Smallest near-infrared fluorescent protein evolved from cyanobacteriochrome as versatile tag for spectral multiplexing

From a single domain of cyanobacteriochrome (CBCR) we developed a near-infrared (NIR) fluorescent protein (FP), termed miRFP670nano, with excitation at 645 nm and emission at 670 nm. This is the first CBCR-derived NIR FP evolved to efficiently bind endogenous biliverdin chromophore and brightly fluoresce in mammalian cells. miRFP670nano is a monomer with molecular weight of 17 kDa that is 2-fold smaller than bacterial phytochrome (BphP)-based NIR FPs and 1.6-fold smaller than GFP-like FPs. Crystal structure of the CBCR-based NIR FP with biliverdin reveals a molecular basis of its spectral and biochemical properties. Unlike BphP-derived NIR FPs, miRFP670nano is highly stable to denaturation and degradation and can be used as an internal protein tag. miRFP670nano is an effective FRET donor for red-shifted NIR FPs, enabling engineering NIR FRET biosensors spectrally compatible with GFP-like FPs and blue-green optogenetic tools. miRFP670nano unlocks a new source of diverse CBCR templates for NIR FPs.Peer reviewe

Figure of Borromean knot in Lacanian structural psychoanalysis

The article deals the interpretations of some Lacanian terms and the principles of structural psychoanalysis.В статье рассмотрены интерпретации некоторых лакановских терминов и принципы структурного психоанализа

Intuition in the medical profession

Each person understands the meaning of the term intuition in their own way. For some, intuition is something elusive, intangible, indefinable, but they answer without a doubt that they would like to develop it in themselves. For others, it is an inner voice. Many people define intuition as something supernatural, mystical, descended from above, from somewhere outside, as a kind of “sixth sense”. We can talk about intuition as an instant and immediate insight based on the already existing background knowledge. Among the various versions of the meaning of the word “intuition”, we can distinguish the reference one, given in the “Explanatory Dictionary of the Russian Language” by S. I. Ozhegov: INTUITION, 1. Intuition, subtle understanding, insight into the very essence of something. 2. Direct comprehension of the truth without justification by evidence.Каждый человек по-своему понимает значение термина интуиция. Для одних интуиция – это что-то неуловимое, неосязаемое, неопределяемое, но они без сомнений отвечают, что хотели бы в себе её развить. Для других — это внутренний голос. Многие определяет интуицию как нечто сверхъестественное, мистическое, снизошедшее сверху, откуда-то извне, как своеобразное “шестое чувство”. Можно говорить об интуиции, имея в виду инсайт, мгновенное и непосредственное озарение на основе уже имеющихся фоновых знаний. Среди разнообразных версий значения слова “интуиция” можно выделить эталонное, приведенное в “Толковом словаре русского языка” С.И. Ожегова: ИНТУИЦИЯ, 1. Чутьё, тонкое понимание, проникновение в самую суть чего-нибудь. 2. Непосредственное, без обоснования доказательствами постижение истины

Application of isothermal titration calorimetry in evaluation of protein–nanoparticle interactions

Nanoparticles (NPs) offer a number of advantages over small organic molecules for controlling protein behaviour inside the cell. Protein binding to the surface of NPs depends on their surface characteristics, composition and method of preparation (Mandal et al. in J Hazard Mater 248–249:238–245, 2013). It is important to understand the binding affinities, stoichiometries and thermodynamical parameters of NP–protein interactions in order to see which interaction will have toxic and hazardous consequences and thus to prevent it. On the other side, because proteins are on the brink of stability, they may experience interactions with some types of NPs that are strong enough to cause denaturation or significantly change their conformations with concomitant loss of their biological function. Structural changes in the protein may cause exposure of new antigenic sites, “cryptic” peptide epitopes, potentially triggering an immune response which can promote autoimmune disease (Treuel et al. in ACS Nano 8(1):503–513, 2014). Mechanistic details of protein structural changes at NP surface have still remained elusive. Understanding the formation and persistence of the protein corona is critical issue; however, there are no many analytical methods which could provide detailed information about the NP–protein interaction characteristics and about protein structural changes caused by interactions with nanoparticles. The article reviews recent studies in NP–protein interactions research and application of isothermal titration calorimetry (ITC) in this research. The study of protein structural changes upon adsorption on nanoparticle surface and application of ITC in these studies is emphasized. The data illustrate that ITC is a versatile tool for evaluation of interactions between NPs and proteins. When coupled with other analytical methods, it is important analytical tool for monitoring conformational changes in proteins

