Extracellular vesicle proteomes of two transmissible cancers of Tasmanian devils reveal tenascin-C as a serum-based differential diagnostic biomarker.

The iconic Tasmanian devil (Sarcophilus harrisii) is endangered due to the transmissible cancer Devil Facial Tumour Disease (DFTD), of which there are two genetically independent subtypes (DFT1 and DFT2). While DFT1 and DFT2 can be differentially diagnosed using tumour biopsies, there is an urgent need to develop less-invasive biomarkers that can detect DFTD and distinguish between subtypes. Extracellular vesicles (EVs), the nano-sized membrane-enclosed vesicles present in most biofluids, represent a valuable resource for biomarker discovery. Here, we characterized the proteome of EVs from cultured DFTD cells using data-independent acquisition-mass spectrometry and an in-house spectral library of > 1500 proteins. EVs from both DFT1 and DFT2 cell lines expressed higher levels of proteins associated with focal adhesion functions. Furthermore, hallmark proteins of epithelial-mesenchymal transition were enriched in DFT2 EVs relative to DFT1 EVs. These findings were validated in EVs derived from serum samples, revealing that the mesenchymal marker tenascin-C was also enriched in EVs derived from the serum of devils infected with DFT2 relative to those infected with DFT1 and healthy controls. This first EV-based investigation of DFTD increases our understanding of the cancers' EVs and their possible involvement in DFTD progression, such as metastasis. Finally, we demonstrated the potential of EVs to differentiate between DFT1 and DFT2, highlighting their potential use as less-invasive liquid biopsies for the Tasmanian devil.

Death Associated With Nadolol for Infantile Hemangioma: A Case for Improving Safety.

Nadolol is a ß-adrenergic antagonist that has been shown to be efficacious in the treatment of infantile hemangioma. It has been suggested that this drug may have fewer side effects compared with the gold standard therapy, propranolol, because it does not exhibit membrane-stabilizing effects and has little ability to cross the blood-brain barrier. However, the pharmacokinetics and safety of nadolol in infants are not well understood, potentially making this therapy dangerous. ß-adrenergic antagonist toxicity causes bradycardia, hypotension, hypoglycemia, and even death. We report a case of a 10-week-old girl who was started on nadolol for infantile hemangioma, died 7 weeks later, and was found to have an elevated postmortem cardiac blood nadolol level of 0.94 mg/L. The infant had no bowel movements for 10 days before her death, which we hypothesize contributed to nadolol toxicity. Pharmacokinetics studies show a large fraction of oral nadolol either remains in the feces unchanged or is excreted into feces via the biliary system, allowing continued absorption over time in infants who stool infrequently. Propranolol may be a safer therapy overall. Not only does it have a shorter half-life, but propranolol is hepatically metabolized and renally eliminated, allowing for less drug accumulation in healthy infants with variable stooling patterns. We suggest that if nadolol is selected for therapy, pediatricians should instruct parents to monitor their infants' bowel movements closely and encourage early intervention in the event of decreased stooling. This intervention may greatly improve the safety of nadolol in this vulnerable patient population.

Clinical Usefulness of the Platelet-to Lymphocyte Ratio in Patients with Angiosarcoma of the Face and Scalp.

Angiosarcoma of the face and scalp (ASFS) is an extremely aggressive tumor that frequently metastasizes, often leading to death. The neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR) are inflammatory markers that predict outcome of various cancers. We aimed to examine the relationship between pretreatment inflammatory markers and ASFS outcome. We included 17 patients with ASFS and a control group of 56 age- and gender-matched healthy individuals. Total white blood counts, neutrophil, lymphocyte, monocyte, and platelet counts were recorded; NLR, PLR, and LMR were calculated. Kaplan-Meier curves were used to calculate overall survival (OS) and distant metastasis-free survival (DMFS). Optimal cut-off values for each inflammatory marker were calculated using receiver operating curve analysis. Median follow-up was 22 months (range, 6-75). There was a statistically significant difference in absolute neutrophil counts and NLR between patient and control groups. Two-year OS and DMFS rates were 41% and 35%, respectively. In patients with tumors < 10 cm, PLR was highly correlated with DMFS, with the 2-year DMFS for those with a high PLR being 50% compared with 100% for those with a low PLR (p = 0.06). This study suggests that PLR is superior to NLR and LMR, and is a clinically useful marker in patients with ASFS with small tumors.

