Elevated circulating levels of matrix metalloproteinases MMP-2 and MMP-9 in pseudoxanthoma elasticum patients

Pseudoxanthoma elasticum (PXE) is a rare disorder predominantly affecting the skin, the eyes, and the cardiovascular system. The disease is caused by mutations in the ABCC6 gene and characterized by ectopic calcification and extracellular matrix (ECM) alterations. Matrix metalloproteinases (MMPs) play a pivotal role in the process of ECM remodeling. In the present study, we investigated matrix metalloproteinases MMP-2 and MMP-9 in PXE patients compared to healthy controls. We analyzed the serum concentrations of MMP-2 and MMP-9 in a cohort of 69 German PXE patients and in 69 healthy, age-, and sex-matched control subjects using commercially available ELISA assays. We found elevated concentrations of both MMPs in the sera of PXE patients. MMP-2 levels were significantly higher in patients than controls (231 ± 5.89 vs 202 ± 5.17 ng/ml, p = 0.0002), as were MMP-9 levels (841 ± 65.9 vs 350 ± 30.8 ng/ml, p < 0.0001). Our findings point to an involvement of matrix metalloproteinases in PXE pathology. ECM remodeling in PXE is reflected by elevated levels of circulating MMP-2 and MMP-9. Those MMPs might, therefore, be applicable as serum markers for the matrix-degradative process in PXE.     

Serum Matrix Metalloproteinase-2 Levels Indicate Blood–CSF Barrier Damage in Patients with Infectious Meningitis

Objective Protein components in cerebrospinal fluid (CSF) are maintained at a specific concentration by a dynamic gradient between the capillary and intrathecal spaces via the blood–cerebrospinal fluid barrier (BCB) in the brain and spinal cord. Permeability to proteins increases when there is structural damage to the BCB. Matrix metalloproteinase-2 (MMP-2; gelatinase A) has been shown to degrade type IV collagen, a major component of the cellular basement membrane. We analyzed α2 macroglobulin (α2M) indices and evaluated the relationship between α2M, as an indicator of BCB permeability, and MMP-2, which degrades the extra-cellular matrix in patients with infectious meningitis. Materials and Methods Albumin levels in CSF or serum were determined by turbidimetric immunoassay, or bromcresol green assay, respectively. α2M levels in CSF or serum were measured with enzyme-linked immunosorbent assay, or laser-nephelometry, respectively. Serum MMP-2 levels were determined by enzyme immuno assay. We calculated the α2M index, i.e. the ratio of α2M (CSF / serum) to albumin (CSF / serum; α2M in CSF / α2M in serum × albumin in serum / albumin in CSF). Results α2M indices were significantly increased in infectious meningitis compared to healthy controls (p < 0.05). They were highest in bacterial meningitis, and there was a significant difference between viral or mycotic and bacterial meningitis (p < 0.05). Serum MMP-2 levels were increased in infectious meningitis, being highest in bacterial meningitis, where they were significantly different from healthy controls (p < 0.05). There was a significant positive correlation between serum MMP-2 levels and α2M indices (r = 0.64, p < 0.0001). Conclusion Markedly increased levels of serum MMP-2 in infectious, especially bacterial, meningitis may reflect the degree of damage to the BCB.     

Subacute Inflammatory Activation in Subjects with Acute Coronary Syndrome and Left Ventricular Dysfunction

