Clinical Chemistry Variables Research Articles

Copy Number Variation in the GSTM1 and GSTT1 Genes and the Risk of Liver Cirrhosis in Eastern Ethiopia.

Polymorphisms in glutathione S-transferase M1 (GSTM1) and T1 (GSTT1) can cause an entire gene deletion. The current methodology can accurately identify GSTM1 and GSTT1 copy number variants (CNVs), which may shed light on the true contribution of each gene copy to the cellular detoxification process and disease risk. Because liver cirrhosis is becoming a critical worldwide health issue, this study determined the CNVs of GSTM1 and GSTT1 and their relationship to the risk of liver cirrhosis. In this study, we compared 106 patients with liver cirrhosis to 104 healthycontrols. Real-time PCR was used to identify the CNVs of GSTM1 and GSTT1. Logistic and linear regression models were used to estimate the relationship between liver cirrhosis and clinical chemistry variables with the CNVs, respectively. In 3.3% of the study participants, >2 copies of the GSTM1 or GSTT1 genes were detected. GSTT1 carriers had a significantly lower risk of liver cirrhosis (p<0.05) compared with individuals who had homozygous deletion (adjusted odds ratio (AOR) = 0.47; 95% CI: 0.25, 0.86). This risk reduction was significant (p<0.05) in patients with a single copy of the GSTT1 gene (AOR = 0.48; 95% CI: 0.25, 0.91). Those with ≥2 copies of combined GSTM1 and GSTT1 also had a significantly (p<0.05) lower risk of developing liver cirrhosis compared with double null genotypes (AOR = 0.38; 95% CI: 0.16, 0.91, p trend <0.001). Moreover, ≥2 copies of combined GSTM1 and GSTT1 genes were associated with a substantial decrease in alanine amino transferase (ALT) and aspartate aminotransferase (AST) levels, respectively. A single copy number of GSTT1, and ≥2 copies of combined GSTM1 and GSTT1 genes were associated with a reduced risk of liver cirrhosis in Ethiopians. These findings underscore the importance of gene-environment interactions in the multifactorial development of liver cirrhosis.

A multivariate blood metabolite algorithm stably predicts risk and resilience to major depressive disorder in the general population.

Socioeconomic pressures, sex, and physical health status strongly influence the development of major depressive disorder (MDD) and mask other contributing factors in small cohorts. Resilient individuals overcome adversity without the onset of psychological symptoms, but resilience, as for susceptibility, has a complex and multifaceted molecular basis. The scale and depth of the UK Biobank affords an opportunity to identify resilience biomarkers in rigorously matched, at-risk individuals. Here, we evaluated whether blood metabolites could prospectively classify and indicate a biological basis for susceptibility or resilience to MDD. Using the UK Biobank, we employed random forests, a supervised, interpretable machine learning statistical method to determine the relative importance of sociodemographic, psychosocial, anthropometric, and physiological factors that govern the risk of prospective MDD onset (total n=15,710). We then used propensity scores to rigorously match individuals with a history of MDD (n=491) against a resilient subset of individuals without an MDD diagnosis (retrospectively or during follow-up; n=491) using an array of key social, demographic, and disease-associated drivers of depression risk. 381 blood metabolites and clinical chemistry variables and 4 urine metabolites were integrated to generate a multivariate random forest-based algorithm using 10-fold cross-validation to predict prospective MDD risk and resilience. In unmatched individuals, a first case of MDD, with a median time-to-diagnosis of 72 years, can be predicted using random forest classification probabilities with an area under the receiver operator characteristic curve (ROC AUC) of 0.89. Prospective resilience/susceptibility to MDD was then predicted with a ROC AUC of 0.72 (x˜= 3.2 years follow-up) and 0.68 (x˜= 7.2 years follow-up). Increased pyruvate was identified as a key biomarker of resilience to MDD and was validated retrospectively in the TwinsUK cohort. Blood metabolites prospectively associate with substantially reduced MDD risk. Therapeutic targeting of these metabolites may provide a framework for MDD risk stratification and reduction. New York Academy of Sciences' Interstellar Programme Award; Novo Fonden; Lincoln Kingsgate award; Clarendon Fund; Newton-Abraham studentship (University of Oxford). The funders had no role in the development of the present study.

Aflatoxicosis in Pekin duckling and the effects of treatments with lycopene and silymarin.

