Biological therapy downregulates the heterodimer S100A8/A9 (calprotectin) expression in psoriatic patients.

The pathophysiology of psoriasis is very complex and involves an interplay between immune cells and keratinocytes. The keratinocyte production of calprotectin (S100A8/A9), induced by the inflammatory psoriatic milieu, may be involved in initiating immune cell invasion, as well as in propagating inflammation. However, the exact role of calprotectin in psoriasis remains unclear. Therapeutic approaches utilizing adalimumab, etanercept and ustekinumab are widely used in psoriatic treatment, but their anti-inflammatory mechanisms are not fully understood. The aim of this study was to investigate, by immunohistochemical analysis, the expression of the heterocomplex S100A8/A9 in lesional skin from psoriatic patients undergoing biological therapy with adalimumab, etanercept or ustekinumab. Our results showed that S100A8/A9, absent or present at very low level in skin biopsies from healthy subjects, is dramatically upregulated in each epidermal layer from psoriatic patients. Interestingly, calprotectin was mainly localized in keratinocyte nuclei from psoriatic patients, suggesting a role of S100A8/A9 in keratinocyte nuclear function. Furthermore, we have shown that the biological treatment induced a drastic reduction of S100A8/A9 expression in skin biopsies from treated patients, correlating with PASI reduction. Our results suggest that calprotectin may play a crucial role as a significant marker of inflammation in psoriasis, and that its reduction of expression may be considered a favourable prognostic marker in psoriasis.

Effect of levofloxacin treatment on semen hyperviscosity in chronic bacterial prostatitis patients.

Changes in seminal fluid viscosity (SFV), reactive oxygen species (ROS) production, cytokines and seminal leucocyte concentration related to microbiological outcome in patients with chronic bacterial prostatitis (CBP) were studied. One hundred and ten infertile patients with CBP (positive sperm culture ≥10(5) colony-forming units [CFU] ml(-1), pathogens or Chlamydia in expressed prostatic secretions) were treated with levofloxacin 500 mg daily for 14 consecutive days per month for 3 months. In case of bacterial prostatitis, two conditions were examined: responders, eradication of 0 to <10(3) CFU ml(-1) (n = 78) and poor responders, >10(3) to <10(5) CFU ml(-1) (n = 32). Compared with poor responders, responders showed a significant increase of sperm progressive motility and a significant decrease in seminal leucocyte count, SFV, liquefaction time, ROS production (in all fractions and conditions), seminal tumour necrosis factor-α and interleukin 6. None of these variables showed significant differences compared with a control group of 37 fertile men. On the other hand, the poor responders showed significant changes in these variables compared with matched pretreatment values. In patients with CBP, antibiotic therapy alone leads to eradication in ≈71%, with improvement of sperm progressive motility, SFV and the framework of prooxidative factors. However, in the remaining ≈29% with poor antibiotic responsiveness, a deterioration of all variables is observed.

Matrix metalloproteinases and their inhibitors as markers of inflammation and fibrosis in chronic liver disease (Review).

Chronic liver disease (CLD) is a cause of morbidity and mortality worldwide, due to haemodynamic and metabolic complications of liver cirrhosis. During CLD the extracellular matrix undergoes a process of remodelling, leading to new collagen formation and deposition. Tissue remodelling is regulated by fine molecular mechanisms, involving proteases, inhibitors and growth factors. The major role in matrix degradation is played by matrix metalloproteinases (MMPs), a class of zinc and calcium-dependent enzymes, and their tissue inhibitors (TIMPs). Along with the progress in diagnostic techniques, leading to more precise and less invasive methods, the concept of monitoring has gained importance for the clinical management of CLD. At the present state of our knowledge, liver biopsy still represents an essential procedure for staging liver disease. However, despite its importance, liver biopsy presents some limitations: the risk of a disease underestimation is the most significant one, as hepatic lesions are often irregularly located within the liver. Parallel to the limitations of liver biopsy, clinical needs for an early identification of progressive fibrosis require additional non-invasive techniques to be developed. In this review we discuss the major problems concerning this important clinical necessity. Moreover, we focus on the role of MMPs and TIMPs in the pathogenesis of CLD, as well as their possible use as non-invasive serum markers for inflammation and fibrosis in this pathology.

