Comparison of free thyroxine measurement by chemiluminescence and equilibrium dialysis following 131I therapy in hyperthyroid cats.

OBJECTIVES: The first objective was to assess correlation between free thyroxine (fT4) measurements by equilibrium dialysis (fT4ED; Antech Diagnostics) and a chemiluminescent enzyme immunoassay (fT4CEIA; IMMULITE 2000 Veterinary Free T4 [Siemens Healthcare Diagnostics Products]) in hyperthyroid, otherwise healthy, cats before (T0), and 1 month (T1) and 11-23 months (T2) after radioactive iodine (131I) therapy. The second objective was to determine correlation between thyroid status based on fT4 (by both techniques) and the gold standard, thyroid scintigraphy. METHODS: Thyroid status, including thyroid-stimulating hormone (TSH), total thyroxine (TT4) and fT4 serum concentrations, were assessed in 45 client-owned hyperthyroid cats before (T0), and 1 month (T1) and 11-23 months (T2) after 131I therapy. fT4 was determined by a chemiluminescent enzyme immunoassay (CEIA) and equilibrium dialysis (ED). Quantitative thyroid scintigraphy (with sodium 99m-Tc-pertechnetate) was performed at T2. RESULTS: Spearman correlation between fT4CEIA and fT4ED was 0.81, 0.88 and 0.79 at T0, T1 and T2, respectively. fT4CEIA was consistently lower than fT4ED, with a median difference of -5.4 pmol/l (P <0.001) and -4.9 pmol/l (P <0.0001) at T1 and T2, respectively. At T2, all cats were identified as euthyroid based on thyroid scintigraphy. None of the cats were identified as being hypothyroid, based on serum TT4 and TSH measurements. Nine of 22 (40.9%) cats had an fT4CEIA below the reference interval (RI) at T2, whereas only 2/22 (9.1%) cats had an fT4ED concentration below the RI at T2. CONCLUSIONS AND RELEVANCE: Good correlation exists between both assays at T1 and T2, but a significant systematic difference is noted at both time points. This could be an indication for reconsideration of the current RI, although further studies are warranted for assessing test accuracy (in otherwise healthy cats and cats with non-thyroidal illness). At this time, routine use of fT4CEIA after 131I therapy is not advised in feline patients.

Effect of dialysis of dog semen on sperm characteristics and some biochemical components of seminal plasma.

The aim of this study was to investigate the effect of dog semen dialysis on sperm characteristics and some biochemical components of seminal plasma. Whole ejaculates were dialyzed against Tris-citrate-fructose extender for a 5 h period at room temperature (using semi-permeable cellulose tubing of 12-14 kDa molecular weight cut-off). It has been demonstrated that the long-term dialysis of dog semen causes a significant decrease in sperm quality parameters and disrupts the biochemical properties of seminal plasma. This procedure requires further improvement.

Effect of dialysis on the proacrosin/acrosin system and motility of turkey (Meleagris gallopavo) spermatozoa during liquid storage.

1. The effect of dialysis on the proacrosin/acrosin system and motility of turkey spermatozoa were examined after 24 and 48 h of liquid storage at 4°C. 2. Fifteen pools of semen diluted in extender were dialysed against Clemson Turkey Semen Diluent (dialysed semen) or stored in aerobic conditions (undialysed semen). Semen quality was assessed by measuring spermatozoa motility, amidase activity of spermatozoa suspension, spermatozoa extract and seminal plasma and anti-trypsin activity of seminal plasma. 3. Extracted amidase activity of dialysed semen was lower than undialysed by 28%. Higher values for speed parameters of spermatozoa were found in dialysed semen in comparison to undialysed, for example, 81.6 µm/s versus 75.0 µm/s for straight-line velocity (VSL), 114.7 µm/s versus 110.3 µm/s for curvilinear velocity (VCL) and 86.6 µm/s versus 79.8 µm/s for average path velocity (VAP). 4. It was concluded that dialysis caused lower amidase activity of spermatozoa and increased speed parameters of progressively motile turkey spermatozoa during storage. Lower extracted amidase activity of dialysed semen reflected better membrane integrity of dialysed semen and suggests that the proacrosin/acrosin system of dialysed spermatozoa is less susceptible to activation compared to undialysed semen.

Feline acute kidney injury: 1. Pathophysiology, etiology and etiology-specific management considerations.

