Ioannis Drygiannakis

Chronic venous disease progression and modification of predisposing factors

AIM This study evaluated long-term characteristics of chronic venous disease (CVD) progression and its correlation with the modification of specific risk factors. METHODS The contralateral limb of 73 patients (95% women; mean age, 48 +/- 12 years) undergoing varicose vein surgery was prospectively evaluated using physical and color duplex examination and classified by CEAP. After 5 years of follow-up, development of new sites of reflux among the contralateral, preoperatively asymptomatic limbs and modification of predisposing factors, including prolonged orthostatism, obesity, estrogen therapy (ET), multiparity, and elastic stockings use (ESU), were assessed. Data were analyzed with Pearson chi(2), t test, binary logistic regression, and Spearman rho. RESULTS Forty-eight new sites of reflux (superficial system, 37; perforators, 5; deep veins, 6) were revealed in 38 limbs (52%). CEAP scores significantly deteriorated: clinical, 2.2 +/- 0.5 from 0.1 +/- 0.03 (P < .01); anatomic, 3.8 +/- 1.2 from 2.6 +/- 2.5 (P < .05); disability, 1.9 +/- 0.7 from 0 (P < .01); and severity, 7.9 +/- 2.4 from 2.7 +/- 2.2 (P < .01). Patient compliance to predisposing factor modification was low; no change was observed during follow-up (orthostatism, P = .9; obesity, P = 0.7; ET, P = .9; multiparity, P = .4; ESU, P = .3). CVD progression was significantly lower in patients who controlled orthostatism vs those who maintained orthostatism or initiated it (P < .001) and in patients who controlled preoperative obesity vs those who became obese or maintained obesity (P < .001). Non-ESU patients had a significantly higher incidence of CVD progression vs those who started ESU or continued during the study (P < .001). By binary logistic regression analysis, orthostatism (P = .002; B coefficient value [BCV] = 1.745), obesity (P = .009; BCV = 1.602), and ESU (P = .037; BCV = 0.947) were independent predictive factors for CVD progression, whereas multiparity (P = .174) and ET (P = .429) were not. CONCLUSIONS In about half of patients with unilateral varicosities, CVD developed in the contralateral initially asymptomatic limb in 5 years. CVD progression consisted of reflux development and clinical deterioration of the affected limbs. Obesity, orthostatism, and noncompliance with ESU were independent risk factors for CVD progression, but ET and multiparity were not. Maintenance of a normal body weight, limitation of prolonged orthostatism, and systematic ESU may be recommended in patients with CVD to limit future disease progression.

Proinflammatory cytokines induce crosstalk between colonic epithelial cells and subepithelial myofibroblasts: Implication in intestinal fibrosis

BACKGROUND AND AIMS Colonic epithelial cells and adjacent subepithelial myofibroblasts are important counterparts in the pathogenesis of intestinal inflammation and fibrosis. We investigated the possible crosstalk between them, whilst focusing on the mucosal inflammation pathways that potentially trigger intestinal fibrosis. METHODS We studied the effects of proinflammatory cytokines (IL-1α, TNF-α, IFN-γ) on human colonic epithelial cell lines and the effects of epithelial cell-conditioned media on primary human colonic subepithelial myofibroblasts isolated from normal controls or patients with inflammatory Crohns disease along with the corresponding 18CO cell line. Readouts included production of TGF-β and TIMP-1, total collagen synthesis, matrix metalloproteinases MMP-2 and MMP-9 and myofibroblast migration/mobility. RESULTS Proinflammatory cytokines upregulated TGF-β and TIMP-1 in colonic epithelial cells. Conditioned medium from these epithelial cell cultures induced production of MMP-9 and collagen and inhibited the migration/mobility of subepithelial myofibroblasts. MMP-9 production depended on endothelin receptor A signalling on responding myofibroblasts. Collagen up-regulation was independent of TGF-β, CTGF, TF and endothelin. Subepithelial myofibroblasts isolated from Crohns disease patients had similar responses to those isolated from normal controls, with the exception of higher basal collagen production. CONCLUSIONS Our study indicates that colonic epithelial cells may respond to an inflammatory milieu by inducing myofibroblast functions similar to those observed during intestinal fibrosis.

Increased expression of chemokine receptor CCR3 and its ligands in ulcerative colitis: the role of colonic epithelial cells in in vitro studies

