meatTheory and practice of meat processingТеория и практика переработки мяса2414-438X2414-441XФГБНУ «Федеральный научный центр пищевых систем им. В.М. Горбатова» РАН10.21323/2414-438X-2023-8-3-191-202meat-281Research ArticleСтатьиHistological characteristics and functional properties of red and white parts of m. semitendinosus of slaughter pigshttps://orcid.org/0000-0002-4372-6448SemenovaA. A.

Anastasia A. Semenova, Doctor of Technical Sciences, Professor, Deputy Director

26, Talalikhina str., 109316, Moscow

Tel.: +7–495–676–95–11 (105)

a.semenova@fncps.ruhttps://orcid.org/0000-0001-8923-8661PchelkinaV. A.

Viktoriya A. Pchelkina, Candidate of Technical Sciences, Leading Researcher, Experimental Clinic-Laboratory of Biologically Active Substances of Animal Origin

26, Talalikhina str., 109316, Moscow

Tel.: +7–495–676–95–11 (242)

v.pchelkina@fncps.ruhttps://orcid.org/0000-0001-7625-3838NasonovaV. V.

Victoria V. Nasonova, Doctor of Technical Sciences, Head of Department of Applied Scientific and Technological Development

26, Talalikhina str., 109316, Moscow

Tel.: +7–495–676–95–11 (307)

v.nasonova@fncps.ruhttps://orcid.org/0000-0002-8102-2900LoskutovS. I.

Svyatoslav I. Loskutov, Senior Researcher, Department of Applied Scientific and Technological Development

26, Talalikhina str., 109316, Moscow

Tel.: +7–905–214–43–23

spbsl21@gmail.comhttps://orcid.org/0000-0002-0520-7022BogolyubovaN. V.

Nadezhda V. Bogolyubova, Doctor of Biological Sciences, Leading Researcher

Podolsk, Moscow Region

Tel.: +7–496–765–11–69

652202@mail.ruhttps://orcid.org/0000-0003-4242-2239NekrasovR. V.

Roman V. Nekrasov, Doctor of Agricultural Sciences, Professor of RAS, Chief Researcher

Podolsk, Moscow Region

Tel.: +7–496–765–12–77

nek_roman@mail.ruhttps://orcid.org/0000-0003-4916-747XMotovilinaA. A.

Anna A. Motovilina, Candidate of Technical Sciences, Leading Researcher, Department of Applied Scientific and Technological Development

26, Talalikhina str., 109316, Moscow

Tel.: +7–495–676–95–11 (313)

a.motovilina@fncps.ruhttps://orcid.org/0000-0002-7403-6714BogdanovaYu. I.

Yuliya I. Bogdanova, Engineer, Department of Applied Scientific and Technological Development

26, Talalikhina str., 109316, Moscow

Tel.: +7–495–676–95–11 (360)

yu.bogdanova@fncps.ruV.M. Gorbatov Federal Research Center for Food Systems; L.K. Ernst Federal Science Center for Animal HusbandryRussian FederationL.K. Ernst Federal Science Center for Animal HusbandryRussian FederationV.M. Gorbatov Federal Research Center for Food SystemsRussian FederationV.M. Gorbatov Federal Research Center for Food SystemsРоссияV.M. Gorbatov Federal Research Center for Food SystemsRussian Federation20230910202383191202Copyright © Semenova A.A., Pchelkina V.A., Nasonova V.V., Loskutov S.I., Bogolyubova N.V., Nekrasov R.V., Motovilina A.A., Bogdanova Y.I., 20232023Semenova A.A., Pchelkina V.A., Nasonova V.V., Loskutov S.I., Bogolyubova N.V., Nekrasov R.V., Motovilina A.A., Bogdanova Y.I.Semenova A.A., Pchelkina V.A., Nasonova V.V., Loskutov S.I., Bogolyubova N.V., Nekrasov R.V., Motovilina A.A., Bogdanova Y.I.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.meatjournal.ru/jour/article/view/281

