Hybrid inorganic-organic capsules for efficient intracellular delivery of novel siRNAs against influenza A (H1N1) virus infection

This work was supported by ARUK project grant 21210 ‘Sustained and Controllable Local Delivery of Anti-inflammatory Therapeutics with Nanoengineered Microcapsules’. The work was also supported in part by Russian Foundation of Basic Research grants No. 16-33-50153 mol_nr, No. 16-33-00966 mol_a, Russian Science Foundation grant No. 15-15-00170 and Russian Governmental Program ‘‘Nauka’’, No. 1.1658.2016, 4002

Диагностика и лечение рака ротоглотки в реальной клинической практике в Республике Башкортостан: анализ за 2020 год

Introduction. Th e growing incidence of oropharyngeal cancer is driven by an increase in frequency of HPV-associated oropharyngeal cancer. Th e morbidity pattern of oropharyngeal cancer is area-specific.Aim. To analyze the oropharyngeal cancer diagnosis and treatment in the Republic of Bashkortostan for 2020.Materials and methods. Th e authors carried out a 2020 retrospective analysis of the diagnosis and treatment results of patients with oropharyngeal cancer. 79 patients were identified with this diagnosis. 84.8% (67/79) among them were males and 15.2% females (12/79). Th e mean age of the patients was 59.1 years. Th e site of primary tumor was on the oropharynx lateral wall in 37.8% cases (30/79), in the tongue root area — 24.1% (19/79), in the tonsils area — 17.7% (14/79), on the soft palate — 16.5% (13/79), on the oropharynx posterior wall — 3.8% (3/79).Results. Examination of tumor morphological types revealed squamous cell carcinoma (SCC) with various degrees of differentiation in 92.4% cases (73/79), adenocarcinoma of minor salivary gland — in 6.3% (5/79) and sarcoma in 1.2% (1/79). 57.5% of 73 patients with SCC (42/73) underwent protein (p16) immunohistochemistry, while 42.5% of the patients (31/73) did not. According to a surrogate marker for HPV, the following results were obtained for 42 patients: p16-positive in 23.8% cases (10/42), p16-negative in 76.2% (32/42). Stage distribution according to TNM-7: stage I — 11.4% (9/79), stage II — 17.7% (14/79), stage III — 36.7% (29/79), stage IV — 46.8% (37/79). Stage distribution according to TNM-8 (patients who underwent p16 immunohistochemistry): stage I — 11.9% (5/42), stage II — 23.8% (10/42), stage III — 19% (8/42), stage IV — 45.2% (19/42). In 2020, 72% of patients (57/79) received definitive treatment, 10.1% (8/79) — palliative care, 15.2% (12/79) — supportive care, and 2.5% (2/79) refused medical treatment.Discussion. Th e various types of radiation therapy were used as the main defi nitive treatment for patients with oropharyngeal cancer in 69.2% cases (45/65). Only 18.5% of patients (12/65) underwent surgery, 58.3% of which (7/12) received post-surgery radiation therapy.Conclusion. 57.5% of patients (42/73) were detected with HPV status, 23.8% (10/42) revealed surrogate markers for HPV association. 