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Biomedical subjects

H An

Publications and source records attributed to H An.

At least 19 recordsLinked to original sources

Compartmentalization of the matrix formed by nucleus pulposus and annulus fibrosus cells in alginate gel.

Intervertebral disc cells cultured in alginate gel are capable of reforming in alginate, a matrix that consists of two compartments: a rim of metabolically active cell-associated matrix and a more abundant, but metabolically less active, further removed matrix. At any one age and in most species, the cell-associated matrix formed by a nucleus pulposus or annulus fibrosus cell cultured in this way is less abundant than that formed by an articular chondrocyte. In both the cell-associated matrix and further removed matrix, the ratio of aggrecan to collagen is significantly higher in the case of nucleus pulposus than of annulus fibrosus, a feature that also distinguishes the matrices of the nucleus pulposus and annulus fibrosus in vivo. Nucleus pulposus and annulus fibrosus cells from older donors show a decreased ability to reform a cell-associated matrix rich in aggrecan. There is, however, some evidence that gene therapy and/or exposure of the cells to defined stimulatory factors can help overcome some of these age-related limitations. This contention is supported by recent evidence that nucleus pulposus and annulus fibrosus cells from adult donors can be manipulated to form, using the recently developed alginate-recovered chondrocyte system, a resilient tissue that bears many of the characteristics of the tissue in which these cells reside in vivo.

Alginates↗

Synthesis of novel 3'-C-methylene thymidine and 5-methyluridine/cytidine H-phosphonates and phosphonamidites for new backbone modification of oligonucleotides.

Novel 5'-O-DMT- and MMT-protected 3'-C-methylene-modified thymidine, 5-methyluridine, and 5-methylcytidine H-phosphonates 1-7 with O-methyl, fluoro, hydrogen, and O-(2-methoxyethyl) substituents at the 2'-position have been synthesized by a new effective strategy from the corresponding key intermediates 3'-C-iodomethyl nucleosides and intermediate BTSP, prepared in situ through the Arbuzov reaction. The modified reaction conditions for the Arbuzov reaction prevented the loss of DMT- and MMT-protecting groups, and directly provided the desired 5'-O-DMT- and/or MMT-protected 3'-C-methylene-modified H-phosphonates 1-6 although some of them were also prepared through the manipulation of protecting groups after the P-C bond formation. The modified Arbuzov reaction of 3'-C-iodomethyl-5-methylcytidine 53, prepared from its 5-methyluridine derivative 42, with BTSP provided the 5-methylcytidine H-phosphonate 54, which was further transferred to the corresponding 4-N-(N-methylpyrrolidin-2-ylidene)-protected H-phosphonate monomer 7. 5'-O-MMT-protected 3'-C-methylene-modified H-phosphonates 5, 3, and 7 were converted to the corresponding cyanoethyl H-phosphonates 50, 51, and 56 using DCC as a coupling reagent. One-pot three-step reactions of 50, 51, and 56 provided the desired 3'-C-methylene-modified phosphonamidite monomers 8-10. Some of these new 3'-methylene-modified monomers 1-10 have been successfully utilized for the synthesis of 3'-methylene-modified oligonucleotides, which have shown superior antisense properties including nuclease resistance and binding affinity to the target RNA.

Anti-Infective Agents↗

Quantitative measurements of cerebral blood flow in patients with unilateral carotid artery occlusion: a PET and MR study.

Although it has been demonstrated that quantitative measures of cerebral blood flow (CBF) can be obtained with the singular value decomposition (SVD) algorithm, the extent to which quantitative CBF measurements can be utilized under pathophysiological conditions has not been systematically studied. A total of five healthy volunteers and five patients with unilateral carotid artery occlusion were studied. Only magnetic resonance (MR) images were acquired for the volunteer group while both MR and positron emission tomography (PET) images were acquired for the patient group. Assessments of CBF from normal volunteers compared favorably with values reported in the literature. However, while a linear relationship was observed for each patient when MR measured CBF was compared to that obtained from PET, this linear relationship diminished when all patients were analyzed as a group (r = 0.41). A correction factor (CF) was proposed that was equal to the ratio of the area of the venous output function (VOF) in each patient to the mean VOF obtained from the volunteer group. After globally scaling the CBF of each patient based on the experimentally derived CF, a substantial improvement was observed (a slope of 1.02 and r = 0.8 for the linear regression line) in the relationship between MR estimated CBF and those obtained from PET.

