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Intravenous (IV) recombinant tissue plasminogen activator is the standard of care for patients with acute ischemic stroke (AIS) who present to the hospital within 4.5 hours of symptom onset. However, IV thrombolysis, even bridging thrombolysis (combining intravenous thrombolysis and mechanical thrombectomy) has limited efficacy among patients who had occlusive lesions associated with high-grade arterial stenosis requiring revascularization to improve neurological deficits. We evaluated whether rescue stenting results in good outcomes among patients after the failure of intravenous thrombolysis and bridging thrombolysis.We retrospectively analyzed patients with AIS who underwent rescue stenting for large vessel occlusion with severe atherosclerotic stenosis between May 2020 and August 2022 at Can Tho S.I.S General Hospital. Primary outcomes included the incidence of hemorrhagic transformation and the rate of good outcomes (modified Rankin Scale3) at 3-month follow-up.We identified 13 patients who received rescue stenting after the failure of IV alteplase and bridging thrombolysis, but only 11 patients met the inclusion criteria. All patients experienced successful recanalization, and 1 (9.1%) patient experienced new infarcts. Of these 11 patients, 10 (90.9%) had good outcomes 3 months after rescue stenting. Additionally, a loading dose of dual antiplatelet therapy (DAPT) applied concurrently with IV alteplase improved the recanalization rate for large target arteries but had no significant effect on the incidence of symptomatic intracranial hemorrhage.Rescue stenting appears to represent an additional therapeutic option in cases that fail to resolve with IV alteplase, which may improve clinical outcomes.

In cases of acute ischemic stroke (AIS) caused by intracranial large vessel occlusion, rescue intracranial stenting (RIS) has recently emerged as a treatment option for achieving recanalization when mechanical thrombectomy (MT) fails. However, few studies to date have reported on the beneficial outcomes of RIS. Our goal was to analyze whether RIS use can improve prognosis in patients 3 months post-treatment.A retrospective analysis was performed on a prospective cohort of patients with AIS treated with RIS at Can Tho S.I.S General Hospital. The study inclusion criteria were evidence of intracranial large vessel occlusion, absence of intracranial hemorrhage (ICH), and severe stenosis or reocclusion after MT. Patients with tandem occlusions, failure to follow up after discharge, or severe or fatal illness concomitant with AIS were excluded from the study. The primary outcome was the "non-poor" prognosis status rate at 3 months after RIS and post-procedural symptomatic ICH (sICH).The post-treatment outcomes of 85 eligible patients who received RIS between August 2019 and May 2021 were assessed. Of the 85 included patients, 82 (96.5%) achieved successful recanalization, and 4 (4.7%) experienced sICH. At 3-months post-treatment, 47 (55.3%) patients had "non-poor" outcomes, whereas 35 (41.2%) had good outcomes. The use of dual antiplatelet therapy was associated with new infarcts (relative risk [RR]: 0.1; 95% confidence interval [CI]: 0.01-0.7) and sICH occurrence (RR: 0.1; 95% CI: 0.01-0.9).Our study suggests that despite the occurrence of post-procedural sICH in a small proportion of cases, RIS could serve as a useful alternative or additional treatment in the event of MT failure.

Autoantibodies targeting brain antigens can mediate a wide range of neurological symptoms ranging from epileptic seizures to psychosis to dementia. Although earlier experimental work indicated that autoantibodies can be directly pathogenic, detailed studies on disease mechanisms, biophysical autoantibody properties, and target interactions were hampered by the availability of human material and the paucity of monospecific disease-related autoantibodies. The emerging generation of patient-derived monoclonal autoantibodies (mAbs) provides a novel platform for the detailed characterization of immunobiology and autoantibody pathogenicity in vitro and in animal models. This Feature Review focuses on recent advances in mAb generation and discusses their potential as powerful scientific tools for high-resolution imaging, antigenic target identification, atomic-level structural analyses, and the development of antibody-selective immunotherapies.

