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1. Comparative measurement of collagen bundle orientation by Fourier analysis and semiquantitative evaluation: reliability and agreement in Masson's trichrome, Picrosirius red and confocal microscopy techniques

Author

P. Molina Aguilar, Amparo Ruiz-Sauri, Víctor Marcos-Garcés, M. Harvat, and A. Ferrández Izquierdo

Subjects
0301 basic medicine, Polarized light microscopy, Histology, Materials science, 030102 biochemistry & molecular biology, Coefficient of variation, Analytical chemistry, Magnification, Pathology and Forensic Medicine, Staining, Masson's trichrome stain, 03 medical and health sciences, symbols.namesake, 030104 developmental biology, Trichrome, Fourier analysis, Microscopy, symbols, Biomedical engineering
Abstract

Summary Measurement of collagen bundle orientation in histopathological samples is a widely used and useful technique in many research and clinical scenarios. Fourier analysis is the preferred method for performing this measurement, but the most appropriate staining and microscopy technique remains unclear. Some authors advocate the use of Haematoxylin-Eosin (H&E) and confocal microscopy, but there are no studies comparing this technique with other classical collagen stainings. In our study, 46 human skin samples were collected, processed for histological analysis and stained with Masson's trichrome, Picrosirius red and H&E. Five microphotographs of the reticular dermis were taken with a 200× magnification with light microscopy, polarized microscopy and confocal microscopy, respectively. Two independent observers measured collagen bundle orientation with semiautomated Fourier analysis with the Image-Pro Plus 7.0 software and three independent observers performed a semiquantitative evaluation of the same parameter. The average orientation for each case was calculated with the values of the five pictures. We analyzed the interrater reliability, the consistency between Fourier analysis and average semiquantitative evaluation and the consistency between measurements in Masson's trichrome, Picrosirius red and H&E-confocal. Statistical analysis for reliability and agreement was performed with the SPSS 22.0 software and consisted of intraclass correlation coefficient (ICC), Bland-Altman plots and limits of agreement and coefficient of variation. Interrater reliability was almost perfect (ICC > 0.8) with all three histological and microscopy techniques and always superior in Fourier analysis than in average semiquantitative evaluation. Measurements were consistent between Fourier analysis by one observer and average semiquantitative evaluation by three observers, with an almost perfect agreement with Masson's trichrome and Picrosirius red techniques (ICC > 0.8) and a strong agreement with H&E-confocal (0.7 0.8), better with Fourier analysis than with semiquantitative evaluation (single and average). These results in nonpathological skin samples were also confirmed in a preliminary analysis in eight scleroderma skin samples. Our results show that Masson's trichrome and Picrosirius red are consistent with H&E-confocal for measuring collagen bundle orientation in histological samples and could thus be used indistinctly for this purpose. Fourier analysis is superior to average semiquantitative evaluation and should keep being used as the preferred method.

Published
2017
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2. Poster session: Aortic stenosis

