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Biomedical subjects

Zhe Sun

Publications and source records attributed to Zhe Sun.

At least 19 recordsLinked to original sources

A truncated COL10A1 protein causes Schmid metaphyseal chondrodysplasia via protein downregulation and impairing α1 trimer formation and secretion.

Schmid-type metaphyseal chondrodysplasia (SMCD) is primarily caused by mutations in the COL10A1 gene. This study reports a novel frameshift mutation, c.1940dup (p.Asn647Lysfs*2), identified in a Chinese SMCD pedigree. The mutation did not alter messenger RNA levels but significantly reduced COL10A1 protein expression. The mutant protein lacks the C-terminal 33 amino acids, resulting in a truncated polypeptide of 648 residues with a lower molecular weight than the wild-type protein. Degradation kinetics analysis showed no evidence of accelerated turnover. Notably, even under complete inhibition of degradation pathways, mutant protein expression remained substantially lower than that of wild-type, suggesting a potential defect in translational efficiency. Furthermore, the mutation severely disrupted the assembly of the characteristic collagen X trimer and led to markedly reduced extracellular secretion, as measured by accumulated protein levels in conditioned medium. These findings demonstrate that the c.1940dup mutation contributes to SMCD pathogenesis through coordinated mechanisms involving protein truncation, reduced expression, probable translational deficiency, and defective trimer formation and secretion, thereby revealing new potential therapeutic targets.

Osteochondrodysplasias↗

Purification, properties and extended solution structure of the complex formed between human immunoglobulin A1 and human serum albumin by scattering and ultracentrifugation.

Immunoglobulin A (IgA) is unique amongst antibodies in being able to form polymeric structures that may possess important functions in the pathology of specific diseases. IgA also forms complexes with other plasma proteins, the IgA1-human serum albumin (HSA) complex (IgA1-HSA) being typical. We have purified this complex using a novel two-step purification based on thiophilic chromatography and gel filtration, and characterised this. HSA is linked covalently to the tailpiece of IgA1 by a disulphide bond between Cys471 in IgA1 and Cys34 in HSA. IgA1-HSA binds to IgA receptors on neutrophils and monocytes, and elicits a respiratory burst that is comparable in magnitude to that of monomeric IgA1. The solution arrangement of IgA1-HSA was identified by X-ray scattering and ultracentrifugation. The radius of gyration R(G) of 7.5(+/-0.3) nm showed that IgA1-HSA is more extended in solution than IgA1 (R(G) of 6.1-6.2 nm). Its distance distribution function P(r) showed two peaks that indicated a well-separated solution structure similar to that for IgA1, and a maximum dimension of 25 nm, which is greater than that of 21 nm for IgA1. Sedimentation equilibrium showed that the IgA1:HSA stoichiometry is 1:1. Sedimentation velocity resulted in a sedimentation coefficient of 6.4S and a frictional ratio of 1.87, which is greater than that of 1.56 for IgA1. The constrained modelling of the IgA1-HSA structure using known structures for IgA1 and HSA generated 2432 conformationally randomised models of which 52 gave good scattering fits. The HSA structure was located at the base of the Fc fragment in IgA1 in an extended arrangement. Such a structure accounts for the functional activity of IgA1-HSA, and supports our previous modelling analysis of the IgA1 solution structure. The IgA1-HSA complex may suggest the potential for creating a new class of targeted therapeutic reagents based on the coupling of IgA1 to carrier proteins.

Humans↗

Semi-extended solution structure of human myeloma immunoglobulin D determined by constrained X-ray scattering.

