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Loss of GBP1 expression does not impact PSaRC318 cell migration.

Prevalence of somatic DNA damage repair gene variants in Ewing cell lines included in original studies of PARP inhibition.

Ewing sarcoma is a fusion oncoprotein–driven primary bone tumor. A subset of patients (∼10%) with Ewing sarcoma are known to harbor germline variants in a growing number of genes involved in DNA damage repair. We recently reported our discovery of a germline mutation in the DNA damage repair protein BARD1 (BRCA1-associated RING domain-1) in a patient with Ewing sarcoma. BARD1 is recruited to the site of DNA double stranded breaks via the PARP protein and plays a critical role in DNA damage response pathways including homologous recombination. We thus questioned the impact of BARD1 loss on Ewing cell sensitivity to DNA damage and the Ewing sarcoma transcriptome. We demonstrate that PSaRC318 cells, a novel patient-derived cell line harboring a pathogenic BARD1 variant, are sensitive to PARP inhibition and by testing the effect of BARD1 depletion in additional Ewing sarcoma cell lines, we confirm that BARD1 loss enhances cell sensitivity to PARP inhibition plus radiation. In addition, RNA-sequencing analysis revealed that loss of BARD1 results in the upregulation of GBP1 (guanylate-binding protein 1), a protein whose expression is associated with variable response to therapy depending on the adult carcinoma subtype examined. Here, we demonstrate that GBP1 contributes to the enhanced sensitivity of BARD1-deficient Ewing cells to DNA damage. Together, our findings demonstrate the impact of loss-of function mutations in DNA damage repair genes, such as BARD1, on Ewing sarcoma treatment response.Significance:This work provides preclinical support for the inclusion of pediatric patients with advanced Ewing sarcoma and pathogenic germline variants in BARD1 in future clinical trials testing novel agents inducing DNA damage/targeting DNA damage repair.

Milky spots of the omentum enable lymphocyte access to the peritoneal cavity. In this issue of JEM, Yoshihara and Okabe (2023. J. Exp. Med.https://doi.org/10.1084/jem.20221813) demonstrate how secretion of retinoic acid by fibroblastic reticular cells allows lymphocyte entry into milky spots and the peritoneal cavity.

Ewing sarcoma is a fusion-oncoprotein-driven primary bone tumor most commonly diagnosed in adolescents. Given the continued poor outcomes for patients with metastatic and relapsed Ewing sarcoma, testing innovative therapeutic approaches is essential. Ewing sarcoma has been categorized as a ‘BRCAness’ tumor with emerging data characterizing a spectrum of DNA damage repair defects within individual Ewing tumors, including the presence of EWSR1::FLI1 itself, recurrent somatic mutations, and rare germline-based defects. It is critical to understand the cumulative impact of various DNA damage repair defects on an individual Ewing tumor’s response to therapy. Further, in addition to DNA-damage-directed therapies, subsets of Ewing tumors may be more susceptible to DNA-damage/immunotherapy combinations given the significant cross-talk between DNA damage and inflammatory pathways in the tumor microenvironment. Here we review potential approaches utilizing DNA-damaging agents as modulators of the Ewing tumor immune microenvironment, with a focus on radiation and opportunities during disease metastasis and relapse.

This systematic review and meta-analysis explores the effectiveness of teacher interventions supporting children with externalizing behaviors based on teacher and child outcomes. A systematic search was conducted using 5 electronic databases. From 5714 papers, 31 papers that included interventions delivered directly to teachers and aimed to benefit either teachers and/or children with externalizing behaviors were included. The review focused on qualified teachers working with children aged 2–13. The results of the current meta-analysis revealed a positive effect of teacher intervention on teacher and child outcomes, including the increased use of teacher-appropriate strategies, as well as significant and moderate improvements in teacher–child closeness, and small reductions in teacher–child conflict. For child outcomes, the interventions reduced externalizing behavior problems and ADHD symptoms and enhanced prosocial behavior. Only one fully blinded analysis for conduct problems was possible and revealed a moderate but significant reduction in favor of intervention. These findings provide evidence to support the role of teacher interventions for both teachers and children with externalizing behaviors. Future research should include more PBLIND measurements so that MPROX findings can be confirmed. More research should be done to evaluate the influence of teacher interventions on teachers’ well-being.

Familial hemophagocytic lymphohistiocytosis (HLH) is a life-threatening hyperinflammatory syndrome resulting from defective cytotoxicity. A previously healthy 3-month-old female presented with fever, irritability, abdominal distention, and tachypnea. She ultimately met all eight HLH-2004 diagnostic criteria, accompanied by elevated CXCL9. Initial empiric anti-inflammatory treatment included anakinra and IVIg, which stabilized ferritin and cytopenias. She had molecular and genetic confirmation of perforin deficiency and was started on dexamethasone and etoposide per HLH-94. She clinically improved, though CXCL9 and sIL-2Ra remained elevated. She was readmitted at week 8 for relapsed HLH without clear trigger and HLH-94 induction therapy was reinitiated. Her systemic HLH symptoms failed to respond and she soon developed symptomatic CNS HLH. She was incidentally found to have multifocal lung and kidney nodules, which were sterile and consisted largely of histiocytes and activated, oligoclonal CD8 T cells. The patient had a laboratory response to salvage therapy with alemtuzumab and emapalumab, but progressive neurologic decline led to withdrawal of care. This report highlights HLH foci manifest as pulmonary/renal nodules, demonstrates the utility of monitoring an array of HLH biomarkers, and suggests possible benefit of earlier salvage therapy.

