Biomedical subjects

B Liu

Publications and source records attributed to B Liu.

At least 613 records · Page 34Linked to original sources

PubMed1994

hMSH2 mutations in hereditary nonpolyposis colorectal cancer kindreds.

It has recently been shown that hereditary nonpolyposis colorectal cancer (HNPCC) is caused by hereditable defects in DNA mismatch repair genes. However, the fraction of HNPCC due to defects in any one repair gene and the nature of these mutations are not known. We analyzed 29 HNPCC kindreds for mutations in the prototype DNA mismatch repair gene hMSH2 by a combination of linkage analysis, polymerase chain reaction-based screening, and sequencing of the coding region. The complete intron/exon structure of the gene was ascertained to facilitate this analysis. The results suggest that at least 40% of classic HNPCC kindreds are associated with germline mutations in hMSH2 and that most of these mutations produce drastic alterations in the predicted protein product.

Amino Acid Sequence↗

PubMed1994

12(S)-HETE enhancement of prostate tumor cell invasion: selective role of PKC alpha.

BACKGROUND: Prostate carcinoma has become the second most fatal cancer in American men. In Dunning R3327 rat prostate adenocarcinoma cells, elevated invasiveness positively correlates with metastatic potential. However, the mechanism(s) responsible for regulation of tumor cell motility and invasion is poorly understood. We have reported that a lipoxygenase metabolite of arachidonic acid, 12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE], augments tumor cell metastatic potential through activation of protein kinase C (PKC). PURPOSE: We proposed to determine the effect of 12(S)-HETE on the motility and invasion of low-metastatic rat prostate AT2.1 tumor cells and the effect of 12(S)-HETE activation of specific PKC isoform(s) in these processes. METHODS: The motility of AT2.1 cells was determined by the colloidal gold phagokinetic track assay and the invasiveness measured as their ability to invade through basement membrane Matrigel-coated filters. Expression of PKC isoforms was determined by Western blotting of the whole cell lysate with isoform-specific anti-PKC antibodies. Cytosol and membrane fractions were prepared and the subcellular distribution of PKC was analyzed by Western blotting and activity assay. The effect of 12(S)-HETE on cell proliferation was examined. Data were analyzed for significance of difference with the two-sampled, two-sided Student's t test. RESULTS: 12(S)-HETE increased the motility and invasion of AT2.1 cells, and this 12(S)-HETE-increased motility and invasion were inhibited by a selective PKC inhibitor, calphostin C, as well as a Ca2 chelator, bis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/tetra(acetoxy-methyl)ester. AT2.1 cells expressed the PKC isoforms alpha and delta, and 12(S)-HETE increased the membrane association of PKC alpha but not delta. Further, the motility and invasion of AT2.1 cells were increased by thymelea toxin, a selective activator of PKC alpha over PKC delta. CONCLUSION: 12(S)-HETE augments the invasiveness of AT2.1 cells via selective activation of PKC alpha. IMPLICATIONS: 12(S)-HETE modulation of PKC alpha invasiveness may be an important mechanism of action for the regulation of the invasive potential of rat prostate carcinoma cells, and the 12-lipoxygenase enzyme and/or PKC alpha may serve as key targets for the development of anti-invasive agents useful for combating the spread of prostate cancer.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗

PubMed1994

Platelet-derived growth factor A-chain gene transcription is mediated by positive and negative regulatory regions in the promoter.