Immuncheckpointinhibitoren (ICI) sind bei zahlreichen Tumorentitaten wirksam, konnen allerdings immunvermittelte Nebenwirkungen in allen Organsystemen induzieren und so zu langfristigen Schaden und Todesfallen fuhren. Hierbei konnen Nebenwirkungen auch Monate nach Beendigung der Therapie auftreten und Spatfolgen Jahre nach Beendigung der Therapie bestehen bleiben. Aufgrund des zunehmenden Einsatzes von ICI auch in fruheren Tumorstadien mussen diese langfristigen Auswirkungen in die Nutzen-Risiko-Bewertung miteinbezogen werden. Es wird eine Ubersicht uber die Spatfolgen einer ICI-Therapie gegeben sowie die Datenbasis in Form des internationalen Side Effect Registry Immuno-Oncology (SERIO) zur Dokumentation und Auswertung der immunvermittelten Nebenwirkungen vorgestellt. Neben der Auswertung des SERIO-Registers erfolgte eine selektive Literaturrecherche in den Datenbanken PubMed, Cochrane Central Register of Controlled Trials und Google Scholar. Die haufigsten immunvermittelten Nebenwirkungen mit langfristigen Folgen sind kutan (Leukotrichie und melanomassoziierte Hypopigmentierung) und endokrin (Thyreoiditis, Hypophysitis, Adrenalitis, Diabetes). Wahrend Erstere v. a. als stigmatisierend erlebt werden, wird bei den endokrinen Nebenwirkungen meist eine lebenslange Hormonsubstitution notwendig. Neurologische und kardiologische immunvermittelte Nebenwirkungen sind zwar selten, jedoch mit hoher Morbiditat und Mortalitat assoziiert. Bisher sind prospektive Daten zu langfristigen Auswirkungen von immunvermittelten Nebenwirkungen nicht verfugbar. Das SERIO-Register bietet die Moglichkeit, klinische Falle systematisch zu sammeln, um ICI-assoziierte Nebenwirkungen besser zu charakterisieren und nachzuverfolgen.

Immune checkpoint inhibitors (ICIs) have emerged as a promising class of cancer immunotherapies. Neurotoxicities are uncommon, but often severe, and potentially fatal complications of ICIs, and clinical experience is limited. The aim of this study is to further define the clinical spectrum and outcome of ICI-mediated neurotoxicities. Patients with ICI-associated neurotoxicities were identified from retrospective review of the quality control database at a single institution. Data regarding demographics, medical history, clinical presentation, diagnosis, management and outcome were recorded. We identified 18 patients with neurotoxicity following ICI therapy with pembrolizumab, nivolumab, atezolizumab, or ipilimumab for a diverse set of malignancies. Neurotoxicities comprised central demyelinating disorder (28%), autoimmune encephalitis predominantly affecting the grey matter (17%), aseptic meningitis (6%), myasthenia gravis (MG) (17%) with concurrent myositis (6%), sensorimotor polyneuropathy (11%) and hypophysitis (17%). Median time to onset of neurotoxicities was 5 weeks (range 1–72). All patients discontinued ICIs and received steroids with additional immunomodulation required in 9 patients, resulting in improvement for 16 of 18 patients. Grade 3–4 neurotoxicity developed in 14 patients, of whom 6 had died at database closure. Grade 3–4 severity negatively impacted overall survival (OS) (p = 0.046). ICI-mediated neurotoxicities present early, are rapidly progressive and include a diverse phenotype affecting the CNS, PNS and neuroendocrine system. A high level of vigilance is warranted, as early diagnosis and targeted treatment can substantially prevent morbidity and mortality. Prospective clinical trials are warranted to assess optimized management of ICI-induced neurotoxicities.

Background: Polycomb repressive complex 2 (PRC2) tri-methylates histone H3 at lysine 27 (H3K27me3) leading to transcriptionally silenced genes. ORIC-944 is a potent, highly selective allosteric small molecule inhibitor of PRC2’s embryonic ectoderm development (EED) subunit that is undergoing clinical development as an orally bioavailable monotherapy in patients with metastatic prostate cancer. We previously reported strong tumor growth inhibition with ORIC-944 in enzalutamide-resistant 22Rv1 prostate cancer xenografts. Here, to devise and implement a biomarker strategy aimed at ORIC-944 dose selection in the ongoing phase 1 study (NCT05413421), we undertook preclinical pharmacology studies and assay development. Methods: Naïve and 22Rv1-tumor bearing male mice were treated with vehicle, 10, 30 or 100 mg/kg ORIC-944 PO for up to 37 days. H3K27me3 was evaluated in dorsal skin (epidermal layer) and peripheral monocytes by IHC and an alphaLISA test, respectively. Since dying tumor cells release nucleosomes to circulation, cell-free (cf)-nucleosomal H3K27me3 levels normalized to cf-H3.1 were also assessed in plasma using a proprietary magnetic bead-based sandwich immunoassay. Tumors were profiled by RNA-sequencing and H3K27me3 chromatin immunoprecipitation sequencing. Results: In the epidermal skin layer, H3K27me3 was present in vehicle-treated naïve animals but strongly depleted in ORIC-944-treated mice in a dose-dependent manner. Likewise in monocytes, H3K27me3 was reduced to undetectable levels in 100 mg/kg ORIC-944-treated mice. In plasma, normalized cf-nucleosomal H3K27me3 levels significantly decreased in response to ORIC-944 in a dose- and time-dependent manner, selectively in 22Rv1-tumor bearing mice compared to non-tumor bearing mice. Putative PRC2 target genes were identified in 22Rv1 xenografts by focusing on those genes with H3K27me3 binding in the promoter, whose expression was de-repressed by ORIC-944 in a time-dependent manner and then re-expressed after dose suspension. In the ongoing phase 1 study, H3K27me3 levels are being evaluated in the stratum spinosum of skin and in blood-derived monocytes using a proprietary IHC assay and alphaLISA test, respectively. Modulation in the expression of the putative PRC2 target genes is being assessed in blood-derived peripheral blood mononuclear cells by RNA-sequencing. Conclusion: This comprehensive biomarker strategy enables us to establish target engagement and capture exposure-dependent pharmacodynamics in the ongoing phase 1 study evaluating ORIC-944 as a best-in-class PRC2 inhibitor for the treatment of patients with advanced prostate cancer. Citation Format: Anneleen Daemen, Natalie Yuen, Aleksandr Pankov, Eric A. Ariazi, Subhash D. Katewa, Frank L. Duong, Amber Wang, Shravani Barkund, Shelly Kaushik, Jessica D. Sun, Lori S. Friedman, Melissa R. Junttila. Biomarker strategy for a phase 1 study of ORIC-944, a potent and selective allosteric PRC2 inhibitor, in patients with metastatic prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2791.