Background: Ewing sarcoma is a rare, aggressive, fusion oncoprotein-driven pediatric cancer. Patients with upfront metastatic or surgically unresectable disease commonly undergo radiation as part of standard of care therapy. Currently, little is known about the effect of radiation specifically on the immune microenvironment of Ewing tumors, as no routine biopsies or other clinical samples are acquired during radiation. Further, historically, the field has lacked an immunocompetent mouse model of Ewing sarcoma in which to study Ewing tumor-immune cell interactions. Given our interest in understanding the anti-tumor immune response specifically during times of DNA damage, we sought to model Ewing sarcoma in a humanized mouse model (where human immune cell interactions with human tumor cells can be examined) in order to examine the Ewing tumor immune microenvironment (TIME) during radiation. Here, we determine the likeness of our established humanized mouse model of Ewing sarcoma to that of human Ewing tumors at baseline and examine changes in the Ewing TIME upon delivery of radiation. Methods: Blood from humanized NSG mice was analyzed by flow cytometry to confirm human immune cell reconstitution. Human Ewing tumor cells were injected into humanized mice and allowed to grow for approximately three weeks. Tumors were treated with radiation (either single dose or fractioned doses x 5 days) using the MultiRad350 Precision irradiator and lead shields were used to isolate the tumor area in the radiation field. Immune cells infiltrating tumors with/without radiation exposure were subsequently analyzed by multiplexed immunohistochemistry, flow cytometry, PCR, and RNAseq analysis. Ewing tumor immune infiltrates (as baseline/without radiation) from humanized mice models were compared to that of patient Ewing tumors. Results: Ewing tumors from both established and primary cell lines were successfully established in humanized mice. Similar to patient primary Ewing tumor samples, local Ewing tumors in humanized mice demonstrate a proportion of T-cell infiltration, although the overall number of infiltrating immune cells is low. Macrophage populations in Ewing sarcoma are not as predominant as in tumors such as osteosarcoma and are represented in this model. Analyses detailing the spatial localization and transcriptional profiles of Ewing tumor immune infiltrates following radiation are ongoing. Conclusions: In the absence of syngeneic and transgenic models of Ewing sarcoma, the use of humanized mouse models is a feasible alternative to address specific questions regarding the Ewing sarcoma TIME. Understanding the Ewing immune microenvironment during radiation therapy, a commonly used treatment modality in Ewing sarcoma, provides clues as to promising agents that may be worthy of preclinical testing to enhance the anti-tumor immune response during radiation in high-risk patients. Citation Format: Jessica D. Daley, Elina M. Mukherjee, Anthony R. Cillo, Adriana C. Tufino, Nathanael G. Bailey, Tullia C. Bruno, Linda M. McAllister-Lucas, Dario A. Vignali, Kelly M. Bailey. Radiation-induced changes to the immune microenvironment in an immunocompetent mouse model of Ewing sarcoma [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr A004.

Significance: Real-time information about oxygen delivery to the hepatic graft is important to direct care and diagnose vascular compromise in the immediate post-transplant period. Aim: The current study was designed to determine the utility of visible diffuse reflectance spectroscopy (vis-DRS) for measuring liver tissue saturation in vivo. Approach: A custom-built vis-DRS probe was calibrated using phantoms with hemoglobin (Hb) and polystyrene microspheres. Ex vivo (extracorporeal circulation) and in vivo protocols were used in a swine model (n = 15) with validation via blood gas analysis. Results:In vivo absorption and scattering measured by vis-DRS with and without biliverdin correction correlated closely between analyses. Lin’s concordance correlation coefficients are 0.991 for μa and 0.959 for μs ′ . Hb measured by blood test and vis-DRS with (R2 = 0.81) and without (R2 = 0.85) biliverdin correction were compared. Vis-DRS data obtained from the ex vivo protocol plotted against the PO2 derived from blood gas analysis showed a good fit for a Hill coefficient of 1.67 and P50 = 34 mmHg (R2 = 0.81). A conversion formula was developed to account for the systematic deviation, which resulted in a goodness-of-fit R2 = 0.76 with the expected oxygen dissociation curve. Conclusions: We show that vis-DRS allows for real-time measurement of liver tissue saturation, an indicator for liver perfusion and oxygen delivery.

Ewing sarcoma, an oncofusion-driven primary bone tumor, can occur in the setting of various germline mutations in DNA damage repair pathway genes. We recently reported our discovery of a germline mutation in the DNA damage repair protein BARD1 (BRCA1-associated RING domain-1) in a patient with Ewing sarcoma. BARD1 is recruited to the site of DNA double stranded breaks via the poly(ADP-ribose) polymerase (PARP) protein and plays a critical role in DNA damage response pathways including homologous recombination. PARP inhibitors (PARPi) are effective against Ewing sarcoma cells in vitro, though have demonstrated limited success in clinical trials to date. In order to assess the impact of BARD1 loss on Ewing sarcoma sensitivity to PARP inhibitor therapy, we generated the novel PSaRC318 patient-derived Ewing tumor cell from our patient with a germline BARD1 mutation and then analyzed the response of these cells to PARPi. We demonstrate that PSaRC318 cells are sensitive to PARP inhibition and by testing the effect of BARD1 depletion in additional Ewing sarcoma cell lines, we confirm that loss of BARD1 enhances PARPi sensitivity. In certain malignancies, DNA damage can activate the IRF1 (interferon response factor 1) immunoregulatory pathway, and the activation of this pathway can drive immunosuppression through upregulation of the immune checkpoint protein PD-L1. In order to determine the ability of PARPi to alter Ewing tumor immunoregulation, we evaluated whether PARPi results in upregulation of the IRF1-PDL1 pathway. Indeed, we now demonstrate that PARPi leads to increased PD-L1 expression in Ewing sarcoma. Together, these data thus far suggest that while Ewing tumors harboring germline mutations in DNA damage repair proteins may in respond to PARPi in vitro, in vivo benefit of PARPi may only be demonstrated when counteracting the immunosuppressive effects of DNA damage by concurrently targeting immune checkpoint proteins.

