Seasonal Changes in the Antioxidant Activity and Biochemical Parameters of Goat Milk.

Goats are ubiquitous, including in hot and dry regions, while also being very sensitive to climate fluctuations, expressed in temperature differences. This affects their productivity and milk quality. Adaptation to heat requires high energy costs, affects "neurohumoral" regulation and is accompanied by oxidative stress with the increased production of free radicals. The aim was to study the main biochemical parameters of goat milk and its antioxidant activity depending on the season of the year. Sampling was carried out in April, June, August and October. Analysis of the biochemical components and antioxidant activity of goat milk was performed using modern analytical systems. From spring to autumn, the mass fraction of true or crude proteins in goat milk increased by 14.6-63.7% or by 12.3-52.1%, and the mass fraction of caseins also increased by 13.6-60.6%. For vitamin C level and the total amount of water-soluble antioxidants, a pronounced gradual decrease from spring to autumn was observed. In the summer period, a small increase in the carotene level in milk (by 3.0-6.1% compared to April) was established. Vitamin A content increased by 86.5% (June) or by 70.3% (October) compared to April. Thus, the numerous significant changes in the major parameters of goat's milk depending on the season were revealed.

Relationships between the Content of Micro- and Macroelements in Animal Samples and Diseases of Different Etiologies.

Many of the micro- and macro-elements (MMEs) required by the body are found in environmental objects in concentrations different from their original concentration that can lead to dangerous animal diseases ("microelementoses"). The aim was to study the features of MME (accumulating in wild and exotic animals) in connection with particular diseases. The work using 67 mammal species from four Russian zoological institutions was completed in 2022. Studies of 820 cleaned and defatted samples (hair, fur, etc.) after "wet-acid-ashing" on an electric stove and in a muffle furnace were performed using a Kvant-2A atomic absorption spectrometer. The content of zinc, copper, iron, cadmium, lead, and arsenic was assessed. The level of MME accumulation in the animal body contributes not only to the MME status and the development of various concomitant diseases, but the condition itself can occur by intake of a number of micronutrients and/or drugs. Particular correlations between the accumulation of Zn and skin, oncological diseases, Cu-musculoskeletal, cardiovascular diseases, Fe-oncological diseases, Pb-metabolic, nervous, oncological diseases, and Cd-cardiovascular diseases were established. Therefore, monitoring of the MME status of the organism must be carried out regularly (optimally once every 6 months).

Correlations between the Major Amino Acids and Biochemical Blood Parameters of Pigs at Controlled Fattening Duration.

Analytical control of protein and amino acid (AA) contents of animal tissues is an important problem in the fundamental and applied aspects. The aims of the work were the following: to measure the pig blood AAs; and to establish the correlations between AAs and biochemical parameters in dependence on the pig fattening duration. All 80 animals were divided onto 4 animal groups: 65, 72, 82, and 90 fattening days. The correlations between AAs and the total protein or its fractions (TP&F), nitrogen metabolites, carbohydrates, lipids, some enzymes in the pig blood for each of these animal groups obtained for the first time. The authors established the following total amounts of correlation coefficients (with reasonable p-values) in each of the group separately: group 1, 1* (p < 0.05); group 2, 0; group 3, 28* (p < 0.05) and 9** (p < 0.01); group 4, 28* (p < 0.05) and 25** (p < 0.01). Thus, about 82−90 days (groups 3 and 4) can be the optimal for the pig fattening, based on the correlation analysis for the numerous data of major AA and biochemical parameters of pig blood. These results can be useful for animal health monitoring and husbandry.

Correlations between the Total Antioxidant Activity and Biochemical Parameters of Cow Milk Depending on the Number of Somatic Cells.

The aim of the work was to study the correlations between the total amount of water-soluble antioxidants (TAWSA) and biochemical parameters (BC) of cow milk depending on the somatic cell count (SCC). The BC and TAWSA values of cow milk were measured by spectroscopic and amperometric methods, respectively. The milk samples from the black-and-white cows (Moscow region) were divided according to SCС values: (1) ≤200, (2) 200-499, (3) 500-999, and (4) ≥1000 thousand units/mL. The average TAWSA values for groups 1, 2, 3, and 4 (33, 15, 13, and 12 milk samples) were the following: 15.95 ± 0.74, 14.45 ± 0.84, 16.04 ± 0.63, and 14.58 ± 1.18. The correlations between TAWSA and BC (group 1) were the following: total fat percentage (TFP) -0.305; true protein percentage (TP1) -0.197; total nitrogen percentage (TN2) -0.210; lactose -0.156; solids-not-fat (SNF) -0.276; total dry matter (TDM) -0.399; freezing point (FP) -0.112; pH -0.114; somatic cell count (SCC) - (-0,052). The correlations between TAWSA and BC (group 2) were the following: TFP -0.332; TP1 -0.296; TN2 -0.303; lactose - (-0.308); SNF -0.159; TDM -0.391; FP -0.226; pH - (-0.211); SCC -0.193. The correlations between TAWSA and BC (group 3) were the following: TFP - (-0.352); TP1 - (-0.411); TN2 - (-0.401); lactose - (-0.166); SNF - (-0.462); TDM - (-0.504); FP - (-0.766); pH - (-0.047); SCC - (-0.698). The correlations between TAWSA and BC (group 4) were the following: TFP -0.159; TP1 -0.046; TN2 - 0.077; lactose - (-0.317); SNF - (-0.237); TDM -0.058; FP - (-0.036); pH - (-0.477); SCC - (-0.072). These data are important in assessing the physiological-biochemical status and state of the antioxidant defense system of cows' organism.

