The importance microRNAs as a biomarker in lung cancer.

INTRODUCTION: Lung cancer (LC) is the most common cancer  in the world.Well known  causes are  long term  smoking, environmental influences and genetic variations. LC  is divided into two main types based on their histological phenotypes; small cell lung cancer (SCLC), and non-small cell lung cancer (NSCLC). The high specificity of these new screening methods, which are non-invasive, safe, inexpensive and simple to perform, is important in the early diagnosis and prognosis of cancer. MicroRNAs are  significant biomarkers on the diagnosis metastasis and targeted therapies of NSCLC. In our study, we aimed to investigate the potential of using microRNAs as a biomarker in the early diagnosis of lung cancer. MATERIAL AND METHOD: Twenty patients diagnosed with lung cancer and  twenty healthy individuals of the same age and gender were selected as the control group.  Sixteen microRNAs were studied from blood samples. RESULT: Sixteen miRNAs (Let -7c, Let-7g, miR-1, miR-21, miR-29a, miR-31, miR-34a, miR 103a, miR-141, miR-155, miR-193b, miR-200b, miR-205, miR-340, miR-486, miR-708) were selected for tests and MiR 181 and miR 192 were used as the endogenous control group in line with their binding potentials and gene expression levels. The most specific and sensitive miRNAs were mirR-29a, miR-103a, and miR486 according to endogen controls in patients and healthy subjects. DISCUSSION: A meta-analysis study showed that circulating miRNAs could be promising biomarkers for early diagnosis of lung cancer. Overall, 17 studies were included evaluating 35 miRNA markers and 19 miRNA panels in serum or plasma. The potential role of circulating miRNAs for non-invasive lung screening has been highlighted. In conclusion, there is a need for further validation studies for the use of three  miRNAs as a biomarker in the early diagnosis and prognosis of lung cancer.

Use of MicroRNAs as biomarkers in the early diagnosis of prostate cancer.

INTRODUCTION: Prostate cancer (PCa) is a common type of cancer in western countries and prominent cause of mortality in men. The aim of the study was to analyze circulating miRNAs as biomarkers in the sera of healthy individuals and prostate cancer cases without biopsy. MATERIAL AND METHODS: Twenty prostate cases, age (mean and range) 61,4±12.1 (45-73), and twenty healthy men, age 59,3±11.2 (44-70) were included to the study. The mean and range of prostate spesific antigen (PSA) in cancer cases and healthy individuals were 6.79±2.84 ng/ml (2.25-14.7) and 3.8±2.2 ng/ml (1.3-7.8) respectively. RESULTS: Seven miRNAs including two internal controls (Let7c, miR125b, miR141, miR145, miR 155, miR181 ve miR192) were evaluated in two groups. The level of miR141 was significantly lower in PCa cases than healthy individuals (p=0,004), and miR155 was significantly higher (p=0,005) in PCa cases. Both miRNAs were explored sensitive and spesific in the ROC analysis. Tumor mass were found to be associated with the level of miR-125b and miR-145. Conclusion; validation studies are required in wider patient groups in the subject of tumor effect and miRNA biomarkers in prostate cancer.

MicroRNAs as biomarkers for breast cancer.

Breast cancer is the most common type of cancer among women and the most frequent cause of death due to cancer among women. The lack of standard biomarkers in the early diagnosis of breast cancer, microRNAs (miRNA) have been of interest recently. Although, miRNAs are 19-24 nucleotide-long non-coding RNA species, they have crucial roles in many areas from organogenesis to carcinogenesis. This study has been conducted to investigate miR 21, miR 27b, miR 125a, miR 155, miR 200c, miR 335 miR373 as biomarkers in the early diagnosis of breast cancer; a selection based on the literature. Two miRNAs, miR 181 and miR 192 were selected as the endogenous control. MiRNAs were obtained from 5 cc blood samples taken from 20 breast cancer patients and 20 healthy people. 10 microRNAs were studied using Real Time PCR method. As a result, the quantities of miR 21, miR155 and miR125 ​​were significantly higher in the breast cancer group than in healthy controls. We suggest that performing validation studies in wider populations can help the use of miRNAs as biomarkers in the early diagnosis of breast cancer.