The neuropeptide calcitonin gene-related peptide (CGRP) is a major player in migraine pathophysiology. Previous preclinical studies demonstrated that intracerebroventricular administration of CGRP caused migraine-like behaviors in mice, but the sites of action in the brain remain unidentified. The cerebellum has the most CGRP binding sites in the central nervous system and is increasingly recognized as both a sensory and motor integration center. The objective of this study was to test whether the cerebellum, particularly the medial cerebellar nuclei (MN), might be a site of CGRP action. In this study, CGRP was directly injected into the right MN of C57BL/6J mice via a cannula. A battery of behavioral tests was done to assess migraine-like behaviors. CGRP caused light aversion measured as decreased time in the light zone even with dim light. The mice also spent more time resting in the dark zone, but not the light, along with decreased rearing and transitions between zones. These behaviors were similar for both sexes. In contrast, significant responses to CGRP were seen only with female mice in the open field assay, von Frey test, and automated squint assay, indicating anxiety, tactile hypersensitivity, and spontaneous pain, respectively. In male mice, the responses had the same trend as females but did not reach statistical significance. No detectable effect of CGRP on gait was observed in either sex. These results suggest that CGRP in the MN causes light aversion in males, while in females, light aversion is accompanied by increased anxiety, tactile hypersensitivity, and spontaneous pain. A caveat is that we cannot exclude contributions from other cerebellar regions in addition to the MN due to diffusion of the injected peptide. These results reveal the cerebellum as a new site of CGRP actions that may contribute to migraine pathophysiology and possibly its prevalence in females.

The introduction of immune checkpoint inhibitors (ICIs) in oncologic therapies has led to a paradigm shift in cancer treatment. ICIs have increased the overall survival in patients with malignant melanoma, small-cell lung cancer, and many other tumor entities. Despite their clinical benefits, these novel cancer immunotherapies can induce neurological immune-related adverse events (irAEs). Such immune-mediated complications can manifest within the spectrum of paraneoplastic neurological syndromes (PNSs). PNSs are rare immune-mediated complications of systemic cancers that can involve every aspect of the nervous system. The emergence of PNSs with ICI treatment opens further pathways to study the complex immunopathological interplay of cancer immunity, cross-reactive neurological autoimmune phenomena, and effects of ICIs on the immune system. ICI-induced PNSs comprise a diverse antibody repertoire and phenotypic spectrum with severe and life-threatening disease progression in some cases. Timely diagnosis and urgent interventions are pivotal for a favorable neurologic and oncologic outcome. This review focuses on the pathogenesis of cancer immunotherapy and the disruption of immune tolerance in PNSs and provides an overview of the most pertinent clinical manifestations and principles of diagnostic and therapeutic managements in light of the expected increase in PNSs due to the widespread use of ICIs in clinical practice. This review further discusses potential and evolving concepts of therapeutic monoclonal antibodies for the treatment of PNSs.

In the crystalline state, salicyl­aldehyde N(4)-morpholino­thio­semicarbazone forms sheets parallel to (002) and consisting of two parallel chains running in the a-axis direction and formed by N—H⋯O and C—H⋯O hydrogen bonds., The structure of the title compound (systematic name: N-{[(2-hy­droxy­phen­yl)methyl­idene]amino}­morpholine-4-carbo­thio­amide), C12H15N3O2S, was prev­iously determined (Koo et al., 1977 ▸) using multiple-film equi-inclination Weissenberg data, but has been redetermined with higher precision to explore its conformation and the hydrogen-bonding patterns and supra­molecular inter­actions. The mol­ecular structure shows intra­molecular O—H⋯N and C—H⋯S inter­actions. The configuration of the C=N bond is E. The mol­ecule is slightly twisted about the central N—N bond. The best planes through the phenyl ring and the morpholino ring make an angle of 43.44 (17)°. In the crystal, the mol­ecules are connected into chains by N—H⋯O and C—H⋯O hydrogen bonds, which combine to generate sheets lying parallel to (002). The most prominent contribution to the surface contacts are H⋯H contacts (51.6%), as concluded from a Hirshfeld surface analysis.

