Citrullination Was Introduced into Animals by Horizontal Gene Transfer from Cyanobacteria.

Protein posttranslational modifications add great sophistication to biological systems. Citrullination, a key regulatory mechanism in human physiology and pathophysiology, is enigmatic from an evolutionary perspective. Although the citrullinating enzymes peptidylarginine deiminases (PADIs) are ubiquitous across vertebrates, they are absent from yeast, worms, and flies. Based on this distribution PADIs were proposed to have been horizontally transferred, but this has been contested. Here, we map the evolutionary trajectory of PADIs into the animal lineage. We present strong phylogenetic support for a clade encompassing animal and cyanobacterial PADIs that excludes fungal and other bacterial homologs. The animal and cyanobacterial PADI proteins share functionally relevant primary and tertiary synapomorphic sequences that are distinct from a second PADI type present in fungi and actinobacteria. Molecular clock calculations and sequence divergence analyses using the fossil record estimate the last common ancestor of the cyanobacterial and animal PADIs to be less than 1 billion years old. Additionally, under an assumption of vertical descent, PADI sequence change during this evolutionary time frame is anachronistically low, even when compared with products of likely endosymbiont gene transfer, mitochondrial proteins, and some of the most highly conserved sequences in life. The consilience of evidence indicates that PADIs were introduced from cyanobacteria into animals by horizontal gene transfer (HGT). The ancestral cyanobacterial PADI is enzymatically active and can citrullinate eukaryotic proteins, suggesting that the PADI HGT event introduced a new catalytic capability into the regulatory repertoire of animals. This study reveals the unusual evolution of a pleiotropic protein modification.

Eosinophilic globules in a classic ependymoma: evidence of a possible secretory role.

A number of neoplasms of the central nervous system can demonstrate diffuse eosinophilic globules, known to be secretory products of the corresponding cell type, but they have not been a salient feature in descriptions of classic ependymoma. Here, we present a case of a posterior fossa ependymoma demonstrating glassy PAS-positive, diastase-resistant, eosinophilic globules with light microscopic and ultrastructural features resembling Reissner fiber, the secretory product of the subcommissural organ. While there has been a single published description of an ependymoma with intra- and extracellular granulofibrillary material suggested to be evidence of secretory differentiation, ours is the first case to demonstrate diffuse eosinophilic globules in an ependymoma. The extent of globules allowed full study by electron microscopy to provide new insight into the secretory material and the surrounding structures. Our findings suggest that neoplastic ependymal cells can recapitulate the secretory capacity of the subcommissural organ.

Nerve growth factor inhibition with tanezumab influences weight-bearing and subsequent cartilage damage in the rat medial meniscal tear model.

OBJECTIVE: To investigate whether the effects of nerve growth factor (NGF) inhibition with tanezumab on rats with medial meniscal tear (MMT) effectively model rapidly progressive osteoarthritis (RPOA) observed in clinical trials. METHODS: Male Lewis rats underwent MMT surgery and were treated weekly with tanezumab (0.1, 1 or 10 mg/kg), isotype control or vehicle for 7, 14 or 28 days. Gait deficiency was measured to assess weight-bearing on the operated limb. Joint damage was assessed via histopathology. A second arm, delayed onset of treatment (starting 3-8 weeks after MMT surgery) was used to control for analgesia early in the disease process. A third arm, mid-tibial amputation, evaluated the dependency of the model on weight-bearing. RESULTS: Gait deficiency in untreated rats was present 3-7 days after MMT surgery, with a return to normal weight-bearing by days 14-28. Prophylactic treatment with tanezumab prevented gait deficiency and resulted in more severe cartilage damage. When onset of treatment with tanezumab was delayed to 3-8 weeks after MMT surgery, there was no increase in cartilage damage. Mid-tibial amputation completely prevented cartilage damage in untreated MMT rats. CONCLUSIONS: These data suggest that analgesia due to NGF inhibition during the acute injury phase is responsible for increased voluntary weight-bearing and subsequent cartilage damage in the rat MMT model. This model failed to replicate the hypotrophic bone response observed in tanezumab-treated patients with RPOA.