Correlations between Antioxidant and Biochemical Parameters of Blood Serum of Duroc Breed Pigs.

Correlations between the major biochemical (BC) and antioxidant (TAWSA) parameters of pigs' blood are necessary to study in order to assess physiological-biochemical status (PhBS), animal health, production, etc. Blood samples were obtained from Duroc breed boars (n = 77), divided into groups 1 (n = 25), 2 (n = 40) and 3 (n = 12), which were fattened for 65, 72 and 100 days, respectively. Significant positive and negative correlations were found between TAWSA and BC parameters of pigs' blood for group 3: very high in the case of total protein (TP) (-0.75) and aspartate aminotransferase (AST) (-0.79); high in the case of cholesterol (-0.72), glucose (0.66), alkaline phosphatase (0.66), calcium ions (-0.60) and globulins (0.53); moderate in the case of albumins (-0.36), triglycerides (-0.35), magnesium (-0.32) and phosphorus (-0.27). The same was found for group 2: high in the case of TP (0.51); moderate in the case of globulins (0.48), cholesterol (0.33) and phosphates (0.25). The only moderate correlation was found for group 1: magnesium (-0.48), glucose (0.36) and calcium (-0.25). This tendency indicated the stabilization of pig PhBS during growth and fattening, which can be useful for understanding the PhBS and antioxidant features of pigs, the factors of their nutrition, maintenance, etc.

Biochemical parameters, dynamic tensiometry and circulating nucleic acids for cattle blood analysis: a review.

The animal's blood is the most complicated and important biological liquid for veterinary medicine. In addition to standard methods that are always in use, recent technologies such as dynamic tensiometry (DT) of blood serum and PCR analysis of particular markers are in progress. The standard and modern biochemical tests are commonly used for general screening and, finally, complete diagnosis of animal health. Interpretation of major biochemical parameters is similar across animal species, but there are a few peculiarities in each case, especially well-known for cattle. The following directions are discussed here: hematological indicators; "total protein" and its fractions; some enzymes; major low-molecular metabolites (glucose, lipids, bilirubin, etc.); cations and anions. As example, the numerous correlations between DT data and biochemical parameters of cattle serum have been obtained and discussed. Changes in the cell-free nucleic acids (cfDNA) circulating in the blood have been studied and analyzed in a variety of conditions; for example, pregnancy, infectious and chronic diseases, and cancer. CfDNA can easily be detected using standard molecular biological techniques like DNA amplification and next-generation sequencing. The application of digital PCR even allows exact quantification of copy number variations which are for example important in prenatal diagnosis of chromosomal aberrations.

Biochemical aspects of lipase immobilization at polysaccharides for biotechnology.

The design of immobilized enzyme preparations is an important and relevant area of modern sciences and technologies. Immobilization of enzymes from animal sources (component I) on natural carriers (component II) increases the system stability by protecting the active site of the enzyme from deactivation; facilitates the separation and accelerates the recovery of the enzyme. This makes reuse possible and provides a significant reduction in operating costs. Hydrolytic enzymes (such as lipases) and polysaccharides (such as chitosan) are the most promising of such pairs of components. The main attention here is devoted to the discussion on lipase immobilization on polysaccharide (mainly - chitin and chitosan). Based on the analysis of the available literature, the most adequate method is the immobilization of lipase from porcine pancreas (LPP) on polysaccharide particles (such as chitin or chitosan) pre-treated with ultrasound (to increase the particle surface area) and glutaraldehyde (for particle activation) that shows reasonably high LPP activity and stability. In order to increase further the activity of the lipase, some authors proposed to incorporate a spacer in the form of 1,3-diaminopropane (or 1,3-diaminobutane) prior to activation of the surface of the chitosan particles. In particular cases, the use of chitin (instead of chitosan) may be an alternative solution for biotechnological applications. Recently the idea of constructing "supramolecular enzyme systems" realized in the so-called "coimmobilized multienzymatic systems" strategy. The most fascinating example is the combined assay of a mixture of native LPP, glycerol kinase (from Cellulomonas) and glycerol-3-phosphate oxidase (from Aerococcus viridans) linked by glutaraldehyde to chitosan (as shell for inorganic nanoparticle core). This material was placed on a Pt-electrode as biosensor and was successfully applied for amperometric determination of the triglyceride level in the serum of healthy and diseased person. Thus, the whole innovative research-production sequence is described by Aggarwal V. and Pundir C.S.: from simple components to advanced material and further biomedical application. Thus, the following approach of lipase immobilization appears the most promising for future applications: a few types of lipases or the combination of LPP with some other enzymes immobilized simultaneously on multifunctional carriers (as nanohybrids of inorganic core and polysaccharide shell).

