Whole Blood Content Research Articles

Advancing towards practice: A novel LC-MS/MS method for detecting retinol in dried blood spots

BackgroundTo date, clinical laboratories face challenges in quantifying retinol from DBS samples. Disputes arise throughout the whole detection process, encompassing the storage condition, the release strategy as well as the selection of internal standards. MethodsWe incubated DBS with ascorbic acid solution. Then, retinol-d4 in acetonitrile was introduced to incorporate isotopic internal standard and promote protein precipitation. Afterward, sodium carbonate solution was added to ionize cytochromes (such as bilirubin), which amplified the difference of their hydrophobicity to retinol. Subsequently, cold-induced phase separation could be facilitated to separate retinol from the impurities. In the end, the upper layer was injected for LC-MS/MS analysis. ResultsBy comparing the detected retinol content in whole blood and DBS samples prepared from the same volume, we confirmed the established pretreatment was capable to extract most of retinol from DBS (recovery >90 %). Thereafter, we verified that within DBS, retinol possessed satisfying stability without antioxidation. Indoor-light exposure and storage duration would not cause obvious degradation (<10 %). Following systematic validation, the established method well met the criteria outlined in the relevant guidelines. After comparing with detected DBS results to the paired plasma samples, 54 out of 60 met the acceptance limit for cross-validation of ±20 %. ConclusionsWe realized precise quantification of retinol from one 3.2 mm DBS disc. By circumventing conventional antioxidation, liquid-liquid/solid-phase extraction and organic solvent evaporation, the pretreatment could be completed within 15 min consuming only minimal amounts of low-toxicity chemicals (ascorbic acid, acetonitrile, and sodium carbonate). We expect this contribution holds the potential to significantly facilitate the evaluation of patients' vitamin A status by using DBS samples in the future.

Acute Cannabis Administration Transiently Reduces Mitochondrial DNA in Young Adults: Findings from a Secondary Analysis of a Double-Blind, Placebo-Controlled, Randomized Clinical Trial.

Background: Cannabis is one of the world's most commonly used substances; however, many questions remain unanswered as to how cannabis impacts the body. Recently, there has been a resurgence of research into the effects of plant-derived cannabinoids on mitochondrial health. In particular, a number of studies implicate mitochondrial-Δ9-tetrahydrocannabinol (Δ9-THC) interactions with altered memory, metabolism, and catalepsy in mice. Although the research in this field is expanding rapidly, there is little known about the effects of cannabis on mitochondria health in human subjects either in acute or chronic term use. Methods: Blood samples were obtained from a double-blind, placebo-controlled, parallel-group randomized clinical trial in which adults who regularly use cannabis (1-4 days/week) aged 19-25 years were randomized 2:1 to receive either an active (12.5% Δ9-THC) cigarette or placebo (<0.01% Δ9-THC) cigarette containing 750 mg of cannabis before driving simulator testing. DNA was extracted from whole blood using commercial spin columns, followed by measurement of mt-ND1, mt-ND4, and β2M using quantitative polymerase chain reaction. One-way repeated measures analysis of variance (ANOVA) followed by Dunnett's multiple comparisons test was used to observe changes in mitochondrial DNA (mtDNA) copy number over time. A two-tailed Pearsons R test was used to assess correlations between mtDNA copy number and cannabinoid levels (Δ9-THC and metabolites) in blood. Results: We found that exposure to active cannabis containing Δ9-THC, as opposed to placebo, was associated with an acute reduction in mitochondrial DNA copy number in whole blood at 15 min and 1 h after smoking. The observed decrease in mtDNA copy number negatively correlated with blood concentrations of 11-hydroxy-Δ9-tetrahydrocannabinol (11-OH-THC) and 11-Nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH), the two primary metabolites of Δ9-THC, but not Δ9-THC itself. Further, the negative correlation between 11-OH THC and THC-COOH concentrations and mtDNA copy number was found in only a subgroup of participants who use cannabis infrequently, suggesting a tolerance effect. Conclusions: These results illuminate mitochondrial alterations attributed to Δ9-THC consumption, which may be mediated by metabolites. These results appear to suggest stronger effects in individuals who consume cannabis less frequently, suggesting some form of tolerance to the effects of Δ9-THC and its metabolites on mtDNA content in whole blood. Keywords: Mitochondria; mtDNA; cannabis; THC; THC metabolites; blood; THC-COOH; 11-OH-THC.

