Biomedical subjects

S Zhang

Publications and source records attributed to S Zhang.

At least 487 records · Page 27Linked to original sources

PubMed1999

Evaluation of a lumped parameter model for isolated working rat hearts.

When a mechanical model of the rat aortic input impedance is perfused with a pulsatile pump, the computed values of the impedance components vary linearly with flow rate and are interactive. When the model is perfused by an isolated rat heart, the total load upon the left ventricle consists of the model and coronary impedances in parallel. Adenosine triphosphate induces changes in coronary impedance, and the redistribution of cardiac output from the model to the coronary circulation causes flow-related changes in the model impedance. Thus, the mechanical model does not provide a constant load for the isolated heart, because of variations in both the model and coronary impedances.

Adenosine Triphosphate↗

PubMed1999

Effect of interleukin-1 beta on the elevation of cytoplasmic free calcium of the cultured hippocampal neurons induced by L-glutamate.

To investigate the intracellular mechanism that interleukin-1 beta (IL-1 beta) facilitates epileptic seizure and neuronal damage, the effect of IL-1 beta alone or IL-1 beta plus glutamate (Glu) on the intracellular free calcium ([Ca2+]i) of single cultured hippocampal neuron was examined by using EPC-9 light-electricity measurement system. The results showed that IL-1 beta of different concentrations (5 x 10(3) U/L, 10 x 10(3) U/L, 20 x 10(3) U/L, 30 x 10(3) U/L, 50 x 10(3) U/L, 100 x 10(3) U/L) failed to affect the neuronal [Ca2+]i, but IL-1 beta could facilitate the augmentation of neuronal [Ca2+]i induced by Glu in a dose-dependent pattern. MK-801 inhibited the effect of Glu on [Ca2+]i, and also inhibited the effect of IL-1 beta on [Ca2+]i induced by Glu, while verapamil did not influence the effect of Glu or IL-1 beta. It is concluded that IL-1 beta, as a neuromodulator, can facilitate the activation of NMDA receptor by Glu, induce the increase of intracellular calcium, which enhances the excitement of neuron.

Animals↗

PubMed1999

The Arabidopsis transposon Tag1 is active in rice, undergoing germinal transposition and restricted, late somatic excision.

Tag1 is an autonomous transposable element of Arabidopsis thaliana that displays tight developmental control of its excision during shoot development. To determine how Tag1 behaves in a monocotyledonous species, Tag1 was inserted in a 35S-GUS marker gene and the construct was introduced into rice. Tag1 showed somatic excision activity in four out of eleven transgenic lines examined. In leaves, excision was primarily restricted to vascular bundles and produced sectors composed of only a few cells. Excision events in flowers occurred predominantly in or near the major veins of the palea and lemma to produce small sectors. In roots, small sectors were evident, but they were few in number. These data show that the timing of Tag1 excision during rice shoot development is late and mimics the late excision behavior of Tag1 in Arabidopsis. One of the transgenic rice lines, which had a high frequency of somatic excision, produced several germinal revertants, one of which was characterized by a new Tag1 insertion band. The pattern of Tag1 transcripts and the footprint sequences left behind after excision in rice were found to be very similar to those in Arabidopsis. These results show that key properties of Tag1 transposition and behavior are conserved between monocots and dicots and that Tag1 has the potential to serve as an insertional mutagen in rice.

Arabidopsis↗

PubMed1999

Primary stenting in acute myocardial infarction: influence of diabetes mellitus in angiographic results and clinical outcome.

