Medullary thyroid cancer (MTC) is an aggressive malignancy responsible for up to 14% of all thyroid cancer‐related deaths. It is characterized by point mutations in the rearranged during transfection ...(RET) proto‐oncogene. The activated RET kinase is known to signal via extracellular signal regulated kinase (ERK) and phosphoinositide 3‐kinase (PI3K), leading to enhanced proliferation and resistance to apoptosis. In the present work, we have investigated the effect of two serine/threonine‐protein kinase B‐Raf (BRAF) inhibitors (RAF265 and SB590885), and a PI3K inhibitor (ZSTK474), on RET‐mediated signalling and proliferation in a MTC cell line (TT cells) harbouring the RETC634W activating mutation. The effects of the inhibitors on VEGFR2, PI3K/Akt and mitogen‐activated protein kinases signalling pathways, cell cycle, apoptosis and calcitonin production were also investigated. Only the RAF265+ ZSTK474 combination synergistically reduced the viability of treated cells. We observed a strong decrease in phosphorylated VEGFR2 for RAF265+ ZSTK474 and a signal reduction in activated Akt for ZSTK474. The activated ERK signal also decreased after RAF265 and RAF265+ ZSTK474 treatments. Alone and in combination with ZSTK474, RAF265 induced a sustained increase in necrosis. Only RAF265, alone and combined with ZSTK474, prompted a significant drop in calcitonin production. Combination therapy using RAF265 and ZSTK47 proved effective in MTC, demonstrating a cytotoxic effect. As the two inhibitors have been successfully tested individually in clinical trials on other human cancers, our preclinical data support the feasibility of their combined use in aggressive MTC.
Currently, ELISA for detection of anticardiolipin (aCL) and anti-β2glycoprotein I (anti-β2GPI) antibodies is not standardized. Recently, few studies have compared the performance of ELISA with that ...of fluorescence enzyme immunoassay (FEIA), but they have produced debatable results. The aim of this investigation was to compare ELISA with FEIA results in detecting aCL and anti-β2GPI antibodies.
The study cohort included 94 primary antiphospholipid syndrome (PAPS) patients, 65 subjects with the clinical criteria for PAPS classification but ELISA negative for the laboratory criteria and 165 control subjects. Serum IgG/IgM aCL/anti-β2GPI antibodies were determined using FEIA-EliA™ and a home-made ELISA.
The sensitivities of the two methods were similar with the exception of IgM aCL which was found to be significantly higher in the PAPS patients using the ELISA method, even if IgM aCL was detected at a low level by both techniques. The two assays had a comparable specificity, a high/significant agreement and a significant correlation between the antibody levels. FEIA testing uncovered no significant prevalence of any antiphospholipid (aPL) antibody in the ELISA negative patients.
Our results suggest that FEIA is comparable to a home-made ELISA.
•ELISA for detection of aCL and anti-β2GPI antibodies is not standardized yet.•We compare ELISA with FEIA method in detecting aCL and anti-β2GPI antibodies.•A large cohort of patients with primary antiphospholipid syndrome is included.•FEIA antibody results are comparable to those of a home-made ELISA.•Data support the routine use of FEIA in detecting aCL and anti-β2GPI antibodies.
The aim of this study was to evaluate the measurement of C-peptide, insulin, sex hormone binding globulin (SHBG), and dehydroepiandrosterone sulfate (DHEAS), tests of specific clinical value that are ...not frequently measured in routine clinical laboratories, carried out using an automated system, AIA 2000 (Tosoh). The obtained data demonstrated that the evaluated system, characterized by fluorescence system detector provides satisfactory analytical performance comparable to those of different instrument that use the well known and widely diffused chemiluminescent principle (Immulite 2000). These characteristics allow an excellent comparability between methods for the measurement of same specific hormones not frequently used in clinical practice.
Aim of the Study. To investigate the relationships between the adipocytokine levels, markers of inflammation, and vascular remodelling in pregnancies complicated by intrauterine growth restriction ...(IUGR). Materials and Methods. This was a retrospective study. One hundred and forty pregnant patients were enrolled. Adiponectin, leptin, tumor necrosis factor α (TNFα), interleukin-6 (IL-6), and C reactive protein (CRP) were assessed in IUGR, small for gestational age (SGA), and appropriate for gestational age (AGA) mother-child couples at delivery. IUGR and SGA fetuses were defined as fetuses whose estimated fetal weight (EFW) was below 10th percentile for gestational age with and without umbilical artery (UA) Doppler abnormalities, respectively. Fetal aorta intima media thickness (aIMT) was evaluated by ultrasound in the same fetal groups. Data were analyzed by R (version 2.15.2). Results. There were 37 IUGR mother-child couples, 33 SGA, and 70 AGA. Leptin, TNFα, IL-6, and CRP serum levels were higher in IUGR pregnant patients ( P < 0.05 ). Adiponectin levels were significantly reduced in IUGR fetuses compared to SGA and AGA, while leptin, TNFα, and IL-6 levels were higher in IUGR group ( P ≤ 0.05 ). Fetal aIMT was significantly higher in IUGR ( P < 0.05 ) and in this group there was a negative correlation between aIMT and adiponectin/leptin ratio (A/L ratio) ( P < 0.05 ) and between adiponectin and IL-6 levels ( P < 0.05 ). Conclusions. In conclusion, compared to SGA and AGA, IUGR fetuses had reduced circulating levels of adiponectin and elevated measures of aIMT and several inflammatory markers. Moreover, adiponectin levels were negatively correlated with aIMT in IUGR fetuses suggesting a possible causal link between reduced adiponectin and vessel remodelling.