A unique muscle of pigs (Sus scrofa domesticus) is m. semitendinosus, which contains the “red” (dark) part located mainly in the depth of the leg cut and the “white” (light) part located in the close proximity to the subcutaneous fat layer. Differences in the characteristics of its “red” and “white” parts can exert a significant effect on quality and economic indicators of meat products. The aim of this research was to study histological features of the microstructure and technological properties of muscle tissue from different parts of m. semitendinosus, obtained from slaughter pigs of Russian production. M. semitendinosus was excised from chilled porcine carcasses (N=20) 24 hours after slaughter in the process of deboning. Histological examination showed that the dark part of the muscle was characterized by a higher package density of fibers, higher number of capillaries and higher sarcomere length. On the contrary, the light part was characterized by a higher diameter of muscle fibers. Analysis of muscle fiber types showed that the proportion of type I, intermediate and type IIb fibers was higher by 9.3, 5.2 and 4.1%, respectively, in the dark part. Significant differences between the dark and light parts of m. semitendinosus were revealed in terms of the number and size of giant fibers: the light part was characterized by a larger number (by more than 5 times) of giant fibers with the fibers of a larger size (almost by 11%). The samples of minced meat from the dark and light parts showed significant (р<0.05) differences in the mean values of lightness, redness and yellowness (L*, a* and b*) by 6.00, 4.68 and 3.01 units, respectively, in raw samples, and by 6.53, 2.99 and 1.81, respectively, after curing with the nitrite mixture and cooking (р<0.05). The dark part of m. semitendinosus had higher pH values (р<0.05) both for raw and cooked samples. The consistency of the samples from the light part was less elastic, looser and more crumbly than that in the samples produced from the dark part of m. semitendinosus, which was confirmed by the structural-mechanical investigations. Therefore, this study showed significant differences between the dark and light parts of m. semitendinosus by microstructural and functional-technological characteristics. Significant variability by muscle fiber diameter, which was observed in the light part of this muscle, apparently should be taken into account in breeding work and quality assessment of pork from slaughter animals.