The title compound, C30H23KO8S2, contains one mol­ecule of (E)-2-{4-[3-(thio­phen-3-yl)acrylo­yl]phen­oxy}acetic acid and one mol­ecule of its potassium salt in the asymmetric unit. The distorted KO6 octa­hedra share edges, resulting in chains running in the [010] direction., The synthesis and spectroscopic data of (E)-2-{4-[3-(thio­phen-3-yl)acrylo­yl]phen­oxy}acetic acid are described. Crystallization from an ethanol–water mixture resulted in the title compound, C30H23KO8S2 or [K(C15H11O4S)(C15H12O4S)]n, containing one mol­ecule of the acid and one mol­ecule of the potassium salt in the asymmetric unit. Both mol­ecules share the H atom between their carboxyl groups and a potassium ion. The C=C bonds display an E configuration. The thio­phene and phenyl rings in the two mol­ecules are inclined by 43.3 (2) and 22.7 (2)°. The potassium ion is octa­hedrally coordinated by six O atoms. This distorted octa­hedron shares on opposite sides two oxygen atoms with inversion-related octa­hedra, resulting in chains of octa­hedra running in the [010] direction, which form ladder-like chains by C—H⋯π inter­actions. A Hirshfeld surface analysis indicates that the highest contributions to the surface contacts arise from inter­actions in which H atoms are involved, with the most important contribution being from H⋯H (31.6 and 31.9% for the two mol­ecules) inter­actions.

International audience; Background and purpose: Better understanding the incidence, predictors and mechanisms of early neurological deterioration (END) following intravenous thrombolysis (IVT) for acute stroke with mild symptoms and isolated internal carotid artery occlusion (iICAo) may inform therapeutic decisions.Methods: From a multicenter retrospective database, we extracted all patients with both National Institutes of Health Stroke Scale (NIHSS) score

Finding the Optimal Power Flow (OPF) which minimizes total generator cost is the answer to one of the most important problems in the electricity market operation. Independent System Operators (ISO) face many challenges while operating the system in order to obtain economic benefits and security. The solution to this problem is known as Security Constrained Optimal Power Flow (SCOPF). SCOPF is a very large-scale and nonlinear optimization problem with many complex constraints. This paper proposes the Sunflower Optimization (SFO) algorithm for solving the SCOPF problem. The proposed method is tested on the IEEE 30-bus and the IEEE 118-bus systems for both normal and outage cases. The result comparison with other known methods showed that the proposed SFO algorithm is an effective method for solving the SCOPF problem in the electricity market.

Background: Anterior cruciate ligament (ACL) reconstruction can effectively return athletes to the playing field, but they are still at risk of developing posttraumatic osteoarthritis (PTOA). No studies have used multivariable analysis to evaluate the predictors of radiographic PTOA in the lateral compartment of the knee at short-term follow-up after ACL reconstruction. Purpose: To determine the predictors of radiographic joint space narrowing in the lateral compartment 2 to 3 years after ACL reconstruction in a young, active cohort. Study Design: Cohort study; Level of evidence, 2. Methods: A nested cohort of 358 patients from the Multicenter Orthopaedic Outcomes Network (MOON) prospective cohort who were aged ≤33 years, were injured playing a sport, and had never undergone surgery on the contralateral knee were followed up 2 years after ACL reconstruction with questionnaires and with weightbearing knee radiographs using the metatarsophalangeal (MTP) joint technique. The joint space width in the lateral compartment was measured using a semiautomatic computerized method, and multivariable predictive modeling was used to evaluate the relationship between meniscus treatment, cartilage injury, graft type, and joint space while adjusting for age, sex, body mass index, and Marx activity score. Results: The mean lateral joint space width was 0.11 mm narrower on the ACL-reconstructed knee compared with the contralateral healthy knee (7.69 mm vs 7.80 mm, respectively; P < .01). Statistically significant predictors of a narrower joint space width on the ACL-reconstructed knee included lateral meniscectomy ( P < .001) and a Marx activity score less than 16 points ( P < .001). Conclusion: This study identifies lateral meniscectomy and a lower baseline Marx activity score to be predictors of radiographic joint space narrowing in the lateral compartment 2 to 3 years after ACL reconstruction in young, active patients without a prior knee injury.

