Backbone resonance assignments for the Fv fragment of the catalytic antibody NPN43C9 with bound p-nitrophenol.
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Animals↗
Biomedical subjects
J Chung
Publications and source records attributed to J Chung.
Assignment of 1H, 13C, and 15N resonances of reduced Escherichia coli glutaredoxin 2.
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Bacterial Proteins↗
Expression and characterization of anti-NCA-95 scFv (CEA 79 scFv) in a prokaryotic expression vector modified to contain a Sfi I and Not I site.
The CEA 79 antibody has been used in bone marrow scintigraphy for the differential diagnosis of skeletal tumors and the evaluation of the bone marrow status of patients with various hematological disorders. The specific localization of radio-labeled CEA 79 antibody in bone marrow depends on its reactivity with NCA-95 (nonspecific cross-reacting antigen-95) present on the surface and in the cytosol of human granulocytes and myelopoietic cells. To make a CEA 79 scFv molecule that would be less immunogenic and more penetrating than the intact mouse immunoglobulin, we constructed a pRSET Sfi I/Not I expression vector. The scFv gene was then excised from a pCANTAB 5 E phage display vector by digestion with Sfi I and Not I and inserted into the pRSET Sfi I/Not I expression vector. Upon transformation of a BL21(DE3)pLysS strain of E. coli, CEA 79 scFv became expressed in inclusion bodies requiring a renaturation process for solubilization. The final yield of CEA 79 scFv was 5 mg per a liter of culture. The refolded CEA 79 scFv exhibited an affinity (Kd = 2.1 x 10(-9) M) equivalent to that of the original CEA 79 antibody (K(d) = 3.3 x 10(-9) M) and the same immunoreactivity to CEA and NCA-95 in Western blots and in immunohistochemical staining experiments.
Amino Acid Sequence↗
Effect of Na-hyaluronan on stromal and endothelial healing in experimental corneal alkali wounds.
The healing of stroma and endothelium after a standardized corneal alkali wound was evaluated in sodium-hyaluronan (Na-HA)-treated eyes. Alkali wounds were produced in one eye of each rabbit by applying a 5.5-mm round filter paper soaked in 1 N NaOH onto the central cornea for 60 s. Eyes were then treated with either 1% Na-HA (the treatment group) or phosphate-buffered saline (the control group) 4 times per day for 3 weeks. Endothelial wound morphometry was performed after alizarin red and trypan blue staining, while stromal healing was assessed by counting polymorphonuclear leukocytes (PMNs) and keratocytes in the central and marginal wound areas. During the early healing period the stroma treated with Na-HA had less PMNs than that of the control group. The size of the endothelial defect area measured 5 days after injury was significantly smaller in the Na-HA group than in the control corneas. The present findings indicate that topically applied 1% Na-HA affects stromal and endothelial healing during the early repair process after corneal alkali wounds.
Alkalies↗
Sclerosing Mucoepidermoid carcinoma with eosinophilia of the thyroid glands: a case report with clinical manifestation of recurrent neck mass.
Sclerosing mucoepidermoid carcinoma with eosinophilia (SMECE) is a recently recognized malignant neoplasm of the thyroid gland. About 14 cases of SMECE have been reported and this is the first reported case in Korea. A 57-year-old woman presented with right neck mass for 20 years. Total thyroidectomy was performed under the impression of thyroid carcinoma. The resected thyroid gland showed a poorly circumscribed hard mass. Histologically, the tumor consisted of solid nests of large atypical cells with dense fibrous stroma. The tumor cells showed squamoid appearance with abundant eosinophilic cytoplasm. There were also rare mucin-containing cells within the nests. Within the hyalinized stroma, numerous eosinophils were found. The surrounding thyroid parenchyma displayed Hashimoto's thyroiditis. There was metastasis in a regional lymph node. Two years after initial surgery, she underwent a modified radical neck dissection due to recurrent neck mass. After the radiation therapy for eight weeks, laryngectomy and esophagectomy were performed due to a recurrent carcinoma in the esophageal wall. We report an additional case of SMECE, with metastasis to regional lymph nodes and esophagus. The tumor appears to be more aggressive than previously reported and a correct diagnosis can be rendered by just examining the metastatic lesions.
