Computed tomography of the coelomic cavity in healthy veiled chameleons (Chamaeleo calyptratus) and panther chameleons (Furcifer pardalis).

Background: Veiled chameleon (Chamaeleo calyptratus) and panther chameleon (Furcifer pardalis) are two of the most popular pet chameleons, and consequently, these species are frequently evaluated in veterinary practices. According to our review of the literature, normal computed tomography (CT) anatomy of these lizards has not been previously described. Aim: The purposes of this prospective study were to describe the normal CT anatomy of the coelomic organs in healthy patients and to provide normal reference values in these species. Methods: Seventeen clinically healthy veiled chameleons and 15 clinically healthy panther chameleons were included. All CT studies were performed with the chameleons under light anesthesia and positioned in sternal recumbency. Studies were performed with a 16-slice helical CT scanner with a slice thickness of 0.625 mm. The authors recorded qualitative and quantitative CT characteristics of the coelomic structures. Macroscopic cross-sectional anatomy was performed for comparison of the CT findings. Results: Heart, lungs, liver, including caudal vena cava, hepatic vessels, gallbladder, esophagus, stomach, intestines, gonads, fat bodies, kidneys, and urinary bladder could be visualized with CT. The spleen, pancreas, and adrenal glands could not be identified. Conclusion: This study provides a guide to the normal cross-sectional and computed tomographic anatomy of the coelomic cavity in veiled and panther chameleons. Our results could be used as a reference for future research studies or comparison of clinically ill patients.

Reference Range of Kaolin-Activated Thromboelastography (TEG) Values in Healthy Pet Rabbits (Oryctolagus cuniculus).

Thromboelastography (TEG) is a viscoelastic technique that allows the examination of both cellular and plasma protein clotting factors. Thromboelastography helps to investigate the underlying coagulopathy and to monitor therapeutic modalities. Although viscoelastic techniques have been used in human and veterinary medicine, reference ranges in pet rabbits are missing. The objective of this study is to establish the reference-range values of TEG parameters in healthy pet rabbits. 24 healthy pet rabbits of different breeds were included: 16 crossbreeds, four Californians, two lops, one lionhead, and one angora. Four rabbits were less than one year old and 20 were older than one year. Twelve rabbits were neutered females, 10 neutered males, and two were intact females. Health status was assessed through a physical examination, a complete blood work, and a coagulation profile. A TEG 5000 Thromboelastograph Hemostasis System was used with kaolin-activated citrated whole blood. All samples were analysed 30 min postextraction. The TEG reference ranges were reaction time (R) 1.4-6.9 min; clot formation time (K) 0.8-2.2 min; α angle 65.8-82.2 degrees; maximal amplitude (MA) 53.7-73.5 mm; measure of clot strength/firmness (G-value) 5796.6-13,885.9 dyn/cm2; and percentage of clot lysis in 30 min (LY30%) 0-41.5%. This study provides the reference ranges of TEG in pet rabbits.

Undifferentiated Wing Sarcoma in a Peach-Faced Lovebird (Agapornis roseicollis).

