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Bayluscide, a chemical lampricide, is used by government agencies in the Great Lakes basin to assess Sea Lamprey (Petromyzon marinus) as part of bi-national efforts to control the species. The use of granular Bayluscide has been highly successful in identifying and suppressing Sea Lamprey populations. However, the ecological risk of Bayluscide for fishes and mussels listed under Canada's Species at Risk Act and other species of conservation concern is not well understood. This document: 1) identifies potential pathways (direct, indirect) and mechanisms (physiological, non-physiological) by which the application of granular Bayluscide may influence fishes and mussels of conservation concern; 2) evaluates the relative ecological risk of applications based on four metrics (spatial distribution, application intensity, habitat associations that predispose species to exposure, toxicity); 3) identifies mitigation measures that may reduce potential impacts to non-target species; and, 4) identifies uncertainties needed to refine risk estimates. Spatial analyses indicated that between 2011 and 2017, Bayluscide applications occurred within the distribution of 21 fish and 15 mussel species of conservation concern including areas identified as critical habitat for 16 species (6 fishes, 10 mussels). For fishes, relative risk was greatest for native lampreys (Silver Lamprey [Ichthyomyzon unicuspis] and Northern Brook Lamprey [Ichthyomyzon fossor]) followed by Lake Sturgeon (Acipenser fulvescens) and Northern Madtom (Noturus stigmosus). Native lampreys exhibited high relative risk given habitat preferences and the toxicity of Bayluscide to those species. Northern Madtom ranked highly due to exposure (spatial and temporal patterns of application) and toxicity. Lake Sturgeon ranked highly due to exposure, though results were not directly comparable with other species due to assessment methods. For mussels, relative risk was greatest for Salamander Mussel (Simpsonaias ambigua), Threehorn Wartyback (Obliquaria reflexa), and Hickorynut (Obovaria olivaria). Salamander Mussel and Threehorn Wartyback ranked highly due to exposure and toxicity. A high risk ranking for Hickorynut occurred due to exposure and habitat preference. Mitigation measures to reduce the ecological consequences of Bayluscide applications are numerous and include modifying the frequency and timing of treatments, decreasing the size of application sites, and avoiding areas near critical habitat. However, the effect of potential mitigation measures should be rigorously tested to ensure that desired outcomes for species of conservation concern are realized while avoiding unintended consequences. Lastly, as reduced control effectiveness of Sea Lamprey would have undesirable effects on species of conservation concern susceptible to parasitism by Sea Lamprey (e.g., Lake Sturgeon), optimization measures may be warranted to account for such trade-offs. [ABSTRACT FROM AUTHOR]

Pygmy Whitefish (Prosopium coulterii) was first captured in Lake Superior in 1952. In November 2016, COSEWIC assessed Pygmy Whitefish, Great Lakes-Upper St. Lawrence populations (DU5) as Threatened. The reason given for this designation was that "this small-bodied freshwater fish has experienced dramatic declines in abundance over the last several decades, with an overall estimated decline of 48% since 2000. The continued presence of invasive fishes and recovery of native predatory fishes may threaten or limit recovery, respectively" (COSEWIC 2016). The Recovery Potential Assessment (RPA) provides background information and scientific advice needed to fulfill various requirements of the federal Species at Risk Act. This research document provides the current state of knowledge of the species including its biology, distribution, population trends, habitat requirements, and threats, which will be used to inform recovery plans. A population status assessment indicated that the density of Pygmy Whitefish in Lake Superior has declined since 2013. A threat assessment indicated that the greatest threats to Pygmy Whitefish within DU5 are climate change, invasive species, and pollution; however, the impact of these threats is currently unknown. Predation from top predators such as Lake Trout (Salvelinus namaycush) and Burbot (Lota lota) may be limiting population growth. [ABSTRACT FROM AUTHOR]

Highlights • Alisertib combined with concurrent fractionated stereotactic radiation therapy (FSRT) is well tolerated for patients with recurrent high grade gliomas (HGG). • Clinical outcomes are similar to historical results from alternative treatment options for recurrent HGG. • Further exploration combining FSRT with novel systemic therapy agents may provide promising pathways for managing recurrent HGG. Abstract Background and purpose We conducted a phase I trial of alisertib, an oral aurora kinase inhibitor, with fractionated stereotactic re-irradiation therapy (FSRT) for patients with recurrent high grade glioma (HGG). Materials and methods Adult patients with recurrent HGG were enrolled from Feb 2015 to Feb 2017. Patients were treated with concurrent FSRT and alisertib followed by maintenance alisertib. Concurrent alisertib dose was escalated from 20 mg to 50 mg twice daily (BID). Results 17 patients were enrolled. Median follow-up was 11 months. Median FSRT dose was 35 Gy. There were 6, 6, 3, and 2 patients enrolled in 20 mg, 30 mg, 40 mg, and 50 mg cohort, respectively. Only one DLT was observed. One patient in the 20 mg cohort had severe headache (Grade 3) resolved with steroids. There was no non-hematological grade 3 or higher toxicity. There were two Grade 4 late toxicities (one with grade 4 neutropenia and leukopenia, one with pulmonary embolism). One patient developed radiation necrosis (Grade 3). Sixteen patients finished concurrent treatment and received maintenance therapy (median cycles was 3, range 1–9). OS for all cohorts at 6 months was 88.2% with median survival time of 11.1 months. PFS at 6 months was 35.3% with median time to progression of 4.9 months. The trial stopped early due to closure of alisertib program with only 2 of 3 planned patients enrolled in the 50 mg cohort. Conclusion Re-irradiation with FSRT combined with alisertib is safe and well tolerated for HGG with doses up to 40 mg BID. Although no DLT observed in the 50 mg cohort, this cohort was not fully enrolled and MTD was not reached. Clinical outcomes appear comparable to historical results. (NCT02186509) [ABSTRACT FROM AUTHOR]