Carcinoma, Mucoepidermoid↗
Flow cytometric immunophenotyping in fine-needle aspiration of lymph nodes.
Fine-needle aspiration (FNA) of lymph nodes has been regarded as a useful method in the diagnosis of lymphadenopathy. However, this procedure has been shown to be of limited value in the diagnosis of low or intermediate grade malignant lymphomas in some studies. Immunophenotyping is an essential adjunct to cytomorphology for the diagnosis of lymphoma by FNA. Immunophenotyping using flow cytometry (FCM) is rapid, objective and reliable. Using FCM, multiparametric analysis of 33 FNA materials from lymph nodes was performed and profiles of surface markers of lymphoid cells were assessed. In reactive hyperplasia, patterns of cell surface markers were quite variable, but disclosed polyclonality. Most of the B-cell lymphomas showed immunophenotypes for B-cell lineages with their kappa: lambda or lambda: kappa ratio being over 3:1. In T-cell lymphomas, T-cell surface markers were predominantly expressed as well. In conclusion, our results suggest that immunophenotyping of lymph node aspirates is a valuable diagnostic adjunct for lymphoproliferative disorders, particularly in B-cell lymphomas because immunophenotyping can be easily and adequately performed by FCM.
Antigens, CD19↗
Complete hydatidiform mole. A disease with a changing profile.
OBJECTIVE: To retrospectively evaluate the clinical presentation of complete molar pregnancies in an academic primary obstetrics and gynecology practice over the past decade. STUDY DESIGN: All cases of abnormal pregnancy presenting to our institution during the first half of gestation were identified through a computerized database. Clinical presentation and course of complete moles were analyzed. RESULTS: Twenty-four complete molar pregnancies were identified among 2,431 abnormal early gestations (1%). The patients' mean age was 24.5 years, and the mean gestational age was 9.5 weeks of amenorrhea (range, 8-25). Seventy-five percent of the patients presented with vaginal bleeding and 54% with excessive uterine size. None had hyperemesis gravidarum, preeclampsia, clinical hyperthyroidism or ovarian enlargement. All patients had abnormally elevated serum beta-hCG. Transvaginal ultrasound was diagnostic in more than half the patients, while it was suggestive of the diagnosis in the remainder. One patient experienced postevacuation trophoblastic embolization and developed persistent gestational trophoblastic disease. CONCLUSION: Due to the routine use of transvaginal ultrasound and serum beta-hCG in the workup of early gestational abnormalities, complete molar pregnancy rarely presents today with the traditional signs and symptoms. Despite their absence, the potential for persistent trophoblastic disease still exists, and careful follow-up is warranted.
Adult↗
Polyclonality and multispecificity of the CTL response to a single viral epitope.
The molecular anatomy of an immunodominant, Ld restricted CTL epitope located between residues 28-39 in hepatitis B surface Ag was defined to explore the immunologic constraints on mutational escape from the CTL response during a viral infection. Using a panel of hepatitis B surface Ag residue 28-39-specific CTL clones, the response to this epitope was found to be extremely diverse at the level of TCR fine specificity and beta-chain usage. Although each clone recognized shared as well as unique residues within the epitope as TCR contact sites, even the shared residues were recognized differently by different TCRs. Despite these differences, all clones were comparably cytolytic following Ag stimulation and produced similar amounts of antiviral cytokines previously shown to inhibit HBV replication. These results demonstrate that the CTL response to individual viral epitopes can be markedly polyclonal and multispecific, such that mutational inactivation of a single TCR contact site will not usually lead to viral escape from all CTL clones of the same epitope specificity. Given these constraints and the fact that the CTL response is usually directed against several different epitopes during most viral infections, mutational inactivation of a single epitope is not likely to be sufficient to cause viral persistence.
Amino Acid Substitution↗
Chronic alcohol intake reduces retinoic acid concentration and enhances AP-1 (c-Jun and c-Fos) expression in rat liver.
