Cat Plasma Research Articles

Development of a UPLC-MS/MS method for quantifying KPT-335 (Verdinexor) in feline plasma for a study of PK.

KPT-335 (Verdinexor) is a novel SINE that potently inhibits the nucleoprotein Exportin 1 (XPO1/CRM1) of tumor cell lines and reduces the replication level of the influenza virus. KPT-335 is mainly used for the treatment of canine tumors. Drugs for the effective treatment of feline tumors are currently unavailable in China. KPT-335 may have potential in the treatment of cat tumors. However, the effects of KPT-335 in cats are unreported, and no relevant methodology has been established for pharmacokinetic studies. In this study, a UPLC-MS/MS method was developed to determine KPT-335 concentrations in cat plasma, followed by pharmacokinetic studies. Briefly, plasma proteins are precipitated with acetonitrile, and the supernatant was collected for detection after centrifugation. The linearity for KPT-335 in cat plasma was in the range of 5-1,000 ng/mL. Satisfactory accuracy and precision were obtained. The intra-day accuracy was between -4.10% and 10.48%, the precision was ≤4.65%; the inter-day accuracy was between -0.11% and 8.09%, and the precision was ≤5.85%. Intra-day and inter-day accuracy and precision were within regulatory limits. The results of preliminary pharmacokinetic studies were as follows: Tmax was 1.46 ± 0.51 h; Cmax was 239.54 ± 190.60 ng·mL-1; T1/2 was 5.16 ± 2.30 h; AUC0-t was 1439.85 ± 964.64 ng·mL-1·h. The AUC0-∞ was 1589.82 ± 1003.75 ng·mL-1·h. The purpose of this study was to develop a rapid and simple UPLC-MS/MS method to detect KPT-335 concentration in cat plasma and to conduct preliminary pharmacokinetic studies to support the future application of KPT-335 in felines.

Development and validation of an LC-MS/MS method for the quantification of oral-sugar probes in plasma to test small intestinal permeability and absorptive capacity in the domestic cat (Felis catus)

A novel method for quantifying the concentration of lactulose, rhamnose, xylose, and 3-O-methylglucose (3-OMG) in cat plasma using liquid chromatography-mass spectrometry (LC-MS) was developed. Domestic male cats (n = 13) were orally dosed with a solution containing the four sugars to test the permeability and absorptive capacity of their intestinal barrier. Plasma samples were taken 3 h later and were prepared with acetonitrile (ACN), dried under N2, and reconstituted in 90 % ACN with 1 mM ammonium formate. Stable isotope labelled 13C standards for each analyte were used as internal standards. Chromatographic separation was conducted using a Phenomenex Luna NH2 column with a gradient elution system of deionized water and 90 % ACN with 1 mM ammonium formate at 300 µL/min for 13 min total analysis time. Recovery trials were conducted in triplicate over three days with RSD values (%) for each day ranging from 1.2 to 1.4 for lactulose, 5.4 – 6.0 for rhamnose, 3.3 – 5.5 for xylose, and 2.6 – 5.6 for 3-OMG. Inter-day variations for each analyte were not different (p > 0.05). Limit of detection and quantification were 0.2 and 0.7 µg/mL for lactulose, 0.8 and 2.4 µg/mL for rhamnose, 0.6 and 1.8 µg/mL for xylose, and 0.3 and 1.1 µg/mL for 3-OMG, respectively. Plasma sugar concentrations recovered from cats were above the limit of quantification and below the highest calibration standard, validating the use of this method to test intestinal permeability and absorptive capacity in cats.

Detection of Protein Biomarkers Relevant to Sperm Characteristics and Fertility in Semen in Three Wild Felidae: The Flat-Headed Cat (Prionailurus planiceps), Fishing Cat (Prionailurus viverrinus), and Asiatic Golden Cat (Catopuma temminckii).

