Mycobacterium genavense granulomatous typhlocolitis in a horse.

A 23-y-old gelding was presented to a veterinary teaching hospital with a history of chronic, refractory diarrhea. Clinically, the horse was in poor body condition, with a thickened and corrugated large intestine identified by transcutaneous abdominal ultrasonography. At postmortem examination following euthanasia, the large colon and cecum had segmental thickening of the intestinal wall with innumerable mucosal ulcers and prominent polypoid mucosal masses. Many mesenteric and hepatic lymph nodes were enlarged. Histology revealed granulomatous and ulcerative typhlocolitis and granulomatous lymphadenitis with myriad acid-fast, variably gram-positive, intrahistiocytic bacilli that stained by immunohistochemistry for mycobacteria. Molecular testing by PCR and sequencing identified the causative agent as Mycobacterium genavense, which is an unusual presentation of infection in a horse.

Intrahepatic mucinous cholangiocarcinoma with recurrent colic in a horse: case report and literature review of cholangiocarcinoma in horses.

A 17-y-old Arabian mare was presented to the Auburn Large Animal Veterinary Teaching Hospital with a long-term history of intermittent mild recurrent colic that responded to medical treatment. CBC revealed mild lymphopenia; serum biochemistry findings were of increased gamma-glutamyl transferase and creatine kinase activities, hyperferremia, hyperglycemia, hypomagnesemia, and hypokalemia. Abdominocentesis was compatible with low-protein transudate. Due to the progression and duration of clinical signs, the owner elected euthanasia. Postmortem examination and histopathology confirmed a cholangiocarcinoma. The neoplastic cells were arranged in large cysts containing lakes of mucin that comprised 90% of the tumor volume; thus, a mucinous variant was determined. The neoplastic cells had strong cytoplasmic immunolabeling for cytokeratin 19 and lacked immunolabeling for hepatocyte paraffin 1, supporting bile duct origin. Cholangiocarcinomas are infrequent tumors in horses with nonspecific and slow progressive clinical signs, including recurrent colic. Mucinous cholangiocarcinomas are seldom reported in veterinary medicine and, to our knowledge, have not been reported previously in horses.

Prognostic value of histopathologic traits independent of stromal tumor-infiltrating lymphocyte levels in chemotherapy-naïve patients with triple-negative breast cancer.

BACKGROUND: In the absence of prognostic biomarkers, most patients with early-stage triple-negative breast cancer (eTNBC) are treated with combination chemotherapy. The identification of biomarkers to select patients for whom treatment de-escalation or escalation could be considered remains an unmet need. We evaluated the prognostic value of histopathologic traits in a unique cohort of young, (neo)adjuvant chemotherapy-naïve patients with early-stage (stage I or II), node-negative TNBC and long-term follow-up, in relation to stromal tumor-infiltrating lymphocytes (sTILs) for which the prognostic value was recently reported. MATERIALS AND METHODS: We studied all 485 patients with node-negative eTNBC from the population-based PARADIGM cohort which selected women aged <40 years diagnosed between 1989 and 2000. None of the patients had received (neo)adjuvant chemotherapy according to standard practice at the time. Associations between histopathologic traits and breast cancer-specific survival (BCSS) were analyzed with Cox proportional hazard models. RESULTS: With a median follow-up of 20.0 years, an independent prognostic value for BCSS was observed for lymphovascular invasion (LVI) [adjusted (adj.) hazard ratio (HR) 2.35, 95% confidence interval (CI) 1.49-3.69], fibrotic focus (adj. HR 1.61, 95% CI 1.09-2.37) and sTILs (per 10% increment adj. HR 0.75, 95% CI 0.69-0.82). In the sTILs <30% subgroup, the presence of LVI resulted in a higher cumulative incidence of breast cancer death (at 20 years, 58%; 95% CI 41% to 72%) compared with when LVI was absent (at 20 years, 32%; 95% CI 26% to 39%). In the ≥75% sTILs subgroup, the presence of LVI might be associated with poor survival (HR 11.45, 95% CI 0.71-182.36, two deaths). We confirm the lack of prognostic value of androgen receptor expression and human epidermal growth factor receptor 2 -low status. CONCLUSIONS: sTILs, LVI and fibrotic focus provide independent prognostic information in young women with node-negative eTNBC. Our results are of importance for the selection of patients for de-escalation and escalation trials.

