The Origin of Minus-end Directionality and Mechanochemistry of Ncd Motors

Adaptation of molecular structure to the ligand chemistry and interaction with the cytoskeletal filament are key to understanding the mechanochemistry of molecular motors. Despite the striking structural similarity with kinesin-1, which moves towards plus-end, Ncd motors exhibit minus-end directionality on microtubules (MTs). Here, by employing a structure-based model of protein folding, we show that a simple repositioning of the neck-helix makes the dynamics of Ncd non-processive and minus-end directed as opposed to kinesin-1. Our computational model shows that Ncd in solution can have both symmetric and asymmetric conformations with disparate ADP binding affinity, also revealing that there is a strong correlation between distortion of motor head and decrease in ADP binding affinity in the asymmetric state. The nucleotide (NT) free-ADP (?-ADP) state bound to MTs favors the symmetric conformation whose coiled-coil stalk points to the plus-end. Upon ATP binding, an enhanced flexibility near the head-neck junction region, which we have identified as the important structural element for directional motility, leads to reorienting the coiled-coil stalk towards the minus-end by stabilizing the asymmetric conformation. The minus-end directionality of the Ncd motor is a remarkable example that demonstrates how motor proteins in the kinesin superfamily diversify their functions by simply rearranging the structural elements peripheral to the catalytic motor head domain

Diagnostics of testicular torsion using the TWIST scale (Testicular Workup for Ischemia and Suspected Torsion)

Introduction. Testicular torsion (TT) is the most common pediatric emergency urological pathology. To reduce the duration of the diagnostic stage, systems for assessing the risks of testicular torsion based on anamnesis and clinical symptoms were proposed. In 2013, Barbosa et al. proposed the TWIST system (testicular examination for ischemia and suspected torsion), which became the most well-known and widespread. This system makes it possible to identify groups of patients who do not require scrotal ultrasound, which reduces the number of stages in the diagnosis of TT.Objective. To evaluate the experience of using and diagnostic significance of the TWIST scale based on available data in scientific publications.Materials & methods. Review and analysis of literature data on the use of the TWIST scale.Results. We conducted an analysis of 13 publications, in which the results of using TWIST with statistical analysis were published. In all articles, the final diagnosis was established according to Doppler scrotal ultrasound or intraoperatively. Analysis of publications shows that even in large foreign medical centers there is a problem of emergency scrotal ultrasound, which increases the time of testicular ischemia with ТТ. To use the TWIST scale, only history and physical examination data are needed. Any specialist can use the scale in his practise. The low probability of TT in the low-risk group makes it possible not to perform routine scrotal revision, and, consequently, material and human resources are saved.Conclusion. Literature analysis has shown that the use of the original TWIST scale proposed by J.A. Barbosa, in case of suspected testicular torsion, has sufficient diagnostic accuracy, high sensitivity and specificity of TT detection, which significantly reduces the need for ultrasound, reduces the diagnostic time before surgery, that increases testicular survival

Minimally invasive ureteral reimplantation: trends in regions of the Russian Federation and The Republic of Belarus. Multi-center study

Introduction. Until recently, open ureteral reimplantation was considered the gold standard for the surgical treatment of ureterovesical junction (UVJ) pathologies. The introduction of minimally invasive ureteral reimplantation (MIUR) for the treatment of vesicoureteral reflux (VUR) and primary obstructive megaureter (POM) in children started in the 1990s. However, studies describing national trends in the use of minimally invasive and open approaches in the surgical treatment of UVJ pathology in children are limited.Objective. To describe changes in the use of MIUR and open ureteral reimplantation (OUR) between 2007 and 2022 in some regions of the Russian Federation and the Republic of Belarus and compare the results and complication rates of the two surgical approaches. Materials & methods. The study includes 1273 patients (1793 ureters), operated on for UVJ pathology in the period from 2007 to 2022. MIR was performed in 1356 (75.6%) ureters (913 for VUR and 443 for POM). The studied parameters included an annual amount of reimplantation, the age of patients, the frequency of intra- and postoperative complications, as well as the medium-term results of operations.Results. The use of MIUR techniques has increased significantly over time, and in 2022, seventy-five percent of surgeries were performed using MIUR. Analysis of the rate of adoption of MIUR by clinics showed that those that began using it first experienced a significant increase in frequency of use after 4 to 6 years, while those that started later took 2 to 3 years to achieve a significant positive trend. There were 5 (0.4%) intraoperative complications in the MIUR group. All these complications were classified as grade I according to the Satava grading system. There was no significant difference between MIUR and OUR in terms of postoperative complication rates (6.6% vs. 7.6%, p = 0.8). The efficiency of reimplantation was 96.6% in the MIUR group compared to 95.9% in the OUR group for POM and 96.2% in the MIR group compared to 94.6% in the OUR for VUR.Conclusion. In the regions of Russia involved in the study, there has been a trend towards completely replacing open surgery with minimally invasive techniques. Regarding complication rates and efficacy, MIUR is not significantly different from OUR