Chronic ethanol intake may interfere with retinoid signal transduction by inhibiting retinoic acid synthesis and by enhancing activator protein-1 (AP-1) (c-Jun and c-Fos) expression, thereby contributing to malignant transformation. To determine the effect of ethanol on hepatic retinoid levels, retinoic acid receptors (RARs) and AP-1 (c-Jun and c-Fos) gene expression, chronic ethanol (36% of total calorie intake) pair-feeding was conducted on rats for a 1-month period. Retinoic acid, retinol, and retinyl ester concentrations in both liver and plasma were examined by using high-performance liquid chromatography (HPLC). Both retinoic acid receptor (alpha, beta, gamma) and AP-1 (c-Jun and c-Fos) expression in the rat liver were examined by using Western blot analysis. Treatment with high-dose ethanol led to a significant reduction of retinoic acid concentration in both the liver and the plasma (11- and 8.5-fold reduction, respectively), as compared with animals pair-fed an isocaloric control diet containing the same amount of vitamin A. Similar to the retinoic acid reductions, both retinol and retinyl palmitate levels in the livers of the alcohol-fed group decreased significantly, but in smaller fold reduction (6.5- and 2.6-fold reduction, respectively). Ethanol did not modulate the expression of RARalpha, -beta, and -gamma genes in the liver. However, chronic alcohol feeding enhanced AP-1 (c-Jun and c-Fos) expression by 7- to 8-fold, as compared with the control group. These data suggest that functional downregulation of RARs by inhibiting biosynthesis of retinoic acid and up-regulation of AP-1 gene expression may be important mechanisms for causing malignant transformation by ethanol.
Animals↗
Function of the human interleukin 4 receptor (IL-4R)-derived acidic motif revealed by cytoplasmic domain chimeras of the IL-4R alpha chain and the IL-2R beta chain.
Interleukin 2 (IL-2)- and IL-4-mediated stimulation of survival and growth, reflected by the induction of bcl2 and c-myc, respectively, depends on the integrity of the membrane-proximal region (S-region) in the IL-2 receptor beta-chain (IL-2R beta) and the haematopoietin homology box1-containing region of the IL-4 receptor alpha-chain (IL-4R alpha). In contrast to IL-4, IL-2 induces the expression of c-fos and c-jun family genes, mediated by the acidic region (A-region) within the cytoplasmic domain of IL-2R beta. A highly acidic motif is also present in IL-4R alpha, and evidence in favour and against its importance has been published. The authors have constructed chimeric receptors between IL-2R beta and IL-4R alpha by substitution of either the S-region or the A-region of IL-2R beta with sequences derived from IL-4R alpha. These chimeras were stably transfected into BA/F3 cells and assayed for the capacity to restore functions of IL-2 beta, such as growth mediation by IL-2 and the induction of proto-oncogenes (c-myc, c-junB and c-fos). Replacement of both the S- and A-region of IL-2R beta with IL-4R alpha derived regions of similar size and cytoplasmic location supported growth-stimulation by IL-2 as well as proto-oncogene induction. In contrast, all IL-2R functions were lost by exchange of the S-region with the corresponding part of IL-4R alpha. Induction of c-junB and c-fos RNA as an indicator of A-region function, however, was maintained in an IL-2R beta chimera containing the acidic box-bearing region of IL-4R alpha. These data indicate a functional role of the acidic region in the IL-4R alpha-chain.
Animals↗
Chemical shift as a probe of molecular interfaces: NMR studies of DNA binding by the three amino-terminal zinc finger domains from transcription factor IIIA.
We report the NMR resonance assignments for a macromolecular protein/DNA complex containing the three amino-terminal zinc fingers (92 amino acid residues) of Xenopus laevis TFIIIA (termed zf1-3) bound to the physiological DNA target (15 base pairs), and for the free DNA. Comparisons are made of the chemical shifts of protein backbone 1HN, 15N, 13C alpha and 13C beta and DNA base and sugar protons of the free and bound species. Chemical shift changes are analyzed in the context of the structures of the zf1-3/DNA complex to assess the utility of chemical shift change as a probe of molecular interfaces. Chemical shift perturbations that occur upon binding in the zf1-3/DNA complex do not correspond directly to the structural interface, but rather arise from a number of direct and indirect structural and dynamic effects.
Amino Acid Sequence↗
The establishment of active chromatin domains.
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Animals↗
Oral fluids as an alternative to serum for measurement of markers of immune activation.
