Copper deficit as a potential pathogenic factor of reduced bone mineral density and severe tooth wear.

UNLABELLED: The study evaluated if men and women with severe tooth wear were at increased risk of general bone loss. Enamel biopsies obtained from 50 subjects aged 47.5 ± 5 years showed decreased copper content, which was associated with reduced spine bone mineral density, suggesting deficits of this trace element contributing to bone demineralization, enamel attrition, and deteriorated quality of mineralized tissues. INTRODUCTION: The objective of this cross-sectional study was to assess associations between enamel trace minerals and bone mineral density (BMD) in severe tooth wear. We hypothesized that similar factors contributed to both the excessive abrasion of dental enamel and reduced BMD in subjects with tooth wear. METHODS: Fifty patients aged 47.5 ± 5 years with severe tooth wear and 20 age-, sex-, and body mass index (BMI)-matched healthy volunteers with normal dental status were studied regarding dietary intakes of trace elements, serum and salivary copper (Cu), zinc (Zn), and calcium (Ca) concentrations, and serum PTH, osteocalcin, and hydroxyvitamin D levels. Tooth wear was determined using clinical examination based on standard protocol according to Smith and Knight. In all subjects, acid biopsies of the maxillary central incisors were carried out to assess mineral composition of the enamel. Atomic absorption spectroscopy with an air/acetylene flame was used to measure Ca and Zn, and graphite furnace atomic absorption spectroscopy was used to analyze Cu content. BMD was examined using dual energy X-ray absorptiometry. RESULTS: Tooth wear patients had reduced lumbar spine, but not femoral, BMD relative to controls (p < 0.001). No differences were found in enamel Ca concentration and Zn content was slightly higher in tooth wear patients than in controls whereas Cu content was significantly decreased in the patients: 19.59 ± 16.4 vs 36.86 ± 26.1 µg/l (p = 0.01) despite similar levels of Cu in serum and saliva. The differences were independent of serum 25-OH-D, osteocalcin concentrations or PTH either. CONCLUSION: Severe tooth wear is associated with reduced spinal BMD. Enamel in adult individuals with severe tooth wear is low in copper content. Therefore, further work is needed to determine whether copper plays a role in bone pathophysiology in these patients.

Carbohydrate-deficient transferrin depends on disease activity in rheumatoid arthritis and systemic sclerosis.

OBJECTIVES: Changes in the glycosylation of plasma proteins have been linked to the aetiology of the rheumatic diseases. The aim of this study was to determine and compare the levels of carbohydrate-deficient transferrin (CDT) in patients with rheumatoid arthritis (RA), systemic sclerosis (SSc), and systemic lupus erythematosus (SLE). METHOD: Studies were carried out in 29 female patients with RA, 27 with SSc, and 17 with SLE. CDT was assayed by the N Latex CDT immunonephelometric assay. RESULTS: The levels of %CDT in the sera of RA, SLE, and SSc patients were significantly higher than in controls while the absolute concentrations of CDT were unchanged. %CDT, CDT, and transferrin do not differ significantly between patients with rheumatic diseases. In RA and SSc patients, a positive correlation was observed between %CDT and C-reactive protein (CRP), as well as a positive correlation in RA patients between %CDT and 28-joint Disease Activity Score (DAS28). CONCLUSIONS: The changes in the serum %CDT concentration in patients with RA and SSc correlated with disease activity markers.

Higher importance of interleukin 6 than classic tumor markers (carcinoembryonic antigen and squamous cell cancer antigen) in the diagnosis of esophageal cancer patients.