BACKGROUND: The outcome of patients with diabetes after myocardial infarction (MI) has traditionally been worse than in their nondiabetic counterparts before and during the thrombolytic therapy era. Whether the fate of patients with diabetes might improve with mechanical intervention, particularly with primary stenting, has not previously been studied. METHODS: We compared the angiographic and clinical outcome of 76 nondiabetic patients (aged 61 +/- 14 years; 66% male) and 28 patients with diabetes (aged 65 +/- 12 years; 64% male) consecutively treated with primary stenting for acute MI. Coronary Thrombolysis In Myocardial Infarction grade 3 flow was restored in 96% of diabetic and 97% of nondiabetic patients. RESULTS: Angiographic results after stent deployment were similar in the 2 groups. At 1-month follow-up, all patients in both groups were alive. Patients with diabetes had a much higher incidence of stent thrombosis (18% vs 1%; P =.003), which accounted for the majority of the major cardiac events at 1 month (21% vs 4%; P =.009). At a mean follow-up of 315 +/- 13 days, 99% of nondiabetic and 89% of patients with diabetes were alive (P =.04). Overall freedom from a major cardiac event (death, MI, target vessel revascularization) at 315 +/- 13 day follow-up was 88% for nondiabetics and 54% for patients with diabetes (P =.0003). By multivariate analysis, diabetes mellitus was the most important predictor for development of 1-month (RR 9.89; 95% confidence interval, 1.6-30) and late major cardiovascular events (RR 8.39; 95% confidence interval, 2.93-24). CONCLUSIONS: Primary stenting in acute MI is highly effective in restoring immediate TIMI 3 coronary flow in nondiabetic patients and patients with diabetes. This procedure may improve benefit in terms of mortality rate to both groups, particularly in patients with diabetes, compared with previous reports with thrombolytic therapy. Nevertheless, stent thrombosis and major cardiovascular events at 1 month and late follow-up are more frequent in patients with diabetes.

Aged↗

PubMed1999

Human antibody responses to Brugia malayi antigens in brugian filariasis.

Human antibody responses to Brugia malayi antigens were studied with sera from a Brugia endemic area in South India. Patients with clinical filariasis had significantly higher IgE and lower IgG4 levels to adult worm antigens than people with asymptomatic microfilaraemia. Intermediate antibody levels were observed in endemic normals. A majority of sera from each clinical group contained IgG antibodies to surface antigens of infective larvae (L3) by IFAT. IgG immunoblot studies did not reveal group differences in L3 antigen recognition. IgE antibodies bound to a subset of antigens bound by IgG. IgE antibodies in sera from clinical filariasis patients preferentially bound to L3 antigens at 200, 97, 68 and 58 kDa compared with sera from microfilaria carriers. These results are consistent with prior studies of antibody responses in filariasis and add new information on the targets of IgG and IgE antibodies to L3 antigens in brugian filariasis.

Adult↗

PubMed1999

Potentiation of recombinant L-type Ca channel currents by alpha1-adrenoceptors coexpressed in baby hamster kidney (BHK) cells.

In cardiac myocytes, the effect of alpha1-adrenergic stimulation on L-type Ca current remains to be clarified. We examined this issue by the transient coexpression of alpha1-adrenoceptors on BHKC12 cells, where recombinant Ca channels composed of cardiac alpha1 subunit and skeletal beta, gamma, alpha2/delta subunits were stably expressed. After transfection of plasmid DNA encoding bovine alpha1C-adrenoceptors, bath-applied phenylephrine potentiated the cloned Ca channel current during perforated-patch whole-cell recording by 26+/-6% in 6 out of 12 cells. The potentiation was elicited also by methoxamine, and was blocked by prazosin. Phenylephrine also increased the channel open probability during cell-attached single channel recording in 7 out of 15 cells. The ratio of successful modulation of Ca channels was in accordance with the ratio of successful expression of alpha1-adrenoceptors, as estimated by beta-galactosidase staining. These results suggest that the stimulation of alpha1C-adrenoceptors is linked to potentiation of cardiac L-type Ca current. BHK cells provide a valuable expression system to study the modulation of Ca channels evoked by a receptor stimulation.

Animals↗

PubMed1999

Titanium dioxide as photocatalyst on porous nickel: adsorption and the photocatalytic degradation of sulfosalicylic acid

The commonly used photocatalyst, TiO2 (anatase), has been immobilized on porous nickel using 3 wt.% polyvinyl alcohol (PVA) as the binder. The results show that sulfosalicylic acid (SSal) can be degraded on the developed catalytic system. The adsorption characteristics on TiO2-Ni system have been investigated. The observance of photocalytic degradation of SSal under pH values and initial concentrations can be explained by the adsorption behavior of SSal. The parameters of the Langmuir-Hinshelwood expression have been determined by different experimental ways and the results are satisfactory.

Journal Article↗

PubMed1999

Continuous monitoring of brain tissue oxygen pressure in patients with severe head injury during moderate hypothermia.