Introduction: Because it is impossible for two separate machined surfaces to be microscopically identical, a problem can occur if firearm and toolmark examiners are unable to base an identification of weapons and other related components derived from a crime scene. The presence of small parts used in firearms usually produces marks useful in the identification of cartridge cases or bullets that was recently discharged from a firearm. There have been doubts regarding the uniqueness of these and other marks left on the fired evidence obtained due to a lack of characteristic markings (Bonfanti and De Kinder 3-10). Reproducing marks are more liable to happen due to the weapon’s manufacture process. As such, manufacturing processes needs to be investigated as reproducing marks can vary from time to time. Method: After obtaining and prepping the Metal Injection Molded extractors and ammunition as well as the Progressive Die Stamped extractors and ammunition, ten rounds per extractor were fired from a Remington Shotgun and Hi-Point Firearm, respectively. Comparisons for the identification of extractor markings were conducted on the ammunition. Results: Analysis of the Metal Injection Molded extractors concluded that all the Metal Injection Molded extractors were created after being injected in the same mold cavity. After injection of the feedstock into the mold, the produced part was either removed or broken off the mold. This would help to explain why there was no sign of reproducing areas on the inside working surfaces of the extractors due to a grinding or polishing finishing procedure. Reproducing markings were, however, displayed on other areas of the extractor. The markings produced by these extractors were less profound and was determined that this was due to the cartridge contacting the extractor with enough force, but not with as much force as with the Progressive Die Stamped extractor. Further investigation of the Progressive Die Stamped extractors revealed markings identified as markings obtained at the conclusion of the manufacturing process. Additional markings were also seen and was identified as those derived from a sandpaper. The comparison of extractor markings seen within the Progressive Die Stamped extractors revealed significant agreement of the overall pattern among each group of extractors. The markings produced by these extractors were more profound and it was determined that this was due to the cartridge contacting the extractor with much force. When comparing markings from various extractor groups, it was found that more differences than similarities existed among the markings. Although there were more similarities among the Metal Injection Molded markings produced by different extractors when conducting an inner comparison, these markings can statistically and microscopically be differentiated. Conclusion: Although microscopic comparisons produced similarities among the extractor markings from both manufacturing processes, mass produced parts by modern manufacturing processes; Metal Injection Molding and Progressive Die Stamping, can differentiated by firearm examiners after comparing fired evidence.

BackgroundPediatric patients with metastatic or recurrent bone sarcomas have poor survival, necessitating new therapies. Studies of immunobiology in pediatric bone sarcomas have focused on analysis of samples from surgical local control, when patients are receiving immunosuppressive chemotherapy, with little data available from relapse. Here, we sought to leverage transcriptomic and imaging approaches in tandem to characterize the immune landscape of primary and recurrent Ewing sarcoma and osteosarcoma and to identify new therapeutic avenues for these patient populations.MethodsSingle-cell RNAseq (scRNAseq; 10X Genomics) was performed on sorted CD45+ cells from paired peripheral mononuclear cells (PBMC) and tumor infiltrating leukocytes (TIL) from freshly resected bone tumor specimens in the setting of pre-treatment or >6 months post-chemotherapy in the setting of disease relapse. Multiplexed immunofluorescence (mIF) analysis for CD45, DAPI, CD4, CD8, CD68, CD20, and FOXP3 was also performed on FFPE tissue from the same tumors. CIBERSORTx was used in conjunction with TARGET-OS bulk RNAseq from osteosarcoma tumors to infer cell type frequencies. Expression of ligands and receptors in primary versus relapsed disease was assessed from scRNAseq using CellTalker.ResultsWe analyzed a total of 29,993 cells from 20 donors (4 paired PBMC/TIL from osteosarcoma, 4 paired samples from Ewing sarcoma, 4 healthy donor PBMC) by scRNAseq. A total of 5 TIL samples were from primary disease sites and 3 were from metastatic sites. We identified major immune populations by canonical expression profiles and used transcriptional profiles from TIL to derive a signature matrix for CIBERSORTx. In 88 osteosarcoma samples from TARGET-OS, we found that higher frequencies of CD14+CD16+ macrophages were associated with better survival (HR:0.28, p = 0.01). scRNAseq from our cohort revealed expression of CXCL12, CCL7, and CCL3L1 by CD14+CD16+ macrophages, suggesting this macrophage population may drive tumor immune infiltration. In both Ewing sarcoma and osteosarcoma, mIF revealed greater numbers of tumor-infiltrating immune cells in the setting of relapse versus primary tumors. scRNAseq analysis revealed higher levels of interferon-gamma expressing CD8+ T cells and CD4+ regulatory T cells in relapsed versus primary disease, suggesting that recurrent tumors may be more immunogenic.ConclusionsAlthough pediatric bone sarcomas are typically considered “immunologically cold”, our transcriptomic and imaging approaches revealed a role for a myeloid cell subset in overall survival and increased immune infiltration and T cell activation in recurrent disease. These data suggest specific immunotherapeutic avenues should be tailored to both primary and recurrent disease to improve outcomes in pediatric bone sarcoma.Ethics ApprovalHuman specimens were collected with written informed consent under the IRB approved STUDY19030108 and the IRB approved Musculoskeletal Oncology Biobank and Tumor Registry.

e21576 Background: Melanoma is the dominant skin cancer of pediatric patients, and accounts for 1-3% of pediatric malignancies. The prognosis for both adults and children with melanoma correlates with stage at diagnosis, and initial Breslow depth is a critical determinant. Kaplan-Meier melanoma-specific survival curves show survival rates ranging from 98% (T1a) to 75% (T4b) at 10 years for adult patients with stage I and II disease at diagnosis. Pediatric patients are known to present with thicker primary melanomas as compared to their adult counterparts. The preferred biopsy method of the American Academy is Dermatology for melanoma is excisional, however partial shave biopsy is most frequently performed in children, given its simplicity, efficiency, and often low clinical suspicion for cutaneous malignancy. Despite the ease of this biopsy technique, shave biopsy has a high rate of base transection, reducing accuracy of microstaging, which is crucial for therapy planning. Methods: Retrospective chart review evaluation of the potential effects of biopsy method on staging, surgical recommendations, and treatment approach for pediatric patients with melanocytic tumors. Results: Data was available for 91 pediatric patients with a spectrum of melanocytic tumors ranging from atypical with unknown malignant potential, to melanoma. Patient characteristics including gender, age at biopsy, biopsy method, status of biopsy margins, recommendations for re-excision, sentinel lymph node biopsy, and treatment plan were collected and analyzed. There were 48 females and 43 males with age range 1—22 years (mean 10 years). Sixty-eight of 91 (75%) tumors had a positive margin (deep, peripheral, or both) on diagnostic biopsy, 52/68 (76.5%) were obtained using shave method. Evaluating the deep margin specifically, 50/91 (55%) tumors had a positive deep margin, 46/50 (92%) of which were obtained using shave method. No tumor evaluated by punch had a positive deep margin (0/18). Of all shave biopsies, 77% had positive deep margin as opposed to only 13% using alternative biopsy methods. In 10/91 (11%) patients, surgical recommendations were changed based on inaccurate microstaging and positive deep biopsy margins, 9/10 (90%) of these patients had undergone shave biopsy. Most (8/10) patients had melanoma, the remaining 2/10 had highly atypical spitzoid tumors in which melanoma could not be ruled out. Conclusions: Pediatric patients with melanocytic tumors most commonly underwent shave biopsy as the initial diagnostic biopsy method. This method of biopsy was associated with higher incidence of positive margins at diagnosis and more aggressive definitive surgical management. These findings suggest that more routine use of excisional biopsy in pediatric patients with lesions being evaluated for malignancy could decrease the incidence of inaccurate microstaging and reduce the need for more aggressive definitive surgical management.

