Patients Median Values Research Articles

Evaluation of serum MicroRNA 21, MicroRNA 192 and serum TGFβ1 in type 2 diabetes mellitus patients and their relation to diabetic nephropathy

BackgroundDiabetic nephropathy (DN) is a frequent and long-lasting microvascular consequence that has an established connection with diabetes. It serves as the primary etiological agent of end-stage renal disease, a critical renal disorder that develops on a worldwide level. The molecular pathophysiology of DN is multifactorial, such as transforming growth factor-beta [TGF-β] which affects the expression of miRNAs such as miRNA-21 and miRNA-192 during renal fibrosis. However, to date, the clinical application is inadequate due to discrepancies observed in the published data. This cross-sectional investigation aimed to assess the correlation between serum TGF-β1, miRNA-21 and 192, and glycemic control, metabolic abnormalities, and renal function in patients with type II diabetes.MethodsBased on the albumin/creatinine ratio (ACR), fifty subjects with type II diabetes were divided into three categories: Group I consisted of individuals with normoalbuminuria (n = 16), Group II of microalbuminuria (n = 16), and Group III of overt proteinuria (n = 18). All participants were subjected to the estimation of mature miRNA-21 and miRNA-192 by TaqMan two-step stem loop qRT-PCR and serum TGFβ1 level by ELISA.ResultsThere was an upregulation in miRNA-21 expression in the 3 different groups of patients (p value = 0.043). The serum fold change (FC) of miRNA-21 showed significantly greater median values in patients with overt proteinuria compared to those with normoalbuminuria (5.57 FC versus 1.11 FC, p = 0.017). A positive correlation (r = 0.343) (p = 0.013) was observed between the ACR and the median levels of miRNA-21, which was statistically significant. No statistically significant distinctions were detected in the concentrations of serum TGF-β1 or miRNA-192 among the three patient groups (p values of 0.234 and 0.225, respectively).ConclusionThe findings of the present research implied that miRNA-21 might function as an early indicator of renal pathology associated with diabetes mellitus (DM).

Linkage of cell division cycle 42 with clinical features, treatment response and survival in adult Philadelphia chromosome negative acute lymphoblastic leukemia

Cell division cycle 42 (CDC42) regulates the progression of leukemia via mediating proliferation and immune evasion of malignant cells. The study aimed to investigate the correlation of CDC42 with clinical features, treatment response, event-free survival (EFS) and overall survival (OS) in adult Philadelphia chromosome negative acute lymphoblastic leukemia (Ph− ALL) patients. CDC42 expression in bone marrow mononuclear cells was detected in 78 adult Ph− ALL patients and 10 donors using real-time reverse transcriptase-polymerase chain reaction. CDC42 was increased in adult Ph− ALL patients compared with donors (p < .001). Besides, elevated CDC42 was linked with pro-B ALL or early-T ALL (p = .038) and white blood cell (WBC) elevation at diagnosis (p = .025). Fifty (64.1%) and 23 (29.5%) patients had complete remission (CR) at 1 month and minimal residual disease (MRD) after CR, respectively. CDC42 was inversely associated with CR at 1 month (p = .034), but not MRD after CR (p = .066). Concerning survival, patients with CDC42 ≥ 3.310 (cut by median value in patients) showed a shortened EFS (p = .006) and OS (p = .036) compared to those with CDC42 < 3.310. In detail, patients with CDC42 ≥ 3.310 and CDC42 < 3.310 had 5-year EFS rate of 29.9% and 45.4%, and 5-year OS rate of 39.4% and 63.6%, correspondingly. Further multivariate Cox’s regression analyses revealed that CDC42 ≥ 3.310 was independently related to shorter EFS (hazard ratio = 2.933, p = .005). Elevated CDC42 is related with pro-B ALL or early-T ALL, WBC elevation at diagnosis, unfavorable treatment response and worse survival in adult Ph− ALL patients.

