Blood Samples Of Animals Research Articles

Evaluation of fenugreek (Trigonella foenum-graecum L.) powder supplementation on metabolic syndrome, oxidative stress and inflammation in high fat diet fed rats

As an anti-diabetic medicinal plant, fenugreek seed (FG) (Trigonella foenum-graecum L.) has long been utilized and has many therapeutic uses. The current investigation sought to ascertain the effects of FG powder supplementation on insulin resistance, dyslipidemia, and oxidative stress in rats given a high-fat (HF) diet. Measurements of biochemical and antioxidant indicators were made in animal blood and tissue samples, including malondialdehyde (MDA), nitric oxide (NO), advanced oxidation protein product (AOPP), reduced glutathione (GSH), superoxide dismutase (SOD), triglycerides (TG), and catalase (CAT). Furthermore, the tissues of the kidney, heart, and liver were stained histologically. HF diet feeding in rats declined in antioxidant enzyme activities, along with a rise in MDA and other indicators of oxidative stress, which were mitigated by the supplementation of FG. Additionally, FG supplementation enhanced the expression of genes corresponding to antioxidants in the liver of rats given HF diet. FG supplements in HF diet-fed rats displayed altered gene expression in the livers that metabolized fat. Histological staining of the liver demonstrated that FG supplementation reduced necrosis and the accumulation of fat droplets in the liver of rats given HF diets. In conclusion, this finding showed that FG supplementation in HF diet fed rats reduced the plasma lipids, decreased oxidative stress and raised antioxidant enzyme activities. FG may therefore aid in reducing the chronic complications linked to the HF diet in experimental rats.

Seroprevalence and contributing factors of transboundary animal diseases in sheep and goats: a study in Peninsular Malaysia.

Diseases caused by small ruminant lentiviruses, Mycobacterium avium ssp. paratuberculosis (MAP), Schmallenberg virus, and peste des petits ruminants virus (PPR) is globally recognised as serious threats to the ruminant industry due to their potential to spread rapidly across boundaries. Despite their global distribution and negative impacts on ruminant production, there is a gap in knowledge of the current trends in their epidemiology among sheep and goat populations in Peninsular Malaysia. This study was therefore designed to fill the gap of knowledge concerning the seroprevalence and contributing factors of CAEV, paratuberculosis, SBV, and PPRV among small ruminants from selected flocks in Selangor, Negeri Sembilan, and Pahang states in Peninsular Malaysia. A cross-sectional study design was used to collect animal data and blood samples for serological assays simultaneously. The ID Screen (ID.VET, France) indirect ELISA screening tests were used to detect serum antibodies directed against CAEV/MVV (VISNAS Ver 0922), paratuberculosis (PARAS Ver 0516), SBV (SBVC Ver 1114) and PPRV (PPRC Ver 0821). There was 45.4% (95% CI = 40.74-50.74), 6.8% (95% CI = 4.66-9.69), 27.8% (95% CI = 23.35-32.77), and 2.6% (95% CI = 1.11-0.51) true seroprevalence for CAEV, paratuberculosis, SBV, and PPR, respectively. Geographical location and species were the risk factors for CAEV and paratuberculosis, while the management system and age of small ruminants were the risk factors for SBV. The present study is the first to document a large-scale seroprevalence of MAP and PPR infection among sheep and goat flocks in Peninsular Malaysia. The presence of PPRV and MAP antibodies among small ruminant flocks is signalling current or previous exposure to the pathogens or cross reactions with similar antigens. This finding further suggests the potential for future outbreaks of these devastating diseases among sheep and goats in Malaysia. The high seroprevalence of CAEV and SBV among small ruminants indicates high levels of exposure to the viruses in the environment, which is a potential threat to production.

The epizootic situation of bovine leukemia in the Republic of Dagestan in 2023

Bovine enzootic leukemia (EBLV) remains an acute problem for animal husbandry in many regions of Russia. Based on this, the goal was set: to study the epizootic situation of bovine leukemia in the Republic of Dagestan. The material for the study was taken from the official source of the Veterinary Committee of the Republic, as well as supplemented with our own serological blood tests for bovine leukemia virus (bovine leukemia). Laboratory tests for the detection of antibodies to BLV in RID were carried out according to the "Guidelines for the diagnosis of bovine leukemia" (2000). Research results: the spread of BLV has been established in some municipal districts and urban districts throughout the republic. In 2023, from all veterinary laboratories (stations), serological studies were conducted in the amount of 1101,689 animal blood samples for BLV. Of the animals studied, antibodies to the BLV antigen in RID were obtained in 2,778 (0.25%) samples. A high percentage of animal HCV infection was detected in the following districts and cities: Kizlyarsky – 2% (388), Tlaratinsky – 1.3% (341), Botlikhsky – 1.2% (450), Babayurt – 1.1% (188), Gunibsky – 0.8% (228) and in Makhachkala – 0.9% (180). In 16 districts and in Khasavyurt, the percentage of infection of animals with BLV was at the level of 0.3% or lower. We obtained 440 animal blood samples from sample farms, which were examined in the RID. 37 (8.4%) animal blood samples gave a positive result for BLV. Thus, the general epizootic situation in EBLV remains relevant, but the prevalence of BLV among animals is decreasing in the republic.

