Single article

Kozlov V., Sapozhnikov S., Fufaeva A., Aleksandrova V., Petrova Yu., Mizheev M.

Bacteria as a primary source of palatine amyloid

Keywords: amyloid, thioflavin, fluorescence, palatine tonsil, chronic tonsillitis, paratonsillar abscess

The aim of the study was a pathohistological study of palatine tonsil scraping in staining with thioflavin to detect the presence of amyloid in tonsil’s surface structures as a method of minimally invasive diagnosis of amyloidosis. The material was scrapings from palatine tonsils of two young men, taken immediately before a planned (28 y.o.) and urgent (23 y.o., tonsillar abscess) tonsillectomy. Scrapings from the right and left tonsils of both patients were applied to slide glasses, stained with a 0.1% solution of thioflavin S by top irrigation and covered with coverslips. In an hour after taking the scrapings, they were examined under a luminescent microscope Lumam, fluorescence intensity of luminous objects was measured with the help of the FMEL 1A luminometer in millivolts. In both cases in tonsillar scrapings fluorescent with dark green or yellow-brown caseous masses (fluorescence intensity of 84±17 mV) were detected as well as: 1) single fluorescent epithelial cells (10±3); 2) non-fluorescent irregularly shaped large cells with a slightly fluorescent nucleus in the form of filamentous structures that are distinguished by a black cytoplasm against a fluorescent background; 3) dental amoebae with a cytoplasm and from one to four nuclei, fluorescing in green (44±49 and 86±94, respectively); 4) colonies of rod-like bacteria with bright golden-brown fluorescence (105±24); 5) cocci colonies (tetracocci and diplococci) fluorescing with green and golden yellow color (94±29 mV), and 6) single coccal forms. Based on the results of the study, conclusions are drawn: 1) mixed infection accompanying chronic tonsillitis, produces amyloid, which is contained, among others, in caseation of tonsillar crypts; 2) bacterial amyloid may be an incentive for forming amyloid masses inside a tonsil; 3) examination of scrapings and oral biopsy specimens for the purpose of identifying systemic amyloidosis is most likely to be an inadequate method of investigation; 4) detection of amyloid in a tonsillar scraping using thioflavin S can be a differentiating method of early diagnosis, testifying to the need for urgent tonsillectomy to prevent age-related systemic amyloidosis.