The influence of serotonin on the formation of clinical symptoms of functional gastrointestinal disorders in children

According to the Rome IV consensus (2016), functional gastrointestinal disorders (FGIDs) are disorders of the interaction between the gut and the brain. A crucial role in the realization of this mechanism is played by the key neurotransmitter serotonin. Aim - to investigate the influence of serotonin on the formation of clinical symptoms in children with FGIDs. Materials and methods. Under observation were 72 children aged 10-18 years with FGIDs. The Group I consisted of 30 children with combined functional dyspepsia (FD) and irritable bowel syndrome (IBS), the Group II included 42 children with FD. A control group of 20 healthy children was established. The diagnosis of FD and IBS was established based on the analysis of clinical symptoms according to the Rome IV criteria (2016). The severity of clinical symptoms was assessed using the Likert scale from 0 to 5 points. The study of serotonin content in whole blood was conducted using a fluorometric method after adsorption on carboxymethylcellulose. Results. The intensity of abdominal pain and the severity of dyspeptic syndrome in children with combined FD and IBS is higher than in children with FD (p&lt;0.01; p&lt;0.05, respectively). The serotonin content in whole blood in children with FGIDs is lower than in healthy children (p&lt;0.001). In children with combined FD and IBS, the serotonin content is lower than in children with FD (p&lt;0.05). Reverse correlation links were found between serotonin content and the intensity of pain syndrome (r=-0.7), the severity of dyspeptic syndromes (r=-0.5), constipation (r=-0.7), and diarrhea (r=0.7). Conclusions. The obtained research results indicate the influence of serotonin on the formation of clinical symptoms in children with FD and combined FD and IBS. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.

Multidimensional analysis of the essential elements in pregnant women's whole blood and characterization of maternal status by elemental pattern

BackgroundDuring pregnancy, the fetus needs to obtain a lot of nutrients from the mother, but the micronutrient deficiencies in pregnancy are not clear at present, and there is no reliable basis for nutrient intake and supplement. The purpose of this study was to understand the levels of essential elements in whole blood of pregnant women during various pregnancy stages at different ages and in different regions, to evaluate the deficiency of essential elements in Chinese pregnant women, and to explore the feasibility of using the elemental pattern to characterize maternal status. MethodsWhole blood samples of 11222 healthy pregnant women enrolled in different areas of China from Jan–Dec 2019, were analyzed for concentrations of six essential elements including Mn, Cu, Zn, Ca, Mg, and Fe, using the inductively coupled plasma mass spectrometer. A retrospective comparative study during different pregnancy periods at different ages and in different regions in whole blood essential elements content from non-pregnant normal women and pregnant normal women was developed using multivariate statistical analysis. Principal component analysis evaluation elemental pattern was used to characterize pregnancy status of pregnant women. ResultsIn general, the levels of six essential elements in whole blood of pregnant women can satisfy the needs of normal physiological activities. With the development of pregnancy, the contents of Cu and Mn increased, while the contents of Fe and Mg decreased, and the contents of Zn and Ca have no noteworthy change. At the same gestation stage, the Cu content in whole blood of elderly pregnant women was higher. There were some differences in whole blood essential elements content of pregnant women in different regions. Principal component analysis and heat map analysis showed the feasibility of using bioinformatics research strategies to identify different pregnancies. ConclusionsThere are differences in the content of whole blood essential elements of women at different stages of pregnancy in different regions. It was found that there was no obvious deficiency in whole blood essential elements levels of pregnant women in recent years. The pattern of essential elements has a certain application potential in the evaluation of pregnancy and pregnant women's health status.