Author

R. Piccolo, J. Clarke, C. A. Brambila, B. Igual Munoz, K. Hristova, M. S. Carvalho, M. Tesic, O. Azevedo, J. A. Del Prado, A. Mcculloch, O. Kaitozis, B. Popovic, S. Stankovic, H. Chamsi-Pasha, R. Abdelfatah, V. Parisi, K. Pushparajah, E. Zemtsovsky, B. Kilickiran Avci, A. Manouras, K. Takenaka, F. Parthenakis, P. Vardas, A. Goudev, M. Orii, A. Kutarski, R. De Rosa, M. Castillo Orive, A. Sahlen, H. Ahn, S. Nedjati-Gilani, G. J. King, H. Bellsham-Revell, D. Lahidheb, M. Anastasiou-Nana, F. Pereira Machado, S. Yurdakul, N. Olsen, S. Pica, A. Ebihara, T. Nakajima, P. Molina Aguilar, R. Hornsten, M. Elnoamany, M. Cramer, G. Tamborini, G. Pagano, H. Kim, S. Soderberg, A. M. Gonzalez, N. Zlatareva, E. Marangio, F. Yang, G. Cho, I. Paunovic, C. Jons, T. Tanimoto, H. Triantafyllidi, D. Gopalan, O. Ozcan, M. Norman, G. Grazioli, F. Castillo, E. Kort, R. Bruno, J. Kostic, M. Daimon, D. Kang, C. Badiu, C. Magnino, C. Bucca, I. Joao, F. Buendia Sanchez, A. Tomaszewski, M. Alasnig, J. Kisslo, T. Kawata, S. Fernandez Casares, A. Livingston, J. Silva Cardoso, S. Korkmaz, J. Rodriguez Garcia, M. Tomaszewski, Y. Motoyoshi, A. Kaneva, E. Kinova, J. Lekakis, N. Bruun, M. Elneklawy, K. Uno, K. Nour, J. M. Ferrer, T. Wada, T. Katova, E. Ermis, F. Gaita, S. Rafla, F. Macedo, S. Woo, S. Perry, M. Lonnebakken, K. Thapa, M. Banovic, C. Selton-Suty, V. Pereira, A. Lourenco, G. Dreyfus, W. Serra, M. Hedstrom, A. Hagendorff, H. Nishino, T. Filali, M. Muratori, F. De Stefano, J. Marin, B. Jedaida, I. Rangel, J. Haertel, S. Tzortzis, A. Kalogerakis, G. Galasso, P. Hoffman, L. Chen, Y. Juilliere, V. Kostova, J. Navarro Manchon, C. J. Lopez-Guarch, J L Moya Mur, J. D. J. Baguda, C. Moretti, C. Manisty, N. Hajlaoui, H. Mahfoudhi, E. Martins, F. Bourlon, Y. Choi, C. Papadopoulos, A. Santos, I. V. Vassiliadis, A. Pereira, D. Domingo Valero, P. Iacotucci, C. Fernandez-Golfin, P. Li, I. Xanthopoulou, G. Pontone, R. Tan, D. D. Valero, D. Cramariuc, D. Lovric, F. Maffessanti, V. Pehar Pejcinovic, Y. Xu, M. Gurzun, L. Mitrofanova, P. Sousa, M. Miglioranza, A. Goncalves, I. Nedeljkovic, S. Stanic, C Di Mario, Y. Shiono, Y. Bian, E. Tossavainen, N. Risum, L. Sargento, K. Hirata, K. Said, H. Park, A. M. Argudo, T. Kubo, S. Barker, A. Chetta, R. Palma Reis, E. Malev, C. Yao, I. Papadakis, R. Medeiros, J. Tong, M. Previtali, T. Yamaguchi, S.