Clinical examples of 3D dose distribution reconstruction,based on the actual MLC leaves movement,for dynamic treatment techniques

AimTo present practical examples of our new algorithm for reconstruction of 3D dose distribution, based on the actual MLC leaf movement.BackgroundDynaLog and RTplan files were used by DDcon software to prepare a new RTplan file for dose distribution reconstruction.Materials and methodsFour different clinically relevant scenarios were used to assess the feasibility of the proposed new approach: (1) Reconstruction of whole treatment sessions for prostate cancer; (2) Reconstruction of IMRT verification treatment plan; (3) Dose reconstruction in breast cancer; (4) Reconstruction of interrupted arc and complementary plan for an interrupted VMAT treatment session of prostate cancer. The applied reconstruction method was validated by comparing reconstructed and measured fluence maps. For all statistical analysis, the U Mann–Whitney test was used.ResultsIn the first two and the fourth cases, there were no statistically significant differences between the planned and reconstructed dose distribution (p = 0.910, p = 0.975, p = 0.893, respectively). In the third case the differences were statistically significant (p = 0.015). Treatment plan had to be reconstructed.ConclusionDeveloped dose distribution reconstruction algorithm presents a very useful QA tool. It provides means for 3D dose distribution verification in patient volume and allows to evaluate the influence of actual MLC leaf motion on the dose distribution.     

Cheek tooth erosion in male babirusa (genus Babyrousa)

The wear on the occlusal surfaces of male babirusa cheek teeth was evaluated in 53 skulls of Babyrousa babyrussa from Buru and the Sula Islands and 87 skulls of B. celebensis from Sulawesi, Indonesia. Based on the comparative lengths of their continually growing maxillary canine teeth, the skulls were divided into five ‘age categories’ (A–E). Numerical and symbolic codes representing tooth wear were applied to each pillar (cusp region) of the mandibular and maxillary permanent third and fourth premolar teeth, and the first, second and third permanent molar teeth. There was no significant difference between the tooth wear patters of skulls in groups A and B, or in groups C and D, and so these were amalgamated. There was close correspondence in wear patterns between each side of the mouth in both species and in each age group. The wear patterns of the mandibular and maxillary teeth, although not identical, were very similar, as were the wear patterns of both species. In group A + B for both species tooth wear was relatively slight, with the M1 teeth experiencing most relative wear. There was almost no wear of the M3 teeth. In group C + D substantial wear of upper and lower M1 was evident. In group E more widespread wear of the cheek teeth was seen, with increased severity of M1 tooth wear, yet there was comparatively much less M2 and M3 tooth wear. The pattern of cheek tooth wear of the Babyrousa spp. was different from that shown by Sus scrofa. Differences in diet selection and processing were highlighted as potential contributing factors. The pattern of cheek tooth wear in male babirusa was not adequate for use to monitor their age.     

Growing-season precipitation since 1872 in the coastal area of subtropical southeast China reconstructed from tree rings and its relationship with the East Asian summer monsoon system

Tree rings from temperate zones of the world have provided abundant palaeo- ecological and paleo-hydroclimatic information. However, tree rings from subtropical to tropical regions remain relatively scarce, which greatly limit our fully understanding about the climate change issues. In the present work, tree-ring-width (TRW) measurements of Masson pine from Fujian province, the coastal area of subtropical southeast China were successfully crossdated and a TRW STD chronology was developed from 1854 to 2012. Significantly positive correlation was identified between the tree rings and April–November total precipitation (r = 0.71, p < 0.01). The reconstructed April–November precipitation exhibited two comparatively wet (1876–1886 and 1957–1962) and one comparatively dry (1986–2004) periods. An evident drying trend since 1959 was seen and it was mitigated after 1993. Most of the extreme low-precipitation years in the reconstruction were supported by the historical records. As revealed by the spatial correlation patterns, our precipitation reconstruction was also consistent with other hydroclimatic records along the coastal areas of southeast China, proving its ability to capture the large-scale hydrological signal in southeast China (mainly refers to the south of the middle-lower reaches of Yangtze River). The reconstructed precipitation showed significant correlation with the East Asian summer Monsoon (EASM) index. Moreover, it also indicated simultaneous variation with the monsoon precipitation in North China on a decadal scale, implying that growing season precipitation variations in both regions were influenced by the EASM strength. This work highlights the potential of using tree-ring width to reconstruct precipitation in subtropical southeast China, while the relevant issues about precipitation variation in this region is far from resolved.     

