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Background Thermoregulation is highly individual and predictive for potentially cascading pathologies. Altered and deficient thermoregulation is considered an important diagnostic indicator which can be of great clinical utility for specialized screening programs and individualized prediction and prevention of severe pathologies triggered early in life. Working hypothesis Individual thermoregulation can be objectively assessed by thermovision camera before and after exercises in school children stratified by age and gender that may be of great clinical utility for personalized training early in life in the framework of 3P medicine. Study design In this study, 60 female and male primary school children were exposed to physical exercises in the form of 45-min general fitness training. The subjects under examination were stratified by age: group 1 (7-year-olds), group 2 (9-year-olds), and group 3 (12-year-olds). Superficial body temperature patterns were measured by means of thermovision camera before and immediately after exercises, as well as after the 15-min recovery time. Temperature patterns were analyzed in 12 areas of the body front and back, covering trunk and upper and lower limbs. Results The obtained results revealed an individual and age-depended difference in response of the body to exercises. The first measurement prior to exercise (measurement 1) revealed no statistically significant differences in the mean surface temperature of all analyzed areas between 7- and 9-year-old children. Further, 7- and 9-year-old children did not differ significantly in the mean temperature recorded in the trunk compared to the 12-year-old children. However, in 12-year-old children, statistically significant higher values of the mean temperature of the upper and lower limbs, were observed compared to the group of 7-year-olds and significantly higher values of the mean temperature of the lower limbs compared to the group of 9-year-olds. Immediately after exercises (measurement 2), a statistically significant decrease in the temperature was noted in all groups and in all areas of the body. The greatest temperature change was observed in 12-year-olds, while the least one was measured in the youngest subjects. The statistically significant relation between the average trunk temperature of 7-year-old and 12-year-old children was observed: lower values of the mean temperature of the front and back of the trunk were noted in the group of 12-year-old children compared to the group of 7-year-olds. A significantly lower average temperature of the back of the trunk compared to the youngest group was also recorded in 9-year-old children. The study performed after the 15-min recovery time (measurement 3) showed an increase in the average temperature of all analyzed areas. In all subjects, the mean temperature recorded in measurement 3 did not differ significantly from the initial ones (measurement 1, prior to exercises). Only the mean temperature of the trunk back of 12-year-old children was significantly lower after the rest period compared to the initial examination. In all groups, the temperatures after exercises followed by a 15-min recovery returned to the initial ones, except of the trunk backs of 12-year-old children, where the temperature was lower than before exercises. Conclusions and expert recommendations in the framework of 3PM Thermovision analysis is an effective tool to assess individual thermoregulation and to stratify school children for personalized exercise coaching. Body exercise-based disease prevention early in life is effective when tailored to the person: multi-parametric guidance for prescribing exercises individually is needed. Contextually, proposed individualized training approach should be adapted to the age-dependent particularities and individual thermoregulation.

Rapid endothelialization of cardiovascular stents is critical to prevent major clinical complications such as restenosis. Reconstruction of the native endothelium on the stent surface can be achieved by the capture of endothelial progenitor cells (EPCs) or neighboring endothelial cells (ECs) in vivo. In this study, stainless steel cardiovascular stents were functionalized with recombinant scFv antibody fragments specific for vascular endothelial growth factor receptor-2 (VEGFR2) that is expressed on EPCs and ECs. Anti-VEGFR2 scFvs were expressed in glycosylated form in Escherichia coli and covalently attached to amine-functionalized, titania-coated steel disks and stents. ScFv-coated surfaces exhibited no detectable cytotoxicity to human ECs or erythrocytes in vitro and bound 15 times more VEGFR2-positive human umbilical vein ECs than controls after as little as 3 min. Porcine coronary arteries were successfully stented with scFv-coated stents with no adverse clinical events after 30 days. Endovascular imaging and histology revealed coverage of the anti-VEGFR2 scFv-coated stent with a cell layer after 5 days and the presence of a neointima layer with a minimum thickness of 80 μm after 30 days. Biofunctionalization of cardiovascular stents with endothelial cell-capturing antibody fragments in this manner offers promise in accelerating stent endothelialization in vivo. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 108B:213-224, 2020.

An increasing interest in a healthy lifestyle raises questions about optimal body weight. Evidently, it should be clearly discriminated between the standardised “normal” body weight and individually optimal weight. To this end, the basic principle of personalised medicine “one size does not fit all” has to be applied. Contextually, “normal” but e.g. borderline body mass index might be optimal for one person but apparently suboptimal for another one strongly depending on the individual genetic predisposition, geographic origin, cultural and nutritional habits and relevant lifestyle parameters—all included into comprehensive individual patient profile. Even if only slightly deviant, both overweight and underweight are acknowledged risk factors for a shifted metabolism which, if being not optimised, may strongly contribute to the development and progression of severe pathologies. Development of innovative screening programmes is essential to promote population health by application of health risks assessment, individualised patient profiling and multi-parametric analysis, further used for cost-effective targeted prevention and treatments tailored to the person. The following healthcare areas are considered to be potentially strongly benefiting from the above proposed measures: suboptimal health conditions, sports medicine, stress overload and associated complications, planned pregnancies, periodontal health and dentistry, sleep medicine, eye health and disorders, inflammatory disorders, healing and pain management, metabolic disorders, cardiovascular disease, cancers, psychiatric and neurologic disorders, stroke of known and unknown aetiology, improved individual and population outcomes under pandemic conditions such as COVID-19. In a long-term way, a significantly improved healthcare economy is one of benefits of the proposed paradigm shift from reactive to Predictive, Preventive and Personalised Medicine (PPPM/3PM). A tight collaboration between all stakeholders including scientific community, healthcare givers, patient organisations, policy-makers and educators is essential for the smooth implementation of 3PM concepts in daily practice.