In recent years, grid modernization has become vital to achieve a balance between energy economics and consumer comfort. As the cost of distributed energy resources (DERs) such as photovoltaics (PV) has declined, utility companies have become increasingly interested in incorporating these technologies into the smart grid. Even though the advantages from DER integration outweigh the challenges, its implementation is easier said than done. To overcome these issues, a grid apparatus with advanced monitoring and dynamic control capabilities must be efficiently commissioned. In turn, this enables reliable operation of a high-performance, economically viable power grid. This study utilizes an IEEE 34 bus system modified to integrate high penetration DERs alongside dynamically controllable voltage control devices (Eaton’s Voltage Regulator controller) to provide a broad understanding of the system reliability and resiliency. The three advanced apparatus operating control modes showcased are Sequential, Time-integrating and Voltage-averaging. In summary, this paper provides a comprehensive review of the commonly established voltage control modes and configurations in a voltage regulator as well as recommendations for enhancing performance of the system using various degrees of control.

Objective Osteopontin (OPN) plays a role in tumor progression. This study aimed to determine the expression of OPN, CD44, and integrin αvβ3 in pleomorphic adenoma (PA), acinic cell adenocarcinoma (ACA), and mucoepidermoid carcinoma (MEC). Study Design Immunohistochemistry was used to semiquantify the levels of expression of OPN and its receptors in normal salivary glands (NSG) (n = 20), PA (n = 20), ACA (n = 11), and MEC (n = 29). Results OPN expression was increased in ACA and MEC compared with PA and NSG (median scores, 6, 6, 4, and 4, respectively). CD44 expression was increased in ACA and reduced in MEC and PA compared with NSG (median scores, 8, 4, 3, and 5, respectively). Integrin αvβ3 median scores were 5 in ACA, 1 in MEC, and 0 in PA and NSG. Conclusions OPN is expressed in salivary gland tumors and is at higher levels in ACA and MEC.

Objectives Osteopontin (OPN) plays a role in tumor progression. This study aimed to determine the expression of OPN, CD44, and integrin αvβ3 in pleomorphic adenoma (PA), polymorphous low-grade adenocarcinoma (PLGA), and adenoid cystic carcinoma (ACC). Study Design Immunohistochemistry was used to semiquantify the level of expression of OPN and its receptors in normal salivary glands (NSG; n = 20), PA (n = 20), PLGA (n = 16), and ACC (n = 22). Results OPN expression was increased in PLGA and intermediate-/high-grade ACC compared with PA and NSG (median scores, 6, 5, 4, and 4, respectively). CD44 expression was reduced in PA, PLGA, and ACC. OPN expression levels were moderately correlated with CD44 in PLGA. Integrin αvβ3 was not expressed in PA and ACC and was seen in only 1 case of PLGA. Conclusions OPN is expressed in salivary gland tumors but does not correlate well with CD44 and αvβ3.

Objectives Kallikrein-related peptidase 10 (KLK10) has been implicated in the development of several types of cancer. The purpose of this study was to analyze the expression of KLK10 in 3 types of salivary gland tumour and normal salivary glands. Materials and methods: A standard immunoperoxidase staining technique was used to assess the Immunoexpression profile of KLK10 in normal salivary glands and 3 types of salivary gland tumour: pleomorphic adenoma, adenoid cystic carcinoma and mucoepidermoid carcinoma. Results Pleomorphic adenomas showed significantly lower KLK10 levels than control tissues. Neither of the malignant tumours (adenoid cystic carcinoma and mucoepidermoid carcinoma) showed a significant alteration in the immunoreactive scores of KLK10 in comparison with the normal salivary gland tissues. KLK10 immunoreactive scores were comparable in adenoid cystic carcinoma and mucoepidermoid carcinoma. Pleomorphic adenoma had significantly lower levels of KLK10 than mucoepidermoid carcinoma. Conclusions The finding of lower KLK10 levels in pleomorphic adenoma suggests aberrant expression in a tumour that develops primarily from myoepithelial cells. A kallikrein cascade may play a role in the development and/or outcome of some salivary gland tumours.

Objective The purpose of the study was to present the clinical and microscopic features of oral foreign body granulomas associated with the injectable dermal filler Radiesse, and to increase awareness of this potential complication as the use of dermal fillers by the aging Baby Boom generation increases. Study Design Demographic and clinical data of 8 patients with foreign body granulomas containing the characteristic microscopic appearance of hydroxyapatite microspheres were studied. Analysis of the microspheres of 1 case was carried out by energy dispersive x-ray microanalysis to confirm the calcium and phosphorus content. Results The clinical and microscopic appearance of Radiesse-induced foreign body granulomas is illustrated and discussed. Conclusions Foreign body granulomas containing hydroxyapatite microspheres in patients receiving treatment with the cosmetic filler Radiesse have a characteristic microscopic appearance. Oral Radiesse-induced nodules occur more often in older women, most commonly in the lips and the mandibular labial vestibule.

c d l In 1987, Miles et al described 2 otherwise healthy patients, 3.5 and 15 years of age, who exhibited congenital mild facial asymmetry and unilateral enlargement of the maxillary gingiva and alveolar bone. One patient had hypoplastic teeth in the affected quadrant and missing premolars, and the other patient had hypoplasia of only the premolars. The older patient also exhibited hypertrichosis of skin in the affected area. The investigators suggested the term hemimaxillofacial dysplasia (HD) for this condition. In 1990, Danforth et al decribed 8 patients with similar features, but without nown skin changes, and proposed the term segental odontomaxillary dysplasia (SOD), which hey believed to be a more accurate descriptor. In 996, DeSalvo et al reported a case involving a 7-year-old girl, using the term SOD, associated with an area of hypopigmentation of the upper lip on the affected side. Also, in 1996, Packota et al detailed the maxillary radiographic features of 12 cases of SOD, including bony sclerosis with thickened vertical trabeculae in the affected area, missing premolars, delayed eruption of adjacent teeth, and a small maxillary sinus on the affected side. In 2004, Welsch and Stein reported on 1 patient with Becker’s evus of the skin and recommended the acronym ATS (hemimaxillary enlargement, asymmetry of he face, tooth abnormalities, and skin findings) to dd yet another term for this condition. Other cases