Background and Aim:Aflatoxins (AFs) are potent toxic metabolites produced from Aspergillus species. Whose existence in poultry ration leads to drastic economic losses, notably in duck, as the most susceptible poultry species. This study aimed to determine tissue residues of AFs, alterations in selected clinical chemistry variables in serum, mainly during the exposure period, and lycopene and silymarin’s possible roles as herbal treatments against aflatoxicosis in Pekin duckling.Materials and Methods:The study used one hundred and twenty one-day-old Pekin ducklings and classified them into four groups comprising 30 ducklings in each group. The control group (G1) ducklings were fed a mycotoxin-free ration, and G2 received a naturally contaminated ration with 30 ppb of AFs. G3 and G4 consumed contaminated rations with AFs with 30 ppb for 2 weeks and were treated with lycopene 100 mg/kg or silymarin 600 mg/kg/food, respectively, for 10 days. Serum activities of alanine transaminase and alkaline phosphatase (ALP), glutamyl transferase, ALP, total protein and albumin creatinine and uric acid concentrations, oxidant/antioxidant parameters (malondialdehyde [MDA], total antioxidant capacity (TAC), glutathione S-transferase (GST), and catalase [CAT]), and hepatic AFs residue were determined. Lycopene and silymarin were used for the treatment of aflatoxicosis for another 10 days.Results:Hepatic and kidney parameters were elevated in the AFs intoxicated group and reduced in the lycopene- and silymarin-treated groups. They had elevated MDA and AFs residues with decreased antioxidant parameters (TAC, GST, and CAT) in the AFs group. At the same time, treatment with lycopene or silymarin had reversed the action of AFs on MDA, elevated the hepatic residue, and improved antioxidant activity.Conclusion:Lycopene and silymarin, with their potent antioxidant activity, can be used to reverse the harmful effects of AFs on hepatic and kidney tissue.

Comprehensive characterisation of phenolics from Nitraria sibirica leaf extracts by UHPLC-quadrupole-orbitrap- MS and evaluation of their anti-hypertensive activity

Ethnopharmacological relevanceFor more than ten scores years, the leaves and fruits of Nitraria sibirica have been used as a natural remedy for indigestion, irregular manes, and hypertension in the Middle East and Central Asia, especially, are recommended for hypertension treatment in the northwest region, China. Aim of the studywe aimed to support the traditional usage of N. sibirica leaves as pharmaceuticals or dietary supplements in treatment of hypertension by investigating their chemical constituents and anti-hypertensive activity. MethodsWe identified the chemical composition of N. sibirica leaves ethanolic purified extract (NSL-EPE) using UHPLC-quadrupole-orbitrap-MS, and quantified the main chemical constituents by an analytical method established and validated. We also evaluated anti-hypertensive activity of NSL-EPE using spontaneously hypertensive rats (SHR): blood pressure was measured weekly by non-invasive blood pressure (NIBP) measurements; hemodynamic parameters, biochemical and clinical chemistry variables in plasma, serum and kidney tissue were measured after 10 weeks of treatment with NSL-EPE as well. ResultsUHPLC-quadrupole-orbitrap-MS analysis identified 52 compounds, of which 40 compounds were reported for the first time in N. sibirica. 11 phenolic compounds further quantitatively analyzed, among which the most abundant compound was found to be clovin (8.8%). Systolic blood pressure decreased progressively from the second treatment week compared to that in non-treated SHRs. The plasma endothelin, aldosterone, angiotensin II levels were significantly increased, while the level of NOX was significantly decreased; glutathione to oxidized glutathione ratio, superoxide dismutase and total catalase levels in the kidney tissue were markedly accelerated, while malondialdehyde level was significantly reduced in NSL-EPE treated SHRs. Moreover, the serum cholesterol, triglyceride, blood uria nitrogen and creatinine were attenuated in NSL-EPE treated SHRs (P < 0.05), but in sharp contrast to those values in the water-treated SHRs. ConclusionThis study screened out leading compounds from N. sibirica and offered a new understanding of the antihypertensive properties of N. sibirica leaves, by which inhibit oxidative stress-induced endothelial dysfunction and improve lipid profiles.

The Influence of a Health-Related Fitness Training Program on Motor Performance as Well as Hematological and Biochemical Parameters.