Quantifying changes in bacteria cells in the presence of antibacterial treatment is one of the main challenges facing contemporary medicine, it is a challenge that is relevant for tackling issues pertaining to bacterial biofilm formation that substantially decreases susceptibility to biocidal agents. Three-dimensional label-free imaging and quantitative analysis of bacteria–photosensitizer interactions, crucial for antimicrobial photodynamic therapy, is still limited due to the use of conventional imaging techniques. We present a new method for investigating the alterations in living cells and quantitatively analyzing the process of bacteria photodynamic inactivation. Digital holographic tomography (DHT) was used for in situ examination of the response of Escherichia coli and Staphylococcus aureus to the accumulation of the photosensitizers immobilized in the copolymer revealed by the changes in the 3D refractive index distributions of single cells. Obtained results were confirmed by confocal microscopy and statistical analysis. We demonstrated that DHT enables real-time characterization of the subcellular structures, the biophysical processes, and the induced local changes of the intracellular density in a label-free manner and at sub-micrometer spatial resolution.

Current vascular stents, such as drug eluting stents (DES), have some serious drawbacks, like in stent restenosis and thrombosis. Therefore, other solutions are sought to overcome these post-implantations complications. These include the strategy of biofunctionalization of the stent surface with antibodies that facilitate adhesion of endothelial cells (ECs) or endothelial progenitor cells (EPCs). Rapid re-endothelialization of the surface minimizes the risk of possible complications. In this study, we proposed ammonium acryloyldimethyltaurate/vinylpyrrolidone co-polymer-based surface (AVC), which was mercaptosilanized in order to expose free thiol groups. The presence of free thiol groups allowed for the covalent attachment of CD133 antibodies by disulfide bridges formation between mercaptosilanized surface and cysteine of the protein molecule thiol groups. Various examinations were performed in order to validate the procedure, including attenuated total reflection&ndash, Fourier transform infrared spectroscopy (ATR-FTIR) and Fourier transform Raman spectroscopy (FT-Raman), atomic force microscopy (AFM) and scanning electron microscopy (SEM). By means of ATR-FTIR spectroscopy presence of the CD133 antibody within coating was confirmed. In vitro studies proved good biocompatibility for blood cells without induction of hemolytic response. Thus, proposed biofunctionalized CD133 antibody AVC surface has shown sufficient stability for adapting as cardiovascular implant coating and biocompatibility. According to conducted in vitro studies, the modified surface can be further tested for applications in various biological systems.

Supporting role of silica nanospheres with surface modified by silver nanoparticles in antibacterial photodynamic therapy (APDT), was studied. Optical and antibacterial properties of a photosensitizer – Photolon® in the presence of silica based nanopowders doped with silver nanoparticles, were determined. Various concentrations of Photolon® solution and nanomaterials have been prepared in order to examine the optical absorption and the effectivity of antibacterial activity. It was found that the maximum optical absorption of Photolon® depends on its concentration and can be enhanced by the presence of silver nanoparticles. The presence of silver nanoparticles can thus lead to a reduction of the concentration of the photosensitizer while achieving the same level of antibacterial efficacy as in case of higher photosensitizer concentration without the presence of silver nanoparticles. Efficiency of APDT was enhanced by combination of nanosilver doped silica nanospheres, which have powerful antimicrobial and bactericidal activity and photoactive photosensitizer – Photolon®.

Cellulite, the problem of dimpled appearance of the skin, affects approximately 85% of female population in developed countries and is classified as one of the worst tolerated by women deteriorating their quality of life and self-esteem. There is a lack of early, objective, quantitative and personalized diagnosis of different stages of cellulite, thus making prevention or early therapeutic intervention difficult. We have demonstrated the efficacy of thermal imaging using IR thermography in a group of female volunteers with different stages of cellulite. By analyzing the superficial temperature distribution of the body, it was possible to diagnose the cellulite stage. The thermal images of posterior site of thighs were recorded, and cellulite areas were identified for further quantitative analysis. We used a custom-designed classification scheme for automatic recognition of the different stages of cellulite as per the well-known Nurnberger–Muller diagnosis scheme. It was possible to diagnose the cellulite stages with over 80% accuracy. The accuracy can be further increased to over 97% using a threshold value correction scheme. Our work has shown that IR thermography when coupled with computer-aided imaging analysis and processing can be a very convenient and effective tool to enable personalized diagnosis and preventive medicine to improve the quality of life of women with cellulite problem.

In this study, the infrared thermography was used to examine heating effects in human auricle region due to short-duration use of smartphones in normal and flight mode operation by 20 healthy volunteers (age 19–24 years, 9 females and 11 males). Statistically significant differences in both modes were observed between mean temperatures of the auricle regions before and immediately after the use of the smartphone. The increase in tissue temperature resulted from a combined effect of conductive heat transfer caused by the non-microwave-related activity of the smartphone and the absorption of the radiofrequency microwaves. Furthermore, we observed the thermal effect even after 1 h of such short use of the smartphone. The transient temperature change in tissues due to smartphone use can contribute to the cumulative thermal effects and thus both short- and long-duration use of smartphones should be considered in future studies. The small study also showed a gender dimension in the thermal response, which also merits further investigation. The study confirmed the usefulness of using thermography for direct quantitative analyses of temperature changes related to smartphone usage, which can be exploited in future epidemiological studies in personalized and preventive medicine.