A 10-year-old peach-faced lovebird (Agapornis roseicollis) was evaluated for an ulcerated and painful mass at the location of a fracture 2 years previously. Whole body radiographs showed a humeral fracture with a presumptive neoplastic proliferation in the distal diaphysis. Right wing amputation was elected but the animal died during recovery from surgery. Histopathological examination of the amputated wing revealed an infiltrative sarcomatous neoplastic proliferation. Immunohistochemistry (IHC) was carried out to characterize the tumour using antibodies against vimentin, desmin, smooth muscle actin (SMA), S-100, ionized calcium-binding adapter molecule-1 (IBA-1), CD18, cytokeratin and epithelial membrane antigen (EMA). The mesenchymal component of the mass was immunolabelled for vimentin and SMA and sparse epithelial cells were immunopositive for cytokeratin. Very few scattered cells were immunopositive for CD18 and IBA-1. The final diagnosis was consistent with an undifferentiated sarcoma with intralesional hyperplastic epithelium. According to the location, the history of a previous fracture and the histological pattern and IHC profile, the tumour was classified as an undifferentiated sarcoma with entrapped air sac epithelium.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and humoral responses against different variants of concern in domestic pet animals and stray cats from North-Eastern Spain.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of the coronavirus disease 2019 (COVID-19) pandemic in humans, is able to infect several domestic, captive and wildlife animal species. Since reverse zoonotic transmission to pets has been demonstrated, it is crucial to determine their role in the epidemiology of the disease to prevent further spillover events and major spread of SARS-CoV-2. In the present study, we determined the presence of virus and the seroprevalence to SARS-CoV-2, as well as the levels of neutralizing antibodies (nAbs) against several variants of concern (VOCs) in pets (cats, dogs and ferrets) and stray cats from North-Eastern of Spain. We confirmed that cats and dogs can be infected by different VOCs of SARS-CoV-2 and, together with ferrets, are able to develop nAbs against the ancestral (B.1), Alpha (B.1.1.7), Beta (B.1.315), Delta (B.1.617.2) and Omicron (BA.1) variants, with lower titres against the latest in dogs and cats, but not in ferrets. Although the prevalence of active SARS-CoV-2 infection measured as direct viral RNA detection was low (0.3%), presence of nAbs in pets living in COVID-19-positive households was relatively high (close to 25% in cats, 10% in dogs and 40% in ferrets). It is essential to continue monitoring SARS-CoV-2 infections in these animals due to their frequent contact with human populations, and we cannot discard the probability of a higher animal susceptibility to new potential SARS-CoV-2 VOCs.

Clinical, imaging, and pathologic features in cases of neurologic disease in 3 psittacine birds.

We used magnetic resonance imaging (MRI) to evaluate the CNS, and confirmed CNS lesions histologically, in 3 psittacine birds with neurologic signs. One bird was recumbent as a result of non-ambulatory paraparesis, and 2 birds were ataxic with impaired proprioception. In all 3 cases, imaging was performed, and infectious diseases were excluded in cases 1 and 2. In case 1, a large mass arose from the left lung; in case 2, a multinodular coelomic mass encompassed the left caudal pulmonary area to the left cranial renal pole; and in case 3, a diffuse hyperintensity affected the lumbar spinal cord. In the first 2 cases, masses invaded the vertebral canal, causing spinal cord compression. All 3 birds were euthanized given the poor prognosis, and postmortem examinations were performed. The final diagnoses were pulmonary adenocarcinoma in cases 1 and 2, and granulomatous and lymphocytic leptomeningitis caused by Mycobacterium genavense in case 3. MRI enabled visualization of the lesions in the affected area of the CNS, and MRI findings were confirmed by histopathology.

Health status of free-ranging pure and cross-mixed miniature swine population from Northeast Spain.

BACKGROUND: Miniature pigs have gained popularity as companion animals in the recent years in Spain. Due to the fact that their abandonment and crossing breeds with wild boars can cause severe problems, investigation about the health status is needed. OBJECTIVES: The aim of this study was to determine their health status according to the clinical findings during physical examination and the results of antibody serology tests against selected infectious diseases. METHODS: Two-hundred and eleven miniature pigs (Sus scrofa) were included in the study. Their origin, age, sex, housing conditions and diet were recorded. RESULTS: The housing of the animals ranged from wild animals to ones living in animal sanctuaries. The diet varied from a natural one in the wild to commercial and homemade food. Thirty animals out of two-hundred and eleven were hybrids between miniature pigs and wild boars according to morphological characteristics. Antibody serology techniques of Influenza A virus, Hepatitis E virus, brucellosis, tuberculosis, African swine fever, Classical swine fever and Aujeszky's disease was performed. The prevalence for Influenza A was 5.30%, for Hepatitis E was 5.35% and the rest tested negative. It is important to control and monitor these zoonotic infections to prevent Public Health problems. CONCLUSIONS: The results obtained from this investigation demonstrated that the animals' health status in this study is optimal and the diseases prevalence is similar or minor when compared to previous studies. This study confirms the hybridization of miniature pig and wild boar in Catalonia.

Vertebral heart score to evaluate cardiac size in thoracic radiographs of 124 healthy rats (Rattus norvegicus).