Human immunoglobulin D (IgD) occurs most abundantly as a membrane-bound antibody on the surface of mature B cells (mIgD). IgD possesses the longest hinge sequence of all the human antibody isotypes, with 64 residues connecting the Fab and Fc fragments. A novel rapid purification scheme of secreted IgD from the serum of an IgD myeloma patient using thiophilic (T-gel) and lectin affinity chromatography gave a stable, homogeneous IgD preparation. Synchrotron X-ray scattering and analytical ultracentrifugation of IgD identified the solution arrangement of its Fab and Fc fragments, and thereby its hinge structure. The Guinier X-ray radius of gyration R(G) of 6.9(+/-0.1)nm showed that IgD is more extended in solution than the immunoglobulin subclass IgA1 (R(G) of 6.1-6.2nm). Its distance distribution function P(r) showed a single peak at 4.7nm and a maximum dimension of 23nm. Velocity experiments gave a sedimentation coefficient of 6.3S, which is similar to that for IgA1 at 6.2S. The complete IgD structure was modelled using molecular dynamics to generate IgD hinge structures, to which homology models for the Fab and Fc fragments were connected. Good scattering curve fits were obtained with 18 semi-extended best fit IgD models that were filtered from 8500 trial models. These best-fit models showed that the IgD hinge does not correspond to an extended polypeptide structure. The averaged solution structure arrangement of the Fab and Fc fragments in IgD is principally T-shaped and flexible, with contribution from Y-shaped and inverted Y-shaped structures. Although the linear sequence of the IgD hinge is much longer, comparison with previous scattering modelling of IgA1 and IgA2(m)1 suggests that the hinge of IgA1 and IgD are more similar than might have been expected, Both possess flexible T-shaped solution structures, probably reflecting the presence of restraining O-linked sugars.

Amino Acid Sequence↗

Classification of breast microcalcifications: radiological-pathological correlation.

BACKGROUND: Microcalcifications play a very important role in detection of breast cancer, especially early stage breast cancer. However, ambiguity still exists in understanding the relationship between radiological and pathological characteristics of microcalcifications. The definitive indication of a biopsy has not been established. The purpose of this study is to evaluate the relationship of classification of breast microcalcifications using full-field digital mammography to the pathological characteristics. METHODS: For all the women an open biopsy had been conducted. One hundred and three mammographs showing clustered microcalcifications from 98 consecutive patients were reviewed along with their pathological records. To investigate the value of each criterion for the detection of cancer, univariate and multivariate analyses were performed on the entire sample and then on morphological subgroups. RESULTS: Pathological examination showed 67 malignant lesions (65.05%) and 36 benign lesions (34.95%). In the univariate analysis, four radiological variables were significant: morphological type (P = 0.001), complicated by a mass (P = 0.002), number of microcalcifications per cluster (P = 0.02) and linear or triangular distribution of clusters (P = 0.009). In the multivariate analysis, two criteria remained significant: morphological type (P < 0.001) and complicated by a mass (P = 0.001). The percentage of malignancy was 37.0%, 60.0%, 78.8%, and 88.9%, respectively, for type 2 (regularly punctiform), type 3 (dusty), type 4 (irregularly punctiform) and type 5 (vermicular) microcalcifications (Le Gal's classification). The malignancy was 78.6% for microcalcifications complicated by a mass and 48.9% without a mass. The difference was significant (P < 0.05). The relationship between morphological types of microcalcifications and the pathological characteristics was also studied. In subgroups, type 3 (dusty) microcalcifications complicated by a mass (P = 0.001) or with the number of microcalcifications more than 10 (P = 0.024); and type 2 (regularly punctiform) with a diameter of the area over 20 mm (P = 0.024) or complicated by a mass (P = 0.025) were statistically significant as criteria for malignant tumour. CONCLUSIONS: Most cases of microcalcifications of type 4 or 5; type 3 complicated by a mass or with the number of microcalcifications more than 10; type 2 complicated by a mass or with a diameter of the area over 20 mm; are indicative of cancer. Open biopsy is recommended to acquire definitive pathological diagnosis for these cases. For the remainder of the morphological types, stereotaxic biopsy or followup should be considered.

Adult↗

Mechanical properties of the interaction between fibronectin and alpha5beta1-integrin on vascular smooth muscle cells studied using atomic force microscopy.