meat qualitymeat defectsPSEhypercontractionm. semitendinosustype of muscle tissueshistologyThis research was implemented with the financial support of the Russian Science Foundation, project No. 19-16-00068-П.ReferencesSchubert-Schoppmeyer, A., Fiedler, I., Nurnberg, G., Jonas, L., Ender, K., Maak, S. et al. (2008). Simulation of giant fibre development in biopsy samples from pig longissimus muscle. Meat Science, 80(4), 1297–1303. https://doi.org/10.1016/j.meatsci.2008.06.008Schubert-Schoppmeyer, A., Fiedler, I., Nurnberg, G., Jonas, L., Ender, K., Maak, S. et al. (2008). Simulation of giant fibre development in biopsy samples from pig longissimus muscle. Meat Science, 80(4), 1297–1303. https://doi.org/10.1016/j.meatsci.2008.06.008England, E.M., Matarneh, S.K., Oliver, E.M., Apaoblaza, A., Scheffler, T.L., Shi, H. et al. (2016). Excess glycogen does not resolve high ultimate pH of oxidative muscle. Meat Science, 114, 95–102. https://doi.org/10.1016/j.meatsci.2015.10.010England, E.M., Matarneh, S.K., Oliver, E.M., Apaoblaza, A., Scheffler, T.L., Shi, H. et al. (2016). Excess glycogen does not resolve high ultimate pH of oxidative muscle. Meat Science, 114, 95–102. https://doi.org/10.1016/j.meatsci.2015.10.010Listrat, A., Lebret, B., Louveau, I., Astruc, T., Bonnet, M., Lefaucheur, L. et al. (2016). How muscle structure and composition influence meat and flesh quality. The Scientific World Journal, 2016, Article 3182746. https://doi.org/10.1155/2016/3182746Listrat, A., Lebret, B., Louveau, I., Astruc, T., Bonnet, M., Lefaucheur, L. et al. (2016). How muscle structure and composition influence meat and flesh quality. The Scientific World Journal, 2016, Article 3182746. https://doi.org/10.1155/2016/3182746Beecher, G.R., Kastenschmidt, L.L., Cassens, R.G., Hoekstra, W.G., Briskey, E.J. (1968). A Comparison of the light and dark portions of a striated muscle. Journal of Food Science, 33, 84–88. https://doi.org/10.1111/j.1365-2621.1968.tb00889.xBeecher, G.R., Kastenschmidt, L.L., Cassens, R.G., Hoekstra, W.G., Briskey, E.J. (1968). A Comparison of the light and dark portions of a striated muscle. Journal of Food Science, 33, 84–88. https://doi.org/10.1111/j.1365-2621.1968.tb00889.xStufft, K., Elgin, J., Patterson, B., Matarneh, S.K., Preisser, R., Shi, H. et al. (2017). Muscle characteristics only partially explain color variations in fresh hams. Meat Science, 128, 88–96. https://doi.org/10.1016/j.meatsci.2016.12.012Stufft, K., Elgin, J., Patterson, B., Matarneh, S.K., Preisser, R., Shi, H. et al. (2017). Muscle characteristics only partially explain color variations in fresh hams. Meat Science, 128, 88–96. https://doi.org/10.1016/j.meatsci.2016.12.012Eliášová, M., Kameník, J., Saláková, A., Pavlík, Z., Pospiech, M., Tremlová, B. (2017). The effect of PSE and non-PSE adductor and semimembranosus pig muscles on the occurrence of destructured zones in cooked hams. Journal of Food Quality, 2017(10), Article 6305051. https://doi.org/10.1155/2017/6305051Eliášová, M., Kameník, J., Saláková, A., Pavlík, Z., Pospiech, M., Tremlová, B. (2017). The effect of PSE and non-PSE adductor and semimembranosus pig muscles on the occurrence of destructured zones in cooked hams. Journal of Food Quality, 2017(10), Article 6305051. https://doi.org/10.1155/2017/6305051Safina, G.F., Chernov, V.V., Suslina, E.N., Pavlova, S.V., Kozlova, N.A., Novikov, A.A. et al. (2020). Yearbook on breeding in pig husbandry in enterprises of the Russian Federation (2019). Moscow: VNIIPlem, 2020.Safina, G.F., Chernov, V.V., Suslina, E.N., Pavlova, S.V., Kozlova, N.A., Novikov, A.A. et al. (2020). Yearbook on breeding in pig husbandry in enterprises of the Russian Federation (2019). Moscow: VNIIPlem, 2020.Semenova, A. A., Pchelkina, V. A., Nasonova, V. V., Kuznetsova, T. G., Sinichkina, A. I., Loskutov, S. I. et al. (2022). Post-slaughter state of muscle tissue of pigs depending on the duration of pre-slaughter fasting. Theory and Practice of Meat Processing, 7(3), 156–163. https://doi.org/10.21323/2414-438X-2022-7-3-156-163 (In Russian)Semenova, A. A., Pchelkina, V. A., Nasonova, V. V., Kuznetsova, T. G., Sinichkina, A. I., Loskutov, S. I. et al. (2022). Post-slaughter state of muscle tissue of pigs depending on the duration of pre-slaughter fasting. Theory and Practice of Meat Processing, 7(3), 156–163. https://doi.org/10.21323/2414-438X-2022-7-3-156-163 (In Russian)Semenova, A. A., Kuznetsova, T. G., Pchelkina, V. A., Nasonova, V. V., Loskutov, S. I., Bogolyubova, N.V. et al. (2023). Effect of adaptogens on muscle tissue microstructure of hybrid pigs (Sus scrofa domesticus L.) during intensive fattening. Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 58(2), 355–372. https://doi.org/10.15389/agrobiology.2023.2.355engSemenova, A. A., Kuznetsova, T. G., Pchelkina, V. A., Nasonova, V. V., Loskutov, S. I., Bogolyubova, N.V. et al. (2023). Effect of adaptogens on muscle tissue microstructure of hybrid pigs (Sus scrofa domesticus L.) during intensive fattening. Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 58(2), 355–372. https://doi.org/10.15389/agrobiology.2023.2.355engLefaucheur, L., Lebret, B. (2020). The rearing system modulates biochemical and histological differences in loin and ham muscles between Basque and Large White pigs. Animal, 14(9), 1976–1986. https://doi.org/10.1017/S175173112000066XLefaucheur, L., Lebret, B. (2020). The rearing system modulates biochemical and histological differences in loin and ham muscles between Basque and Large White pigs. Animal, 14(9), 1976–1986. https://doi.org/10.1017/S175173112000066XKim, G. -D., Yang, H. -S., Jeong, J. -Y. (2018). Intramuscular variations of proteome and muscle fiber type distribution in semimembranosus and semitendinosus muscles associated with pork quality. Food Chemistry, 244, 143–152. https://doi.org/10.1016/j.foodchem.2017.10.046Kim, G. -D., Yang, H. -S., Jeong, J. -Y. (2018). Intramuscular variations of proteome and muscle fiber type distribution in semimembranosus and semitendinosus muscles associated with pork quality. Food Chemistry, 244, 143–152. https://doi.org/10.1016/j.foodchem.2017.10.046Idriceanu, L., Mironeasa, S., Gheorghe, A., Lefter, N.A., Iuga, M., Grigore, D.M. et al. (2020). Effects of the extruded linseed and walnut meal on some quality characteristics of longissimus dorsi and semitendinosus muscle of pigs. Scientific Papers. Series D. Animal Science, LXIII(1), 128–134.Idriceanu, L., Mironeasa, S., Gheorghe, A., Lefter, N.A., Iuga, M., Grigore, D.M. et al. (2020). Effects of the extruded linseed and walnut meal on some quality characteristics of longissimus dorsi and semitendinosus muscle of pigs. Scientific Papers. Series D. Animal Science, LXIII(1), 128–134.Chen, F. -F., Wang, Y. -Q., Tang, G.-R., Liu, S. -G., Cai, R., Gao, Y. et al. (2019). Differences between porcine longissimus thoracis and semitendinosus intramuscular fat content and the regulation of their preadipocytes during adipogenic differentiation. Meat Science, 147, 116–126. https://doi.org/10.1016/j.meatsci.2018.09.002Chen, F. -F., Wang, Y. -Q., Tang, G.-R., Liu, S. -G., Cai, R., Gao, Y. et al. (2019). Differences between porcine longissimus thoracis and semitendinosus intramuscular fat content and the regulation of their preadipocytes during adipogenic differentiation. Meat Science, 147, 116–126. https://doi.org/10.1016/j.meatsci.2018.09.002Idriceanu, L., Mironeasa, S., Lefter, N.A., Gheorghe, A., Iuga, M., Grigore, D.M. et al. (2020). Effects of the dietary millet (Panicum Miliaceum) on some quality characteristics of liver, Longissimus dorsi and Semitendinosus muscle on pigs. Lucrări Științifice USAMV — Iași Seria Zootehnie, 73(25), 271–276.Idriceanu, L., Mironeasa, S., Lefter, N.A., Gheorghe, A., Iuga, M., Grigore, D.M. et al. (2020). Effects of the dietary millet (Panicum Miliaceum) on some quality characteristics of liver, Longissimus dorsi and Semitendinosus muscle on pigs. Lucrări Științifice USAMV — Iași Seria Zootehnie, 73(25), 271–276.Aescht, E., Büchl-Zimmermann, S., Burmester, A., Dänhardt-Pfeiffer, S., Desel, C., Hamers, C. et al. (2010). Romeis — Mikroskopische technik. Spektrum Akademischer Verlag Heidelberg, 2010. https://doi.org/10.1007/978-3-8274-2254-5 (In