Several lines of evidence indicate that increased inflammatory cytokine levels can be used for risk prediction in patients with acute coronary syndrome (ACS). This study therefore aimed to evaluate correlations between levels of soluble interleukin (IL)-2 receptor (sIL-2r), IL-6, and IL-8 and in-hospital incidence of acute heart failure (AHF) and left ventricular (LV) systolic dysfunction in the subacute phase of ACS. In 48 consecutive patients with ACS, circulating levels of sIL-2r, IL-6, and IL-8 were ascertained 72–96 h after onset of symptoms. Clinical data, LV function, and in-hospital incidence of AHF were also evaluated. IL-8 levels were significantly higher in patients with pulmonary edema (1,829 ± 2,496 vs 456 ± 624 pg/ml, p < 0.05); sIL-2r, IL-6, and IL-8 levels were increased proportionally to Killip class (r = 0.35, p < 0.05; r = 0.48, r = 0.47, p < 0.01) and in patients with LV ejection fraction (LVEF) < 30%. Levels of sIL-2r were inversely related to LVEF in subjects with acute myocardial infarction (r = −0.51, p < 0.05). Soluble IL-2r and IL-8 levels were related to mitral regurgitation severity (r = 0.34, p < 0.05; r = 0.37, p < 0.05). Levels of sIL-2 were proportional to LV end-diastolic diameter (r = 0.49, p < 0.001) and LV end-systolic diameter (r = 0.58, p < 0.001). Number of cytokines with circulating values above upper level of normal was significantly correlated with Killip class and LVEF (r = 0.40, r = −0.38, p < 0.05). sIL-2r, IL-6, and IL-8 are increased in patients with ACS and systolic dysfunction or AHF. These data suggest that inflammatory cytokine activity detectable in peripheral blood may be useful in identifying subjects with a worse clinical course.     

Diagnostic value of the matrix metalloproteinase-8 rapid test for detecting microbial invasion of the amniotic cavity

The purpose of this paper was to compare the MMP-8 PTD Check (MPC) test with other indirect tests for detecting microbial invasion of the amniotic cavity (MIAC). Amniotic fluid (AF) was analyzed in 155 women for white blood cell (WBC) count, glucose concentration, and an MPC test and evaluated for MIAC using cultures for aerobic/anaerobic bacteria and mycoplasmas and polymerase chain reaction (PCR) of chlamydia. The median AF glucose concentration was lower and the median AF WBC count was higher in women with MIAC than in women without MIAC (p < 0.01 and p < 0.001, respectively). Also, the positive rate of the MPC test was higher in women with MIAC than in women without MIAC (p < 0.001). The sensitivities of AF glucose concentration, AF WBC count, and the MPC test for the detection of MIAC were 58.6%, 75.9%, and 86.2%, respectively. The specificities for the detection of MIAC were 76.2%, 80.2%, and 74.6%, respectively. We conclude that the MPC test is a rapid, easily performed, and accurate indirect method for detecting MIAC. Presented at the 27th Annual Meeting of the Society for Maternal–Fetal Medicine (SMFM), San Francisco, CA, 5–10 February 2007.     

Time Course of Endotoxin-Induced Airways’ Inflammation in Healthy Subjects

Few data are available on the kinetic of the airways’ inflammation induced by inhaled endotoxin in a given subject. The purpose of this study was to evaluate in healthy subjects the time-related endotoxin-induced airways’ inflammation. The cells counts from the induced-sputum were evaluated before, 6 and 24 h, and 7 days after an exposure to 20 mcg inhaled endotoxin, in eight pre-selected volunteers. To avoid interference of the induced-sputum procedure on the response to endotoxin, each time-point was evaluated in randomized order at 2-weeks interval after three separate inhalations of endotoxin. A significant rise of the relative number of lymphocytes (p < 0.05) and polymorphonuclear neutrophils (PMN; p < 0.02) and of the absolute number of PMN (p < 0.05) occurring at 6 h, followed by an increase of the absolute number of the total viable cells (p < 0.01), macrophages (p < 0.001), neutrophils (p < 0.01), and lymphocytes (p < 0.05) at 24 h after endotoxin inhalation. The inflammatory response recovered totally after 7 days. In human beings, the inhalation of endotoxin induced a transient airway inflammation after 6 h, peaked at 24 h and recovered after 7 days. When repeated endotoxin inhalations are used as a model of inflammation, a wash-out period of at least 7 days should be applied between each exposure in each subject.     

