Drug product attributes predict clinical efficacy in betibeglogene autotemcel gene therapy for ß-thalassemia.

Ex vivo autologous hematopoietic stem cell lentiviral-based gene therapy with betibeglogene autotemcel has been studied in patients with transfusion-dependent ß-thalassemia in Phase III clinical trials (HGB-207 and HGB-212), with 90% of patients reaching transfusion independence (TI). Here, we explore manufacturing parameters, drug product quality attributes, and limited patient characteristics that had an impact on clinical efficacy in HGB-207 and HGB-212. Retrospective analysis revealed that the peripheral blood vector copy number (VCN) was related to TI, with a strong correlation between peripheral blood VCN at 6 months and gene therapy-derived therapeutic protein (HbAT87Q) expression at 6 months (correlation coefficient, 0.8681; p < 0.0001; R2 = 0.7536). A peripheral blood VCN threshold of ≥0.75 copies per diploid genome at 6 months post betibeglogene autotemcel infusion provided a stringent surrogate biomarker for TI and was used as the outcome variable for multivariate analysis using a random forest classifier. The top predictive feature of clinical efficacy was found to be the percentage of lentiviral vector-positive cells in the drug product. This retrospective analysis is critical to understanding the key product quality attributes that can predict clinical efficacy in lentiviral vector gene therapy within this clinical trial population.

ACVR and ECVDI consensus statement for the standardization of the abdominal ultrasound examination.

This consensus statement is designed to provide a standard of care document and describes the ACVR and ECVDI definition for performing a standard abdominal ultrasound examination in dogs and cats. The ACVR and ECVDI define a standard abdominal ultrasonographic examination as a complete exam of the abdominal organs which is appropriately documented. The consensus statement intends to provide guidance to veterinary sonographers and veterinarians for the performance and documentation of high-quality diagnostic ultrasound examination. The document may also serve as a teaching aid for veterinary students, veterinarians, and residents in diagnostic imaging who seek proficiency in diagnostic ultrasound. Finally, it may serve an additional role in educating the public as to what a high-quality abdominal ultrasound examination should entail.

Diagnostic imaging for the assessment of acquired abdominal vascular diseases in small animals: A pictorial review.

Advances in interventional radiology and surgical techniques now allow complex abdominal diseases to be more successfully treated in small animals. Abdominal vascular alterations, acquired as individual process or as complication of other lesions such as neoplasia, can be life-threatening or at least greatly limit curative interventions of underlying diseases. Computed tomography (CT) and high-definition ultrasonography are now readily available in veterinary referral centers. Yet, there is little information currently available on the use of these modalities for the diagnosis and characterization of these vascular alterations. The purpose of this article is to review the CT and ultrasonographic findings of acquired vascular diseases in the abdomen of dogs and cats, using both the veterinary and human medicine literature as references, and highlighting essential concepts through figures.

Ultrasonographic and clinicopathological features of pyloroduodenal adenomatous polyps in cats.

Objectives The aim of the study was to report the ultrasonographic and clinicopathological findings in cats with confirmed pyloroduodenal adenomatous polyps. Methods Clinicopathological data, ultrasonographic and histopathological findings were collected retrospectively from medical records. Results Pyloroduodenal polyps appeared as small moderately echogenic and homogeneous nodules filling most of the proximal duodenal or pyloroduodenal lumen. The most common presenting signs in this study included acute vomiting and anorexia in 4/6 cats and lethargy in 3/6 cats. Two cats presented with severe anemia, suggestive of active bleeding. One cat presented with increased bilirubin without anemia, suggestive of impaired bile flow. Five cats survived surgical removal of the polyp and were free of clinical signs 817 days after the procedure. Conclusions and relevance Pyloroduodenal polyps have subtle ultrasonographic changes that can easily be mistaken for ingesta. They are characterized by a discrete small (up to 1.5 cm in diameter) homogeneous echogenic nodule filling the pyloroduodenal lumen. Pyloroduodenal polyps are benign lesions but can cause severe clinical signs including gastrointestinal bleeding or biliary obstruction. The prognosis is excellent with surgical removal.

