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Rayner Rodrı́guez

University of Miami

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María G. Guzmán

Instituto de Medicina Tropical “Pedro Kourí”

Mayling Álvarez

Instituto de Medicina Tropical “Pedro Kourí”

Aı́da Zulueta

Istituti Clinici Scientifici Maugeri

Ricardo Silva

Centro de Ingeniería Genética y Biotecnología

Laura Lazo

Centro de Ingeniería Genética y Biotecnología

Lisset Hermida

Zhejiang Yongning Pharma (China)

Delfina Rosario

Instituto de Medicina Tropical “Pedro Kourí”

Gerardo Guillén

Centro de Ingeniería Genética y Biotecnología

Iris Valdés

Centro de Ingeniería Genética y Biotecnología

Carlos López

Centro de Ingeniería Genética y Biotecnología

Cooperative Institutions

Instituto de Medicina Tropical “Pedro Kourí”

University of Miami

Centro de Ingeniería Genética y Biotecnología

Pan American Health Organization (Cuba)

Instituto de Medicina Tropical

National Center for Genetic Engineering and Biotechnology

Latin American School of Medicine

Instituto Politécnico Nacional

ORCID

Miami Transplant Institute

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Comparative studies of the action of 5-bromodeoxyuridine and 5-hydroxymethyldeoxyuridine on the growth of Escherichia coli thymidine auxotroph

Revista Latinoamericana de Microbiologia (1981)

Rayner Rodrı́guez M. Espinosa Lara B. Pena Valdivia

Auxotrophy

Thymidine

Bromodeoxyuridine

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Deterioration and Variability of Highly Purified Collagenase Blends Used in Clinical Islet Isolation

Transplantation (2007)

Toshiyuki Yamamoto Camillo Ricordi Shari Messinger Yasunaru Sakuma Atsushi Miki

Background. The quality and stability of enzyme blends used in islet cell processing are critical for successful human islet isolation. A wide variability in enzymatic activity among lots of Liberase HI has been reported. This study examines the interlot and intralot variability of Liberase HI and the over-time deterioration of enzyme quality based on the analysis of islet isolation outcomes. Methods. The data of 169 human isolations processed for clinical islet transplantation, using five different lots of Liberase HI, were retrospectively analyzed. Inter- and intralot variables in the islet isolation were assessed over a 15-month period. Results. The analysis revealed significant interlot differences in the digestion time, prepurification islet counts, percent recovery, viability, and glucose stimulation insulin index. Moreover, a significant decrease in the pre- and postpurification islet yield per pancreas weight (IEQ/g) in isolations processed by two different enzyme lots used over a 15-month period was observed, suggesting a progressive deterioration of enzyme quality. Conclusions. Our data demonstrate a significant lot-to-lot related variability in islet isolation outcomes. In addition, the over-time decline in isolation outcomes processed using a single enzyme lot was observed even when the enzyme blends were used within the expiration dating specified by the manufacturer.

Digestion

10.1097/01.tp.0000284979.48497.de

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Rat DNA polymerase beta substitutes the repairing activity of DNA polymerase I in the lethal effect of UV light.

R. Escamilla J.M Espinosa Lara J.D Quintana Hau Sarai Luna P Loyola Abitia

The aim of this work was to search if the rat DNA polymerase beta can substitute the capability of DNA polymerase I to repair damage caused by the UV light in Escherichia coli. The oriC origin of replication from p beta 5 was replaced by the rep origin from pSC101 and named p beta 6. The presence of pol beta in the new construct was verified by PCR. E. coli polA-1 (WP6) was transformed with p beta 6. A protein with size similar to DNA Pol beta (40 kDa) was shown in the cell free extracts carrying pbeta5. In WP6/p beta 6 cell free extracts a slightly smaller protein was observed instead of the 40 kDa. DNA Pol beta was revealed by western analysis, with polyclonal antibodies, in strains with p beta 5. Yet, it was not detected in the western from WP6/p beta 6. A moderate change in UV resistance was observed in strains carrying p beta 5. However, in polAl carrying p beta 6 (WP6/p beta 6), irradiated with 60-90 J/m2 of UV light, the viability was increased by more than four orders of magnitude, when compared with the polA1 (WP6) strain, reaching approximately the same UV resistance as the strains with DNA polymerase I. The results suggests that probably Pol beta is rapidly degraded in the cell free extracts from WP6/p beta 6 and, it repairs the lethal effect of the UV light in E. coli.

DNA polymerase beta

BETA (programming language)

DNA polymerase I

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