Copper-Catalyzed Three-Component Carboiminolactonization of Electron-Deficient Olefins.

Herein we report the three-component copper-catalyzed carboiminolactonization of α,ß-unsaturated carbonyl derivatives. In the presence of a Cu(I) catalyst, α-haloesters, electron-deficient alkenes, and primary amines couple to generate γ-iminolactones in a single step. The scope of the reaction is explored with respect to the three coupling partners. Nineteen examples are presented with yields of these hydrolytically labile heterocycles of up to 69%. Mechanistic investigations support the formation of an oxocarbenium by way of an atom transfer radical addition (ATRA) intermediate.

Cu-Catalyzed Three-Component Carboamination of Electron Deficient Olefins.

The copper-catalyzed three-component carboamination of atropates for the synthesis of α-aryl amino acid derivatives is presented. The scope of the reaction is explored with respect to all three coupling partners: the alkyl halide, the atropate, and the aryl amine. A total of 41 examples are included, with yields of ≤92%. Both primary and secondary aryl amines participate in the carboamination along with α-haloesters, nitriles, and perfluoroiodoalkanes. Mechanistic investigations support a radical mechanism involving Cu-mediated C-N bond formation with the radical adduct.

Mineral water inhalations for bronchial asthma: a meta-analysis.

Bronchial asthma (BA) is a common disease that contributes significantly to the incidence rate and death rate worldwide. A widespread treatment method is the use of inhalations of mineral waters, with conflicting information about their effectiveness. Purpose of the study was to assess the generalized effect power of the course of inhalations of mineral waters on the disease progress in patients with BA. A search of randomized clinical studies in data bases Pubmed, EMBASE, ELibrary, MedPilot amd CyberLeninka, according to PRISMA strategy, published between 1986 and July 2021. Standardized difference of mean values and their 95% of CI were employed for calculation using the random effects model. The meta-analysis drawing on 1266 sources included 14 studies, with 2 of them being randomized controlled clinical studies, including the results of the treatment of 525 patients. All 14 articles contain a conclusion that the inhalation of mineral water has a positive effect on the course of the disease in patients with BA. The analysis demonstrated that the group of patients after mineral water inhalations, compared with the control group, showed improvement of forced expiratory volume (FEV1), expressed both in % of the norm and in liters. The standardized difference of mean values FEV1 (%) (Hedge's g) was 8.2 (95% CI: 5.87 - 10.59; 100%), FEV1 values (liter.) (Hedge's g) was 0.69 (95% CI: -0.33-1.05). A significant heterogeneity of the results of individual studies was found (Q=124.96; tau2 = 14.55, I2 = 69.13%, p<0.0001 and Q=2.35; tau2 = 0, I2 = 0%, p<0.0001). Patients with mild, moderate, and hormone-dependent BA with a controlled and partially controlled disease course, after mineral water inhalations, compared with the control group, demonstrated a statistically significant decrease in the frequency and intensity of the cardinal symptoms of BA and improvement of FEV1.

Postmyocardial infarction ventricular septal defect and ventricular aneurysm repair with a "double-patch frame" technique.

BACKGROUND: Postmyocardial infarction intraventricular septal rupture is a life-threatening medical condition. Surgical management of postmyocardial infarction ventricular septal defects (PIVSDs) is associated with a 60% mortality and a 40% incidence of residual ventricular septal defects (rVSDs). Our study aimed to describe our modification of the "Double-patch" technique of PIVSD repair without using a biological glue and present its postoperative complications and survival. METHODS: This was a retrospective observational study. The Bakoulev's Scientific Center of Cardiac Surgery patient admission and discharge database was reviewed from March 2002 to April 2021. The inclusion criterion was PIVSD. Exclusion criteria were conservative treatment, transcatheter closure of PIVSD, PIVSD closure with an interventricular septum patch, and chronic PIVSDs. The study outcomes were echocardiographic parameters of cardiac function, postoperative complications, and mortality. RESULTS: Forty nine patients met the study eligibility criteria. Comparison of echocardiographic data of cardiac function demonstrated reduction in the postoperative period end-diastolic (201.4 ± 59.6 ml vs. 118 [range: 76-207] ml; p < .0005) and end-systolic volumes (106 [51-208] ml vs. 66 [40-147] ml; p < .0005). One (2%) patient developed hemodynamically significant rVSD that required the second run of cardiopulmonary bypass and rVSD closure. Thirteen (26.5%) patients died in the hospital. The overall mortality rate for the study period was 11.4/100 person-years (95% confidence interval [CI]: 6.9-19.0/100 person-years). In these patients, 1-year survival was 68.2% (95% CI: 52.3%-79.8%) and 5-year survival was 63.1% (95% CI: 45.1%-76.7%). CONCLUSION: The "Double-patch frame" technique restores LV dimensions, has a low rate of hemodynamically significant rVSDs and mortality.