BACKGROUND: We studied the change of brain tissue oxygen pressure (PbrO2) value within 24 hours after trauma and during moderate hypothermia in patients with severe head injury. The PbrO2 value was used to differentiate patients at risk of brain ischemia and to predict outcome. METHODS: A flexible microcatheter to be used for continuous monitoring of brain tissue oxygen was inserted into the normal frontal white matter, along with a thermocouple, in 14 patients with severe head injury within 1.5-12 hours (mean 6.1 +/- 5.2 hours) after trauma. Moderate hypothermia was also used within 24 hours in these patients. RESULTS: (1) No complications related to the microcatheter were seen. (2) Low initial PbrO2 values (mean values <10 mmHg) were noted after severe head injury. (3) The occurrence of low initial PbrO2 values (< or = 5 mmHg) was significantly correlated with a poor outcome. (4) Moderate hypothermia can increase PbrO2, but hyperventilation reduced PbrO2. (5) The difference between RT and BT increased during moderate hypothermia. CONCLUSIONS: The PbrO2 values were low in severe head injury, but hypothermia may improve these values. The technique of continuously monitoring brain tissue oxygen pressure may give better insight into cerebral oxygenation and warn of impending ischemia of brain tissue, especially in patients treated with hyperventilation. It will help to improve the management and final outcome of patients with severe head injuries.

Adult↗

PubMed1999

Biological surface engineering: a simple system for cell pattern formation.

Biological surface engineering using synthetic biological materials has a great potential for advances in our understanding of complex biological phenomena. We developed a simple system to engineer biologically relevant surfaces using a combination of self-assembling oligopeptide monolayers and microcontact printing (muCP). We designed and synthesized two oligopeptides containing a cell adhesion motif (RADS)n (n = 2 and 3) at the N-terminus, followed by an oligo(alanine) linker and a cysteine residue at the C-terminus. The thiol group of cysteine allows the oligopeptides to attach covalently onto a gold-coated surface to form monolayers. We then microfabricated a variety of surface patterns using the cell adhesion peptides in combination with hexa-ethylene glycol thiolate which resist non-specific adsorption of proteins and cells. The resulting patterns consist of areas either supporting or inhibiting cell adhesion, thus they are capable of aligning cells in a well-defined manner, leading to specific cell array and pattern formations.

3T3 Cells↗

PubMed1999

Smokeless tobacco and nicotine bring about excessive cytokine responses of murine memory T-cells.

To determine the influence of smokeless tobacco (ST) and nicotine on the cytokine phenotype of memory T-cells, splenic mononuclear cells (SPM) were exposed to 1:10(2) or 1:10(3) dilutions of ST extract (ST-SPM), 10 or 100 microg/ml nicotine (NIC-SPM), or medium (CON-SPM) during 4 days of stimulation with anti-CD3. SPM were then washed extensively to remove residual ST or nicotine and restimulated with anti-CD3 and anti-CD28 in the absence of ST or nicotine. Expression of IL-2, IL-4, IL-10 and IFN-gamma protein and mRNA levels after 4 days of primary stimulation and after 24 and 48 h of restimulation was evaluated using ELISA and RT-PCR, respectively. After 4 days of primary stimulation, SPM exposed to 100 microg/ml nicotine sustained expression of IL-2, IFN-gamma, IL-10 and IL-4 mRNA as opposed to CON-SPM. Restimulation of CON-SPM resulted in maximum re-expression of cytokine mRNA at 24 h and a decline by 48 h. Restimulated NIC-SPM in the absence of nicotine delayed maximal re-expression of IL-2, IFN-gamma, IL-10 and IL-4 mRNA until 48 h. Heightened expression of cytokine mRNA at 48 h was paralleled by a small but significant increase in production of IFN-gamma, IL-4 and IL-10 protein by NIC-SPM as measured by ELISA. In contrast, ST-SPM did not exhibit residual expression of cytokine mRNA after 4 days of primary stimulation. Like NIC-SPM, however, restimulated ST-SPM exhibited maximum IL-2, IL-4, IFN-gamma, and IL-10 mRNA at 48 h. Heightened re-expression of cytokine mRNA at 48 h by ST-SPM was paralleled by increased production of IL-2, IFN-gamma, IL-4 and IL-10 protein. These results indicate that exposure of T-cells to nicotine, but not ST, during a primary immune response result in inordinate cytokine expression after 4 days. In addition, memory T-cells initially exposed to nicotine or ST during a primary immune response, exhibit excessive cytokine expression when T-cells are restimulated in the absence of nicotine or ST. pharmacology.