Identification of a new five-gene risk score for risk stratification and prognosis prediction in HCC

Approximately 80% of primary liver cancer (PLC) is hepatocellular carcinoma (HCC), and the prognosis of HCC patients is unfavorable. Further studies are required to develop new prognostic tools for predicting the HCC patients’ prognosis. The univariate Cox and LASSO regression were utilized to develop the multi-gene risk score. Single-sample Gene Set Enrichment Analysis (ssGSEA) was employed to assess differences of immune functions and cells. The model performance was evaluated by calibration curve and receiver operating characteristic curve (ROC). And qRT-PCR was utilized to evaluate the genes expression in clinical samples. Finally, a novel five-gene (KIF20A, CENPA, HMMR, G6PD, and ADH4) risk score was developed. Based on the median value of patients’ risk scores, patients were divided into two groups: high-risk group and low-risk group. The Overall survival (OS) of patients in high-risk group was obviously poorer than that in the low-risk group. And the five-gene risk score was an independent risk factor correlated with patients’ OS. Besides, a nomogram consisting of TNM stage and risk score was established. The results of decision curve, calibration curve, and ROC presented that the prognostic risk score and the nomogram had great predictive capability. Besides, ADH4’s mRNA was reduced in HCC tissues, while the mRNA of KIF20A, CENPA, HMMR, and G6PD were overexpressed in HCC tissues. We developed a novel five-gene risk score that could predict HCC patients’ prognosis. And these five genes could be promising therapeutic targets for HCC. The five-gene risk score and nomogram may be useful prognostic tools for HCC.

Modified Glasgow Prognostic Score, Prognostic Nutritional Index and ECOG Performance Score Predicts Survival Better than Sarcopenia, Cachexia and Some Inflammatory Indices in Metastatic Gastric Cancer

Background: Gastric carcinoma (GC) patients usually present with locally advanced or metastatic disease; therefore treatment aim is mainly palliation. In this study our purpose is to analyze the prognostic values of the sarcopenia index (SI), cachexia index (CIn) and other inflammatory indexes (advanced lung cancer inflammation index [ALI], modified Glasgow Prognostic Score [mGPS], prognostic index [PI], prognostic nutritional index [PNI] and neutrophil-to-lymphocyte ratio [NLR]) in metastatic GC patients.Methods: Data from the files of metastatic GC patients, who applied to Medical Oncology outpatient clinic in Marmara University Pendik Education and Research Hospital between January 2011 and June 2016, were retrospectively reviewed. Five hundred seventy patients with gastric cancer were detected. Exclusion criteria were the inability to reach the patient surveys for prognostic index calculations, the presence of additional comorbidities to affect the laboratory parameters, and the absence of metastatic disease. Finally, 87 of these patients were included in this study. For SI calculation L3 level muscle area was measured from patients’ computed tomography (CT) by a radiologist. SI reference value was obtained from western-EGWSOP (The European Working Group on Sarcopenia in Older People) and eastern (Harada Y, et al.) sources separately, as Turkey doesn’t have a reference value for SI. NLR cutoff value was accepted as the median value of patients’ NLR measurements. Statistical analysis was conducted using SPSS. Kaplan-Meier and Cox regression models were used to assess independent prognostic factors. The area under the curve was used to compare the prognostic value of indexes.Results: The median length of follow-up of 87 patients was nine months (1-64 mo,/s), and 78 patients died during follow-up. Fifty-nine patients were male (63%), and the median age was 62 (range, 23-88). According to univariate analysis high mGPS and PI score, PNI level <45, NLR level ≥ 3.41, ALI level <18, CI level under 35, SI (Harada Y, et al) ≤44.5 for males and ≤36.5 for females, ECOG score ≥ 2, weight loss more than 10% during last 6 mo, BMI under 24 were poor prognostic factors. Age, gender, having multiple organ metastasis, history of gastric surgery, positivity C-erb-B2, SI (EGWSOP) ≤52.4 for males, and ≤38.4 for females did not have any impact on survival. According to multivariate analysis, high mGPS (score 2) (HR 2,494, 95% CI 1.25–4 .94, p = 0.02), PNI (score 1) (HR 4.2, 95% CI 1.73–10.1, p < 0.001) and ECOG score (≥2) (HR 1.541, 95% CI 1,089-4,214, p = 0.004) have been found to be independent prognostic factors which are determining the survival. mGPS was found to be more valuable than other indexes for predicting mortality by measuring the AUC with ROC analysis.Conclusions: In our study, mGPS, PNI and ECOG score were independent indicators for shorter survival in metastatic gastric cancer patients. mGPS and PNI, which can be done by using only serum CRP, albumin level and complete blood count, might be inexpensive, practical and beneficial to use in routine clinical practice to determine survival.