ERBB3: A potential serum biomarker for early detection and therapeutic target for devil facial tumour 1 (DFT1).

Devil Facial Tumour 1 (DFT1) is one of two transmissible neoplasms of Tasmanian devils (Sarcophilus harrisii) predominantly affecting their facial regions. DFT1's cellular origin is that of Schwann cell lineage where lesions are evident macroscopically late in the disease. Conversely, the pre-clinical timeframe from cellular transmission to appearance of DFT1 remains uncertain demonstrating the importance of an effective pre-clinical biomarker. We show that ERBB3, a marker expressed normally by the developing neural crest and Schwann cells, is immunohistohemically expressed by DFT1, therefore the potential of ERBB3 as a biomarker was explored. Under the hypothesis that serum ERBB3 levels may increase as DFT1 invades local and distant tissues our pilot study determined serum ERBB3 levels in normal Tasmanian devils and Tasmanian devils with DFT1. Compared to the baseline serum ERBB3 levels in unaffected Tasmanian devils, Tasmanian devils with DFT1 showed significant elevation of serum ERBB3 levels. Interestingly Tasmanian devils with cutaneous lymphoma (CL) also showed elevation of serum ERBB3 levels when compared to the baseline serum levels of Tasmanian devils without DFT1. Thus, elevated serum ERBB3 levels in otherwise healthy looking devils could predict possible DFT1 or CL in captive or wild devil populations and would have implications on the management, welfare and survival of Tasmanian devils. ERBB3 is also a therapeutic target and therefore the potential exists to consider modes of administration that may eradicate DFT1 from the wild.

Discovery of Biomarkers for Tasmanian Devil Cancer (DFTD) by Metabolic Profiling of Serum.

Devil facial tumor disease (DFTD) is a transmissible cancer threatening Tasmanian devils (Sarcophilus harrisii) with extinction. There is no preclinical test available for DFTD, and thus our aim was to find biomarkers for DFTD by metabolic fingerprinting. Seventy serum samples from wild Tasmanian devils (35 controls, 35 with tumors) were analyzed by liquid chromatography-high-resolution mass spectrometry. Features were selected by multivariate models (PLS/DA, random forests) comparing age-matched training set (n = 20 × 2) and further complying with fold-change threshold (≥1.4) and Mann-Whitney U-tests with correction for multiple hypotheses (false discovery rate (FDR) q < 0.05). An array of overlapping peptide segments of the N-terminal end of fibrinogen were the strongest positive DFTD markers. These peptides recorded fold-change up to 90, FDR-corrected p value below 0.01, and area under ROC curve of at least 0.80 and also correlated with tumor size (Spearman R > 0.45, p < 0.01). Additional potential markers included amino acid and lipid metabolites, while cortisol and urea were the most significant health predictors (AUC ≥ 0.90). PLS/DA resulted in AUC = 0.997 for the training set and overall sensitivity of 91% and specificity of 97%. A support vector machine model utilizing only the major peptide marker and seven other metabolites led to overall 94% sensitivity and specificity. The novel findings in this first DFTD metabolomics study shed light on metabolic changes in Tasmanian devils affected by DFTD and provide a valuable step toward the development of prognostic biomarkers.

Hematologic and serum biochemical changes associated with Devil Facial Tumor Disease in Tasmanian Devils.

