RESUMEN
Spinal muscular atrophy is a progressive motor neuron disorder caused by deletions or point mutations in the SMN1 gene. It is not known why motor neurons are particularly sensitive to a decrease in SMN protein levels and what factors besides SMN2 underlie the high clinical heterogeneity of the disease. Here we studied the methylation patterns of genes on sequential stages of motor neuron differentiation from induced pluripotent stem cells derived from the patients with SMA type I and II. The genes involved in the regulation of pluripotency, neural differentiation as well as those associated with spinal muscular atrophy development were included. The results show that the PAX6, HB9, CHAT, ARHGAP22, and SMN2 genes are differently methylated in cells derived from SMA patients compared to the cells of healthy individuals. This study clarifies the specificities of the disease pathogenesis and extends the knowledge of pathways involved in the SMA progression.
Asunto(s)
Células Madre Pluripotentes Inducidas/fisiología , Neuronas Motoras/fisiología , Atrofia Muscular Espinal/genética , Diferenciación Celular , Células Cultivadas , Metilación de ADN , Proteínas Activadoras de GTPasa/genética , Proteínas Activadoras de GTPasa/metabolismo , Regulación del Desarrollo de la Expresión Génica , Humanos , Neurogénesis , Factor de Transcripción PAX6/genética , Factor de Transcripción PAX6/metabolismo , Proteína 1 para la Supervivencia de la Neurona Motora/genética , Proteína 1 para la Supervivencia de la Neurona Motora/metabolismo , Proteína 2 para la Supervivencia de la Neurona Motora/genética , Proteína 2 para la Supervivencia de la Neurona Motora/metabolismoRESUMEN
The COVID-19 coronavirus pandemic has spread to 215 countries around the world and caused tens of millions of infections and more than a million deaths worldwide. In the midst of COVID-19 infection, it is extremely important to identify new protein and gene targets that may be highly sensitive diagnostic and prognostic markers of the severity and outcome of the disease for combating this pandemic. Identification of individual genetic predisposition allows personalizing programs of medical rehabilitation and therapy. It has now been shown that the transmissibility and severity of COVID-19 infection can be affected by gene variants in both the human body (ACE2, HLA-B*4601, FcγRIIA, MBL, TMPRSS2, TNFA, IL6, blood group A antigen, etc.) and the virus itself (ORF8 in RNA polymerase, ORF6 in RNA primase, S, N, E proteins). The presence of mutations in the proteins of the virus can change the affinity and specificity for the binding of targeted drugs to them, being the molecular basis of individual differences in the response of the human body to antiviral drugs and/or vaccines. The review summarizes the data on the variants of the genomes of the coronavirus and humans associated with an individual predisposition to an increased or decreased risk of transmission, severity, and outcome of COVID-19 infection. Targeted drugs and vaccines being developed for the therapy of COVID-19 infection are briefly reviewed.
RESUMEN
Duchenne muscular dystrophy (DMD) is a severe and rapidly progressive hereditary muscular disease with X-linked recessive inheritance, occurring mainly in males. A complete loss of dystrophin resulted from out-of-frame deletion mutations in the DMD gene leads to Duchenne muscular dystrophy. DMD induced pluripotent stem cells (iPSCs) are a suitable cell model to study muscle development and disease mechanisms underlying muscular dystrophy and to screen novel compounds with potential therapeutic effects. We generated iPSCs from a DMD patient using non-integrating episomal plasmid vectors. The obtained iPSC lines showed ESC-like morphology, expression pluripotency markers, displayed a normal karyotype and possessed trilineage differentiation potential.
Asunto(s)
Células Madre Pluripotentes Inducidas , Distrofia Muscular de Duchenne , Diferenciación Celular , Distrofina/genética , Humanos , Masculino , Distrofia Muscular de Duchenne/genéticaRESUMEN
Spinal muscular atrophy (SMA) is a genetic disease, which characterized by the degeneration of motor neurons in the spinal cord and further striated muscle atrophy. The research of the processes in diseased neurons is complicated due to the impossibility of obtaining them safely from patients. Thus, we generated SMA type III induced pluripotent stem cell lines via using non-integrated episomal plasmid vectors. The resulting cell line expresses the major pluripotency markers and can differentiate in vitro into derivatives of three germ layers. The iPSC line can be used for further studies by providing in vitro the relevant cell types.
