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Estrogen receptor-alpha (ERα) regulates bone mass and is implicated in bone tissue's response to mechanical loading. The effects of ERα deletion in mice depend on sex, anatomical location, and the cellular stage at which ERα is removed. Few studies have investigated the effect of age on the role of ERα in skeletal maintenance and functional adaptation. We previously demonstrated that bone mass and adaptation to loading were altered in growing 10-week-old female and male mice lacking ERα in mature osteoblasts and osteocytes (pOC-ERαKO). Here our goal was to determine the effects of ERα and mechanical loading in skeletally-mature adult mice. We subjected 26-week-old skeletally-mature adult pOC-ERαKO and littermate control (LC) mice of both sexes to two weeks of in vivo cyclic tibial loading. ERα deletion in male mice did not alter bone mass or the response to loading. Adult female pOC-ERαKO mice had reduced cancellous and cortical bone mass and increased adaptation to high-magnitude mechanical loading compared to LC mice. Thus, ERα deletion from mature osteoblasts reduced the bone mass and increased the mechanoadaptation of adult female but not male mice. Additionally, compared to our previous work in young mice, adult female mice had greatly reduced mechanoadaptation and adult male mice retained most of their mechanoadaptation with age.

Objective Reduced subchondral bone mass and increased remodeling are associated with early stage OA. However, the direct effect of low subchondral bone mass on the risk and severity of OA development is unclear. We sought to determine the role of low bone mass resulting from a bone-specific loss of estrogen signaling in load-induced OA development using female osteoblast-specific estrogen receptor-alpha knockout (pOC-ERαKO) mice. Methods Osteoarthritis was induced by cyclic mechanical loading applied to the left tibia of 26-week-old female pOC-ERαKO and littermate control mice at peak loads of 6.5N, 7N, or 9N for 2 weeks. Cartilage damage and thickness, osteophyte development, and joint capsule fibrosis were assessed from histological sections. Subchondral bone morphology was analyzed by microCT. The correlation between OA severity and intrinsic bone parameters was determined. Results The loss of ERα in bone resulted in an osteopenic subchondral bone phenotype, but did not directly affect cartilage health. Following two weeks of cyclic tibial loading to induce OA pathology, pOC-ERαKO mice developed more severe cartilage damage, larger osteophytes, and greater joint capsule fibrosis compared to littermate controls. Intrinsic bone parameters negatively correlated with measures of OA severity in loaded limbs. Conclusions Subchondral bone osteopenia resulting from bone-specific loss of estrogen signaling was associated with increased severity of load-induced OA pathology, suggesting that reduced subchondral bone mass directly exacerbates load-induced OA development. Bone-specific changes associated with estrogen loss may contribute to the increased incidence of OA in post-menopausal women.

Mechanical loading is an anabolic stimulus that increases bone mass, and thus a promising method to counteract osteoporosis-related bone loss. The mechanism of this anabolism remains unclear, and needs to be established for both cortical and cancellous envelopes individually. We hypothesized that cortical and cancellous bone display different gene expression profiles at baseline and in response to mechanical loading. To test this hypothesis, the left tibiae of 10-week-old female C57Bl/6 mice were subjected to one session of axial tibial compression (9N, 1200 cycles, 4Hz triangle waveform) and euthanized 3 and 24 hours following loading. The right limb served as the contralateral control. We performed RNA-seq on marrow-free metaphyseal samples from the cortical shell and the cancellous core to determine differential gene expression at baseline (control limb) and in response to load. Differential expression was verified with qPCR. Cortical and cancellous bone exhibited distinctly different transcriptional profiles basally and in response to mechanical loading. More genes were differentially expressed with loading at 24 hours with more genes downregulated at 24 hours than at 3 hours in both tissues. Enhanced Wnt signaling dominated the response in cortical bone at 3 and 24 hours, but in cancellous bone only at 3 hours. In cancellous bone at 24 hours many muscle-related genes were downregulated. These findings reveal key differences between cortical and cancellous genetic regulation in response to mechanical loading. Future studies at different time points and multiple loading sessions will add to our knowledge of cortical and cancellous mechanotransduction with the potential to identify new targets for mouse genetic knockout studies and drugs to treat osteoporosis.

