Biotecnologia Aplicada, vol. 16, no. 04, 1999
BIOSCI Administrator
CC Vol. 16 No. 4 (October-December, 1999)
CC ISSN 0864-4551 (printed)
CC ISSN 1027-2852 (electronic)
CC Copyright 1999, Elfos Scientiae.
CC Founded in 1983 and published as Interferon y
CC Biotecnologia until 1990, Biotecnologia Aplicada is a
CC peer-reviewed scientific journal. It appears four times
CC per year, sponsored by the Ibero-Latin-American Society
CC of Biotechnology Applied to Health.
CC BIOTECNOLOGIA APLICADA is a vehicle of Iberian-Latin-
CC American scientists working in different fields of modern
CC biotechnology for the diffusion of their experiences and
CC results. Some of the main topics that are considered for
CC publication are the following:
CC a) Molecular genetics and recombinant DNA technology;
CC b) Therapeutic and prophylactic uses of molecules
CC obtained by biotechnological methods;
CC c) Immunotechnology and development of diagnostic
CC methods;
CC d) Chemistry, biochemistry and design of molecules with
CC biological activity;
CC e) Application of recombinant DNA techniques and its
CC products in agriculture and animal science;
CC f) Transgenesis;
CC g) Fermentation, recovery and purification of biotech-
CC nological products;
CC h) Scaling-up, plant design and economic evaluation;
CC i) Control and automation of processes.
CC All rights reserved. No part of this publication may be
CC reproduced, stored or transmitted in any form or by any
CC means without the prior permission of the Publisher. By
CC submitting a manuscript the authors agree that the
CC copyright for their article is transferred to the
CC Publisher if and when the article is accepted for
CC publication. The points of view expressed in the Journal
CC are exclusive responsibility of the authors.
CC This journal is indexed by the following publications:
CC Biosis, Current Biotechnology Abstracts, Derwent Biotech-
CC nology Abstracts, Cambridge Scientific Abstracts,
CC Chemical Abstracts, Biological Abstracts, EMBASE/Excerpta
CC Medica, Elsevier BIOBASE/Current Awareness in Biological
CC Sciences, LilaCS, Periódica, Bio-Journals (Internet).
CC Correspondence and subscriptions
CC Ave. 31 entre 158 y 190,
CC Cubanacan, Playa
CC Ciudad de La Habana,
CC Apdo. 6072, Habana 6, Cuba.
CC Telephones: (53-7) 33 1917 / 21 8466 / 21 8164
CC Fax: (53-7) 33 1917 / 21 8070 / 33 6008
CC E-mail: elfos.s...@cigb.edu.cu
CC Full papers are available online through BIOLINE System
CC (Internet) through the WWW or by e-mail. Annual subscrip-
CC tion or single document purchase are possible. Contact
CC the following for further information:
CC URL:http://www.bd.org.br/bioline/
CC E-mail address: b...@biostrat.demon.co.uk
AU Rosa A Serra, Maureen McDonnell, April Bragg
IN Department of Molecular and Cellular Physiology.
University of Cincinnati School of Medicine.
Cincinnati, OH 45267, USA. E-mail: ser...@email.uc.edu
Department of Cell Biology,
Vanderbilt University Medical Center.
Nashville, TN, USA 37232.
TI Transforming Growth Factor-beta in Development
and Disease
DE BMP
DE differentiation
DE mechanism of action
DE signaling
DE Smad
DE TGF
SO Biotecnologia Aplicada 1999;16:205-218
AB The transforming growth factor-beta (TGF-beta)
superfamily consists of multifunctional
peptides that control many aspects of cell
growth and differentiation. Because of the
multifunctional nature of the TGF-betas, an
understanding of the mechanism of TGF-beta
action should provide significant information
regarding the pathogenesis of many diseases.
This review will focus on the role and
mechanisms of TGF-beta signaling in the mammary
gland, skin, lung, and skeleton and will
discuss how alterations in TGF-beta signaling
may result in disease.
