Successful drug development despite adverse preclinical findings part 1: processes to address issues and most important findings

Unexpected adverse preclinical findings (APFs) are not infrequently encountered during drug development. Such APFs can be functional disturbances such as QT prolongation, morphological toxicity or carcinogenicity. The latter is of particular concern in conjunction with equivocal genotoxicity results. The toxicologic pathologist plays an important role in recognizing these effects, in helping to characterize them, to evaluate their risk for man, and in proposing measures to mitigate the risk particularly in early clinical trials. A careful scientific evaluation is crucial while termination of the development of a potentially useful drug must be avoided. This first part of the review discusses processes to address unexpected APFs and provides an overview over typical APFs in particular classes of drugs. If the mode of action (MoA) by which a drug candidate produces an APF is known, this supports evaluation of its relevance for humans. Tailor-made mechanistic studies, when needed, must be planned carefully to test one or several hypotheses regarding the potential MoA and to provide further data for risk evaluation. Safety considerations are based on exposure at no-observed-adverse-effect levels (NOAEL) of the most sensitive and relevant animal species and guide dose escalation in clinical trials. The availability of early markers of toxicity for monitoring of humans adds further safety to clinical studies. Risk evaluation is concluded by a weight of evidence analysis (WoE) with an array of parameters including drug use, medical need and alternatives on the market. In the second part of this review relevant examples of APFs will be discussed in more detail.     

Seroprevalence of bovine viral diarrhea virus neutralizing antibodies in finisher hogs in Ontario swine herds and targeted diagnostic testing of 2 suspect herds

A pilot study was initiated to determine the seroprevalence of bovine viral diarrhea virus (BVDV) neutralizing antibodies in finisher hogs in Ontario swine herds, including 2 swine herds with clinical syndromes suspicious of BVDV. No herds were positive for BVDV antibodies by virus neutralization. The 2 swine herds with clinical disease suggestive of pestivirus infection were also negative for antibodies to BVDV in indirect fluorescent antibody assays. Prevalence of BVDV in Ontario swine farms is negligible.     

Artifacts in digital radiography

Digital radiography is becoming more prevalent in veterinary medicine, and with its increased use has come the recognition of a number of artifacts. Artifacts in digital radiography can decrease image quality and mask or mimic pathologic changes. They can be categorized according to the step during which they are created and include preexposure, exposure, postexposure, reading, and workstation artifacts. The recognition and understanding of artifacts in digital radiography facilitates their reduction and decreases misinterpretation. The purpose of this review is to name, describe the appearance, identify the cause, and provide methods of resolution of artifacts in digital radiography.     

Effects of forage type,body condition and single‐nucleotide polymorphisms in the bovine cytochrome P450 regulatory region on cow productivity*

Single‐nucleotide polymorphisms (SNP) in the coding sequence of cytochrome p450 (CYP3A28) have been associated with milk yield and composition, and calving traits in cows. In this study, we aimed to determine whether (i) the CYP3A28 regulatory region was polymorphic and (ii) SNP genotype, forage type, body condition and their interactions affect cow productivity. Primers for CYP3A28 promoter were designed to amplify a 483‐bp segment by PCR. Amplicon sequences revealed seven SNP (T‐318C, T‐113A, C‐189T, T‐78G, A6G, G17A and T21C) in Brahman (38 cows), Brahman x Angus reciprocal crosses (47 cows) and crossbreds (98 cows). Angus cows (n = 41) appeared to be fixed at those SNP locations. Genotype and forage {endophyte‐infected tall fescue [KY+; Lolium arundinaceum (Schreb.) S. J. Darbyshire] vs. bermudagrass [Cynodon dactylon (L.) Pers.]} effects on lifetime (8‐years) calving rate, and calf weaning weights and heights were determined in Herd 1 (126 cows); genotype and BC (low vs. moderate) effects on calving date and calving percent were determined in Herd 2 (98 cows). Four SNP (T‐318C, T‐113A, A06G and T21C) appeared to be related to cattle productivity, CC cows at T‐318C having a lower (p < 0.05) lifetime calving rate than TC or TT cows (65%, 85% and 81% respectively). Cows that grazed KY+ and were TT at T‐318C produced calves that tended (p < 0.07) to weigh less than their contemporaries. Moreover, calves of TT cows were shorter (p < 0.05) at weaning than calves of CC or TC cows. In Herd 2, moderate‐BC cows that were TT or AA at T‐318C, T‐113A, T‐78G, A6G and T21C had greater (p < 0.05) calving rates (74–80%) than heterozygous cows (46–60%), and low‐BC cows that were AA at G17A calved at least 6 days earlier (p < 0.05) than heterozygous cows. Our findings suggest that SNP in the CYP3A28 regulatory region of Brahman‐influenced cows are associated with cattle productivity.     