Endodermal Cyst of the Third Nerve in a Child.

Endodermal cysts, also known as neurogenic, neuroenteric, foregut, bronchogenic, respiratory, epithelial, teratomatous, or gastrocytoma cysts, can be found in the central nervous system, predominantly in the subarachnoid space of the cervical and thoracic spinal cord. We describe a child with an endodermal cyst of the third nerve and highlight neuroimaging findings, pathological correlation, clinical course, and patient management.

Pathogenicity and Aggressiveness of Three Alternaria spp. on Potato Foliage in the U.S. Northwest.

Alternaria spp. were collected from potato foliage showing symptoms of early blight and brown spot in the Columbia Basin, WA and Bonners Ferry and Rupert, ID between 2009 and 2011. The aggressiveness of three Alternaria spp. on potato was quantified on nonwounded and wounded detached leaves of 'Russet Norkotah' potato; wounded detached leaves of 'Alturas', 'Ranger Russet', 'Russet Burbank', and 'Umatilla Russet'; and whole plants of Russet Norkotah. Mean infection frequencies (MIF) and area under the lesion expansion curve (AULEC) were significantly greater for Alternaria solani (P = 0.0072 and 0.0002, respectively) than for A. arborescens or A. arbusti on nonwounded leaves. Wounding of tissue significantly increased MIF and AULEC for A. arbusti (P = 0.008 and 0.0047, respectively) and AULEC for A. arborescens (P = 0.01) relative to nonwounded tissue. AULEC did not differ significantly among the three Alternaria spp. when inoculated onto wounded foliage of whole plants (P = 0.34); the AULEC of whole plants was positively and significantly correlated with AULEC on detached leaves (P = 0.03). Umatilla Russet was the most susceptible and Russet Burbank was the least susceptible based on MIF and AULEC for all three pathogen species. Results indicate that A. solani was the more aggressive pathogen of potato in the Columbia Basin, because both A. arborescens and A. arbusti require wounds and A. arbusti lesions do not expand significantly in comparison with A. solani or A. arborescens following inoculation.

Rhabdomyomatous mesenchymal hamartoma presenting as a sacral skin tag in two neonates with spinal dysraphism.

Rhabdomyomatous mesenchymal hamartoma (RMH) is a rare congenital malformation involving the dermis and subcutaneous tissue, of which there were 62 reported cases through 2014. We report RMH in two neonates presenting as a sacral skin tag. In both cases, magnetic resonance imaging (MRI) of the spine showed evidence of spinal dysraphism, including a lipomyelomeningocele and a tethered cord. Surgical repair of the defects was performed. Histopathologic examination of the skin tags showed a haphazard arrangement of mature skeletal muscle fibers and adnexal elements, consistent with RMH. The second patient also had a hemangioma on the sacrum and was diagnosed with LUMBAR (lower body hemangioma and other cutaneous defects, urogenital anomalies/ulceration, myelopathy, bony deformities, anorectal/arterial anomalies, and renal anomalies) syndrome, an association between cutaneous infantile hemangiomas of the lower body and regional congenital anomalies. The apparent association of paraspinal RMH with spinal dysraphism suggests that aberrant migration of mesodermally derived tissues (including skeletal muscle fibers) during neural tube development may be responsible for the pathologic findings in the skin. Additional study of patients with spinal dysraphism and congenital cutaneous lesions may further support this hypothesis.

An Extraordinary Association of Glomus Tumor and Pacinian Hyperplasia in the Hand of a Female Patient.