Modular assembly of proteins on nanoparticles

Generally, the high diversity of protein properties necessitates the development of unique nanoparticle bio-conjugation methods, optimized for each different protein. Here we describe a universal bio-conjugation approach which makes use of a new recombinant fusion protein combining two distinct domains. The N-terminal part is Glutathione S-Transferase (GST) from Schistosoma japonicum, for which we identify and characterize the remarkable ability to bind gold nanoparticles (GNPs) by forming gold–sulfur bonds (Au–S). The C-terminal part of this multi-domain construct is the SpyCatcher from Streptococcus pyogenes, which provides the ability to capture recombinant proteins encoding a SpyTag. Here we show that SpyCatcher can be immobilized covalently on GNPs through GST without the loss of its full functionality. We then show that GST-SpyCatcher activated particles are able to covalently bind a SpyTag modified protein by simple mixing, through the spontaneous formation of an unusual isopeptide bond

Psychosomatic correlation with hypertension complicated by acute violation of cerebral circulation

The objective was to study the gender Impact of emotional factors on the performance of total metabolism, cardiovascular remodeling in patients with arterial hypertension complicated with ischemic stroke. The study included 64 patient hospitalized in neurological department with arterial hypertension complicated by acute violation of cerebral blood flow by ischemic type I and II degrees. 32 of them (mean aged 48.3 ± 10.4 years) and 32 women (mean aged 51.3 ± 14.0 years). All patient study of lipid fasting venous blood indicators, ultrasound of the heart and blood vessels, blood pressure measurement according to standard procedure, as well as questioning by using psychological questionnaires. Number and nature of the identified associations between total metabolism, cardiovascular remodeling and psychological factors depend on gender. Identified correlations can be useful in the planning of programmes of rehabilitation and secondary prevention of stroke.В исследование включено 64 пациента, госпитализированных в неврологическое отделение для больных ОНМК МБУЗ ГКБ № 8 г. Челябинска с артериальной гипертензией, осложненной острым наррением мозгового кровообращения по ишемическому типу I и II степени тяжести. Из них - 32 мужчины (средний возраст 48,3±10,4 лет) и 32 женщины (средний возраст 51,3±14,0 лет). Всем обследуемым проводилось исследование липидных показателей венозной крови натощак, ультразвуковое исследование сердца и сосудов, измерение артериального давления по стандартной процедуре, а также анкетирование с помощью психологических опросников. Количество и характер выявленных ассоциаций между показателями липопротеинового обмена, кардиоваскулярного ремоделирования и психологическими факторами зависел от пола. Выявленные корреляции могут оказаться полезными при планировании программ вторичной профилактики и реабилитации инсульта

Эффективность и безопасность кабозантиниба у пациентов с распространенным почечно-клеточным раком: российское многоцентровое наблюдательное исследование