Meaning of elevated CA 19-9 serum levels in chronic hepatitis and HCV-related cirrhosis.

AIM: Carbohydrate 19-9 antigen (CA 19-9) has been used in the diagnosis and follow-up of gastrointestinal tumors. However, a remarkable reduction of specificity has been described in subjects with chronic diseases. Elevated CA 19-9 serum levels have been described in non neoplastic liver diseases, such as hepatic cirrhosis, where they correlate with the fibrosis grade and the disease severity. The aim of the study is to evaluate CA 19-9 levels in chronic hepatitis patients (CH) and hepatic cirrhosis patients, Hepatitis C Virus (HCV)-correlated. Our goal was to establish whether elevated CA 19-9 levels can be considered a non casual event in chronic liver disease and whether a correlation can be found between CA 19-9 levels and the severity of the disease. METHODS: 116 patients have been recruited (76 m, 40 f, average 54 years); 56 patients were affected by CH and 60 by hepatic cirrhosis (Child A). All patients were HCV+, genotype 1b. Patients positive to CA 19-9 high levels were subjected to abdominal echography, EGDS, colonscopy, abdominal CT. RESULTS: Fifty two percent presented high levels of CA 19-9. None was affected by intestinal or pancreatic neoplasia, or colestatic icterus. CA 19-9 levels were elevated in 46% of patients with chronic hepatitis, and in 54% in patients with hepatic cirrhosis. Furthermore, CA 19-9 levels in hepatic cirrhosis compared to CA 19-9 levels in chronic hepatitis was statistically significant (P>0.007). CONCLUSION: Increased serum levels of CA 19-9 are frequent in chronic viral hepatitis; this often does not indicate a contemporary neoplastic disease and correlates in a statistically significant way (P>0.007) with the severity of the disease.

Analysis of G(-174)C IL-6 polymorphism and plasma concentrations of inflammatory markers in patients with type 2 diabetes and peripheral arterial disease.

AIMS: To determine whether the G(-174)C interleukin 6 (IL-6) polymorphism influences the development of peripheral arterial disease (PAD) in individuals with type 2 diabetes. This was investigated by comparing the distribution of G(-174)C genotypes between patients with type 2 diabetes and PAD (PAD+) and those with type 2 diabetes but without PAD (PAD-). Plasma concentrations of IL-6, fibrinogen, C reactive protein (CRP), and vascular endothelial growth factor (VEGF) were also compared in PAD+ and PAD- patients. METHODS: Blood samples were collected from 146 PAD+ and 144 PAD- patients. SfaNI was used to determine the G(-174)C genotype. Plasma concentrations of IL-6, fibrinogen, CRP, and VEGF were measured by an enzyme linked immunosorbent assay. RESULTS: The GG genotype was more common in PAD+ patients than in PAD- patients. PAD+ patients also had increased mean plasma concentrations of IL-6, fibrinogen, CRP, and VEGF compared with PAD- patients. Mean plasma concentrations of IL-6, fibrinogen, and CRP in both PAD+ and PAD- patients were higher in those with the GG genotype than in those with the GC or CC genotypes. In contrast, mean plasma concentrations of VEGF in PAD+ and PAD- patients were not significantly different between those with different G(-174)C genotypes. CONCLUSIONS: These results support a model in which the GG genotype promotes PAD development among individuals with type 2 diabetes by inducing increased release of IL-6. Higher concentrations of IL-6 among those with the GG genotype is associated with increased plasma concentrations of fibrinogen and CRP.

Analysis of hepatitis C virus infection among health-care workers: an observational study.