References

  1. Krendelev M.S. K voprosu ob etiologii tonzillita [On the issue of the etiology of tonsillitis [Electronic resource]]. Sovremennye problemy nauki i obrazovaniya, 2015, no. 4, p. 11. Available at: https://vivliophica.com/articles/medicine/314909.
  2. Rekstina V.V., Gorkovskii A.A., Bezsonov E.E., Kalebina T.S. Amiloidnye belki poverkhnosti mikroorganizmov: struktura, svoistva i znachenie dlya meditsiny [Amyloid proteins of the surface of microorganisms: structure, properties and importance for medicine]. Vestnik Rossiiskogo gosudarstvennogo meditsinskogo universiteta, 2016, no. 1, pp. 4–13.
  3. Rodina N.P., Sulatskaya A.I. Vliyanie uslovii makromolekulyarnogo kraudinga na fotofizicheskie svoistva tioflavina T – spetsificheskogo fluorestsentnogo zonda na obrazovanie amiloidnykh fibril Influence of macromolecular crowding conditions on the photophysical properties of thioflavin of a T-specific fluorescent probe on the formation of amyloid fibrils]. Sovremennye tendentsii razvitiya nauki i tekhnologii, 2015, no. 6-3, pp. 81–84.
  4. Selimzyanova L.R., Vishneva E.A., Promyslova E.A. Tonzillity u detei: voprosy patogeneza i vozmozhnosti fitoterapii [Tonsillitis in children: pathogenesis and the possibility of phytotherapy]. Pediatricheskaya farmakologiya, 2014, vol. 11, no. 4, pp. 129–133.
  5. Sulatskaya A.I., Kuznetsova I.M., Turoverov K.K. Ispol’zovanie fluorestsentnogo krasitelya tioflavina t dlya izucheniya struktury amiloidnykh fibrill [Use of fluorescent dye thioflavin t to study the structure of amyloid fibrils]. Vestnik Sankt-Peterburgskogo universiteta. Fizika i khimiya, 2011, no. 4, pp. 152–160.
  6. Khasanov S.A., Mukhrimova Sh.Z. Osobennosti mikroflory nebnykh mindalin s khronicheskim tonzillitom u detei doshkol’nogo vozrasta [Features of microflora of palatine tonsils with chronic tonsillitis in preschool children]. Molodoi uchenyi, 2016, no. 25(129), pp. 163–167.
  7. Chapman M.R., Robinson L.S., Pinkner J.S., Roth R., Heuser J., Hammar M., Normark S., Hultgren S.J. Role of Escherichia coli curli operons in directing amyloid fiber formation. Science (New York), 2002, vol. 295, pp. 851–855.
  8. Claessen D., Rink R., de Jong W., Siebring J., de Vreugd P., Boersa F.G.H., Dijkhuizen L., Wosten H.A.B. A novel class of secreted hydrophobic proteins is involved in aerial hyphae formation in Streptomyces coelicolor by forming amyloid-like fibrils. Genes & development., 2003, no. 17, pp. 1714–1726.
  9. Luheshi L., Dobson C. Bridging the gap: From protein misfolding to protein misfolding diseases. FEBS Letters., 2009, no. 583(16), pp. 2581-2586. DOI: 10.1016/j.febslet.2009.06.030.
  10. MaskevichA., Stsiapura V.I., Kuzmitsky V.A., Kuznetsova I.M., Povarova O.I., Uversky V.N., Turoverov K.K. Spectral properties of thioflavin T in solvents with different dielectric properties and in a fibril-incorporated form. J. Proteome Res.. 2007, vol. 6, no. 4. pp. 1392–1401. DOI: 10.1021/ pr0605567.
  11. Nakazato M., Matsukura S. New Type of Amyloidosis. Islet Amyloid Polypeptide(IAPP/Amylin) in Non-Insulin-Dependent Diabetes Mellitus. Internal Medicine, 1993, vol. 32, no. 12, 928–929.
  12. Sipe J.D., Benson M.D., Buxbaum J.N., Ikeda S., Merlini G., Saraiva M.J., Westermark P. Amyloid fibril protein nomenclature: 2010 recommendations from the nomenclature committee of the International Society of Amyloidosis. Amyloid., 2010, vol. 17, no. 3–4, pp. 101–104. DOI: 3109/1350 6129.2010.526812.
  13. Soscia, Kirby J., Washicosky K., Tucker S., Ingelsson M., Hyma, B., Burton M., Goldstein L., Duong S., Tanzi R., Moir R. The Alzheimer’s Disease-Associated Amyloid β‑Protein Is an Antimicrobial Peptide. PLoS ONE., 2015, no. 3, pp. e9505. DOI: 10.1371/journal.pone.0009505.
  14. Tsyrkunov V., Rybak N., Vasil’ev A., Rybak R. Microbiological and morphological aspects of chronic tonsillitis. Infekcionnye bolezni., 2016, vol. 14, no. 1, pp. 42–47. DOI: 10.20953/1729-9225-2016-1-42-47

About authors

Kozlov Vadim A.
Doctor of Biological Sciences, Candidate of Medical Sciences, Professor of the Department of Medical Biology with a course in Microbiology and Virology, Chuvash State University, Russia, Cheboksary (pooh12@yandex.ru; ORCID: https://orcid.org/0000-0001-7488-1240)
Mizheev Mikhail Borisovich
highest category doctor, otorhinolaryngological unit head, Emergency Hospital, Russia, Cheboksary (mmbmbi@yandex.ru; )
Sapozhnikov Sergey P.
Doctor of Medical Sciences, Head of the Department of Medical Biology with a course in Microbiology and Virology, Chuvash State University, Russia, Cheboksary (adaptogon@mail.ru; ORCID: https://orcid.org/0000-0003-0967-7192)
Fufaeva Alena I.
Physician, Department of Otorhinolaryngology and Maxillofacial Surgery, Republican Pediatric Clinical Hospital, Russia, Cheboksary (priffetik@bk.ru; ORCID: https://orcid.org/0000-0003-4771-7562)
Petrova Julia Victorovna
3th year student, Chuvash State University, Russia, Cheboksary (yulya-cbx@mail.ru; )
Alexandrova Vera Yurievna
3th year student, Chuvash State University, Russia, Cheboksary (verochka789@mail.ru; )

Article link

Kozlov V., Sapozhnikov S., Fufaeva A., Aleksandrova V., Petrova Yu., Mizheev M. Bacteria as a primary source of palatine amyloid [Electronic resource] // Acta medica Eurasica. – 2018. – №3. P. 24-33. – URL: https://acta-medica-eurasica.ru/en/single/2018/3/4/.