Enzyme Immunometric Assay Research Articles

Role of early serum beta human chorionic gonadotropin measurement in predicting multiple pregnancy and pregnancy wastage in an in vitro ET fertilization cycle

OBJECTIVE:This study was performed to assess the prognostic value of serum beta human chorionic gonadotropin (βhCG), measured on day 14 post embryo transfer (ET) for predicting multiple gestation and pregnancy wastage in women undergoing in vitro fertilization ET (IVF-ET).MATERIALS AND METHODS:This retrospective study was performed between May 2009 and November 2012. Out of the 181 women who conceived, 168 were included and the remaining 13 were excluded as their pregnancy was biochemical. Serum βhCG was measured using a chemiluminescent enzyme immunometric assay. The predictive values of serum βhCG for establishing multiple pregnancy and pregnancy wastages were calculated by receiver operating characteristic (ROC) curve analysis. Median values of serum βhCG and outcome of all pregnancies were compared.RESULTS:Out of the 168 patients who conceived after IVF treatment, 114 (68%) were viable pregnancies (delivered/ongoing). Among the viable pregnancies, 97 (85%) had a successful pregnancy outcome and the remaining 17 patients are ongoing pregnancies. Median values of βhCG (625 IU/L) among viable pregnancies was significantly (P < 0.05) higher than that of nonviable pregnancies (174 IU/L). The median values of βhCG for singleton (502 IU/L), twins (1093 IU/L), and triplets (2160 IU/L) was statistically significant (P < 0.05). Using ROC curve it was predicted that for a value of βhCG at 375 IU/L, the sensitivity of viable pregnancy was 65% and specificity of viable pregnancy was also 65%, with positive and negative predictive values of 65 and 68%, respectively. Similarly for multiple pregnancy and pregnancy wastage the predictive values of βhCG were 808 and 375 IU/L, respectively; while the sensitivity and specificity is more than 65% each.CONCLUSION:βhCG cutoff values determined on day 14 post ET by ROC curve analysis are useful in discriminating between multiple pregnancy and pregnancy losses. The cutoff value might aid in the prognosis, clinical management, and counseling of the IVF patients.

Serum level of HE4 is closely associated with pulmonary adenocarcinoma progression

The human epididymis 4 (HE4) protein is expressed in the epididymis and respiratory tract. We previously reported that HE4 is also expressed in pulmonary adenocarcinoma. The purpose of this study was to investigate serum levels of HE4 as a biological marker in pulmonary adenocarcinoma. As the trained set, 102 patients with pulmonary adenocarcinoma who underwent surgery in our institute from 2008 to 2011 were evaluated. They were compared with 58 healthy controls and 16 cases of benign lung disease. In the validation, we used 104 patients with pulmonary adenocarcinoma operated on between 2000 and 2007. Postoperative changes of serum HE4 levels were investigated in 35 patients. The level of HE4 was determined by enzyme immunometric assay and compared with clinicopathological factors. In the trained set, HE4 levels in sera in pulmonary adenocarcinoma were significantly higher than in healthy controls and benign lung disease. Receiver operating characteristic curve showed that HE4 was a good discriminator of pulmonary adenocarcinoma (cut-off point, 50.3 pM; area under curve, 0.825; 95 % confidence interval, 0.76-0.89, p < 0.001). In the validation set, serum HE4 levels were significantly correlated with age, nodal status, and carcinoembryonic antigen. Furthermore, postoperative increase of HE4 serum levels showed a significant correlation with recurrence (p = 0.032). The 5-year overall survival rate was 52.6 % in the HE4-positive group compared with 97.1 % in the HE4-negative group (p = 0.001). These data showed that HE4 expression in sera is associated with progression of pulmonary adenocarcinoma and a possible biomarker.