Ileocecocolic volvulus in a German shepherd dog.

This report describes an ileocecocolic volvulus in a German shepherd dog with risk factors of previous abdominal surgeries and concurrent chronic enteropathy. Contrast-enhanced computed tomography (CT) with multiplanar reformatting was more sensitive than abdominal radiographs or ultrasound to obtain a diagnosis, because of the presence of a "whirl-sign" on CT. A combination of colopexy and cecopexy was succesfully used to treat the patient's condition.

Volvulus iléo-caeco-colique chez un Berger Allemand. Ce rapport de cas décrit un volvulus iléo-caeco-colique chez un Berger allemand ayant différents facteurs de risque incluant des chirurgies abdominales préalables à sa présentation et une entéropathie chronique. L'observation d'un 'whirl-sign' lors des reconstructions de l'examen tomodensitométrique a permis d'obtenir le diagnostic de volvulus et s'est révélé dans ce cas être plus sensible que les examens d'imagerie usuels, soit la radiographie ou l'échographie abdominale. Le traitement chirurgical a été effectué avec succès grâce à la combinaison d'une cecopexie et d'une colopexie.(Traduit par les auteurs).

Maximizing recombinant human serum albumin production in a Mut(s) Pichia pastoris strain.

Human serum albumin (HSA) is a cysteine rich molecule that is most abundant in human blood plasma. To remain viable in the market due to lower marketing costs for HSA, it is important to produce a large quantity in an economical manner by recombinant technology. The objective of this study was to maximize recombinant HSA (rHSA) production using a Mut(s) Pichia pastoris strain by fermentation process optimization. We evaluated the impact of process parameters on the production of rHSA, including induction cell density (wet cell weight, g/L) and the control of specific growth rate at induction. In this study, we demonstrated that induction cell density is a critical factor for high level production of rHSA under controlled specific growth rate. We observed higher specific productivities at higher induction cell densities (285 g/L) and at lower specific growth rates (0.0022-0.0024/h) during methanol induction phase, and achieved the broth titer of rHSA up to 10 g/L. The temperature shift from 24 to 28(o) C was effective to control the specific growth rate at low level (≤0.0024/h) during methanol induction phase while maintaining high specific productivity [0.0908 mgrHSA /(gwcw h)].

Radiographic diagnosis of mechanical obstruction in dogs based on relative small intestinal external diameters.

Mechanical obstruction is a frequent cause of acute vomiting in dogs requiring prompt diagnosis to improve patient management and prognosis. The purpose of this retrospective study was to compare small intestinal radiographic characteristics in dogs with versus without mechanical intestinal obstruction. Fifty dogs with gastrointestinal clinical signs and abdominal radiographs were recruited from hospital record archives and assigned to groups (group 1, obstructive, n = 25; group 2, nonobstructive n = 25). Abdominal radiographs were randomized and independently interpreted by three examiners who were unaware of group status. Intestinal dilation was subjectively scored based on distribution (segmental, regional or diffuse), and severity (absent, mild, moderate or severe). Small intestinal maximal diameter (SImax), L5 vertebral body height, small intestinal minimal diameter (SImin), and an estimated average of small intestinal diameters (SIave) were measured and three ratios were calculated: SImax/L5, SImax/SImin, and SImax/SIave. Segmental dilation was more prevalent in obstructed dogs for all examiners (P ≤ 0.03) and most nonobstructed dogs had no dilation (P ≤ 0.05). All ratios were higher in obstructed dogs (P < 0.002). Subjective dilation scores and ratio measurements had low interobserver agreement (absent to fair, with kappa values between -0.06 and 0.57) and reproducibility (coefficients of 0.35-0.61). Findings indicated that dogs with SImax/L5 ≤ 1.4, SImax/SImin ≤ 2, and SImax/SIave ≤ 1.3 values are very unlikely to be mechanically obstructed; dogs with SImax/L5 ≥ 2.4, SImax/SImin ≥ 3.4 and SImax/SIave ≥ 1.9 are very likely obstructed, particularly if segmental dilation (less than 25% of the small intestine) is present. Dogs with ratios falling between these thresholds may need further testing unless other signs justify surgical exploration or endoscopy.