Purpose: To determine the impact on overall survival with different salvage therapies, including no treatment, reirradiation, systemic therapy, or radiation and systemic therapy, in participants of a phase 3 clinical trial evaluating dose-dense versus standard-dose temozolomide for patients with newly diagnosed glioblastoma.Methods and Materials: This analysis of patients from Trial RTOG 0525 investigated the effect of reirradiation or systemic treatment after tumor progression. Survival from first progression was compared between patients receiving no therapy, systemic therapy alone, radiation alone, and both modalities. The Cox proportional hazards model was used to compare the mortality hazard, controlling for potential confounders.Results: The analysis included 637 patients who progressed and had information on their management, excluding those who died less than half a month after progression. A total of 267 patients (42%) received neither reirradiation nor systemic treatment at progression, 24 (4%) received radiation alone, 282 (44%) received systemic treatment only, and 64 (10%) received both radiation and systemic therapy. Patients who received no treatment had a median survival of 4.8 months, lower than with radiation treatment alone (8.2 months), systemic therapy alone (10.6 months), and both radiation and systemic therapy (12.2 months). In survival models controlling for potential confounders, those who received radiation alone had modestly better survival (hazard ratio HR 0.74, 95% confidence interval [CI] 0.43-1.28), whereas those who underwent systemic therapy either without (HR 0.42, 95% CI 0.34-0.53) or with radiation therapy (HR 0.44, 95% CI 0.30-0.63) had better survival. There was no significant survival difference between patients who received radiation only and those who received systemic therapy (either with radiation or alone).Conclusions: Patients who received no salvage treatment had poorer survival than those who received radiation, chemotherapy, or the combination. However, patient selection for no treatment likely reflects poorer expected prognosis. There was no significant survival difference among those receiving radiation therapy, systemic therapy, or both. Ongoing clinical trials will help define the role of reirradiation after glioblastoma progression. [ABSTRACT FROM AUTHOR]

By comparing the structures of Bax and Bak megapores, Cosentino et al. (2022) reveal new insights suggesting the two pro-apoptotic proteins co-assemble into structures that release DNA from mitochondria and thereby trigger inflammation. [ABSTRACT FROM AUTHOR]

Background: Skull base meningiomas commonly present with cranial neuropathies. Fractionated stereotactic radiation therapy (FSRT) has been used to treat these tumors with excellent local control, but rates of improvement in cranial neuropathies have not been well defined. We review the experience at Thomas Jefferson University using FSRT in the management of these patients with a focus on symptom outcomes. Methods: We identified 225 cases of skull base meningiomas treated with FSRT at Thomas Jefferson University from 1994 through 2009. The target volume was the enhancing tumor, treated to a standard prescription dose of 54 Gy. Symptoms at the time of RT were classified based on the cranial nerve affected. Logistic regression was performed to determine predictors of symptom improvement after FSRT. Results: The median follow-up time was 4.4 years. In 92% of cases, patients were symptomatic at the time of RT; the most common were impaired visual field/acuity (58%) or extraocular movements (34%). After FSRT, durable improvement of at least one symptom occurred in 57% of cases, including 40% of visual acuity/visual field deficits, and 40% of diplopia/ptosis deficits. Of all symptomatic patients, 27% experienced improvement of at least one symptom within 2 months of the end of RT. Conclusions: FSRT is very effective in achieving improvement of cranial neuropathies from skull base meningiomas, particularly visual symptoms. Over half of treated patients experience a durable improvement of at least one symptom, frequently within 2 months from the end of RT [ABSTRACT FROM AUTHOR]

Significant involvement of students in the development and implementation of college alcohol prevention strategies is largely untested, despite recommendations by the National Institute of Alcohol Abuse and Alcoholism and others. Objective: The purpose of the Pragmatics Project was to test a student engagement model for developing and implementing alcohol intervention strategies. Participants: The Pragmatics Project involved 89 undergraduate students on a large Midwestern university campus in the design and implementation of projects focused on reducing harm associated with high-risk drinking and off-campus parties. Results: The engagement model used an innovative course piloted in the Human Development and Family Science department. The course successfully involved both students and the community in addressing local alcohol issues. Conclusions: The course design described would fit well into a Master of Public Health, Community Psychology, Health Psychology, or interdisciplinary curricula as well as the service learning model, and it is applicable in addressing other health risk behaviors. [ABSTRACT FROM AUTHOR]

Purpose: To describe our initial experience of fractionated stereotactic radiotherapy dose reduction comparing two dose cohorts with examination of tumor control rates and serviceable hearing preservation rates. Methods and Materials: After institutional review board approval, we initiated a retrospective chart review to study the hearing outcomes and tumor control rates. All data were entered into a JMP, version 7.01, statistical spreadsheet for analysis. Results: A total of 89 patients with serviceable hearing had complete serial audiometric data available for analysis. The higher dose cohort included 43 patients treated to 50.4 Gy with a median follow-up (latest audiogram) of 53 weeks and the lower dose cohort included 46 patients treated to 46.8 Gy with a median follow-up of 65 weeks. The tumor control rate was 100% in both cohorts, and the pure tone average was significantly improved in the low-dose cohort (33 dB vs. 40 dB, p = 0.023, chi-square). When the patient data were analyzed at comparable follow-up points, the actuarial hearing preservation rate was significantly longer for the low-dose cohort than for the high-dose cohort (165 weeks vs. 79 weeks, p = .0318, log–rank). Multivariate analysis revealed the dose cohort (p = 0.0282) and pretreatment Gardner-Robertson class (p = 0.0215) to be highly significant variables affecting the hearing outcome. Conclusion: A lower total dose at 46.8 Gy was associated with a 100% local control tumor rate and a greater hearing preservation rate. An additional dose reduction is justified to achieve the optimal dose that will yield the greatest hearing preservation rate without compromising tumor control for these patients. [Copyright &y& Elsevier]