We compared the development of microalgal and bacterial-denitrifier communities within biofilms over 28 days in a restored-prairie stream (RP) and a stream receiving treated wastewater effluent (DER). Inorganic nutrient concentrations were an order of magnitude greater in DER, and stream waters differed in the quality of dissolved organics (characterized via pyrolysis-GC/MS). Biofilm biomass and the densities of algae and bacteria increased over time in both systems; however, algal and denitrifier community composition and the patterns of development differed between systems. Specifically, algal and denitrifier taxonomic composition stabilized more quickly in DER than RP, whereas the rates of algal and denitrifier succession were more closely coupled in RP than DER. We hypothesize that, under unenriched conditions, successional changes in algal assemblages influence bacterial denitrifiers due to their dependence on algal exudates, while under enriched conditions, this relationship is decoupled. Between-system differences in organic signatures supported this, as RP biofilms contained more labile, aliphatic compounds than DER. In addition, potential denitrification rates (DNP) were negatively correlated with the percentage of aromatic compounds within the biofilm organic signatures, suggesting a significant relationship between algal exudate composition and denitrification. These results are significant because anthropogenic factors that affect biofilm community composition may alter their capacity to perform critical ecosystem services.

The incidence of hepatocellular carcinoma has shown an alarming increase in North America and Europe since 1980, as has the incidence of reported metastases to the oral region. Consequently, it is important that head and neck health care specialists be familiar with recent epidemiological trends and the histologic and recent immunohistochemical advances in the diagnosis of this deadly malignancy. Histologic features are reviewed, and recent advances in the immunohistochemical differential diagnosis are detailed and illustrated in two cases. A panel of immunohistochemical stains is recommended, some or all of which may be useful to differentiate poorly differentiated or pseudoglandular hepatocellular carcinoma from other metastatic adenocarcinomas to the oral and maxillofacial region.

Recently identified as a key component of the murine periodontal ligament (PDL), periostin has been implicated in the regulation of collagen fibrillogenesis and fibroblast differentiation. We investigated whether periostin protein is expressed in the human PDL in situ and the mechanisms regulating periostin expression in PDL fibroblasts in vitro. With immunohistochemistry, periostin protein was identified in the PDL, with expression lower in teeth with reduced occlusal loading. In vitro application of uniaxial cyclic strain to PDL fibroblasts elevated periostin mRNA levels, depending on the age of the patient. Treatment with transforming growth factor-beta1 (TGF-β1) also significantly increased periostin mRNA levels, an effect attenuated by focal adhesion kinase (FAK) inhibition. FAK-null fibroblasts contained no detectable periostin mRNA, even after stimulation with cyclic strain. In conclusion, periostin protein is strongly expressed in the human PDL. In vitro, periostin mRNA levels are modulated by cyclic strain as well as TGF-β1 via FAK-dependent pathways.

The installation of independent-leg jack-up rigs in seabed sediments where a strong layer overlays weaker soil can lead to a catastrophic ‘punch-through’ event, with potential leg buckling or toppling of the unit. In an attempt to mitigate this risk in layered clays, an industry practice known as ‘perforation drilling’ is sometimes used. This process is also colloquially referred to as Swiss cheese drilling. The rationale is to reduce the bearing resistance of the upper stiff layer by perforating it with drill holes. Although drilling directly underneath the spudcan is reported to ease punch-through, little guidance to how many or where drill holes should be located is available. This paper reports two series of model tests of a 40 mm diameter ( D ) spudcan footing vertically installed in a stiff-over-soft clay deposit. In this preliminary study, the effectiveness of perforation drilling was investigated by drilling holes of different spacing, depth and distribution both underneath and outside the immediate perimeter of the penetrating spudcan. Drilling patterns were chosen to not just follow the conventional method of holes directly below the spudcan, but to coincide with failure mechanisms previously observed during spudcan penetrations in stiff-over-soft clay soils. The method of producing the holes, whether through drilling, coring or water jetting, was also investigated. Results show that punch-through can be mitigated when the layered deposit is punctured in a zone immediately outside the spudcan periphery.

Essentially, sialoblastoma is a disease of infancy with the oldest case presenting at 4 years of age. About one third of pediatric sialoblastoma cases will have a cribriform growth pattern. No adult cases have been reported with a specific diagnosis of sialoblastoma. If even focal cribriforming were present, such cases have undoubtedly been diagnosed as adenoid cystic carcinoma. Such was the circumstance in the 3 adult tumors presented in this report. Each case, however, has the primitive histopathology with discrete nests of basaloid tumor cells, associated bilayered ductal structures and the fibromyxoid stroma characteristic for sialoblastoma with its resemblance to fetal salivary gland or salivary gland with arrested development. One key example has 28-year follow-up. Sialoblastoma, whether in a child or adult with or without a cribriform growth pattern, appears to have a more favorable prognosis than adenoid cystic carcinoma. Aspects of the histological differential diagnosis of these 2 tumors are discussed.

CASE REPORT A 65-year-old male, referred for minimal otalgia and “pressure” sensations in the area of the left jaw, presented with a 25-mm-diameter radiolucent lesion located in a pericoronal relationship surrounding the crown of the impacted left mandibular third molar (Fig. 1). His medical history was significant for hypertension and hypercholesterolemia, for which he was taking bisoprolol, Lipitor, and acetylsalicylic acid. Radiographically, the lesion appeared to be unilocular and extended into the ascending ramus. Its margins were mostly well circumscribed, although a localized area of irregular bone destruction with indistinct margins was noted along the superior margin. The borders of the anterior margin were distinct. The posterior margin appeared to extend to the anterosuperior aspect of the inferior alveolar canal. No bony expansion was noted. The lesion was of unknown duration.