Background The American Board of Thoracic Surgery (ABTS) has noted a yearly decrease in the number of examination certificates being awarded, with only 93 certificates awarded in 2011. In 2003, the Accreditation Council for Graduate Medical Education required all programs to implement the 80-hour residency workweek. We hypothesized that this requirement has resulted in trainees being less capable of becoming successfully certified. Methods We examined the ABTS board scores, both written and oral, from 2000 to 2011. We divided the interval into 2 periods: 2000 to 2005, representing the 6-year, pre–80-hour workweek, and 2006 to 2011, the 6-year period post–80-hour workweek implementation. We analyzed whether a significant difference would be present in the pass rate before and after the 80-hour workweek for both the written and the oral boards. Results An inflection point of examination failures was found that started in 2006, correlating with the first examination year the 80-hour workweek would have affected. The written examination failure rates increased from 2006 to 2009 but have since decreased. The actual percentage failing the written component was less than the percentage failing the oral examinations in both periods. The oral examination failure rates have continued to increase at an alarming rate. Conclusions An increase has occurred in the failure of the ABTS board examinations that has been significantly greater after implementation of the 80-hour workweek. The failure rate for the written examination was not as significant as that for the oral examination. Because we are now training fewer, and perhaps less successful, cardiothoracic surgeons, it is our duty to develop strategies to improve and promote innovation in the methods of training.

Anastomotic leak is an important cause of morbidity and mortality after esophagectomy. Few studies have targeted risk factors for the development of leak after esophagectomy. The purpose of this study is to use The Society of Thoracic Surgeons Database to identify variables associated with leak after esophagectomy.The Society of Thoracic Surgeons Database was queried for patients treated with esophagectomy for esophageal cancer between 2001 and 2011. Univariate and multivariate analysis of variables associated with an increased risk anastomotic leak was performed.There were 7,595 esophagectomies, with 804 (10.6%) leaks. Thirty-day mortality and length of stay were higher for patients with anastomotic leak. Mortality in patients requiring surgical management was 11.6% (38 of 327) compared with 4.4% (20 of 458) in medically managed leaks (p0.001). The leak rate was higher in patients with cervical anastomosis compared with those with intrathoracic anastomoses, 12.3% versus 9.3%, respectively (p = 0.006). There was no difference in leak-associated mortality between the two approaches. Factors associated with leak on univariate analysis include obesity, heart failure, coronary disease, vascular disease, hypertension, steroids, diabetes, renal insufficiency, tobacco use, procedure duration greater than 5 hours, and type of procedure (p0.05). Multivariable regression analysis associated heart failure, hypertension, renal insufficiency, and type of procedure as risk factors for the development of leak (p0.05).Anastomotic leak after esophagectomy is an important cause of postoperative mortality and increased length of stay. We have identified important risk factors for the development of esophageal anastomotic leak after esophagectomy. Further studies aimed at risk reduction are warranted.

Background Discerning primary versus metastatic lung lesions is problematic. Comparative mutational profiling (CMP) involves genetic and point mutation analysis of lesions to facilitate this. We sought to review our experience in cases of two lung lesions or head and neck cancer and lung lesions to determine whether a significantly clinical problem existed, what standard processes were in place to address it, and whether a new diagnostic standard was required. Methods Between January 1, 2007, and October 31, 2008, CMP was used in 24 cases of two lung lesions or a head and neck cancer and lung lesion. Routine hematoxylin and eosin stain examination and immunohistochemistry were performed as appropriate. The CMP involved DNA sequencing for specific oncogene point mutations and a panel of allelic imbalance markers. Metastatic cancer required demonstration of concordant mutations affecting the same allele copy in different cancer deposits. Results The patient mean age was 62 years; there were 13 men and 11 women. The cases involved two lung lesions (n = 13) or a head and neck cancer and a lung lesion (n = 11). Standard pathology examination was unable to discriminate the lesions, and they were subsequently differentiated by CMP. Fifteen discordant CMP results were interpreted as independent primaries; 9 cases were concordant, consistent with metastatic disease. Conclusions Discerning primary versus metastatic disease when dealing with lung lesions is a clinically significant problem. Comparative mutational profiling was found to be useful and reliable to assess the relatedness of multiple cancer lesions when routine pathology assessment was unable to.

Background CT scan measures of emphysema and airway disease have been correlated with lung function in cohorts of subjects with a range of COPD severity. The contribution of CT scan-assessed airway disease to objective measures of lung function and respiratory symptoms such as dyspnea in severe emphysema is less clear. Methods Using data from 338 subjects in the National Emphysema Treatment Trial (NETT) Genetics Ancillary Study, densitometric measures of emphysema using a threshold of −950 Hounsfield units (%LAA-950) and airway wall phenotypes of the wall thickness (WT) and the square root of wall area (SRWA) of a 10-mm luminal perimeter airway were calculated for each subject. Linear regression analysis was performed for outcome variables FEV 1 and percent predicted value of FEV 1 with CT scan measures of emphysema and airway disease. Results In univariate analysis, there were significant negative correlations between %LAA-950 and both the WT ( r = −0.28, p = 0.0001) and SRWA ( r = −0.19, p = 0.0008). Airway wall thickness was weakly but significantly correlated with postbronchodilator FEV 1 % predicted (R = −0.12, p = 0.02). Multivariate analysis showed significant associations between either WT or SRWA (β = −5.2, p = 0.009; β = −2.6, p = 0.008, respectively) and %LAA-950 (β = −10.6, p = 0.03) with the postbronchodilator FEV 1 % predicted. Male subjects exhibited significantly thicker airway wall phenotypes (p = 0.007 for WT and p = 0.0006 for SRWA). Conclusions Airway disease and emphysema detected by CT scanning are inversely related in patients with severe COPD. Airway wall phenotypes were influenced by gender and associated with lung function in subjects with severe emphysema.