Asunto(s)
Células Madre Pluripotentes Inducidas , Atrofia Muscular Espinal , Atrofias Musculares Espinales de la Infancia , Línea Celular , Humanos , Neuronas Motoras , Atrofia Muscular Espinal/genéticaRESUMEN
Uterine leiomyoma (UL) is the most common benign tumor in women of reproductive age. Gene therapy using suicidal genes appears to be a promising approach for UL treatment. One of key factors for success of gene therapy is the right choice of genetic construct carrier. A promising group of non-viral carriers for cell delivery of expression vectors is cationic Cys-flanked peptides which form tight complexes with DNA due to electrostatic interactions and the presence of interpeptide disulfide bonds. The paper reports a comparative study of the physico-chemical, toxic, and transfectional properties of the DNA-peptide complexes obtained by matrix polymerization or oxidative polycondensation of Cys-flanked peptides using the chain growth terminator 2-amino ethanethiol. We have demonstrated the therapeutic effect of the delivery of the pPTK-1 plasmid carrying the herpes simplex virus type 1 (HSV-1) thymidine kinase gene into PANC-1, and HEK-293T cell culture as well as into primary UL cells. It has been shown that the carriers obtained by oxidative polycondensation transform primary UL cells more efficiently than those produced by matrix polymerization. Treatment with ganciclovir resulted in the death of up to 40% of UL cells transfected with the pPTK-1 plasmid. The perspectives of use of the polyR6 carrier produced by oxidative polycondensation as a tool for the development of modular peptide carriers for the purposes of UL gene therapy were discussed.
Asunto(s)
Genes Transgénicos Suicidas , Terapia Genética , Vectores Genéticos , Leiomioma , Timidina Quinasa , Femenino , Células HEK293 , Humanos , Leiomioma/terapia , Péptidos , Simplexvirus/enzimología , Timidina Quinasa/genéticaRESUMEN
We studied the effect of peptide AEDG on telomere length and mitotic index of PHA-stimulated blood lymphocytes from young (18-22 years, N=5) and middle-aged (49-54 years, N=6) men. In the younger age group, no significant changes in the mitotic index were detected, while in the middle-aged group, a decrease in this parameter was found in one case. The relative length of telomeric regions of metaphase chromosomes was evaluated by in situ fluorescence hybridization with DNA probes specific to telomeres. After incubation with peptide AEDG, significant changes in the relative telomere length were found in 7 of 11 individuals (3 cases in the younger age group and 4 cases in the middle age group). Significant increase in telomere length after exposure to peptide AEDG was revealed in 5 cases, including two individuals of the younger age group (by 41 and 55%) and three individuals of the middle age group (by 156, 18, and 76%). In one individual of the younger age group and in one of the middle-age group, a significant decrease in telomere length (by 37 and 15%, respectively) was found. A tendency to normalization of telomere lengths was noted: this parameter increased in individuals with initially lower telomere length relative to the group mean value and decreased in individuals with initially longer telomeres compared to the mean length in the group.
Asunto(s)
Linfocitos/efectos de los fármacos , Linfocitos/metabolismo , Fitohemaglutininas/farmacología , Telómero/efectos de los fármacos , Telómero/metabolismo , Adolescente , Adulto , Humanos , Hibridación Fluorescente in Situ , Activación de Linfocitos/efectos de los fármacos , Persona de Mediana Edad , Índice Mitótico , Adulto JovenRESUMEN
Spinal muscular atrophy (SMA) is a neuromuscular disease caused by deletion or mutation in SMN1 gene. SMA human induced pluripotent stem cells (iPSCs) represent a useful and valid model for the study of the disorder, as they provide in vitro the target cells. We generated iPSCs from a SMA type I patient and SMA type II patient by using non-integrating episomal plasmid vectors. The resulting iPSCs are episomal-free, express pluripotency markers, display a normal karyotype, retain the mutation (homozygous deletion of SMN1) and are able to differentiate into the three germ layers.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Células Madre Pluripotentes Inducidas/patología , Atrofia Muscular Espinal/patología , Adulto , Línea Celular , Niño , HumanosRESUMEN
Using immunofluorescence with specific antibodies, we analyzed DNA hydroxymethylation in uncultured cells from 25 human uterine leiomyomas considering the menstrual cycle phase during surgery and the presence of MED12 gene mutations. It was found that each tumor node had specific DNA hydroxymethylation level that did not depend on the presence of mutations in MED12 gene, but depended on the phase of menstrual cycle. The degree of DNA hydroxymethylation was significantly lower in cells of leiomyomas excised during the luteal phase compared to the follicular phase (p=0.0431). Hormonal status changing at various phases of menstrual cycle is a factor affecting DNA hydroxymethylation in leiomyoma cells.