Abstract: Background: Stereotactic radiosurgery and fractionated stereotactic radiotherapy have become widespread techniques applied to the treatment of a variety of intracranial lesions. Rapid evolution of new technologies has now enabled clinicians to treat tumors outside the cranium and down the spinal axis. This review compares 3 commercially available systems in widespread use throughout the world. Methods: Literature review and interviews with practitioners in the United States were performed to establish data for a comparative analysis of the Gamma Knife (Elekta, Sweden), Novalis (BrainLabs, Germany), and CyberKnife systems (Accuray, Sunnyvale, CA). Cost analyses were deliberately excluded because of the need for detailed cost-benefit analysis beyond the scope of the review. Results: An unbiased comparative analysis was not possible because of the lack of objective data from a standard metric for these systems. Despite this shortcoming, disparate features of each system were compared and contrasted. Conclusion: A careful assessment of each system, including its operational features, capabilities, and yearly capacity must be weighed against the composition of the radiosurgery team, the case mix of the practice, and the objectives of the clinical unit to yield the best fit. [Copyright &y& Elsevier]

Background Brain metastases occur in up to 40% of all patients with systemic cancer. We aimed to assess whether stereotactic radiosurgery provided any therapeutic benefit in a randomised multi-institutional trial directed by the Radiation Therapy Oncology Group (RTOG). Methods Patients with one to three newly diagnosed brain metastases were randomly allocated either whole brain radiation therapy (WBRT) or WBRT followed by stereotactic radiosurgery boost. Patients were stratified by number of metastases and status of extracranial disease. Primary outcome was survival; secondary outcomes were tumour response and local rates, overall intracranial recurrence rates, cause of death, and performance measurements. Findings From January, 1996, to June, 2001, we enrolled 333 patients from 55 participating RTOG institutions--167 were assigned WBRT and stereotactic radiosurgery and 164 were allocated WBRT alone. Univariate analysis showed that there was a survival advantage in the WBRT and stereotactic radiosurgery group for patients with a single brain metastasis (median survival time 6.5 vs 4.9 months, p=0.0393). Patients in the stereotactic surgery group were more likely to have a stable or improved Karnofsky Performance Status (KPS) score at 6 months' follow-up than were patients allocated WBRT alone (43% vs 27%, respectively; p=0.03). By multivariate analysis, survival improved in patients with an RPA class 1 (p<0.0001) or a favourable histological status (p=0.0121). Interpretation WBRT and stereotactic boost treatment improved functional autonomy (KPS) for all patients and survival for patients with a single unresectable brain metastasis. WBRT and stereotactic radiosurgery should, therefore, be standard treatment for patients with a single unresectable brain metastasis and considered for patients with two or three brain metastases. [ABSTRACT FROM AUTHOR]

The β-subunit of the signal recognition particle receptor (SRβ), a member of the Ras family of small molecular weight GTPases, is involved in the targeting of nascent polypeptide chains to the protein translocation machinery in the endoplasmic reticulum membrane. We purified SRβ from an expressing strain of Escherichia coli and investigated the properties of the isolated GTPase. We find that, unlike other Ras family GTPases, most SRβ purifies bound to GTP, and SRβ-bound GTP is not easily exchanged with solution GTP. SRβ possesses no detectable GTPase activity. Although a stable interaction between SRβ and ribosomes is observed, SRβ is not stimulated to hydrolyze GTP when incubated with ribosomes or ribosome-nascent chains. A GTPase mutant harboring a mutation in a region predicted to be functionally important, based on observations made in related GTPases, binds GTP with faster kinetics and appears to be a less stable protein but otherwise displays similar properties to the wild-type SRβ GTPase. Our results demonstrate that as an isolated GTPase, SRβ functions differently from the Arf- and Ras-type GTPases that it is most closely related to by sequence. [ABSTRACT FROM AUTHOR]

The translocation of most proteins across the endoplasmic reticulum or bacterial inner membrane occurs through an aqueous pore that spans the membrane. Substrates that are translocated co-translationally across the membrane are directed to the translocation pore via an interaction between the cytosolic signal recognition particle and its membrane-bound receptor. Together the translocation pore and the receptor are referred to as a translocon. By studying the biogenesis of the translocon a number of alternate targeting and membrane-integration pathways have been discovered that operate independently of the signal recognition particle (SRP) pathway. The novel assembly strategies of the translocon and the ways in which these components interact to ensure the fidelity and unidirectionality of the targeting and translocation process are reviewed here.Key words: protein translocation, translocon, SRP receptor, GTPases.La translocation de la plupart des protéines au travers du réticulum endoplasmique ou de la membrane interne des bactéries se fait par un pore transmembranaire. Les protéines qui passent au travers de la membrane au cours de leur traduction sont dirigées vers le pore de translocation grâce à l'interaction entre la particule de reconnaissance du signal (SRP) dans le cytosol et son récepteur membranaire. Le pore de translocation et le récepteur forment le translocon. En étudiant la biogenèse du translocon, plusieurs voies alternatives de ciblage et d'intégration à la membrane, fonctionnant indépendamment de la voie de la SRP, ont été découvertes. Dans cet article, les nouvelles stratégies d'assemblage du translocon sont passées en revue, ainsi que les mécanismes par lesquels ces constituants interagissent pour assurer la fidélité et le caractère unidirectionnel du processus de ciblage et de translocation.Mots clés : translocation de protéines, translocon, récepteur de la SRP, GTPases.[Traduit par la Rédaction] [ABSTRACT FROM AUTHOR]

Describes the macromolecular transport across cellular membranes. Retrotranslocation of misfolded proteins; Operation of translocation pathways; Recognition of the signal peptide in the cytoplasm; Emergence of a cogent view of protein targeting; Conservation of the translocation of a protein across a membrane between organelles.