Maxillary tooth growth in the adult male babirusa (genus Babyrousa)

The growth of the maxillary canine teeth of the babirusa (genus Babyrousa) was studied on a sample of 149 adult male babirusa skulls from twenty-two international museum and private collections. Skulls from Buru, Sulawesi and the Togian Islands were represented. The continuous growth process was summarised into five stages (‘A’–‘E’) according to the position of the tip of the tooth over the bones of the skull. The supracutaneous portion of the tooth grew in a curve-linear fashion dorso-caudally, and was orientated such that the tips grew towards the midline of the cranium. The teeth of Sulawesi and Togian babirusa grew more dorsally over the nasal and frontal bones. Measurements were made on a subset of 45 teeth from Sulawesi babirusa skulls. The subcutaneous portion of the maxillary canine tooth (n = 22) increased in size from 37.3 mm (95% CI: 29.9–44.4 mm) in growth stage ‘B’ to 54.3 mm (49.4–59.2 mm) in growth stage ‘E’ as the erupted portion of the tooth (n = 19) lengthened from 81.3 mm (43.8–118.9 mm) in growth stage ‘A’ to 215.3 mm (177.8–252.9 mm) in growth stage ‘E’. The apical end of the tooth was open and thin-walled. The lumen of the tooth was filled with a cone of well vascularised dental pulp that extended almost to the tip of the tooth. The angle of curvature of the tooth within the alveolus (n = 22) decreased from 19.8 (17.3–22.3) degrees in growth stage ‘B’ to 7.4 (5.7–9.0) degrees in growth stage ‘E’. The corresponding supracutaneous angle of curvature (n = 25) reduced from 36.8 (33.6–40.1) degrees in growth stage ‘A’ to 10.7 (8.6–12.8) degrees in growth stage ‘E’.     

Urban–rural disparities of child health and nutritional status in China from 1989 to 2006

This paper analyzes urban–rural disparities of China's child health and nutritional status using the China Health and Nutrition Survey data from 1989 to 2006. We investigate degrees of health and nutritional disparities between urban and rural children in China as well as how such disparities have changed during the period 1989–2006. The results show that on average urban children have 0.29 higher height-for-age z-scores and 0.19 greater weight-for-age z-scores than rural children. Urban children are approximately 40% less likely to be stunted (OR = 0.62; p < 0.01) or underweight (OR = 0.62; p < 0.05) during the period 1989–2006. We also find that the urban–rural health and nutritional disparities have been declining significantly from 1989 to 2006. Both urban and rural children have increased consumption of high protein and fat foods from 1989 to 2006, but the urban–rural difference decreased over time. Moreover, the urban–rural gap in child preventive health care access was also reduced during this period.     

Pseudomonas silesiensis sp. nov. strain A3T isolated from a biological pesticide sewage treatment plant and analysis of the complete genome sequence