Effective wild cat conservation programs with assisted reproductive technologies are being developed in different parts of the world. The flat-headed cat, fishing cat, and Asiatic golden cat are three species among nine wild Felidae in Thailand that are in need of urgent conservation efforts. Here, we assessed routine sperm characteristics and we report the detection of protein biomarkers related to the fertilization process, IZUMO1 and the CRISP family, and apoptotic markers, active or cleaved caspase-3, in semen samples collected from these wild cats. IZUMO1 was located in the equatorial segment of the sperm head, which is the region involved in gamete interaction. The highest levels of IZUMO1 were found in both the sperm pellet and the seminal plasma of the flat-headed cat, as determined by immunoblotting. CRISP2, a sperm-egg fusion assisting protein, and CRISP3 were found in both the sperm pellet and the seminal plasma, and the highest levels were observed in the fishing cat. Positive correlations between certain semen parameters and IZUMO1, CRISP2, and CRISP3 expression were also demonstrated. Cleaved caspase-3 was found in all sperm samples in all three species and was associated with an increase in DNA fragmentation and a decrease in certain semen characteristics such as motility, viability, and intact acrosomes. Our results suggest that the analysis of IZUMO1, the CRISP family, and cleaved caspase-3, along with the routine sperm characteristics, may allow for better success in breeding management in wild Felidae, particularly in the flat-headed cat and the fishing cat.

An Aptamer-Based Proteomic Analysis of Plasma from Cats (Felis catus) with Clinical Feline Infectious Peritonitis.

Feline infectious peritonitis (FIP) is a systemic disease manifestation of feline coronavirus (FCoV) and is the most important cause of infectious disease-related deaths in domestic cats. FIP has a variable clinical manifestation but is most often characterized by widespread vasculitis with visceral involvement and/or neurological disease that is typically fatal in the absence of antiviral therapy. Using an aptamer-based proteomics assay, we analyzed the plasma protein profiles of cats who were naturally infected with FIP (n = 19) in comparison to the plasma protein profiles of cats who were clinically healthy and negative for FCoV (n = 17) and cats who were positive for the enteric form of FCoV (n = 9). We identified 442 proteins that were significantly differentiable; in total, 219 increased and 223 decreased in FIP plasma versus clinically healthy cat plasma. Pathway enrichment and associated analyses showed that differentiable proteins were related to immune system processes, including the innate immune response, cytokine signaling, and antigen presentation, as well as apoptosis and vascular integrity. The relevance of these findings is discussed in the context of previous studies. While these results have the potential to inform diagnostic, therapeutic, and preventative investigations, they represent only a first step, and will require further validation.

Characterization of platelet rich plasma in feline immunodeficiency virus-infected cats: Cell, and PDGF-BB and TGF-ß1 growth factor analysis

Autologous platelet-rich plasma (PRP) contains growth factors (GFs) that modulate the expression of inflammatory cells; thus, these products could be considered a good strategy to favor tissue regeneration in feline immunodeficiency (FIV) positive cats. However, there is no scientific documentation on obtaining PRP in FIV-positive cats.Authors hypothesized that PRP can be obtained in FIV cats following the PRGF®-Endoret® methodology. The objectives of this study were to compare the platelet, erythrocyte, and leukocyte concentration between whole blood (WB) and the PRP; and determine the concentration of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor β1 (TGF-β1) in FIV-positive cats. Sixteen adults FIV-positive asymptomatic cats were included in the study. WB samples were drawn and the PRP was obtained by centrifugation at 265g for 10 min. Erythrocyte and leukocyte, platelets, and mean platelet volume (MPV) were determined both in WB and in PRP. PDGF-BB and TGF-β1 concentrations were additionally determined in PRP.Platelet concentration increased 1.1 times in PRP fraction compared to WB, but no significant differences were reported. MPV was statistically higher in WB than in PRP (p = 0.001). Erythrocytes and leukocytes counts were decreased by 99% and 92%, respectively in the PRP fraction (p < 0.001). Regarding TGF-ß1, a higher concentration was shown in the PRP (p < 0.02). Although the product obtained could not be classified as PRP according to the PRGF®-Endoret® methodology, based on the drastic reduction of RBC and WBC, the PLT concentrate is of high purity.