Recently, theorists have argued that positive personality dispositions may facilitate the maintenance of satisfying personal relationships. On a sample of 159 couples (married an average of 23.8 years) it was hypothesized that perspective taking. defined as the cognitive tendency to put oneself in another person's place, would be positively predictive of marital adjustment. Three dimensions of perspective taking were measured. A general measure assessed self perspective taking in general social interaction. A second instrument assessed the perspective taking of the self, and the third assessed the perspective taking of another within the marriage relationship. Results indicated that for both husbands and wives, all 3 dimensions of perspective taking were predictive of marital adjustment. Wives had significantly higher scorns than did husbands on all 3 perspective-taking scales. The results are discussed within the context of a model of relational competence. [ABSTRACT FROM AUTHOR]

Presents the results of a detailed biochemical analysis of the functions of the GTPases of SRP and SRP receptor. Discovery of a stable, empty conformation of the GTPase domains; Targeting of nascent secretory polypeptides to the endoplasmic reticulum.

Examines the close friendships of early adolescent boys in relation to antisocial behavior. Similarity between demographic characteristics and antisocial behavior; Correlation between antisocial behavior, directives and negative reciprocity; Implications for clinical and developmental theory.

Suggests ways to intervene and prevent escalation in problem behavior among high-risk adolescents (11-14 years old). Consequences of adolescent problem behavior; Details of a study in this regard; Analysis of results of the study; Limitations.

Presents information on the Adolescent Transitions Project (ATP) program. What is the project; Purpose of the design of the project; Overview of the ATP.

Discusses the possible role of translocons, a site at endoplasmic reticulum, in localization, orientation, maturation and degradation of nascent chains during translation. Components of translocons; Dynamic structure of the translocon; Role of translocon in membrane protein integration.

In the October issue of Molecular Cell, Kowarik et al. examine cotranslational translocation and folding during the synthesis of secretory and integral membrane proteins. Their results suggest that these potentially competing processes are regulated in surprising ways and reveal novel insights into the mechanisms by which proteins are assembled into membranes. [Copyright &y& Elsevier]

Purpose: Owing to their vicinity near the superior sagittal sinus, parasagittal and parafalcine meningiomas are challenging tumors to surgically resect. In this study, we investigate key factors that portend increased risk of recurrence after surgery. Methods: This is a retrospective study of patients who underwent resection of parasagittal and parafalcine meningiomas at our institution between 2012 and 2018. Relevant clinical, radiographic, and histopathological variables were selected for analysis as predictors of tumor recurrence. Results: A total of 110 consecutive subjects (mean age: 59.4 ± 15.2 years, 67.3% female) with 74 parasagittal and 36 parafalcine meningiomas (92 WHO grade 1, 18 WHO grade 2/3), are included in the study. A total of 37 patients (33.6%) exhibited recurrence with median follow-up of 42 months (IQR: 10–71). In the overall cohort, parasagittal meningiomas exhibited shorter progression-free survival compared to parafalcine meningiomas (Kaplan–Meier log-rank p = 0.045). On univariate analysis, predictors of recurrence include WHO grade 2/3 vs. grade 1 tumors (p < 0.001), higher Ki-67 indices (p < 0.001), partial (p = 0.04) or complete sinus invasion (p < 0.001), and subtotal resection (p < 0.001). Multivariable Cox regression analysis revealed high-grade meningiomas (HR: 3.62, 95% CI: 1.60–8.22; p = 0.002), complete sinus invasion (HR: 3.00, 95% CI: 1.16–7.79; p = 0.024), and subtotal resection (HR: 3.10, 95% CI: 1.38–6.96; p = 0.006) as independent factors that portend shorter time to recurrence. Conclusion: This study identifies several pertinent factors that confer increased risk of recurrence after resection of parasagittal and parafalcine meningiomas, which can be used to devise appropriate surgical strategy to achieve improved patient outcomes. [ABSTRACT FROM AUTHOR]

In this issue, Bleicken et al. (2014) use double electron-electron resonance (DEER) spectroscopy to propose a new model for the active form of Bax at membranes that differs significantly from those previously proposed. [ABSTRACT FROM AUTHOR]

Confocal micrographs of EGFP fusion proteins localized at key cell organelles in murine and human cells were acquired for use as subcellular localization landmarks. For each of the respective 789,011 and 523,319 optically validated cell images, morphology and statistical features were measured. Machine learning algorithms using these features permit automated assignment of the localization of other proteins and dyes in both cell types with very high accuracy. Automated assignment of subcellular localizations for model tail-anchored (TA) proteins with randomly mutated C-terminal targeting sequences allowed the discovery of motifs responsible for targeting to mitochondria, endoplasmic reticulum, and the late secretory pathway. Analysis of directed mutants enabled refinement of these motifs and characterization of protein distributions in within cellular subcompartments. [ABSTRACT FROM AUTHOR]