Decreased leptin concentration in neonates is associated with enhanced postnatal growth during the first year

Leptin regulates maternal metabolism and fetal growth by reducing food intake and increasing energy expenditure, particularly during the third trimester. In this study, we investigated the relationships between leptin and growth, and explored the longitudinal change of leptin in early postnatal life. A total of 58 infants were categorized according to gestational length and birth weight. Arterial blood samples were taken within 24 hours (Day 1), and on Days 4 and 7 of life. Plasma leptin levels were measured by commercial human leptin enzyme immunometric assay. The average serum leptin level declined in the first week of life. There was a positive correlation between leptin level and body weight on Day 4. Neonates with leptin decrease between Day 1 and Day 4 had better weight gain at one year old, and the hospital stay day was shorter. Furthermore, the full feeding days and the duration of feeding priming and full feeding days in the leptin decrease group were less than in the leptin increase group. Serum leptin was significantly decreased and positively correlated with neonates' body weight gain in the first week of life. A rapid decline in serum leptin after birth is associated with greater future weight gain and physiological advantage for infants' life.

The Relationship between Apelin and Parathyroid Hormone in Hemodialysis Patients

Both apelin and parathyroid hormone (PTH) are endogenous ligands for G-protein-coupled receptors. Apelin acts as a mitogenic agent for osteoblasts, and metabolic bone abnormalities are frequently seen in hemodialysis (HD) patients because of hyperparathyroidism. The aim of this study was to analyze plasma apelin levels in HD patients and to determine whether they are related to PTH concentrations. A total of 23 HD patients [15 men and 8 women, with a mean (SD) age of 54.2 (4.4) years and a mean body mass index (BMI) of 25.0 (4.1) kg/m2] were studied and compared with 15 healthy subjects [6 men and 9 women, with a mean (SD) age of 51.3 (13.6) years and a BMI of 27.0 (4.3) kg/m2]. Plasma apelin-36 was measured using an enzyme immunometric assay method and PTH was measured by ELISA. There was no significant difference in apelin levels between the patients [0.80 (0.6) ng/mL] and the healthy subjects [0.83 (0.23) ng/mL]. There was a positive correlation between apelin and PTH (r = 0.66, p = 0.0001). The patients with PTH >300 pg/mL had significantly higher plasma apelin levels [1.17 (0.7) ng/mL] compared with the patients with PTH <300 pg/mL [0.50 (0.15) ng/mL] (p = 0.003). In conclusion, HD patients with secondary hyperparathyroidism have high plasma apelin levels, which suggest that apelin may protect bone in HD patients by acting as an osteoblastic factor.

Apelin in hemodialysis patients: is there relationship with oxidative and inflammation markers?

Inflammation and oxidative stress are importante features associated with pathogenesis of cardiovascular disease in hemodialysis (HD) patients. Apelin is a bioactive peptide involved in a variety of physiological functions that has been associated with inflammation, however; little is known about apelin in chronic kidney disease (CKD). Thus, the purpose of this study was to analyse apelin plasma levels in HD patients and verify if there is any relationship with inflammation and oxidative markers. Twenty-four HD patients (53.6±14.4 years of age, 14 men and body mass index (BMI) of 25.0±4.2 kg/m 2 ) were studied and compared to 15 healthy subjects (51.3±13.5 years of age, 7 men and BMI of 26.3±3.7 kg/m 2 ). Plasma apelin-12 and -36 were measured using the enzyme immunometric assay (EIA) method. Plasma electronegative low density lipoprotein (LDL-) levels were measured using ELISA method. The levels of tumor-necrosis factor-α (TNF-α), interleukin-6, leptin and plasminogen activator inhibitor-1 were measured by a multiplex assay kit and C-reactive protein (CRP) by immunoturbidimetry. There was no difference between apelin-36 levels in HD patients (0.82±0.60 ng/mL) and healthy subjects (0.83±0.23 ng/mL). In contrast, apelin-12 levels were significantly higher in patients, 0.34 ±0.15 ng/mL than in healthy subjects, 0.24± 0.13ng/mL. Inflammation (TNF-α and CRP) and oxidative markers levels (LDL-) were higher in HD patients (5.4±1.3 pg/mL, 0.15 (0.33) mg/dL and 0.19±0.13U/L, respectively); however, there was not correlation among apelin-12 or -36 and inflammatory or oxidative markers. In conclusion, plasma apelin seems to be not associated with inflammatory and oxidative status in HD patients.