The effects of a hydrolyzed protein diet on the plasma, fecal and urine metabolome in cats with chronic enteropathy

Hydrolyzed protein diets are extensively used to treat chronic enteropathy (CE) in cats. However, the biochemical effects of such a diet on feline CE have not been characterized. In this study an untargeted 1H nuclear magnetic resonance spectroscopy-based metabolomic approach was used to compare the urinary, plasma, and fecal metabolic phenotypes of cats with CE to control cats with no gastrointestinal signs recruited at the Royal Veterinary College (RVC). In addition, the biomolecular consequences of a hydrolyzed protein diet in cats with CE was also separately determined in cats recruited from the RVC (n = 16) and the University of Bristol (n = 24) and whether these responses differed between dietary responders and non-responders. Here, plasma metabolites related to energy and amino acid metabolism significantly varied between CE and control cats in the RVC cohort. The hydrolyzed protein diet modulated the urinary metabolome of cats with CE (p = 0.005) in both the RVC and Bristol cohort. In the RVC cohort, the urinary excretion of phenylacetylglutamine, p-cresyl-sulfate, creatinine and taurine at diagnosis was predictive of dietary response (p = 0.025) although this was not observed in the Bristol cohort. Conversely, in the Bristol cohort plasma betaine, glycerol, glutamine and alanine at diagnosis was predictive of outcome (p = 0.001), but these same results were not observed in the RVC cohort. The biochemical signature of feline CE in the RVC cohort was consistent with that identified in human and animal models of inflammatory bowel disease. The hydrolyzed protein diet had the same effect on the urinary metabolome of cats with CE at both sites. However, biomarkers that were predictive of dietary response at diagnosis differed between the 2 sites. This may be due to differences in disease severity, disease heterogeneity, factors unrelated to the disease or small sample size at both sites. As such, further studies utilizing larger number of cats are needed to corroborate these findings.

Immunoreactivity of canine, feline, and equine D-dimer with antibodies to human D-dimer.

Commercially available D-dimer assays use antibodies against human D-dimer, with limited sensitivity and specificity data in companion animals. To evaluate the immunoreactivity of D-dimer in plasma of dogs, horses, and cats with commercially available antibodies to human D-dimer. Plasma samples were collected from healthy dogs and horses, and from surplus feline plasma submitted for diagnostic purposes. Descriptive research study. A cross-linked fibrin lysate was prepared from plasma samples, and SDS-PAGE and immunoblotting were performed with a variety of commercially available antibodies to human D-dimer. The selected antibodies demonstrated variable reactivity with D-dimer of each species. The monoclonal antibody DD44 bound canine D-dimer with good specificity and sensitivity, but this antibody did not react with feline or equine D-dimer. The polyclonal antibody D2D bound putative D-dimer in dogs, cats, and horses with good specificity, and higher sensitivity compared to human D-dimer. The variable performance of commercially available human D-dimer assays between species is, in part, because of inter-species variation in D-dimer immunoreactivity. The use of these assays should follow validation studies. Monoclonal antibody DD44 could be a focus for the development of a canine-specific assay.

Development and validation of an LC-MS/MS method for the simultaneous quantification of milbemycin oxime and praziquantel in plasma: application to a pharmacokinetic study in cats.

