Introduction
The question of preoperative anxiety in an ophthalmological patient and its relief in an outpatient setting is becoming increasingly relevant due to the improvement of the technique and ...equipment of ophthalmic surgery, when the practice of «one-day surgery» is becoming increasingly common. It could not only affecting the course of the operation, but also a factor limited the availability of this type of medical care to patients.
Purpose.
To analyze the relationship between the level of anxiety during preoperative preparation and the choice of the optimal approach for the relief of preoperative anxiety in an ophthalmological patient in a «one day» clinic.
Objectives
89 adult patients of both sexes aged from 20 y.o. referred for outpatient surgery (refractive and cataract surgery) and were included into trials, all of them are divided into main - 45 and control - 44 groups. Clinical psychotherapeutic interviewing, a scale of situational and personal anxiety C.D. Spielberger (adapted by Y.L. Khanin), a questionnaire for analyzing satisfaction with the quality of medical services provided in outpatient settings were used. Statistical: Microsoft Excel spreadsheet editor for Windows; the STATISTICA application software package version 6.1. were taken. The condition for determining statistically significant differences is the value р≤0,05.
Methods
Two-stage study by the method of contiuous sampling. Clinical psychotherapeutic interviewing, a scale of situational and personal anxiety C.D. Spielberger (adapted by Y.L. Khanin), a questionnaire for analyzing satisfaction with the quality of medical services provided in outpatient settings were used. Statistical: Microsoft Excel spreadsheet editor for Windows; the STATISTICA application software package version 6.1. were taken. The condition for determining statistically significant differences is the value р≤0,05.
Results
The level of anxiety in patients who received anxiolytics
Level of anxiety
examination day, %
day of operation %
SA
PA
SA
PA
low
22*
22,5
35
*
21
medium
53,5
64,5
57
67
high
24,5*
13
8
*
13
*
- р≤0,05; SA-situational anxiety; PS – personal anxiety
The study revealed an average and high level of situational anxiety in 66% of patients referred for refractive surgery, in 81% of patients referred for cataract surgery. Into main group, against the background of the performed anxiolytic therapy in the preoperative period, the proportion of people with a high level of reactive anxiety decreased significantly (p≤0.05) (from 24% to 8%), while personal anxiety did not significantly change. Among control group patients show a lower level of satisfaction with the quality of medical services provided in an outpatient setting.
Conclusions
The study showed the possibility of providing better quality medical services in «one-day» eye surgery, which expands the availability of outpatient ophthalmic surgical care to patients with high level of anxiety and anxiety disorders.
Disclosure of Interest
None Declared
In photodynamic therapy, light is absorbed by a therapy agent (photosensitizer) to generate reactive oxygen, which then locally kills diseased cells. Here, we report a new form of photodynamic ...therapy in which nonlinear optical interactions of near-infrared laser radiation with a biological medium in situ produce light that falls within the absorption band of the photosensitizer. The use of near-infrared radiation, followed by upconversion to visible or ultraviolet light, provides deep tissue penetration, thus overcoming a major hurdle in treatment. By modelling and experiment, we demonstrate activation of a known photosensitizer, chlorin e6, by in situ nonlinear optical upconversion of near-infrared laser radiation using second-harmonic generation in collagen and four-wave mixing, including coherent anti-Stokes Raman scattering, produced by cellular biomolecules. The introduction of coherent anti-Stokes Raman scattering/four-wave mixing to photodynamic therapy in vitro increases the efficiency by a factor of two compared to two-photon photodynamic therapy alone, while second-harmonic generation provides a fivefold increase.
Nuclear organelles are viscous droplets, created by concentration-dependent condensation and liquid-liquid phase separation of soluble proteins. Nuclear organelles have been actively investigated for ...their role in cellular regulation and disease. However, these studies are highly challenging to perform in live cells, and therefore, their physico-chemical properties are still poorly understood. In this study, we describe a fluorescence lifetime imaging approach for real-time monitoring of protein condensation in nuclear organelles of live cultured cells. This approach unravels surprisingly large cyclic changes in concentration of proteins in major nuclear organelles including nucleoli, nuclear speckles, Cajal bodies, as well as in the clusters of heterochromatin. Remarkably, protein concentration changes are synchronous for different organelles of the same cells. We propose a molecular mechanism responsible for synchronous accumulations of proteins in the nuclear organelles. This mechanism can serve for general regulation of cellular metabolism and contribute to coordination of gene expression.
In this study, we showed that biocompatible zinc oxide (ZnO) nanocrystals (NCs) having a noncentrosymmetric structure can be used as nonresonant nonlinear optical probes for bioimaging applications ...in vitro by use of the second order processes of second harmonic and sum frequency generation, as well as the third order process of four wave mixing. These nonresonant processes provide advantages above and beyond traditional two photon bioimaging: (i) the probes do not photobleach; (ii) the input wavelength can be judiciously selected; and (iii) no heat is dissipated into the cells, ensuring longer cell viability and ultimately longer imaging times. ZnO NCs were synthesized in organic media by using a nonhydrolytic sol−gel process and subsequently dispersed in aqueous media using phospholipid micelles and incorporated with the biotargeting molecule folic acid (FA). Sum frequency, second harmonic, and nonresonant four wave mixing nonlinear signals from this stable dispersion of ZnO NCs, targeted to the live tumor (KB) cells, were used for imaging. Robust intracellular accumulation of the targeted (FA incorporated) ZnO nanocrystals could be observed without any indication of cytotoxicity.