Intradermal injection of lidocaine with a microneedle device to provide rapid local anaesthesia for peripheral intravenous cannulation: A randomised open-label placebo-controlled clinical trial.

BACKGROUND: Peripheral venous cannulation is one of the most common procedures in medicine. It is associated with noticeable pain and apprehension, although in most cases it is performed without any anesthesia due to lack of a painless, cost-effective option, which would provide rapid local anesthesia with subsequent significant reduction in the experienced pain. We conducted an open-label placebo-controlled clinical trial to evaluate the efficacy and safety of a 2% lidocaine injection using the commercially available microneedle device MinronJet600 (NanoPass Technologies Ltd, Israel) to achieve rapid local anesthesia prior to peripheral venous cannulation. METHODS: One hundred and two subjects were randomly allocated into two groups. In the first group, 100µL of lidocaine hydrochloride (2%) was injected intradermally to subjects using the MicronJet600 device in the left arm (MJ-Lido) and 100µL of saline was injected intradermally using the device in the right arm (MJ-Saline). In the second group, 100µL of lidocaine hydrochloride (2%) was injected using the MicronJet600 device into the left arm (MJ-Lido), with no injection into the right arm of subjects (No pretreatment). In both groups the intradermal injection was performed at the cannulation site prior to insertion of a 18G cannula into a median cubital vein in both arms. As a primary variable, a score of cannulation-induced pain was indicated by subjects using a 100-point visual analog scale immediately after cannulation. As a secondary variable, subjects in Group 2 also indicated their preference to receive the anaesthetic injection with MicronJet600 in the future by using the 5-point Likert scale. Also, as a secondary variable, the duration of skin numbness after lidocaine injection was indicated by performing a superficial pin-prick with a 27G needle at 15, 30 and 45 minutes, at distances of 1, 2 and 3 centimeters from the injection site. RESULTS: A significant pain reduction (11.0-fold) was achieved due to the lidocaine injection compared to the cannulation without any pretreatment (p< 0.005). After the lidocaine injection the anesthesia was effective up to 2 centimeters from the injection site and remained for up to 30 minutes. Eighty percent of subjects from the second group preferred cannulation after the lidocaine injection over cannulation without any pretreatment. No significant side effects were identified. CONCLUSION: Intradermal injection of anaesthetic with Micronjet600 was found to be a safe and effective option for providing rapid local anesthesia for peripheral intravenous cannulation. TRIAL REGIATRATION: The clinical trial was registered, before the patient enrollment began, in the Research Registry publicly accessible database (registration identifier: researchregistry4662). Also, the trial was registered in ClinicalTrials.gov (registration identifier: NCT05108714) after its completion.

Multisource Feedback in the Trauma Context: Priorities and Perspectives.

OBJECTIVES: Trauma resuscitations require competence in both clinical and nonclinical skills but these can be difficult to observe and assess. Multisource feedback (MSF) is workplace-based, involves the direct observation of learners, and can provide feedback on nonclinical skills. We sought to compare and contrast the priorities of multidisciplinary trauma team members when assessing resident trauma team captain (TTC) performance. Additionally, we aimed to describe the nature of the assessment and perceived the utility of incorporating MSF into the trauma context. METHODS: A convenience sample of 10 trauma team activations were observed. Following each activation, the attending physician trauma team leader (TTL), TTC, and a registered nurse (RN) participated in a semistructured interview. MSF was not provided to the TTC for the purpose of this study because MSF was not part of the assessment process of TTCs at the time of this study and maintaining anonymity may have encouraged more honest interview responses. Transcripts from each assessor group (TTL, TTC, RN) were coded and assigned to one of the five crisis resource management skills: leadership, communication, situational awareness, resource utilization, and problem-solving. Comments were also coded as positive, negative, or neutral as interpreted by the coder. RESULTS: All assessor groups mentioned communication skills most frequently. After communication, the RN and TTC groups commented on situational awareness most frequently, comprising 15 and 29% of their total responses, respectively, whereas 31% of the TTL comments focused on leadership skills. The RN and TTL groups provided positive assessments, with 51 and 42% of their respective comments coded as positive. Forty-five percent of self-assessment comments in the TTC group were negative. All (100%) of the TTC and TTL respondents felt that incorporating MSF would add to the quality of feedback, only 66% of the RN group felt that way. CONCLUSIONS: We found that each assessor group brings a unique focus and perspective to the assessment of resident TTC performance. The future inclusion of MSF in the trauma team context has the potential to enhance the learning environment in a clinical arena that is difficult to directly observe and assess.