Milbemycin oxime (MBO) and praziquantel (PZQ) have a broad spectrum of biological activity and are commonly used to treat the parasitic infection in the veterinary clinic. In this study, a fast and efficient LC-MS/MS method was established and validated for the simultaneous determination of MBO, PZQ, cis-4-hydroxylated-PZQ (C-4-OH-PZQ) and trans-4-hydroxylated-PZQ (T-4-OH-PZQ) and in cat plasma. Extraction of analytes and internal standards from cat plasma by acetonitrile protein precipitation, allows rapid processing of large batches of samples. MBO, PZQ, C-4-OH-PZQ, T-4-OH-PZQ, and internal standard (IS) were eluted for 13.5 min on a C18 column with a 0.1% formic acid water/acetonitrile mixture as the mobile phase. Results showed that the method had good precision, accuracy, recovery, and linearity. The linearity range was 2.5-250 ng/mL for MBO, and 10-1000 ng/mL for PZQ, C-4-OH-PZQ, and T-4-OH-PZQ. The intra-day and inter-day precision CV values of the tested components were within 15%. The extraction recoveries of the four components ranged from 98.09% to 107.46%. The analytes in plasma remained stable for 6 h at room temperature, 26 h in the autosampler (4 °C), after freeze-thaw (-20°C) cycles, and 60 days in a -20°C freezer. Method sensitivity sufficed for assessing pharmacokinetic parameters of MBO, PZQ, C-4-OH-PZQ, and T-4-OH-PZQ in plasma samples with LLOQ of 2.5 ng/mL for MBO and 10 ng/mL for PZQ, C-4-OH-PZQ, and T-4-OH-PZQ. In this study, a selective and sensitive LC-MS/MS method for the simultaneous quantification of MBO, PZQ, C-4-OH-PZQ, and T-4-OH-PZQ in cat plasma was developed and validated.This method had been successfully applied to evaluate the pharmacokinetics of MBO, PZQ, C-4-OH-PZQ, and T-4-OH-PZQ after a single oral administration of 8 mg MBO and 20 mg PZQ in cats.

Population pharmacokinetic analysis of enrofloxacin and its active metabolite ciprofloxacin after intravenous injection to cats with reduced kidney function.

It is unknown if enrofloxacin accumulates in plasma of cats with reduced kidney function. To determine if enrofloxacin and its active metabolite ciprofloxacin have reduced clearance in azotemic cats. Thirty-four cats hospitalized for clinical illness with variable degree of kidney function. Prospective study. After enrofloxacin (dose 5 mg/kg) administration to cats, sparse blood sampling was used to obtain 2 compartment population pharmacokinetic results using nonlinear mixed-effects modeling. Plasma enrofloxacin and ciprofloxacin concentrations were measured and summed to obtain the total fluoroquinolone concentration. A model of ciprofloxacin metabolism from enrofloxacin was created and evaluated for covariate effects on clearance, volume of distribution, and the metabolic rate of ciprofloxacin generation from enrofloxacin. Body weight was the only covariate found to affect total fluoroquinolone volume of distribution (effect 1.63, SE 0.19, P < .01) and clearance (effect 1.63, SE 0.27, P < .01). Kidney function did not have a significant effect on total fluoroquinolone clearance (median 440.8 mL/kg/h (range 191.4-538.0 mL/kg/h) in cats with normal kidney function, 365.8 mL/kg/h (range 89.49-1092.0 mL/kg/h) in cats with moderate kidney dysfunction, and 308.5 mL/kg/h (range 140.20-480.0 mL/kg/h) in cats with severe kidney dysfunction (P = .64). Blood urea nitrogen concentration influenced the metabolic generation of ciprofloxacin from enrofloxacin (effect 0.51, SE 0.08, P < .01), but other markers of kidney function did not. Adjustment of enrofloxacin dosage is not indicated for azotemic cats.

Abundance of Anti-Muellerian hormone in cat ovaries and correlation of its plasma concentration with animal age, weight and stage of the estrous cycle

In female animals of different species, Anti-Müllerian hormone (AMH) is produced by follicular granulosa cells and has been associated with the ovarian follicle pool. Because concentration of AMH in plasma of ovary-intact female cats is apparently more variable than previously assumed, we have analysed AMH concentration in blood of cats (n = 93) presented for routine ovariectomy and assessed ovarian histology and AMH protein expression in the surgically removed ovaries. We hypothesised that AMH is synthesized only in preantral and small antral follicles and that plasma AMH concentration reflects the antral follicle count (AFC). Corpora lutea were detected in 35% of the female cats, whereas plasma progesterone concentration was ≥1 ng/mL in 57% of the cats. Follicular cysts were present in 15 cats (16%). Positive immunostaining for AMH protein was detected in close to all primordial and antral follicles, ovarian cysts, 70% of corpora lutea and 28% of atretic follicles. Concentration of AMH in plasma averaged 6.8 ± 0.5 ng/mL (range 1.3–21.7 ng/mL). The AFC increased with increasing AMH concentration with a moderate positive correlation between AFC and AMH (r = 0.286, p < 0.01). Plasma AMH concentration was not affected by season or cats’ age, weight, stage of the estrous cycle and presence of follicular cysts. In conclusion, AMH protein is expressed in all endocrine structures of the cat ovary. While AMH is a marker for the presence of ovarian tissue, its usefulness to assess ovarian function in individual female cats is of limited value.