Immunobiological status of the organism of young pigs during their triphasic period cultivation and actions of corrective factors

The problem of the impact and consequences of technological stress is accompanied by several reactions that do not always have a positive effect on the animal's body. The work aimed to investigate the dynamics of indicators of the immunobiological status of piglets during their three-phase rearing and the use of immunostimulating agents Globigen® Pig Doser and Globigen® Jump Start. Parallel studies were conducted based on the private farm “Meat Resources” LLC of the Lviv region of the Starosambir district, which practices three-phase pig breeding. The experiment involved 36 analog piglets and hybrids of Landrace and Great White breeds, which were formed into 3 groups of 12 animals each. Piglets of the control group were fed the standard ration provided by the technology. During the first three days of life, newborn piglets of the first experimental group were given 2 ml/gal of liquid Globigen®Pig Doser. The piglets of the second experimental group were given a powdered dry product Globigen®Jump Start, which was added to the main diet at 0.2 % per ton of feed. It was established that the use of means with an immunostimulating effect was accompanied by a decrease in the content of Ig M in the blood samples of experimental groups of piglets. Thus, in the blood serum of 30-day-old piglets under the influence of Globigen® Pig Doser with a high level of probability (Р < 0.02), a decrease in the content of this class of antibodies was found to the level of 1.59 mg/ml, and under the influence of Globigen® Jump Start the content in the blood serum of 60-day-old piglets, having reached the lowest value, was 2.08 mg/ml. The positive effect of immunostimulating agents, which was characterized by an increase in the amount of Ig G, was found in experimental groups of pigs at the age of 60 days: 17.10 mg/ml for the effects of Globigen® Pig Doser and 17.08 mg/ml for the impact of Globigen® Jump Start. The highest content of Ig A with high probability (P < 0.001) was found in blood samples of animals that were given Globigen® Jump Start – 3.45 mg/ml and 3.86 mg/ml under the influence of Globigen® Pig Doser. The practicality and effectiveness of the use of means with immunostimulating properties, the main active ingredient of which is the immunoglobulin of a chicken egg, is explained by the ability to influence the pathogenesis of diseases of the gastrointestinal tract, which arise as a result of weakening the resistance of the animal body under the influence of a large number of technological stresses. Research data prove that the specificity of the action of Ig Y has a positive effect on the preservation of the integrity of the villi of the intestinal mucosa, which is reflected by the strengthening of its protective and barrier function, which in general contributes to the increase in the percentage of animal survival: using Globigen®Pig Doser 80.0 % and 86.7 % using Globigen®Jump Start. Based on such results, it can be asserted that the use of Globigen® Pig Doser and Globigen® Jump Start for three-phase pig farming is scientifically justified and economically feasible.

Seromonitoring, Serotyping and Histopathology of Foot and Mouth Disease in Mithun (Bos Frontalis) in the Northeastern Hilly Terrain of India

Frequent FMD outbreaks have been one of the major challenges faced by the farmers and breeders of Mithun animals in Nagaland, owing to a lack of adequate knowledge and proper vaccination programs for the disease. Therefore, this study aimed to determine the extent of FMD serotype O virus infection, which is the most common serotype causing FMD specifically in the Mithun population in the northeastern state of Nagaland, India. In-vitro screening of serotype O virus antibodies in the blood serum of the Mithun population was carried out by ELISA kit. A total of 213 blood samples of Mithun animals were collected and screened where 67% of the samples tested were found to be positive and only 32% of the samples were found to be negative for the virus. All the samples collected from the neighbouring states of Mizoram and Arunachal Pradesh were found to be negative for the antibodies against the serotype O virus. Through m-RT PCR, O serotype was detected from all the clinically affected animals from the cells of the tongue epithelium, blood, and saliva of animals. During the histopathological studies, in the pericardium, severe interstitial myocarditis and liver showed focal areas of coagulative necrosis, focal necrosis, and pulmonary oedema were prominent and ballooning degeneration of epithelium in the tongue was evident in all cases. In conclusion, we were able to show the prevalence of FMDV serotype O antibodies in a very high percentage of the Mithun population that were tested along with pathological alterations, which should be addressed immediately with proper vaccination programs and by informing and educating the farmers and breeders about the disease before an outbreak occurs.