Dilated cardiomyopathy is a relatively common disease in pet rats (Rattus norvegicus); however, there is a lack of radiographic references for the normal cardiac size in this species. The aim of this prospective, anatomical and reference interval study was to establish quantitative radiographic reference range measurements for the vertebral heart score (VHS) in rats. Right lateral (RL), ventrodorsal (VD), and dorsoventral (DV) radiographs of clinically healthy rats (n = 124) were evaluated. Measurements were performed by 2 expert readers who were unaware of signalment data. The mean values and references intervals of VHS were 7.7 and 7.0-8.5 for the RL, 7.5 and 6.6-8.6 for the VD, and 7.9 and 6.9-9.0 for the DV, with VHS values greater in males than in females. The measurements reported in this study can be used by the clinician as an objective tool to evaluate cardiac size in rats, in order to improve the diagnosis and treatment of cardiac diseases.

Paradoxical Vestibular Syndrome Caused by a Presumptive Cerebellar Infarction in a Rabbit.

A 6-year-old, female spayed rabbit (Oryctolagus cuniculus) presented with right paradoxical vestibular signs. Magnetic resonance imaging was performed and findings were consistent with an ischemic infarct of the cerebellum. The patient improved gradually and was free of clinical signs at the time this article was written. To the authors' knowledge this is the first case report of a paradoxical vestibular syndrome in a rabbit secondary to a presumptive ischemic infarct. Strokes should be included in the differential diagnosis of central vestibular syndrome in rabbits.

B Cell-Derived Extracellular Vesicles Reveal Residual B Cell Activity in Kidney Graft Recipients Undergoing Pre-Transplant Desensitization.

Background: Living-donor kidney transplant (LDKT) recipients undergoing desensitization for Human Leukocyte Antigen (HLA)-incompatibility have a high risk of developing antibody-mediated rejection (ABMR). The purpose of the study is to evaluate if residual B cell activity after desensitization could be estimated by the presence of circulating B cell-derived extracellular vesicles (BEVs). Methods: BEVs were isolated by Sepharose-based size exclusion chromatography and defined as CD19+ and HLA-II+ extracellular vesicles. We analyzed stored serum samples from positive crossmatch LDKT recipients before and after desensitization at first post-transplant biopsy and at 12-month protocol biopsy (n = 11). Control groups were formed by hypersensitized patients who were not submitted to desensitization (n = 10) and by low-risk recipients (n = 9). A prospective validation cohort of 11 patients also included the analysis of B cells subpopulations in recipients' blood and lymph nodes recovered upon graft implantation, along with BEVs analysis before and after desensitization. Results: We found out that CD19+ and HLA-II+BEVs dropped significantly after desensitization and relapse in patients who later developed ABMR was evident. We validated these findings in a proof-of-concept prospective cohort of 6 patients who received the same desensitization protocol and also in a control group of 5 LDKT recipients. In these patients, B cell subpopulations were also studied in recipients' blood and lymph nodes that were recovered before the graft implantation. We confirmed the significant drop in BEVs after desensitization and that this paralleled the reduction in CD19+cells in lymph nodes, while in peripheral blood B cells, this change was almost undetectable. Conclusions: BEVs reflected B cell residual activity after desensitization and this could be a valid surrogate of humoral alloreactivity in this setting.

External ear canal mycobiome of some rabbit breeds.

The genus Malassezia is part of the normal skin mycobiota of a wide range of warm-blooded animals. In this genus, M. cuniculi is the only species described from rabbits. However, Malassezia species are rarely studied in lagomorphs. In the present study, the presence of Malassezia was assessed in samples from the external ear canal of healthy rabbits of different breeds. Cytological and culture techniques, Sanger sequencing, and Next-generation sequencing (NGS) were used to describe the ear mycobiota in the samples. Although no growth was observed in the cultured plates, cytological examination revealed the presence of round cells similar to those of Malassezia yeasts. For metagenomics analysis, the D1/D2 domain of the large subunit of the ribosomal DNA (LSU rDNA) was PCR amplified and the resulting reads were mapped against a custom-made cured database of 26S fungal sequences. NGS analysis revealed that Basidiomycota was the most abundant phylum in all the samples followed by Ascomycota. Malassezia was the most common genus presenting the highest abundance in the external ear canal. Malassezia phylotype 131 and M. cuniculi were the main sequences detected in the external auditory canal of rabbits. The study included both lop-eared and erect-eared rabbits and no differences were observed in the results when comparing both groups. This is the first attempt to study the external ear canal mycobiome of rabbits of different breeds using NGS. LAY SUMMARY: In the present study, the presence of Malassezia was assessed in samples from the external ear canal of healthy rabbits of different breeds. Cytological and culture techniques, Sanger sequencing, and Next-generation sequencing (NGS) were used to describe the ear mycobiota in the samples.

