Lineage Plasticity in SCLC Generates Non-Neuroendocrine Cells Primed for Vasculogenic Mimicry.

INTRODUCTION: Vasculogenic mimicry (VM), the process of tumor cell transdifferentiation to endow endothelial-like characteristics supporting de novo vessel formation, is associated with poor prognosis in several tumor types, including SCLC. In genetically engineered mouse models (GEMMs) of SCLC, NOTCH, and MYC co-operate to drive a neuroendocrine (NE) to non-NE phenotypic switch, and co-operation between NE and non-NE cells is required for metastasis. Here, we define the phenotype of VM-competent cells and molecular mechanisms underpinning SCLC VM using circulating tumor cell-derived explant (CDX) models and GEMMs. METHODS: We analyzed perfusion within VM vessels and their association with NE and non-NE phenotypes using multiplex immunohistochemistry in CDX, GEMMs, and patient biopsies. We evaluated their three-dimensional structure and defined collagen-integrin interactions. RESULTS: We found that VM vessels are present in 23/25 CDX models, 2 GEMMs, and in 20 patient biopsies of SCLC. Perfused VM vessels support tumor growth and only NOTCH-active non-NE cells are VM-competent in vivo and ex vivo, expressing pseudohypoxia, blood vessel development, and extracellular matrix organization signatures. On Matrigel, VM-primed non-NE cells remodel extracellular matrix into hollow tubules in an integrin ß1-dependent process. CONCLUSIONS: We identified VM as an exemplar of functional heterogeneity and plasticity in SCLC and these findings take considerable steps toward understanding the molecular events that enable VM. These results support therapeutic co-targeting of both NE and non-NE cells to curtail SCLC progression and to improve the outcomes of patients with SCLC in the future.

Proteomics analysis of urine and catheter-associated biofilms in spinal cord injury patients.

After spinal cord injury (SCI), use chronic urinary catheters for bladder management is common, making these patients especially vulnerable to catheter-associated complications. Chronic catheterization is associated with bacterial colonization and frequent catheter-associated urinary tract infections (CAUTI). One determinant of infection success and treatment resistance is production of catheter-associated biofilms, composed of microorganisms and host- and microbial-derived components. To better understand the biofilm microenvironment, we performed proteomics analysis of catheter-associated biofilms and paired urine samples from four people with SCI with chronic indwelling urinary catheters. We developed a novel method for the removal of adhered cellular components on catheters that contained both human and microbial homologous proteins. Proteins from seven microbial species were identified including: Escherichia coli, Klebsiella species (spp), Enterococcus spp, Proteus mirabilis, Pseudomonas spp, Staphylococcus spp, and Candida spp. Peptides identified from catheter biofilms were assigned to 4,820 unique proteins, with 61% of proteins assigned to the biofilm-associated microorganisms, while the remainder were human-derived. Contrastingly, in urine, only 51% were assigned to biofilm-associated microorganisms and 4,554 proteins were identified as a human-derived. Of the proteins assigned to microorganisms in the biofilm and paired urine, Enterococcus, Candida spp, and P. mirabilis had greater associations with the biofilm phase, whereas E. coli and Klebsiella had greater associations with the urine phase, thus demonstrating a significant difference between the urine and adhered microbial communities. The microbial proteins that differed significantly between the biofilm and paired urine samples mapped to pathways associated with amino acid synthesis, likely related to adaptation to high urea concentrations in the urine, and growth and protein synthesis in bacteria in the biofilm. Human proteins demonstrated enrichment for immune response in the catheter-associated biofilm. Proteomic analysis of catheter-associated biofilms and paired urine samples has the potential to provide detailed information on host and bacterial responses to chronic indwelling urinary catheters and could be useful for understanding complications of chronic indwelling catheters including CAUTIs, urinary stones, and catheter blockages.

Oncogene-mediated metabolic gene signature predicts breast cancer outcome.