AIM: Hepatitis C virus (HCV) is one of the most common blood-borne pathogens transmitted from patients to health care workers (HCWs). The Centers for Disease Control and Prevention (CDC) have developed a set of universal precautions to help prevent transmission of blood-borne pathogens between patients and HCWs in health care settings. HCV infection status among HCWs and proportion of HCWs experiencing occupational blood exposure accidents were monitored to assess the risk of HCV infection among HCWs at a hospital in Catania, Italy. METHODS: The number of HCWs reporting occupational blood exposure accidents during 1999 and 2004 were compared to examine whether there was any change in the incidence of these accidents among 900 HCWs. HCV infection status of these HCWs was also analyzed in 1999 and 2004 to determine how many were infected with HCV during this time period. RESULTS: HCV infection was detected in 21 out of 900 subjects in 1999. The remaining 879 HCWs remained HCV-negative until they were last tested in 2004. There was a statistically significant decrease in the number of HCWs that experienced occupational blood exposure accidents from 306 in 1999 to 240 in 2004 (P = 0.001). CONCLUSIONS: The finding that all 871 HCV-negative HCWs remained HCV-negative from 1999 until 2004 supports the view that the set of universal precautions recommended by the CDC are helpful for preventing HCV transmission from patients to HCWs. HCWs must continue following these precautions to prevent transmission of HCV and other blood-borne pathogens between patients and HCWs in the future.

Frequency of bcl-2/IgH translocation in patients with non-Hodgkin's lymphoma and chronic hepatitis C virus infection.

AIM: It has been previously suggested that t(14;18) translocation of bcl-2 to the immuno-globulin heavy chain (IgH) locus may contribute to pathogenesis of lymphoproliferative disorders related to hepatitis C virus (HCV) infection, including type II mixed cryoglobulinemia (MC). METHODS: In this study, the presence or absence of t(14;18) translocation was determined in tumor biopsy specimens and peripheral blood mononuclear cells (PBMCs) for 48 NHL patients with chronic HCV infection. RESULTS: In tumor biopsy specimens from 32 HCV-positive NHL patients, bcl-2/IgH translocation was detected in 1 of 13 patients with MC syndrome (7.7%) and 3 of 19 patients without MC syndrome (15.8%). In PBMCs from 23 HCV-positive NHL patients, this translocation was observed in 3 of 6 patients with MC syndrome (50%) and 4 of 17 patients without MC syndrome (23.5%). Interestingly, bcl-2/IgH translocation was found in 2 extranodal marginal zone B-cell lymphoma tissues from HCV-infected patients. CONCLUSIONS: However, additional studies are required to better clarify the relationship between this translocation and extranodal marginal zone B-cell lymphoma development. Although the frequency of bcl-2/IgH translocation in PBMCs from patients with chronic HCV infection is higher than that of other NHL patients, this increased translocation rate remains to be elucidated.

Persistent high plasma levels of interleukins 18 and 4 in children with recurrent infections of the upper respiratory tract.

The aim of this study was to examine whether children with recurrent infections of the upper respiratory tract might have alterations in the systemic immune response to viral infections as compared with healthy control children. We quantitated plasma levels of interferon-gamma, interleukin-12, interleukin-18, interleukin-4, lymphocyte subpopulations, serum immunoglobulins, and subclasses of immunoglobulin G in 30 children under the age of 6 years with recurrent infections of the upper respiratory tract, both during the acute phase of the infection and 4 weeks later, when clinical symptoms had resolved, as well as in 20 normal controls. We found elevated levels of immunoglobulin G primarily due to increased levels of immunoglobulin G(1). Moreover, significantly higher levels of interleukin-18 and interleukin-4 were noted during the acute phase of infection among children with an increased incidence of respiratory infections as compared with the controls (P =.022 and P =.0001, respectively), while plasma levels of interferon-gamma and interleukin-12 were significantly lower (P =.034 and P =.0001, respectively) than in controls. We suggest that an imbalance between T-cell helper type-1 and T-cell helper type-2 immune responses might be responsible for the perpetuation of recurrent infections of the upper respiratory tract.

Cell death in human acute myelogenous leukemic cells induced by pyrrolidinedithiocarbamate.