Microorganisms classified in to the Pseudomonas genus are a ubiquitous bacteria inhabiting variety of environmental niches and have been isolated from soil, sediment, water and different parts of higher organisms (plants and animals). Members of this genus are known for their metabolic versatility and are able to utilize different chemical compounds as a source of carbon, nitrogen or phosphorus, which makes them an interesting microorganism for bioremediation or bio-transformation. Moreover, Pseudomonas sp. has been described as a microorganism that can easily adapt to new environmental conditions due to its resistance to the presence of high concentrations of heavy metals or chemical pollution. Here we present the isolation and analysis of Pseudomonas silesiensis sp. nov. strain A3^T isolated from peaty soil used in a biological wastewater treatment plant exploited by a pesticide packaging company. Phylogenetic MLSA analysis of 4 housekeeping genes (16S rRNA, gyrB, rpoD and rpoB), complete genome sequence comparison (ANIb, Tetranucleotide identity, digital DDH), FAME analysis, and other biochemical tests indicate the A3^T strain (type strain PCM 2856^T = DSM 103370^T) differs significantly from the closest relative species and therefore represents a new species within the Pseudomonas genus. Moreover, bioinformatic analysis of the complete sequenced genome showed that it consists of 6,823,539 bp with a 59.58 mol% G + C content and does not contain any additional plasmids. Genome annotation predicted the presence of 6066 genes, of which 5875 are coding proteins and 96 are RNA genes.     

Evaluation of chemicals requiring metabolic activation in the EpiDerm™ 3D human reconstructed skin micronucleus (RSMN) assay

The in vitro human reconstructed skin micronucleus (RSMN) assay in EpiDerm? is a promising new assay for evaluating genotoxicity of dermally applied chemicals. A global pre-validation project sponsored by the European Cosmetics Association (Cosmetics Europe - formerly known as COLIPA), and the European Center for Validation of Alternative Methods (ECVAM), is underway. Results to date demonstrate international inter-laboratory and inter-experimental reproducibility of the assay for chemicals that do not require metabolism [Aardema et al., Mutat. Res. 701 (2010) 123–131]. We have expanded these studies to investigate chemicals that do require metabolic activation: 4-nitroquinoline-N-oxide (4NQO), cyclophosphamide (CP), dimethylbenzanthracene (DMBA), dimethylnitrosamine (DMN), dibenzanthracene (DBA) and benzo(a)pyrene (BaP). In this study, the standard protocol of two applications over 48 h was compared with an extended protocol involving three applications over 72 h. Extending the treatment period to 72 h changed the result significantly only for 4NQO, which was negative in the standard 48 h dosing regimen, but positive with the 72 h treatment. DMBA and CP were positive in the standard 48 h assay (CP induced a more reproducible response with the 72 h treatment) and BaP gave mixed results; DBA and DMN were negative in both the 48 h and the 72 h dosing regimens. While further work with chemicals that require metabolism is needed, it appears that the RMSN assay detects some chemicals that require metabolic activation (4 out of 6 chemicals were positive in one or both protocols). At this point in time, for general testing, the use of a longer treatment period in situations where the standard 48 h treatment is negative or questionable is recommended.     

In-depth exploration of Hevea brasiliensis latex proteome and “hidden allergens” via combinatorial peptide ligand libraries

The proteome of Hevea brasiliensis latex has been explored in depth via combinatorial peptide ligand libraries. A total of 300 unique gene products have been identified in this latex, whose proteome has been largely unknown up to the present. In search for unknown allergens, control latex and eluates from the ligand libraries have been fractionated by two-dimensional mapping, blotted and confronted with sera of 18 patients. In addition to the already known and named Hevea major allergens, we have unambiguously detected several others like, for instance: heat shock protein (81 kDa), proteasome subunit (30 kDa), protease inhibitor (8 kDa), hevamine A (43 kDa) and glyceraldehyde-3-phosphate dehydrogenase (37 kDa). Gene Ontology analysis of analyzed fractions has shown that major functions are substantially unchanged after sample treatment, while novel biological functions appeared that were undetectable in the crude sample.     