The increasing lateral lengths of wellbore and additional composite frac plugs (CFP) per well has created a need for a more efficient and aggressive post frac millout of plugs using coiled tubing (CT). Economic post job analysis has shown some advantages and drawbacks of using larger diameter coiled tubing and more robust tools, all while developing a better understanding of the impact to the operation. Several complexities exists downhole that are not well understood. Attempts to comprehend downhole conditions are limited by past experiences, pre-job software modeling, and post-job matching through the uses of surface parameter but not actual downhole data. Monitoring downhole forces applied when performing post frac milling operations, through the use of a memory sub, gives a better understanding of forces that are actually transmitted from the tools to the CFP in the actual wellbore environment. Thru tubing tools selection is based on actual downhole data which translates to an overall increase in efficiency of the operation. A closer look at the actual downhole parameters, when performing operations with larger diameter CT and extend reach tools, lead to findings that the weight on bit (WOB) applied to a plug in the lateral of a well can be as high as 15,000 lbf. This amount of force far exceeds what is applied at surface and what conventional software modeling shows. This also greatly surpasses the recommended WOB while drilling the CFP. Applying substantial downward force to the CFP results in movement of the plug prior to being completely milled up, which causes significant operational issues. These same forces that cause the aforementioned issues are necessary to aid in having consistent mill times throughout the operation, enough WOB to get CT to reach measure depth (MD), and produce an operation that is completed in a timely and economically sound manner.

The human kallikrein 13 protein (hK13) is expressed in many normal tissues. Petraki et al have previously described presence of hK13 in salivary gland tissue, localized to duct epithelia and some acinar cells. The aim of this study was to determine whether hK13 is expressed in salivary gland tissues and salivary gland tumors (both benign and malignant), in order to compare normal with tumor tissues. Pleomorphic adenomas (PA), adenoid cystic carcinomas (ACC), polymorphous low grade adenocarcinomas (PLGA), acinic cell carcinomas (ACI), mucoepidermoid carcinomas (MEC) and adenocarcinomas not otherwise specified (ANOS) of both minor and major salivary glands were examined. The results of this study indicate that most salivary gland tumors show high levels of expression of hK13. Overall, staining in PA was significantly less than that seen in normal salivary gland tissue. PLGA, ACC and ANOS each stained significantly more than normal salivary gland tissue while MEC and ACI did not. Ductal cells and cells lining duct-like structures showed a higher intensity of staining than non-ductal cells in most tumors. Tumors which exhibited only non-ductal cells also exhibited cytoplasmic staining. In conclusion, we demonstrate the high expression of hK13 in several common salivary gland tumors.

Osteopontin (OPN) is expressed in numerous carcinomas and plays a role in tumour development, invasion and metastasis. This study examines by immunohistochemistry the expression of OPN in normal salivary gland tissue and three types of salivary gland tumour: pleomorphic adenoma (PA), adenoid cystic carcinoma (ACC) and polymorphous low grade adenocarcinoma (PLGA). PAs and PLGAs demonstrated higher levels of OPN than normal salivary gland tissue, while ACC, although showing a trend towards increased OPN, was not significantly different. The results of this study indicate that OPN expression is present in normal salivary gland tissue, and is increased in certain salivary gland tumours, but further investigation is necessary to clarify its role.

There has been considerable disagreement among educators on the topics and details of topics that should be included in the teaching of oral pathology to dental students and graduate students in dental specialties. Various authorities have recommended core curricula that range from comprehensive teaching of eighteen categories, each with up to nine subheadings, covering hundreds of entities, to as few as approximately fifty of the most common lesions that affect the oral and maxillofacial region. This article offers a curriculum planning model designed to help faculty make decisions about course content and emphases. The model allows instructors to assess content relevance and priority based on three criteria: 1) commonness, 2) uniqueness, and 3) significance of diseases and conditions. The product of this decision-making process is a relevance score that can serve as a guideline for the choice and details of topics to be included in oral pathology courses.

The human kallikrein 13 protein (hK13) is expressed in many normal tissues. Petraki et al have previously described presence of hK13 in salivary gland tissue, localized to duct epithelia and some acinar cells. The aim of this study was to determine whether hK13 is expressed in salivary gland tissues and salivary gland tumors (both benign and malignant), in order to compare normal with tumor tissues. Pleomorphic adenomas (PA), adenoid cystic carcinomas (ACC), polymorphous low grade adenocarcinomas (PLGA), acinic cell carcinomas (ACI), mucoepidermoid carcinomas (MEC) and adenocarcinomas not otherwise specified (ANOS) of both minor and major salivary glands were examined. The results of this study indicate that most salivary gland tumors show high levels of expression of hK13. Overall, staining in PA was significantly less than that seen in normal salivary gland tissue. PLGA, ACC and ANOS each stained significantly more than normal salivary gland tissue while MEC and ACI did not. Ductal cells and cells lining duct-like structures showed a higher intensity of staining than non-ductal cells in most tumors. Tumors which exhibited only non-ductal cells also exhibited cytoplasmic staining. In conclusion, we demonstrate the high expression of hK13 in several common salivary gland tumors.

The human kallikrein 5 protein (hK5) is expressed in many normal tissues, most notably in skin, breast, salivary gland and esophagus. It has also been shown to be a potential biomarker for breast, ovarian and testicular cancer. Human kallikrein 3 (hK3; prostate-specific antigen) is the most useful marker for adenocarcinoma of the prostate gland. The aim of this study was to determine whether hK3 and hK5 are expressed in salivary gland tissues and salivary gland tumors (both benign and malignant), in order to compare normal with tumor tissues. Pleomorphic adenomas, adenoid cystic carcinomas, polymorphous low-grade adenocarcinomas, acinic cell carcinomas, mucoepidermoid carcinomas and adenocarcinomas not otherwise specified of both minor and major salivary glands were examined. The results of this study indicate that most salivary gland tumors do not show high levels of expression of hK5. Staining was most prominent in keratinizing epithelia in pleomorphic adenomas. hK3 is not expressed in salivary gland tumors.