Plasma metabolomics identifies metabolic alterations associated with the growth and development of cat.

The purpose of our study was to study the composition and content of the feline plasma metabolome revealing the critical metabolites and metabolic pathways associated with age during growth and development. Blood samples were collected from juvenile and adult groups for blood routine tests and serum biochemistry tests. Non-targeted metabolomics analyses of plasma were also performed to investigate changes in metabolites and metabolic pathways. In this study, we found that the red blood cell counts, liver function indexes (albumin and gamma-glutamyl transpeptidase), and the concentration of triglyceride and glucose changed significant with growth and development. The metabolomics results revealed that 1427 metabolites were identified in the plasma of young and adult cats. Most of these metabolites belong to major classes of lipids and lipid-like molecules. The most obvious age-related metabolites include reduced levels of chenodeoxycholate, taurocholate, cholate, and taurochenodeoxycholate but increased levels of L-cysteine and taurocyamine in the adult cat's serum. These metabolites are mainly involved in the primary bile acid biosynthesis pathway, the bile secretion pathway, and the taurine and hypotaurine metabolism pathway. This study revealed many age-related metabolite alterations in the feline plasma. These age-varying metabolites, especially in the bile acid biosynthesis and secretion metabolism pathways, indicate that the regulation of these pathways is involved in the growth and development of cats. This study promotes our understanding of the mechanism of feline growth and provides new insights into nutrition and medicine for cats of different ages.

8-Hydroxy-2'-deoxyguanosine and malondialdehyde in plasma and their association with disease severity in 20 cats with chronic kidney disease.

Oxidative stress is associated with the development and progression of chronic kidney disease (CKD) in humans. The aim of this study was to evaluate the concentrations of oxidative stress markers, 8-hydroxy-2'-deoxyguanosine (8-OHdG) and malondialdehyde (MDA), in the plasma and urine of cats with different stages of CKD. Plasma and urine samples were collected from cats with CKD that were referred to the Veterinary Medical Center of the University of Tokyo between April 2019 and October 2022. Plasma and urine samples were collected from healthy cats (n = 6 at most), cats with stage 2 CKD (n = 8) and stage 3-4 CKD (n = 12), and cats with idiopathic cystitis (disease control, n = 5). Plasma and urine concentrations of 8-OHdG and MDA were measured using ELISA and thiobarbituric acid reactive substances assay kits, respectively. The median plasma 8-OHdG concentrations were 0.156 ng/ml (<0.125-0.210 ng/ml) in the healthy group, <0.125 ng/ml (range <0.125 ng/ml) in the idiopathic cystitis group, 0.246 ng/ml (range 0.170-0.403 ng/ml) in cats with stage 2 CKD and 0.433 ng/ml (range 0.209-1.052 ng/ml) in cats with stage 3-4 CKD. Concentrations in stage 3-4 CKD were significantly higher than those in the healthy and disease control groups. Plasma MDA concentrations were low in the healthy and disease control groups and significantly higher in cats with stage 3-4 CKD. In every cat with CKD, plasma 8-OHdG and MDA concentrations were positively correlated with plasma creatinine concentrations (8-OHdG, rs = 0.68; MDA, rs = 0.67). Urinary 8-OHdG/urinary creatinine (u-CRE) and urinary MDA/u-CRE levels did not differ significantly between the groups; however, it was difficult to evaluate them because of the small sample size. This report shows that plasma 8-OHdG and MDA concentrations increase with the severity of feline CKD. These markers may be useful for assessing oxidative stress in cats with CKD.