BACKGROUND: Glomus tumor is a benign neoplasm of the glomus body, a neuromyoarterial structure that regulates temperature and pressure in the cutaneous vasculature. Approximately 1%-4.5% of glomus tumors present in the hands of females; of these, 65% are seen in the subungual region of the index and long fingers. Pacinian hyperplasia is a benign lesion of the Pacinian corpuscle, a mechanoreceptor located in the subcutis of the hands and feet. METHODS: A 65-year-old woman with a history of hand trauma and a 1-year chief complaint of tingling, pain, and burning sensations in her proximal thumb underwent exploration of the digital nerve after an x-ray and 2 magnetic resonance imaging examinations failed to detect a mass. Two lesions immediately adjacent to each other were excised. RESULTS: Microscopic examination showed Pacinian hyperplasia, and a second proliferation of solid epithelioid cells related to benign blood vessels. Immunohistochemistry confirmed the epithelioid cells to be strongly positive for smooth muscle actin, CD34, and type 4 collagen, which is consistent with the phenotype of a glomus tumor. The cells were negative for S100 protein. CONCLUSIONS: The association of glomus tumor with Pacinian hyperplasia has rarely been reported in the literature. We present another rare case to bring awareness to this differential diagnostic consideration.

Diagnostic Algorithm for Patients With Suspected Giant Cell Arteritis.

BACKGROUND: To identify clinical and laboratory factors contributing to the diagnosis of giant cell arteritis (GCA) and develop a diagnostic algorithm for the evaluation of GCA. METHODS: Retrospective review of 213 consecutive cases of temporal artery biopsy (TAB) seen at a single academic center over a 10-year period (2000-2009). Pathologic specimens were re-reviewed and agreement between the original and second readings was assessed. A composite clinical suspicion score was created by adding 1 point for each of the following criteria: anterior extracranial circulation ischemia, new onset headache, abnormal laboratory results (erythrocyte sedimentation rate, C-reactive protein (CRP), or platelet count), jaw claudication, abnormal or tender superficial temporal artery, constitutional symptoms, and polymyalgia rheumatica; one point was subtracted if a comorbid condition could explain a criterion. RESULTS: Of the 204 TABs analyzed, pathologic findings were confirmatory in 49 (24.0%) and suggestive in 12 (5.9%). TAB-positive patients were more likely to be older (age 75.2 ± 7.8 vs 69.7 ± 11.0 years, P = 0.0002), complain of jaw claudication (relative-risk = 3.26, P = 0.0014), and have thrombocytosis (relative-risk = 3.3, P = 0.0072) and elevated CRP (relative-risk = 1.8, P = 0.037). None of the patients with a clinical score less than 2 had a positive TAB. Diabetes mellitus and kidney disease were often the explanation for the symptoms and abnormal clinical finding(s) that led to a negative TAB. CONCLUSIONS: We propose a clinical algorithm that is highly predictive for a positive TAB and can be valuable in the evaluation process of suspected cases of GCA.

Effect of Powdery Scab Root Galls on Yield of Potato.

Powdery scab of potato, caused by Spongospora subterranea f. sp. subterranea, has increased in incidence since 1981 to become a major concern for potato production in the Columbia Basin of south-central Washington and north-central Oregon. Disease symptoms consist of pustules on the tuber surface and wart-like galls on roots, stolons, and tubers. In the current study, the effect of powdery scab root galls on potato yield was quantified by comparing yields of cultivars susceptible and resistant to root galling grown on soil naturally infested and noninfested with the powdery scab pathogen. Potato tuber yields for the three yield components of yield per plant, number of tubers, and weight of tubers were not negatively affected by powdery scab galls on roots of Umatilla Russet and Shepody, as indicated by yield ratios ≥1.0 in 3 of 4 years, and also by regression analyses in all 3 years. Regression lines with a nonsignificant slope or slopes that significantly increased with gall index also indicated that root galls did not negatively affect yield. Mean tuber weight increased as gall index increased for Umatilla Russet in 2 of 3 years. Consequently, control tactics directed solely at reducing powdery scab galls on roots are not justified economically under conditions encountered in the Columbia Basin of Washington and Oregon states. Effort has been made to develop resistant cultivars and search for soil applied chemicals to reduce root galls.