Long-term sertraline treatment increases expression and decreases phosphorylation of glycogen synthase kinase-3B in platelets of patients with late-life major depression

BackgroundAbnormal regulation of glycogen synthase kinase 3-beta (GSK3B) activity has been implicated in the pathophysiology of mood disorders. Many pharmacological agents, including antidepressants, can modulate GSK3B. The aim of the present study was to investigate the effect of short-and long-term sertraline treatment on the expression and phosphorylation of GSK3B in platelets of patients with late-life major depression. MethodsThirty-nine unmedicated elderly adults with major depressive disorder (MDD) were initially included in this study. The comparison group comprised 18 age-matched, healthy individuals. The expression of total and Ser-9 phosphorylated GSK3B (pGSK3B) was determined by Enzyme Immunometric Assay (EIA) in platelets of patients and controls at baseline, and after 3 and 12 months of sertraline treatments for patients only. During this period, patients were continuously treated with therapeutic doses of sertraline. GSK3B activity was indirectly estimated by calculating the proportion of inactive (phosphorylated) forms (pGSK3B) in relation to the total expression of the enzyme (i.e., GSK3B ratio). ResultsDepressed patients had significantly higher levels of pGSK3B as compared to controls (p < 0.001). Within the MDD group, after 3 months of sertraline treatment no significant changes were observed in GSK3B expression and phosphorylation state, as compared to baseline levels. However, after 12 months of treatment we found a significant increase in the expression of total GSK3B (p = 0.05), in the absence of any significant changes in pGSK3B (p = 0.12), leading to a significant reduction in GSK3B ratio (p = 0.001). ConclusionsOur findings indicate that GSK3B expression was upregulated by the continuous treatment with sertraline, along with an increment in the proportion of active forms of the enzyme. This is compatible with an increase in overall GSK3B activity, which may have been induced by the long-term treatment of late-life depression with sertraline.

A Change of Osteocalcin (OC) and Tartrate Resistant Acid Phosphatase 5b (TRACP-5b) with the Menstrual Cycle

Bone metabolism markers associated with 4 menstrual cycle phases were evaluated in 14 healthy young females without menstrual disorder. Menstrual cycle phases were confirmed with basal body temperature for 3 months, luteinizing hormone kits, and sexual hormone concentrations of serum. The bone metabolism markers used were osteocalcin (OC), which was measured by immunoradiometric assay (IRMA), and tartrate resistant acid phosphatase 5b (TRACP-5b), which was measured by enzyme immunometric assay (EIA). The highest values of OC and TRACP-5b were observed in the ovulation phase, and TRACP-5b increased significantly when compared with levels in the menstrual phase (p<0.05). Furthermore, the changes in sex-hormone secretion involved in OC and TRACP-5b showed specific patterns during the menstrual cycle. In other words, TRACP-5b levels are influenced by sex hormones produced during the menstrual period and are based on the bone-formation status. Therefore, it is presumed that the TRACP-5b levels during ovulation play a central role in bone formation and bone metabolism.

Apelin: A Peptide Involved in Cardiovascular Risk in Hemodialysis Patients?

Inflammation, oxidative stress, and obesity are important features associated with pathogenesis of cardiovascular disease, a major contributor to the mortality of hemodialysis (HD) patients. Apelin is an adipokine involved in a variety of physiological functions; however, little is known about apelin in chronic kidney disease (CKD). Thus, the purpose of this study was to analyze apelin plasma levels in HD patients and verify whether there is any relationship with inflammation, oxidative markers, and obesity. Twenty-four HD patients [53.6 ± 14.4 years, 14 men, and body mass index (BMI) of 25.0 ± 4.2 kg/m2] were studied and compared with 15 healthy subjects (51.3 ± 13.5 years, 7 men, and BMI of 26.3 ± 3.7 kg/m2). Plasma apelin-12 and -36 were measured using the enzyme immunometric assay method. Plasma electronegative low-density lipoprotein [LDL(–)] levels were measured using ELISA method, and tumor necrosis factor-α, interleukin-6, leptin, and plasminogen activator inhibitor-1 levels were measured by a multiplex assay kit. C-Reactive protein (CRP) was determined by immunoturbidimetry. Anthropometric data were also evaluated. There was no difference between apelin-36 levels in HD patients (0.82 ± 0.60 ng/mL) and healthy subjects (0.83 ± 0.23 ng/mL). In contrast, apelin-12 levels were significantly higher in patients (0.34 ± 0.15 ng/mL vs. 0.24 ± 0.13 ng/mL in healthy subjects). TNF-α, CRP, and LDL(–) levels were higher in patients; however, there was no correlation among apelin-12 or -36 and inflammatory or oxidative markers. The adiposity parameters were also not associated with apelin-12 or -36. In conclusion, plasma apelin seems to be not associated with cardiovascular risk in HD patients.

