Lactose binding to human galectin-7 (p53-induced gene 1) induces long-range effects through the protein resulting in increased dimer stability and evidence for positive cooperativity.

16 pags, 11 figs, 3 tabs. -- Supplementary data for this article are available online at: http://glycob.oxfordjournals.org/lookup/suppl/doi:10.1093/glycob/cwt005/-/DC1The product of p53-induced gene 1 is a member of the galectin family, i.e., galectin-7 (Gal-7). To move beyond structural data by X-ray diffraction, we initiated the study of the lectin by nuclear magnetic resonance (NMR) and circular dichroism spectroscopies, and molecular dynamics (MD) simulations. In concert, our results indicate that lactose binding to human Gal-7 induces long-range effects (minor conformational shifts and changes in structural dynamics) throughout the protein that result in stabilization of the dimer state, with evidence for positive cooperativity. Monte Carlo fits of 15N-Gal-7 HSQC titrations with lactose using a two-site model yield K1 = 0.9 ± 0.6 × 103 M−1 and K2 = 3.4 ± 0.8 × 103 M−1. Ligand binding-induced stabilization of the Gal-7 dimer was supported by several lines of evidence: MD-based calculations of interaction energies between ligand-loaded and ligand-free states, gel filtration data and hetero-FRET spectroscopy that indicate a highly reduced tendency for dimer dissociation in the presence of lactose, CD-based thermal denaturation showing that the transition temperature of the lectin is significantly increased in the presence of lactose, and saturation transfer difference (STD) NMR using a molecular probe of the monomer state whose presence is diminished in the presence of lactose. MD simulations with the half-loaded ligand-bound state also provided insight into how allosteric signaling may occur. Overall, our results reveal long-range effects on Gal-7 structure and dynamics, which factor into entropic contributions to ligand binding and allow further comparisons with other members of the galectin family.This work was supported by a research grant from the National Institutes of Health (CA 096090 to K.H.M.), the RAS program “Molecular and Cellular Biology” RFBR grant (No. 12 04 31360 to E.E.), the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement no 260600 (“GlycoHIT”), grants CTQ2009-08536 and BFU2009-10052 and a FPI PhD fellowship to M.A.B. from the Spanish Ministry of Science and Innovation and the CIBER of Respiratory Diseases (CIBERES), an initiative from the Spanish Institute of Health Carlos III (ISCIII). E.E. was supported by a Travel Grant from the Minnesota Supercomputing Institute (University of Minnesota) during her stay in the research lab of Prof. K.H. Mayo. I.N. was supported in the Mayo lab by National Institutes of Health Hematology Training Grant (HL 07062

Cholera in Azov area

The purpose of research is analysis of clinical course and treatment results of patients with cholera in the Azov area. Materials and methods. During the period from 29.05.2011 to 19.08.2011 33 cases of cholera (32 adults and 1 child) and 25 vibrio carriers (22 adults and 3 children), which were caused by toxigenic strains of Vibrio cholera El Tor serogroup O1 Ogawa. Results. Likely factors of disease transmission in Mariupol are sea and river water, and the fish that were caught in the waters of the city. Typical and watery diarrhoea, vomiting, abdominal pain and lack of normal body temperature, dehydration syndrome, characterized clinical cholera for adults in most cases. The mean duration of diarrhoea was 6,6 days. At 46.9% observed atypical symptoms in 10 (31,3%) – abdominal pain (1 patient cramping in 7 cases, localized in the epigastria region, at 2-over stomach). In 5 patients (15,6%) had an increase in body temperature to 37,2–37,7 degrees Celsius. In 15 (46,9%) patients had severe nausea accompanied by vomiting. Easy for cholera was observed in 1 (3.1%), moderate – in 14 (43,8%), heavy – in 17 (53,1%) patients. Dehydration I level is set at 4 (12,5%), II – from 6 (18,7%), III – in 18 (56,3%), IV – 4 (12,5%) patients. Cholera outbreak was characterized by a predominance of severe disease and severe dehydration (III and IV), which was observed in 68.8% of patients. The decisive factor in the treatment of cholera patients was initiated in a timely manner rehydration therapy, in particular the introduction of the solution «Trisol». Against the background of rehydration therapy hyperkalaemia was observed in 9,4% of cases, vascular rehydration at 9,4%, the cell rehydration in 3,1% of patients. Fatal accidents cholera outbreaks have not been observed. Conclusion. Clinical diagnosis of cholera and the provision of medical care in the prehospital phase were poor, indicating the need for systematic conducting training seminars among experts of ambulance, as well as doctors of primary medical network. Cholera hospitals (or base hospitals) must be equipped with a mobile diagnostics (ultrasound, X-rays) and modern laboratory equipment. The required list of drug supply cholera hospitals should be expanded in view of possible complications and comorbidities