Pyrrolidinedithiocarbamate (PDTC) is a metal chelating compound, which exerts both pro-apoptotic effect and pro-oxidant activity on many cells. Our objective was to investigate whether PDTC was able to interfere with apoptotic process in leukemic and normal bone marrow CD34+ cells. Since hematopoietic growth factors stimulate growth and differentiation and prevent apoptosis, we therefore studied the effect of growth factors pretreatment, such as interleukin-3 and granulocyte-macrophage colony stimulating factor, in human myeloid CD34+ cells to evaluate whether they protect the cells from the apoptotic action of PDTC. We revealed that PDTC exerted an apoptotic effect in leukemic CD34+ cells. This effect was dependent on the ability of this compound to generate the oxidation of cellular glutathion to its disulphide and consequently to induce an intracellular oxidative stress. Hematopoietic growth factors did not protect cells from apoptosis induced by previous treatment with PDTC. The ability of PDTC to induce apoptosis was restricted to acute myelogenous leukaemia CD34+ cells, since normal CD34+cells were insensitive to the pro-oxidant effect of PDTC. These findings imply that normal cells are equipped with mechanisms by which they respond differently to PDTC effects with respect to leukemic cells.

Autosomal recessive hypercholesterolemia in a Sicilian kindred harboring the 432insA mutation of the ARH gene.

We describe a Sicilian family presenting a recessive form of hypercholesterolemia harboring a mutation of the autosomal recessive hypercholesterolemia (ARH) gene. In two of the three sibs, a 26-year-old male and a 22-year-old female, a severe hypercholesterolemia was diagnosed with very high levels of plasma cholesterol (15.9 and 12.2 mmol/l, respectively); tendon xanthomatas and xanthelasms were present and in the male proband was documented a diffuse coronary atherosclerotic disease with a rapid and fatal progression. Both the parents had normal or slightly increased levels of plasma cholesterol. All causes of secondary hypercholesterolemia were ruled out as well as an involvement of the LDL receptor or apoB genes. Beta-Sitosterol plasma levels were in the normal range. Cultured fibroblasts from skin biopsy from parents and the two probands displayed a normal ability to bind and degrade 125I-LDL. Direct sequencing of ARH gene demonstrated the presence of a 432insA mutation in homozygosis in the two probands; parents were heterozygotes for the same mutation. This mutation is the first report of a mutation of the ARH gene responsible for recessive forms of hypercholesterolemia in Sicily.

Serum concentrations of the interferon-gamma-inducible chemokine IP-10/CXCL10 are augmented in both newly diagnosed Type I diabetes mellitus patients and subjects at risk of developing the disease.

AIMS/HYPOTHESIS: Chemokines are chemotactic cytokines controlling the recruitment of leukocytes from the blood by regulating integrin adhesiveness. It has been shown that the migration of CD4+Th1 and CD4+Th2 cells is governed by specific chemokines. Increasing evidence suggests that the CD4+Th1 cheomoattractant chemokine CXCL10, also termed Interferon (IFN)-gamma -inducible protein (IP)-10 is pathogenetically involved in several immunoinflammatory and autoimmune diseases. METHODS: IFN-gamma and IP-10 were quantified by solid-phase ELISA in sera of patients with either newly diagnosed or long-term Type I (insulin-dependent) diabetes mellitus, and in sera of their healthy first degree relatives. The latter were subdivided into "low" and "high" risk prediabetic subjects depending on whether they were negative or positive for the anti-beta-cell autoantibodies ICA and GAD. RESULTS: Compared with healthy control subjects (18%, 9/50), those with a low risk of disease (21%, 5/24) and the group of patients with long-term Type I diabetes (24%, 12/50), IP-10 was found more frequently and at increased concentrations in both newly diagnosed Type I diabetic patients (84%, 42/50) and in those with a high risk of disease (73%, 16/22); in the latter, the IP-10 concentrations correlated with those of IFN-gamma. CONCLUSION/INTERPRETATION: Circulating IP-10 concentrations is increased in patients with Type I diabetes, but only during the early and subclinical stage of the disease.

Effects of ischaemic stress on leukocyte activation processes in patients with chronic peripheral occlusive arterial disease: role of L-propionyl carnitine administration.

