Two-stage PCR assay for detection of human brucellosis in endemic areas

BACKGROUND: Brucellosis is a common zoonosis that can cause a severe febrile illness in humans. It constitutes a persistent health problem in many developing countries around the world. It is one of the most frequently reported diseases in Saudi Arabia and incidence is particularly high in the Central region, and around the city of Riyadh. The aim of this study was to evaluate a two-stage PCR assay for detection of human brucellosis particularly in endemic areas. METHODS: A total of 101 serum samples were collected from patients with acute febrile illness (AFI) of unknown cause from two different locations in the Western region of Saudi Arabia. The first location (Northern) is characterized by a nomadic rural population while the second (Central) is a modern urban city. All samples were subjected to DNA extraction and Brucella genus-specific PCR amplification using B4/B5 primers of the bcsp31 gene. Positive B4/B5 samples were subjected to multiplex species-specific Brucella PCR amplification. RESULTS: In the Northern location, 81.9% of the AFI samples were confirmed Brucella positive, while all the samples collected from the Central region proved to be Brucella negative. Samples positive for Brucella were subjected to multiplex species-specific Brucella amplification. B. abortus was detected in 10% and B. melitensis in 8% of the samples, while the majority (82%) of samples showed both B. abortus and B. melitensis. As expected, B. suis was not detected in any of the samples. CONCLUSIONS: This study concluded that a two-stage PCR assay could be useful as a rapid diagnostic tool to allow the consideration of brucellosis as a possible cause of AFI, particularly in non-urban locations. It also recommends the collection of epidemiological data for such patients to obtain further information that may help in rapid diagnosis

Association of serum asymmetric dimethyl-arginine and troponin I levels as a risk of myocardial infarction in thalassemia

Background: The current study evaluated level of serum asymmetric dimethylarginine (ADMA) and its association to cardiac biomarkers in thalassemia patients for early diagnosis of abnormality in myocardial infarction. Subjects and methods: This study was conducted on 80 subjects divided into four groups each with 20 subjects. Group I: Control: healthy subjects. Group II: Myocardial infarction: Patients with elevated serum troponin T. Group III: thalassemia patients. Group IV: thalassemia with myocardial infarction patients: Included 20 thalassemia patients with Myocardial infarction. Serum samples were subjected for assay of creatine kinase (CK:MB), Lactate dehydrogenase, troponin I ,ADMA, Serum MDA level was determined. Results: Data obtained showed that serum CKMB, LDH1, AST, Troponin T and ADMA levels were significant elevated in MI with or without Thalassemia compared with control groups. Serum MDA was statistically significantly elevated in MI with or without Thalassemia compared with control groups. The serum level of troponin T showed an area under curve (AUC) of 0.92 ,(sensitivity 91.0 % and specificity, 88%). Also, the ADMA supported the diagnostic profile, showing an AUC of 0.85 with (sensitivity, 92.0%; specificity, 91,9%). Conclusion: Serum ADMA is sensitive marker for incidence of MI in thalassemia patients.Keywords: CKMB, LDH1, AST, Troponin T, asymmetric dimethylarginie, Thalassemia

POSSIBLE CARDIOPROTECTIVE ACTION OF POMEGRANATE JUICE PUNICA GRANATUM AND PROPOLIS AGAINST MYOCARDIAL INFARCTION INDUCED IN RATS.

Background: The present study was conducted to evaluate the protective role of pomegranate juice alone or in combination with propolis extract against isoproterenol (ISO) induced myocardial infarction in rats. Material and Methods: Male Wister albino rats (n=60) weighing 220-280 g were divided into six groups each group contain ten rats; group I: negative control fed standard diet. Group (II-VI): Rats were injected subcutaneously with isoproterenol (150 mg/kg) for 3 days. Group III: Rats were given pomegranate juice 1ml /rat / day. Group IV: Rats were given propolis extract (50 mg/kg BW). Group V: Rats were given pomegranate juice + propolis extract. Group VI: Rats were given glyceronitrate (2.6mg/kg.bw/day). Results: The ISO-induced myocardial infarction in untreated rats has indicated by significant (p < 0.001) elevation of cardiac marker enzymes such as aspartate aminotransferase (AST), Lactate dehydrogenase (LDH), creatine kinase (CK), and cardiac troponin I (CnI ) when compared with control group. Rats treated with pomegranate juice and propolis extracts showed highly significant (