PRACTICAL RELEVANCE: Acute kidney injury (AKI) is a frequently recognized disease process in cats that requires immediate and aggressive intervention. A thorough understanding of the pathophysiologic processes underlying AKI and familiarity with the most common etiologies are essential for providing the most effective and timely therapy. Possessing this knowledge will also allow a more accurate prognosis to be given, and afford the best chance of a favorable outcome. CLINICAL CHALLENGES: Feline patients often present with vague signs of AKI, which may delay treatment and adversely affect the prognosis. Their response to injury and treatment is often different to that of other species. AUDIENCE: This two-part review article is directed at small animal practitioners as well as specialists. Part 1 reviews mechanisms underlying AKI in the cat, as well as etiologies and treatments related to some specific causes of AKI. EVIDENCE BASE: The veterinary literature is limited with regards to the pathophysiology of AKI unique to the cat. However, there are numerous feline studies evaluating causes of AKI.

Feline acute kidney injury: 2. Approach to diagnosis, treatment and prognosis.

PRACTICAL RELEVANCE: Feline acute kidney injury (AKI) is a commonly recognized problem in small animal practice that requires prompt diagnosis and directed therapy. There are many treatment methods with which practitioners should be familiar, including medical options, surgical interventions and renal replacement therapy (dialysis). It is important to know which option is most appropriate for each cause and stage of AKI to deliver the most effective therapy. CLINICAL CHALLENGES: AKI can cause vague clinical signs, but a vast array of life-threatening sequelae. Rapid recognition of potential complications and knowledge of treatment options is imperative for successful management. Feline patients also require an understanding of their unique physiology as it relates to the therapeutic plan. AUDIENCE: This two-part review article is directed at small animal practitioners as well as specialists. Part 2 discusses the diagnosis of AKI in cats using physical examination findings, clinicopathologic results and imaging modalities. The treatment of AKI and its sequelae is also reviewed, with information on recent advances in this area. EVIDENCE BASE: While there is very limited data comparing the outcomes of various treatment options, there is literature addressing the use of several medications, as well as renal replacement therapy, in cats.

Factors influencing the performance of an interferon-γ assay for the diagnosis of tuberculosis in goats.

The interferon-gamma (IFN-γ) assay is an effective tool for the diagnosis of tuberculosis (Tb) in goats. The objectives of this study were to evaluate factors that might affect assay performance: (1) the phenol concentration of the purified protein derivative (PPD, tuberculin) used; (2) dialysis of PPD; and (3) delaying antigenic stimulation of blood samples for 8, 16 and 24h after collection. The assay was performed in duplicate with two cut-off points. Dialysis of PPD reduced test sensitivity, whereas the concentration of phenol did not significantly affect test outcome. Delaying antigenic stimulation of samples >8h resulted in a reduction in test sensitivity, compromising the capacity of the assay to detect infected animals. Performing the assay in duplicate was unnecessary, which has implications for reducing assay costs. These findings will facilitate the effective application of the IFN-γ assay as an ancillary test in Tb eradication programmes in goats.

Adequação de um sistema de hemodiálise em eqüinos / Adequacy of hemodialysis system in horses

Realizou-se a adequação da técnica de hemodiálise com eqüinos, distribuídos em quatro grupos experimentais de seis animais cada. Os animais do grupo I foram submetidos a cateterismo central unilateral (grupo-controle); os do grupo II foram submetidos a cateterismo central unilateral com cateter duplo-lúmen e a uma sessão de hemodiálise de seis horas; os do grupo III a cateterismo central unilateral com cateter duplo-lúmen e duas sessões de hemodiálise de seis horas e os do grupo IV a cateterismo central bilateral com cateter monolúmen e a uma sessão de hemodiálise de seis horas. Empregaram-se xilazina 10 por cento (0,4mg/kg) e acepromazina 2 por cento (0,008mg/kg) via intravenosa para sedação. Foram utilizados dois hemodialisadores conectados em série, do tipo fibras ocas, baixo fluxo, membrana de polissulfona e área de 1,8m². O fluxo sangüíneo médio foi de 319,18±97,41ml/minuto, associado a um fluxo de dialisato de 500ml/min. A anticoagulação foi feita com heparina sódica em 100UI/kg para primming, repetida na dose de 53,86±18,61UI/kg/hora. Foram avaliados: tempo de coagulação, tempo de protrombina, tempo de tromboplastina parcial ativada e contagem plaquetária, e verificado trombocitopenia nos grupos dialisados. O melhor acesso vascular foi proporcionado pelo cateterismo unilateral com cateter lúmen-duplo (Grupos II e III), podendo a técnica de hemodiálise ser empregada na espécie eqüina, com dialisadores de alta eficiência, em sessão de seis horas de diálise.