A Concise Review on the Frequency, Major Risk Factors and Surveillance of Hepatocellular Carcinoma (HCC) in ß-Thalassemias: Past, Present and Future Perspectives and the ICET-A Experience.

Due to the recent alarming increase in the incidence of hepatocellular carcinoma (HCC) in thalassemias, the present report reviews briefly the frequency, the major risk factors, and the surveillance of HCC in ß-thalassemias. Over the past 33 years, 153 cases of HCC were reported in patients with thalassemia, mainly in Italy and Greece. Among HCV-infected patients, additional factors promoting the development of HCC included: advanced age, male sex, chronic hepatitis B (CHB) co-infection, and iron overload. For early diagnosis of HCC, sequential ultrasound screening is recommended especially for thalassemia patients with chronic hepatitis C (CHC), which coincides with (one or more) additional risk factors for HCC. Here we report also the preliminary data from thalassemic patients, above the age of 30 years, followed in 13 ICET-A centers. The total number of enrolled patients was 1,327 (males: 624 and 703 females). The prevalence of HCC in thalassemia major patients [characterized by transfusion-dependency (TDT)] and thalassemia intermedia [characterized by nontransfusion dependency (NTDT)] was 1.66 % and 1.96 %, respectively. The lowest age at diagnosis of HCC was 36 years for TDT and 47 years for NTDT patients. We hope that this review can be used to develop more refined and prospective analyses of HCC magnitude and risk in patients with thalassemia and to define specific international guidelines to support clinicians for early diagnosis and treatment of HCC in thalassemic patients.

Thyroid Disorders in Homozygous ß-Thalassemia: Current Knowledge, Emerging Issues and Open Problems.

Changes in thyroid function and thyroid function tests occur in patients with ß-thalassemia major (TM). The frequency of hypothyroidism in TM patients ranges from 4% to 29 % in different reports. The wide variation has been attributed to several factors such as patients' genotype, age, ethnic heterogeneity, treatment protocols of transfusions and chelation, and varying compliance to treatment. Hypothyroidism is the result of primary gland failure or insufficient thyroid gland stimulation by the hypothalamus or pituitary gland. The main laboratory parameters of thyroid function are the assessments of serum thyroid-stimulating hor-mone (TSH) and serum free thyroxine (FT4). It is of primary importance to interpret these measurements within the context of the laboratory-specific normative range for each test. An elevated serum TSH level with a standard range of serum FT4 level is consistent with subclinical hypothyroidism. A low serum FT4 level with a low, or inappropriately normal, serum TSH level is consistent with secondary hypothyroidism. Doctors caring for TM patients most commonly encounter subjects with subclinical primary hypothyroidism in the second decade of life. Several aspects remain to be elucidated as the frequency of thyroid cancer and the possible existence of a relationship between thyroid dysfunction, on one hand, cardiovascular diseases, components of metabolic syndrome (insulin resistance) and hypercoagulable state, on the other hand. Further studies are needed to explain these emerging issues. Following a brief description of thyroid hormone regulation, production and actions, this article is conceptually divided into two parts; the first reports the spectrum of thyroid disease occurring in patients with TM, and the second part focuses on the emerging issues and the open problems in TM patients with thyroid disorders.

An ICET-A survey on occult and emerging endocrine complications in patients with ß-thalassemia major: Conclusions and recommendations.

In adult thalassemia major (TM) patients, a number of occult and emerging endocrine complications, such as: central hypothyroidism (CH), thyroid cancer, latent hypocortisolism, and growth hormone deficiency (GHD) have emerged and been reported. As the early detection of these complications is essential for appropriate treatment and follow-up, the International Network of Clinicians for Endocrinopathies in Thalassemia and Adolescent Medicine (ICET-A) promoted a survey on these complications in adult TM patients, among physicians (pediatricians, hematologists and endocrinologists) caring for TM patients in different countries. The data reported by 15 countries are presented.The commonest endocrine complications registered in 3.114 TM adults are CH and GHD (4.6 % and 3.0 %, respectively), followed by latent hypocortisolism (1.2%). In 13 patients (0.41%) a cytological papillary or follicular thyroid carcinoma was diagnosed in 11 and 2 patients, respectively, and a lobectomy or thyroidectomy was carried out. Of 202 TM patients below the age of 18 years, the  reported endocrine complications were: GHD in 4.5%, latent hypocortisolism in 4.4% and central hypothyrodisim in 0.5%. Transition phase was an area of interest for many clinicians, especially as patients with complex chronic health conditions are responding to new treatments extending their lifespan beyond imagination.. In conclusion, our survey provides a better understanding of  physicians' current clinical practices and beliefs in the detection, prevention and treatment of some endocrine complications prevailing in adult TM patients. Regular surveillance, early diagnosis, treatment and follow-up in a multi-disciplinary specialized setting are recommended.