The discovery of human Plasmodium among domestic animals in West Sumba and Fakfak, Indonesia

Background In Indonesia, malaria incidence is at a high rate despite maximum preventive efforts. Therefore, this study aims to determine the possibility of a Plasmodium reservoir among domestic animals in malaria-endemic areas. Methods Animal blood was collected using EDTA tubes, then smeared and stained with Giemsa for Plasmodium microscopic identification. About 10 μl of blood was dropped on to a filter paper to capture Plasmodium DNA. Nested PCR was used for parasite molecular detection, while Plasmodium species were identified using the sequenced DNA. Results A total of 208 and 62 animal blood samples were collected from Gaura village, West Sumba and Fakfak village, West Papua, Indonesia respectively. In total, 32 samples from Gaura contained P. falciparum or P. vivax, while the Plasmodium percentage in buffalo, horse, goat, and dogs were 20.7%, 14.3%, 5.8%, 16.7%, respectively. P. knowlesi was not found in any of the samples, and no other species were detected in 18 pig blood samples. Conclusion Human Plasmodium existence among domestic animals in Indonesia partly explains the high prevalence and persistence of malaria in some endemic areas due to a reservoir host presence. Therefore, future studies need to ascertain the cause.

Investigation of the Presence of Bovine Papillomavirus in Blood Leukocytes of Cattle in West Mediterranean of Turkey

One hundred Holstein cattle, varying in age from one to ten years, who had warts on different parts of its bodies and had previously had Bovine papillomavirus (BPV) typing had blood samples taken from them from various managements in Burdur-Center and its surroundings. DNA was extracted from leukocytes using the DNeasy Blood & Tissue Kit (Qiagen, Germany). RT-PCR test was performed using consensus and type-specific primers for the detection of BPV types. As a result of the study, RT-PCR test and consensus primers were not detected in blood leukocyte samples. Through the use of type-specific primers in an RT-PCR assay, the viral genome components of BPV-10 (56), BPV-11 (1), BPV-10+BPV-11 (2), BPV-10+BPV-12, and BPV-10+BPV-14 (2) were detected in these samples. BPV-12 was detected for the first time in blood leucocytes. In blood samples taken from animals classified according to the anatomical regions where the warts are located the highest BPV positivity (22,2%) was detected in animals with warts on the teats. Also, the most BPV-10 type was determined in the blood samples of animals with wart lesions on the teat and lobes. When the distribution of BPV PCR positive samples according to age groups was made, BPV PCR positive was found mostly in the between 1 and 47 month age group (71,4%). It was determined that the prevalence of BPV was high in the between 1 and 47 age group, while the rates of 48-83 months and later decreased with increasing age. In this investigation, it was found that BPV type 10 had been prevalent in blood leukocyte samples from cattle with warts in various areas of the body and with variable macroscopic appearance.

Ameliorative effects of Atriplex crassifolia (C.A.Mey) on pain and inflammation through modulation of inflammatory biomarkers and GC-MS-based metabolite profiling.

Atriplex crassifolia (A. crassifolia) is a locally occurring member of Chenopodiaceae family that has been used in folk medicine for the treatment of joint pain and inflammation. The present study was focused to determine the analgesic and anti-inflammatory potential of the plant. n-hexane (ACNH) and methanol (ACM) extracts of A. crassifolia were evaluated for in vitro anti-inflammatory potential using protein denaturation inhibition assay. In vivo anti-inflammatory potential was determined by oral administration of 250, 500, and 1000mg/kg/day of extracts against carrageenan and formalin-induced paw edema models. Inflammatory mediators such as TNF-α, IL-10, IL-1β, NF-kB, IL-4, and IL-6 were estimated in blood samples of animals subjected to formalin model of inflammation. Analgesic activity was determined using acetic acid-induced writhing and tail flick assay model. Phytochemical profiling was done by GC-mass spectrophotometer. The results of in vitro anti-inflammatory activity revealed that both ACNH and ACM displayed eminent inhibition of protein denaturation in concentration-dependent manner. In acute in vivo carrageenan-induced paw edema model, both extracts reduced inflammation at 5th and 6th hour of study (p < 0.05). A. crassifolia extracts exhibited significant inhibition against formalin-induced inflammation with maximum effect at 1000mg/kg. ACNH and ACM significantly augmented the inflammatory mediators (p < 0.05). Levels of TNF-α, IL-6, IL-1β, and NF-kB were reduced, while those of IL-4 and IL-10 were upregulated. ACNH displayed maximum analgesic effect at 1000mg/kg, while ACM showed potent activity at 500 and 1000mg/kg. The extracts restored the CBC, TLC and CRP toward normal. GC-MS analysis revealed the presence of compounds like n-hexadecanoic acid, Phytol, (9E,11E)-octadecadienoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester, 1-hexacosene, vitamin E, campesterol, stigmasterol, gamma sitosterol in both extracts. These compounds have been reported to suppress inflammation by inhibiting inflammatory cytokines. The current study concludes that A. crassifolia possesses significant anti-nociceptive and anti-inflammatory potential owing to the presence of phytochemicals.