German)Aescht, E., Büchl-Zimmermann, S., Burmester, A., Dänhardt-Pfeiffer, S., Desel, C., Hamers, C. et al. (2010). Romeis — Mikroskopische technik. Spektrum Akademischer Verlag Heidelberg, 2010. https://doi.org/10.1007/978-3-8274-2254-5 (In German)Ertbjerg, P., Puolanne, E. (2017). Muscle structure, sarcomere length and influences on meat quality: A review. Meat Science, 132, 139–152. https://doi.org/10.1016/j.meatsci.2017.04.261Ertbjerg, P., Puolanne, E. (2017). Muscle structure, sarcomere length and influences on meat quality: A review. Meat Science, 132, 139–152. https://doi.org/10.1016/j.meatsci.2017.04.261Kasprzyk, A., Bogucka, J. (2020). Meat quality of Pulawska breed pigs and image of longissimus lumborum muscle microstructure compared to commercial DanBred and Naima hybrids. Archives Animal Breeding, 63(2), 293–301. https://doi.org/10.5194/aab-63-293-2020Kasprzyk, A., Bogucka, J. (2020). Meat quality of Pulawska breed pigs and image of longissimus lumborum muscle microstructure compared to commercial DanBred and Naima hybrids. Archives Animal Breeding, 63(2), 293–301. https://doi.org/10.5194/aab-63-293-2020Lefaucheur, L. (2010). A second look into fibre typing. Relation to meat quality. Meat Science, 84(2), 257–270. https://doi.org/10.1016/j.meatsci.2009.05.004Lefaucheur, L. (2010). A second look into fibre typing. Relation to meat quality. Meat Science, 84(2), 257–270. https://doi.org/10.1016/j.meatsci.2009.05.004Shen, L. Y., Luo, J., Lei, H. G., Jiang, Y. Z., Bai, L., Li, M. Z. et al. (2015). Effects of muscle fiber type on glycolytic potential and meat quality traits in different Tibetan pig muscles and their association with glycolysis-related gene expression. Genetics and molecular research, 14(4), 14366–14378. https://doi.org/10.4238/2015.November.13.22Shen, L. Y., Luo, J., Lei, H. G., Jiang, Y. Z., Bai, L., Li, M. Z. et al. (2015). Effects of muscle fiber type on glycolytic potential and meat quality traits in different Tibetan pig muscles and their association with glycolysis-related gene expression. Genetics and molecular research, 14(4), 14366–14378. https://doi.org/10.4238/2015.November.13.22Solomon, M.B., Van Laack, R.L.J.M., Eastridge, J.S. (1998). Biophysical basis of pale, soft, exudative (PSE) pork and poultry muscle: a review. Journal of Muscle Foods, 9(1), 1–11. https://doi.org/10.1111/j.1745-4573.1998.tb00639.xSolomon, M.B., Van Laack, R.L.J.M., Eastridge, J.S. (1998). Biophysical basis of pale, soft, exudative (PSE) pork and poultry muscle: a review. Journal of Muscle Foods, 9(1), 1–11. https://doi.org/10.1111/j.1745-4573.1998.tb00639.xFiedler, I., Nürnberg, K., Hardge, T., Nürnberg, T., Ender, K. (2003). Phenotypic variations of muscle fibre and intramuscular fat traits in Longissimus muscle of F2 population Duroc × Berlin Miniature Pig and relationships to meat quality. Meat Science, 63(1), 131–139. https://doi.org/10.1016/s0309-1740(02)00075-xFiedler, I., Nürnberg, K., Hardge, T., Nürnberg, T., Ender, K. (2003). Phenotypic variations of muscle fibre and intramuscular fat traits in Longissimus muscle of F2 population Duroc × Berlin Miniature Pig and relationships to meat quality. Meat Science, 63(1), 131–139. https://doi.org/10.1016/s0309-1740(02)00075-xSobczak, M., Lachowicz, K., Żochowska-Kujawska, J. (2010). The influence of giant fibres on utility for production of massaged products of porcine muscle longissimus dorsi. Meat Science, 84, 638–644. https://doi.org/10.1016/j.meatsci.2009.10.024Sobczak, M., Lachowicz, K., Żochowska-Kujawska, J. (2010). The influence of giant fibres on utility for production of massaged products of porcine muscle longissimus dorsi. Meat Science, 84, 638–644. https://doi.org/10.1016/j.meatsci.2009.10.024LeMaster, M.N., Warner, R.D., Chauhan, S.S., D’Souza, D.N., Dunshea, F.R. (2023). Meta-regression analysis of relationships between fibre type and meat quality in beef and pork — focus on pork. Foods, 12(11), Article 2215. https://doi.org/10.3390/foods12112215LeMaster, M.N., Warner, R.D., Chauhan, S.S., D’Souza, D.N., Dunshea, F.R. (2023). Meta-regression analysis of relationships between fibre type and meat quality in beef and