Synthesis and biological evaluation of some new 1,4-dihydropyridines containing different ester substitute and diethyl carbamoyl group as anti-tubercular agents

Tuberculosis is a leading infectious cause of death worldwide. Because of the concern of the resistance to most of the commonly used drugs displayed by the considered mycobacteria, most efforts have been done to introduce new anti-tubercular agents. Recent studies showed that 1,4-dihydropyridine-3,5-dicarbamoyl derivatives with lipophilic groups have significant anti-tubercular activity. In this study, we synthesized new derivatives of 1,4-dihydropyridines in which different alkyl and aryl esters and diethyl carbamoyl are substituted in C-3 and C-5 of the DHP ring. In addition nitroimidazole ring is substitutes at C-4 position. These asymmetric analogues were synthesized by a modified Hantzsh reaction using procedure reported by Meyer. The in vitro anti-tubercular activity of compounds against Mycobacterium tuberculosis was evaluated. The results indicate that the compounds containing aromatic esters are more potent than alkyl ones. The most potent aromatic compound (R = 3-phenylpropyl) exhibits comparable anti-tubercular activity (MIC = 1 μmol/ml) with reference compound isoniazide (INH) (MIC = 1 μmol/ml). Conformational analysis, SAR studies of these compounds showed that increasing in lipophilicity and rotable bonds of these compounds resulted in increasing anti-tubercular activity.     

Climatic information recorded in stable carbon isotopes in tree rings of Cryptomeria fortunei,Tianmu Mountain,China

The Cryptomeria fortunei (CF) tree-ring δ¹³C_p series, which was collected from the West Tianmu Mountain forestland (30°20′ N, 119°26′ E), located in the north-west of Zhejiang Province, China, belonging to the northern margin of the mid-subtropical region of Eastern China, were determined based on cross-dated tree-ring age. There was a significant decline in the δ¹³C_p series occurring from 1685 to 1985, more especially from 1835 to 1985 in response to increasing atmospheric CO₂ concentrations and decreasing atmospheric δ¹³C_a. To reduce the noise and enhance the climatic signals, we compared the polynomial function with the correction method developed by McCarroll and Loader (2004) to remove the low-frequency variation in the raw tree ring δ¹³C_p series (defined as the δ¹³C_poly series, δ¹³C_cor series, respectively), and found the most suited correction method was the correction method developed by McCarroll and Loader (2004) in our study area. High-frequency correlation analysis between the δ¹³C_cor series and many meteorological parameters recorded by Xian Rending weather station revealed that the current August–September mean maximum temperature and previous year mean minimum and mean maximum temperature (P < 0.005) most strongly influenced tree ring δ¹³C_p discrimination from 1956 to 1985, and the strongest temperature signal captured was the current August–September mean maximum temperature (r = 0.54, P < 0.005). Mainly on this basis, the varied history of current August–September mean maximum temperatures in the West Tianmu Mountain area were reconstructed from 1685 to 1985. The reconstructed maximum temperatures revealed a slight warming trend and showed close correlation with the climatic fluctuations of the Little Ice Age cold period before 1900 as well as the 20th century warm period after 1900. It also better corresponded with some climate events recorded in historical records. Spectrum analysis showed that in the reconstructed series there was quasi-periodicity of 66.7 yr, 21.1 yr, 3.2 yr, 2.3 yr and 2.0 yr. These cycles coincided with the “torque effect” variation of planets and the geocentric convergence, and changes in solar activity and irradiance, as well as the “Quasi-biennial oscillation” (QBO). This indicated that the δ¹³C_p chronology of tree rings in West Tianmu Mountain showed a good record of the sun's activities, the change in the sun radiation and ENSO events.     