Breast cancer remains the second most lethal cancer among women in the United States and triple-negative breast cancer is the most aggressive subtype with limited treatment options. Trop2, a cell membrane glycoprotein, is overexpressed in almost all epithelial cancers. In this study, we demonstrate that Trop2 is overexpressed in triple-negative breast cancer (TNBC), and downregulation of Trop2 delays TNBC cell and tumor growth supporting the oncogenic role of Trop2 in breast cancer. Through proteomic profiling, we discovered a metabolic signature comprised of TALDO1, GPI, LDHA, SHMT2, and ADK proteins that were downregulated in Trop2-depleted breast cancer tumors. The identified oncogene-mediated metabolic gene signature is significantly upregulated in TNBC patients across multiple RNA-expression clinical datasets. Our study further reveals that the metabolic gene signature reliably predicts poor survival of breast cancer patients with early stages of the disease. Taken together, our study identified a new five-gene metabolic signature as an accurate predictor of breast cancer outcome.

Reconstructed Apoptotic Bodies as Targeted "Nano Decoys" to Treat Intracellular Bacterial Infections within Macrophages and Cancer Cells.

Staphylococcus aureus (S. aureus) is a highly pathogenic facultative anaerobe that in some instances resides as an intracellular bacterium within macrophages and cancer cells. This pathogen can establish secondary infection foci, resulting in recurrent systemic infections that are difficult to treat using systemic antibiotics. Here, we use reconstructed apoptotic bodies (ReApoBds) derived from cancer cells as "nano decoys" to deliver vancomycin intracellularly to kill S. aureus by targeting inherent "eat me" signaling of ApoBds. We prepared ReApoBds from different cancer cells (SKBR3, MDA-MB-231, HepG2, U87-MG, and LN229) and used them for vancomycin delivery. Physicochemical characterization showed ReApoBds size ranges from 80 to 150 nm and vancomycin encapsulation efficiency of 60 ± 2.56%. We demonstrate that the loaded vancomycin was able to kill intracellular S. aureus efficiently in an in vitro model of S. aureus infected RAW-264.7 macrophage cells, and U87-MG (p53-wt) and LN229 (p53-mt) cancer cells, compared to free-vancomycin treatment (P < 0.001). The vancomycin loaded ReApoBds treatment in S. aureus infected macrophages showed a two-log-order higher CFU reduction than the free-vancomycin treatment group. In vivo studies revealed that ReApoBds can specifically target macrophages and cancer cells. Vancomycin loaded ReApoBds have the potential to kill intracellular S. aureus infection in vivo in macrophages and cancer cells.

Evaluation of a Chinese herbal supplement on equine squamous gastric disease and gastric fluid pH in mares.

BACKGROUND: Wei Le San (WLS) is a Chinese herbal formula comprised of 9 herbs selected for their putative anti-inflammatory effects. OBJECTIVES: To evaluate the effects of WLS administration in horses with nonglandular gastric ulcers. ANIMALS: Ten mixed breed mares (aged 7-21 years, 401-567 kg body weight). METHODS: Experimental design was a blinded, prospective, 2-period crossover study. All horses received a placebo (25 mL dextrose-based syrup; n = 10) and the treatment (WLS, 5 g in 25 mL dextrose-based syrup; n = 10), administered twice daily. Horses underwent a 1-week, alternating feed-deprivation period to induce or worsen existing ulcers; treatment began on day 7. Gastroscopic examination was performed on d0, d6, and d35, with gastric fluid pH obtained on d6 and d35. Gastric ulcer scores assigned by 3 masked observers were averaged for each examination. RESULTS: Ulcer number scores for horses treated with WLS (median = 0; range, 0-4) was not different from the untreated controls (median = 0.5; range, 0-4; P = .81) by the end of the treatment period. Ulcer severity score for treated horses (median = 0; range, 0-1) was also unchanged compared to the control group (median = 0.5; range, 0-1; P = .85). Gastric pH was not altered by either treatment, with a median of 2.1 (range, 1.9-4.1) for the horses treated with WLS and 2.8 (range, 1.6-7.2) in the untreated controls (P = .46). CONCLUSIONS AND CLINICAL IMPORTANCE: The experimental model used to induce gastric ulceration was unable to discern a difference between the herbal supplement and the placebo in normal horses.

The effect of foot-and-mouth disease vaccination on early pregnancy loss in beef heifers in Argentina.