Purpose: an assessment of efficacy and safety of cabozantinib in unselected patients with metastatic renal cell carcinoma in the first and subsequent lines of therapy.Materials and methods. Russian multicenter observational study included 92 consecutive patients with morphologically verified metastatic renal cell carcinoma treated with cabozantinib (60 mg/d) in 16 Russian centers. Median age of the patients was 56 (19-79) years, a male-to-female ratio - 3:1. At the start of cabozantinib therapy 27.2 % of patients had ECOG PS 2. Most common histological type of kidney cancer was clear-cell RCC (90.2 %). Most patients were diagnosed with synchronous (71.7 %) multiple metastases (60.9 %). Previous nephrectomy was performed in 87.0 % of cases. Prognosis according to International Metastatic Renal Cancer Database Consortium (IMDC) score was assessed as favorable in 5.4 %, intermediate - in 58.7 % and poor - in 35.9 % patients. Cabozantinib as the first-line therapy was administered in 9 (9.8 %), following 1-5 lines of systemic treatment - in 83 (90.2 %) cases. Median follow-up was 11 (2.3-44.5) months.Results. In patients, receiving cabozantinib as the first-line therapy, objective response rate was 66.7 %, tumor control was reached in 100 % of cases. Median time to the objective response was 2.6 (1.9-3.6) months, median objective response duration - 13.2 (6.2-21.5) months. Median progression-free survival (PFS) and overall survival (OS) were not reached, 6- and 12-months PFS was 77.8 % and 77.8 %, 6- and 12-months OS - 88.9 % and 88.9 % respectively. Cabozantinib as the second and subsequent lines of therapy provided objective response rate of 34.9 %, tumor control rate - 97.6     %. Median time to the objective response was 2.5 (1.8-4.1) months, median objective response duration - 12.6 (5.5-27.3) months. Median PFS was not reached (6- and 12-months PFS - 92.5 % and 73.1 % respectively), median OS was 32.6      months (6- and 12-months OS - 97.4 % and 80.8 % respectively). Any adverse events (AE) developed in 88.8 %, AE grade III-IV - in 32.6 % of cases. Most frequent AE grade III-IV included arterial hypertension (18.5 %), diarrhea (6.5 %) and palmar-plantar erythrodysesthesia (6.5 %). Unacceptable toxicity demanded treatment cancellation in 2.2 %, therapy interruption - in 16.3 % and dose reduction - in 30.4 % of patients.Conclusion. Cabozantinib as the first and subsequent lines of therapy for metastatic renal cell carcinoma patients in the real world practice demonstrated high efficacy and better tolerability comparing with population assigned for cabozantinib monotherapy in the randomized phase II-III trials.Цель исследования - оценка эффективности и безопасности кабозантиниба у неотобранных пациентов с метастатическим почечно-клеточным раком в 1-й и последующих линиях терапии.Материалы и методы. В многоцентровое обсервационное исследование было последовательно включено 92 пациента с морфологически верифицированным метастатическим почечно-клеточным раком, получавшие кабозантиниб (60 мг/сут) в 16 российских центрах. Медиана возраста больных составила 56 (19-79) лет, соотношение мужчин и женщин - 3:1. На момент старта терапии кабозантинибом ECOG PS 2 имел место у 27,2 % пациентов. Наиболее частым гистологическим вариантом рака почки являлся светлоклеточный (90,2 %). У большинства больных диагностированы синхронные (71,7 %) множественные метастазы (60,9 %). Предшествующая нефрэктомия выполнена в 87,0 % случаев. Прогноз по шкале International Metastatic Renal Cancer Database Consortium (IMDC) был расценен как благоприятный у 5,4 %, промежуточный - у 58,7 % и неблагоприятный - у 35,9 % больных. Кабозантиниб в качестве терапии 1-й линии применялся в 9 (9,8 %), после 1-5 линий системной терапии - в 83 (90,2 %) случаях. Медиана наблюдения за всеми пациентами составила 11 (2,3-44,5) мес.Результаты. У больных, получавших кабозантиниб в качестве терапии 1-й линии, частота объективного ответа составила 66,7 %, контроль над опухолью достигнут в 100 % случаев. Медиана времени до объективного ответа равнялась 2,6 (1,9-3,6) мес, медиана продолжительности объективного ответа - 13,2 (6,2-21,5) мес. Медианы беспрогрессивной выживаемости (БПВ) и общей выживаемости (ОВ) не достигнуты, 6- и 12-месячная БПВ составила 77,8 и 77,8 %, 6- и 12-месячная ОВ - 88,9 и 88,9 % соответственно. Кабозантиниб в качестве 2-й и последующих линий терапии обеспечил частоту объективного ответа 34,9 % и частоту контроля над опухолью, достигшую 97,6 %. Медиана времени до объективного ответа составила 2,5 (1,8-4,1) мес, медиана продолжительности объективного ответа - 12,6 (5,5-27,3) мес. Медиана БПВ не достигнута (6- и 12-месячная БПВ - 92,5 и 73,1 % соответственно), медиана ОВ составила 32,6 мес (6- и 12-месячная ОВ - 97,4 и 80,8 % соответственно). Любые нежелательные явления развились в 88,8 %, нежелательные явления III-IV степени - в 32,6 % случаев. Наиболее частыми нежелательными явлениями III-IV степени были артериальная гипертензия (18,5 %), диарея (6,5 %) и ладонно-подошвенная эритродизестезия (6,5 %). Неприемлемая токсичность потребовала отмены лечения у 2,2 %, прерывания терапии - у 16,3 % и снижения дозы - у 30,4 % больных.Заключение. Кабозантиниб в 1-й и последующих линиях терапии метастатического почечно-клеточного рака в реальной мировой практике продемонстрировал высокую эффективность и лучшую переносимость по сравнению с результатами рандомизированных исследований II-III фаз