Thermal imaging is a safe, noninvasive, and quite low-cost technique that allows for the rapid and noninvasive recording of thermal radiation emitted by the human body. In recent years, it is increasingly used for physiotherapy monitoring. The main goal of the study was to compare the influence of two various treatments applied in two various orders, namely local cryotherapy (CRYO) and ultrasound therapy (US). The impact of combined therapies, applied in two various orders, was studied in terms of changes of the superficial temperature distribution on the front right thigh surface. The response to the therapy depending the treated persons’ gender, along with a consensual reaction, was examined. The research material consisted of 30 healthy volunteers, 19–23-year old students, aged 19.93 (± 1.42). All treatments were performed at weekly intervals in the following order: in the first week, CRYO immediately followed by US, and in the second week, ultrasound immediately followed by local cryotherapy. All treatments were performed on the right quadriceps. The combined sequence of local cryotherapy and ultrasound therapy caused temperature decrease in the treated area regardless of their performance order. The temperature decrease considerably depends on the sequence of the applied treatments, and it was more significant after the ultrasound–cryotherapy sequence. Moreover, these changes also lasted longer. Statistically significant influence of the subjects’ gender was observed in case of consensual reaction.

Background Contemporary medicine does not concern the issue of dosimetry in photodynamic diagnosis (PDD) but follows the photosensitizer (PS) producers recommendation. Most preclinical and clinical PDD studies indicate a considerable variation in the possibility of visualization and treatment, as e.g. in case of cervix lesions. Although some of these variations can be caused by the different histological subtypes or various tumor geometries, the issue of varying PS concentration in the tumor tissue volume is definitely an important factor. Therefore, there is a need to establish the objective and systematic PDD dosimetry protocol regarding doses of light and photosensitizers. Methods Four different irradiation sources investigated in PDD (literature) were used for PS excitation. The PS luminescence was examined by means of the non-imaging (spectroscopic) and imaging (wide- and narrow-field of view) techniques. The methodology for low-level intensity photoluminescence (PL) characterization and dedicated image processing algorithm for PS luminescence images analysis were proposed. Further, HeLa cells' cultures penetration by PS was studied by a confocal microscopy. Results Reducing the PS dose with the choice of proper photoexcitation conditions decreases the PDD procedure costs and the side effects, not affecting the diagnostic efficiency. We determined in vitro the minimum incubation time and photosensitizer concentration of Photolon for diagnostic purposes, for which the Photolon PL can still be observed. It was demonstrated that quantification of PS concentration, choice of proper photoexcitation source, appropriate adjustment of light dose and PS penetration of cancer cells may improve the low-level luminescence photodynamic diagnostics performance. Conclusions Practical effectiveness of the PDD strongly depends on irradiation source parameters (bandwidth, maximum intensity, half-width) and their optimization is the main conditioning factor for low-level intensity and low-cost PDD.

In this study we present the porous silica-based material that can be used for in situ drug delivery, offering effective supply of active compounds regardless its water solubility. To demonstrate usability of this new material, three silica-based materials with different pore size distribution as a matrix for doping with Photolon (Ph) and Protoporphyrin IX (PPIX) photosensitizers, were prepared. These matrices can be used for coating cardiovascular stents used for treatment of the coronary artery disease and enable intravascular photodynamic therapy (PDT), which can modulate the vascular response to injury caused by stent implantation&mdash, procedure that should be thought as an alternative for drug eluting stent. The FTIR spectroscopic analysis confirmed that all studied matrices have been successfully functionalized with the target photosensitizers. Atomic force microscopy revealed that resulting photoactive matrices were very smooth, which can limit the implantation damage and reduce the risk of restenosis. No viability loss of human peripheral blood lymphocytes and no erythrocyte hemolysis upon prolonged incubations on matrices indicated good biocompatibility of designed materials. The suitability of photoactive surfaces for PDT was tested in two cell lines relevant to stent implantation: vascular endothelial cells (HUVECs) and vascular smooth muscle cells (VSMC). It was demonstrated that 2 h incubation on the silica matrices was sufficient for uptake of the encapsulated photosensitizers. Moreover, the amount of the absorbed photosensitizer was sufficient for induction of a phototoxic reaction as shown by a rise of the reactive oxygen species in photosensitized VSMC. On the other hand, limited reactive oxygen species (ROS) induction in HUVECs in our experimental set up suggests that the proposed method of PDT may be less harmful for the endothelial cells and may decrease a risk of the restenosis. Presented data clearly demonstrate that porous silica-based matrices are capable of in situ delivery of photosensitizer for PDT of VSMC.

Background Cellulite is a common physiological condition of dermis, epidermis, and subcutaneous tissues experienced by 85 to 98% of the post-pubertal females in developed countries. Infrared (IR) thermography combined with artificial intelligence (AI)-based automated image processing can detect both early and advanced cellulite stages and open up the possibility of reliable diagnosis. Although the cellulite lesions may have various levels of severity, the quality of life of every woman, both in the physical and emotional sphere, is always an individual concern and therefore requires patient-oriented approach. Objectives The purpose of this work was to elaborate an objective, fast, and cost-effective method for automatic identification of different stages of cellulite based on IR imaging that may be used for prescreening and personalization of the therapy. Materials and methods In this study, we use custom-developed image preprocessing algorithms to automatically select cellulite regions and combine a total of 9 feature extraction methods with 9 different classification algorithms to determine the efficacy of cellulite stage recognition based on thermographic images taken from 212 female volunteers aged between 19 and 22. Results A combination of histogram of oriented gradients (HOG) and artificial neural network (ANN) enables determination of all stages of cellulite with an average accuracy higher than 80%. For primary stages of cellulite, the average accuracy achieved was more than 90%. Conclusions The implementation of computer-aided, automatic identification of cellulite severity using infrared imaging is feasible for reliable diagnosis. Such a combination can be used for early diagnosis, as well as monitoring of cellulite progress or therapeutic outcomes in an objective way. IR thermography coupled to AI sets the vision towards their use as an effective tool for complex assessment of cellulite pathogenesis and stratification, which are critical in the implementation of IR thermographic imaging in predictive, preventive, and personalized medicine (PPPM).

