Knowledge, Attitude, Practices, and Preparedness of Dental Professionals in Prescribing Nicotine Replacement Therapy.

OBJECTIVE: To assess the knowledge, practice, attitude, and preparedness of dental professionals in prescribing nicotine replacement therapy (NRT). Methodology. A prevalidated voluntary web-based questionnaire was generated as a link through Google Drive and was sent to 117 dental professionals in North India using Whatsapp, Messenger, and Instagram social media platforms. A total of 94 responses were received and out of which 76 responses were analyzed (18 forms were excluded due to incomplete or duplicate responses). Frequency analysis was done using SPSS software version 21. RESULT: The participation rate was found to be 80.3%. More than half of the study population were familiar with the term NRT (77.6%) and its uses (67.1%), but approximately less than half of the total study subjects knew the duration (32.9%), cost (27.6%), dosage (25%), and contraindications (36.8%) of the NRT. Approximately 56.6% of the study participants showed a positive attitude towards helping patients to quit smoking through tobacco cessation counseling. Nearly one-fourth of the study population, i.e., 27.6%, were confident in explaining the negative impacts of tobacco, while 22.4% knew about the tobacco cessation protocol. Among the participants, only 27.6% reported that they practice NRT and out of which approximately less than 20% of the study participants were prescribing correct dose of NRT. CONCLUSION: Though study subjects had an ample knowledge regarding NRT use in tobacco cessation, it does not reflect their current attitude and preparedness. Thus, there is a need for continuing education to further train dental professionals for prescribing NRT.

Function and regulation of TRPP2 ion channel revealed by a gain-of-function mutant.

Mutations in polycystin-1 and transient receptor potential polycystin 2 (TRPP2) account for almost all clinically identified cases of autosomal dominant polycystic kidney disease (ADPKD), one of the most common human genetic diseases. TRPP2 functions as a cation channel in its homomeric complex and in the TRPP2/polycystin-1 receptor/ion channel complex. The activation mechanism of TRPP2 is unknown, which significantly limits the study of its function and regulation. Here, we generated a constitutively active gain-of-function (GOF) mutant of TRPP2 by applying a mutagenesis scan on the S4-S5 linker and the S5 transmembrane domain, and studied functional properties of the GOF TRPP2 channel. We found that extracellular divalent ions, including Ca(2+), inhibit the permeation of monovalent ions by directly blocking the TRPP2 channel pore. We also found that D643, a negatively charged amino acid in the pore, is crucial for channel permeability. By introducing single-point ADPKD pathogenic mutations into the GOF TRPP2, we showed that different mutations could have completely different effects on channel activity. The in vivo function of the GOF TRPP2 was investigated in zebrafish embryos. The results indicate that, compared with wild type (WT), GOF TRPP2 more efficiently rescued morphological abnormalities, including curly tail and cyst formation in the pronephric kidney, caused by down-regulation of endogenous TRPP2 expression. Thus, we established a GOF TRPP2 channel that can serve as a powerful tool for studying the function and regulation of TRPP2. The GOF channel may also have potential application for developing new therapeutic strategies for ADPKD.

Detection of CXCR2 cytokine receptor surface expression using immunofluorescence.

The interleukin-8 (IL-8, CXCL8) chemokine, also known as the neutrophil chemotactic factor, is a cytokine that plays a key role in inflammatory response, cell proliferation, migration, and survival. IL-8 expression is increased not only in inflammatory disorders, but also in many types of cancer, including prostate cancer. IL-8 acts as a ligand for the C-X-C chemokine receptor 2 (CXCR2) protein present on the cell plasma membrane. Binding of the IL-8 ligand to the CXCR2 receptor results in an intracellular signaling pathway mediated by GTP binding proteins coupled to the receptor itself. Knowledge of the CXCR2 expression levels facilitates the understanding of the role and function of IL-8. In this chapter, we describe a protocol that uses the immunofluorescence method and confocal microscopy to analyze the CXCR2 surface expression in human prostate cancer cells. However, this protocol is easily adaptable to analyze the surface expression of other cytokine receptors in different cell types.