Whose fault are wrong predictions: the clinician, the patient or the pigeon?

Whose fault are wrong predictions: the clinician, the patient or the pigeon?

Salivary MMP-9 in the detection of oral squamous cell carcinoma.

BackgroundOral squamous cell carcinoma (OSCC) is the most common malignant tumour of the oral cavity. Detection of OSCC is currently based on clinical oral examination combined with histopathological evaluation of a biopsy sample. Direct contact between saliva and the oral cancer makes measurement of salivary metalloproteinase- 9 (MMP-9) an attractive alternative.Material and MethodsIn total, 30 OSCC patients and 30 healthy controls were included in this prospective study. Saliva samples from both groups were collected, centrifuged and supernatant fluid was subjected to ELISA for assessment of MMP-9. The median salivary MMP-9 values with interquartile range (IQR) of OSCC patients and the control group were statistically analysed using the Mann-Whitney U-test. The receiver operating characteristic (ROC) curve was constructed and the area under curve (AUC) was computed.ResultsThe median absorbance MMP-9 value of the OSCC group was 0.186 (IQR=0.158) and that of control group was 0.156 (IQR=0.102). MMP-9 was significantly increased in the OSCC patients than in the controls by +19.2% (p=0.008). Median values in patients with recurrence and in patients with primary event were 0.233 (IQR=0.299) and 0.186 (IQR=0.134) respectively. MMP-9 was significantly increased in patients with primary event (p=0.017) compared to controls by +19.2%. No significant increase of MMP-9 level was detected when comparing patients with recurrence and healthy controls (+49.4%; p=0.074). The sensitivity value of MMP-9 was 100% whereas the specificity value was 26.7% with AUC of 0.698.ConclusionsThe present data indicates that the elevation of salivary levels of MMP-9 may be a useful adjunctive diagnostic tool for detection of OSCC. However, further studies are necessary to provide scientific and clinical validation. Key words:Oral squamous cell carcinoma, oral cancer, saliva, salivary diagnostics, cancer detection, MMP-9, metalloproteinases.

Clinical significance of serum Wisteria floribunda agglutinin positive Mac-2-binding protein level in non-alcoholic steatohepatitis.

To examine the relationship between the Wisteria floribunda agglutinin positive Mac-2-binding protein (WFA+ -M2BP) level and histological findings for patients with non-alcoholic steatohepatitis (NASH). A total of 134 NASH patients (mean age, 51.7years) were analyzed. We examined the effect of WFA+ -M2BP level on severity of liver fibrosis comparing with other laboratory markers, including aspartate aminotransferase (AST) to alanine aminotransferase (ALT) ratio, AST to platelet ratio index (APRI), FIB-4 index, platelet count and hyaluronic acid as serum liver fibrosis markers. Receiver-operator curve (ROC) analysis was performed for calculating the area under the ROC (AUROC). The WFA+ -M2B P-value ranged from 0.2 cut-off index (COI) to 9.6 COI (median, 0.9). The median values in each fibrosis stage were: 0.7 COI in F1, 0.7 COI in F2, 1.2 COI in F3 and 2.4 COI in F4 (P <0.001). For predicting liver cirrhosis (F4), WFA+ -M2BP level had the AUROC of 0.854 (sensitivity, 69.2%; specificity, 88.4%) and for predicting advanced liver fibrosis (≥F3), WFA+ -M2BP level yielded the second highest AUROC with a level of 0.842 (sensitivity, 73.7%; specificity, 80.2%) and for predicting significant liver fibrosis (≥F2), WFA+ -M2BP level yielded the highest AUROC with a level of 0.663 (sensitivity, 47.2%; specificity, 78.6%) among six liver fibrosis markers. The median values in patients with ballooning scores 1 (n = 58) and 2 (n = 76) were 0.6 and 1.1 COI, respectively (P <0.001). Serum WFA+ -M2BP level can be useful for assessing liver histological findings in patients with NASH.