Immunohistochemical evidences of anticancer actions of metformin with other repurposed drug combinations and correlation with hamster fibrosarcoma tumor size

The aim was to detect and correlate anticancer effects of metformin in combinations with other repurposed drugs, already registered for other indications, which may be immediately applied and clinically investigated in oncology, reducing the time and cost of research for new cancer treatments. Immunohistochemistry was performed for tumors treated by dual drug combinations containing metformin with deoxycholic acid, caffeine, itraconazole, nitroglycerin, disulfiram or diclofenac. The drugs were applied in Syrian golden hamsters (6 animals per group) with the inoculated BHK21/C13 fibrosarcoma in doses equivalent to usual human doses, &lt;50 % LD50. The anticancer effects were assessed by: p53 (mutational status); Ki-67 and PCNA (tumor proliferation); CD34 and CD31 (neoangiogenesis); GLUT1 (glucose metabolism); iNOS (NO metabolism); COX4, Cytochrome C and caspase 3 (apoptosis); immunohistochemical markers. Also, biophysical characteristics of fibrosarcoma, animal blood samples and the toxicity on main organs were analyzed. Treatments significantly (P &lt; 0.05) reduced mutational status, tumor proliferation, neoangiogenesis, glucose metabolism, NO metabolism and modulated apoptosis, in correlation with tumor size, without toxicity and influence on biochemical blood and hematological tests. The administration of metformin in two-drug combination with deoxycholic acid, caffeine, itraconazole, nitroglycerin, disulfiram or diclofenac may be recommended for further clinical investigations in oncology.

First molecular detection of Francisella tularensis in turtle (Testudo graeca) and ticks (Hyalomma aegyptium) in Northwest of Iran

Francisella tularensis, causative agent of tularemia, is a contagious zoonotic ailment. This study was aimed to molecularly detect F. tularensis in tortoise blood (n = 100) and ticks (n = 100) collected in the West Azerbaijan province, Iran suing a 16SrRNA gene of the Francisella genus through employment of the Nested-PCR technique. The identified ticks were s Hyalomma aegyptium by morphological analysis. Seven percent (with a 95% CI: 3.5%–13.75%) of animal blood samples yielded positive results for the presence of the Francisella. Meanwhile, the Francisella was identified in tick samples at a rate of fifteen percent (15%) (with a 95% CI: 9%–23%). The samples containing positive results were specifically classified as F. tularensis subsp. holarctica. The samples were taken from ticks belonging to the H. aegyptium species that were gathered in Oshnavieh, southern part of West Azerbaijan province, Iran. This research was aimed to validate the existence of F. tularensis in ticks found within the West Azerbaijan province. Consequently, it is vital to acknowledge the potential of these ticks to transmit the bacteria to both livestock and humans through tick bites in this specific area.

Study of biotransformation of new selective carbonic anhydrase II inhibitor 4-(2-methyl-1,3-oxazole-5-yl)-benzenesulfonamide

The aim of the study was to determine biotransformation products of a new selective carbonic anhydrase II inhibitor – 4-(2-methyl-1,3-oxazole-5-yl)-benzenesulfonamide.Materials and methods. The study was conducted on 3 Wistar rats and 3 rabbits of the Soviet Chinchilla breed. The suspension of the drug was administered intraperitoneally to rats at a dosage of 20 mg/kg, to rabbits - at a dosage of 1.6 mg/kg. The animal blood samples were collected before the administration and 1, 2, 4, 24 h after. Urine sampling was also performed in the rats before the administration and in the intervals of 0–4, 4–8, 8–24 h after. The identification of metabolites in blood, urine and plasma was carried out using HPLC-MS/MS. Poroshell 120 C 18 column (50×3.0 mm, 2.7 µm) with a Zorbax Eclipse Plus C18 pre-column (12.5×2.1 mm, 5.0 µm) was used for the chromatographic separation. The assumed metabolites were synthesized, their structure was confirmed by the NMR spectroscopy method and a high-resolution mass spectrometry. The obtained substances were compared with the substances identified in biological fluids by retention time, the main MRM-transitions and mass spectra.Results. The N-hydroxymetabolite was revealed in the analyses of plasma, blood and urine samples which had been formed by the addition of an oxygen atom to the drug molecule. Chromatographic peaks of this compound were identified at the MRM-transitions of 255→159, 255→117, 255→89 m/z at the 7.2nd min of the analysis. The N-oxide of 4-(2-methyl-1,3-oxazole-5-yl)-benzenesulfonamide and N-hydroxy-4-(2-methyl-1,3-oxazole-5-yl)-benzenesulfonamide were synthesized; potentially, they could have been obtained during the biotransformation. During the confirmatory HPLC-MS/MS tests based on the coincidence of the retention times, the main MRM transitions and mass spectra, the ratio of the peak areas at the identified metabolite it was established that an N-hydroxy derivative. Chromatographic peaks of the N-oxide detected in the analysis of the model mixtures of the standard substance at the MRM-transitions of 255→175, 255→133, 255→89 m/z at the retention time of 5.43 min, were absent in the animal samples.Conclusion. The studied drug is metabolized to form a single metabolite of N-hydroxy-4-(2-methyl-1,3-oxazole-5-yl)-benzenesulfonamide. This compound was found in freshly collected samples of biological fluids of both animal species. The structure of the metabolite was confirmed by the HPLC-MS/MS-method by comparison with the synthesized standard substance.