CC Review article
CC Language: English
AU Nieves Santos, Jorge Aguiar, Julio Fernandez,
Maria Vazquez, George Auburger, Suzana Gispert,
Yssel Mendoza, Julia Garcia, Luis Velazquez
IN Department of Neurology. Lenin Hospital. Holguin, Cuba.
Department of Pharmaceutics.
Center for Genetic Engineering and Biotechnology.
PO Box 6162, Havana 10600, Cuba.
Phone (53-7) 21 8008; 336008; E-mail: jag...@cigb.edu.cu
Department of Neurology. University Hospital.
Düsseldorf, Germany.
TI Molecular diagnosis of a sample of the Cuban population
with spinocerebellar ataxia type 2
DE CAG repeat expansions
DE disease
DE spinocerebellar ataxia 2
SO Biotecnologia Aplicada 1999;16:219-221
AB CAG repeat unstable expansions causing spinocerebellar
ataxia 2 (SCA2) disease were localized into a CpG island
on the exon 1 of the SCA2 gene, previously linked to the
chromosomic region 12q23-24.1 of human chromosome 12. At
molecular level, 392 patients with dominant ataxia from
Holguin, Cuba, were analyzed, showing an inverse
correlation between CAG repeat length and the age of
disease onset. The smallest CAG repeat expansion causing
neurodegeneration among the SCA2 patients was also found.
For SCA2 patients with (CAG)n stretches of 32 to 40
repeats, there is a high variability with respect to the
age of disease manifestation. These findings suggest that
for each patient other specific genetic and/or
environmental factors are very significant for the
begining and evolution of the disease for small sizes of
CAG repeat expansions.
CC Research Article
CC Language: Spanish
AU Angel Valdivia, Danay Chacon, Clara Savon, Odalys Valdes,
Grehete Gonzalez, Reynel Cancio, Julio C Sanchez,
German Roges, Blanca Garcia-Barreno, Jose A Melero,
Angel Goyenechea
IN Deparment of Virology. Institute of Tropical Medicine
"Pedro Kouri". PO Box 601, Marianao 13, Havana, Cuba.
Fax: (53-7) 24 6051, 22 0633;
E-mail: a.val...@ipk.sld.cu
Center for Genetic Engineering and Biotechnology.
PO Box 6162. Havana 6, Cuba.
National Center of Fundamental Biology, Majadahonda 28220
Madrid, Spain.
TI Confirmation of Unusual Epidemiological Behavior of the
Human Respiratory Syncytial Virus During Two Consecutive
Epidemics (Seasons 94/95 and 95/96) in Havana City, Cuba
DE HRSV
DE molecular epidemiology
DE respiratory syncytial virus
SO Biotecnologia Aplicada 1999;16:222-225
AB Twenty-seven strains of the human respiratory syncytial
virus, isolated from three consecutive outbreaks, were
sequenced at the National Center of Fundamental Biology
(Madrid, Spain). The nucleotide sequence showed two
unique characteristics related to what had been
previously reported in the literature: (i) all of them
were identical, and (ii) all showed a great homology with
the Spanish Long reference strain (with only five
changes). Upon this unusual finding and with the main
objective of excluding a probable contamination with
another Long strain from Sweden, selected regions of both
reference strains and other laboratory strains were
sequenced. The Spanish Long strain was only used as a
confirmatory control of the results obtained in
laboratories from Spain. The results obtained by the
authors discarded any possible contamination when the
percentages of homology between the sequenced regions of
the two foreign strains and the Cuban isolates were
analyzed and compared. The results also corroborated that
the Swedish Long strain was even more separated from the
Cuban isolates than the Spanish strain. This report
presents a theory to explain the unusual epidemiological
behavior of this virus in Cuba.
