The effect of paired associative stimulation with a high-intensity cortical component and a high-frequency peripheral component on heart rate and heart rate variability in healthy subjects.

Objective: A novel protocol for paired associative stimulation (PAS), called high PAS, consists of high-intensity transcranial magnetic stimulation (TMS) and high-frequency peripheral nerve stimulation (PNS). High PAS was developed for spinal cord injury rehabilitation and targets plastic changes in stimulated pathways in the corticospinal tract, which improves motor function. As therapy interventions can last many weeks, it is important to fully understand the effects of high PAS, including its effect on the cardiovascular system. Heart rate variability (HRV) has been used to measure changes in both sympathetic and parasympathetic systems. Methods: We used short-term HRV measurements to evaluate the effects of one 20-min session of high PAS on 17 healthy individuals. HRV was recorded for 5 min before (PRE), during (STIM), immediately after (POST), 30 min after (POST30), and 60 min after (POST60) the stimulation. Five participants repeated the HRV setup with sham stimulation. Results: A significant decrease in low-frequency (LF) power (n.u.) (p = 0.002), low-frequency to high-frequency (HF) ratio (p = 0.017), in Poincaré plot [the standard deviation of RR intervals perpendicular to (SD1) and along (SD2) the line of identity SD2/SD1 ratio p < 0.001], and an increase in HF power (n.u.) (p = 0.002) were observed between PRE and STIM conditions; these changes were fully reversible immediately after stimulation. PRE to POST by 3% (p = 0.015) and continued to decline until POST60 by 5% (p = 0.011). LF power (ms2) (p = 0.017) and SD2 (p = 0.015) decreased from PRE to STIM and increased from PRE to POST (p = 0.025 and p = 0.017, respectively). The results from sham PAS exhibited a trend similar to active high-PAS stimulation. Conclusions: High PAS does not have sustained effects during 60-min follow-up on cardiovascular functions, as measured by HRV. None of the short-term results indicates activation of the sympathetic nervous system in healthy individuals. Observed changes in HRV indicate higher parasympathetic activity during stimulation, which is reversible, and is plausibly explained by the fact that the participants spend 20 min without moving, talking, or using phones while being stimulated.

Synergistic Effect of the Combined Action of Targeted and Photodynamic Therapy on HER2-Positive Breast Cancer.

Development of combined schemes for the treatment of oncological diseases is a promising strategy to improve the effectiveness of antitumor therapy. This paper shows the fundamental possibility of multiplying the antitumor effect by combining targeted and photodynamic therapy. It was demonstrated that sequential treatment of HER-2 positive breast cancer cells with the targeted toxin DARPin-LoPE and the photoactive compound photodithazine leads to a synergistic enhancement of their effect. In the future, this approach is intended to achieve the maximum therapeutic effect while minimizing the risks of negative side effects.

[THE EFFECTIVENESS OF TREATMENT IN CHILDREN WITH RIGID FLATFEET AND DYSFUNCTION OF THE POSTERIOR TIBIAL TENDON].

The rigid form of longitudinal flatfeet is one of the most common deformities of the foot in children. Dysfunction of the posterior tibial tendon is one of the causes of rigid foot formation with the development of pain syndrome, degenerative-dystrophic changes in the tarsal bones and joints. In most of these cases, conservative treatment is ineffective, prompting the use of surgical methods of foot correction - subtalar arthroereisis. However, currently, the indications for surgical treatment depending on the stage of posterior tibialis tendon dysfunction are poorly studied. The aim of the study was to analyze the clinical and radiological results of treatment of children with rigid flatfeet in the presence of posterior tibialis tendon dysfunction and to develop an algorithm of treatment depending on the stage of dysfunction. The results of treatment of 20 patients (40 cases) with the rigid form of longitudinal flatfeet during 12 months of follow-up were analyzed. In 18 patients (36 cases/90%) degenerative-dystrophic changes of the posterior tibial tendon according to Johnson and Strom classification were diagnosed. Conservative treatment with traditional methods was performed for 12 patients (24 cases), and 8 patients (16 cases) were operated on using subtalar arthroereisis. To analyze the results of treatment, clinical and radiological methods of investigation were performed. The use of conservative methods of treatment made it possible to obtain a positive result in patients with rigid flatfeet of the І severity degree and the absence of posterior tibialis tendon dysfunction. Subtalar arthroereisis was effective in children with II severity degree of rigid flatfeet and stage I of tendon dysfunction. Conservative treatment of rigid flatfeet is effective in the absence of posterior tibialis tendon dysfunction, and the use of subtalar arthroereisis is advisable in stage I of tendon dysfunction.