Cell Staining by Photo-activated Dye and Its Conjugate with Chitosan.

Photo-activated or "Caged" rhodamine dyes are the most useful for microscopic investigation of biological tissue by various fluorescent techniques. Novel precursor of the fluorescent dye (PFD813) has been studied for photosensitive staining of numerous animal cells. The functional rhodamine dye (Rho813) with intensive fluorescence has been obtained after photoactivation of its precursor PFD813 inside cells. The dye Rho813 has been successfully used for the optical detection of particular features in biological objects (HaCaT cells, HBL-100, MDCK, lymphocytes). Moreover, the chitosan conjugate with PFD molecules ("Chitosan-PFD813″) has been obtained and studied for the first time. The developed procedures and obtained data are important for further applications of novel precursors of fluorescent dyes ("caged" dyes) for microscopic probing of biological objects. As example, the synthesized "Chitosan-PFD813″ has been successfully applied in this study for intracellular transport visualization by fluorescent microscopy.

Multifunctional membranes based on photosensitive crown-ether derivatives with advanced properties.

This review discusses recent works on monolayer, multilayer and polymer films of various crown-ether derivatives. Preparation and investigation of such membrane nanostructures based on photosensitive and surface-active crown-ethers is a rapidly growing field at the "junction" of colloids and polymers, materials sciences and nanotechnology. These membranes can serve as convenient models for studying the self-organization and molecular recognition processes at interfaces that are typical for biomembranes. The results obtained for such structures by absorption and fluorescence spectroscopy, atomic force and Brewster-angle microscopy, surface pressure and surface potential isotherm measurements have been described. The possibility of developing multifunctional materials possessing advanced properties has been demonstrated.

Conjugates of a photoactivated rhodamine with biopolymers for cell staining.

Conjugates of the photoactivated rhodamine dyes with biopolymers (proteins, polysaccharides, and nucleic acids) are important tools for microscopic investigation of biological tissue. In this study, a precursor of the photoactivated fluorescent dye (PFD) has been successfully used for staining of numerous mammalian cells lines and for conjugate formation with chitosan ("Chitosan-PFD") and histone H1 ("Histone H1.3-PFD"). The intensive fluorescence has been observed after photoactivation of these conjugates inside cells (A431, HaCaT, HEK239, HBL-100, and MDCK). Developed procedures and obtained data are important for further application of novel precursors of fluorescent dyes ("caged" dyes) for microscopic probing of biological objects. Thus, the synthesized "Chitosan-PFD" and "Histone H1-PFD" have been successfully applied in this study for intracellular transport visualization by fluorescent microscopy.

Controlled influence of quantum dots on purple membranes at interfaces.

The development of bio-sensitized nanofilms engineered from biomembrane components and inorganic nanoparticles is a promising field of colloid and interface science and technologies. Recent nano-bioengineering approaches employing quantum dots (QDs) permit the enhancement of the purple membrane (PM) "light-harvesting capacity" compared to native PMs. The influence of QDs on the PM properties, especially the bacteriorhodopsin (bR) photocycle, has been found that has both fundamental (mechanisms of photoreception) and applied implications (including the fabrication of hybrid bionanomaterials). Samples of PM-QD complexes capable of energy transfer and characterized by increased rates of M-intermediate formation and decay have been obtained. The modified bR photocycle kinetic parameters may be explained by changes in the PM interface upon QD adsorption. The increase and decrease in absorption at 410 nm (or photopotential) for PM-QD complexes are, on average, several times more rapid than for PM suspensions or PM dry films. These results provide a strong impetus for the development of nanomaterials with advanced properties.

Novel precursors of fluorescent dyes. 1. Interaction of the dyes with model phospholipid in monolayers.

