OBJECTIVETo compare the concentrations of IgA, lactate dehydrogenase, lysozyme and alkaline phosphatase (ALP) in unstimulated (UWS) and stimulated (SWS) whole saliva between children with severe early childhood caries (S-ECC) and children without caries.

METHODSOne hundred and ninety-two children aged from 42 to 54 months were recruited from 11 urban kindergartens in Beijing. The S-ECC group contained 98 children with more than 5 decayed teeth, and the control group contained 94 caries-free children. The age and sex were matched in the two groups. Two milliliter UWS and 2 ml SWS was collected between 9 and 11 a.m. The salivary IgA was measured by immunoturbidimetric technique. The concentrations of lactate dehydrogenase and ALP were measured by continuous monitoring method, while lysozyme was detected by turbidimetric technique. All results for paired observations between unstimulated and stimulated whole saliva were analysed by paired-samples t test.

RESULTSIn both UWS and SWS, the concentrations of IgA, lactate dehydrogenase and lysozyme in S-ECC children were higher than those in caries-free children (P < 0.01), but the concentration of ALP showed no significant difference in SWS between S-ECC children and caries-free children (P > 0.05).

CONCLUSIONSThe presence of early childhood caries may be associated with an increase of IgA, lactate dehydrogenase and lysozyme in unstimulated and stimulated whole saliva.

Alkaline Phosphatase ; analysis ; Child, Preschool ; Dental Caries ; metabolism ; Female ; Humans ; Immunoglobulin A ; analysis ; L-Lactate Dehydrogenase ; analysis ; Male ; Muramidase ; analysis ; Saliva ; enzymology ; immunology

AIMTo propose a new simple and sensitive voltammetric method for determination of proteins.

METHODSProtein with sulfhydryl or disulfide bond in 0.5 mol x L(-1) NaOH, 1.5 x 10(-4) mol x L(-1) Pb2+ and 0.02% tetrabutylammonium iodide was heated in boiling water for 5 minutes. The reactive product gave a well defined reductive adsorption wave at -0.66 V (vs SCE) by means of single sweep polarography, and the height of derivative wave was proportional to the concentration of proteins.

RESULTSThe peak height was linearly proportional to bovine serum albumin (BSA) or human serum albumin (HSA) concentration in range of 7.5 x 10(-10) -3.0 x 10(-7) mol x L(-1) (r(BSA) = 0.9995, and r(HSA) = 0.9990). The detection limit of BSA or HSA was 3.0 x 10(-10) mol x L(-1). For lysozyme (Lyso), the concentration range was from 1.4 x 10(-8) to 1.3 x 10(-6) mol x L(-10 (r(Lyso) = 0.9997) and the detection limit was 7.0 x 10(-9) mol x L(-1).

CONCLUSIONThe method is simple, rapid, sensitive and applicable to the assay of diluted human serum albumin samples.

Adsorption ; Animals ; Humans ; Lead ; Muramidase ; analysis ; Polarography ; methods ; Quaternary Ammonium Compounds ; Serum Albumin ; analysis ; chemistry ; Serum Albumin, Bovine ; analysis ; chemistry ; Sodium Hydroxide

OBJECTIVETo investigate the correlation between nurse job burnout and salivary lysozyme activity.

METHODSThe saliva samples of 131 subjects were collected at four time points for two consecutive days with saliva collection tubes. The acquisition time points were 8:00 (baseline concentration), 10:00 (morning), 15:30 (afternoon), and 17:30 (recovery period). At the same time every subjects completed the job burnout questionnaire to investigate their general demographic characteristics and job burnout level. The salivary lysozyme concentration was measured with ELISA. The data were analyzed by partial correlation analysis and multiple stepwise regression analysis.

RESULTSThere were significant differences in the salivary lysozyme activity between subjects with different ages, working years, and education levels. The work period vitality and the average energy of ≤ 30 age group were higher than other two groups and the recovery energy was higher than >35 age group. Working period vitality, the average energy of group >15 years were less than ≤ 10 years group. The work period energy and the average energy of university (college) and above group were lower than high school (secondary) and the following group. Job burnout and its three dimensions had a significant negative correlation with salivary lysozyme concentration (P < 0.01). Depersonalization and emotional exhaustion were the negative impact factors for salivary lysozyme activity at baseline. Emotional exhaustion and personal fulfillment were the negative impact factors for salivary lysozyme activity during the working period. Personal fulfillment was the negative factor for salivary lysozyme activity during the recovery period and the average salivary lysozyme activity.