Anti-apoptotic proteins such as BCL-XL promote cell survival by sequestering proapoptotic BCL-2 family members, an activity that frequently contributes to tumorigenesis. Thus, the development of small-molecule inhibitors for anti-apoptotic proteins, termed BH3-mimetics, is revolutionizing how we treat cancer. BH3 mimetics kill cells by displacing sequestered pro-apoptotic proteins to initiate tumor-cell death. Recent evidence has demonstrated that in live cells the BH3- only proteins PUMA and BIM resist displacement by BH3-mimetics, while others like tBID do not. Analysis of the molecular mechanism by which PUMA resists BH3-mimetic mediated displacement from full-length anti-apoptotic proteins (BCL-XL, BCL-2, BCL-W, and MCL-1) reveals that both the BH3-motif and a novel binding site within the carboxyl-terminal sequence (CTS) of PUMA contribute to binding. Together these sequences bind to anti-apoptotic proteins, which effectively ‘double-bolt locks’ the proteins to resist BH3-mimetic displacement. The pro-apoptotic protein BIM has also been shown to double-bolt lock to anti-apoptotic proteins however, the novel binding sequence in PUMA is unrelated to that in the CTS of BIM and functions independent of PUMA binding to membranes. Moreover, contrary to previous reports, we find that when exogenously expressed, the CTS of PUMA directs the protein primarily to the endoplasmic reticulum (ER) rather than mitochondria and that residues I175 and P180 within the CTS are required for both ER localization and BH3-mimetic resistance. Understanding how PUMA resists BH3-mimetic displacement will be useful in designing more efficacious small-molecule inhibitors of anti-apoptotic BCL-2 proteins. [ABSTRACT FROM AUTHOR]

The proapoptotic BCL-2 homology (BH3)-only endoplasmic reticulum (ER)-resident protein BCL-2 interacting killer (BIK) positively regulates mitochondrial outer membrane permeabilization, the point of no return in apoptosis. It is generally accepted that BIK functions at a distance from mitochondria by binding and sequestering antiapoptotic proteins at the ER, thereby promoting ER calcium release. Although BIK is predominantly localized to the ER, we detect by fluorescence lifetime imaging microscopy-FRET microscopy, BH3 region-dependent direct binding between BIK and mitochondria-localized chimeric mutants of the antiapoptotic proteins BCL-XL and BCL-2 in both baby mouse kidney (BMK) and MCF-7 cells. Direct binding was accompanied by cell type-specific differential relocalization in response to coexpression of either BIK or one of its target binding partners, BCL-XL, when coexpressed in cells. In BMK cells with genetic deletion of both BAX and BAK (BMK-double KO), our data suggest that a fraction of BIK protein moves toward mitochondria in response to the expression of a mitochondria-localized BCL-XL mutant. In contrast, in MCF-7 cells, our data suggest that BIK is localized at both ER and mitochondria-associated ER membranes and binds to the mitochondria-localized BCL-XL mutant via relocalization of BCL-XL to ER and mitochondriaassociated ER membrane. Rather than functioning at a distance, our data suggest that BIK initiates mitochondrial outer membrane permeabilization via direct interactions with ER and mitochondria-localized antiapoptotic proteins, which occur via ER-mitochondria contact sites, and/or by relocalization of either BIK or antiapoptotic proteins in cells. [ABSTRACT FROM AUTHOR]

Background: Skull base meningiomas commonly present with cranial neuropathies. Fractionated stereotactic radiation therapy (FSRT) has been used to treat these tumors with excellent local control, but rates of improvement in cranial neuropathies have not been well defined. We review the experience at Thomas Jefferson University using FSRT in the management of these patients with a focus on symptom outcomes.Methods: We identified 225 cases of skull base meningiomas treated with FSRT at Thomas Jefferson University from 1994 through 2009. The target volume was the enhancing tumor, treated to a standard prescription dose of 54 Gy. Symptoms at the time of RT were classified based on the cranial nerve affected. Logistic regression was performed to determine predictors of symptom improvement after FSRT.Results: The median follow-up time was 4.4 years. In 92% of cases, patients were symptomatic at the time of RT; the most common were impaired visual field/acuity (58%) or extraocular movements (34%). After FSRT, durable improvement of at least one symptom occurred in 57% of cases, including 40% of visual acuity/visual field deficits, and 40% of diplopia/ptosis deficits. Of all symptomatic patients, 27% experienced improvement of at least one symptom within 2 months of the end of RT.Conclusions: FSRT is very effective in achieving improvement of cranial neuropathies from skull base meningiomas, particularly visual symptoms. Over half of treated patients experience a durable improvement of at least one symptom, frequently within 2 months from the end of RT. [ABSTRACT FROM AUTHOR]

Early detection and treatment of high-grade dysplasia (HGD) in Barrett's esophagus may reduce the risk of developing esophageal adenocarcinoma. Confocal endomicroscopy (CLE) has shown advantages over routine white-light endoscopic surveillance with biopsy for histological examination; however, CLE is compromised by insufficient contrast and by intra- and interobserver variation. An FDA-approved PDT photosensitizer was used here to reveal morphological and textural features similar to those found in histological analysis. Support vector machines were trained using the aforementioned features to obtain an automatic and robust detection of HGD. Our results showed 95% sensitivity and 87% specificity using the optimal feature combination and demonstrated the potential for extension to a three-dimensional cell model. [ABSTRACT FROM AUTHOR]