Association of serum asymmetric dimethyl-arginine and troponin I levels as a risk of myocardial infarction in thalassemia

Background: The current study evaluated level of serum asymmetric dimethylarginine (ADMA) and its association to cardiac biomarkers in thalassemia patients for early diagnosis of abnormality in myocardial infarction. Subjects and methods: This study was conducted on 80 subjects divided into four groups each with 20 subjects. Group I: Control: healthy subjects. Group II: Myocardial infarction: Patients with elevated serum troponin T. Group III: thalassemia patients. Group IV: thalassemia with myocardial infarction patients: Included 20 thalassemia patients with Myocardial infarction. Serum samples were subjected for assay of creatine kinase (CK:MB), Lactate dehydrogenase, troponin I ,ADMA, Serum MDA level was determined. Results: Data obtained showed that serum CKMB, LDH1, AST, Troponin T and ADMA levels were significant elevated in MI with or without Thalassemia compared with control groups. Serum MDA was statistically significantly elevated in MI with or without Thalassemia compared with control groups. The serum level of troponin T showed an area under curve (AUC) of 0.92 ,(sensitivity 91.0 % and specificity, 88%). Also, the ADMA supported the diagnostic profile, showing an AUC of 0.85 with (sensitivity, 92.0%; specificity, 91,9%). Conclusion: Serum ADMA is sensitive marker for incidence of MI in thalassemia patients

The sensitivity of Na+, K+ ATPase as an indicator of blood diseases

Background: Blood-related hereditary diseases are widespread in Eastern and SouthWestern regions of Saudi Arabia until recently. In this study, we used Na+, K+ATPase as an enzymatic indicator for the diagnosis of the diseases.Materials and methods: Individuals with different blood diseases (iron deficiency (n=13), anemia (n=14), thalassemia (n=16) and sickle cell anemia (n=12) were studied for Na+, K+-ATPase activity in the plasma membrane of red blood cell and compared with those of the healthy ones (n=20) of the same age and gender living in Jeddah, Saudi Arabia.Results: There was a significant elevation in the specific activity of Na+, K+ATPase in individuals with anemia compared with those of control (0.0094 + 0.001 nmol / mg protein/min versus 0.0061 0.001). On the other hand, there was a significant reduction in enzyme activity in thalassemia (0.0028 0.002 nmol / mg protein/min) and sickle cell anemia cases (0.0042 0.001 nmol / mg protein/min) compared to the control group. The cut off value for Na+, K+ATPase activity is 0.005 μmol Pi/minshowing 94% sensitivity and 93% specificity for the differentiation of blood abnormality.Conclusion: It can be recommended that the activity of Na+, K+-ATPase can be used for the diagnosis of individuals with blood diseases/disorders.Keywords: Na+, K+-ATPase, red blood cell, plasma membrane, iron deficiency anemia, thalassemia, sickle cell anemia, indicato

Analysis of SNPs of MC4R , GNB3 and FTO gene polymorphism in obese Saudi subjects

Background: The goal of this study was to analyze the association between the FTO rs17817449 (G&gt;T), G protein beta3 subunit (GNB3) C825T and Melanocortin 4 receptor (MC4R) A822G single nucleotide  olymorphism (SNP) with obesity in Saudi subjects.Methods: The subjects were divided into 2 groups according to BMI: Obese (BMI&gt; 29.9) and non- obese control (BMI&lt;24.9). Genotyping of the target genes were determined by polymerase chain reaction (PCR) followed by restriction fragment length polymorphism analysis (RFLP).Results: We demonstrated the association of the FTO genotype TT with increased weight, BMI and leptin levels in both males and females. However, there was no association of genotype TT with fasting blood glucose, triglycerides and cholesterol levels. Regarding GNB3 rs5443 polymorphism, the likelihood of obesity was linked to the TT genotype which was also associated with increased leptin levels. On the other hand, the SNP of MC4R A822G did not exhibit any significant association with obesity among studied subjects and showed only the presence of homozygous AA genotype.Conclusion: The polymorphism of FTO gene rs17817449 and GNB3 gene rs5443 (C825T) may be a genetic determinant of obesity in Saudi population whereas impact of MC4R Asn274Ser change could not be detected.Keywords: Obesity, FTO gene-polymorphism

The sensitivity of Na+, K+ ATPase as an indicator of blood diseases.