pork — focus on pork. Foods, 12(11), Article 2215. https://doi.org/10.3390/foods12112215Liu, Q., Long, Y., Zhang, Y.F., Zhang, Z.Y., Yang, B., Chen, C.Y. et al (2021). Phenotypic and genetic correlations of pork myoglobin content with meat colour and other traits in an eight breed-crossed heterogeneous population. Animal, 15(1), Article 100364. https://doi.org/10.1016/j.animal.2021.100364Liu, Q., Long, Y., Zhang, Y.F., Zhang, Z.Y., Yang, B., Chen, C.Y. et al (2021). Phenotypic and genetic correlations of pork myoglobin content with meat colour and other traits in an eight breed-crossed heterogeneous population. Animal, 15(1), Article 100364. https://doi.org/10.1016/j.animal.2021.100364Chmiel, M., Słowiński, M., Dasiewicz, K. (2011). Lightness of the color measured by computer image analysis as a factor for assessing the quality of pork meat. Meat Science, 88(3), 566–570, https://doi.org/10.1016/j.meatsci.2011.02.014Chmiel, M., Słowiński, M., Dasiewicz, K. (2011). Lightness of the color measured by computer image analysis as a factor for assessing the quality of pork meat. Meat Science, 88(3), 566–570, https://doi.org/10.1016/j.meatsci.2011.02.014Kim, T.W., Kim, C.W., Kwon, S.G., Hwang, J.H., Park, D.H., Kang, D.G. et al. (2016). pH as analytical indicator for managing pork meat quality. Sains Malaysiana, 45(7), 1097–1103Kim, T.W., Kim, C.W., Kwon, S.G., Hwang, J.H., Park, D.H., Kang, D.G. et al. (2016). pH as analytical indicator for managing pork meat quality. Sains Malaysiana, 45(7), 1097–1103Gould, G.W. (1996). Methods for preservation and extension of shelf life. International Journal of Food Microbiology, 33(1), 51–64. https://doi.org/10.1016/0168-1605(96)01133-6Gould, G.W. (1996). Methods for preservation and extension of shelf life. International Journal of Food Microbiology, 33(1), 51–64. https://doi.org/10.1016/0168-1605(96)01133-6Cauble, R.N., Ball, J.J., Zorn, V.E., Reyes, T.M., Wagoner, M.P, Coursen, M.M. et al. (2021). Characteristics of pork muscles cooked to varying end-point temperatures. Foods, 10(12), Article 2963. https://doi.org/10.3390/foods10122963Cauble, R.N., Ball, J.J., Zorn, V.E., Reyes, T.M., Wagoner, M.P, Coursen, M.M. et al. (2021). Characteristics of pork muscles cooked to varying end-point temperatures. Foods, 10(12), Article 2963. https://doi.org/10.3390/foods10122963Cruzen, S.M., Baumgard, L.H., Gabler, N.K., Pearce, S. C., Lonergan, S. M. (2017). Temporal proteomic response to acute heat stress in the porcine muscle sarcoplasm. Journal of Animal Science, 95(9), 3961–3971. https://doi.org/10.2527/jas.2017.1375Cruzen, S.M., Baumgard, L.H., Gabler, N.K., Pearce, S. C., Lonergan, S. M. (2017). Temporal proteomic response to acute heat stress in the porcine muscle sarcoplasm. Journal of Animal Science, 95(9), 3961–3971. https://doi.org/10.2527/jas.2017.1375Kalbe, C., Priepke, A., Nürnberg, G., Dannenberger, D. (2019). Effects of long-term microalgae supplementation on muscle microstructure, meat quality and fatty acid composition in growing pigs. Journal of Animal Physiology Animal Nutrition, 103(2), 574–582. https://doi.org/10.1111/jpn.13037Kalbe, C., Priepke, A., Nürnberg, G., Dannenberger, D. (2019). Effects of long-term microalgae supplementation on muscle microstructure, meat quality and fatty acid composition in growing pigs. Journal of Animal Physiology Animal Nutrition, 103(2), 574–582. https://doi.org/10.1111/jpn.13037Yang, C., He, J., Yu, B., Chen, D., Mao, XB., Yu, J. et al. (2019). Long-term ingestion of low amylose/amylopectin ratio diet affects aspects of meat quality by changing muscle fibre characteristics in growing-finishing pigs. Journal of Animal Physiology Animal Nutrition, 103(2), 644–652. https://doi.org/10.1111/jpn.12967Yang, C., He, J., Yu, B., Chen, D., Mao, XB., Yu, J. et al. (2019). Long-term ingestion of low amylose/amylopectin ratio diet affects aspects of meat quality by changing muscle fibre characteristics in growing-finishing pigs. Journal of Animal Physiology Animal Nutrition, 103(2), 644–652. https://doi.org/10.1111/jpn.12967

The authors declare that there are no conflicts of interest present.