Determination of blood heavy metal concentrations and oxidant-antioxidant capacities in Angora cats at different age and gender

This study was conducted to investigate heavy metal concentrations and oxidative status of plasma and erythrocytes in Angora cats at different ages and gender. Sixteen young (less than 1 year old) and 14 adult (1–6 years old) cats were also grouped according to gender as male (n = 17) and female (n = 13). The separated plasma samples from cat’s blood were analyzed for selected heavy metals and total oxidant and antioxidant capacities (TOC and TAC) and calculated for oxidative stress index (OSI). The erythrocyte hemolysates were also evaluated for malondialdehyde (MDA), and super oxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX). Results of the study showed that most of the measured metals were not varied statistically according to age or gender. However, adult cats had significantly (P&amp;lt;0.01) higher Cu and lower Fe levels compared to young cats. Plasma levels of TOC, TAC and OSI, and erythrocyte MDA concentrations in young cats were significantly (P&amp;lt;0.05) higher than that of adults. While the SOD activity was decreased by the age, GPX activity was increased (P&amp;lt;0.05). However, the activity of CAT was changed by only gender, which was higher in males (P&amp;lt;0.01). In conclusion, metals, especially trace elements, are required for many kinds of physiological processes and the synthesis of antioxidant enzymes. Therefore, it can be suggested that the periodic measurement of metals and the addition of common antioxidant supplements to the diet of adult Angora cats will support weakening antioxidant mechanisms by age.

Diets Causing Taurine Depletion in Cats Substantially Elevate Postprandial Plasma Cholecystokinin Concentration

Excessive secretion of the intestinal hormone cholecystokinin (CCK) was postulated to cause diet-related taurine depletion in cats. To test this hypothesis, plasma CCK-like immunoreactivity (CCK-LI) was measured in cats given four diets, two purified and two canned, that contained similar concentrations of protein, fat, moisture and taurine but produced variable rates of taurine depletion. Plasma CCK-LI was measured by RIA with a tyrosine-sulfate specific, C-terminal antiserum, validated for use in cat plasma. As indicated by measurements of taurine in whole blood and urine, a purified diet containing casein maintained body taurine, whereas the same diet containing soybean protein and a commercial canned diet preserved either by freezing or cooking depleted body taurine. Preprandial and peak postprandial plasma CCK-LI in cats given the casein-containing diet were 10.6 ± 1.4 and 27.6 ± 4.8 pmol/L, respectively, ∼two- to tenfold greater than those reported in humans. Integrated postprandial plasma CCK-LI was less for cats given the casein diet than cats given both forms of the canned diet; it tended to be lower in cats given the casein diet than in cats given the soy protein diet. A negative linear correlation was observed between apparent nitrogen digestibilities of the diets and integrated plasma CCK-LI. The results indicated that diets that cause taurine depletion have lower protein digestibilities and cause greater endocrine secretion of CCK than diets that maintain body taurine status.

Dietary Soybean Protein Decreases Plasma Taurine in Cats

Commercial dry and canned diets fed to cats cause ∼two- and fourfold increase in the taurine requirement, respectively, as compared with that observed for purified diets. In two experiments, the effect of source and level of protein and fiber in the diet on the concentration of taurine in plasma and whole blood of cats was studied. All diets contained 1 g taurine/kg dry matter. When a casein-based diet containing either 25% or 50% protein was given to cats for 6 wk, no difference in plasma taurine concentration was observed; however, substituting soybean protein for casein resulted in a significant (P < 0.01) decrease in plasma taurine concentration of cats in the 50% soybean protein group, but not in the 25% soybean protein group. In Experiment 2, the food intake of cats was limited [26 g dry matter/(kg body weight·d)], and the protein was 30 or 60% of the diet. Cats fed 60% soybean protein or casein diets had significantly lower plasma taurine concentrations than cats fed a 30% casein diet, with the 60% soybean protein diet causing the greater decrease. There was no effect of either 2 or 4% soybean fiber on plasma taurine concentration as compared with the same diet without the added fiber. The taurine concentration in plasma was higher (P < 0.05) in male cats than in female cats. Protein source, amount in the diet and gender did not affect the whole blood taurine concentration. Cats given diets containing 60% casein or soybean protein diets excreted a greater amount of fecal total bile acid and total taurine than cats given a 30% casein diet. Cats with higher plasma concentrations of taurine excreted a greater amount of free taurine in urine, and a lesser amount of taurine and bile acids in feces. These results show that although protein source (soybean protein) and the quantity in the diet have a significant effect on the excretion pattern of taurine by cats, these effects are not sufficient to account for the marked increase in the taurine requirement found when canned heat-processed diets are fed.