Analysis of Variance↗

Cathepsin L: a predominant heat-activated proteinase in arrowtooth flounder muscle.

Characterization of the autolytic profile of arrowtooth flounder (ATF) muscle indicated the involvement of heat-activated proteinases active at both acidic and alkaline pH values. Further assay of fish extract exhibited the maximum activity at 60 degrees C against casein used as a substrate at both pH 5.5 and 8.0. The maximum activity shifted to lower temperatures by the addition of urea with two distinctive patterns: activity reduction at pH 5.5 and activity enhancement at pH 8.0. The highest inhibition by E-64 indicated the proteinase belongs to the cysteine proteinase class. At pH 5.5, the proteinase hydrolyzed Z-Phe-Arg-NMec and all types of protein substrates tested at higher rate than that at pH 8.0. Activity bands, observed on the activity-stained substrate gels, indicated similar proteinases are responsible for the proteolytic activity observed at both pH values. When proteins of fish extract were separated by HPLC-SEC, only one proteolytic peak was observed at the retention time of 26 min with an estimated molecular weight of 39800 Da. The results implied cathepsin L is a predominant proteinase responsible for autolysis of ATF muscle at elevated temperatures.

Animals↗

Phage display selection of hairpin loop soyacystatin variants that mediate high affinity inhibition of a cysteine proteinase.

Two hairpin-loop domains in cystatin family proteinase inhibitors form an interface surface region that slots into the active site cleft of papain-like cysteine proteinases, and determine binding affinity. The slot region surface architecture of the soybean cysteine proteinase inhibitor (soyacystatin N, scN) was engineered using techniques of in vitro molecular evolution to define residues that facilitate interaction with the proteinase cleft and modulate inhibitor affinity and function. Combinatorial phage display libraries of scN variants that contain mutations in the essential motifs of the first (QVVAG) and second (EW) hairpin-loop regions were constructed. Approximately 1010-1011 phages expressing recombinant scN proteins were subjected to biopanning selection based on binding affinity to immobilized papain. The QVVAG motif in the first hairpin loop was invariant in all functional scN proteins. All selected variants (30) had W79 in the second hairpin-loop motif, but there was diversity for hydrophobic and basic amino acids in residue 78. Kinetic analysis of isolated scN variants identified a novel scN isoform scN(LW) with higher papain affinity than the wild-type molecule. The variant contained an E78L substitution and had a twofold lower Ki (2.1 pM) than parental scN, due to its increased association rate constant (2.6 +/- 0.09 x 107 M-1sec-1). These results define residues in the first and second hairpin-loop regions which are essential for optimal interaction between phytocystatins and papain, a prototypical cysteine proteinase. Furthermore, the isolated variants are a biochemical platform for further integration of mutations to optimize cystatin affinity for specific biological targets.

Base Sequence↗

Identification of bacteria crucial to histamine accumulation in pacific mackerel during storage.

Bacterial growth and histamine formation in Pacific mackerel during storage at 0, 4, 15, and 25 degrees C were monitored. To identify bacterial species contributing to histamine formation, several groups of bacteria were isolated by using selective media under temperatures corresponding to the various storage conditions. Initially, low counts of bacteria were found in the gill, skin, and intestine of fresh fish, and only weak histamine formers were found in the gill. Histamine was found in the muscle when fish were stored above 4 degrees C, and aerobic plate counts reached 10(6) CFU/g. When fish became unsuitable for human consumption by abusive storage, toxicological levels of histamine were always found. The highest level of histamine formed was 283 mg/100 g in 2 days. The optimum temperature for supporting growth of prolific histamine formers was 25 degrees C. The most prolific and prevalent histamine former was Morganella morganii, followed by Proteus vulgaris, both of which were isolated on violet red bile glucose (VRBG) agar. At 15 degrees C, a significant level of histamine was still produced in fish muscle, although prolific histamine formers were less frequently detected than at 25 degrees C. The isolates on thiosulfate citrate bile salts sucrose (TCBS) agar were weak histamine formers and identified as Vibrio parahaemolyticus and Vibrio alginolyticus. At 4 degrees C, less than 57.4 mg/100 g of histamine was found in fish stored for 14 days. Most isolates were natural bacterial flora in the marine environment and identified as weak histamine formers. At 0 degrees C, neither histamine former nor histamine production was detected up to 14 days of storage.

Agar↗

Source and identification of histamine-producing bacteria from fresh and temperature-abused albacore.