69.2% of patients (45/65) received radiation therapy as the definitive treatment. 18.5% of patients (12/65) underwent surgery, 58.3% of which (7/12) received postsurgery radiation therapy.Введение. Современная тенденция роста заболеваемости раком ротоглотки обусловлена ростом ВПЧ-ассоциированной формы рака ротоглотки. Структура данной заболеваемости имеет территориальные особенности.Цель исследования. Провести анализ диагностики и лечения рака ротоглотки в Республике Башкортостан за 2020 год.Материалы и методы. Выполнен ретроспективный анализ результатов диагностики и лечения пациентов с диагнозом «рак ротоглотки» за 2020 год. Данный диагноз был установлен 79 пациентам. Доля пациентов мужского пола составила 84,8 % (67/79), женского пола — 15,2 % (12/79). Средний возраст пациентов — 59,1 года. Первичная опухоль локализовалась на боковой стенке ротоглотки в 37,8 % (30/79), в области корня языка — 24,1 % (19/79), в области миндалин — 17,7 % (14/79), мягкого неба — 16,5 % (13/79), на задней стенке ротоглотки — 3,8 % (3/79) случаев. Результаты. При анализе морфологических форм опухоли в 92,4 % (73/79) случаев это плоскоклеточный рак (ПКР) различной степени дифференцировки, в 6,3 % (5/79) — аденокарциномы из малых слюнных желез, в 1,2 % (1/79) — саркома. Из 73 пациентов с ПКР ротоглотки иммуногистохимическое исследование на белок р16 было выполнено 57,5 % (42/73) пациентов, 42,5 % (31/73) пациентов не проведено. У 42 пациентов по данным суррогатного маркера ВПЧ-ассоциации получены следующие результаты: р16-положительный результат у 23,8 % (10/42), р16-отрицательный результат у 76,2 % (32/42). Распределение по стадиям в соответствии TNM- 7: 1-я стадия — 11,4 % (9/79), 2-я стадия — 17,7 % (14/79), 3-я стадия — 36,7 % (29/79), 4-я стадия — 46,8 % (37/79). Распределение по стадиям в соответствии TNM-8 (пациентам, которым выполнено исследование ИГХ на р16): 1-я стадия — 11,9 % (5/42), 2-я стадия — 23,8 % (10/42), 3-я стадия — 19 % (8/42), 4-я стадия — 45,2 % (19/42). За 2020 год радикальное лечение проведено 72 % (57/79), паллиативное лечение — 10,1 % (8/79), симптоматическая терапия — 15,2 % (12/79), от лечения отказались 2,5 % (2/79) пациентов.Обсуждение. Основным радикальным методом лечения пациентов с раком ротоглотки была лучевая терапия в различных вариантах — 69,2 % (45/65) случаев. Хирургическое лечение проведено только 18,5 % (12/65) пациентов, из них 58,3 % (7/12) проведена послеоперационная лучевая терапия.Заключение. Определение ВПЧ-статуса было выполнено у 57,5 % (42/73), и у 23,8 % (10/42) пациентов выявлены суррогатные маркеры ВПЧ-ассоциации. 69,2 % (45/65) пациентов в качестве основного радикального лечения была проведена лучевая терапия в различных вариантах. 18,5 % (12/65) пациентов было проведено хирургическое лечение, из них 58,3 % (7/12) проведена послеоперационная лучевая терапия