High-grade dysplasia (HGD) in Barrett's esophagus (BE) poses increased risk for developing esophageal adenocarcinoma. To date, early detection and treatment of HGD regions are still challenging due to the sampling error from tissue biopsy and relocation error during the treatment after histopathological analysis. In this study, CP-A (metaplasia) and CP-B (HGD) cell lines were used to investigate the "seek-and-treat" potential using 5-aminolevulinic acid-induced protoporphyrin IX (PpIX). The photodynamic therapy photosensitizer then provides both a phototoxic effect and additional image contrast for automatic detection and real-time laser treatment. Complementary to our studies on automatic classification, this work focused on characterizing subcellular irradiation and the potential phototoxicity on both metaplasia and HGD. The treatment results showed that the HGD cells are less viable than metaplastic cells due to more PpIX production at earlier times. Also, due to mitochondrial localization of PpIX, a better killing effect was achieved by involving mitochondria or whole cells compared with just nucleus irradiation in the detected region. With the additional toxicity given by PpIX and potential morphological/textural differences for pattern recognition, this cellular platform serves as a platform to further investigate real-time "seek-and-treat" strategies in three-dimensional models for improving early detection and treatment of BE. [ABSTRACT FROM AUTHOR]

The article presents a study which explores the efficacy and specificity of inhibitors of BCL-2 family protein interactions assessed by affinity measurements in live cells . It mentions that cytoplasmic and membrane-bound BCL-2 family proteins regulate apoptosis, a form of programmed cell death, via dozens of binary protein interactions confounding measurement of the effects of inhibitors in live cells.

The article presents a study which explores the efficacy and specificity of inhibitors of BCL-2 family protein interactions assessed by affinity measurements in live cells . It mentions that cytoplasmic and membrane-bound BCL-2 family proteins regulate apoptosis, a form of programmed cell death, via dozens of binary protein interactions confounding measurement of the effects of inhibitors in live cells.

Introduction: High grade gliomas are an insidious disease associated with an extremely poor prognosis. The role of re-irradiation for recurrent gliomas is unclear but several retrospective studies have indicated mild toxicity and modest outcomes with this regimen. With subsequent progression, it is unclear what options remain and more radiotherapy is rarely offered for fear of surpassing normal central nervous system tissue tolerance and causing significant side effects without significant benefit.Case Presentation: In this report, we describe a 37-year-old Caucasian male initially diagnosed with a grade IV oligodendroglioma, who received multiple courses of re-irradiation and experienced a survival of 10 years with minimal cognitive or neurologic deficits.Conclusion: Significant toxicity with multiple courses of radiation does not always occur. Re-irradiation should be considered in a salvage setting. [ABSTRACT FROM AUTHOR]

Many different signals for cell death converge on outer mitochondrial membrane permeabilization, a process controlled by the Bcl-2 family of proteins. Recent structural and biochemical analyses emphasize that key regulatory interactions between different classes of Bcl-2 family members occur at and in intracellular membranes, and insertion into membranes causes functionally important conformational changes in the proteins. [ABSTRACT FROM AUTHOR]

Although the importance of natural killer (NK) cells in innate immune responses against tumors or viral infections are well documented, their ability to directly recognize pathogens is less well defined. We have recently reported FimH, a bacterial fimbrial protein, as a novel Toll-like receptor (TLR)4 ligand that potently induces antiviral responses. Here, we investigated whether FimH either directly or indirectly can activate human and murine NK cells. We demonstrate that FimH potently activates both human and murine NK cells in vitro to induce cytokines [interferon (IFN)-γ and tumor necrosis factor (TNF)-α] and cytotoxicity. Importantly, NK cells directly recognize FimH-expressing pathogens as FimH+, but not FimH−, bacteria were able to activate human NK cells. FimH activation of NK cells required TLR4 and MyD88 signaling, as NK cells from both TLR4−/− and MyD88−/− mice as well as human NK-92 cells, which lack TLR4, were all unresponsive to FimH. In addition, TLR4 neutralization significantly abrogated the response of human NK cells to FimH. Activation of purified NK cells by FimH was independent of lipopolysaccharide (LPS) or other bacterial contaminations. These data demonstrate for the first time that highly purified NK cells directly recognize and respond to FimH via TLR4–MyD88 pathways to aid innate protection against cancer or microbial infections. [ABSTRACT FROM AUTHOR]

Abstract: Objective: The purpose of this study was to extend the limited research on college student support for alcohol control policies by using a latent class analysis to examine the shared characteristics of drinking students who support or oppose such policies. Methods: We used data from a sample of 2393 students drawn from a larger study on high risk drinking at a mid-western university. Data was collected between October 2005 and May 2007. We conducted a latent class analysis to identify sub-groups of drinking students based on relevant variables. Results: The results of the latent class analysis yielded a model which could correctly classify 90% of the students taking the survey into one of four “classes” based upon their response to four items on the questionnaire. Conclusions: Interventions would benefit from approaches that target both student perceptions and specific policies that are most conducive to student support and engagement. [Copyright &y& Elsevier]