Animals↗

PubMed1999

Electrically-evoked inhibitory effects of the nucleus submedius on the jaw-opening reflex are mediated by ventrolateral orbital cortex and periaqueductal gray matter in the rat.

In previous studies we have shown that electrical stimulation of the nucleus submedius inhibits the rat radiant heat-induced tail flick reflex, and that this antinociceptive effect is mediated by the ventrolateral orbital cortex and periaqueductal gray. The aim of the present study was to examine whether electrical stimulation of the nucleus submedius could inhibit the rat jaw-opening reflex, and to determine whether electrolytic lesions of the ventrolateral orbital cortex or the periaqueductal gray could attenuate the nucleus submedius-evoked inhibition. Experiments were performed on pentobarbital-anesthetized rats. The jaw-opening reflex elicited by electrical stimulation of the tooth pulp or the facial skin was monitored by recording the evoked digastric electromyogram. Conditioning stimulation was delivered unilaterally to the nucleus submedius 90 ms prior to each test stimulus to the tooth pulp. After that, electrolytic lesions were made in ventrolateral orbital cortex or periaqueductal gray, and the effect of nucleus submedius stimulation on the jaw-opening reflex was re-examined. Unilateral electrical stimulation of nucleus submedius was found to significantly depress the jaw-opening reflex (mean threshold of 28.0+/-1.4 microA, n = 48), and the magnitude of inhibition increased linearly when the stimulus intensity was increased from 20 to 70 microA, resulting in depression of the digastric electromyogram amplitude from 18.4+/-5.4% to 74.0+/-4.9% of the control (P < 0.01, n = 37). The onset of inhibition occured 60 ms after the beginning of nucleus submedius stimulation and lasted about 100 ms, as determined by varying the conditioning-test time interval. Furthermore, ipsilateral lesions of the ventrolateral orbital cortex or bilateral lesions of the lateral or ventrolateral parts of periaqueductal gray eliminated the nucleus submedius-evoked inhibition of the jaw-opening reflex. These data suggest that the nucleus submedius plays an important role in modulation of orofacial nociception, and provide further support for a hypothesis that the antinociceptive effect of nucleus submedius stimulation is mediated by ventrolateral orbital cortex and activation of a descending inhibitory system in the periaqueductal gray.

Animals↗

PubMed1999

Protective role of nitric oxide in ischemia and reperfusion injury of the liver.

BACKGROUND: The suppressed production of nitric oxide (NO), associated with endothelial dysfunction, is thought to be a cause of ischemia and reperfusion injury of the liver. But findings of the salutary effects of NO enhancement on such injury have been conflicting. In this study, we tested our hypothesis that NO enhancement would attenuate ischemic liver injury. For this purpose, an NO precursor, L-arginine, and a novel NO donor, FK409, were applied to a 2-hour total hepatic vascular exclusion model in dogs. STUDY DESIGN: L-arginine was administered IV at a dose of 100 mg/kg twice (n = 5), while 300 mg/kg twice of FK409 was infused continuously into the portal vein (n = 5). The drugs were given to the animals for 30 and 60 minutes before and after ischemia, respectively. Non-treated animals were used as the control (n = 10). Two-week survival, systemic and hepatic hemodynamics indices, liver function tests, energy metabolism, and histopathology were analyzed. RESULTS: Both treatments comparably augmented hepatic tissue blood flow, decreased liver enzyme release, and increased high-energy phosphate restoration during the reperfusion period, all of which contributed to rescuing all of the treated animals from the 2-hour total hepatic ischemia. In contrast, ischemia caused 70% mortality in the control group. Histologically, structural abnormality and neutrophil infiltration were markedly attenuated by the treatments. Systemic hypotension was observed in the animals treated with FK409, however. CONCLUSIONS: Our data demonstrate that NO enhancement alleviates the liver injury caused by ischemia and reperfusion. The supplementation of L-arginine, rather than FK409, is considered more applicable to clinical use because of the absence of systemic adverse effects.

Adenine Nucleotides↗

PubMed1999

Is the Fischer 344/CRJ rat a protein-knock-out model for dipeptidyl peptidase IV-mediated lung metastasis of breast cancer?