The mechanical properties of integrin-extracellular matrix (ECM) interactions are important for the mechanotransduction of vascular smooth muscle cells (VSMC), a process that is associated with focal adhesions, and can be of particular significance in cardiovascular disease. In this study, we characterized the unbinding force and binding activity of the initial fibronectin (FN)-alpha5beta1 interaction on the surface of VSMC using atomic force microscopy (AFM). It is postulated that these initial binding events are important to the subsequent focal adhesion assembly. FN-VSMC adhesions were selectively blocked by antibodies against alpha5- and beta1-integrins as well as RGD-containing peptides but not by antibodies against alpha4- and beta3-integrins, indicating that FN primarily bound to alpha5beta1. A characteristic unbinding force of 39 +/- 8 pN was observed and interpreted to represent the FN-alpha5beta1 single-bond strength. The ability of FN to adhere to VSMC (binding probability) was significantly reduced by integrin antagonists, serum starvation, and platelet-derived growth factor (PDGF)-BB, whereas lysophosphatidic acid (LPA) increased FN binding. However, no significant change in the resolved unbinding force was observed. After engagement, the force required to dislodge the FN-coated bead from VSMC increased with increasing of contact time, suggesting a time-dependent increase in number of adhesions and/or altered binding affinity. LPA enhanced this process, whereas PDGF reduced it, suggesting that these factors also affect the multimolecular process of focal contact assembly. Thus AFM is a powerful tool for the characterization of the mechanical properties of integrin-ECM interactions and their regulation. Our results indicate that the functional activity of alpha5beta1 and focal contact assembly can be rapidly regulated.

Animals↗

Histamine effects on endothelial cell fibronectin interaction studied by atomic force microscopy.

Atomic force microscopy was used to investigate the cellular response to histamine, one of the major inflammatory mediators that cause endothelial hyperpermeability and vascular leakage. AFM probes were labeled with fibronectin and used to measure binding strength between alpha5beta1 integrin and fibronectin by quantifying the force required to break single fibronectin-integrin bonds. The cytoskeletal changes, binding probability, and adhesion force before and after histamine treatment on endothelial cells were monitored. Cell topography measurements indicated that histamine induces cell shrinkage. Local cell stiffness and binding probability increased twofold after histamine treatment. The force necessary to rupture single alpha5beta1-fibronectin bond increased from 34.0 +/- 0.5 pN in control cells to 39 +/- 1 pN after histamine treatment. Experiments were also conducted to confirm the specificity of the alpha5beta1-fibronectin interaction. In the presence of soluble GRGDdSP the probability of adhesion events decreased >50% whereas the adhesion force between alpha5beta1 and fibronectin remained unchanged. These data indicate that extracellular matrix-integrin interactions play an important role in the endothelial cell response to changes of external chemical mediators. These changes can be recorded as direct measurements on live endothelial cells by using atomic force microscopy.

Animals↗

Extended and flexible domain solution structure of the extracellular matrix protein anosmin-1 by X-ray scattering, analytical ultracentrifugation and constrained modelling.

Kallmann's syndrome corresponds to a loss of sense of smell and hypogonadotrophic hypogonadism. Defects in anosmin-1 result in the X-linked inherited form of Kallmann's syndrome. Anosmin-1 is an extracellular matrix protein comprised of an N-terminal, cysteine-rich (Cys-box) domain and a whey acidic protein-like (WAP) domain, followed by four fibronectin type III (FnIII) domains. The solution structures of recombinant proteins containing the first three domains (PIWF1) and all six domains (PIWF4) were determined by X-ray scattering and analytical ultracentrifugation. Guinier analyses showed that PIWF1 and PIWF4 have different radii of gyration (R(G)) values of 3.1 nm and 6.7 nm, respectively, but similar cross-sectional radii of gyration (R(XS)) values of 1.5 nm and 1.9 nm, respectively. Distance distribution functions showed that the maximum lengths of PIWF1 and PIWF4 were 11 nm and 23 nm, respectively. Analytical ultracentrifugation gave sedimentation coefficients of 2.52 S and 3.55 S for PIWF1 and PIWF4, respectively. The interpretation of the scattering data by constrained modelling requires homology models for all six domains in anosmin-1. While models were already available for the WAP and FnIII domains, searches suggested the Cys-box domain may resemble the cysteine-rich region of the insulin-like growth factor receptor. Automated constrained molecular modelling based on joining the anosmin-1 domains with structurally randomised linkers resulted in 10,000 models for anosmin-1. A trial-and-error search showed that about 0.1-1.4% of these models fitted the X-ray data. The best models showed that the three domains and six domains in PIWF1 and PIWF4, respectively, were extended. The inter-domain linkers in anosmin-1 could not all be extended at the same time, and there was evidence for inter-domain flexibility. Models with folded-back domain arrangements do not fit the data. These solution structures account for the known biological function of anosmin-1, in particular its ability to interact with its three macromolecular ligands.

