Developmental Expression of Kv Potassium Channels at the Axon Initial Segment of Cultured Hippocampal Neurons

Axonal outgrowth and the formation of the axon initial segment (AIS) are early events in the acquisition of neuronal polarity. The AIS is characterized by a high concentration of voltage-dependent sodium and potassium channels. However, the specific ion channel subunits present and their precise localization in this axonal subdomain vary both during development and among the types of neurons, probably determining their firing characteristics in response to stimulation. Here, we characterize the developmental expression of different subfamilies of voltage-gated potassium channels in the AISs of cultured mouse hippocampal neurons, including subunits Kv1.2, Kv2.2 and Kv7.2. In contrast to the early appearance of voltage-gated sodium channels and the Kv7.2 subunit at the AIS, Kv1.2 and Kv2.2 subunits were tethered at the AIS only after 10 days in vitro. Interestingly, we observed different patterns of Kv1.2 and Kv2.2 subunit expression, with each confined to distinct neuronal populations. The accumulation of Kv1.2 and Kv2.2 subunits at the AIS was dependent on ankyrin G tethering, it was not affected by disruption of the actin cytoskeleton and it was resistant to detergent extraction, as described previously for other AIS proteins. This distribution of potassium channels in the AIS further emphasizes the heterogeneity of this structure in different neuronal populations, as proposed previously, and suggests corresponding differences in action potential regulation.

Computational Study of the Fe(CN)(2)CO Cofactor and Its Binding to HypC Protein.

In the intricate maturation process of [NiFe]-hydrogenases, the Fe(CN)2CO cofactor is first assembled in a HypCD complex with iron coordinated by cysteines from both proteins and CO is added after ligation of cyanides. The small accessory protein HypC is known to play a role in delivering the cofactor needed for assembling the hydrogenase active site. However, the chemical nature of the Fe(CN)2CO moiety and the stability of the cofactor–HypC complex are open questions. In this work, we address geometries, properties, and the nature of bonding of all chemical species involved in formation and binding of the cofactor by means of quantum calculations. We also study the influence of environmental effects and binding to cysteines on vibrational frequencies of stretching modes of CO and CN used to detect the presence of Fe(CN)2CO. Carbon monoxide is found to be much more sensitive to sulfur binding and the polarity of the medium than cyanides. The stability of the HypC–cofactor complex is analyzed by means of molecular dynamics simulation of cofactor-free and cofactor-bound forms of HypC. The results show that HypC is stable enough to carry the cofactor, but since its binding cysteine is located at the N-terminal unstructured tail, it presents large motions in solution, which suggests the need for a guiding interaction to achieve delivery of the cofactor.

The Spanish Travel Subjective Lexicon (STSL).

This paper presents a proposal for a recognition model for the appraisal value of sentences. It is based on splitting the text into independent sentences (full stops) and then analysing the appraisal elements contained in each sentence according to the previous value in the appraisal lexicon. In this lexicon, positive words are assigned a positive coefficient (+1) and negative words a negative coefficient (-1). We take into account word such as ?too?, ?little? (when it is not ?a bit?), ?less?, and ?nothing? than can modify the polarity degree of lexical unit when appear in the nearby environment. If any of these elements are present, then the previous coefficient will be multiplied by (-1), that is, they will change their sign. Our results show a nearly theoretical effectiveness of 90%, despite not achieving the recognition (or misrecognition) of implicit elements. These elements represent approximately 4% of the total of sentences analysed for appraisal and include the errors in the recognition of coordinated sentences. On the one hand, we found that 3.6 % of the sentences could not be recognized because they use different connectors than those included in the model; on the other hand, we found that in 8.6% of the sentences despite using some of the described connectors could not be applied the rules we have developed. The percentage relative to the whole group of appraisal sentences in the corpus was approximately of 5%.