[Study of the growth temperature of ocular surface microorganisms in norm and in infectious keratitis]. / Izuchenie temperaturnykh uslovii rosta mikroorganizmov glaznoi poverkhnosti v norme i pri infektsionnykh keratitakh.

Standard bacteriological examinations, which involve culturing microorganisms at 37 °C, are commonly used in clinical practice for diagnosing infectious diseases. However, the growth temperature of microorganisms on the ocular surface (OS) during infectious keratitis (IK) may not coincide with the laboratory standard, which is due to the characteristic features of heat exchange in the eye. PURPOSE: This exploratory study examines the distribution and properties of OS microorganisms isolated under different temperature cultivation conditions in patients with IK and healthy volunteers without ophthalmic pathology. MATERIAL AND METHODS: Fifteen participants were divided into two groups. Group 1 (n=10) consisted of patients with signs of unilateral infectious keratitis, while group 2 (n=5) served as the control group. A novel microbiological method was employed to isolate pure cultures of microorganisms. This method involved cultivating microorganisms at two temperature regimes (37 °C and 24 °C) and subsequently identifying them using biochemical, immunological, and physicochemical techniques, including mass spectrometry. Scanning electron microscopy (SEM) with lanthanide staining used as the reference method. The temperature status of the ocular surface was assessed using non-contact infrared thermography. RESULTS: The study demonstrated the presence of psychrotolerant microorganisms on the ocular surface, which exhibited growth at a relatively low temperature of 24 °C. These psychrotolerant microorganisms were found to be isolated from the ocular surface displaying signs of temperature dysregulation. Among such microorganisms are Acinetobacter lwoffii, Achromobacter xylosoxidans, Bacillus licheniformis, Enterococcus faecalis, Klebsiella oxytoca, Klebsiella pneumoniae, Micrococcus luteus, Pseudomonas luteola, Streptococcus spp. CONCLUSION: When identifying the causative agent of infectious keratitis, it is crucial to consider the divergence of growth temperature of ocular surface microorganisms. The presence of psychrotolerant microorganisms on the ocular surface, which can effectively grow at room temperature, should be taken into account, especially in cases of temperature dysregulation.

[Age prediction based on the examination of bone tissue elemental composition using energy dispersive X-Ray spectroscopy]. / Prognozirovanie vozrasta na osnovanii izucheniya elementnogo sostava kostnoi tkani s pomoshch'yu energodispersionnoi rentgenovskoi spektroskopii.

THE AIM OF THE STUDY: Was to assess the possibility of using chemical analysis of bone mineral content by the means of an energy dispersive X-Ray spectroscopy to determine the age of unidentified corpses for forensic identification. A semi-quantative chemical microanalysis of bone fragments of 85 male and female corpses aged between 21 and 91 was done through the use of energy dispersive X-Ray spectrometer. The association of bone tissue apatite mineral composition with age is confirmed and a formula, connecting age and chemical composition change, is proposed. The possibility of using quantitive evaluation of chemical elements content in the normative mineral to determine the unidentified corpse's age in a standard laboratory, equipped with an energy dispersive X-Ray spectrometer, was proved.

[Express-diagnosis of the diseases of the eye and its appendages by scanning electron microscopy with supravital staining]. / Skaniruyushchaya elektronnaya mikroskopiya s supravital'nym kontrastirovaniem v ekspress-diagnostike zabolevanii glaza i pridatochnogo apparata.

This review describes the history of development of a new line of chemical reagents that prompts to significantly reevaluate the application of scanning electron microscopy (SEM) in medical and biological studies, particularly in ophthalmology; considers the establishing of SEM as an analytical method; covers the problems in its application associated with the needs of clinical medicine and the complexities of biological sample preparation for electron microscopy. The article also presents in chronological order the technical solutions associated with creating a unique line of reagents for supravital staining. The multitude of technical solutions allows considering SEM as a method of express diagnostics. The review discusses examples of practical application of these methods for solving certain cases in clinical ophthalmology. The niche of SEM is considered among other methods of clinical diagnostics, as well as its future development involving the use of artificial intelligence.