Hepatoprotective Effect of Infliximab,an Anti-TNF-α Agent,on Carbon Tetrachloride-Induced Hepatic Fibrosis

To assess the effect of infliximab, an anti-tumor necrosis factor (TNF)-α agent, on the carbon tetrachloride (CCl₄)-induced hepatic fibrosis in rats. Rats were randomized into three groups (n = 9). The control group received only intraperitoneal (i.p.) olive oil. Hepatic fibrosis was induced by repeated i.p. injections of 1.5 ml/kg CCl₄ (1:3 mixture with olive oil) for 5 weeks in the remaining two groups which were also injected subcutaneous saline or 2 mg/kg infliximab. Infliximab reduced the levels of aspartate aminotransferase and alanine aminotransferase (p < 0.05 for both). The scores of hepatic necrosis, inflammation and fibrosis, and expression of α-smooth muscle actin were lower in the infliximab-treated group than the CCI₄-treated group (p < 0.01, p < 0.001, p < 0.01, p < 0.001, respectively). However, there was no significant difference in terms of liver tissue and plasma malondialdehyde, and serum TNF-α levels, while infliximab relatively reduced the level of transforming growth factor-β₁ (373.0 ± 153.1 vs. 280.8 ± 127.1 pg/ml). Treatment with infliximab attenuated the necro-inflammation and fibrogenesis in the CCI₄-induced hepatic fibrosis, and thus it might be effective as a therapeutic anti-fibrotic agent.     

Serum C-reactive Protein (CRP) Levels in Cancer Patients are Linked with Tumor Burden and are Reduced by Anti-hypertensive Medication

High levels of CRP relate with advanced disease and poor prognosis of cancer patients. CRP serum levels were measured in 684 cancer patients who had undergone complete surgery or inoperable patients. Patients with inoperable tumors had significantly higher CRP levels (1.21 ± 2.2 vs. 0.40 ± 0.4 mg/dL; p < 0.0001). No association with gender, diabetes, autoimmune disease, thyroid disease or allergy was noted. Significantly higher CRP levels were noted in operated patients with hypertension (0.55 ± 0.5 vs. 0.35 ± 0.4; p = 0.001), coronary disease (0.73 ± 0.8 vs. 0.39 ± 0.4; p = 0.01) and obesity (0.51 ± 0.5 vs. 0.37 ± 0.4; p = 0.04). On the contrary, analysis in the group of inoperable patients showed that hypertensive patients had significantly lower CRP levels (0.64 ± 1.0 vs. 1.36 ± 2.4; p = 0.008). Although the tumor itself is the main factor defining increased CRP levels in cancer patients, hypertension, coronary disease and obesity are also linked with high CRP levels. Anti-hypertensive drugs appear as potent suppressors of the tumor-induced CRP production.     

The Treatment with Antibody of TNF-α Reduces the Inflammation,Necrosis and Fibrosis in the Non-alcoholic Steatohepatitis Induced by Methionine- and Choline-deficient Diet

To assess the effects of anti-TNF-α antibody (infliximab) in experimental steatohepatitis induced by methionine- and choline-deficient (MCD) diet. The study included thirty rats. One group received normal rat food, and two groups received MCD diet. The treatment group received a single dose intra-peritoneal infliximab (4 mg/kg), at week 8. MCD diet increased levels of AST, ALT, TNF-α, TGF-β₁, tissue and plasma MDA (p < 0.05 for each). Moreover, it led to steatosis, ballooning degeneration, inflammation, fibrosis and increased actin expression, histopathologically (p < 0.05 for each). In this experimental steatohepatitis anti-TNF-α antibody decreased the levels of AST, ALT, TGF-β₁ and plasma and tissue MDA (p < 0.05 for each). Moreover, inflammation, necrosis, actin expression and fibrosis decreased in anti-TNF-α group compared to placebo group (p < 0.05 for each). This study indicates that anti-TNF-α antibody is effective on necrosis, inflammation and fibrosis in the experimental model of non-alcoholic steatohepatitis, induced by MCD diet.     

Does the Presence of Anti-CCP Autoantibodies and Their Serum Levels Influence the Severity and Activity in Rheumatoid Arthritis Patients?