Background: Blood-related hereditary diseases are widespread in Eastern and SouthWestern regions of Saudi Arabia until recently. In this study, we used Na+, K+ATPase as an enzymatic indicator for the diagnosis of the diseases. Materials and methods: Individuals with different blood diseases (iron deficiency (n=13), anemia (n=14), thalassemia (n=16) and sickle cell anemia (n=12) were studied for Na+, K+-ATPase activity in the plasma membrane of red blood cell and compared with those of the healthy ones (n=20) of the same age and gender living in Jeddah, Saudi Arabia. Results: There was a significant elevation in the specific activity of Na+, K+ATPase in individuals with anemia compared with those of control (0.0094 + 0.001 nmol / mg protein/min versus 0.0061 \ub10.001). On the other hand, there was a significant reduction in enzyme activity in thalassemia (0.0028 \ub1 0.002 nmol / mg protein/min) and sickle cell anemia cases (0.0042 \ub1 0.001 nmol / mg protein/min) compared to the control group. The cut off value for Na+, K+ATPase activity is 0.005 \u3bcmol Pi/minshowing 94% sensitivity and 93% specificity for the differentiation of blood abnormality. Conclusion: It can be recommended that the activity of Na+, K+-ATPase can be used for the diagnosis of individuals with blood diseases/disorders

<i>Khalas</i> date flavonoids inhibited cell viability, induced apoptosis and expression of the pro-autophagy LC3-B gene in human hepatocellular carcinoma cells (HepG2)

Autophagy is a protective mechanism important in human diseases as cancer. We evaluated the impact of khalas date extract (KDE) (20-60 mg/mL) on cell viability, morphological changes, DNA fragmentation and gene expression of LC3B-II associated with autophagosome on HepG2 cell line. The GC/MS identification of KDE showed its high content of flavonoids including quercetin, myricetin, kaempferol and catechol. KDE reduced cell viability of HepG2 with IC50 (31.52 mg/mL). Cells treated with KDE showed two band of DNA fragments at (30 and 40 mg) indicating that KDE induced DNA damage and apoptosis in HepG2. The analysis RT-PCR data showed a 0.2-fold increase in the expression of LC3-B in the cells treated with KDE versus control. We concluded that, KDE flavonoids such as quercetin, myricetin kaempferol exhibited anticancer properties manifested by inhibition of HepG2 cell viability and induction of apoptosis and upregulation of the pro-autophagy LC3-B gene. </p

Identification of Deregulated Signaling Pathways in Jurkat Cells in Response to a Novel Acylspermidine Analogue-N-Erucoyl Spermidine

Natural polyamines such as putrescine, spermidine, and spermine are crucial in the cell proliferation and maintenance in all the eukaryotes. However, the requirement of polyamines in tumor cells is stepped up to maintain tumorigenicity. Many synthetic polyamine analogues have been designed recently to target the polyamine metabolism in tumors to induce apoptosis. N 4 -Erucoyl spermidine (designed as N 4 -Eru), a novel acylspermidine derivative, has been shown to exert selective inhibitory effects on both hematological and solid tumors, but its mechanisms of action are unknown. In this study, RNA sequencing was performed to investigate the anticancer mechanisms of N 4 -Eru-treated T-cell acute lymphoblastic leukemia (ALL) cell line (Jurkat cells), and gene expression was examined through different tools. We could show that many key oncogenes including NDRG1, CACNA1G, TGFBR2, NOTCH1,2,3, UHRF1, DNMT1,3, HDAC1,3, KDM3A, KDM4B, KDM4C, FOS , and SATB1 were downregulated, whereas several tumor suppressor genes such as CDKN2AIPNL, KISS1, DDIT3, TP53I13, PPARG, FOXP1 were upregulated. Data obtained through RNA-Seq further showed that N 4 -Eru inhibited the NOTCH/Wnt/JAK-STAT axis. This study also indicated that N 4 -Eru-induced apoptosis could involve several key signaling pathways in cancer. Altogether, our results suggest that N 4 -Eru is a promising drug to treat ALL