Asunto(s)
Análisis Mutacional de ADN/métodos , Hidroxilación/fisiología , Leiomioma/metabolismo , Complejo Mediador/genética , Ciclo Menstrual/genética , Neoplasias Uterinas/genética , Adulto , Femenino , Humanos , Hidroxilación/genética , Ciclo Menstrual/fisiología , Persona de Mediana Edad , Mutación/genética , Programas Informáticos , Neoplasias Uterinas/metabolismoRESUMEN
Analysis of variations in DNA structure using a low-density microarray technology for routine diagnostic in evidence-based medicine is still relevant. In this work the applicability of 3-D macroporous monolithic methacrylate-based platforms for detection of different pathogenic genomic substitutions was studied. The detection of nucleotide replacements in F5 (Leiden G/A, rs6025), MTHFR (C/T, rs1801133) and ITGB3 (T/C, rs5918), involved in coagulation, and COMT (C/G, rs4818), TPH2 (T/A, rs11178997), PON1 (T/A rs854560), AGTR2 (C/A, rs11091046) and SERPINE1 (5G/4G, rs1799889), associated with pregnancy complications, was performed. The effect of such parameters as amount and type of oligonucleotide probe, amount of PCR product on signal-to-noise ratio, as well as mismatch discrimination was analyzed. Sensitivity and specificity of mutation detections were coincided and equal to 98.6%. The analysis of SERPINE1 and MTHFR genotypes by both NGS and developed microarray was performed and compared.
Asunto(s)
Genoma Humano , Metacrilatos/química , Análisis de Secuencia por Matrices de Oligonucleótidos , Complicaciones del Embarazo/genética , Arildialquilfosfatasa/genética , Secuencia de Bases , Catecol O-Metiltransferasa/genética , Glicoles de Etileno , Femenino , Genotipo , Humanos , Integrina beta3/genética , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Mutación , Inhibidor 1 de Activador Plasminogénico/genética , Porosidad , Embarazo , Triptófano Hidroxilasa/genéticaRESUMEN
The overview represents the recent most conspicuous findings in aging studies. It includes new data on the whole genome association studies (GWAS) in big cohort of centenaries, recently found mutation protecting from Alzheimer disease, discovery of hypothalamus as a command center of human aging, very important data on the negative effect of common antioxidants in the treatment of lung cancer as well as new data concerning antiaging and anticancer effects of common drugs such as rapamycine and metformin. Substantial part of the review is devoted to the epigenetic problems of senescence and feasible impact of basic epigenetic mechanisms (methylation of DNA and histone proteins, DNA heterochromatization) in regulation of gene expression, long-term genome reprogramming during early childhood, and transgeneration transmission of epigenetic traits. The necessity of transition from molecular studies of dormant human genome (anatomy of human genome) to genome in action (dynamic genome) and thus with special emphasis to epigenetic medicine is stressed.
