Incidence, recurrence and cost of hyperglycaemic crises requiring emergency treatment in Andalusia, Spain.

AIMS: Hyperglycaemic crises (diabetic ketoacidosis and hyperosmolar hyperglycaemic state) are medical emergencies in people with diabetes. We aimed to determine their incidence, recurrence and economic impact. METHODS: An observational study of hyperglycaemic crises cases using the database maintained by the out-of-hospital emergency service, the Healthcare Emergency Public Service (EPES) during 2012. The EPES provides emergency medical services to the total population of Andalusia, Spain (8.5 million inhabitants) and records data on the incidence, resource utilization and cost of out-of-hospital medical care. Direct costs were estimated using public prices for health services updated to 2012. RESULTS: Among 1 137 738 emergency calls requesting medical assistance, 3157 were diagnosed with hyperglycaemic crises by an emergency coordinator, representing 2.9 cases per 1000 persons with diabetes [95% confidence intervals (CI) 2.8 to 3.0]. The incidence of diabetic ketoacidosis was 2.5 cases per 1000 persons with diabetes (95% CI 2.4 to 2.6) and the incidence of hyperosmolar hyperglycaemic state was 0.4 cases per 1000 persons with diabetes (95% CI 0.4 to 0.5). In total, 17.7% (n = 440) of people had one or more hyperglycaemic crisis. The estimated total direct cost was €4 662 151, with a mean direct cost per episode of €1476.8 ± 217.8. CONCLUSIONS: Hyperglycaemic crises require high resource utilization of emergency medical services and have a significant economic impact on the health system.

Influence of HLA class I, HLA class II and KIRs on vertical transmission and chronicity of hepatitis C virus in children.

BACKGROUND & AIM: There is evidence that maternal viral load of HCV during delivery influences the risk for Mother-to-child transmission (MTCT), but this does not explain all cases. We study the role of the immunogenetic profile (HLA, KIRs and KIR-ligand binding) of mothers and children in HCV-MTCT and in chronicity in the children. METHODOLOGY: 79 HCV-RNA (+) mothers and their 98 children were included. 24 children were infected, becoming chronic in 8 cases and clearing in 16. HLA-class-I and II and KIRs were determined by Luminex. RESULTS: MTCT study: The presence of HLA-C1-ligand in mothers and/or their children reduces the risk of transmission (mothers: Pc = 0.011, children: P = 0.033), whereas the presence of HLA-C2C2-ligand in mothers increases it (Pc = 0.011). In children KIR2DL3-HLA-C1 is a protector factor (Pc = 0.011). Chronicity in children study: Maternal DQA1*01 allele (Pc = 0.027), KIR2DS1 (Pc = 0.011) or KIR3DS1 (Pc = 0.011) favours chronicity in the child. The presence of the DQB1*03 allele (Pc = 0.027) and KIR2DS3 (P = 0.056) in the child and homozygosity for KIR3DL1/3DL1 (Pc = 0.011) and for the HLA-Bw4/Bw4 ligand (P = 0.027) is associated with viral clearance, whereas the presence of HLA-Bw6 ligand (P = 0.027), the binding of KIR3DS1-HLA-Bw4 (P = 0.037) and heterozygosity for KIR3DL1/3DS1 (Pc = 0.011) favour viral chronicity. Mother/child allele matching: In the joint HLA analysis, matching was greater between mothers and children with chronic infection vs those who had cleared the virus (67%±4.1 vs 57%±1.2, P = 0.003). CONCLUSIONS: The HLA-C1 ligand in the mother is related to MTCT, while several genetic factors of the mother or child are involved in the chronification or clearance of infection in the child. Matching allelic data is considered to be an indicator of HCV chronicity in the child and can be used as a potential prognostic test. This implies that NK cells may play a previously undocumented role in protecting against MTCT and that both NK cell immunity and adaptive T-cell responses may influence viral clearance in infected children.

Functionalized magnetic nanoparticles as vehicles for the delivery of the antitumor drug gemcitabine to tumor cells. Physicochemical in vitro evaluation.