Computed tomographic characteristics of collateral venous pathways in dogs with caudal vena cava obstruction.

Collateral venous pathways develop in dogs with obstruction or increased blood flow resistance at any level of the caudal vena cava in order to maintain venous drainage to the right atrium. The purpose of this retrospective study was to describe the sites, causes of obstruction, and configurations of venous collateral pathways for a group of dogs with caudal vena cava obstruction. Computed tomography databases from two veterinary hospitals were searched for dogs with a diagnosis of caudal vena cava obstruction and multidetector row computed tomographic angiographic (CTA) scans that included the entire caudal vena cava. Images for each included dog were retrieved and collateral venous pathways were characterized using image postprocessing and a classification system previously reported for humans. A total of nine dogs met inclusion criteria and four major collateral venous pathways were identified: deep (n = 2), portal (n = 2), intermediate (n = 7), and superficial (n = 5). More than one collateral venous pathway was present in 5 dogs. An alternative pathway consisting of renal subcapsular collateral veins, arising mainly from the caudal pole of both kidneys, was found in three dogs. In conclusion, findings indicated that collateral venous pathway patterns similar to those described in humans are also present in dogs with caudal vena cava obstruction. These collateral pathways need to be distinguished from other vascular anomalies in dogs. Postprocessing of multidetector-row CTA images allowed delineation of the course of these complicated venous pathways and may be a helpful adjunct for treatment planning in future cases.

Validation of a novel technique for creating simulated radiographs using computed tomography datasets.

Understanding radiographic anatomy and the effects of varying patient and radiographic tube positioning on image quality can be a challenge for students. The purposes of this study were to develop and validate a novel technique for creating simulated radiographs using computed tomography (CT) datasets. A DICOM viewer (ORS Visual) plug-in was developed with the ability to move and deform cuboidal volumetric CT datasets, and to produce images simulating the effects of tube-patient-detector distance and angulation. Computed tomographic datasets were acquired from two dogs, one cat, and one horse. Simulated radiographs of different body parts (n = 9) were produced using different angles to mimic conventional projections, before actual digital radiographs were obtained using the same projections. These studies (n = 18) were then submitted to 10 board-certified radiologists who were asked to score visualization of anatomical landmarks, depiction of patient positioning, realism of distortion/magnification, and image quality. No significant differences between simulated and actual radiographs were found for anatomic structure visualization and patient positioning in the majority of body parts. For the assessment of radiographic realism, no significant differences were found between simulated and digital radiographs for canine pelvis, equine tarsus, and feline abdomen body parts. Overall, image quality and contrast resolution of simulated radiographs were considered satisfactory. Findings from the current study indicated that radiographs simulated using this new technique are comparable to actual digital radiographs. Further studies are needed to apply this technique in developing interactive tools for teaching radiographic anatomy and the effects of varying patient and tube positioning.

A posteriori comparison of natural and surgical destabilization models of canine osteoarthritis.

For many years Canis familiaris, the domestic dog, has drawn particular interest as a model of osteoarthritis (OA). Here, we optimized the dog model of experimental OA induced by cranial cruciate ligament sectioning. The usefulness of noninvasive complementary outcome measures, such as gait analysis for the limb function and magnetic resonance imaging for structural changes, was demonstrated in this model. Relationships were established between the functional impairment and the severity of structural changes including the measurement of cartilage thinning. In the dog model of naturally occurring OA, excellent test-retest reliability was denoted for the measurement of the limb function. A criterion to identify clinically meaningful responders to therapy was determined for privately owned dogs undergoing clinical trials. In addition, the recording of accelerometer-based duration of locomotor activity showed strong and complementary agreement with the biomechanical limb function. The translation potential of these models to the human OA condition is underlined. A preclinical testing protocol which combines the dog model of experimental OA induced by cranial cruciate ligament transection and the Dog model of naturally occurring OA offers the opportunity to further investigate the structural and functional benefits of disease-modifying strategies. Ultimately, a better prediction of outcomes for human clinical trials would be brought.

