation method; (ii) purification of the nuclei suspension via dialysis; (iii) aging of the purified nuclei suspension in a reflux reactor at certain pH. Cubic and pseudocubic alpha-Fe2O3 monodisperse particles which are much smaller than the cubic alpha-Fe2O3 particles obtained from the same reactant through a usual method were produced by aging needle like nuclei at a lower pH. A close-packed three-dimensional QDs (quantum dots) superlattice structure 40 nm in edge length with cubic geometry was formed by self-aggregation between spherical amorphous ferric hydrous oxide nuclei 3-5 nm in diameter. The growth processes of the two kind particles were also illustrated. This study showed an approach to prepare smaller particles from aqueous metal salt solutions in relatively higher concentration, and ordered QDs superlattice structure by controlled self-aggregation of QDs in hydrosol.

Journal Article↗

PubMed1997

Ecological analysis of collectivity of alcohol consumption in England: importance of average drinker.

OBJECTIVE: To assess whether the average consumption of alcohol is associated with the prevalence of heavy drinking, problem drinking, and abstention in England. DESIGN: Ecological analysis using data from a cross sectional household based survey of English adults. SUBJECTS: Random sample of 32,333 adults from the English population who participated in the 1993 and 1994 health surveys for England. MAIN OUTCOME MEASURES: Association, expressed as the correlation coefficient, between the regional mean and median alcohol consumption and the regional prevalence of heavy drinking, problem drinking, and abstention. RESULTS: Mean consumption of alcohol in light to moderate drinkers was strongly positively associated with the prevalence of heavy drinking (r = 0.75 in men and r = 0.62 in women for drinking more than 21 and 14 units per week respectively). A similar association was found between median consumption and prevalence of heavy drinking. Abstention was not significantly associated with mean consumption in drinkers (r = 0.08 for men and r = -0.29 for women). Both the median and mean consumption in drinkers were positively associated with the prevalence of problem drinking as defined by the CAGE questionnaire on alcohol use (r = 0.53 for men and r = 0.42 for women for the association with mean consumption). CONCLUSION: Factors that increase the average consumption of alcohol in the population may result in an increase in the prevalence of heavy drinking and related problems.

Adult↗

PubMed1997

TGFbeta1 regulates gene expression of its own converting enzyme furin.

TGFbeta1 is known for its potent and diverse biological effects, including immune regulation, and cell growth and differentiation. We have recently shown that TGFbeta1 precursor is processed by human furin COOH-terminal to the R-H-R-R278 cleavage site to generate authentic mature TGFbeta1. In the present study, we demonstrate that steady-state furin mRNA levels are increased in rat synovial cells by 2 and 20 ng/ml TGFbeta1. Stimulation with TGFbeta1 results in a significant increase in furin mRNA levels, starting at 3 h with the peak effect observed at 12 h (2.5-fold increase +/-0.4). TGFbeta1 did not increase furin mRNA stability, and treatment of synovial cells with actinomycin D, before TGFbeta1 addition prevented the increase in fur gene expression, suggesting that the observed regulation occurs at the level of gene transcription. Treatment of synovial and NRK-49F fibroblastic cells with exogenous TGFbeta1 (5 ng/ml) or TGFbeta2 (10 ng/ml) translates into an increase in pro-TGFbeta1 processing as evidenced by the appearance of a 40-kD immunoreactive band corresponding to the TGFbeta1 NH2-terminal pro-region. Furin processing activity stimulated by TGFbeta2 correlates with significant increase in extracellular mature and heat-activable TGFbeta1 as determined by an isoform-specific ELISA assay. Taken together, these results demonstrate for the first time that TGFbeta1 upregulates gene expression of its own converting enzyme, and that this expression is translated into augmented processing of the TGFbeta1 precursor form. Such adaptive responsiveness of the TGFbeta1 convertase may represent an important aspect of TGFbeta1 bioavailibility in TGFbeta1-related processes and pathological conditions.

Amino Acid Sequence↗