Lupine as a potential agent against diet-induced obesity through adenosine monophosphate-activated protein kinase pathway: crude oil versus nanoemulsion formulations

Obesity represents a major challenge to the pharmaceutical community due to the minimal availability of anti-obesity drugs and the drawbacks of current weight-loss agents. The study described herein presents lupine oil, in two pharmaceutical formulations, as a potential anti-obesity agent via its effect on different physiological, biochemical, and hormonal parameters. Rats were divided into two groups; one group was continued on a standard commercial rodent diet and served as the non-obese control. The other group was fed a high-fat diet for 7 weeks to prepare an obese rat model. Then, the obese rats were divided into groups to receive 100 mg/kg of the crude lupine oil or nanoemulsion for 10 or 20 days. Lupine oil showed a potent body weight-reducing effect and improved insulin resistance. The oil altered obesity-induced hyperlipidemia and it enhanced the leptin/adiponectin/AMPK hormonal system in epididymal fat, serum, and liver, to which all the above physiological activities could be attributed. The nanoemulsion formulation of lupine oil significantly amplified the activity for all the above physiological and hormonal parameters when compared to the crude oil formulation. Lupine oil nanoemulsion could be used as a potential drug against diet-induced obesity.

Neurodegeneración con acúmulo cerebral de hierro / Neurodegeneration with brain iron accumulation

Introducción. Las alteraciones en el metabolismo del hierro se han relacionado con diversas enfermedades neurodegenerativas, en algunas de ellas se trata del principal elemento de la enfermedad mientras que en otras, como la enfermedad de Alzheimer o Parkinson, se ha descrito su alteración pero no se conoce su significado fisiopatológico exacto. Objetivo. Revisión bibliográfica sobre el conocimiento actual de las enfermedades que cursan con depósitos encefálicos de hierro. Desarrollo. Se denomina neurodegeneración asociada a depósito de hierro (NAEH), a un grupo heterogéneo de enfermedades hereditarias (la mayor parte de ellas autosómicas recesivas) que cursan con depósitos de hierro en determinadas áreas cerebrales. La causa más frecuente es la mutación en el gen PANK2, recibiendo el nombre de Neurodegeneración asociada a Pantotenatocinasa (NAPC). La descripción inicial corresponde al síndrome de Hallervorden-Spatz. El cuadro clínico incluye manifestaciones motoras y deterioro psicomotor con una evolución progresiva hasta la incapacidad y fallecimiento del paciente. La resonancia magnética muestra alteraciones características afectando fundamentalmente al globo pálido. También se incluyen dentro de la NAEH la Neuroferritinopatia, la aceruloplasminemia y algunas formas asociadas a mutaciones en el gen PLA2G6, enfermedades que comparten algunas características clínicas y la presencia de alteraciones en la RM aunque con diferencias entre ellas. Conclusiones. El conocimiento de las distintas alteraciones genéticas ha permitido una mejor clasificación nosológica de la NAEH. Tanto la clínica como el patrón de alteraciones en RM pueden resultar de utilidad en el diagnóstico. Por el momento no se dispone de tratamientos que modifiquen el curso de la enfermedad (AU)

Introduction. Alterations in iron metabolism have been related with several neurodegenerative diseases. In some cases it is the main element of the disease while in others, such as Alzheimer's or Parkinson’s disease, its alteration has been reported but its exact pathophysiological significance remains unknown. Aims. The aim of the study was to carry out a review of the literature on the current knowledge about diseases that are accompanied by deposits of iron in the brain. Development. The term neurodegeneration with brain iron accumulation (NBIA) is used to refer to a heterogeneous group of hereditary (mostly autosomal recessive) diseases that are accompanied by deposits of iron in certain areas of the brain. The most frequent cause is a mutation in the PANK2 gene, which is called Pantothenate Kinase-Associated Neurodegeneration (PKAN). The initial description corresponds to Hallervorden-Spatz syndrome. The clinical picture includes motor manifestations and psychomotor deterioration with a progressive development until disability and the death of the patient. Magnetic resonance imaging shows characteristic alterations mainly involving the globus pallidus. NBIA also includes neuroferritinopathy, aceruloplasminemia and some forms associated to mutations in the PLA2G6 gene, diseases that share certain clinical characteristics, and the presence of alterations in the MRI scan, although there are differences from one to another. Conclusions. Understanding the different genetic alterations has made it possible to achieve a better nosological classification of NBIA. Both the clinical features and the pattern of alterations in MRI can be useful in the diagnosis. For the time being there are no treatments that modify the course of the disease (AU)