The threat of Foot and Mouth Disease (FMD) in South America has global economic implications and retaining a FMD Free status under the World Organization for Animal Health (OIE) remains a top priority. In Argentina the Servicio Nacional de Sanidad y Calidad Agroalimentaria (SENASA), the national service of agri-food health and quality, requires cattle located in the Pampean region of the Salado River basin to receive two foot-and-mouth disease (FMD) vaccinations per year, which results in one vaccination coinciding with beef cattle breeding season. While the vaccination program remains necessary, there is a growing concern amongst food animal veterinarians, that the overlap of FMD vaccination with the first 35 days of the breeding season is associated with early pregnancy loss (EPL). To address this concern, a preliminary randomized controlled trial t study was conducted to investigate the risk ratio (RR) of EPL in vaccinated, pregnant Aberdeen Angus heifers. Initially (Day 0), 858 heifers underwent fixed time-AI (FTAI). Subsequently, on day 33, following pregnancy diagnosis by transrectal ultrasonography pregnant heifers (n = 311) were randomly allocated to two treatment groups. Group 1 (162 animals) received an inactivated oil emulsion FMD vaccine, and Group 2 (149 animals) received a saline injection (control). On day 51 (18 days post vaccination), pregnancy status was re-evaluated by ultrasonography. The initial pregnancy rate (PR) on Day 33 was 58% (498/858 animals). On Day 51 (18 days post vaccination), PR in Group 1 was 96.3% (156/162 animals), and in Group 2 (control) was 98.6% (147/149 animals). The EPL in Group 1 was 3.7% (6/162 animals) and in Group 2 was 1.3% (2/149 animals). The RR of EPL in Group 1, compared to Group 2, was 2.8 (95% confidence interval: 0.6-13, p-value: 0.20). With such a wide range in confidence intervals and a p value of 0.20 a larger prospective study would be necessary to establish an unequivocally statistically significant link between heifer vaccination 33 days post FTAI and an increased risk of EPL.

Equine motor neuron disease in 2 horses from Saskatchewan.

Two horses from Saskatchewan were presented with signs of sweating, muscle fasciculations, weight loss, and generalized weakness. The horses were diagnosed with equine motor neuron disease (EMND), by histological assessment of a spinal accessory nerve or sacrocaudalis dorsalis medialis muscle biopsy. This is the first report of EMND in western Canada.

Maladie équine des motoneurones chez 2 chevaux de la Saskatchewan. Deux chevaux de la Saskatchewan ont été présentés avec des signes de sudation, de fibrillations musculaires, de perte de poids et de faiblesse généralisée. On a diagnostiqué la maladie équine des motoneurones (MEMN) chez les chevaux en effectuant une évaluation histologique d'un nerf accessoire de la colonne vertébrale ou une biopsie du muscle sacrocaudalis dorsalis medialis. Il s'agit du premier rapport de MEMN dans l'Ouest canadien.(Traduit par Isabelle Vallières).

Detection of Neorickettsia risticii, the agent of Potomac horse fever, in a Gypsy Vanner stallion from Manitoba.

A horse with colitis from Manitoba referred to the Veterinary Medical Centre, Western College of Veterinary Medicine, was diagnosed with Potomac horse fever (PHF). Polymerase chain reaction analysis of the feces confirmed Neorickettsia risticii infection. This is the first reported case of PHF in Manitoba.

Détection deNeorickettsia risticii,agent de la fièvre du Potomac, chez un étalon Gypsy Vanner du Manitoba. Un diagnostic de fièvre du Potomac a été posé pour un cheval du Manitoba atteint de colite qui avait été recommandé au Veterinary Medical Centre du Western College of Veterinary Medicine. Une analyse des fèces par amplification en chaîne par la polymérase a confirmé l'infection par Neorickettsia risticii. Il s'agit du premier cas de fièvre du Potomac au Manitoba.(Traduit par Isabelle Vallières).

Effect of rider experience and evaluator expertise on subjective grading of lameness in sound and unsound sports horses under saddle.