Hemodialysis adequacy was studied in four groups with six horses in each: the treatments: group I animals were submitted to unilateral central venous catheter (control group); group II animals were submitted to unilateral central venous double lumen catheter and one six-hour session of hemodialysis; group III horses were submitted to unilateral central venous double lumen catheter and to two six-hour session of hemodialysis, and group IV horses were submitted to bilateral central venous mono lumen catheter and to one six-hour session of hemodialysis. Xilazine 10 percent (0.4mg/kg) and acepromazine 2 percent (0.008 mg/kg) were iv administrated for sedation. Two hollow fiber, 1.8m² low flux polyssulfone hemodialysis apparatus were used in a connected serie. The mean blood flux was 319.18±97.41ml/min with a dialisate flux of 500ml/min. Anticoagulation was performed with sodium heparin, 100UI/kg for priming at the dose of 53.86±18.61UI/kg/h. Anticoagulation monitoring was performed by clotting time, protrombin time, tromboplastin activated time, and platelet number. Decrease in platelet number was detected in groups submitted to dialysis. The best vascular access was performed with double lumen catheter and the hemodialysis may be used in equine practice, with high performance dialyze used in six- hour session.

Cyclodialysis in the treatment of nonhealing secondary glaucoma.

In cases of glaucoma with elevated intra ocular pressure, the first objective of treatment is its reduction and normalization. In cases when excessive pressure has resulted in a loss of vision, the aim of treatment is to attenuate the pain and to improve the comfort of the patient. This paper presents a case of glaucoma secondary to anterior uveitis, resistant to pharmacological treatment. A gonioscopic examination in both eyes revealed a narrowed filtration angle. An ophthalmoscopic examination of the fundus showed partial retinal detachment. The cyclodialysis procedure was performed in both eyes. In the post-surgery period, intraocular pressure (IOP) in both eyes showed a downward tendency. The treatment of glaucoma secondary to anterior uveitis, particularly in its advanced state, often requires the combination of pharmacological treatment with a surgical procedure. Positive results in maintaining the patency of the fistula were observed after the application of a steroid therapy and a regular massage of filtration bleb, which significantly contributed to maintaining IOP at a desired level.

Dialysis of boar semen prior to freezing-thawing: its effects on post-thaw sperm characteristics.

Low-molecular weight components of the seminal plasma have a detrimental effect on sperm function. The present study was undertaken to evaluate the effect of the removal of low-molecular weight components by dialysis on sperm characteristics prior to and after freezing. Semen, collected from 5 boars, was extended in Kortowo-3 extender (K-3, Poland) and cooled for 3h (control non-dialysis) or dialyzed for 5h in semi-permeable dialysis bags of 12-14kDa molecular weight cut-off prior to freezing. The semen samples were diluted in lactose-hen egg yolk-glycerol extender (lactose-HEY-G) or lactose-lyophilized lipoprotein fractions-glycerol extender (lactose-LPFo-G), packaged into aluminum tubes and frozen in a controlled programmable freezer. Pre-frozen and frozen-thawed spermatozoa were evaluated for motility, plasma membrane (SYBR-14 and propidium iodide) and acrosome integrity, mitochondrial function (Rhodamine 123) and ATP content. The results of the study showed that dialysis significantly improved the sperm characteristics prior to freezing. Dialysis enhanced (P<0.05) post-thaw sperm motility, plasma membrane integrity and mitochondrial function, but had no significant effect (P>0.05) on recovery of spermatozoa with intact acrosomes. Furthermore, dialyzed spermatozoa exhibited higher (P<0.05) ATP content compared with the control after freezing-thawing. Consistent inter-boar variability was detected mainly in dialyzed semen following freezing-thawing. These results indicated that the improvement in sperm quality characteristics prior to freezing and the post-thaw sperm recovery were due to the removal of low-molecular weight components from the seminal plasma. It can be suggested that dialysis is effective in improving the post-thaw quality of boar spermatozoa and has also great practical importance in improving the protocols for cryopreservation of semen. Dialysis may also contribute to a better understanding of different mechanisms underlying cryo-induced damage to boar spermatozoa.

Measurement of free thyroxine concentration in horses by equilibrium dialysis.