[Features of accumulation of chemical elements in the volume of the lens in senile cataract]. / Osobennosti nakopleniya khimicheskikh elementov v ob"eme khrustalika pri senil'noi katarakte.

PURPOSE: This study provides a detailed analysis of the bioinorganic chemical composition of lens substance in patients with senile cataract using classical and spatial statistics methods. MATERIAL AND METHODS: The study included 30 isolated human lenses. The light scattering ability (LSA) of the lens substance was evaluated using an original method. Additionally, distribution of chemical elements in the lens substance was analyzed using a scanning electron microscope with energy dispersive spectrometer (SEM/EDS). Measurements by all methods were carried out in a single coordinate space, which made it possible to compare the spatial correlation of different parameters. RESULTS: Small-angle light scattering of the lens substance has been quantitatively characterized for the first time. In contrast to the conventional norm, in senile cataract the accumulation fields of the majority of ion-forming elements (including Na, P, K, Cl) are distributed along the lines repeating the geometry of the lens capsule. At the same time, the light scattering ability of certain areas of the lens is significantly correlated with changes in the concentrations of Na, P, K, Ca in these areas. In particular, one ion-forming element can be distinguished - Na: spatial change of its concentration in senile cataract is strongly associated with a local change in LSA of the lens with opacities clustering of any degree. Thus, a change in the nature of the Na accumulation in the lens volume can be considered the main marker of senile cataract formation. CONCLUSION: The distribution pattern of ion-forming elements indicates that the loss of barrier properties in the capsule plays a significant role in the development of senile cataract.

[Visualization of normal ocular surface microflora via impression cytology sample using scanning electron microscopy with lanthanide contrasting]. / Vizualizatsiya normal'noi mikroflory glaznoi poverkhnosti posredstvom impressionnoi proby s ispol'zovaniem skaniruyushchego elektronnogo mikroskopa i lantanoidnogo kontrastirovaniya.

PURPOSE: To determine the possibilities of impression cytology (IC) with subsequent visualization of the sample on a scanning electron microscope in assessment of normal microflora of the ocular surface. MATERIALS AND METHODS: The article presents a visual characteristic of the microorganisms of the ocular surface (OS) captured during impression cytology (IC) in individuals without signs of inflammatory and degenerative eye diseases. The original method of staining the sample with heavy metal salts made it possible to identify the individual signs of the microorganisms in their subsequent visualization by scanning electron microscopy (SEM). RESULTS: The paper presents photomicrographs of the microorganisms most common for the OS obtained with the help of SEM, confirming and supplementing the data of non-visual methods of studying the ocular microflora. It was shown that the detection frequency of the microbial component of the OS by the visual method presented in this study is comparable with the detection frequency when using the microbial cultivation method (<80%). Coccoid and rod-shaped microorganisms were detected with relatively equal frequency, with the coccoid organisms mainly represented in association with epithelial cells. The morphological diversity of rod-shaped microorganisms is shown. CONCLUSION: The results of the study can be used as a visual reference for the normal microbiome of the eye.

Age-related changes in cationic compositions of human cranial base bone apatite measured by X-ray energy dispersive spectroscopy (EDS) coupled with scanning electron microscope (SEM).

One of the most common scientific methods to study the chemical composition of bone matter is energy-dispersive X-ray spectroscopy (EDS). However, interpretation of the data obtained can be quite complicated and require a thorough understanding of bone structure. This is especially important when evaluating subtle changes of chemical composition, including the age-related ones. The aim of current study is to create a method of processing the obtained data that can be utilized in clinical medicine and use it to evaluate the age evolution of bone chemical composition. To achieve this goal, an elemental composition of 62 samples of cadaver compact bone, taken from the skull base (age: Me = 57.5; 21/91(min/max); Q1 = 39.5, Q3 = 73.75), was studied with EDS. We used the original method to estimate the amount of Mg2+ cations. We detected and confirmed an increase of Mg2+ cation formula amount in the bone apatite, which characterizes age-related resorption rate. Analysis of cation estimated ratio in a normative bone hydroxylapatite showed an increase of Mg2+ amount (R = 0.43, p = 0.0005). Also, Ca weight fraction was shown to decrease with age (R = - 0.43, p = 0.0005), which in turn confirmed the age-dependent bone decalcification. In addition, electron probe microanalysis (EPMA) and X-ray diffraction analysis (XRD) were performed. EDS data confirmed the EPMA results (R = 0.76, p = 0.001). In conclusion, the proposed method can be used in forensic medicine and provide additional data to the known trends of decalcification and change of density and crystallinity of mineral bone matter.