Regulation of alcohol oxidase 1 (AOX1) promoter and peroxisome biogenesis in different fermentation processes in Pichia pastoris.

Production of recombinant proteins is affected by process conditions, where transcriptional regulation of Pichia pastoris alcohol oxidase 1 (PpAOX1) promoter has been a key factor to influence expression levels of proteins of interest. Here, we demonstrate that the AOX1 promoter and peroxisome biogenesis are regulated based on different process conditions. Two types of GFP-fusion proteins, Ub-R-GFP (short-lived GFP in the cytosol) and GFP-SKL (peroxisomal targeting GFP), were successfully used to characterize the time-course of the AOX1 promoter and peroxisome biogenesis, respectively. The activity of the AOX1 promoter and peroxisome biogenesis was highly subjected to different fermentation process conditions - methanol-limited condition at normoxy (ML), switched feeding of carbon sources (e.g., glucose and methanol) under carbon-limited condition at normoxy (SML), and oxygen-limited (OL) condition. The AOX1 promoter was most active under the ML, but less active under the OL. Peroxisome biogenesis showed a high dependency on methanol consumption. In addition, the proliferation of peroxisomes was inhibited in a medium containing glucose and stimulated in the methanol phase under a carbon-limited fed-batch culture condition. The specific productivity of a monoclonal antibody (qp) under the AOX1 promoter was higher at 86h of induction in the ML than in the OL (0.026 vs 0.020mgg(-1)h(-1)). However, the oxygen-limited condition was a robust process suitable for longer induction (180h) due to high cell fitness. Our study suggests that the maximal production of a recombinant protein is highly dependent on methanol consumption rate that is affected by the availability of methanol and oxygen molecules.

Interobserver agreement and diagnostic accuracy of brain magnetic resonance imaging in dogs.

OBJECTIVE: To evaluate interobserver agreement and diagnostic accuracy of brain MRI in dogs. DESIGN: Evaluation study. ANIMALS: 44 dogs. PROCEDURES: 5 board-certified veterinary radiologists with variable MRI experience interpreted transverse T2-weighted (T2w), T2w fluid-attenuated inversion recovery (FLAIR), and T1-weighted-FLAIR; transverse, sagittal, and dorsal T2w; and T1-weighted-FLAIR postcontrast brain sequences (1.5 T). Several imaging parameters were scored, including the following: lesion (present or absent), lesion characteristics (axial localization, mass effect, edema, hemorrhage, and cavitation), contrast enhancement characteristics, and most likely diagnosis (normal, neoplastic, inflammatory, vascular, metabolic or toxic, or other). Magnetic resonance imaging diagnoses were determined initially without patient information and then repeated, providing history and signalment. For all cases and readers, MRI diagnoses were compared with final diagnoses established with results from histologic examination (when available) or with other pertinent clinical data (CSF analysis, clinical response to treatment, or MRI follow-up). Magnetic resonance scores were compared between examiners with κ statistics. RESULTS: Reading agreement was substantial to almost perfect (0.64 < κ < 0.86) when identifying a brain lesion on MRI; fair to moderate (0.14 < κ < 0.60) when interpreting hemorrhage, edema, and pattern of contrast enhancement; fair to substantial (0.22 < κ < 0.74) for dural tail sign and categorization of margins of enhancement; and moderate to substantial (0.40 < κ < 0.78) for axial localization, presence of mass effect, cavitation, intensity, and distribution of enhancement. Interobserver agreement was moderate to substantial for categories of diagnosis (0.56 < κ < 0.69), and agreement with the final diagnosis was substantial regardless of whether patient information was (0.65 < κ < 0.76) or was not (0.65 < κ < 0.68) provided. CONCLUSIONS AND CLINICAL RELEVANCE: The present study found that whereas some MRI features such as edema and hemorrhage were interpreted less consistently, radiologists were reasonably constant and accurate when providing diagnoses.