It has been suggested that interleukin 6 (IL-6) plays a potential role in the growth and progression of tumors, including esophageal cancer (EC). The aim of the study was to compare clinical significance of serum IL-6 with classic tumor markers - carcinoembryonic antigen (CEA) and squamous cell cancer antigen (SCC-Ag) - in EC patients in relation to its histological types - squamous cell carcinoma of esophagus (ESCC) and adenocarcinoma (AD) of esophagus. The study included 53 EC patients and 90 healthy subjects. Serum IL-6 and CEA levels were determined using immunoenzyme assays, while SCC-Ag - chemiluminescent assay. The diagnostic criteria and prognostic values for markers were defined. The levels of all proteins tested in EC, ESCC, and AD were higher than in healthy subjects. The percentage of elevated results was substantially higher for IL-6 (86%) than for CEA (30%) and SCC-Ag (24%) in EC, similarly as in ESCC (87%, 23%, and 33%) and AD (87%, 39%, and 13%, respectively) patients. Concentrations of IL-6 depended on distant metastases and patients' survival in EC and were significantly higher in ESCC patients with more advanced tumor stage and nodal metastases. The IL-6 area under receiver operating characteristic curve (0.92) was larger than for CEA (0.84) and SCC-Ag (0.62) in EC, likewise in ESCC (0.92, 0.87, 0.77) and AD (0.91, 0.79, 0.57, respectively). Our findings indicate better usefulness of IL-6 than classic tumor markers in the diagnosis of EC, especially in patients with ESCC.

Relationship between CDT and disease activity in rheumatoid arthritis.

The aim of this study was to estimate the serum concentration of carbohydrate-deficient transferrin (CDT) in patients with rheumatoid arthritis (RA) and the relationship between the CDT level and disease activity in RA patients. Studies were carried out in 47 female patients with RA and 32 healthy women. Disease activity of RA was evaluated using the 28-joint count Disease Activity Score (DAS 28). Serum CDT was determined by particle-enhanced immunononephelometry using the N Latex CDT test. Patients with RA had significantly lower serum concentrations of CDT compared with controls. The correlation study showed the significant negative relationship between CDT and DAS 28 (r = - 0.483, p = 0.011). There were no correlations between serum CDT level and patient's age, disease duration, number of tender and swollen joints, and degree of disability evaluated by the Health Assessment Questionnaire. The level of CDT in patients with RA was significantly decreased and confirms the changes in transferrin glycosylation which are dependent on the disease activity. Therefore, measurement of CDT in the sera of patients with RA can be useful for the evaluation of disease activity in these patients.

RANTES in exhaled breath condensate of allergic asthma patients with exercise-induced bronchoconstriction.

BACKGROUND: The response of asthmatics to exercise differs from that of healthy subjects, and the mechanisms responsible for exercise-induced bronchoconstriction (EIB) remain to be elucidated. OBJECTIVES: The aim of this study was to evaluate changes in RANTES levels in exhaled breath condensate (EBC) following intensive exercise in allergic asthmatics. METHODS: The study was conducted in a group of 19 asthmatics (11 with EIB and 8 without EIB) and 7 healthy volunteers. Changes in the concentrations of RANTES in EBC induced during the 24 h after intensive exercise were determined. Moreover, these measurements were tested for possible correlations with the results of other tests commonly associated with asthma as well as with changes in airway inflammation after exercise. RESULTS: In contrast to asthmatic patients without EIB and healthy controls, in asthmatics with EIB RANTES concentrations were statistically significantly increased in EBC collected during the first 24 h after an exercise test. There was a statistically significant correlation between the maximum increase in RANTES concentrations in EBC after exercise and either baseline exhaled nitric oxide (F(ENO)) or bronchial hyperreactivity to histamine and an increase in serum eosinophil cationic protein or F(ENO) 24 h after exercise in the EIB asthmatics. CONCLUSIONS: The increase in RANTES in asthmatic airways, promoting the migration and activation of inflammatory cells including eosinophils, may play an important role in the upregulation of airway inflammation after EIB in asthmatic patients.

Changes in RANTES and beta-thromboglobulin after intensive exercise in patients with allergic asthma.