The study was aimed at designing a health exercise program appealing to inactive young men, and then testing the men’s metabolic responses to the program using common diagnostic markers of general health. Six men, aged 22–29 years, took a part in training program to increase their motor performance and improve general health conditions. Body composition parameters, clinical chemistry variables (metabolites, albumin, total protein, ferritin, C reactive protein, lipid profile, ions, and selected enzymes activities) and blood morphology parameters were determined. Motor performance measured before and after a 4-month-long macrocycle indicated an increase in endurance, pace, and agility of the participants. Significant differences were found in analyzed enzymes activities. There was a significant increase in C-reactive protein levels from pre- to post-training. Additionally, changes in hematological biomarkers were seen that suggest erythropoiesis might significantly increase, specifically during the last 2-month-long mesocycles. The proposed training program induced small improvements in endurance, pace, and agility. It was also confirmed that changes in aspartate (AST) and alanine (ALT) activities emerge before any increase in creatine kinase (CK) activity that is important in monitoring of the training loads. Observed changes in red blood cell-related parameters suggest increase in erythropoiesis in the second half of the training cycle.

Evaluating the Relationship Between Lean Liver Volume and Fat-Free Mass.

Fat-free mass has gained wide acceptance as a scaler of the maintenance dose rate in obese patients. The choice of fat-free mass as a size scaler for the maintenance dose rate is based on its relationship with drug clearance, on the basis that only lean tissue is sufficiently metabolically active to provide capacity for elimination. For xenobiotics, the majority of biotransformation occurs in the liver and hence fat-free mass is implied to scale linearly with the component of liver that is metabolically active. The liver, like the body, can be assumed to comprise two components, lean mass and fat mass. We expect the lean liver mass (or volume) to be the component that most closely relates to drug clearance. The objective of this study was to investigate the relationship of lean liver volume and fat-free mass. Total liver volume and liver fat volume were measured in 100 Indian adults by computed tomography. Lean liver volume was derived as the difference between the two measurements (as liver volume - liver fat volume). Covariate modelling to describe lean liver volume, using NONMEM version 7.3, involved testing the influence of body weight, sex, body surface area and fat-free mass with or without allometric scaling (by estimating the exponent) and the influence of clinical chemistry variables. The final model did not exclude a linear relationship between lean liver volume and fat-free mass, while allometric scaling by body weight0.75 was also supported by the data. While scaling by fat-free mass, the coefficient of proportionality (i.e. lean liver volume per kg fat-free mass) was higher in female (31.25mL) than male (25.81mL) subjects. A model to predict lean liver volume from readily available patient data was developed and evaluated. Fat-free mass plus sex was found to be the best body descriptor to scale lean liver volume. The utility of this model in scaling drug clearance and dose requirements of hepatically cleared drugs needs further exploration.

Human mercaptalbumin and nuclear factor kappa B in subjects with mildly impaired renal, vascular and metabolic health compared to “super-healthy” subjects: the BIOCLAIMS Integration study

The redox state of human albumin is known to be affected by several diseases, while data is limited on activation of NF-kB, and absent in mildly impaired human health. The aim of this study was to compare 4 contrasting groups of the BIOCLAIMS cohort: mildly impaired renal health (group 1, estimated glomerular filtration rate, MDRD, 30-60 ml/min/1.73 m2, n=43), vascular health (group 2, intima-media thickness, IMT, left and right >75th percentile, n=59), metabolic health (group 3, HOMA index >2.5 and HbA1c 38.8 -44 mmol/mol, n=44), and “super” healthy subjects (group 4, all clinical chemistry variables within normal range 10%, IMT not >75th percentile at both sides, not on medications, n=56). Plasma human mercapt-(HMA) and non-mercaptalbumin (HNA-1) were determined by HPLC; activation of NF-kB in PBMC by TransAM Active Motif assay. ANCOVA, with age (group 1>2,4; 3>4) as covariate, showed that HMA and HMA:HNA-1 ratios were in group 1 3 (P4 (P

Eight weeks of resistance training in conjunction with glutathione and L-Citrulline supplementation increases lean mass and has no adverse effects on blood clinical safety markers in resistance-trained males