Bronchogenic cysts are lesions that are an anomaly of congenital development. Although most are found in the mediastinum, bronchogenic cysts can be found in many different anatomical positions in the body and can have multiple presentations. These lesions can be asymptomatic and found incidentally, or they can present with specific symptoms. Bronchogenic cysts can be diagnosed on multiple radiographic modalities including conventional radiography, computed tomography, and magnetic resonance imaging. Most are treated conservatively but the possibility of infection or malignant transformation has caused some clinicians to recommend invasive treatment for these lesions. This article presents a case report of the diagnosis and management of a paravertebral bronchogenic cyst in a 64-year-old female.

Two cases of osteosarcoma of the jaws in children are reported. One patient was a 13-year-old girl whose first symptoms included nasal and maxillary sinus congestion, followed by epistaxis. She was found to have chondroblastic osteosarcoma extending through the left maxillary alveolar process and sinus. Following surgery and chemotherapy, the patient has been free of disease for 7 years. The second patient, an 8-year-old boy, was diagnosed with juxtacortical (parosteal) osteosarcoma of the mandible, which is a less aggressive variant of the neoplasm. It is believed that this is the youngest patient reported to date with juxtacortical osteosarcoma of the jaws. He was treated by block resection of the right side of the mandible and is free of disease 3(1/2) years later.

Objective To investigate central lesions of the jaws dominated by foamy macrophages (foam cells), which are interpreted to be “central xanthomas of the jaws” and to differentiate this condition from reported cases of jaw nonossifying fibroma/fibrous cortical defect and benign fibrous histiocytoma of bone. Study Design The study is a literature review and a retrospective analysis of clinical, radiographic, microscopic, and immunohistochemical features of five new cases of this condition. Results The lesion has a predilection for the mandible in adults of variable ages, with the potential to become very large and destructive; however, it has a very low recurrence rate, if any, following surgical curettage. Conclusion We suggest that the “central xanthoma of the jaws” be considered a unique, although rare, clinicopathologic entity. It is unclear whether this lesion is a benign neoplastic process or a persistent reactive process.

Attention-deficit hyperactivity disorder (ADHD) and its associated behavioural manifestations develop and progress as the result of complex gene-environment interactions. Parents exert a substantial influence and play a major role in their child's social environment. Despite this, recent evidence has suggested that adapting the child's environment via parenting interventions has minimal effects on child ADHD symptoms when analysing data from informants who are probably blind to treatment allocation. However, adverse parenting and family environments may act as a source of environmental risk for a number of child outcomes beyond ADHD symptoms. This is a narrative review that critically discusses whether parenting interventions are beneficial for alternative functioning outcomes in ADHD including neuropsychological, academic and social functioning and disruptive behaviour and how parenting and familial environments may be associated with these outcomes. In addition, the review explores how parental depression and parenting efficacy impact on capacity for optimal parenting and whether parenting interventions benefit parents too. A review of the evidence suggests that with modification, parenting interventions are beneficial for a number of outcomes other than ADHD symptom reduction. Improving the parent-child relationship may have indirect benefits for disruptive behaviour. Furthermore, parenting behaviours may directly benefit child neuropsychological, academic and social functioning. Parenting interventions can have therapeutic benefits for parents as well as children, which is important as parent and child well-being is likely to have a transactional relationship. Evaluation of the clinical success of parenting interventions should focus on a wider range of outcomes in order to aid understanding of the multifaceted benefits that they may be able to offer. Parenting interventions should not be seen as a redundant adjunct to medication in multi-modal treatment approaches for ADHD; they have the potential to target outcomes that, at present, medication seems less able to improve.

Attention-deficit hyperactivity disorder (ADHD) is a complex disorder that can affect individuals across the lifespan. It is associated with substantial heterogeneity in terms of aetiology, clinical presentation and treatment outcome and is the subject of extensive research. Because of this, it can be difficult for clinicians to stay up to date with the most relevant findings and know how best to respond to parents' questions and concerns about the disorder and interventions. This is a narrative review that aims to summarize key findings from recent research into ADHD and its treatment that clinicians can share with families in order to increase their knowledge about ADHD and intervention options. ADHD develops as a result of complex interplay between interdependent genetic and non-genetic factors. The disorder is associated with substantial impairments in functioning and poor long-term outcomes. Pharmacological and non-pharmacological treatment options are available for symptom management and to improve function, but functioning outcomes often fail to normalize in children with ADHD. Despite extensive advances in understanding this complex disorder, it is clear that there is still a long way to go. In particular, we address the need for future non-pharmacological interventions to be more specifically targeted for ADHD symptoms and its commonly associated functioning deficits in order to ensure the best long-term outcomes for children with ADHD.

Amalgam tattoos occur when small particles of dental amalgam, composed largely of silver (Ag) and mercury (Hg), are inadvertently implanted into oral soft tissues during dental procedures. Metallothioneins (MTs) are ubiquitous, low molecular weight, cysteine-rich, metal-binding proteins that are inducible by many agents including metals and may be involved in the detoxification of toxic metals such as Hg. In this study, the correlation between MT expression and amalgam tattoos in human gingiva was investigated using energy-dispersive X-ray microanalysis (EDX) and immunohistochemical techniques. Light microscopically, amalgam tattoos presented as either fine granular particles or larger discrete opaque globular particles in connective tissues. EDX revealed the smaller particles to be silver sulphide (Ag(2)S), while the larger particles exhibited a shell of Ag(2)S that contained irregularly distributed masses of Ag and Hg. Particles of tin (Sn) were also found. No MT staining was observed in collagen, fibroblasts or blood vessels in areas exhibiting abundant amounts of embedded fine granular Ag(2)S particles. Blood vessels exhibiting relatively few amalgam particles stained positively for MT. Cells with the morphological features of histiocytes located directly adjacent to larger pieces of amalgam showed intense MT staining. These results indicate that amalgam tattoos contain no Hg or free Ag except in large globular pieces of amalgam, which still contain Hg and which induce MT expression in adjacent histiocytes. This suggests that Hg leaching from impacted dental amalgam particles induces MT, while residual Ag(2)S and Sn particles do not. MT may therefore act to reduce Hg exposure in patients with amalgam tattoos.

