In this work, we designed and fabricated two new “signal-on” electrochemical DNA sensors based on a sandwich-hybridization employing pyrrolidinyl peptide nucleic acid probes. The sensors comprised of a capture PNA probe (P1) immobilized on a screen-printed carbon electrode (SPCE) and an anthraquinone-labeled signaling probe (AQ-P2) designed to be partially complementary to the target DNA either at upstream (ASU) or downstream (ASD) positions on the DNA sequence hybridized to the P1 probe. In the presence of the DNA target, the ASD sensor showed a higher signal response when compared to the ASU sensor. The ASD sensor was then applied to simultaneously detect two high-risk human papillomavirus (HPV) DNA sequences. The target DNA was detected in the range of 0.5–100 nM, and the limits of detection (LODs) of 150 and 153 pM (3SDblank/slope) were obtained for HPV types 16 and 18, respectively. This developed sensor was successfully applied to detect a PCR-amplified sample derived from HPV type 16 (SiHa) and 18 (HeLa) positive human cancer cell lines. These findings are relevant for designing effective sensors, and the developed sensor can be readily applied to detect a wide range of DNA targets.

A new DNA sensor design for the simultaneous detection of HPV type 16 and 18 DNA

Emerging evidence has demonstrated the antitumor activity of allicin in various tumors. However, little study has been carried out on the functional role of allicin in cervical cancer. Our data showed that allicin suppressed cervical cancer cell viability in a time- and dose-dependent manner. Allicin treatment could reverse H2O2-induced reactive oxygen species accumulation. Meanwhile, levels of glutathione and superoxide dismutase were increased, but malondialdehyde was decreased after allicin incubation for 48 h. Furthermore, TUNEL staining showed that H2O2 treatment induced cell apoptosis, but allicin treatment could decrease cell apoptosis. Western blot assay showed that allicin could suppress the expression of nuclear factor erythroid 2-related factor 2 (NRF2) and heme oxygenase 1. We also showed that NRF2 prompted SiHa cell proliferation and reduced SiHa cell apoptosis. More importantly, allicin-inactivated phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) signaling could be partially reversed by overexpressing of NRF2. We also evaluated cell apoptosis in SiHa cells transfected with plasmid NRF2. Our data showed that allicin-induced cell apoptosis (43.5±3.8%) could largely be abolished by upregulation of NRF2 (12.3±2.08%). In summary, our data showed allicin was effective in suppressing the malignant phenotype of cervical cancer cells mainly by inhibiting the expression of NRF2, showing the potential clinical benefits of allicin in cervical cancer patients.

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Allicin suppresses the migration and invasion in cervical cancer cells mainly by inhibiting NRF2

Introduction Human papillomavirus has been reported as the etiological cause of most cervical cancers and other potentially malignant lesions. It also affects other areas, producing benign tumors on the skin. Plantar warts are a common problem found in clinical practice and share signs and symptoms with other dermatological conditions. Diagnosis of HPV infection remains a hot topic in research. Method : The present work systematically reviews the literature on the diagnostic techniques available in the clinical setting for any type of lesion produced by the virus and compares the techniques identified to those found in use for foot lesions. Results Results showed a variety of diagnostic methods, including molecular techniques, which exhibit more sensitivity than other methods but are less frequently applied to plantar lesions, where visual inspection is the most frequent method but can lead to errors. Conclusion The techniques identified need to be applied to plantar lesions to improve differential diagnosis in clinical practice. Expert opinion Research will continue to grow and a proper diagnostic technique for plantar lesions will be available in the near future.

Diagnostic techniques in HPV infections and the need to implement them in plantar lesions: A systematic review.

Electrochemiluminescence biosensor for HPV16 detection based on the adjusting of steric hindrance effect coupled with Exonuclease III amplification strategy.

OBJECTIVES Reffering to an increase in cervical cancer in the recent years, rapid, sensitive and economical detection of human papillomaviruses (HPVs) as causative agents of cervical cancer is important The traditional methods for the detection of HPVs in cervical cancer, such as pap smear, suffer from limitation and PCR has a potential to overcome the limitaitons The purpose of present research work was to identify the five important strains of HPV (16, 18, 31, 33 and 45) simultaneously by Multiplex PCR application METHODS Study was done on 100 cervical lesions of women DNA was extracted from specimens by a genomic DNA purification kit A 5-plex PCR was developed for the simultaneous detection of major HPV Five pair of new primers was designed for detection of HPV 16, 18, 31, 33 and 45 by Multiplex PCR RESULTS Among the 100 evaluated samples, 82 were found positive to HPVs In the meantime the highest rate of infection was for HPV 16 Also 30 of HPV positive samples had infections with two or more HPV types CONCLUSION Multiplex PCR assay used in present study can provide a rapid, sensitive and economical method for detection of viral infections and is applicable to small volumes of vaginal samples

Fateme Zandnia

Papers

Application of multiplex PCR for Rapid and sensitive detection of human papillomaviruses in cervical cancer.