Platelet-derived growth factor (PDGF) is a disulphide-linked heterodimer of two polypeptide chains, the A and B chains, which are encoded by genes on separate chromosomes. The A-chain gene is transcribed in a number of transformed and non-transformed cell lines and is inducible by a wide variety of growth factors, cytokines and other mitogenic agonists. To localize DNA elements that mediate basal transcription in the promoter regulatory region of the A-chain gene, we have employed 5'-endpoint deletion mutagenesis and transient expression analysis in the renal epithelial cell line BSC-1 (African green monkey). Studies conducted in this cell line, which expresses high concentrations of PDGF A-chain mRNA, reveal a positive regulatory element (PRE) in a GC-rich stretch of the A-chain promoter between -82 and -40, relative to the transcription start site. Two discrete regions of the promoter were identified as negative regulatory elements (NREs), located between -1029 and -880 (NRE1) and between -1800 and -1029 (NRE2). The -1800 to -812 region, which contains both NREs, functions as a potent NRE when relocated in either orientation adjacent to the herpes simplex virus thymidine kinase promoter, reducing transcription activity by 60% in the positive orientation and 85% in the negative orientation. Comparison of BSC-1 cells and Saos-2 cells (human osteogenic sarcoma), which do not express significant quantities of PDGF A-chain mRNA or protein, indicates that basal transcription of the gene is determined by enhancer activity mediated by the GC-rich region rather than through de-repression of the upstream NREs. Electrophoretic gel mobility shift assays reveal a complex pattern of nuclear protein binding to the GC-rich PRE (-73 to -46). Competition studies conducted with mutant oligonucleotides that alternately disrupt consensus binding sites for Sp-1 or Egr-1 demonstrate a requirement for the presence of an Sp1-like core sequence (GGCGGG) but not Egr-1/Krox-24 [GCG(G/T)-GGGCG] for the formation of specific DNA-protein complexes. Our observations suggest that basal transcription of the A-chain gene in renal epithelial cells is achieved through active enhancement, mediated by a GC-rich PRE and nuclear proteins that bind to Sp-1-like consensus DNA sequences.

Animals↗

PubMed1994

Functional morphological alterations of human blood platelets induced by oxidized low density lipoprotein.

The effect of oxidized low density lipoprotein (LDL) on the functional morphology of human platelets in vitro was studied by means of transmission electron microscopy. The washed platelets, stimulated by oxidized LDL (50-300 micrograms protein/ml), showed disc-sphere transformation, centralization of granules and complete degranulation in a dose- and time-dependent manner. A cytodamage in platelet membrane was induced by oxidized LDL leading to a lower electron density of cytoplasm compared to control. The morphological observations were supported by an analysis of the platelet shape-change parameter. Since the shape change, induced by oxidized LDL (50 micrograms/ml), was inhibited by a preincubation of platelets with staurosporine (10 nM), the protein kinase C was probably involved in the platelet activation initiated by oxidized LDL. The present results suggest that oxidized LDL could contribute to pathological thrombosis and atherogenesis by activating platelets.

Blood Platelets↗

PubMed1994

Endogenous 12(S)-HETE production by tumor cells and its role in metastasis.

12(S)-Hydroxyeicosatetraenoic acid [12(S)-HETE] is the 12-lipoxygenase metabolite of arachidonic acid. Previously, we have demonstrated that exogenous 12(S)-HETE can activate protein kinase C, increase cell surface expression of integrins, enhance adhesion, induce endothelial cell retraction, and increase experimental metastasis of tumor cells. Because of these prominent effects of exogenous 12(S)-HETE on tumor cell metastatic potential, it is important to determine whether there is endogenous 12(S)-HETE production by tumor cells. In the present study, mRNAs from human, rat, and mouse platelets as well as human colon carcinoma (Clone A), rat Walker carcinoma (W256), and mouse melanoma (B16a) and lung carcinoma (3LL) were reverse transcribed and amplified by polymerase chain reaction with platelet 12-lipoxygenase specific primers. Identity of the polymerase chain reaction fragments was confirmed by sequencing. 12-Lipoxygenase protein was detected by Western blotting. Tumor cell-derived 12-HETE was determined by reverse phase-high performance liquid chromatography analysis. In addition, the effect of endogenous 12(S)-HETE on tumor cells was studied by using a platelet-type 12-lipoxygenase selective inhibitor (N-benzyl-N-hydroxy-5-phenylpentanamide). Our results suggest that some tumor cells express platelet-type 12-lipoxygenase mRNA, protein and metabolize arachidonic acid to 12(S)-HETE and that endogenous 12(S)-HETE, like the exogenous 12(S)-HETE, may play an important role in tumor cell adhesion to matrix in vitro and lung colonization in vivo.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗

PubMed1994

Human amnion collagen for soft tissue augmentation--biochemical characterizations and animal observations.