BACKGROUND: There is increasing evidence that exercise-induced bronchoconstriction (EIB) is associated with eosinophilic airway inflammation. In the pathogenesis of EIB the role of chemokines - responsible for promoting the migration and activation of inflammatory cells - as well as blood platelets, a potential source of those chemokines, remains unclear. METHODS: The study was conducted in a group of 19 asthmatics (11 with EIB, 8 without EIB) and 8 healthy volunteers. Changes in the plasma concentrations of RANTES and beta-thromboglobulin (beta-TG) induced by intensive exercise were determined. Moreover, the possible correlation of these measurements with the results of other tests used in the diagnosis of asthma as well as laboratory tests commonly associated with asthma were investigated. RESULTS: A comparison of the concentrations of beta-TG in all groups studied at rest did not reveal any significant differences. In all groups studied, 30 min after exercise elevated beta-TG concentrations were observed; the most significant increase was revealed in asthmatics with EIB. The baseline concentrations of RANTES before exercise in both groups of asthmatics were significantly higher in comparison to the group of healthy volunteers. After exercise, in the group of patients with EIB, a significant increase in RANTES concentrations was observed. These changes correlated with an increase in other markers of airway inflammation 24 h after exercise. CONCLUSIONS: We suggest that platelet activation, resulting in elevated RANTES release, could be one of the factors responsible for the increase of airway inflammation observed in consequence of EIB in asthmatics.

Analysis of -675 4 g/5 G SERPINE1 and C-159T CD14 polymorphisms in house dust mite-allergic asthma patients.

BACKGROUND: Experimental studies indicate that endogenous plasminogen activator inhibitor-1 (PAI-1, encoded by the gene SERPINE1) modulates the immune response to lipopolysaccharide (LPS). On the other hand, LPS induces PAI-1 secretion. Activation of individual cells by LPS is facilitated by CD14. The single nucleotide polymorphisms -675 4G/5G in SERPINE1 and C-159T in CD14 are major determinants of PAI-1 and CD14 expression, respectively. OBJECTIVE: To evaluate the frequency of the -675 4G/5G SERPINE1 and C-159T CD14 polymorphisms in house dust mite (HDM) allergic asthma patients. METHODS: The polymorphisms were evaluated in unrelated inhabitants of northeastern Poland, including 372 HDM-allergic asthmatic patients and 160 healthy nonatopic control subjects using polymerase chain reaction. RESULTS: Both the C allele of CD14 and the 4G allele of SERPINE1 were more frequently encountered in HDM-allergic asthmatic patients than in healthy control individuals. When the 5G/5G-TT/CT genotype was considered as a nonrisk genotype, all other genotypes were associated with asthma. The odds ratios ranged from 3.96 (95% confidence interval, 1.56-10.1) for the 5G/5G-CC genotype to 10.7 (95% confidence interval, 5.1-24.9) for the 4G/4G-CC genotype. Bronchial reactivity to histamine and total serum immunoglobulin (Ig) E levels were predominantly associated with the 4G/5G SERPINE1 variants, while bronchial reactivity to Dermatophagoides pteronyssinus and serum concentrations of specific IgE against D pteronyssinus were predominantly associated with the C/T CD14 variants. Patients with 4G/4G-CC genotype had the lowest forced expiratory volume in 1 second and the highest bronchial reactivity. CONCLUSION: The SERPINE1 and CD14 polymorphisms studied here are associated with different aspects of bronchial reactivity and IgE response. Our results indicate that PAl-1 and CD14 may interact to affect susceptibility to allergic asthma.

Could neutrophil-gelatinase-associated lipocalin and cystatin C predict the development of contrast-induced nephropathy after percutaneous coronary interventions in patients with stable angina and normal serum creatinine values?

The value of neutrophil-gelatinase-associated lipocalin (NGAL) was highlighted as a novel biomarker for the detection of acute renal failure. We tested the hypothesis whether NGAL could represent an early biomarker of contrast-induced nephropathy (CIN) in 100 patients with normal serum creatinine values undergoing percutaneous coronary interventions (PCI). In addition, we assessed serum and urinary NGAL in relation to cystatin C, estimated glomerular filtration rate, and serum and urinary creatinine in these patients. We measured urinary and serum NGAL values before and 2, 4, 8, 24, and 48 h after the PCI. We found a significant rise in serum NGAL levels 2, 4, and 8 h after the PCI and in urinary NGAL values 4, 8, and 24 h after a PCI procedure. Cystatin C rose significantly 24 h after the procedure. The prevalence of CIN was 11%. The NGAL levels were significantly higher 2 h after the PCI (serum NGAL) or 4 h after the PCI (urinary NGAL), whereas the cystatin C values were higher only 8 and 24 h after a PCI procedure in patients with CIN. In multivariate analysis, only serum creatinine was a predictor of serum NGAL before a PCI. NGAL may represent a sensitive early biomarker of renal impairment after PCI. Serum creatinine level, the presence of diabetes, and the duration of the PCI may affect serum NGAL values and kidney function following a PCI procedure.