-H. Shin, M. Sitges, C. Calinescu, J. Rueda Soriano, K. Steine, R. Ichikawa, K. Farouk, S. Pedri, J. Ripsweden, S. Carillo, G. Gelbrich, P. Rees, F. Costantino, S. Hutchings, A. Bel Minguez, A. Gaspar, M. Petrovic, M. Li Kam Wa, E. Mavronasiou, R. Winter, I. Quelhas, J. Johnson, A. Gopal, H. Jurin, R. Rordorf, M. Al-Mallah, A. Kydd, M. Ezat, A. M. Duncan, A. Kyriacou, Y. Kim, D. Mihalcea, J. Lessa, L. Mont, T. Fritz Hansen, J. Separovic Hanzevacki, D. Mesa, R. Mincu, G. Pavlidis, A.D.J. Ten Harkel, L. Gabrielli, F. Civaia, B. Vujisic-Tesic, M. Lourenco, C. Cefalu, C. Alexandrescu, L. Stefani, D. Gerede, M. Bartesaghi, C. Calin, F. Alamanni, A. Giesecke, P. Fazendas, C. Sousa, C. Ginghina, J. Magne, S. Lemoine, M. Gonzalez, C. Gohlke-Baerwolf, K. H. Hirata, S. Fawzi, H. Kisacik, B. Popescu, L. Visconti, W. Brzozowski, M. Driessen, V. Schiano Lomoriello, S. Yamada, I. Machado, F. Silveira, A. Nordin, E. Velazquez, J. Simpson, D. Vasilev, R. Rimbas, R. Murphy, C. Szymanski, T. Imanishi, M. Martirosyan, E. Najjar, J. Chambers, I. Jovanovic, A. Nagorni, E. Gunyeli, M. Omelchenko, P. De Araujo Goncalves, E. Avenatti, R. Marinov, A. Rieck, C. Tribouilloy, I. Sitges, P. Navas Tejedor, N. Lousada, W. Fehri, B. Pezo Nikolic, T. Leiner, C. Lazaro Rivera, H. Pereira, M. Loeffler, R. Hural, D. Caldeira, D. Francis, M. Di Natale, P. Salgado Filho, F. Gao, C. Alm, G. Tarsia, A. Aleixo, D. Vinereanu, C. Cotrim, M. Lotfi, B. Mc Loughlin, H. Morita, S. K. Saha, A. Djordjevic-Dikic, D. Voilliot, R. Camporotondo, J. Shin, P. Pavlov, M. A. Cattabiani, G. Sekita, A. Djordjevic Dikic, K. Ishibashi, C. Pare, J. Kwan, S. Miyazaki, V. Di Tante, E. Svenungsson, V. Giga, Y. Ino, M. Rover, J. Niewiadomska, M. Florescu, I. Skjoerten, C. Wilson, P. Davlouros, M. Hazekamp, N. Moat, A. Correia, C. Tekedis, I. Ikonomidis, B. Dilekci, L. Magda, T. Le, D. Sohn, S. Hamdy, M. Cinteza, R. Enache, A. Milan, R. Dahmani, A. Lopez Granados, J. Zamorano Gomez, E. Zorio Grima, S. Ghulam Ali, B. Demirkan, A. Shehata, M. Vono, M. Chiarlo, Miguel Mota Carmo, D. Trifunovic, B. Bijnens, Y. Yatomi, J J Jimenez Nacher, B. Rogge, R. Nagai, D. Dutka, X. Shen, I. Mordi, M. Henein, F. Celeste, G. Nadais, H. El Atroush, T. Yamano, D. Andreini, B. Beleslin, H. Suzuki, L. Yan, S. Ghio, C. C. De Sousa, S. Stoebe, S. Petrovic-Nagorni, D. Leosco, T. Komori, S. El-Tobgi, S. Mihaila, A. Madureira, T. Leiria, G. Kim, H. Haouala, B. Stuart, G. Touati, K. Oleszczak, M. Ostojic, J. Song, D. Presutti, A. Fournier, H. Daida, M. Perez