This work aims to characterize the relationship between tear film neuropeptide substance P and the structural integrity of the sub-basal nerve plexus in diabetes.Seventeen healthy control participants and nine participants with diabetes were recruited in this cross-sectional study. Total protein content and substance P concentrations were determined in the flush tears of participants. Corneal nerve morphology was assessed by capturing the corneal sub-basal nerve plexus using the Heidelberg Retinal Tomograph II with the Rostock Corneal Module (Heidelberg Engineering GmbH, Heidelberg, Germany) in the central cornea. Corneal nerve fiber density (CNFD) was measured using ACCMetrics (M.A. Dabbah, Imaging Science and Biomedical Engineering, Manchester, UK) on eight captured images. Comparisons between groups were made using independent samples t-tests. Correlations between parameters were analyzed using Pearson's correlations.Substance P concentrations were significantly higher in the tears of the control group compared to participants with diabetes (4150 ± 4752 and 1473 ± 1671 pg/mL, respectively, P = .047). There was no significant difference in total protein content between the groups (3.4 ± 1.8 and 2.6 ± 1.7 mg/mL in the control and diabetes groups, respectively, P = .262). CNFD was significantly lower in the participants with diabetes compared to the control group (16.1 ± 5.7 and 21.5 ± 7.0 mm/mm, respectively, P = .041). There was a moderate correlation between substance P and CNFD (r = 0.48, P = .01).Substance P is expressed at a significantly lower level in the tears of people with diabetes compared with healthy controls. The positive correlation between substance P and corneal nerve density indicates that substance P may be a potential biomarker for corneal nerve health.

The dopamine transporter, also known as solute carrier 6A3 (SLC6A3), plays an important role in synaptic transmission by regulating the reuptake of dopamine in the synapses. In line with this, variations in the gene encoding this transporter have been linked to both schizophrenia and affective disorders. Recently, copy number variants (CNVs) in SLC6A3 have been identified in healthy subjects but so far, the implication of CNVs affecting this gene in psychiatric diseases has not been addressed.In the present study, we aimed to investigate whether CNVs affecting SLC6A3 represent rare high-risk variants of psychiatric disorders.We performed a systematic screening for CNVs affecting SLC6A3 in 761 healthy controls, 672 schizophrenia patients, and 194 patients with bipolar disorder in addition to 253 family members from six large pedigrees affected by mental disorders using single nucleotide polymorphism arrays and subsequent verification by real-time polymerase chain reaction.We identified two duplications and one deletion affecting SLC6A3 in the patients, while no such CNVs were identified in any of the controls. The identified CNVs were of different sizes and two affected several genes in addition to SLC6A3.Our findings suggest that rare high-risk CNVs affecting the gene encoding the dopamine transporter contribute to the pathogenesis of schizophrenia and affective disorders.

The purpose of this investigation was to explore the presumed relationship between the days of hospitalisation and microorganisms identified by endotracheal aspirate cultures in relation to adequate empirical treatment strategies of pneumonia in the intensive care unit (ICU). All potentially pathogenic microorganisms identified by (surveillance) cultures of endotracheal aspirates obtained in the ICUs of two Dutch teaching hospitals in 2007 and 2012 were retrospectively collected and analysed. Antibiotic susceptibilities to 11 antibiotics were calculated for several time points (days or weeks) after hospital admission and expressed per patient-day. In total, 4184 potentially pathogenic microorganisms identified in 782 patients were analysed. Prevalence of the classic early-onset pneumonia-causing microorganisms decreased from 55 % on the first four days to 34 % on days 4–6 after hospital admission (p

SummaryObjectiveTo identify risk factors for radiographic signs of post-traumatic osteoarthritis (OA) 2–3 years after anterior cruciate ligament (ACL) reconstruction through multivariable analysis of minimum joint space width (mJSW) differences in a specially designed nested cohort.MethodsA nested cohort within the Multicenter Orthopaedic Outcomes Network (MOON) cohort included 262 patients (148 females, average age 20) injured in sport who underwent ACL reconstruction in a previously uninjured knee, were 35 or younger, and did not have ACL revision or contralateral knee surgery. mJSW on semi-flexed radiographs was measured in the medial compartment using a validated computerized method. A multivariable generalized linear model was constructed to assess mJSW difference between the ACL reconstructed and contralateral control knees while adjusting for potential confounding factors.ResultsUnexpectedly, we found the mean mJSW was 0.35 mm wider in ACL reconstructed than in control knees (5.06 mm (95% CI 4.96–5.15 mm) vs 4.71 mm (95% CI 4.62–4.80 mm), P

A good organization of management accounting, particularly the preparation of production cost estimation, is a critical condition to provide an appropriate, useful and timely information of the cost for the decision making of the managers. Through a review of cost estimation preparation status of enterprises, this essay presents three procedures for making cost estimation by senior-level management, grassroots-level and the module in which these two procedures are combined.

