Molecular detection of circulating thyroid specific transcripts (TSHR/Tg-mRNAs) in thyroid cancer patients: Their diagnostic significance

Thyroid cancer is the most prevalent endocrine malignancy. The preoperative diagnosis of differentiated thyroid cancer (DTC) that relies solely on fine-needle aspiration (FNAC) biopsy, sometimes possesses conflicting results. New molecular markers for thyroid cancer have been investigated with most of them based on the detection in thyroid nodules or tumor tissue specimens. Recently, it was possible to detect thyroid cancer cells in the circulation by measuring the mRNA of thyroid specific genes. Among these, thyroglobulin and more recently thyroid stimulating hormone receptor mRNAs, TSHR/Tg-mRNAs in peripheral blood might serve as cancer-specific markers. These have become promising new circulating markers for thyroid cancer. The purpose of this study is to assess TSHR/Tg-mRNAs as diagnostic molecular markers for thyroid cancer and if they can be used preoperatively in synergy with FNAC. This study was performed on 60 subjects; 20 healthy volunteers and 40 patients; including 16 patients with benign thyroid diseases, 24 patients with thyroid cancer; 18 patients with newly diagnosed (DTC) and 6 patients with recurrent thyroid cancer. Diagnosis of cancer was based on FNAC and histopathology of surgical specimens. All subjects had TSHR/Tg-mRNAs in peripheral blood measured by reverse transcriptase (RT)-PCR. Based on cytology/pathology; 18 patients had newly diagnosed DTC and 11 had benign thyroid disease. Preoperative FNAC was performed on 29 of 40 patients; FNAC was diagnostic in 11/18 of malignant lesions (61.1%), in 8/11 of benign lesions (72.7%), while 10/29 (34.5%) were indeterminate. TSHR/Tg-mRNAs correctly diagnosed DTC in 20/24 and 19/24 (sensitivity 83.3% and 79.1%) and benign disease in 14/16 and 13/16 (specificity 87.5% and 81.3%), respectively. With indeterminate FNA, TSHR/Tg-mRNAs correctly diagnosed DTC (follicular type) in 5/7 and benign disease in 2/3 (combined sensitivity 71.4%; specificity 66.7%). There was high concordance between RT-PCR results for TSHR-mRNA and Tg-mRNA. Of the controls 19/20 (95%) and 16/20 (80%) were negative for both TSHR- and Tg-mRNAs. With the use of a carefully selected primer pair and qualitative RT-PCR; our results indicate that TSHR/Tg-mRNAs in peripheral blood are both equally sensitive and specific markers for detection of thyroid cancer cells. Combining TSHR/Tg-mRNAs and FNAC and ultrasound enhances the preoperative detection of cancer in patients with thyroid nodules, reducing unnecessary surgeries and correctly classified most follicular cancers and could have spared surgery in patients with benign disease.Keywords: Differentiated thyroid Cancer; TSHR/Tg-mRNAs; Fine-needle aspiration cytology; Thyroid nodules; Indeterminate lesions; Molecular marke

Pilot study for assessment of prevalence of intrafamilial hepatitis C transmission in relation to salivary viral load among infected patients with and without chronic renal failure

HCV-RNA in saliva of HCV patients provides a biological basis for its potential transmission. HCV viremia is particularly high in HCV patients on hemodialysis. This study aimed to evaluate the prevalence of HCV in saliva of HCV patients with and without renal failure, and the possible role of intrafamilial transmission of the virus. Twenty HCV patients were enrolled in this study. They were divided into two groups: 10 HCV infected patients without renal failure (Group I) and 10 with renal failure (Group II).  Detection of HCV-RNA by quantitative RT-PCR in serum and saliva of both groups was done. Thirty-eight family members of both groups were included for the detection of serum HCV antibody. The percentage of the saliva-positive patients for HCV was significantly higher in the renal failure group (70%) than the other group (40%) (p<0.05). There was insignificant statistical difference between the two groups as regards infectivity to their family contacts. Also there was insignificant correlation between the level of viremia and the intra familial transmission with a mean + SD (9,33,250 +24,501) in negative relatives and a mean + SD(79,912+26,879) in positive relatives (p> 0.05). But a significant correlation was revealed between the level of viremia and saliva positivity, with a mean + SD(12,95,666 + 1792) in saliva-positive patients and a mean +SD (3,74,465 + 2150) in saliva-negative patients (p< 0.05). There was a highly significant difference between infectivity of HCV saliva-positive patients and saliva-negative patients to their family contacts (p< 0.001). Conclusion: Increased percentage of HCV detection in saliva of HCV patients with renal failure on HD may cause spreading of HCV in HD units among RF patients. Also there was increased percentage of interfamilial infectivity among the saliva-positive patients to their relatives and this suggests that saliva might have an infective role.Keywords: Salivary; HCV; HCV PCR; Intrafamilial transmissio