BACKGROUND: Devil Facial Tumor Disease (DFTD) is an infectious tumor causing significant population declines in wild Tasmanian Devils. While clinical assessment and pathology have been well reported for DFTD, there is a lack of information on hematologic and biochemical alterations associated with DFTD. OBJECTIVES: The purpose of the study was to determine hematologic and serum biochemical variation in healthy, wounded, and DFTD-affected Tasmanian Devils. METHODS: Blood samples were collected from wild Tasmanian Devils at 5 sites in Tasmania. Hematology and clinical biochemistry variables were compared between clinically healthy, wounded, and DFTD-affected devils. Differences were also analyzed among stages of DFTD, including individuals pre- and postclinical signs developing, and between ulcerated and nonulcerated DFTD tumors. RESULTS: Statistically significantly increased counts in WBC, neutrophils, and platelets, and concentration of fibrinogen, as well as decreased counts in lymphocytes, erythrocytes, and HGB concentration were observed in DFTD-affected devils compared to healthy devils. Activities of ALP, ALT, and GLDH, concentrations of sodium, potassium and albumin, and sodium-to-potassium ratio and albumin-to-globulin ratio were significantly lower, and AST activity was significantly higher in animals with DFTD when compared to clinically healthy animals. No significant differences were found among stages of DFTD or ulcerated and nonulcerated tumors. CONCLUSIONS: The differences in hematology and clinical chemistry variables in devils with DFTD compared to healthy devils are nonspecific and reflective of acute phase response and inflammation, and anemia of chronic disease. Similar changes are observed with wounds but to a lesser extent.

Analysis of serum and salivary immunoglobulin M in patients with orofacial epithelial cancers.

CONTEXT: The physiological changes in the humoral immune system of patients with orofacial epithelial cancers (OECs) are considered key factors in the pathogenesis, prognosis, and management of these individuals. AIM: This study assessed the serum and salivary immunoglobulin M (IgM) levels in patients with OECs. SETTINGS AND DESIGNS: This is a cross-sectional study of the serum and salivary IgM profile among patients with OEC and healthy controls. MATERIALS AND METHODS: There were 78 subjects comprising 30 patients with untreated OEC, 18 patients with OEC receiving treatment and 30 healthy, age and gender matched individuals. The serum and salivary samples from the participants were analyzed for IgM using the enzyme linked immunosorbent assay technique. RESULTS: The mean value of serum IgM in OEC patients receiving treatment was significantly lower compared to untreated OEC patients and healthy controls (P = 0.01). However, the mean serum IgM among untreated OEC patients was not significantly different compared with healthy controls. In contrast, the salivary IgM level did not show any significant difference among the three groups (P = 0.06). Furthermore, there was no correlation between the serum and salivary levels of IgM among the subjects. CONCLUSION: The findings from this study suggest that serum IgM levels in OEC patients receiving treatment might be good biomarker while salivary IgM may not be reliable as a marker in these individuals.

The effects of season and devil facial tumour disease on the reproductive physiology of the male Tasmanian devil (Sarcophilus harrisii).

Devil facial tumour disease (DFTD) is the cause of the rapid decline of wild Tasmanian devils. Female devils are seasonal breeders with births peaking during autumn (i.e. March) but the degree of reproductive seasonality in male devils is unknown. The objective of this study was to examine the potential effects of season and DFTD on reproductive function in male devils (n=55). Testicular (1.90±0.23 g) and epididymal (0.90±0.06 g) weights were maximal during autumn and spring (P<0.05), whereas prostate (3.71±0.74 g) and Cowper's gland (0.68±0.22; 0.52±0.21 g) weights peaked during autumn (P<0.001). The motility of spermatozoa from the cauda epididymides extracted post-mortem was similar (P>0.05) across season and disease state (31.5±13.1% total motility). Testicular and epididymal weights were no different between animals displaying late or early-stage DTFD signs or disease-free animals (P>0.1). The accessory sex glands were larger in late-stage DFTD animals than in animals with early-stage disease signs or which were disease-free (P<0.01) but effects of season on this result can't be excluded. Serum testosterone concentrations peaked during summer (0.25±0.18 ng mL(-1)) but values were not different from the preceding and subsequent seasons (P>0.05), nor influenced by disease stage (P>0.1). Seasonal and DFTD-related changes in serum cortisol concentrations were not evident (P>0.1). Male devil reproduction does not appear to be restricted by season nor inhibited by DFTD.

Estimation of vascular endothelial growth factor and placental growth factor serum levels' in infant with hemangioma and population of healthy infants.