The Role of SOCS2 Cytokine Signaling Suppressor in the Regulation of ProInflammatory Activity of Whole Blood Cells after Lower Respiratory Tract Infection

The aim of the investigation was to study the relationship between the content of whole blood in mononuclear leukocytes in pneumonia and in apparently healthy individuals of cytokine signaling suppressor 2 (SOCS2) with the production of cytokines (TNFα, TGFb, IFNα, IFNβ, IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-10, IL-12, IL-17A, RAIL-1, RANTES) and individual factors of the NF-kB and JAK / STAT signaling pathways (NF-kB2, p65, p50, STAT1, STAT3, STAT5B, STAT6). Materials and research methods . The research material was mononuclear cells isolated from venous blood samples, as well as blood plasma of practically healthy individuals and patients with pneumonia. In nuclear-cytoplasmic lysates of mononuclear blood cells, the concentration of the components of the nuclear transcription factor NF-κB, p65, p50, NF-κB2, factors STAT1, STAT3, STAT5B, STAT6, and protein SOCS2, was assessed by enzyme immunoassay. We also determined the concentration of TNFα, IL-1β, TGFb, IFNα, IFNβ, IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-10, IL-17A, RAIL-1, RANTES. The results of this study indicate that the stage of pneumonia convalescence is accompanied by dysregulation of the production of the main proinflammatory cytokines, manifested by a decrease in the level of TNFα, TGFb, RANTES, IL-4, IL-17A, IFNβ, IFNγ and an increase in the production of IL-2 and IFNα. Against this background, a decrease in the phosphorylation of the STAT3 and STAT4 factors was noted, as well as a decrease in the content of p50 and p65 proteins in MNCs. These changes were associated with an increased content of the SOCS2 factor in MNCs. The analysis showed that an increase in the content of SOCS2 in MNCs from the minimum level determined by the concentration corresponding to the 1st quartile of the sample (1.3 ng / ml) to the maximum, determined by the 4th quartile of the sample (1.7 ng / ml) is associated with a decrease in production IL-1β, IL-4, IL-4, IL-5, IL-10, IL-17A, TGFb, RANTES and IFNβ against the background of an increase in the level of INFα, INFγ and IL-2. Changes in cytokine production were accompanied by an increase in STAT5B, STAT4, and NF-kB2 levels and a decrease in STAT3 phosphorylation. a decrease in the content in the cell of the components of the nuclear transcription factor NF-κB, in particular, p50, p65. Conclusion . The peculiarities of the relationship of SOCS2 with the studied factors suggests that its high level helps to limit the production of proinflammatory cytokines, in particular those produced by type 2 T-helpers and Th17, stimulates an increase in ICC sensitivity to IL-2 and stimulation of type 1 T-helpers. These effects are realized due to an increase in the phosphorylation of the STAT5 and STAT4 factors, a decrease in the STAT3 activity, and a change in the ratio of the components p50, p65 and NF-κB2 of the nuclear transcription factor NF-κB in the cell.

Stimulating the expression of sphingosine kinase 1 (SphK1) is beneficial to reduce acrylamide-induced nerve cell damage

Sphingosine kinase 1 (SphK1) is an important signaling molecule for cell proliferation and survival. However, the role of SphK1 in acrylamide (ACR)-induced nerve injury remains unclear. The purpose of this study was to investigate the role and potential mechanism of SphK1 in ACR-induced nerve injury. Liquid chromatography triple quadrupole tandem mass spectrometry (LC-MS/MS) and reverse transcription-quantitative PCR (RT-qPCR) were used to detect sphingosine 1-phosphate (S1P) content in serum and SphK1 content in whole blood from an occupational work group exposed to ACR compared to a non-exposed group. For in vitro experiments, SphK1 in human SH-SY5Y neuroblastoma cells was activated using SphK1-specific activator phorbol 12-myristate 13-acetate (PMA). Our research also utilized cell viability assays, flow cytometry, western blots, RT-qPCR and related protein detection to assess activity of the mitogen activated protein kinase (MAPK) signaling pathway. The results of the population study showed that the contents of SphK1 and S1P in the ACR-exposed occupational contact group were lower than in the non-exposed group. The results of in vitro experiments showed that expression of SphK1 decreased with the increase in ACR concentration. Activating SphK1 improved the survival rate of SH-SY5Y cells and decreased the apoptosis rate. Activating SphK1 in SH-SY5Y cells also regulated MAPK signaling, including enhancing the phosphorylation of extracellular signal-regulated protein kinases (ERK) and inhibiting the phosphorylation of c-Jun N-terminal kinase (JNK) and p38. These results suggest that activating SphK1 can protect against nerve cell damage caused by ACR.