Amino Acid Sequence↗

[Clinical value of computer aided detection system in direct full-field digital mammography: a preliminary evaluation].

OBJECTIVE: To study the clinical value of computer aided detection (CAD) in direct full-field digital mammography. METHODS: 480 digital mammograms of 120 patients with pathologically proved breast cancer were randomly selected and interpreted by an expert radiologist without knowing the pathologic diagnosis, then analyzed by CAD system, and finally read by the radiologist again to make the final diagnosis. The sensitivity for breast cancer detection of the radiologist in diagnosis of breast cancer was calculated before and after using CAD. Data of the breast density, diameters of mass and calcification, being spiculate or not, and pathology of each case were reviewed to determine how they impacted on the improvement of sensitivity with the assistance of CAD. RESULTS: The sensitivity for breast cancer detection by the radiologist before using CAD was 74.2%, significantly lower than that after using CAD (89.2%, chi(2) = 37.628, P = 0.000). Breast density (chi(2) = 7.352, P = 0.007), diameter of calcification (chi(2) = 10.549, P = 0.014) and pathologic pattern (chi(2) = 13.751, P = 0.003) were highly correlated with the improvement of sensitivity. Use of CAD might overcome type 1 missed diagnosis, and not type 2 missed diagnosis. CONCLUSION: Computer-aided detection helps radiologist improve their diagnostic accuracy of breast cancer, however, is not capable of differentiating benign from malignant lesions.

Adult↗

Cytological screening and 15 years' follow-up (1986-2001) for early esophageal squamous cell carcinoma and precancerous lesions in a high-risk population in Anyang County, Henan Province, Northern China.

BACKGROUND: Anyang County and nearby Linzhou (formerly Linxian) in Henan, Northern China have been well recognized as the highest incidence area for esophageal squamous cell carcinoma (SCC) in the world. SCC remains the leading cause of cancer-related death in these areas and the natural history for esophageal precancerous lesions is not clear. METHODS: In the present study, to determine the significance of cytological examination and natural history for SCC, esophageal balloon cytological screening and follow-up studies have been performed 2273 symptom-free subjects in Anyang County since 1986. Based on cellular morphological changes, esophageal epithelial cells were classified as normal (NOR), inflammation (IN), hyperplasia (HYP), dysplasia (grades I and II) (DYS I and DYS II), near squamous cell carcinoma (NSCC) and SCC. RESULTS: The prevalence of NOR, IN, HYP, DYS I, DYS II, and NSCC was 22.7, 3.6, 33.6, 21.5, 10.7, and 3.8%, respectively, in males and 25.8, 5.7, 33, 18.9, 10.4, and 3.8%, respectively in females. No difference was observed between the male and female subjects (p > 0.05). Of the 2273 subjects examined, 2199 subjects entered the final follow-up analysis (97%). During the 15 years' follow-up on these subjects, 94 new cases (4.2%, 94/2199), including 91 with SCC (97%, 91/94, 58 males and 33 females) and 3 with GCA (3%, 3/94, one male and two females) were identified with a mean time of 8 +/- 4.6 years after entry of the follow-up. The incidence for SCC in males was higher than that in females (p < 0.05). The rate for SCC development increased apparently from the groups of NOR (2.4%) to DYS I (5%), DYS II (8.3%), and NSCC (10.3%) (p < 0.001). The prevalence of DYS and NSCC increased significantly with age. CONCLUSIONS: The present results indicate that esophageal balloon cytology is a reliable approach for early SCC and precancerous lesions screening and that DYS and NSCC are important markers for high-risk subjects with predisposition to SCC. Close follow-up to the subjects with HYP, DYS and NSCC may shed light on the natural history for human esophageal carcinogenesis.