Moisture adsorption by decellularized bovine pericardium collagen matrices studied by terahertz pulsed spectroscopy and solid immersion microscopy.

In this paper, terahertz (THz) pulsed spectroscopy and solid immersion microscopy were applied to study interactions between water vapor and tissue scaffolds-the decellularized bovine pericardium (DBP) collagen matrices, in intact form, cross-linked with the glutaraldehyde or treated by plasma. The water-absorbing properties of biomaterials are prognostic for future cell-mediated reactions of the recipient tissue with the scaffold. Complex dielectric permittivity of DBPs was measured in the 0.4-2.0 THz frequency range, while the samples were first dehydrated and then exposed to water vapor atmosphere with 80.0 ± 5.0% relative humidity. These THz dielectric measurements of DBPs and the results of their weighting allowed to estimate the adsorption time constants, an increase of tissue mass, as well as dispersion of these parameters. During the adsorption process, changes in the DBPs' dielectric permittivity feature an exponential character, with the typical time constant of =8-10 min, the transient process saturation at =30 min, and the tissue mass improvement by =1-3%. No statistically-relevant differences between the measured properties of the intact and treated DBPs were observed. Then, contact angles of wettability were measured for the considered DBPs using a recumbent drop method, while the observed results showed that treatments of DBP somewhat affects their surface energies, polarity, and hydrophilicity. Thus, our studies revealed that glutaraldehyde and plasma treatment overall impact the DBP-water interactions, but the resultant effects appear to be quite complex and comparable to the natural variability of the tissue properties. Such a variability was attributed to the natural heterogeneity of tissues, which was confirmed by the THz microscopy data. Our findings are important for further optimization of the scaffolds' preparation and treatment technologies. They pave the way for THz technology use as a non-invasive diagnosis tool in tissue engineering and regenerative medicine.

Biomineralization of Trabecular Meshwork in Open-Angle Glaucoma.

A new form of open-angle glaucoma has been identified, in which calcification and silicification of the trabecular meshwork is a potentially significant component of outflow obstruction. It is noted that the mineralization of this area is promoted by various disturbances in the acid-base balance in the tissue. The role of melanosomal enzymes in the initiation of the formation of mineral calcium phosphate in trabecular tissue in open-angle glaucoma is considered.

[Accumulation of sulfur and phosphorus in the eye's drainage system in primary open-angle glaucoma]. / Nakoplenie sery i fosfora v drenazhnoi zone glaza pri pervichnoi otkrytougol'noi glaukome.

PURPOSE: To analyze the chemical composition of the sclera and trabecular meshwork in patients with primary open-angle glaucoma (POAG) and to assess the impact of corneal-compensated intraocular pressure (IOP) on the chemistry of the drainage area. MATERIAL AND METHODS: Biopsy specimens of the trabecular meshwork (89 specimens) and sclera (41 specimens) obtained from patients with POAG on maximal medical therapy were analyzed to determine the content of certain chemical elements - carbon (C), nitrogen (N), oxygen (O), aluminum (Al), calcium (Ca), chlorine (Cl), potassium (K), magnesium (Mg), sodium (Na), phosphorus (P), silicon (Si) and sulfur (S). The elements were selected based on chemical structure target tissue and sensitivity of the method used for analysis. Visualization was performed using the «EVO LS 10¼ (Zeiss, Germany) scanning electron microscope (SEM), and the chemical composition was studied with the «Oxford-X-MAX-50¼ (Oxford, UK) energy-dispersive spectrometer (EDS). Statistical analysis was performed using the Spearman correlation. RESULTS: The IOP of patients with POAG was found to have positive correlation with the total amount of S, P, and Ca in the trabecular meshwork. The accumulation of sulfur-containing compounds associated with pigment granules in trabecular meshwork's tissue was identified. A correlation was determined between the organic and inorganic (mineral) phosphorus content in the trabeculae, and the IOP values. The organic component of phosphorus was better represented than the mineral component in patients with increased IOP. CONCLUSION: In patients with POAG, an increase in the IOP level causes the amount of S associated with pigment granules and the proportion of organic P to increase in the trabecular meshwork, which should be taken into account in the further search for drug therapy that would potentially affect pathologically altered tissue.

