Mastectomy in 25 small ruminants (2002-2019).

OBJECTIVE: To describe the outcome of small ruminants treated with unilateral and bilateral mastectomy by using three surgical techniques. STUDY DESIGN: Retrospective study. ANIMALS: Twenty-five small ruminants (24 goats and one sheep). METHODS: Medical records of animals that underwent mastectomy between November 1, 2002, and May 1, 2019, were reviewed. Follow-up information was obtained by telephone questionnaire with owners. Signalment, surgical data, intraoperative and postoperative complications, bacterial culture results, histopathologic diagnoses, short- and long-term outcomes, and other procedures performed were recorded. RESULT: Procedures consisted of six unilateral (with an elliptical incision) and 19 total (with inverted cloverleaf or elliptical skin incisions) mastectomies. All animals survived to hospital discharge. Intraoperative complications included contamination of the surgical site with mammary-gland fluid, hemorrhage, and difficulty dissecting skin from the mammary gland. Postoperative complications included seroma formation (7/25), surgical-site infection (5/25), and dehiscence of the skin incision (3/25). Mammary neoplasia was diagnosed in seven of 15 animals with histopathologic examination. No association was detected between surgical technique, diagnosis of neoplasia, and long-term outcome. Overall, client satisfaction was high. CONCLUSION: Mastectomy was effective at removing abnormally enlarged udders secondary to chronic mastitis, inappropriate lactation, idiopathic causes, or neoplasia and was associated with a low rate of complications in small ruminants. CLINICAL SIGNIFICANCE: Unilateral mastectomy with an elliptical skin incision or total mastectomy, preferably with inverted cloverleaf skin incision, may be indicated to remove diseased mammary tissue in small ruminants and can result in long-term survival with low morbidity and cosmetically pleasing results.

Determination of blood sirolimus concentrations in liver and kidney transplant recipients using the Innofluor fluorescence polarization immunoassay: comparison with the microparticle enzyme immunoassay and high-performance liquid chromatography-ultraviolet method.

BACKGROUND: Although high-performance liquid chromatography (HPLC) is the method of choice for blood sirolimus determination, the microparticle enzyme immunoassay (MEIA) run on the IMx analyser is widely used in therapeutic monitoring of this immunosuppressant agent. The aim of our study was to evaluate the possible determination of sirolimus using the fluorescence polarization immunoassay (FPIA) commercialized for everolimus quantification. METHODS: Sirolimus concentrations were determined in whole-blood samples from liver and kidney transplant recipients using the Innofluor Certican FPIA (Seradyn Inc.) run on a TDx analyser (Abbott Laboratories), Sirolimus MEIA run on an IMx analyser (Abbott Laboratories), and HPLC (UV detection) methods. RESULTS: The Innofluor FPIA has a similar cross-reactivity with everolimus and sirolimus, and the within- and between-run coefficients of variation obtained for sirolimus determination were 2.7%-13.3%. In analysing different blood samples from liver and kidney transplant patients the linear regressions obtained were: FPIA = 1.12 HPLC + 0.43 (n=104, r=0.874), MEIA = 1.14 HPLC (n=146, r=0.892), and FPIA = 1.00 MEIA + 0.29 (n=106, r=0.941). Better correlation coefficients were obtained between the methods in the liver transplant samples (r>or=0.900) than in the kidney transplant samples (r>or=0.849). No significant effect was found for sirolimus clearance or the blood hematocrit on the relationship between the results produced by both immunoassays and HPLC. CONCLUSION: The Innofluor FPIA is a valid alternative with an analogous performance to the MEIA for the therapeutic monitoring of sirolimus.

Relative proportions of serum carbamazepine and its pharmacologically active 10,11-epoxy derivative: effect of polytherapy and renal insufficiency.