Alcohol dehydrogenase (ADH) isoenzymes and aldehyde dehydrogenase (ALDH) activity in the sera of patients with colorectal cancer.

Various isoenzymes of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) exist in human colorectal mucosa. In our last experiments we have shown that ADH and ALDH are present also in colorectal cancer cells. Moreover the activities of total ADH and class I isoenzymes were significantly higher in cancer tissue than healthy mucosa. This may suggest that these changes may be reflected by enzyme activity in the serum. Therefore, we have measured the activity of total ADH, and classes I-IV of this enzyme and ALDH in the sera of patients suffering from this cancer. Total ADH activity was measured by a photometric method with p-nitrosodimethylaniline (NDMA) as a substrate and ALDH activity by the fluorometric method with 6-methoxy-2-naphtaldehyde as a substrate. For the measurement of the activity of class I and II isoenzymes we employed fluorometric methods, with class-specific fluorogenic substrates. The activity of class III ADH was measured by the photometric method with formaldehyde and class IV with m-nitrobenzaldehyde as a substrate. Serum samples were taken for routine biochemical investigations from 52 patients with colorectal carcinoma before treatment. A statistically significant increase of class I ADH isoenzymes was found. Therefore the total ADH activity was also significantly increased. The total ALDH and the activity of other tested ADH isoenzymes were unchanged. We also observed the increasing tendency of ADH I activity in accordance with the advance of disease. The activity of class I ADH isoenzymes was elevated in the serum of patients with colorectal cancer. This activity was derived from colorectal cancer cells and probably from severely damaged liver by metastatic disease.

Basal serum tryptase level correlates with severity of hymenoptera sting and age.

BACKGROUND: Increased serum tryptase has been linked to the severity of the reaction after Hymenoptera stings. The aim of the study was to measure basal tryptase levels in patients with Hymenoptera venom allergy and investigate the possible correlation between these levels and the severity of sting reaction. METHODS: One hundred nine patients were included in the study. Sixty-three were wasp venom-allergic and 46 were honey bee venom-allergic. Basal serum tryptase levels were measured by UniCAP. RESULTS: Basal serum tryptase levels were elevated in 12 (11%) of the 109 patients. Levels were 5.14 pg/L (3.62-5.84), 5.3 microg/L (2.94-6.54), 5.18 microg/L (3.71-6.25), and 6.98 microg/L (4.78-12.6), for patients with sting reactions of grade I, II, III and IV (as classified by Mueller), respectively. Basal serum tryptase levels correlated significantly with the sting reaction severity (r = 0.2752; P = .004) and with age (r = 0.2906; P = .002). Sting reaction severity also correlated with age (r = 0.3654; P = .001). CONCLUSIONS: Basal serum tryptase levels were found to be elevated in 11% of venom allergic patients and correlated significantly with both sting reaction severity and age.

The activity of class I, II, III and IV alcohol dehydrogenase isoenzymes and aldehyde dehydrogenase in breast cancer.

Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) play a significant role in the metabolism of many biological substances. ADH participates in the metabolism of ethanol, retinoic acid, lipid peroxidation products, leukotriene and glutathione metabolism. ALDH is responsible for oxidation of acetaldehyde and other aldehydes and metabolism of histamine and retinoic acid. The aim of this study was to compare the metabolism in breast cancer cells and normal breast parenchyma by measuring ADH isoenzymes and ALDH activities in these tissues. Total ADH activity was measured by a photometric method with p-nitrosodimethylaniline (NDMA) as a substrate. For the measurement of the activity of ALDH and class I and II isoenzymes of ADH we employed the fluorometric methods, with class-specific fluorogenic substrates. The activity of class III alcohol dehydrogenase was detected by the photometric method with n-octanol and class IV with m-nitrobenzaldehyde as substrates. The samples were taken surgically during resection of breast carcinoma from 75 women. The activity of the class I ADH isoenzyme was significantly lower in breast cancer cells than in healthy tissues. The other tested classes of ADH had a tendency for higher levels of activity in cancer cells than in normal mammary tissue. The activity of total ADH and ALDH was also not significantly lower in the cancer cells. The decrease of activity of class I ADH isoenzyme in breast cancer tissues may be a factor of some disorders in metabolic pathways with participation of these isoenzymes that can lead to carcinogenesis.