A Europe wide acceptable mismatch program will enable transplantation of long waiting highly sensitised patients with a compatible donor.

Immunisation against Human Leucocyte Antigens (HLA) can be caused by pregnancy, blood transfusion, or organ transplants. The HLA antibody status of a given patient significantly influences their access and waiting time to transplant. For some highly sensitised patients (HSP) there is hardly any suitable donor available in the deceased donor pool of their allocation organisation and therefore they wait a very long time before being offered a kidney for transplant. Especially patients with rare HLA phenotypes in relation to the actual donor pool are waiting extremely long. As HLA phenotypes are different in the various European populations, we hypothesized that extension of the donor pool outside the respective allocation system will increase the chance of receiving a compatible transplant for this subgroup of highly sensitised patients. One of the objectives of the EUROSTAM project, (a Europe-wide Strategy to enhance Transplantation of highly sensitised patients on the basis of Acceptable HLA Mismatches) was to develop a tool to compare the chance of transplanting HSP in different European populations with donor organs from within and outside their own donor pool. Information on the HLA type and ABO blood group of the actual donor population, as well as the acceptable mismatches of long waiting HSP were obtained from the EUROSTAM partner organizations i.e. Eurotransplant (ET), UK National Health Service Blood and Transplant (NHSBT), Barcelona, Prague and Athens. Results from simulations using the newly developed tool shows that 195 (27%) of the 724 long waiting highly sensitised patients registered at each partner organisation have increased chances of transplant in a different European donor pool. This makes a strong case for sharing kidneys between European countries for selected difficult to transplant patients.

Detection of antibodies to denatured human leucocyte antigen molecules by single antigen Luminex.

The anti-HLA antibody detection has been improved in sensitivity and specificity with solid-phase antigen bead (SAB) assays based on Luminex. However, false positive results due to denatured HLA (dHLA) may arise after single antigen test. The aim of this study was to compare the performance of the two Luminex technology-based anti-HLA detection kits available in the market in showing undesired anti-HLA antibody results. A prospective cohort was assessed for anti-HLA antibodies with single antigen A manufacturer (AM) kit and a comparison cohort with single antigen B manufacturer (BM) kit. A total of 11 out of 90 patients in a prospective cohort presented monospecific HLA-I antibodies with AM, and 5 out of 11 confirmed monospecific reaction with BM. Despite the confirmation of monospecific reaction with both manufacturers, 80% were assigned as dHLA reaction by specific crossmatch. Further comparative cohorts detected four out of six monospecific reactions with BM that were confirmed as possible dHLA reactions. A positive SAB test should rule out a reaction against a dHLA molecule, thus avoidance of prolonged waitlist periods or misattribution of anti-HLA reactions after transplantation.

Use of De Novo mTOR Inhibitors in Hypersensitized Kidney Transplant Recipients: Experience From Clinical Practice.

BACKGROUND: It is commonly believed that mTOR inhibitors (mTORi) should not be used in high-immunological risk kidney transplant recipients due to a perceived increased risk of rejection. However, almost all trials that examined the association of optimal-dose mTORi with calcineurin inhibitor (CNI) have excluded hypersensitized recipients from enrollment. METHODS: To shed light on this issue, we examined 71 consecutive patients with a baseline calculated panel reactive antibody (cPRA) ≥50% that underwent kidney transplantation from June 2013 to December 2016 in our unit. Immunosuppression was based on CNI (tacrolimus), steroids and alternatively mycophenolic acid (MPA; n = 38), or mTORi (either everolimus or sirolimus, n = 33, target trough levels 3-8 ng/mL). RESULTS: Demographic and immunological risk profiles were similar, and almost 90% of patients in both groups received induction with lymphocyte-depleting agents. Cox-regression analysis of rejection-free survival revealed better results for mTORi versus MPA in terms of biopsy-proven acute rejection (hazard ratio [confidence interval], 0.32 [0.11-0.90], P = 0.031 at univariable analysis and 0.34 [0.11-0.95], P = 0.040 at multivariable analysis). There were no differences in 1-year renal function, Banff chronicity score at 3- and 12-month protocol biopsy and development of de novo donor-specific antibodies. Tacrolimus trough levels along the first year were not different between groups (12-mo levels were 8.72 ± 2.93 and 7.85 ± 3.07 ng/mL for MPA and mTORi group respectively, P = 0.277). CONCLUSIONS: This single-center retrospective cohort analysis suggests that in hypersensitized kidney transplant recipients receiving tacrolimus-based immunosuppressive therapy similar clinical outcomes may be obtained using mTOR inhibitors compared to mycophenolate.