Salivary PCR detection of Helicobacter pylori DNA in Egyptian patients with dyspepsia

Several methods are available for detecting Helicobacter pylori infection: (1) invasive methods based on gastric biopsies, (2) non invasive methods like Urea Breath Test (UBT), serology and stool antigen tests. Importance of salivary PCR in detection of H. pylori is still questionable. To evaluate the role of salivary PCR technique in detecting H. pylori gastric affection in Egyptian patients with dyspepsia and in differentiating between functional dyspepsia and acid-ulcer syndrome. This study included 60 patients with dyspepsia classified into three groups: (Group 1) patients with gastric H. pylori and ulcers or erosions (n= 20), (Group 2) patients with gastric H. pylori and no ulcers or erosions and had functional dyspepsia (n= 20), (Group 3) patients without H. pylori and had functional dyspepsia (n= 20). All underwent upper gastrointestinal endoscopy with biopsies, rapid urease test and salivary samples for H. pylori PCR. Significant difference between the three groups regarding salivary PCR values. No significant difference between Group 1 and Group 2 but both had significant difference with Group 3, significant difference between gastric H. pylori positive patients (n= 40) and negative ones (n= 20). Salivary PCR test had sensitivity of 85%, specificity of 70% in diagnosing H. pylori. PCR value of 534000 Iu/ml had best sensitivity (75%) and specificity (100%) for diagnosing H. pylori, highly significant positive correlation between H. pylori gastric affection and salivary PCR values. No significant difference between patients with acid ulcer syndrome (n=20) and those with functional dyspepsia (n= 40) as regard salivary PCR mean values. Salivary PCR test showed sensitivity of 100%, specificity of 50% in differentiating between patients with acid ulcer syndrome and those with functional dyspepsia. PCR value of 440000 Iu/ml had best sensitivity (100%) and specificity (55%) in differentiating acid ulcer syndrome from functional dyspepsia with non significant. H. pylori salivary PCR may be of value in diagnosing H. pylori gastric affection and is strongly correlated with it but it is of limited value in differentiating between acid ulcer syndrome and functional dyspepsia.Keywords: Salivary PCR; Helicobacter pylori; Functional dyspepsia; Acid ulcer syndrom

Conversion of barley SNPs into PCR-based markers using dCAPS method

Molecular genetic research relies heavily on the ability to detect polymorphisms in DNA. Single nucleotide polymorphisms (SNPs) are the most frequent form of DNA variation in the genome. In combination with a PCR assay, the corresponding SNP can be analyzed as a derived cleaved amplified polymorphic sequence (dCAPS) marker. The dCAPS method exploits the well-known specificity of a restriction endonuclease for its recognition site and can be used to virtually detect any SNP. Here, we describe the use of the dCAPS method for detecting single-nucleotide changes by means of a barley EST, CK569932, PCR-based marker

Role of MBL2 polymorphisms in sepsis and survival: A pilot study and in silico analysis

Sepsis is a serious infection-induced syndrome with serious ramifications, especially in intensive care units. Global concern motivated the investigation of the role of related genes’ polymorphism in predicting the liability to infection, sepsis, septic shock and survival. Among these genes is the gene encoding mannose-binding lectin (MBL), with its remarkable importance in the immune system. However, the previous studies showed conflicting results and ambiguity that urged us to engage with this issue in the Egyptian population. Prediction of functional and structural impacts of single nucleotide polymorphisms (SNPs) was done using in silico methods. A prospective observational study was conducted in intensive care units; one hundred and thirty patients were followed up. Genotyping was performed using real-time polymerase chain reaction (RT-PCR) technology. MBL SNPs showed a remarkable high frequency in our population, as well. No significant association was found between MBL2 genotypes and any of our analyses (sepsis, septic shock and survival). Only septic shock and age were independently associated with time of survival by Cox regression analysis. Our study may confirm the redundancy of MBL and the absence of significant impact on sepsis liability and mortality in adult patients