The purpose of the study reported here was to validate measurement of free thyroxine (fT4) concentration in equine serum by equilibrium dialysis (fT4D), and to compare values with fT4 concentration measured directly and with total T4 (TT4) concentration. The fT4D, fT4, and TT4 concentrations were measured over a range of values in euthyroid horses and horses made hypothyroid by administration of propylthiouracil (PTU). Concentrations of fT4D (<1.8-83 pmol/L) were consistently higher than those of fT4 (<1-40 pmol/L). There was a significant (P < .001) regression of fT4D on fT4 in 503 samples from normal horses (y = 2.086x - 0.430). In baseline samples from 71 healthy euthyroid horses, fT4 concentration ranged from 6-21 pmol/L (median, 11 pmol/L; 95% confidence interval [CI]10.5-11.8 pmol/L), and fT4D concentration ranged from 7-47 pmol/L (median, 22 pmol/L; 95% CI 20.9-25.1 pmol/L). Free T4D, fT4, and TT4 concentrations were also measured in 34 ill horses. Horses consuming PTU and ill horses had significantly (P < .05) lower serum concentration of TT4, fT4, and fT4D than did clinically normal, healthy horses. If serum samples from ill horses were further subdivided into samples from horses that lived and samples from horses that died, fT4D concentration was not significantly different in ill horses that lived, compared with that in healthy horses, whereas fT4 concentration was still significantly decreased in ill horses that died (P < 0.001). We conclude that measurement of fT4 concentration by equilibrium dialysis is a valid technique in the horse, and its use may provide improved ability to distinguish nonthyroidal illness syndrome from hypothyroidism in that species.

Measurement of serum concentrations of free thyroxine, total thyroxine, and total triiodothyronine in cats with hyperthyroidism and cats with nonthyroidal disease.

OBJECTIVE: To determine the usefulness of measuring serum free thyroxine (T4) concentration as a diagnostic test for hyperthyroidism in cats, and to determine the influence of nonthyroidal disease on free T4 concentration in cats without hyperthyroidism. DESIGN: Prospective case series. ANIMALS: 917 cats with untreated hyperthyroidism, 221 cats with nonthyroidal disease, and 172 clinically normal cats. PROCEDURE: Serum free T4, total T4, and total triiodothyronine (T3) concentrations were measured in cats with untreated hyperthyroidism and cats with nonthyroidal disease. Serum total T4 and T3 concentrations were determined by use of radioimmunoassay, and free T4 concentration was measured by use of direct equilibrium dialysis. Reference ranges for hormone concentrations were established on the basis of results from the 172 clinically normal cats. RESULTS: Sensitivity of serum free T4 concentration as a diagnostic test for hyperthyroidism was significantly higher than the test sensitivity of either total T4 or T3 concentration. Of the 221 cats with nonthyroidal disease, 14 had a high free T4 concentration (ie, false-positive result). Therefore, calculated specificity of measuring serum free T4 concentration as a diagnostic test for hyperthyroidism was significantly lower than test specificity of measuring either the total T4 or T3 concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that determination of free T4 concentration is useful in the diagnosis of hyperthyroidism, especially in cats in which hyperthyroidism is suspected but total T4 and T3 concentrations are within reference ranges. However, because some cats with nonthyroidal disease have high serum free T4 concentrations, hyperthyroidism should not be diagnosed solely on the finding of high free T4 concentration.

Inflammatory response to intramuscular implantation of polyacrylonitrile ultrafiltration probes in sheep.

Polyacrylonitrile is used in the manufacture of dialysis membranes. These membranes are fundamental to the functioning of implantable probes for microdialysis and ultrafiltration sampling of tissue fluids. Although in vivo experimentation using polyacrylonitrile has been reported to cause little inflammatory response when implanted subcutaneously, such information is not available for intramuscular implantation in sheep. The procaine and benzathine salts of penicillin are formulated for intramuscular injection. These salts of penicillin or the formulation excipients may cause inflammatory reactions. Use of polyacrylonitrile probes to draw samples from sites at which these formulations have been injected may be compromised by inflammation or direct interaction between formulation excipients and the dialysis membrane. The aim of this project was to describe tissue responses to intramuscular implantation of polyacrylonitrile in the presence and absence of either procaine or procaine plus benzathine salts of penicillin G. Each of 20 normal sheep was implanted with two ultrafiltration probes, one at the site of an injection of procaine or benzathine plus procaine penicillin G. Similar injections were also made at remote intramuscular sites. After 8, 9, and 11 days of the experiment, sheep were killed and the injection and implantation site muscle were excised and prepared for histopathological examination. The implantation of the probe alone caused greater inflammatory response than the injection of procaine or procaine plus benzathine penicillin G at remote intramuscular sites. The histopathological lesions were greatest where the implantation site was coupled with the injection of either formulation of penicillin G. Polyacrylonitrile may not be a suitable dialysis membrane material for intramuscular implantation in sheep.

