Predicting likelihood of in vivo chemotherapy response in canine lymphoma using ex vivo drug sensitivity and immunophenotyping data in a machine learning model

We report a precision medicine platform that evaluates the probability of chemotherapy drug efficacy for canine lymphoma by combining ex vivo chemosensitivity and immunophenotyping assays with computational modelling. We isolated live cancer cells from fresh fine needle aspirates of affected lymph nodes and collected post‐treatment clinical responses in 261 canine lymphoma patients scheduled to receive at least 1 of 5 common chemotherapy agents (doxorubicin, vincristine, cyclophosphamide, lomustine and rabacfosadine). We used flow cytometry analysis for immunophenotyping and ex vivo chemosensitivity testing. For each drug, 70% of treated patients were randomly selected to train a random forest model to predict the probability of positive Veterinary Cooperative Oncology Group (VCOG) clinical response based on input variables including antigen expression profiles and treatment sensitivity readouts for each patient's cancer cells. The remaining 30% of patients were used to test model performance. Most models showed a test set ROC‐AUC > 0.65, and all models had overall ROC‐AUC > 0.95. Predicted response scores significantly distinguished (P < .001) positive responses from negative responses in B‐cell and T‐cell disease and newly diagnosed and relapsed patients. Patient groups with predicted response scores >50% showed a statistically significant reduction (log‐rank P < .05) in time to complete response when compared to the groups with scores <50%. The computational models developed in this study enabled the conversion of ex vivo cell‐based chemosensitivity assay results into a predicted probability of in vivo therapeutic efficacy, which may help improve treatment outcomes of individual canine lymphoma patients by providing predictive estimates of positive treatment response.     

Limb‐sparing in dogs using patient‐specific,three‐dimensional‐printed endoprosthesis for distal radial osteosarcoma: A pilot study

Limb‐sparing for distal radial osteosarcoma has a high rate of complications. Using personalized three‐dimensional (3D)‐printed implants might improve outcome. The goals of this study were to optimize use of patient‐specific, 3D‐printed endoprostheses for limb‐sparing in dogs in the clinical environment and to report the outcome. This was a pilot study where five client‐owned dogs were enrolled. Computed tomography (CT) of the thoracic limbs was performed, which was used to create patient‐specific endoprostheses and cutting guides, and repeated on the day of surgery. Intra‐arterial (IA) carboplatin was introduced in the clinical management. Limb‐sparing was performed. Outcome measures were time required to produce the endoprosthesis and cutting guide, fit between cutting guide and endoprosthesis with host bones, gait analysis, size of the tumour, percent tumour necrosis, complications, disease‐free interval (DFI) and survival time (ST). Four dogs received IA carboplatin. Excessive tumour growth between planning CT and surgery did not occur in any dog. The interval between the CT and surgery ranged from 14 to 70 days. Fit between the cutting‐guide and endoprosthesis with the host bones was good to excellent. At least one complication occurred in all dogs. Two dogs were euthanized with STs of 192 and 531 days. The other dogs were alive with a follow up of 534 to 575 days. IA chemotherapy is a promising strategy to minimize the risk of excessive tumour growth while waiting for the endoprosthesis and cutting‐guide to be made. The design of the cutting‐guide was critical for best fit of the endoprosthesis with host bones.     

Disseminated fungal infection in two species of captive sharks

In this report, two cases of systemic mycosis in captive sharks are characterized. These cases were progressive and ultimately culminated in terminal disease. Paecilomyces lilacinus, an uncommon pathogen in human and veterinary medicine, was associated with areas of necrosis in the liver, heart, and gill in a great hammerhead shark (Sphyrna mokarran). Fungal growth was observed from samples of kidney, spleen, spinal fluid, and coelomic cavity swabs. Dual fungal infection by Exophiala pisciphila and Mucor circinelloides was diagnosed in a juvenile zebra shark (Stegostoma fasciatum). Both fungi were present in the liver, with more severe tissue destruction associated with E. pisciphila. E. pisciphila also produced significant necrosis in the spleen and gill, while M. circinelloides was associated with only minimal tissue changes in the heart. Fungal cultures from liver, kidney, and spleen were positive for both E. pisciphila and M. circinelloides. Identification of P. lilacinus and M. circinelloides was based on colonial and hyphal morphology. E. pisciphila was identified by sequence analysis of the 28S rRNA D1/D2 region and the internal transcribed spacer (ITS) region between the 18S and 28S rRNA subunit. These cases, and a lack of information in the literature, highlight the need for further research and diagnostic sampling to further characterize the host-pathogen interaction between elasmobranchs and fungi.     