This study aims to investigate the association of the presence and of the titer of autoantibodies against cyclic citrullinated peptides (aCCP), with clinical manifestations and disease activity in a cohort of patients with rheumatoid arthritis (RA). From January 2000 through December 2005, 135 patients were diagnosed with RA at the Rheumatology Unit of our hospital. Demographic, clinical, laboratory, and therapeutic parameters were evaluated in all patients at study entry and at every follow-up visit. Positivity in aCCP and also their levels were determined for all patients. At the end of the study, we reevaluated the above parameters, dividing patients into aCCP positive and aCCP negative. From 135 patients, 53.3% were aCCP positive. The majority of aCCP-positive patients were males (p < 0.001), positive to rheumatoid factor (p < 0.001) and current smokers (p < 0.05). At diagnosis, aCCP-positive patients presented with higher tender joint counts (p < 0.001) and swollen joint counts (p < 0.001), and exhibited more active disease, expressed by higher disease activity scores for 28 joints (DAS-28) (p < 0.001). At the end of the study, aCCP-positive patients also displayed more active disease, with higher DAS-28 (p < 0.001), and more severe disease, as this was indicated by the higher radiological Larsen score (p < 0.001). The serum levels of aCCP were not found to be associated with disease activity and severity. In early RA, the presence of aCCP is associated with increased disease activity and severity. This was found to be independent of circulating levels of aCCP.     

Increased type IIA secretory phospholipase A2 expression contributes to oxidative stress in end-stage renal disease

End-stage renal disease (ESRD) patients exhibit increased in vivo oxidative stress conceivably contributing to cardiovascular mortality. The type IIA secretory phospholipase A₂ (sPLA₂) has proatherogenic activity. We explored the hypothesis that sPLA₂ contributes to oxidative stress generation and endothelial dysfunction in ESRD patients and transgenic (tg) mice. Patients with ESRD had increased in vivo oxidative stress as assessed by plasma isoprostane levels (p < 0.001). Active sPLA₂ in plasma was substantially increased compared with healthy controls (1,156 ± 65 versus 184 ± 5 ng/dL, p < 0.001) and correlated with plasma isoprostanes (r = 0.61, p < 0.001). Correspondingly, human sPLA₂ tg mice display increased generation of reactive oxygen species within aortic vascular smooth muscle cells, leading to severe endothelial dysfunction (maximal vasodilation in response to 10 μmol/L acetylcholine, sPLA₂ 36 ± 8%, controls 80 ± 2% of phenylephrine-induced vasoconstriction). Increased vascular oxidative stress in sPLA₂ tg mice is dependent on the induction of vascular cyclooxygenase (COX)2 expression. Conversely, ESRD patients show increased formation of COX2-derived prostaglandins (p < 0.05) correlated with plasma sPLA₂ (r = 0.71, p < 0.05). Our data indicate that increased expression of sPLA₂ might represent a novel causative risk factor contributing to the increased cardiovascular disease morbidity and mortality in ESRD.     

Role of syndecan-3 polymorphisms in obesity and female hyperandrogenism

The heparan sulfate proteoglycan syndecan-3 (SDC3) is a novel regulator of feeding behavior and body weight. Recently, an association of SDC3 polymorphisms with obesity has been observed in Koreans. As female obesity is associated with hyperandrogenism and infertility, we studied the role of SDC3 polymorphisms in female individuals undergoing diagnostics prior to infertility treatment. For this purpose, endocrine parameters and body mass index of 249 women were assessed. Genotyping of V208I, D303N, and T329I was performed with TaqMan technology using lymphocyte-derived DNA and allelic discrimination polymerase chain reaction. Chi-square test, Student’s t test, and one-way analysis of variance were used for statistical analysis. We find that an infrequent genotype and allele variation of T329I correlated with obesity (p = 0.028). Genotype and alleles of V208I were associated with luteinizing hormone (p = 0.007 and p = 0.001, respectively), luteinizing hormone/follicle-stimulating hormone (p = 0.002 and p < 0.005, respectively), 17 hydroxyprogesterone (p = 0.007 and p = 0.001, respectively), androstenedione (p = 0.046 and p = 0.013, respectively), and sex hormone-binding globulin (p = 0.021). We conclude that marked ethnic differences of the SDC3 SNP distribution in our European population could account for correlations less predominant compared to Koreans. While infrequent variations of T329I correlated with obesity, V208I was associated with endocrine parameters related to hyperandrogenism. These findings indicate that SDC3 polymorphisms could contribute to the link between female hyperandrogenism and obesity and suggest a novel potential role for SDC3 as a modulator of gonadal steroid function.     