Guillen, I. Kuipers, H. Hwang, B. Belesiln, K. Park, Y. Guray, D. Pfeiffer, C. Reverberi, A. Lech, A. Valentini, A. Cogo, F. Piscione, S. Negrea, S. Mezghani, V. Pilosoff, P. Sogaard, N. Blom, N. Tzemos, A. Mantovani, K. Okada, A. Turco, M. Peltier, B. Lopez Melgar, U. Guray, Q. Chen, S. Chamuleau, T. Stanton, F. Baeza, S. M. Rafla, J. Roquette, I. Almuntaser, E. Picano, D. Rusinaru, R. Kalil, R. Martin Asenjo, A. Kiotsekoglou, A. Chilingaryan, B. Candemir, P. Sonecki, A. Moulias, M. Rosca, H. Marques, A. Patrianakos, S. Sahin, J. Estornell Erill, O. Enescu, J. Spratt, P. Barbier, M. Maciel, I. Ivanac Vranesic, P. Lindqvist, T. Snow, J. Silva-Cardoso, N. Koutsogiannis, D. Ardissino, L. Zhong, K. Adamyan, L. Mccormick, A. Calin, P. Innelli, S. Yokoyama, C. Erol, P. Pabari, A. Tarr, M. Galderisi, S. Govind, B. Suran, I. Simova, E. Guyeli, T. Pinho, L. Bjornadal, B. Diaz Anton, J. Hilde, R. Sicari, C. Beladan, M. Ege, A. Zacharaki, L. Ghiadoni, A. A. La Huerta, S. Zdravkovic-Ciric, O. Huttin, K. Jensen-Urstad, F. Veglio, M. Elsedi, M. Nakabachi, P. Zinzius, D. Kim, H. Dores, A. Kakkavas, H. Badran, V. Sanchez Sanchez, E. Duo, J. Carrasco, A. Almeida, M. Virdee, M. Llemit, A. Anwar, L. Pratali, J. Monmeneu Menadas, S. Nevin, L. Fusini, F. Lombera Romero, E. Despotopoulos, E. Nyktari, G. Galanti, K. Kim, A. Van Der Hulst, H. Khachab, M. Dikic, I. Cruz, M. Melsom, J. Brugada, V. Mitic, M. Landolina, S. Turhan, V. Hansteen, D Rodriguez Munoz, J. S. De Lezo, N. Gori, Z. Baricevic, S.-P. Lee, M. Arnau Vives, S. Lee, P. Gripari, S. Humerfelt, F. Huang, T. Mikami, G. Soltan, T. Akasaka, S. Kaga, G. Penney, L. Toncelli, K. Boman, B. Basnyat, E. Kowalik, A. Bartolini, S. Georgiev, K. Shahgaldi, M. Pepi, M. Ruiz Ortiz, R. Sant'anna, H. Tsutsui, P. A. Fernandez, G. Tempesti, S. Aytekin, H. Iwano, Y. Nosir, C. Raineri, J. Rasmunsson, S. Lasarov, P. Lopez Lereu, V. Persic, F. Khan, J. Hisdal, M. Gommidh, A. Alhagoly, E. Gerdts, M. Milicia, G. Rengo, K. Kimura, F. Hakansson, M. Morenate, P. Mitev, M. Yacoub, M. Satendra, B. Kusmierczyk-Droszcz, E. Romo, R. Jankovic-Tomasevic, A. Roest, J. Stepanovic, J. Schwartz, Z. Ashour, L. Klitsie, J. Giner Blasco, M. Delgado, P. Omede, S. Mayordomo Gomez, I. Paraskevaidis, J. L. Zamorano, N. Goodfield, E. Dores, S. Davies, N. Patrascu, D. Alexopoulos, L. Donate Bertolin, D. Stanojevic, E. Psathakis, M. Dobric, P. Trivilou, H. Sasmaz, A. Marinkovic, O. Mirea, G. Sieswerda, M. Maruyama, A. M. Maceira Gonzalez, T. I. Imanishi, A. Santoro, G. Festa, R. Coma Samartin, and V. Atanaskovic