Bioinorganic chemistry of open-angle glaucoma: A review.

At present, physical methods of chemical analysis are constantly improving providing large amount of data on elemental composition of organs and tissues. However, only few works describe the correlation (or the potential connection) between the general or local bioelemental imbalances and specific biochemical reactions that are involved in pathogenesis of certain diseases. This review describes primary open-angle glaucoma (POAG) - one of the most common ophthalmic diseases - in terms of elemental chemistry. The authors look into the impact that various subgroups of elements have on passive and active processes of homeostasis regulation and hydrodynamic balance in the eye. Alkaline metals and their analogues (K, Na, Li, Rb, Cs) influence hydrostatics and hydrodynamics by means of both K-Na pumps and osmosis. Alkaline-earth elements and their analogues (Ca, Mg, Sr, Ba, Be) are involved in biomineralization and intercellular interaction in the drainage areas. Chalcophile metals and their analogues (Zn, Cu, Hg, Co, Ni, Cd, Pb, Mo, Sb) regulate redox reactions. They are the cofactors of enzymes that support structural homeostasis of the drainage area. Siderophile metals (Fe, Mn, Cr, Rh) regulate oxidation-reduction reactions, including those associated with limited nutrition of tissues in glaucoma. The role of amphoteric metals and nonmetals (Al, Si, Ga, V, TI, Sn, Ge, Zr, W) in POAG has not been described properly, but they were noted to participate in mineralization. Structure-forming non-metals and their analogues (N, S, Se, As) are directly involved in the formation of protein and non-protein aggregates that prevent aqueous humor outflow. The specific role of phosphorus in the pathogenesis of glaucoma has not been described previously. The authors analyze the involvement of phosphorus in energy-dependent processes of cellular activity, which are aimed at the reprocessing of aggregates that cause aqueous humor retention.

[Hypotensive efficacy of bimatoprost in monotherapy of primary open-angle glaucoma]. / Gipotenzivnaia éffektivnost' bimatoprosta v monoterapii pervichnoi otkrytougol'noi glaukomy.

PURPOSE: To study the effectiveness of monotherapy with bimatoprost in patients with decompensated intraocular pressure (IOP). MATERIAL AND METHODS: 90 patients (132 eyes) with stage I-II glaucoma and decompensated IOP previously treated with timolol, latanoprost and travoprost were included in the study. Average IOP at the beginning of the study was 25.4±2.5 mmHg. All patients had their hypotensive medications substituted by bimatoprost. The patients were examined during the initial appointment, as well as after 4 and 12 weeks. RESULTS: IOP decrease was highest in the group of patients who had been treated with timolol: initially IOP was 26.2±1.8 mmHg, after 4 weeks - 21.0±2.2 mmHg, after 12 weeks - 20.8±1.9 mmHg (p<0.001). The initial IOP of patients who had been treated with latanoprost was 24.8±2.9 mmHg, 21.8±2.4 mmHg after 4 weeks, and 21.6±2.3 mmHg after 12 weeks (p<0.001). Patients who had been treated with travoprost had 25.6±2.2 mmHg initially, 23.0±2.5 mmHg after 4 weeks, and 23.2±2.6 mmHg after 12 weeks (p<0.001). By the end of the study IOP has decreased by 5.4, 3.2 and 2.4 mmHg in the groups of patients who had been treated with timolol, latanoprost and travoprost, respectively. CONCLUSION: Bimatoprost can be used as monotherapy if another hypotensive drug in monotherapy is insufficient for IOP compensation.