Histamine-producing bacteria were isolated from fresh and temperature-abused albacore using two different isolation procedures. Typically, the bacterial isolates on Niven's or modified Niven's medium produced negligible or low levels of histamine (<300 ppm) in histamine enumeration broth. The most frequently found species using this approach was Hafnia alvei. By prescreening on selective media (eosin methylene blue [EMB] agar for enteric bacteria; deMan Rogosa Sharpe agar for lactic acid bacteria: KF streptococcus agar for streptococci; pseudomonas isolation [PI] agar for pseudomonads; and staphylococcus medium 110 agar for staphylococci) prior to plating on histidine decarboxylase differential media, detection rate of true histamine formers increased. Prolific histamine producers capable of forming >1,000 ppm histamine in culture broth were isolated when PI and EMB agars were used for prescreening. Among the selective media tested, EMB agar was most effective in selecting high histamine producers, as demonstrated by the highest rate of true positives based on histamine analysis. Histamine-producing isolates were mostly enteric bacteria, including Morganella morganii, H. alvei, Klebsiella spp., Citrobacter freundii, Enterobacter spp., and Serratia spp. M. morganii isolated on PI agar from temperature-abused albacore muscle was found to be the highest histamine former. This species was not isolated from fresh albacore. while other enteric bacteria were frequently detected on the gills. However, only a few species isolated from both fresh and temperature-abused muscles were identified as high histamine formers.

Agar↗

[Wet synthesis and characterization of fluoride-substituted hydroxyapatite].

Hydroxyapatite and fluoride-substituted hydroxyapatite powders are synthesized by wet method, compacted into pellets and subjected to heat treatment at 300, 600, 900 degrees C respectively. Analysis of the fluoride-substituted hydroxyapatites and HA by FTIR spectroscopy indicated that with the increasing of sintering temperatures, the phase-pure fluoride-substituted hydroxyapatite and stoichiometric hydroxyapatite could be got. According to the X-ray diffraction analysis, the fluoride-substituted hydroxyapatite is less stable for decomposing into beta-Ca3(PO4)2 at the temperature of 900 degrees C. The in vitro bioactivity of the fluoride-substituted hydroxyapatites and hydroxyapatite heat treated at 900 degrees C was assessed by immersion testing in simulated body fluid (SBF). Both scanning electron microscopy (SEM) analysis and ionic solution changes (Ca2+) demonstrated that the low fluoride content apatite could induce a surface bonelike apatite layer formation.

Fluorides↗

[Effects of supplementing of calcium, iron and zinc on women's health during pregnancy].

OBJECTIVES: To determine the effects of supplementing biscuits fortified with calcium (Ca) and vitamin D (VD), iron (Fe), vitamin C (VC) and zinc (Zn) to pregnant women from the 5th month of gestation until delivery on their health, and to explore a way to improve their Ca, Fe and Zn nutritional status during pregnancy. METHODS: A total of 313 healthy and primary pregnant women were enrolled and divided into five study groups based on their order visiting the hospitals for prenatal care. Each woman of the study groups was given three pieces of biscuit fortified with VD, Ca and VD, Ca, Zn and VD (Ca + Zn + VD), Ca, Fe, VC and VD (Ca + Fe + VD), Ca, Fe, VC, Zn and VD (Ca + Fe + Zn + VD), respectively, from the 15th month of gestation until delivery (24 weeks in total) daily. The fortified levels were 10 micrograms VD, 400 mg Ca from carbonate calcium, 10 mg Zn from lactate zinc, 10 mg Fe from ferrous lactate, and 50 mg VC, respectively. A parturient women was selected from the same hospital as control after one trial subject for each study groups selected. RESULTS: The daily dietary intakes of Ca, Zn and Fe in pregnant women were only 47.7%, 54.7% and 86.7% of the Recommended Dietary Allowances for Chinese. Incidence of anemia for mid-term pregnant women was 35.2%, and Fe-supplementation could significantly improved their hemoglobin level (P < 0.05). Prevalence of anemia in the groups of Ca + Fe + Zn + VD and Ca + Fe + VD was 35.3% and 40.7%, respectively, before Fe supplementation and reduced to zero and 4.0%, respectively, after Fe supplementation. Whereas, prevalence of anemia in the other groups without Fe supplementation still kept in a relatively high level. In the groups supplemented with Ca, their plasma Ca level increased, especially with the best results in Ca + VD group. Plasma level of Zn declined with length of gestation, which could be improved by Zn supplementation. Serum level of alkaline phosphatase activity increased a little bit with length of gestation. There was no significant difference in radial and ulnar bone mineral density (BMD) between trial groups and controls two months after delivery. Maternal radial and ulnar BMD correlated significantly with their dietary Ca intakes in Ca + VD group. CONCLUSION: The best way to improve maternal nutritional status is supplementation of Ca + Fe + Zn + VitD, based on the Recommended Dietary Allowances for Chinese.