Research of the New Biomarkers of Mammary Gland Tumors in Domestic Cats

This article presents the results of the clinical significance study of changes in the activity of adenosine deaminase and the content of polyamines in the blood plasma of cats with mammary gland tumors. In our work, we investigated the activity of the adenosine deaminase gene in the tissues of mammary gland tumors in cats and compared the activity of this gene with normal mammary gland tissue. In the course of this work, we also found out the frequency of occurrence of mammary gland tumors in the population of domestic cats in the Rostov region and evaluated the prognosis of survival after surgical treatment of breast cancer. The incidence of mammary gland tumors was detected, which amounted to 0.4% of the domestic cats who have applied to veterinary clinics in Rostov region, where the cats with mammary gland neoplasms amounted to approximately 400 individuals for every 100,000 animals. These data reflect the importance of the problem of mammary gland tumors and emphasize the need to study the pathophysiological foundations of carcinogenesis in order to create molecular genetic approaches for early diagnosis of mammary gland tumors. The results of the studies showed that the level of expression of the adenosine deaminase gene in the tissues of invasive nonspecific carcinoma and in the tissues of fibroepithelial hyperplasia sharply increases compared to the tissue of a healthy mammary gland, 36 times in the tissues of invasive carcinomas and 131 times in the tissues of fibroepithelial hyperplasia. At the same time, there were no significant differences between the activity of adenosine deaminase in the blood plasma of healthy animals and in the blood plasma of animals with malignant neoplasms of the mammary gland. Measurement of the level of polyamines in the blood of animals showed that the content of putrescine in the erythrocytes of the blood of cats with benign and tumor-like neoplasms of the mammary fibroepithelial hyperplasia significantly increased by 5 times compared with the indicators of the control group. In malignant neoplasms of the mammary gland invasive nonspecific carcinoma, the content of putrescine and spermin in the blood significantly exceeded the control values by 6 and 10 times, respectively.

The Relationship of Clinicopathological Parameters and Formula for Predicting Malondialdehyde Level in Feline Immunodeficiency Virus (FIV) - infected Cats

The objective of this study was to investigate clinicopathological parameters, malondialdehyde and total antioxidant power for analyzing correlation and predicting malondialdehyde in plasma of feline immunodeficiency virus (FIV)-infected cats. Body condition score (BCS), mucous membrane color score (MMCS), vital signs, hematological parameters, malondialdehyde and ferric reducing antioxidant power (FRAP) of 20 FIV-infected cats were examined. The correlation and multiple regression model were then analyzed. The results revealed the following. The correlation between hemoglobin vs. packed cell volume (PCV), total red blood cell (TRBC) vs. PCV, TRBC vs. hemoglobin, mean corpuscular hemoglobin (MCH) vs. mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC) vs. MCH, body temperature vs. eosinophil, total white blood cell (TWBC) vs. eosinophil, MMCS vs. platelet, eosinophil vs. malondialdehyde, PCV vs. FRAP, hemoglobin vs. FRAP, TRBC vs. FRAP and TRBC vs. FRAP were significantly positive correlated between pairs of these parameters (P&lt;0.05), while body temperature vs. MCV, TRBC vs. MCV, heart rate vs. platelet, heart rate vs. platelet were significantly negatively correlated between pairs of parameters (P&lt;0.05). A formula for predicting malondialdehyde was Heart Rate (beats/minute) = 166.341 + 5.438*Hemoglobin (g/dl) + 0.0009*Total White Blood Cell (cell/ml) - 1152.48*Malondialdehyde (µmol/ml) (MSE=132.2540; r2=0.85). These results indicated that there were correlations between some clinicopathological parameters, malondialdehyde and total antioxidant power and a formula for predicting malondialdehyde levels in plasma of FIV-infected cat was obtained.