CONCLUSIONSalivary lysozyme activity is sensitive for nurse job burnout, so it can be used as an objective evaluation index of job burnout.

Burnout, Professional ; epidemiology ; psychology ; Emotions ; Fatigue ; Humans ; Muramidase ; analysis ; Nurses ; psychology ; Occupational Diseases ; epidemiology ; psychology ; Regression Analysis ; Salivary Proteins and Peptides ; analysis ; Surveys and Questionnaires

No abstract available.
Actins/analysis ; Adult ; Female ; Human ; Immunohistochemistry ; Lactoferrin/analysis ; Muramidase/analysis ; Phosphopyruvate Hydratase/metabolism ; Pregnancy ; S100 Proteins/analysis ; Salivary Gland Neoplasms/chemistry/*pathology ; Salivary Glands/chemistry/*embryology ; Submandibular Gland/embryology ; alpha 1-Antitrypsin/analysis

Objective: To elucidate the possible role of human lysozyme-like protein 4 (LYZL4) in fertilization and characterize its enzymatic properties. METHODS: The localization of LYZL4 in human spermatozoa was investigated by immunofluorescence staining, the sources of LYZL4 on the sperm surface examined by RT-PCR, and the role of LYZL4 in fertilization assessed by the zona-free hamster egg penetration test. The recombinant plasmid pPIC9K-LYZL4 was constructed and its expression induced with methanol after transformed into competent Pichia pastoris GS115. The recombinant LYZL4 protein (rLYZL4) was purified from the fermentation supernatant and subsequently identified by Western blot. The hyaluronan binding ability of rLYZL4 was determined by ELISA and the muramidase activity, hyaluronidase activity, and free radical scavenging ability examined by spectrophotometric methods. RESULTS: Immunodetection with a specific antiserum localized LYZL4 on the acrosomal membrane of mature spermatozoa, which was exclusively secreted from the testis and epididymis as shown by RT-PCR. Immunoneutralization of LYZL4 significantly decreased the number of human spermatozoa bound to zona-free hamster eggs in a dose-dependent manner in vitro. The recombinant protein was expressed successfully by the P. pastoris strain GS115. Purified rLYZL4 exhibited a potent hyaluronan binding ability and a strong free radical scavenging ability but no muramidase or hyaluronidase activity. CONCLUSIONS LYZL4 secreted from the testis and epididymis is localized on the acrosomal membrane of mature spermatozoa and plays a role in sperm-egg binding as well as in binding hyaluronan and scavenging free radicals, which suggests that it might be a multi-functional molecule contributive to sperm protection and sperm-egg binding.
Acrosome ; enzymology ; Animals ; Blotting, Western ; Cricetinae ; Enzyme-Linked Immunosorbent Assay ; Epididymis ; Female ; Fertilization ; physiology ; Free Radical Scavengers ; metabolism ; Humans ; Hyaluronic Acid ; metabolism ; Male ; Muramidase ; analysis ; physiology ; Pichia ; Plasmids ; metabolism ; Recombinant Proteins ; analysis ; metabolism ; Sperm-Ovum Interactions ; physiology ; Spermatozoa ; enzymology ; Testis

PURPOSE: Pleural effusion, an accumulation of fluid in the pleural space, usually occurs in patients when the rate of fluid formation exceeds the rate of fluid removal. The differential diagnosis of tuberculous pleurisy and malignant pleural effusion is a difficult task in high tuberculous prevalence areas. The aim of the present study was to identify novel biomarkers for the diagnosis of pleural fluid using proteomics technology. MATERIALS AND METHODS: We used samples from five patients with transudative pleural effusions for internal standard, five patients with tuberculous pleurisy, and the same numbers of patients having malignant effusions were enrolled in the study. We analyzed the proteins in pleural fluid from patients using a technique that combined two-dimensional liquid-phase electrophoresis and matrix assisted laser desorption/ionization-time of flight-mass spectrometry. RESULTS: We identified a total of 10 proteins with statistical significance. Among 10 proteins, trasthyretin, haptoglobin, metastasis-associated protein 1, t-complex protein 1, and fibroblast growth factor-binding protein 1 were related with malignant pleural effusions and human ceruloplasmin, lysozyme precursor, gelsolin, clusterin C complement lysis inhibitor, and peroxirexdoxin 3 were expressed several times or more in tuberculous pleural effusions. CONCLUSION: Highly expressed proteins in malignant pleural effusion were associated with carcinogenesis and cell growth, and proteins associated with tuberculous pleural effusion played a role in the response to inflammation and fibrosis. These findings will aid in the development of novel diagnostic tools for tuberculous pleurisy and malignant pleural effusion of lung cancer.
Biomarkers* ; Carcinogenesis ; Ceruloplasmin ; Chaperonin Containing TCP-1 ; Clusterin ; Diagnosis ; Diagnosis, Differential ; Electrophoresis ; Fibroblasts ; Fibrosis ; Gelsolin ; Haptoglobins ; Humans ; Inflammation ; Lung Neoplasms ; Methods* ; Muramidase ; Pleural Effusion ; Pleural Effusion, Malignant ; Prevalence ; Proteomics ; Spectrum Analysis ; Tuberculosis ; Tuberculosis, Pleural