CC Research article
CC Language: English
AU Santiago Dueñas-Carrera, Juan Morales,
Nelson Acosta-Rivero, Lazaro J Lorenzo, Ciro Garcia,
Thelvia Ramos, Ivis Guerra, Maxlenin Peña
IN Centro de Ingenieria Genetica y Biotecnologia.
AP 6162, CP 10600, Habana, Cuba. Fax: (53-7) 214764;
E-mail: juan.m...@cigb.edu.cu
TI Variable Level Expression of Hepatitis C Virus Core
Protein in a Prokaryotic System. Analysis of the Humoral
Response against It in Rabbits
DE E. coli
DE expression
DE HCV
DE hepatitis C virus
DE immunogenicity
DE nucleocapsid core protein
DE purification
SO Biotecnologia Aplicada 1999;16:226-231
AB Experimental evidence suggests that the hepatitis C virus
(HCV) core protein has several biological properties and
is implicated as a viral factor in HCV-mediated
pathogenesis. In this study, CoE1.339, Co.176 and Co.120
variants of the HCV capsid protein were produced at
variable levels in Escherichia coli from pNCoE1, pN12 and
pSLCo120 plasmids, respectively. The chimera CoE1.339
spans the first 339 amino acids of the viral polyprotein,
fused to the 45 amino acids stabilizer peptide from P64K
protein of Neisseria meningitidis. Co.176 is a non-fused
variant encompassing amino acids 1-176 of the capsid
protein sequence. Both CoE1.339 and Co.176 were expressed
under the transcriptional control of the tryptophan
promoter and the protein was only detected by Western
blot. However, Co.120, a variant truncated at the C
terminus which contains amino acids 1-120 of the HCV core
protein fused to a leader tag of six histidines, was
successfully produced under the control of the T7
promoter in BL21 (DE3) cells. Like Co.176 and CoE1.339,
Co.120 was efficiently recognized by human anti-HCV
positive serum. Purified Co.120 antigen showed to be
immunogenic in rabbits. Synthetic peptides covering amino
acids 1-40 of the core protein showed the greatest
reactivity to the rabbit anti-Co.120 serum.
CC Research Article
CC Language: English
AU Diana Garcia del Barco, Alina Rodriguez, Elsa Rodriguez,
Caridad Tamayo, Julio R Fernandez, Maria M Vasquez,
Maria P Rodriguez, Ricardo Lleonart
IN Division de Genetica de Celulas de Mamiferos.
Centro de Ingenieria Genetica y Biotecnologia.
AP 6162, CP 10600, La Habana, Cuba. Telf: (53-7) 21 8164;
Fax: (53-7) 33 6008; E-mail: diana....@cigb.edu.cu
TI Erythropoietin gene co-amplification in
methotrexate-resistant Chinese hamster ovary dhfr- cells.
DE CHO
DE EPO
DE erythropoietin
DE gene amplification
SO Biotecnologia Aplicada 1999;16:232-235
AB Gene amplification plays an important role in cellular
regulatory mechanisms. To study the amplification and
expression of the human erythropoietin (EPO) gene,
Chinese hamster ovary (CHO) dhfr- cells co-transfected
with chimeric genes for the expression of EPO and
dihydrofolate reductase (DHFR) and selected in the
presence of increasing concentrations of methotrexate,
were used. The results evidenced a three-fold gene
amplification, accompanied by an increase in EPO
expression of up to 33 pg/cell/day. The amplicon, which
contains host genomic sequences, shows rearrangements
originated during the integration-amplification process
CC Research Article
CC Language: Spanish
AU Lisset Herrera, Orlene Guerra, Pedro L Ramos,
Rudy Peral, Ana L Echemendia, Nadia Ramirez,
Vivian Doreste, Pedro Oramas
IN Laboratory of Biochemistry. Bioplant Center.
University of Ciego de Avila. Carretera a Moron Km 9,5.
CP 69450. Ciego de Avila, Cuba.
Plant Biotechnology Division.