Tumor necrosis factor receptor-associated factor 6 (TRAF6) associates with the cytoplasmic domain of receptor activator of NF-κB (RANK) and is an essential component of the signaling complex mediating osteoclastogenesis. However, the osteoclastic activity of TRAF6 is blunted by its association with four and half LIM domain 2 (FHL2), which functions as an adaptor protein in the cytoplasm and transcriptional regulator in the nucleus. We find that TRAF6 also localizes in the nuclei of osteoclasts but not their bone marrow macrophage precursors and that osteoclast intranuclear abundance is specifically increased by RANK ligand (RANKL). TRAF6 nuclear localization requires FHL2 and is diminished in fhl2-/- osteoclasts. Suggesting transcriptional activity, TRAF6 interacts with the transcription factor RUNX1 in the osteoclast nucleus. FHL2 also associates with RUNX1 but does so only in the presence of TRAF6. Importantly, TRAF6 recognizes FHL2 and RUNX1 in osteoclast nuclei, and the three molecules form a DNA-binding complex that recognizes and transactivates the RUNX1 response element in the fhl2 promoter. Finally, TRAF6 and its proximal activator, RANKL, polyubiquitinate FHL2, prompting its proteasomal degradation. These observations suggest a feedback mechanism whereby TRAF6 negatively regulates osteoclast formation by intracytoplasmic sequestration of FHL2 to blunt RANK activation and as a component of a transcription complex promoting FHL2 expression.

SummaryMaintenance of bone mass and integrity requires a tight balance between resorption by osteoclasts and formation by osteoblasts. Exocytosis of functional proteins is a prerequisite for the activity of both cells. In the present study, we show that synaptotagmin VII, a calcium sensor protein that regulates exocytosis, is associated with lysosomes in osteoclasts and bone matrix protein-containing vesicles in osteoblasts. Absence of synaptotagmin VII inhibits cathepsin K secretion and formation of the ruffled border in osteoclasts and bone matrix protein deposition in osteoblasts, without affecting the differentiation of either cell. Reflecting these in vitro findings, synaptotagmin VII-deficient mice are osteopenic due to impaired bone resorption and formation. Therefore, synaptotagmin VII plays an important role in bone remodeling and homeostasis by modulating secretory pathways functionally important in osteoclasts and osteoblasts.

The molecular mechanisms by which binding of monocyte/macrophage colony-stimulating factor to its receptor c-Fms promotes replication in primary macrophages are incompletely understood, as all previous studies involved overexpression of receptor mutants in transformed cells not endogenously expressing the receptor. To address this issue we retrovirally expressed, in bone marrow-derived macrophages, a chimeric receptor containing a range of tyrosine to phenylalanine mutations in the c-Fms cytoplasmic tail. We measured incorporation of bromodeoxyuridine as a marker of proliferation and phosphorylation of ERKs, Akt, and the receptor itself. Our data indicate that tyrosine 559 is the major mediator of receptor activation and cell death, intracellular signaling, and cell proliferation and that the tyrosine residues at positions 697 and 807 play lesser roles in these events. Importantly, we find that activation of the ERK and Akt pathways is necessary but not sufficient for induction of macrophage proliferation. Using specific small molecule inhibitors we find that a combination of the Src family kinase, phosphatidylinositol 3-kinase/Akt, phospholipase C, and ERK pathways mediates macrophage proliferation in response to M-CSF.