Relationships between lead-induced learning and memory impairments and gut microbiota disturbance in mice

Objective: To explore the effect of lead exposure on the neurobehavior and gut microbiota community structure in mice. Methods: In August 2019, 64 C57BL/6 mice were randomly divided into 4 groups: control group (0 ppm) , low lead exposure group (20 mg/l) , medium lead exposure group (100 mg/l) and high lead exposure group (500 mg/l) . During the experiment, they were free to eat and drink. The drinking water of the lead exposure group was mixed with lead acetate, and sodium acetate was added in the control group. After 10 weeks of exposure, the Morris water maze was used to test the learning and memory ability of each group of mice, and then they were sacrificed for sampling. ICP-MS was used to detect lead content in whole blood and brain tissue. ELISA was used to determine the level of IL-1β in mouse serum. 16S rRNA sequencing was used to detect the structural diversity of the intestinal flora in feces, and then the correlation between the flora and behavior indicators was analyzed. Results: In the Morris water maze experiment, compared with the control group, there was no significant difference in the body weight and swimming speed of the mice in the lead exposure groups. The escape latency of the mice in the 100 mg/l and 500 mg/l dose groups was prolonged, and the number of platform crossings decreased (P<0.05) ; meanwhile, the staying time of the mice in the 500 mg/l Pb-treated group in the target quadrant was lower than that of the control group, and the difference was statistically significant (P<0.05) . Compared with the control group, the blood lead content of the mice in each lead exposure group was significantly increased, and the brain lead content of mice in the 500 mg/l dose group was significantly elevated (P<0.05) . The serum IL-1β levels of mice in each lead exposure group were higher than those of the control group (P<0.05) . At the phylum level, the relative abundance of the Proteobacteria phylum in all of Pb-treated groups was significantly increased (P<0.05) ; at the genus level, Allobaculum, Desulfovibrio, Lachnospiraceae_NK4A136_group, Turicibacter and Ureaplasma were significantly increased (P<0.05) . Among them. The relative abundance of Desuffaoibrio, Turici bacter, and Ureaplasma was negatively correlated with the residence time of mice in the quadrant of the platform (r=-0.32, -0.29, -0.44, P<0.05) . Conclusion: Lead exposure induced learning and memory impairments in mice, which may be related to the disturbance of the gut microbiota.

Heavy Metals Content in Whole Blood (Literature Review and Case Study)

Heavy metals content in whole blood was analyzed for Kyiv city residents, who were tested in 2019-2020 years. According to obtained results, the metals were divided into three group. 1) with elevated concentration in statistical sample and elevated threshold (mercury and arsenic); 2) with elevated concentrations for several patients while average concentration for sample did not exceed threshold (lead); 3) with single cases of increased concentrations that are not a risk to the health of the population, but pose a threat to a particular patient (Bi, Cd, Cr, Mo, Cs, Va). Some metals never exceeded the maximum allowable concentrations Al, Ba, Be, Au, Co, Cu, Mn, Ni, Pd, Pt, Sr, Ti, U, Zr). We recommend regular check-up for the concentrations of mercury and arsenic in blood, because these metals are the factors of population risk. When the metals concentrations in blood are elevated, the diagnosis should be established taking into account clinical history of the patient.

