A. Aránega

Regulatory systems in bone marrow for hematopoietic stem/progenitor cells mobilization and homing.

Regulation of hematopoietic stem cell release, migration, and homing from the bone marrow (BM) and of the mobilization pathway involves a complex interaction among adhesion molecules, cytokines, proteolytic enzymes, stromal cells, and hematopoietic cells. The identification of new mechanisms that regulate the trafficking of hematopoietic stem/progenitor cells (HSPCs) cells has important implications, not only for hematopoietic transplantation but also for cell therapies in regenerative medicine for patients with acute myocardial infarction, spinal cord injury, and stroke, among others. This paper reviews the regulation mechanisms underlying the homing and mobilization of BM hematopoietic stem/progenitor cells, investigating the following issues: (a) the role of different factors, such as stromal cell derived factor-1 (SDF-1), granulocyte colony-stimulating factor (G-CSF), and vascular cell adhesion molecule-1 (VCAM-1), among other ligands; (b) the stem cell count in peripheral blood and BM and influential factors; (c) the therapeutic utilization of this phenomenon in lesions in different tissues, examining the agents involved in HSPCs mobilization, such as the different forms of G-CSF, plerixafor, and natalizumab; and (d) the effects of this mobilization on BM-derived stem/progenitor cells in clinical trials of patients with different diseases.

Morphometric data concerning the great arterial trunks and their branches

In a total of 496 fetuses and newborns ranging in body weight from 60 to 5000 g, we performed a morphometric study of the vascular complex, that is the orifices of the great arterial trunks, ascending aorta, aortic arch, vessels branching from the aortic arch, the aortic isthmus, descending aorta, pulmonary trunk, orifices of the pulmonary arteries and arterial duct. In all, 17 different parameters were measured in each specimen, using several new parameters in addition to the measurements classically used in the aortic isthmus, arterial duct and pulmonary trunk. Anatomo-geometric models of these structures were developed, and stereometric formulas used to calculate the real volumes of the aortic isthmus, arterial duct and pulmonary trunk. The variations in the correlations between two measurable characteristics, that is, body weight and each of the morphometric parameters were analyzed, and the minimum, normal and maximum patterns of normality for each parameter were obtained with regression equations. The results show that the volume of the aortic isthmus increases by 0.03 ml for each millilitre increase in left ventricular volume. The isthmic volume increases by 0.32 ml for each millilitre increase in volume of the arterial duct and the volume of the pulmonary trunk increases by 6.4 ml for each increase of 1 millilitre in the volume of the aortic isthmus. The inner circumference of the aortic isthmus is greater than that of the arterial duct, whereas the former vessel is always shorter than the latter. We believe that these morphological data, when appropriately interpreted, have immediate clinical and surgical applications in the treatment of fetal and perinatal cardiovascular disorders.

Inverse expression of mdr 1 and c-myc genes in a rhabdomyosarcoma cell line resistant to actinomycin d.

Cytotoxic agents used in cancer therapy may induce differentiation in tumour cells with no proliferative potential. However, chemotherapy can also induce multidrug resistance, a formidable obstacle to the successful treatment of tumours. Both events were recently shown to occur in a rhabdomyosarcoma cell line (RD‐DAC) resistant to actinomycin D, a drug of choice in the treatment of these tumours. To analyse this connection, cell line RD cultures were investigated with progressive concentrations of actinomycin D and it was shown that a minimum dose (1·2×10−6 mm) of the drug was necessary to increase mdr 1 mRNA in RD‐DAC. The mechanism of mdr 1 overexpression was an increase in the number of copies of the mdr 1 gene, although the mRNA levels were not correlated with mdr 1 amplification. Drug resistance mediated by mdr 1 overexpression coincided with the development of myogenic differentiation in RD‐DAC and with a decrease in c‐myc mRNA levels, whereas levels of N‐myc mRNA showed no modulation. These findings suggest that factors implicated in cell proliferation and differentiation, such as c‐myc, may be responsible for the control of genes related to the development of multidrug resistance in rhabdomyosarcomas. Modulation of these factors may determine the sensitivity of rhabdomyosarcoma cells to drugs and may play an important role in triggering the differentiation programme found in these resistant rhabdomyosarcoma cells.