Studies have shown a potential role for humoral rejection in the evolution of lung graft dysfunction, apparently based on antibodies without human leukocyte antigen specificity. The correlation between extent of C4d deposition with clinical status further illustrates the importance of humoral immunity. Our study examines the potential value of C3d as a further diagnostic adjunct. C3d deposition was examined in lung allograft specimens using frozen tissue indirect direct immunofluorescence (IIF) and avidin biotin immunohistochemical applied to paraffin embedded tissue. Intermediate/extensive amounts of C3d using IIF and immunohistochemical (IH) methodologies correlated with chronic graft dysfunction; IIF C3d deposition was associated with septal and bronchial wall fibrosis (p < 0.0001). Weak/absent amounts of IIF and IH C3d correlated with clinical stability (p < 0.0001). Higher levels of C3d by IH were more sensitive than by IIF as a bronchiolitis obliterans syndrome determinant. C3d and C4d deposition using immunofluorescence and IH were correlated (p < 0.00001). C3d deposition appears prognostically significant. Higher tissue expression of C3d mark chronic graft dysfunction/persistent graft failure following transplantation. The close correlation between C3d and C4d lends credence to the role of humoral allograft rejection as a pulmonary graft dysfunction contributing factor. C3d by IH manifests higher sensitivity but similar specificity compared to C3d by IIF.

Primary intrapulmonary nodular amyloidosis is a rare form of immunoglobulin associated amyloidosis, also falling under the alternative appellation of amyloidoma. Although amyloidomas in other organ sites may be reflective of a localized or more generalized plasma cell dyscrasia, in the context of its presentation in the lung the presumptive basis has long been held to be one of chronic inflammation. We encountered 2 patients with nodular amyloidosis in whom the pathologic examination disclosed the basis to be one of a light chain restricted clonal lymphocytic plasma cell infiltrate, although without morphologic features of coexisting pulmonary lymphoma. These 2 patients serve to underscore the potential categorization of some cases of pulmonary nodular amyloidosis as a form of low grade B cell lymphoproliferative disease.

Background Histoplasma capsulatum is a dimorphic fungus with both hyphal and microconidial elements and is endemic in certain areas of the central United States. Most clinically recognized syndromes are self-limited, presenting in more than 80% of cases as an influenza-like acute pulmonary illness. We encountered four patients with an endobronchial presentation of histoplasmosis masquerading as a bronchogenic malignancy. Methods The patient files from one of the authors (PR) uncovered four patients diagnosed with, and treated for, endobronchial histoplasmosis; all cases were referred for a presumptive diagnosis of endobronchial neoplasia. Results The case study group comprised three women, ages 52, 68, and 81 years, respectively, and one man age 29 years, all presenting with recurrent hemoptysis. An obstructing adherent lesion was apparent on bronchoscopy in three, involving the right upper lobe bronchus in two, and right mainstem bronchus in one. In one patient, blood was observed in the bronchus intermedius; there was no gross lesion. Recurrent hemoptysis necessitated a right upper sleeve lobectomy in one, a right upper lobe lobectomy and right lower wedge resection in one, a right middle lobectomy in one, and wedge resection of the right upper lobe in one. Conclusions Endobronchial histoplasmosis may mimic primary bronchogenic carcinoma, presenting as a fixed obstructing endobronchial lesion associated with hemoptysis; the latter potentially necessitating surgical intervention. Endobronchial histoplasmosis should be considered in the differential diagnosis of obstructing endobronchial lesions associated with hemoptysis, especially when prior and(or) concurrent biopsies fail to disclose malignancy.

Endobronchial presentation of Hodgkin lymphoma is exceedingly rare and can be confused clinically with pulmonary small cell carcinoma. Because of the dramatic implications in treatment and prognosis, endobronchial Hodgkin lymphoma, despite its uncommon occurrence, should be considered in the differential of small cell carcinoma and necrotizing vasculitides with pulmonary involvement, especially in a relatively young patient with cough, hemoptysis, atelectasis, and hilar or mediastinal lymphadenopathy. The use of photodynamic laser therapy or stent placement for palliative treatment of life-threatening airway obstruction may be required prior to or as an initial adjunct to tumor-specific therapy.

Animal studies have shown that blockade of complement may reduce the severity of and/or prevent the development of bronchiolitis obliterans syndrome (BOS), suggesting a role for complement activation. We explored the hypothesis that humoral immunity plays a role in the evolution of BOS. Thirteen unilateral lung transplant patients with BOS defined the patient population. Fresh frozen tissue was analyzed for deposition of C1q, C4d, C5b-9 and immunoglobulin (IgG, IgM, IgA). An indirect immunofluorescent assay was also conducted with patient serum against cytospins of the pulmonary endothelium. In each case the biopsies showed a microvascular injury syndrome involving the bronchial wall characterized by one or more of hemorrhage, fibrin deposition, and endothelial cell necrosis. Other features included bronchial epithelial and chondrocyte necrosis. The end-stage lesion was a thinned bronchial epithelial lining mural fibrosis. Immunofluorescent analysis showed deposition of C1q, C3, C4d, C5b-9, and immunoglobulin in the bronchial epithelium, chondrocytes, basement membrane zone of the bronchial epithelium, and bronchial wall microvasculature. The indirect antiendothelial cell antibody assay was positive in all tested. Humoral immunity may play a role in the pathogenesis of BOS; the antigenic targets include the bronchial wall microvasculature, bronchial epithelium, and chondrocytes.