CLINICAL VALUE OF CHROMOGRANIN A IN GASTROENTEROPANCREATIC NEUROENDOCRINE TUMORS

Background: Neuroendocrine tumors (NET) is a heterogeneous group of neoplasms characterized by hypersecretion of biologically active sub- stances that manifests by specific syndromes and determines the clinical course of the disease. The most common NET types are those of gastrointestinal tract. The obligatory biochemical marker used in the examination of NET patients is chromogranin A (CgA). Aim: Evaluation of the CgA value for diagnostics and monitoring of gastrointestinal NETs. Materials and methods: A comparative study of plasma CgA levels was performed in 146 patients with gastroenteropancreatic neuroendocrine tu- mors and 66 healthy individuals using the enzyme immunoassay “Chromogranin A ELISA kit” (Dako A/S, Denmark). Results: CgA levels were significantly higher in patients with NETs of all localizations, such as pancreas, stomach, gut, small and large bowel, than in the healthy subjects (р < 0.000001). In NET patients, CgA secretion was highly variable, with the highest value in the group of patients with gastric NETs (102000 U/l). The highest CgA medians were detected in patients with small intestinal (183.9 U/l), colon (148.4 U/l) and pancreatic (135.9 U/l) NETs. There was an association between CgA secretion and extension or activity of NETs, with the highest median values in patients with hepatic metastases (395 U/l) and those with carcinoid syndrome (352 U/l). The clinical significance of CgA as a NET marker was assessed using the cut-off value of 33 U/l, calculated according to the results in the control group. Overall diagnostic sensitivity of CgA in NET patients was high (85.8%) with a specificity of 98.5%. Conclusion: The results obtained confirm a high sensitivity of CgA as a NET marker whose determination helps to improve accuracy of diagnostics and to assess NET prevalence.

Abstract P5-19-25: Multi-institutional retrospective analysis of clinical and pathological factors predicting resistance to lapatinib-based therapy in HER2 positive metastatic breast cancer (HER2+ MBC)

Abstract Background The combination of the dual HER1/HER2 inhibitor lapatinib (L) and capecitabine (C) is a therapeutic option for patients (pts) with HER2+MBC whose disease progresses after treatment with the monoclonal antibody trastuzumab. At present time, no clinical or pathological factors except HER2 status are clearly recognized as predictors of the activity of LC. We conducted a retrospective analysis of pts with HER2-positive metastatic breast cancer receiving LC after trastuzumab failure to identify factors associated with resistance to LC. Materials and methods We collected clinical and pathological data from 151 pts with HER2+ MBC receiving LC after failing a prior trastuzumab-based treatment (either adjuvant or for metastatic disease) treated at 13 Italian Institutions between March 2007 and December 2013. Time to progression (TTP) and overall survival (OS), calculated by the Kaplan Meier (KM) method, were from LC treatment beginning to disease progression or to death in the absence of progression (TTP), and to the date of death or to the date of last follow-up (OS), respectively. LC resistance was defined as TTP from treatment initiation lower or equal to the median TTP for the overall population. KM curves were compared by the Log-rank test. Logistic regression analysis was used to study predictors of TTP below the median value for patients receiving LC. Analyses were performed using SPSS version 17.0 (SPSS Inc., Chicago, IL). Results At a median follow-up of 41 months (IQR 23-62), median TTP to LC therapy was 7 months (IQR 5.5-8.5) and median OS was 18 months (IQR 10-28). Fifteen pts were excluded because of short follow-up (i.e. on LC treatment and &amp;lt;7 months of fu). Of the remaining 136, a total of 74 pts with a PFS≤7 months were defined LC-resistant (LC-R) and a total of 62 pts were defined LC-sensitive (LC-S). All clinical and pathological variables analyzed resulted evenly distributed between the two groups, except best tumor response (CR+PR) to LC, which was higher in patients with LC-S disease (72% vs 29%, p&amp;lt;0.001). Conversely, best tumor response in LC-R patients showed higher rates of PD (43% vs 2%, p&amp;lt;0.001). Median OS was 14 months (IC 95% 11.4-22.6) and 26 months (IC 95% 22.5-29.5) in LC-R and LC-S pts, respectively (p&amp;lt;0.001). Although we could not find independent predictors of LC-R, factors indicating failure of the first-line trastuzumab based therapy, as PD as best tumor response and short duration of first-line trastuzumab, were associated to LC-R. Conclusions A short time to progression during capecitabine and lapatinib (LC-R) is associated with reduced OS in patients failing prior trastuzumab based therapy for HER2+ MBC. Patients who had modest clinical benefit from previous trastuzumab-based therapy could experience LC-R indicating the possibility of primary resistance to anti HER2-treatment. For these patients, alternative targeting strategies are urgently needed. Citation Format: Stefania Gori, Valentina Rossi, Monica Turazza, Elena Fiorio, Alessandra Fabi, Giancarlo Bisagni, Jennifer Foglietta, Daniele Santini, Ida Pavese, Alberto Zambelli, Patrizia Vici, Vita Leonardi, Sandro Barni, Silvana Saracchini, Giuseppe Bogina, Simona Duranti, Alessandro Inno, Gianluigi Lunardi, Filippo Montemurro. Multi-institutional retrospective analysis of clinical and pathological factors predicting resistance to lapatinib-based therapy in HER2 positive metastatic breast cancer (HER2+ MBC) [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P5-19-25.