Doxorubicin and zoledronate treatment in a dog with hemophagocytic histiocytic sarcoma.

A 6-year-old castrated male greyhound dog was referred for hemophagocytic histiocytic sarcoma (HHS) diagnosed following splenectomy. Severe thrombocytopenia, mild hypoalbuminemia, mild hypocholesterolemia, and mild hyperbilirubinemia were present. Abdominal ultrasound findings were concerning for hepatic metastasis. Doxorubicin and zoledronate combination therapy was initiated. The dog improved clinically and its thrombocytopenia, hypoalbuminemia, and hyperbilirubinemia resolved. The dog appeared well for 147 d before tumor progression was noted. The dog was treated with lomustine as a final measure, with no response. The dog survived for 6 mo with chemotherapy. To the authors' knowledge, this is the first report of clinical benefit of chemotherapy for HHS. Key clinical message: Doxorubicin should be considered for treating canine HHS since this variant of the disease is historically refractory to lomustine. Further research regarding efficacy of doxorubicin and zoledronate should be pursued.

Traitement à la doxorubicine et au zolédronate chez un chien atteint de sarcome histiocytaire hémophagocytaire. Un lévrier mâle castré de 6 ans a été vu pour un sarcome histiocytaire hémophagocytaire (HHS) diagnostiqué à la suite d'une splénectomie. Une thrombopénie sévère, une hypoalbuminémie légère, une hypocholestérolémie légère et une hyperbilirubinémie légère étaient présentes. Les résultats de l'échographie abdominale étaient préoccupants quant aux métastases hépatiques. Un traitement associant doxorubicine et zolédronate a été instauré. Le chien s'est amélioré cliniquement et sa thrombocytopénie, son hypoalbuminémie et son hyperbilirubinémie ont disparu. Le chien semblait en bonne santé pendant 147 jours avant de constater une progression tumorale. Le chien a été traité avec de la lomustine comme mesure finale, sans réponse. Le chien a survécu 6 mois grâce à la chimiothérapie. À la connaissance des auteurs, il s'agit du premier rapport faisant état d'un bénéfice clinique de la chimiothérapie pour le HHS.Message clinique clé :La doxorubicine doit être envisagée pour traiter le HHS canin puisque cette variante de la maladie est historiquement réfractaire à la lomustine. Des recherches plus approfondies concernant l'efficacité de la doxorubicine et du zolédronate devraient être poursuivies.(Traduit par Dr Serge Messier).

Enzalutamide Reduces Oxycodone Exposure in Men with Prostate Cancer.

BACKGROUND AND OBJECTIVE: Up to 90% of patients with castration-resistant prostate cancer (CRPC) will develop symptomatic bone metastases requiring pain medication, with opioids being the mainstay of therapy in treating moderate and severe pain. Enzalutamide is an androgen receptor antagonist for the treatment of CRPC and a strong inducer of cytochrome P450 (CYP)3A4. Hereby, enzalutamide potentially reduces the exposure of oxycodone, an opioid metabolized by CYP3A4 and CYP2D6. Our objective was to evaluate the potential drug-drug interaction of enzalutamide and oxycodone. METHODS: A prospective, nonrandomized, open-label, two-arm parallel study was performed. All patients received a single dose of 15 mg normal-release oxycodone. Patients in the enzalutamide arm (ENZ-arm) received enzalutamide 160 mg once daily. Plasma concentrations of oxycodone and its metabolites were quantified using a validated liquid chromatography with tandem mass spectrometry (LC-MS/MS) method. RESULTS: Twenty-six patients (13 ENZ-arm; 13 control arm) were enrolled in the study. Enzalutamide decreased the mean AUC0-8 h and Cmax of oxycodone with, respectively, 44.7% (p < 0.001) and 35.5% (p = 0.004) compared with the control arm. The AUC0-8 h and Cmax of the active metabolite oxymorphone were 74.2% (p < 0.001) and 56.0% (p = 0.001) lower in the ENZ-arm compared with the control arm. In contrast, AUC0-8 h and Cmax of the inactive metabolites noroxycodone and noroxymorphone were significantly increased by enzalutamide. CONCLUSION: Co-administration of enzalutamide significantly reduced exposure to oxycodone and its active metabolite oxymorphone in men with prostate cancer. This should be taken into account when prescribing enzalutamide combined with oxycodone.