June–September temperature reconstruction in the Northern Caucasus based on blue intensity data

Although long-lived trees grow in the Northern Caucasus, no single tree-ring chronology has been reported thus far from this area in the International Tree-Ring Data Base (ITRDB), neither has one been published in international journals. Extensive tree-ring studies were conducted over the last decade, and a tree-ring network was developed for the investigated area. The data on the minimum blue intensity based on 33 series of pine (Pinus sylvestris L.) and fir (Abies nordmanniana (Steven) Spach) is presented in this study. The minimum blue intensity (BI) chronology covers the period 1596–2011 with EPS value ≥0.85. The BI chronology strongly correlates with the mean June-September temperature (R = 0.74; p < 0.05) from the weather station “Kluhorskij Pereval” (1951–2011). Mean June-September temperature anomalies were reconstructed using the rescaling method. Based on the reconstruction provided in this study the twentieth century is characterized by highly increased June-September temperature. According to this study, the minimum blue intensity approach demonstrates a great potential for paleoclimatic research in the Caucasus. Vast spatial coverage of the new BI-based reconstruction based on data from only two locations in the Northern Caucasus provides prospects for reconstruction of temperature variations for a great region in the Middle East and Northern Africa.     

Preliminary assessment of somatic cell nuclear transfer in the dromedary (Camelus dromedarius)

Somatic cloning may enable the maintenance/expansion of the population of camels with the highest potential for milk production or the best racing performances. However, there have been no reports of embryonic or somatic nuclear transfer in camels. The aim of this study was to produce dromedary embryos by nuclear transfer using in vitro matured oocytes and two somatic cells from two sources (adult fibroblasts or granulosa cells). A total of 58 adult females were superstimulated by a single dose of eCG (3500 IU). Ten days later, their ovaries were collected postmortem. Cumulus–oocytes-complexes (COCs) were aspirated from stimulated follicles and were matured in vitro for 30 h. Fibroblasts (from live adult male) and granulosa cells (from slaughtered adult females) were used as donor karyoplasts and injected into mature enucleated dromedary oocytes.The cleavage rate was significantly higher (P < 0.05) for embryos reconstructed with fibroblasts (59%) versus those with granulosa cells (45%). However, there was no difference between the two groups in the proportion of cloned embryos reaching the blastocyst stage (fibroblasts: 14% vs. granulosa cells: 15%) or those that hatched (fibroblasts: 10% vs. granulosa cells: 12%). The viability of reconstructed dromedary embryos from the two sources of donor cells (fibroblasts; n = 5 vs. granulosa cells; n = 7) was examined by transferring them to synchronized recipients. Two females (fibroblasts: 1/5; 20%, granulosa cells: 1/7; 14%) were confirmed pregnant by ultrasonography at 15 and 25 days following transfer. Later, the pregnancies were followed by pregnancy empirical-symptoms. These two pregnancies were lost between 25 and 60 days following transfer, respectively.In conclusion, the present study shows for the first time that the development of dromedary NT embryos derived from either adult fibroblasts or granulosa cells can occur in vitro and the transfer of these cloned embryos to recipients can result in pregnancies.     

High allozymic diversity in natural populations of Mycoheterotrophic Orchid Gastrodia elata,an endangered medicinal plant in China

Gastrodia elata Blume, a mycoheterotrophic orchid native to the Far East, is an endangered medicinal plant in China. Genetic variation among 19 natural populations of G. elata was examined in central China by using allozyme polymorphism (16 loci in six enzymatic systems). The species exhibited high level of genetic diversity (P = 56.3%, A = 2.2 and H_E = 0.221), which was mainly attributed to its perennial habit and mixed reproduction system (both sexual and asexual). Evident genetic differentiation in G. elata natural populations was suggested by F_ST = 0.241. AMOVA analysis showed 31.3% of the total molecular variation was attributed to inter-population differentiation. Obvious genetic structure and genetic depauperation of some populations indicate forest fragmentation and over-collection have affected genetic variation of G. elata. A conservation strategy, which is conserving populations with great genetic distinction or high level of genetic variation from four management units, is recommended.     