Indigenous women's reproductive health in the Arctic zone of Western Siberia: challenges and solutions.

In the Russian Arctic, alarming trends (shortage of nomadic Indigenous women, high reproductive loss, child mortality rates) indicate long-term changes towards demographic decline. This study aimed at comparing some indicators of the reproductive health (childbirth rates, number of pregnancies, pregnancy loss) of Indigenous and non-Indigenous women in the exploration of cultural and social factors influencing reproductive behaviour. A multidisciplinary approach draws on methods of medicine, sociology and health economics. It includes data of the women's reproductive health collected from surveys of 879 women (of whom 627 were Indigenous) during expeditions to the settlements and the tundra of the Yamal-Nenets Autonomous Okrug in 2013-2019. In the tundra, 66.7% of registered Indigenous women's pregnancies resulted in childbirth, 7.8% in induced abortions, 25.5% in spontaneous miscarriage. More than three children were delivered by 59.1% of Indigenous women. Most Indigenous families suffered from high pregnancy loss. Marriage between close relatives was 27.0%. Child mortality equalled 24.4%, three times higher than in the population of the settlements and eight times higher than in families immigrating from other regions. The survival of Indigenous peoples significantly depends on women's reproductive health and sufficient medical service that requires targeted policy.

AG11, a novel dichloroflavanone derivative with anti-mitotic activity towards human bladder cancer cells.

BACKGROUND: New chemotherapy drugs should be investigated to improve survival of patients with advanced bladder cancer. Here, we report the synthesis and evaluation of AG11, a new flavanone derivative obtained through cyclization of its chalcone precursor CB11. MATERIALS AND METHODS: The effect of AG11 on cell viability was evaluated by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide assay and apoptotic cell death was analyzed by flow cytometry. Finally, the effect of AG11 on tubulin polymerization in vitro and microtubule distribution across the cells was investigated. RESULTS: AG11 was found to have an IC50 (half-maximal inhibitory concentration) of 4.6 µM and its inhibitory effect on RT4 cells proliferation is associated with a cell-cycle arrest in G2+M phases followed by apoptosis after a 48 h treatment. AG11 prevented polymerization of purified tubulin in a concentration-dependent manner in vitro and disrupted mitotic spindle formation in cells. CONCLUSION: AG11 appears to be an attractive scaffold for further development of a structurally simpler new anti-microtubule agents.

Effect of Peierls transition in armchair carbon nanotube on dynamical behaviour of encapsulated fullerene.

The changes of dynamical behaviour of a single fullerene molecule inside an armchair carbon nanotube caused by the structural Peierls transition in the nanotube are considered. The structures of the smallest C20 and Fe@C20 fullerenes are computed using the spin-polarized density functional theory. Significant changes of the barriers for motion along the nanotube axis and rotation of these fullerenes inside the (8,8) nanotube are found at the Peierls transition. It is shown that the coefficients of translational and rotational diffusions of these fullerenes inside the nanotube change by several orders of magnitude. The possibility of inverse orientational melting, i.e. with a decrease of temperature, for the systems under consideration is predicted.

The role of molecular clusters in the filling of carbon nanotubes.

We have demonstrated that the ability of fullerenes to form clusters is essential for the filling of single-walled carbon nanotubes in solution. In solutions where C60 exists in the form of discrete solvated molecules (e.g., in CS2) no fullerene encapsulation in nanotubes takes place, as the large molar excess of solvent compared to solute prohibits C60 from entering the nanotubes. However, in solutions containing large clusters of C60 (e.g., in n-hexane) nanotubes become densely filled with fullerene molecules despite the large excess of solvent. The interactions between carbon nanotubes and fullerene clusters provide an efficient transport of C60 into nanotubes that avoids the detrimental effects of the solvent molecules. This new mechanism provides the first rational explanation of experiments involving nanotube filling with guest-molecules in solution.

Xenopus meiotic microtubule-associated interactome.