Chylothorax due to thrombosis of the jugular and subclavian veins in a patient with gastric cancer: a case report.

BACKGROUND: Chylothorax is a rare condition due to leakage of chyle in the thoracic cavity. When large amounts of chyle leak into the thoracic cavity, it can lead to severe respiratory, immune, and metabolic complications. Chylothorax has many potential underlying etiologies, and the most common causes are traumatic chylothorax and lymphoma. Venous thrombosis of the upper extremities is a rare cause of a chylothorax. CASE PRESENTATION: A 62-year-old Dutch man with a medical history of gastric cancer, treated with neoadjuvant chemotherapy and surgery 13 months prior, presented with dyspnea and a swollen left arm. Computed tomography thorax showed bilateral pleural effusion that was more prominent on the left side. The computed tomography scan further revealed thrombosis of the left jugular and subclavian veins and osseal masses suggesting cancer metastasis. Thoracentesis was performed to confirm the suspicion of gastric cancer metastasis. The obtained fluid was milky with a high level of triglycerides, but contained no malignant cells; hence, the diagnosis of the pleural effusion was chylothorax. Treatment with anticoagulation and a medium-chain-triglycerides diet was started. Furthermore, bone metastasis was confirmed with a bone biopsy. CONCLUSION: Our case report demonstrates chylothorax as a rare cause of dyspnea in a patient with pleural effusion and a history of cancer. Therefore, this diagnosis should be considered in all patients with a history of cancer with new-onset pleural effusion and thrombosis of the upper extremities or clavicular/mediastinal lymphadenopathy.

Classification of middle- and long-distance runners based upon their performance.

The scientific literature lacks consensus on classification of middle- and long-distance runners. This creates situations where the sample studied may not represent the target population and could produce misleading conclusions. Thus, we present an approach for a data-driven classification of middle- and long-distance runners according to their competition results. The best annual results of middle- and long-distance track runners participating at major (Olympics, World and European Championships) and national championships (Denmark, Sweden, Finland, Norway) were gathered for the 2012-2018 period. Overall, 1920 men's and 1808 women's performance results were gathered. The results were grouped accordingly. Quadratic discriminant analysis was applied to define the limits between the groups. Three basic categories could be proposed for classification: world class, international and national. Classification provides a realistic overview of performance standards and the number of athletes for different categories in middle- and long-distance track running in real-world settings. The performance-based classification provides data-driven and unified criteria for reporting standards on athletes' proficiency levels. It allows for more consistent reporting practices on the target population in research. In addition to scientific research, the classification could also be employed for a variety of practical purposes.

Therapeutic drug monitoring-based precision dosing of oral targeted therapies in oncology: a prospective multicenter study.

BACKGROUND: Oral targeted therapies show a high pharmacokinetic (PK) interpatient variability. Even though exposure has been positively correlated with efficacy for many of these drugs, these are still dosed using a one-size-fits-all approach. Consequently, individuals have a high probability to be either underexposed or overexposed, potentially leading to suboptimal outcomes. Therapeutic drug monitoring, which is personalized dosing based on measured systemic drug concentrations, could address these problems. PATIENTS AND METHODS: Patients were enrolled in this prospective multicenter study (www.trialregister.nl; NL6695) if they started treatment with one of the 24 participating oral targeted therapies. Primary outcome was to halve the proportion of underexposed patients, compared with historical data. PK sampling was carried out after 4, 8 and 12 weeks, and every 12 weeks thereafter. In case of Cmin below the predefined target and manageable toxicity, a pharmacokinetically guided intervention was proposed (i.e. checking compliance and drug-drug interactions, concomitant intake with food, splitting intake moments or dose increments). RESULTS: In total, 600 patients were included of whom 426 patients are assessable for the primary outcome and 552 patients had ≥1 PK sample(s) available and were therefore assessable for the overall analyses. Pharmacokinetically guided dosing reduced the proportion of underexposed patients at the third PK measurement by 39.0% (95% confidence interval 28.0% to 49.0%) compared with historical data. At the third PK measurement, 110 out of 426 patients (25.8%) had a low exposure. In total, 294 patients (53.3%) had ≥1 PK sample(s) below the preset target at a certain time point during treatment. In 166 of these patients (56.5%), pharmacokinetically guided interventions were carried out, which were successful in 113 out of 152 assessable patients (74.3%). CONCLUSIONS: Pharmacokinetically guided dose optimization of oral targeted therapies was feasible in clinical practice and reduced the proportion of underexposed patients considerably.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Partial parotid sialoadenectomy in a horse with parotid ductal carcinoma: Surgical description and case report.