Clinical characteristics of tumors derived from colorectal cancer patients who harbor the Tumor Necrosis Factor α-1031T/T and NOD2 3020insC polymorphism

Background: Genetic predispositions to disease have focused on highly penetrant causative changes in tumor suppressor genes or genes associated with DNA mismatch repair. New investigations are revealing new genetic associations with disease that are more subtle in their association with disease and require characterization. Methods: In this report we have examined the tumor characteristics in a group of patients who have been shown to harbor two polymorphisms in two genes that are associated with the immune system NOD2 and TNFα. Results: Colorectal cancers from patients with NOD2 3020insC and TNFα-1031T/T constitutional changes are mostly right-sided disease (OR = 2.21, p = 0.03) with a tendency to higher stages (OR = 2.41, p = 0.06), increased number of associated polyps (OR = 1.77, p = 0.16) and later age of average age of disease onset (p = 0.039). Conclusion: The results reveal that there appear to be specific characteristics associated with the tumors that may aid in determining management strategies to reduce the risk of disease.     

Molecular phylogeny of black fungus gnats (Diptera: Sciaroidea: Sciaridae) and the evolution of larval habitats

The phylogeny of the family Sciaridae is reconstructed, based on maximum likelihood, maximum parsimony, and Bayesian analyses of 4809 bp from two mitochondrial (COI and 16S) and two nuclear (18S and 28S) genes for 100 taxa including the outgroup taxa. According to the present phylogenetic analyses, Sciaridae comprise three subfamilies and two genus groups: Sciarinae, Chaetosciara group, Cratyninae, and Pseudolycoriella group + Megalosphyinae. Our molecular results are largely congruent with one of the former hypotheses based on morphological data with respect to the monophyly of genera and subfamilies (Sciarinae, Megalosphyinae, and part of postulated “new subfamily”); however, the subfamily Cratyninae is shown to be polyphyletic, and the genera Bradysia, Corynoptera, Leptosciarella, Lycoriella, and Phytosciara are also recognized as non-monophyletic groups. While the ancestral larval habitat state of the family Sciaridae, based on Bayesian inference, is dead plant material (plant litter + rotten wood), the common ancestors of Phytosciara and Bradysia are inferred to living plants habitat. Therefore, shifts in larval habitats from dead plant material to living plants may have occurred within the Sciaridae at least once. Based on the results, we discuss phylogenetic relationships within the family, and present an evolutionary scenario of development of larval habitats.     

Tsuga seed cones from the late Paleogene of southwestern China and their biogeographical and paleoenvironmental implications

Six Tsuga ovuliferous/seed cone impression fossils were discovered from the late Eocene (34.6 ± 0.8 Ma) Lawula Formation in Mangkang County, eastern Tibet and the early Oligocene (32 ± 1 Ma) lacustrine deposits in Lühe Basin, Nanhua County, Yunnan Province. These two fossil sites are both located in southwestern China, ∼800 km apart from each other. These fossils represent the oldest records of this genus in southwestern China, even earliest reliable macrofossil records of this genus in the world. These well-preserved seed cones provide sufficient materials for the establishment of Tsuga asiatica Wu et Zhou n. sp. to accommodate five specimens, leaving one to be assigned to T. cf. dumosa Eichler (cf. Wu et Zhou). Both qualitative and quantitative comparisons with other cone fossils and cones of all living species of the genus suggested that T. asiatica shares more similarities with one of the basal species of the genus T. heterophylla. The discovery of late Paleogene macrofossil records of Tsuga in southwestern China supports the previous hypothesis of the early disposal routes of this coniferous genus predicted by phylogenetic analysis. The elevation ranges and the climate requirements of living species that are closely related to our fossils suggest that the southeastern edge of the Qinghai-Tibetan Plateau should be much warmer, and wetter in late Paleogene than nowadays.     