Report on Ten Years' Experience of Premarital Hemoglobinopathy Screening at a Center in Antalya, Southern Turkey.

Thalassemia and hemoglobinopathies are a major public health problem in Turkey. Hemoglobinopathy prevention programs (HPPs) were started in 33 provinces situated in Thrace, Marmara, Aegean, Mediterranean and South Eastern regions of Turkey in 2003. A premarital hemoglobinopathy test is mandatory and free of charge in this program. According to the Ministry of Health reports, 46 first level hemoglobinopathy diagnostic centers were established for premarital tests. Within the last 10 years, approximately 79.0% of married individuals per year were screened by the centers. While the percentage of premarital screening tests was 30.0% of all couples in 2003, it reached 86.0% in 2013. The number of newborn with thalassemia and hemoglobinopathies were 272 in 2002 and dropped to 25 in 2013. There has been a 90.0% reduction in affected births. Our hemoglobinopathy diagnostic center was established in 2003 and licensed by the Ministry of Health in 2004. We studied a total of 89,981 blood samples from premarital tests for 10 years and the incidence of ß- and α-thalassemia (ß- and α-thal) trait was found to be 6.57 and 3.56%, respectively. The distribution of the most common abnormal hemoglobins (Hbs) was: Hb S (HBB: c.20A > T) (0.31%), Hb D-Los Angeles (HBB: c.364G > C) (0.15%), Hb G-Coushatta (HBB: c.68A > C) (0.06%) and Hb E (HBB: c.79G > A) (0.02%). A total of 60 couples, both carrying ß-thal trait, were directed to the prenatal diagnosis (PND) center in 10 years. The premarital hemoglobinopathy screening program is running successfully at our center and other centers in Turkey.

The protective effect of Capparis ovata on 6-mercaptopurine-induced hepatotoxicity and oxidative stress in rats.

Capparis ovata is a member of Capparidacaeae family has been used in phytomedicine with a lot of positive effects such as an antioxidative, antihyperlipidemic, anti-inflammatory, and antihepatotoxic agent. The aim of this study was to research the protective effect of C. ovata on 6-mercaptopurine (6-MP) induced to hepatotoxicity and oxidative stress in rats. The rats were divided into 4 groups: control, 6-MP, C. ovataovate, and 6-MP + C. ovata. A complete blood count was performed, liver function test and antioxidant enzymes levels such as superoxide dismutase, glutathione peroxidase, catalase, and malondialdehyde were measured in blood before and after a 14-day test period. White blood cell and platelet counts were lower in the 6-MP group than other 3 groups (P < 0.005). Hepatic transaminase levels were higher in 6-MP group than the 3 groups (P < 0.05). Superoxide dismutase, glutathione peroxidase, and CAT levels were lower and malondialdehyde was higher in blood samples in 6-MP group than other 3 groups (P < 0.005). In conclusion, our tests were showed that C. ovata may be useful in patients receiving 6-MP therapy to prevent hepatotoxicity and in order to maintain uninterrupted therapy possibly reducing the risk of relapse. Although additional studies ensure that Capparis does not affect 6-MP antileukemic activity. We believe these results are important contribution to the literature.

The Thalassemia center of Antalya State Hospital: 15 years of experience (1994 to 2008).