Morphometric data on the arterial duct in the human fetal heart

In a total of 496 fetuses and newborns ranging in body weight from 60 to 5000 g, we performed a morphometric study of the ascending aorta, the descending aorta, the aortic isthmus, the right pulmonary artery, the left pulmonary artery and the arterial duct. In all, nine different parameters were measured in each specimen. The variations in the correlations between two measurable characteristics, namely body weight and each of the morphometric parameters, were analyzed. The minimum, normal and maximum patterns of normality for each parameter were obtained with regression equations. We compared statistically the diameter of the arterial duct with the diameter of the ascending and descending aorta, the aortic isthmus, the right pulmonary artery and the left pulmonary artery with one way analysis of variance, using Bonferronis test in the pairwise comparisons. The diameter of the arterial duct was smaller than the diameter of the ascending and descending aortas, the aortic isthmus, the right pulmonary artery and the left pulmonary artery, and the diameter of the ascending aorta was larger than that of the descending aorta. These morphological data have immediate clinical and surgical applications in the treatment of fetal and perinatal cardiovascular disorders.

Colon Cancer Therapy: Recent Developments in Nanomedicine to Improve the Efficacy of Conventional Chemotherapeutic Drugs

The number of patients with colorectal cancer, the third most frequently diagnosed malignancy in the world, has increased markedly over the past 20 years and will continue to increase in the future. Despite recent advances in chemotherapy, currently used anticancer molecules are unable to improve the prognosis of advanced or recurrent colorectal cancer, which remains incurable. The transport of classical drugs by nanoparticles has shown great promise in terms of improving drug distribution and bioavailability, increasing tissue half-life and concentrating anticancer molecules in the tumor mass, providing optimal drug delivery to tumor tissue, and minimizing drug toxicity, including those effects associated with pharmaceutical excipients. In addition, colon cancer targeting may be improved by incorporating ligands for tumor-specific surface receptors. Similarly, nanoparticles may interact with key drug-resistance molecules to prevent a reduction in intracellular drug levels drug. Recently published data have provided convincing pre-clinical evidence regarding the potential of active-targeted nanotherapeutics in colon cancer therapy, although, unfortunately, only a few of these therapies have been translated into early-phase clinical trials. As nanotechnology promises to be a new strategy for improving the prognosis of colon cancer patients, it would be very useful to analyze recent progress in this field of research. This review discusses the current status of nanoparticle-mediated cancer-drug delivery, the challenges restricting its application, and the potential implications of its use in colon cancer therapy.

Modulation of α-Actin and α-Actinin Proteins in Cardiomyocytes by Retinoic Acid during Development

Early heart development is known to be sensitive to retinoid concentrations. Although the influence of retinoids on cardiac morphogenesis has been described previously, the effect of retinoids on cardiomyocyte differentiation during development has not been characterized. We quantified the effects of the retinoic acids all-trans RA and 13-cis RA on α-actin and α-actinin at the subcellular level in cultures of chick embryo cardiomyocytes obtained from Hamburger and Hamilton’s (HH) stage 22, 32 and 40 embryos. The retinoids increased the concentration of α-actin and α-actinin in the cytoplasmic and cytoskeletal fractions of cells at all three stages of development. The effect was greatest in cardiomyocytes treated for 24 h with all-trans RA and in cells from HH22 embryos. The greatest increases in α-actin concentration occurred in the cytoskeletal fraction of HH22 cells cultured for 24 h with all-trans or 13-cis RA, whereas the greatest increases in α-actinin were found in the cytoplasmic fraction of HH22 cells exposed to retinoids for 24 h. We conclude that retinoic acid plays a role in the reorganization of the pattern of sarcomeric protein expression during cardiomyocyte differentiation.