Vascular endothelial growth factor serum levels in children with newly diagnosed rhabdomyosarcoma

The adverse prognostic impact of elevated levels of circulating Vascular Endothelial Growth Factor (VEGF) is described in several malignancies. However, no information is available in childhood rhabdomyosarcoma (RMS). In the present study, serum VEGF-A (sVEGF-A) was measured at diagnosis in a series of patients with RMS. sVEGF-A was assessed retrospectively in 17 newly diagnosed RMS patients. sVEGF-A concentrations were determined by quantitative enzyme-linked immunoabsorbent ELISA kit and their possible associations with age at diagnosis, gender, histology, primary site, primary size, Intergroup Rhabdomyosarcoma Study (IRS) post-surgical group, and outcome were investigated. sVEGF-A median value in patients with RMS was significantly higher than in controls: 499.0 pg/ml, range: 2,648.0 versus 301.5 pg/ml, range: 716.0 (P = 0.013). Although not statistically significant probably due to the limited number of patients, sVEGF-A median levels resulted higher in unfavorable primary sites (277.0 vs. 539.0 pg/ml; P = 0.31), and advanced groups (390.0 vs. 715.0; P = 0.29). Patients with shorter 5-year overall survival (OS) and 5-year progression-free survival (PFS) times also had higher sVEGF-A levels, although again the difference was not statistically significant (P = 0.18 and P = 0.22, respectively). Circulating VEGF is significantly increased in pediatric patients with newly diagnosed RMS. Further studies in larger series of RMS patients are needed to understand whether measurements of circulating VEGF might have a role in assessing prognosis and modulating treatment.