Human colonic epithelial cells express T helper type 1 (Th1)‐associated chemoattractants, yet little is known about the production of Th2‐associated chemoattractants. CCL11/eotaxin‐1, CCL24/eotaxin‐2 and CCL26/eotaxin‐3 are known to attract CCR3‐expressing, Th2‐polarized lymphocytes. We studied constitutive and inflammation‐induced expression and production of CCR3 together with its ligands in the colon and peripheral blood of patients with inflammatory bowel disease (IBD) by flow cytometry, reverse transcription–polymerase chain reaction (RT–PCR) and enzyme‐linked immunosorbent assay (ELISA). We further defined the regulated expression of these chemokines by RT–PCR and ELISA using cultured human epithelial cell lines. A higher fraction of peripheral T lymphocytes were found to be positive for CCR3 in patients with ulcerative colitis (UC) compared to Crohns disease (CD), while almost no CCR3+ T cells were found in normal controls (NC). Similarly, higher and more frequent expression of CCR3 was observed in colonic biopsies from patients with UC, regardless of the disease activity, when compared to CD or NCs. Serum CCL11/eotaxin‐1 was increased significantly in UC (306 ± 87 pg/ml) and less so in CD (257 ± 43 pg/ml), whereas CCL24/eotaxin‐2, and CCL26/eotaxin‐3 were increased only in UC. Colonic expression of the three chemokines was minimal in NCs but high in inflammatory bowel diseases (especially UC) and was independent of disease activity. Th2, and to a lesser extent Th1, cytokines were able to induce expression and production of all three eotaxins from colonic epithelial cells in culture. CCR3 and ligands over‐expression would appear to be a characteristic of UC. The production of CCR3 ligands by human colonic epithelial cells suggests further that epithelium can play a role in modulating pathological T cell‐mediated mucosal inflammation.

Pancreatic Autoantibodies in Greek Patients with Inflammatory Bowel Disease

Pancreatic autoantibodies (PAbs) have been suggested as a specific but not sensitive marker for Crohns disease (CD). The aim of this study was to assess the value of detecting PAbs in Greek patients with ulcerative colitis (UC) and CD. Sera were collected from 150 patients with IBD (73 with UC and 77 with CD), 31 cases with non-IBD intestinal inflammation, 16 cases with other autoimmune diseases, and 104 healthy controls. Determination of PAbs was performed by a standard indirect immunofluorescence technique. PAbs were detected in 18 of 73 (24.7%) samples from UC patients and in 32 of 77 (41.6%) samples from CD patients. The prevalence of positive PAbs was significantly higher in CD than in UC (P= 0.04). None of the 104 samples from healthy controls and the 31 cases with non-IBD intestinal inflammation had detectable PAbs. One patient with Sjogrens syndrome was PAbs positive. No association of PAbs with IBD activity, IBD localization, or medical treatment was found. Patients with stenotic CD had a significantly higher prevalence of PAbs positivity (60%) compared with patients with inflammatory (28.6%) and fistulizing (41.2%) disease (P= 0.02). The prevalence of PAbs in Greek CD patients was found to be similar to that in previous reports. In contrast to these studies we found also increased prevalence of PAbs in UC patients. These findings suggest that PAbs should be considered as a specific marker for IBD rather than for CD.

Octreotide induces caspase activation and apoptosis in human hepatoma HepG2 cells

AIM To investigate the role of octreotide on cellular proliferation and apoptosis of human hepatoma (HepG2) cells. METHODS We studied cellular proliferation, apoptosis and the possible internal caspase-mediated apoptosis pathway involved, after treatment of HepG2 carcinoma cells with octreotide in comparison with the apoptosis caused by tumor necrosis factor-α (TNF-α). Activities of caspase-3, caspase-9, caspase-8 and caspase-2 were studied, while apoptosis was investigated through detection of DNA fragmentation and through identification of apoptotic cells with the annexin-V/propidium iodide flow cytometric method. RESULTS After an initial increase in HepG2 cellular proliferation, a significant inhibition was observed with 10⁻⁸ mol/L octreotide, while TNF-α dose-dependently decreased proliferation. Early and late apoptosis was significantly increased with both substances. Octreotide significantly increased caspase-3, caspase-8 and caspase-2 activity. TNF-α significantly increased only caspase-2. Cellular proliferation was decreased after treatment with octreotide or TNF-α alone but, in contrast to TNF-α, octreotide decreased proliferation only at concentrations of 10⁻⁸ mol/L, while lower concentrations increased proliferation. CONCLUSION Our findings are suggestive of caspase-mediated signaling pathways of octreotide antitumor activity in HepG2 cells, and indicate that measurements of serum octreotide levels may be important, at least in clinical trials, to verify optimal therapeutic drug concentrations.

Ciprofloxacin decreases survival in HT-29 cells via the induction of TGF-β1 secretion and enhances the anti-proliferative effect of 5-fluorouracil

Background and purpose:  Fluoroquinolones are potent anti‐microbial agents with multiple effects on host cells and tissues. Previous studies have highlighted their pro‐apoptotic effect on human cancer cells and an immunoregulatory role in animal models of inflammatory bowel disease. We examined the effect of ciprofloxacin on proliferation, cell cycle and apoptosis of HT‐29 cells, a human colonic epithelial cell line sensitive to transforming growth factor (TGF)‐β1‐mediated growth inhibition and its role in TGF‐β1 production. We also examined the effect of ciprofloxacin on proliferation of HT‐29 cells in combination with 5‐fluorouracil (5‐FU), a well‐established pro‐apoptotic agent.