BackgroundSupplementation of combined glutathione (GSH) with L-citrulline in response to a single bout of resistance exercise has been shown to increase plasma nitric oxide metabolites, nitrite and nitrate and cyclic guanosine monophosphate (cGMP), which may play a role in muscle protein synthesis. As a result, in response to resistance training (RT) these responses may establish a role for GSH + L-citrulline to increase muscle mass.This study attempted to determine the effects of an 8-week RT program in conjunction with GSH (Setria®) + L-citrulline, L-citrulline-malate, or placebo supplementation on lean mass and its association with muscle strength. The secondary purpose was to assess the safety of such supplementation protocol by assessing clinical chemistry markers.MethodsIn a randomized, double-blind, placebo-controlled design, 75 resistance-trained males were randomly assigned to ingest GSH + L-citrulline (GSH + CIT), L-citrulline-malate, or cellulose placebo daily while also participating in 8 weeks of RT. The full dose of each supplement was delivered in capsules that were identical in weight, size, shape, and color. Participants completed testing sessions for body composition and muscle strength before and after 4 and 8 weeks of RT and supplementation. Venous blood samples were obtained before and after 8 weeks.ResultsLeg press was increased with RT but was not significantly different between groups (p > 0.05); however, bench press strength was not increased with RT (p > 0.05). There were no significant changes in total body mass, fat mass, or total body water during 8 weeks of RT and supplementation. Lean mass increased in both GSH + CIT when compared to PLC; however, the increase was significant only after 4 weeks. Lean mass and strength were positively correlated (p < 0.05) in GSH + CIT, but not CIT-malate or PLC. Neither RT nor supplementation had any significant effects on blood clinical chemistry variables (p > 0.05).ConclusionCompared to PLC, supplementation of GSH + CIT during resistance training increased lean mass after 4 weeks of RT and was positively associated with muscle strength. However, after 8 weeks of RT there were no significant differences in any of the measured variables.

Evaluation of a fixed-dose combination of benazepril and pimobendan in dogs with congestive heart failure: a randomized non-inferiority clinical trial.

A fixed-dose combination tablet of benazepril and pimobendan (Fortekor Plus; Elanco Animal Health) was tested in dogs with congestive heart failure (CHF) caused by myxomatous mitral valve disease (MMVD) in a three-arm, masked, randomized, non-inferiority clinical trial in Japan. The test group (n = 34) received Fortekor Plus twice daily. Two control groups received registered formulations of benazepril (Fortekor; Elanco Animal Health) and pimobendan (Vetmedin; Boehringer Ingelheim Vetmedica) with administration of Vetmedin twice daily and Fortekor twice (Control I, n = 14) or once (Control II, n = 19) daily. Diuretics were used in 22 dogs (32.8%). Global clinical scores decreased significantly from baseline in all groups; there were no significant differences between groups, and non-inferiority of Fortekor Plus compared to Control I, Control II, and combined Control I + II groups was demonstrated. There were no significant differences between groups for relevant clinical chemistry and hematology variables or frequency of all adverse events. Frequency of emesis was significantly (p = 0.0042) lower in the Fortekor Plus (8.8%) group than in the Control I + II (39.4%) group. In conclusion, Fortekor Plus had non-inferior efficacy and was associated with significantly less emesis compared to Fortekor and Vetmedin in dogs with CHF caused by MMVD.

The Impact of the Progressive Efficiency Test on a Rowing Ergometer on White Blood Cells Distribution and Clinical Chemistry Changes in Paralympic Rowers During the Preparatory Stage Before the Paralympic Games in Rio, 2016 - A Case Report.

There is a large gap in knowledge regarding research on post-exercise blood changes in disabled athletes. There are relatively few data on adaptive mechanisms to exercise in disabled athletes, including disabled rowers. Two rowers from a Polish adaptive rowing settle TAMix2x that qualified for the Paralympic Games in Rio, 2016 took part in this study. They performed a progressive test on a rowing ergometer until exhaustion. The cardiorespiratory fitness measures, complete blood count, white blood cells’ distribution and 30 clinical chemistry variables describing laboratory diagnostic profiles and general health were determined. The extreme effort induced changes in all studied metabolites (glucose, creatinine, urea, uric acid, total and direct bilirubin), albumin, total protein levels in both participants. Furthermore, a post-exercise increase in aspartate transaminase activity, yet a 2-fold decrease during the recovery time in both rowers were found. White blood cell count increased 2-fold after the test. The percentages of natural killer cells were higher and total T lymphocytes were lower after the exercise protocol. There were higher percentages of suppressor/cytotoxic and lower percentages of helper/inducer T lymphocyte subsets in both studied rowers. No changes in B lymphocytes distribution were observed. Lack of inflammatory symptoms during the experiment suggests a high level of rowers’ biological adaptation to the physical effort. The different changes in physiological, biochemical and immunological variables are related to the adaptive mechanism to physical exercise allowing for improvement of performance.