Background: Liposomes serve as delivery systems for biologically active compounds. Existing technologies inefficiently encapsulate large hydrophilic macromolecules, such as PVP-conjugated chlorin e6 (Photolon). This photoactive drug has been widely tested for therapeutic applications, including photodynamic reduction of atherosclerotic plaque. Methods: A novel formulation of Photolon was produced using &ldquo, gel hydration technology&rdquo, Its pharmacokinetics was tested in Sus scrofa f. domestica. Its cellular uptake, cytotoxicity, and ability to induce a phototoxic reaction were demonstrated in J774A.1, RAW264.7 macrophages, and vascular smooth muscle (T/G HA-VSMC) as well as in vascular endothelial (HUVEC) cells. Results: Developed liposomes had an average diameter of 124.7 &plusmn, 0.6 nm (polydispersity index (PDI) = 0.055) and contained &gt, 80% of Photolon). The half-life of formulation in S. scrofa was 20 min with area under the curve (AUC) equal to 14.7. The formulation was noncytotoxic in vitro and was rapidly (10 min) and efficiently accumulated by macrophages, but not T/G HA-VSMC or HUVEC. The accumulated quantity of photosensitizer was sufficient for induction of phototoxicity in J774A.1, but not in T/G HA-VSMC. Conclusions: Due to the excellent physical and pharmacokinetic properties and selectivity for macrophages, the novel liposomal formulation of Photolon is a promising therapeutic candidate for use in arteriosclerosis treatment when targeting macrophages but not accompanying vascular tissue is critical for effective and safe therapy.

The proper lipid composition of liposomes designed to carry drugs determines their surface properties ensuring their accumulation within selected tissue. The electrostatic potential and surface topology of liposomes affect the internalization by single cells. The high-resolution imaging of cancer cells and the distribution of protoporphyrin-loaded liposomes within the cytoplasm and its dependence on the liposome surface properties are presented. In the paper, HeLa cells were used to investigate the uptake of porphyrin-loaded liposomes and liposomes alone by means of confocal and differential interference contrast microscopies. The effect of liposomes surface electrostatic potential and surface topology on their intracellular distribution was evaluated. The time evolution of the intracellular distribution of liposomes labelled with Rhodamine-PE was examined on HeLa cells. These studies allow for the identification of the liposome lipid composition so the efficient delivery of the active substance to cancer cells will be achieved. The obtained results showed that neutral PC-liposomes are the most efficiently internalized by HeLa cells. Moreover, results showed that properties of liposomes affect not only the internalization efficiency of the photosensitizer but also its distribution within the cells, as revealed by colocalization measurements.

The development of new diagnostics techniques and modalities is critical for early detection of microbial contamination. In this study, the novel integrated system for multi-parametric optical phenotyping and characterization of bacterial colonies, is presented. The system combines Mach-Zehnder interferometer with a spectral imaging system for capturing multispectral diffraction patterns and multispectral two-dimensional transmission maps of bacterial colonies, along with the simultaneous interferometric profilometry. The herein presented investigation was carried out on five representative bacteria species and nearly 3000 registered multispectral optical signatures. The interferograms were analyzed by four-step phase shift algorithm to reconstruct the colony profile to enable the obtaining of the comparable optical signatures. The dedicated image processing algorithms were used for extraction of quantitative features of these signatures. The random forest algorithm was applied for selection of the most predictive set of features, which were used in classification model based on Support-Vector Machine. Obtained results have shown that the use of multiple multispectral optical signatures provide a multi-parametric bacteria identification at an exceptionally high accuracy (99.4–100%), significantly better than in case of classification based on each of these signatures (multispectral diffraction patterns, two-dimensional transmission coefficient maps), separately. Obtained results revealed that analysis of multispectral signatures can also be applied for characterisation of physical, physicochemical and chemical properties of the bacterial colonies in the presence of the antimicrobial factors. Therefore, the proposed label-free, non-destructive optical technique has perspectives to be exploited in the multipurpose diagnostics and it can be used as a pre-screening tool in microbiological laboratories.

