RESUMEN
We report that the apical dendrites of CA3 hippocampal pyramidal neurons are increased during labor and birth in the valproate model of autism but not in control animals. Using the iDISCO clearing method, we show that hippocampal, especially CA3 region, and neocortical volumes are increased and that the cerebral volume distribution shifts from normal to lognormal in valproate-treated animals. Maternal administration during labor and birth of the NKCC1 chloride transporter antagonist bumetanide, which reduces [Cl-]i levels and attenuates the severity of autism, abolished the neocortical and hippocampal volume changes and reduced the whole-brain volume in valproate-treated animals. These results suggest that the abolition of the oxytocin-mediated excitatory-to-inhibitory shift of GABA actions during labor and birth contributes to the pathogenesis of autism spectrum disorders by stimulating growth during a vulnerable period.
Asunto(s)
Trastorno del Espectro Autista/tratamiento farmacológico , Trastorno del Espectro Autista/fisiopatología , Bumetanida/uso terapéutico , Hipocampo/metabolismo , Parto/metabolismo , Células Piramidales/metabolismo , Inhibidores del Simportador de Cloruro Sódico y Cloruro Potásico/uso terapéutico , Animales , Animales Recién Nacidos , Trastorno del Espectro Autista/inducido químicamente , Dendritas/efectos de los fármacos , Dendritas/metabolismo , Modelos Animales de Enfermedad , Femenino , GABAérgicos/farmacología , Embarazo , Células Piramidales/efectos de los fármacos , Ratas , Ratas Wistar , Ácido Valproico/farmacologíaRESUMEN
White biotechnology is a term that is now often used to describe the implementation of biotechnology in the industrial sphere. Biocatalysts (enzymes and microorganisms) are the key tools of white biotechnology, which is considered to be one of the key technological drivers for the growing bioeconomy. Biocatalysts are already present in sectors such as the chemical and agro-food industries, and are used to manufacture products as diverse as antibiotics, paper pulp, bread or advanced polymers. This review proposes an original and global overview of highly complementary fields of biotechnology at both enzyme and microorganism level. A certain number of state of the art approaches that are now being used to improve the industrial fitness of biocatalysts particularly focused on the biorefinery sector are presented. The first part deals with the technologies that underpin the development of industrial biocatalysts, notably the discovery of new enzymes and enzyme improvement using directed evolution techniques. The second part describes the toolbox available by the cell engineer to shape the metabolism of microorganisms. And finally the last part focuses on the 'omic' technologies that are vital for understanding and guide microbial engineering toward more efficient microbial biocatalysts. Altogether, these techniques and strategies will undoubtedly help to achieve the challenging task of developing consolidated bioprocessing (i.e. CBP) readily available for industrial purpose.
Asunto(s)
Bacterias/enzimología , Biocatálisis , Biotecnología , Enzimas/metabolismo , Bacterias/química , Enzimas/química , Enzimas/genética , Humanos , IndustriasRESUMEN
We report here the in vivo production of type 2 fucosylated-N-acetyllactosamine oligosaccharides in Escherichia coli. Lacto-N-neofucopentaose Galbeta1-4GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3)Glc, lacto-N-neodifucohexaose Galbeta1-4(Fucalpha1-3)Glc-NAcbeta1-3Galbeta1-4(Fucalpha1-3)Glc, and lacto-N-neodifucooctaose Galbeta1-4GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3)GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3)Glc were produced from lactose added in the culture medium. Two of them carry the Lewis X human antigen. High cell density cultivation allowed obtaining several grams of fucosylated oligosaccharides per liter of culture. The fucosylation reaction was catalyzed by an alpha-1,3 fucosyltransferase of Helicobacter pylori overexpressed in E. coli with the genes lgtAB of N. meningitidis. The strain was genetically engineered in order to provide GDP-fucose to the system, by genomic inactivation of gene wcaJ involved in colanic acid synthesis and overexpression of RcsA, positive regulator of the colanic acid operon. To prevent fucosylation at the glucosyl residue, lactulose Galbeta1-4Fru was assayed in replacement of lactose. Lactulose-derived oligosaccharides carrying fucose were synthesized and characterized. Fucosylation of the fructosyl residue was observed, indicating a poor acceptor specificity of the fucosyltransferase of H. pylori.