Evaluation of cardiovascular effects of intravenous robenacoxib in dogs.

The objective of the study was to assess the cardiovascular effects of intravenous (IV) dosing with robenacoxib (Onsior® ) in conscious adult healthy beagle dogs. The study employed a randomized, open, placebo-controlled, four-phase Latin square design. A total of eight dogs received a single dose of 2mg/kg and 4mg/kg IV robenacoxib (test groups), 2mg/kg subcutaneous (SC) robenacoxib (reference dose and route), and IV isotonic saline (control). There were no significant differences between groups for clinical observations, buccal mucosal bleeding time or blood hematology, coagulation, and clinical chemistry variables in all eight dogs. In a subset of four dogs, no significant differences between groups were detected using telemetric assessment for arterial blood pressure, heart rate, electrocardiogram, or body temperature over 8hr postdose. In conclusion, no significant cardiovascular effects were detected after a single IV dose of 2 or 4mg/kg robenacoxib in conscious healthy dogs.

Toxicological characterisation of two novel selective aryl hydrocarbon receptor modulators in Sprague-Dawley rats

The aryl hydrocarbon receptor (AHR) mediates the toxicity of dioxins, but also plays important physiological roles. Selective AHR modulators, which elicit some effects imparted by this receptor without causing the marked toxicity of dioxins, are presently under intense scrutiny. Two novel such compounds are IMA-08401 (N-acetyl-N-phenyl-4-acetoxy-5-chloro-1,2-dihydro-1-methyl-2-oxo-quinoline-3-carboxamide) and IMA-07101 (N-acetyl-N-(4-trifluoromethylphenyl)-4-acetoxy-1,2-dihydro-5-methoxy-1-methyl-2-oxo-quinoline-3-carboxamide). They represent, as diacetyl prodrugs, AHR-active metabolites of the drug compounds laquinimod and tasquinimod, respectively, which are intended for the treatment of autoimmune diseases and cancer. Here, we toxicologically assessed the novel compounds in Sprague-Dawley rats, after a single dose (8.75–92.5mg/kg) and 5-day repeated dosing at the highest doses achievable (IMA-08401: 100mg/kg/day; and IMA-07101: 75mg/kg/day). There were no overt clinical signs of toxicity, but body weight gain was marginally retarded, and the treatments induced minimal hepatic extramedullary haematopoiesis. Further, both the absolute and relative weights of the thymus were significantly decreased. Cyp1a1 gene expression was substantially increased in all tissues examined. The hepatic induction profile of other AHR battery genes was distinct from that caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The only marked alterations in serum clinical chemistry variables were a reduction in triglycerides and an increase in 3-hydroxybutyrate. Liver and kidney retinol and retinyl palmitate concentrations were affected largely in the same manner as reported for TCDD. In vitro, the novel compounds activated CYP1A1 effectively in H4IIE cells. Altogether, these novel compounds appear to act as potent activators of the AHR, but lack some major characteristic toxicities of dioxins. They therefore represent promising new selective AHR modulators.

Evaluation of injectable robenacoxib for the treatment of post-operative pain in cats: results of a randomized, masked, placebo-controlled clinical trial.

BackgroundFew pharmaceuticals are registered in cats for the management of post-operative pain and inflammation. The objective of this study was to assess the field efficacy and safety of an injectable formulation of the nonsteroidal anti-inflammatory drug robenacoxib in cats undergoing surgery. The study was a multi-center, prospective, randomized, masked, parallel-group, placebo-controlled clinical trial. A total of 349 cats were enrolled and underwent surgery comprising forelimb onychectomy, as an example of orthopedic surgery, plus either ovariohysterectomy or castration. All cats received butorphanol prior to anesthesia and forelimb four-point regional nerve blocks with bupivacaine after induction of general anesthesia. Cats were randomized to receive daily subcutaneous (s.c.) injection of robenacoxib, at a target dosage of 2.0 mg/kg (n = 174), or placebo (n = 175) once prior to surgery and for an additional two days post-operatively.ResultsSignificantly (P = 0.037) fewer cats administered robenacoxib received additional analgesia rescue therapy (34 of 173, 19.7 %) compared to cats given placebo (73 of 175, 41.7 %). The percentage of treatment success was therefore 80.3 % with robenacoxib and 58.3 % with placebo. Behavior, posture, pain on palpation of the paw and soft tissue surgery sites, and overall pain were significantly (P < 0.05) improved versus placebo at various time points within the first 8 h in cats receiving robenacoxib. The most frequently reported adverse events were incision site infection/dehiscence, bleeding, vomiting, decreased appetite and lethargy. Frequencies of reported adverse clinical signs, hematology, serum chemistry and urinalysis variables, and body weight changes were similar between groups. There were no significant changes from baseline with robenacoxib in hepatic, hematological or renal clinical pathology variables.ConclusionsRobenacoxib by s.c. injection was effective and well tolerated in the control of post-operative pain associated with orthopedic, ovariohysterectomy and castration surgery in cats.

