The evolution of the huntingtin-associated protein 40 (HAP40) in conjunction with huntingtin

Background The huntingtin-associated protein 40 (HAP40) abundantly interacts with huntingtin (HTT), the protein that is altered in Huntington's disease (HD). Therefore, we analysed the evolution of HAP40 and its interaction with HTT. Results We found that in amniotes HAP40 is encoded by a single-exon gene, whereas in all other organisms it is expressed from multi-exon genes. HAP40 co-occurs with HTT in unikonts, including filastereans such as Capsaspora owczarzaki and the amoebozoan Dictyostelium discoideum, but both proteins are absent from fungi. Outside unikonts, a few species, such as the free-living amoeboflagellate Naegleria gruberi, contain putative HTT and HAP40 orthologs. Biochemically we show that the interaction between HTT and HAP40 extends to fish, and bioinformatic analyses provide evidence for evolutionary conservation of this interaction. The closest homologue of HAP40 in current protein databases is the family of soluble N-ethylmaleimide-sensitive factor attachment proteins (SNAPs). Conclusion Our results indicate that the transition from a multi-exon to a single-exon gene appears to have taken place by retroposition during the divergence of amphibians and amniotes, followed by the loss of the parental multi-exon gene. Furthermore, it appears that the two proteins probably originated at the root of eukaryotes. Conservation of the interaction between HAP40 and HTT and their likely coevolution strongly indicate functional importance of this interaction

Free Radicals, Salicylic Acid and Mycotoxins in Asparagus After Inoculation with Fusarium proliferatum and F. oxysporum

Electron paramagnetic resonance was used to monitor free radicals and paramagnetic species like Fe, Mn, Cu generation, stability and status in Asparagus officinalis infected by common pathogens Fusarium proliferatum and F. oxysporum. Occurrence of F. proliferatum and F. oxysporum, level of free radicals and other paramagnetic species, as well as salicylic acid and mycotoxins content in roots and stems of seedlings were estimated on the second and fourth week after inoculation. In the first term free and total salicylic acid contents were related to free radicals level in stem (P = 0.010 and P = 0.033, respectively). Concentration of Fe3+ ions in porphyrin complexes (g = 2.3, g = 2.9) was related to the species of pathogen. There was no significant difference between Mn2+ concentrations in stem samples; however, the level of free radicals in samples inoculated with F. proliferatum was significantly higher when compared to F. oxysporum

Meeting Abstract: A12

Huntingtin (HTT) is the protein that is altered in Huntington’s disease (HD). While HTT has been found to be essential for many cellular activities including transport of vesicles in the cell, cellular uptake of materials (endocytosis), cellular degradation of waste (autophagy), regulation of transcription, and even is important for embryonic development, an integrating understanding of HTT’s many biological functions at the molecular level is still missing. Although the HTT gene and the disease-causing mutation have been identified 25 years ago, very little data has been available on the structure of HTT. However, information on HTT’s structure would be very important for achieving an improved understand of HTT’s function in health and disease. Here we employed cryo-electron microscopy (cryo-EM) to determine the structure of full-length human HTT in a complex with another protein, HTT-associated protein 40 (HAP40). This interaction of HTT with HAP40 was instrumental in stabilizing HTT’s structure to a degree that the structure of HTT could be determined at detailed level at an overall resolution of 4 Å. HTT is largely α-helical and consists of three major domains. The N- and C-terminal domains contain multiple HEAT repeat elements that are arranged in a solenoid fashion. These domains are connected by a smaller bridge domain that contain different types of tandem repeats. HAP40 is also largely α-helical and has a tetratricopeptide repeat (TPR)-like organization. HAP40 binds in a cleft contacting the three HTT domains by hydrophobic and electrostatic interactions, thereby stabilizing HTT’s conformation. These data help in the interpretation of previous biochemical results and will pave the way for the generation and testing of new research hypotheses that ultimately will lead to an improved understanding of HTT’s diverse biological functions

Case Report Emergence in Elderly Patient Undergoing General Anesthesia with Xenon

Introduction. It is a consensus that the postoperative cognitive function is impaired in elderly patients after general anaesthesia, and such category patient takes more time to recover. Xenon is a noble gas with anesthetic properties mediated by antagonism of Nmethyl-D-aspartate receptors. With a minimum alveolar concentration of 0.63, xenon is intended for maintaining hypnosis with 30% oxygen. The fast recovery after xenon anaesthesia was hypothesized to be advantageous in this scenario. Case Presentation. We report the case of 99-year-old woman who underwent sigmoid colon carcinoma resection with colorectal anastomosis. We carried out the induction phase by propofol, oxygen, fentanil, and rocuronium bromide, and then we proceeded to a rapid sequence endotracheal intubation consequently. The patient was monitored by IBP, NIBP, ECG, cardiac frequency, respiratory rate, capnometry, TOF Guard, blood gas analysis, and BIS. For maintenance we administrated oxygen, remifentanil, rocuronium bromide, and xenon gas 60-65%. Shortly after the end of surgery the patients started an autonomous respiratory activity, and a high BIS level was also recorded. Decision was made by our team to proceed into the emergence phase. The residual neuromuscular block was antagonized by sugammadex, modified Aldrete score was implicated, and we got our patient fully awake without any cognitive dysfunction or delirium. Conclusion. The rapid emergence to full orientation in very elderly patient who had been anesthetized by xenon shows concordance to the high BIS values and the clinical signs of the depth of anesthesia