Asunto(s)
Escherichia coli/genética , Fucosa/metabolismo , Fucosiltransferasas/metabolismo , Ingeniería Genética , Helicobacter pylori/enzimología , Oligosacáridos/metabolismo , Cromatografía en Gel , Cromatografía en Capa Delgada , Escherichia coli/citología , Fermentación , Fucosiltransferasas/genética , Helicobacter pylori/genética , Cinética , Lactosa/metabolismo , Lactulosa/metabolismo , Espectroscopía de Resonancia Magnética , Metilación , Oligosacáridos/química , Oligosacáridos/aislamiento & purificación , Espectrometría de Masa Bombardeada por Átomos VelocesRESUMEN
The relationship between ritonavir plasma concentration, efficacy, and tolerance was evaluated in 31 children with advanced HIV infection who were receiving a triple therapy with ritonavir as protease inhibitor. Median CD4+ lymphocyte count and median viral load before the initiation of ritonavir-containing combination therapy were 1320 cells/mL and 5 log10 copies/mL, respectively. Ritonavir was given at a dose ranging from 300 to 450 mg/m2 twice daily. The median follow-up of triple therapy was 19 months. Response was defined as a drop of viremia of more than 1 log. Plasma drug levels were determined twice during the observation period: after at least 4 weeks and after 3 months of combined treatment. Samples were collected before (residual) and 2 hours (T2) after drug intake. Cholesterol, triglycerides, alanine transaminase, aspartate transaminase, and gamma-glutamyl transpeptidase were assessed at the same time. The median values of ritonavir residual and T2 levels were 1.64 mg/L and 5.9 mg/L at observation 1 and 3.35 mg/L and 6.29 mg/L at observation 2, respectively. According to virologic response, median residual concentrations of ritonavir were 3.17, 2.52, and 1.04 mg/L for the complete, the partial, and the no-response groups. The authors observed a wide intersubject variability of ritonavir concentrations with an increase in residual levels between the two observation periods. Residual levels were correlated with virologic response whereas there was no direct association between T2 levels and long-term response. Patients with complete or partial response displayed statistically significantly higher residual concentrations than the no-response group. No correlation could be demonstrated between elevated plasma drug concentrations and abnormal cholesterol or triglycerides values. These results emphasize the importance of a sustained high ritonavir concentration to achieve optimal treatment efficacy. Furthermore, these results prove the clinical benefit of therapeutic drug monitoring and could potentially improve patient evaluation in terms of treatment efficacy, compliance, and viral resistance.
Asunto(s)
Infecciones por VIH/tratamiento farmacológico , Inhibidores de la Proteasa del VIH/sangre , Ritonavir/sangre , Adolescente , Niño , Preescolar , Colesterol/sangre , Citocromo P-450 CYP3A , Sistema Enzimático del Citocromo P-450/fisiología , Monitoreo de Drogas , Residuos de Medicamentos/análisis , Humanos , Lactante , Hígado/efectos de los fármacos , Oxigenasas de Función Mixta/fisiología , Estudios Retrospectivos , Ritonavir/efectos adversos , Ritonavir/uso terapéutico , Triglicéridos/sangreRESUMEN
Jejunal diverticula is rare and in most cases without any symptoms. They become clinically relevant when complications, such as diverticulitis, malabsorption caused by bacterial overgrowth, intestinal hemorrhage, or obstruction, occur. In this case report a case of perforated jejunal diverticulitis is presented and the problems in finding the correct diagnosis are discussed.
Asunto(s)
Abdomen Agudo/etiología , Diverticulitis/diagnóstico por imagen , Perforación Intestinal/diagnóstico por imagen , Enfermedades del Yeyuno/diagnóstico por imagen , Tomografía Computarizada por Rayos X , Abdomen Agudo/patología , Abdomen Agudo/cirugía , Anciano , Anciano de 80 o más Años , Diagnóstico Diferencial , Diverticulitis/patología , Diverticulitis/cirugía , Humanos , Perforación Intestinal/patología , Perforación Intestinal/cirugía , Enfermedades del Yeyuno/patología , Enfermedades del Yeyuno/cirugía , Yeyuno/diagnóstico por imagen , Yeyuno/patología , Yeyuno/cirugía , MasculinoRESUMEN
Sixty-seven pregnant bitches were given atropine sulphate (0.025 mg kg-1), prifinium bromide (0.1 ml kg-1) and metopimazine (0.5 mg kg-1) and 15 min later 2.5 micrograms cloprostenol kg-1 s.c., three times at 48 h intervals (day 1, day 3, day 5). After one treatment, 53 of the 67 bitches had aborted, and after a second treatment, 62 of the 67 bitches had aborted. In 18 bitches, progesteronemia kinetics were followed-up: the first injection of cloprostenol resulted in a significant (P < 0.01) fall in progesteronemia. In 12 of the 18 bitches that had aborted following the first protocol, this rapid fall in progesterone was noteworthy as it decreased progesterone concentration on average from 17.07 +/- 8.20 ng ml-1 on day 1 to 1.31 +/- 0.34 ng ml-1 on day 3. The premedication administered 15 min before the injection of prostaglandins, prevented the appearance of side effects in 39 of the 67 bitches (58.2%).