Hematologic and serum biochemical changes associated with Devil Facial Tumor Disease in Tasmanian Devils.

Devil Facial Tumor Disease (DFTD) is an infectious tumor causing significant population declines in wild Tasmanian Devils. While clinical assessment and pathology have been well reported for DFTD, there is a lack of information on hematologic and biochemical alterations associated with DFTD. The purpose of the study was to determine hematologic and serum biochemical variation in healthy, wounded, and DFTD-affected Tasmanian Devils. Blood samples were collected from wild Tasmanian Devils at 5 sites in Tasmania. Hematology and clinical biochemistry variables were compared between clinically healthy, wounded, and DFTD-affected devils. Differences were also analyzed among stages of DFTD, including individuals pre- and postclinical signs developing, and between ulcerated and nonulcerated DFTD tumors. Statistically significantly increased counts in WBC, neutrophils, and platelets, and concentration of fibrinogen, as well as decreased counts in lymphocytes, erythrocytes, and HGB concentration were observed in DFTD-affected devils compared to healthy devils. Activities of ALP, ALT, and GLDH, concentrations of sodium, potassium and albumin, and sodium-to-potassium ratio and albumin-to-globulin ratio were significantly lower, and AST activity was significantly higher in animals with DFTD when compared to clinically healthy animals. No significant differences were found among stages of DFTD or ulcerated and nonulcerated tumors. The differences in hematology and clinical chemistry variables in devils with DFTD compared to healthy devils are nonspecific and reflective of acute phase response and inflammation, and anemia of chronic disease. Similar changes are observed with wounds but to a lesser extent.

Effect of benazepril, robenacoxib and their combination on glomerular filtration rate in cats.

BackgroundCombined use of angiotensin-converting enzyme inhibitors and nonsteroidal anti-inflammatory drugs may induce acute kidney injury in humans, especially when combined with diuretics. The objective of this investigation was to evaluate the effects of benazepril, robenacoxib and their combination in healthy cats.In each of two studies (study 1 followed by study 2), 32 healthy cats were randomised to one of four groups (n = 4 male and 4 female cats per group) in a parallel-group design. The groups received orally once daily for 7 days either placebo (control group), benazepril, robenacoxib or benazepril plus robenacoxib. In study 2, all groups received in addition 0.5 mg/kg furosemide twice daily by subcutaneous injection for 7 days.ResultsBenazepril, robenacoxib and their combination were well tolerated as evidenced from lack of clinical signs and no negative effects on body weight, feed consumption and clinical chemistry, haematology and urinalysis variables.The primary endpoint of the study was the glomerular filtration rate (GFR), which was estimated from the plasma clearance of iohexol. In the absence of furosemide, GFR was significantly higher in cats receiving the combination of benazepril plus robenacoxib compared to the other three groups, and was also significantly higher in females receiving only benazepril compared to the control.Administration of furosemide induced diuresis, reduced GFR and activated the renin-aldosterone-angiotensin system, evidenced from increased plasma renin activity and plasma aldosterone concentrations. Compared to the control group in cats treated with furosemide, GFR was increased by benazepril (females only) but decreased by robenacoxib (males only). Benazepril, robenacoxib and their combination significantly inhibited the increase in plasma aldosterone induced by furosemide.ConclusionsThe combination of benazepril and robenacoxib was well tolerated and either increased or had a neutral effect on GFR in healthy cats without or with concomitant furosemide. The combination of benazepril and robenacoxib reduced plasma aldosterone concentrations increased by furosemide. It is recommended to test the efficacy and safety of the combined use of benazepril and robenacoxib in cats with clinical disease, notably proteinuric chronic kidney disease.

Determination of liver and muscle aflatoxin B1 residues and select serum chemistry variables during chronic aflatoxicosis in broiler chickens.