The primary objective of this study was to investigate whether rider experience influences the assessment and grading of lameness in horses based on under-saddle gait analysis. Thirteen adult sports horses in active training were included in the study. After a baseline lameness and neurologic examination by the principal investigators, horses were videotaped while being ridden by an experienced and a less experienced rider. A 3-minute video was made for each horse and rider and 26 videos were randomly ordered and compiled on a DVD. Veterinarians with different levels of experience in evaluating lameness and veterinary students viewed the DVD and assigned a lameness score to each horse/rider combination. In a model accounting for the expertise of the evaluator, there was no difference in overall lameness scores between experienced and less experienced riders. This result was consistent for both sound and unsound horses. The overall lameness scores reported by specialists and students, however, differed significantly. The lameness score reported by the study participants while the horse was ridden was significantly associated with the subjective baseline lameness assessment reported by the principal investigators for the same limb when the horse was not under saddle. Additional work is necessary to determine whether riders with even lower skill levels would further alter the balance and motion pattern of the horse and have more influence on subjective grading of lameness.

L'objectif principal de la présente étude était d'examiner si l'expérience du cavalier influence l'évaluation et la gradation de la boiterie chez des chevaux basées sur l'analyse de la démarche sous-selle. Treize chevaux sportifs adultes en entraînement actif ont été inclus dans l'étude. Suite à un examen neurologique et de boiterie de base par les chercheurs principaux, les chevaux ont été filmés alors qu'ils étaient montés par un cavalier d'expérience et un cavalier moins expérimenté. Une vidéo de 3 minutes a été réalisée pour chaque combinaison cheval/cavalier et les 26 vidéos ont été compilées de manière aléatoire sur un DVD. Des vétérinaires ayant différents niveaux d'expérience à évaluer les boiteries et des étudiants vétérinaires ont visualisé le DVD et donné un pointage à chaque combinaison cheval/cavalier. Dans un modèle tenant compte de l'expérience de l'évaluateur, il n'y avait aucune différence dans les pointages globaux entre un cavalier expérimenté et un moins expérimenté. Ce résultat était constant autant pour les chevaux solides que fragiles. Toutefois, les pointages globaux de boiterie notés par les spécialistes et les étudiants étaient significativement différents. Le pointage de boiterie rapporté par les participants à l'étude alors que le cheval était conduit était significativement associé avec l'évaluation subjective de base rapportée par les chercheurs principaux pour le même membre lorsque le cheval n'était pas sous-selle. Des études additionnelles sont nécessaires afin de déterminer si des cavaliers avec encore moins d'expérience influenceraient encore plus l'équilibre et le patron de mouvement du cheval et aurait plus d'influence sur la gradation subjective de la boiterie.(Traduit par Docteur Serge Messier).

Failure of a VapA/CpG oligodeoxynucleotide vaccine to protect foals against experimental Rhocococcus equi pneumonia despite induction of VapA-specific antibody and interferon-γ response.

We evaluated the immunogenic and protective potential of a recombinant VapA/CpG oligodeoxynucleotide (ODN) 2395 vaccine in neonatal foals undergoing experimental Rhodococcus equi challenge. Foals (n = 8) were vaccinated by intramuscular injection on days 1 and 15 of the study; control foals (n = 7) received a phosphate-buffered saline (PBS) solution. All foals were challenged by intrabronchial administration of 5 × 106 R. equi 103⁺ on day 29. Bronchoalveolar lavages were done on days 15, 29, and 36 and total cell count, differential cell count, rVapA-stimulated cell proliferation and interferon (IFN)-γ mRNA expression determined. Clinical examination, complete blood (cell) counts, serology for VapA-specific antibodies, and culture of nasal and fecal swabs were done on days 1, 15, 29, 36, 43, and 50. Foals were humanely euthanized on day 50 and severity of pneumonia scored on a 4-point scale. Vaccination resulted in a significant increase in VapA-specific immunoglobulin (Ig) production, with total IgG and IgG(T) being increased by day 15. Expression of VapA-specific IFN-γ mRNA by BAL cells was increased in the vaccinated foals following challenge. Postmortem lung severity scores did not differ between groups. Two foals shed virulent R. equi in feces; however, real-time polymerase chain reaction (PCR) revealed the isolates to be different from the challenge strain.