REGULATION OF THE MACROELEMENT METABOLISM IN COWS DEPENDING ON THE TYPE OF HIGHER NERVOUS ACTIVITY

REGULATION OF THE MACROELEMENT METABOLISM IN COWS DEPENDING ON THE TYPE OF HIGHER NERVOUS ACTIVITY

Monitoring of indicators of mineral metabolism in new zealand white rabbits

The quality of livestock products depends on the mineral status of the animals. The study of mineral elements makes it possible to investigate changes in the environment and to prevent negative impacts both on the animal organism, the wellbeing of which is completely dependent on humans, and on the human organism, as the final consumer of food of animal origin. A promising direction in veterinary medicine is the development of new methods of non-invasive diagnostics for assessing the state of health and determining metabolic disorders of essential elements in the body of rabbits, because these methods will be more humane and will facilitate the work of a veterinary medicine doctor. The aim of the study was to determine the characteristics of mineral metabolism in rabbits of the New Zealand white breed by age and sex and the informativeness of the assessment of metabolism for the use of various biological materials. The studies were carried out under production conditions on 24 clinically healthy rabbits of the New Zealand White breed, of all ages and sex. For research, blood, urine, and hair were taken from the animals. The content of trace elements in biological materials was determined by the method of atomic emission spectrometry with inductively coupled plasma. A significant correlation was revealed between the content in the blood plasma of rabbits of Calcium with hair (-0.42), Manganese with hair (-0.32), Iron with hair (0.63), Zinc with hair (0.40), Copper with hair (0.39) and with urine (0.44); between the content in whole blood of rabbits of Calcium with hair (0.47) and with urine (- 0.33), Manganese with hair (-0.32); between the content of Cobalt with hair in the urine (-0.35). The dynamics of the content of Calcium, Manganese, Iron, Zinc, Cobalt, and Copper in the biological materials of rabbits depending on their age and sex was found out, which is indicated by a significant increase in the blood plasma of Calcium, Manganese, Iron, Zinc, Cobalt, and Copper in females from 70 to 240 days and a significant increase in Manganese and Zinc and a decrease in Copper in males from 70 to 90 days. The informational content of the non-invasive method for diagnosing microelementoses in rabbits has been experimentally and theoretically substantiated, and the informational content of the microelement composition of hair for biogeocoenotic diagnostics has been proved, which is appropriate for medical examination and mass research of rabbits. In the future, we plan to investigate other elements in different biological materials of rabbits, followed by an assessment of the information content of noninvasive methods for diagnosing disorders of mineral metabolism.

The effect of photobiomodulation therapy of some indices of rats’ blood cells functional state under experimental diabetes mellitus

Background. Diabetes mellitus is a chronic endocrine-metabolic disease caused by an absolute or relative insulin deficiency. During diabetes, there are perfect conditions for the development of oxidative stress: the content of substrates for oxidation increases, the content of natural antioxidants decreases and the activity of antioxidant systems is suppressed. It is known that photobiomodulation therapy produce antioxidant and antihyperglycemic effects. Here we investigated its influence on blood system functioning. Materials and Methods. The study was performed on male Wistar rats. Experimental diabetes mellitus was induced by the intraperitoneal injection of streptozotocin. Leukocyte formula was calculated using blood smears stained by Romanowsky–Giemsa. Catalase activity was determined spectrophotometrically. Affinity of hemoglobin to oxygen was evaluated by spectrophotometric method in Ivanov’s modification by drawing oxygenation curves. The protoporphyrin content in whole blood was measured by analyzing its fluorescence spectra. The content of NO2-, total and inducible NO synthase activity was determined spectrophotometrically. Results. Under the action of photobiomodulation therapy on healthy animals, there was a shift of oxygenation curves to the left and a decrease of P50, whereas under irradiation of rats with diabetes, there was a shift of oxygenation curves to the right and increase in P50 compared to indices in nonirradiated animals. During diabetes, there was a decrease in protoporphyrin content compared to control, but there was a tendency to increase under photobiomodulation. Photobiomodulation therapy of rats with diabetes increased catalase activity in erythrocyte hemolysates. We revealed significant changes in leukocyte formula under photobiomodulation. The total NO synthase activity in leukocytes of rats with diabetes was higher compared to healthy animals, but decreased under the action of photobiomodulation. We found an increase in inducible NO synthase activity in leukocytes of rats with diabetes and in leukocytes of irradiated healthy animals. An increase in NO2- content in leukocytes of rats with diabetes was detected. Under photobiomodulation, NO2- content was significantly lower in rats with diabetes. Conclusion. Photobiomodulation therapy produces a corrective action on blood system during diabetes, in particular, it improves oxygen release from hemoglobin and prevents hypoxia. Simultaneously with the increase in tissue oxygen saturation, a decrease in NO synthase activity and nitrite content along with an increase in catalase activity prevents the development of oxidative stress.