Revisión sistemática de aceptación de la tecnología digital en personas mayores. Perspectiva de los modelos TAM

In this study, a systematic review of researches published about the determining factors in the intention of the use of devices and digital technology in older adults has been carried out. The technology acceptance model is the framework used. The main result of the analysis of 60 scientific articles published between 2010 and 2020 has been summarized after a bibliographic research and a subsequent selection process in the most important bibliographic databases: Web of Science (n = 45), Scopus (n = 66) and Google Scholar (n = 224). It has been proved the predominance of quantitative studies with a prevalence of study factors that associate the intention to use technology with: Facilitating conditions, experience, health, social support, emotions, perceived enjoyment, perceived satisfaction in using technology, anxiety, perceived risk, motivation, cost, subjective norm and social influence. The results of the systematic review allow us to conclude the urgent need to incorporate new research on the identification and influence of these factors that stand between older adults and technology, as possible barriers or facilitators of use.     

Alkaloids from the roots of Stemona javanica (Kunth) Engl. (Stemonaceae) and their anti-malarial,acetylcholinesterase inhibitory and cytotoxic activities

Two new protostemonine-type alkaloids, javastemonine A and B (3 and 4) have been isolated from the root extracts of Stemona javanica together with four known Stemona alkaloids, 13-demethoxy-11(S*),12(R*)-dihydroprotostemonine (1), isoprotostemonine (2), protostemonine and isomaistemonine. The structures and relative configurations of the new alkaloids were determined by spectroscopic analysis. The alkaloids 1 and 2 and protostemonine showed moderated antiplasmodial activities against the Plasmodium falciparum strains, TM4 (IC₅₀ values of 17.7 ± 3.7, 16.8 ± 5.4, 16.0 ± 4.2 μg/mL, respectively) and K1 (IC₅₀ values of 16.8 ± 3.1, 14.1 ± 3.7, 11.9 ± 3.3 μg/mL, respectively). These compounds showed no significant cytotoxicities against KB or Vero cells or acetylcholinesterase inhibitory activities.     

Synthesis,EPR, electrochemistry and EXAFS studies of ruthenium(III) complexes with a symmetrical tetradentate N2O2 Schiff base

A series of new hexa-coordinated ruthenium(III) complexes of the type [RuX(Nap-o-phd)(EPh₃)] (where, H₂-Nap-o-phd = N,N′-bis(2-hydroxy-1-naphthaldehyde) o-phenylene diamine; X = Cl or Br; E = P or As) have been prepared by reacting [RuX₃(EPh₃)₃] and [RuBr₃(PPh₃)₂(MeOH)] (where X = Cl or Br; E = P or As) with tetradentate Schiff base ligand (H₂-Nap-o-phd) in 1:1 molar ratio. The complexes have been characterized by elemental analyses, infra red, electronic, electron paramagnetic resonance spectroscopy and cyclic voltammetry. The coordination geometry and structure of the complexes have been investigated by extended X-ray absorption fine structure (EXAFS) spectroscopy and an octahedral structure has been proposed.     

Quaternary structure of KATP channel SUR2A nucleotide binding domains resolved by synchrotron radiation X-ray scattering

Heterodimeric nucleotide binding domains NBD1/NBD2 distinguish the ATP-binding cassette protein SUR2A, a recognized regulatory subunit of cardiac ATP-sensitive K⁺ (K_ATP) channels. The tandem function of these core domains ensures metabolism-dependent gating of the Kir6.2 channel pore, yet their structural arrangement has not been resolved. Here, purified monodisperse and interference-free recombinant particles were subjected to synchrotron radiation small-angle X-ray scattering (SAXS) in solution. Intensity function analysis of SAXS profiles resolved NBD1 and NBD2 as octamers. Implemented by ab initio simulated annealing, shape determination prioritized an oblong envelope wrapping NBD1 and NBD2 with respective dimensions of 168 × 80 × 37 Å³ and 175 × 81 × 37 Å³ based on symmetry constraints, validated by atomic force microscopy. Docking crystal structure homology models against SAXS data reconstructed the NBD ensemble surrounding an inner cleft suitable for Kir6.2 insertion. Human heart disease-associated mutations introduced in silico verified the criticality of the mapped protein–protein interface. The resolved quaternary structure delineates thereby a macromolecular arrangement of K_ATP channel SUR2A regulatory domains.