An open study was carried out to assess whether, in patients with occlusive peripheral arterial disease (PAD), ischaemic stress induced by maximal physical exercise is associated with leukocyte activation processes, and to evaluate the effects of L-propionyl carnitine (LPC) administration on such processes. Fifteen patients with occlusive PAD (stage II-A), with a mean pain-free walking distance (PWD) of 199 +/- 70.66 m were orally treated with 2000 mg/day LPC for 2 months. Serum levels of E-selectin, P-selectin, L-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-I (VCAM-1) were measured at rest and after the performance of a treadmill walking test (treadmill speed 3.5 km h(-1), inclination 12%) in the untreated condition, and again after treatment with LPC. Significant increases of these factors were observed after maximal exercise compared with resting values. Such increase was significantly reduced after LPC treatment compared with the untreated condition. This study shows that ischaemia induced by maximal stress is associated with leukocyte activation processes, and that LPC is capable of modulating these processes. LPC, therefore, may have a protecting role during ischaemia.

Venous stasis causes release of interleukin 1beta (IL-1beta), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNFalpha) by monocyte-macrophage.

BACKGROUND: many studies have investigated between venous stasis, functions of the vascular and perivascular anastomotic structure, venous endothelium and circulating leukocytes. SETTING: patients with varicose veins (n = 15) and a healthy control group (n = 15). METHODS: the authors investigated some soluble mediators of monocytes-macrophages, which induce inflammation. They determined interleukin 1beta (IL-1beta) and interleukin 6 (IL-6) and tumor necrosis factor alpha (TNFalpha) levels at rest and after induced venous occlusion (using an inflated cuff to 60 mmHg for 25 minutes). RESULTS: their results revealed elevated baseline production in the former and that induced venous occlusion further augmented the levels of all cytokines in the study series, especially in patients with varicose veins. CONCLUSION: The authors believe that the study shows functional activation of monocyte-macrophages related to venous stasis as a consequence of venous hypertension. Cell response damages the endothelial structure and may represent an important element in the pathophysiology of chronic venous insufficiency.

[Endothelin-1 variation in the coronary circulation during angioplasty with a stent implant]. / Variazione di endotelina-1 nel circolo coronarico durante angioplastica con impianto di stent.

Endothelin-1 (ET-1) is an endothelium-derived mediator with vasoconstrictive and mitogenic activity which stimulates vascular smooth muscle cell proliferation. The aim of this study was to evaluate ET-1 production during percutaneous transluminal coronary angioplasty (PTCA) and elective stent implantation. We hypothesized that the additional vessel wall trauma induced by stent deployment might be associated with a greater production of ET-1. To this end, ET-1 levels were measured in 18 patients undergoing PTCA and stenting (12 with left anterior descending coronary artery stenosis and 6 with circumflex artery lesion). The sampling sites were the coronary ostium and coronary sinus in basal conditions (before the procedure), during first balloon inflation, and 5, 20, 60 min after the end of first balloon inflation. At baseline, ET-1 levels were higher in the coronary sinus than in coronary ostium (1.58 +/- 0.22 vs 1.29 +/- 0.20 pg/ml, p < 0.001). During first balloon inflation, ET-1 coronary sinus levels significantly diminished with respect to the basal levels (1.08 +/- 0.32 vs 1.58 +/- 0.22 pg/ml, p < 0.001). Further significant variations of ET-1 levels were not detected neither following the first balloon inflation nor after stent deployment. In conclusion, the culprit lesion seems to produce most of ET-1 circulating in the coronary tree. This is demonstrated by higher ET-1 levels in the coronary sinus compared to coronary ostium at baseline, and even more by the significant ET-1 reduction in the coronary sinus during first balloon inflation. Despite our expectations, we did not detect any significant ET-1 increase during stent deployment.