Nocardia cyriacigeorgica is an aerobic gram-positive rod that has mostly been reported as an opportunistic pathogen. Since molecular methodologies were introduced to identify species, infections caused by N. cyriacigeorgica have been reported. The patient was a 51-year-old woman with aplastic anemia, systemic lupus erythematosus, and disseminated tuberculosis, who was admitted to Chosun University Hospital with a history of fever and productive cough. During her hospitalization, sputum cultures were taken and a bacterium suspicious of acitinomycetes grew five times. It was a gram-positive rod that was also partially acid-fast on modified Kinyoun stain and resistant to lysozyme. After 24 h of incubation, cultures of the sputum onto sheep's blood agar plates (BAP) demonstrated rough, chalky, and white colonies with a characteristic earthy odor. Based on the above results, the presumptive identification of Nocardia species was made. To identify species of this isolate, 16S rRNA gene sequence analysis was taken and showed 99.9% homology to N. cyriacigeorgica DSM44484(T). The results of biochemical tests were compatible with other reports of N. cyriacigeorgica. As a result, this isolate was identified as N. cyriacigeorgica. Herein, we present a first report of N. cyriacigeorgica isolated from a patient with pulmonary infection in Korea.
Agar ; Anemia, Aplastic ; Cough ; European Continental Ancestry Group ; Female ; Fever ; Genes, rRNA ; Hospitalization ; Humans ; Korea ; Lupus Erythematosus, Systemic ; Middle Aged ; Muramidase ; Nocardia ; Odors ; Respiratory Tract Infections ; Sequence Analysis ; Sprains and Strains ; Sputum ; Tuberculosis

AIMTo investigate the inhibitory effect of egg white lysozyme (LZM) on ceftazidime (CFT)-induced release of endotoxin from Pseudomonas aeruginosa.

METHODSP. aeruginosa PAO1 was inoculated in nutrition broth or diluted rabbit blood free of antibiotics in the presence or absence of LZM and incubated at 37 degrees C on a water bath shaker. beta-Lactam antibiotic, CFT, was added to cultures at 3.5 h (nutrition broth culture) or 5 h (diluted rabbit blood culture) after inoculation. After 3 h of CFT treatment, the supernatants from different bacterial cultures were prepared by centrifuge and the concentrations of endotoxin in the supernatants were measured. The bacterial supernatants were also added to a murine macrophage cell line RAW 264.7 or intravenously injected into carrageenin-sensitized mice. Tumor necrosis factor-alpha (TNF alpha) and nitric oxide (NO) concentrations in RAW 264.7 supernatants or in mouse sera were tested.

RESULTSCFT treatment alone obviously inhibited the growth of P. aeruginosa PAO1 accompanied by strong and rapid bacteriolysis and released relatively high concentration of endotoxin from bacteria both in nutrition broth and in diluted rabbit blood cultures. The bacterial supernatant from CFT treatment alone yielded high concentrations of TNF alpha both in RAW 264.7 cells and in mice and high level of NO in RAW 264.7 cells. Treatment with the combination of LZM and CFT evidently blocked the lysis of bacteria and reduced the release of endotoxin without decreasing bactericidal activity of CFT. TNF alpha and NO productivity of the supernatants prepared from the LZM/CFT combinative treated bacterial cultures were significantly decreased both in RAW 264.7 cells and in mice indicating that the inflammatory activity was reduced.

CONCLUSIONLZM can effectively prevent CFT-induced bacteriolysis, endotoxin release and subsequent proinflammatory factor production but without decreasing bactericidal activity of CFT, resulting in the disassociation of bactericidal activity and bacteriolysis. Thus, LZM might be important for preventing endotoxemia in Gram-negative sepsis with the treatment of antibiotics.