Gemcitabine is a chemotherapy drug used in different carcinomas, although because it displays a short biological half-life, its plasmatic levels can quickly drop below the effective threshold. Nanoparticle-based drug delivery systems can provide an alternative approach for regulating the bioavailability of this and most other anticancer drugs. In this work we describe a new model of composite nanoparticles consisting of a core of magnetite nanoparticles, coated with successive layers of high molecular weight poly(acrylic acid) and chitosan, and a final layer of folic acid. The possibility of using these self-assembled nanostructures for gemcitabine vehiculization is explored. First, the surface charge of the composite particles is studied by means of electrophoretic mobility measurements as a function of pH for poly(acrylic acid) (carbopol) of different molecular weights. The adsorption of folic acid, aimed at increasing the chances of the particles to pass the cell membrane, is followed up by optical absorbance measurements, which were also employed for drug adsorption determinations. As a main result, it is shown that gemcitabine adsorbs onto the surface of chitosan/carbopol-coated magnetite nanoparticles. In vitro experiments show that the functionalized magnetic nanoparticles are able to deliver the drug to the nuclei of liver, colon and breast tumor cells.

Nanoengineering of doxorubicin delivery systems with functionalized maghemite nanoparticles.

Superparamagnetic iron oxide nanoparticles are developing as promising candidates for biomedical applications such as targeted drug delivery. In particular, they represent an alternative to existing antitumor drug carriers, because of their ultra-fine size, low toxicity and magnetic characteristics. Nevertheless, there is a need to functionalize them in order to achieve good biocompatibility, efficient modification for further attachment of biomolecules, and improved stability. In this work we describe the functionalization of superparamagnetic maghemite nanoparticles encapsulated in a silica shell. After their chemical modification with positive (3-aminopropyl)trimethoxysilane, a gold layer was deposited in order to facilitate incorporation of the antitumor drug, doxorubicin (DOX), up to a maximum loading of 80 µmol/g. In vitro cell uptake of nanocomposites was performed with DLD-1 colon cancer cells and PLC-PRF-5 liver cancer cells. Confocal microscopy photos illustrate that doxorubicin-loaded nanoparticles accumulate in both the cytoplasm and the cell nuclei. Cell survival efficiency with maghemite nanocomposites was determined via the MTT assay, and the cytotoxicity study proved that they exhibited significant toxicity against both types of cancer cells, although the improvement over free DOX treatment is more evident in the case of DLD-1 cancer cells when the most dilute drug and particle solutions are compared.

Histological and immunohistochemical assessment of liver biopsies in morbidly obese patients.

AIMS: To study liver lesions in morbidly obese patients who underwent liver biopsy at the time of bariatric surgery to define histological lesions, especially inflammatory infiltrate, diagnostic categories and the possible influence of gender in this respect. METHODS AND RESULTS: 110 biopsies (36 males-M- and 76 females -F-) were evaluated and categorised, according to the NAS (NAFLD -non alcoholic fatty liver disease- Activity Score) system and other criteria, as non-NAFLD (15.5%, F predominance), non-alcoholic steatohepatitis (NASH) (16.5%, M predominance), non-alcoholic hepatosteatosis (NAHS) (21%, F predominance) and, the most numerous group, NASH-borderline (NASH-BORD) (47%), with three subgroups, characterised by centrozonal lesions, portal area preferential involvement or affecting both areas. The predominant form of hepatocytesteatosis was mixed with a multivesicular component that was present in most cases with fibroinflammatory portal involvement. Nuclear glycogenosomes were found in greater number of biopsies in patients in the third and sixth decades. Portal inflammation was present in a large number of cases (M predominance); the application of immunohistochemical techniques (myeloperoxidase and CD68 antibodies) to evaluate lobular inflammation revealed "surgical hepatitis" in one third of the cases, and the presence of microgranulomas (CD68+) (M predominance), which were more abundant with increasing lesion severity. CONCLUSIONS: Portal inflammation and multivesicular hepatocytesteatosis are highly prevalent in morbidly obese patients. This study identifies a new subtype of NASH-BORD characterized by centrizonal and porto-periportal area involvement and the existence of liver biopsies without steatosis. CD68+ microgranulomas constitute an unequivocal marker of lobular inflammation in surgical biopsies and of lesion severity, which is gender-related.

Inhibition of poly (ADP-ribose) polymerase-1 enhances doxorubicin activity against liver cancer cells.

The purpose of this study was to investigate whether PARP-1 inhibition sensitizes human liver cancer cell lines to doxorubicin treatment. Both the addition of PARP-1 inhibitor (ANI) and depletion by means of stable siRNA significantly enhanced the growth inhibition induced by the DNA damage agents used. This effect was associated with an accumulation of unrepaired DNA, with a reduction in EGFR and Bcl-xL gene expression as well as with positive annexin-V staining. These results provide novel evidence of the direct role of PARP-1 in tumour chemoresistance in relation to its effects on the transcription of key genes involved in tumour survival.

[Laryngeal and hypopharyngeal liposarcoma. Report of a case and literature review]. / Liposarcoma de laringe e hipofaringe. Presentación de un caso y revisión de la literatura.