Hydroxychloroquine Toxicity in the Vital Organs of the Body: In Vivo Study.

Hydroxychloroquine (HCQ) toxicity can adversely affect vital organs, cause pathologic ocular damage, and can have direct cardiovascular effects. This study aims to identify the biochemical, hematological, and histological alterations of the vital organs associated with the effects of HCQ. Male albino rats were exposed to the equivalent of HCQ therapeutic doses given to human patients being affected by malaria, lupus erythematosus, and COVID-19. The animal blood samples were subjected to hematological analysis, biochemical analysis, liver function tests, kidney function tests, and cardiac biomarkers. Liver, kidney, heart, spleen, and testis biopsies were subjected to histological examination. HCQ significantly lowered the values of erythrocytes, hemoglobin, hematocrit, platelets, leucocytes, and lymphocytes but significantly increased the values of aspartate aminotransferase (AST), alanine aminotransferase (ALT), amylase, alkaline phosphatase, lactate dehydrogenase, cholesterol, and chlorine ions. The renal tissues of HCQ-treated animals demonstrated glomerular fragmentation, partial atrophy degeneration, renal tubules hydropic degeneration, hyaline cast formation, and interstitial edema formation. Additionally, the heart exhibited myofiber necrosis, myolysis, wavy appearance, disorganization, and disarray. The testicular tissues also demonstrated spermatocyte degeneration, spermatogenic cell sloughing, testicular interstitial edema, and occasional spermatogenic arrest. Additionally, the spleen showed a decrease in the number and size of the white pulp follicles, a decrease in the number of apoptotic activity, and a decline in the number of T-rich cells. However, the red pulp demonstrated a diffuse decline in B rich-lymphocytes and macrophages. The liver was also the least affected but showed Kupffer cell hyperplasia and occasional hepatocyte dysplasia. The results indicate that chronic exposure to HCQ could alter the structures and functions of the vital organs.

Therapeutic mechanisms of the medicine and food homology formula Xiao-Ke-Yin on glucolipid metabolic dysfunction revealed by transcriptomics, metabolomics and microbiomics in mice

BackgroundIn recent decades, the prevalence of metabolic diseases, particularly diabetes, hyperlipidemia, obesity, and non-alcoholic fatty liver disease (NAFLD), has increased dramatically, causing great public health and economic burdens worldwide. Traditional Chinese medicine (TCM) serves as an effective therapeutic choice. Xiao-Ke-Yin (XKY) is a medicine and food homology TCM formula consisting of nine “medicine and food homology” herbs and is used to ameliorate metabolic diseases, such as insulin resistance, diabetes, hyperlipidemia and NAFLD. However, despite its therapeutic potential in metabolic disorders, the underlying mechanisms of this TCM remain unclear. This study aimed to evaluate the therapeutic effectiveness of XKY on glucolipid metabolism dysfunction and explore the potential mechanisms in db/db mice.MethodsTo verify the effects of XKY, db/db mice were treated with different concentrations of XKY (5.2, 2.6 and 1.3 g/kg/d) and metformin (0.2 g/kg/d, a hypoglycemic positive control) for 6 weeks, respectively. During this study, we detected the body weight (BW) and fasting blood glucose (FBG), oral glucose tolerance test (OGTT), insulin tolerance test (ITT), daily food intake and water intake. At the end of the animal experiment, blood samples, feces, liver and intestinal tissue of mice in all groups were collected. The potential mechanisms were investigated by using hepatic RNA sequencing, 16 S rRNA sequencing of the gut microbiota and metabolomics analysis.ResultsXKY efficiently mitigated hyperglycemia, IR, hyperlipidemia, inflammation and hepatic pathological injury in a dose dependent manner. Mechanistically, hepatic transcriptomic analysis showed that XKY treatment significantly reversed the upregulated cholesterol biosynthesis which was further confirmed by RT-qPCR. Additionally, XKY administration maintained intestinal epithelial homeostasis, modulated gut microbiota dysbiosis, and regulated its metabolites. In particular, XKY decreased secondary bile acid producing bacteria (Clostridia and Lachnospircaeae) and lowered fecal secondary bile acid (lithocholic acid (LCA) and deoxycholic acid (DCA)) levels to promote hepatic bile acid synthesis by inhibiting the LCA/DCA-FXR-FGF15 signalling pathway. Furthermore, XKY regulated amino acid metabolism including arginine biosynthesis, alanine, aspartate and glutamate metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, and tryptophan metabolism likely by increasing Bacilli, Lactobacillaceae and Lactobacillus, and decreasing Clostridia, Lachnospircaeae, Tannerellaceae and Parabacteroides abundances.ConclusionTaken together, our findings demonstrate that XKY is a promising “medicine food homology” formula for ameliorating glucolipid metabolism and reveal that the therapeutic effects of XKY may due to its downregulation of hepatic cholesterol biosynthesis and modulation of the dysbiosis of the gut microbiota and metabolites.