CHRONIC MYELOID LEUKEMIA: EPIDEMIOLOGY AND FIFTEENTH YEARS OF THERAPY IN THE NOVOSIBIRSK REGION

The incidence of chronic myeloid leukemia from 2004 to 2018 amounted to 0.63 per 100 000 people per year. The prevalence of chronic myeloid leukemia over the past 15 years has increased from 1.88 to 7.02 case per 100 000 people. We have analyzed the therapy outcomes of chronic myeloid leukemia of patients, received imatinib treatment for more than 12 month (165 patients). Complete hematologic response is attained in 88.5 % cases (146 patients), complete cytogenetic response (CCR) (ph+<0 %) – in 72.1 % cases (119 patients), major molecular response (MMR) – in 50.3 % cases (83 patients). The primary resistance to imatinib is observed in 21.8 % cases (36 patients), secondary – in 6.1 % cases (10 patients – 7 patients have lost the complete hematologic response and CCR and 3 patients have lost the CCR and MMR). The thirteen patients of those, who have primary or secondary resistance to imatinib have been treated with second-generation tyrosine kinase inhibitors (nilotinib, dasatinib) and in 93,3 % cases (14 patients) CCR have been obtained, 10 patients (66.7 %) have attained the CCR and 9 patients (60 %) have achieved MMR. Among all those patients treated with I and II generations tyrosine kinase inhibitors, we have carried out the overall survival (OS) analysis and obtained following results: the median of OS have not been achieved, 5-year OS rate is estimated as 90 %, 10-year OS rate – more than 77 %, calculated 15-year OS – more than 60 %

The genetic history of admixture across inner Eurasia

This is the author accepted manuscript. The final version is available from Nature Research via the DOI in this record.Data Availability. Genome-wide sequence data of two Botai individuals (BAM format) are available at the European Nucleotide Archive under the accession number PRJEB31152 (ERP113669). Eigenstrat format array genotype data of 763 present-day individuals and 1240K pulldown genotype data of two ancient Botai individuals are available at the Edmond data repository of the Max Planck Society (https://edmond.mpdl.mpg.de/imeji/collection/Aoh9c69DscnxSNjm?q=).The indigenous populations of inner Eurasia, a huge geographic region covering the central Eurasian steppe and the northern Eurasian taiga and tundra, harbor tremendous diversity in their genes, cultures and languages. In this study, we report novel genome-wide data for 763 individuals from Armenia, Georgia, Kazakhstan, Moldova, Mongolia, Russia, Tajikistan, Ukraine, and Uzbekistan. We furthermore report additional damage-reduced genome-wide data of two previously published individuals from the Eneolithic Botai culture in Kazakhstan (~5,400 BP). We find that present-day inner Eurasian populations are structured into three distinct admixture clines stretching between various western and eastern Eurasian ancestries, mirroring geography. The Botai and more recent ancient genomes from Siberia show a decrease in contribution from so-called “ancient North Eurasian” ancestry over time, detectable only in the northern-most “forest-tundra” cline. The intermediate “steppe-forest” cline descends from the Late Bronze Age steppe ancestries, while the “southern steppe” cline further to the South shows a strong West/South Asian influence. Ancient genomes suggest a northward spread of the southern steppe cline in Central Asia during the first millennium BC. Finally, the genetic structure of Caucasus populations highlights a role of the Caucasus Mountains as a barrier to gene flow and suggests a post-Neolithic gene flow into North Caucasus populations from the steppe.Max Planck SocietyEuropean Research Council (ERC)Russian Foundation for Basic Research (RFBR)Russian Scientific FundNational Science FoundationU.S. National Institutes of HealthAllen Discovery CenterUniversity of OstravaCzech Ministry of EducationXiamen UniversityFundamental Research Funds for the Central UniversitiesMES R

