Prostaglandins are key regulators of ion transport in the kidney. In MDCK cells, which model distal tubule cells, the transcription of the Na,K-ATPase <em>beta</em>1 subunit is regulated by PGE1 and PGE2. To identify the <em>EP</em> receptors that mediate transcriptional regulation, transient transfection studies are conducted using the human <em>beta</em>1promoter/luciferase construct, pH<em>beta</em>1-1141 Luc. The involvement of <em>EP</em>1 and <em>EP</em>2 receptors is indicated by studies with the <em>EP</em>1 selective agonist 17-phenyl trinor PGE2, and the <em>EP</em>2 selective agonist butaprost (which stimulate), as well as by studies with the antagonists SC-51089 (<em>EP</em>1 specific) and AH 6809 (<em>EP</em>1 and <em>EP</em>2 specific). Consistent with the involvement of Gs coupled <em>EP</em>2 receptors, is that the PGE1 stimulation is inhibited by the PKAI expression vector (encoding the protein kinase A (PKA) inhibitory protein), as well as by the myristolated PKA inhibitory peptide PKI. In addition to this evidence (for the involvement of <em>EP</em>2 receptors), evidence for the involvement of <em>EP</em>1 receptors in the PGE1 mediated stimulation of Na,K-ATPase <em>beta</em> subunit gene transcription includes the stimulatory effect of 17-phenyl trinor PGE2, as well as the inhibitory effects of SC-51089. Also consistent with the involvement of Gq coupled <em>EP</em>1 receptors, the PGE1 stimulation is inhibited by the PKCI vector (encoding the PKC inhibitory domain), the PKC inhibitor Go 6976, thapsigargin, as well as the calmodulin antagonists W7 and W13.

Despite the widespread use of TVS for diagnosing EP and extensive literature on the subject, there is no consensus regarding the best positivity criterion for adnexal findings or the performance characteristics of TVS. We conducted a literature search to identify original studies presenting suitable data on the use of TVS for the diagnosis of EP. The data were combined to determine the sensitivity and specificity of four sonographic criteria for EP, listed in order from most to least stringent: Criterion A, living extrauterine pregnancy; criterion B, extrauterine gestational sac containing yolk sac or embryo; criterion C, empty "tubal ring" or extrauterine gestational sac containing yolk sac or embryo; and criterion D, any adnexal mass other than a simple cyst. Positive and negative predictive values were computed using Bayes' theorem. Ten studies involving a total of 2216 patients, 565 with EP and 1651 without EP, were included in our analysis. Based on the combined data from these studies, criteria A, B, and C all have high specificities (99.5-100%) and positive predictive values (97.8-100%) but low sensitivities (20.1-64.6%) and mediocre negative predictive values (78.5-89.1%). Criterion D, the most lax criterion, has the most uniformly excellent characteristics, with only slightly lower specificity (98.9%) and positive predictive value (96.3%) but considerably higher sensitivity (84.4%) and negative predictive value (94.8%). The performance characteristics of TVS criteria for EP, computed by pooling data from published studies, indicate that the appropriate TVS criterion to diagnose EP is any noncystic adnexal mass. These performance characteristics can be used as a basis for comparing TVS with other proposed diagnostic modalities for EP.

A strain of Bacillus licheniformis 8-37-0-1 with high exopolysaccharide (EPS) production ability was isolated and identified based on morphological and physiological characteristics and phylogenetic analysis of 16S rDNA sequences. A new type of EPS was isolated from the strain fermentation broth by enzymolysis, isopropanol precipitation, anion-exchange, and gel-filtration chromatography. The new EPS was determined as homogeneous, with a molecular weight of 2.826 x 10(4), as determined by High-Performance Size-Exclusion Chromatography Multi-Angle Laser Light Scattering analysis. Its structural characteristics were investigated and elucidated by methylation analysis, partial acid hydrolysis, gas-liquid chromatography mass spectrometry, Fourier transform infrared, and nuclear magnetic resonance spectroscopy. Based on obtained data, the EPS was found to be a levan containing a (2-->6)-linked backbone with a single beta-d-fructose at the C-1 position every seven residue, on average, along the main chain. Preliminary in vitro tests revealed that EPS could significantly stimulate the proliferation of spleen lymphocyte.