BACKGROUND: The proposed action mechanism and pharmacological activity of carbamazepine (CBZ) and its major metabolite, carbamazepine-10,11-epoxide (CBZE), are the same. The aim of our study was the investigation of the effect of concomitant antiepileptic treatment and renal insufficiency on the relative proportions of serum CBZ and CBZE. METHODS: Serum trough steady-state CBZ and CBZE concentrations were determined by high-performance liquid chromatography (HPLC) in 140 epileptic patients treated with CBZ in monotherapy (n=100) and polytherapy with phenytoin, phenobarbital and valproate (n=40). The levels of CBZ were also determined using the Dade Behring enzyme multiplied immunoassay technique (EMIT). The glomerular filtration rate (GFR) was estimated from serum cystatin C using the Dade Behring nephelometric immunoassay. RESULTS: The CBZE/CBZ and CBZ+CBZE/CBZEMIT ratios were significantly increased in 7 cases (3 in monotherapy and 4 in polytherapy) with GFR<60 mL/min/1.73m2 in relation to the patients treated in monotherapy or polytherapy having normal or mildly decreased renal function (p<0.001). CONCLUSIONS: In patients with moderate to severe renal insufficiency the relative proportion of CBZE with respect to the parent drug is significantly increased. In these cases, the CBZ concentrations obtained using the EMIT, or other immunoassays having low CBZE cross-reactivity, may have an inadequate diagnostic efficiency.

Evaluation of three dosing models for the prediction of steady-state trough clozapine concentrations.

OBJECTIVE: Therapeutic drug monitoring of clozapine may be useful for the clinical management of schizophrenic patients treated with this atypical antipsychotic drug. The aim of our study was the evaluation of three models for the prediction of steady-state trough clozapine concentration. PATIENTS AND METHODS: The trough serum concentrations of clozapine and norclozapine were determined by high-performance liquid chromatography in 296 samples from a group of 21 schizophrenic patients selected for their good therapeutic compliance. Also, the predicted clozapine concentrations were estimated by applying the models of Oyewumi et al. (Ther Drug Monit 1995; 17: 137), Perry et al. (Biol Psychiatry 1998; 44: 733) and Rostami-Hodjegan et al. (J Clin Psychopharmacol 2004; 24: 70). RESULTS: The efficiency for the accurate estimation of clozapine concentrations as subtherapeutic (<240 ng/mL), therapeutic (240-750 ng/mL) or supratherapeutic (>750 ng/mL), using the models of Oyewumi et al., Perry et al., and Rostami-Hodjegan et al., was 82%, 71% and 77% respectively. CONCLUSIONS: The predictive model of Oyewumi et al., which shows an easy calculation way and the greater diagnostic efficiency, may be of clinical value for the prediction of clozapine concentration or the dose required to achieve a specified concentration.

Renal tubular dysfunction in schizophrenic patients treated with antipsychotic drugs.

Several factors have been considered in relation to the free radical formation in schizophrenia, such as the disease itself, drug treatment and smoking. Several chemicals and drugs may cause damage to the renal tubules by different subcellular mechanisms including oxidative stress, and the aim of our study was the investigation of tubular dysfunction in schizophrenic patients. The urinary excretion of beta-N-acetylhexosaminidase (Hex) and its isoenzymes Hex A and Hex B, alpha1-microglobulin, albumin, total proteins and fractionated porphyrins were determined in 45 schizophrenic patients treated with first- and second-generation antipsychotics. In 7 patients, an increase in proteinuria of tubular origin was found, and in one as a result of mixed glomerular/tubular origin. The group of patients had a significantly higher level of excretion than the control group (n = 54) of total Hex (p < 0.001), Hex A (p < 0.05), Hex B (p < 0.001) and the relative proportion of this isoenzyme (p < 0.001). In some cases with normal levels of total Hex and urinary alpha1-microglobulin, the proportion of Hex B was already increased. Significant correlations were found for total Hex and its isoenzymes with alpha1-microglobulin (p < 0.001). Also, the porphyrins had significant correlations with total Hex (p < 0.001), Hex A (p < 0.05), Hex B (p < 0.005) and alpha1-microglobulin (p < 0.001). In the group of patients studied, it was possible to reveal early tubular cell damage (affected structural integrity) with increased excretion of Hex B, possibly mediated by free radicals, previous to the decrease in tubular reabsorption of proteins with low molecular mass filtered by the glomerulus (affected functional integrity).

Comparison of estimated glomerular filtration rates from serum creatinine and cystatin C in patients with impaired creatinine production.

Determination of the glomerular filtration rate (GFR) is important for the drug dosage adjustment and clinical management of patients. The aim of our study was the comparison of estimated GFR values from serum creatinine (eGFRcreatinine) and cystatin C (eGFRcystatin C) in patients with impaired creatinine production. A total of 564 serum samples from patients with kidney disease (n=179), liver (n=71) and kidney (n=182) transplants, critically ill patients (n=82) and healthy subjects (n=50) were analyzed for serum creatinine and cystatin C. The creatinine production rate (CPR) was significantly lower in the different groups of patients than in the control group (p<0.001). A negative correlation was found between the eGFRcreatinine/eGFRcystatin C ratio and CPR (r= -0.964, p<0.001). For CPR higher than 800 mg/24h both procedures for estimating the GFR classified values higher and lower than 60 mL/min with an acceptable agreement; however, for CPR less than 800 mg/24h the eGFRcreatinine led to false negatives in a high number of cases with eGFRcystatin C <60 mL/min.