Improvement of N-glycan site occupancy of therapeutic glycoproteins produced in Pichia pastoris.

Yeast is capable of performing posttranslational modifications, such as N- or O-glycosylation. It has been demonstrated that N-glycans play critical biological roles in therapeutic glycoproteins by modulating pharmacokinetics and pharmacodynamics. However, N-glycan sites on recombinant glycoproteins produced in yeast can be underglycosylated, and hence, not completely occupied. Genomic homology analysis indicates that the Pichia pastoris oligosaccharyltransferase (OST) complex consists of multiple subunits, including OST1, OST2, OST3, OST4, OST5, OST6, STT3, SWP1, and WBP1. Monoclonal antibodies produced in P. pastoris show that N-glycan site occupancy ranges from 75-85 % and is affected mainly by the OST function, and in part, by process conditions. In this study, we demonstrate that N-glycan site occupancy of antibodies can be improved to greater than 99 %, comparable to that of antibodies produced in mammalian cells (CHO), by overexpressing Leishmania major STT3D (LmSTT3D) under the control of an inducible alcohol oxidase 1 (AOX1) promoter. N-glycan site occupancy of non-antibody glycoproteins such as recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) was also significantly improved, suggesting that LmSTT3D has broad substrate specificity. These results suggest that the glycosylation status of recombinant proteins can be improved by heterologous STT3 expression, which will allow for the customization of therapeutic protein profiles.

A comparison of computed tomography, computed radiography, and film-screen radiography for the detection of canine pulmonary nodules.

Computed tomography (CT) has become more widely available and computed radiography (CR) has replaced film-screen radiography for canine thoracic imaging in many veterinary practices. There are limited data comparing these modalities in a veterinary clinical setting to detect pulmonary nodules. We compared CT, CR, and film-screen radiography for detecting the presence, number, and characteristics of pulmonary nodules in dogs. Observer performance for a variety of experience levels was also evaluated. Twenty-one client-owned dogs with a primary neoplastic process underwent CT and CR; nine also received film-screen radiographs. Positive/negative classification by consensus agreed between the three modalities in 8/9 dogs and between CR and CT in the remaining 12. CT detected the greatest (P = 0.002) total number of nodules and no difference was seen between CR and films. The greatest number of nodules was seen in the right middle and both caudal regions, but only using CT (P < 0.0001). Significantly smaller nodules were detected with CT (P = 0.0007) and no difference in minimum size was detected between CR and films. Observer accuracy was high for all modalities; particularly for CT (90.5-100%) and for the senior radiologist (90.5-100%). CT was also characterized by the least interobserver variability. Although CT, CR, and film-screen performed similarly in determining the presence or absence of pulmonary nodules, a greater number of smaller nodules was detected with CT, and CT was associated with greater diagnostic confidence and observer accuracy and agreement.

Evaluation of osteoarthritis in cats: novel information from a pilot study.

OBJECTIVE: To describe structural changes associated with osteoarthritis (OA) in cats and to quantify OA-associated disability using functional evaluations. STUDY DESIGN: Cross-sectional pilot study with longitudinal data. ANIMALS: Normal cats (n = 2) and coxofemoral joint OA cats (n = 4) were evaluated by physical examination, radiography, and magnetic resonance imaging (MRI). METHODS: Structural changes related to OA were scored using computed radiographs (CR) and MRI. Functional evaluation consisted of podobarometric gait analyses performed using a pressure-sensitive mattress and motor activity assessments using collar-attached, accelerometer-based activity sensors. RESULTS: Structural scores for the coxofemoral joint OA-related lesions were lower in normal cats than OA cats for MRI (P = .07). Use of MRI allowed for whole-organ assessment of the coxofemoral joint. Pelvic limb peak vertical ground reaction force (PVF) was higher in normal cats than OA cats (P = .10). During the night, motor activity was greater in normal cats than OA cats (P = .04). PVF was positively correlated with mean motor activity (Spearman coefficient [Rho] = 0.83, P = .04) and negatively correlated with age and MRI structural score (Rho = -0.93 and -0.79, P < .01 and .06, respectively). CONCLUSIONS: This study provides the first description of OA-related lesions in cats using MRI. Gait analysis and accelerometry should be considered as objective tools to characterize OA-associated disability, although these assessments were weakly correlated with structural changes.