In our study, we describe the outcomes of the intercalation of different anthracycline antibiotics in double-stranded DNA at the nanoscale and single molecule level. Atomic force microscopy analysis revealed that intercalation results in significant elongation and thinning of dsDNA molecules. Additionally, using optical tweezers, we have shown that intercalation decreases the stiffness of DNA molecules, that results in greater susceptibility of dsDNA to break. Using DNA molecules with different GC/AT ratios, we checked whether anthracycline antibiotics show preference for GC-rich or AT-rich DNA fragments. We found that elongation, decrease in height and decrease in stiffness of dsDNA molecules was highest in GC-rich dsDNA, suggesting the preference of anthracycline antibiotics for GC pairs and GC-rich regions of DNA. This is important because such regions of genomes are enriched in DNA regulatory elements. By using three different anthracycline antibiotics, namely doxorubicin (DOX), epirubicin (EPI) and daunorubicin (DAU), we could compare their detrimental effects on DNA. Despite their analogical structure, anthracyclines differ in their effects on DNA molecules and GC-rich region preference. DOX had the strongest overall effect on the DNA topology, causing the largest elongation and decrease in height. On the other hand, EPI has the lowest preference for GC-rich dsDNA. Moreover, we demonstrated that the nanoscale perturbations in dsDNA topology are reflected by changes in the microscale properties of the cell, as even short exposition to doxorubicin resulted in an increase in nuclei stiffness, which can be due to aberration of the chromatin organization, upon intercalation of doxorubicin molecules.

We report a multistep strategy of biochemical surface modifications that resulted in the synthesis of new, effective and biocompatible intravascular implants coating with immobilized anti-CD133 antibodies, that proved to be the most effective in endothelial progenitor cells capture and reduced smooth muscle cells growth. Biomolecules were immobilized on differently functionalized surfaces. The distribution, nanostructural characteristics and intramolecular interactions of anti-CD133 molecules as well as their ability to bind EPCs was evaluated. We also tempted to build a molecular model of the CD133 protein to study antigen-antibody interactions. CD133 protein is expressed in endothelial progenitor cells (EPCs). Absence of preferential interaction site on CD133, but rather a presence of a small binding area, may be the specificity of reconnaissance sequence, thus importantly increasing the probability of CD133 protein binding. After all, regarding our molecular model, we are convinced that specific, and large enough interactions between anti-CD133 coating stent surface and CD133 present on EPCs will reduce risk of restenosis by favoring the endothelial growth. Additionally, the safety study of the vivo performance of modified titania based surface was performed using small animal models. No allergological or toxical local or systemic adverse effects of the developed coatings were noted.

Stainless steel 316 L material is commonly used for the production of coronary and peripheral vessel stents. Effective biofunctionalization is a key to improving the performance and safety of the stents after implantation. This paper reports the method for the immobilization of recombinant antibody fragments (scFv) on stainless steel 316 L to facilitate human endothelial progenitor cell (EPC) growth and thus improve cell viability of the implanted stents for cardiovascular applications. The modification of stent surface was conducted in three steps. First the stent surface was coated with titania based coating to increase the density of hydroxyl groups for successful silanization. Then silanization with 3 aminopropyltriethoxysilane (APTS) was performed to provide the surface with amine groups which presence was verified using FTIR, XPS, and fluorescence microscopy. The maximum density of amine groups (4.8*10(-5) mol/cm(2)) on the surface was reached after reaction taking place in ethanol for 1 h at 60 °C and 0.04M APTS. On such prepared surface the glycosylated scFv were subsequently successfully immobilized. The influence of oxidation of scFv glycan moieties and the temperature on scFv coating were investigated. The fluorescence and confocal microscopy study indicated that the densest and most uniformly coated surface with scFv was obtained at 37 °C after oxidation of glycan chain. The results demonstrate that the scFv cannot be efficiently immobilized without prior aminosilanization of the surface. The effect of the chemical modification on the cell viability of EPC line 55.1 (HucPEC-55.1) was performed indicating that the modifications to the 316 L stainless steel are non-toxic to EPCs.

The condensing effect of aminoglycoside antibiotics on the structure of double-stranded DNA was examined. The selective condensation of DNA by small molecules is an interesting approach in biotechnology. Here, we present the interaction between calf thymus DNA and three types of antibiotic molecules: tobramycin, kanamycin, and neomycin. Several techniques were applied to study this effect. Atomic force microscopy, transmission electron microscopy images, and nuclear magnetic resonance spectra showed that the interaction of tobramycin with double-stranded DNA caused the rod, toroid, and sphere formation and very strong condensation of DNA strands, which was not observed in the case of other aminoglycosides used in the experiment. Studies on the mechanisms by which small molecules interact with DNA are important in understanding their functioning in cells, in designing new and efficient drugs, or in minimizing their adverse side effects. Specific interactions between tobramycin and DNA double helix was modeled using molecular dynamics simulations. Simulation study shows the aminoglycoside specificity to bend DNA double helix, shedding light on the origins of toroid formation. This phenomenon may lighten the ototoxicity or nephrotoxicity issues, but also other adverse reactions of aminoglycoside antibiotics in the human body. Electronic supplementary material The online version of this article (doi:10.1007/s00249-015-1095-9) contains supplementary material, which is available to authorized users.

Most physical treatments have thermal effects. Effect of heat on the body is not always uniform and depends on the treated body surface, the intensity of the stimulus, the stimulus duration, changes in the intensity of the stimulus and the physical environment. Thermovision is a method that allows for precise determination of the temperature distribution of the tested body surface. The aim of the study was to evaluate the effect of sequence of execution of infrared irradiation combined with visible light, for simplicity called further IR and diadynamic currents (called further DD) on the distribution of skin surface temperature. We also evaluated whether the order of execution of physical treatments affects the consensual reaction and temperature variations depending on sex and body mass index. The study involved 28 volunteers—16 women and 12 men aged 19–28 years. The treatments were performed on the front of right thigh in two sequences—IR–DD and DD–IR. For measuring the temperature distribution, there was thermovision used. In each sequence, there were five images captured with the use of FLIR T335 camera. The significance of changes in skin temperature was rated with Student’s t test for dependent samples, whereas the dependence on the treatments sequence and sexual differentiation was analyzed with the use of test for independent samples, and the correlations were evaluated by determining the Pearson’s r linear correlation coefficient. In the assessment of statistical significance, the critical threshold α = 0.05 was used. Regardless of the treatments sequence, the average surface temperature of the right thigh after IR radiation increased by more than 4 °C. However, there were no significant temperature changes observed after the treatment with diadynamic currents. More dynamic changes in the average temperature values were stated in the sequence DD–IR, as 30 min after the second treatment, the temperature was higher as compared to the initial value by 1 °C. Statistically significant sex-dependent differences occurred only after infrared radiation.