Histological evaluation of rat larynx in experimental polycystic ovary syndrome model

This study aimed at studying the histopathological effects of hyperandrogenemia and estrogen deficiency on larynx mucosa in experimentally designed polycystic ovary syndrome of female rats. Two groups of experimental polycystic ovary syndrome model were composed in healthy female rats by per oral letrozole administration of for 21 and 42 days. Also a control group which only took vehicle (saline) for 42 days was designed. Laryngeal mucosa and ovaries of all animals were examined histopathologically by light microscopy and the serum hormone levels were analyzed using a solid-phase, two-site chemiluminescent enzyme immunometric assay. Statistically significant edema, vascular engorgement, inflammation, cilia loss and differentiation of goblet cell distribution were observed when the control group and study groups were compared (p < 0.01). In serum hormonal analysis there was a significant increase in levels of androgens and decrease in levels of estrogens. In addition, polycystic appearance of ovaries in letrozole-administered groups and normal appearance of ovaries in control group have been proven histopathologically. Polycystic ovary syndrome which causes estrogen deficiency and hyperandrogenemia in fertile ages resulted in histopathological changes in laryngeal mucosa.

Assessment of ghrelin and leptin receptor levels in postmenopausal women who received oral or transdermal menopausal hormonal therapy

In postmenopausal women, an increased leptin concentration and reduced levels of ghrelin and adiponectin were observed. The aim of this study was to evaluate the concentrations of the active form of ghrelin, total ghrelin, leptin receptor, lipoprotein(a) (Lp(a)), and plasminogen activator inhibitor type 1 (PAI-1) in postmenopausal women who received oral or transdermal menopausal hormonal therapy (MHT). The study involved 76 healthy women: 46 women aged from 44 to 58 years who received oral (26) or transdermal (20) MHT; the control group consisted of 30 women aged from 44 to 54 years who did not receive MHT. The plasma concentrations of total ghrelin, the active form of ghrelin, Lp(a), and PAI-1:Ag were measured by enzyme-linked immunosorbent assay (ELISA). The concentration of the leptin receptor was measured by enzyme immunometric assay (EIA). We observed a significantly higher concentration of total ghrelin and the active form of ghrelin in women who received transdermal MHT in comparison with those who took oral MHT. We also found a significantly lower concentration of total ghrelin in women who received oral MHT compared with the control group. A higher concentration of PAI-1:Ag was found in the group of women who took transdermal MHT in comparison with those who took oral MHT and with the control group. The differences were statistically significant. Additionally, we found a significant negative correlation between the concentrations of total ghrelin and PAI-1:Ag and a positive correlation between the concentrations of total ghrelin and leptin receptor in women who received transdermal MHT. The study showed that women who used transdermal MHT had higher levels of total ghrelin than women who took oral MHT. This indicates a beneficial effect of the transdermal route of MHT. However, transdermal therapy was associated with adverse effects with regard to the observed higher levels of PAI-1:Ag, which in turn, can lead to a reduction in fibrinolytic activity.

Estimation and comparison of osteopontin levels in plasma in subjects with healthy periodontium and generalized chronic periodontitis and its assessment after scaling and root planing.