As bovine collagen is currently being scrutinized as to its immunogenicity in clinical use, a human source collagen, human amnion collagen (HAC), has been developed in our laboratory as an injectable biomaterial for soft tissue augmentation. Pepsin-extracted human amnion collagen was highly purified and reconstituted. Gamma irradiation was employed to ensure complete sterility and to produce cross-linking in collagen chains to improve implant persistence without the use of chemical additives. The purity and characteristics of human amnion collagen were proven by amino acid assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immune blotting, and collagenase digestion. Animal studies comparing both irradiated and nonirradiated amnion collagen to bovine collagen (Zyderm and Zyplast) were carried out in a rat model. Humoral immunity was evaluated by examining the sera for antibody reactivity towards the implanted human collagen by the ELISA test. Insignificant antibody levels against human amnion collagen were found. Animal observation revealed fibroplasia, vascular infiltration, and the development of adipocytes with the implant as well as a lack of inflammatory response following up to 12 months of implantation. The persistence rate of our human amnion collagen was equal to, or even longer than, that of both types of bovine collagen implants.

Amino Acids↗

PubMed1994

Improved rat heart function after low Ca2+ perfused preservation at 5 degrees C for 18 hours.

To evaluate the effect of Ca2+ on recovery of heart function after hypothermic preservation, isolated working rat hearts were perfused continuously in different [Ca2+] (0.35-2.0 mM) at 37 degrees C for 30 min, during cooling to 5 degrees C, and then at 5 degrees C for 1, 6 or 18 h. After rewarming to 37 degrees C, contractile functions were assessed in working hearts perfused with Krebs-Henseleit (K-H) solution containing 2.0 mM free Ca2+. In normal K-H solution (2.0 mM Ca2+), cardiac function recovered completely after perfusion at 5 degrees C for 1 h (measured by the maximal rate of pressure development, +dP/dtmax, and heart rate x left ventricular systolic pressure, HR x LVSP). After 6 h at 5 degrees C, significant (P < 0.05) loss in function began to appear. Further, 18 h at 5 degrees C resulted in significant decreases in LVSP, +dP/dtmax, and coronary flow (CF) and a significant increase in left ventricular diastolic pressure. The deterioration of heart function consequent to hypothermic preservation could be prevented by reducing the Ca2+ in perfusion medium prior to and during cooling and rewarming from hypothermia. The optimal Ca2+ concentration was 0.5 mM. At lower Ca2+ (0.35 mM), reductions in HR, LVSP. +dP/dtmax, and CF after rewarming were evident. Our results indicate that by reducing cellular Ca2+ prior to cooling through perfusion of 0.5 mM Ca2+ K-H solution, significant benefits can be obtained both in prolonging the duration of cardiac preservation and in retaining functional integrity upon rewarming.

Animals↗

PubMed1994

Sex differences and effects of estrogen on dopamine and DOPAC release from the striatum of male and female CD-1 mice.

In the present study, we directly compare striatal dopamine metabolism in gonadectomized male and female CD-1 mice treated with 2 days of estrogen or oil vehicle. Basal and potassium-stimulated dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC) release from in vitro superfused striatum as well as pre- and postsuperfusion tissue dopamine contents were measured. Both basal and potassium-stimulated dopamine release were significantly higher and DOPAC release was significantly lower in males than in females. However, striatal tissue dopamine content was lower in males than in females. Estrogen-treated female mice showed increased basal and potassium-stimulated dopamine release compared to oil-treated females without affecting tissue dopamine content. Estrogen did not affect striatal dopamine concentrations or release in males. These results demonstrate clear sex differences in striatal dopamine turnover and concentrations under conditions of equal hormonal status. The results also indicate that estrogen can exert substantial effects on striatal dopamine metabolism by acting specifically in females to increase neuronal dopamine synthesis and release without depleting dopamine content. These results have important implications for the observed sex differences in clinical movement disorders such as Parkinson's disease.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1994

Effects of estrogen treatment on sensorimotor task performance and brain dopamine concentrations in gonadectomized male and female CD-1 mice.