Successful use of nonantigen-specific immunoadsorption with antihuman Ig-columns in kidney graft antibody-mediated rejection.

INTRODUCTION: Nonantigen-specific immunoadsorption (IA) has proven to be effective in acute antibody-mediated rejection (aAMR). However, there is a lack of solid studies evaluating the safety and efficacy of IA with antihuman Ig-columns in aAMR. For chronic-active AMR (cAMR), no studies have evaluated the efficacy of nonantingen-specific IA with antihuman Ig-columns. The purpose of this study was to evaluate the role of nonantigen-specific IA with antihuman Ig-columns in the treatment of both aAMR and cAMR in kidney transplantation. MATERIAL AND METHODS: In retrospective and observational study, kidney graft and recipient survival rates were assessed after treatment of aAMR and cAMR with nonantigen-specific IA with Ig-Flex columns (Therasorb) between January 2012 and May 2018. Protocols included nonantigen-specific IA, rituximab, intravenous immunoglobulin, and rescue plasma exchange, if necessary. RESULTS: The study included 14 patients with AMR (acute in 9, chronic active in 5). For aAMR, mean follow-up was 13 ± 6 months, and patient and graft survival were, respectively, of 100% and 83%, with a mean increase in estimated glomerular filtration rate (eGFR) of 7.98 ± 12.96, 10.18 ± 16.71, and 11.43 ± 13.85 mL/min/1.72 m2 (P > .05) at 3, 12 months after treatment, and at the end of follow-up, respectively. For cAMR, mean follow-up was 14 ± 8 months, and patient and graft survival were, respectively, of 100% and 60%, with an average increase in eGFR of 4.30 ± 7.86, 5.64 ± 10.47, and 14.5 ± 7.86 mL/min/m2 (P > .05) at 3, 12 months after IA treatment, and at the end of the follow-up, respectively, although 40% did not respond and required chronic hemodialysis. CONCLUSION: Nonantigen-specific IA with Ig-Flex columns was safe and effective for aAMR treatment in kidney transplantation. In cAMR, IA with Ig-Flex columns was associated with a satisfactory kidney graft survival, suggesting that IA could potentially offer some benefits supporting its indication in cAMR.

Donor/Recipient HLA Molecular Mismatch Scores Predict Primary Humoral and Cellular Alloimmunity in Kidney Transplantation.

Donor/recipient molecular human leukocyte antigen (HLA) mismatch predicts primary B-cell alloimmune activation, yet the impact on de novo donor-specific T-cell alloimmunity (dnDST) remains undetermined. The hypothesis of our study is that donor/recipient HLA mismatches assessed at the molecular level may also influence a higher susceptibility to the development of posttransplant primary T-cell alloimmunity. In this prospective observational study, 169 consecutive kidney transplant recipients without preformed donor-specific antibodies (DSA) and with high resolution donor/recipient HLA typing were evaluated for HLA molecular mismatch scores using different informatic algorithms [amino acid mismatch, eplet MM, and Predicted Indirectly Recognizable HLA Epitopes (PIRCHE-II)]. Primary donor-specific alloimmune activation over the first 2 years posttransplantation was assessed by means of both dnDSA and dnDST using single antigen bead (SAB) and IFN-γ ELISPOT assays, respectively. Also, the predominant alloantigen presenting pathway priming DST alloimmunity and the contribution of main alloreactive T-cell subsets were further characterized in vitro. Pretransplantation, 78/169 (46%) were DST+ whereas 91/169 (54%) DST-. At 2 years, 54/169 (32%) patients showed detectable DST responses: 23/54 (42%) dnDST and 31/54 (57%) persistently positive (persistDST+). 24/169 (14%) patients developed dnDSA. A strong correlation was observed between the three distinct molecular mismatch scores and they all accurately predicted dnDSA formation, in particular at the DQ locus. Likewise, HLA molecular incompatibility predicted the advent of dnDST, especially when assessed by PIRCHE-II score (OR 1.014 95% CI 1.001-1.03, p=0.04). While pretransplant DST predicted the development of posttransplant BPAR (OR 5.18, 95% CI=1.64-16.34, p=0.005) and particularly T cell mediated rejection (OR 5.33, 95% CI=1.45-19.66, p=0.012), patients developing dnDST were at significantly higher risk of subsequent dnDSA formation (HR 2.64, 95% CI=1.08-6.45, p=0.03). In vitro experiments showed that unlike preformed DST that is predominantly primed by CD8+ direct pathway T cells, posttransplant DST may also be activated by the indirect pathway of alloantigen presentation, and predominantly driven by CD4+ alloreactive T cells in an important proportion of patients. De novo donor-specific cellular alloreactivity seems to precede subsequent humoral alloimmune activation and is influenced by a poor donor/recipient HLA molecular matching.