Comparison of exhaled nitric oxide measurement with conventional tests in steroid-naive asthma patients.

BACKGROUND: Nitric oxide (NO) is a molecule with potent biological activity that plays an important role in the physiology of the respiratory system. Increased expression of inducible nitric oxide synthase (iNOS) and elevated fractional concentration of exhaled nitric oxide (F(ENO)) are seen in asthmatic patients. Measurement of F(ENO) has become increasingly recognized for use in the evaluation of bronchial inflammation during monitoring of antiinflammatory treatment. OBJECTIVES: The aim of this study was to evaluate F(ENO) in a group of steroid-naive asthmatics and assess the relationship of this parameter with the results of other tests used in the diagnosis of asthma and monitoring of antiinflammatory treatment in asthmatic patients. METHODS: The study was conducted in a group of 101 steroid-naive asthmatics (56 allergic and 45 nonallergic) and 39 healthy volunteers. All patients underwent measurement of F(ENO), skin prick tests with common inhaled allergens, analysis of serum eosinophil cationic protein (ECP) and blood eosinophilia, and flow-volume spirometry. When the forced expiratory volume in the first second (FEV1) was less than 80% of predicted, reversibility of airway obstruction with a beta2-agonist was assessed. A nonspecific bronchial provocation test with histamine was carried out in asthmatic patients with a baseline FEV1 of more than 70% of predicted. RESULTS: Compared to the healthy volunteers, F(ENO) was elevated in both groups of asthmatics. F(ENO) in the allergic asthma group was higher than in the group of nonallergic asthmatics. In allergic and nonallergic asthmatics, F(ENO) was significantly correlated with bronchial hyperresponsiveness to histamine, reversibility of airway obstruction, serum ECP levels, and blood eosinophilia. F(ENO) did not correlate with baseline FEV, in either group of asthmatics. In 31% of nonallergic and 9% of allergic patients, F(ENO) was less than 20 parts per billion. CONCLUSIONS: We suggest that measurement of F(ENO) could be clinically useful in steroid-naive asthmatics and should be more widely used in clinical practice. Measurement of F(ENO) is a noninvasive, simple, and reproducible procedure, the results of which correlate with other routinely used methods in the diagnosis of asthma. However, it is worth noting that some patients, especially those with nonallergic asthma, do not display elevated F(ENO).

An increase of collagen biosynthesis precedes other symptoms of ethanol-induced liver damage in rats.

Treatment of rats with 10% ethanol for 6 months induced a 3-4-fold increase of [3H]proline incorporation into liver collagen and enhanced the level of prolyl hydroxylase activity in liver and serum of intoxicated rats. Several diagnostic tests used to assess liver disease failed to demonstrate any symptom of liver damage. Therefore, increased collagen biosynthesis appears to precede other symptoms of liver damage induced by ethanol.

Activity of class I and II isoenzymes of alcohol dehydrogenase measured by a fluorometric method in the sera of patients with obstructive jaundice.

Using new fluorogenic substrates, we measured the activity of class I and II alcohol dehydrogenase (ADH) isoenzymes in the sera of patients with obstructive jaundice. The activity of class I isoenzymes was elevated two-fold, whereas that of class II isoenzymes was unchanged. This increase of class I isoenzymes explains low increase of total serum ADH activity. ADH isoenzymes and total enzyme activities correlated better with aminotransferases, than with alkaline phosphatase and gamma-glutamyltransferase, but we can conclude that class I ADH isoenzymes measured with fluorogenic substrates are indicative of obstructive jaundice.