Hybrid zones may serve as bridges permitting gene flow between species, including alleles influencing the evolution of breeding systems. Using greenhouse crosses, we assessed the likelihood that a hybrid zone could serve as a conduit for transfer of nuclear male-sterility alleles between a gynodioecious species and a hermaphroditic species with very rare females in some populations. Segregation patterns in progeny of crosses between rare females of hermaphroditic Schiedea menziesii and hermaphroditic plants of gynodioecious Schiedea salicaria heterozygous at the male-sterility locus, and between female S. salicaria and hermaphroditic plants from the hybrid zone, were used to determine whether male-sterility was controlled at the same locus in the parental species and the hybrid zone. Segregations of females and hermaphrodites in approximately equal ratios from many of the crosses indicate that the same nuclear male-sterility allele occurs in the parent species and the hybrid zone. These rare male-sterility alleles in S. menziesii may result from gene flow from S. salicaria through the hybrid zone, presumably facilitated by wind pollination in S. salicaria. Alternatively, rare male-sterility alleles might result from a reversal from gynodioecy to hermaphroditism in S. menziesii, or possibly de novo evolution of male sterility. Phylogenetic analysis indicates that some species of Schiedea have probably evolved separate sexes independently, but not in the lineage containing S. salicaria and S. menziesii. High levels of selfing and expression of strong inbreeding depression in S. menziesii, which together should favour females in populations, argue against a reversal from gynodioecy to hermaphroditism in S. menziesii.

The emerging invasive fungal pathogen Aspergillus fumigatus causes very serious infections among immunocompromised patient populations. While the genome of this pathogen has been sequenced, a major barrier to better understanding the complex biology of this eukaryotic organism is a lack of tools for efficient genetic manipulation. To improve upon this, we have generated a new gene deletion system for A. fumigatus using yeast recombinational cloning and Agrobacterium tumefaciens mediated transformation (ATMT) employing a recyclable marker system. This system reduced the time for generating a gene deletion strain in our hands by two-thirds (12 weeks to 3 weeks) using minimal human labor, and we demonstrate that it can be used to efficiently generate multiple gene deletions within a single strain.

We present a fiber optic Raman probe capable of distinguishing human prostate tissue types. Preliminary work on ex-vivo human prostates is presented along with work towards live classification and clinical integration.

Clinical recommendations limit menopausal hormone therapy to a few years, yet the impact of a shorter treatment duration on cardiovascular health is unknown. We hypothesized that both short- and long-term estradiol (E2) treatment exerts positive and lasting effects on blood pressure, vascular reactivity, and renal health. This study was designed to mimic midlife menopause, followed by E2 treatment, that either followed or exceeded the current clinical recommendations. Female Long-Evans retired breeders were ovariectomized (OVX) at 11 mo of age and randomized into three groups: 80-day (80d) vehicle (Veh>Veh), 40-day (40d) E2 + 40d vehicle (E2>Veh), and 80d E2 (E2>E2). In comparison to Veh>Veh, both the E2>Veh and E2>E2 groups had lower systolic blood pressure and enhanced mesenteric relaxation in response to estrogen receptor-α stimulation. Despite the reduced blood pressure, E2>E2 induced renal and cardiac hypertrophy, reduced glomerular filtration, and increased proteinuria. Interestingly, kidneys from E2>Veh rats had significantly fewer tubular casts than both of the other groups. In conclusion, long-term E2 lowered blood pressure but exerted detrimental effects on kidney health in midlife OVX Long-Evans rats, whereas short-term E2 lowered blood pressure and reduced renal damage. These findings highlight that the duration of hormone therapy may be an important factor for renal health in aging postmenopausal women.

Menopause accelerates the development of hypertension, arterial stiffness, end organ damage, and diastolic dysfunction. Postmenopausal hormone therapy relieves menopausal symptoms but promotes adverse cardiovascular outcomes, which may be due to aging-induced alterations in estrogen receptors. We previously published that GPER expression as well as agonist-induced vasorelaxation is decreased in mesenteric arteries from 12 month-old mRen2 female rats. We hypothesized that aging-induced GPER downregulation is present in other rodent species and strains as well. Aortas from young (2-3 months) and aged (11-23 months) C57BL/6 and BALB/c mice were lysed and immunoblotted for GPER protein. In addition, mesenteric arteries from C57BL/6 mice were mounted on a wire myograph and assessed for vasorelaxation in response to estrogen receptor agonists. We found a significant downregulation of GPER in aging aortas (3.1 ± 0.6 vs. 0.63 ± 0.42, P=0.016, N=4 per group). Moreover, we found that vasodilation to the GPER agonist G-1 was significantly attenuated in aging resistance arteries (22 ± 7.5% vs 58 ± 6.5%, P=0.023, N=2-4). Interestingly, vasodilation to the nonselective receptor agonist estradiol was not altered by aging (24 ± 10% vs. 32 ± 2.0%, P=0.23, N=2-4). In light of our previous findings in the rat vasculature, our data indicate that aging-induced decreases in vascular GPER expression and function are conserved across vascular beds and rodent species. We propose that aging-induced GPER downregulation switches the vascular benefits of postmenopausal estrogen therapy from positive to negative. Our future goal is to determine whether therapies that target GPER improve cardiovascular outcomes to protect the aging female population.