The Thalassemia center of Antalya State Hospital was established in 1994 in Antalya, Turkey. The number of newborns with thalassemia decreased statistically because of actions taken as a result of prevention studies. A total of 388 patients, including 246 with thalassemia major (63.4%), 86 with thalassemia intermediate (22.1%), 23 with sickle cell+ß-thalassemia (5.9%), 20 with sickle cell disease (5.1%), and 13 with other hemoglobin abnormalities (3.3%), were studied. Complications were found to be as follows: cardiomyopathy in 45 of them (11.5%), diabetes mellitus in 10 (2.5%), hypothyroidism in 17 (4.3%), hypoparathyroidism in 2 (0.5%), osteoporosis in 53 (13.6%), growth retardation in 110 (28.3%), and hypogonadism in 75 patients (19.3%). The incidence of autoantibody and alloantibody in patients with thalassemia major was 5.6% and 10.5%, respectively. Transfusion-transmission diseases evaluated in patients found the incidence of hepatit A virus IgG to be 97.5%, that of HBs Ab to be 99.5%, HBs Ag to be 0.5%, HCV Ab to be 18%, CMV IgG to be 72.3%, CMV IgM to be 2%, and HIV-Ab to be 0%. Therefore, for the first time in our country the thalassemia center was established as a model and included subunits and a team. It served all patients for 15 years until the team was dispersed; thereafter, most of the patients were not followed up on a regular basis.

Iron and ferritin levels in saliva of patients with thalassemia and iron deficiency anemia.

Most of the techniques for measuring iron stores such as serum iron concentration, iron binding capacity, serum ferritin level, liver biopsy can be troublesome or invasive for patients with thalassemia. The salivary iron measurement could be of potential advantage being an easy and non invasive approach for diagnosis of iron deficiency and iron overload . The aim of this study was to compare the levels of iron and ferritin in saliva and serum of patients affected by thalassemia or iron deficiency anemia. For this purpose, 96 patients with iron overload (71 with thalassemia major, 10 with thalassemia intermedia and 15 with thalassemia trait), 30 patients with iron deficiency anemia, and 35 healthy children as control group were involved in this study. Their saliva and serum iron and ferritin levels were measured. Iron and ferritin levels were higher in iron overload groups than in control group and lower in iron deficiency group (p<0.05). Furthermore serum and saliva iron and ferritin levels paralleled in all groups. In conclusion, iron and ferritin saliva can be routinely used for diagnosis of both iron overload and deficiency; furthermore this procedure may be an important advantage for blood donors being easily available and not invasive.

Iron chelation with deferasirox in adult and pediatric patients with thalassemia major: efficacy and safety during 5 years' follow-up.

Patients with ß-thalassemia require lifelong iron chelation therapy from early childhood to prevent complications associated with transfusional iron overload. To evaluate long-term efficacy and safety of once-daily oral iron chelation with deferasirox, patients aged ≥ 2 years who completed a 1-year, phase 3, randomized trial entered a 4-year extension study, either continuing on deferasirox (deferasirox cohort) or switching from deferoxamine to deferasirox (crossover cohort). Of 555 patients who received ≥ 1 deferasirox dose, 66.8% completed the study; 43 patients (7.7%) discontinued because of adverse events. In patients with ≥ 4 years' deferasirox exposure who had liver biopsy, mean liver iron concentration significantly decreased by 7.8 ± 11.2 mg Fe/g dry weight (dw; n = 103; P < .001) and 3.1 ± 7.9 mg Fe/g dw (n = 68; P < .001) in the deferasirox and crossover cohorts, respectively. Median serum ferritin significantly decreased by 706 ng/mL (n = 196; P < .001) and 371 ng/mL (n = 147; P < .001), respectively, after ≥ 4 years' exposure. Investigator-assessed, drug-related adverse events, including increased blood creatinine (11.2%), abdominal pain (9.0%), and nausea (7.4%), were generally mild to moderate, transient, and reduced in frequency over time. No adverse effect was observed on pediatric growth or adolescent sexual development. This first prospective study of long-term deferasirox use in pediatric and adult patients with ß-thalassemia suggests treatment for ≤ 5 years is generally well tolerated and effectively reduces iron burden. This trial was registered at www.clinicaltrials.gov as #NCT00171210.

The investigation of gingival iron accumulation in thalassemia major patients.