Asunto(s)
Envejecimiento/fisiología , Epigénesis Genética/fisiología , Fenómenos Genéticos/fisiología , Genoma Humano/fisiología , Envejecimiento/genética , Interacción Gen-Ambiente , Estudio de Asociación del Genoma Completo , Geriatría/métodos , Geriatría/tendencias , Humanos , Neoplasias/genética , Neoplasias/terapiaRESUMEN
In 2009-2010, 98 patients diagnosed with the coronary heart disease, but without the expressed metabolic violations, decompensated conditions and diseases were surveyed. Patients of 60-90 years were divided by age into two groups: younger than 75 years--47 people; 75 years and older--51; there were 41 women and 57 men; the ratio between women and men was 1:1.4; average age was 76.0 ± 1.3 years. The average age of women was 76.0 ± 1.8 years, men--76.1 ± 2,0 years. 96.0 ± 2.0% of geriatric patients with CHD complained of steno-cardiac pains in pre-cardiac area; the excess mass of a body was observed in 43.4 ± 5.0% of geriatric patients with CHD, acute myocardial infarction in the anamnesis was noted in 52.5 ± 6.9% (n = 52); hypertensive illness had 98.0 ± 1.4% (n = 97). Patients had the favorable average levels of lipoproteins of high density (not lower than 1 mmol/l). A significant number (79.2 ± 5.6%) of patients had hemodynamically significant narrowing of coronary vessels (75% and more), while the area of myocardial hypokinesia was observed in 41.2 ± 6.9% of patients, sharp violation of brain blood circulation was noted in 14.1 ± 9.8%. Wild type homozygous genotype of TCF7L2 gene was detected significantly more often (77.6 ± 4.7%) in patients of advanced and senile age with CHD, regardless of age group, exis-tence of accompanying diseases and conditions, such as previous myocardial damage, acute disorders of cerebral circulation and fatty degeneration of the liver. However allele of risk T (totally C/Tand T/T) of TCF7L2 gene came to light in 22.4 ± 9.1% of geriatric patients with CHD, that contributes to development of a metabolic syndrome in such patients and reduces term of their life.
Asunto(s)
Envejecimiento/genética , Enfermedad Coronaria/genética , Hígado Graso/genética , Evaluación Geriátrica/métodos , Polimorfismo de Nucleótido Simple , Proteína 2 Similar al Factor de Transcripción 7/genética , Anciano , Anciano de 80 o más Años , Glucemia/análisis , Comorbilidad , Enfermedad Coronaria/epidemiología , Hígado Graso/sangre , Hígado Graso/epidemiología , Femenino , Frecuencia de los Genes , Hemodinámica , Heterocigoto , Homocigoto , Humanos , Lipoproteínas HDL/sangre , Masculino , Persona de Mediana EdadRESUMEN
Genotype and allele frequencies of uncoupling proteins 2 and 3 genes (UCP2 and UCP3) and peroxisome proliferator-activated receptors genes (PPARA, PPARD and PPARG) were studied in 206 residents of the siege and in 139 individuals of more than 69 years old (control group). Studied polymorphisms included UCP2 (Ala55Val), UCP3 (C-55T), PPARA (G/C), PPARD (+294T/C), and PPARG (Pro12Ala). The G allele and the G/G genotype (PPARA) were overrepresented in the group of survivors and C/C (UCP3) genotype prevailed in the women of besieged Leningrad compared to relevant control groups of the persons of the same age who did not suffered hungry disaster. Feasible protective effects of PPARA gene allele G and C allele of UCP2 genes are briefly discussed.
Asunto(s)
Canales Iónicos/genética , Longevidad/genética , Desnutrición/genética , Proteínas Mitocondriales/genética , PPAR alfa/genética , Polimorfismo de Nucleótido Simple , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Ciudades , Interpretación Estadística de Datos , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Masculino , PPAR delta/genética , PPAR gamma/genética , Federación de Rusia , Sobrevivientes , Proteína Desacopladora 2 , Proteína Desacopladora 3 , Segunda Guerra MundialRESUMEN
We present a comparative analysis of the allelic polymorphism of the matrix metalloproteinase (MMP) gene family, including MMP3 (rs3025058), MMP7 (rs11568818), MMP9 (rs17576, rs2250889), MMP12 (rs2276109), and MMP13 (rs2252070), in patients with external genital endometriosis (EGE) and in a control group of healthy women proven to be free of disease by laparoscopic inspection. We found significant differences in the incidence of particular MMP3 and MMP9 alleles, which substantiate the role of matrix metalloproteinases in EGE pathogenesis. We used the Multifactor Dimensionality Reduction (MDR) analysis to show that 14 allelic combinations of the MMP containing MMP3 (rs3025058) x MMP7 (rs11568818) x MMP9 (rs17576) alleles showed a statistically significant association with an increased risk of EGE, while 10 other combinations correlated with a reduced risk of the disease. MDR analysis produced two statistically significant models for MMP allelic combinations involved in EGE progression, both with 100% penetrance and 83% and 78% accuracy.