The Data Acquisition System for the CDMS II dark matter experiment was designed and built when the experiment moved to its new underground installation at the Soudan Lab. The combination of remote operation and increased data load necessitated a completely new design. Elements of the original LabView system remained as stand-alone diagnostic programs, but the main data processing moved to a VME-based system with custom electronics for signal conditioning, trigger formation and buffering. The data rate was increased 100-fold and the automated cryogenic system was linked to the data acquisition. A modular server framework with associated user interfaces was implemented in Java to allow control and monitoring of the entire experiment remotely.

Background: Rheumatoid arthritis is a chronic progressive autoimmune disease affecting 1.5 million people in the United States. Uncontrolled disease can lead to irreversible joint damage, deformity...

Funded by the European Space Agency (ESA), the Ariane 5 Program has been developed under the responsibility of the CNES, the French Space Agency. Attitude Control of the Ariane 5 Mission is carried out by the SCA (Systeme Controle d'Attitude), developed by Astrium-Space Transportation (Astrium-ST). It is the blow down hydrazine system using gaseous nitrogen as pressurant. The ARIANE 5 -SCA Tank - Fig.1 - is manufactured from TiAl6V4-hemispheres essentially performed via Super Plastic Forming (SPF) process. The tank shell wall thickness achieved need not to be mechanically reworked except small domains prepared for welding. Vis-a-vis the common tank production which employs hemispheres resulted by mechanical milling out of forged shell, the SPF provides certainly saving on milling efforts as well as significant reduction on material wasting. Also shown that is for tank component submitted to space application, the SPF as production method commonly undergoes development and verification efforts in engineering elaborating focused on Test-shell, Qualification-shell and such which provides samples for material strength and technological verifications. Enclosing is also a brief overall display addressing successful use of SPF-shell in producing tanks for the space flight today. Up to now more then 100 Tanks have been manufactured by Astrium-ST and > 90 Tanks were successful used in Ariane 5 launchers with various missions.

The expected final reach of the Weakly Interacting Massive Particle (WIMP) search experiment CDMS-II by the end of 2007 is a WIMP-nucleon cross-section sensitivity of 2.1×10−44 cm2. To proceed further in our search, we have proposed the SuperCDMS Phase A project that would deploy 42 1-inch thick Ge detectors, at a site deeper than the location of CDMS II, and reach a desired sensitivity goal of 1.3×10−45 cm2. These cross-sections are of interest and are complementary to Supersymmetry searches at the Large Hadron Collider (LHC) and future linear colliders.

Mutations in the gene for polycystin 2 (Pkd2) lead to polycystic kidney disease, however the main cause of mortality in humans is cardiac related. We previously showed that 5 month old Pkd2+/- mice have altered calcium-contractile activity in cardiomyocytes, but have preserved cardiac function. Here, we examined 1 and 9 month old Pkd2+/- mice to determine if decreased amounts of functional polycystin 2 leads to impaired cardiac function with aging. We observed changes in calcium handling proteins in 1 month old Pkd2+/- mice, and these changes were exacerbated in 9 month old Pkd2+/- mice. Anatomically, the 9 month old Pkd2+/- mice had thinner left ventricular walls, consistent with dilated cardiomyopathy, and the left ventricular ejection fraction was decreased. Intriguingly, in response to acute isoproterenol stimulation to examine β-adrenergic responses, the 9 month old Pkd2+/- mice exhibited a stronger contractile response, which also coincided with preserved localization of the β2 adrenergic receptor. Importantly, the Pkd2+/- mice did not have any renal impairment. We conclude that the cardiac-related impact of decreased polycystin 2 progresses over time towards cardiac dysfunction and altered adrenergic signaling. These results provide further evidence that polycystin 2 provides a critical function in the heart, independent of renal involvement.

Purpose Age-related macular degeneration (AMD), the most common cause of incurable blindness in the western world, is characterized by the dysfunction and eventual death of choroidal endothelial (CECs), RPE, and photoreceptor cells. Stem cell-based treatment strategies designed to replace photoreceptor and RPE cells currently are a major scientific focus. However, the success of these approaches likely also will require replacement of the underlying, supportive choroidal vasculature. The purpose of this study was to generate stem cell-derived CECs to develop efficient differentiation and transplantation protocols. Methods Dermal fibroblasts from the Tie2-GFP mouse were isolated and reprogrammed into two independent induced pluripotent stem cell (iPSC) lines via viral transduction of the transcription factors Oct4, Sox2, Klf4, and c-Myc. Tie2-GFP iPSCs were differentiated into CECs using a coculture method with either the RF6A CEC line or primary mouse CECs. Induced pluripotent stem cell-derived CECs were characterized via RT-PCR and immunocytochemistry for EC- and CEC-specific markers. Results Induced pluripotent stem cells generated from mice expressing green fluorescent protein (GFP) under control of the endothelial Tie2 promoter display classic pluripotency markers and stem cell morphology. Induced pluripotent stem cell-derived CECs express carbonic anhydrase IV, eNOS, FOXA2, PLVAP, CD31, CD34, ICAM-1, Tie2, TTR, VE-cadherin, and vWF. Conclusions Induced pluripotent stem cell-derived CECs will be a valuable tool for modeling of choriocapillaris-specific insults in AMD and for use in future choroidal endothelial cell replacement approaches.