Center for Genetic Engineering and Biotechnology.
PO Box 6162, Havana, Cuba. Fax: (53-7) 21 8070;
E-mail: Pedro....@cigb.edu.cu
Instituto de Sanidad Vegetal.
Playa, Ciudad de La Habana, Cuba.
TI Molecular Techniques for the Detection of Tomato Yellow
Leaf Curl Geminivirus in Infected Plants and Viruliferous
Whiteflies
DE Dot blot
DE geminiviruses
DE PCR
DE TYLCV
DE virus diagnosis
SO Biotecnologia Aplicada 1999;16:237-241
AB The tomato yellow leaf curl virus (TYLCV) causes major
yield losses in tomato production in many tropical and
subtropical regions. Therefore, there is an increased
need to apply molecular methods for the detection and
characterization of the main TYLCV isolates that affect
Cuban tomato plantations. A DNA fragment containing the
gene encoding the coat protein of a Cuban TYLCV isolate
was amplified by polymerase chain reaction (PCR). DNA
hybridization with specific radiolabeled probes
corroborated the identity of the amplified product, and
allowed to analyze the integration into a full-length
TYLCV genome. The threshold of TYLCV detection by PCR in
plants and viruliferous whiteflies was within the
reported range. To detect TYLCV infections, samples
collected from tomato plantations of several regions in
Cuba were analyzed by PCR. The amplified coat protein
gene was also effectively used as a DNA probe in Dot blot
assays to detect geminivirus in plants
CC Paper on Techniques
CC Language: English
AU Guillermo Pratta, Roxana Zorzoli, Liliana A Picardi
IN Catedra de Genetica. Facultad de Ciencias Agrarias.
Universidad Nacional de Rosario. CC 14, 2123 Zavalla,
Argentina. Telf: (54-0341) 497 0080;
Fax: (54-0341) 497 0085; E-mail: gpr...@fcagr.unr.edu.ar
TI Obtainment and micropropagation of intra- and
interspecific hybrids of tomato (genus lycopersicon)
DE embryo rescue
DE in vitro plant tissue culture
DE plant breeding
DE plant genetic resources
SO Biotecnologia Aplicada 1999;16:242-245
AB In vitro plant tissue culture is a useful tool for
overcoming the problems in obtaining hybrids (i.e.,
embryo abortion, production of seeds) when wild species
are incorporated to breeding programs. The ability for
producing intra- and interspecific hybrids among
different crosses in the genus Lycopersicon, the adequacy
of in vitro culture of immature embryos to break the
unilateral incompatibility, and the micropropagation
response of the hybrids obtained, were evaluated.
Different genotypes of the species and varieties of L.
esculentum, L. esculentum var. cerasiforme, L.
pimpinellifolium, L. peruvianum, and L. hirsutum, were
used. Differences in the ability to produce intra- and
interspecific hybrids were found among the crosses. In
vitro culture of immature embryos was successful in
obtainintg interspecific hybrids when the crosses
presented unilateral incompatibility. In this case, there
also were differences in the ability to cross among the
genotypes. The capacity for in vitro regeneration of the
intra- and interspecific hybrids originated from
compatible crosses was in general superior to that of the
parental lines, but a decrease in the regeneration
capacity was observed in the hybrids originated from
incompatible crosses, with respect to the parental
genotypes.
CC Paper on Techniques
CC Language: Spanish
AU Mauro Alfonso, Peter Ifversen, Jesper Zeuthen
IN Center of Molecular Immunology. PO Box 16040,
Havana 11600, Cuba. E-mail: ma...@ict.cim.sld.cu
Department of Tumor Cell Biology. Danish Cancer Society,
DK-2100 Copenhagen, Denmark.
