Activation and closed-state inactivation mechanisms of the human voltage-gated KV4 channel complexes

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An inactivation stabilizer of the Na+ channel acts as an opportunistic pore blocker modulated by external Na+

The Na+ channel is the primary target of anticonvulsants carbamazepine, phenytoin, and lamotrigine. These drugs modify Na+ channel gating as they have much higher binding affinity to the inactivated state than to the resting state of the channel. It has been proposed that these drugs bind to the Na+ channel pore with a common diphenyl structural motif. Diclofenac is a widely prescribed anti-inflammatory agent that has a similar diphenyl motif in its structure. In this study, we found that diclofenac modifies Na+ channel gating in a way similar to the foregoing anticonvulsants. The dissociation constants of diclofenac binding to the resting, activated, and inactivated Na+ channels are approximately 880 microM, approximately 88 microM, and approximately 7 microM, respectively. The changing affinity well depicts the gradual shaping of a use-dependent receptor along the gating process. Most interestingly, diclofenac does not show the pore-blocking effect of carbamazepine on the Na+ channel when the external solution contains 150 mM Na+, but is turned into an effective Na+ channel pore blocker if the extracellular solution contains no Na+. In contrast, internal Na+ has only negligible effect on the functional consequences of diclofenac binding. Diclofenac thus acts as an "opportunistic" pore blocker modulated by external but not internal Na+, indicating that the diclofenac binding site is located at the junction of a widened part and an acutely narrowed part of the ion conduction pathway, and faces the extracellular rather than the intracellular solution. The diclofenac binding site thus is most likely located at the external pore mouth, and undergoes delicate conformational changes modulated by external Na+ along the gating process of the Na+ channel.     

The interactions of phenytoin and its binding site in DI-S6 segment of Na+ channel voltage-gated peptide by NMR spectroscopy and molecular modeling study.

Nuclear magnetic resonance (NMR) spectra of a model peptide (BL-DIS6), in the presence of anticonvulsant diphenyl drug, phenytoin (DPH), were measured to obtain the interactions between the selected drug and the model peptide. BL-DIS6's sequence corresponds to the S6 segment in domain I of rat brain type IIA Na+-channel. NMR studies have demonstrated that the magnitude of the chemical shifts of amide- and alpha-protons can be used as a measurement of the complex stability and binding site of the peptide. Our NMR results propose a 3(10)-helical structure for BL-DIS6, and suggest a binding cavity for DPH that involves the hydrophobic particles of residues Ans-7, Leu-8, Val-11, and Val-12. Furthermore, molecular modeling was performed to provide a possible complex conformation that the phenyl portion of DPH is accommodated in the proximity of the C-terminal residues Ala-11 and Val-12, and simultaneously the heterocyclic amine ring of DPH is perching at the residue Asn-7 periphery and stabilizing the phenyl portion deep insertion into the peptide.     

Prediction of radiotherapy response in cervix cancer by Raman spectroscopy: a pilot study

Radiotherapy is the choice of treatment for locally advanced stages of the cervical cancers, one of the leading female cancers. Because of intrinsic factors, tumors of same clinical stage and histological type often exhibit differential radioresponse. Radiotherapy regimen, from first fraction of treatment to clinical evaluation of response, spans more than 4 months. Clinical assessment by degree of tumor shrinkage is the only routinely practiced method to evaluate the tumor response. Hence, a need is created for development new methodologies that can predict the tumor response to radiotherapy at an early stage of the treatment which can lead to tailor-made protocols. To explore the feasibility of prediction of tumor radioresponse, Raman spectra of cervix cancer tissues that were collected before (malignant) and 24 h after patient was treated with 2nd fraction of radiotherapy (RT) were recorded. Data were analyzed by Principal Components Analysis (PCA) and results were correlated with clinical evaluation of radioresponse. Mean Raman spectra of RT tissues corresponding to different levels of tumor response, complete, partial, and no response, showed minute but significant variations. The unsupervised PCA of malignant tissues failed to provide any classification whereas RT spectra gave clear classification between responding (complete and partial response) and nonresponding conditions as well as a tendency of separation among responding conditions. These results were corroborated by supervised classification, by means of discrimination parameters: Mahalanobis distance and spectral residuals. Thus, findings of the study suggest the feasibility of Raman spectroscopic prediction of tumor radioresponse in cervical cancers.     

