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Translationally relevant animal models are essential for the successful translation of basic science findings into clinical medicine. While rodent models are widely accessible, there are numerous limitations that prevent the extrapolation of findings to human medicine. One approach to overcome these limitations is to use animal models that are genetically diverse and naturally develop disease. For example, pet dogs spontaneously develop diseases that recapitulate the natural progression seen in humans and live in similar environments alongside humans. Thus, dogs represent a useful animal model for many areas of research. Despite the value of the canine model, species specific reagent limitations have hampered in depth characterization of canine immune cells, which constrains the conclusions that can be drawn from canine immunotherapy studies. To address this need, we used single-cell RNA sequencing to characterize the heterogeneity of circulating leukocytes in healthy dogs (n = 7) and osteosarcoma (OS) affected dogs (n = 10). We present a cellular atlas of leukocytes in healthy dogs, then employ the dataset to investigate the impact of primary OS tumors on the transcriptome of circulating leukocytes. We identified 36 unique cell populations amongst dog circulating leukocytes, with a remarkable amount of heterogeneity in CD4 T cell subtypes. In our comparison of healthy dogs and dogs with OS, we identified relative increases in the abundances of polymorphonuclear (PMN-) and monocytic (M-) myeloid-derived suppressor cells (MDSCs), as well as aberrations in gene expression within myeloid cells. Overall, this study provides a detailed atlas of canine leukocytes and investigates how the presence of osteosarcoma alters the transcriptional profiles of circulating immune cells.

Canine acute leukaemia is a heterogeneous neoplasm with multiple phenotypes. Criteria to subtype acute leukaemia by flow cytometry have not been validated. The goal of this study was to develop a panel of antibodies and objective antigen expression criteria for the assignment of lymphoid or myeloid lineage by flow cytometry. We isolated mRNA from the blood of 45 CD34+ acute leukaemia cases and measured expression of 43 genes that represent lymphoid and myeloid lineages using NanoString technology. We determined differentially expressed genes between major groups identified by unsupervised hierarchical clustering. We then evaluated the expression of antigens by flow cytometry to determine if cases could be assigned to a lineage. Two groups were identified by gene expression. Group 1/LYMPH overexpressed lymphoid-associated genes (ex. DNTT) and had a higher percentage of CD5 + CD3- cells by flow cytometry. Group 2/MYELO overexpressed myeloid-associated genes (ex. ANPEP/CD13) and had a higher percentage of class II major histocompatibility complex (MHCII)- CD14+ and/or CD18 + CD4- cells. We proposed that12.5% CD5 + CD3- cells in the blood was indicative of lymphoid lineage, and 3.0% CD14 + MHCII- cells or 18% CD18 + MHCII-CD4- cells was indicative of myeloid lineage. 15/15 cases that met the proposed criteria for acute lymphocytic leukaemia were in LYMPH group and 12/15 cases that met the proposed criteria for acute myeloid leukaemia were in MYELO group. The majority of CD34+ cases that did not meet either immunophenotyping lineage criterion (12/13) clustered within the LYMPH group. In conclusion, currently available antibodies can be useful for determining canine acute leukaemia subtypes.

BACKGROUND Densitometric quantification of myeloma paraproteins (M-proteins) is used to monitor secretory myeloma related disorders in humans and dogs. The previous work in dogs used agarose gel electrophoresis (AGE) but did not establish if other methods of serum protein electrophoresis, such as capillary zone electrophoresis (CZE), were comparable. OBJECTIVES We aimed to determine if the densitometric quantification of M-proteins using CZE would yield results comparable to AGE methods. METHODS Fifty-one serum samples from 22 dogs and 18 cats with confirmed monoclonal gammopathies and previously performed AGE were evaluated using CZE on a Sebia Minicap system. Samples were run in duplicate, and their M-proteins were densitometrically measured using the corrected perpendicular drop method previously described. Human-based quality control samples were used to determine the inter-run coefficient of variation (CV). Patient samples were used to calculate the intra-run CV. Method comparison was performed using simple linear regression, Passing-Bablok regression, and Bland-Altman analyses, and Medx evaluations. RESULTS Inter-run and intra-run CVs for CZE were 3.71%-7.65% and 2.89%-4.74%, respectively. Simple linear regression demonstrated an excellent correlation (r > 0.98). Passing-Bablok regression was compatible with the presence of proportional bias in the entire population, and Bland-Altman plots revealed a proportional bias in the feline cases. The Medx evaluation suggested that the two methods did not perform similarly in clinical samples with poor performance at a decision limit of 0.5 gm/dL. CONCLUSIONS Capillary zone electrophoresis is an acceptable method for M-protein densitometric quantification in canine and feline sera but cannot be used interchangeably with AGE-based evaluations.

Colorectal cancer (CRC) is the second most prevelant malignancy in Europe and diet is an important modifiable risk factor. Processed meat consumption, including meats with preservative salts such as sodium nitrite, have been implicated in CRC pathogenesis. This study investigated how the CRC pathology and metabolic status of adenomatous polyposis coli (APC) multiple intestinal neoplasia (min) mice was perturbed following 8 weeks of pork meat consumption. Dietary inclusions (15%) of either nitrite-free pork, nitrite-free sausage, or nitrite-containing sausage (frankfurter) were compared against a parallel control group (100% chow). Comprehensive studies investigated: gastrointestinal tract histology (tumours), aberrant crypt foci (ACF), mucin deplin foci (MDF), lipid peroxidation (urine and serum), faecal microbiota, and serum metabolomics (599 metabolites). After 8 weeks mice consuming the frankfurter diet had 53% more (P = 0.014) gastrointestinal tumours than control, although ACF and MDF did not differ. Urine and serum lipid peroxidation markers were 59% (P = 0.001) and 108% (P = 0.001) higher, respectively in the frankfurter group. Gut dysbiosis was evident in these mice with comparably fewer Bacteriodes and more Firmicutes. Fasting serum levels of trimethylamine N-oxide (TMAO) and numerous triglycerides were elevated. Various serum phosphotidylcholine species were decreased. These results demonstrate that nitrite-containing sausages may exaccerbate the development of CRC pathology in APCMin mice to a greater extent than nitrite-free sausages, and this is associated with greater lipid peroxidation, wide-ranging metabolic alternation and gut dysbiosis.

Ipatasertib is a selective, small molecule Akt inhibitor that is currently being developed for the treatment of metastatic castration-resistant prostate cancer. Darolutamide is an androgen receptor (AR) inhibitor that is approved for the treatment of non-metastatic castration-resistant prostate cancer. Ipatasertib is metabolized by CYP3A4 to form a less active metabolite M1 (G-037720). Ipatasertib is also a weak time-dependent CYP3A4 inhibitor. Darolutamide is a mild CYP3A4 inducer and is metabolized into an active keto-darolutamide metabolite via CYP3A4. In this Phase 1b open-label, single sequence crossover study, ipatasertib pharmacokinetics safety and tolerability were evaluated in combination with darolutamide in metastatic castration-resistant prostate cancer (n = 15 patients). Specifically, the effect of 600 mg BID of darolutamide on 400 mg QD ipatasertib was evaluated in this study. Based on pharmacokinetic analysis, a mild reduction in ipatasertib AUC0–24 h,ss and Cmax,ss exposures was observed (~8% and ~21%, respectively) when administered in combination with darolutamide, which is considered not clinically meaningful. M1 exposures were similar with and without darolutamide administration. Darolutamide and keto-darolutamide exposures in combination with ipatasertib were similar to previously reported exposures for single agent darolutamide. Overall, the combination appears to be well-tolerated in the metastatic castration-resistant prostate cancer indication with very few AEs.