Purpose: To assess the progression patterns in patients with multifocal glioblastoma multiforme who had undergone whole brain radiotherapy (WBRT), the historical standard, versus three-dimensional conformal radiotherapy, and to identify predictive treatment and pretreatment factors. Methods and Materials: The records of 50 patients with multifocal glioblastoma multiforme treated with RT were reviewed. Univariate analyses were performed using survival methods and the Cox proportional hazards regression method. Multivariate analyses were performed using the Cox proportional hazards regression method. Results: The mean age was 61 years, and 71% had a Karnofsky performance status (KPS) score of ≥70. Of the 50 patients, 32% underwent WBRT and 68%, three-dimensional conformal RT. Progression was local in all evaluable patients, as determined by imaging in 38 patients and early neurologic progression in 12. The median time to progression (TTP) was 3.1 months, and the median survival time (MST) was 8.1 months. The significant independent predictors of TTP on multivariate analysis were a KPS score <70 (p = 0.001), the extent of surgery (p = 0.040), a radiation dose <60 Gy (p = 0.027), and the lack of chemotherapy (p = 0.001). The significant independent predictors of a reduced MST were a KPS score <70 (p = 0.022) and the absence of salvage surgery (p = 0.011) and salvage chemotherapy (p = 0.003). Conclusion: Local progression was observed in all patients. On multivariate analysis, no significant difference was found in the TTP or MST between three-dimensional conformal radiotherapy and WBRT. The KPS was a consistent independent predictor of both TTP and MST. On the basis of the progression pattern, we do not recommend WBRT as a mandatory component of the treatment of multifocal glioblastoma multiforme. [Copyright &y& Elsevier]

Cellular adhesion is regulated by members of the cadherin family of adhesion receptors and their cytoplasmic adaptor proteins, the catenins. Adhesion complexes are regulated by recycling from the plasma membrane and proteolysis during apoptosis. We report that in MCF-7, MDA-MB-468 and MDCK cells, induction of apoptosis by agents that cause endoplasmic reticulum (ER) stress results in O-glycosylation of both β-catenin and the E-cadherin cytoplasmic domain. O-glycosylation of newly synthesized E-cadherin blocks cell surface transport, resulting in reduced intercellular adhesion. O-glycosylated E-cadherin still binds to β- and γ-catenin, but not to p120-catenin. Although O-glycosylation can be inhibited with caspase inhibitors, cleavage of caspases associated with the ER or Golgi complex does not correlate with E-cadherin O-glycosylation. However, agents that induce apoptosis via mitochondria do not lead to E-cadherin O-glycosylation, and decrease adhesion more slowly. In MCF-7 cells, this is due to degradation of E-cadherin concomitant with cleavage of caspase-7 and its substrate poly(ADP-ribose) polymerase. We conclude that cytoplasmic O-glycosylation is a novel, rapid mechanism for regulating cell surface transport exploited to down-regulate adhesion in some but not all apoptosis pathways. [ABSTRACT FROM AUTHOR]

Bax proteins form pores in the mitochondrial outer membrane to initiate apoptosis. This might involve their embedding in the cytosolic leaflet of the lipid bilayer, thus generating tension to induce a lipid pore with radially arranged lipids forming the wall. Alternatively, Bax proteins might comprise part of the pore wall. However, there is no unambiguous structural evidence for either hypothesis. Using NMR, we determined a high‐resolution structure of the Bax core region, revealing a dimer with the nonpolar surface covering the lipid bilayer edge and the polar surface exposed to water. The dimer tilts from the bilayer normal, not only maximizing nonpolar interactions with lipid tails but also creating polar interactions between charged residues and lipid heads. Structure‐guided mutations demonstrate the importance of both types of protein–lipid interactions in Bax pore assembly and core dimer configuration. Therefore, the Bax core dimer forms part of the proteolipid pore wall to permeabilize mitochondria. Synopsis: Bax proteins form giant pores in mitochondrial membranes to initiate cell death. Structural determination and activity analyses of membrane‐bound amphipathic Bax core dimers disambiguate their function as building blocks of the apoptotic pore wall. High‐resolution NMR analyses reveal the structure of the α2–α5 helices of Bax bound to a lipid bilayer.This Bax core region forms a symmetric dimer interacting with lipid bilayer with a 60° tilt angle, to cover both the nonpolar acyl tails and the polar phosphate heads.Both nonpolar and polar lipid contacts of the Bax core dimer dictate its configuration and pore forming activity in model bilayers as well as native mitochondrial membranes. [ABSTRACT FROM AUTHOR]

Patients with grade III anaplastic astrocytomas (AA) separate into survival cohorts based on the presence or absence of mutations in isocitrate dehydrogenase (IDH). Progression to glioblastoma (GBM), morphologically distinguishable by elevated microvascular proliferation, necrosis, and cell division in tumor tissues, is considerably more rapid in IDH wild-type tumors such that their diagnosis as AA is relatively rare. More often initially presenting as GBM, these contain higher numbers of tumor-associated macrophages (TAMs) than most AA, and GBM patients also have higher levels of circulating M2 monocytes. TAM and M2 monocytes share functional properties inhibitory for antitumor immunity. Yet, although there is a wealth of data implicating TAM in tumor-immune evasion, there has been limited analysis of the impact of the circulating M2 monocytes. In the current study, immune parameters in sera, circulating cells, and tumor tissues from patients with primary gliomas morphologically diagnosed as AA were assessed. Profound differences in serum cytokines, glioma extracellular vesicle cross-reactive Abs, and gene expression by circulating cells identified two distinct patient cohorts. Evidence of type 2-immune bias was most often seen in patients with IDH wild-type AA, whereas a type 1 bias was common in patients with tumors expressing the IDH1R132H mutation. Nevertheless, a patient's immune profile was better correlated with the extent of tumor vascular enhancement on magnetic resonance imaging than IDH mutational status. Regardless of IDH genotype, AA progression appears to be associated with a switch in systemic immune bias from type 1 to type 2 and the loss of tumor vasculature integrity. [ABSTRACT FROM AUTHOR]