Effect of Extended Crop Rotations on Incidence of Black Dot, Silver Scurf, and Verticillium Wilt of Potato.

Potato tubers were collected and evaluated for symptoms and signs of black dot, silver scurf, and Verticillium wilt to determine the effect of extended crop rotations on disease incidences in the Columbia Basin. Incidence of tubers with black dot collected from storage significantly decreased as the number of years between potato crops increased from 3 to 5 years and beyond and significantly increased as the number of previous potato crops increased to 16. The highest incidence of black dot (range of 73 to 98%) was from fields rotated out of potatoes for 1 to 3 years. The mean incidence of black dot was 56% for fields out of potatoes for 0 to 4 years and 12% for fields out of potatoes 5 and more years. A low incidence (0 to 9%) of black dot was detected at 15 years out of potatoes. Years out of potato and number of prior potato crops accounted for 71% of the variability associated with the incidence of black dot. Severity of black dot on tuber periderm peels significantly increased as incidence of tuber periderm peels with Colletotrichum coccodes increased. Coefficient of determination was 0.87 for log severity on regressed on black dot incidence. Incidence of silver scurf was highest from fields out of potatoes for 1 year. Incidence of silver scurf infected tubers significantly increased as the number of previous potato crops increased due to short rotations between potato crops. Incidence of tubers with Verticillium dahliae was not related to years between potato crops or number of previous potato crops. The present study confirmed that black dot can be reduced with rotations out of potatoes greater than 5 years.

Accuracy of Rain Forecasts for Use in Scheduling Late Blight Management Tactics in the Columbia Basin of Washington and Oregon.

Accuracy of prediction was analyzed for 17- and 30-day rain forecasts at two locations in the Columbia Basin to determine whether forecasts were sufficiently accurate to be included as a model component to schedule fungicide applications for potato late blight. Accuracy was partitioned into specificity (percentage of forecasted nonrainfall events classified correctly) and sensitivity (percentage of forecasted daily rainfall events classified correctly). An adjusted sensitivity, which included the forecasted rain day plus the next 2 days, was also used to give a wider target than only 1 day for evaluating accuracy of forecasted rain events. For 17-day forecasts, specificity during the seasonal test period was ≥70% from mid-June through September and specificity over the days of the forecast was >70% for the first 8 days at both locations both years. Adjusted sensitivity over days of the forecast was initially >80% and then decreased as forecasts increased from 7 to 17 days for 17-day forecasts at both locations and years. Sensitivity and adjusted sensitivity during the seasonal test period were both positively correlated with the number of rainy days while specificity was negatively correlated. Adjusted sensitivity was considerably higher for May (month with highest incidence of rain) than July (month with lowest incidence of rain) at both locations. For 30-day forecasts, specificity during the test period was >75% in July and August and adjusted sensitivity ranged from 60 to 100% for time periods occurring in May and June during both sample seasons. Specificity was generally above 80% as days of the forecast increased and adjusted sensitivity varied greatly over days of the forecasts, with extremes between 0 and 100% at both locations and years for the 30-day forecasts. Specificity of 17- and 30-day rain forecasts and adjusted sensitivity of 17-day rain forecasts have utility in scheduling late blight fungicides in the Columbia Basin.

Comparative nonclinical assessments of the proposed biosimilar PF-05280586 and rituximab (MabThera®).

Comparative nonclinical studies were conducted with the proposed biosimilar PF-05280586 and rituximab-EU (MabThera®). In side-by-side analyses, peptide maps and complement-dependent cytotoxicity assay results were similar. Sexually-mature cynomolgus monkeys were administered PF-05280586 or rituximab-EU as a single dose of 0, 2, 10, or 20 mg/kg on day 1 and observed for 92 days (single-dose study) or as 5 weekly injections of 0 or 20 mg/kg and necropsied on day 30, the day after the 5th dose, or on day 121 (repeat-dose study). The pharmacokinetic and pharmacodynamic profiles for both molecules were similar. Marked depletion of peripheral blood B cells 4 days after dosing was followed by near or complete repletion (single-dose study) or partial repletion (repeat-dose study). In the single-dose study, anti-drug antibodies (ADA) were detected by day 29 in all animals administered PF-05280586 or rituximab-EU and persisted through day 85, the last day tested. In the repeat-dose study, ADA were detected on day 121 in 50% of animals administered PF-05280586 or rituximab-EU. Both molecules were well tolerated at all doses. In all endpoints evaluated, PF-05280586 exhibited similarity to rituximab-EU.

