Technology of Quick-Frozen Flour Fish Culinary Products

In this article, the results of the development of the functional fish culinary product “Thorny Skate and Cod Pie” are presented. A traditional recipe was used for making the yeast dough for the pie. The pie filling recipe was designed using Fuzzy Logic in the Matlab software package.Optimized parametersfor the selected sensory evaluation of the pie were calculated. On the basis of a priori information, key components of the filling (including the fraction of the fish components and skate meat) were chosen as the factors of interest. According to the simulation results, the optimal values werea 50/50 percentage for the first and the second factor respectively, and this providedthe maximum organoleptic assessment (five points on a five-point scale). The simulation results were compared with the results of the organoleptic evaluation of the pie made according to the optimized recipe, and their sufficient convergence was shown. The indicators of mass fraction of amine nitrogen and nitrogen of volatile bases was studied, as well as the microbiological safety indicators of flour fish culinary products, in accordance with the requirements of the Technical Regulations of the Eurasian Economic Union 040/2016 ”On the safety of fish products”. The results showed a high efficiency of the shock freezing of the semi-finished product, brought to semi-readiness, for long-term storage (120 days at a temperature no higher than minus 18 ∘C), without reducing the quality or safety of the pie. The product had a cholesterol content from 220 to 260 mg%, which allowed it to be classified as functional. The nutritional values of the product (mass fraction of protein, fat, carbohydrates, and amino acid composition) are presented. Keywords: thornyskate, functional product, pie with thornyskate and cod, shock freezing

Compression enhances invasive phenotype and matrix degradation of breast Cancer cells via Piezo1 activation

Background Uncontrolled growth in solid breast cancer generates mechanical compression that may drive the cancer cells into a more invasive phenotype, but little is known about how such compression affects the key events and corresponding regulatory mechanisms associated with invasion of breast cancer cells including cellular behaviors and matrix degradation. Results Here we show that compression enhanced invasion and matrix degradation of breast cancer cells. We also identified Piezo1 as the putative mechanosensitive cellular component that transmitted compression to not only enhance the invasive phenotype, but also induce calcium influx and downstream Src signaling. Furthermore, we demonstrated that Piezo1 was mainly localized in caveolae, and both Piezo1 expression and compression-enhanced invasive phenotype of the breast cancer cells were reduced when caveolar integrity was compromised by either knocking down caveolin1 expression or depleting cholesterol content. Conclusions Taken together, our data indicate that mechanical compression activates Piezo1 channels to mediate enhanced breast cancer cell invasion, which involves both cellular events and matrix degradation. This may be a critical mechanotransduction pathway during breast cancer metastasis, and thus potentially a novel therapeutic target for the disease.

Protein-synthesizing, bile-forming, urea-forming and carbohydrate functions in cows with fatty degeneration of the liver

In dairy farms of Ukraine, where highly productive dairy cows are kept, liver lesions are often diagnosed in the postpartum period. Postmortem studies of the liver of cows that were forcibly slaughtered showed that in mostly animals were diagnosed with fatty degeneration of the liver. The main causes of fatty hepatosis were violations of the structure of rations, imbalance of feeding on essential nutrients and biologically active substances, low content of easily digestible carbohydrates and high protein content. The study was performed on cows aged 4–5 years with productivity for the previous lactation of 5.600–7.500 L of milk, in a winter-stall period of keeping, 2–3 weeks after calving. According to clinical and biochemical blood tests, two groups of cows were formed – 50 clinically healthy and 50 cows with fatty liver disease. In cows diagnosed with fatty liver degeneration, the disease was manifested by decreased productivity and fatness, loss of appetite, oppression, hypotony of the rumen, reticulum and omasum. In some cows, there was pain at the liver area, increasing boundaries of hepatic dullness, jaundice of the visible mucous membranes and sclera. The blood serum of all cows with fatty liver disease established a decrease in albumin content, indicating impaired protein synthesis function of the liver. In some cows, the content of total protein in the serum increased due to globulin fractions, mainly gamma globulins. The ratio between the content of albumins and globulins decreased, which indicates the development in the blood of sick animals dysproteinemia. The development of fatty infiltration of the liver caused an increase in the concentration of bile acids in the serum of all sick cows. This is due to reduced conjugation and excretion of cholates by affected hepatocytes from the bile capillaries. The formation, absorption, conjugation, and excretion of bilirubin in the bile is disturbed, which causes the accumulation of total and conjugated bilirubin in the serum of sick animals. The cholesterol content in the blood of cows decreased, caused a violation of the esterification of its esters by hepatocytes. The established changes in the content of bile acids, total and conjugated bilirubin, and cholesterol in the blood of sick cows indicate a violation of bile secretion, bile production, and cholestasis development. In some cows with fatty liver degeneration, urea formation function and carbohydrate function are impaired, leading to a decrease in blood urea content and glucose.