Origins and genetic legacy of prehistoric dogs

Dogs were the first domestic animal, but little is known about their population history and to what extent it was linked to humans. We sequenced 27 ancient dog genomes and found that all dogs share a common ancestry distinct from present-day wolves, with limited gene flow from wolves since domestication but substantial dog-to-wolf gene flow. By 11,000 years ago, at least five major ancestry lineages had diversified, demonstrating a deep genetic history of dogs during the Paleolithic. Coanalysis with human genomes reveals aspects of dog population history that mirror humans, including Levant-related ancestry in Africa and early agricultural Europe. Other aspects differ, including the impacts of steppe pastoralist expansions in West and East Eurasia and a near-complete turnover of Neolithic European dog ancestry

European registry on the management of helicobacter pylori infection (HP-EUREG protocol): The first results of Russian centers

Aim: To assess the clinical practice of diagnosis and treatment in patients with Helicobacter pylori infection and to compare this practice with the international guidelines in the European Registry on the management of Helicobacter pylori infection, Hp-EuReg protocol), a multicenter prospective observational study initiated by the European Helicobacter and Microbiota Study Group. Materials and methods: The data of 813 patients infected with H. pylori and entered in the Hp-EuReg register by the Russian centers in 2013-2015 were analyzed. Results: The most common methods for the primary diagnosis of H. pylori infection are histology (40.3%), rapid urease test (35.7%), and serology (17.2%). The duration of H. pylori eradication therapy was 7, 10, and 14 days in 18.0, 49.3, and 25.1%, respectively. To monitor the effectiveness of treatment, the investigators used a histological examination (34%), a urea breath test (27.3%), H. pylori stool antigen (22.8%), and a rapid urease test (16.3%). A serological test was carried out in 2.5% of the cases. No monitoring was done in 13.5% of the patients. The average eradication efficiency was 82.6%. If the therapy was ineffective, 80% of physicians did not intend to prescribe a new cycle of treatment. Conclusion: Significant differences were found between clinical practice and the current guidelines

Resistance of Renal Cell Carcinoma to Sorafenib Is Mediated by Potentially Reversible Gene Expression

Purpose: Resistance to antiangiogenic therapy is an important clinical problem. We examined whether resistance occurs at least in part via reversible, physiologic changes in the tumor, or results solely from stable genetic changes in resistant tumor cells. Experimental Design: Mice bearing two human RCC xenografts were treated with sorafenib until they acquired resistance. Resistant 786-O cells were harvested and reimplanted into naïve mice. Mice bearing resistant A498 cells were subjected to a 1 week treatment break. Sorafenib was then again administered to both sets of mice. Tumor growth patterns, gene expression, viability, blood vessel density, and perfusion were serially assessed in treated vs control mice. Results: Despite prior resistance, reimplanted 786-O tumors maintained their ability to stabilize on sorafenib in sequential reimplantation steps. A transcriptome profile of the tumors revealed that the gene expression profile of tumors upon reimplantation reapproximated that of the untreated tumors and was distinct from tumors exhibiting resistance to sorafenib. In A498 tumors, revascularization was noted with resistance and cessation of sorafenib therapy and tumor perfusion was reduced and tumor cell necrosis enhanced with re-exposure to sorafenib. Conclusions: In two RCC cell lines, resistance to sorafenib appears to be reversible. These results support the hypothesis that resistance to VEGF pathway therapy is not solely the result of a permanent genetic change in the tumor or selection of resistant clones, but rather is due to a great extent to reversible changes that likely occur in the tumor and/or its microenvironment

Processive Movement by a Kinesin Heterodimer with an Inactivating Mutation in One Head†