Primary esophageal schwannomas are uncommon. We describe a case of a large asymptomatic primary esophageal schwannoma in a 65-year-old patient. Computed tomography and positron emission tomography revealed an (18)F-fluorodeoxyglucose-avid 11-cm mass arising from the esophagus. A preoperative diagnosis was made via endoscopic ultrasound. The patient underwent a three-field esophagogastrectomy with cervical esophagogastric anastomosis. He remains well and free of recurrence 10 months after treatment.

Reactive lymphoid hyperplasia is a common pathological finding from lung biopsy. It is frequently the original pathologic interpretation from FNA or core biopsies of lesions that are ultimately determined to be non-neoplastic pulmonary lymphoid disorders. One of these disorders is Castleman's Disease. We present a case of a 35-year-old male who underwent multiple procedures to diagnose a posterior mediastinal mass. This was eventually diagnosed as Castleman's Disease. We review the pathology of reactive lymph node hyperplasia and the spectrum of non-neoplastic pulmonary lymphoid disorders. We also review the Unicentric hyaline vascular variant form of Castleman's disease.

Murine osteoclast precursors and osteoblasts express the integrin alpha(v)beta(5), the appearance of which on the cell surface is controlled by the beta(5), and not the alpha(v), subunit. Here, we show that a 173-base pair proximal region of the beta(5) promoter mediates beta(5) basal transcription in macrophage (osteoclast precursor)-like and osteoblast-like cells. DNase I footprinting reveal four regions (FP1-FP4) within the 173-base pair region, protected by macrophage nuclear extracts. In contrast, osteoblast nuclear extracts protect only FP1, FP2, and FP3. FP1, FP2, and FP3 bind Sp1 and Sp3 from both macrophage and osteoblast nuclear extracts. FP4 does not bind osteoblast proteins but binds PU.1 from macrophages. Transfection studies show that FP1 and FP2 Sp1/Sp3 sites act as enhancers in both MC3T3-E1 (osteoblast-like) and J774 (macrophage-like) cell lines, whereas the FP3 Sp1/Sp3 site serves as a silencer. Mutation of the FP2 Sp1/Sp3 site totally abolishes promoter activity in J774 cells, with only partial reduction in MC3T3-E1 cells. Finally, we demonstrate that PU.1 acts as a beta(5) silencer in J774 cells but plays no role in MC3T3-E1 cells. Thus, three Sp1/Sp3 sites regulate beta(5) gene expression in macrophages and osteoblast-like cells, with each element exhibiting cell-type and/or activation-suppression specificity.

Hyperammonemia after lung transplantation is a rare complication of unknown etiology. Its management is largely supportive and outcomes have been variable. More disconcerting is its immunosuppressive management because the precipitating factors leading to this potentially lethal entity are unknown, but are suspected to be drug-related. We describe the successful management of a lung transplant recipient with severe hyperammonemia.

We previously noted that the initial receptor by which murine osteoclast precursors bind matrix is the integrin alphav beta5 and that granulocyte-macrophage colony-stimulating factor (GM-CSF) decreases expression of this heterodimer by suppressing transcription of the beta5 gene. We herein report cloning of the beta5 integrin gene promoter and identification of a GM-CSF-responsive sequence. A 13-kilobase (kb) genomic fragment containing part of the beta5 gene was isolated by screening a mouse genomic library with a probe derived from the most 5'-end of a murine beta5 cDNA. A combination of primer extension and S1 nuclease studies identifies two transcriptional start sites, with the major one designated +1. A 1-kb subclone containing sequence -875 to + 110 is transcriptionally active in a murine myeloid cell line. This 1-kb fragment contains consensus binding sequences for basal (Sp1), lineage-specific (PU.1), and regulatable (signal transducer and activator of transcription) transcription factors. Reflecting our earlier findings, promoter activity is repressed in transfected myeloid cells treated with GM-CSF. Using deletion mutants, we localized a 109-base pair (bp) promoter region responsible for GM-CSF-inhibited beta5 transcription. We further identified a 19-bp sequence within the 109-bp region that binds GM-CSF-induced nuclear proteins by gel shift/competition assays. Mutation of the 19-bp sequence not only ablates its capacity to bind nuclear proteins from GM-CSF-treated cells, in vitro, but the same mutation, when introduced in the 1-kb promoter, abolishes its ability to respond to GM-CSF treatment. Northern analysis demonstrates that cycloheximide treatment abrogates the capacity of GM-CSF to decrease beta5 mRNA levels. In summary, we have identified a 19-bp cis-element mediating GM-CSF-induced down-regulation of beta5 by a mechanism requiring protein synthesis.