Contagious bovine pleuropneumonia is a severe respiratory disease of cattle that is caused by a bacterium of the Mycoplasma genus, namely Mycoplasma mycoides subsp. mycoides (Mmm). In the absence of classical virulence determinants, the pathogenicity of Mmm is thought to rely on intrinsic metabolic functions and specific components of the outer cell surface. One of these latter, the capsular polysaccharide galactan has been notably demonstrated to play a role in Mmm persistence and dissemination. The free exopolysaccharides (EPS), also produced by Mmm and shown to circulate in the blood stream of infected cattle, have received little attention so far. Indeed, their characterization has been hindered by the presence of polysaccharide contaminants in the complex mycoplasma culture medium. In this study, we developed a method to produce large quantities of EPS by transfer of mycoplasma cells from their complex broth to a chemically defined medium and subsequent purification. NMR analyses revealed that the purified, free EPS had an identical β(1->6)-galactofuranosyl structure to that of capsular galactan. We then analyzed intraclonal Mmm variants that produce opaque/translucent colonies on agar. First, we demonstrated that colony opacity was related to the production of a capsule, as observed by electron microscopy. We then compared the EPS extracts and showed that the non-capsulated, translucent colony variants produced higher amounts of free EPS than the capsulated, opaque colony variants. This phenotypic variation was associated with an antigenic variation of a specific glucose phosphotransferase permease. Finally, we conducted in silico analyses of candidate polysaccharide biosynthetic pathways in order to decipher the potential link between glucose phosphotransferase permease activity and attachment/release of galactan. The co-existence of variants producing alternative forms of galactan (capsular versus free extracellular galactan) and associated with an antigenic switch constitutes a finely tuned mechanism that may be involved in virulence.

The aim of the present study was to evaluate in a mouse model of spinal cord injury (SCI) the effect of the treatment with ethyl pyruvate (EP). Spinal cord injury was induced by the application of vascular clips (force of 24 g) to the dura via a four-level T5-T8 laminectomy in mice. Treatment with EP (75, 25, or 8.5 mg/kg) 1 and 6 h after the SCI significantly decreased (a) the degree of spinal cord inflammation and tissue injury (histological score), (b) neutrophil infiltration (myeloperoxidase activity), (c) nitrotyrosine formation and iNOS expression, (d) proinflammatory cytokines expression, (e) nuclear factor kappaB activation, (f) extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase phosphorylation, and (g) apoptosis (TUNEL staining, Fas ligand, Bax, and Bcl-2 expression). Moreover, EP (75, 25, or 8.5 mg/kg) significantly ameliorated in a dose-dependent manner the loss of limb function (evaluated by motor recovery score). Taken together, our results demonstrate that EP treatment reduces the development of inflammation and tissue injury associated with spinal cord trauma.

Cyclooxygenase 2 and release of prostaglandin E2 are involved in many responses including inflammation and are upregulated during cellular senescence. However, little is known about the role of lipid inflammatory mediators in senescence. Here, we investigated the mechanism by which the COX-2/PGE2 axis induces senescence. Using the NS398 specific inhibitor of COX-2, we provide evidence that reactive oxygen species by-produced by the COX-2 enzymatic activity are negligible in front of the total senescence-associated oxidative stress. We therefore investigated the role of PGE2 by invalidating the PGE2 synthases downstream of COX-2, or the specific PGE2 receptors, or by applying PGE2 or specific agonists or antagonists. We evaluated the effect on senescence by evaluating the senescence-associated proliferation arrest, the percentage of senescence-associated beta-galactosidase-positive cells, and the expression of senescent molecular markers such as IL-6 and MCP1. We show that PGE2 acting on its EP specific receptors is able to induce both the onset of senescence and the maintenance of the phenotype. It did so only when the PGE2/lactate transporter activity was enhanced, indicating that PGE2 acts on senescence more via the pool of intracellular EP receptors than via those localized at the cell surface. Treatment with agonists, antagonists and silencing of the EP receptors by siRNA revealed that EPEPEPs intracrine pathway.

OBJECTIVE

In osteoarthritis (OA), bradykinin (BK) is known to contribute to pain and synovitis, but not to cartilage degradation. Here, we investigated effects of BK and its antagonists on chondrocytes, cells involved in cartilage homeostasis.

METHODS

BK receptor density and affinities of BK, its analogues and antagonists were measured in cultured human and rat chondrocytes by radioligand binding. Effects of BK were assessed by accumulation of inositol phosphates (IP) and release of interleukin (IL)-6 and IL-8.