INTRODUCTION: Hemangioma is found in approximately 10% of infants as the most prevalent benign neoplasm. The natural history of hemangioma is typical for this lesion and includes two phases: fast growth during the first year of life of the child and subsequent slow regression lasting some five years. Even though the etiopathogenesis of hemangioma has not been fully elucidated, the role played in this process by vascular growth factors remains unquestionable. The aim of this work was to assess the value of serum levels of the vascular endothelial growth factor (VEGF) and placental-derived growth factor (PlGF) for therapy planning in infants with hemangiomas. MATERIAL AND METHODS: The study group comprised 43 infants, aged 2 weeks to 6 months, with hemangiomas on the body. 25 girls and 11 boys participated in the second stage of the study done 14 months later. We analyzed correlations between serum levels of vascular growth factors and phase of hemangioma, clinical symptoms, and findings in ultrasonography with Power Doppler visualization. Normal ranges for VEGF and PlUF were established for healthy infants. RESULTS: The results in the study group were analyzed statistically and presented as arithmetic means, standard deviations, medians, minimal and maximal values, and percentage distributions. CONCLUSIONS: In local population of healthy infants the ranges of VEGF and P1GF serum levels are very wide; VEGF and P1GF serum levels determined in infants affected with hemangioma do not reflect the dynamics of observable lesion's evolution.

Clearance of ascorbic acid in plasma in patients before major maxillofacial surgery compared with that in volunteers.

Reduced concentration of ascorbic acid (AA) in plasma is often observed in tumor patients. Besides malnutrition, an increased AA consumption in tumor patients is discussed as a main reason for reduced AA concentration in plasma. A reduced AA concentration caused by malnutrition would not influence the total clearance (Cltot) of AA in plasma; however, a change in AA consumption would alter the Cltot. To investigate this further, a study was carried out on 20 patients hospitalized for major maxillofacial surgery and 20 volunteers. The Cltot of AA in plasma after bolus injection of AA (4 mg/kg body wt) in tumor patients was compared with that in volunteers. AA concentration in plasma (P < 0.05) was significantly lower in tumor patients (median and 25th and 75th percentiles: 23.7, 14.6, and 32.7 mumol/l) than in volunteers (median and 25th and 75th percentiles: 66.5, 59.6, and 78.6 mumol/l). No significant change in Cltot was observed for tumor patients (median and 25th and 75th percentiles: 11.5, 8.0, and 14.0 l/h) compared with volunteers (median and 25th and 75th percentiles: 11.3, 9.9, and 13.2 l/h). No indication of an increased consumption of AA in tumor patients could be detected. The lowered concentration of AA in plasma observed in our patients might be the result of a reduced AA intake.

Cutaneous angiosarcoma with thrombocytopenia.

Thrombocytopenia was observed in 3 patients with cutaneous angiosarcoma of the scalp and face. A sudden decrease in the platelet count occurred in association with rapid enlargement of primary or metastatic lesions. Neither antiplatelet antibody nor platelet-associated IgG was detected. Increased serum levels of beta-thromboglobulin and platelet factor 4 indicated that platelets were destroyed and consumed within the vascular bed of the tumor. Prominent PECAM-1 expression on tumor cells may be involved in intratumoral platelet aggregation and consumption. We suggest that the sudden development of profund thrombocytopenia in patients with angiosarcoma may suggest either rapid growth of the primary tumor or herald the development of metastatic disease.

[Burkitt lymphoma in Cameroon children: descriptive and clinico-anatomical aspects]. / Lymphome de Burkitt chez l'enfant au Cameroun: aspects descriptifs et anatomocliniques.

This retrospective study is armed to indicate the descriptive and pathological aspects of children Burkitt lymphoma in Cameroon. It was performed on a 4 year period between July 1988 and July 1992. Children from 0 to 15-year-old who were hospitalized and who had histologically provern Burkitt lymphoma were included. There were 39 patients, that is 27% of all malignant tumors in children during this period. Twenty-four were boys and 15 were girls; the median age was 90 months (+/-46) (range from 3 to 180 months). All children had Plasmodium falciparum infection. EBV serology was positive in 18 patients out of 25 (72%), 14 (36%) had a good nutritional status, the 25 other patients suffered malnutrition. Tumor localizations were: maxillary in 29 (74%) patients, abdominal in 7 (18%), other in 3 patients. Clinical stages according to Murphy classification were: stage I in 6 (15%) patients, II in 3 (8%), III in 20 (51%) and IV in 10 (26%). It is concluded from this series that clinical aspects and histological pattern in children Burkitt lymphoma in Cameroon are not different from what is observed in other endemic areas.