Targeted covalent drugs of the kinase family.

In the past decade tremendous progress has been made toward a new class of therapeutics termed 'targeted covalent drugs', in which structure-based approaches are employed to create small molecules that inactivate their protein target through targeted covalent attachment to a specific cysteine. In the kinase field, this approach is demonstrating promise in overcoming the potency, selectivity, and efficacy challenges currently faced by reversible kinase inhibitors, with several advancing into late stage clinical testing. This design paradigm has been successfully applied to making drug candidates for epidermal growth factor receptor (EGFR), Her2, and Bruton's tyrosine kinase (Btk). Here we review recent pre-clinical and clinical advances with targeted covalent kinase inhibitors, and the potential for broader application of the approach.

Microinjection strategies for the study of mitogenic signaling in mammalian cells.

First used in the analysis of dynamic changes in cell structure, microneedle microinjection allows in situ study of individual living cells as opposed to large scale metabolic analysis of heterogeneous cell culture. In addition, microinjection also offers the possibility to examine in vivo regulated processes by modulating the intracellular levels and activity of key regulatory proteins and genes in both a specific and controlled manner. A number of different strategies have been developed over the past 5 years to examine the pathways and effectors that are involved in mitogenic signaling as well as in the regulation of gene expression during the proliferative response to growth factors by normal fibroblasts. These strategies include: 1. Direct in vivo competition for various trans-activating DNA binding activities by microinjection of double-stranded oligonucleotides, microinjection of monospecific antibodies against transcription factors and microinjection of dominant negative mutants of transcription factors based upon their DNA binding domain. 2. Microinjection of purified enzymes (kinases and phosphatases) or peptides and antibodies that specifically inhibit these activities. 3. Microinjection of expression plasmids which encode various normal and epitope-tagged regulatory molecules. In many of the experiments described below, c-fos gene expression was monitored as an early marker of mitogenic response. The c-fos gene belongs to a family of genes whose transcription is activated very early after addition of growth factor (1-4). For in vivo studies, the c-fos promoter offers several unique advantages. Primarily, it is easy to manipulate. In practical terms, when mammalian fibroblasts are made quiescent (by replacing the normal growth media, with growth factors-depleted media) and subsequently activated by re-adding mitogen (growth factors, serum), c-fos RNA expression is restored within 15 minutes and the protein is specifically detected in the nuclei of cells after 90 minutes, but is no longer detectable after 3 hours. Secondly, results obtained with the c-fos promoter are directly applicable to cell growth since expression of c-fos is itself a prerequisite for proliferation as demonstrated by microinjection of anti-fos antibodies which prevented proliferation in mammalian cells (5). Thirdly, the c-fos promoter is exquisitely sensitive to agents which cause cell stress. In this respect, heat-shock, poor microinjection or microinjection in the presence of heavy metals or chelating agents in the culture media all rapidly stimulate c-fos expression. However, when compared to c-fos expression in the proliferative response, stress mediated c-fos expression is induced both more rapidly and strongly, reverses more slowly (the protein is still detectable after 5-6 hours) and does not result in cell proliferation (unpublished observation). As such, it provides an excellent internal control for identifying poor treatment and manipulation of cells . Finally, the c-fos promoter is subject to several levels of auto-regulation enabling the analysis of not only components involved in transcriptional activation , but also various aspects of transcriptional down regulation and shut-off.