A method for estimation of drug affinity constants to the open conformational state of calcium channels.

The affinity of D600 to calcium channels in the open state has been examined in isolated smooth muscle cells of the rabbit ear artery. Calcium channel currents were measured in high external barium solution by means of the patch-clamp technique. The current inhibition in various D600 concentrations (3-100 microM) on application of trains of short test pulses (20-80 ms) has been studied in nonmodified calcium channels and in cells where the calcium channels were modified by the agonist dihydropyridine (+) 202,791 (100 nM). The kinetics of the peak current decay has been analyzed with a mathematical model which is based on the experimental finding that D600 interacts primarily with calcium channels in the open conformational state. The model approach allows the estimation of drug affinity constants of D600 to the calcium channel in the open conformation. An association rate constant to the open conformational state of D600 of 6.16 x 10(4) M-1 s-1 was estimated. The association rate of the drug was not significantly changed after the calcium channels have been modified with 100 nM (+) 202,791. A method for correction of rate constants for possible drug trapping is discussed

Different Inward and Outward Conduction Mechanisms in Na_VMs Suggested by Molecular Dynamics Simulations

<div><p>Rapid and selective ion transport is essential for the generation and regulation of electrical signaling pathways in living organisms. Here, we use molecular dynamics (MD) simulations with an applied membrane potential to investigate the ion flux of bacterial sodium channel Na_VMs. 5.9 µs simulations with 500 mM NaCl suggest different mechanisms for inward and outward flux. The predicted inward conductance rate of ∼27±3 pS, agrees with experiment. The estimated outward conductance rate is 15±3 pS, which is considerably lower. Comparing inward and outward flux, the mean ion dwell time in the selectivity filter (SF) is prolonged from 13.5±0.6 ns to 20.1±1.1 ns. Analysis of the Na⁺ distribution revealed distinct patterns for influx and efflux events. In 32.0±5.9% of the simulation time, the E53 side chains adopted a flipped conformation during outward conduction, whereas this conformational change was rarely observed (2.7±0.5%) during influx. Further, simulations with dihedral restraints revealed that influx is less affected by the E53 conformational flexibility. In contrast, during outward conduction, our simulations indicate that the flipped E53 conformation provides direct coordination for Na⁺. The free energy profile (potential of mean force calculations) indicates that this conformational change lowers the putative barriers between sites S_CEN and S_HFS during outward conduction. We hypothesize that during an action potential, the increased Na⁺ outward transition propensities at depolarizing potentials might increase the probability of E53 conformational changes in the SF. Subsequently, this might be a first step towards initiating slow inactivation.</p></div

Annotations of the SF ion interacting site.

<p>SF backbone atoms are shown as blue sticks, E53 and S54 side chain atoms are also shown to determine the site S_HFS (only two opposite subunits are shown for clarity). In addition, sodium ions are depicted with yellow spheres. Sites S_EX (−5.00≤Z<0.00 Å), S_HFS (0.00≤Z<2.75 Å), S_BAR (2.75≤Z<4.75 Å), site S_CEN (4.75≤Z<7.75 Å) and site S_IN (7.75≤Z<10.25 Å) and the length of each site were annotated at the lateral sides of the figure.</p

Outward ionic binding modes and PMF.

<p>A) Overlay of the conformational space of the probe ions (yellow), coupling sodium ions (blue) and E53 carboxyl oxygen distributions (transparent spheres with different intensity) for different ionic binding modes at different interaction sites. A snapshot of a typical E53 sidechain conformation is shown as stick representation (red, carboxyl oxygens; yellow, sidechain carbons), I–V are non-flip binding modes and II′–IV′ are flipping binding modes for sites S_HFS, S_BAR and S_CEN. Arrows indicate the direction of ion conduction; B) 1-D PMF of outward conduction in SF region, the largest free energy barriers are labeled by terminal peaks and wells, corresponding positions of typical binding modes on the energy profile are labeled. Error estimations shown in the figure are S.E.M.</p

Influences of different flipping states at Δq = 4<i>e</i> (n = 4).

<p>A) Average ion flux counts through the SF over time of inward conduction (color: blue). The simulations without restraints of E53 are depicted as solid line, the ones with the “one-flip” restraints are shown as dotted line and the ones with the “non-flip” restraints are shown as dashed line. B) Average ion flux count through the SF over time of outward conduction (color: red). Error estimations shown in the figure are S.E.M.</p

Transmembrane voltages comparison.

<p>A) Electrostatic potential across the simulation box calculated from the simulations without dihedral restraints on E53. B) Electrostatic potential across the simulation box calculated from the snapshots (t>5 ns) extracted from the simulation in <a href="http://www.ploscompbiol.org/article/info:doi/10.1371/journal.pcbi.1003746#pcbi-1003746-g002" target="_blank">Figure 2</a>A without ion conduction events in neither inward nor outward directions. C) Electrostatic potential across the simulation box calculated from the “no salt” simulations. (Error estimations shown in the figure are S.D.).</p

Ion flux in double bilayer simulations without dihedral restraints on E53 (n = 4).

<p>A) Average ion flux count of inward conduction through the SF over time (color: blue). B) Average ion flux count of outward conduction through the SF over time (color: red). Error estimations shown in the figure are S.E.M.</p

PMF comparison for outward conduction.

<p>A) PMF of the simulations without restraints of E53 is depicted as solid line. (B) PMF of the simulations with “one-flip” restraints. C) PMF of the simulations with “non-flip” restraints. The largest free energy barriers in each figure are labeled by terminal peaks and wells. Error estimations shown in the figure are S.E.M.</p