ROLE OF THE AUTONOMIC NERVOUS SYSTEM IN THE REGULATION OF PHOSPHORUS AND CALCIUM METABOLISM IN COWS

The leading role in the mobilization of the organism adaptive capabilities is played by neuro- humoral mechanisms, primarily the activity of the central nervous system. The full-value mineral nutrition of cattle depends on the provision of animals with macronutrients, which are included in the structure of many enzymes or are their activators, taking a leading role in redox reactions. The studies were carried out on cows of the Ukrainian black-and-white breed of 2-3rd lactation. By the results of the study of tone of the autonomic nervous system, 3 research groups were formed: І – normotonic , ІІ – vagotonic , ІІІ – sympathicotonic cows. Blood from cattle was obtained twice a year in summer and winter. Whole blood was stabilized with heparin, blood serum was obtained by sedimentation method, and blood cells – by centrifugation of heparinized blood, plasma collection and after triple washing of cells in cold isotonic solution, followed by centrifugation. The tone of the autonomic nervous system in cows in summer is inversely related to the content of phosphorus in whole blood (r = -0.73; P&lt;0.05) and blood cells (r = -0.87; P&lt; 0.001). However, in blood serum these relationships are unreliable both in summer and in winter ( r= – 0 . ,02–0 . 24). In winter, the tone of the autonomic nervous system in cows is inversely reliable associated only with the phosphorus content in whole blood (r=-0.81; P &lt;0.01). Unlike the tone of the autonomic nervous system, the season has a significant effect on the calcium content in whole blood (F = 8.94&gt; FU = 8 .41; P&lt;0.01), while the content of this element in the serum and blood cells of cows did not depend from the season. So, the tone of the autonomic nervous system and the season affect the content of calcium and phosphorus in the blood of cows. Keywords: higher nervous activity, types of higher nervous activity, nervous processes , autonomic nervous activity,

CORTICAL MECHANISMS OF REGULATION OF LITHIUM CONTENT IN BLOOD OF COWS DEPENDING ON THE SEASONS

The behavior of farm animals, especially cattle, is an important economic and useful feature, which significantly facilitates the operating conditions and production of livestock products. The parameters of the nervous system in cattle determine its behavior in various situations, including keeping in the pen, herding, and milking. The optimal content of minerals in the body leads to the normal course of metabolic processes and high productivity. Based on the results of studying the conditioned reflex activity, four experimental groups were formed. Whole blood, serum, and blood cells were used for laboratory analysis. Based on the results of the analysis of variance of the lithium content in different blood fractions of cows, no reliable interaction between the typological features of the nervous system and the season has been established. The mobility of nervous processes, regardless of the season, is reliably associated with the lithium content in whole blood, blood cells, and blood serum (r = 0.15–0.22). Along with this, the parameter of the lithium transmembrane potential (Li cell /Li serum ) is reliably associated mainly with the strength and balance of nervous processes in cows in the warm season – r = 0.62–0.76 (P &lt; 0.05–0.01). A significant dependence on the type of higher nervous activity in cows and the lithium content in blood serum (F = 13.2 &gt; FU = 3.01; P &lt; 0.001), whole blood (F = 14.8 &gt; FU = 3.01; P &lt; 0.001), and blood cells (F = 23.1 &gt; FU = 3.01; P &lt; 0.001) is established. In cows with a weak type of higher nervous activity, the lithium content in blood serum and its cells was lower by 12.1% (P &lt; 0.05) and 18.0% (P &lt; 0.05), respectively, compared with the parameters of cows with a strong balanced mobile type. Keywords: higher nervous activity, micronutrients, types of higher nervous activity, nervous processes

Establishment of a Patient-Derived Xenograft Model of Colorectal Cancer in CIEA NOG Mice and Exploring Smartfish Liquid Diet as a Source of Omega-3 Fatty Acids.

Cancer patient-derived xenografts (PDXs) better preserve tumor characteristics and microenvironment than traditional cancer cell line derived xenografts and are becoming a valuable model in translational cancer research and personalized medicine. We have established a PDX model for colorectal cancer (CRC) in CIEA NOG mice with a 50% engraftment rate. Tumor fragments from patients with CRC (n = 5) were engrafted in four mice per tumor (n = 20). Mice with established PDXs received a liquid diet enriched with fish oil or placebo, and fatty acid profiling was performed to measure fatty acid content in whole blood. Moreover, a biobank consisting of tissue and blood samples from patients was established. Histology, immunohistochemistry and in situ hybridization procedures were used for staining of tumor and xenograft tissue slides. Results demonstrate that key histological characteristics of the patients’ tumors were retained in the established PDXs, and the liquid diets were consumed as intended by the mice. Some of the older mice developed lymphomas that originated from human Ki67+, CD45+, and EBV+ lymphoid cells. We present a detailed description of the process and methodology, as well as possible issues that may arise, to refine the method and improve PDX engraftment rate for future studies. The established PDX model for CRC can be used for exploring different cancer treatment regimes, and liquid diets enriched with fish oil may be successfully delivered to the mice through the drinking flasks.