RESULTS

Density of [³H]-BK binding sites was higher (13-30-fold) and BK evoked a greater (48-fold) IP production, in human than in rat chondrocytes. The BK B₂ receptor antagonists MEN16132 and icatibant displayed similar binding affinity. MEN16132 was 40-fold more potent than icatibant in the IP assay. In human chondrocytes, BK increased release (over 24 h) of IL-6 and IL-8, effects blocked by MEN16132 but not by the B₁ receptor antagonist Lys-[Leu⁸][desArg⁹]BK. BK-induced release of IL-6, but not of IL-8, was partially inhibited by indomethacin (10 µM) and nordihydroguaiaretic acid (10 µM). Antagonists for the prostanoid EP receptors (AH6809 10 µM; L-798,196, 200 nM; L-161,982, 1 µM) were ineffective. Dexamethasone (100 nM) partially inhibited release of both IL-6 and IL-8. Inhibitors of intracellular downstream signalling pathways (SBBAY-117085, 5 µM) indicated the involvement of p38 MAPK and the activation of NF-κB.

CONCLUSIONS

BK mediated inflammatory changes and cartilage degradation and B₂ receptor blockade would, therefore, be a potential treatment for OA.

METHODS

A 7-year-old Quarter Horse gelding was hospitalized in Ocala, Fla, because of lethargy, fever, anorexia, and swelling of distal aspects of the limbs. A tentative diagnosis of equine piroplasmosis (EP) was made on the basis of examination of a blood smear. The case was reported to the Florida State Veterinarian, and infection with Babesia equi was confirmed. The subsequent investigation included quarantine and testing of potentially exposed horses for B equi and Babesia caballi infections, tick surveillance, and owner-agent interviews.

RESULTS

210 horses on 25 premises were tested for infection with EP pathogens. Twenty B equi-infected horses on 7 premises were identified; no horses tested positive for B caballi. Seven horses, including the index case, had clinical findings consistent with EP Dermacentor variabilis was considered the only potential tick vector for B equi collected, and all D variabilis specimens tested negative for Babesia organisms via PCR assay. Results of the epidemiological investigation suggested that B equi was spread by use of shared needles and possibly blood transfusions. All horses that tested positive were involved in nonsanctioned Quarter Horse racing, and management practices were thought to pose substantial risk of transmission of blood-borne pathogens.

RESULTS

Final outcome of B equi-infected horses was euthanasia, death from undetermined causes, or shipment to a US federal research facility.

CONCLUSIONS

This investigation highlights the importance of collaboration between private veterinary practitioners, state veterinary diagnostic laboratories, and regulatory officials in the recognition, containment, and eradication of foreign animal disease.

Previous studies demonstrated that the extracellular polysaccharides (EPSs) produced by Lactobacillus delbrueckii OLL1073R-1 (LDR-1) improve antiviral immunity, especially in the systemic and respiratory compartments. However, it was not studied before whether those EPSs are able to beneficially modulate intestinal antiviral immunity. In addition, LDR-1-host interaction has been evaluated mainly with immune cells while its interaction with intestinal epithelial cells (IECs) was not addressed before. In this work, we investigated the capacity of EPSs from LDR-1 to modulate the response of porcine IECs (PIE cells) to the stimulation with the Toll-like receptor (TLR)-3 agonist poly(I:C) and the role of TLR2, TLR4, and TLR negative regulators in the immunoregulatory effect. We showed that innate immune response triggered by TLR3 activation in porcine IECs was differentially modulated by EPS from LDR-1. EPSs treatment induced an increment in the expression of interferon (IFN)-α and IFN-β in PIE cells after the stimulation with poly(I:C) as well as the expression of the antiviral factors MxA and RNase L. Those effects were related to the reduced expression of A20 in EPS-treated PIE cells. EPS from LDR-1 was also able to reduce the expression of IL-6 and proinflammatory chemokines. Although further in vivo studies are needed, our results suggest that these EPSs or a yogurt fermented with LDR-1 have potential to improve intestinal innate antiviral response and protect against intestinal viruses.