Optimization of Spondylosynthesis for Certain Thoracolumbar Burst Fractures.

Intermediate transpedicular fixation, i.e. additional insertion of transpedicular screws into the injured vertebrae, is an improvement to the most popular surgical intervention for spinal injuries, currently gaining widespread use in clinical practice. Unilateral insertion of transpedicular screws into the injured vertebrae allows combining the advantages of intermediate transpedicular fixation with the possibility to perform anterior column support without remounting the transpedicular system. The aim of the study was to use biomechanical computer modeling for evaluating the stability of intermediate transpedicular fixation components, which allow performing anterior column support if necessary. MATERIALS AND METHODS: DICOM files obtained during CT scan of a patient with intermediate thoracolumbar spine injury and the ANSYS software were used. Stability of the transpedicular system and supportability of the complementary Mesh implant installed with unilateral intermediate transpedicular screws were evaluated using computer modeling based on the finite element method. RESULTS: The values of stress and displacement fields for spine-hardware systems with various arrangements have been obtained. The maximum loads exceeding bone tissue strength (153-161 MPa) were registered for standard 4-screw system (190 MPa) when modeling the load equivalent for walking and falling from a standing position. The use of the proposed fixation system arrangement supplemented with intermediate screws allows obtaining loads in the spine-hardware system not exceeding these thresholds. Complementary eccentric Mesh implant enhances fixation stability of the transpedicular system with intermediate screws. CONCLUSION: The results show the high degree of mechanical stability of the proposed hardware arrangement and its potential efficacy for thoracolumbar transitional vertebra stabilization.

Treatment Tactics for Patients with Isolated Injuries of the Fifth Lumbar Vertebra.

The major management technique for lumbar burst fractures is transpedicular fixation (TPF). However, in relation to fractures of the L5 vertebra, this tactic often has no advantages over conservative treatment, and, therefore, it is expected to be supplemented with anterior decompression and reconstruction of the anterior column of the L5 vertebra. The aim of the study was to determine the most optimal treatment tactics for patients with isolated burst fractures of the fifth lumbar vertebra. Materials and Methods: We performed a retrospective study of 58 patients treated for isolated burst L5 fractures. 12 patients refused to undergo surgery and received conservative outpatient treatment. TPF was performed in 27 patients; circular spondylosynthesis (TPF + anterior column support with a Mesh implant) - in 19 patients. The effectiveness of the treatment was assessed by clinical and introscopic research methods. Results: The radiological and functional outcomes of surgery with conventional TPF for isolated L5 burst fractures are generally comparable with the outcomes of conservative treatment. In 26% of the patients, the instability of the metal construction developed within 12 months after surgical intervention. Supplementing the transpedicular system with wedging anterior column support with a Mesh implant ensures preservation in 21%, and improves the parameters of the sagittal profile of the lumbosacral transition in 79% of cases.

Enabling and promoting walking rehabilitation by paired associative stimulation after incomplete paraplegia: a case report.

INTRODUCTION: Paired associative stimulation (PAS) is a combination of transcranial magnetic stimulation (TMS) and peripheral nerve stimulation (PNS) and induces plastic changes in the human corticospinal tract. We have previously shown that PAS consisting of TMS pulses given at 100% of stimulator output and high-frequency PNS is beneficial for motor rehabilitation of patients with a chronic incomplete spinal cord injury (SCI). The therapeutic possibilities of this PAS variant for walking rehabilitation of paraplegic patients are unexplored. CASE PRESENTATION: A 47-year old man with traumatic incomplete paraplegia (AIS D, neurological level T7) received PAS to his left leg for 3 months at 12 months post injury (PAS1) and for an additional 3 months at 24 months post injury (PAS2). The right leg had normal AIS scores and was not stimulated. Before PAS, the patient was nonambulatory, could not stand without weight support, and was consequently not eligible for conventional walking rehabilitation. After PAS1, the patient could stand for 1.5 min and take 13 steps (24 steps in follow up) on parallel bars without weight support and was enrolled into conventional walking rehabilitation. He achieved independent walking ability with a rollator. During PAS2, walking distance increased 2.4 times faster than during the preceding year. The left leg AIS score and spinal cord independence measure mobility subscore increased. No adverse effects were detected. DISCUSSION: This is the first report of PAS with a high-frequency peripheral component that enabled and promoted walking rehabilitation. Together with previous reports on this technique, this result encourages further research into its therapeutic potential and mechanism.