Serum activities of classes I and II alcohol dehydrogenases in toxic liver damage.

The activities of classes I and II alcohol dehydrogenase isoenzymes were determined in the sera of patients with toxic hepatitis using class-specific fluorogenic substrates. The activities of total alcohol dehydrogenase and enzymes indicative of liver damage were also measured. We found a statistically significant increase of class I alcohol dehydrogenase isoenzymes. The increase in class I (two-fold) was similar to the increase of alkaline phosphatase. In a correlated study, we observed a good correlation of the activity of class II isoenzymes with alanine aminotransferase. The total alcohol dehydrogenase activity was enhanced and correlated with lactate dehydrogenase. These results demonstrated that the alcohol dehydrogenase and class I isoenzymes are indicatory enzymes of liver cell damage, and may be diagnostically useful in toxic hepatitis.

Colony stimulating factor (CSF) activity in human and murine sera.

Substances with CSF activity were studied in human and murine sera. Gel filtration of the sera on Sephadex G-200 showed that fractions with molecular weight of 14,000 and 6000 D were active in the CSF test. When mice were stimulated with endotoxin, CSF activity of both fractions was twice as high as in control animals, and CSF activity was found also in a protein peak with the molecular weight of 300,000 D. The fractions with CSF activity were sensitive to pepsin.

Tissue inhibitors of granulocyte-macrophage (GM) colony formation.

The effect of mice tissue homogenates on the inhibition of granulocyte-macrophage colony formation in vitro was studied. The potent inhibitory activity was displayed by homogenate from liver. The lowest values were found in the kidney and heart. An interrelationship between the rate of inhibition and protein concentration in the homogenates was noted. Biological role and the type of inhibitory activity are briefly discussed.

Effect of sera from acute lymphoblastic leukemia children on proliferation and differentiation of HL-60 promyelocytic leukemia cell line.

Influence of sera of children with ALL on character (dispersed or compact) and composition (granulocyte or macrophage) of colonies, formed from the human promyelocytic leukemia cell line HL-60, was estimated. An increased activities stimulating formation of dispersed G and M colonies was found in the sera of patients before therapy. Dynamics of colonies formation and their composition in the course of treatment point to some changes in the serum the levels of these activities with a tendency to full normalization in remission.

The in vivo effect of colony stimulating factors (CSFs) on phagocytic functions of granulocytes.

The effect of commercially available colony stimulating factors and mouse post-endotoxin serum on bactericidal activity and phagocytosis of bacteria by blood neutrophils was tested on mice in vivo after intraperitoneal injection. It was found that tested substances are able to induce phagocytosis and bactericidal activity of blood granulocytes. The similar patterns of these actions during 48 hours were observed for mouse IL-3 and human GM-CSF and for GCT-CM and post-endotoxin serum.

Granulocyte-macrophage-colony stimulating factor in patients with colorectal cancer.

Granulocyte-macrophage-colony stimulating factor (GM-CSF) belongs to the group of glycoproteins called colony-stimulating factors (CSFs). It has been shown that the activity of CSFs is not limited to the hematopoietic cells but can also affect the proliferation of colon carcinoma cell lines. The purpose of this investigation was to compare the serum level of GM-CSF in colorectal cancer patients to a control group, to assess the level of GM-CSF in relation to the level of carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA 19-9), and to define the sensitivity, the specificity and the predictive values of GM-CSF in colorectal cancer. In this study, the serum level of tumour markers was measured in 30 patients with colorectal cancer and in 20 healthy subjects. GM-CSF was assayed using ELISA system, CEA and CA 19-9 were measured by MEIA. The serum levels of CEA, CA 19-9 and GM-CSF were higher in the patients with colorectal cancer than in the control group. The sensitivities of CEA (63%) and CA 19-9 (56%) were lower than the GM-CSF sensitivity (80%). The specificities of tumour markers were 70% (CEA, GM-CSF) and 75% for CA 19-9. The GM-CSF predictive v values were higher than the CEA and CA 19-9 values. These results suggest that GM-CSF may be useful as tumour marker in colorectal cancer, but further studies are needed.