Ultrasonographic appearance of the coelomic cavity organs in healthy veiled chameleons (Chamaeleo calyptratus) and panther chameleons (Furcifer pardalis).

Veiled chameleons (Chamaeleo calyptratus) and panther chameleons (Furcifer pardalis) are the most popular chameleons over the world, and consequently, two of the most frequent species presenting to veterinary practices. However, published studies on normal ultrasonographic anatomy for these lizards are currently lacking. The objectives of this prospective anatomic study were to develop an ultrasound protocol for evaluation of the coelomic cavity in these species and describe the normal ultrasonographic anatomy of the coelomic organs. Seventeen healthy veiled chameleons and 15 healthy panther chameleons were included. A linear 18 MHz transducer was used. Chameleons were sedated and restrained in right lateral recumbency by an assistant. Longitudinal and transverse images were acquired, and authors recorded qualitative and quantitative ultrasonographic characteristics of the coelomic structures. The kidneys, liver, caudal vena cava, hepatic veins, portal vein, gallbladder, wall of the stomach and intestine, gonads and, when distended, urinary bladder could be visualized during ultrasonography of the coelomic cavity in both species. The spleen, pancreas, and adrenal glands could not be identified. Findings from the current study supported the use of ultrasonography for veiled chameleons (Chamaeleo calyptratus) and panther chameleons (Furcifer pardalis) with suspected intracoelomic diseases and provided normal reference information for future studies of these chameleon species.

Impact of Discards for Living Donor Kidney Transplantation in a Transplant Program.

OBJECTIVE: Living donor kidney transplantation (LDKT) is the best treatment for end-stage renal disease. In this setting, a significant percentage of transplants are not undertaken because of medical and nonmedical reasons of both donors and recipients. However, the impact of these discards in a transplant program has not been identified thoroughly so far. Our objective was to clarify key reasons for exclusion of LDKTs and the consequences for the discarded transplant candidates in the following 5 years. METHODS: Analysis of donors' and recipients' characteristics of 781 couples evaluated in our hospital from January 2005 to December 2013. The consequences of discards in transplant candidates were analyzed in the cohort 2012 to 2013 (n = 106) and followed up until October 2018. RESULTS: In our study group, 402 (51.5%) LDKT couples were successfully donated, and 379 (48.5%) were excluded. Donor and transplant recipient candidates discarded were older at the evaluation (55.07 ± 12.14 years vs 51.73 ± 10.93 years, P < .001; 48.81 ± 14.05 years vs 44.62 ± 13.91 years, P < .001, respectively). The most frequent reason for kidney discard was medical contraindication found in the potential donor (47.5%; low eGFR, diabetes mellitus, impaired glucose tolerance, high blood pressure, cardiovascular pathology casually found during evaluation, and proteinuria). Of the discarded candidates from 2012 to 2013, 36.8% received a deceased donor kidney transplant, 17% a LDKT with another donor, 7.5% stayed on the waiting list, 18.9% died, 3.8% were excluded from the waiting list, and 14.2% were lost to follow-up. CONCLUSIONS: In most cases, transplantation was not undertaken because of donor pathology. Fifty-three percent of the discarded patients were eventually transplanted, with a 31.4% probability to receive an organ from another living donor.

Towards uniformity in the definition of acceptable mismatches for highly sensitized patients.