Tumor necrosis factor-α (TNF) exerts its transcriptional effects via activation of nuclear transcription factor-κ B (NF-κB). NF-κB is sequestered in the cytosol by IκBα and, in most cells, released upon serine phosphorylation of this inhibitory protein which then undergoes rapid, ubiquitin-dependent degradation. In contrast, we find TNF induction of NF-κB in murine bone marrow macrophages (BMMs), is mediated, by c-Src, in a cell, and cytokine specific manner. The non-receptor tyrosine kinase is rapidly mobilized and activated upon TNF exposure. Within the same time frame, TNF induced c-Src associates with IκBα in a long lived complex. The proto-oncogene, when associated with IκBα phosphorylates the inhibitory protein on tyrosine 42. Consistent with the pivotal role played by c-Src in TNF-induced IκBα tyrosine phosphorylation, NF-κB activation, by the cytokine, is markedly delayed and reduced in c-src−/− BMMs. Underscoring the physiological significance of c-Src activation of NF-κB, TNF induction of IL-6, which is an NF-κB mediated event, is substantially diminished in c-src−/− BMMs.

We have identified a novel steroid hormone response element in the avian beta3 integrin promoter. This sequence, comprising three hexameric direct repeat half-sites separated by nine and three nucleotides binds vitamin D receptor (VDR)-retinoid X receptor (RXR) and retinoic acid receptor (RAR)-RXR heterodimers. VDR-RXR binds direct repeats separated by three base pairs, and RAR-RXR recognizes half-sites separated by nine bases, whereas the central half-site interacts with both heterodimers. Retinoic acid and 1, 25-dihydroxyvitamin D3 activate both a genomic fragment including the transcriptional start site and an oligonucleotide containing the three repeats, linked to a heterologous promoter. Co-addition of the steroids produces neither synergy nor an additive effect; rather the result equals that for retinoic acid alone. Scatchard analysis demonstrates that RAR-RXR has greater affinity than VDR-RXR for the composite element. Based on these findings we propose a model in which there is specific, polarity-defined binding of VDR-RXR and RAR-RXR to three half-sites, which form two overlapping steroid response elements, with the central half-site common to both. Our results identify a novel mechanism by which one steroid hormone can modulate the activity of a second, by competing for a shared half-site in a composite response element.

Lung cancer remains a major public health problem because it is the leading cause of cancer death in men and women. The traditional surgical role for lung cancer has been for patients with limited disease; that approach resulted in an overall five year survival of 18–20%. During the past decade, increasing attention has been paid to expanding the group of patients for whom surgical resection could be accomplished. The net effect of aggressive surgical intervention, addition of multimodality therapy, and improved understanding of perioperative and postoperative management has been improved survival in stages I, II, and III. At this time a non-small cell cancer of the lung should be surgically staged and based on stage, therapy should be designed. Multimodality therapy is an important adjunct to stage IIIA and stage IIIB patients. With a multidisciplinary approach to lung cancer, patients in stages I, II, IIIA, and IIIB may all be candidates for resection and cure. Patients in stage IV are candidates for palliative intervention although no long-term impact on survival has been achieved. Early detection, prompt evaluation, surgical staging, and multidisciplinary therapy are the current standards by which the overall outcome of patients with non-small cell cancer of the lung will be improved.

Osteoclastic bone resorption is dependent upon cell-matrix recognition. This process is mediated by the integrin αvβ3 whose expression is enhanced, in avian osteoclast precursors, by bone-seeking steroids. The purpose of this study was to determine if bone-modulating cytokines impact on αvβ3 expression by mouse marrow macrophages (BMMs), known to differentiate into osteoclasts. Of the cytokines tested. Interleukin-4 (IL-4) is most effective in increasing β3 mRNA levels by a mechanism involving transactivation of the β3 gene. Moreover, IL-4 augmented β3 mRNA is mirrored by plasma membrane appearance of αvβ3. As IL-4 induces β3 and not αv mRNA, the β3 chain appears to regulate surface expression of the heterodimer. The functional significance of IL-4-induced αvβ3 is underscored by the fact that, while attachment to fibronectin is unaltered, treatment of BMMs with the cytokine enhances αvβ3-mediated binding to vitronectin 5-fold. Expression of this heterodimer by BMMs driven along a non-osteoclastic lineage suggests αvβ3 may play a role in the inflammatory response of macrophages.