Photoactivated ("caged") fluorescent dyes are modern tools for structure and function studies of cell membranes and subcellular organelles. Recently synthesized precursors of rhodamine fluorescent dyes (abbreviations PFD813 and PFD814) important for microscopic probing of biological objects have been studied in solution. In order to characterize the behavior at interfaces, monolayers of PFD813 and PFD814 on water have been formed and investigated. The interactions of these precursors with the biomembrane component dimyristoylphosphatidylethanolamine in monolayers at the air-water interface and after transfer to glass plates have been studied by measuring monolayer parameters and spectroscopic properties before and after photo-chemical formation of the fluorescent rhodamine dyes Rho813 and Rho814, respectively.

Thin films and assemblies of photosensitive membrane proteins and colloidal nanocrystals for engineering of hybrid materials with advanced properties.

The development and study of nano-bio hybrid materials engineered from membrane proteins (the key functional elements of various biomembranes) and nanoheterostructures (inorganic colloidal nanoparticles, transparent electrodes, and films) is a rapidly growing field at the interface of materials and life sciences. The mainspring of the development of bioinspired materials and devices is the fact that biological evolution has solved many problems similar to those that humans are attempting to solve in the field of light-harvesting and energy-transferring inorganic compounds. Along this way, bioelectronics and biophotonics have shown considerable promise. A number of proteins have been explored in terms of bioelectronic device applications, but bacteriorhodopsin (bR, a photosensitive membrane protein from purple membranes of the bacterium Halobacterium salinarum) and bacterial photosynthetic reaction centres have received the most attention. The energy harvesting in plants has a maximum efficiency of 5%, whereas bR, in the absence of a specific light-harvesting system, allows bacteria to utilize only 0.1-0.5% of the solar light. Recent nano-bioengineering approaches employing colloidal semiconductor and metal nanoparticles conjugated with biosystems permit the enhancement of the light-harvesting capacity of photosensitive proteins, thus providing a strong impetus to protein-based device optimisation. Fabrication of ultrathin and highly oriented films from biological membranes and photosensitive proteins is the key task for prospective bioelectronic and biophotonic applications. In this review, the main advances in techniques of preparation of such films are analyzed. Comparison of the techniques for obtaining thin films leads to the conclusion that the homogeneity and orientation of biomembrane fragments or proteins in these films depend on the method of their fabrication and increase in the following order: electrophoretic sedimentation < Langmuir-Blodgett and Langmuir-Schaefer methods < self-assembly and layer-by-layer methods. The key advances in the techniques of preparation of the assemblies or complexes of colloidal nanocrystals with bR, purple membranes, or photosynthetic reaction centres are also reviewed. Approaches to the fabrication of the prototype photosensitive nano-bio hybrid materials with advanced photovoltaic, energy transfer, and optical switching properties and future prospects in this field are analyzed in the concluding part of the review.

Membrane and Films Based on Novel Crown-Containing Dyes as Promising Chemosensoring Materials.

This paper discusses several works on supramolecular systems such as monolayer and multilayer, polymer films of various crown-containing dyes, surface-active monomers and polymers. Design, production and investigation of the membrane nanostructures based on crown ethers is a rapidly developing field at the "junction" of materials sciences and nanotechnology. These nanostructures can serve as convenient models for studying the self-organization and molecular recognition processes at interfaces that are typical for biomembranes. Based on the results obtained for such structures by absorption and fluorescence spectroscopy, atomic force and Brewster-angle microscopy, surface pressure and surface potential isotherm measurements, the possibility of developing micro- and nanomaterials possessing a set of specified properties (including chemosensor, photochromic and photorefractive materials) is demonstrated.

Influence of monolayer state on spectroscopy and photoisomerization of an amphiphilic styryl-pyridinium dye on a solid substrate.

The spectroscopy and photochromic properties of transferred monolayers of the amphiphilic styryl-pyridinium dye 4-(3',4'-dimethoxystyryl)-N-octadecylpyridinium perchlorate (DMPOP) were studied at different conditions during their transfer. The emission maxima of the monolayers transferred from the air-water interface in the liquid-expanded phase are strongly dependent on the surface pressure applied during the transfer process, even at values when the area per molecule is 2-3 times larger than the area occupied by a chromophore. In monolayers transferred from the liquid-condensed phase, the presence of a different kind of aggregates was observed. The fluorescence emission properties of the monolayers can be reversibly modulated by photoinduced E-Z isomerization. A blue shift up to 72 nm in the emission maximum, depending on the transfer conditions of the films, can be obtained by irradiation with blue light, and partially recovered (a red shift of up to 26 nm) with UV radiation. The rate at which the first process (E-->Z) takes place is drastically reduced in monolayers transferred from the liquid-condensed phase as compared to those transferred from the liquid-expanded one. However, the rate of the reverse reaction (Z-->E) is not significantly altered. These properties make DMPOP a promising material for the preparation of Langmuir-Blodgett films, whose properties can be effectively controlled by the transfer conditions and subsequently optically modulated, for potential applications as photonics devices for data storage.