Background:Osteopontin (OPN) is a bone matrix derivative, whose levels reflect active lesions of aggravated periodontal disease accompanied by alveolar bone resorption. OPN is also a component of human atherosclerotic plaque, suggesting a role of OPN in cardiovascular diseases. The present study was conducted to assess and compare plasma OPN levels in subjects with healthy periodontium and generalized chronic periodontitis and to evaluate the effect of scaling and root planing on Plasma OPN levels of generalized chronic periodontitis subjects.Materials and Methods:40 gender matched subjects were divided into two equal groups, Group I- Healthy and Group II- Generalized chronic periodontitis, based on the Periodontal Disease Index. Blood samples were collected from the subjects at the time of clinical examination (Group I, II) and two months after Scaling and Root planning of Group II. Plasma OPN level was determined using a OPN Enzyme Immunometric Assay Kit (Quantikine).Results:The mean value of plasma OPN levels in subjects with generalized chronic periodontitis was higher (153.08 ng/ml) as compared to the subjects with Healthy periodontium (55.09 ng/ml). After treatment of generalized chronic periodontitis group, the level of plasma OPN decreased to 91.53 ng/ml.Conclusion:The findings from the study suggest that Plasma OPN levels were highest in plasma from sites with periodontal destruction; however, scaling and root planing resulted in the reduction of OPN levels.

Insulin-like growth factor 1 reference intervals for a chemiluminescent enzyme immunometric assay (Immulite 2000®) in an adult Mediterranean population

Insulin-like growth factor 1 reference intervals for a chemiluminescent enzyme immunometric assay (Immulite 2000®) in an adult Mediterranean population

Progranulin overexpression predicts overall survival in patients with glioblastoma

Despite multimodal treatment, patients with astrocytoma still face a poor survival, and identification of valuable prognostic factors is crucial to yield effective individual therapy strategies. The aim of this study was to investigate progranulin (PGRN) expression in astrocytomas and explore its association with tumor grade and overall patient survival by scoring the PGRN immunoreactivity of both tumor cells and blood vessels. About 210 astrocytoma samples with different WHO grades and 14 normal brain tissues were studied by immunohistochemistry for PGRN. Semi-quantitative RT-PCR and Western blot were carried out to confirm its expression in 35 tumor specimens. Serum levels of PGRN in glioblastoma were examined by enzyme immunometric assay. PGRN expression was almost undetectable in the normal brain tissues by immunohistochemistry but increased in both astrocytoma cells and tumor blood vessels with pathological grading. Sera in glioblastoma were significantly higher than in healthy control. In grade II astrocytoma, strong vascular PGRN expression was closely related to tumor recurrence. In glioblastoma, high total PGRN expression, strong vascular PGRN expression, and strong tumor cellular PGRN expression all correlated with decreased patient survival in univariate analysis. However, only total PGRN expression as well as vascular PGRN expression status was independently associated with patient's survival in the multivariate analysis. These results suggest that PGRN, involved in astrocytoma progression, may serve as a prognostic biomarker for glioblastoma.

Activity-dependent neurotrophic factor-derived peptide prevents alcohol-induced apoptosis, in part, through Bcl2 and c-Jun N-terminal kinase signaling pathways in fetal brain of C57BL/6 mouse

Fetal alcohol exposure is known to induce alteration in fetal brain development. In this study, we focused on neuroprotection against the effects of alcohol exposure using ADNF-9, a peptide derived from activity-dependent neurotrophic factor. We used a mouse model of fetal alcohol exposure to identify the intracellular mechanisms underlying the neuroprotective effects of ADNF-9. On embryonic day 7 (E7), weight-matched pregnant females were assigned to the following groups: (1) ethanol liquid diet (ALC) of 25% (4.49%, v/v) ethanol-derived calories; (2) pair-fed control (PF); (3) ALC combined with administration (i.p.) of ADNF-9 (ALC/ADNF-9); and (4) pair-fed combined with administration (i.p.) of ADNF-9 (PF/ADNF-9). On E13, fetal brains were collected, weighed, and apoptosis was determined using TdT-mediated dUTP nick-end labeling (TUNEL) assay. Bcl2 protein and phospho-c-Jun N-terminal kinase (JNK) levels were determined using Western blot and enzyme immunometric assay, respectively. ADNF-9 administration significantly prevented alcohol-induced reductions in fetal brain weight. In addition, ADNF-9 prevented an alcohol-induced increase in cell death in the primordium of the cerebral cortex and ganglionic eminence. Western blot analysis of the mitochondrial protein fractions revealed that ADNF-9 administration prevented an alcohol-induced reduction in the Bcl2 level. Moreover, an analysis of the proteins in the upstream signaling pathway revealed that ADNF-9 downregulated the phosphorylation of JNK. These data indicate that the mitochondrial Bcl2 pathway and JNK upstream signaling pathway are the intracellular targets of ADNF-9. The neuroprotective mechanism of action of ADNF-9 provides a direction for potential therapeutics against alcohol-induced neural damage involving mitochondrial dysfunction.