Efficacy of an analysis of lymphocyte subsets in predicting the clinical response to alpha-interferon therapy in thalassaemia patients with chronic infection by hepatitis C virus: a pilot study.

alpha-interferon (alpha-IFN) has been used to treat chronic non-A non-B hepatitis in thalassaemic patients with response rates from 45% to 83%. Unfortunately, treatment with alpha-IFN is associated with side-effects which have a negative effect on the quality of life of the patient. Therefore it would be useful if we could distinguish in advance those patients who would benefit from such therapy from those who would not. In the present study we found that the modification of lymphocyte subsets 20 h after the administration of the first dose of alpha-IFN revealed that relative numbers of T helper lymphocytes (CD4+) increased in three non-responding patients and decreased in five responding patients, whereas those of T suppressor lymphocytes (CD8+), and natural killer cells (CD57+, CD16+) decreased in non-responding patients and increased in responding patients. Therefore analysis of the lymphocyte subsets CD4, CD8, CD57 and CD16 before and 20 h after the administration of alpha-IFN can be used to predict the clinical response to treatment with alpha-IFN.

[Immunologic study on patients with head and neck cancer treated with thymopentin associated with surgery, chemotherapy and radiotherapy]. / Studio immunologico di pazienti con neoplasie ORL trattati con timopentina con terapia adiuvante la chirurgia, la chemioterapia e la radioterapia.

Patients with H&N tumours treated with surgery, chemo- and radiotherapy also underwent an immunologic therapy with timopentina to evaluate clinic and immunologic efficacy during a 1-year follow-up. Twenty-five patients were recorded in this study divided at random into two groups. In group A the patients were administered timopentina (50 mg/3 times per week/6 weeks) subcutaneously in 4 o 5 cycles during the year. Group B were not administered timopentina. The immunologic state was assessed through investigation of the following: Evaluation of PBL and their T and B cell subpopulations Phagocytosis and blastigenesis Surface receptor and soluble receptor of IL2 NK activity IL1 production. The immunologic values of the two groups were correlated against a control group of twenty non-neoplastic patients. Our study revealed a better immunologic conditions at the end of follow-up in patients treated with timopentina compared to the other patients.

Plasma beta-endorphin levels and natural-killer cells in two cases of congenital indifference to pain.

We have investigated plasma beta-endorphin (beta-E), ACTH, and cortisol in two cases of congenital indifference to pain (CIP), a rare syndrome characterized by unresponsiveness to painful stimuli. As the two patients had frequent skin infections, we also studied lymphocyte response to mitogens in the absence or presence of beta-E. In addition, we explored a series of lymphocyte membrane antigens related to different aspects of the immune response, such as CD3+, CD4+, CD8+, B, NK Leu 7, Leu 9, and Leu 19, anti-interleukin-2 receptor (anti-TAC). Plasma beta-E levels in the two patients were significantly higher than in controls, whereas plasma ACTH and cortisol levels were normal. Lymphocyte response to the mitogen phytohemagglutinin was normal. The expression of Leu 7, Leu 9, and Leu 19, three antigens related to natural killer cells, was decreased by about 50%. The results indicate that in the two cases of CIP studied, high plasma beta-E levels are associated with a reduction in the expression of natural killer cells. This suggests that the two phenomena are specifically related to each other. These data represent further evidence of the possible pathophysiological relevance of the neuroendocrine-immune feedback.

Circulating immunocomplexes and laryngeal carcinoma.

Serum levels of circulating immunocomplexes (CIC) were studied in a group of 37 patients with laryngeal carcinoma using two polyethylene glycol-precipitation methods. The preoperative values of this group showed higher levels of CIC when compared with 140 normal controls. No correlation was noted between tumoral stage, oncotype, and serum levels of CIC. Previous studies have shown that the status of several different types of human tumors can be monitored by serial determinations of levels of CIC. We believe that this technique can be used to evaluate the efficacy of therapy and to detect the recurrence of laryngeal carcinoma.

Effects of ketoprofen on the calciuric and uricosuric activities of calcitonin in man.

The action of ketoprofen on the calciuric and uricosuric effects of porcine calcitonin in man is reported. The drug is capable of inhibiting both effects. This action is considered like an effect on CT receptors in the kidney or a competitive action on prostaglandin synthesis.