Evaluation of an arthroscopic approach for transection of the equine collateral sesamoidean ligament

Objective: To evaluate: (1) an arthroscopic technique for transection of the collateral sesamoidean ligament (CSL); and (2) the healing response using magnetic resonance (MR) and microscopic examination. Study Design: Experimental study. Animals: Adult horses (n=6). Methods: Six sound horses with normal front foot radiographic and MR examinations were used. Lameness examination was performed before surgery and monthly for 12 months. Front foot radiography was performed at 180 and 360 days after surgery. Front foot MR was performed before, and at 7, 90, 180, and 360 days after surgery. Arthroscopic CSL desmotomy was performed on 1 forelimb. Gross and microscopic examination was performed on the CSL from both forelimbs at 360 days after surgery. Lameness scores were compared over time using the nonparametric Friedman's test for paired groups. CSL measurements were compared using paired t‐tests with a 2‐tailed significance level of P<.05. Results: Radiographs remained normal throughout study period. Surgery resulted in lameness on the operated limb for up to 2 months, after which all horses returned to soundness. CSL transection was confirmed during arthroscopy and with MR examination 7 days after surgery. Gross and microscopic evaluation confirmed ligament healing. Conclusions: CSL desmotomy resulted in short‐term lameness after surgery followed by healing of the CSL confirmed by gross and microscopic analysis.     

Suspected Tension Pneumomediastinum In A Dog: A Case Report

Successful resuscitation of a trauma-induced, suspected tension pneumomediastinum in a dog is presented. The cause was a caudal cervical penetration wound which acted as a one-way valve allowing air to enter, but not exit, the cranial mediastium. Early in the course of hospitalization, respiratory and cardiovascular collapse occurred; an emergent parasternotomy allowed decompression of the mediastinum, and the cradiovascular system immediately responded. The dog made a complete recovery following surgical exploration and removal of contaminated mediastinal tissue. Nine days of intensive care included 24-hour monitoring, oxygen therapy, intravenous crystalloids and colloids, continuous pleural suctioning, nutritional support, antibiotics, and pain control.     

Evaluation of pacing site in dogs with naturally occurring complete heart block

Objective

To compare left ventricular synchronization and systolic performance with transvenous pacing of the right ventricular apex (RVA), left ventricular free wall (LVF) or simultaneous pacing of the RVA and LVF (BiV).

Animals, materials and methods

Seven canine patients with complete heart block. Prospective study evaluating effect of pacing site. Twenty four hours following implantation of transvenous BiV pacing systems, electrocardiograms and echocardiograms were assessed during pacing from the: (1) Right Atrial Appendage/RVA (RAA/RVA), (2) RAA/LVF, and (3) RAA/BiV.

Results

QRS duration was significantly shorter with BiV pacing versus LVF pacing (p < 0.001), or RVA pacing (p < 0.001). Echocardiographic indices of systolic performance fractional shortening (FS), ejection fraction (EF), cardiac output (CO) were significantly higher with BiV pacing than with pacing from the RVA (P = 0.023, 0.006, and 0.002 respectively). Cardiac output, measured by the biplane Simpson's method, was higher with LVF versus RVA pacing (P = 0.036). There was no difference in FS or EF when comparing LVF to RVA pacing. Tissue Doppler measurements of synchronization and systolic performance did not show any difference between pacing mode, but a significantly increased number of segments were seen to contract following aortic valve closure during LVF pacing (P = 0.0268) and RVA pacing (P = 0.0197) as compared to BiV pacing.

Conclusions

Findings suggest that BiV pacing improves cardiac output and systolic performance versus RVA pacing. This improvement however, is not reflected in tissue Doppler indices of synchronization and systolic performance.     

Serum complement activity in the three-toed amphiuma (Amphiuma tridactylum)

Some animals routinely endure serious injuries from predators or during intraspecific territorial conflicts. Such is the case for Amphiuma tridactylum, an aquatic salamander that lives in an environment rich in potentially infectious microbes, apparently with rare or no pathogenic infection. Some vertebrates possess innate immune mechanisms, but whether this is the case for Amphiuma is unknown. To assess this potential, plasma from 19 A. tridactylum was pooled and used for characterisation of serum complement activity. The ability of A. tridactylum plasma to hemolyse unsensitised sheep red blood cells (SRBCs) was titer-dependent, with low activity observed even at high plasma titers. The kinetic characterisation of SRBC hemolysis revealed that significant activity could be measured within 10 min of incubation, and maximal activity occurred within 60 min. The SRBC hemolysis by A. tridactylum plasma was also temperature-dependent, with maximal activity at 30 °C. In addition, this activity was sensitive to mild heat treatment, with 96% of activity inhibited by incubation at 56 °C for 30 min. The SRBC hemolysis could also be inactivated by pretreatment of the plasma with proteases, indicating that this activity was protein dependent. The activity required divalent metals ions, with activity inhibited by EDTA, citrate, or phosphate. However, the chelator-inhibited activity could be restored by the addition of excess Ca²⁺ or Mg²⁺, but not Cu²⁺ or Ba²⁺, indicating specificity of the divalent metal ion requirement. The sensitivity to heat, proteases, and divalent metal ion chelators strongly suggests that A. tridactylum plasma-mediated hemolysis of SRBCs is mediated by the serum complement system of proteins.