Pitfalls in the determination of human acylated ghrelin plasma concentrations using a double antibody enzyme immunometric assay

ObjectivesTo investigate the effect of hemolysis and protease inhibitors on acylated ghrelin (AG) concentrations measured with a double antibody enzyme immunometric assay that uses an acetylcholinesterase (AChE)-Fab′ conjugate. Design and methodsSamples were hemolysed or treated with PHMB (p-hydroxymercuribenzoate), PMSF (phenylmethanesulfonylfluoride) or AEBSF (4-(2-Aminoethyl) benzenesulfonyl fluoride) to prevent AG degradation. ResultsHemolysis decreased AG concentrations. PHMB or PMSF did not affect the assay. The standard curve was abolished by AEBSF but rescued by addition of a washing step prior to the AChE-Fab′ conjugate. ConclusionsHemolysis and AEBSF may affect AG determination.

Serum Adiponectin and Ghrelin, Metabolic Syndrome and Diabetes Status in Cuban Americans

Purpose: Metabolic syndrome (MetS) is associated with the development of cardiovascular disease (CVD) and type 2 diabetes. Decreases in circulating adiponectin and ghrelin have been associated with MetS. Our primary aim was to evaluate the relationship of MetS with adiponectin and ghrelin for Cuban Americans with and without type 2 diabetes. Methods: Cross-sectional study of 367 adults, self identified as Cuban extraction and randomly recruited from a mailing list of Broward and Miami-Dade counties. Fasted whole blood for adiponectin (ADPN) was collected using K3EDTA tubes and measured by ELISA. Ghrelin was assayed with fasted blood plasma by Enzyme Immunometric Assay. MetS and 10-year risk for coronary heart disease (CHD) were determined using the ATP III criteria. Results: Adiponectin (F=51.8, R2=0.21 p&lt;0.001) and ghrelin (F=12.77, R2=0.06, p&lt;0.001) differed by diabetes status (ANOVA) not age and gender. In stepwise linear regression models triglyceride levels &ge;150 mg/dL negatively corresponded (coefficient = -0.23) with ghrelin levels for persons without diabetes (F=7.45, R2=0.053, p=0.007); abdominal obesity and fasting plasma glucose predicted high sensitivity C-reactive protein (hs-CRP) for persons with and without diabetes (F=16.3, R2 = .144, p &lt;0.001). Conclusion: Low ghrelin levels were associated with MetS regardless of diabetes status. High adiponectin levels were related to a low probability for those without diabetes only. There was a positive association of hs-CRP with BMI, MetS and number of MetS components. Keywords: Metabolic syndrome, Type 2 diabetes, Cuban, Adiponectin, Ghrelin, High-sensitivity C-reactive protein

The Role of HE4 in Ovarian Cancer: Inhibiting Tumour Cell Proliferation and Metastasis

As a promising biomarker, human epididymis protein 4 (HE4) has been widely used for the early detection and differential diagnosis of ovarian cancer. This study evaluated the function of HE4 in the carcinogenesis and progression of ovarian cancer. An enzyme immunometric assay, used to detect HE4 in the serum of ovarian cancer patients, showed that the protein could discriminate between malignant and benign ovarian tumours with high specificity. An exogenous HE4 gene was transfected into ovarian cancer cell lines and an immortalized ovarian epithelial cell line. Compared with the controls, HE4 overexpression significantly promoted cell apoptosis and adhesion. Overexpression of HE4 also led to significant inhibition of cell proliferation, migration and invasiveness in vitro, as well as xenograft tumour formation in vivo. This is the first report to demonstrate the functional importance of HE4 in multiple cellular processes and indicates that HE4 may play a protective role in the progression of ovarian cancer.

