El cultivo celular en la investigación básica del cáncer de mama

Resumen La técnica del cultivo celular es la que ha permitido conocer el comportamientoin vitro de las células cancerosas. En esta revisión pretendemos introducir las peculiaridades básicas del cultivo celular, referido especialmente a líneas cancerosas mamarias, relacionar el origen de las líneas celulares más utilizadas en la investigación de este cáncer, mencionar las técnicas de laboratorio que pueden aplicarse sobre estos cultivos y ejemplificar la utilidad de las mismas, tomando como modelo diversos trabajos que estudian los efectos del factor de crecimiento epidérmico sobre líneas celulares hormonoindependientes de cáncer de mama.      

Resolution of peripheral artery catheter-induced ischemic injury following prolonged treatment with topical nitroglycerin ointment in a newborn: a case report.

Tissue ischemia, necrosis, and gangrene are uncommon but well-described complications of arterial catheterization in the neonate. Treatment options for progressive tissue necrosis following arterial embolization and/or vasospasm are limited in these patients secondary to unpredictable pharmacokinetics and risks associated with systemic anticoagulation or vasodilatation in newborns. We report a case of a multidose regimen of topical 2% nitroglycerin ointment for reversing severe tissue ischemia following peripheral arterial line placement. The favorable response in this infant suggests that topical nitroglycerin therapy should be considered as potential therapy to ameliorate the effects of vascular compromise following arterial line placement in neonates.     

Ischemic stroke complicating cardiac catherization: case report.

Ischemic stroke occurs in 0.2-0.4% of patients undergoing left heart catheterization, and is responsible for 5-10% of the mortality associated with the procedure. The main predisposing factors for this complication are female gender, complex atherosclerotic plaques in the ascending aorta, and peripheral arterial disease. The possibility of timely intervention with reperfusion therapy supports close clinical monitoring during the immediate post-catheterization period. The cardiologist should be familiar with the various types of stroke reperfusion therapy and its indications according to the time interval between catheterization and the stroke. The decision should be discussed with neurology and neuroradiology.     

Benefits and risks of simvastatin in patients with familial hypercholesterolaemia.

Familial hypercholesterolaemia is a frequent, inherited, monogenic disorder, associated with accelerated development of atherosclerotic disease leading to coronary artery disease. Life expectancy of patients with familial hypercholesterolaemia is reduced by 15-30 years unless they are adequately treated with lipid-lowering therapy. Given the chronic nature of this disease, the selection of a therapeutic approach should be strongly based on its long-term safety and tolerability. The introduction of HMG-CoA reductase inhibitors has revolutionised the treatment of familial hypercholesterolaemia.Simvastatin 40-80 mg/day effectively reduces serum low density lipoprotein (LDL)-cholesterol levels. Furthermore, simvastatin reduces triglycerides and mildly raises high density lipoprotein-cholesterol levels. In addition to the hypolipidaemic effect, other potentially important effects, such as improvement of endothelial function and reduction of LDL oxidation and vascular inflammation, have been associated with HMG-CoA reductase inhibitor therapy. Simvastatin has also been shown to abolish the progression, and even facilitate the regression, of existing human atherosclerotic lesions.The good safety and tolerability profile of simvastatin is clearly highlighted by the low rate of therapy discontinuation observed in several population-based clinical trials. The most common adverse events leading to the discontinuation of therapy are gastrointestinal upset and headache. Asymptomatic elevations in liver transaminase levels and myopathy are uncommon.The overwhelming clinical evidence regarding the long-term use of HMG-CoA reductase inhibitor therapy in patients with familial hypercholesterolaemia together with the long-term safety data (particularly relating to simvastatin) provide support for the use of this drug as a first-line agent when pharmacological treatment is indicated. Early intervention with simvastatin treatment can be successfully implemented with favourable economic benefits.     

CD24 expression on human keratinocytes

Abstract: CD24 or Nectadrin is a cell surface glycoprotein expressed in pre-B lymphocytes, T lymphocytes, neurons, muscle cells and carcinoma cells. Its function is not completely known, but it has been suggested that it is involved in cell adhesion and signalling. CD24 has recently been identified as the human molecule homologous to the murine heat-stable antigen (HSA). HSA is expressed by murine keratinocytes and delivers costimulatory signals in T-cell activation. Long-term cultures of normal human keratinocytes (HKC) were obtained from skin of human female breast sections and either left untreated or were treated with phorbol-12-myristate-13-acetate (PMA) at 10–100 ng/ml, calcium 0.5–2 mM or IFN-γ 100–1000 U/ml, for 24–48 h. Using RT-PCR and flow cytometry we showed that HKC express low levels of CD24 even under basal conditions, and the treatment with calcium, PMA or IFN-γ increased levels of CD24 mRNA and protein. To the best of our knowledge, this is the first report to measure CD24 expression and production by cultured HKC in basal conditions and after stimulation. Further studies are needed to determine biological and therapeutical relevance of these findings.     