In Experiment I, castrated male and female CD-1 mice +/- estradiol benzoate (EB) treatment were tested for their performance on a skilled sensorimotor task consisting of walking across beams of varying widths (6, 9, 12, and 21 mm). To evaluate whether behavioral performance was related to nigrostriatal dopaminergic function, tissue dopamine concentrations were determined from the corpus striatum as well as the hypothalamus and olfactory tubercle. In general, sensorimotor performance improved for all treatment conditions as the beam width increased. Castrated male mice treated with oil vehicle showed the worst performance as indicated by significantly greater amounts of time to cross the beam. Treatment of castrated males with EB significantly improved their performance. Performance of the castrated females was not changed by EB treatment and was similar to that observed with the castrated + EB males. Significant gender differences in dopamine concentrations (female > male) were obtained in the corpus striatum, as well as the olfactory tubercle and hypothalamus. Dopamine levels were unaltered by EB treatment. In Experiment II, behavioral and neurochemical determinations were directly compared between castrated and intact male mice. Behavioral performance of castrates was significantly reduced compared to intact males. No differences in dopamine concentrations were obtained between these two groups; however, the hypothalamic dopamine/DOPAC ratio of castrates was significantly greater than that of intact male mice. These results demonstrate significant modulatory effects of EB in castrated male, but not female, mice upon sensorimotor performance and indicate that this task may provide an effective means to partial out modulatory effects of gonadal steroid hormones upon skilled sensorimotor performance. When the data of Experiments I and II are combined, it appears that the basis of this sensorimotor deficit in the males is the absence of gonadal steroid hormones. In addition, these results reveal significant gender differences in various dopaminergic systems in these mice.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1994

12-lipoxygenases and 12(S)-HETE: role in cancer metastasis.

Arachidonic acid metabolites have been implicated in multiple steps of carcinogenesis. Their role in tumor cell metastasis, the ultimate challenge for the treatment of cancer patients, are however not well-documented. Arachidonic acid is primarily metabolized through three pathways, i.e., cyclooxygenase, lipoxygenase, and P450-dependent monooxygenase. In this review we focus our attention on one specific lipoxygenase, i.e., 12-lipoxygenase, and its potential role in modulating the metastatic process. In mammalian cells there exist three types of 12-lipoxygenases which differ in tissue distribution, preferential substrates, and profile of their metabolites. Most of these 12-lipoxygenases have been cloned and sequenced, and the molecular and biochemical determinants responsible for catalysis of specific substrates characterized. Solid tumor cells express 12-lipoxygenase mRNA, possess 12-lipoxygenase protein, and biosynthesize 12(S)-HETE [12(S)-hydroxyeicosatetraenoic acid], as revealed by numerous experimental approaches. The ability of tumor cells to generate 12(S)-HETE is positively correlated to their metastatic potential. A large collection of experimental data suggest that 12(S)-HETE is a crucial intracellular signaling molecule that activates protein kinase C and mediates the biological functions of many growth factors and cytokines such as bFGF, PDGF, EGF, and AMF. 12(S)-HETE plays a pivotal role in multiple steps of the metastatic 'cascade' encompassing tumor cell-vasculature interactions, tumor cell motility, proteolysis, invasion, and angiogenesis. The fact that 12-lipoxygenase is expressed in a wide diversity of tumor cell lines and 12(S)-HETE is a key modulatory molecule in metastasis provides the rationale for targeting these molecules in anti-cancer and anti-metastasis therapeutic protocols.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗

PubMed1994

The effect of reserpine treatment in vivo upon L-dopa and amphetamine evoked dopamine and DOPAC efflux in vitro from the corpus striatum of male rats.

In the present experiment we tested the effects of L-DOPA and amphetamine upon dopamine and DOPAC efflux in vitro from superfused corpus striatal tissue fragments of male rats who had been pretreated with reserpine. Male rats were treated with reserpine (5 mg/kg) or its vehicle at 24 hours prior to sacrifice and superfusion of the corpus striatum. Two different modes of L-DOPA (5 microM) and amphetamine (10 microM) stimulation, a brief 10-minute and a continuous 60-minute infusion, were tested for their ability to evoke striatal dopamine and DOPAC efflux. Depletion of monoamine storage capacity as achieved with reserpine significantly reduced the amount of basal dopamine and DOPAC released from superfused striatal tissue fragments of male rats. Although basal release rates were significantly reduced, the amount of dopamine and DOPAC released in response to in vitro L-DOPA infusions (10 or 60 minute infusions) was equivalent between reserpine and vehicle treated animals. In contrast, amphetamine stimulated DA release was significantly reduced in male rats treated with reserpine. For both L-DOPA and amphetamine, significantly greater amounts of dopamine were obtained with the 60- versus 10-minute infusion modes. These results demonstrate that the capacity for L-DOPA, but not amphetamine, to evoke dopamine efflux is unaltered under conditions when monoamine storage ability is diminished.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1994

Isolation and characterization of sets of anti-L-fucose and anti-bovine serum albumin antibodies directed against a glycoconjugate of L-fucose and bovine serum albumin.