Stainless steel 316L is a material commonly used in cardiovascular medicine. Despite the various methods applied in stent production, the rates of in-stent restenosis and thrombosis remain high. In this study graphene was used to coat the surface of 316L substrate for enhanced bio- and hemocompatibility of the substrate. The presence of graphene layers applied to the substrate was investigated using cutting-edge imaging technology: energy-filtered low-voltage FE-SEM approach, scanning electron microscopy (SEM), Raman spectroscopy, and atomic force microscopy (AFM). The potential of G-316L surface to influence endothelial cells phenotype and endothelial-to-mesenchymal transition (EndoMT) has been determined. Our results show that the bio- and hemocompatible properties of graphene coatings along with known radial force of 316L make G-316L a promising candidate for intracoronary implants.

A lack of biocompatibility in stents could lead to adverse biological effects. This chapter provides an overview of different strategies for physicochemical stent functionalization aimed at reducing the occurrence of thrombosis and restenosis and accelerating rapid endothelialization after stent implantation. In some strategies, a layer of material is introduced on the stent surface in order to prohibit or reduce undesirable protein adhesion. The substances used in this manner are polymers, metal oxides, and gold. Other approaches involve direct attachment of endothelial progenitor cells and antibody fragments to the stent surface. For metallic samples, plasma polymerization technique has been applied for effective covalent protein attachment. Lastly, there is the strategy of stent surface functionalization with thiol groups. Despite the number of studies on stent surface functionalization for its improved biocompatibility, the ultimate design remains a challenge. There is a great potential in future research for developing a stent material with increased detection sensitivities for endothelial cells and hence acceleration of the healing process after stent implantation.

Mechanical properties of biological structures play an important role in regulating cellular activities and are critical for understanding metabolic processes in cancerous cells and the effects of drugs. For some cancers, such as acute myeloid leukaemia, chemotherapy is one of preferential methods. However, due to the lack of selectivity to cancer cells, cytostatic agents cause toxicity to normal tissues. Here, we study the effect of doxorubicin (DOX) on the mechanical properties of DNA molecules, leukemic blast cells and erythrocytes, using optical tweezers. In addition, we controlled the subcellular distribution of the drug by confocal microscopy. Our results indicated that doxorubicin affects mechanical properties of cellular structures. In all cases the drug reduced mechanical strength of examined objects. For the leukemic cells the drug subcellular distribution was predominantly nuclear with some particulate cytoplasmic fluorescence. In erythrocytes, doxorubicin showed fluorescence mainly in cytoplasm and plasma membrane. The lowering of blast cells stiffness may be due to the interaction of doxorubicin with nuclear structures, especially with nucleic acids, as our studies with DNA confirmed. In addition, it is known that DOX inhibits the polymerization of actin and thus cytoskeletal modification may also be important in reducing of cell mechanical strength. In the case of erythrocytes - the non-nucleated cells, a significant effect on the decrease of cell stiffness, besides the cytoskeleton, may have the interaction of the drug with the cell membrane. Experiments with model phospholipid membranes confirmed that observed increase in cell elasticity originates, among other things, from the drug incorporation in the lipid membrane itself. The lowering of mechanical strength of leukemic cells may have an significant impact on the effectiveness of chemotherapy. However, the fact that doxorubicin interacts not only with proliferating cancer cells, but also with the health ones may explains the high toxicity of the drug at the therapeutic concentrations. Our observations also suggest that chemotherapy with doxorubicin may decrease the risk of vascular complications in acute leukemia, due to increasing the cell elasticity.

Intravascular stenting of atherosclerotic coronary arteries is a life-saving, widely used procedure in interventional cardiology. Adverse clinical outcomes such as restenosis highlight the importance of meeting the excellent biocompatibility by cardiovascular implants. Many attempts have been made to improve the safety profile of implant surface. We for the first time developed the photoactive intravascular titania-based nanomaterials for the application as cardiovascular stent coating. Photoactive biomaterial deposited on the cardiovascular stent surface demonstrated promising features, making it an excellent substrate for endothelial cells growth and proliferation. The biocompatibility of these coatings has been compared with 316L stainless steel surfaces typically used in commercial coronary stents production. The results of the study proved that the innovative titania-based coatings have better biocompatibility characteristics than the 316L stainless steel and in regard of its antithrombotic potential provided protection against restenosis. Furthermore, the titania coating supported endothelial cells attachment and proliferation, and induced prolonged plasma recalcification time in comparison with stainless steel surface. Innovative photoactive titania coating can be an important factor to prevent the process of the restenosis in the place of implantation.

Methods based on light diffraction on bacterial colonies create the new perspectives for innovations in microbiological diagnosis. Contrary to the conventional techniques, the proposed optical methods are non-contact and non-destructive. There is no requirement for any additional chemicals, immunological or fluorescence markers. The proposed methods are based on the images of light diffraction on bacterial colonies grown on solid nutrient media analyzed by statistical methods. The application of the digital holography can additionally facilitate and improve the efficiency of the microbiological examination.