Objective To perform a meta-analysis of migraine biomarkers in cerebrospinal fluid (CSF) and of corresponding blood concentrations. Methods We conducted a systematic search for studies that measured biochemical compounds in CSF of chronic or episodic migraineurs and non-headache controls. Subsequent searches retrieved studies with blood measurements of selected CSF biomarkers. If a compound was assessed in three or more studies, results were pooled in a meta-analysis with standardised mean differences (SMD) as effect measures. Results Sixty-two compounds were measured in 40 CSF studies. Most important results include: increased glutamate (five studies, SMD 2.22, 95% CI: 1.30, 3.13), calcitonin gene-related peptide (CGRP) (three studies, SMD: 3.80, 95% CI: 3.19, 4.41) and nerve growth factor (NGF) (three studies, SMD: 6.47, 95% CI: 5.55, 7.39) in chronic migraine patients and decreased β-endorphin (β-EP) in both chronic (four studies, SMD: -1.37, 95% CI: -1.80, -0.94) and interictal episodic migraine patients (three studies, SMD: -1.12, 95% CI: -1.65, -0.58). In blood, glutamate (interictal) and CGRP (chronic, interictal and ictal) were increased and β-EP (chronic, interictal and ictal) was decreased. Conclusions Glutamate, β-EP, CGRP and NGF concentrations are altered in CSF and, except for NGF, also in blood of migraineurs. Future research should focus on the pathophysiological roles of these compounds in migraine.

Plasma cell tumors are lymphoid neoplasms with an uncontrolled proliferation of B cells. These are divided into localized forms (solitary bone plasmocytoma -SBP- and extramedullary plasmocytoma -EP-) and disseminated forms (multiple myeloma-MM-). The SBP is a rare and controversial disease. The aim of this article is the analysis of this entity based on the presentation of a 64-year-old man without previous medical history, with a mass in the left mandibular angle extending to the parotid region on the same side. The panoramic radiography, computed tomography and magnetic resonance imaging showed an osteolytic lesion 6.5 x 5 x 6.7 cm in the mandibular angle with infiltration of the masticator space and left parotid region. The normality of the extension study, and histopathological examination confirmed the diagnosis of SBP. The patient received treatment with radiotherapy with good outcome.

<A<em>b</em>stractText>Platinum-<em>b</em>ased chemotherapy, consisting of etoposide and cisplatin (<em>EP</em>), has <em>b</em>een the cornerstone of therapy for extensive-stage small-cell lung cancer (ES-SCLC) for decades. Despite the marked initial sensitivity of SCLC to chemotherapy, <em>EP</em> regimens cannot avoid the emergence of drug resistance in clinical practice. With the rise of new chemotherapy regimens in recent years and the primary resistance or insensitivity of ES-SCLC to <em>EP</em> regimens, it is desira<em>b</em>le to <em>b</em>e a<em>b</em>le to identify patients with resistant or insensitive ES-SCLC.</A<em>b</em>stractText><A<em>b</em>stractText>The sequencing and drug sensitivity data of SCLC cell lines were provided <em>b</em>y The Genomics of Drug Sensitivity in Cancer Project (GDSC). The data regarding sensitivity to etoposide of 54 SCLC cell lines were analyzed, and etoposide-sensitive cell lines and etoposide-resistant cell lines were differentiated according to the IC50 values defined <em>b</em>y the GDSC. ROC curve analysis was performed on all mutations and com<em>b</em>inations of mutations to select the optimal panel to predict resistance to etoposide.</A<em>b</em>stractText><p><div>(<em>b</em>)Results</<em>b</em>)</div>ROC analysis of etoposide resistance revealed that the most significant single gene mutation indicating resistance to etoposide was <i>CSMD3</i>, and the accuracy of predicting resistance to etoposide proved to <em>b</em>e the highest when there was any mutation in <i>CSMD3/PCLO/RYR1/<em>EP</em>B41L3</i>, area under the curve =0.804 (95% confidence interval: 0.679-0.930,<i>P</i><i><0.001</i>).</p><p><div>(<em>b</em>)Conclusion</<em>b</em>)</div>This study found that a panel with four genes (<i>CSMD3, <em>EP</em>B41L3, PCLO, and RYR1</i>) can accurately predict sensitivity to etoposide. These findings provide new insights into the overall treatment for patients with ES-SCLC that is resistant or insensitive to etoposide.</p>

OBJECTIVE

Regulated expression of cell adhesion molecules could be critical in the proliferation, sequestration, and maintenance of stem/progenitor cells. Therefore, we determined fetal and adult stage-specific roles of cell adhesion in liver cell compartments.

METHODS

We performed immunostaining for the adhesion molecules, E-cadherin and Ep-CAM, associated proteins, beta-catenin and alpha-actinin, hepatobiliary markers, albumin, alpha-fetoprotein, and cytokeratin-19, and the proliferation marker, Ki-67. Expression of albumin was verified by in situ mRNA hybridization.