Abstract 971: Thymidine Kinase activity - A liquid biomarker reflecting proliferation rate and treatment efficacy in solid tumors provides unique guidance in oncology trials and drug development

Abstract Thymidine Kinase (TK) plays a fundamental role in DNA synthesis and cell proliferation. Expression of the enzyme is highly cell cycle dependent and correlates with cell growth state. TK activity (TKa) is present in all proliferating cells but release into blood circulation is low or absent from quiescent cells. Numerous studies have shown TKa to be elevated in solid tumors as well as in the blood of cancer patients with active disease. DiviTum® TKa, a clinically and analytically validated FDA cleared assay, can measure and quantify TKa in serum or plasma isolated from human and animal blood samples. TKa as a biomarker offers valuable information at all stages of drug development; from drug efficacy and dose response studies in cell culture and tumor xenograft models, to patient selection and treatment monitoring in clinical trials. Using TKa to measure the impact on cell proliferation as a drug evaluation endpoint can confirm mechanism of action efficacy and the minimally effective dose of a new compound rather than a maximally tolerated dose. In both cultured cells and mouse xenografts, a complete shutdown of tumor cell proliferation (assessed by TKa) can be observed well before and at much lower doses than when cell death or toxicity is used as the dose determination endpoint. The TKa-biomarker can also be used as a pharmacodynamic tool in clinical trials to improve study endpoints. Patient baseline levels (BL) of TKa are predictive/prognostic and can be used to identify which patients will respond to the therapy being tested vs those who will not. In metastatic breast cancer, a BL TKa value of &amp;lt;250 DuA is significantly correlated with longer PFS (HR=1.76, p&amp;lt;0.0001) and OS (HR=2.38, p&amp;lt;0.0001) independent of patient and disease characteristics. On-therapy TKa levels and dynamics can be used in clinical trials to evaluate response to a drug as early as two weeks into the first cycle. In patients receiving CDK4/6 inhibitors, TKa levels &amp;gt;140 DuA at cycle 1 day 15 predicts for lack of benefit to the drug (HR=5.65, p&amp;lt;0.0001). TKa can also be used as a predictive biomarker for drugs that cause an immune reaction and increase in cell proliferation such as immune checkpoint inhibitors (ICIs). When effective, ICIs cause an increase in immune cell proliferation which can be detected by elevated levels of TKa. Patients who do not achieve &amp;gt;1.5 fold change in TKa levels as compared to BL, have a significantly shorter duration of response to an ICI (HR=3.70, p=0.002) and patients with high TKa during ICI treatment have significantly longer OS (HR=3.49, p&amp;lt;0.05). In summary, TKa is a translational liquid biomarker that can bridge between preclinical and clinical studies, providing fundamental information for decision making during early and late stage drug development. Data and examples of the utility of TKa as a companion diagnostic for drug development will be presented. Citation Format: Mattias K. Bergqvist, Hanna Ritzén, Amy Williams. Thymidine Kinase activity - A liquid biomarker reflecting proliferation rate and treatment efficacy in solid tumors provides unique guidance in oncology trials and drug development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 971.