COMBINED METHOD OF SURGICAL TREATMENT FOR PARALYTIC LAGOPHTHALMOS WITH POLYTETRAFLUOROETHYLENE (PTFE) IMPLANTS

The purpose is to develop a combined method of paralytic lagophthalmos treatment using polytetrafluoroethylene (PTFE) implants. Material and methods. The clinical group consisted of 20 patients (8 males, 12 females) aged 31-69 (44.7 ± 5.3 years) with facial nerve paralysis, who were treated within the last 3 years. The onset of paralytic lagophthalmos took place 1-9 years (3.3 ± 2.1 years) before surgical treatment. A typical clinical picture was as follows: lagophthalmos 9-16 mm (13.3 ± 3.1 mm), retraction of the upper eyelid 3-6 mm (4.1 ± 1.1 mm), atony and ectropion of the lower eyelid, epitheliopathy (11 cases), leucoma (9 cases). Visual acuity was 0.01-1.0 (0.3 ± 0.15). All patients had surgery for lagophthalmos with the use of a PTFE implant. In 5 patients, surgery was performed in two stages (separately for the upper and the lower eyelids), in 15 cases it was performed simultaneously, using a combined method The implant was a porous PTFE tape 200-300 μm thick, either solid or with perforations. The follow-up period was 6 months to 3 years (1.9 ± 0.9 years). Results. No complications have been revealed either in the early or late follow-up periods. All patients had positive dynamics of the following parameters: the size of lagophthalmos, the presence of ectropion of the lower eyelid, retraction of the upper eyelid, the condition of the cornea. The evaluation was conducted 6 months after surgery or later. In all cases, lagophthalmos decreased significantly, the residual lagophthalmos was 2-5 mm (3.1 ± 1.3 mm). The condition of the cornea in patients with keratopathy improved, vision increased to reach 0.1 1.0 (0.5 ± 0.25). Conclusion. In the proposed method, the lagophthalmos is eliminated by lower eyelid strengthening and upper eyelid recession. PTFE implants provide a stable effect. For citation: Filatova I.A., Shemetov S.A. Combined method of surgical treatment for paralytic lagophthalmos with polytetrafluoroethylene (PTFE) implants. Russian ophthalmological journal. 2018; 11 (2): 27-34. doi: 10.21516/2072-0076-2018-11-2-27-34 (In Russian)

Modified eyeball evisceration surgery using radio wave surgery technique

Purpose: development of a modified eye evisceration technique using radio wave surgery and evaluation of its effectiveness. Material and methods. The results of treatment of 383 patients were analyzed. The majority were men aged 19 to 55 (62.7 %) after eye removal. Of the treated patients, 328 (85.6 %) were operated due to the trauma. Surgery techniques included enucleation with stump plastics (131 patients, comparison group I), evisceration with sclera posterior pole resection, neurectomy and stump plastics (83 patients, comparison group II) and a modified method involving radio wave surgery and locomotor stump formation (169 patients, main group). For the orbital implant, carbon felt (Carbotextim) or Polytetrafluoroethylene (Teflon) were used. The patients were followed up for 6 months to 3 years. Results. In all cases, the results of surgical treatment and subsequent eye prosthetics were positive: their were traced in 344 patients (89.7 %). The best functional and cosmetic results assessed according to several features (retraction of the prosthesis and the upper lid into the orbit, stump and prosthesis mobility) were achieved after evisceration with stump plastics (in the main group and comparison group II).Conclusion. Using the above technique in subatrophy and enhancement of safety of the modified technique in posttraumatic uveitis validate the extension of indication to evisceration. For citations: Filatova I.A., Mohammad I.M., Shemetov S.A. Modified eyeball evisceration surgery using radio wave surgery technique. Russian ophthalmological journal. 2017; 10 (3): 84 92. doi: 10.21516/2072-0076-2017-10-3-84-92 (in Russian)

A modified method of evisceration of a subatrophic eyeball

The purpose is to develop a new method of evisceration of the eyeball in II–III degree subatrophies and assess its effectiveness. Material and methods. 172 patients with blind or incurable eyes who underwent one of the three operations: eye evisceration according to the newly developed technique involving an implantation of polytetrafluoroethylene grafts (main group), eyeball enucleation (comparison group I) and classical eye evisceration (comparison group II) were observed. Results. No complications were revealed either in the early or in the late period. A positive result was noted in all patients of the main group: average stump mobility was 140.1° ± 3.7° and average prosthesis mobility was 111.2° ± 4.0°. Prosthesis retraction after evisceration occurred 13.9 % less often than after enucleation (comparison group I), while the prosthesis retraction depth (mm) was 54.5 % lower than after enucleation. The retraction of the upper eyelid was also 26 % less common and its depth (mm) was 39.3 % lower