RESULTS

In the fetal liver, hepatoblasts showed extensive proliferation with wide expression of E-cadherin, beta-catenin, and alpha-actinin, although Ep-CAM was expressed in these cells less intensely and focally in the cell membrane to indicate weak cell adhesion. Hepatoblasts in ductal plate and bile ducts showed less proliferation and Ep-CAM was intensely expressed in these cells throughout the cell membrane, indicating strong adhesion. In some ductal plate cells, beta-catenin was additionally in the cytoplasm and nucleus, suggesting active cell signaling by adhesion molecules. In adult livers, cells were no longer proliferating and E-cadherin, beta-catenin, and alpha-actinin were expressed in hepatocytes throughout, whereas Ep-CAM was expressed in only bile duct cells. Some cells in ductal structures of the adult liver with Ep-CAM coexpressed albumin and cytokeratin-19, indicating persistence of fetal-like stem/progenitor cells.

CONCLUSIONS

Regulated expression of Ep-CAM supported proliferation in fetal hepatoblasts through weak adhesion and helped in biliary morphogenesis by promoting stronger adhesion in hepatoblasts during this process. Restriction of Ep-CAM expression to bile ducts in the adult liver presumably facilitated sequestration of stem/progenitor cells. This stage-specific and cell compartment-related regulation of adhesion molecules should be relevant for defining how liver stem/progenitor cells enter, exit, and remain in hepatic niches during both health and disease.

A recently developed ultrasound phase-locked echo-tracking system makes it possible to measure non-invasive pulsatile vessel diameter changes, and, in combination with blood-pressure measurement, to calculate pressure strain elastic modulus (Ep) and stiffness (beta). The reproducibility in measurements of pulsatile diameter changes with this system was evaluated. Also the precision of indirect blood-pressure measurements, as compared to the simultaneously measured intra-arterial blood pressure was tested. The resulting reproducibility in pressure strain elastic modulus (Ep) and stiffness (beta) was evaluated. Intra-observer variabilities in measuring pulsatile diameter changes were 16% for the abdominal aorta, 10% for the common carotid artery, and 15% for the common femoral artery, respectively. Intra-observer variabilities for Ep and beta were 21% for both in the abdominal aorta, 17% for both in the common carotid artery, and 18% for both in the common femoral artery, respectively. There were only small differences in indirect and direct measurement of systolic blood pressure, whereas indirect blood pressure measurement systematically overestimated the diastolic blood pressure, on average by 20%. The variabilities in indirect blood pressure measurements were 2% for the systolic and 3% for the diastolic blood pressure, respectively. Inter-observer variability in the investigation of the common carotid artery was 10% for the pulsatile diameter changes, and 21% and 23% for Ep and beta, respectively. Thus, the echo-tracking system represents a reliable system for estimation of pressure strain elastic modulus and stiffness. However, Ep and beta are systematically underestimated by 25-30%, when used in combination with indirect blood pressure measurements.

Extracellular polymeric substances (EPS) of biological origin are ubiquitous in excess sludges and can be applied as an underlying bioflocculant, owing to their high content of macromolecules and cations. However, low flocculating activity limits the feasibility of their practical applications. This study provides a novel EPS fractionation approach to improve their flocculability by extracting an active EPS fraction and removing the others with low flocculability. The results showed that for two excess sludges (called sludge A and sludge B), the tightly bound EPS (TB-EPS) fraction possessed a high flocculating rate to kaolin suspension compared with the other EPS fractions [i.e., supernatant, slime, and loosely bound EPS (LB-EPS) fraction] (>54.1+/-1.4% vs <7.8+/-1.6%). High bioflocculability of TB-EPS fraction could be attributable to high contents of macromolecules (330-1200 kDa) and trivalent cations (Fe(3+) and Al(3+)). Further investigation reveals that the TB-EPS fraction caused aggregation of particles by bridging and sweep flocculation.

Arterial stiffness is emerging as an important risk marker for cardiovascular disease. Ultrasound-based measurements of arterial stiffness are in use by several large epidemiological studies. The reliability of ultrasonic measurements of arterial stiffness was assessed as part of one of these, the Atherosclerosis Risk in Communities (ARIC) study. ARIC, a prospective, four-center epidemiological study, used B-mode ultrasound with an electronic tracking device to measure arterial stiffness of the carotid artery. Oscillometric blood pressure measures were obtained before and after the arterial wall tracking. Measurement variability was estimated in 36 volunteers who were scanned at three visits conducted at 7- to 14-day intervals. Between- and within-person components of variation were estimated for arterial diameter and blood-pressure measurements. The correlation (R) between repeated measurements for pulse pressure, the percent change in arterial diameter (strain), and the percent and absolute change in the arterial area were 0.69, 0.67, 0.66 and 0.81, respectively. The R for the stress-strain elastic modulus (Ep), arterial distensibility, and arterial compliance were 0.66, 0.67, and 0.77, respectively. The R for the pressure-adjusted diameter change (i.e., diameter change adjusted for diastolic and pulse pressures) was 0.75. In summary, the ultrasonic measurements of arterial stiffness employed in the ARIC study demonstrate excellent short-term repeatability, demonstrating their utility in field settings.