Loss PTEN function is one of the most common events driving aggressive prostate cancers and biochemically, PTEN is a lipid phosphatase which opposes the activation of the oncogenic PI3K-AKT signalling network. However, PTEN also has additional potential mechanisms of action, including protein phosphatase activity. Using a mutant enzyme, PTEN Y138L, which selectively lacks protein phosphatase activity, we characterised genetically modified mice lacking either the full function of PTEN in the prostate gland or only lacking protein phosphatase activity. The phenotypes of mice carrying a single allele of either wild-type Pten or PtenY138L in the prostate were similar, with common prostatic intraepithelial neoplasia (PIN) and similar gene expression profiles. However, the latter group, lacking PTEN protein phosphatase activity additionally showed lymphocyte infiltration around PIN and an increased immune cell gene expression signature. Prostate adenocarcinoma, elevated proliferation and AKT activation were only frequently observed when PTEN was fully deleted. We also identify a common gene expression signature of PTEN loss conserved in other studies (including Nkx3.1, Tnf and Cd44). We provide further insight into tumour development in the prostate driven by loss of PTEN function and show that PTEN protein phosphatase activity is not required for tumour suppression.

Background Posttraumatic stress disorder (PTSD) is a pervasive mental health condition with limited treatment success. Transcranial magnetic stimulation (TMS) has shown positive outcomes for people with PTSD, using different treatment protocols. This meta-analysis sought to examine which variables in TMS treatment are associated with treatment benefits. Methods A literature search of major online research databases from inception to September 15, 2020 was conducted to identify primary research studies using TMS to treat PTSD. Treatment effect data and TMS treatment variables were coded and analysed using a random effects model. Meta-regression and analyses of moderating variables were conducted to ascertain which variables were associated with significant treatment effects. Results An overall effect size of d = 1.17, 95% CI [0.89 – 1.45] for TMS as a treatment for PTSD was found. Analysis of moderators showed that there was a significantly larger treatment effect for high frequency TMS (d = 1.44) compared with low frequency (d = 0.72), p = .006; there was no significant difference between TMS targeting the left dorsolateral prefrontal cortex (DLPFC) and the right DLPFC; and larger treatment doses were not associated with stronger treatment effects. Limitations Not all published studies were available in English or reported the necessary data to be included in this meta-analysis. Conclusions TMS shows potential as a treatment for PTSD, although further research is required to understand the neurological mechanisms of TMS on specific PTSD symptoms so that more effective treatment can be designed for individuals.

Previous research has demonstrated that reversing the contrast of the eye region, which includes the eyebrows, affects the N170 ERP. To selectively assess the impact of just the eyes, the present study evaluated the N170 in response to reversing contrast polarity of just the iris and sclera in upright and inverted face stimuli. Contrast reversal of the eyes increased the amplitude of the N170 for upright faces, but not for inverted faces, suggesting that the contrast of eyes is an important contributor to the N170 ERP.

Routine electrophoresis [agarose gel electrophoresis (AGE) and capillary zone electrophoresis (CZE)] and species-specific immunofixation (IF) can be used alone or in combination to detect immunoglobulin paraprotein (M-protein) and diagnose secretory myeloma-related disorders (sMRD).We aimed to evaluate the performance of AGE, CZE, CZE plus IF (CZE-IF), and AGE plus IF (AGE-IF) for detecting canine serum M-proteins.One hundred canine cases that had AGE, CZE, and routine IF performed on serum, and where B-cell lineage neoplasia (such as B-cell lymphoma and plasma cell tumors) had been diagnosed or excluded, were evaluated. Routine IF protocols targeted IgG-FC, IgA, and IgM heavy chains and light chains. IgG4 IF and free light chain IF were also performed. B-cell lineage neoplasms with an M-protein detected, using any available method, were classified as sMRD. Datasets from AGE, CZE, IF, CZE-IF, and AGE-IF (electrophoretograms, gel images, and fraction concentrations) were composed and reviewed. The sensitivity, specificity, and Youden's index for M-protein detection were determined for each dataset.The combination of AGE-IF or CZE-IF was more sensitive (82.9%) than CZE alone (72.0%) or AGE alone (64.6%) and more specific (66.1%, 48.3%, 51.7%, respectively). Immunofixation could be used alone to detect M-proteins (sensitivity 82.9%, specificity 61.9%), but there were technical challenges that complicated the performance and evaluation of the test. Myeloma with free light chains only was found in 5/41 cases of sMRD.Adding routine IF to routine electrophoresis increases the ability to accurately identify M-proteins; however, there is still room for further diagnostic performance improvements.

Background Detection of free light chains (fLC) in animals relies on protein electrophoresis or the Bence-Jones protein test on urine. Objective To describe the detection of both serum fLC (sfLC) and urine fLC (ufLC) in 8 dogs and 2 cats using a commercially available human immunofixation (IF) kit. Animals Archived serum or urine samples from 27 dogs and 2 cats submitted to the Colorado State University Veterinary Diagnostic Laboratory for routine diagnostics. Methods Retrospective study evaluating the presence of fLC in dogs and cats using agarose gel electrophoresis and routine and fLC IF performed on serum and urine. The performance of the fLC IF reagents was evaluated using samples characterized by routine IF, tandem mass spectrometry, and a combination of fLC IF and western blotting. Free light chains were documented by paired electrophoresis and fLC IF. Results The fLC only myeloma case developed end-stage renal failure 5 months post initial diagnosis. All electrophoresis-defined urinary Bence-Jones proteins were labeled by the anti-free λ light chain (anti-fλ) reagent; none were labeled by the anti-free κ light chain (anti-fκ); 2 of these were identified as fκ by mass spectrometry. An electrophoretically identical protein restriction that was labeled by the anti-fλ reagent was present in the paired serum from 5/8 of cases, documenting sfLC. Conclusions and clinical importance Commercially available human IF reagents identified sfLC and ufLC in both dogs and cats. Free light chains may be nephrotoxic in dogs.

With the first case of COVID-19 confirmed in Canada in early 2020, our country joined in the fight against a novel pathogen in a global pandemic. The stress of uncertainty and practice change was most apparent in the emergency department when it came to managing known or suspected COVID-19 patients requiring airway management. Recognizing the need for a coordinated approach, a province wide rapid response distributed model of continuing professional development for airway management was developed utilizing Airway Leads to help prepare front-line medical personnel providing airway management for these patients. Airway Leads worked with local physicians to deliver consistent, high quality airway education across the province during the initial surge of cases. Education included both in person and virtual sessions along with real time ongoing support through provincial guidelines, videos, and other documents. Physician reported "stress level" pre- and post-Airway Lead support declined from a median score of 9 to 7 (on a 10-point Likert Scale).RéSUMé: Le premier cas de COVID-19 ayant été confirmé au Canada au début de 2020, notre pays s’est joint à la lutte contre un nouveau pathogène dans une pandémie mondiale. Le stress de l’incertitude et du changement de pratique était plus évident au service d’urgence lorsqu’il s’agissait de gérer les patients connus ou soupçonnés de la COVID-19 qui avaient besoin d’une prise en charge des voies respiratoires. Reconnaissant la nécessité d'une approche coordonnée, un modèle de développement professionnel continu distribué à l'échelle de la province pour la gestion des voies aériennes a été élaboré en utilisant les Airway Leads pour aider à préparer le personnel médical de première ligne qui assure la gestion des voies aériennes de ces patients. Airway Leads a travaillé avec les médecins locaux pour dispenser un enseignement cohérent et de haute qualité sur les voies aériennes dans toute la province lors de l'augmentation initiale du nombre de cas. L'éducation comprenait à la fois des sessions en personne et virtuelles ainsi qu'un soutien continu en temps réel par le biais de directives provinciales, de vidéos et d'autres documents. Le "niveau de stress" déclaré par le médecin avant et après l'intervention de Airway Lead a diminué, passant d'un score médian de 9 à 7 (sur une échelle de Likert de 10 points).