Classification of thoracic and lumbar spine fractures: problems of reproducibility

Reproducibility of fracture classification systems in general has been a matter of controversy. The reproducibility of spinal fracture classifications has not been sufficiently studied. We studied the inter-observer and intra-observer reproducibility of the Magerl (AO) classification using radiograms, CTs and MRIs of 53 patients. We compared this classification with the older and simpler Denis classification. Five observers classified the fractures, first using the radiograms and CTs and, 6 weeks later, with radiograms and MRIs. Three of the observers repeated the readings after 3 months. Three observers also classified the fractures according to Denis. Agreement was measured using Cohen's kappa test. The type (A, B, C) classification of the AO system was fairly reproducible with CTs. With MRI this was only moderate. Group subclassification of the types yielded higher kappa values, corresponding to substantial agreement. The agreement was, in general, better with the Denis classification, but the variance was higher due to the difficulty of finding proper categories for some injury patterns. Although the AO classification allows proper registration of all kinds of injury, the reproducibility, especially at the type level, is problematic. Use of MRI and better definition of the distinctive properties of the three different types may enhance the reproducibility of the scheme.     

Differential effect of taurine and serotonin on the outgrowth from explants or isolated cells of the retina

The sulfur amino acid taurine and the indoleamine serotonin increases and decreases, respectively, the outgrowth from goldfish retinal explants. Taurine seems to be acting, at least partially, through an increase in calcium fluxes, and the serotonin-inhibiting effect appears to be mediated by serotonin_1A receptors and cAMP. Isolated cells of postcrush goldfish retina and of retina from 5-day-old rats were cultured in the presence of taurine or serotonin. In the goldfish, the classical morphology of postcrush ganglion cells was observed. An antibody against the glycoprotein Thy-1 labelled three types of cells in the cultures of goldfish retina. The number of cells outgrowing and the length of the main neurite was measured at 5 days in culture in both species. The number of cells presenting neurites was increased in the goldfish retina by the addition of taurine, and decreased by serotonin. However, the length of the neurites was unaffected by the addition of the modulators. In the rat, only a slight decrease in the number of cells outgrowing was observed in the presence of serotonin. The incorporation of [³H]thymidinewas not modified after 5 days in culture in the presence of taurine or serotonin, either in the goldfish or in the rat retina. The antibody Thy 1.1 can label retinal cells of the goldfish invitro, one of them being ganglion cells. The trophic effect exerted by taurine in the postcrush goldfish retina needs the integrity of the tissue favoring the interaction of cells and factors, because outgrowth increases in retinal explants, but not in isolated cells.     

Effect of elevated prolactin levels on the synthesis and release of catecholamines from the adrenal medulla in female rats

Recent evidence suggests that increased plasma prolactin (PRL) levels could be modifying the synthesis and release of catecholamines (CA) from the adrenal medulla. In order to study this possibility, female rats bearing an anterior pituitary gland, from a litter-mate donor, under the right kidney capsule since day 30 of life and their sham-operated controls were sacrificed by decapitation 45 days after the transplant operation. Plasma PRL and CA levels and CA content in the adrenal medulla were analyzed. A significant increase in plasma PRL levels was shown in grafted (22 +/- 2 micrograms/l) when compared to control rats (6 +/- 0.3 micrograms/l), together with a significant increase in plasma norepinephrine (NE) (4.2 +/- 1.1 micrograms/l vs. 1.7 +/- 0.7 micrograms/l) and epinephrine (E) (2.9 +/- 0.6 micrograms/l vs. 1.6 +/- 0.2 micrograms/l). Similar plasma dopamine (DA) levels were found in both groups (0.6 +/- 0.2 micrograms/l vs. 0.8 +/- 0.3 micrograms/l). An increase in adrenal DA content (29.1 +/- 6.8 ng/mg protein), together with a decrease in NE (33.3 +/- 6.8 ng/mg protein) and E content (65.9 +/- 11.8 ng/mg protein) was detected in grafted as compared to control rats (DA: 12.0 +/- 3.6 ng/mg protein; NE: 79.3 +/- 22.1 ng/mg protein; E: 184.2 +/- 39.2 ng/mg protein). We can conclude from these data that the elevation of circulating PRL levels induced by a pituitary graft is able to increase the synthesis and release of CA from the adrenal medulla in female rats.     

MR susceptometry: an external-phantom method for measuring bulk susceptibility from field-echo phase reconstruction maps

A novel method for estimating the susceptibility of an object by using the magnetic resonance (MR) imaging field distortions in an external-reference water bath next to the object is described. The field measurement was obtained with a phase reconstruction from a gradient-echo acquisition. A field model of an arbitrary object in a static magnetic field was discretely calculated from geometry determined from the magnitude reconstruction. Least-squares estimation yields the susceptibility of the object. Required (and proven) assumptions include validity of superposition, object homogeneity, negligible higher-order field terms, field-model accuracy, geometric-model accuracy, and correlation of gradient-echo phase to field distortion. MR susceptometry estimation of phantoms yielded susceptibility estimates that correlated well with known values (r >.9975). This MR susceptometry method is a first step toward quantitation of iron levels through MR imaging phase information in patients with iron overload.