Up-regulation of the Extrinsic Coagulation Pathway in Acute Asthma—A Case Study

Introduction. In the normal airway, the hemostatic balance is antithrombotic and favors fibrinolysis. Acute asthma is associated with inflammatory cell infiltrate and plasma exudation in the airways. Postmortem specimens following status asthmaticus suggest a role for the activation of the extrinsic coagulation cascade and intraluminal fibrin formation. The authors report a chance observation of fibrin formation in the airways of a patient with moderate asthma 5 days before a severe exacerbation requiring hospital admission. Methods. Alpha-2 macroglobulin, an index of plasma leakage, coagulation factors, and D-dimers were measured by enzyme-linked immunosorbent assay (ELISA) in hypertonic saline-induced sputum, as part of a study into airway repair in stable asthma. All subjects were required to have stable symptoms and measures of asthma prior to sampling. Results. The subject's baseline forced expiratory volume in one second (FEV1) was 94% predicted and fraction of exhaled nitric oxide (FeNO) level was 30 ppb prior to sputum induction. Differential sputum cell count revealed an airways neutrophilia (neutrophils 81.1%, eosinophils 0.19%). D-dimers were 70-fold and 22-fold higher than the median value for patients with stable moderate and severe asthma, respectively. Plasma exudation was 42-fold higher than in stable moderate asthma, but on a par with levels found in severe stable asthma, and locally produced coagulation factors may therefore be involved. Levels of fibrinogen, plasminogen, plasminogen activator inhibitor (PAI)-1 and thrombin-activatable fibrinolysis inhibitor (TAFI) were all at least an order of magnitude higher than those seen in stable moderate or severe asthma. Conclusions. Acute exacerbation of moderate asthma appears to be associated with a shift to a profibrinogenic, possibly antifibrinolytic, environment in the airways.

Development of an enzyme-linked immunosorbent assay for the detection of human calretinin in plasma and serum of mesothelioma patients

BackgroundCalretinin is one of the well-established immunohistochemical markers in the diagnostics of malignant mesothelioma (MM). Its utility as a diagnostic tool in human blood, however, is scarcely investigated. The aim of this study was to develop an enzyme-linked immunosorbent assay (ELISA) for human calretinin in blood and to assess its usefulness as a potential minimally invasive diagnostic marker for MM.MethodsInitially, attempts were made to establish an assay using commercially available antibodies and to optimize it by including a biotin-streptavidin complex into the assay protocol. Subsequently, a novel ELISA based on polyclonal antibodies raised in rabbit immunized with human recombinant calretinin was developed. The assay performance in human serum and plasma (EDTA/heparin) and the influence of calcium concentrations on antibody recognition were studied. Stability of spiked-in calretinin in EDTA plasma under different storage conditions was also examined. In preliminary studies serum and plasma samples from 97 healthy volunteers, 35 asbestos-exposed workers, and 42 MM patients were analyzed.ResultsThe mean detection range of the new ELISA was 0.12 to 8.97 ng/ml calretinin. The assay demonstrated markedly lower background and significantly higher sensitivity compared to the initially contrived assay that used commercial antibodies. Recovery rate experiments confirmed dependence of calretinin antibody recognition on calcium concentration. Calcium adjustment is necessary for calretinin measurement in EDTA plasma. Spiked-in calretinin revealed high stability in EDTA plasma when stored at room temperature, 4°C, or after repeated freeze/thaw cycles. Median calretinin values in healthy volunteers, asbestos workers, and MM patients were 0.20, 0.33, and 0.84 ng/ml, respectively (p < 0.0001 for healthy vs. MM, p = 0.0036 for healthy vs. asbestos-exposed, p < 0.0001 for asbestos-exposed vs. MM). Median values in patients with epithelioid and biphasic MM were similar. No influence of age, gender, smoking status, or type of medium (plasma/serum) on calretinin values was found.ConclusionsThe novel assay is highly sensitive and applicable to human serum and plasma. Calretinin appears to be a promising marker for the blood-based detection of MM and might complement other markers. However, further studies are required to prove its usefulness in the diagnosis of MM patients.

Adiponectin is associated with abnormal lipid profile and coronary microvascular dysfunction in patients with dilated cardiomyopathy without overt heart failure