Background Current recommendations for monitoring disease progression and response to treatment in humans with multiple myeloma include evaluation of serum paraprotein (M-protein) concentration. Densitometry, species-specific radial immunodiffusion (RID) and ELISA methods can be used to quantify M-proteins. Objective Retrospectively evaluate use of the International Myeloma Working Group (IMWG) response criteria for humans in dogs with multiple myeloma. Animals Sixteen dogs with a diagnosis of multiple myeloma, M-protein documented by serum protein electrophoresis (SPE) and immunofixation (IF) in an initial sample and subsequent electrophoretic evaluation of serial samples. Methods Retrospectively, densitometric M-proteins, RID and globulins were measured and characterized according to IMWG criteria. Available clinical history was reviewed. Overall survival time (OST) was calculated from initial electrophoretic evaluation to death or last contact. Results All cases received some form of nonstandardized chemotherapy. Complete response (CR), a lack of detectable M-protein by SPE and IF, was documented in 1 case. Median survival was longer for dogs that attained ≥90% densitometric M-protein reduction (630 days) than for those that did not attain at least 50% reduction in densitometric M-protein (284 days; log rank P = .006). Five dogs were defined as having progressive disease (M-protein increase of >25% and at least 0.5 g/dL from nadir), which correlated with concurrent or subsequent clinical deterioration. Response criteria categorized by serum globulins or RID was not correlated with OST or clinical findings. Conclusions and clinical importance Densitometric M-protein characterized using IMWG response criteria correlated with OST and clinical findings. Densitometric M-protein detection should be used to monitor dogs with multiple myeloma.

Hyperglobulinemia is reported in 26% of canine chronic B-cell lymphocytic leukemia (B-CLL) cases. However, few cases have been characterized by protein electrophoresis and immunofixation (IF), and the incidence of a monoclonal protein (M-protein) is unknown using these techniques.To characterize and determine the proportion of canine B-CLL cases with an M-protein using plasma protein electrophoresis (PPE), routine and free light chain (fLC) IF, and to assess if productive B-CLL cases express MUM1/IRF4 by cell tube block (CTB).PPE, routine (targeting IgG, IgA, IgM, IgG4, and light chain) and fLC IF were performed using 48 dog B-CLL plasma samples from patients diagnosed via peripheral blood flow cytometry. CTB was performed on a separate cohort of 15 patients.Hyperproteinemia (7.5 g/dL) was present in 17/48 cases (35%). An M-protein was detected in 32/48 cases (67%). Of these, 19/32 cases (59%) had only complete (monoclonal heavy and light chain) M-proteins detected, 10/32 cases (31%) had both complete and fLC M-proteins detected, and 3/32 cases (9%) had only an fLC M-protein detected. IgM was the most common clonal immunoglobulin isotype detected (23 cases). CD21Most B-CLL cases had an M-protein and were not hyperproteinemic. Most cases with paraproteins had a complete IgM monoclonal gammopathy; a subset had documented fLCs. The prognostic significance of heavy and fLC presence should be evaluated.

Three dogs under 12 months old were diagnosed with atypical multiple myeloma (MM), having an aggressive multifocal anaplastic round cell sarcoma in bone marrow, viscera, and/or peripheral blood, which were confirmed by cytology and immunohistochemistry to be of plasma cell origin. The intramedullary sarcomas caused myelophthisis, osteolysis, and hypercalcemia. Complete or free light chain monoclonal gammopathy in the serum and/or urine was demonstrated by protein electrophoresis and immunofixation. The polymerase chain reaction for antigen receptor rearrangement assay performed on 2 cases identified a clonally rearranged immunoglobulin gene. Neoplastic cells lacked expression of CD45, CD3, CD18, CD21, CD34, and MHCII by flow cytometry. Immunohistochemistry revealed MUM1 immunoreactivity of the neoplastic cells. Combining all data, the diagnosis was MM. An aggressive form of MM in young dogs should be a differential diagnosis for patients with an immunoglobulin-productive, B cell-clonal, CD45-negative, MUM1-positive discrete cell neoplasm arising from the bone marrow.

Colorectal cancer (CRC) is the second most prevelant malignancy in Europe and diet is an important modifiable risk factor. Processed meat consumption, including meats with preservative salts such as sodium nitrite, have been implicated in CRC pathogenesis. This study investigated how the CRC pathology and metabolic status of adenomatous polyposis coli (APC) multiple intestinal neoplasia (min) mice was perturbed following 8 weeks of pork meat consumption. Dietary inclusions (15%) of either nitrite-free pork, nitrite-free sausage, or nitrite-containing sausage (frankfurter) were compared against a parallel control group (100% chow). Comprehensive studies investigated: gastrointestinal tract histology (tumours), aberrant crypt foci (ACF), mucin deplin foci (MDF), lipid peroxidation (urine and serum), faecal microbiota, and serum metabolomics (599 metabolites). After 8 weeks mice consuming the frankfurter diet had 53% more (P = 0.014) gastrointestinal tumours than control, although ACF and MDF did not differ. Urine and serum lipid peroxidation markers were 59% (P = 0.001) and 108% (P = 0.001) higher, respectively in the frankfurter group. Gut dysbiosis was evident in these mice with comparably fewer Bacteriodes and more Firmicutes. Fasting serum levels of trimethylamine N-oxide (TMAO) and numerous triglycerides were elevated. Various serum phosphotidylcholine species were decreased. These results demonstrate that nitrite-containing sausages may exaccerbate the development of CRC pathology in APC

Colorectal cancer (CRC) is the second most prevelant malignancy in Europe and diet is an important modifiable risk factor. Processed meat consumption, including meats with preservative salts such as sodium nitrite, have been implicated in CRC pathogenesis. This study investigated how the CRC pathology and metabolic status of adenomatous polyposis coli (APC) multiple intestinal neoplasia(min) mice was perturbed following 8 weeks of pork meat consumption. Dietary inclusions (15%) of either nitrite-free pork, nitrite-free sausage or nitrite-containing sausage (frankfurter) were compared against a parallel control group (100% chow). Comprehensive studies investigated: gastrointestinal tract histology (tumours, aberrant crypt foci (ACF) and mucin deplin foci (MDF), lipid peroxidation (urine and serum), faecal microbiota and serum metabolomics (599 metabolites). After 8 weeks mice consuming the frankfurter diet had 53% more (P=0.014) gastrointestinal tumours than control, although ACF and MDF did not differ. Urine and serum lipid peroxidation markers were 59% (P=0.001) and 108% (P=0.001) higher, respectively in the frankfurter group. Gut dysbiosis was evident in these mice with comparably fewerBacteriodesand moreFirmicutes. Fasting serum levels of trimethylamine N-oxide (TMAO) and numerous triglycerides were elevated. Various serum phosphotidylcholine species were decreased. These results demonstrate that nitrite-containing sausages may exaccerbate the development of CRC pathology in APCMinmice to a greater extent than nitrite-free sausages, and this is associated with greater lipid peroxidation, wide-ranging metabolic alternation and gut dysbiosis.

A 12-y-old spayed female Schipperke dog with a previous diagnosis of inflammatory bowel disease was presented with a 2-mo history of severe colitis. The patient’s condition progressed to hepatopathy, pneumonia, and dermatitis following management with prednisolone and dexamethasone sodium phosphate. Colonic biopsies identified severe necrosuppurative colitis with free and intracellular parasitic zoites. Postmortem examination confirmed extensive chronic-active ulcerative colitis, severe acute necrotizing hepatitis and splenitis, interstitial pneumonia, ulcerative dermatitis, myelitis (bone marrow), and mild meningoencephalitis with variable numbers of intracellular and extracellular protozoal zoites. PCR on samples of fresh colon was positive for Neospora caninum. Immunohistochemistry identified N. caninum tachyzoites in sections of colon, and a single tissue cyst in sections of brain. Administration of immunosuppressive drugs may have allowed systemic dissemination of Neospora from the intestinal tract.