Hard White Versus Hard Red Wheats: Taste Tests and Milling and Baking Properties

Molecular markers for the red grain color (R) loci controlling seed color and the polyphenol oxidase (Ppo‐A1) locus controlling polyphenol oxidase (PPO) activity in seed have recently been developed. These markers provided the opportunity to convert the hard red spring wheat cultivars Choteau and Hank to white‐seeded versions with high and low PPO levels, respectively. These sets of near‐isogenic lines provided material to test the effects of seed color and PPO activity on a range of end‐use quality traits. We tested recurrent parents Choteau and Hank, along with near‐isogenic derivatives with white seed, in two replicated trials in Bozeman, Montana, for end‐use quality parameters. The white‐seeded lines consisted of both high‐ and low‐PPO near‐isogenic lines. The primary impact of white seed was the production of whole wheat bread with a perceived sweeter taste relative to the red‐seeded lines. Noodle color was not consistently impacted by the level of PPO variation despite relatively large reductions in PPO level. The alleles for white seed color did not appear to impact agronomic traits. These results suggested that hard white low‐PPO hard spring wheat would be advantageous in terms of conferring brighter color to Asian noodles and a sweeter taste to whole wheat bread.     

Response of sorghum stalk pathogens to <Emphasis Type="Italic">brown midrib</Emphasis> plants and soluble phenolic extracts from near isogenic lines

Sorghum [Sorghum bicolor (L.) Moench] has drawn attention as potential feedstock for lignocellulosic biofuels production, and reducing lignin is one way to increase conversion efficiency. Little research has been previously conducted to assess the response of reduced lignin sorghum lines to the Fusarium stalk rot pathogens Fusarium verticillioides and Fusarium proliferatum and the charcoal rot pathogen, Macrophomina phaseolina. Loss of function mutations in either the Brown midrib (Bmr) 6 or 12 gene that both encode a monolignol biosynthetic enzyme in the pathway that produces subunits of the lignin polymer, results in reduced lignin content. Near-isogenic bmr6, bmr12, and bmr6 bmr12 lines had previously been developed, which were shown to have significantly reduced lignin content and increased levels of soluble phenolics. In the current study, these lines in two backgrounds were shown to not be more susceptible to F. verticillioides, F. proliferatum and M. phaseolina inoculations, and some bmr lines exhibited increased resistance to F. proliferatum and M. phaseolina, compared to wild-type lines. When the Fusarium stalk rot pathogen, Fusarium thapsinum, was grown on methanol soluble stalk extracts from bmr6 and wild-type plants, it grew significantly faster on medium with bmr6 extract than on wild-type extract or controls. This result suggested that factors other than soluble phenolics from the extract, such as cell wall bound phenolics or inducible defense compounds, contributed to increased resistance observed in bmr6 plants.     