Nous avons évalué le potentiel immunogène et protecteur d'un vaccin recombinant VapA/oligodéoxynucléotide CpG (ODN) 2395 chez des poulains nouveau-nés soumis à une infection défi par Rhodococcus equi. Les poulains (n = 8) étaient vaccinés par voie intramusculaire aux jours 1 et 15 de l'étude; les poulains témoins (n = 7) ont reçu une injection d'une solution de saline tamponnée (PBS). Tous les poulains ont été challengés par administration intra-bronchique de 5 × 106R. equi 103+ au jour 29. Des lavages broncho-alvéolaires (LBA) ont été effectués aux jours 15, 29 et 36 et on détermina le nombre total de cellules, un dénombrement cellulaire différentiel, la prolifération des cellules rVapA stimulées et l'expression d'ARNm de l'interféron (IFN)-γ. Un examen clinique, des comptages cellulaires sanguins complets, une analyse sérologique pour détecter les anticorps spécifiques contre VapA, et une culture d'écouvillons nasal et fécal ont été effectués aux jours 1, 15, 29, 36, 43 et 50. Les poulains ont été euthanasiés au jour 50 et la sévérité de la pneumonie notée sur une échelle de 4 points. La vaccination a causé une augmentation significative de la production d'immunoglobulines (Ig) spécifiquement dirigées contre VapA, les quantités totales d'IgG et d'IgG(T) ayant augmentées au jour 15. L'expression d'ARNm de l'IFN-γ spécifique au VapA par les cellules des LBA était augmentée chez les poulains vaccinés suite au challenge. Aucune différence ne fut notée dans les pointages de sévérité des lésions pulmonaires lors des examens post-mortem. Deux poulains excrétaient du R. equi virulent dans leurs fèces; toutefois, l'analyse par réaction d'amplification en chaîne par la polymérase (PCR) a démontré que ces isolats étaient différents de la souche utilisée pour le challenge.(Traduit par Docteur Serge Messier).

Investigation of a progressive facial deformity and stridor in an adult horse with a bone sequestrum and subsequent sinonasal abscess.

Advanced diagnostic imaging and histopathological investigation were performed in an adult horse with chronic facial swelling due to a bone sequestrum and abscessation. In contrast to other cases, there were no draining tracts, difficulty eating, weight loss, head shaking, or fistula formation between the oral cavity and the maxillary sinus.

Respiratory distress due to retropharyngeal and neck swelling in a horse with mediastinal lymphosarcoma.

A 9-year-old, 1494-lb (679-kg) Quarter horse gelding in good body condition was admitted to the Veterinary Teaching Hospital, Western College of Veterinary Medicine (WCVM), University of Saskatchewan, for evaluation and treatment of acute respiratory distress. On a trail ride 26 days before presentation, the gelding had been exposed to a horse with clinical signs of an upper respiratory tract infection. The gelding performed well on the trail ride but, 4 days later, developed a mild swelling in the retropharyngeal area and a slight bilateral mucoid nasal discharge. The gelding was examined at the farm by its primary veterinarian, who made a presumptive diagnosis of strangles. Treatment was initiated using a combination of trimethoprim/sulfamethoxazole (30 mg/kg PO q12h for 1 week) and phenylbutazone (6 mg/kg PO q24h for the first 2 days, followed by 3.5 mg/kg PO q24h for 5 more days). The gelding did not respond satisfactorily to antimicrobial and antiinflammatory therapy. During the following 2 weeks, the retropharyngeal swelling gradually became more pronounced and extended to the upper portion of the neck. Two days before presentation at WCVM, the gelding also developed a dry cough with difficult breathing, and the retropharyngeal and neck swelling extended to the pectoral region and ventral pectoral area extending to the cranial ventral abdomen. Because of progressing clinical signs and poor response to treatment, the horse was referred to WCVM for further evaluation.

Pemphigus foliaceus in a juvenile Cashmere goat, and outcome after prednisolone and methylprednisolone therapy.

A juvenile Cashmere goat was evaluated and treated for generalized pustular and crusting dermatitis. A diagnosis of pemphigus foliaceus was made based on histopathology and immunohistochemistry. Treatment with prednisolone resulted in resolution of the lesions. Despite tapering doses and combination with methylprednisolone for monthly maintenance, the kid was euthanized for poor growth and fiber quality.