Adult↗

[Effects of supplementing calcium, iron and zinc on the fetus development and growth during pregnancy].

OBJECTIVES: The purpose of the present study is to determine the effects of supplementing the biscuits fortified with calcium (Ca), iron (Fe) and zinc (Zn) for pregnant women from the 5th month of gestation until delivery on the growth and development of fetus and to explore the best way to improve infant growth. METHODS: A total of 313 healthy and primary pregnant women were enrolled and divided into five study groups based on their order visiting the hospitals for prenatal care. Each woman of the study groups was given three pieces of biscuit fortified with VD, Ca and VD, Ca, Zn and VD (Ca + Zn + VD), Ca, Fe, VC and VD (Ca + Fe + VD), Ca, Fe, VC, Zn and VD (Ca + Fe + Zn + VD), respectively, from the 5th month of gestation until delivery (24 weeks in total) daily. The fortified levels were 10 micrograms Ca from carbonate calcium, 10 mg Zn from lactate zinc, 10 mg Fe from ferrous lactate, and 50 mg VC, respectively. A parturient women was selected from the same hospital as control after one trial subject for each study groups selected. Placenta was weighed and umbilical cord venous blood and placenta tissue samples were collected after delivery for measuring Ca and other trace elements. Body weight and height of the new-born babies were measured. RESULTS: The weight of placenta in control group (551.1 +/- 64.2) g was significantly (P < 0.05) heavier than other trial groups with Ca + Fe + Zn + VD (467.1 +/- 36.6) g, but contents of Ca and other trace elements in placenta of the control group were lower, which probably because of the lower nutrients intake of mothers in control group that would result in a compensation enlargement of placenta to get enough nutrients for fetus from mother. The Hb level in cord venous blood was significantly higher than that of maternal blood. There was no significant difference in activity of alkaline phosphatase between cord venous blood and material blood. The contents of Ca, Fe and Zn in cord venous plasma were all significantly higher than those in maternal plasma. Birth weight and height in Ca + Fe + Zn + VD group were the highest in all the groups, with birth weight of (3.53 +/- 0.33) kg, as compared with (3.28 +/- 0.54) kg in the control group. There was significant correlation between birth weight, weight for age and intake of Ca, Fe and Zn during pregnancy. CONCLUSION: Supplementation of Ca + Fe + Zn + VitD during pregnancy is the best way to improve infant development and growth, based on the current Recommended Dietary Allowances (RDA) for pregnant women.

Adult↗

[The diagnosis and treatment of isolated sphenoid sinus disease].

OBJECTIVE: To detail the underlying symptoms and signs of patients with isolated sphenoid sinus inflammatory disease and enhance the initial diagnostic accuracy. METHOD: 49 patients with isolated sphenoid sinus inflammatory disease were reviewed, and the outcomes of the endoscopic surgery were compared with the non-endoscopic surgery. RESULT: In patients who underwent endoscopic sphenoidotomy, 31 were virtually free from symptoms and 5 still experienced mild headache and hyposmia but their radiographic finding had become normalized. None of them need re-operation within 6-48 months follow-up. However, in patients with non-endoscopic sinus irrigation, only one was free from the symptoms and 5 patients underwent re-operation because of recurrence. CONCLUSION: Nasal endoscopy and CT/MRI scan could provide a precise diagnosis for isolated sphenoid sinus disease and endoscopic surgery is valuable in its treatment.

Adolescent↗

[Observations of the curative effect with various liquid for post operative irrigation of ESS of treating chronic sinusitis and nasal polyps].

OBJECTIVE: To explore the influence of liquid for post-operative irrigation on the effect of ESS. METHOD: 192 cases with chronic sinusitis and/or nasal polyps of groupIIstageII who had undergone the ESS from September, 1998 to June, 1999 and completed a 3-month follow-up were divided randomly into 3 groups to accept 3 different kinds of irrigation liquid. The evaluation of curative effect was according to the index of HaiKou ESS-97. RESULT: The curative effect of Herba Houttuyniae group was better than the other two. CONCLUSION: The post-operative care is important and the irrigation liquid can affect the effect of ESS.