Reduced plasma adiponectin has been associated with abnormal lipid profile, reduced left ventricle (LV) function, and the extent of coronary atherosclerosis in coronary artery disease. The aim of this study was to assess these relationships in patients with dilated cardiomyopathy (DCM) without overt heart failure. Plasma adiponectin was measured in 55 DCM patients (age, 59 ± 12 years; male, 36; body mass index [BMI], 26.9 ± 0.49 kg/m2; LV ejection fraction, 39.8% ± 1.3%; New York Heart Association class I-II) and in 40 age- and BMI-matched healthy controls. In a subset of 25 patients, myocardial blood flow (MBF) was measured at rest and during intravenous dipyridamole (0.56 mg/kg in 4 minutes) by positron emission tomography and 13N-ammonia as a flow tracer. Adiponectin was 6.6 ± 0.34 μg/mL in controls and 10.9 ± 0.85 μg/mL in DCM patients (P < .001), where it was related inversely with BMI (P = .009) and directly with brain natriuretic peptide (P = .017), high-density lipoprotein (HDL) cholesterol (P = .002), and MBF dipyridamole (P = .020). Adiponectin lesser than median value in patients was associated with higher total to HDL cholesterol ratio (4.8 ± 0.24 vs 3.9 ± 0.18, P = .009) and lower MBF reserve (1.76 ± 0.16 vs 2.43 ± 0.19, P = .01). These results could suggest that down-regulation of the adiponectin levels and reduced HDL cholesterol have a key role in causing impaired coronary function and myocardial perfusion in DCM.

Abstract 5502: Adiponectin Mediates the Effects of Atherogenic Risk Profile on the Coronary Microcirculation in Patients with Idiopathic Lv Systolic Dysfunction

Purpose. Adiponectin (ADN), a biologically active protein produced by the adipose tissue, has protective vascular effects. Accordingly, ADN plasma levels are reduced in patients with coronary artery disease (CAD) while in heart failure ADN tends to increase. We hypothesized that ADN plasma levels could mediate the effects of atherogenic risk profile on the coronary microcirculation of patients with early LV systolic dysfunction (ILVDys) not secondary to established CAD. Methods. Plasma ADN was measured in 55 patients (age 59±1 yrs, 36 males, BMI 26.9±0.49 Kg/m 2 , mean±sem) with angiographically normal coronary arteries, LV systolic dysfunction (LVEF 39.8±1.3 % range 22–54 %) but without overt heart failure (NYHA class I–II) and in 40 age- and BMI-matched healthy controls by using a specific Elisa (Linco Res). BMI, cholesterol and glucose profiles were assessed in all. In a subset of 25 patients coronary microvascular function was studied by PET and 13 N-Ammonia as a flow tracer. Myocardial blood flow (MBF) was measured at rest and during i.v. dipyridamole (Dip) (0.56 mg/Kg in 4 min). Results. ADN was 6.6±0.34 μg/ml in controls and 10.9±0.85 in ILVDys patients (p&lt;0.001). In patients ADN levels were inversely related with BMI (p=0.009) and directly related with age (p=0.007), HDL Cholesterol (p=0.003) and MBF Dip (0.020). Patients showing more severe coronary microvascular dysfunction (MBF Dip&lt;1.42 ml/min/g, median value in patients) had significantly depressed ADN (9.7±2.3 vs 13.7±1.6, p=0.021) as compared with the remaining patients. Conclusions. This is the first study which associates adiponectin plasma levels with atherogenic risk profile and coronary microvascular function in patients with idiopathic LV dysfunction. These results suggest that adiponectin signal is strongly involved in mediating coronary vascular function independently of the presence of overt CAD or heart failure.

Dysfunction of Toll-Like Receptor-2, −4, and −6: Is There a Role in the Pathogenesis of Immunodeficiency in Multiple Myeloma?.

