Insulin-producing cells derived from stem cells: recent progress and future directions

Type 1 diabetes is characterized by the selective destruction of pancreatic beta-cells caused by an autoimmune attack. Type 2 diabetes is a more complex pathology which, in addition to beta-cell loss caused by apoptotic programs, includes beta-cell dedifferentiation and peripheric insulin resistance. beta-Cells are responsible for insulin production, storage and secretion in accordance to the demanding concentrations of glucose and fatty acids. The absence of insulin results in death and therefore diabetic patients require daily injections of the hormone for survival. However, they cannot avoid the appearance of secondary complications affecting the peripheral nerves as well as the eyes, kidneys and cardiovascular system. These afflictions are caused by the fact that external insulin injection does not mimic the tight control that pancreatic-derived insulin secretion exerts on the body's glycemia. Restoration of damaged beta-cells by transplantation from exogenous sources or by endocrine pancreas regeneration would be ideal therapeutic options. In this context, stem cells of both embryonic and adult origin (including beta-cell/islet progenitors) offer some interesting alternatives, taking into account the recent data indicating that these cells could be the building blocks from which insulin secreting cells could be generated in vitro under appropriate culture conditions. Although in many cases insulin-producing cells derived from stem cells have been shown to reverse experimentally induced diabetes in animal models, several concerns need to be solved before finding a definite medical application. These refer mainly to the obtainment of a cell population as similar as possible to pancreatic beta-cells, and to the problems related with the immune compatibility and tumor formation. This review will summarize the different approaches that have been used to obtain insulin-producing cells from embryonic and adult stem cells, and the main problems that hamper the clinical applications of this technology.     

Developmental stage of failed eggs in the red-legged partridge <Emphasis Type="Italic">Alectoris rufa</Emphasis>

Understanding whether egg failure occurs more frequently during laying or during incubation is important for determining the most vulnerable period of the egg phase in bird species with long laying and incubation periods. However, most studies compare nest failure during the egg stage and the hatchling stage. In this study we examined whether egg failure was higher during the laying period and early development than during the final stages of development in a population of red-legged partridge (Alectoris rufa) from northeast Spain. We collected 216 abandoned eggs in the field. Only 12% (n = 26) of the eggs were sufficiently intact to enable direct assessment of developmental stage, showing that 81% of 26 eggs were lost during early stages. The majority of eggs were depredated (n = 154, 71%), as evidenced by an opening hole. To take advantage of information contained in these eggs, we propose a simple method to assess their developmental stage by measuring the size of the opening of their hole. Based on this method, we estimated that the proportion of eggs lost during early stages ranged from 72 to 83%. We validated this result of a high percentage of egg failure during early developmental stages with a larger experimental sample. Nest losses in the partridge are higher during the laying period or early development. This result is important in that the direction of conservation efforts and their seasonal application should be adjusted to account for the most important factors affecting egg failure.     

Cytogenetics and karyosystematics of South American oryzomyine rodents (Cricetidae: Sigmodontinae). IV. Karyotypes of Venezuelan, Trinidadian, and Argentinian water rats of the genus Nectomys.

Bone marrow chromosomes were studied in South American water rats of the genus Nectomys from Venezuela, Trinidad, and Argentina. Specimens of N. squamipes from western and southern Venezuela showed a 2n = 52-53 karyotype, whereas a 2n = 56-57 karyotype was found in specimens from northeastern Argentina. In both cases, odd karyotypes can be explained by the presence of a supernumerary chromosome. In contrast, water rats from northeastern Venezuela and Trinidad showed a strikingly reduced 2n = 16-17 polymorphic chromosome complement. Six different karyomorphs were found among the latter, which may have resulted from a combination of pericentric inversions in two pairs of autosomes and a centromeric fusion in another autosomal pair. It is proposed that the new 2n = 16-17 cytotypes belong to a species of its own, for which the name N. palmipes is suggested.     