In metazoan oocytes the assembly of a microtubule-based spindle depends on the activity of a large number of accessory non-tubulin proteins, many of which remain unknown. In this work we isolated the microtubule-bound proteins from Xenopus eggs. Using mass spectrometry we identified 318 proteins, only 43 of which are known to bind microtubules. To integrate our results, we compiled for the first time a network of the meiotic microtubule-related interactome. The map reveals numerous interactions between spindle microtubules and the newly identified non-tubulin spindle components and highlights proteins absent from the mitotic spindle proteome. To validate newly identified spindle components, we expressed as GFP-fusions nine proteins identified by us and for first time demonstrated that Mgc68500, Loc398535, Nif3l1bp1/THOC7, LSM14A/RAP55A, TSGA14/CEP41, Mgc80361 and Mgc81475 are associated with spindles in egg extracts or in somatic cells. Furthermore, we showed that transfection of HeLa cells with siRNAs, corresponding to the human orthologue of Mgc81475 dramatically perturbs spindle formation in HeLa cells. These results show that our approach to the identification of the Xenopus microtubule-associated proteome yielded bona fide factors with a role in spindle assembly.

Role of non-coding RNAs in neurodegeneration and stress response in Drosophila.

The inherent limitations of genetic analysis in humans and other mammals as well as striking conservation of most genes controlling nervous system functioning in flies and mammals made Drosophila an attractive model to investigate various aspects of brain diseases. Since RNA research has made great progress in recent years here we present an overview of studies demonstrating the role of various non-coding RNAs in neurodegeneration and stress response in Drosophila as a model organism. We put special emphasis on the role of non-coding micro RNAs, hsr-omega transcripts, and artificial small highly structured RNAs as triggers of neuropathology including aggregates formation, cognitive abnormalities and other symptoms. Cellular stress is a conspicuous feature of many neurodegenerative diseases and the production of specialized proteins protects the nerve cells against aggregates formation. Therefore, herein we describe some data implicating various classes of non-coding RNAs in stress response in Drosophila. All these findings highlight Drosophila as an important model system to investigate various brain diseases potentially mediated by some non-coding RNAs including polyglutamine diseases, Alzheimer's disease, Huntigton's disease, and many others.

Probing FtsZ and tubulin with C8-substituted GTP analogs reveals differences in their nucleotide binding sites.

The cytoskeletal proteins, FtsZ and tubulin, play a pivotal role in prokaryotic cell division and eukaryotic chromosome segregation, respectively. Selective inhibitors of the GTP-dependent polymerization of FtsZ could constitute a new class of antibiotics, while several inhibitors of tubulin are widely used in antiproliferative therapy. In this work, we set out to identify selective inhibitors of FtsZ based on the structure of its natural ligand, GTP. We found that GTP analogs with small hydrophobic substituents at C8 of the nucleobase efficiently inhibit FtsZ polymerization, whereas they have an opposite effect on the polymerization of tubulin. The inhibitory activity of the GTP analogs on FtsZ polymerization allowed us to crystallize FtsZ in complex with C8-morpholino-GTP, revealing the binding mode of a GTP derivative containing a nonmodified triphosphate chain.

Purification and mass spectrometry identification of microtubule-binding proteins from Xenopus egg extracts.

Microtubule-binding proteins are conveniently divided into two large groups: MAPs (microtubule-associated proteins), which can stabilize, anchor, and/or nucleate microtubules, and motors, which use the energy of ATP hydrolysis for a variety of functions, including microtubule network organization and cargo transportation along microtubules. Here, we describe the use of Taxol-stabilized microtubules for purification of MAPs, motors, and their complexes from Xenopus egg extracts. Isolated proteins are analysed using sodium dodecyl sulfate gel electrophoresis and identified by various mass spectrometry and database mining technologies. Found proteins can be grouped into three classes: (1) known MAPs and motors; (2) proteins previously reported as associated with the microtubule cytoskeleton, but without a clearly defined cytoskeletal function; (3) proteins not yet described as having microtubule localization. Sequence-similarity methods employed for protein identification allow efficient identification of MAPs and motors from species with yet unsequenced genomes.

Biocompatibility and applications of carbon nanotubes in medical nanorobots.

The set of nanoelectromechanical systems (NEMS) based on relative motion of carbon nanotubes walls is proposed for use in medical nanorobots. This set includes electromechanical nanothermometer, jet nanoengine, nanosyringe (the last can be used simultaneously as nanoprobe for individual biological molecules and drug nanodeliver). Principal schemes of these NEMS are considered. Operational characteristics of nanothermometer are analyzed. The possible methods of these NEMS actuation are considered. The present-day progress in nanotechnology techniques which are necessary for assembling of NEMS under consideration is discussed. Biocompatibility of carbon nanotubes is analyzed in connection with perspectives of their application in nanomedicine.