Adult↗

The dimeric and trimeric solution structures of the multidomain complement protein properdin by X-ray scattering, analytical ultracentrifugation and constrained modelling.

Properdin regulates the alternative pathway of the complement system of immune defence by stabilising the C3 convertase complex. It contains six thrombospondin repeat type I (TSR-1 to TSR-6) domains and an N-terminal domain. Properdin exists as either a dimer, trimer or tetramer. In order to determine the solution structure of multiple TSR domains, the molecular structures of dimeric and trimeric properdin were studied by X-ray scattering and analytical ultracentrifugation. Guinier analyses showed that the dimer and trimer have radii of gyration R(G) values of 7.5 nm and 10.3 nm, respectively, and cross-sectional radii of gyration R(XS) values of 1.3 nm and 1.5 nm, respectively. Distance distribution functions showed that the maximum lengths of the dimer and trimer were 25 nm and 30 nm, respectively. Analytical ultracentrifugation gave sedimentation coefficients of 5.1S and 5.2S for the dimer and trimer forms, respectively. Homology models for the TSR domains were constructed using the crystal structure of the TSP-2 and TSP-3 domains in human thrombospondin as templates. Properdin could be represented by seven TSR domains, not six as believed, since the crystal structure determined for TSP-2 and TSP-3 showed that the N-terminal domain (TSR-0) could be represented by a truncated TSR domain with the same six conserved Cys residues found in TSR-1 to TSR-6. Automated constrained molecular modelling revealed the solution conformations of multiple TSR domains in properdin at medium resolution. The comparison of 3125 systematically generated conformational models for the trimer with the X-ray data showed that good curve fits could be obtained by assuming that the linker between adjacent TSR domains possessed limited flexibility. Good trimer models correspond to partially collapsed triangular structures, and extended triangular shapes do not fit the data. The corresponding 3125 models for the dimer revealed a similar outcome in which a partially collapsed TSR structure gave good fits. The models account for the effect of mutations that cause properdin deficiencies, and suggest that the biologically active TSR-4, TSR-5 and TSR-6 domains are exposed for protein-protein interactions. The role of the other TSR domains in properdin may be to act as spacers to make TSR-4, TSR-5 and TSR-6 accessible for function.

Amino Acid Sequence↗

[Observations of the non-surgical treatment response on diabetic patients with chronic periodontitis].

PURPOSE: To observe the non-surgical treatment response on diabetic patients with chronic periodontitis. METHODS: Moderate to advanced chronic periodontitis was studied in 36 Diabetes Mellitus (DM) patients classified as 20 cases with high and fluctuating blood glucose level (DM-H) and 16 cases with relatively low and stable blood glucose level (DM-L). 28 non-DM patients with chronic periodontitis served as control (Non-DM). Plaque Index (PlI), Gingival Index (GI), Bleeding on Probing (BOP), Probing Depth (PD) and Clinical Attachment Loss (AL) of all patients were recorded at 6 sites on each tooth at the baseline and in the first, the third, the sixth month after oral hygiene instrument (OHI), scaling and root planing. RESULTS: It was found that the short-term effect of non-surgical periodontal procedure had resulted in significant resolution of gingival inflammation and pronounced reduction in pocket depth and gain of attachment loss in all patients. The treatment response was similar in both DM and Non-DM patients with chronic periodontitis. CONCLUSIONS: Non-surgical periodontal treatment allowed for favorable treatment responses in a group of diabetic patients with chronic periodontitis and that their various profiles of blood glucose did not influence the short-term healing response to the treatment.