Ethnic Inequalities in Mortality: The Case of Arab-Americans

BACKGROUND: Although nearly 112 million residents of the United States belong to a non-white ethnic group, the literature about differences in health indicators across ethnic groups is limited almost exclusively to Hispanics. Features of the social experience of many ethnic groups including immigration, discrimination, and acculturation may plausibly influence mortality risk. We explored life expectancy and age-adjusted mortality risk of Arab-Americans (AAs), relative to non-Arab and non-Hispanic Whites in Michigan, the state with the largest per capita population of AAs in the US. METHODOLOGY/PRINCIPAL FINDINGS: Data were collected about all deaths to AAs and non-Arab and non-Hispanic Whites in Michigan between 1990 and 2007, and year 2000 census data were collected for population denominators. We calculated life expectancy, age-adjusted all-cause, cause-specific, and age-specific mortality rates stratified by ethnicity and gender among AAs and non-Arab and non-Hispanic Whites. Among AAs, life expectancies among men and women were 2.0 and 1.4 years lower than among non-Arab and non-Hispanic White men and women, respectively. AA men had higher mortality than non-Arab and non-Hispanic White men due to infectious diseases, chronic diseases, and homicide. AA women had higher mortality than non-Arab and non-Hispanic White women due to chronic diseases. CONCLUSIONS/SIGNIFICANCE: Despite better education and higher income, AAs have higher age-adjusted mortality risk than non-Arab and non-Hispanic Whites, particularly due to chronic diseases. Features specific to AA culture may explain some of these findings

Suicide among Arab-Americans

BACKGROUND: Arab-American (AA) populations in the US are exposed to discrimination and acculturative stress-two factors that have been associated with higher suicide risk. However, prior work suggests that socially oriented norms and behaviors, which characterize recent immigrant ethnic groups, may be protective against suicide risk. Here we explored suicide rates and their determinants among AAs in Michigan, the state with the largest proportion of AAs in the US. METHODOLOGY/PRINCIPAL FINDINGS: ICD-9/10 underlying cause of death codes were used to identify suicide deaths from among all deaths in Michigan between 1990 and 2007. Data from the 2000 U.S. Census were collected for population denominators. Age-adjusted suicide rates among AAs and non-ethnic whites were calculated by gender using the direct method of standardization. We also stratified by residence inside or outside of Wayne County (WC), the county with the largest AA population in the state. Suicide rates were 25.10 per 100,000 per year among men and 6.40 per 100,000 per year among women in Michigan from 1990 to 2007. AA men had a 51% lower suicide rate and AA women had a 33% lower rate than non-ethnic white men and women, respectively. The suicide rate among AA men in WC was 29% lower than in all other counties, while the rate among AA women in WC was 20% lower than in all other counties. Among non-ethnic whites, the suicide rate in WC was higher compared to all other counties among both men (12%) and women (16%). CONCLUSIONS/SIGNIFICANCE: Suicide rates were higher among non-ethnic white men and women compared to AA men and women in both contexts. Arab ethnicity may protect against suicide in both sexes, but more so among men. Additionally, ethnic density may protect against suicide among Arab-Americans

Role of the Endogenous Antioxidant System in the Protection of Schistosoma mansoni Primary Sporocysts against Exogenous Oxidative Stress