Advanced oxidation protein products in obese women: its relation to insulin resistance and resistin

Obesity is a major risk factor for insulin resistance and type 2 diabetes mellitus (T2DM). Resistin, an adipocyte-secreted hormone, is thought to take a part in the development of insulin resistance and T2DM. The aim of this study was to characterise the changes in circulating levels of resistin and proinflammatory cytokines tumour necrosis factor-alpha (TNF-α) and interleukin (IL)-6 in diabetic and prediabetic obese patients and to explore their relationship to insulin resistance. Attempts were also made to see whether resistin levels are related to the degree of oxidative stress, as determined by the measurement of advanced oxidation protein products (AOPPs). The study groups consisted of obese diabetic (BMI: 30–42 kg/m², n=28) and prediabetic (BMI: 29–41 kg/m², n=23) women. Fourteen healthy women, with BMI in the range 21.5–25.5 kg/m², were taken as controls. Serum levels of TNF-a, IL-6, resistin, glucose, insulin and AOPPs were measured. Insulin resistance was calculated by the homeostasis model assessment (HOMA-IR). Diabetic and prediabetic obese patients had increases in serum resistin and TNF-α levels (P<0.01 and P<0.001, respectively). IL-6 levels in diabetic patients were significantly higher than in prediabetics (P<0.05). AOPP levels were also significantly higher in diabetics than prediabetics and controls (P<0.05 and P<0.001, respectively); and positively correlated with blood glucose. Insulin was significantly associated with circulating resistin and TNF-α. The development of insulin resistance may contribute to the elevation of circulating resistin or vice versa. Determination of AOPPs may be helpful for monitoring the impaired glucose metabolism in obesity.     

Nadroparin Sodium Activates Nrf2/HO-1 Pathway in Acetic Acid-Induced Colitis in Rats

Effects of nadroparin sodium, a low molecular weight heparin, in colitis was investigated by analyzing proteins implicated in nuclear factor E2-related factor-2/heme oxygenase-1 (Nrf2/HO-1) and nuclear factor kappa B (NF-κB) pathways. Twenty-eight rats were used. Colitis was induced by acetic acid (AA). Nadroparin sodium was given to prevention and treatment groups in addition to AA. Colitis was assessed histologically and levels of proteins were analyzed with Western blot. Nadroparin not only prevented and ameliorated the AA-induced colitis histopathologically but also decreased expression of colon NF-κB, activator protein-1, cyclooxygenase-2, tumor necrosis factor-alpha, and IL-6, which were significantly increased in group AA compared to control. The accumulation of Nrf2 in nuclear fraction and HO-1 found low in group AA was increased with nadroparin (p < 0.05). The mean malondialdehyde level increased with AA and was decreased significantly with nadroparin prevention and treatment (p < 0.001). Nadroparin sodium has both protective and therapeutic effects against colonic inflammation via exerting anti-oxidative and anti-inflammatory effects by modulating Nrf2/HO-1 and NF-κB pathways.     

C-reactive protein and nitric oxide levels in ischemic stroke and its subtypes: correlation with clinical outcome