The Eurotransplant (The Eurotransplant International Foundation) acceptable mismatch programme has been shown to be a successful tool to enhance transplantation of highly sensitized patients(HSPs). However, patients with rare HLA phenotypes in relation to the Eurotransplant donor population remain on the waiting list. EUROSTAM is an European Union funded project to explore the feasibility of a Europe-wide acceptable mismatch programme enabling transplantation of HSPs with rare HLA phenotypes within their own organ exchange organization. The present study, which forms part of the EUROSTAM project, assesses the differences in the practices of the laboratories in different countries with respect to their HLA antibody profiling and risk adverseness. In the serum exchange exercises of 18 samples, a high level of variability has been shown in both assays and interpretation of results. In the data exchange exercise when all participants were given the same Luminex raw data for analysis, a high degree of consensus was reached where the median fluorescent intensity values of beads were <500 and >2000 for standard single antigen bead assays, or <500 and >5000 for assignment of acceptable mismatches. The risk adverseness analysis has showed distinct patterns of attitudes towards the perceived risks based on HLA antibody assay results, most probably influenced by the local protocols of the clinical transplant programme of each laboratory. In order to ensure fairness and maintain consistencies of organ exchange among partner transplant centres, a centralized facility will be instrumental for a uniform definition of acceptable mismatches.

Description of digital fluoroscopic excretory urography in healthy New Zealand rabbits (Oryctolagus cuniculus).

Disorders of the kidneys and urinary tract are relatively common in rabbits. Survey radiographs and abdominal ultrasound have inherent limitations in the evaluation of the urinary tract in this species; because of that, contrast study modalities are a valuable diagnostic tool in rabbits with urinary disorders. The aims of this study were to describe digital fluoroscopic excretory urography (DFEU) and to determine the urogram phases in healthy New Zealand rabbits. The median time of beginning of the nephrographic and pyelographic phases was 10 seconds (IQR: 10-12 seconds) and 1.39 minutes (IQR: 1.27-1.60 minutes). The median time in which the contrast medium was visible in the urinary bladder was 1.58 minutes (IQR: 1.44-1.72 minutes). Results of this study indicate that DFEU provides a good degree of opacification of the kidneys as well as the ureters, and allows the evaluation of the urogram phases. Text not justifed in the proof version.

Combining Sensitive Crossmatch Assays With Donor/Recipient Human Leukocyte Antigen Eplet Matching Predicts Living-Donor Kidney Transplant Outcome.

Introduction: Despite the different assays available for immune-risk stratification before living-donor kidney transplantation (LDKT), the precise type and number of tests to perform remain uncertain. Methods: In a cohort of 330 consecutive LDKT patients, all of which were complement-dependent cytotoxicity (CDC)-crossmatch negative, we retrospectively analyzed the impact on main clinical outcomes of most sensitive immunoassays (complement-dependent cytotoxicity-panel-reactive antibody [CDC-PRA], flow cytometry crossmatch [FC-XM], donor-specific antibodies [DSAs], and their complement-binding capacity DSA-C3d]), together with donor/recipient HLA eplet matching. Mean follow-up was 67 months (range 24-190 months). Results: Of 330 patients, 35 (11%) showed a CDC-PRA >20%; 17 (5%) FC-XM+; 30 (9%) DSA+, 18(5%) DSA-C3d+, with low overlapping results (10 patients positive in all donor-specific tests). Unlike HLA allele compatibility, the mean number of HLA class II eplet mismatches was higher in LDKT patients with positive baseline test results. DSA-C3d+ showed higher mean fluorescence intensity (MFI) DSA, with a cut-off MFI of 6192 accurately predicting complement fixation (area under the curve = 0.85, P = 0.008). Although all assays were associated with acute rejection (AR), only DSA-C3d+ (odds ratio [OR] = 6.64, P = 0.038) or high MFI-DSA (OR = 7.54, P = 0.038) independently predicted AR. Likewise, poorly HLA class II eplet-matched patients were at higher risk for AR, particularly patients with negative baseline test results (OR = 1.14, P = 0.019). Finally, previous AR and FC-XM+/DSA+, regardless of C3d positivity, independently predicted graft loss. Conclusion: Combining FC-XM and solid-phase assays with the evaluation of donor/recipient HLA eplet mismatches, are most accurate tools for immune-risk stratification prior LDKT.