Melanocytic Nevus of the Superior Conjunctival Fornix: A Case Report.

Introduction: Conjunctival nevi are benign tumors that are commonly located at the nasal or temporal limbus and rarely in the fornix or tarsus. We report a case of a patient presenting with a solitary compound cystic nevus of the conjunctival fornix in the background of bilateral complexion-associated melanosis. Case Presentation: A 71-year-old African-American female was referred for evaluation of an incidentally noted melanocytic lesion of the right conjunctival fornix. The patient underwent an excisional biopsy, revealing histological features consistent with a compound cystic nevus. Conclusion: This finding is noteworthy due to the rarity of conjunctival nevi originating in the fornix. The case underscores the importance of excisional biopsy in evaluating conjunctival forniceal melanocytic lesions to exclude malignant melanoma, a critical consideration for prognosis.

Coexisting Biopsy-Diagnosed Dementia and Glioblastoma.

Both glioblastoma (GBM) and dementia are devastating diseases with limited treatments that are usually not curative. Having clinically diagnosed dementia with an associated biopsy-proven etiology and a coexisting GBM diagnosis is a rare occurrence. The relationship between the development of neurodegenerative dementia and GBM is unclear, as there are conflicting reports in the literature. We present two cases of simultaneous biopsy-proven dementia, one with Alzheimer's disease (AD) and GBM, and one with cerebral amyloid angiopathy (CAA) and GBM. We discuss how these diseases may be associated. Whether one pathologic process begins first or develops concurrently is unknown, but certain molecular pathways of dementia and GBM appear directly related while others inversely related. Further investigations of these close molecular relationships between dementia and GBM could lead to development of improved diagnostic tools and therapeutic interventions for both diseases.

National pilot of entrustable professional activities in pathology residency training.

Entrustable professional activities (EPAs) are observable clinical skills and/or procedures that have been introduced into medical education at the student and resident levels in most specialties to determine readiness to advance into residency or independent practice, respectively. This publication describes the process and outcomes of a pilot study looking at the feasibility of using two anatomic pathology and two clinical pathology EPAs in pathology residency in 6 pathology residency programs that volunteered for the study. Faculty development on EPAs and their assessment was provided to pilot program faculty, and EPA assessment tools were developed and used by the pilot programs. Pre- and post-study surveys were given to participating residents, faculty, and program directors to gauge baseline practices and to gather feedback on the EPA implementation experience. Results demonstrated overall good feasibility in implementing EPAs. Faculty acceptance of EPAs varied and was less than that of program directors. Residents reported a significant increase in the frequency with which faculty provided formative assessments that included specific examples of performance and specific ways to improve, as well as increased frequency with which faculty provided summative assessments that included specific ways to improve. EPAs offered the most benefit in setting clear expectations for performance of each task, for providing more specific feedback to residents, and in increasing Program director's understanding of resident strengths abilities and weaknesses.

Adult primitive neuroectodermal tumors: the prognostic value of supratentorial location.