BACKGROUND: Thalassemia major (TM) is an autosomal-recessive genetic blood disorder. Regular blood transfusions to improve chronic anemia caused by ineffective erythropoiesis and hemolysis lead to iron overload in many organs in TM patients. The aim of this study was to investigate the periodontal status and the iron accumulation in gingival tissues of TM patients and assess whether iron deposition in gingival biopsies could be an alternative method for the diagnosis of body iron overload in TM patients. MATERIALS AND METHODS: This study was conducted on 22 TM patients and 20 healthy matched controls. Plaque index, gingival index, and probing pocket depth were measured and gingival biopsies were obtained in all subjects. Venous blood samplings and liver biopsies were carried out only in patients with thalassemia. Gingiva and liver tissue samples were evaluated histopathologically for inflammation, iron accumulation, and fibrosis. RESULTS: There was no difference between the groups regarding periodontal health, and all patients had mild gingivitis. Gingival iron accumulation was observed only in the TM group. The iron accumulation was detected in the liver of all the patients with thalassemia. The gingival iron accumulation was correlated with neither serum ferritin levels nor hepatic iron accumulations. CONCLUSIONS: The periodontal tissues are affected by iron accumulation as well as hepatic, cardiac, and endocrine tissues in TM patients. Further studies investigating the usage of the gingival biopsy for prediagnosis of body iron overload in TM patients are needed.

Evaluation of angiogenesis with vascular endothelial growth factor in patients with thalassemia major.

BACKGROUND: Thalassemia major (TM) is an important cause of severe anemia that necessitates regular blood transfusion to prevent the profound weakness and cardiac decompensation caused by the anemia. However, iron overloading is an inevitable consequence of prolonged transfusion therapy. In addition, extramedullary hematopoiesis and hemosiderosis cause spleen, liver and marrow enlargement. In recent years the role of angiogenesis has been investigated in physiological and pathological conditions. However, it is known that angiogenetic factors, especially the vascular endothelial growth factor (VEGF), cause differentiation of the hemangioblast. METHODS: The effect of angiogenesis hasn't been investigated in TM patients yet, and in this study, angiogenesis was researched in 43 thalassemic patients by serum VEGF measurement. RESULTS: VEGF levels were not affected by hemoglobin levels, ferritin levels, or chelation type (P > 0.05). However, VEGF was positively affected by chelation starting age and negatively affected by yearly transfusion requirement of TM patients (P < 0.05). In addition, VEGF of patients who underwent splenectomy were higher than those who didn't undergo splenectomy (P < 0.05). CONCLUSION: Early chelating age will negatively influence the VEGF level, which increases angiogenesis, however, early starting transfusion age and regular blood transfusion will positively influence the VEGF level, which decreases angiogenesis in thalassemic patients.

Modulation of redox pathways in neutrophils from sickle cell disease patients.

OBJECTIVE: Interaction of nitric oxide (NO) with enzymatic sources of reactive species exerts modulatory actions on inflammatory signaling mechanisms. MATERIALS AND METHODS: NADPH oxidase, total peroxidase, cyclooxygenase (COX) activity, and NO consumption were measured in neutrophils isolated from sickle cell disease (SCD) patients and healthy controls. Glutathione (GSH) levels and expression of inducible NO synthase (NOS-2) were also analyzed to assess intracellular redox state and NO production, respectively. RESULTS: Functional assay of NADPH oxidase was performed by measuring superoxide release, which was similar in control and SCD, both at basal conditions and in response to N-formyl-methionyl-leucyl-phenylalanine stimulation. Peroxidase activity, assessed spectrophotometrically, was not significantly different in SCD neutrophils compared to controls. Total COX activity, measured via an assay kit, was significantly increased in SCD neutrophils. The increase in total COX activity observed in SCD was due to enhanced activity of COX-2, differentiated by using the isoform-specific inhibitors DuP-697 and SC-560. Western blot analysis of COX-2 protein in SCD and control neutrophils confirmed increased enzyme activity in the diseased group. Western blot analysis of neutrophil lysates from SCD patients showed significantly increased NOS-2 protein content, compared to controls. Spectrophotometric measurement of GSH and nitrate/nitrite levels showed a decrease in GSH and an increase in nitrate/nitrite content in SCD neutrophils. Electrochemical measurement of NO consumption both under basal conditions and after N-formyl-methionyl-leucyl-phenylalanine stimulation revealed a significant decrease in SCD neutrophils compared to controls. CONCLUSIONS: Depletion of GSH in SCD neutrophils may impact on rates of NO consumption and reflects increased oxidative stress associated with neutrophil activation.