Liposarcoma is the second sarcoma of soft tissues more frequent in adult-age. Its localisation in head and neck is odd. The majority of liposarcoma, of this sitting, take its origin from soft tissues of the neck, being unusual primary liposarcoma from larynx and hypolarynx. Presentation of one case of liposarcoma of the larynx and hypopharynx and review of the opportune literature.

Criterios terapéuticos en la oftalmopatía asociada a enfermedad tiroidea. Análisis en la comunidad andaluza / Current therapy of thyroid-associated ophthalmology. Andalusian community survey

Antecedentes. El tratamiento más adecuado para la oftalmopatía asociada a la enfermedad de Graves moderada o grave no ha sido establecido. El objetivo de este estudio fue investigar las tendencias en el diagnóstico y tratamiento de la oftalmopatía tiroidea y el hipertiroidismo coexistente. Método. Se envió por correo un cuestionario sobre un caso típico y algunas variantes de oftalmopatía de Graves a 15 Unidades de Endocrinología del Servicio Andaluz de Salud entre octubre y noviembre de 2001. Se recibieron 42 respuestas de todas y cada una de las unidades. Los resultados se compararon con los obtenidos en un estudio europeo de Weetman y Wiersinga realizado en 1998.Resultados. Los corticoides por vía oral (73,6 por ciento) y en menor grado por vía intravenosa (14,3 por ciento) fueron el tratamiento más utilizado para la oftalmopatía en el caso índice, seguidos de radioterapia asociada a esteroides por vía oral o intravenosa el 28 por ciento y cirugía descompresiva el 12 por ciento. El empeoramiento de los signos oculares después de 8 semanas desplazó el uso de esteroides orales hacia radioterapia más esteroides por cualquier vía de administración (56 por ciento), cirugía descompresiva (40 por ciento) y esteroides intravenosos (33 por ciento). Para el tratamiento del hipertiroidismo coexistente, los fármacos antitiroideos fueron elegidos por un 93 por ciento de los encuestados. En las recaídas del hipertiroidismo, la tiroidectomía fue preferida por el 71 por ciento y el radioyodo se usó principalmente asociado a dosis profilácticas de esteroides orales. Solamente existieron diferencias, en comparación con el estudio europeo, en la utilización de otros inmunodepresores y de análogos de la somatostatina en la progresión de la enfermedad, y el régimen de antitiroideos (ajuste frente a bloqueoreemplazo) para el hipertiroidismo. Conclusiones. Los corticoides por vía oral son aún el tratamiento más extensamente utilizado en pacientes con oftalmopatía tiroidea; los antitiroideos son el tratamiento de elección en el hipertiroidismo coexistente y la tiroidectomía es preferida en las recurrencias (AU)

Soaking: the effect of osmotic shock on tetragonal lysozyme crystals.

Protein crystals crack when they are soaked in a solution with ionic strength sufficiently different from the environment in which they grew. It is demonstrated for the case of tetragonal lysozyme that the forces involved and the mechanisms that lead to the formation of cracks are different for hypertonic and hypotonic soaking. Tetragonal lysozyme crystals are very sensitive to hypotonic shocks and, after a certain waiting time, cracks always appear with a characteristic pattern perpendicular to the crystallographic c axis. Conversely, a hypertonic shock is better withstood: cracks do not display any deterministic pattern, are only visible at higher differences in ionic strength and after a certain time a phenomenon of crystal reconstruction occurs and the cracks vanish. At the lattice level, the unit-cell volume expands in hypotonic shock and shrinks under hypertonic conditions. However, the compression of the unit cell is anisotropic: the c axis is compressed to a minimum, beyond which it expands despite the unit-cell volume continuing to shrink. This behaviour is a direct consequence of the positive charge that the crystals bear and the existence of channels along the crystallographic c axis. Both features are responsible for the Gibbs-Donnan effect which limits the free exchange of ions and affects the movement of water inside the channels and bound to the protein.

The reversal of the inhibition on lipids synthesis by L-659,699 in arterial smooth muscle cells cultures.

The beta-lactone isolated from Fusarium sp. termed L-659,699 is a potent specific inhibitor of the enzyme 3-hydroxi-3-methylglutaril coenzyme A (HMG-CoA) synthase. In cultures of smooth muscle cells (SMC) isolated from aortic-arch of control (C-SMC) and 5% of cholesterol diet (Ch-SMC) treated chicks, the incorporation of (14C)-acetate to lipids (cholesterol, triacylglycerides and cholesterol ester) were greater in Ch-SMC cultures than in C-SMC and the presence of 0.05 microM L-659,699 for 2 h in the incubation medium decrease the synthesis of cholesterol however the triacylglycerides synthesis increase. The effect of inhibitor is stronger in young cultures (3-4 steps) than in the older ones (11-12 steps). In young C-SMC and Ch-SMC cultures the inhibition of cholesterol and triacylglycerides synthesis by L-659,699 was reversal.