Objective Neuronal ceroid lipofuscinosis type 2 (CLN2 disease) is a rare, progressive, fatal neurodegenerative pediatric disorder resulting from deficiencies of the lysosomal enzyme tripeptidyl peptidase 1 that are caused by mutations in TPP1. Identifying biomarkers of CLN2 disease progression will be important in assessing the efficacy of therapeutic interventions for this disorder. Neurofilament light is an intrinsic component of healthy neurons; elevated circulating extracellular neurofilament light is a biomarker of neuropathology in several adult‐onset neurological diseases. Our objective was to assess whether circulating neurofilament light is a biomarker that is responsive to enzyme replacement therapy (ERT) in CLN2 disease. Methods Using an ultrasensitive immunoassay, we assessed plasma neurofilament light changes during disease progression in a canine model of CLN2 disease and in ERT clinical trial CLN2 disease patients. Results In tripeptidyl peptidase 1 (TPP1)‐null dogs (N = 11), but not in control dogs [N = 6 (TPP1 +/−) and N = 27 (WT)], neurofilament light levels increased more than tenfold above initial low baseline levels during disease progression. Before treatment in 21 human subjects with CLN2 disease (age range: 1.72–6.85 years), neurofilament light levels were 48‐fold higher (P

An emerging theory about racial differences in cancer risk and outcomes is that psychological and social stressors influence cellular stress responses, however, limited empirical data are available on racial differences in cellular stress responses among men who are at risk for adverse prostate cancer outcomes. In this study, we undertook a systems approach to examine molecular profiles and cellular stress responses in an important segment of African American (AA) and European American (EA) men: men undergoing prostate biopsy. We assessed the prostate transcriptome with a single biopsy core via high throughput RNA sequencing (RNA-Seq). Transcriptomic analyses uncovered impacted biological pathways including PI3K-Akt signaling pathway, Neuroactive ligand-receptor interaction pathway, and ECM-receptor interaction. Additionally, 187 genes mapping to the Gene Ontology (GO) terms RNA binding, structural constituent of ribosome, SRP-dependent co-translational protein targeting to membrane and the biological pathways, translation, L13a-mediated translational silencing of Ceruloplasmin expression were differentially expressed (DE) between EA and AA. This signature allowed separation of AA and EA patients, and AA patients with the most severe clinical characteristics. AA patients with elevated expression levels of this genomic signature presented with higher Gleason scores, a greater number of positive core biopsies, elevated dehydroepiandrosterone sulfate levels and serum vitamin D deficiency. Protein-protein interaction (PPI) network analysis revealed a high degree of connectivity between these 187 proteins.

Achilles’ heel relationships arise when the status of one gene exposes a cell's vulnerability to perturbation of a second gene, such as chemical inhibition, providing therapeutic opportunities for precision oncology. SynLeGG (www.overton-lab.uk/synlegg) identifies and visualizes mutually exclusive loss signatures in ‘omics data to enable discovery of genetic dependency relationships (GDRs) across 783 cancer cell lines and 30 tissues. While there is significant focus on genetic approaches, transcriptome data has advantages for investigation of GDRs and remains relatively underexplored. SynLeGG depends upon the MultiSEp algorithm for unsupervised assignment of cell lines into gene expression clusters, which provide the basis for analysis of CRISPR scores and mutational status in order to propose candidate GDRs. Benchmarking against SynLethDB demonstrates favourable performance for MultiSEp against competing approaches, finding significantly higher area under the Receiver Operator Characteristic curve and between 2.8-fold to 8.5-fold greater coverage. In addition to pan-cancer analysis, SynLeGG offers investigation of tissue-specific GDRs and recovers established relationships, including synthetic lethality for SMARCA2 with SMARCA4. Proteomics, Gene Ontology, protein-protein interactions and paralogue information are provided to assist interpretation and candidate drug target prioritization. SynLeGG predictions are significantly enriched in dependencies validated by a recently published CRISPR screen., Graphical Abstract Graphical AbstractSynLeGG utilizes MultiSEp gene expression clusters to partition CRISPR essentiality scores (top) and mutations from whole exome sequencing (bottom), in order to predict candidate genetic dependency relationships, including synthetic lethality.

Ipatasertib is a highly selective small-molecule pan-Akt inhibitor in clinical development. Ipatasertib is predominantly eliminated by the liver, and therefore the effect of hepatic impairment on ipatasertib pharmacokinetics (PK) was evaluated. In this Phase I open-label, parallel group study, the PK of ipatasertib was evaluated in subjects with hepatic impairment based on both, the Child-Pugh and the National Cancer Institute-Organ Dysfunction Working Group (NCI-ODWG) classification for hepatic impairment. A single dose of ipatasertib at 100 mg was administered and the PK was characterized in healthy subjects with normal hepatic function or mild, moderate and severe hepatic impairment. Based on Child-Pugh classification, subjects with moderate or severe hepatic impairment had an approximately 2- and 3-fold increase in systemic exposure (AUC0-∞ ) to ipatasertib, respectively, compared to subjects with normal hepatic function. Systemic exposure (AUC0-∞ ) to ipatasertib in subjects with mild hepatic impairment was comparable to that in subjects with normal hepatic function. In accordance with reduced clearance capacity, subjects with mild to severe hepatic impairment showed lower systemic exposure (AUC0-∞ ) of ipatasertib metabolite M1 (G-037720). Overall results were comparable between Child-Pugh and NCI-ODWG classification criteria. Based upon the results from this study, no dosage adjustment is required for ipatasertib when treating patients with mild hepatic impairment, whereas a dose reduction would be recommended for subjects with moderate or severe hepatic impairment. Based on real world data analysis, ∼2% of intended patient population is expected to need a modified dose due to moderate or severe hepatic impairment. This article is protected by copyright. All rights reserved.

Perkembangan teknologi telah memasuki tahap dimana penggunaan aplikasi atau perangkat lunak sebagai core dari sebuah bisnis. Hal ini dilihat dari munculnya berbagai sistem informasi yang mulai dijadikan aset penting dalam mengembangkan sebuah bisnis usaha, salah satunya dalam bidang akademik. Seiring dengan bertambah dan berkembangnya kebutuhan pengguna maka sistem/aplikasi yang digunakan akan semakin membesar. Melalui tahapan testing kualitas sistem yang digunakan akan dapat terjaga dan terdokumentasi. Ma Chung Integrated Information System (macIS) merupakan sistem terintegrasi yang digunakan universitas Ma Chung sebagai pengolah data-data dan informasi akademik/non akademik mahasiswa, dosen, dan staff. Hingga saat ini macIS masih belum memiliki dokumentasi testing, maka dari itu diperlukan dokumentasi yang terstandar agar kualitas sistem informasi yang dimiliki universitas dapat terjaga dan berkembang dengan baik.Berdasarkan latar belakang, terdapat dua masalah utama dalam sistem macIS. Pertama, macIS belum memiliki dokumentasi testing yang menyebabkan kualitas sistem tidak dapat dikontrol dengan baik. Kedua, tidak ada pendokumentasian yang terstandar pada sistem informasi universitas. Sedangkan Pengujian sistem dilakukan dengan menggunakan metodologi STLC(Software Testing Life Cycle) yang membagi proses pengujian sistem menjadi enam bagian mulai dari fase analisis, perencanaan, desain, spesifikasi kebutuhan perangkat, eksekusi, hingga ditutup dengan hasil luaran berupa dokumentasi pengujian perangkat lunak. Hasil akhir yang didapat adalah dokumentasi berupa skenario uji berdasarkan 4 aspek pada sistem dan lampiran dokumen MTP(Master Test Plan) berstandar IEEE-829.

