RESUMEN
In Huntington's disease (HD), the mutant huntingtin (mHtt) accumulates as toxic aggregates in the striatum tissue, with deleterious effects on motor-coordination and cognitive functions. Reducing the levels of mHtt is therefore a promising therapeutic strategy. We have previously reported that GSK-3 is a negative regulator of the autophagy/lysosome pathway, which is responsible for intracellular degradation, and is critically important for maintaining neuronal vitality. Thus, we hypothesized that inhibition of GSK-3 may trigger mHtt clearance thereby reducing mHtt cytotoxicity and improving HD symptoms. Here, we demonstrate that depletion or suppression of autophagy results in a massive accumulation of mHtt aggregates. Accordingly, mHtt aggregates were localized in lysosomes, but, mostly mislocalized from lysosomes in the absence of functional autophagy. Overexpression of GSK-3, particularly the α isozyme, increased the number of mHtt aggregates, while silencing GSK-3α/ß, or treatment with a selective GSK-3 inhibitor, L807mts, previously described by us, reduced the amounts of mHtt aggregates. This effect was mediated by increased autophagic and lysosomal activity. Treating R6/2 mouse model of HD with L807mts, reduced striatal mHtt aggregates and elevated autophagic and lysosomal markers. The L807mts treatment also reduced hyperglycemia and improved motor-coordination functions in these mice. In addition, L807mts restored the expression levels of Sirt1, a critical neuroprotective factor in the HD striatum, along with its targets BDNF, DRPP-32, and active Akt, all provide neurotrophic/pro-survival support and typically decline in the HD brain. Our results provide strong evidence for a role for GSK-3 in the regulation of mHtt dynamics, and demonstrate the benefits of GSK-3 inhibition in reducing mHtt toxicity, providing neuroprotective support, and improving HD symptoms.
Asunto(s)
Glucógeno Sintasa Quinasa 3/antagonistas & inhibidores , Glucógeno Sintasa Quinasa 3/metabolismo , Proteína Huntingtina/metabolismo , Enfermedad de Huntington/metabolismo , Enfermedad de Huntington/patología , Animales , Línea Celular Tumoral , Glucógeno Sintasa Quinasa 3/genética , Humanos , Proteína Huntingtina/genética , Enfermedad de Huntington/genética , Lisosomas/metabolismo , Masculino , Ratones , Ratones Endogámicos CBA , Ratones TransgénicosRESUMEN
CRISPR-Cas systems provide prokaryotes with sequence-specific immunity against viruses and plasmids based on DNA acquired from these invaders, known as spacers. Surprisingly, many archaea possess spacers that match chromosomal genes of related species, including those encoding core housekeeping genes. By sequencing genomes of environmental archaea isolated from a single site, we demonstrate that inter-species spacers are common. We show experimentally, by mating Haloferax volcanii and Haloferax mediterranei, that spacers are indeed acquired chromosome-wide, although a preference for integrated mobile elements and nearby regions of the chromosome exists. Inter-species mating induces increased spacer acquisition and may result in interactions between the acquisition machinery of the two species. Surprisingly, many of the spacers acquired following inter-species mating target self-replicons along with those originating from the mating partner, indicating that the acquisition machinery cannot distinguish self from non-self under these conditions. Engineering the chromosome of one species to be targeted by the other's CRISPR-Cas reduces gene exchange between them substantially. Thus, spacers acquired during inter-species mating could limit future gene transfer, resulting in a role for CRISPR-Cas systems in microbial speciation.
Asunto(s)
Sistemas CRISPR-Cas/genética , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas/genética , ADN Intergénico/genética , Transferencia de Gen Horizontal/genética , Haloferax mediterranei/genética , Haloferax volcanii/genética , Especiación Genética , Haloferax mediterranei/crecimiento & desarrollo , Haloferax volcanii/crecimiento & desarrolloRESUMEN
In patients with concomitant occurrence of maxillofacial and basilar skull fractures, open reduction and internal fixation is the treatment. It requires intermittent intra operative dental occlusion which precludes oral or nasal intubation. In such cases submental intubation (SMI) is a recognized technique in practice. We describe a modified technique for smooth exteriorization of the endotracheal tube (ETT) during SMI. As the SMI technique is unusual for the performer, emphasis is laid on the applied aspects to minimize probable complications during the procedure. With the modified technique we performed SMI uneventfully on five patients.
RESUMEN
Latency between an early neurological insult and development of spontaneous recurrent seizures suggests aberrant chronological reorganization in patients with mesial temporal sclerosis associated epilepsy. Kainate-induced status similarly results in delayed development of spontaneous recurrent seizures. Mossy fiber sprouting by the dentate granule cells is a well-characterized manifestation of such temporal structural reorganization in both patients and animal models. However, alterations in other components of hippocampal circuitry have not been evaluated. We present results from studies using precise anterograde and retrograde tract tracing methodologies to evaluate the reorganization of outflow of the CA3 pyramidal cells. Although septotemporal relationships of the normal CA3 outflow tract through the Schaffer collaterals are well known, their aberrant reorganization following kainate-induced spontaneous recurrent seizures is not known. We provide the first definitive evidence of widespread CA3 structural reorganization in the form of sprouting of CA3 axons to widespread areas throughout the hippocampus and entorhinal cortex. This includes an apparent increase in the density of projection to areas that normally receive CA3 outflow such as CA1 and subiculum as well as novel projections beyond the confines of the hippocampus to the pre and parasubiculum and medial and lateral entorhinal cortex. We provide the first evidence of novel CA3 Schaffer collateral projection to the entorhinal cortex. The sprouting of CA3 outflow to widespread regions of the hippocampus and the entorhinal cortex may provide insight into how the injured hippocampus propagates unconventional impulse excitation to cortical fields which have a critical role in providing excitatory inputs into the hippocampus possibly setting up reverberating excitatory circuits as well as widespread connections throughout the cortical mantle. Sprouting-related mechanisms may also explain the latency associated with development of spontaneous recurrent seizures, the hallmark of temporal lobe epilepsy.