Identification of Entamoeba histolytica in Patients with Suspected Amebiasis in Jordan Using PCR-based Assays.

BACKGROUND AND OBJECTIVE: Identification of Entamoeba histolytica (E. histolytica) by microscopy alone can be problematic because E. dispar and E. moshkovskii are morphologically similar to E. histolytica. Therefore, this study aimed to assess the performance of microscopy in the detection of E. histolytica in stool specimens with the help of PCR-based assays and enzyme-linked immunosorbent assay (ELISA). MATERIALS AND METHODS: Between September, 2017 and September, 2018, 200 stool specimens were obtained from Jordanian patients with suspected amebiasis. All specimens were subjected to microscopic analysis. DNA was extracted from the microscopy-positive stool samples. A conventional PCR and a duplex real-time PCR were performed to detect E. histolytica and E. dispar. RESULTS: By microscopy, 35% (70/200) of specimens were tested positive for Entamoeba complex. All 70 microscopic-positive Entamoeba complex samples were negative for the presence of E. histolytica by the NOVITEC® E. histolytica ELISA assay. All 70 samples positive by microscopy were negative for the presence of E. histolytica and E. dispar by PCR-based assays. CONCLUSION: We suspect some of these microscopy-positive stool specimens might contain a potentially novel species of Entamoeba that could not be detected by ELISA or PCR-based assays specific for E. histolytica and E. dispar. Diagnosis of amebiasis remains challenging here in Jordan and hence highlighting the need for improved diagnostic method.

Cq1 Exon Polymorphisms in Caucasian and African American Systemic Lupus Erythematosus patients.

BACKGROUND: C1q protein is composed of three protein chains (A, B and C) that are the products of separate genes. Genetic deficiencies in C1Q genes are important factors influencing the risk of systemic lupus erythematosus (SLE). Therefore, this study aimed to investigate the possible association of single nucleotide polymorphisms (SNPs) in the coding region of the C1Q genes with SLE. METHODS: To search for potential SNPs in the encoding regions of C1q A, B and C chains, Cq1 exons were initially amplified and directly sequenced from leukocyte DNA from a subset of Caucasian and African American SLE patients and healthy controls. The sequences were analyzed by the Phrap and Phred software analysis system and the SNPs were identified by visual inspection. To test if any of these SNPs were linked to susceptibility to SLE, they were measured in 210 SLE patients ((59 African Americans and 151 Caucasians) and 129 matched healthy controls (55 African Americans and 74 Caucasians) by restriction fragment length polymorphism analysis. RESULTS: The sequencing phase of the study identified three synonymous SNPs: Nucleotide 276G>A in C1QA, 66C>A in C1QB and 129G>A in C1QC. Statistically, no differences were found in genotype or allele frequencies between patients and controls for the 276G>A or 66C>A SNP. However, in Caucasians, the frequencies of the 129G>A genotypes were significantly different between SLE patients and controls (P = 0.005), specifically with the GG genotype being over represented in the controls (P = 0.004). CONCLUSION: The results show that the homozygous 129GG genotype is associated with protection against SLE onset. This protection is race dependent, being observed in Caucasians but not African Americans. The mechanism of this association is currently unclear.

Phenol content, antioxidant capacity and antibacterial activity of methanolic extracts derived from four Jordanian medicinal plants.

This study was performed to assess the antioxidant and antibacterial properties of methanolic extracts derived from aerial parts of four Jordanian medicinal plants (Artemisia sieberi, Peganum harmala, Rosmarinus officinalis (Green-Flowered) and Sarcopterium spinosium). The possible relationship between these biological properties and the total phenolic concentrations of these extracts were also be determined. The antioxidant capacity and total phenolic concentrations were assessed by the ABTS method and Folin-Ciocalteu method, respectively. The amount of the extract required to scavenge 50% of ABTS (IC50) was also measured. Broth dilution and disc diffusion assays were performed to measure the antibacterial activity of these extracts against available bacterial strains. Variations were observed among the examined plants in antioxidant and antibacterial activities as well as in their phenol contents. According to ABTS assay and IC50 value, the highest free radical scavenging potential was found in Sarcopterium spinosium, followed by Rosmarinus officinalis, Peganum harmala and Artemisia sieberi, respectively. Similarly, the results of antibacterial assays showed that Sarcopterium spinosium exhibited the highest antibacterial activity against all tested bacterial strains as compared to Rosmarinus officinalis, Peganum harmala and Artemisia sieberi. Moreover, Sarcopterium spinosium contained the highest amount of phenolic compounds followed by, Rosmarinus officinalis, Artemisia sieberi and Peganum harmala, respectively. In conclusion, these plants are not only interesting sources for antimicrobial agents but also have a considerable amount of antioxidants. In addition, these findings revealed that the antioxidant capacity and antibacterial activity of these plant extracts do not necessary be attributed to their total phenolic concentrations.

Loss of P16 Protein Expression and Its Association with Epstein-Barr Virus LMP-1 Expression in Hodgkin's Lymphoma.