Acidic (FGF-1) and basic (FGF-2) fibroblast growth factors are members of a family of growth factors that function in growth, differentiation and regeneration of a variety of tissues. Their presence in human odontogenic cysts and tumors has not been previously investigated. This study was designed to detect immunohistochemically the presence of these factors and two fibroblast growth factor receptors (FGFR2 and FGFR3) in a cross section of odontogenic cysts and tumors, to determine if they may be involved in the differentiation of odontogenic epithelium or, more specifically, in the development of particular cysts or tumors. Archival formalin-fixed paraffin-embedded tissues were used. With some exceptions, FGF-2 and the receptor FGFR2, were found in the cytoplasm and occasionally in the nuclei of cells of odontogenic epithelium, while FGF-1 and the receptor FGFR3, were absent or only focally or weakly detected, using standard immunohistochemical techniques. The data are similar to those published for normal murine odontogenesis, suggesting that these factors are associated with odontogenic differentiation rather than pathogenesis. The presence of significant nuclear staining in odontogenic epithelium associated with embryonic mesenchyme in ameloblastic fibromas and ameloblastic fibro-odontomas suggests that FGF-2 may be involved in directing nuclear activity at the histodifferentiation stage of odontogenesis.

Oral lichen planus (OLP) is a common mucosal condition that is considered premalignant by some, whereas others argue that only lichenoid lesions with epithelial dysplasia are at risk of progressing into oral carcinoma. A recent study from this laboratory used microsatellite analysis to evaluate OLP for loss of heterozygosity (LOH) at loci on three chromosomal arms (3p, 9p, and 17p) (Am J Path 1997;Vol151:Page323-Page327). Loss on these arms is a common event in oral epithelial dysplasia and has been associated with risk of progression of oral leukoplakia to cancer. The data showed that, although dysplastic epithelium demonstrated a high frequency of LOH (40% for mild dysplasia), a significantly lower frequency of LOH was noted in OLP (6%), which is even lower than that in hyperplasia (14%). Such results do not support OLP as a lesion at risk for malignant transformation. As a second step of the research, we determined LOH frequencies in 61 dysplastic lichenoid lesions (mild 35; moderate 19; severe 7) using the same microsatellite markers and compared these results with data obtained from the first study and from 13 normal mucosal specimens. Dysplastic lichenoid lesions showed a high frequency of loss (54% for lichenoid lesions with mild dysplasia), but values did not differ significantly from those observed in dysplasia of similar degree without lichenoid appearance. None of the normal mucosa demonstrated LOH. Epithelial dysplasia is a sign of malignant risk, independent of lichenoid changes. Such results suggest that pathologists should search for dysplasia carefully in lesions that otherwise qualify as OLP and that caution should be used when discounting dysplasia as being merely a reactive condition in lichenoid lesions.

The application of optimal reactive power flow (ORPF) to the New Brunswick (NB) power system is presented. The potential benefits and the real-time implementation problems of ORPF are discussed. Some important issues of real-time implementation on ORPF, such as frequency of running, the number of activated control variables, and the order of adjustment of different controls, are discussed. The application of ORPF on the NB power network has shown two major benefits: (1) an improvement in the voltage profile and voltage stability, and (2) a saving in active power loss. The improvement in the voltage profile can cause fewer violations and a more stable system from the voltage point of view. A reduction in active power loss gained from ORPF can save a significant amount of money. The total ideal savings for the year 1997 predicted in the study was in excess of $900000, however, only 10 to 30% of this amount is realistically obtainable due to operational and other constraints. These savings can be gained simultaneously with the improvement of the voltage profiles.

The Scottish Intercollegiate Guidelines Network (SIGN) has developed guidelines for primary invasive cutaneous squamous cell carcinoma (cSCC). SIGN 140 Key questions included: (i) Which features, or combination of features, best identify high-risk cSCC? (ii) What are the most effective interventions for management of patients with cSCC? (iii) What is the optimum surgical margin and what histological clearance margins are acceptable? The evidence base was synthesised in accordance with SIGN methodology (SIGN 50), which complies with the internationally recognised AGREE criteria for guideline development based on a systematic review of the published literature. Databases searched included Medline, Embase, Cinahl and the Cochrane Library. Systematic searches were supplemented by material (mostly historic) identified by members of the development group. Initial searches identified 5712 hits, from which 492 abstracts were screened and 164 studies read in full and critically appraised before 91 were accepted as evidence on which to base recommendations. A systematic review and pooled analysis of observational studies was used as a basis for treatment recommendations. This process identified those clinical and histopathological features where there is robust evidence for association with poor outcomes (local recurrence, metastasis, disease-related death) allowing recognition of cSCC where referral to a regional skin cancer MDT meeting is appropriate. A minimum dataset highlighting these features would facilitate automatic referral and has been developed. Four key features; diameter, depth, differentiation and perineural invasion were identified onmultivariate analysis as associated with at least 2 of the 3 poor outcomes. Additional independent high-risk features identified were desmoplastic subtype, location on ear and immunosuppression. Recommendations and good practice points for surgical margins, Mohs Micrographic Surgery (MMS), radiotherapy and curettage have been developed, but the evidence base for surgical margins and for use of adjuvant radiotherapywas limited, with very little data relating surgical or histologicalmargins to outcomes. In retrospective analysis most incomplete excisions show persistence at the deep margin suggesting future studies should focus on deep as well as radial margins. Furthermore, the role

Fourteen cases of meningococcal disease (MD) occurred in August–September 1996 in western Sydney, Australia. Seven of the 10 young adults affected had a direct or indirect link with a local nightclub. Ten of 11 systemic meningococcal isolates had the phenotype C[ratio ]2a[ratio ]P1.5 and showed close genetic relationship by pulsed-fleld gel electrophoresis (PFGE). Organisms of this phenotype have not previously caused outbreaks in Australia, but have been associated with outbreaks and hyperendemic serogroup C MD in Europe, Canada, and the United States.This is the largest cluster of serogroup C MD reported in urban Australia, and the first involving a nightclub. The strain differentiation results were available rapidly enough to augment epidemiological investigations on a daily basis. Public health staff could thus establish links between cases quickly, follow the spread of new cases in the community, give accurate information to health officials and the press, and utilize existing knowledge about the characteristics of this phenotype to predict likely developments during the outbreak and afterwards. The strain differentiation data was also very helpful when the role of vaccination was considered, and existing guidelines on the management of outbreaks of MD could be used effectively for the first time in western Sydney.