Cyclophilin A Impairs Efferocytosis and Accelerates Atherosclerosis by Overexpressing CD 47 and Down-Regulating Calreticulin

Impairment of efferocytosis in apoptotic macrophages is a known determinant of the severity of atherosclerosis and the vulnerability of plaques to rupture. The precise mechanisms involved in impaired efferocytosis are unclear. Given the well-recognized role of the inflammatory cytokine cyclophilin A (Cyp A) in modulating several atherogenic mechanisms in high-glucose primed monocytes, we investigated the role of Cyp A in macrophage efferocytosis. The efficiency of efferocytosis in RAW 264.7 macrophages grown in vitro and primed with cyclophilin A was assessed using flow cytometry and confocal assays. Cholesterol content in cells was measured using cell-based cholesterol efflux assay. Proteomic analysis and bioinformatics tools were employed to decipher the link between cyclophilin A and the known ligand receptors involved in efferocytosis. Cyclophilin A was found to impair efferocytosis in apoptotic macrophages by reducing ABCA1-mediated cholesterol efflux in foam cells derived from macrophages. Cyclophilin A-primed macrophages showed an increase in expression of the don’t-eat-me signal CD 47 and a decrease in the expression of the eat-me signal, calreticulin. Phagocytosis was restored upon silencing of cyclophilin A. New Zealand white rabbits were fed a high-fat diet, and lesions in their aortae were analyzed histologically for evidence of atherosclerosis and the expression of Cyp A, CD 47 and calreticulin, the ligand receptor involved in efferocytosis. Gene and protein expressions in aortae and macrophages were analyzed by real-time PCR and Western blotting. Cyclophilin A, via its effects on the expression of CD 47 and calreticulin, impairs efferocytosis in apoptotic macrophages. Together with its impact on cholesterol efflux from macrophages, these effects can amplify other mechanisms of Cyp A in accelerating the progression of atherosclerosis.

7-Ketocholesterol Induces Lipid Metabolic Reprogramming and Enhances Cholesterol Ester Accumulation in Cardiac Cells

7-Ketocholesterol (7KCh) is a major oxidized cholesterol product abundant in lipoprotein deposits and atherosclerotic plaques. Our previous study has shown that 7KCh accumulates in erythrocytes of heart failure patients, and further investigation centered on how 7KCh may affect metabolism in cardiomyocytes. We applied metabolomics to study the metabolic changes in cardiac cell line HL-1 after treatment with 7KCh. Mevalonic acid (MVA) pathway-derived metabolites, such as farnesyl-pyrophosphate and geranylgeranyl-pyrophosphate, phospholipids, and triacylglycerols levels significantly declined, while the levels of lysophospholipids, such as lysophosphatidylcholines (lysoPCs) and lysophosphatidylethanolamines (lysoPEs), considerably increased in 7KCh-treated cells. Furthermore, the cholesterol content showed no significant change, but the production of cholesteryl esters was enhanced in the treated cells. To explore the possible mechanisms, we applied mRNA-sequencing (mRNA-seq) to study genes differentially expressed in 7KCh-treated cells. The transcriptomic analysis revealed that genes involved in lipid metabolic processes, including MVA biosynthesis and cholesterol transport and esterification, were differentially expressed in treated cells. Integrated analysis of both metabolomic and transcriptomic data suggests that 7KCh induces cholesteryl ester accumulation and reprogramming of lipid metabolism through altered transcription of such genes as sterol O-acyltransferase- and phospholipase A2-encoding genes. The 7KCh-induced reprogramming of lipid metabolism in cardiac cells may be implicated in the pathogenesis of cardiovascular diseases.

Targeting cytochrome P450 46A1 and brain cholesterol 24-hydroxylation to treat neurodegenerative diseases

The brain cholesterol content is determined by the balance between the pathways of in situ biosynthesis and cholesterol elimination via 24-hydroxylation catalyzed by cytochrome P450 46A1 (CYP46A1). Both pathways are tightly coupled and determine the rate of brain cholesterol turnover. Evidence is accumulating that modulation of CYP46A1 activity by gene therapy or pharmacologic means could be beneficial in the case of neurodegenerative and other brain diseases and affect brain processes other than cholesterol biosynthesis and elimination. This minireview summarizes these other processes, most common of which include abnormal protein accumulation, memory, and cognition, motor behavior, gene transcription, protein phosphorylation as well as autophagy and lysosomal processing. The unifying mechanisms, by which these processes could be affected by CYP46A targeting are also discussed.