Serum cystatin C for the prediction of glomerular filtration rate with regard to the dose adjustment of amikacin, gentamicin, tobramycin, and vancomycin.

Dosage regimes of aminoglycosides and vancomycin are modified according to the glomerular filtration rate (GFR). In 130 hospitalized patients who were administered amikacin, gentamicin, tobramycin, and vancomycin by intermittent intravenous infusion, we compared the predicted GFR values from the serum concentrations of creatinine (Cockcroft and Gault. Nephron. 1976;16:31-41) and cystatin C (Larsson et al. Scand J Clin Lab Invest. 2004;64:25-30) with respect to their relevance for proper dosage. In 83% and 67% of the cases, respectively, the serum levels of albumin and cholinesterase were below the corresponding lower limit of the reference range. The ratio of creatinine/cystatin C concentrations presented significant correlations with the predicted rate of creatinine production (r=0.762, P<0.001), serum albumin concentration (r=0.205, P<0.05), and catalytic serum concentrations of cholinesterase (r=0.207, P<0.05), gamma glutamyltransferase (r=-0.273, P<0.01), and alkaline phosphatase (r=-0.289, P<0.01). The GFR (mean+/-SD; median) predicted by the serum creatinine (84.0+/-35.1 mL/min/1.73 m; 82.6 mL/min/1.73 m) was significantly higher (P<0.001) than that predicted by the serum cystatin C (53.1+/-30.2 mL/min/1.73 m; 44.9 mL/min/1.73 m). The ratio between the GFR values predicted by creatinine and cystatin C had a highly significant negative correlation with the rate of creatinine production (r=-0.912, P<0.001). Furthermore, significant differences were found for the peak concentrations and clearances of amikacin and vancomycin estimated by means of the Abbottbase Pharmacokinetic Systems program, and using the GFR values predicted by the serum creatinine and cystatin C (P<0.005). In patients with hepatic dysfunction, the clearance of creatinine predicted by the Cockcroft-Gault formula leads to a significant overestimation of the GFR. Cystatin C seems to be a valid alternative as a GFR marker with regard to drug dose adjustment in these cases.

A comparative study of three kidney biomarker tests in autosomal-dominant polycystic kidney disease.

BACKGROUND: The relationship between the progress of tubular damage and renal insufficiency in autosomal-dominant polycystic kidney disease (ADPKD) is a subject of doubtless interest, and is the object of this present work. METHODS: A total of 92 adult ADPKD patients of both genders were studied, none of which presented end-stage renal disease (ESRD), and classified according to an ultrasound score based on kidney size and number of cysts. Urinary albumin and beta-N-acetylhexosaminidase (Hex) and its isoenzymes were determined, together with serum glutathione peroxidase, cystatin C, creatinine, and urea. RESULTS: A frequent elevation of the urinary Hex was found and an alteration of its isoenzymatic profile, with 31% of the normotensive patients with normoalbuminuria already presenting an increased proportion of Hex B isoenzyme. Keeping age constant, a partial significant correlation was found between the ultrasound score and the proportion of Hex B (r = 0.352, P < 0.05), but not with albuminuria or cystatin C. In 42 patients the different biochemical variables were again determined after 1 year, finding that in the 13 normotensive patients with normoalbuminuria there had been a significant decrease in the concentration of cystatin C (P < 0.05), and a significant increase in the urinary excretion of albumin and Hex B isoenzyme (P < 0.05). By the other hand, in the other 29 patients with micro- or macroalbuminuria and hypertension, no significant differences were found. CONCLUSION: The results point toward an important participation of tubular damage in the pathogenesis of this disease. It may also be suggested that in normotensive and normoalbuminuric ADPKD patients, a gradual increase of glomerular filtration would be produced. After the start of hypertension and microalbuminuria, the glomerular filtration rate (GFR) would decrease progressively, although more slowly.

Clinical significance of hematocrit interference in the tacrolimus II microparticle enzyme immunoassay: a tentative approach.