Animals ; Bacteriolysis ; drug effects ; Ceftazidime ; pharmacology ; Egg White ; analysis ; Endotoxins ; metabolism ; Mice ; Muramidase ; isolation & purification ; pharmacology ; Nitric Oxide ; metabolism ; Pseudomonas aeruginosa ; metabolism ; physiology ; Rabbits ; Tumor Necrosis Factor-alpha ; metabolism

Objective: To prepare a polyclonal antibody against human lysozyme-like protein 4 (LYZL4) expressed in the prokaryotic system and identify the distribution of LYZL4 in the testis. METHODS: The full-length cDNA of LYZL4 was cloned into the pET32a plasmid and the expression of the recombinant LYZL4 (rLYZL4) was induced by IPTG. The rLYZL4 was purified by Ni-NTA and chitin affinity chromatography respectively and its bactericidal activity was observed by bilayer agar plate diffusion assay. The purified rLYZL4 was used as an immunogen to generate the polyclonal antibody, followed by examination of the antibody titer by ELISA and its specificity by Western blot. The distribution of LYZL4 in human tissue, sperm and seminal plasma was identified and its subcellular localization in the testis was determined by immunohistochemistry. RESULTS: rLYZL4 was expressed efficiently in the prokaryotic system and exhibited no bacteriolytic activity against M. lysodeikticus and E. coli. The anti-rLYZL4 polyclonal antibody could bind the recombinant protein with a high sensitivity and specificity. LYZL4 was identified in the testis, epididymis and sperm protein extracts and localized in the acrosomal region of round and elongating spermatids. CONCLUSIONS An anti-rLYZL4 polyclonal antibody was successfully prepared using the prokaryotic expression system. LYZL4 was detected in the acrosomal region of round and elongating spermatids, suggesting an association with the structure and function of the acrosome.
Acrosome ; immunology ; Animals ; Antibodies ; analysis ; Blotting, Western ; DNA, Complementary ; Enzyme-Linked Immunosorbent Assay ; Epididymis ; immunology ; Escherichia coli ; Humans ; Immunohistochemistry ; Male ; Muramidase ; genetics ; immunology ; Plasmids ; Recombinant Proteins ; genetics ; Semen ; immunology ; Spermatozoa ; immunology ; Testis ; immunology

BACKGROUND AND OBJECTIVES: Mucin hypersecretion is one of the main symptoms of inflammatory diseases in the respiratory tract. The authors previously reported that pleiotypic pro-inflammatory cytokine, interleukin (IL)-1beta, plays significant roles in the respiratory tract inflammation by inducing mucins (MUC2, MUC5AC, MUC8). However, the molecular mechanism for mucin hypersecretion in the respiratory tract is still unclear. MATERIALS AND METHOD: In order to understand the mechanisms of mucin hypersecretion in the airway epithelium, the differentially expressed proteins and genes in the lung mucoepidermoid carcinoma cell line (NCI-H292 cells), which were treated for 6 and 24 hours with IL-1beta (10 ng/ml), were identified using 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE) proteomics and cDNA microarray analysis (8.6 K). RESULTS: In the 2-D PAGE, 8 differentially expressed proteins and 14 post-translational modification proteins were identified 6 and 24 hrs after the IL-1beta treatment. Microarray analysis identified a total of 413 genes (6.6%) in the 6-hour treatment group and 115 genes (2.0%) in the 24-hour treatment group that were regulated after the IL-1beta treatment. The differentially expressed genes that were regulated by the IL-1beta treatment were mostly found in the metabolic pathway rather than in the regulatory pathway. A comparison of the proteomic and microarray data showed that there was a large discrepancy between the protein expression and the gene expression levels. Among the genes encoding the proteins secreted in the airway, MUC5B was down-regulated but sialomucin CD 164, lysozyme, and the secretory leukocyte protease inhibitor (SLPI) were up-regulated. CONCLUSION: These results clearly show that the transcript levels have little value in predicting the extent of protein expression. Genomics and proteomics have different evaluation fields. Therefore, they may not provide all the information on the gene and protein profiles.
Carcinoma, Mucoepidermoid ; Cell Line ; Electrophoresis, Polyacrylamide Gel ; Epithelial Cells* ; Epithelium ; Gene Expression ; Genomics ; Inflammation ; Interleukin-1beta* ; Interleukins ; Lung ; Metabolic Networks and Pathways ; Microarray Analysis ; Mucins ; Mucus ; Muramidase ; Oligonucleotide Array Sequence Analysis ; Protein Processing, Post-Translational ; Proteomics ; Respiratory System ; Secretory Leukocyte Peptidase Inhibitor ; Sialomucins