Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica,Entamoeba dispar and Entamoeba moshkovskii

Background

Entamoeba moshkovskii and E. dispar are impossible to differentiate microscopically from the pathogenic species E. histolytica. Multiplex polymerase chain reaction (Multiplex PCR) is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment.

Methods

For detection and differentiation of the three-microscopy indistinguishable Entamoeba species in human, multiplex PCR assay using different DNA extraction methods was studied. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species.

Results

A 166-bp PCR product with E. histolytica DNA, a 580-bp product with E. moshkovskii DNA and a 752-bp product with E. dispar DNA were generated in a single-round and multiplex PCR reaction.

Conclusion

We recommend this PCR assay as an accurate, rapid, and effective diagnostic method for the detection and discrimination of these three Entamoeba species in both routine diagnosis of amoebiasis and epidemiological surveys.     

Linear Friction Welding of an AZ91 Magnesium Alloy and the Effect of Ca Additions on the Weld Characteristics

Solid-state welding offers distinct advantages for joining reactive materials, such as magnesium (Mg) and its alloys. This study investigates the effect of linear friction welding (LFW) on the microstructure and mechanical properties of cast AZ91 (Mg–9Al–1Zn) and AZ91–2Ca alloys, which (to the best knowledge of the authors) has not been reported in the literature. Using the same set of LFW process parameters, similar alloy joints—namely, AZ91/AZ91 and AZ91–2Ca/AZ91–2Ca—were manufactured and found to exhibit integral bonding at the interface without defects, such as porosity, inclusions, and/or cracking. Microstructural examination of the AZ91/AZ91 joint revealed dissolution of the Al-rich second phase in the weld zone, while the Mn containing phases remained and were refined. In the AZ91–2Ca/AZ91–2Ca joint, the weld zone retained Ca- and Mn-rich phases, which were also refined due to the LFW process. In both joint types, extensive recrystallization occurred during LFW, as evidenced by the refinement of the grains from ~1000 µm in the base materials to roughly 2–6 µm in the weld zone. These microstructural changes in the AZ91/AZ91 and AZ91–2Ca/AZ91–2Ca joints increased the hardness in the weld zone by 32%. The use of digital image correlation for strain mapping along the sample gage length during tensile testing revealed that the local strains were about 50% lower in the weld zone relative to the AZ91 and AZ91–2Ca base materials. This points to the higher strength of the weld zone in the AZ91/AZ91 and AZ91–2Ca/AZ91–2Ca joints due to the fine grain size, second phase refinement, and strong basal texture. Final fracture during tensile loading of both joints occurred in the base materials.     

Upregulation of Interleukin-6 in HPV-Positive Breast Cancer Patients

Background: Interleukin-6 (IL-6) is a well-known proinflammatory cytokine with tumor promoting capacity in various forms of malignancies including breast cancer (BC). Data highlighted the substantial role of HPV in the pathogenesis of BC. Compelling evidence suggests the contribution of HPV in carcinogenesis through triggering inflammatory cytokines such as IL-6. Objective: Here, we assessed the correlation between the presence of HPV infection and the status of IL-6 expression and serum level in BC. Methods: 72 tissue specimens including tumoral (Case; n=36) and their adjacent normal tissues (Control; n=36) were used. Nested-PCR and Real-Time PCR were employed to identify HPV DNA and assess the expression of IL-6, respectively. In addition, 72 sera samples from BC patients (n=36) and an age-matched healthy control group (n=36) were taken to measure the IL-6 serum level by ELISA. Results: Overall, the HPV DNA was detected in 19.4% (14/72) of samples. 33.33% (12/36) of cases and 5.5% (2/36) of the controls were found to be positive for HPV (P=0.003). The overexpression of IL-6 was observed in HPV+ samples compared to HPV- samples (P=0.05). However, the concentration of IL-6 serum level was remarkably different between patients and normal controls (P=0.0001). Intriguingly, IL-6 serum level was connected to the advanced clinical stage (III/IV), high grade (II/III), metastasis and, ER+ status of patients. Conclusions: Our finding indicated that the overexpression of the IL-6 may be connected to HPV infection in BC. Furthermore, the results reinforced the clinical significance and prognostic value of the serum IL-6 in BC patients.     

Can we rely on public data as a source of information for cancer registry in developing countries?

BACKGROUND/AIMS: Although a "hospital-based cancer registry" is important in improving patient care, a "population-based cancer registry" with emphasis on epidemiology is important in allocating health care resources and prioritizing public health programs. Because of its reliance on retrieved clinical and para-clinical documents, there is some limitation in registering all cancer incidents in this system, especially in developing countries. In this study we examined the possibility of using public data as a complementary source of information for recording cancers in a population-based cancer registry. METHODS: Along with the annual census in rural areas, a survey was performed in Golestan province in March 2004 to identify public awareness about cancer incidents in the community. Individuals were questioned about history of cancer in their close relatives during the last two years. Those who reported cancer in their relatives were also asked to name the main organ of involvement. A similar list was retrieved from the cancer registry at the Ministry of Health in Gorgan, and cases with upper GI (esophagus and gastric) cancer diagnosis from 21 March 2002 through 20 March 2004 were selected for this study. Finally, these two lists were compared for examining accuracy of the collected data. RESULTS: We included 137 cases in our study with rural residence and known addresses. Only 35 (25.5%) cases were reported by the relatives and among them only 20 (57.1%) relatives correctly reported the tumor location. Although we found a difference in accurate reporting of cancer incidents by year of diagnosis (more correct cases reported during the second versus the first year), the difference was not statistically significant between the two years. CONCLUSION: In this study, we examined the possibility of using public awareness about cancer incidents as a complementary source of information for a population-based cancer registry. We found that this approach is not ideal for reducing limitations. Therefore, we recommend a nationwide cancer registry to record all cancer-related information at the time of diagnosis. This strategy will reduce the need for performing retrospective surveys in collecting cancer-related information.     

Evaluation of fluticasone (flixonase) nasal spray versus beclomethasone (beconase) nasal spray in the treatment of allergic rhinitis

Although response to intranasal steroid therapy has been reported in patients with allergic rhinitis, efficacy of some nasal steroids is noteworthy.This study was undertaken to evaluate the efficacy of a two-week course of Fluticasone (Flixonase) nasal spray vs. Beclomethasone (beconase) nasal spray in patients with symptoms of allergic rhinitis referred to our clinic. This study reviewed sixty randomized studies with symptoms of allergic rhinitis which supported common aeroallergens with skin prick test. Patients received a total daily dose of nasal spray of Fluticasone (Flixonase) 100 mcg bid (N=30) compared with patients with allergic rhinitis who received a total daily dose of Beclomethasone (Beconase) 50mcg 2 puffs bid (N=30). Patients were visited before and after therapy, and efficacy of Flixonase and Beconase was evaluated by the change in nasal symptoms including: nasal discharge, nasal obstruction, nasal itching, and sneezing. After two weeks of treatment nasal symptoms of blockage, discharge, sneezing and itching were significantly better in the group treated with Fluticasone nasal spray (65%, 82%, 67%, 79% respectively (p<0.001) but after treatment with beconase nasal spray lower benefits in the nasal symptoms includes: 50%, 71%, 51%, 57% respectively. After two weeks of treatment no deleterious changes consequent to therapy were observed in nasal symptoms. 100 mcg bid Flixonase (Fluticasone) intranasal spray is more effective than 50 mcg 2 puffs bid Beconase (Beclomethasone) intranasal spray. Like asthma, allergic rhinitis is an inflammatory disease and should be managed with anti-inflammatory medication.