Adolescent↗

[Functional and morphological observation on cells in allergic rhinitis guinea-pigs with cold or heat syndrome].

OBJECTIVE: To investigate the functional and morphological characteristics of cells in allergic rhinitis (AR) guinea-pigs with Cold or Heat Syndrome. METHODS: The Cold Syndrome or Heat Syndrome guinea-pig model was made, and then reformed into AR model or rhinitis medicamentosa (RM) model by using nose drops of 10% toluene di-isocyanate (TDI) or 0.1% naphazoline hydrochloride. The cell function, count and ultramicroscopic morphology characteristics in the model animals were observed. RESULTS: The degranulation index (DI), of blood basophils and the count of mast cells (MC) in nasal mucosa were higher in AR guinea-pigs with Cold Syndrome than that in Heat Syndrome guinea-pigs, but the two parameters in AR with both Cold and Heat Syndrome were higher than those in the RM model. Mast cell and eosinophilic granulocyte in AR guinea-pigs with Cold Syndrome were high in count, widely infiltrated, indiscernible cellular capsule, heterogeneous in density, and vacuoles, fusions and tubules in granules while in those with Heat Syndrome, mast cells and eosinophils were scarcely found, without infiltration, granules with discernible membrane and homogeneous in density, no vacuoles, fusions and tubules in granules. CONCLUSION: The basophilic cells, mast cells and eosinophilic granulocytes in AR guinea-pigs with Cold Syndrome are in activated condition.

Animals↗

Evolution of sporadic olivopontocerebellar atrophy into multiple system atrophy.

OBJECTIVE: To determine the percentage of sporadic olivopontocerebellar atrophy (sOPCA) patients who later develop multiple system atrophy (MSA). METHODS: Observations of the course of 51 sOPCA patients 20 years of age or older initially evaluated in an ataxia clinic over 14 years and followed at 3- to 6-month intervals for 3 months to 10 years (median 2.5 years, interquartile range 5 months to 4 years). RESULTS: Seventeen patients evolved to develop MSA, whereas the remaining 34 manifested only progressively worsening cerebellar ataxia. The features of the MSA cases included autonomic failure and parkinsonism in 10 patients, autonomic failure without parkinsonism in six, and parkinsonism without autonomic failure in one. Using survival analysis methods, the authors estimated that 24% of subjects in this population will evolve to MSA within 5 years of the onset of sOPCA symptoms (95% CI 10% to 36%). An older age at onset of symptoms and a shorter time from onset of symptoms to first presentation in a neurology specialty clinic were both highly predictive of evolution to MSA. Six of the 17 patients who evolved to MSA died 4 months to 5 years after they had met diagnostic criteria for MSA. The estimated median survival time from time of transition was 3.5 years. In contrast, death occurred in only one of the 34 patients with sOPCA who did not evolve to MSA. Autopsy examination of all six patients with MSA who died confirmed the diagnosis. CONCLUSIONS: Approximately one-fourth of sporadic olivopontocerebellar atrophy patients will evolve to multiple system atrophy within 5 years, and this transition carries a poor prognosis for survival. Older age at onset of ataxia and earlier presentation in a neurologic specialty clinic predicted transition to MSA.

Adult↗

Presence of the LEE (locus of enterocyte effacement) in pig attaching and effacing Escherichia coli and characterization of eae, espA, espB and espD genes of PEPEC (pig EPEC) strain 1390.

In the present study, attaching and effacing Escherichia coli (AEEC) O45 isolates from post-weaning pigs with diarrhoea were examined for the presence of the LEE (locus of enterocyte effacement) using various DNA probes derived from the LEE of human enteropathogenic E. coli (EPEC) strain E2348/69. The LEE fragment was conserved among the eae -positive pig isolates. The attaching and effacing activity of PEPEC (pig EPEC) O45 isolates is highly correlated with the presence of the LEE. Nevertheless, for some PEPEC isolates, the insertion site of the LEE is different or has diverged during evolution. The presence of the LEE fragment in PEPEC isolates provides further evidence that the LEE region is conserved among AEEC of different animal origins. In addition, the nucleotide sequence of the region containing the eae gene and esp genes of a pig AEEC isolate, strain 1390, was determined. Among examined Eae proteins, Eae of strain 1390 showed the highest similarity with Eae belonging to the beta intimin group such as the Eae of rabbit AEEC. Moreover, all pig strains that produced attaching and effacing lesions in piglets and pig ileal explants belonged to the beta intimin group. The deduced amino acid sequences of the EspA, EspB and EspD proteins of strain 1390 showed particularly strong homology to those of AEEC strains presenting a beta intimin allele. Thus, pig AEEC possess the LEE sequences, and for the strain 1390, sequences of the eae and esp regions are related to those of other AEEC, in particular, strains presenting a beta intimin allele, such as the rabbit AEEC.