Densitometric quantitation using serum protein electrophoresis (SPE) is used to monitor monoclonal proteins (M-proteins) in human patients but has not been validated in the dog. Serum globulin concentrations, species-specific radial immunodiffusion (RID), and ELISAs are currently used in veterinary medicine.We aimed to compare four methods that quantify M-proteins using densitometry and biuret protein (dM-protein) measurements. We also validated the best performing method and compared it with the RID and ELISA methods for measuring canine serum M-protein.Serum from six normal dogs and 83 serum samples from 46 dogs with confirmed monoclonal gammopathies were used. A spike and recovery experiment with purified monoclonal IgG and IgM, inter-run and intra-run variability, linearity under dilution, and lower limit of detection were performed. Results of commercial canine RID and ELISA kits for total class-specific immunoglobulin were compared with dM-proteins.The corrected perpendicular drop gating method had20% error for IgG/γ-globulin and IgM/β-globulin M-protein quantifications. Linearity (r .99), intra-run CV (1.1%-2.3%), and inter-run CVs (2.0%-3.5%) were acceptable. Correlation between the RID and densitometry results ranged from r = .25 to r = .88, depending on the class. The RID result was greater than that of the biuret total protein in 26/63 (41%) IgA cases. A panel of IgG, IgA, and IgM RIDs failed to correctly identify an IgM paraproteinemia in 6/6 (100%) cases. Densitometry was not comparable with any other tested method.Densitometric quantitation is a valid technique for measuring M-proteins in the β- and γ-globulin regions. Immunotyping via RID using the tested kit does not appear to detect IgM. Densitometry is recommended for measuring M-proteins in canine patients.

Summary Flexible navigation relies on a cognitive map of space, thought to be implemented by hippocampal place cells: neurons that exhibit location-specific firing. In connected environments, optimal navigation requires keeping track of one’s location and of the available connections between subspaces. We examined whether the dorsal CA1 place cells of rats encode environmental connectivity in four geometrically identical boxes arranged in a square. Rats moved between boxes by pushing saloon-type doors that could be locked in one or both directions. Although rats demonstrated knowledge of environmental connectivity, their place cells did not respond to connectivity changes, nor did they represent doorways differently from other locations. Place cells coded location in a global reference frame, with a different map for each box and minimal repetitive fields despite the repetitive geometry. These results suggest that CA1 place cells provide a spatial map that does not explicitly include connectivity., Highlights • Rats flexibly navigate in a four-room maze where connectivity changes • Place cell firing fields are not specifically altered by changes in connectivity • Single or ensemble place cell activity does not encode changes in connectivity • Place cells can uniquely map identical connected compartments, Hippocampal place cells are thought to underlie a “cognitive map” of space. Duvelle and Grieves et al. show that the place cells of rats discriminate four connected rooms and rats learn changes in their connectivity. However, such changes do not affect place cell activity, suggesting that other brain regions or mechanisms encode this information.

95 Background: mCRPC remains an incurable disease with unmet need for improved therapies. PARP inhibition has demonstrated antitumor activity, and preclinical studies suggest synergy between PARP and AKT inhibition. This phase Ib trial (NCT03840200) sought to determine the maximum tolerated dose of PARP inhibitor rucaparib (ruca) with AKT inhibitor ipatasertib (ipat) and explore early safety and efficacy in patients (pts) with mCRPC. Methods: The study had 2 parts: a part 1 dose-escalation phase (cohort [C]1: ipat 300 mg QD + ruca 400 mg BID; C2a: ipat 300 mg QD + ruca 600 mg BID; C2b ipat 400 mg QD + ruca 400 mg BID; C3: ipat 400 mg QD + ruca 600 mg BID) in unselected pts with mCRPC or advanced breast or ovarian cancer, and a Part 2 dose-expansion phase in pts with mCRPC whose previous androgen deprivation therapy failed. The primary safety endpoints were AEs and dose-limiting toxicities, with the goal of identifying a recommended phase II dose (RP2D). The primary efficacy endpoint was prostate-specific antigen (PSA) response (PSA reduction of ≥ 50%) in pts with mCRPC. Secondary efficacy endpoints included ORR per RECIST 1.1, radiographic PFS (rPFS) per Prostate Cancer Working Group 3 and OS in pts with mCRPC. We assessed homologous recombination deficiency (HRD), PIK3CA/ AKT1/ PTEN status and other alterations by next-generation sequencing. Results: Part 1 enrolled 21 pts: 17 with mCRPC, 3 with ovarian cancer and 1 with breast cancer (C1 [n=8], C2a [n=6] and C2b [n=7], C3 [not opened]). Ipat 400 mg QD + ruca 400 mg BID was the selected RP2D. 30 pts enrolled in Part 2, so 37 pts with mCRPC (C2b + Part 2) received the RP2D (median follow-up 93 days). All grade AEs occurred in 37 pts (100%), serious AEs in 8 pts (22%) and AEs leading to treatment modification in 26 pts (70%). The most frequent AEs (all grade and grade 3, respectively) were diarrhea (35 [95%] and 2 [5%]); asthenia (24 [65%] and 4 [11%]); and nausea (24 [65%] and 2 [5%]). There was 1 grade 4 AE (anemia) and no grade 5 AEs. See the table for efficacy data. In biomarker subgroups, the PSA response rate was 50% (3/6) in pts with HRD tumors vs 25% (4/16) in pts with HR-proficient tumors, and 14% (1/7) vs 40% (6/15) in pts with PIK3CA/ AKT1/ PTEN–altered or intact tumors, respectively. Conclusions: The combination of ipat and ruca was well tolerated but did not show clinically relevant antitumor activity in pts with mCRPC, including in predefined biomarker subpopulations. Clinical trial information: NCT03840200. [Table: see text]

Flexible navigation relies on a cognitive map of space, thought to be implemented by hippocampal place cells: neurons that exhibit location-specific firing. In connected environments, optimal navigation requires keeping track of one’s location and of the available connections between subspaces. We examined whether the dorsal CA1 place cells of rats encode environmental connectivity in four geometrically-identical boxes arranged in a square. Rats moved between boxes by pushing saloon-type doors that could be locked in one or both directions. While rats demonstrated knowledge of environmental connectivity, their place cells did not respond to connectivity changes, nor did they represent doorways differently from other locations. Importantly, place cells coded the space in a global frame, expressing minimal repetitive fields despite the repetitive geometry (global coding). These results suggest that CA1 place cells provide a spatial map that does not explicitly include connectivity.

Background The diagnostic performance of routine electrophoresis (agarose gel electrophoresis [AGE] and capillary zone electrophoresis [CZE]) and species-specific immunofixation (IF) for the detection of immunoglobulin paraproteins (M-proteins) and diagnosis of secretory myeloma-related disorders (sMRD) can be improved. Available canine IF targets were IgG-FC, IgA, IgM, light chain (LC), IgG4, and free LC (fLC) antibodies. Objective We aimed to review specific features associated with the presence of M-proteins in canine serum samples and the common features causing inaccurate reporting of M-proteins to improve the diagnostic performance of routine electrophoresis and IF for the detection of M-proteins. Methods Features found in AGE, CZE, routine IF, IgG4 IF, and fLC IF of 100 canine serum samples from Part 1 of this study were evaluated by simple and multivariate logistic regression to identify factors associated with the presence of M-proteins. Cases falsely called negative or positive for M-proteins were reviewed to identify the common features that could be used to increase the diagnostic performance of SPE and IF for M-protein detection. Results The presence of hypogammaglobulinemia or any peak taller than albumin was associated with an M-protein. Total protein concentrations, globulin concentrations, or peaks wider than albumin were not associated with an M-protein. Free LC sMRD cases were not diagnosed by SPE and routine IF. Cases with infectious and inflammatory etiologies had a restricted polyclonal gammopathy with multiple γ-globulin restrictions resulting in some false-positive results. SPE combined with all available IF results and the specific features identified in this study had an estimated sensitivity of 95.1% and specificity of 81.4%. Conclusions The identified criteria of this study increase the diagnostic performance of the electrophoretic evaluation for M-proteins.

A 12-y-old spayed female Schipperke dog with a previous diagnosis of inflammatory bowel disease was presented with a 2-mo history of severe colitis. The patient’s condition progressed to hepatopathy, pneumonia, and dermatitis following management with prednisolone and dexamethasone sodium phosphate. Colonic biopsies identified severe necrosuppurative colitis with free and intracellular parasitic zoites. Postmortem examination confirmed extensive chronic-active ulcerative colitis, severe acute necrotizing hepatitis and splenitis, interstitial pneumonia, ulcerative dermatitis, myelitis (bone marrow), and mild meningoencephalitis with variable numbers of intracellular and extracellular protozoal zoites. PCR on samples of fresh colon was positive for Neospora caninum. Immunohistochemistry identified N. caninum tachyzoites in sections of colon, and a single tissue cyst in sections of brain. Administration of immunosuppressive drugs may have allowed systemic dissemination of Neospora from the intestinal tract.