A concentration-dependent effect of ursodeoxycholate on apoptosis and caspases activities of HepG2 hepatocellular carcinoma cells

Clinical observations suggest that ursodeoxycholate (UDCA) may protect from hepatocellular carcinoma in cirrhotic patients. Increased apoptosis of malignant cells is a candidate mechanism. Decreased apoptosis of cholangiocytes is proposed as a mechanism for the favourable effect of UDCA in primary biliary cirrhosis. We therefore studied the effects of different concentrations of UDCA on HepG2 cell proliferation, apoptosis and caspases activities. Apoptotic features and activities of the effector or initiator caspases-8, -9, -3 and -2 after treatment of HepG2 cells with different concentrations of UDCA alone or in combination with TNF-alpha were examined. Apoptosis was detected by DNA fragmentation and flow cytometric determination of apoptotic cells with Annexin-V/PI. UDCA significantly inhibits cell proliferation only at high concentrations, but increases apoptosis at low concentrations and protects from apoptosis at higher concentrations. TNF-alpha induced DNA fragmentation is potentiated by UDCA, but flow cytometry indicates protection from early apoptosis and increase in cell survival by low and intermediate UDCA concentrations. UDCA differentially activates initiator and effector caspases in different concentrations. These data demonstrate that the effect of UDCA on caspase activation and apoptosis of HepG2 cells is concentration-dependent and activation of the caspase cascade is not always translated into increased apoptosis. Serum levels of UDCA should be possibly monitored and dosage of the drug adjusted according to the required effect.

Low prevalence of liver-kidney microsomal autoantibodies of type 1 (LKM1) in hepatitis C seropositive subjects on Crete, Greece.

BackgroundHepatitis C is a serious problem on the Greek island of Crete, where a high prevalence of antibodies against hepatitis C (anti-HCV) has recently been reported. This article reports the findings of a study carried out in Crete, which investigated the prevalence of serum autoantibodies in patients with chronic hepatitis C.Patients and MethodsOne hundred and forty two patients (59 men and 83 women), who were found anti-HCV seropositive in two hospitals and two Primary Health Care Centres in Crete, were eligible. Sixty healthy blood donors (46 men, 14 women), which were negative to anti-HCV, were used as the control group. They were randomly selected from those attending Rethymnon Hospital. Autoantibodies were identified using the indirect immunofluorescence (IFL) technique on human epithelial cells from larynx cancer (HEp-2 cells), rat liver-kidney-stomach substrate (CT3) and Chrithidia Luciliae (CL).ResultsSerum autoantibodies were detected in 104 HCV patients, yielding an overall prevalence of 73.2%. The most frequent autoantibodies were antinuclear antibodies (ANA), positive in 72 patients (50.7%). Anti-smooth muscle antibodies (ASMA) were detected in 33 patients (23.2%). Only one patient was positive for LKM1 autoantibodies. No autoantibodies were found in 38 patients (26.7%). Autoantibodies were also found in 5 out of the 60 examined healthy blood donors (8.3%).ConclusionsAutoantibodies, mainly ANA and ASMA are very common in HCV seropositive patients from Crete. By contrast LKM1 autoantibodies are exceptionally rare in these patients.

A giant adrenal lipoma presenting in a woman with chronic mild postprandial abdominal pain: a case report.

IntroductionAdrenal lipomas are rare, small, benign, non-functioning tumors, which must be histopathologically differentiated from other tumors such as myelolipomas or liposarcomas. They are usually identified incidentally during autopsy, imaging, or laparotomy. Occasionally, they may present acutely due to complications such as abdominal pain from retroperitoneal bleeding, or systemic symptoms of infection. We report a giant adrenal lipoma (to the best of our knowledge, the second largest in the literature) clinically presenting with chronic mild postprandial pain.Case presentationA 54-year-old Caucasian woman presented several times over a period of 10 years to various emergency departments complaining of long-term mild postprandial abdominal pain. Although clinical examinations were unrevealing, an abdominal computed tomography scan performed at her most recent presentation led to the identification of a large lipoma of the left adrenal gland, which occupied most of the retroperitoneal space. Myelolipoma was ruled out due to the absence of megakaryocytes, immature leukocytes, or erythrocytes. Liposarcoma was ruled out due to the absence of lipoblasts. The size of the lipoma (16 × 14 × 7 cm) is, to the best of our knowledge, the second largest reported to date. After surgical resection, our patient was relieved of her symptoms and remains healthy six years postoperatively.ConclusionPhysicians should be aware that differential diagnosis of mild chronic abdominal pain in patients presenting in emergency rooms may include large adrenal lipomas. When initial diagnostic investigation is not revealing, out-patient specialist evaluation should be planned to enable appropriate further investigations.

Expression of a splice variant of CXCR3 in Crohn's disease patients; indication for a lymphocyte—epithelial cell interaction

Background and Aim:  T‐lymphocyte migration is implicated in the pathogenesis of Crohns disease (CD) and ulcerative colitis (UC). CXC chemokines MIG, IP‐10, and I‐TAC act by binding to CXCR3 receptor on T‐lymphocytes. We investigated the role of these chemokines and their receptor in patients with UC, CD, and normal controls (NC).