Aflatoxins are secondary metabolites of the fungus Aspergillus sp. The presence of aflatoxin in poultry feeds results in direct toxicity and economic losses, and human health hazards after consumption of contaminated liver and meat. The study was conducted to assess tissue residues of aflatoxin B1 (AFB1), and alterations in select clinical chemistry variables in serum during chronic aflatoxicosis in broiler chicks fed different dietary levels of AFB1. Six groups of broiler chickens were fed diets containing between 0 and 800 ppb of AFB1 for 28 days. Groups of birds were terminated on days 0, 5, 13, 15, 20, and 28, and AFB1 levels were determined by HPLC in liver and muscle. Serum activities of ALT and ALP, and total protein and albumin concentrations were determined. No AFB1 residues were detected in liver after 50 ppb AFB1, and muscle after 50 and 100 ppb AFB1 feeding. Residues above the permissible threshold (> 2.0 ng/g) were only detected in liver tissues of groups fed 400 ppb and 800 ppb AFB1 in feed. The ALT and ALP activities in treated groups were significantly higher, and total protein and albumin concentrations were significantly lower in all treated groups compared to controls. Continuous feeding of AFB1 to broiler chicken at levels of 50 and 100 ppb for 28 days did not reveal measurable AFB1 residues in muscle tissues. Serum values of ALT, ALP, total protein, and albumin may serve as markers for chronic aflatoxicosis in affected poultry.

The effects of a ration change from a total mixed ration to pasture on health and production of dairy cows

In pasture-based dairy production systems, dairy cows often receive a silage- and concentrate-based ration [total mixed ration (TMR)] during wintertime and are gradually introduced to fresh herbage in spring. The present study aimed to investigate how the transition to this new nutritional situation influenced different production and health indicators. A 10-wk trial was performed in spring 2014, including 60 dairy cows of the German Holstein breed (166±23d in milk, 23.5±3.7kg of milk/d; means ± SD). The cows were divided into a pasture and a confinement group (PG and CG, respectively). The CG stayed on a TMR-based diet (35% corn silage, 35% grass silage, 30% concentrate; DM basis), whereas the PG was gradually transitioned from a TMR- to a pasture-based ration (wk 1=TMR-only, wk 2=3 h/d on pasture, wk 3 and 4=12 h/d on pasture, wk 5–10=pasture-only). A continuous grazing system was implemented on a ryegrass dominated pasture and temperature humidity indices were assessed based on continuous recording of temperature and humidity indoors as well as outdoors. Dry matter intake (DMI) from TMR, milk production, body weight (BW), and body condition score decreased as soon as the PG had partial access to pasture. Milk production and BW decreased even further in the first week on a full grazing ration, but thereafter BW increased again and milk production stabilized. The DMI estimation using the n-alkane method in wk 7 and 9 revealed an increase in DMI from pasture between the 2 time points and indicates an adaptation of grazing behavior and metabolism over several weeks. Increased serum β-hydroxybutyrate and fatty acids concentrations at several time points, as well as a continuous body condition score decrease during the whole course of the trial, indicate an energy deficit in the PG. A significant correlation between serum glucose concentrations and the temperature humidity indices was observed. An increase in serum and milk urea concentrations as well as an increase in the urine total N to creatinine ratio occurred in the PG. To assess possible negative effects of the ration change on metabolic and liver health, different clinical chemistry variables and complete blood counts were assessed. No biologically relevant changes were observed for serum albumin, total protein, cholesterol, aspartate transaminase, γ-glutamyltransferase, and glutamate dehydrogenase concentrations, as well as for white and red blood cell counts.

Clinical safety of robenacoxib in feline osteoarthritis: results of a randomized, blinded, placebo-controlled clinical trial.