Fischer 344/CRJ rats harbor a G633R substitution in dipeptidyl peptidase IV (DPP IV) that leads to retention and degradation of the mutant protein in the endoplasmic reticulum (Tsuji E, Misumi Y, Fujiwara T et al. Biochemistry 1992; 31 (47): 11921-7). However, when these rats were used as a 'protein knock-out' model in further evaluating the previously established role of DPP IV in metastasis, lung colonization of the highly metastatic MTF7 rat breast cancer cell line was reduced by only 33% relative to normal Fischer 344 rats. To examine whether lung endothelia leak expression of mutant DPP IV and whether mutant DPP IV exhibits the same adhesion qualities as wild type DPP IV, detailed immunohistochemical, biochemical, transfection, and FACS analyses were performed to assess the surface expression of mutant DPP IV on lung endothelia and transfected HEK293 cells and adhesion assay to compare the adhesion qualities of wild-type and mutant DPP IV. Both endothelial and transfected HEK293 cells expressed mutant, enzymatically inactive DPP IV on their surfaces, albeit at greatly reduced levels when compared to expression of wild type DPP IV. Purified mutant DPP IV had identical adhesion qualities for lung-metastatic MTF7 cells as wild type DPP IV, and competitive inhibition of MTF7 lung colonization by truncated DPP IV confirmed involvement of mutant DPP IV in lung metastasis of Fischer 344/CRJ rats. Although metastasis appears to be mediated by several, often parallel mechanisms involving multiple tumor and host factors, these data indicate that altered expression of a single component can drastically change the outcome of metastatic disease.

Adenocarcinoma↗

PubMed1999

Cognitive behavioral therapy of minor depressive symptoms in elderly Chinese Americans: a pilot study.

There is a high prevalence of suicide among elderly Chinese, and particularly among elderly Chinese women in Mainland China with a prevalence of 19.6 per hundred thousand. Since Chinese individuals may much more highly value education, a cognitive-behavioral package originated by Ricardo Munoz, Ph.D. was adapted for Chinese American subjects. The material was videotaped in eight sessions, approximately 25 minutes in length, to be shown to community subjects who were at least 40 years and over. In addition, a videotape of muscular relaxation techniques was made. A manual written in Chinese about the content of each class, was given to each subject when he/she attended. The experimental group showed significant improvement in the scores in the Hamilton Depression Scale, including the Somatic Subscale in the Hamilton Anxiety Scale. There was no significant improvement in the control group on any of the measures. Thus the study suggests the efficacy of psychoeducational classes in reducing symptoms of depression in non-patient community elderly. Other studies are being conducted among Korean Americans and Japanese Americans in the United States, and also in the Orient among Japanese elderly.

Aged↗

PubMed1999

HLA-DR-DQ alleles and HLA-DP alleles are independently associated with susceptibility to different stages of post-schistosomal hepatic fibrosis in the Chinese population.

Abstract: Evaluation of human leukocyte antigen (HLA) class I and class II genes was performed on patients from China with Schistosomiasis japonica. Patients were categorized as grade 0 (n=44), grade I (n=81), grade II (n=99), or grade III (n=6) based on increasing severity of hepatic fibrosis due to repeated Schistosoma japonicum infections. These results show that the HLA-DRB1*1101-DQA1*0501-DQB1*0301 (Pc<0.02) and HLA-DRB1*1501-DRB5*0101 (Pc<0.02) haplotypes are associated with protection and susceptibility to grade I fibrosis, respectively, and that the HLA-DPA1*0103 -DPB1*0201 haplotype (Pc<0.02) is associated with protection from both grade II and III severe fibrosis. There was no association between HLA-B DNA haplotypes and the disease. These findings indicate that the HLA-class II molecules play a role in preventing or promoting fibrotic liver change after deposition with Schistosome eggs. Moreover, a tendency was observed within the HLA class II genes for the HLA-DR-DQ alleles to be associated with protection against early changes in liver fibrosis, whereas HLA-DP alleles were associated with protection from the late phase of fibrosis or severe hepatosplenic schistosomiasis.

Alleles↗

PubMed1999

Human homologue of the Drosophila melanogaster lats tumour suppressor modulates CDC2 activity.