Studies in different populations have shown that ischemic stroke can trigger an acute phase response resulting in a rise of plasma concentration of C-reactive protein (CRP). However, there are very limited studies on CRP and first ischemic stroke divided into subtypes. High levels of CRP may also be associated with poor outcome. The present study was taken up to investigate the prognostic value of CRP within 24 h of onset of ischemic stroke. Five hundred and eighty one patients with first stroke and 575 age- and sex-matched healthy controls were involved in the study. High-sensitivity C-reactive protein (hsCRP) levels were estimated, and follow-up interviews were conducted with patients at 3, 6, and 12 months post-event to determine stroke outcome. In addition to this plasma, NO _x (nitrate and nitrite) was measured to detect the serum NO (an important biomarker of inflammation and oxidative stress) levels in ischemic stroke patients and controls. The relationship between CRP value and poor outcome (>2 on modified Rankin Scale Score and <5 on an extended Glasgow outcome scale) was studied. There was a significant association between elevated levels of CRP and NO with the disease. A stepwise multiple logistic regression analysis confirmed these findings after adjustment for potential confounders [adjusted odds ratio = 2.890, 95% CI (1.603–5.011) with p < 0.01 and adjusted odds ratio = 2.364, 95% CI (1.312–3.998) with p < 0.01 for hsCRP and NO, respectively]. After adjustment of potential confounders, patients with high CRP levels had a significant increased risk of poor outcome [adjusted odds ratio = 3.50, 95% CI (1.312–6.365) and p < 0.001]. Elevated levels of hsCRP associated significantly with all stroke subtypes classified according to Trial of ORG 10172 in Acute Stroke Treatment classification except for lacunar stroke and stroke of other determined etiology. In conclusion, hsCRP and NO levels predict the incidence of ischemic stroke and hsCRP is an independent prognostic factor of poor outcome at 3 months.     

Overexpression of carbonic anhydrase IX (CAIX) is an independent unfavorable prognostic marker in endometrioid ovarian cancer

Carbonic anhydrase IX (CAIX) is a strictly membranous expressed metalloenzyme involved in cell adhesion, pH homeostasis, and cancer progression. This study was designed to assess the role of CAIX in primary ovarian cancer. Two hundred five well-characterized primary ovarian carcinomas were analyzed on a tissue microarray. CAIX expression was determined by immunohistochemistry using a four-step scoring system. Moderate and strong membranous CAIX expression was found in 37 out of 205 (18%) of all assessable ovarian cancer specimens. High levels of CAIX expression were related to mucinous and endometrioid phenotype of ovarian carcinomas (p < 0.05). There was no association between CAIX overexpression and tumor stage, grading, and mitotic count of ovarian carcinomas (p > 0.05). In univariate Cox regression analysis, advanced tumor stage (p < 0.01), high tumor grade (p = 0.017), high mitotic count (p = 0.025), and high CAIX expression levels (p = 0.031) were correlated to shorter overall patient survival. High pT stage (p = 0.036) and CAIX overexpression were connected to poor clinical outcome in endometrioid ovarian carcinomas. Multivariate Cox regression hazard analysis comprising tumor stage, tumor grade, mitotic count, and CAIX expression revealed pT2/3 stage and CAIX overexpression (scores 2 and 3) as independent prognostic markers in ovarian cancer (p < 0.01, each) as well as in the subgroup of endometrioid carcinomas (p < 0.05, each). In conclusion, CAIX is overexpressed in a substantial proportion of mucinous and endometrioid ovarian carcinomas and connected to poor patient outcome. Our data support the potential therapeutic benefit of newly developed targeting antibodies in advanced ovarian cancer.     

Comparison of T1 relaxation times in adipose tissue of severely obese patients and healthy lean subjects measured by 1.5 T MRI

Subcutaneous (SAT) and visceral adipose tissue (VAT) differ in composition, endocrine function and localization in the body. VAT is considered to play a role in the pathogenesis of insulin resistance, type 2 diabetes, fatty liver disease, and other obesity‐related disorders. It has been shown that the amount, distribution, and (cellular) composition of adipose tissue (AT) correlate well with metabolic conditions. In this study, T₁ relaxation times of AT were measured in severely obese subjects and compared with those of healthy lean controls. Here, we tested the hypothesis that T₁ relaxation times of AT differ between lean and obese individuals, but also between VAT and SAT as well as superficial (sSAT) and deep SAT (dSAT) in the same individual. Twenty severely obese subjects (BMI 41.4 ± 4.8 kg/m²) and ten healthy lean controls matched for age (BMI 21.5 ± 1.9 kg/m²) underwent MRI at 1.5 T using a single‐shot fast spin‐echo sequence (short‐tau inversion recovery) at six different inversion times (T_I range 100–1000 ms). T₁ relaxation times were computed for all subjects by fitting the T_I‐dependent MR signal intensities of user‐defined regions of interest in both SAT and VAT to a model function. T₁ times in sSAT and dSAT were only measured in obese patients. For both obese patients and controls, the T₁ times of SAT (275 ± 14 and 301 ± 12 ms) were significantly (p < 0.01) shorter than the respective values in VAT (294 ± 20 and 360 ± 35 ms). Obese subjects also showed significant (p < 0.01) T₁ differences between sSAT (268 ± 11 ms) and dSAT (281 ± 19 ms). More important, T₁ differences in both SAT and VAT were highly significant (p < 0.001) between obese patients and healthy subjects. The results of our pilot study suggest that T₁ relaxation times differ between severely obese patients and lean controls, and may potentially provide an additional means for the non‐invasive assessment of AT conditions and dysfunction. Copyright © 2014 John Wiley & Sons, Ltd.     