Identification and synthesis of metabolites of the new antiglaucoma drug

Introduction: The determination of biotransformation products is an essential part of the preclinical trial of original medicines. These studies have not been conducted before for the new drug 5-[5-(trifluoromethyl)-1,2-oxazole-3-yl]-furan-2-sulfonamide. Identification and synthesis of metabolite substances are necessary for subsequent tests of bioavailability, linearity of pharmacokinetics, accumulation, distribution and excretion. Materials and methods: The study was carried out on Wistar rats and rabbits of the Soviet Chinchilla breed. The substance of the drug was administered to animals intraperitoneally. The collection of animal blood samples was performed before administration and 1 h, 2 h, 4 h, 24 h after administration into K3EDTA-tubes. A part of each sample was centrifuged to separate the plasma. Rat urine samples were taken before administration and at intervals of 0-2 h, 2-4 h, 4-6 h, 6-24 h after administration of the drug. The HPLC-MS/MS method was used to identify metabolites in biological fluids. The assumed biotransformation products were synthesized after preliminary analysis. The structure of the obtained substances was confirmed by NMR spectroscopy and high-resolution mass spectrometry. Then, a comparison was carried out with the compounds identified in biological fluids by retention time, ratios of chromatographic peak areas at the main MRM-transitions using HPLC-MS/MS. Results and discussion: A metabolite formed by addition an oxygen atom to a drug molecule, as well as an acylated metabolite were detected during the analysis of plasma and blood samples. A compound with an increased molecular weight of 1 dalton compared to the drug substance was also present in a rat urine. N-hydroxy-5-[5-(trifluoromethyl)-1,2-oxazole-3-yl]-furan-2-sulfonamide, 5-[5-(trifluoromethyl)-1,2-oxazole-3-yl]-furan-2-sulfonic acid, which could potentially be obtained during biotransformation, as well as N-acetyl-5-[5-(trifluoromethyl)-1,2-oxazole-3-yl]-furan-2-sulfonamide were synthesized. Repeated HPLC-MS/MS tests confirmed the correctness of the initial hypothesis. It was found that a sulfonic acid derivative is formed in urine as a result of decomposition of N-hydroxymetabolite during sample collection. Conclusion: The studied drug is metabolized by formation of two metabolites: N-hydroxy-5-[5-(trifluoromethyl)-1,2-oxazole-3-yl]-furan-2-sulfonamide and N-acetyl-5-[5-(trifluoromethyl)-1,2-oxazole-3-yl]-furan-2-sulfonamide. N-hydroxymetabolite is able to decompose in biological fluids samples with formation of 5-[5-(trifluoromethyl)-1,2-oxazole-3-yl]-furan-2-sulfonic acid.

Potential miRNA biomarkers and therapeutic targets for early atherosclerotic lesions

Identification of potential therapeutic targets and biomarkers indicative of burden of early atherosclerosis that occur prior to advancement to life-threatening unstable plaques is the key to eradication of CAD prevalence and incidences. We challenged 16 baboons with a high cholesterol, high fat diet for 2 years and evaluated early-stage atherosclerotic lesions (fatty streaks, FS, and fibrous plaques, FP) in formalin-fixed common iliac arteries (CIA). We used small RNA sequencing to identify expressed miRNAs in CIA and in baseline blood samples of the same animals. We found 412 expressed miRNAs in CIA and 356 in blood samples. Eight miRNAs (miR-7975, -486-5p, -451a, -191-5p, -148a-3p, -17-5p, -378c, and -144-3p) were differentially expressed between paired fatty streak lesion and no-lesion sites of the tissue, and 27 miRNAs (e.g., miR-92a-3p, -5001, -342-3p, miR-28-3p, -21-5p, -221-3p, 146a-5p, and -16-5p) in fibrous plaques. The expression of 14 blood miRNAs significantly correlated with extent of lesions and the number of plaques. We identified coordinately regulated miRNA-gene networks in which miR-17-5p and miR-146a-5p are central hubs and miR-5001 and miR-7975 are potentially novel miRNAs associated with early atherosclerosis. In summary, we have identified miRNAs expressed in lesions and in blood that correlate with lesion burden and are potential therapeutic targets and biomarkers. These findings are a first step in elucidating miRNA regulated molecular mechanisms that underlie early atherosclerosis in a baboon model, enabling translation of our findings to humans.

The discovery of human Plasmodium among domestic animals in West Sumba and Fakfak, Indonesia

Background: In Indonesia, malaria incidence is at a high rate despite maximum preventive efforts. Therefore, this study aims to determine the possibility of a Plasmodium reservoir among domestic animals in malaria-endemic areas. Methods: Animal blood was collected using EDTA tubes, then smeared and stained with Giemsa for Plasmodium microscopic identification. About 10 µl of blood was dropped on to a filter paper to capture Plasmodium DNA. Nested PCR was used for parasite molecular detection, while Plasmodium species were identified using the sequenced DNA. Results: A total of 208 and 62 animal blood samples were collected from Gaura village, West Sumba and Fakfak village, West Papua, Indonesia respectively. In total, 32 samples from Gaura contained P. falciparum or P. vivax, while the Plasmodium percentage in buffalo, horse, goat, and dogs were 20.7%, 14.3%, 5.8%, 16.7%, respectively. P. knowlesi was not found in any of the samples, and no other species were detected in 18 pig blood samples. Conclusion: Human Plasmodium existence among domestic animals in Indonesia partly explains the high prevalence and persistence of malaria in some endemic areas due to a reservoir host presence. Therefore, future studies need to ascertain the cause.