Effect of acquisition time and chemical fat suppression on meningeal enhancement on MR imaging in dogs.

Our purpose was to characterize meningeal gadolinium enhancement on magnetic resonance (MR) imaging in dogs with inflammatory and neoplastic diseases, and to assess interobserver variability and the impact of delayed acquisition and chemical fat saturation on its conspicuity. Transverse T1-weighted FLAIR images were acquired prior to, and immediately following gadolinium injection (T0), and at 5 (T5) and 15-20 min delay (TD), with and without fat suppression, in 155 consecutive dogs imaged for suspected brain disease. The agreement on meningeal enhancement was globally substantial (kappa = 0.61) and the likelihood of obtaining a definite diagnosis was significantly increased with the use of fat suppression (P < or = 0.004). Meningeal enhancement was judged definitively present by consensus in 46 of 155 (30%) dogs. Of these, meningeal enhancement was characterized qualitatively and quantitatively in 30 dogs with a clinical diagnosis (18 inflammatory, 11 neoplastic, 1 infarct), and image sequences were compared. Meningeal enhancement was more often diffuse and leptomeningeal in animals with inflammation versus neoplasia (50% vs. 42%, and 69% vs. 48%, respectively), but significant associations were not found. Meningeal thickness and contrast ratio were higher with neoplasia (P < or = 0.02), but results did not vary significantly between series for either group. Yet, images with fat suppression were most useful 50% of the time for definite diagnosis and/or characterization of meningeal enhancement. While delayed image acquisition following gadolinium injection does not improve characterization of meningeal enhancement in dogs, fat suppression is beneficial qualitatively.

Optimization of erythropoietin production with controlled glycosylation-PEGylated erythropoietin produced in glycoengineered Pichia pastoris.

Pichia pastoris is a methylotropic yeast that has gained great importance as an organism for protein expression in recent years. Here, we report the expression of recombinant human erythropoietin (rhEPO) in glycoengineered P. pastoris. We show that glycosylation fidelity is maintained in fermentation volumes spanning six orders of magnitude and that the protein can be purified to high homogeneity. In order to increase the half-life of rhEPO, the purified protein was coupled to polyethylene glycol (PEG) and then compared to the currently marketed erythropoiesis stimulating agent, Aranesp(®) (darbepoetin). In in vitro cell proliferation assays the PEGylated protein was slightly, and the non-PEGylated protein was significantly more active than comparator. Pharmacodynamics as well as pharmacokinetic activity of PEGylated rhEPO in animals was comparable to that of Aranesp(®). Taken together, our results show that glycoengineered P. pastoris is a suitable production host for rhEPO, yielding an active biologic that is comparable to those produced in current mammalian host systems.

Palpation and dorsal acetabular rim radiographic projection for early detection of canine hip dysplasia: a prospective study.