A set of anti-carbohydrate antibodies and a set of anti-protein antibodies were isolated from the serum of rabbits immunized with a glycoconjugate of L-fucose and bovine serum albumin. The sets were separated by affinity chromatography by a two-column method on adsorbents with L-fucose or bovine serum albumin ligands. Isoelectrofocusing results showed that the anti-carbohydrate antibodies consisted of 11 molecular species and the anti-bovine serum albumin antibodies consisted of seven molecular species. The anti-carbohydrate antibodies are all of the IgG type while the anti-protein antibodies contain three types of globulin molecules, IgA, IgG, and IgM. The former antibodies should be useful as markers for unique glycoproteins of diseased cells and the latter antibodies may be useful for investigating the mechanism of simultaneous synthesis of three types of immunoglobulins.

Animals↗

PubMed1994

The effect of estrogen administration in vivo upon catecholamine release in vitro from superfused hypothalamic tissue of ovariectomized pre-pubertal and adult mice.

In the present report we examined the effects of estrogen upon catecholamine release from superfused medial basal hypothalamic tissue fragments of pre-pubertal ovariectomized CD-1 mice. Prepubertal mice treated with estradiol benzoate (EB--5 micrograms x 2 days, sc), showed significantly reduced amounts of dopamine but no changes in norepinephrine release in response to a depolarizing concentration of potassium (30 mmol/L) compared with their respective groups receiving the oil vehicle. Since EB treatment reduced potassium stimulated dopamine release in these pre-pubertal mice, in a second experiment we compared the effects of EB versus oil vehicle treatment upon potassium stimulated dopamine release from the hypothalamus of the ovariectomized adult female mouse. Similar to that observed in the pre-pubertal mouse, EB treatment significantly reduced the amount of potassium stimulated dopamine release. Interestingly, the absolute amounts of potassium stimulated dopamine release was substantially greater in adult compared with pre-pubertal mice. These results demonstrate that the hypothalamic dopaminergic system of both pre-pubertal and adult mice show relatively similar responses to estrogen treatment but differ in absolute amounts of dopamine released.

Aging↗

PubMed1994

Serotonergic involvement in the regulation of prolactin and vasoactive intestinal peptide mRNA expression in the rat anterior pituitary.

These studies examined the contribution of serotonin (5-HT) to the control of prolactin (PRL) and vasoactive intestinal peptide (VIP) messenger RNA expression in rat anterior pituitary. Daily injection of rats with the biosynthetic precursor to serotonin, 5-hydroxytryptophan (5-HTP; 25 mg/kg, q.i.d.), resulted on day 5 in a 50% increase in the expression of PRL mRNA in the pituitary while at the same time reducing the levels of both the 1.0 and 1.7 kb VIP mRNA transcripts. Co-treatment of rats with 5-HTP plus the catecholamine biosynthesis inhibitor, alpha-methyl-tyrosine (alpha-MT; 150 mg/kg, q.d. x 2 days), or the dopamine receptor antagonist haloperidol (1.25 mg/kg, b.i.d. x 5 days), resulted in increases in pituitary PRL message levels that were greater than those observed with either anti-dopaminergic agent alone. In contrast, 5-HTP was unable to reverse the inhibition of PRL mRNA expression caused by treatment with the dopamine receptor agonist bromocriptine (2.5 mg/kg, b.i.d. x 5 days). Neither alpha-MT, haloperidol nor bromocriptine had a significant effect on pituitary VIP mRNA expression. Administration of the direct-acting 5-HT receptor agonist quipazine (5 mg/kg, b.i.d.) for 14 consecutive days caused a significant increase in pituitary PRL mRNA levels on day 1 and reached a plateau of 90% above control levels on days 7 and 14. VIP mRNA levels rose significantly on day 1 of quipazine treatment but thereafter fell to a minimum of 22% (1.0 kb) and 52% (1.7 kb) of control by day 14.(ABSTRACT TRUNCATED AT 250 WORDS)

5-Hydroxytryptophan↗

PubMed1994

Neuroblastoma of parotid gland: report of a case and immunohistochemical characteristics.