OBJECTIVES: To describe the surgical findings, histopathological features, and long-term outcome for a horse with parotid salivary carcinoma. STUDY DESIGN: Case report ANIMALS: Twelve year old American Quarter Horse gelding. METHODS: The gelding was presented for a 10 × 10 cm swelling below the base of the right ear. Ultrasonographic examination revealed a mass involving the right parotid salivary gland. Incisional biopsy was consistent with parotid carcinoma. The tumor was marginally excised. The lateral wall of the guttural pouch was excised with the mass and was reconstructed with a porcine small intestinal submucosal (SIS) sheet. Cisplatin beads were implanted in the wound bed prior to closure. Firocoxib (0.1 mg/kg orally, daily, every 24 h) treatment was initiated. RESULTS: Postoperative complications included right-sided facial nerve paralysis, difficulty with deglutition of fibrous feeds, and surgical site dehiscence. Wound healing was achieved by second intention. Partial improvement in nerve function was observed within the first 6 months. At 12 months postparotidectomy, no sign of tumor reoccurrence or metastatic disease was present, and the gelding returned to work. CONCLUSION: Partial parotid sialoadenectomy was performed with a favorable long-term outcome. Regional anatomic knowledge is crucial.

Application of CRISPR Interference (CRISPRi) for Gene Silencing in Pathogenic Species of Leptospira.

Leptospirosis is a global neglected zoonosis, responsible for at least 1 million cases per year and almost 60 thousand deaths. The disease is caused by pathogenic and virulent bacteria of the genus Leptospira, either by direct contact with the bacteria or indirectly by exposure to contaminated water or soil. Domestic and wild animals act as reservoir hosts of infection, shedding leptospires from colonized renal tubules of the kidney, via urine, into the environment. The generation of mutant strains of Leptospira is critical to evaluate and understand pathogenic mechanisms of infection. CRISPR interference (CRISPRi) has proven to be a straightforward, affordable, and specific tool for gene silencing in pathogenic Leptospira. Therefore, the methodological details of obtaining the plasmid constructs containing both dCas9 and guide RNA, delivery of plasmids to Leptospira by conjugation with the E. coli strain ß2163, and transconjugant recovery and evaluation, will be described. In addition, the recently described Hornsby-Alt-Nally (HAN) media allows for the relatively rapid isolation and selection of mutant colonies on agar plates.

Cancer of unknown primary in a mare: case report and comparative pathology review.

A 25-y-old Percheron mare was admitted to the teaching hospital because of lethargy and intractable dyspnea. Thoracoabdominal ultrasound examination identified severe peritoneal effusion, mild bilateral pleural effusion, and a diffuse pulmonary nodular pattern. Cytology of peritoneal fluid revealed a hypercellular sample with clusters of neoplastic polygonal cells and admixed macrophages. Euthanasia was followed by postmortem examination; marked bi-cavitary effusion was present, and innumerable up to 4-cm diameter, round-to-floriform nodules were diffusely evident throughout serosal surfaces as well as the pulmonary and hepatic parenchyma. Disseminated adenocarcinoma, predominantly affecting lung and liver with widespread serosal implantation, was confirmed on light microscopy. Neoplastic cells had strong immunolabeling for pancytokeratin and lacked immunoreactivity to vimentin, napsin A, and Pax8. Cytokeratin 7 and thyroid transcription factor-1 were non-contributory given absent and inconsistent internal control reactivity, respectively. Such results, combined with the lack of a major mass that would indicate a primary site, were supportive of carcinoma of unknown primary site, which remains a conundrum in human oncology, and is poorly explored in veterinary medicine, mainly as a result of clinical and diagnostic limitations.

Genetic manipulation of pathogenic Leptospira: CRISPR interference (CRISPRi)-mediated gene silencing and rapid mutant recovery at 37 °C.