Development of Chick Cardiomyocytes: Modulation of Intermediate Filaments by Basic Fibroblast and Platelet-Derived Growth Factors

Recent studies suggest that peptide growth factors play a functional role in cardiac muscle. To test whether embryonic cardiac muscle is a target for regulation by basic fibroblast growth factor and platelet-derived growth factor, we analyzed the effects of these peptides on the expression of the intermediate filaments desmin and vimentin at the subcellular level during development. Sodium dodecyl sulfate-gel electrophoresis, immunoblotting and fluorescence-activated cell sorting analysis were used to study the effect of basic fibroblast growth factor and platelet-derived growth factor on cultures of chick cardiomyocytes during development. Cytoplasmic and cytoskeletal concentrations of desmin and vimentin were dependent on the stage of embryonic development and on the type of growth factor added to the culture. The most significant finding was the increase in desmin expression in the cytoplasmic and cytoskeletal compartments after treatment with basic fibroblast growth factor (10 ng/ml) of chick heart cells at Hamburger and Hamilton stage 19. In more mature stages, basic fibroblast growth factor did not modify the levels of desmin expression. However, this factor led to a progressive deceleration in the rate of increase in vimentin expression. Platelet-derived growth factor increased vimentin expression in all stages studied, the greatest increases appearing in early stages of heart development. Our findings support the hypothesis that basic fibroblast growth factor plays a role in cardiomyocyte differentiation during the early stages of development, whereas platelet-derived growth factor has a dedifferentiating effect.

Circulating α-actin protein in acute myocardial infarction

Abstract We used Western-blot analysis to investigate the possible presence in the bloodstream of the contractile protein α-actin in 70 patients diagnosed with acute myocardial infarction on the basis of clinical, electrocardiographic and laboratory (creatine kinase and lactate dehydrogenase) criteria. Circulating protein was identified with a monoclonal antibody specific for cardiac α-actin. Of the 70 control samples of blood, the immunoblot results were negative for α-actin in 98% of the cases. Of the 30 patients with skeletal muscle damage caused by surgery, 26 were negative for circulating α-actin. Of the 70 patients with acute myocardial infarction, circulating α-actin was found in 67 (95%) as a 43 kDa band in immunoblots; the highest circulating concentrations (0.0580 μg/μl) were found in those with anterior acute myocardial infarction. Circulating α-actin was detected in samples taken between 1 and 180 h after the onset of pain, and showed a biphasic pattern of appearance. Our findings for serum α-actin, together with the relationship between serum concentrations of this protein and sex ( p = 0.001), tobacco use ( p = 0.007) and postepisode complications ( p = 0.002), should make it possible to gain a deeper understanding of acute myocardial infarction as a clinical entity.

How is gene transfection able to improve current chemotherapy? The role of combined therapy in cancer treatment.

Despite advances in cancer treatment, a large number of patients eventually develop metastatic disease that is generally incurable. Systemic chemotherapy remains the standard treatment for these patients. Several chemotherapeutic combinations have proven effective in the management of cancer. Paradoxically, although the purpose of polychemotherapy is to improve the prognosis and prolong the survival of patients, it often carries considerable toxicity that causes substantial adverse symptoms. For this reason, a major goal of cancer research is to improve the effectiveness of these cytotoxic agents and reduce their adverse effects. Gene transfer has been proposed as a new strategy to enhance the efficacy of anti-tumor drugs in the treatment of intractable or metastatic cancers. In fact, the association of gene therapy and drugs (combined therapy) has been reported to increase the anti-proliferative effect of classical treatments in lung, bladder, pancreatic, colorectal and breast cancers, among others. Various especially promising therapies have been proposed in this context, including the use of suicide genes, antisense oligonucleotides, ribozymes and RNA interference. In this chapter, we review recent progress in the development of novel anti-cancer strategies that associate cytotoxic agents with gene transfer to enhance their antitumor effect.

Coexpression of intermediate filament proteins in the chick embryo heart.

We studied the distribution of intermediate filament proteins during several stages of chick embryo heart development by indirect immunofluorescence and fluorescence-activated cell surface analysis. Vimentin is the predominant intermediate filament during the early stages of cardiac genesis, while desmin appears essentially with maturation. Desmin is the main subunit protein of intermediate filaments in the mature myocyte.