Multifunctional Complexes Based on Photoluminescent Upconversion Nanoparticles for Theranostics of the HER2-Positive Tumors.

Combining diagnostic and therapeutic functions in one agent is a promising strategy in the development of personalized approaches to the treatment of cancer. The opportunity to combine diagnostics and therapy appeared with the development of nanobiotechnologies and was realized in the concept of theranostics. To date, a number of promising agents based on nanomaterials capable of diagnosing, targeted therapeutic effects, and monitoring the response of tumor cells were obtained within the approach of theranostics. In this work, a new type of theranostic complexes based on upconversion nanoparticles coated with polyethylene glycol and functionalized with the DARPin-LoPE recombinant targeted toxin was developed. Selective binding of complexes to human breast adenocarcinoma cells overexpressing the HER2 receptor and specific toxicity to them were shown.

Restoration of hand function with long-term paired associative stimulation after chronic incomplete tetraplegia: a case study.

Introduction: This case study explores the gains in hand function in an individual with a chronic spinal cord injury (SCI). The intervention was long-term paired associative simulation (PAS). We aimed to provide PAS until full recovery of hand muscle strength occurred, or until improvements ceased. Case presentation: A 46-year-old man with traumatic C7 AIS B tetraplegia was administered PAS three times per week. After 24 weeks, PAS was combined with concomitant motor training of the remaining weak hand muscles. Outcome measures included the manual muscle test (MMT), motor-evoked potentials (MEPs), F-responses, hand functional tests, and the spinal cord independence measure (SCIM). Discussion: After 47 weeks of PAS the subject had improved self-care and indoor mobility and was able to perform complex motor tasks (SCIM score improved from 40 to 56). His left hand regained maximum MMT score (total 75; increase of score from baseline condition 19); the effect remained stable in the 32-week follow up. In the right-hand muscles, MMT scores of 4-5 were observed in follow up (total 71; increase from baseline 48). Improved values were also observed in other outcomes. This is the first demonstration of long-term PAS restoring muscle strength corresponding to MMT scores of 4-5 in an individual with chronic SCI. The effect persisted for several months, indicating that PAS induces stable plastic changes in the corticospinal pathway.

Metalloproteinase inhibitor GM6001 delays regeneration in holothurians.

The effect of the GM6001 metalloproteinase inhibitor on the regeneration of ambulacral structures in Eupentacta fraudatrix has been investigated. Inhibition of proteinase activity exerts a marked effect on regeneration, being dependent on the time when GM6001 is injected. When administration of the inhibitor begins on day 3 post-injury, regeneration is completely abolished, and the animals die. This means that early activation of proteinases is crucial for triggering the regenerative process in holothurians. When GM6001 in first injected on day 7 post-injury, the regeneration rate decreases. However, this effect has proven to be reversible: when inhibition ceases, the regeneration resumes. The effect of the inhibitor is manifested as a retarded degradation of the extracellular matrix, the lack of cell dedifferentiation, and, probably, a slower cell migration. The gelatinase activity is detected in all the regenerating organs of E. fraudatrix. In the holothurian Cucumaria japonica, which is not capable of healing skin wounds and ambulacrum reparation, no gelatinase activity was observed at the site of damage. A suggestion is made that proteinases play an important role in regeneration in holothurians. The most probable morphogenesis regulators are matrix metalloproteinases with gelatinase activity.

[Indicators of kidney damage in patients with multiple myeloma in the process of chemotherapeutic treatment.]

The purpose was to study the level of acute kidney injury markers cystatin C, KIM-1, IL-18, NGAL and L-FABR in the blood and urine of patients with the initially identified secreting multiple myeloma (MM) before and during chemotherapeutic treatment. The content of renal markers was examined by ELISA using commercial kits. The study included 23 patients with MM who received 6-8 21-day cycles of chemotherapy (CT) according to the VCD scheme. The results were compared in the main group of 13 patients who had a selective plasma exchange a day before each of the cycles of HT with the use of the Evaclio plasma separator and in a control group of 10 patients treated without extracorporeal detoxification. MM patients before treatment showed an increase in blood IL-18 level of 8.6 times, KIM-1 - 3.1 times, L-FABR - 57.4%, cystatin C - 48.4% and also a decrease in the level of NGAL in 75% of patients by 74.3% compared to the level in healthy, while in the urine initially increased only KIM-1 content by 2.4 times and NGAL by 2.6 times. Conducting multi-course chemotherapy with previous plasma exchange had a more lenient effect on MM patients, as evidenced by a lower KIM-1 level in blood and urine after 1 and 2 courses of HT, as well as IL-18 in blood and urine after 1 course of HT in patients of primary group compared with the control group. For patients with a fatal outcome, a sharp increase in the levels of cystatin C, NGAL and L-FABR is characteristic. The analysis of the dynamics of the studied markers of renal damage indicates the correlation of their level with the clinical features of individual patients, the success and tolerability of chemotherapeutic treatment of MM.