Adhesins, Bacterial↗

Rainbow trout (Oncorhynchus mykiss) cystatin C: expression in Escherichia coli and properties of the recombinant protease inhibitor.

Cystatins are a superfamily of low Ki cysteine proteinase inhibitors found in both plants and animals. Cystatin C, a secreted molecule of this family, is of interest from biochemical and evolutionary points of view, and also has biotechnological applications. Recently we cloned and sequenced the cDNA for rainbow trout (Oncorhynchus mykiss) cystatin C [Li et al., 1998. Molecular cloning, sequence analysis and expression distribution of rainbow trout (Oncorhynchus mykiss) cystatin C. Comp. Biochem. Phys. B 121, 135-143]. To explore the relationship between protein structure and function of trout cystatin C, we established a bacterial system for expression of the protein. Trout cystatin C expressed in the cytoplasm of bacterial cells did not have detectable protease inhibitory activity. Activity was regained by Ni-NTA chromatography under denaturing conditions followed by dialysis-based refolding. Titration of purified cystatin C preparations with papain indicated that approximately 20% of the total protein had been converted to the active form after one refolding cycle. Expression levels were 3-5 mg/l. The protease-inhibitory properties of recombinant trout cystatin C were similar to those of human and chicken cystatin C derived from biological sources and recombinant cystatin C derived from rat and mouse genes. The Ki for papain was 1.2 x 10(-15) M, exhibiting the high affinity binding unique to this family of protease inhibitors.

Animals↗

Physicochemical changes and mechanism of heat-induced gelation of arrowtooth flounder myosin.

Physicochemical changes of myosin during heating were investigated to elucidate the mechanism of heat-induced gelation of arrowtooth flounder (ATF) myosin at high ionic strength. Changes in dynamic properties indicated ATF myosin formed a gel in three different stages as shown by the first increase in G' (storage modulus) at 28 degrees C, followed by the decrease at 35 degrees C and the second increase at 42 degrees C. DSC thermogram showed the onset of myosin denaturation at 25 degrees C with two maximum transition temperatures at 30 and 36 degrees C. The decrease in alpha-helical content indicated ATF myosin began to unfold at 15 degrees C and the unfolding continued until it reached 65 degrees C. Turbidity measurement showed myosin began to aggregate at 23 degrees C and the aggregation was complete at 40 degrees C. Surface hydrophobicity increased consistently in the temperature range studied, 20-65 degrees C. Sulfhydryl contents decreased significantly at 20-30 degrees C due to the formation of disulfide linkages but remained constant at temperatures >30 degrees C. ATF myosin was shown to be extremely sensitive to heat, resulting in denaturation at lower temperature than other fish myosin. Denaturation was initiated by unfolding of the alpha-helical region in myosin followed by exposure of hydrophobic and sulfhydryl residues, which are subsequently involved in aggregation and gelation processes.

Animals↗

Effects of proteolysis and mechanism of gel weakening in heat-induced gelation of fish myosin.

Addition of papain decreased the onset temperature and the rate at which G' developed during heat-induced gelation of arrowtooth flounder myosin. Frequency sweep results revealed that G' markedly decreased in proportion to the amount of papain added. However, use of E-64, a cysteine proteinase inhibitor, reversed the effects of papain and protected myosin heavy chain from degradation. DSC thermograms indicated papain significantly decreased the enthalpy required to induce myosin denaturation without significant changes in onset and maximum transition temperatures. Thermal denaturation kinetics indicated decreases in both the activation energy and the rate of myosin denaturation. CD studies revealed a rapid decrease in alpha-helical content, indicating the initial degradation of myosin molecules mostly occurred in the tail region. These results suggested that proteolysis affected thermal properties and reactivity of myosin during heating. Although myosin gel could be formed, structural disruption resulted in lower gelling ability and rigidity of the formed gel.

Animals↗