Oral fluids are convenient alternatives to blood sampling for evaluating significant metabolic components. Two forms of oral fluids, oral mucosal transudates (OMT) and saliva, were collected and compared for content of soluble products of immune activation. The data confirm that OMT and saliva represent distinct body fluids. The concentrations, outputs, and analyte/protein ratios of beta-2-microglobulin (beta2M), soluble tumor necrosis factor alpha receptor II (sTNFalphaRII), and neopterin were measured. Both the OMT and the saliva of most of the individuals in the control healthy populations had measurable levels of all three activation markers. When the immune system is activated, as in human immunodeficiency virus (HIV) infection, the levels of beta2M and sTNFalphaRII are increased in both OMT and saliva compared to those in a healthy control population. OMT levels correlated better with levels in serum than did saliva and appear to reflect systemic immune activation in HIV infection. Because acquisition of oral fluids is noninvasive and easily repeatable, measurement of beta2M and/or sTNFalphaRII content in OMT could be useful in the assessment of disease activity in patients with HIV infection or chronic inflammatory diseases.
AIDS-Related Opportunistic Infections↗
Role of orlistat in the treatment of obese patients with type 2 diabetes. A 1-year randomized double-blind study.
OBJECTIVE: Obesity is an important risk factor for type 2 diabetes. Weight loss in patients with type 2 diabetes is associated with improved glycemic control and reduced cardiovascular disease risk factors, but weight loss is notably difficult to achieve and sustain with caloric restriction and exercise. The purpose of this study was to assess the impact of treatment with orlistat, a pancreatic lipase inhibitor, on weight loss, glycemic control, and serum lipid levels in obese patients with type 2 diabetes on sulfonylurea medications. RESEARCH DESIGN AND METHODS: In a multicenter 57-week randomized double-blind placebo-controlled study, 120 mg orlistat or placebo was administered orally three times a day with a mildly hypocaloric diet to 391 obese men and women with type 2 diabetes who were aged > 18 years, had a BMI of 28-40 kg/m2, and were clinically stable on oral sulfonylureas. Changes in body weight, glycemic control, lipid levels, and drug tolerability were measured. RESULTS: After 1 year of treatment, the orlistat group lost 6.2 +/- 0.45% (mean +/- SEM) of initial body weight vs. 4.3 +/- 0.49% in the placebo group (P < 0.001). Twice as many patients receiving orlistat (49 vs. 23%) lost > or = 5% of initial body weight (P < 0.001). Orlistat treatment plus diet compared with placebo plus diet was associated with significant improvement in glycemic control, as reflected in decreases in HbA1c (P < 0.001) and fasting plasma glucose (P < 0.001) and in dosage reductions of oral sulfonylurea medication (P < 0.01). Orlistat therapy also resulted in significantly greater improvements than placebo in several lipid parameters, namely, greater reductions in total cholesterol, (P < 0.001), LDL cholesterol (P < 0.001), triglycerides (P < 0.05), apolipoprotein B (P < 0.001), and the LDL-to-HDL cholesterol ratio (P < 0.001). Mild to moderate and transient gastrointestinal events were reported with orlistat therapy, although their association with study withdrawal was low. Fat-soluble vitamin levels generally remained within the reference range, and vitamin supplementation was required in only a few patients. CONCLUSIONS: Orlistat is an effective treatment modality in obese patients with type 2 diabetes with respect to clinically meaningful weight loss and maintenance of weight loss, improved glycemic control, and improved lipid profile.
Adult↗
Thrombspondin acts via integrin-associated protein to activate the platelet integrin alphaIIbbeta3.
Integrin-associated protein (IAP or CD47) is a receptor for the cell/platelet-binding domain (CBD) of thrombospondin-1 (TS1), the most abundant protein of platelet alpha granules. Although it associates with alphaIIbbeta3, IAP has no known function in platelets. TS1, the CBD, and an IAP agonist peptide (4N1K) from the CBD of TS1 activate the platelet integrin alphaIIbbeta3, resulting in platelet spreading on immobilized fibrinogen, stimulation of platelet aggregation, and enhanced tyrosine phosphorylation of focal adhesion kinase. Furthermore, 4N1K peptide selectively stimulates the phosphorylation of LYN and SYK and their association with FAK. The phosphorylation of SYK is blocked by pertussis toxin, implicating a Gi-like heterotrimeric G protein. IAP solublized from membranes of unstimulated platelets binds specifically to an affinity column of 4N1K peptide. Both alphaIIb and beta3 integrin subunits and c-Src bind along with IAP. This complex of proteins is also detected with immunoprecipitation. Activation of platelets with the agonist peptide 4N1K results in the association of FAK with the IAP-alphaIIbbeta3 complex. Thus an important function of TS1 in platelets is that of a secreted costimulator of alphaIIbbeta3 whose unique properties result in its localization to the platelet surface and the fibrin clot.