Stably Fluorescent Cell Line of Human Ovarian Epithelial Cancer Cells SK-OV-3ip-red.

Stable red fluorescing line of human ovarian epithelial cancer cells SK-OV-3ip-red was generated expressing gene coding for protein TurboFP635 (Katushka) fluorescing in the far-red spectrum region with excitation and emission peaks at 588 and 635 nm, respectively. Fluorescence of SK-OV-3ip-red line remained high during long-term cell culturing and after cryogenic freezing. The obtained cell line SK-OV-3ip-red can serve a basis for a model of a scattered tumor with numerous/extended metastases and used both for testing anticancer drugs inhibiting metastasis growth and for non-invasive monitoring of the growth dynamics with high precision.

Interplay between thyroxin, BDNF and GABA in injured neurons.

Accumulating experimental evidence suggests that groups of neurons in the CNS might react to pathological insults by activating developmental-like programs for survival, regeneration and re-establishment of lost connections. For instance, in cell and animal models it was shown that after trauma mature central neurons become dependent on brain-derived neurotrophic factor (BDNF) trophic support for survival. This event is preceded by a shift of postsynaptic GABAA receptor-mediated responses from hyperpolarization to developmental-like depolarization. These profound functional changes in GABAA receptor-mediated transmission and the requirement of injured neurons for BDNF trophic support are interdependent. Thyroid hormones (THs) play a crucial role in the development of the nervous system, having significant effects on dendritic branching, synaptogenesis and axonal growth to name a few. In the adult nervous system TH thyroxin has been shown to have a neuroprotective effect and to promote regeneration in experimental trauma models. Interestingly, after trauma there is a qualitative change in the regulatory effect of thyroxin on BDNF expression as well as on GABAergic transmission. In this review we provide an overview of the post-traumatic changes in these signaling systems and discuss the potential significance of their interactions for the development of novel therapeutic strategies.

Expression of G-protein coupled receptors in Escherichia coli for structural studies.

To elaborate a high-performance system for expression of genes of G-protein coupled receptors (GPCR), methods of direct and hybrid expression of 17 GPCR genes in Escherichia coli and selection of strains and bacteria cultivation conditions were investigated. It was established that expression of most of the target GPCR fused with the N-terminal fragment of OmpF or Mistic using media for autoinduction provides high output (up to 50 mg/liter).

Bacterial synthesis, purification, and solubilization of membrane protein KCNE3, a regulator of voltage-gated potassium channels.

An efficient method is described for production of membrane protein KCNE3 and its isotope labeled derivatives ((15)N-, (15)N-/13C-) in amounts sufficient for structural-functional investigations. The purified protein preparation within different detergent micelles was characterized using dynamic light scattering, CD spectroscopy, and NMR spectroscopy. It is shown that within DPC/LDAO micelles the protein is in monomeric form and acquires mainly alpha-helical conformation. The existence of cross-peaks for all glycines of the (15)N-HSQC NMR spectra as well as relatively small line widths (~20 Hz) confirm the high quality of the preparation and the possibility of obtaining structural-dynamic information on KCNE3 by high resolution heteronuclear NMR spectroscopy.

Rhamnolipid-biosurfactant permeabilizing effects on gram-positive and gram-negative bacterial strains.

The potential of biosurfactant PS to permeabilize bacterial cells of Pseudomonas aeruginosa, Escherichia coli, and Bacillus subtilis on growing (in vivo) and resting (in vitro) cells was studied. Biosurfactant was shown to have a neutral or detrimental effect on the growth of Gram-positive strains, and this was dependent on the surfactant concentration. The growth of Gram-negative strains was not influenced by the presence of biosurfactant in the media. Cell permeabilization with biosurfactant PS was shown to be more effective with B. subtilis resting cells than with Pseudomonas aeruginosa. Scanning-electron microscopy observations showed that the biosurfactant PS did not exert a disruptive action on resting cells such that it was detrimental to the effect on growing cells of B. subtilis. Low critical micelle concentrations, tender action on nongrowing cells, and neutral effects on the growth of microbial strains at low surfactant concentrations make biosurfactant PS a potential candidate for application in different industrial fields, in environmental bioremediation, and in biomedicine.