Hipertiroidismo inducido por amiodarona / Amiodarone-induced hyperthyroidism

La amiodarona es un fármaco utilizado extensamente en la práctica cardiológica por sus excelentes propiedades antiarrítmicas. Produce alteraciones funcionales tiroideas por su alto contenido en yodo, contiene un 37 por ciento de yodo y es estructuralmente similar a las hormonas tiroideas. Produce inhibición hepática de la 5'desyodas.El hipertiroidismo secundario a amiodarona presenta una incidencia ente el 6-12 por ciento de los pacientes tratados, siendo en la infancia las cifras de incidencias similares. La determinación de niveles de T3, T4, TSH juegan un papel importante por el seguimiento y diagnóstico de las alteraciones tiroideas. Las opciones de tratamiento del hipertiroidismo inducido por amiodarona en niños incluyen: tionamidas, perclorato potásico y prednisona. Presentamos un caso de hipertiroidismo secundario a amiodarona en un varón de 10 años con síndrome de Marfan que presenta varios ingresos por crisis de taquicardia paroxística supraventricular y fibrilación auricular. Tras tratamiento con amiodarona presenta un cuadro clínico y analítico de hipertiroidismo con cifras de TSH muy disminuidas y de T4 libre aumentadas. La ecografía y gammagrafía tiroideas fueron normales. Se inicia tratamiento con tiamazol no cediendo el cuadro, que se normaliza tras la administración de prednisona. Actualmente se retira lentamente la prednisona (AU)

Alterations in 3-hydroxy-3-methylglutaryl-CoA reductase mRNA concentration in cultured chick aortic smooth muscle cells.

We observed and compared alterations in 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase at the transcriptional level in unsynchronized, three-passage cultures of smooth-muscle cells from the aorta of chicks fed on a control diet (C-SMC) and those of chicks fed on a similar diet plus cholesterol (Ch-SMC). Alterations in reductase mRNA concentrations in senescent cultures were much lower. We used a modification of the competitive (c) reverse transcription polymerase chain reaction method, using a Thermus thermophilus DNA polymerase (Tth pol) to quantify the very scarce species of HMG-CoA reductase mRNA in samples of cytoplasmic SMC mRNA. We cloned and sequenced a 199 bp cDNA fragment of chicken HMG-CoA reductase, which encoded a region of 66 amino acids belonging to the catalytic domain of the enzyme. HMG-CoA reductase mRNA concentrations from young C-SMC cultures rose 3.89-fold 4 h after the change of medium and returned to base levels between 8 to 12 h afterward. Concentrations in Ch-SMC cultures increased less (2.36-fold) 8 h after the change to fresh medium. Increases in reductase mRNA in senescent cultures of Ch-SMC and C-SMC measured under similar conditions were only 1.28- and 1.39-fold, respectively.

Changes in cultured arterial smooth muscle cells isolated from chicks upon cholesterol feeding.

We have developed cultures of smooth muscle cells (SMC) isolated from arterial hypercholesterolemic chicks (cholesterol-SMC). These cultures are suitable for the study at the molecular level of the changes in arterial SMC induced by a cholesterol diet. By using a strong dose of cholesterol (5%) for 10 d, we obtained very proliferative SMC which became foam cells after 30 d in culture. On the other hand, SMC cultures isolated from control-fed chicks had a lower growth rate than the SMC ones under the same culture conditions. DNA synthesis was fourfold greater in cholesterol-SMC than in control-SMC cultures. Intracellular cholesterol concentrations were the same in both cholesterol and control SMC during the first 14 d of culture but afterward increased in differing ways: after 20 d of culture the cholesterol-SMC increased their cholesterol content to double the control. We give here the results obtained from transmission electron microscopy, lipid analysis, proliferation studies, DNA, RNA and protein synthesis, and then discuss their implications.

Glia protect fetal midbrain dopamine neurons in culture from L-DOPA toxicity through multiple mechanisms.