Endobronchial presentation of Hodgkin lymphoma is rare and can be confused clinically and radiographically with pulmonary small cell carcinoma. We present a case of an obstructive endobronchial presentation of nodular sclerosing classic Hodgkin lymphoma, initially misdiagnosed as small cell carcinoma, with endobronchial vasculitis and associated hemoptysis. Photodynamic therapy relieved the obstruction before induction of tumor-specific therapy. This case demonstrates the successful use of photodynamic therapy in obstructive endobronchial Hodgkin lymphoma.

During the past decade, we have increased greatly our understanding of the pathogenesis of postmenopausal osteoporosis, a major cause of morbidity and mortality. We know now that estrogen regulates the expression of cytokines that target cell types involved in modulating bone turnover. Although early work demonstrated that tumor necrosis factor α plays an important role in regulating bone mass, recent studies also implicate the lymphopoietic molecule interleukin 7. This protein is unique in that it suppresses the bone-forming osteoblast, whilst stimulating formation and function of the osteoclast, the exclusive resorptive cell. Furthermore, the latest findings confirm and expand the concept that T cells are key mediators of bone loss following gonadal failure.

The appearance of the vitamin D-dependent calcium-binding protein (calbindin-D28K) in embryonic chick tissues was determined using a sensitive immunohistochemical assay to elucidate the possible role of calbindin-D28K during embryogenesis. This protein first occurs in renal, cerebellar and intestinal cells during the final stages of maturation and/or differentiation. It is during this period of their development that these tissues become functional. On the basis of our observations, we suggest that the onset of cell function may be coupled with the appearance of calbindin-D28K.

1. 1. Immunohistochemical examination of sections of adult bat (Rousettus aegyptiacus) kidney failed to show the expression of calbindin-D28K in any tubules. 2. 2. Calbindin-D9K was found to occur in the distal convoluted tubules. 3. 3. Purification of the putative calbindin-D9K protein revealed its migration in non-denaturing polyacrylamide gel electrophoresis (PAGE) to be Ca-sensitive. 4. 4. On 17% SDS-PAGE gels the protein had an apparent M r of less than 12,000. 5. 5. The absence of calbindin-D28K from the kidney is unusual, but until roles are assigned to calbindins the significance of this finding will remain unclear.

s / Photodiagnosis and Photodynamic Therapy 12 (2015) 325–375 367 Photodynamic field control of malaria vector, schistosomiasis and agricultural pests Mahmoud H. Abdel Kader The German University in Cairo, Egypt In this work, we present the successful field implementation of using the photodynamicmodality to control vector-borne diseases, suchasMalaria, Filaria,Dengue feverandSchistosomiasis, aswell as agro-insects which cause considerable damage to agro-economics. Field trials were performed in infested epidemic swamps in Uganda, Ethiopia and Sudan using chlorophyll derivatives as a photosensitizer which was approved by the FDA as food additives. It was added to the infested swamps to be uptaken by the mosquito larvae. The accumulated chlorophyll derivatives inside the larvae body induce, upon sunlight exposure, oxidative stress, which interrupts the life cycle of theparasite and results in organismdeath. The results revealed85–100%mortality of larvaepopulationat different concentrations of chlorophyll derivatives. The formulated photosensitizerusedachieved target selectivitywhereall otherbiological beneficiary organisms (which were present in the same treated swamps) were not affected. Using the same technique for the snail vector control (intermediate host of Schistosomiasis (Biomphalaria alexandrina), mortality percentage obtained ranges from 80% to 90%. In case of agro-insects, two different types of nano-materials (Graphene-oxide and Silver nanoparticles were conjugated with Magnesium and Copper Chlorophyllin derivatives which are used as photo-insecticide against white-fly, Cotton Worm and fruit-fly. Field results revealed that all nanocomposites achieved high reduction percentages in insects’ population ranging from 75% to 96.5%. The results reveals that this innovative modality against vector borne diseases as well as agro-pests combines both effectiveness and efficiency with the highest levels. http://dx.doi.org/10.1016/j.pdpdt.2015.07.163 Physiology properties as predictors of PDT response: Where we stand and where we are going

Cervical cancer is the third most commonly diagnosed cancer and the fourth leading cause of cancer death in womenworldwide. The highest incidence rates are in Africa, as well as South-Central Asia and South America. According to the National Institute of Cancer (INCA) at the last year, it was estimated 15,590 new cases of cervical cancer in Brazil. About 90% of cervical cancers originate in the evolution of cervical intraepithelial neoplasia (CIN) which is classified on the basis of cyto-histological characteristics (low and high grade lesions). The equipment (CerCa 150 System, MMOptics Ltda., Trubios LLC, Brazil-USA) and the medication (20% MAL cream, PDTPharma, Brazil) are produced in Brazil with relative low cost. The Photodynamic Therapy is a promising procedure and it is even known as safe to normal tissue. The device includes two components specifically adapted to detect and treat abnormal cervical tissue, enabling proper lighting of the cervix. It is portable and does not require special infrastructure or installation. More than 60 patients between the ages of 14-57 years and with CIN 1 were treated and presented 100% of cure. Based on these results a larger randomized study with patients with CIN 2, placebo and follow-up only is ongoing.