Amino Acid Sequence↗
Upper limb work-related musculoskeletal disorders among newspaper employees: cross-sectional survey results.
At a metropolitan newspaper office in Canada with extensive video display terminal (VDT) use, researchers carried out a survey (n = 1,007, 84% response) to establish baseline prevalence of work-related musculoskeletal disorders (WMSDs) and to identify demographic, postural, task, and psychosocial factors associated with WMSD symptoms. One-fifth of the respondents reported moderate or worse upper limb pain recurring at least monthly or lasting more than a week over the previous year. Logistic regression showed that employees who faced frequent deadlines and high psychological demands (fast work pace and conflicting demands), had low skill discretion and social support, spent more time keyboarding, or who had their screen in a non-optimal position were more likely to report moderate to severe symptoms. Women reported significantly higher levels of symptoms than men.
Adult↗
Hepatitis C virus core and E2 protein expression in transgenic mice.
Transgenic mice have been produced that express the hepatitis C virus (HCV) core protein in the liver under the transcriptional control of the mouse major urinary protein promoter. These animals express the full length core protein in cytoplasm of their hepatocytes at levels comparable to those detected in naturally infected patients, without histological or biochemical evidence of liver disease or hepatocellular carcinoma. This contrasts with recent reports that HCV core protein can transform NIH 3T3 cells and cooperates with H-ras to transform primary rat fibroblasts in vitro. Coexpression of HCV core protein in double transgenic mice that replicate the hepatitis B virus (HBV) does not inhibit hepatocellular HBV gene expression or replication, contrary to reports that it inhibits HBV replication in HuH-7 cells after transient transfection in vitro. We have also produced transgenic mice in which a C-terminally truncated (aa384-715) glycosylated HCV E2 protein is expressed in the liver under the transcriptional control of the mouse albumin promoter. Despite the high level expression of HCV E2 protein, no evidence of liver disease was detected in these animals. These results suggest that the HCV core and E2 proteins are not cytopathic for the hepatocyte in vivo, and they represent an initial step in the development of a small animal model of HCV immunopathology.
Animals↗
A comparison of surgeons' posture during laparoscopic and open surgical procedures.
BACKGROUND: There is increasing recognition of surgeons' physical fatigue in the new ergonomic environment of laparoscopic surgery. The purpose of this study was to determine what the differences are in the movement of the surgeon's axial skeleton between laparoscopic and open operations. METHODS: Surgeons' body positions were recorded on videotape during four laparoscopic (LAP) and six open (OP) operations. The percent of time the head and back were in a normal, bent, or twisted position as well as the number of changes in head and back position were tabulated using a computer program. A separate laboratory study was performed on four surgeons "walking" a 0.5-inch polyethylene tubing forward and backward using laparoscopic and open techniques. The movements of the surgeons' head, trunk, and pelvis were measured using a three-camera kinematic system (Kin). The center of pressure was recorded using a floor-mounted forceplate (Fp). RESULTS: In the operating room surgeons' head and back positions were more often straight in laparoscopic procedures and more often bent in open operations. The number of changes in back position per minute were significantly decreased when the laparoscopic-only part of surgery was analyzed. In the laboratory the subjects' head position was significantly (p = 0.02) more upright and the anteroposterior (AP) and rotational range of motion of the head was significantly reduced during laparoscopy. Subjects' CP was more anterior and there was a significant reduction in the AP range of motion of the CP during laparoscopy. CONCLUSIONS: Our study suggests that surgeons exhibit decreased mobility of the head and back and less anteroposterior weight shifting during laparoscopic manipulations despite a more upright posture. This more restricted posture during laparoscopic surgery may induce fatigue by limiting the natural changes in body posture that occur during open surgery.
Ergonomics↗