Leptospirosis is a neglected, widespread zoonosis caused by pathogenic species of the genus Leptospira, and is responsible for 60,000 deaths per year. Pathogenic mechanisms of leptospirosis remain poorly understood mainly because targeted mutations or gene silencing in pathogenic Leptospira continues to be inherently inefficient, laborious, costly and difficult to implement. In addition, pathogenic leptospires are highly fastidious and the selection of mutants on solid agar media can take up to 6 weeks. The catalytically inactive Cas9 (dCas9) is an RNA-guided DNA-binding protein from the Streptococcus pyogenes CRISPR/Cas system and can be used for gene silencing, in a strategy termed CRISPR interference (CRISPRi). Here, this technique was employed to silence genes encoding major outer membrane proteins of pathogenic L. interrogans. Conjugation protocols were optimized using the newly described HAN media modified for rapid mutant recovery at 37 °C in 3% CO2 within 8 days. Complete silencing of LipL32 and concomitant and complete silencing of both LigA and LigB outer membrane proteins were achieved, revealing for the first time that Lig proteins are involved in pathogenic Leptospira serum resistance. Gene silencing in pathogenic leptospires and rapid mutant recovery will facilitate novel studies to further evaluate and understand pathogenic mechanisms of leptospirosis.

Phoretic mites (Rhinoseius spp. ) in Apodiformes from Cerrado and Pantanal Biomes in midwestern Brazil / Ácaros Foréticos (Rhinoseius spp. ) em Apodiformes dos Biomas Cerrado e Pantanal no Centro-Oeste do Brasil

Abstract Nasal mites are commonly found in hummingbirds (Apodiformes). In most cases, endoparasitic mites were reported to parasitize the respiratory system, particularly the anterior nasal chambers, larynx, trachea, lungs, and the air and conjunctival sacs. However, some mites are only carried by hummingbirds, because flowers serving as a nutrient source for these small mites, such as for Apodiformes, a tropical nectar-feeding in their natural habitat. From August 2012 to May 2014, humminbirds were captured using mist nets, and examined for the presence of nasal mites (identified according to specific keys) in the Pantanal, and Cerrado biomes, from the Mato Grosso state, midwestern Brazil. Overall, 76 Apodiform specimens, from the Trochilidae family were captured, and 20 female nasal mites, identified as being from the Rhinoseius genus, were collected from eight hosts (10.5%), of the following species: Amazilia fimbriata, Amazilia versicolor, Eupetonema macroura, and Thalurania furcata. The Rhinoseius spp. has been reported on, in many ecosystems in the Americas, including Brazil, however, this is the first report on Apodiformes from the Pantanal biome. Phoretic mites are not responsible for direct damage to the population of Apodiformes, as they are not pathogenic. However, they compete for food with hummingbirds and decrease the quantity of food available in the environment. They also affect the reproductive dynamics of plants, feed on pollen, and interfere with the intricacies of the biodiversity where they live.

Resumo Os ácaros nasais são comumente encontrados em beija-flores (Apodiformes). Na maioria dos casos, os ácaros foram relatados como endoparasitas, parasitando o sistema respiratório, particularmente as câmaras nasais anteriores, a laringe, a traqueia, os pulmões e os sacos aéreos e conjuntivais. No entanto, alguns ácaros são transportados apenas por beija-flores, pois flores servem como fonte de nutrientes para esses pequenos ácaros, assim como para Apodiformes tropicais alimentados de néctar em seu habitat natural. De agosto de 2012 a maio de 2014, beija-flores foram capturadas com redes de neblina e examinadas quanto à presença de ácaros nasais (identificados de acordo com chaves específicas) nos biomas Pantanal e Cerrado, no estado do Mato Grosso, Centro-Oeste do Brasil. No total, 76 exemplares Apodiformes da família Trochilidae foram capturados e 20 fêmeas de ácaros nasais, identificados como pertencentes ao gênero Rhinoseius, foram coletados de oito hospedeiros (10,5%), das seguintes espécies: Amazilia fimbriata, Amazilia versicolor, Eupetonema macroura e Thalurania furcata. Rhinoseius spp. foi relatado em muitos ecossistemas nas Américas, incluindo o Brasil, no entanto, este é o primeiro relato em Apodiformes do bioma Pantanal. Os ácaros foréticos não são responsáveis ​​por danos diretos à população de Apodiformes, pois não são patogênicos. No entanto, eles competem por comida com beija-flores e diminuem a quantidade de comida disponível no ambiente. Eles também afetam a dinâmica reprodutiva das plantas, se alimentam de pólen e interferem nas interações da biodiversidade em que vivem.