OBJECTIVE: To determine the value of 2 diagnostic methods: (1) the reduction angle (RA) using the Ortolani maneuver and (2) the dorsal acetabular slope (DAS) from the dorsal acetabular rim (DAR) radiographic projection, to predict osteoarthritis (OA) in dogs with hip dysplasia. STUDY DESIGN: Prospective study. SAMPLE POPULATION: Dogs (n = 73). METHODS: Hip-extended ventrodorsal (VD) radiographic projections, RA, and DAS were evaluated when dogs were 6, 12, and 24 months of age. VD projections were qualitatively scored for OA. RA was determined using the Ortolani maneuver in dorsal recumbency and DAS using the DAR projection. Distraction index (DI) measurements from the compression-distraction radiographic projections (PennHIP method) were also performed at 6 months of age. Statistical analyses were used to establish the range of values of normal and abnormal RA and DAS, to document the temporal variation in RA and DAS, to compare the ability of the different methods to predict coxofemoral OA, to determine the influence of pure passive laxity and of the DAS on the occurrence of an Ortolani sign and on the magnitude of the RA, to establish the relationship between the DAS and subsequent development of passive laxity and coxofemoral OA, and to evaluate the influence of the DAS and RA on the occurrence of coxofemoral OA with severe, moderate, and minimal coxofemoral passive joint laxity, respectively. RESULTS: VD, RA, DAS, and DI methods of coxofemoral joint evaluation correlated significantly with the status of the coxofemoral joints at 2 years of age. The risk of occurrence of coxofemoral OA increased, as the RA, DAS, or DI increased. There was a significant positive linear relationship between RA and DI (P = .015, r(2) = 0.32), RA and DAS (P = .0078, r(2) = 0.38), and DAS and DI (P = .015, r(2) = 0.33). A negative Ortolani sign was at all times significantly predictive of absence of coxofemoral OA at 2 years of age. DAS best predicted coxofemoral OA for DI ≥ 0.7, whereas RA best predicted coxofemoral OA for 0.3 < DI < 0.7; however, RA proved to be the best overall predictor of coxofemoral OA. CONCLUSION: RA measured at 6 months of age in dorsal recumbency was the best predictor of coxofemoral OA at 2 years of age.

Ultrasound-guided percutaneous antegrade pyelography with computed tomography for the diagnosis of spontaneous partial ureteral rupture in a dog.

A 10-year-old spayed female dalmatian dog developed acute vomiting and abdominal pain. Ultrasound examination of the abdomen showed right hydronephrosis and proximal ureter dilation with mild retroperitoneal free fluid. Computed tomography (CT) of the abdomen confirmed the ultrasonographic findings and revealed, additionally, a right ureteral stone. Spontaneus rupture of the right ureter was confirmed with CT post ultrasound-guided percutaneous antegrade pyelography. Pyeloureteral rupture and the presence of a ureteral stone were confirmed at surgery.

Pyélographie antégrade percutanée guidée par échographie avec tomodensitométrie pour le diagnostic d'une rupture urétrale partielle spontanée chez un chien. Une chienne Dalmatien stérilisée âgée de 10 ans a manifesté des vomissements et de la douleur abdominale aigus. Une échographie de l'abdomen a montré de l'hydronéphrose à droite et une dilatation proximale de l'urètre avec un peu de liquide rétropéritonéal libre. Une tomodensitométrie de l'abdomen a confirmé les résultats de l'échographie et a révélé, en plus, un calcul urétéral droit. Une rupture spontanée de l'urètre droit a été confirmée par tomodensitométrie après une pyélographie antégrade percutanée guidée par échographie. La rupture pyélo-urétérale et la présence de calcul urétéral ont été confirmées à la chirurgie.(Traduit par Isabelle Vallières).

Optimization of a glycoengineered Pichia pastoris cultivation process for commercial antibody production.

Glycoengineering enabled the production of proteins with human N-linked glycans by Pichia pastoris. This study used a glycoengineered P. pastoris strain which is capable of producing humanized glycoprotein with terminal galactose for monoclonal antibody production. A design of experiments approach was used to optimize the process parameters. Followed by further optimization of the specific methanol feed rate, induction duration, and the initial induction biomass, the resulting process yielded up to 1.6 g/L of monoclonal antibody. This process was also scaled-up to 1,200-L scale, and the process profiles, productivity, and product quality were comparable with 30-L scale. The successful scale-up demonstrated that this glycoengineered P. pastoris fermentation process is a robust and commercially viable process.