Does the periodontal health of thalassemia major patients have an impact on the blood lipid profiles? A preliminary report.

Cardiac symptoms and premature death from cardiac causes are still suggested to be a major problem in thalassemic patients. One of the main reasons for the cardiovascular events in thalassemia major (TM) patients has suggested having iron overload, in addition to other reasons such as hypoxia, abnormal lipoproteins levels, atherosclerotic conditions, etc. This study aims to investigate whether the periodontal health has an impact on the blood lipid levels. Twenty-four TM patients and sex-matched controls (C) enrolled to this study. The measurements of the periodontal parameters (gingival index, plaque index, bleeding on probing, probing depth, and clinical attachment level) were recorded. Besides, in venous blood samples the lipid profile was investigated. All of the periodontal parameters were significantly higher in TM group than in C group (P<0.05). Triglyceride and cholesterol/high density lipoprotein ratio had significant positive correlations with the periodontal parameters (P<0.05). Multiple regression analyses present significant associations between gingival index, plaque index scores, and the cholesterol/high density lipoprotein ratio (P<0.001). Our study results showed that the lipid profile of TM patients might be affected by their periodontal health. Further studies are needed to determine the lipid profile involvement magnitude and the cardiovascular disease risk caused by the periodontal health of TM patients.

The use of recombinant activated factor VII in the circumcision operation in the case of a congenital factor VII deficiency.

Congenital factor VII deficiency is a rare autosomal recessive hemorrhagic disorder and surgery is normally the cause of excessive bleeding. In this report, we describe the first case with congenital factor VII deficiency admitted to our clinics for the sunnet operation (circumcision), in which recombinant activated factor VII (rFVIIa; NovoSeven) was used to manage the bleeding. The patient was an 8-year old boy with moderate factor VII deficiency (factor VII level, 4%), and rFVIIa was administered at a dose of 20 microg/kg per dose during the circumcision operation. The same dose was repeated at 2, 4, 6, 9, 12, 15, 18, 21 and 24 h post operation. The circumcision operation could therefore be safely performed in patients with congenital factor VII using rFVIIa.

Hemoglobinopathy control program in Turkey.

Hemoglobinopathies are a very important health problem in Turkey. To date many studies have been performed but there has been no national hemoglobinopathy control program (HCP). After the Turkish National Hemoglobinopathy Council (TNHC) was created all centers, foundations, and associations were combined into one organization controlled by the Ministry of Health (MOH). The MOH and the TNHC have started to register the results of the screening of 377,339 healthy subjects from 16 different cities and the recorded average frequency of the beta-thalassemia trait was 4.3%. The highest prevalence of thebeta-thalassemia trait (13.1%) was found in the Antalya region and of the HbS trait (10%) in the Cukurova region. Next, written regulations for the Fight against Hereditary Blood Disease were published especially for preventing and treating hemoglobinopathies. The MOH and the TNHC selected 33 provinces situated in the Thrace, Marmara, Aegean, Mediterranean and South Eastern regions with a high birth prevalence of severe hemoglobinopathies. The hemoglobinopathy scientific committee was set up, a guidebook was published and a national HCP was started in these high-risk provinces.

Soft tissue density variations in thalassemia major: a possible pitfall in lumbar bone mineral density measurements by dual-energy X-ray absorptiometry.

Osteoporosis is common in patients with thalassemia major. A 16-year-old patient with thalassemia major was referred for evaluation of osteoporosis. The results of dual-energy X-ray absorptiometry in a patient with thalassemia major are presented. The patient underwent measurements of the spine in both AP and lateral position. The DXA scan of the spine in AP projection showed diffusely increased density overlying the lower thoracic and upper lumbar vertebrae. The overall density of L(2)-L(4) was 0.6465 g/cm(2) (4.93 SD as compared with young adults). The DXA scan of the spine in lateral projection showed diffusely increased density in front part of the spine. The BMD of the L(3) vertebra was 0.3669 g/cm(2) (0.30 SD as compared with young adults). It is important to interpret the images visually in order to obtain true values of BMD, and preclude invalid BMD measurements.