Холера в Приазовье

The purpose of research is analysis of clinical course and treatment results of patients with cholera in the Azov area. Materials and methods. During the period from 29.05.2011 to 19.08.2011 33 cases of cholera (32 adults and 1 child) and 25 vibrio carriers (22 adults and 3 children), which were caused by toxigenic strains of Vibrio cholera El Tor serogroup O1 Ogawa. Results. Likely factors of disease transmission in Mariupol are sea and river water, and the fish that were caught in the waters of the city. Typical and watery diarrhoea, vomiting, abdominal pain and lack of normal body temperature, dehydration syndrome, characterized clinical cholera for adults in most cases. The mean duration of diarrhoea was 6,6 days. At 46.9% observed atypical symptoms in 10 (31,3%) – abdominal pain (1 patient cramping in 7 cases, localized in the epigastria region, at 2-over stomach). In 5 patients (15,6%) had an increase in body temperature to 37,2–37,7 degrees Celsius. In 15 (46,9%) patients had severe nausea accompanied by vomiting. Easy for cholera was observed in 1 (3.1%), moderate – in 14 (43,8%), heavy – in 17 (53,1%) patients. Dehydration I level is set at 4 (12,5%), II – from 6 (18,7%), III – in 18 (56,3%), IV – 4 (12,5%) patients. Cholera outbreak was characterized by a predominance of severe disease and severe dehydration (III and IV), which was observed in 68.8% of patients. The decisive factor in the treatment of cholera patients was initiated in a timely manner rehydration therapy, in particular the introduction of the solution «Trisol». Against the background of rehydration therapy hyperkalaemia was observed in 9,4% of cases, vascular rehydration at 9,4%, the cell rehydration in 3,1% of patients. Fatal accidents cholera outbreaks have not been observed. Conclusion. Clinical diagnosis of cholera and the provision of medical care in the prehospital phase were poor, indicating the need for systematic conducting training seminars among experts of ambulance, as well as doctors of primary medical network. Cholera hospitals (or base hospitals) must be equipped with a mobile diagnostics (ultrasound, X-rays) and modern laboratory equipment. The required list of drug supply cholera hospitals should be expanded in view of possible complications and comorbidities.Цель исследования – анализ клинического течения и результатов лечения больных холерой в Приазовье. Материалы и методы. За период с 29.05.2011 г. по 19.08.2011 г. зарегистрировано 33 случая заболевания холерой (32 взрослых и 1 ребенок) и 25 вибрионосителей (22 взрослых и 3 детей), которые были вызваны токсигенными штаммами холерного вибриона Эль-Тор серогруппы O1 Огава. Результаты. Вероятными факторами передачи заболевания в г. Мариуполе являлись морская и речная вода, а также рыба, которая была выловлена в акватории города. Клиническое течение холеры у взрослых в большинстве случаев было типичным и характеризовалось водянистой диареей, рвотой, отсутствием болей в животе, нормальной температурой тела, синдромом обезвоживания. Средняя продолжительность диареи составила 6,6 суток. У 46,9% наблюдались нетипичные симптомы: у 10 (31,3%) – боль в животе (у 1 больного схваткообразная, в 7 случаях локализовалась в эпигастральной области, у 2 – по всему животу). У 5 пациентов (15,6%) отмечено повышение температуры тела до 37,2–37,7 °С. У 15 (46,9%) пациентов рвота сопровождалась выраженной тошнотой. Легкое течение холеры отмечено у 1 (3,1%), средней тяжести – у 14 (43,8%), тяжелое – у 17 (53,1%) больных. Дегидратация I степени установлена у 4 (12,5%), II – у 6 (18,7%), III – у 18 (56,3%), IV – у 4 (12,5%) пациентов. Вспышка холеры характеризовалась преобладанием тяжелых форм болезни и выраженной дегидратацией (III и IV степени), которая отмечена у 68,8% больных. Решающим фактором в терапии больных холерой была своевременно начатая регидратационная терапия, в частности, введение раствора «Трисоль». На фоне регидратационной терапии наблюдали гиперкалиемию в 9,4% случаев, сосудистую гипергидратацию у 9,4%, клеточную гипергидратацию у 3,1% больных. Летальные случаи на вспышке холеры не наблюдались. Заключение. Клиническая диагностика холеры и оказание лечебной помощи на догоспитальном этапе были неудовлетворительными, что свидетельствует о необходимости систематического проведения учебно-зачетных семинаров среди специалистов СМП, а также врачей первичной врачебной сети. Холерные госпитали (или базовые лечебные учреждения) должны быть оснащены передвижной диагностической (УЗИ, рентген) и современной лабораторной аппаратурой. Необходимый перечень лекарственного обеспечения холерных госпиталей следует расширить с учетом возможных осложнений и сопутствующих заболеваний. 