Biochemical Dymorphism of Potassium Content and its Adaptive Significance in Domestic Reindeer

The connection between the types of potassium concentration and the ecogenesis of the studied groups of animals, with the degree of extreme conditions of breeding, with types of potassium in extreme conditions of breeding is known. In this regard, studies on autochthonous species of ungulates in the Subarctic are of great interest, the most numerous of which is the reindeer (Rangifer tarandus). Comparative analysis of the adaptive responses of NK and LK types at different levels of organization confirmed the presence of the adaptive advantage of animals characterized by a hereditarily high concentration of potassium in the blood in ecological-geographical zones with extreme environmental conditions. As a result of a study on domesticated reindeer, a clearly expressed dimorphism in the level of potassium in the blood was revealed, due to a genetically controlled high NK and low LK concentration. The average level of potassium in whole blood in the studied population of reindeer is 26.94 ± 0.56 meq/l with a variability of 24.61%. It was revealed that the average level of potassium content in whole blood by sex and age was distributed as follows: whales 22.97 - 25.80 meq/l at P&gt; 0.05, castrate bulls had indicators significantly higher than 28.37 - 29.58 meq / l, at P&gt; 0.05, the young showed intermediate indicators - 27.37 - 28.13 at P&gt; 0.05. The intragroup variability in potassium content in whole blood ranged from 15.5 to 31.05%. It was found that in domestic reindeer the ratio of animals of the high-potassium (HK) type is 88.02% and of the lowpotassium (LK) type - 11.98%, while the gene frequencies for highpotassium animals are 0.9381.

Взаємозв’язок основних характеристик нервових процесів із вмістом натрію у крові корів залежно від пори року

The type of higher nervous activity determines individual differences and the body's ability to adapt to changing environmental conditions. The most perfect adaptation is provided by a combination of high strength, mobility and balance of nervous processes. It was found that the strength of nervous processes both in summer and in winter was inversely related only to the sodium content in blood cells (r = -0.57–0.71; p&lt;0.01). The balance of excitation and inhibition processes in the cerebral cortex in summer was inversely correlated with the sodium content in whole blood (r = -0.56; p&lt;0.05) and blood cells (r = -0.64; p &lt;0.01), in winter – in blood serum of cows (r = 0.62; p &lt;0.01). It is proved that in summer the effect of the balance on the sodium content in whole blood was ղ²х = 0.29 (p &lt;0.05), in blood cells – ղ²х = 0.63 (p&lt;0.001), and in winter in blood cells – ղ²х = 0.37 (p&lt;0.05). In summer, the mobility of nervous processes to a greater extent limited the sodium content in blood of cows than in winter. Thus, the effect of this indicator of cortical regulation on the sodium content in whole blood and blood cells in summer reached ղ²х = 0.25–0.35 (p&lt;0.05), and in winter in blood cells – ղ²х = 0.24 (p&lt;0.05).

Взаємозв’язок тонусу автономної нервової системи у корів з вмістом купруму в крові залежно від пори року

The tone of the autonomic nervous system is considered on the one hand as one of the manifestations of homeostasis, and on the other hand, as one of the mechanisms of its regulation. The predominance of the tonic effect of the parasympathetic and sympathetic departments of the autonomic nervous system is defined as vagotonia and sympathicotonia. The tone of the autonomic nervous system in cows was determined using a trigeminal vagal test. According to the results, the animal was classified as normotonic, sympathicotonic or vagotonic. The tone of the autonomic nervous system in cows in summer inversely correlated with copper content in whole blood (r=-0.62; p&lt;0.05) and blood cells (r=-0.58; p&lt;0.05). In winter, the correlation of the tone of the autonomic nervous system in cows with its content in whole blood, serum and blood cells did not reach significant values (r=-0.21–0.36). Increased tone of the parasympathetic department of the autonomic nervous system, regardless of the season, had almost no effect on copper content in serum, whole blood and its cells (ղ²x=0.01–0.23). At the same time, sympathicotonia in summer has a significant impact only on the copper content in blood cells – ղ²x=0.58 (p&lt;0.05), while in winter this effect increased significantly (ղ²x=0.91; p&lt;0.001). The multifactor analysis of variance revealed the dependence of copper content on the tone of the autonomic nervous system and the season. However, the vegetative status of cows had a significant impact on its content only in blood cells (F=8.59&gt;FU=4.41; p&lt;0.01), and the season limited copper content only in whole blood (F=12.12&gt;FU=3.55; p&lt;0.01).