Partial characterization of neuropathy target esterase and related phenyl valerate esterases from bovine adrenal medulla

The mechanism by which organo-phosphorus-induced delayed polyneuropathy is induced relates to the specific inhibition and subsequent modification (“aging”) of a protein known as neuropathy target esterase (NTE), operatively defined as paraoxon-resistant and mipafox-sensi-tive phenyl valerate (PV) esterase activity. This protein has fundamentally been investigated in hen brain, the latter being the habitually employed OPIDP study model. In the present article, a partial characterization is made of the NTE and other related PV esterases in the bovine adrenal medulla and brain; NTE sensitivity to the neurotoxic or-ganophosphorus compound mipafox is investigated, and its subcellular distribution is studied. The NTE activity of the adrenal medulla was found to be the highest of those among the tissues studied to date (5000 ± 1400 mU/g tissue; ± SD, n = 12). This activity represented 93% of the PV esterase activity resistant to 40 μm paraoxon in the par-ticulate fraction of the adrenal medulla and approximately 50% of total PV esterase activity. In the bovine brain, these proportions were 72 and 26%, respectively, i.e., similar to those described in hen brain. The mipafox inhibition curve of PV esterase activity resistant to 40μM paraoxon in the particulate fraction of the adrenal medulla suggests that NTE activity fundamentally comprises a mipafox-sensitive component with an I ₅₀ of 6.39 μM at 30 minutes, which is similar to the value reported in hen brain. NTE activity in the bovine adrenal medulla is almost exclusively limited to the particulate fraction, the microsomal fraction, plasma membrane, and chromaffin granule-enriched fractions being the highest in terms of specific activity. On the contrary, the mitochondria-enriched fraction was very poor in such activity. In bovine brain, most NTE activity was likewise limited to the particulate fraction.     

Phenotypic and functional characterization of glucagon-positive cells derived from spontaneous differentiation of D3-mouse embryonic stem cells

BackgroundGlucagon expression is being considered as a definitive endoderm marker in protocols aiming to obtain insulin-secreting cells from embryonic stem cells. However, it should be considered that in vivo glucagon is expressed both in definitive endoderm- and neuroectoderm-derived cells. Therefore, the true nature and function of in vitro spontaneously differentiated glucagon-positive cells remains to be established.MethodsD3 and R1 mouse embryonic stem cells as well as α-TC1-9 cells were cultured and glucagon expression was determined by real-time PCR and immunocytochemistry. Functional analyses regarding intracellular calcium oscillations were performed to further characterize glucagon⁺ cells.ResultsSpecifically, 5% of D3 and R1 cells expressed preproglucagon, with a small percentage of these (<1%) expressing glucagon-like peptide 1. The constitutive expression of protein convertase 5 supports the expression of both peptides. Glucagon⁺ cells co-expressed neurofilament middle and some glucagon-like peptide-1⁺ cells, glial fibrillary acidic protein, indicating a neuroectodermic origin. However, few glucagon-like peptide-1⁺ cells did not show coexpression with glial fibrillary acidic protein, suggesting a non-neuroectodermic origin for these cells. Finally, glucagon⁺ cells did not display Ca²⁺ oscillations typical of pancreatic α-cells.DiscussionThese results indicate the possible nondefinitive endodermal origin of glucagon-positive cells spontaneously differentiated from D3 and R1 cell lines, as well as the presence of cells expressing glucagon-like peptide-1 from two different origins.     

A proposed unified nomenclature for the enamelled components of the molar teeth of the Cricetidae (Rodentia)

The primitive topography of the enamelled surface of molar teeth of the Cricetidae is described as expressed in the fossil Cricetodontinae. Morphological variations in the molar structure of different subgroups among the Cricetidae are interpreted as derivations from this cricetodontine pattern. Eleven available naming systems for such components are surveyed, and a new unifying nomenclature is proposed, based on the Cope-Osborn cusp homologies for mammals. Names for enamel cusps, cuspules, styles, lophs, folds and islands are given, in an attempt to include in an overall general nomenclature the advantages of the most valuable, already available, nomenclatorial systems. The system purports to apply to all modifications of the cricetid crown molar pattern, and it claims to fulfil the need for a uniform nomenclature.     

Synthesis and biological activity of substance P analogues

The synthesis of the hexapeptide [Glu6]SP6-11 and its glycosylated analogue at the Glu6 gamma-carboxyl position by solution procedures according to several strategies is discussed. The biological activity of SP, [Glu6]SP6-11 (VI) and [Glu(beta-D-Glcp)6]SP6-11 (VIII) have been determined and compared to SP by the GPI and RVD assays. The introduction of a beta-D-glucopyranosyl moiety at the sixth position of the [Glu6]SP6-11 did not affect to a great extent the in vitro activity pattern of the parent hexapeptide.