Phenotypic profiling of large three-dimensional microscopy data sets has not been widely adopted due to the challenges posed by cell segmentation and feature selection. The computational demands of automated processing further limit analysis of hard-to-segment images such as of neurons and organoids. Here we describe a comprehensive shallow-learning framework for automated quantitative phenotyping of three-dimensional (3D) image data; using unsupervised data-driven voxel-based feature learning, which enables computationally facile classification, clustering and advanced data visualization. We demonstrate the analysis potential on complex 3D images by investigating the phenotypic alterations of: neurons in response to apoptosis-inducing treatments and morphogenesis for oncogene-expressing human mammary gland acinar organoids. Our novel implementation of image analysis algorithms called Phindr3D allowed rapid implementation of data-driven voxel-based feature learning into 3D high content analysis (HCA) operations and constitutes a major practical advance as the computed assignments represent the biology while preserving the heterogeneity of the underlying data. Phindr3D is provided as Matlab code and as a stand-alone program (https://github.com/DWALab/Phindr3D). Author summary: Fluorescence microscopy is a fundamental technology for cell biology. However, unbiased quantitative phenotypic analysis of microscopy images of cells grown in 3D organoids or in dense culture conditions in large enough numbers to reach statistical clarity remains a fundamental challenge. Here, we report that using data driven voxel-based features and machine learning it is possible to analyze complex 3D image data without compressing them to 2D, identifying individual cells or using computationally intensive deep learning techniques. Further, we present methods for analyzing this data by classification or clustering. Together these techniques provide the means for facile discovery and interpretation of meaningful patterns in a high dimensional feature space without complex image processing and prior knowledge or assumptions about the feature space. Our method enables novel opportunities for rapid large-scale multivariate phenotypic microscopy image analysis in 3D using a standard desktop computer. [ABSTRACT FROM AUTHOR]

The Bcl-2 family BH3 protein Bim promotes apoptosis at mitochondria by activating the pore-forming proteins Bax and Bak and by inhibiting the anti-apoptotic proteins Bcl-XL, Bcl-2 and Mcl-1. Bim binds to these proteins via its BH3 domain and to the mitochondrial membrane by a carboxyl-terminal sequence (CTS). In cells killed by Bim, the expression of a Bim mutant in which the CTS was deleted (BimL-dCTS) triggered apoptosis that correlated with inhibition of anti-apoptotic proteins being sufficient to permeabilize mitochondria isolated from the same cells. Detailed analysis of the molecular mechanism demonstrated that BimL-dCTS inhibited Bcl-XL but did not activate Bax. Examination of additional point mutants unexpectedly revealed that the CTS of Bim directly interacts with Bax, is required for physiological concentrations of Bim to activate Bax and that different residues in the CTS enable Bax activation and binding to membranes. [ABSTRACT FROM AUTHOR]

Background: Tail-anchored membrane proteins (TAMPs) differ from other integral membrane proteins, because they contain a single transmembrane domain at the extreme carboxyl-terminus and are therefore obliged to target to membranes post-translationally. Although 3–5% of all transmembrane proteins are predicted to be TAMPs only a small number are well characterized. Results: To identify novel putative TAMPs across different species, we used TAMPfinder software to identify 859, 657 and 119 putative TAMPs in human (Homo sapiens), plant (Arabidopsis thaliana), and yeast (Saccharomyces cerevisiae), respectively. Bioinformatics analyses of these putative TAMP sequences suggest that the list is highly enriched for authentic TAMPs. To experimentally validate the software predictions several human and plant proteins identified by TAMPfinder that were previously uncharacterized were expressed in cells and visualized at subcellular membranes by fluorescence microscopy and further analyzed by carbonate extraction or by bimolecular fluorescence complementation. With the exception of the pro-apoptotic protein harakiri, which is, peripherally bound to the membrane this subset of novel proteins behave like genuine TAMPs. Comprehensive bioinformatics analysis of the generated TAMP datasets revealed previously unappreciated common and species-specific features such as the unusual size distribution of and the propensity of TAMP proteins to be part of larger complexes. Additionally, novel features of the amino acid sequences that anchor TAMPs to membranes were also revealed. Conclusions: The findings in this study more than double the number of predicted annotated TAMPs and provide new insights into the common and species-specific features of TAMPs. Furthermore, the list of TAMPs and annotations provide a resource for further investigation. [ABSTRACT FROM AUTHOR]

Tail-anchored (TA) proteins are a unique class of integral membrane proteins that possess a single C-terminal transmembrane domain and target post-translationally to the specific organelles at which they function. While significant advances have been made in recent years in elucidating the mechanisms and molecular targeting signals involved in the proper sorting of TA proteins, particularly to the endoplasmic reticulum and mitochondria, relatively little is known about the targeting of TA proteins to the plastid outer envelope. Here we show that several known or predicted plastid TA outer envelope proteins (OEPs) in Arabidopsis possess a C-terminal RK/ST sequence motif that serves as a conserved element of their plastid targeting signal. Evidence for this conclusion comes primarily from experiments with OEP7.2, which is a member of the Arabidopsis 7 kDa OEP family. We confirmed that OEP7.2 is localized to the plastid outer envelope and possesses a TA topology, and its C-terminal sequence (CTS), which includes the RK/ST motif, is essential for proper targeting to plastids. The CTS of OEP7.2 is functionally interchangeable with the CTSs of other TA OEPs that possess similar RK/ST motifs, but not with those that lack the motif. Further, a bioinformatics search based on a consensus sequence led to the identification of several new OEP TA proteins. Collectively, this study provides new insight into the mechanisms of TA protein sorting in plant cells, defines a new targeting signal element for a subset of TA OEPs and expands the number and repertoire of TA proteins at the plastid outer envelope. [ABSTRACT FROM AUTHOR]