Circulating Heparan Sulfate Proteoglycans as Biomarkers in Health and Disease.

Cell-surface heparan sulfate proteoglycans (HSPGs) play key roles in regulating cell behavior, cell signaling, and cell matrix interactions in both physiological and pathological conditions. Their soluble forms from glycocalyx shedding are not merely waste products, but, rather, bioactive molecules, detectable in serum, which may be useful as diagnostic and prognostic markers. In addition, as in the case of glypican-3 in hepatocellular carcinoma, they may be specifically expressed by pathological tissue, representing promising targets for immunotherapy. The primary goal of this comprehensive review is to critically survey the main findings of the clinical data from the last 20 years and provide readers with an overall picture of the diagnostic and prognostic value of circulating HSPGs. Moreover, issues related to the involvement of HSPGs in various pathologies, including cardiovascular disease, thrombosis, diabetes and obesity, kidney disease, cancer, trauma, sepsis, but also multiple sclerosis, preeclampsia, pathologies requiring surgery, pulmonary disease, and others will be discussed.

Expression and Localization of Glycosaminoglycans/Proteoglycan in Pterygium: An Immunohistochemical Study.

Pterygium is a triangle-shaped fibrovascular hyperplasia of the bulbar conjunctiva on the cornea. The purpose of this study was to analyze Proteoglycans (PGs) by Immunohistochemistry (IHC) in pterygium tissues and to compare the results with normal conjunctiva. Twenty-four patients (14 males) undergoing primary pterygium excision and 17 healthy individuals (10 males), undergoing extracapsular cataract surgery, were included. Pterygium tissues and normal conjunctiva tissues were surgically removed. The tissue sections were fixed in 2% paraformaldehyde and incubated with monoclonal antibodies against PGs anti-mouse IgG. Immunohistochemical study showed stronger expression of keratan sulfate in the stroma of the pterygium compared to normal conjunctiva. An increased expression of heparan sulfate was observed in the epithelial layer and around the pterygium vessels. On the other hand, dermatan sulfate showed an increased expression and localization not only in the sub-epithelial area of the pterygium and normal conjunctiva, yet throughout the stroma of the pterygium. The differences in the expression and localization of the studied extracellular matrix proteoglycans in the pterygium tissue compared to normal conjunctiva may explain the tissue hyperplasia, structure, and the functional properties in pterygium.

Feasibility and effectiveness of inhaled carboplatin in NSCLC patients.

BACKGROUND: Inhaled chemotherapy is under investigation as an alternative therapeutic modality for Non-Small Cell Lung Cancer. METHODS: 60 NSCLC patients were randomized into 3 groups in this study. 20/60 patients (group A-control group) received I.V. chemotherapy (carboplatin AUC ≈ 5.5 D1); 20/60 (group B) received 2/3 of I.V. predicted carboplatin dose by I.V. infusion and the rest 1/3 as aerosol (jet nebulised D1); and 20/60 (group C) received all the predicted I.V. dose of carboplatin as aerosol in 3 equally divided fractions D1-3. In all patients I.V. docetaxel 100/m(2) was as well administered (D1). Lung functional tests were performed in all groups before chemotherapy in the 3rd and 6th cycles. RESULTS: Group B had a statistically significant increase in survival compared to control group A [275 days (95% CI 249-300) vs. 211 (95% CI 185-236)]. In regard to lung functional tests, a statistically significant decline was observed only in FEV1 of group C in 6 months compared to the initial measurement. CONCLUSIONS: Inhaled carboplatin could be given as an alternative root of pulmonary drug delivery in selected patients, but further randomized studies remain to prove whether the inhaled chemotherapy is an efficient and safe treatment modality.

Bronchial asthma, chronic obstructive pulmonary disease and NF-kappaB.

Respiratory diseases place a considerable burden on global health. Bronchial asthma describes many heterogeneous clinical phenotypes that result in chronic bronchial inflammation. Chronic obstructive pulmonary disease (COPD) is one of the most common adult respiratory disorders characterized by chronic airflow limitation that is not fully reversible and is associated with an abnormal inflammatory response of the lungs to noxious particles and gases. Recognition of the global importance and rising prevalence of these diseases and the absence of effective treatments has led to concerted efforts to improve the efficacy of the existing drugs and develop new ones that target cellular and molecular mechanisms that underlie disease pathogenesis. The transcription factor nuclear factor kappa B (NF-kappaB) regulates the expression of a wide array of genes that are involved in the molecular pathobiology of the lung by regulating cellular immune responses, cell adhesion, differentiation, proliferation, angiogenesis and apoptosis. In this work, we review published clinical and experimental studies that link the inhibition of NF-kappaB activity with the treatment of asthma and COPD. Our end point is to help identify pathway-specific inhibitors of NF-kappaB that can be used for the treatment of specific human ailments.

Expression of TNF-alpha, IL-1beta, and IFN-gamma in Staphylococcus epidermidis slime-positive experimental endophthalmitis is closely related to clinical inflammatory scores.