Health Outcomes at School Age among Children Who Are HIV-Exposed but Uninfected with Detected Mitochondrial DNA Depletion at One Year

Infant antiretroviral (ARV) prophylaxis given to children who are human immunodeficiency virus (HIV)-exposed but uninfected (CHEU) to prevent HIV transmission through breastfeeding previously proved its efficacy in the fight against the pediatric epidemic. However, few studies have investigated the short- and long-term safety of prophylactic regimens. We previously reported a decrease of mitochondrial DNA (mtDNA) content among CHEU who received one year of lamivudine (3TC) or lopinavir-boosted ritonavir (LPV/r) as infant prophylaxis. We aimed to describe mtDNA content at six years of age among these CHEU, including those for whom we identified mtDNA depletion at week 50 (decrease superior or equal to 50% from baseline), and to compare the two prophylactic drugs. We also addressed the association between mtDNA depletion at week 50 with growth, clinical, and neuropsychological outcomes at year 6. Quantitative PCR was used to measure mtDNA content in whole blood of CHEU seven days after birth, at week 50, and at year 6. Among CHEU with identified mtDNA depletion at week 50 (n = 17), only one had a persistent mtDNA content decrease at year 6. No difference between prophylactic drugs was observed. mtDNA depletion was not associated with growth, clinical, or neuropsychological outcomes at year 6. This study brought reassuring data concerning the safety of infant 3TC or LPV/r prophylaxis.

Quantification of circadian rhythm in mitochondrial DNA copy number in whole blood, and identification of factors that influence it

Mitochondrial DNA (mtDNA), the genetic material in mitochondria, encodes key genes related to the respiratory chain and ATP production. To accurate quantification mtDNA content in whole blood is important for various disease states. Absolute quantitative Real-time PCR and platelet contamination erase method were used for mtDNA copy number analysis in whole blood. In the quantitative study of mtDNA content, it was found that whole blood mtDNA copy number showed a fluctuating rhythm during a 24-h period due to dynamic changes in white blood cells combined with platelets. However, when isolated white blood cells were used, or absolute whole blood mtDNA was calculated, the circadian rhythm pattern of mtDNA disappeared. In this study, a feasible method that can accurately quantify mitochondrial DNA in small blood samples was established, and it was found that two factors which greatly influenced mtDNA copy number were sampling time and platelets in blood.

Techniques of quantitative evaluation of verapamil content in whole blood

Verapamil is a drug that can be toxic in pharmacological therapy and in case of misuse. Simple and informative methods of Verapamil quantification for forensic chemistry and hospital toxicology are needed.&#x0D; Aim. The objective of the study was to compare analytical potential of different methods for Verapamil quantification used in forensic chemistry and hospital toxicology.&#x0D; Materials and methods. The subject study was whole blood samples containing Verapamil. Verapamil in the blood samples was identified by gas chromatography-mass-spectrometry. Verapamil was quantified by thin-layer chromatography with videodensitometry and UV-spectrophotometry.&#x0D; Results. To quantify Verapamil content, the plates were scanned, the chromatogram images were processed and calibration models were given by means of computer program. The calibration model is described by polynomial (square) regression. In UV-spectrophotometry absorbance of samples at the wavelength of 277 nm was measured; blank blood extracts as a zero reference were used. The limits of quantifation for thin-layer chromatography with videodensitometry and UV-spectrophotometry were 300,0 and 6000,0 ng/ml respectively. The accuracy and precision for thin-layer chromatography with videodensitometry failed to exceed 20.2 и 24.3% respectively; for UV-spectrophotometry they were 27.5 и 7.4%.&#x0D; Conclusion. The ranges of quantifation make it possible to use thin-layer chromatography with videodensitometry for forensic chemistry and hospital toxicology. UV-spectrophotometry can be used to quantify Verapamil content in blood samples at the lethal concentration range.