Cortical response induced by task-oriented training of the upper limb in subacute stroke patients as assessed by functional near-infrared spectroscopy

Despite the popularity of task-oriented training for stroke, the cortical reorganization associated with this type of therapy remains to be fully elucidated due to the lack of dynamic assessment tools. A good tolerance for motion artifacts makes functional near-infrared spectroscopy (fNIRS) suitable for investigating task-induced cortical responses in stroke patients. Here, patients were randomly assigned to receive task oriented (n = 25) or cyclic rotary training (n = 25) with simultaneous cortical activation and effective connectivity network analysis between prefrontal and motor cortices (PFC/MC). Compared with cyclic rotary training, task-oriented training induced significantly increased activation in both hemispheres and enhanced influence of PFC on MC. In addition, significantly decreased activation lateralization and increased betweenness centrality of the contralesional MC suggested widespread involvement of the contralesional hemisphere during task-oriented training. This study verifies the feasibility of fNIRS combined with motor paradigms for assessing neural responses associated with stroke rehabilitation in real time.     

Biochemical characterization of breast tumors by in vivo and in vitro magnetic resonance spectroscopy (MRS)

Magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS) have evolved as sensitive tools for anatomic and metabolic evaluation of breast cancer. In vivo MRS studies have documented the presence of choline containing compounds (tCho) as a reliable biochemical marker of malignancy and also useful for monitoring the tumor response to therapy. Recent studies on the absolute quantification of tCho are expected to provide cut-off values for discrimination of various breast pathologies. Addition of MRS investigation was also reported to increase the specificity of MRI. Further, ex vivo and in vitro MRS studies of intact tissues and tissue extracts provided several metabolites that were not be detected in vivo and provided insight into underlying biochemistry of the disease processes. In this review, we present briefly the role of various ¹H MRS methods used in breast cancer research and their potential in relation to diagnosis, monitoring of therapeutic response and metabolism.     

Quantification of glucosinolates in leaves of leaf rape (Brassica napus ssp. pabularia) by near-infrared spectroscopy

The potential of near-infrared spectroscopy (NIRS) for screening the total glucosinolate (t-GSL) content, and also, the aliphatic glucosinolates gluconapin (GNA), glucobrassicanapin (GBN), progoitrin (PRO), glucoalyssin (GAL), and the indole glucosinolate glucobrassicin (GBS) in the leaf rape (Brassica napus L. ssp. pabularia DC), was assessed. This crop is grown for edible leaves for both fodder and human consumption. In Galicia (northwestern Spain) it is highly appreciated for human nutrition and have the common name of "nabicol". A collection of 36 local populations of nabicol was analysed by NIRS for glucosinolate composition. The reference values for glucosinolates, as they were obtained by high performance liquid chromatography on the leaf samples, were regressed against different spectral transformations by modified partial least-squares (MPLS) regression. The coefficients of determination in cross-validation (r2) shown by the equations for t-GSL, GNA, GBN, PRO, GAL and GBS were, respectively, 0.88, 0.73, 0.81, 0.78, 0.37 and 0.41. The standard deviation to standard error of cross-validation ratio, were for these constituents, as follows: t-GSL, 2.96; GNA, 1.94; GBN, 2.31; PRO, 2.11; GAL, 1.27, and GBS, 1.29. These results show that the equations developed for total glucosinolates, as well as those for gluconapin, glucobrassicanapin and progoitrin, can be used for screening these compounds in the leaves of this species. In addition, the glucoalyssin and glucobrassicin equations obtained, can be used to identify those samples with low and high contents. From the study of the MPLS loadings of the first three terms of the different equations, it can be concluded that some major cell components as protein and cellulose, highly participated in modelling the equations for glucosinolates.