Effect of dialysis before storage or cryopreservation on fertilizing ability of fowl semen.

Semen of broiler breeder males was collected and used for artificial insemination. Semen was used undiluted and diluted and both inseminated immediately after collection. Only diluted semen was used after storage. Half of the semen samples to be stored were first dialyzed. Storage of fresh, diluted semen was for 4 or 24 h at 5 C or by cryopreservation. The Beltsville Poultry Semen Extender was used for dilution and dimethylsulfoxide (DMSO) was used as cryoprotectant. Fertilizing ability was determined as the percentage fertilized eggs of eggs collected from Day 2 to 17 after a single insemination. Dialysis had a small positive effect on fertilizing ability of semen stored for 24 h. Dialysis before cryopreservation did not improve fertilizing ability. The effects of storage for 4 h were only apparent in eggs collected 9 or more d after insemination. Semen stored for 24 h gave reduced fertilizing ability in the 1st wk after insemination, especially when no dialysis was applied.

[Obtaining fecal fluid from calves: dialysis compared to filtration]. / Gewinnung von Kotwasser beim Kalb: Dialyse im Vergleich zur Filtration.

To obtain faecal fluid from the calf by dialysis the following procedure was used: small dialysis bags filled with dextran 10% were placed in the cooled (4 degrees C) faecal sample to which gentamycin was added to inhibit fermentation. Equilibrium between dialysate and surrounding fluid was reached after 24 hours. However, depending on the consistency of the faeces, after 6 hours 90 to 95% of the final osmolality at 24 hours was reached. In regard to osmolality, sodium, potassium and chloride concentrations as well as to concentrations of short chain fatty acids no significant differences could be observed between the faecal fluid obtained by dialysis or by centrifugation (3500 g, 20 min, 4 degrees C) and subsequent filtration. Distinctly higher concentrations of calcium and inorganic phosphate were measured in the filtrates; a possible explanation is that these ions in faeces are largely present as poorly diffusible compounds (in particular with faeces of a more solid consistency) or are bound to the cell membranes.

[Comparison between dialysis methods and centrifugation/filtration for collecting fecal fluid from calves]. / Vergleich zwischen Dialyseverfahren und Zentrifugation/Filtration zur Gewinnung von Kotwasser beim Kalb.

Faecal samples of different consistency from 15 calves were subjected to two methods to separate faecal fluid. Osmolality as well as the concentrations of electrolytes and short chain fatty acids were determined for each of the fluid samples obtained and the results compared. For dialysis the following procedure was used: small dialysis bags filled with dextran 10% were placed in the cooled (4 degrees C) faecal sample to which gentamycin was added to inhibit fermentation. Equilibrium between dialysate and surrounding fluid was reached after 24 hours. However, depending on the consistency of the faeces, after 6 hours 90 to 95% of the final osmolality at 24 hours was reached. In regard to osmolality, sodium, potassium and chloride concentrations as well as to concentrations of short chain fatty acids no significant differences could be observed between the faecal fluid obtained by dialysis or by centrifugation (3500 g, 20 min, 4 degrees C) and subsequent filtration. Distinctly higher concentrations of calcium and inorganic phosphate were measured in the filtrates; the probable explanation is that these ions in faeces are largely present as poorly diffusible compounds (in particular with faeces of a more solid consistency) or bound to the cell membranes.

Continuous-flow hollow-fiber dialysis system for preparation of bovine carrier erythrocytes.

Small and large molecules were encapsulated in bovine RBC by a dialysis process. Encapsulation efficiencies were approximate 30%, yielding normal carrier cells. The new process of encapsulation used hollow fibers and was a continuous-flow system. Preparation of carrier RBC by hollow-fiber dialysis was compatible with blood banking principles. The continuous-flow system was easily assembled from commercially available hollow-fiber units.

Effect of gastrointestinal parasite infection in calves on the preparation of carrier erythrocyte ghosts and on erythrocyte survival.

Calves were inoculated with helminths to induce a severe anemia. Increased erythrocyte mean cell volume caused by acute parasitism made erythrocytes more resistant to osmotic stress as reflected in the percentage of encapsulation parameters. Preparation of resealed carrier erythrocytes before and during parasitosis was studied. Low percentage of encapsulation of the marker [14C]sucrose could be corrected by extended dialysis time. Typically, 40% encapsulation was found in erythrocytes from healthy calves, whereas only 20% encapsulation was found for erythrocytes from helminth-infected animals.