beta-lipotrophin (beta-LPH) and beta-endorphin (beta-EP) plasma levels were measured by radioimmunoassay after glass powder extraction and Sephadex G-75 column chromatography in plasma samples from controls (ten healthy males and twenty-six young women in early follicular phase), from eighty-two pregnant women in weeks 9-40 after their last menstrual period, from nine women just after delivery and the cord blood of their neonates, in fifteen mixed cord blood samples and in seven amniotic fluid samples obtained by amniocentesis. No sex differences were found in beta-LPH (120.6 +/- 8.5 pg/ml) or beta-EP (31.1 +/- 2.4 pg/ml) plasma levels or in their molar ratio (1.34 +/- 0.09) (MR). beta-LPH plasma levels increased in early pregnancy (13-16 weeks) (185.0 +/- 27.1 pg/ml) and remained high until weeks 21-24, then declining to levels similar to those of controls. beta-EP plasma levels were significantly depressed in weeks 9-12 (20.7 +/- 5.3 pg/ml), subsequently increasing to a maximum at weeks 36-37 (42.7 +/- 6.8 pg/ml). beta-LPH/beta-EP molar ratio was about double normal in early pregnancy and decreased to normal in the second half. The present data indicate that beta-LPH and beta-EP present different patterns throughout pregnancy and that beta-EP levels increase progressively, reaching the highest concentrations at term. At delivery, both beta-LPH and beta-EP showed maximum values (beta-LPH: 230.2 +/- 20.4 pg/ml; beta-EP: 78.0 +/- 7.4 pg/ml) and a MR of 1.02 +/- 0.10 indicating that stressful situations, such as labour, stimulate a simultaneous rise in beta-LPH and beta-EP plasma levels. Cord blood specimens showed a wide range of values (beta-LPH:75-347 pg/ml; beta-EP: 16-287 pg/ml) with a MR of 1.21 +/- 0.14. Amniotic fluid samples obtained late in the third trimester of pregnancy were characterized by beta-LPH levels of 119.4 +/- 26.4 pg/ml and beta-EP levels of 29.6 +/- 7.5 pg/ml.

OBJECTIVE

To evaluate the continuous metabolic syndrome score (cMetS) in Indian children and to investigate its relationship with the risk of carotid arterial stiffness.

METHODS

Data on weight, height, mean arterial pressure, serum lipids, insulin, glucose, carotid intima-media thickness and stiffness parameters, that is, pulse wave velocity (PWV), elasticity modulus (Ep), stiffness index (β) and arterial compliance (AC), were assessed in 236 children (6-17 years) from Pune city, India. cMetS was computed using standardized Z-scores for metabolic syndrome (MS) components. cMetS cutoff was obtained by receiver operating characteristic curve analysis across MS components.

RESULTS

cMetS was lowest (-3.6±2.0) in normal children and highest (3.3±2.4) in MS children. cMetS increased progressively with number of risk components. The cutoff of cMetS yielding maximal sensitivity (80%) and specificity (94%) for predicting the presence of MS was -0.8 (area under the curve=0.921 (95% CI: 0.877-0.964)). In children with cMetS above -0.8, average PWV (4.3±0.6 m s(-1)), β (3.8±1.2) and Ep (50.4±14.5 kPa) were significantly higher than the respective values (3.7±0.5 m s(-1); 3.4±0.8; 37.0±10.0 kPa) in children with cMetS below -0.8, whereas AC was lower (1.2±0.5 mm(2) kPa(-1)) in children with cMetS above -0.8 as against AC (1.4±0.3 mm(2) kPa(-1)) in children with cMetS below -0.8 (P<0.05), demonstrating the risk of stiffness with increasing score. Pearson's correlation coefficients of cMetS with PWV (r=0.575), β (r=0.347), AC (r=-0.267) and Ep (r=0.530) were statistically significant (P<0.01).

CONCLUSIONS

Results demonstrate the usefulness of cMetS over individual MS components as a better tool for assessment of atherosclerotic risk in children.