DNA polymorphisms and tissue cyclooxygenase‐2 expression in oral lichen planus: a case‐control study

Oral lichen planus (OLP) is a chronic inflammatory disorder defined as a precancerous condition. Special attention has been paid to the expression of cyclooxygenase-2 (COX-2) and its potential role in development of oral squamous cell carcinoma. The identification of single nucleotide polymorphisms that affect gene function or expression and contribute to disease predisposition has become a major area of investigation toward understanding the mechanisms for cancer. The objective of this study is to investigate the association between the COX-2 765G>C gene polymorphism, tissue COX-2 expression and the development of OLP as a chronic inflammatory condition. This study was done on 50 patients with OLP and 50 healthy controls. COX-2 activity was assessed by measuring tissue prostaglandin E (PGE)2 levels by enzyme immunometric assay kit. COX-2 765G>C gene polymorphism was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR) followed by restricted fragment length polymorphism (RFLP). OLP patients showed statistically significant higher mean PGE2 than the control group. We did not observe any statistically significant differences in genotype distribution or allele frequency between the patients and the control group (P > 0.05). Odds ratio showed no statistically significant association between COX-2 765G>C polymorphism and lichen planus. The present evidence thus indicates that variation in the COX-2 gene is unlikely to be of relevance to the aetiology of OLP. As this is the first report concerning the COX-2 -765G>C gene polymorphism and the risk of OLP, additional studies with larger sample size will be required to confirm these findings.

Butyrate suppresses mRNA increase of osteopontin and cyclooxygenase-2 in human colon tumor tissue

The short chain fatty acid (SCFA) butyrate, a product of fermentation of dietary fiber in the human colon, is found to exert multiple regulatory processes in colon carcinogenesis. The aim of this study was to find out whether butyrate affects the tumor-promoting genes osteopontin (OPN) and cyclooxygenase (COX)-2, their respective proteins and/or their functional activity in matched normal, adenoma and tumor colon tissues obtained from 20 individuals at colon cancer surgery. Quantitative real-time polymerase chain reaction experiments showed increased levels of OPN and COX-2 messenger RNA in tumor tissues when compared with the adjacent normal samples (P < 0.001). The addition of butyrate reduced OPN and COX-2 mRNA expression in all tissue types compared with the related medium controls (tumor: P < 0.05). In tumor samples, a downregulation of up to median 35% (COX-2) and 50% (OPN) was observed, respectively. Thereby, tumors with lower levels of OPN basal expression were more sensitive to inhibition and vice versa for COX-2 in normal tissue. At the protein and enzyme level, which were determined by using western blot and enzyme immunometric assays, the impact of the SCFA was not clearly visible anymore. The active proteins of OPN and COX-2 (determined by prostaglandin E(2)) were found to correlate with their respective mRNA expression only in 50-63% of analyzed donors. For the first time, our data reveal new insights into the chemoprotective potential of butyrate by showing the suppression of OPN and COX-2 mRNA in primary human colon tissue with the strongest effects observed in tumors.

Platelet GSK3B activity in patients with late-life depression: Marker of depressive episode severity and cognitive impairment?

Objective. Increased GSK3B activity has been reported as a state marker of major affective episodes in patients with depression and bipolar disorder. No study so far has addressed GSK3B activity in late-life depression. The aims of the present study were to determine GSK3B activity in platelets of elderly patients with major depression, and the association between GSK3B activity and the severity of depressive symptoms and cognitive impairment. Methods. Forty drug-free elderly patients with major depressive episode were compared to healthy older adults (n = 13). Severity of the depressive episode and current cognitive state were determined by the Hamilton Depression Scale (HAM-D) and the Cambridge Cognitive Test (CAMCOG), respectively. Total- and ser-9-phosphorylated GSK3B (tGSK3B and pGSK3B) were determined in platelets by enzyme immunometric assays (EIA). GSK3B activity was indirectly inferred by the GSK3B ratio (i.e. pGSK3B/tGSK3B). Results. Elderly depressed patients had significantly lower pGSK3B levels (P = 0.03) and GSK3B ratio (P = 0.03), indicating higher GSK3B activity. Higher GSK3B activity were observed in patients with severe depressive episode (HAM-D scores >22, P = 0.03) and with cognitive impairment (CAMCOG scores <86, P = 0.01). Conclusion. The present findings provide additional evidence of the involvement of GSK3B in the pathophysiology of late-life major depression. Higher GSK3B activity may be more relevant in those patients with more severe depressive symptoms and cognitive impairment.