Mixed-type gastric carcinomas exhibit more aggressive features and indicate the histogenesis of carcinomas

To investigate the pathobiological behaviors of gastric mixed-type (MT) carcinomas and gastric carcinogenesis, the clinicopathological characteristics of MT carcinomas were analyzed and compared with intestinal-type (IT) and diffuse-type (DT) carcinomas. The expression of Ki-67, caspase-3, p53, fragile histine triad (FHIT), maspin, extracellular matrix metalloproteinase inducer (EMMPRIN), vascular growth factor (VEGF), MUC-2, 4, 5AC and 6, CD44, E-cadherin, β-catenin, and phosphorylated glycogen synthase kinase 3β-ser⁹ (P-GSK3β-ser⁹) was examined on tissue microarrays using immunohistochemistry. It was found that MT carcinomas exhibited large size, deep invasion, frequent local invasion, and lymph node metastasis in comparison with IT and DT carcinomas (p < 0.05). All the markers except MUC-5AC showed higher expression in IT than DT carcinomas (p < 0.05). The expression of maspin, EMMPRIN, VEGF, MUC-4, and membrane E-cadherin was stronger in MT intestinal than diffuse component (p < 0.05). Immunoreactivities to Ki-67, EMMPRIN, and VEGF were weaker in IT carcinoma than in the MT intestinal portion (p < 0.05), while the opposite was true for CD44, MUC-2, and MUC-6 (p < 0.05). The MT diffuse component displayed a higher expression of FHIT, VEGF, and P-GSK3β-ser⁹ than DT carcinoma (p < 0.05). The accumulative survival rate of the IT carcinoma patients was higher than the other types (p < 0.05). The invasive depth, venous invasion, lymph node, peritoneal or liver metastasis, and Lauren's classification were independent prognostic factors for gastric carcinomas (p < 0.05). These findings suggested that MT carcinomas were also indicated to be more aggressive than IT and DT carcinomas. Significant differences were observed in the proliferation, apoptosis, angiogenesis, mucin secretion, and cell adhesion between IT and DT carcinomas, whereas only a few of these characteristics showed differences between the MT intestinal and diffuse parts, thus suggesting that both the MT components might originate from the stem cells with similar genetic traits, but follow different histogenic pathways.     

In Vivo Effects of Sequential Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) and Interleukin-2 (IL-2) on Circulating Dendritic Cells (DC) in Patients with Surgically Resected High Risk Cutaneous Melanoma

Dendritic cells were assayed repeatedly in the peripheral blood of consenting melanoma patients receiving adjuvant biotherapy for high risk (stages IIb-IV) melanoma. Postoperatively, adjuvant biotherapy consisted of granulocyte-macrophage colony-stimulating factor [125 μg/m²/day] for 14 consecutive days, followed by interleukin-2 [9 million IU/m²/day] for the next 4 days, and then no treatment for 10–12 days. This was repeated monthly for six cycles. Although white blood cell counts increased, there was no significant elevation in dendritic cell counts during therapy of eleven patients. Within the first cycle during granulocyte-macrophage colony-stimulating factor treatment of seven patients, the absolute DC count decreased (p < 0.04), the percentage of myeloid BDCA-1+ BDCA-2− dendritic cells was significantly lower than baseline (p < 0.003) and the percentage of plasmacytoid BDCA-1− BDCA-2+ dendritic cells was significantly higher than baseline (p < 0.009). Our data suggest mechanisms of potential anti-tumor responses in patients receiving systemic sequential granulocyte-macrophage colony-stimulating factor and interleukin-2 do not include a cumulative gain in peripheral dendritic cell counts or an increase in myeloid BDCA-1+ BDCA-2− dendritic cell subset in the peripheral blood.     