Primitive neuroectodermal tumors (PNETs) are tumors which primarily consist of undifferentiated round neuroepithelial cells. Central nervous system PNETs can be divided into two genetically distinct groups: infratentorial PNET (iPNET)/medulloblastoma and supratentorial PNET (sPNET). Currently, the comparative outcome of adult patients with sPNETs and iPNETs is unknown. In this study we have utilized the Surveillance, Epidemiology, and End Results database to perform a comparative analysis of 103 cases of adult sPNET and 669 adult medulloblastoma cases. Additionally we have analyzed various factors to identify their prognostic significance and characterize the optimal treatment for these tumors. Patients with sPNETs were seen to have a significantly worse survival than those diagnosed with medulloblastomas (16 vs. 155 months, p < 0.0001). Elderly patients (15 vs. 114 months, p < 0.0001) and those over the age of 40 (68 vs. 147 months, p < 0.0001) experienced significantly worse survival than younger patients. In contrast, radiotherapy (143 vs. 26 months, p < 0.0001), surgical resection (116 vs. 22 months, p = 0.0010) and the extent of resection (EOR) (173 vs. 81 months, p = 0.0005) resulted in significantly improved patient survival. Multivariate analysis revealed age greater than 40 years (HR: 1.57; 95 % CI: 1.17-2.11; p = 0.0028) and sPNET pathology (HR: 3.41; 95 % CI: 2.47-4.72; p < 0.0001) to be poor prognostic factors for survival while radiotherapy (HR: 0.52; 95 % CI: 0.38-0.71; p < 0.0001) and the EOR (HR: 0.73; 95 % CI: 0.55-0.96; p = 0.023) were associated with significantly improved survival. The treatment of sPNETs should therefore include maximal surgical resection when feasible followed by radiotherapy as these treatments have been demonstrated to confer a survival benefit. Additional studies are needed to identify effective chemotherapeutics and specific treatment regimens for adults with sPNETs.

Rapidly regulating platelet activity in vivo with an antidote controlled platelet inhibitor.

Millions of individuals are prescribed platelet inhibitors, such as aspirin and clopidogrel, to reduce their risk of thrombosis-related clinical events. Unfortunately many platelet inhibitors are contraindicated in surgical settings because of their inherent bleeding risk complicating the treatment of patients who require surgery. We describe the development of a potent antiplatelet agent, an RNA aptamer-termed Ch-9.14-T10 that binds von Willebrand factor (VWF) with high affinity and inhibits thrombosis in a murine carotid artery damage model. As expected, when this potent antiplatelet agent is administered, it greatly increases bleeding from animals that are surgically challenged. To improve this antiplatelet agent's safety profile, we describe the generation of antidotes that can rapidly reverse the activity of Ch-9.14-T10 and limit blood loss from surgically challenged animals. Our work represents the first antidote controllable antiplatelet agent, which could conceivably lead to improved medical management of patients requiring antiplatelet medication who also need surgery.

A Plant Stem Inoculation Assay for Assessing Transmission of Phytophthora infestans from Potato Seed Tubers to Emerged Shoots.

A stem inoculation assay was developed to assess transmission of Phytophthora infestans from the base of belowground potato stems to emerged aboveground shoots. Sporangia of P. infestans subsequently developed on aboveground stems after belowground stems were inoculated with isolates BF05 (US 8 genotype), WA10.1 (US 24), and 110B (US 11) by (i) placing a filter paper square saturated with a spore suspension onto nonwounded belowground stem tissue, (ii) placing a filter paper square saturated with a spore suspension onto wounded belowground stem tissue, and (iii) dipping the base of the cut belowground stem into a spore suspension. Plant stems were then incubated in a humidity chamber at 10 to 21.5°C for 7 to 28 days. Incidence of aboveground stems with sporulation was significantly less when stems were inoculated with isolate 110B than with isolates BF05 and WA10.1 but length of latent period did not differ among the three isolates. Incidence of aboveground stems with sporangia was significantly (P < 0.05) greater for 'Russet Norkotah' than for 'Umatilla Russet' when inoculated near the point of seed piece attachment. Incidence of aboveground stems with sporulation did not differ among incubation temperatures of 10, 15, and 21.5°C. Latent period significantly increased as temperature decreased. Age of stem did not appear to affect incidence of transmission. Incidence of transmission of P. infestans to aboveground stems from the base of belowground stems was significantly lowered when aboveground stems were treated with selected fungicides 1 h before inoculation.