Nanoassemblies designed from semiconductor quantum dots and molecular arrays.

The formation of nanoassemblies of CdSe/ZnS quantum dots (QD) and pyridyl-substituted free-base porphyrin (H(2)P) molecules has been spectroscopically identified by static and time-resolved techniques. The formation of nanoassemblies has been engineered by controlling the type and geometry of the H(2)P molecules. Pyridyl functionalization gives rise to a strong complex formation accompanied by QD photoluminescence (PL) quenching. For some of the systems, this quenching is partly related to fluorescence resonance energy transfer (FRET) from the QD to H(2)P and can be explained according to the Förster model. The quantitative interpretation of PL quenching due to complexation reveals that (i) on average only about (1)/(5) of the H(2)P molecules at a given H(2)P/QD molar ratio are assembled on the QD and (ii) only a limited number of "vacancies" accessible for H(2)P attachment exist on the QD surface.

Biosurfactant-rhamnolipid effects on yeast cells.

AIM: The aim of this work was to study the effect of the novel surfactant PS from Pseudomonas sp. S-17 on Saccharomyces cerevisiae 83-20 yeast cells and to compare it with the effect of the well known surfactant Triton X-100. METHODS AND RESULTS: The effect of surfactants was investigated on the cells during growth, and on the separated cells. The cell-permeabilizing effect of surfactants was studied by following the release of protein and some enzyme activities. The biosurfactant did not affect the culture growth kinetics, and altered the polypeptide profiles of cells and membrane proteins in the same way as Triton X-100. CONCLUSION: Results of this study demonstrate that biosurfactant PS and Triton X-100 have a similar type of action, mainly surface located, and that they do not affect the intracellular structures of yeast cells. SIGNIFICANCE AND IMPACT OF THE STUDY: A novel surfactant PS was isolated from Pseudomonas sp. S-17. A mild effect of PS on yeast cells was demonstrated. The results indicate the ecological safety of the biosurfactant and its potential use in the development of environmentally-benign and efficient cleaning technologies.

Three-dimensional structure of binase in solution.

We present the spatial structure of binase, a small extracellular ribonuclease, derived from 1H-NMR* data in aqueous solution. The total of 20 structures were obtained via torsion angle dynamics using DYANA program with experimental NOE and hydrogen bond distance constraints and phi and chi1 dihedral angle constraints. The final structures were energy minimised with ECEPP/2 potential in FANTOM program. Binase consists of three alpha-helices in N-terminal part (residues 6-16, 26-32 and 41-44), five-stranded antiparallel beta-sheet in C-terminal part (residues 51-55, 70-75, 86-90, 94-99 and 104-108) and two-stranded parallel beta-sheet (residues 22-24 and 49-51). Three loops (residues 36-39, 56-67 and 76-83), which play significant role in biological functioning of binase, are flexible in solution. The differences between binase and barnase spatial structures in solution explain the differences in thermostability of binase, barnase and their hybrids.

[Activation of L-amino acids by aminoacyl-tRNA-synthetases from yeast Candida utilis IBPM-405]. / Aktivatsiia l-aminokislot preparatom aminoatsil-tRNK-sintetaz iz drozhzhei candida utilis ibfm-405

The procedure for isolating aminoacyl-tRNA-synthetases from yeast Candida utilis IBPM-405 was developed. The rate of activation of L-amino acids in the formation of hydroxamates was different. Aspartic acid, asparagine, glutamic acid, tryptophane, phenyl alanine and methionine underwent the highest activation. The activation of alanine, arginine, hydroxyproline, serine and isoleucine was insignificant. Using aspartic acid, it was shown that the hydroxamate formation was ATP-stimulated and that the amount of hydroxamate increased with a rise of the protein concentration in the mixture to 9-10 mg/ml. The hydroxamate formation was inhibited by p-chloromercury-benzoate and heavy metal ions. Yeast aminoacyl-tRNA-synthetases showed L-aspartic and L-glutamic activities that were independent from Mg++ ions and ATP.