We have previously used mosaic flies to screen for tumour suppressors or negative regulators of cell proliferation. The cellular composition of these flies resembles that of cancer patients who are chimaeric individuals carrying a small number of mutated somatic cells. One of the genes we identified is the large tumour suppressor gene, lats (also known as wts), which encodes a putative serine/threonine kinase. Somatic cells mutant for lats undergo extensive proliferation and form large tumours in many tissues in mosaic adults. Homozygous mutants for various lats alleles display a range of developmental defects including embryonic lethality. Although many tumour suppressors have been identified in Drosophila melanogaster, it is not clear whether these fly genes are directly relevant to tumorigenesis in mammals. Here, we have isolated mammalian homologues of Drosophila lats. Human LATS1 suppresses tumour growth and rescues all developmental defects, including embryonic lethality in flies. In mammalian cells, LATS1 is phosphorylated in a cell-cycle-dependent manner and complexes with CDC2 in early mitosis. LATS1-associated CDC2 has no mitotic cyclin partner and no kinase activity for histone H1. Furthermore, lats mutant cells in Drosophila abnormally accumulate cyclin A. These biochemical observations indicate that LATS is a novel negative regulator of CDC2/cyclin A, a finding supported by genetic data in Drosophila demonstrating that lats specifically interacts with cdc2 and cyclin A.

Amino Acid Sequence↗

PubMed1999

Specificity analysis of sera from breast cancer patients vaccinated with MUC1-KLH plus QS-21.

The mucin MUC1 is expressed on breast cancers in an underglycosylated form compared to normal tissues and is therefore a potential target for cancer immunotherapy. MUC1 contains multiple tandem repeats of the 20 amino acid (aa) peptide (VTSAPDTRPAPGSTAPPAHG). The APDTRPA epitope is particularly immunogenic since it is recognized by a variety of murine monoclonal antibodies and by some sera and cytotoxic T-cells from unimmunized patients with epithelial cancers. We have prepared a 30 aa peptide (C)VTSAPDTRPAPGSTAPPAHGVTSAPDTRPA with cysteine at the N-terminal end, and used the cysteine for chemical conjugation to keyhole limpet haemocyanin (KLH). Six breast cancer patients immunized with this conjugate plus the immunological adjuvant QS-21 have all produced high titre (by ELISA) IgG and IgM antibodies against the 30 aa MUC1 peptide, but these sera reacted moderately, or not at all, with MUC1-positive tumour cells. To understand this specificity better, we prepared a series of smaller peptides to determine the epitopes recognized by these immune sera in inhibition assays. Only peptides containing APDTRPA at the C-terminal end were able to completely inhibit ELISA reactivity for the full 30 aa peptide. No sera were completely inhibited by APDTR, APDTRP, PDTRPA or any other peptides that did not contain the full APDTRPA epitope. Remarkably, sera from all six patients recognized this same epitope and were completely inhibited by only this epitope. The specificity of these sera (1) primarily for C-terminal APDTRPA, and the absence of this epitope at the C-terminal end of any tumour mucins, and (2) the N-terminal APDTRPA alanine, which is normally buried in the beta turn MUC1 assumes in its secondary structure may explain the moderate to weak reactivity of these high titer sera against MUC1-positive tumour cells.

Amino Acid Sequence↗

PubMed1999

Molecular cloning and expression of Galbeta1,3GalNAc alpha2, 3-sialyltransferase from human fetal liver.

Based on the sequences of the highly conserved segments in the previously cloned sialyltransferases, a cDNA encoding Galbeta1, 3GalNAc alpha2,3-sialyltransferase (SIATFL) has been isolated from human fetal liver. Expression analysis of the gene has been performed with various carcinoma cell lines, fetal tissues, fetal and adult liver and both hepatoma and the surrounding tissue from the same liver. The SIATFL gene was expressed poorly in fetal liver and in adult liver, slightly in hepatoma and highly in the surrounding tissue of hepatoma. The cDNA encoding the putative active domain was expressed in COS-1, Escherichia coli, and Pichia pastoris. The recombinant protein expressed in COS-1 could catalyse the transfer of NeuAc from CMP-NeuAc to asialo-fetuin. No enzyme activity was detected with a 32-kDa protein in E. coli and both 32-kDa and 41-kDa proteins in P. pastoris. These results suggested that correct glycosylation of the enzyme might play a key role in its folding that may be directly related to the enzymatic activity.

Amino Acid Sequence↗