Mesencephalic glia produce soluble factors that protect dopamine neurons from L-DOPA toxicity. The chemical composition of these soluble factors is unknown. We investigated the protective effect against L-DOPA neurotoxicity in midbrain dopamine neurons of fractions of different molecular size of glia conditioned medium and candidate neuroprotective agents produced by glia including neurotrophic factors and antioxidants. Protective effects were evaluated according to the number of tyrosine hydroxylase immunoreactive cells, high affinity dopamine uptake and levels of quinones. Both fractions of glia conditioned medium, smaller and larger than 10kD, protected against L-DOPA, but the fraction of smaller molecular size, that contains small free radical scanvenger molecules, was more effective than the fraction of larger molecular size, that contains large neurotrophic peptides. Among the neurotrophic factors GDNF and BDNF totally prevented L-DOPA neurotoxicity, while NGF and bFGF were less effective. However, only NGF significantly reduced the elevation of quinones induced by L-DOPA. Ascorbic acid, at the concentration found in glia conditioned medium, provided partial protective effect against L-DOPA toxicity. Glutathione, had neurotrophic effects on untreated midbrain dopamine neurons and prevented the effect of L-DOPA. In conclusion, the protective effect against L-DOPA neurotoxicity by glia conditioned medium is mediated by several compounds including neurotrophic factors and small antioxidants.

Glia conditioned medium protects fetal rat midbrain neurones in culture from L-DOPA toxicity.

L-DOPA kills dopamine neurones in culture but is the most effective drug for the treatment of Parkinson's disease, where it exhibits no clear toxicity. While glial cells surround and protect neurones in vivo, neurones are usually cultured in vitro in the absence of glia. We treated fetal midbrain rat neurones with L-DOPA, mesencephalic glia conditioned medium (CM) and L-DOPA + CM. L-DOPA reduced the number of tyrosine hydroxylase-positive (TH+) cells and [3H]DA uptake, and increased quinone levels. L-DOPA + CM restored [3H]DA uptake and quinone levels to normal, and increased the number of TH+ cells and terminals to 170% of control. CM greatly increased the number of TH+ cells and [3H]DA uptake. Mesencephalic glia therefore produced soluble factors which are neurotrophic for dopamine neurones, and which protect these neurones from the toxic effects of L-DOPA.

Changes induced by ovariectomy on the acute effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in a model of rat poor metabolizer of debrisoquine.

The female 'dark-adapted' rat, an animal model of poor metabolizer of debrisoquine, is more susceptible to neurotoxic effects of 1,2,3,6-tetrahydropyridine (MPTP) than other rat species (extensive metabolizers). Since ovariectomy improves the ability for the 4-hydroxylation of debrisoquine in female 'dark-adapted' rats, we studied the acute effects of MPTP (three doses of 10, 20, and 30 mg/kg s.c., respectively, at 12 hour intervals) in ovariectomized and laparotomized female 'dark-adapted' rats to test whether ovariectomy prevents MPTP-induced neurotoxicity. We measured regional brain monoamine levels. MPTP-induced depletion of dopamine (DA) and its metabolites was more prominent in the striatum. Ovariectomy, by itself, reduced dopamine and serotonin (5-HT) and their metabolites in striatum in both control and MPTP-treated animals. Factorial analysis of the effects of ovariectomy and MPTP treatment by two-way ANOVA revealed that both experimental procedures reduced DA and 5-HT neurotransmission in the striatum while the combined effect of both treatments did not produce any significant change. In spite of its induction of monoamine depletion in intact animals, ovariectomy partially prevented MPTP-induced depletion of monoamines in the surviving rats, suggesting that changes in metabolic rates of debrisoquine induced by ovariectomy produce resistance to MPTP in 'dark-adapted' rats.

[Salmonellosis and ulcerative colitis. A case report and review of the literature]. / Salmonelosis y colitis ulcerosa. Presentación de un caso y revisión de la literatura.

A propos of a case of intestinal infection due to Salmonella non typhi in a patient with ulcerative colitis, we have reviewed the disease and, therefore ran a search through the scientific literature in which we have found 16 cases of ulcerative colitis diagnosed in a clinical setting which began as an enterocolitis due to Salmonella. Bacteriological studies, clinical evolution and response to treatment, normally allow to differentiate between the two clinical entities. Colitis due to Salmonella and ulcerative colitis can coexist in the same patient. Duration of the clinical manifestations oven four weeks, previous history of diarrhea with blood, the sudden worsening of a mild prolonged diarrhea, non-response to specific antibiotic treatment and fast response to therapy with corticoids of a diarrhea positive isolation of Salmonella in stools, should make us immediately think in the coexistence of a subjacent ulcerative colitis. Treatment with steroids always must be associated with systemic antibiotics if Salmonella has been isolated just before or during their administration.