A case of a parotid mass in a 2-year-old boy, postoperatively diagnosed as neuroblastoma, a rare tumour not previously reported in the parotid gland is presented. The neoplasm developed within the parotid gland as a painless mass without regional lymphadenopathy. Histopathologically, the tumour showed primitive nerve cells-neuroblasts-with round or oval dark basophilic nuclei and scanty cytoplasm. The cells were arranged in circular rosettes around an eosinophilic mass consisting of very fine filaments originating in the tumour cells or papillary configuration and sometimes scattered in the poorly developed stroma. Immunohistochemical evaluation of the tumour showed a positive immunoreactivity for vimentin, alpha and beta subunits of S-100 protein, neurone-specific enolase (NSE), substance P, met-enkephalin and chromogranin but cytokeratins, desmin, actin, myosin, glial fibrillary acidic protein (GFAP) and calcitonin gene related peptide (CGRP) were negative. The histopathological and immunohistochemical findings conclude a diagnosis of neuroblastoma of the parotid gland.

Child, Preschool↗

PubMed1994

Comparison of proliferating cell nuclear antigen index in benign and malignant salivary pleomorphic adenoma.

The expression of proliferating cell nuclear antigen (PCNA) was studied in benign and malignant pleomorphic adenomas by using monoclonal antibody to PCNA. Carcinoma in pleomorphic adenoma (n = 8), cell-rich variant (n = 6) and typical pleomorphic adenoma (n = 6) were selected in this study. The PCNA index in carcinoma in pleomorphic adenoma showed a higher index of nuclear staining (mean 22.9%, S.D. 6.2) than in typical pleomorphic adenoma (mean 6.9%, S.D. 3.4) or a cell-rich variant of pleomorphic adenoma (mean 8.8%, S.D. 3.3). A significant difference in PCNA index was found between benign and malignant pleomorphic adenoma (P < 0.05). The present study suggests that PCNA index significantly differs between pleomorphic adenoma and carcinoma in pleomorphic adenoma, but in the prediction of malignant transformation potential it should be combined with routine histopathological examination.

Adenoma, Pleomorphic↗

PubMed1994

Tissue reactions to titanium implants containing bovine bone morphogenetic protein: a scanning electron microscopic investigation.

Cylindric titanium implants treated with bovine bone morphogenetic protein (bBMP) (Group I) and with bovine serum albumin (BSA) (Group II) were implanted in the edentulous mandibles of 15 adult dogs. They were examined 1, 2, 4, 8, and 12 weeks after insertion of the implants by scanning electron microscopy (SEM). In all specimens from Group I, active bone formation had occurred between the implant and host bone, with a well-adapted fit between the implant and the newly formed bone 4 weeks after implantation. The surrounding bone was of normal structure by week 8. However, 3 months after implantation, fibrous tissue was consistently seen surrounding parts of the implants in Group II. The results of this study suggest that the implants treated with bBMP are capable of forming new bone of normal structure at a faster rate than BSA controls and that this new bone is closely adapted to the implant surface.

Animals↗

PubMed1994

Crystal structure of a tethered dimer of HIV-1 proteinase complexed with an inhibitor.

HIV-1 proteinase (HIV PR) is a dimeric enzyme composed of two identical polypeptide chains that associate with twofold symmetry. We have determined to 1.8 A the crystal structure of a covalently tethered dimer of HIV PR. The tethered dimer:inhibitor complex is identical in nearly every respect to the complex of the same inhibitor with the wild type dimeric molecule, except for the linker region. Our results suggest that the tethered dimer may be a useful surrogate enzyme for studying the effects of single site mutations on substrate and inhibitor binding as well as on enzyme asymmetry, and for simulating independent mutational drift of the two domains which has been proposed to have led to the evolution of modern day, single-chain aspartic proteinases.

Alcohols↗