Antioxidants produced by the parasite Schistosoma mansoni are believed to be involved in the maintenance of cellular redox balance, thus contributing to larval survival in their intermediate snail host, Biomphalaria glabrata. Here, we focused on specific antioxidant enzymes, including glutathione-S-transferases 26 and 28 (GST26 and 28), glutathione peroxidase (GPx), peroxiredoxin 1 and 2 (Prx1 and 2) and Cu/Zn superoxide dismutase (SOD), known to be involved in cellular redox reactions, in an attempt to evaluate their endogenous antioxidant function in the early-developing primary sporocyst stage of S. mansoni. Previously we demonstrated a specific and consistent RNA interference (RNAi)-mediated knockdown of GST26 and 28, Prx1 and 2, and GPx transcripts, and an unexpected elevation of SOD transcripts in sporocysts treated with gene-specific double-stranded (ds)RNA. In the present followup study, in vitro transforming sporocysts were exposed to dsRNAs for GST26 and 28, combined Prx1/2, GPx, SOD or green-fluorescent protein (GFP, control) for 7 days in culture, followed by assessment of the effects of specific dsRNA treatments on protein levels using semi-quantitative Western blot analysis (GST26, Prx1/2 only), and larval susceptibility to exogenous oxidative stress in in vitro killing assays. Significant decreases (80% and 50%) in immunoreactive GST26 and Prx1/2, respectively, were observed in sporocysts treated with specific dsRNA, compared to control larvae treated with GFP dsRNA. Sporocysts cultured with dsRNAs for GST26, GST28, Prx1/2 and GPx, but not SOD dsRNA, were significantly increased in their susceptibility to H2O2 oxidative stress (60–80% mortalities at 48 hr) compared to GFP dsRNA controls (∼18% mortality). H2O2-mediated killing was abrogated by bovine catalase, further supporting a protective role for endogenous sporocyst antioxidants. Finally, in vitro killing of S. mansoni sporocysts by hemocytes of susceptible NMRI B. glabrata snails was increased in larvae treated with Prx1/2, GST26 and GST28 dsRNA, compared to those treated with GFP or SOD dsRNAs. Results of these experiments strongly support the hypothesis that endogenous expression and regulation of larval antioxidant enzymes serve a direct role in protection against external oxidative stress, including immune-mediated cytotoxic reactions. Moreover, these findings illustrate the efficacy of a RNAi-type approach in investigating gene function in larval schistosomes

Perspectives of the Apiaceae Hepatoprotective Effects - A Review

The liver has the crucial role in the regulation of various physiological processes and in the excretion of endogenous waste metabolites and xenobiotics. Liver structure impairment can be caused by various factors including microorganisms, autoimmune diseases, chemicals, alcohol and drugs. The plant kingdom is full of liver protective chemicals such as phenols, coumarins, lignans, essential oils, monoterpenes, carotenoids, glycosides, flavonoids, organic acids, lipids, alkaloids and xanthenes. Apiaceae plants are usually used as a vegetable or as a spice, but their other functional properties are also very important. This review highlights the significance of caraway, dill, cumin, aniseed, fennel, coriander, celery, lovage, angelica, parsley and carrot, which are popular vegetables and spices, but possess hepatoprotective potential. These plants can be used for medicinal applications to patients who suffer from liver damage

Insecticide resistance in the sand fly, Phlebotomus papatasi from Khartoum State, Sudan

<p>Abstract</p> <p>Background</p> <p><it>Phlebotomus papatasi </it>the vector of cutaneous leishmaniasis (CL) is the most widely spread sand fly in Sudan. No data has previously been collected on insecticide susceptibility and/or resistance of this vector, and a first study to establish a baseline data is reported here.</p> <p>Methods</p> <p>Sand flies were collected from Surogia village, (Khartoum State), Rahad Game Reserve (eastern Sudan) and White Nile area (Central Sudan) using light traps. Sand flies were reared in the Tropical Medicine Research Institute laboratory. The insecticide susceptibility status of first progeny (F1) of <it>P. papatasi </it>of each population was tested using WHO insecticide kits. Also, <it>P. papatasi </it>specimens from Surogia village and Rahad Game Reserve were assayed for activities of enzyme systems involved in insecticide resistance (acetylcholinesterase (AChE), non-specific carboxylesterases (EST), glutathione-S-transferases (GSTs) and cytochrome p450 monooxygenases (Cyt p450).</p> <p>Results</p> <p>Populations of <it>P. papatasi </it>from White Nile and Rahad Game Reserve were sensitive to dichlorodiphenyltrichloroethane (DDT), permethrin, malathion, and propoxur. However, the <it>P. papatasi </it>population from Surogia village was sensitive to DDT and permethrin but highly resistant to malathion and propoxur. Furthermore, <it>P. papatasi </it>of Surogia village had significantly higher insecticide detoxification enzyme activity than of those of Rahad Game Reserve. The sand fly population in Surogia displayed high AChE activity and only three specimens had elevated levels for EST and GST.</p> <p>Conclusions</p> <p>The study provided evidence for malathion and propoxur resistance in the sand fly population of Surogia village, which probably resulted from anti-malarial control activities carried out in the area during the past 50 years.</p