ABSTRACT: A single molecule of the motor enzyme kinesin-1 keeps a tight grip on its microtubule track, making tens or hundreds of discrete, unidirectional 8 nm steps before dissociating. This high duty ratio processive movement is thought to require a mechanism in which alternating stepping of the two head domains of the kinesin dimer is driven by alternating, overlapped cycles of ATP hydrolysis by the two heads. The R210K point mutation in Drosophila kinesin heavy chain was reported to disrupt the ability of the enzyme active site to catalyze ATP P-O bond cleavage. We expressed R210K homodimers as well as isolated R210K heads and confirmed that both are essentially inactive. We then coexpressed tagged R210K subunits with untagged wild-type subunits and affinity purified R210K/wild-type heterodimers together with the inactive R210K homodimers. In contrast to the R210K head or homodimer, the heterodimer was a highly active (>50 % of wild-type) microtubule-stimulated ATPase, and the heterodimer displayed high duty ratio processive movement in single-molecule motility experiments. Thus, dimerization of a subunit containing the inactivating mutation with a functional subunit can complement the mutation; this must occur either by lowering or by bypassing kinetic barriers in the ATPase or mechanical cycles of the mutant head. The observations provide support for kinesin-1 gating mechanisms in which one head stimulates the rate of essential processes in the other

Population genomics of post-glacial western Eurasia.

Western Eurasia witnessed several large-scale human migrations during the Holocene <sup>1-5</sup> . Here, to investigate the cross-continental effects of these migrations, we shotgun-sequenced 317 genomes-mainly from the Mesolithic and Neolithic periods-from across northern and western Eurasia. These were imputed alongside published data to obtain diploid genotypes from more than 1,600 ancient humans. Our analyses revealed a 'great divide' genomic boundary extending from the Black Sea to the Baltic. Mesolithic hunter-gatherers were highly genetically differentiated east and west of this zone, and the effect of the neolithization was equally disparate. Large-scale ancestry shifts occurred in the west as farming was introduced, including near-total replacement of hunter-gatherers in many areas, whereas no substantial ancestry shifts happened east of the zone during the same period. Similarly, relatedness decreased in the west from the Neolithic transition onwards, whereas, east of the Urals, relatedness remained high until around 4,000 BP, consistent with the persistence of localized groups of hunter-gatherers. The boundary dissolved when Yamnaya-related ancestry spread across western Eurasia around 5,000 BP, resulting in a second major turnover that reached most parts of Europe within a 1,000-year span. The genetic origin and fate of the Yamnaya have remained elusive, but we show that hunter-gatherers from the Middle Don region contributed ancestry to them. Yamnaya groups later admixed with individuals associated with the Globular Amphora culture before expanding into Europe. Similar turnovers occurred in western Siberia, where we report new genomic data from a 'Neolithic steppe' cline spanning the Siberian forest steppe to Lake Baikal. These prehistoric migrations had profound and lasting effects on the genetic diversity of Eurasian populations

Analysis of large versus small dogs reveals three genes on the canine X chromosome associated with body weight, muscling and back fat thickness

International audienceDomestic dog breeds display significant diversity in both body mass and skeletal size, resulting from intensive selective pressure during the formation and maintenance of modern breeds. While previous studies focused on the identification of alleles that contribute to small skeletal size, little is known about the underlying genetics controlling large size. We first performed a genome-wide association study (GWAS) using the Illumina Canine HD 170,000 single nucleotide polymorphism (SNP) array which compared 165 large-breed dogs from 19 breeds (defined as having a Standard Breed Weight (SBW) >41 kg [90 lb]) to 690 dogs from 69 small breeds (SBW ≤41 kg). We identified two loci on the canine X chromosome that were strongly associated with large body size at 82–84 megabases (Mb) and 101–104 Mb. Analyses of whole genome sequencing (WGS) data from 163 dogs revealed two indels in the Insulin Receptor Substrate 4 (IRS4) gene at 82.2 Mb and two additional mutations, one SNP and one deletion of a single codon, in Immunoglobulin Superfamily member 1 gene (IGSF1) at 102.3 Mb. IRS4 and IGSF1 are members of the GH/IGF1 and thyroid pathways whose roles include determination of body size. We also found one highly associated SNP in the 5’UTR of Acyl-CoA Synthetase Long-chain family member 4 (ACSL4) at 82.9 Mb, a gene which controls the traits of muscling and back fat thickness. We show by analysis of sequencing data from 26 wolves and 959 dogs representing 102 domestic dog breeds that skeletal size and body mass in large dog breeds are strongly associated with variants within IRS4, ACSL4 and IGSF1