Adult↗

[MVR-PCR and forensic medicine].

MVR mapping is one kind of DNA analytic techniques, which has developed greatly within 13 years. The molecular basis, history and applications in forensic medicine were systemically reviewed in this paper. Because of its umsubstitutive mole in forensic paternity test and individual identification, the technique is becoming one of the important methods of bioassay.

Base Sequence↗

Isolation and characterisation of mutants of GroEL that are fully functional as single rings.

A key aspect of the reaction mechanism for the molecular chaperone GroEL is the transmission of an allosteric signal between the two rings of the GroEL complex. Thus, the single-ring mutant SR1 is unable to act as a chaperone as it cannot release bound substrate or GroES. We used a simple selection procedure to identify mutants of SR1 that restored chaperone activity in vivo. A large number of single amino acid changes, mapping at diverse positions throughout the protein, enabled SR1 to regain its ability to act as a chaperone while remaining as a single ring. In vivo assays were used to identify the proteins that had regained maximal activity. In some cases, no difference could be detected between strains expressing wild-type GroEL and those expressing the mutated proteins. Three of the most active proteins where the mutations were in distinct parts of the protein were purified to homogeneity and characterised in vitro. All were capable of acting efficiently as chaperones for two different GroES-dependent substrates. All three proteins bound nucleotide as effectively as did GroEL, but the binding of GroES in the presence of ATP or ADP was reduced significantly relative to the wild-type. These active single rings should provide a useful tool for studying the nature of the allosteric changes that occur in the GroEL reaction cycle.

Adenosine Diphosphate↗

Cell-based and direct gene transfer-induced angiogenesis via a secreted chimeric fibroblast growth factor-1 (sp-FGF-1) in the chick chorioallantoic membrane (CAM).

Fibroblast growth factor-1 (FGF-1) is a potent angiogenic factor; its structure lacks a signal peptide for secretion. We previously reported that the overexpression of a secreted version of FGF-1 (sp-FGF-1) in microvascular endothelial cells (ECs) enhances cell migration [Partridge et al. J Cell Biochem 2000; 78(3): 487]. In the current study, we have examined the angiogenic effects of sp-FGF-1 in chicken chorioallantoic membranes (CAMs). Two methods of examining the effects of sp-FGF-1 in CAMs were used: cell-mediated transfection via bovine ECs and direct gene transfection. In the cell-mediated gene transfection, those eggs that were implanted with a gelatin sponge seeded with ECs stably transfected to over-express sp-FGF-1 protein showed a significant increase in angiogenesis inside the sponge when compared to eggs treated with vector control-transfected ECs. In the direct gene transfer, eggs received sp-FGF-1 showed a significant increase in vascularization when compared to eggs received vector alone plasmids. These CAM models are useful both for studying molecular mechanisms of angiogenesis and for developing better gene therapy strategies.

Allantois↗

[Effects of different class II cavity designs on stresses in restoration].

OBJECTIVE: To assess effects of different class II cavities on stresses of amalgam and composite resin restoration. METHODS: Five kinds of class II MO cavities were prepared on the first molars of mandibles. The cavities were filled by amalgam and composite resin, respectively. The stresses were analyzed by using three-dimensional finite element method, after vertical and lateral pressures were loaded on the teeth. RESULTS: The stresses were greater in traditional cavities than those in reformed cavities when 100 N vertical and lateral pressures were loaded on the teeth. But the pressing stress and shearing stress in cavities filled by amalgam were smaller in traditional cavities than those in reformed cavities. The stresses were smaller in cavities filled by resin than those in cavities filled by amalgam significantly, when the cavities were designed with slot retention groove, without slot retention groove and as plate form. The stresses were smaller in two kinds of slot and plate cavities than those in traditional and reformed cavities. CONCLUSION: It is better to design cavity with slot retention groove for amalgam restoration and to design cavity without slot retention groove for composite resin restoration.