Fixation of biological sample is an essential technique applied in order to “freeze” in time the intracellular molecular content. However, fixation induces changes of the cellular molecular ...structure, which mask physiological distribution of biomolecules and bias interpretation of results. Accurate, sensitive, and comprehensive characterization of changes in biomolecular composition, occurring during fixation, is crucial for proper analysis of experimental data. Here we apply biomolecular component analysis for Raman spectra measured in the same nucleoli of HeLa cells before and after fixation by either formaldehyde solution or by chilled ethanol. It is found that fixation in formaldehyde does not strongly affect the Raman spectra of nucleolar biomolecular components, but may significantly decrease the nucleolar RNA concentration. At the same time, ethanol fixation leads to a proportional increase (up to 40%) in concentrations of nucleolar proteins and RNA, most likely due to cell shrinkage occurring in the presence of coagulant fixative. Ethanol fixation also triggers changes in composition of nucleolar proteome, as indicated by an overall reduction of the α-helical structure of proteins and increase in the concentration of proteins containing the β-sheet conformation. We conclude that cross-linking fixation is a more appropriate protocol for mapping of proteins in situ. At the same time, ethanol fixation is preferential for studies of RNA-containing macromolecules. We supplemented our quantitative Raman spectroscopic measurements with mapping of the protein and lipid macromolecular groups in live and fixed cells using coherent anti-Stokes Raman scattering nonlinear optical imaging.
Recent developments in Raman spectroscopy instrumentation and data processing algorithms have led to the emergence of Ramanomics - an independent discipline with unprecedented capabilities to map the ...distribution of distinct molecular groups in live cells. Here, we introduce a method for probing the absolute concentrations of proteins, RNA and lipids in single organelles of live cultured cells by biomolecular component analysis using microRaman data. We found significant cell-to-cell variations in the molecular profiles of organelles, thus providing a physiologically relevant set of markers of cellular heterogeneity. At the same cell the molecular profiles of different organelles can strongly correlate, reflecting tight coordination of their functions. This correlation was significant in WI-38 diploid fibroblasts and weak in HeLa cells, indicating profound differences in the regulation of biochemical processes in these cell lines.
Using laser tweezers, we study colloidal interactions of solid microspheres in the nematic bulk caused by elastic distortions around the particles with tangential surface anchoring. The interactions ...overcome the Brownian motion when the interparticle separation r-->p is less than 3 particle diameters. The particles attract when the angle theta between r-->p and the uniform far-field director n0 is between 0 degrees and approximately 70 degrees and repel when 75 degrees <or approximately theta<or=90 degrees. The particles aggregate in chains directed at approximately 30 degrees to n0 and, at higher concentrations, form complex kinetically trapped structures.
The paper presents a flux crystal growth technique, studies of the structural peculiarities and the optical absorption spectra of double orthoborate TbCr
3
(BO
3
)
4
with a huntite structure. The ...intensities of the phonon modes were used to determine the ratios of the rhombohedral and monoclinic polytypes for this compound, depending on the growth conditions. The broadband absorption spectra of the Tb
3+
ions in TbCr
3
(BO
3
)
4
single crystals were studied in the temperature range from room temperature to 3.0 K. From them, the energies of the crystal-field levels of the Tb
3+
ion were determined. The temperature dependence of the absorption spectra of the Er
3+
probe ion in TbCr
3
(BO
3
)
4
:Er(1%) shows that there are two phase transitions and agrees with their previously proposed interpretation: at 8.8 K, the chromium subsystem antiferromagnetically orders, and at 5 K, a reorientation of chromium magnetic moments occurs.
To advance an understanding of cellular regulation and function it is crucial to identify molecular contents in cellular organelles, which accommodate specific biochemical processes. Toward ...achievement of this goal, we applied micro-Raman-Biomolecular Component Analysis assay for molecular profiling of major organelles in live cells. We used this assay for comparative analysis of proteins 3D conformation and quantification of proteins, RNA, and lipids concentrations in nucleoli, endoplasmic reticulum, and mitochondria of WI 38 diploid lung fibroblasts and HeLa cancer cells. Obtained data show substantial differences in the concentrations and conformations of proteins in the studied organelles. Moreover, differences in the intraorganellar concentrations of RNA and lipids between these cell lines were found. We report the biological significance of obtained macromolecular profiles and advocate for micro-Raman BCA assay as a valuable proteomics tool.
Fundamental understanding of cellular processes at molecular level is of considerable importance in cell biology as well as in biomedical disciplines for early diagnosis of infection and cancer ...diseases, and for developing new molecular medicine-based therapies. Modern biophotonics offers exclusive capabilities to obtain information on molecular composition, organization, and dynamics in a cell by utilizing a combination of optical spectroscopy and optical imaging. We introduce here a combination of Raman microspectrometry, together with coherent anti-Stokes Raman scattering (CARS) and two-photon excited fluorescence (TPEF) nonlinear optical microscopy, to study macromolecular organization of the nucleus throughout the cell cycle. Site-specific concentrations of proteins, DNA, RNA, and lipids were determined in nucleoli, nucleoplasmic transcription sites, nuclear speckles, constitutive heterochromatin domains, mitotic chromosomes, and extrachromosomal regions of mitotic cells by quantitative confocal Raman microspectrometry. A surprising finding, obtained in our study, is that the local concentration of proteins does not increase during DNA compaction. We also demonstrate that postmitotic DNA decondensation is a gradual process, continuing for several hours. The quantitative Raman spectroscopic analysis was corroborated with CARS/TPEF multimodal imaging to visualize the distribution of protein, DNA, RNA, and lipid macromolecules throughout the cell cycle.
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