Asunto(s)
Endometriosis/genética , Metaloproteinasa 3 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/genética , Alelos , Endometriosis/patología , Etnicidad/genética , Femenino , Estudios de Asociación Genética , Humanos , Polimorfismo de Nucleótido Simple , Federación de RusiaRESUMEN
Uterine leiomyoma (UL) is a benign and most common tumor that affects 20-45% of women of fertile age. In this study, we analyzed the MED12 second exon nucleotide sequence from 15 DNA samples extracted from LM of 15 subjects with uterine leiomyoma and 15 DNA samples extracted from peripheral blood leukocytes of the same female subjects. It was shown that somatic mutations in the MED12 gene occur in 73% of cases with deletions of varying sizes and missense mutations being most common at codon 44. Mutations in the MED12 gene could play an indirect role in leiomyoma progression by modifying the activity of other genes that encode proteins involved in growth and tumor progression.
Asunto(s)
Eliminación de Gen , Leiomioma/genética , Complejo Mediador/genética , Mutación Missense , Neoplasias Uterinas/genética , Adulto , Anciano , Femenino , Humanos , Persona de Mediana EdadRESUMEN
A novel phenomenon of unusual selective acridine orange (AO) staining ofpericentromeric heterochromatin regions (HRs) in chromosomal preparations from tissue with known spontaneous mitotic activity (chorionic villi, placenta, embryonic tissues, bone marrow, and testes), as well as embryonic stem cells, is described. Staining with 0.01% AO in a citric-phosphate (pH 5.5) or sodium phosphate (pH 7.0) buffer solution allows the HRs of human chromosomes (1q12, 9q12, 13p11.2, 14p11.2, 15p11.2, 16q11.2, 21p11.2, 22p11.2, and Yq12) and pericentromeric HRs of mouse chromosomes to be reliably detected by the red fluorescence of AO. This method of AO staining does not require any pretreatment. Explanations for metachromatic AO staining of polymorphic pericentromeric HRs in chromosomes of spontaneously dividing cells are suggested. A high reproducibility of the specific AO staining makes it possible to suggest its use as a reliable quick method for detection of polymorphic HRs of human chromosomes in cytogenetic prenatal diagnosis and oncohematology.
Asunto(s)
Naranja de Acridina/química , Cromosomas Humanos/genética , Heterocromatina , Polimorfismo Genético , Animales , Células de la Médula Ósea/citología , División Celular/genética , Corion/citología , Femenino , Heterocromatina/genética , Humanos , Células Madre Mesenquimatosas/citología , Metafase/genética , Ratones , Embarazo , Coloración y Etiquetado/métodosRESUMEN
Impressive advances in the studies of human genome, identification of mutant genes of hereditary diseases and candidate genes of many chronic multifactorial diseases (MFD) laid the foundation of molecular medicine. Its characteristic features, such as the focus on individual prophylactic care, give reason to consider it as personalized predictive medicine (PPM). The fundamental concept behind PPM comprises the notion of genetic passport and its methodological basis is genetic testing (GT). Recent progress in PPM has been achieved due to the introduction of comprehensive genomic screening of associations. At the same time, the contribution of known individual genes to the development of MFD appears to be relatively insignificant which does not allow to identify the main causes of MFD. It gave rise to some scepsis as regards the value of genome as a source of information for practical medicine. Possibilities for the improvement of GT and conditions for the introduction of the available data into clinical practice are discussed. The necessity to attract clinicians to the work on PPM is emphasized. The development of unified MFD gene panels for clinical application and software for the evaluation and interpretation of GT results for doctors and patients is an indispensable condition for the use of PPM knowledge in the healthcare practice. The importance of solution of relevant ethical, juridical, and social issues is underscored.