ГЕМАТОБИОХИМИЧЕСКИЙ СТАТУС СОБАК ПРИ ИСПОЛЬЗОВАНИИ БИОКОМПОЗИЦИОННОГО ПОКРЫТИЯ ИМПЛАНТОВ ДЛЯ УСКОРЕНИЯ КОНСОЛИДАЦИИ КОСТНОЙ ТКАНИ

Injuries of the musculoskeletal system of dogs occur in more than 30% of the total number of non-infectious pathologies. In veterinary and humane traumatology, the problems associated with fractures and their complications have not been fully resolved. Due to the use of different methods of osteosynthesis using different implants (spokes, bone plates, osteofixers, pins), it is quite important to apply coatings for these materials that met such specified needs as: thickness, morphological heterogeneity, roughness, porosity, adhesive strength and antibacterial effect to accelerate the consolidation processes. The article shows the results of studies on the dynamics of hematological and biochemical parameters of groups of experimental animals (dogs), in view of the use of osteoplastic biocomposition coating for implants (spokes) in intramedullary and extra-focal osteosynthesis of tubular bones. Animal blood samples (n=120) served as the material for research. The purpose of this study was to assess the dynamics of hematological and biochemical parameters of blood during the implantation of spokes with the developed osteoplastic biocomposition coating. A group of authors, based on the dynamics of hematological parameters, determined that the coating developed by them, consisting of hydroxyapatite, methyluracil, amoxicillin and polylactide, does not cause hematopoiesis inhibition and acute inflammatory reaction. Based on the dynamics of biochemical parameters, it was determined that this coating does not have hepatotoxicity and nephrotoxicity, the absence of a negative effect of this material on the process of bone restoration and the formation of a callus was determined. The developed coating, from the point of view of the authors, allows for a sufficiently high degree and speed of consolidation of bone fragments with the prevention of bacterial contamination.

Avaliação de risco ecológico em potencial avaliação de metal tóxico (chumbo) em prados contaminados nas proximidades da cidade suburbana: solo x forragens x gado

Heavy metal toxicity is becoming an increasing concern for environmental, human and animal health. The current research analyzed the lead (Pb) contamination in the food chain under three different irrigation sources (ground, canal, and wastewater). Soil, plant and animal samples were collected from the Jhang district of Pakistan and processed with an atomic absorption spectrophotometer. Lead concentration varied in the samples as: 5.22-10.73 mg/kg in soil, 2.46-10.34 mg/kg in forages and 0.736-2.45 mg/kg in animal samples. The observed lead concentration in forage and animal blood samples was higher than the standard limits. The pollution load index (0.640-1.32) in soil showed that lead contamination mainly took place at the wastewater irrigating sites. Bio-concentration factor values (0.313-1.15) were lower than one in all samples except Zea mays, showing that lead metal was actively taken up by Zea mays tissues from the soil. Enrichment factor values ranged from 0.849-3.12, showing a moderate level of lead enrichment. Daily intake and health risk index varied between 0.004-0.020 mg/kg/day and 0.906-4.99, respectively. All the samples showed maximum lead concentration at the wastewater irrigating site compared to the ground or canal water application sites. These results recommended that consistent application of wastewater for forage irrigation must be avoided to prevent health hazards associated with lead in the animal and human food chain. Government must implement adequate strategies to protect the animal and human health from the harms of toxic heavy metals.

Classification of bloodstains deposited at different times on floor tiles using hierarchical modelling and a handheld NIR spectrometer.

Bloodstains are commonly encountered at crime scenes, especially on floor tiles, and can be deposited over different periods and intervals. Therefore, it is crucial to develop techniques that can accurately identify bloodstains deposited at different times. This study builds upon a previous investigation and aims to enhance the performance of three distinct hierarchical models (HMs) designed to differentiate and identify stains of human blood (HB), animal blood (AB), and common false positives (CFPs) on nine different types of floor tiles. Soft Independent Modeling Class Analogies (SIMCA), and Partial Least Squares-Discriminant Analysis (PLS-DA) were employed as decision rules in this process. The originally published model was constructed using a training set that included samples with a known time of deposit of six days. This model was then tested to predict samples with various deposition times, including human blood samples aged for 0, 1, 9, 20, 30, and 162 days, as well as animal blood samples aged for 0, 1, 10, 13, 20, 29, 105, and 176 days. To improve the identification of human blood, the models were modified by adding zero-day and one-day-old bloodstains to the original training set. All models showed improvement when fresher samples were included in the training set. The best results were achieved with the hierarchical model that used partial least squares-discriminant analysis as the second decision rule and incorporated one-day-old samples in the training set. This model yielded sensitivity values above 0.92 and specificity values above 0.7 for samples aged between zero and 30 days.