The objective of this study was to evaluate the clinical safety of the non-steroidal anti-inflammatory drug (NSAID) robenacoxib in cats with osteoarthritis. Degenerative joint disease, including osteoarthritis, is highly prevalent in cats and many cases have associated pain and impaired mobility. Although NSAIDs are used routinely to control pain and inflammation in cats with osteoarthritis, there are safety concerns because of the high concurrent prevalence of chronic kidney disease (CKD) and the paucity of data on the safety of these drugs in target clinical populations. A total of 194 cats with osteoarthritis were recruited and randomly allocated to receive either robenacoxib at a dosage of 1.0-2.4 mg/kg (n = 95) or placebo (n = 99) tablets PO q24h for 28 days. Safety was assessed in 193 cats, including a subgroup of 40 animals with concurrent CKD, defined as serum creatinine concentration ⩾1.6 mg/dl and urine specific gravity <1.030. Safety endpoints included reports of adverse events, results of clinical examinations, including body weight, and clinical chemistry and hematology variables. In all 193 cats and the subgroup of 40 animals with concurrent CKD, there were no differences between groups in frequencies of reported adverse events, body weight change or results of serum or urine chemistry or hematology variables. Robenacoxib was well tolerated when administered daily for 1 month in cats with osteoarthritis, including cats with evidence of concurrent CKD. There was no clinical indication of damage to the gastrointestinal tract, kidney or liver.

Nutritional effects of egg shell membrane supplements on chicken performance and immunity

Eggshell membranes (ESM) contain a variety of proteins and peptides which help in the development of embryo and provide protection to it. Many of the peptides and proteins associated with ESM have antimicrobial, immune-modulatory, and adjuvant properties. We hypothesized that the membrane byproducts from egg, provided as posthatch nutritional supplements to chickens, may improve their performance and immunity. To explore its effect, we fed 3 groups of broiler chicks with feed containing 0, 0.2, and 0.4% ESM from d 1 posthatch through 14 d and regular feed thereafter. The birds were individually weighed at the onset of the study and at weekly intervals until the termination at third wk when they were bled and euthanized. The relative weights of liver, spleen, bursa, and heart, hematology profiles, and clinical chemistry variables including serum IgM, IgG, and corticosterone concentrations were measured. The chickens in the ESM treated groups showed a statistically significant increase in BW with no impact on relative organ weights. Compared with controls, the WBC and lymphocyte percentage increased in chickens fed 0.4% ESM whereas the monocyte percentage decreased at both levels of ESM. Except for the serum protein which increased in ESM fed birds no other metabolic clinical chemistry variables showed any significant change. Both IgM and IgG(Y) levels were elevated and corticosterone levels reduced in chickens fed ESM supplemented diets. Our results suggest that ESM supplements during the early phases of growth may improve immunity and stress variables, and enhance their growth performance without any detrimental effect on other physiological parameters.

Robenacoxib versus meloxicam for the control of peri-operative pain and inflammation associated with orthopaedic surgery in cats: a randomised clinical trial

BackgroundNon-steroidal anti-inflammatory drugs (NSAIDs) are widely used in veterinary medicine. Robenacoxib is a NSAID with high selectivity for the cyclo-oxygenase-2 enzyme. In this study, the efficacy and safety of robenacoxib were evaluated in a prospective, randomised, active- and placebo-controlled masked clinical trial in 147 cats undergoing orthopaedic surgery. Cats were randomised into two treatment groups: Group 1, robenacoxib (2 mg/kg) administered via subcutaneous (s.c.) injection before surgery, followed by robenacoxib tablets (1–2.4 mg/kg) administered post-operatively for approximately 9 days (n = 101) and Group 2, meloxicam (0.3 mg/kg) administered s.c. before surgery, followed by placebo tablets administered post-operatively for approximately 9 days (n = 46). Cats were assessed using numerical rating scales (NRSs) by clinicians before surgery and at 3, 8, 22 and 28 hours after surgery and at the final visit (VF on approximately Day 10), and daily by their owners from Day 1 to the VF.ResultsThe primary end point was the global investigator score which was the sum of clinician NRSs for posture, behaviour and pain on palpation/manipulation. The efficacy of the single robenacoxib injection, assessed during 3 to 22 hours, was statistically non-inferior to meloxicam, with a relative efficacy of 1.029 (95% confidence interval, 0.847–1.231). No significant differences were detected during the follow-up treatment with robenacoxib tablets for approximately 9 days compared with placebo via clinician assessments at 28 hours and the VF, or in owner assessments on Days 1–VF. There were no significant differences in frequencies of reported adverse events, clinical observations and haematology or clinical chemistry variables between the groups.ConclusionsSingle s.c. injection of robenacoxib before surgery had non-inferior efficacy compared with meloxicam in controlling post-operative pain and inflammation in cats undergoing orthopaedic surgery. Follow-up treatment with oral robenacoxib tablets for approximately 9 days was well tolerated, but there were no differences in the efficacy scores after Day 1 compared with the group receiving meloxicam s.c. followed by placebo control.