INFLUENCE OF DIFFERENT DOSES OF TISSUE BIOSTIMULANT ON BIOCHEMICAL BLOOD INDICES OF YOUNG CATTLE

Изучено влияние введения разных доз тканевого биостимулятора на биохимические показатели сыворотки крови ремонтного молодняка крупного рогатого скота. Опыт проводился в Алтайском крае на 4 группах ремонтных телочек Приобского типа черно-пестрой породы по 10 голов в каждой. При подборе животных учитывались возраст (1 мес) и живая масса (51,3±1,48 кг). Продолжительность опыта составляла 14 дней. Животным контрольной группы подкожно однократно вводили физиологический раствор в дозе 3,0 мл на 1 голову, I опытной — тканевый биостимулятор в дозе 2,0 мл, II — 3,0 мл, III — 4,0 мл на 1 голову. Опытную партию тканевого биостимулятора изготовили из субпродуктов и боенских отходов пантовых оленей по запатентованной технологии. Материалом для приготовления препарата служили мезентеральные лимфоузлы и средостения, селезенка, печень, матки с плодами (2—3 мес), плацента, отобранные в асептических условиях во время убоя здоровых животных. Введение тканевого биостимулятора телочкам в разных дозах способствовало повышению некоторых исследуемых биохимических показателей сыворотки крови. Наиболее оптимальной дозой применения тканевого биостимулятора следует считать 3,0 мл/гол, что способствует повышению содержания общего белка в сыворотке крови на 1,4% (P≤0,05), глюкозы — на 22,6% (P≤0,05) и снижению содержания холестерина на 12,3% (P≤0,05). The effect of the administration of different doses of the tissue biostimulant on the biochemical blood serum indices of replacement young cattle was studied. The experiment was carried out in the Altai Region in 4 groups of 10 Black-Pied replacement heifers of the Priobskiy type. When selecting the animals, their age (1 month) and live weight (51.3±1.48 kg) was taken into account. The experiment lasted 14 days. Saline solution was injected under the skin to the animals of the control group at a dose of 3.0 mL per 1 head; the tissue bio-stimulant was administered in the following doses: in the 1st trial group — 2.0 mL per head; the 2nd trial group 3.0 mL per head; the 3rd trial group — 4.0 mL per 1 head. The trial batch of the tissue bio-stimulant was made from velvet antler deer by-products and slaughterhouse offal by using the patent-protected technology. The tissue biostimulant was made from mesenteric lymph nodes and mediastinums, spleen, liver, uteri with 2—3 month old fetuses, and placentae collected under aseptic conditions from healthy animals at slaughter. The administration of the tissue bio-stimulant to heifers in different doses increased some of the studied biochemical blood serum indices. The tissue biostimulant dose of 3.0 mL per head should be considered the most optimal one; it increased the total protein content in blood serum by 1.4% (P≤0.05), glucose — by 22.6% (P≤0.05), and decreased cholesterol content by 12.3% (P≤0.05).

The Effect of L-Glutamine on Basal Albumen and Yolk Indices and Albumen Amino Acids Composition

The current study tested the hypothesis that 1.0% dietary inclusion of L-glutamine (Gln), an non-essential amino acid that influences protein synthesis, can improve internal egg quality, including amino acids profile. Thirty-week-old Bovans Brown laying hens in their middle laying period were assigned to one of the two experimental groups (12 replicate cages, 2 hens/cage) with Gln in the form of alpha-ketoglutarate (10 g/kg) or without Gln inclusion. The experimental period lasted for 30 wks, from the 31st to the 60th week of age of hens, when eggs were collected and selected egg quality indices were determined. Gln supplementation had no effect on albumen and egg yolk share, albumen and yolk basal indices and composition, including yolk cholesterol content. However, Gln decreased the lipid content of the egg albumen (p < 0.001), and influenced albumen amino acid profile, increasing content of asparagine (p < 0.05), phenylalanine (p < 0.05), proline (p < 0.001), tryptophan (p < 0.01), and tyrosine (p < 0.05). In conclusion, the study shows a potential role of Gln supplementation for enhancing nutritional values of eggs by lower lipid content and higher amino acid profile.

Wnt5a Promotes Lysosomal Cholesterol Egress and Protects Against Atherosclerosis

Background: Impairment of cellular cholesterol trafficking is at the heart of atherosclerotic lesions formation. This involves egress of cholesterol from the lysosomes and two lysosomal proteins, the Niemann-Pick C1 (NPC1) and NPC2 that promotes cholesterol trafficking. However, movement of cholesterol out the lysosome and how disrupted cholesterol trafficking leads to atherosclerosis is unclear. As the Wnt ligand, Wnt5a inhibits the intracellular accumulation of cholesterol in multiple cell types, we tested whether Wnt5a interacts with the lysosomal cholesterol export machinery and studied its role in atherosclerotic lesions formation. Methods: We generated mice deleted for the Wnt5a gene in vascular smooth muscle cells (VSMCs). To establish whether Wnt5a also protects against cholesterol accumulation in human VSMCs, we used a CRISPR/Cas9 guided nuclease approach to generate human VSMCs knockout for Wnt5a. Results: We show that Wnt5a is a crucial component of the lysosomal cholesterol export machinery. By increasing lysosomal acid lipase expression, decreasing metabolic signaling by the mTORC1 kinase, and through binding to NPC1 and NPC2, Wnt5a senses changes in dietary cholesterol supply and promotes lysosomal cholesterol egress to the endoplasmic reticulum (ER). Consequently, loss of Wnt5a decoupled mTORC1 from variations in lysosomal sterol levels, disrupted lysosomal function, decreased cholesterol content in the ER, and promoted atherosclerosis. Conclusions: These results reveal an unexpected function of the Wnt5a pathway as essential for maintaining cholesterol homeostasis in vivo.