Context: The recently discovered hormone resistin is linked to the development of insulin resistance, but direct evidence of resistin levels in humans with nonalcoholic fatty liver disease (NAFLD) is ...lacking.
Methods: We conducted this study to assess the relationship between serum resistin and NAFLD. We measured serum resistin and biochemical, hormonal, and histological correlates in 28 NAFLD patients, 33 controls, and 30 obese patients body mass index (BMI), >30 kg/m2 without NAFLD.
Results: Resistin and adiponectin expression were measured in sc adipose tissue by quantitative RT-PCR. Resistin was higher in NAFLD patients compared with controls (5.87 ± 0.49 vs. 4.30 ± 0.20 ng/ml; P = 0.002) and obese patients (4.37 ± 0.27 ng/ml; P = 0.002). Increased resistin mRNA was also found in the adipose tissue of NAFLD patients compared with controls and obese subjects.
Conclusions: Both NAFLD and obese patients had lower adiponectin levels, whereas leptin was increased only in the obese group. No correlation was found between resistin and high-sensitivity C-reactive protein, BMI, homeostasis model assessment, insulin, glucose, transaminases, and lipid values. A positive correlation was found between resistin and histological inflammatory score. These data report increased resistin in NAFLD patients that is related to the histological severity of the disease, but do not support a link between resistin and insulin resistance or BMI in these patients.
Abstract
PCOS patients were frequently characterized by lower plasma vitamin D levels. The mechanisms involved in this dysfunction remains still debated, therefore we evaluated the role of androgen, ...insulin and body weight on the serum VitD levels in women with or without PCOS. Eighty one patients 18-42 yrs old were studied into "SUMMER" and "WINTER" seasonal period: thirty seven PCOS, seventeen no-ovarian hyperandrogenic (noPCOS), twelve functional hypothalamic amenorrhea (FHA) and finally fifteen healthy (Con). Patients were further divided into: lean (L), obese (O), normo- (nINS) and hyperinsulinemic (hINS). All hormonal and metabolic parameters were measured at 1-7 days of the menstrual cycle. Our results show that VitD levels were lower in PCOS and in noPCOS than in FHA and Con, in particular in (O) and (hINS) PCOSs. Both in summer and in winter, PCOSs had basal VitD levels significantly lower than FHA and Con, whereas they were similar to noPCOS. Yet, LhINS and OPCOS had VitD levels lower than Con and noPCOS. VitD levels were comparable in LnINS PCOS and Con. In conclusion, PCOSs had levels of VitD lower than controls. Weight and hyperinsulinemia had a significant influence on these values. Finally, over 70% of our healthy patients had VitD deficiency.
In vitro model for IgE mediated food allergy Pizzuti, Daniela; Senzolo, Marco; Buda, Andrea ...
Scandinavian journal of gastroenterology,
02/2011, Letnik:
46, Številka:
2
Journal Article
Recenzirano
Abstract
Background.
In intestinal food allergy, the non-specificity of gastrointestinal symptoms and the limited access to the reacting organ are the reasons for the limited understanding of the ...pathophysiology of this disease and the difficulties in establishing an appropriate diagnosis in the individual patient.
Objective.
To develop an in vitro model reproducing pathophysiological mechanisms of IgE mediated food allergy.
Methods.
Distal duodenum biopsies of nine patients with food allergy and 10 control subjects were cultured for 3 h with medium alone and with 1mg/ml of peptic-tryptic digest of wheat gliadin, wheat albumins, and apple proteins. Each biopsy was used for conventional histological examination and for immunohistochemical detection of IgE-positive cells. We have also analyzed the expression of tight junction proteins, occludin, claudin-1, and ZO-1 by immunoconfocal microscopy. Histamine and tryptase release were measured in the culture medium and collected at 0, 30 min, and 3 h of culture using an enzyme and radio immunoassay, respectively.
Results.
Exposure of small intestinal biopsy specimens of patients with food allergy to food allergens led to a significative increase of IgE-positive cells with a significative increase of histamine and tryptase release and an altered expression of tight junction proteins. No differences were found in intestinal biopsies of controls, cultured with or without food antigens.
Conclusions.
Small intestinal organ culture is a functional model of food allergy and could be considered as an in vitro oral food challenge, with evident reduction of costs and risks for the patients.
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