NMR Studies on Structure and Dynamics of the Monomeric Derivative of BS-RNase: New Insights for 3D Domain Swapping

Three-dimensional domain swapping is a common phenomenon in pancreatic-like ribonucleases. In the aggregated state, these proteins acquire new biological functions, including selective cytotoxicity against tumour cells. RNase A is able to dislocate both N- and C-termini, but usually this process requires denaturing conditions. In contrast, bovine seminal ribonuclease (BS-RNase), which is a homo-dimeric protein sharing 80% of sequence identity with RNase A, occurs natively as a mixture of swapped and unswapped isoforms. The presence of two disulfides bridging the subunits, indeed, ensures a dimeric structure also to the unswapped molecule. In vitro, the two BS-RNase isoforms interconvert under physiological conditions. Since the tendency to swap is often related to the instability of the monomeric proteins, in these paper we have analysed in detail the stability in solution of the monomeric derivative of BS-RNase (mBS) by a combination of NMR studies and Molecular Dynamics Simulations. The refinement of NMR structure and relaxation data indicate a close similarity with RNase A, without any evidence of aggregation or partial opening. The high compactness of mBS structure is confirmed also by H/D exchange, urea denaturation, and TEMPOL mapping of the protein surface. The present extensive structural and dynamic investigation of (monomeric) mBS did not show any experimental evidence that could explain the known differences in swapping between BS-RNase and RNase A. Hence, we conclude that the swapping in BS-RNase must be influenced by the distinct features of the dimers, suggesting a prominent role for the interchain disulfide bridges

Structural Biology by NMR: Structure, Dynamics, and Interactions

The function of bio-macromolecules is determined by both their 3D structure and conformational dynamics. These molecules are inherently flexible systems displaying a broad range of dynamics on time-scales from picoseconds to seconds. Nuclear Magnetic Resonance (NMR) spectroscopy has emerged as the method of choice for studying both protein structure and dynamics in solution. Typically, NMR experiments are sensitive both to structural features and to dynamics, and hence the measured data contain information on both. Despite major progress in both experimental approaches and computational methods, obtaining a consistent view of structure and dynamics from experimental NMR data remains a challenge. Molecular dynamics simulations have emerged as an indispensable tool in the analysis of NMR data

13C multiplet nuclear magnetic resonance relaxation-derived ring puckering and backbone dynamics in proline-containing glycine-based peptides.

13CH2-multiplet nuclear magnetic resonance relaxation studies on proline (P)-containing glycine (G)-based peptides, GP, PG, GPG, PGG, and GPGG, provided numerous dipolar auto- and cross-correlation times for various motional model analyses of backbone and proline-ring bond rotations. Molecular dynamics simulations and bond rotation energy profiles were calculated to assess which motions could contribute most to observed relaxation phenomena. Results indicate that proline restricts backbone psi 1, psi 2, and phi 2 motions by 50% relative to those found for a polyglycine control peptide. psi 1 rotations are more restricted in the trans-proline isomer state than in the cis form. A two-state jump model best approximates proline ring puckering which in water could occur either by the C gamma endo-exo or by the C2 interconversion mechanism. The temperature dependence (5 degrees to 75 degrees C) of C beta, and C gamma, and C delta angular changes is rather flat, suggesting a near zero enthalpic contribution to the ring puckering process. In lower dielectric solvents, dimethylsulfoxide and methanol, which may mimic the hydrophobic environment within a protein, the endo-exo mechanism is preferred