Pathogenic chaperone-like RNA induces congophilic aggregates and facilitates neurodegeneration in Drosophila.

Protein aggregation is a hallmark of many neurodegenerative diseases. RNA chaperones have been suggested to play a role in protein misfolding and aggregation. Noncoding, highly structured RNA recently has been demonstrated to facilitate transformation of recombinant and cellular prion protein into proteinase K-resistant, congophilic, insoluble aggregates and to generate cytotoxic oligomers in vitro. Transgenic Drosophila melanogaster strains were developed to express highly structured RNA under control of a heat shock promoter. Expression of a specific construct strongly perturbed fly behavior, caused significant decline in learning and memory retention of adult males, and was coincident with the formation of intracellular congophilic aggregates in the brain and other tissues of adult and larval stages. Additionally, neuronal cell pathology of adult flies was similar to that observed in human Parkinson's and Alzheimer's disease. This novel model demonstrates that expression of a specific highly structured RNA alone is sufficient to trigger neurodegeneration, possibly through chaperone-like facilitation of protein misfolding and aggregation.

Vaccinia-virus-induced cellular contractility facilitates the subcellular localization of the viral replication sites.

Poxviruses, such as vaccinia virus (VV), replicate their DNA in endoplasmic-reticulum-enclosed cytoplasmic sites. Here, we compare the dynamics of the VV replication sites with those of the attenuated strain, modified VV Ankara (MVA). By live-cell imaging, small, early replication sites of both viruses undergo motility typical of microtubule (MT)-motor-mediated movement. Over time, growing replication sites of VV collect around the nucleus in a MT-dependent fashion, whereas those of MVA remain mostly scattered in the cytoplasm. Surprisingly, blocking the dynein function does not impair the perinuclear accumulation of large VV replication sites. Live-cell imaging demonstrates that in contrast to small replication sites, large sites do not display MT-motor-mediated motility. Instead, VV infection induces cellular contractility that facilitates the collection of growing replication sites around the nucleus. In a subset of cells (30-40%), this VV-induced contractility is alternated by phases of directed cell migration, suggesting that the two processes may be linked. The MVA-infected cells do not display contractility or cell migration, supporting the idea that these cellular activities facilitate the efficient accumulation of the VV replication sites around the nucleus. We propose that the recently described cytoskeletal rearrangements induced by VV are a prerequisite for the observed cell contractility and migration activities that apparently contribute to the organization of the complex cytoplasmic life cycle of VV.

BENA435, a new cell-permeant photoactivated green fluorescent DNA probe.

N'-(2,8-Dimethoxy-12-methyl-dibenzo [c,h] [1,5] naphthyridin-6-yl)-N,N-dimethyl-propane-1,3-diamine (BENA435) is a new cell-membrane permeant DNA dye with absorption/emission maxima in complex with DNA at 435 and 484 nm. This new reagent is unrelated to known DNA dyes, and shows a distinct preference to bind double-stranded DNA over RNA. Hydrodynamic studies suggest that BENA435 intercalates between the opposite DNA strands. BENA435 fluoresces much stronger when bound to dA/dT rather than dG/dC homopolymers. We evaluated 14 related dibenzonaphthyridine derivatives and found BENA435 to be superior in its in vivo DNA-binding properties. Molecular modelling was used to develop a model of BENA435 intercalation between base pairs of a DNA helix. BENA435 fluorescence in the nuclei of cells increases upon illumination, suggesting photoactivation. BENA435 represents thus the first known cell-permeant photoactivated DNA-binding dye.

Homology-based functional proteomics by mass spectrometry: application to the Xenopus microtubule-associated proteome.

The application of functional proteomics to important model organisms with unsequenced genomes is restricted because of the limited ability to identify proteins by conventional mass spectrometry (MS) methods. Here we applied MS and sequence-similarity database searching strategies to characterize the Xenopus laevis microtubule-associated proteome. We identified over 40 unique, and many novel, microtubule-bound proteins, as well as two macromolecular protein complexes involved in protein translation. This finding was corroborated by electron microscopy showing the presence of ribosomes on spindles assembled from frog egg extracts. Taken together, these results suggest that protein translation occurs on the spindle during meiosis in the Xenopus oocyte. These findings were made possible due to the application of sequence-similarity methods, which extended mass spectrometric protein identification capabilities by 2-fold compared to conventional methods.