Aim: To determine whether platelet-rich plasma (PRP) can successfully treat symptoms associated with proximal hamstring tendinopathy. Materials & methods: Charts were retrospectively reviewed of patients with a diagnosis of chronic proximal hamstring tendinopathy who underwent an ultrasound-guided leukocyte-rich PRP injection to assess reported outcomes at baseline and final follow-up post-intervention. Results: In 22 patients with a mean age of 48.7 years and mean symptom duration of 26.5 months, mean Numeric Pain Rating Scale and Victorian Institute of Sport Assessment Scale for Proximal Hamstring Tendinopathy subscores demonstrated statistically significant improvements (p < 0.05) at a mean of 7.91 months follow-up. Sixty-eight percent of patients demonstrated ≥50% reduction of pain. Conclusion: Pain and functional outcomes improved following injection of PRP.

Flexible navigation relies on a cognitive map of space, thought to be implemented by hippocampal place cells: neurons that exhibit location-specific firing. In connected environments, optimal navigation requires keeping track of one's location and of the available connections between subspaces. We examined whether the dorsal CA1 place cells of rats encode environmental connectivity in four geometrically-identical boxes arranged in a square. Rats moved between boxes by pushing saloon-type doors that could be locked in one or both directions. While rats demonstrated knowledge of environmental connectivity, their place cells did not respond to connectivity changes, nor did they represent doorways differently from other locations. Importantly, place cells coded the space in a global frame, expressing minimal repetitive fields despite the repetitive geometry (global coding). These results suggest that CA1 place cells provide a spatial map that does not explicitly include connectivity.