PURPOSE: The purpose of this study was to investigate the expression of inflammatory cytokines TNF-alpha, IL-1beta, and IFN-gamma in the vitreous after experimentally induced endophthalmitis by a Staphylococcus epidermidis slime-producing strain. METHODS: Seventy-two experimental Lewis rats received an intravitreal injection of 7000 viable organisms of Staphylococcus epidermidis slime-producing ATCC strain 35983, while 72 control rats received an intravitreal injection of sterile normal saline. Eyes were graded daily for signs of clinical inflammation and were removed 6, 12, 24, 48, 72 h, and 7 days after injection. Vitreous was obtained and titers of TNF-alpha, IL-1beta, and IFN-gamma were measured with established enzyme-linked immunosorbent assays. RESULTS: In the experimental group, the clinical inflammatory score reached maximum (4+) within 24 h, while inflammation was almost abolished by day 7 (score 0-0.5+). Statistically increased levels of TNF-alpha and IL-1beta were detected in the experimental vitreous with maximum levels observed at 12 h. IFN-gamma was also detected in the experimental vitreous and reached maximum levels at 48 h. None of the cytokines examined was detected in sera at any time point in experimental or control rats. CONCLUSIONS: The results of this study suggest that Staphylococcus epidermidis experimental endophthalmitis induces the expression of cytokines TNF-alpha, IL-1beta, and IFN-gamma in the vitreous. The time course of those cytokine expression levels is closely associated to the clinical presentation of this endophthalmitis model.

Clinical value of bone remodelling markers in patients with bone metastases treated with zoledronic acid.

BACKGROUND: Bisphosphonates have an established role in the treatment of bone metastases from a variety of solid tumours. The objective response to anti-resorptive treatment cannot be evaluated by imaging techniques. A number of bone remodelling markers have been associated with bone metastases status; among them, urine and serum levels of N-terminal telopeptide of collagen type I (NTx) seem to have the best diagnostic accuracy. However, serum NTx has not yet been properly evaluated. PATIENTS AND METHODS: Seventy-one consecutive patients with newly diagnosed skeletal metastases were enrolled in this prospective study. All of them were treated with zoledronic acid at 4 mg, every 3 or 4 weeks. Serum NTx and bone-isoform of alkaline phosphatase (BAP) were measured by enzyme-linked immunosorbent assays at baseline and every 2 months thereafter. RESULTS: At baseline, serum NTx and BAP levels were significantly higher in patients with blastic than lytic bone lesions and in those with multiple rather than few bone site involvement. Forty-seven patients were followed for a median period of 139 days. Zoledronic acid resulted in a significant NTx reduction at first and second post-treatment evaluations (mean reduction of 43% at first evaluation); thereafter, mean NTx levels remained suppressed. In contrast, BAP levels did not show any significant changes. Bone disease progression resulted in a significant NTx elevation by an average of 69%. The initial response of NTx to zoledronic acid was correlated with the long-term clinical outcome of bone disease: patients with an initial NTx elevation had a significantly higher rate of bone disease progression compared to those with an initial NTx decline (66.7% versus 18.8%, p=0.001). Extraskeletal disease or bone irradiation did not influence NTx response. CONCLUSION: Serum NTx appears to be a useful marker in monitoring patients with skeletal metastases, as it is correlated with the type and bulk of bone disease and reflects bone disease progression. It is also useful in monitoring bisphosphonate therapy, while the initial response to this therapy seems to bear a prognostic significance for bone disease outcome.

Diagnostic value of bone remodeling markers in the diagnosis of bone metastases in patients with breast cancer.

Metastatic spread to bone is common in patients with breast cancer and its early detection is required for the better management of these patients. Several biochemical markers of bone remodeling have been recently developed, in order to assess metastatic bone disease with non radiologic methods. The pyridinolin cross-linked amino-terminal telopeptide of type I collagen (NTx) has been measured in serum and urine as a specific marker of bone collagen breakdown, while the bone-isoform of alkaline phosphatase (BAP) has been used to determine bone formation activity. Thirty-three consecutive ambulatory patients with metastatic breast cancer and bone metastases and 31 with extraskeletal metastases only, matched for age and menopausal status, were studied. Serum levels of NTx and BAP were measured by enzyme-linked immunosorbent assays. The diagnostic accuracy of both markers was evaluated by receiver operating characteristic (ROC) analysis. Patients with bone metastases had significantly higher levels of NTx (37.0+/-36.9 nM BCE versus 23.5+/-21.0 nM BCE, P<0.05) and BAP (57.8+/-31.7 U/L versus 36.5+/-28.5 U/L, P<0.01) compared to those without bone metastases. NTx was positively correlated with BAP (R=0.340, P<0.01). The area under the ROC curve was 0.671 for NTx and 0.755 for BAP. Using a cut-off value of 29.7 nM BCE for NTx, specificity and sensitivity were 87.1% and 45.5%, respectively; in the case of BAP, using a cut-off value of 50.6 U/L, the specificity and sensitivity were 90.3% and 54.5%, respectively. In patients not receiving concomitant hormonal treatment, the area under the ROC curve was 0.724 for NTx and 0.822 for BAP; in this subgroup of patients, using a cut-off value of 30.0 nM BCE for NTx, the specificity and sensitivity were 96.2% and 47.1%, respectively, while using a cut-off value of 50.0 U/L for BAP, the corresponding percentages were 92.3% and 70.6%. Although serum NTx and BAP are quite specific, they are not sensitive enough to diagnose bone metastases in patients with advanced breast cancer. Their diagnostic accuracy, however, is considerably enhanced in patients not receiving hormonal therapy.