The relationship of endogenous opiates in patients with polycystic ovarian disease (PCOD) and their influence on body weight was studied. The study group consisted of 19 women with PCOD. They were amenorrheic, hirsute, and hyperandrogenic, and their average weight was 124% of the ideal body weight. They had luteinizing hormone/follicle-stimulating hormone ratios greater than or equal to 2. The control group consisted of ten women with regular ovulatory menses. Plasma beta-endorphin (beta-EP) was measured by using a very specific radioimmunoassay. beta-Lipotropin (beta-LPH) was entirely removed from the sample by preincubation of the plasma with rabbit anti-beta-LPH/Sepharose complex (Pharmacia, New Brunswick, NJ). The mean +/- standard deviation of the plasma beta-EP in the control group was 70.18 +/- 18.06 pg/ml, and the mean +/- standard deviation of beta-EP in the study group was 185.6 +/- 93.4 pg/ml, which was significantly higher than the control levels (P less than 0.001). A significant correlation was also found between plasma beta-EP level and the patient's weight in the PCOD group (r = 0.462, P = 0.025). The data from this study suggest that the elevated levels of endogenous opiates may be involved in the pathophysiology of PCOD and be related to inappropriate secretion of gonadotropins influencing body weight.

Our aim was to evaluate the effects of sympathetic excitation and elevation of blood pressure on mechanical properties of common carotid and femoral arteries by wave intensity analysis (WIA). The diameters and arterial stiffness parameters of right common carotid artery (RCCA) and right common femoral artery (RCFA) in healthy young men were measured by WIA at baseline and during cold pressor test (CPT). In addition, the blood pressure and heart rate were recorded simultaneously. The heart rates and blood pressures increased during CPT compared with baseline, while the pulse pressures remained unchanged. The diameters of RCCA increased slightly, while those of RCFA did not change. The Peterson's pressure modulus (Ep), augment index (AI), and pulse wave velocity from β (PWVβ) increased obviously, while arterial compliance (AC) decreased with no change in stiffness index (β) of both arteries during CPT when compared with baseline. There was an obvious increase in pulse wave velocity from wave intensity (PWV_WI) of RCCA, while the PWV_WI of RCFA showed no significant change during CPT. The sympathetic nervous system exerts a more marked tonic restraint on RCFA than on RCCA. The Ep, AC, AI, PWVβ of RCCA, and RCFA are much affected by variations in blood pressure and sympathetic status, while the β of both arteries are less vulnerable to these factors and are more reliable in reflecting the actual arterial stiffness; The PWV_WI appears to be suitable only for evaluating the stiffness of RCCA instead of RCFA.

Epithelial cell adhesion molecule (Ep-CAM) is expressed in a several epithelial tissues and carcinomas, but not on mature hepatocytes. Here, we analysed the expression of Ep-CAM in 230 patients suffering from various liver diseases like chronic hepatitis B and C (HBV and HCV infection), chronic autoimmune hepatitis (AIH), chronic alcoholic liver disease (ALD), primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), hereditary hemochromatosis and dysplastic nodules (DNs) as well as hepatocellular carcinomas (HCCs) and cholangiocellular carcinomas (CCCs) by immunohistochemistry. De novo hepatocellular Ep-CAM expression was found in 75.9% of ALD (22/29), 63.6% of HCV (21/33) and 55.6% of each AIH and HBV cases (5/9 and 15/27, respectively). Lower Ep-CAM expression levels were observed for primary sclerosing liver diseases (PBC and PSC) with 25% (3/12) and 7.7% (1/13) of cases. Moreover, only 14.3% of HCCs (9/63) manifested expression, while all CCCs showed strong Ep-CAM expression (5/5). For DNs and hereditary hemochromatosis, Ep-CAM expression was found in 10 and 50% (3/30 and 2/4), respectively. In HBV and HCV, Ep-CAM expression correlated significantly with inflammatory activity as assessed by histological parameters and to the extent of fibrosis. In addition, for HCV also transaminase levels correlated significantly with Ep-CAM expression. Our results indicate that de novo Ep-CAM expression in hepatocytes is frequent in inflammatory liver diseases and is potentially linked to regenerative activity. CCCs and Ep-CAM positive HCCs may represent an attractive target group for Ep-CAM-directed immunotherapies, yet unwanted toxicity may limit the use of such strategies due to Ep-CAM expression in biliary epithelium and several chronic liver diseases such as HBV-and HCV-hepatitis.