Haematological and clinical chemistry findings associated with sub-acute contamination of drinking water with varied low percentages of used engine oil

This study evaluated the haematology and clinical chemistry profile of rats given drinking water contaminated with varied low percentages of used engine oil (UEO) for a period of 21 days. Fifty female albino rats of 6–7 weeks of age were used for the study. They were divided into five groups (A–E) and given water contaminated with 5%, 1%, 0.1%, 0.01% and 0% vol/vol. of UEO respectively as the only source of drinking water for 21 days. The group E given uncontaminated water (0% contamination) served as the control. The haematological parameters and clinical chemistry profile of the rats was comprehensively evaluated after the 21 days of administration of the group-specific waters. Results showed that contamination of water with up to 5% UEO led to no significant effects (p > 0.05) on all the haematological indices and on the levels of serum alanine amino transferase, aspartate amino transferase, albumin, creatinine and calcium, blood urea nitrogen and fasting blood glucose level, feed consumption and body weight. However, the rat group given water contaminated with 5% UEO had a significantly increased serum alkaline phosphatase (AP) (p < 0.01), total bilirubin (p < 0.05) and cholesterol (p < 0.01), and a significantly decreased serum total protein and globulin (p < 0.01), and water consumption (p < 0.05). The rat group given water contaminated with 1% UEO had a significantly increased serum AP (p < 0.01), total bilirubin (p < 0.05) and cholesterol (p < 0.01), and a significantly decreased water consumption (p < 0.05), while the rat group given water contaminated with 0.1% UEO had a significantly elevated (p < 0.01) serum AP. It was concluded that sub-acute contamination of drinking water of rats with up to 5% UEO led to hepato-biliary disorders and adverse effects on hepatic secretion and excretion, including diminution of serum protein and globulin levels and elevation of serum cholesterol levels, but did not lead to any significant effects on haematology, hepatocellular integrity, kidney/renal function, pancreatic function and body weight.     

High pentraxin-3 plasma levels associate with thrombocytopenia in acute Puumala hantavirus-induced nephropathia epidemica

Our aim was to investigate whether plasma levels of the long pentraxin-3 (PTX3) associate with the severity of Puumala hantavirus-induced nephropathia epidemica (NE). Sixty-one prospectively identified consecutively hospitalized NE patients were examined. Plasma PTX3, interleukin (IL)-6, terminal complement complex SC5b-9, complement component C3, C-reactive protein (CRP), creatinine, sodium, kynurenine, and tryptophan levels, as well as the blood cell count, were determined for up to five consecutive days after hospitalization. Receiver operating characteristic (ROC) analysis revealed that the maximum PTX3 level >101.6 ng/ml (high PTX3) showed a sensitivity of 71% and a specificity of 89% for detecting platelet level <50 × 10⁹/l, with an area under the curve (AUC) value of 0.78 (95% confidence interval [CI] 0.63–0.94). High PTX3 level was also associated with several other variables reflecting the severity of the disease: patients with high PTX3 level had higher maximum blood leukocyte (16.1 vs. 9.7 × 10⁹/l, p < 0.001), plasma IL-6 (16.9 vs. 9.0 pg/ml, p = 0.007), and creatinine (282 vs. 124 μmol/l, p = 0.007) levels than patients with low maximum PTX3 level. They also had longer hospital stays (8 vs. 5 days, p = 0.015) compared to patients with low PTX3 level. High plasma PTX3 levels are associated with thrombocytopenia and the overall severity of NE.