Identification of potential plasma protein biomarkers for feline pancreatic carcinoma by liquid chromatography tandem mass spectrometry.

In both humans and cats, pancreatic carcinoma is an aggressive cancer with a grave prognosis. Proteomics techniques have successfully identified several blood-based biomarkers of human pancreatic neoplasia. Thus, this study aims to investigate whether similar biomarkers can be identified in the plasma of cats with FePAC by using liquid chromatography tandem mass spectrometry (LC-MS/MS). To facilitate evaluation of the low abundance plasma proteome, a human-based immunodepletion device (MARS-2) was first validated for use with feline plasma. Marked reduction and/or complete removal of albumin and immunoglobulins was confirmed by analysis of electrophoretograms and mass spectral data. Subsequently, plasma collected from 9 cats with pancreatic carcinoma (FePAC), 10 cats with symptomatic pancreatitis, and 10 healthy control cats was immunodepleted and subjected to LC-MS/MS. Thirty-seven plasma proteins were found to be differentially expressed (p < .05 in one-way ANOVA, FC >2 in fold change analysis). Among these proteins, ETS variant transcription factor 4 (p < .05) was overexpressed, while gelsolin (p < .01), tryptophan 2,3-dioxygenase (p < .05), serpin family F member 1 (p < .01), apolipoprotein A-IV (p < .01) and phosphatidylinositol-glycan-specific phospholipase D (p < .05) were down-regulated in cats with FePAC. Further studies on these potential biomarkers are needed to investigate their diagnostic value.

Neuroprotective effect of carvacrol in an experimental cerebral ischemia and reperfusion rat model

The Neuroprotective effect of carvacrol, which has anti-inflammatory and antioxidant effects, on infarcted cerebral tissue is present in literature, but this contribution was not sufficiently clarified in terms of biochemistry. It is aimed to investigate the effect of orally administered carvacrol on plasma and intraparenchymal levels of TBARS, GSH, SOD, CAT, GSH-Px, IL-1β, IL-4, and TNF-α after the formation of global ischemia in cerebral tissue. Four groups were formed, each containing ten Wistar albino rats. After anesthesia and analgesia, bilateral carotid communis arteries of rats in the first two groups were clamped for 15 minutes with aneurysm clips. Oral 50 mg/kg/day carvacrol was administered for 15 days to the first group (I/R+CRV) of these two groups in which cerebral ischemia-reperfusion (I/R) was established. On the other hand, %0,01 carboxymethylcellulose (CMC), which is a solvent of carvacrol, at a same volume of first group was administered orally for the same duration to the other group in which also I/R was established (I/R+CMC). In the other two groups in which ischemia was not induced, only carotid artery dissections were made and sutured again. In these two groups, 50 mg/kg/day of carvacrol was administered to the first group (CRV). The Same dose of CMC was administered to the second group (control group). After all these treatments, plasma was collected, and brain tissue was dissected from all groups at the end of the 15th day. Carvacrol can be included in the possible treatment regimen of cerebral stroke with the help of other studies that can be supported on this topic.

Neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, and C-reactive protein concentration in female domestic cats subjected to ovariohysterectomy

This study assessed hematological parameters (especially the neutrophil-to-lymphocyte ratio and the platelet-to-lymphocyte ratio) and the C-reactive protein concentration in the plasma of female cats subjected to ovariohysterectomy: prior to surgery as well as 2 and 10 days after surgery. The animals (n = 26) were patients of a private veterinary clinic between July 2019 and March 2021 and were brought there for ovariohysterectomies. The blood was sampled from the cephalic vein into two tubes. One of the tubes was sent to a commercial veterinary laboratory, where a hematological examination was performed. The second tube was centrifuged (2000 g, 10 min). After separation, plasma samples were stored at –80°C. Next, the CRP concentration was determined using an ELISA kit. We noted a statistically significant increase in NLR two days after surgery. The difference in the NLR value before the surgery and after a 10-day recovery period failed to reach statistical significance. Changes in PLR and the CRP concentration during the experiment were insignificant. The results indicate that NLR is an effective marker of stress and inflammation caused by surgery in female cats, whereas the CRP concentration is of no use in this species.