In mice injected with formalin into the hindpaw, the 5-HT1A receptor agonists, 8-OH-DPAT and flesinoxan, equipotently inhibited the early phase (EP) and late phase (LP) of licking. At higher doses, they provoked ataxia and inhibited the writhing elicited by intra-abdominal acetic acid. The antagonists, (-)-alprenolol, (-)-tertatolol, WAY-100,135 and S 15931 were more potent against the LP than the EP. They also inhibited writhing, and only at very high doses did they elicit ataxia. In rats, 8-OH-DPAT and flesinoxan increased the current required to elicit vocalisation upon electrical stimulation of the tail. The action of 8-OH-DPAT was blocked by WAY-100,135, which, like other antagonists, was inactive alone. Interestingly, a low dose of 8-OH-DPAT partially inhibited the antinociceptive action of the mu-opioid agonist, morphine, the action of which was dose-dependently facilitated by (-)-alprenolol and S 15931. Administered s.c., 8-OH-DPAT elicited spontaneous tail-flicks (STFs) in rats: these were abolished by WAY-100,135, (-)-tertatolol, (-)-alprenolol and S 15931. STFs were also eliminated by s.c. or i.t. administration of the alpha 2-adrenergic receptor agonist, clonidine, the GABAA agonist, muscimol or the GABAB agonist, baclofen. The mu-opioid, morphine, blocked STFs only at high doses and the kappa-opioid agonists, U 50,488 and U 69,593, even at supra-ataxic doses, were inactive. Antagonists at neurokinin (NK)1 (RP 67580), NK2 (SR 48,968) and bradykinin (BK)2 (Hoe 140) receptors, as well as aspirin, did not block STFs, though indomethacin was effective. Antagonists at the glycine B site coupled to the NMDA receptor, L 687,414, L 701,324 and (+)-HA966, blocked STFs. Furthermore, (+)-HA 966 and the competitive NMDA receptor antagonist, CPP, were active upon i.t. administration. STFs were also blocked by s.c. or i.t. administration of the AMPA antagonists, YM 900 and NBQX. In conclusion, the influence of 5-HT1A ligands upon nociception is dependent upon the algesiometric paradigm. Intriguingly, modulation of 5-HT1A receptor-mediated STFs reveals parallels to neuropathic pain.

Thermophilic microorganisms (4001-4014), described as aerobic or facultatively anaerobic, endospore forming with growth optima temperatures in the range of 60 to 80 degrees C, have been isolated from hot marine springs around Ischia and from hydrothermal vents in the gulf of Naples. Mucous colonies are been selected for the recovery of new strains producing exopolysaccharides (EPS). To induce the biosynthesis of new exopolysaccharides, different sugars were tested as carbon sources in the media. The production of EPS in the strain 4009 reached 60 mg/l using trehalose as carbon source, increasing the yield of about 1000 fold. The 4001-EPS was a mannan with a molecular weight of 380.000 D and with a complex primary structure. In fact, the analysis of the permethylated polysaccharide in GC-MS, showed the presence of mannose, glucose, galactose, mannosamine in the relative ratio of 1:0.1:tr :tr, respectively. Nuclear magnetic resonance spectrum of the exopolysaccharide confirmed the presence of a repetitive unity formed by seven monosaccharides, six with alpha gluco/galacto configuration and one residue with beta conformation.

OBJECTIVE

The aim of the study was to evaluate carotid arterial stiffness and intima media thickness (IMT) in obese children in comparison with healthy children, and to examine associations of lipid profile and blood pressure with carotid artery morphology.

METHODS

Anthropometric and blood parameters were assessed in 44 overweight, 95 obese (6-17 years) and 69 healthy age-matched normal children. Percent body fat was measured by dual-energy X-ray absorptiometry and stiffness and IMT of the common carotid artery were evaluated using Aloka alpha 10 equipment.

RESULTS

Anthropometric and lipid parameters were significantly higher in overweight and obese than in normal children (p < 0.05). Stiffness (beta), pulse wave velocity (PWV), elastic modulus (Ep) and blood pressure were significantly higher in obese and overweight children than in normal children (p < 0.05). However, overweight and obese children did not show any significant difference in IMT compared with normal children (p>> 0.1). Significant positive correlation of PWV, beta and Ep and negative correlation of arterial compliance with body fat and triglyceride was noted (p < 0.05). Different multinomial regression models for each e-Tracking parameter indicated that the relative risk of hypertension was highest with high PWV, followed by LDL cholesterol, Ep and body fat.

CONCLUSIONS

PWV may be considered an important marker for evaluation of early functional changes of the carotid artery in children and adolescents.