Applicability of the general nutritional status score to patients with oral and maxillofacial malignancies.

The general nutritional status (GNS) score was used to assess the nutritional status of 127 consecutive patients with oral and maxillofacial malignancies. Forty-six of our patients (36.2%) were undernourished (group 2), while 81 (63.8%) were in good nutritional condition (group 1). Comparison of nutritional laboratory tests between these two groups showed that the differences in serum albumin, transferrin, and creatinine-height index (CHI) had statistical significance, yielding an excellent correlation between the GNS score and the nutritional laboratory test. Undernourished patients had a significantly higher postoperative complication incidence (48.3%) than well-nourished patients (19.4%) (chi 2 = 6.637; P < 0.01), indicating that the GNS score can be used as a prognostic index.

[Hemostatic changes in patients with tumors of the maxillofacial area]. / Izmeneniia gemostaza u bol'nykh s opukholiami cheliustno-litsevoi oblasti.

To study hemostatic impairments in patients with maxillofacial tumors, 131 patients and 30 healthy subjects were examined. The patients were found to develop early hemostatic changes which appeared as a latent DIC syndrome. The mobility of the physiological system of PASA permits the equilibrium of coagulative and anticoagulative mechanisms to be preserved to a definite extent and fairly long. The changes were detected during the formation of prothrombinase, thrombin and during fibrin formation and stabilization in the fibrinolytic system. Hypercoagulation typical of the DIC syndrome at the early stages of tumor was followed by consumption coagulopathic signs and profuse bleeding risk at the T4 and N3 stages.

Distribution of adhesion molecules, IgE, and CD23 in a case of angiolymphoid hyperplasia with eosinophilia.

Angiolymphoid hyperplasia with eosinophilia developed in a 13-year-old boy. The immune phenotype of infiltrating cells and the distribution of adhesion molecules of the integrin, immunoglobulin, and selectin families were analyzed with monoclonal antibodies. The infiltrate consisted mainly of helper lymphocytes, monocytes, eosinophils, CD1+ dendritic cells, and mast cells. A CD23/Fc epsilon receptor was present on 40% of these cells, and surface bound IgE was present on 30% of these cells. The latter was found on all cell types. A cellular adhesion molecule analysis revealed that the proliferating endothelial cells were expressing constitutive ICAM-1, LFA-3, VLA-1, VLA-3, VLA-5, alpha 6, and beta 3-integrin chain as well as activation-dependent VLA-2, VCAM-1, and human lymphocyte antigen-DR. The infiltrating cells expressed LFA-1 and CD2, the correlating ligands for ICAM-1 and LFA-3. Our studies support previous observations of an unusual IgE-mediated immune reaction, and suggest an additional pathogenic role for adhesion events in the development of the pathognomonic cellular composition in angiolymphoid hyperplasia with eosinophilia.

Serum sialic acid levels in patients with oral and maxillofacial malignancy.

Serum sialic acid levels were measured using the thiobarbituric acid method in 80 healthy subjects and in 40 patients with benign and 61 with malignant tumors in the oral and maxillofacial region. It was shown that the cancer group had statistically higher sialic acid levels than control group and the group with benign disease (P less than .05), but no significant difference was noted between the normal control group and the patients with benign tumor (P greater than .05). Serum sialic acid levels were significantly higher in the patients with stages III and IV cancer than in those individuals with stage I and II cancer, but no differences were seen between stages I and II, and between stages III and IV cancer patients (P greater than .05). There was also no difference when considering the anatomic site of the malignant lesion. Furthermore, during the serial sialic acid determination in cancer patients before and following treatment, the sialic acid levels usually declined with remission of the disease and increased with recurrence, metastasis, or a poor prognosis. We conclude from this study that the sequential measurement of sialic acid level is particularly useful in monitoring patients with oral cancer, and sialic acid may prove to be a valuable tumor marker in oral and maxillofacial malignancy.