The aim was to model the influence of LASIK surgery on the optical parameters of the human eye and to ascertain which factors besides the central corneal radius of curvature and central thickness play the major role in postsurgical refractive change. Ten patients were included in the study. Pre- and postsurgical measurements included standard refraction, anterior corneal curvature and pachymetry. The optical model used in the analysis was based on the Le Grand and El Hage schematic eye, modified by the measured individual parameters of corneal geometry. A substantial difference between eye refractive error measured after LASIK and estimated from the eye model was observed. In three patients, full correction of the refractive error was achieved. However, analysis of the visual quality in terms of spot diagrams and optical transfer functions of the eye optical system revealed some differences in these measurements. This suggests that other factors besides corneal geometry may play a major role in postsurgic...

Photocatalytic and in situ microbial activity of the amorphous and annealed states of Ag-doped and un-doped titania were examined. Studies on their structure, morphology, composition, and the photo-absorption characteristics of these materials were performed. These results were correlated with the photocatalytic and microbial activity against methicillin resistant Staphylococcus aureus K324 (MRSA), methicillin susceptible S. aureus ATCC 25923 (MSSA), Escherichia coli PA 170, and yeasts Candida albicans ATCC 90028. The annealed powders containing anatase form of titania exhibited relatively higher photocatalytic activity, corresponding to activity against MRSA, when exposed to UV-A radiation. In comparison, amorphous powders exhibited low photoactivity and showed poor antibacterial performance against MRSA under UV-A exposure. Doping of amorphous titania with Ag resulted in an anti-MRSA effect without exposure to UV radiation. In the Ag-doped crystalline anatase samples, the size of Ag primary nanocrystallites increased, which led to the decrease in the surface concentration of Ag and detriment anti-MRSA activity.

Nanotechnology for biomedical applications - enhancement of photodynamic activity by nanomaterialsOver the last two decades nanotechnology has become one of the most dynamically evolving field of research. The unique properties of nanomaterials, not disclosing at microscale, are examined and exploited to extend our understanding of the interactions taking place at atomic or molecular level. Those findings affect research in many areas, like e.g. alternative energy sources, electronics, physics and medicine. In this paper, the possibility of using nanomaterials for the enhancement of photodynamic activity, is discussed. A brief review on drug-delivery facilitating nanomaterials, regarding their characteristic features, is presented. An exemplary application of silver-doped nanomaterials for enhancement of photodynamic properties of two photosensitizers: Photolon and Protoporphyrin IX, is described. Influence of silver-doped nanomaterials addition on the fluorescence intensity of photosensitizers immobilized in silica-titania (SiO2-TiO2) sol was examined via VIS spectroscopy. Influence of sonication on the fluorescence enhancement was also investigated. It was demonstrated that the fluorescence enhancement of photosensitizers depends on the concentration of both: photosensitizer and silver-doped nanoparticles.

The potential use of a novel multichannel optical system towards fast and non-destructive bacteria identification and its application for environmental bacteria characterisation on the strain level is presented. It is the first attempt to use the proposed optical method to study various bacteria species (Gram-negative, Gram-positive) commonly present in the environment. The novel configuration of the optical system enables multichannel examination of bacterial colonies and provides additional functionality such as registration of two-dimensional (2D) distribution of monochromatic transmission coefficient of examined colonies, what can be used as a novel optical signature for bacteria characterization. Performed statistical analysis indicates that it is possible to identify representatives of environmental soil bacteria on the species level with the 98.51% accuracy and in case of two strains of Rahnella aquatilis bacteria on the strain level with the 98.8% accuracy. The proposed method is an alternative to the currently used preliminary bacteria examination in environment safety control with the advantage of being fast, reliable, non-destructive and requiring minimal sample preparation.

In view of laser-assisted medical applications, the construction of silica-based sol-gel fiberoptic sensors based on photolon (Ph) and protoporphyrin IX (PP IX) is discussed. Electron microscopy and AFM were used to characterize the silica sol-gel coatings. AFM measurements indicate a change in the surface porosity. The PP IX-based sensors were constructed as a one-layer optode as well as a multilayered structure. An additional hybrid sensor made up of alternate layers of PP IX-and Ph-doped sol-gel was also constructed and examined. Sol-gel matrices were prepared from silicate precursor tetraethylorthosilicate (TEOS) mixed with ethanol in acid-catalyzed hydrolysis. The carrier matrices of photosensitive dyes were produced with factor R = 20, where R denotes the ratio of solvent moles (ethanol) to the number of TEOS moles. A multilayered coating was built up using the reverse-dipping technique. The overall coating thickness was determined by electron microscopy. Doped sol-gels with different PP IX concentrations were used to produce fiberoptic coatings. The film optodes with a different number of layers were examined by fluorescence spectroscopy. It was found that photolon and protoporphyrin IX entrapped in sol-gel preserve their chemical reactivity and have contact with the external environment. The hybrid sensor demonstrated clear fluorescence and a reversible behavior in gaseous environments.