Recently, the possible interference of hematocrit on the results of the Abbott Tacrolimus II microparticle enzyme immunoassay (MEIA) has been described, although its significance in clinical practice has not been established as yet. The aim of our study was the evaluation of the significance of this analytical interference in therapeutic tacrolimus monitoring. In 1121 cases selected at random over a 9-month period from kidney (n=379) and liver (n=742) transplant patients, an estimation was made of errors caused by the hematocrit in the results provided by the Tacrolimus II MEIA. In accordance with the available data, it was assumed that an error may be produced beyond the range of hematocrit values from 30% to 40%, either positive or negative respectively, of 3% per unit of hematocrit. The acceptance criterion for accuracy was no more than 15% of deviation (error) with respect to the experimental concentration of tacrolimus. In 160 cases (14.3%) the results of the Tacrolimus II MEIA would not be acceptable due to hematocrit-dependent errors, both with positive (hematocrit <25%) in 108 cases (9.7%) and negative values (hematocrit >45%) in 52 cases (4.6%). The obtained results demonstrate the practical interest of the subject, although additional studies are required in order to validate our approach to the clinical significance of this hematocrit-dependent interference in the Tacrolimus MEIA.

More on the reliability of the microparticle enzyme immunoassay (MEIA) for therapeutic tacrolimus monitoring.

Recently, the validity of determining tacrolimus concentrations in blood samples using Abbott's microparticle enzyme immunoassay (MEIA) has been seriously questioned. This communication presents the results obtained in our laboratory using the MEIA for 31 pooled blood samples from kidney and liver transplant patients (Tacrolimus International Proficiency Testing Scheme). A good correlation was found with regard to the average of the MEIA method (r=0.950) without reaching clinical significance for the difference between the means (9.54 ng/ml vs 9.74 ng/ml), although the standard error of the estimate (Syx=0.66 ng/ml) was clinically significant. With regard to the average values for HPLC-methods, a lower correlation coefficient was found (r=0.819), with a difference between the means (9.54 ng/ml vs 8.91 ng/ml) slightly greater than the clinically acceptable value, and a clinically significant standard error of the estimate (Syx=1.22 ng/ml). In the case of the correlation between the average values for the MEIA and HPLC methods, a higher correlation coefficient was found (r=0.918), with a difference between the means (9.74 ng/ml vs 8.91 ng/ml) and a standard error of the estimate (Syx=0.75 ng/ml) being clinically significant. However, these differences were lesser than 15% of HPLC results in accordance with the American Association of Pharmaceutical Scientists acceptance criteria for analytical methods employed for quantification of drugs and their metabolites.

Relationship between serum cystatin C and creatinine in kidney and liver transplant patients.

BACKGROUND: The results obtained for serum cystatin C, which has been proposed as a novel marker of glomerular filtration rate (GFR), in kidney and liver transplant are still very limited. In our study, the relationship between serum cystatin C and creatinine in kidney and liver transplant patients was investigated. METHODS: Serum cystatin C and creatinine concentrations were determined in 182 samples from 52 kidney transplant patients and 71 samples from 28 liver transplant patients at 1-9870 days post-transplantation time. Eighty-seven serum samples from 66 patients with different types of chronic kidney disease were also analysed. RESULTS: The serum creatinine (r=-0.517, p<0.001) and cystatin C (r=-0.409, p<0.001) concentrations were negatively correlated with the post-transplantation time in the kidney transplant patients. In the liver transplant patients, the correlation between these variables is not statistically significant. The creatinine/cystatin C ratio in the liver transplant group is significantly lower than in the other group of patients (p<0.001). This ratio in the kidney transplant patients groups is significantly lower than in the kidney disease group (p<0.001). In the kidney transplant patients the creatinine/cystatin C ratio and the post-transplantation time were negatively correlated (r=-0.523, p<0.001); however, in the liver transplant patients the correlation between these variables was not significant. CONCLUSIONS: In the groups of kidney disease and kidney transplant patients, as renal function decreases, there is an increase in the creatinine/cystatin C ratio. This may be due to the fact that, since creatinine is eliminated by glomerular filtration and tubular secretion, as renal function is impaired, its serum concentration increases to a greater extent than that of cystatin C, which is only eliminated by glomerular filtration. In the liver transplant patients, the creatinine/cystatin C ratio is lower than in the other groups. This may be due to better preserved renal function, lower muscular mass and a reduced rate of creatine formation and creatinine production in some of these patients. The serum cystatin C would be a better GFR marker than the widely used creatinine in liver transplant patients.