BACKGROUND: Expression of Epstein-Barr virus Latent Member Protein-1 (EBV LMP-1) and loss of P16 protein expression are documented in lymphoma, indicating a relationship between them, but this relationship is not clear and sometimes contradictory. Thus, this study was conducted to examine the relationship between the loss of P16 and EBV LMP-1 expression in Jordanian patients diagnosed with lymphoma. METHODS: Sections were made from archival formalin-fixed and paraffin-embedded blocks from 55 patients diagnosed with lymphoma. P16 expression and LMP-1 expression were detected by immunohistochemistry using monoclonal antibodies. RESULTS: In Hodgkin's Lymphoma (HL), the loss of P16 was higher in LMP-1 positive cases (61%) than LMP-1 negative cases (25%; P = 0.072). Conversely, in Non-Hodgkin's Lymphoma (NHL), none of LMP-1 positive samples showed loss of P16. Furthermore, among LMP-1 HL positive cases, the loss of P16 was more frequent in male (75%) than female (33%). Also, there was a significantly higher proportion of LMP-1 positive cases showing loss of P16 in HL (11:18), compared to those in NHL (0:8, P < 0.001), confirming a difference between HL and NHL, concerning the LMP-1/P16 relationship. CONCLUSION: A trend for an association between loss of P16 and LMP-1 expression was observed in HL but not NHL patients. These findings suggest that there are molecular and clinical differences in the pathogenesis and development of different subtypes of lymphoma.

Hepatoprotetive, Cardioprotective and Nephroprotective Actions of Essential Oil Extract of Artemisia sieberi in Alloxan Induced Diabetic Rats.

The aim of the current study is to evaluate the potential mechanism of antidiabetic action of the essential oil of Artemisia sieberi and its effects on some hematological and biochemical parameters in alloxan induced diabetic rats. Extraction of the essential oil from aerial parts of A. sieberi was preformed by hydrodistillation. Fifty rats were divided into five groups. Groups I and II normal rats given 1 mL/day of dimethyl sulfoxide and 80 mg/kg BW of this oil extract, respectively. Groups III, IV and V diabetic rats given 1 mL/day of dimethyl sulfoxide, oil extract (80 mg/kg BW) and metformin (14.2 mg/kg BW), respectively. Several hematological and biochemical parameters were assessed. Oral administration of the extract resulted in a significant reduction in the mean values of blood glucose, glucagon, cholesterol, triglyceride, LDL-C, ESR, urea, uric acid, creatinine accompanied by an increase in the mean values of the total protein, albumin, insulin, HDL-C, neutrophile count and PCV in diabetic rats. No significant changes in these parameters were found in the control group. The effects produced by this extract were closely similar to a standard antidiabetic drug, metformin. In conclusion, the present study indicates that the essential oil extract of A. sieberi appears to exhibit cardioprotective, nephroprotective and hepatoprotective activities in alloxan induced diabetic rats.

A polymorphism in the type one complement receptor (CR1) involves an additional cysteine within the C3b/C4b binding domain that inhibits ligand binding.

The type one complement receptor (CR1) contains a variable number of binding domains for C3b and C4b, formed through a nearly identical set of repeating units known as short consensus repeats (SCRs). Each SCR contains four cysteines that, by forming two disulfide bonds, impart a conformation critical for function. In this study, we identified a CR1 single nucleotide polymorphism (1597C>T) that results in an additional cysteine (483R>C) in SCR 8 of the N-terminal C3b/C4b binding domain, and occurring sporadically in corresponding SCRs of other repeated C3b/C4b binding domains. The normal carrier frequency for 483-C was 6.3% in 175 African Americans, and 2.4% in 153 Caucasians. In expression constructs containing one C3b/C4b binding domain, the 483-C residue reduced binding to C3b, C3bi, and C4b by over 80% (each p<0.0001), versus the wildtype construct. Full-length CR1 from 483-C carriers also exhibited reduced binding to C3b and C4b, although the effect was influenced by the total number of binding domains present. Race-matched comparisons between SLE patients (86 African Americans, 228 Caucasians) and the normal cohort showed that 483-C carrier status alone is not a risk factor for SLE or lupus nephritis. The physiological role of this polymorphism remains to be determined.

Smooth muscle cell expression of a constitutive active form of human Rac 1 accelerates cutaneous wound repair.

BACKGROUND: Hyperoxia has been shown to improve wound healing; however, the mechanism for such therapeutic effects of oxygen remains hypothetical. Rac 1 regulates a wide variety of cellular activities, including cell proliferation and migration, and also is a key regulator for the activity of the nicotinamide dinucleotide phosphate oxidase the enzyme complex responsible for the production of a large fraction of cellular superoxide. METHODS: We generated transgenic mice that express either the cDNA of a constitutively active mutant of human Rac 1 (V12 mutant or Rac CA) or the dominant negative isoform (V12 and N17 mutant or Rac DN) in the blood vessels using mouse vascular smooth muscle promoter for alpha-actin. We placed 2 wounds of 6 mm in diameter at the middorsal region of each mouse and allowed about 3 weeks for the wounds to heal. RESULTS: The size of the wounds in Rac CA transgenic mice was reduced relative to wild type mice; healing of Rac DN mice was slower than wild type and Rac CA ( P < .05). Blood vessel formation appeared faster in Rac CA mice, a finding associated with enhanced expression of some angiogenic growth factors. CONCLUSION: The current studies suggest that Rac 1 activation accelerates the wound healing process and is associated with more efficient angiogenesis at the wound site.