Bullous amyloidosis is a rare disease in humans that has not been described in a veterinary species in the peer-reviewed literature. The human disease is characterised by haemorrhagic vesicles and bullae on the skin and mucosae, which form due to amyloid deposition.To describe the clinical features, laboratory analysis and histopathological features of an unique presentation of bullous disease in a horse.A 17-year-old thoroughbred mare presented for weight loss and severe oral cavity ulcers.Investigations involved haematological evaluation, chemistry profiles, gastroscopy and serum protein electrophoresis, and, postmortem, histopathological evaluation, Congo-red staining and transmission electron microscopy (TEM).Haemorrhagic vesicles and bullae occurred on the mucosa of the oral cavity, lips, oesophagus and stomach, and much less the muzzle, face and mucocutaneous areas of the perineum, where scarring was evident. Histopathological evaluation and Congo-red staining confirmed the presence of amyloid deposits in dermis and submucosa, in association with vesicle and bulla formation, consistent with bullous amyloidosis. TEM confirmed amyloid fibril deposition in the dermis and along the basement membrane zone. Clefts occurred in the superficial dermis and submucosa, which explained haemorrhage and scarring. The presence of a polyclonal gammopathy and the rapid abolishment of Congo-red staining with performate pretreatment supported serum amyloid A and secondary amyloidosis.Bullous amyloidosis is a novel disease of the horse and a newly recognised differential for bullous disease, for which the haemorrhagic nature of bullae, scarring and deep secondary ulcers are considered clinical clues to the condition.L’amyloïdose bulleuse est une maladie rare chez l’homme qui n’a pas encore été décrite dans une espèce vétérinaire dans la littérature avec comité de lecture. La maladie de l’homme est caractérisée par des vésicules et bulles hémorragiques de la peau et des muqueuses qui se forment après dépôt d’amyloïde. HYPOTHÈSES/OBJECTIFS: Décrire les données cliniques, les analyses de laboratoires et les critères histopathologiques d’une présentation unique de maladie bulleuse chez un cheval.Une jument de race de 17 ans présentée pour perte de poids et ulcères sévères de la cavité buccale. MÉTHODES: Les investigations comprenaient une évaluation hématologique, des profils biochimiques, une gastroscopie et une électrophorèse des protéines sériques et, post mortem, un examen histopathologique, une coloration au rouge Congo et une microscopie à transmission électronique (TEM). RÉSULTATS: Les vésicules et bulles hémorragiques se sont développées sur les muqueuses de la cavité orale, les lèvres, l’œsophage et l’estomac et moins au niveau des narines, de la face et des jonctions cutanéo-muqueuses du périnée où des cicatrices étaient évidentes. Une évaluation histopathologique et une coloration au rouge Congo ont confirmé la présence d’amyloïde dans le derme et les muqueuses, en association avec la formation des vésicules et des bulles, compatible avec l’amyloïdose bulleuse. La TEM a confirmé le dépôt de fibrilles d’amyloïdes dans le derme et au niveau de la membrane basale. Un clivage était présent dans le derme superficiel et les sous muqueuses, expliquant l’hémorragie et les cicatrices. La présence d’une gammapathie polyconale et du rapide disparition de la coloration au rouge Congo avec réalisation d’un prétraitement, soutenaient une amyloïde A sérique et une amyloïdose secondaire.L’amyloïdose bulleuse est une nouvelle maladie du cheval et un nouveau différentiel dans les dermatoses bulleuses pour laquelle la nature hémorragique des bulles, des cicatrices et des ulcères secondaires profonds est considérée comme une donnée clinique évocatrice de l’atteinte.INTRODUCCIÓN: la amiloidosis bullosa es una enfermedad rara en humanos que no se ha descrito en ninguna especie veterinaria en la literatura revisada. La enfermedad humana se caracteriza por vesículas y ampollas hemorrágicas en la piel y las mucosas, que se forman debido al depósito de amiloide. HIPÓTESIS/OBJETIVOS: describir las características clínicas, análisis de laboratorio y características histopatológicas de una presentación única de enfermedad bullosa en un caballo. ANIMALES: una yegua pura sangre de 17 años que se presentó por pérdida de peso y úlceras severas en la cavidad oral. MÉTODOS: Las investigaciones incluyeron evaluación hematológica, perfiles químicos, gastroscopia y electroforesis de proteínas séricas y, post mortem, evaluación histopatológica, tinción con rojo Congo y microscopía electrónica de transmisión (TEM). RESULTADOS: Se observaron vesículas y ampollas hemorrágicas en la mucosa de la cavidad oral, labios, esófago y estómago, y mucho menos en el hocico, la cara y las áreas mucocutáneas del perineo, donde las cicatrices eran evidentes. La evaluación histopatológica y la tinción con rojo Congo confirmaron la presencia de depósitos de amiloide en dermis y submucosa, en asociación con formación de vesículas y ampollas, compatibles con amiloidosis bullosa. TEM confirmó el depósito de fibrillas de amiloide en la dermis y a lo largo de la zona de la membrana basal. Se produjeron hendiduras en la dermis superficial y la submucosa, lo que explica la hemorragia y la cicatrización. La presencia de una gammapatía policlonal y la rápida desaparición de la tinción con rojo Congo con el pretratamiento con permanganato potásico confirmaron amiloide A sérico y la amiloidosis secundaria. CONCLUSIÓN E IMPORTANCIA CLÍNICA: la amiloidosis bullosa es una enfermedad nueva del caballo y un diferencial recientemente reconocido de enfermedades bullosas. La naturaleza hemorrágica de las ampollas, la presencia de cicatrices y de úlceras secundarias profundas se consideran indicios clínicos de esta enfermedad.Die bullöse Amyloidose ist eine seltene Erkrankung beim Menschen, die noch bei keiner tierischen Spezies in Peer-Review Literatur beschrieben wurde. Die Erkrankung ist beim Menschen durch hämorrhagische Bläschen und Blasen auf der Haut und den Schleimhäuten gekennzeichnet, die sich aufgrund von Amyloid Ablagerung bilden.Die Beschreibung der klinischen Merkmale, der Laboranalyse und der histopathologischen Merkmale einer einzigartigen Präsentation einer bullösen Erkrankung bei einem Pferd.Eine 17 Jahre alte Vollblutaraberstute wurde wegen Gewichtsverlust und hochgradigen Ulzera in der Mundhöhle vorgestellt.Die Untersuchungen beinhalteten Hämatologie, Biochemie, Gastroskopie und eine Serumelektrophorese, sowie post mortem eine histopathologische Evaluierung, Kongorotfärbung und Transmissionselektronenmikroskopie (TEM).Hämorrhagische Bläschen und Blasen traten an der Mundschleimhaut, an den Lippen, der Speiseröhre und im Magen auf, sowie etwas weniger ausgeprägt an der Nase, dem Gesicht und den mucokutanen Übergängen des Perneums, wo Narbenbildung evident war. Die histopathologische Evaluierung und die Kongorotfärbung bestätigten das Vorkommen von Amyloidablagerungen in der Dermis und Submukosa, im Zusammenhang mit Bläschen- und Blasenbildung, was mit einer bullösen Amyloidose übereinstimmte. Die TEM bestätigte Amyloidfasernablagerungen in der Dermis und entlang der Basalmembranzone. In der oberflächlichen Dermis und Submukosa bestand eine Spaltenbildung, die die Blutung und die Narbenbildung erklärten. Das Auftreten eine polyklonalen Gammopathie sowie das rasche Verschwinden der Kongorotfärbung nach performativer Vorbehandlung stützte die Diagnose einer Serum Amyloid A und einer sekundären Amyloidose.Die bullöse Amyloidose ist eine neue Erkrankung des Pferdes und eine neu erkannte Differentialdiagnose für bullöse Erkrankungen, bei der die hämorrhagische Natur der Blasen, die Narbenbildung und die tiefen sekundären Ulzera als klinische Hinweise für diesen Zustand gelten.背景: 水疱性アミロイドーシスは、ヒトではまれな疾患で、獣医学的にはまだ報告されていない。本疾患は、アミロイド沈着により皮膚や粘膜に形成される出血性の小水疱や水疱が特徴である。 仮説/目的: 本研究の目的は、1頭の馬に認められた水疱疾患のユニークな症状の臨床的特徴、実験室解析および病理組織学的特徴を説明することであった。 供試動物: 17歳のサラブレッドの雌馬が体重減少および重度の口腔内潰瘍のために来院した。 方法: 血液学的評価、化学的プロファイル、胃内視鏡検査、血清タンパク質電気泳動を行い、死後、組織学的評価、コンゴレッド染色、透過型電子顕微鏡 (TEM) を行った。 結果: 出血性小水疱および水疱は口腔、口唇、食道、胃の粘膜に発生し、鼻口部、顔面、会陰粘膜皮膚領域にはほとんど発生せず、瘢痕が認められた。病理組織学的評価およびコンゴレッド染色により、水疱性アミロイドーシスと一致した、小水疱や水疱の形成と関連する真皮および粘膜下層のアミロイド沈着の存在が確認された。TEMでは、真皮および基底膜帯に沿ってアミロイド線維の沈着が確認された。表層の真皮および粘膜下層に裂け目が生じており, 出血および瘢痕化が説明できた。ポリクローナルガンモパシーの存在と、前治療の実施によるコンゴレッド染色の急速な廃絶は、血清アミロイド A および続発性アミロイドーシスを支持した。 結論と臨床的重要性: 水疱性アミロイドーシスは馬の新しい病気であり、水疱症の鑑別として新たに認識された。水疱の出血性の性質、瘢痕、深い二次的な潰瘍がこの病気の臨床的な手がかりと考えられる。.背景: 大疱性淀粉样变性是一种人类罕见疾病, 动物发病尚未在兽医同行评审文献中描述过。人类疾病的特征是皮肤和粘膜上的出血性水疱和大疱, 其由淀粉样蛋白沉积形成。 假设/目的: 描述马大疱性疾病独特表现的临床特征、实验室分析和组织病理学特征。 动物: 只17岁的纯种母马因体重减轻和严重的口腔溃疡而就诊。 方法: 研究涉及血液学评价、化学特征、胃镜检查和血清蛋白电泳, 以及尸检、组织病理学评价、刚果红染色和透射电子显微镜(TEM)。 结果: 口腔、嘴唇、食管和胃的粘膜上发生出血性水疱和大疱, 更少发生在有明显瘢痕的口鼻、面部和会阴皮肤粘膜区域。组织病理学评价和刚果红染色证实真皮和粘膜下层存在淀粉样沉积物, 与水泡和大疱形成相关, 与大疱性淀粉样变性一致。TEM证实淀粉样纤维沉积于真皮, 并沿基底膜区沉积。裂隙发生在真皮浅层和粘膜下层, 这解释了出血和瘢痕形成。多克隆丙种球蛋白病的存在和进行预处理后刚果红染色的快速消除,支持了血清淀粉样蛋白a和继发性淀粉样变性。 结论和临床意义: 大疱性淀粉样变性是马的一种新型疾病, 并发现与大疱性疾病存在差异, 大疱的出血性质、瘢痕和深部继发性溃疡被认为是该疾病的临床线索。.A amiloidose bolhosa é uma doença rara em humanos que não foi descrita em espécies veterinárias na literatura com sistema de revisão por pares. A doença humana é caracterizada por vesículas e bolhas hemorrágicas na pele e mucosas, que se formam devido à deposição de substância amiloide. HIPÓTESE/OBJETIVOS: Descrever as características clínicas, exames laboratoriais e histopatológicos de uma doença bolhosa com apresentação singular em um cavalo.Uma égua da raça puro-sangue inglês de 17 anos de idade foi apresentada devido a um quadro de perda de peso e úlceras graves da cavidade oral. MÉTODOS: As investigações consistiram de avaliação hematológica, perfil bioquímico, gastroscopia e eletroforese de proteínas séricas e, postmortem, avaliação histopatológica, coloração com vermelho do Congo e microscopia eletrônica de transmissão (TEM).Vesículas e bolhas hemorrágicas estavam presentes na mucosa da cavidade oral, lábios, esôfago e estômago, e, em menor quantidade, no focinho, face e regiões mucocutâneas do períneo, onde a presença de tecido cicatricial era evidente. A avaliação histopatológica e a coloração com vermelho do Congo confirmaram a presença de depósitos de substância amiloide na derme e submucosa, associados à formação de vesículas e bolhas, compatível com amiloidose bolhosa. A TEM confirmou a deposição de fibrila amiloide na derme e ao longo da zona da membrana basal. Observou-se a presença de fissuras na derme superficial e submucosa, justificando a hemorragia e as cicatrizes. A presença de uma gamopatia policlonal e a rápida descoloração do vermelho do Congo após pré-tratamento corroboraram com a amiloide A sérica e a amiloidose secundária. CONCLUSÃO E IMPORTÂNCIA CLÍNICA: A amiloidose bolhosa é uma nova doença descrita em equinos e representa um diagnóstico diferencial recentemente reconhecido para quadros bolhosos, sendo que a natureza hemorrágica das bolhas, presença de cicatrizes e úlceras profundas secundárias são consideradas indícios clínicos da condição.

Ulcerative colitis is a chronic, relapsing and remitting ­inflammatory disease of the colon and rectum. Effective ­management requires prompt recognition and treatment of those with acute relapses as well as appropriate choice and monitoring of drugs for maintenance of remission. This therefore involves specialist gastroenterology teams as well as acute and general physicians and primary care clinicians. Treatment options need to be discussed with those with ­ulcerative colitis and their families and appropriate information provided. This concise guideline focuses (with the generalist particularly in mind) on recommendations from the National Institute for Health and Care Excellence clinical guideline 166 considered of key importance for implementation.

This paper details the development of BEAST Plus, a major upgrade to ABSL's Battery Electrical Analysis Software Tool (BEAST 2007). The upgrade is split into two key areas of focus: characterisation of the NL cell electrical performance across its usable lifetime and implementation of a mathematical solver using a distributed nodal model. In this paper the need for an improved modelling tool is reviewed and the parallel activities of cell characterisation and solver development are discussed, demonstrating the key performance improvements.