Growing consumer concern regarding the perceived inadequate privacy and security of personal information in light of 2005 data breaches can depress consumer spending, but given the tremendous personal and economic benefits that flow from private markets, government officials must resist the urge to 'do something' to quickly satisfy political needs with ineffective or even harmful legislation that can also stifle economic growth. An effective role for the federal government is to work closely with the private sector to educate businesses and consumers and to create a legal framework to override conflicting laws. In addition, the private sector should implement aggressive self regulation and modify management processes to make respect for privacy and protection of personal information a daily part of the corporate culture and way of doing business. Consumers should take responsibility for their role in securing the internet by using available security tools, safe practices and information that can substantially reduce vulnerabilities. In addition to these observations, three key concepts that are vital to any future security solutions involve a commitment to the continued legitimate flow of information that drives today's commerce, an enhanced respect for individual privacy and a commitment to protect sensitive information, and effective means to minimize vulnerabilities to this vital part of our critical national infrastructure and security.

Neither side of the intellectual property debate can define what is best for culture or society. Instead, an artist's control over her work, enforced by intellectual property rights, allows culture to spread and thrive. It isn't necessary for self-appointed defenders of culture to decide what is in the best interests of the artist. Furthermore, digital delivery of content should not dilute the rights of artists. Citing the history of intellectual property law, and the subsequent flourish of culture in the U.S., illustrates that giving artists a limited monopoly on their work encourages future creation. In the U.S., our views and laws on intellectual property stem from philosophers of physical property. William Blackstone, heavily influenced by John Locke's idea of property as an inalienable right, first gave us the concept of the Blackstonian bundle, an idea vital to intellectual property. The Blackstonian bundle compares individual property rights to a bundle of sticks, which can be leased sold, traded or given away. Culture has thrived even as artists alternately extended and withheld sticks from their Blackstonian bundle. The copyright debate has gathered momentum with the widespread adoption of the Internet to distribute content. However for respected scholars to argue that depriving a creator of potential revenue for her work is harmless to the artist shows a true disrespect to all artists, and a dangerous compulsion to put the desires of the masses ahead of the rights of the individual. Instead, the Internet has created an increase in niche markets because of the falling cost of distribution and production of online content. Therefore, creators of specialized content now have the ability to profit from their work.

expression of SMAD4 characterized patient derived xenografts (PDX), and identify profiles that may predict response to radiation. Materials/Methods:With IACUC approval, 20 PDAC PDXs characterized for KRAS, p53 and SMAD4 were implanted into bilateral flanks of mice and irradiated. Left flank tumors received 5 Gy/day for 2 days using the right flank as a control (n Z 5 per group). Tumor dimensions were obtained twice weekly (volumeZ (LxW2)/2); tumor growth inhibition (TGI) was calculated (treated tumor volume/control tumor volume). Gene expression data (Affymetrix GeneChip) was preprocessed with quantile normalization and log2 transformation. Methylation microarrays (Illumina 450k) were processed with quantile normalization, beta-value scale, and M-value. Key pathways of differentially expressed genes were investigated with KEGG pathway enrichment analysis. Results: Comparison of SMAD4 mutant (n Z 10) and intact (n Z 10) tumors revealed 621 and 602 differentially expressed genes following gene expression and methylation array analysis, respectively. Prominent KEGG pathways included endocytosis, melanogenesis, tight junction, ascorbate and aldarate metabolism, steroid hormone biosynthesis, and androgen and estrogen metabolism. Three genes with the strongest inverse correlation between levels of methylation and mRNA expression included ACOX2 (r Z -.56, p Z .01), CTAGE5 (r Z -.7, p Z .0004), and LY75 (r Z .61, p Z .004). Gene expression and methylation analysis as a function of relative TGI revealed 301 and 447 differentially expressed genes, respectively, and these were enriched in the pathways in cancer, melanoma, and aldosterone-regulated sodium reabsorption pathways. Irradiated tumors revealed a varying range of TGI (-58% to +72%). Using univariate linear regression to correlate TGI with gene expression and methylation revealed 5 target genes (C6orf125, CFD, DNAI1, GLI3, and TRIM24) with strong inverse correlation (r Z .53-.61, p