Infections are a major cause of death in multiple myeloma (MM). There is limited information for the possible role of innate immunity in the pathogenesis of immunodeficiency in MM. Innate immune detection of pathogens relies on specific classes of microbial sensors, such as Toll-like receptors (TLRs) that detect structural patterns, which do not exist in the host. The aim of this study was to evaluate the expression and function of TLRs in newly diagnosed patients with MM. According to our knowledge, such information is not available in the literature. Thirty-two MM patients at diagnosis (18M/14F; median age 69 years), 6 MGUS patients and 14 healthy, age- and gender-matched controls were studied. Seven patients had stage 1, 14 stage 2 and 11 stage 3 myeloma, according to ISS. After the collection of peripheral blood, mononuclear cells (PBMCs) were isolated by Ficoll centrifugation (Histopaque-1077; Sigma-Aldrich). These cells were measured for the expression of TLRs (antibodies from eBioscience) using fluorescence activated flow cytometry (FC 500, Beckman Coulter). In addition, 1x106 cells/ml were cultured in 5% FCS 1% pen/strep RPMI in the presence or absence of various TLR ligands and supernatants collected after 20h. These were examined for the presence of inflammatory cytokines (tumor necrosis-alpha, TNF-α; and interleukin-6, IL-6) by ELISA (Becton Dickinson). We found that although patients with MM express TLRs in PBMCs, their response to certain TLR ligands is defective when compared to healthy controls. TLR2, TLR4 and TLR6 of PBMCs of healthy controls reacted to the presence of their respective ligands PAM3CYS (gram positive and negative bacterial), LPS (gram negative) and FLT-1 (gram positive) producing TNF-α at a median value of 2.4 ng/mL (range: 1–3 ng/ml), while their action in patients with MM was significantly reduced (median value of TNF-α: 190 pg/ml; range 100–500 pg/ml; p<0.001). The reduced response of TLR4 in MM patients was independent of the LPS concentration used. On the contrary, TLR7 and TLR8 from MM reacted normally to their ligand R-848 (Imiquimob) to secrete high levels of TNF-α (median value for patients and controls: 4.5 and 4.2 ng/ml, respectively; p=NS). NOD1, another pattern recognition receptor that recognizes bacterial peptidoglycans also reacted normally in MM. Similar observations have been made for IL-6 expression. There was no difference in terms of TLRs function between MGUS patients and controls and between myeloma patients of different disease stages. In seven MM patients, cells were pre-treated with bortezomib for 30 min before addition of the TLR ligand. Bortezomib administration abrogated TLR 7/8 response in PBMCs even at very low concentrations (1nM). This study suggests that there is a significant defect in TLR function in MM, especially of these involved in immunity against bacterial infections. Thus the immune system fails to receive early priming signal, which may contribute to the increased rate of infections observed in MM. The restoration of function of TLRs to their normal levels has the potential to improve bacterial immunity in MM.

Calprotectin, a faecal marker of organic gastrointestinal abnormality

Calprotectin may therefore exert its effects by reducing local concentrations of zinc, which indicates that it may be involved in the regulation of inflammatory reactions by its inhibition of several zinc-dependent metalloproteinases. The protein is remarkably resistant to degradation in vivo and in vitro in the presence of calcium, 5,6 so faecal samples can be sent to the laboratory by post. The upper reference limit in faeces is 10 mg/L, but with a recent refinement of the assay, which does away with the messy step of homogenisation and which increases yields, the limit is 50 mg/L. 7 Several groups have confirmed the observation 5 that calprotectin can be assayed in simple buffer extracts of small (5 g) faecal samples, and that increased concentrations (>10 mg/L) are found in most patients with inflammatory bowel disease (IBD) or gastric or colorectal cancer (panel). It was therefore quite clear at an early stage that the faecal calprotectin test was nonspecific for type of organic bowel disease. In different studies the median values in patients with colorectal cancer ranged from 33 to 214 mg/L. In one study 23 patients were also tested after resection, and the median dropped from 75 to 9 mg/L. 8

Assessment of the neutrophil dominating protein calprotectin in feces. A methodologic study.

This study describes methods for extraction and quantification of calprotectin (L1 protein) in feces by enzyme immunoassay. This protein is a prominent antimicrobial component of neutrophils, monocytes, macrophages, and squamous epithelia. Calprotectin was stable in feces during storage for 7 days at room temperature. Small fecal samples taken from a 24-h feces collection gave a reliable estimate of calprotectin. Within-assay precision was 1.9%, and between-assay precision 14.8%. In healthy subjects (n = 33) median fecal calprotectin was 2025 micrograms/l and in hospital controls (n = 40) 10,500 micrograms/l. Median values in patients with Crohn's disease (n = 21) was 43,000 micrograms/l and in ulcerative colitis (n = 17) 40,000 micrograms/l. Fecal calprotectin was significantly correlated to fecal alpha 1-antitrypsin in the patients with Crohn's disease. Ten of 11 patients with gastrointestinal carcinomas had calprotectin level above the suggested reference limit of 6740 micrograms/l.