TI Human Anti-ganglioside IgG Antibody Response Induced
by in vitro Immunization with Liposomes Containing
GM3(NeuGc) Ganglioside
DE ganglioside
DE human antibody
DE in vitro immunization
SO Biotecnologia Aplicada 1999;16:246-248
AB A new in vitro immunization system of human peripheral
blood lymphocytes using liposomes containing
gangliosides, is described. Purified human B lymphocytes
were cultured in the CD40 system in the presence of
liposomes containing different concentrations of
GM3(NeuGc) ganglioside, ranging from 1 to 1000 ng/mL.
After an initial primary in vitro immunization period (3
4 days), antigen-stimulated B cells and autologous
activated CD4+ T lymphocytes previously activated with an
anti-CD3 monoclonal antibody were cultured in the
presence of staphylococcal enterotoxin A. Human antibody
production was measured by an anti-ganglioside ELISA
after 4 and 8 days of culture. Specific antibody
responses against GM3(NeuGc) of IgM and IgG isotypes were
induced after 4 days in culture, and a predominant
antigen-specific IgG antibody production was observed 8
days after antigenic stimulation.
CC Article on Techniques
CC Language: Spanish
AU Gerardo Guillen
IN Centro de Ingenieria Genetica y Biotecnologia.
Ave. 31 entre 158 y 190, Cubanacan, Playa. AP 6162,
CP 10600, Cuba. Telf: (53-7) 21 6221;
Fax: (53-7) 21 4764; E-mail: gerardo...@cigb.edu.cu
TI XI Congreso Internacional de Virologia
SO Biotecnologia Aplicada 1999;16:249-252
CC Report
CC Language: Spanish
AU Gerardo Guillen
IN Centro de Ingenieria Genetica y Biotecnologia.
Ave. 31 entre 158 y 190, Cubanacan, Playa. AP 6162,
CP 10600, Cuba. Telf: (53-7) 21 6221;
Fax: (53-7) 21 4764; E-mail: gerardo...@cigb.edu.cu
TI IX Congreso Internacional de Bacteriologia y Micologia
SO Biotecnologia Aplicada 1999;16:253-255
CC Report
CC Language: Spanish
AU Maria L Genta, Hugo D Genta
IN Laboratorio de Tecnologia de Alimentos,
Facultad de Bioquimica, Quimica y Farmacia.
Instituto de Ingenieria Quimica.
Facultad de Ciencias Exactas y Tecnologia.
Universidad Nacional de Tucuman.
Ave. Independencia 1800, 4000 San Miguel de Tucuman,
Argentina. Telf: (54-381) 436 4093 (Interno 222);
Fax: (54-381) 436 3004;
E-mail: nalv...@herrera.unt.edu.ar
TI La bioindustria en Japon
SO Biotecnologia Aplicada 1999;16:257-260
CC Focus
CC Language: Spanish
AU Jorge Bacallao, Josefina Lugo
IN Dpto. Computacion y Biometria.
Instituto Superior de Ciencias Medicas.
Calle 146 No. 3102, La Habana 11600, Cuba.
E-mail: baca...@comput.giron.sld.cu
Dpto. Ensayos Clinicos.
Instituto Nacional de Oncologia y Radiobiologia.
Ciudad de La Habana, Cuba.
TI Use of prior Knowledge in Randomized Clinical Trials
with a Binary Response Variable
DE beta-binomial distribution
DE binary response
DE clinical trials
DE non-Bayesian methods
SO Biotecnologia Aplicada 1999;16:261-263
AB When testing a new drug, the researcher faces the ethical
and technical problem of incorporating the existing
knowledge derived from previous trials, from a
meta-analysis or simply from his subjective judgement.
This paper proposes a non-Bayesian method based on the
beta-binomial distribution, which can be used in the
clinical trial of a drug or treatment when the response
variable is dichotomous. The method consists in the
modeling of the previous knowledge by means of the beta
distribution in the assessment of both toxicity and
efficacy, and a statistical test of efficacy, which
depends on a parameter of the beta-binomial distribution.
CC Focus
CC Language: Spanish