Introduction: GM1 gangliosidosis is a rare autosomal recessive genetic disorder caused by the disruption of the GLB1 gene that encodes β-galactosidase, a lysosomal hydrolase that removes β-linked galactose from the non-reducing end of glycans. Deficiency of this catabolic enzyme leads to the lysosomal accumulation of GM1 and its asialo derivative GA1 in β-galactosidase deficient patients and animal models. In addition to GM1 and GA1, there are other glycoconjugates that contain β-linked galactose whose metabolites are substrates for β-galactosidase. For example, a number of N-linked glycan structures that have galactose at their non-reducing end have been shown to accumulate in GM1 gangliosidosis patient tissues and biological fluids. Objective: In this study, we attempt to fully characterize the broad array of GLB1 substrates that require GLB1 for their lysosomal turnover. Results: Using tandem mass spectrometry and glycan reductive isotope labeling with data-dependent mass spectrometry, we have confirmed the accumulation of glycolipids (GM1 and GA1) and N-linked glycans with terminal beta-linked galactose. We have also discovered a novel set of core 1 and 2 O-linked glycan metabolites, many of which are part of structurally-related isobaric series that accumulate in disease. In the brain of GLB1 null mice, the levels of these glycan metabolites increased along with those of both GM1 and GA1 as a function of age. In addition to brain tissue, we found elevated levels of both N-linked and O-linked glycan metabolites in a number of peripheral tissues and in urine. Both brain and urine samples from human GM1 gangliosidosis patients exhibited large increases in steady state levels for the same glycan metabolites, demonstrating their correlation with this disease in humans as well. Conclusions: Our studies illustrate that GLB1 deficiency is not purely a ganglioside accumulation disorder, but instead a broad oligosaccharidosis that include representatives of many β-linked galactose containing glycans and glycoconjugates including glycolipids, N-linked glycans, and various O-linked glycans. Accounting for all β-galactosidase substrates that accumulate when this enzyme is deficient increases our understanding of this severe disorder by identifying metabolites that may drive certain aspects of the disease and may also serve as informative disease biomarkers to fully evaluate the efficacy of future therapies. Keywords: GM1 gangliosidosis, GLB1, Beta-galactosidase, Glycan metabolites, Disease biomarkers, Glycoanalysis

Economic theory is a mathematically rich field in which there are opportunities for the formal analysis of singularities and catastrophes. This article looks at the historical context of singularities through the work of two eminent Frenchmen around the late 1960s and 1970s. Ren\'e Thom (1923-2002) was an acclaimed mathematician having received the Fields Medal in 1958, whereas G\'erard Debreu (1921-2004) would receive the Nobel Prize in economics in 1983. Both were highly influential within their fields and given the fundamental nature of their work, the potential for cross-fertilisation would seem to be quite promising. This was not to be the case: Debreu knew of Thom's work and cited it in the analysis of his own work, but despite this and other applied mathematicians taking catastrophe theory to economics, the theory never achieved a lasting following and relatively few results were published. This article reviews Debreu's analysis of the so called ${\it regular}$ and ${\it crtitical}$ economies in order to draw some insights into the economic perspective of singularities before moving to how singularities arise naturally in the Nash equilibria of game theory. Finally a modern treatment of stochastic game theory is covered through recent work on the quantal response equilibrium. In this view the Nash equilibrium is to the quantal response equilibrium what deterministic catastrophe theory is to stochastic catastrophe theory, with some caveats regarding when this analogy breaks down discussed at the end., Comment: 23 pages, 5 figures

Objective Understanding the pathogenesis of systemic sclerosis (SSc) is confounded by considerable disease heterogeneity. Animal models of SSc that recapitulate distinct subsets of disease at the molecular level have not been delineated. We applied interspecies comparative analysis of genomic data from multiple mouse models of SSc and patients with SSc to determine which animal models best reflect the SSc intrinsic molecular subsets. Methods Gene expression measured in skin from mice with sclerodermatous graft-versus-host disease (GVHD), bleomycin-induced fibrosis, Tsk1/+ or Tsk2/+ mice was mapped to human orthologs and compared to SSc skin biopsy–derived gene expression. Transforming growth factor β (TGFβ) activation was assessed using a responsive signature in mice, and tumor necrosis factor receptor superfamily member 12A (TNFRSF12A) expression was measured in SSc patient and mouse skin. Results Gene expression in skin from mice with sclerodermatous GVHD and bleomycin-induced fibrosis corresponded to that in SSc patients in the inflammatory molecular subset. In contrast, Tsk2/+ mice showed gene expression corresponding to the fibroproliferative SSc subset. Enrichment of a TGFβ-responsive signature was observed in both Tsk2/+ mice and mice with bleomycin-induced skin fibrosis. Expression of TNFRSF12A (the TWEAK receptor/fibroblast growth factor–inducible 14) was elevated in skin from patients with fibroproliferative SSc and the skin of Tsk2/+ mice. Conclusion This study reveals similarities in cutaneous gene expression between distinct mouse models of SSc and specific molecular subsets of the disease. Different pathways underlie the intrinsic subsets including TGFβ, interleukin-13 (IL-13), and IL-4. We identify a novel target, Tnfrsf12a, with elevated expression in skin from patients with fibroproliferative SSc and Tsk2/+ mice. These findings will inform mechanistic and translational preclinical studies in SSc.

Introduction Inflammatory Bowel Disease (IBD) services across the UK are under increasing pressure. To improve efficiency, pathways of care have been proposed. However these are difficult to cost due to a lack of data on current service provision, leading to challenges with commissioning. We set out to characterise our IBD service in a District General Hospital (DGH) setting, having recently implemented a new pathway to streamline the service. A second aim was to establish if our service met the NICE quality standard of seeing IBD referrals within 4 weeks. Methods Prospective data from clinics at two DGHs were gathered from 92 patient journeys over 52 weeks. The activity of a new IBD helpline was analysed over a 28 week period and outcomes prospectively recorded. Specifically the activity avoided as a result of the helpline was analysed and costed. Finally, the outcomes from a new virtual IBD clinic were prospectively collected over a 10 week period. Results 33% of clinic patients had IBD, of which 59% were in remission. 41% of patients were felt suitable for non-clinic follow-up. 76% were interested in the concept of ‘self-management’ during remission. 95% of patients rated the consultation experience as ‘good’ or ‘very good’. There was an average 1 new to 4 follow-up encounters within the first year from referral; 26% were successfully conducted by telephone. Median time from initial referral to first outpatient contact was 9.1 weeks (4.9–19.9). 58% were not seen within 4 weeks of referral. The IBD helpline received 543 calls in 28 weeks (average 83/month). The interventions avoided due to this service were; 156 GP consultations, 231 outpatient reviews, 39 ED attendances and 6 admissions. Estimated cost saving due to activity avoided was £37,913, with helpline costs of £2065. 92% of patients rated the helpline as ‘good’ or ‘very good’. 100% of patients reported a response within 24 hours (weekdays). Since September 2017, 10 virtual IBD clinics have led to 191 treatment decisions. High cost biologic drugs have been stopped in 7 patients, without having to wait for face-to-face review, with an estimated saving of £12 300. 59 patients were identified as needing annual review and 51 identified as needing urgent treatment decisions. Overall quantifiable cost savings from the helpline and virtual clinic totalled £11,000/month. Conclusions Telephone and virtual clinics result in earlier treatment decisions for IBD patients and give quantifiable cost savings as part of an IBD pathway at a DGH. By increasing virtual clinics and encouraging patient autonomy, we can aim to improve the NICE quality standard of seeing new IBD patients within 4 weeks of referral. Improving autonomy over services for both clinician and patient can allow for more efficient IBD care.

We use methods of complex analysis to extend the bundle structure across a removable point-singularity in a Sasakian three-manifold., Comment: 14 pages