A Novel Trypsin Kunitz-Type Inhibitor from Cajanus cajan Leaves and Its Inhibitory Activity on New Cancer Serine Proteases and Its Effect on Tumor Cell Growth.

A novel trypsin inhibitor from Cajanus cajan (TIC) fresh leaves was partially purified by affinity chromatography. SDS-PAGE revealed one band with about 15 kDa with expressive trypsin inhibitor activity by zymography. TIC showed high affinity for trypsin (Ki = 1.617 µM) and was a competitive inhibitor for this serine protease. TIC activity was maintained after 24 h of treatment at 70 °C, after 1 h treatments with different pH values, and ß-mercaptoethanol increasing concentrations, and demonstrated expressive structural stability. However, the activity of TIC was affected in the presence of oxidizing agents. In order to study the effect of TIC on secreted serine proteases, as well as on the cell culture growth curve, SK-MEL-28 metastatic human melanoma cell line and CaCo-2 colon adenocarcinoma was grown in supplemented DMEM, and the extracellular fractions were submitted salting out and affinity chromatography to obtain new secreted serine proteases. TIC inhibited almost completely, 96 to 89%, the activity of these serine proteases and reduced the melanoma and colon adenocarcinoma cells growth of 48 and 77% respectively. Besides, it is the first time that a trypsin inhibitor was isolated and characterized from C. cajan leaves and cancer serine proteases were isolated and partial characterized from SK-MEL-28 and CaCo-2 cancer cell lines. Furthermore, TIC shown to be potent inhibitor of tumor protease affecting cell growth, and can be one potential drug candidate to be employed in chemotherapy of melanoma and colon adenocarcinoma.

Methodological Approaches for Vocal Folds Experiments in Laryngology: A Scoping Review.

OBJECTIVE: The aim of this study was to describe the methodological approaches adopted in experimental researches in laryngology intervention studies. METHODS: The study was performed as a scoping review using the electronic databases Medline (PubMed), Web of Science, Embase, BioMed Central and SCOPUS. These databases were manually searched from 1995 (or its inception) until the most recently published articles in June 2022. The inclusion criteria were as follows: a) studies performed with animal models in voice and/or larynx therapy, intervention and/or surgery; b) studies with participant populations composed with animals; c) studies containing original research; d) studies performed with at least one objective measurement for treatment and or intervention; e) studies reporting at least one method of larynx intervention; and f) publications written in English, Spanish or Portuguese. The exclusion criteria were as follows: a) studies considering without any objective intervention and or treatment; b) studies without animal models; and c) studies that reviewed articles or books. These criteria were set to increase inter-study comparability. RESULTS: There were found 26 studies that showed that in experimental laryngology. There were retrieved four main characteristics in the retrieved studies. It was observed five major groups of experimental models used for assessing vocal folds: dog (38.5%), rat/mouse (23,1%), pig (23.1%), rabbit (19.2%), human (11.5%), and sheep (3.8%). In addition, three characteristics were observed: sample (up to 20 subjects 88.5%), type of surgery/intervention (100%) and duration (up to 30 days 61.5%). CONCLUSION: In experimental laryngology, the state of the art is grounded on mainly in dog, rat and pig in vocal folds assessment. Up to 20 subjects, surgery/intervention and experimental trial lasting no more than 30 days are frequent methodological approaches in this research field.

Benign paroxysmal positional vertigo: diagnosis and treatment.

Benign paroxysmal positional vertigo is a common disorder in Neurotology. This vestibular syndrome is characterized by transient attacks of vertigo, caused by change in head position, and associated with paroxysmal characteristic nystagmus. The symptoms result from movement of the free floating otoconia particles in the endolymph or their attachment to the cupulae of the semicircular canal. The diagnosis is essentially clinical and should be confirmed by performing diagnostic maneuvers. Treatment is based on the identification of the affected semicircular canal and performance of liberatory maneuvers or repositioning of free floating particles of otoliths. The effectiveness varies from 70 to 100%.