The damping behavior of several materials used in current thermal barrier coating systems are reported from room temperature up to 1200 °C. The effect of long-term aging at 1150 °C on the damping behavior of a PWA-1484 single crystal superalloy is presented as well as the effect of a platinum-modified diffusion nickel aluminide bond-coat and an electron-beam evaporated, yttria-stabilized zirconia (EB-YSZ) thermal barrier coating. The nickel aluminide bond coat is shown to enhance the damping of the superalloys at all temperatures while the EB-YSZ shows a damping peak at ∼ 220 °C. The damping behavior of two top-coat materials, tetragonal-prime YSZ and gadolinium zirconate is compared. The latter exhibits a damping peak at a higher temperature (∼ 350 °C) than zirconia giving it the potential of being used as a damping coating for compressor blade.

The published CDMS analyses have used the shape of the phonon signal rising edge to reject low-ionization-yield surface events which produce acoustic phonons more quickly than bulk events do. To achieve better WIMP sensitivity with future larger exposures, we are using a simplified model of phonon production and propagation to construct event position estimators that help us to find more efficient surface event rejection cuts. We describe this model and the resulting new cuts, and summarize the surface event leakage rates and the sensitivity figures of merit of the five surface event rejection methods developed in the second CDMS II Soudan run data analysis.

In the muon ( g - 2 ) experiment at Brookhaven National Laboratory, the spin precession frequency ω a is obtained from a standard χ 2 minimization fit applied to the time distribution of decay electrons. The unusually high accuracy ( ∼ 0.5 ppm ) of the experiment puts stringent requirements on the quality of the fit and the level of understanding of the statistical properties of the fitted parameters. We discuss the properties of the fits and their implications on the derived value for ω a , including estimates of the effect of an imperfect fit function, methods of including additional external information to reduce the error, the effects of splitting the data into many smaller subsets of data, applying different weighting methods to the data using energy information, and various tests of data suitability.

In the past year and a half, the Cryogenic Dark Matter Search (CDMS) collaboration has been active at the Soudan mine in installing a system for running ZIP detectors that will be used to search for dark matter in the form of Weakly Interacting Massive Particles. Presently, there is an operating cryogenic system, working electronics, a functional data acquisition and analysis system, passive shielding, an active muon veto, and 12 ZIP detectors. Six of the 12 ZIP detectors have been tested in situ and are fully operational with acceptable noise profiles. CDMS is in the process of commissioning the experiment and expects to be making a background measurement by the end of summer 2003.

The experimental method together with the analysis method and results of the data taken in 2000 and prospects of the muon anomalous magnetic and electric dipole moment experiments are presented here., 12 pages, 20 figures. Proceedings of my plenary talk at ICHEP02, Amsterdam, 31 July 2002

1. Thoracic insufficiency syndrome is the inability of the thorax to support normal respiration or lung growth. 2. The rare condition of fused ribs and congenital scoliosis may result in a three-dimensional thoracic deformity with adverse effects on thoracic growth and function with development of thoracic insufficiency syndrome. 3. The normal thorax is defined by two characteristics: normal, stable volume and the ability to change that volume. Volume depends on the width and depth of the rib cage, and the thoracic spine provides height. The ability to change volume, termed thoracic function, is provided by the diaphragm and the secondary muscles of respiration. 4. On radiographs, the loss of the vertical height of the lung of the concave, restricted hemithorax is defined by the percentage of space available for the lung. 5. Spine rotation causes a windswept thorax, with both restriction of the volume of the convex hemithorax and restriction of the motion of the involved ribs. 6. Constrictive three-dimensional deformity of the thorax may cause extrinsic, restrictive lung disease. 7. Progressive thoracic insufficiency syndrome is diagnosed on the basis of clinical signs of respiratory insufficiency, loss of chest wall mobility as demonstrated by the thumb excursion test, worsening indices of three-dimensional thoracic deformity on radiographs and computed tomography scans, or a relative decline in percent predicted vital capacity due to thoracic "failure to thrive," as demonstrated by pulmonary function tests. 8. Treatment of progressive thoracic insufficiency syndrome should provide an acute increase in the thoracic volume with stabilization of any flail chest-wall defects and maintain these improvements as the patient grows, without the need for spine fusion.