The use of light diffraction for the microbiological diagnosis of bacterial colonies was a significant breakthrough with widespread implications for the food industry and clinical practice. We previously confirmed that optical sensors for bacterial colony light diffraction can be used for bacterial identification. This paper is focused on the novel perspectives of this method based on digital in-line holography (DIH), which is able to reconstruct the amplitude and phase properties of examined objects, as well as the amplitude and phase patterns of the optical field scattered/diffracted by the bacterial colony in any chosen observation plane behind the object from single digital hologram. Analysis of the amplitude and phase patterns inside a colony revealed its unique optical properties, which are associated with the internal structure and geometry of the bacterial colony. Moreover, on a computational level, it is possible to select the desired scattered/diffracted pattern within the entire observation volume that exhibits the largest amount of unique, differentiating bacterial features. These properties distinguish this method from the already proposed sensing techniques based on light diffraction/scattering of bacterial colonies. The reconstructed diffraction patterns have a similar spatial distribution as the recorded Fresnel patterns, previously applied for bacterial identification with over 98% accuracy, but they are characterized by both intensity and phase distributions. Our results using digital holography provide new optical discriminators of bacterial species revealed in one single step in form of new optical signatures of bacterial colonies: digital holograms, reconstructed amplitude and phase patterns, as well as diffraction patterns from all observation space, which exhibit species-dependent features. To the best of our knowledge, this is the first report on bacterial colony analysis via digital holography and our study represents an innovative approach to the subject.

The surface passivation with titanium sol-gel coatings is a frequently used technique to control the adsorption of selected biological macromolecules and to reduce the exposure of the bulk material to biological matter. Due to the increasing number of new coating-preparation methods and new gel compositions with various types of additives, the quality and homogeneity determination of the surface covering is a critical factor affecting performance of any implanted material. While coating thickness is easy to determine, the homogeneity of the surface distribution of coating materials requires more elaborate methodologies. In the paper, the laser induced breakdown spectroscopy (LIBS) based method, capable to quantitate the homogeneity and uniformity of the europium in titanium dioxide sol-gel coatings on stainless steel surfaces prepared with two different procedures: spin-coating and dip-coating, is presented. The emission intensity of titanium has been used to determine the coating thickness whereas the relative values of europium and titanium emission intensities provide data on the coating homogeneity. The obtained results show that the spin-coating technique provides better surface coverage with titanium dioxide. However, when the surface coating compositions were compared the dip-coating technique was more reliable.

The study was carried out on Escherichia coli strains isolated from poultry and cows. The bacteria were incubated and treated by Photolon, a plant-based photosensibilizator, and exposed after that to laser light at 662 nm. Additionally, the photosensitizer was immobilized in fiberoptic sol–gel coating, constructed as an 8-layer applicator. It was demonstrated that Photolon inhibits bacteria growth. Photolon entrapped into the sol–gel matrix also exhibits photodynamic activity and no bacteria were seen near the doped sol–gel applicator.

A new type of applicator for interstitial laser coagulation is proposed in this paper. The new fiber optic applicator is made by removing the original fiber cladding and replacing it with a sol-gel coating. The sol-gel coating was prepared from the silicate precursor TEOS (tetraethylorthosilicate), mixed with ethanol in acid, which catalyzes hydrolysis. The material is produced with the factor R = 20, where R denotes the number of solvent moles to the number of TEOS moles. In these studies, optical fibers were used from Laser Components (core diameter 400 µm, HCS, low OH). The external jacket was mechanically removed, leaving a remaining 2-cm length of bare fiber. The modified dip-coating method was used to cover the bare fibers with sol-gel material. Two types of applicators were produced: pure sol-gel coated devices and applicators with sol-gel coating doped with photosensitive dyes. The photodynamic activity of chlorophyll-based Photolon and Protoporphyrin IX (PPIX) incorporated within the sol-gels was studied spectrophotometrically. It was demonstrated that sol-gel applicators are capable of performing laser interstitial coagulation using a semiconductor cw laser at 980 nm. It was also demonstrated that the incorporated photosensibilizers retain their photochemical activity.

In the past years new materials have been developed for innovative applications in the field of medical technology. The sol–gel-coated applicators were prepared from the silicate precursor tetraethylorthosilicate (TEOS) mixed with the solvent ethanol in acid to catalyze hydrolysis. The sol–gel layers were deposited on the fiber optic core, substituting the original fiber coating which had been removed. In these studies 2 m long optical fibers were used from laser components (core diameter 400 μm, HCS, low OH). Two types of applicators were produced with 2 and 3 cm long tips, respectively. The sol–gel coatings were prepared with factor R = 20 , the R denoting the number of alcohol moles to the number of TEOS moles. All applicators are produced as eight-layer applicators, denoting that eight single layers of sol–gel material were placed successively onto the pre-cleaned bare fiber. Examinations were carried out to investigate the optical properties of the diffusely emitting applicators. A diode laser (λ=980 nm) in continuous as well as in pulsed mode was used for interstitial coagulation in vitro. Excellent results were obtained in view of the simple and low cost preparation procedure for the sol–gel-coated applicators. It was found that the size of coagulated tissue depends on the applicator length, exposure time and laser power.

The suitability of the silica sol–gel based coatings for the construction of sol–gel fiberoptics applicators for interstitial light therapy was analyzed in this study. The various applicators were developed: bare core, tapered and so-called ‘zebra’ type. The sol–gel layers were placed on bare fiber core by dip-coating or painting techniques. Various materials were tested: silica sol–gels produced with molar ratios R =5, 15, 32, 50, where molar ratio denotes the number of solvent moles to the number of precursor moles. In this study tetraethoxysilane as a precursor and ethyl alcohol as a solvent were used. The distribution of light scattered from the applicator was examined. It was proved that the radiation pattern strongly depends on the material characteristic ( R ratio), shape of the fiber tip and number of sol–gel coating layers. The most intensive scattering was observed in the case of sol–gel material produced with R =5.