Subjects
medicine.medical_specialty, Stenosis, business.industry, Internal medicine, Cardiology, medicine, Radiology, Nuclear Medicine and imaging, General Medicine, Session (computer science), Cardiology and Cardiovascular Medicine, business, medicine.disease
Published
2012
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3. Age-related dermal collagen changes during development, maturation and ageing - a morphometric and comparative study

Author

V. García Bustos, Víctor Marcos-Garcés, C. Bea Serrano, P. Molina Aguilar, J. Benavent Seguí, Amparo Ruiz-Sauri, and A. Ferrández Izquierdo

Subjects
Adult, Male, medicine.medical_specialty, Histology, Adolescent, Human skin, Masson's trichrome stain, Young Adult, Internal medicine, Linear regression, medicine, Image Processing, Computer-Assisted, Humans, Child, Molecular Biology, Ecology, Evolution, Behavior and Systematics, Aged, Polynomial regression, Aged, 80 and over, Chemistry, Papillary dermis, Infant, Regression analysis, Cell Biology, Anatomy, Dermis, Original Articles, Middle Aged, Skin Aging, Endocrinology, Ageing, Child, Preschool, Regression Analysis, Female, Collagen, Reticular Dermis, Developmental Biology
Abstract

The tissue organisation of dermal collagen is gaining importance as a contributing factor both in development and ageing, as well as in skin maturation processes. In this work we aim to study different representative parameters of this structural organisation in 45 human skin samples of assorted ages, by means of image analysis. The variation of these parameters on the basis of age was assessed using several regression models (linear, quadratic and cubic). The area occupied by collagen was significantly reduced as a function of age in the papillary dermis (R(2) = 0.437, P < 0.0001), as well as the thickness of the collagen bundles (R(2) = 0.461, P < 0.0001), following statistical models of cubic and quadratic regression, respectively. The width of the papillary dermis increased in a significant manner over a linear regression model (R(2) = 0.26, P < 0.0001). In the reticular dermis, the cubic regression indicated a significant decline (R(2) = 0.392, P = 0.002) of the area filled with collagen according to the age. Both collagen thickness and bundle orientation parameters fit a quadratic regression over the age in a significant way (R(2) = 0.433 and R(2) = 0.334, respectively, both P < 0.0001). The width of the reticular dermis followed also a significant quadratic distribution according to age (R(2) = 0.193, P = 0.011). These parameters could partially explain the lifelong functional changes taking place in the skin and propose a baseline providing a useful entry point for future investigation.

Published
2014

4. Patología coronaria no arteriosclerótica como causa de muerte súbita en adultos: Casuística del Instituto de Medicina Legal de Valencia (1997-2005)

Author

F. Carratalá Calvo, P. Molina Aguilar, C. de la Presentación Blasco, and Jm. Ortiz Criado

Subjects
Enfermedad coronaria, business.industry, muerte súbita cardiaca, Medicine, business, patología forense, Pathology and Forensic Medicine
Abstract

Introducción y objetivos. En los países industrializados, la causa más frecuente de muerte súbita cardíaca (MSC) en el adulto es la isquemia miocárdica aguda por arterioesclerosis coronaria. Sin embargo, existen otras patologías coronarias, congénitas o adquiridas, que pueden producir isquemia miocárdica y muerte súbita: origen anómalo, hipoplasia, estenosis del ostium, aneurisma, disección, localización alta del ostium, puente miocárdico, vasculitis. El objetivo del presente trabajo es revisar la casuística de este tipo de patología coronaria no arterioesclerótica en las 8003 autopsias realizadas en el Instituto de Medicina Legal de Valencia (IMLV) entre los años 1997-2005. Método: Los criterios de inclusión de los casos para este estudio han sido: edad superior a 14 años, patología coronaria no arterioesclerótica como causa de muerte y circunstancias de la muerte compatibles con la definición de muerte súbita. Resultados: Se han encontrado 9 casos de muerte súbita en adultos con patología coronaria no arterioesclerótica como causa de muerte: 2 casos de puente miocárdico, 6 casos de disección coronaria y 1 caso de origen anómalo de coronaria. Conclusiones: La patología coronaria no arterioesclerótica, aunque es poco frecuente como causa de MSC, debemos conocerla y poder diagnosticarla macroscópicamente en la sala de autopsias. Siempre debe descartarse otra posible causa de muerte mediante los estudios complementarios químico-toxicológicos e histopatológicos. Es importante conseguir la máxima información clínica anterior al fallecimiento que pueda ayudar al diagnóstico en vida de este tipo de patologías, ya que en la mayoría de los casos sigue siendo un hallazgo postmortem.

Published
2005

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