Semilobar holoprosencephaly in a Morgan horse

A6 1/2‐month‐old Morgan filly was examined because of a history of abnormal behavior, teeth grinding, hypothermia, and electrolyte disturbances when weaned. She was from a breeding farm with several other Morgan horses. The 11‐year‐old dam had been purchased the year before as a proven broodmare, which had several previous foals. Breeding, gestation, and birth of this foal were normal. She was raised with 4 other mares and their offspring on pasture with free access to shelter in an open barn. Supplementary feeding consisted of oats and timothy hay. The owners reported that the foal showed unusual behavior, such as lack of apprehension of people, lack of distress from maternal separation, and a higher activity level than other foals of the same age. The foal extensively chewed the dam's tail and mane, masticated oats slowly with rapid jaw movements without actually swallowing them, and ground her teeth. She frequently nibbled the handler's clothes without biting, ate pebbles, and played with the salt block in the paddock. At 4 1/2 months of age, she was treated for suspected gastroduodenal ulcers and weaned. The referring veterinarian examined her 5 days after weaning because of dull demeanor and excessive teeth grinding. The foal was in thin body condition, hypothermic (37°C, 98.6°F), and tachycardic (60 beats per minute [bpm]) and had decreased borborygmi. Major abnormalities on serum biochemistry were severe hypernatremia (166 mmol/L; reference range 136–144 mmol/L) and hyperchloremia (128 mmol/L; reference range 94–104 mmol/L), azotemia (urea, 11.3 mmol/L; reference range 4.2–8.9 mmol/L), and hyperfibrinogenemia (5.2 g/L, reference range 1.6–2.9 g/L). The only abnormality on the CBC was hemoconcentration (PCV, 0.57 L/L; reference range 0.28–0.44 L/L). The foal was treated with penicillin procaine G^a (20,000 IU/kg [9072 IU/1b] IM q12h) and rifampin^b (5 mg/kg [2.7 mg/1b] PO q8h). The next day the tachycardia worsened (120 bpm) and the foal was estimated to be 5–8% dehydrated. IV fluid therapy with lactated Ringer solution^c (LRS) was initiated, and the antibiotic was changed to ceftiofur^d (2 mg/kg [0.91 mg/1b] IV q12h). The foal and dam were rejoined, and the foal's clinical status improved with resumption of nursing. Serial laboratory testing showed persistent hypernatremia 160 mmol/L) and hyperchloremia (123 mmol/L), azotemia urea 11.3 mmol/L and creatinine 168 umol/L; reference range 80–130 μmol/L), hyperglycemia (8.7 mmol/L; reference range 3.7–6.7 mmol/L), high aspartate aminotranferase activity (662 U/L; reference range 259–595 U/L), and high creatine kinase (CK) activity (1,196 U/L; reference range 108–430 U/L). The foal's condition improved and IV fluids were discontinued. Ceftiofur administration was discontinued and trimethoprim‐sulfamethoxazole^e (25 mg/kg [11.3 mg/1b] PO q12h) was administered for 3 days. During the next month the foal was stable but the abnormal behavior persisted. She was weaned again, and within days marked behavior changes such as circling, throwing the head around compulsively, and severe hind‐end shivering recurred. At examination, the foal was dull, tachycardic (60 bpm), was hypothermic (33.6°C, 92.5°F), had dark red mucous membranes, and was estimated to be 5% dehydrated. Laboratory findings were similar to those of the previous tests except for high fibrinogen (7.1 g/L). The foal was again rejoined with the dam, treated with intramuscular penicillin, and referred     

Hemophagocytosis and Histoplasma‐like fungal infection in 32 cats

Histoplasmosis is one of the most common systemic fungal diseases in cats from the United States. It commonly causes respiratory or disseminated disease and is often associated with one or more cytopenias. Here, we describe 32 cats in which a Histoplasma‐like fungal infection was associated with concurrent hemophagia in at least one sample site, commonly spleen, bone marrow, liver, and/or lymph node. The degree of hemophagia was characterized as moderate or marked in the majority of cases, and in all cases, there was a predominance of phagocytized mature erythrocytes. A few cases also had macrophages with phagocytized erythroid precursors, platelets, and/or neutrophils. Complete blood count results were available for 25 cats, and cytopenias were common (20/25), including solitary anemia (10), anemia and thrombocytopenia (5), solitary neutropenia (2), pancytopenia (2), and anemia and neutropenia (1). Bone marrow samples were only available in a small subset of cases, preventing the further assessment of the causes of the cytopenias. Hemophagocytosis has been previously reported in cats with neoplastic diseases and a cat with calicivirus infection, and likely occurs with other conditions as well, such as hemorrhage or hemolysis. Results of this report suggest that systemic fungal disease is an additional differential to consider when there is hemophagia in a feline cytology sample.