Composite Resins↗

[Three dimensional finite element stress analysis in various class II cavity design on first maxillary premolar].

OBJECTIVE: To evaluate the stress influence in different contour of class II cavity with different filling materials on first maxillary premolar. METHODS: The stress distribution in two types and all three contours of class II cavity in maxillary premolar were analyzed by three dimensional finite element. The cavity was respectively filled with amalgam and composite resin; and the vertical and lateral stress force was calculated. RESULTS: In the different contours, the stress of composite restoration by slot design was close to that by dovetail design, and the stress of amalgam restoration by slot design, especially in vertical force, descended respectively at 41%,29% and 27%. CONCLUSION: When class II lesions is only limited in proximal region of the premolar, the filling of composite resin with slot design is recommended.

English Abstract↗

[Therapeutic Effect of rhIL-11 on Thrombocytopenia Induced by Carboplatin in Rhesus Monkeys]

The effectiveness of rhIL-11 on thrombocytopenia induced by carboplatin in rhesus monkeys was investigated. Thrombocytopenia was induced in monkeys by i.v. administration of carboplatin at a dose of 15 mg/kg(-1)/d(-1) for three consecutive days. rhIL-11 (50 or 100 micro g/kg(-1)/d(-1)) or Neumega (100 micro g/kg(-1)/d(-1)) were administered s.c. for 14 days beginning one day following the final dose of carboplatin. The results showed that rhIL-11 significantly improved mean platelet nadirs and shortened the mean duration of platelet counts less than 50% of pre-treatment values. Administration of rhIL-11 also resulted in moderate increase of the reticulated platelet, leukocyte and reticulocyte counts in peripheral blood and megakaryocytic and erythroid progenitors in bone marrow. rhIL-11 did not enhance ADP-induced platelet aggregation. These results indicate that rhIL-11 has a potent thrombopoietic effect in vivo and could be an important agent to reduce the severity and duration of thrombocytopenia following chemotherapy.

Journal Article↗

Hyperosmotic stress in murine hybridoma cells: effects on antibody transcription, translation, posttranslational processing, and the cell cycle.

Mechanisms for increased antibody production in batch cultures of murine hybridoma cells in response to hyperosmotic stress were investigated. The rates of immunoglobulin transcription and protein translation and posttranslational processing were determined in control and hyperosmotic cultures. Changes in immunoglobulin transcription played a minor role in the increase in antibody production in response to hyperosmotic stress. In contrast, protein translation increased substantially in response to osmotic stress. However, the antibody translation rate remained relatively constant after correcting for the overall increase in protein translation. Cell size and intracellular antibody pool also increased in response to hyperosmolarity. The intracellular antibody pool increased proportionately with the increase in cell size, indicating that hyperosmotic cultures do not selectively increase their intracellular antibody population. Changes in cell cycle distribution in response to osmotic stress and the relationship between the cell cycle and antibody production were also evaluated. Hyperosmotic stress altered the cell cycle distribution, increasing the fraction of the cells in S-phase. However, this change was uncorrelated with the increase in antibody production rate. Immunoglobulin degradation was relatively low ( approximately 15%) and remained largely unchanged in response to hyperosmotic stress. There was no apparent increase in immunoglobulin stability as a result of osmotic stress. Antibody secretion rates increased approximately 50% in response to osmotic stress, with a commensurate increase in the antibody assembly rate. The rate of transit through the entire posttranslational processing apparatus increased, particularly for immunoglobulin light chains. The levels of endoplasmic reticulum chaperones did not increase as a fraction of the total cellular protein but were increased on a per cell basis as the result of an increase in total cellular protein. A difference in the interactions between the immunoglobulin heavy chains and BiP/GRP78 was observed in response to hyperosmotic conditions. This change in interaction may be correlated with the decrease in transit time through the posttranslational pathways. The increase in the posttranslational processing rate appears to be commensurate with the increase in antibody production in response to hyperosmotic stress.

Animals↗