Asunto(s)
Predisposición Genética a la Enfermedad , Servicios Genéticos/normas , Pruebas Genéticas/normas , Herencia Multifactorial/genética , Medicina de Precisión , Privacidad Genética/ética , Servicios Genéticos/ética , Servicios Genéticos/tendencias , Pruebas Genéticas/ética , Pruebas Genéticas/tendencias , Genoma Humano , Humanos , Linaje , Medicina de Precisión/ética , Medicina de Precisión/normas , Medicina de Precisión/tendencias , Mejoramiento de la CalidadRESUMEN
We performed a stage-by-stage study of DNA methylation patterns in metaphase chromosomes from blastomeres of triploid and abnormal diploid human embryos. QFH-banded homologous parental chromosomes differ in their DNA methylation patterns at the metaphase of the 1st cleavage division. Chromosomes of both parental genomes are gradually demethylated at subsequent cleavages, undergoing hemimethylation in 2-cell embryos. At the 4-cell stage hypomethylated chromosomes initially appear and are further registered until the blastocyst stage. The proportion of hemimethylated and hypomethylated chromosomes varies between the blastomeres since the 4-cell stage with no preference for certain chromosomes to be hemi- or hypomethylated demonstrates random segregation of hypomethylated, undermethylated and methylated chromatids during cell cleavage. By the blastocyst stage the chromosomes acquire band- and, thus, chromosome-specific methylation patterns, with 5-methylcytosine-rich DNA preferentially accumulated in R- and T-bands and in the short arms of acrocentric chromosomes. Thus, demethylaton and remethylation of parental genomes of human embryos proceeds in the same manner from the 1st metaphase stage up to the blastocyst. These processes involve all chromosomes and all bands from each chromosome and lead to establishment of chromosome-specific DNA methylation patterns by the blastocyst stage with no differences between homologous chromosomes.
Asunto(s)
Blastocisto , Cromosomas Humanos , Metilación de ADN , Metafase , Humanos , Hibridación Fluorescente in SituRESUMEN
New molecular-genetic methods stimulate substantial advances in complex diseases studies, speed up identification of new candidate genes participating in functional genetic modules (gene nets) associated with many common diseases. Decisive impact of predictive genetic studies in efficient implementation of genomic technology advances into presymptomatic identification of the subjects of high risk groups prone to various common complex diseases is reviewed. Substantial progress of genomic studies in genetic of aging processes, including complex metabolic processes and gene regulation is outlined. Advances of predictive medicine in pharmacogenomic, nutrigenomic, sport genomic as well as in genomic of aging substantiate real and soon progress in promotion active healthy longevity.
Asunto(s)
Envejecimiento/genética , Envejecimiento/metabolismo , Genómica/tendencias , Senescencia Celular/genética , Radicales Libres/metabolismo , Estudio de Asociación del Genoma Completo , Genómica/métodos , Humanos , Insulina/metabolismo , Longevidad/genética , Estrés Oxidativo/genética , Transducción de Señal/genéticaRESUMEN
Densitometry of cosmonauts following long-duration missions shows reduction of bone mineral density (BMD). On the average, post-flight BMD remains within the normal range and the broad variability of individual BMD values sometimes is qualified as local osteopenia. Individual reactions are typed by similarity of amount and rate of BMD loss. At present, analysis of functionally significant polymorphism of bone metabolism genes is the most effective instrument for diagnostics of susceptibility to osteopenia and osteoporosis. The investigation was aimed to analyze polymorphism of genes of vitamin-D and (VDR) and calcitonin (CALCR) receptors, and of collagen-1 alpha-1-chain (Col1a-1) in candidate cosmonauts and cosmonauts returned from 5 to 7-mo. missions. According to the results of analysis, in the majority of cosmonauts rapid BMD loss correlated with TT genotype by VDR gene but not with genotypes Tt and tt and associated with carriage of incomplete s-allele in the Col1a1 gene. Yet, in several instances high BMD loss rates were personified with carriers of VDR gene alleles (homo- and heterozygote states--tt and Tt) and heterozygote by Col1a1 gene (Ss).
Asunto(s)
Densidad Ósea/genética , Enfermedades Óseas Metabólicas/genética , Colágeno Tipo I/genética , ADN/genética , Polimorfismo Genético , Receptores de Calcitonina/genética , Receptores de Calcitriol/genética , Astronautas , Enfermedades Óseas Metabólicas/metabolismo , Colágeno Tipo I/metabolismo , Cadena alfa 1 del Colágeno Tipo I , Estudios de Seguimiento , Humanos , Reacción en Cadena de la Polimerasa , Receptores de Calcitonina/metabolismo , Receptores de Calcitriol/metabolismo , Estudios Retrospectivos , Factores de RiesgoRESUMEN
[The multipurpose probe for real-time assessment of behavior reactions of marine mammals and concurrent temperature and noise pollution measurements was subjected to field testing]