Health-related quality of life of adult and adolescent patients living with alopecia areata in Australia.

INTRODUCTION: To understand the experiences of adolescent and adult patients living with alopecia areata (AA) in Australia regarding symptom severity and the impact on psychosocial well-being and work/classroom productivity. MATERIALS AND METHODS: A cross-sectional online patient survey among adolescent and adult patients diagnosed with AA was recruited via the Australia Alopecia Areata Foundation. Patient-reported outcomes were also assessed. RESULTS: A total of 337 patients (49 adolescents; 288 adults), with a mean ± standard deviation age of 14.7 ± 1.55 and 38.9 ± 13.31 years for adolescents and adults, respectively, were included. In the group with extensive hair loss (Scalp Hair Assessment Patient-Reported Outcome, categories 3 + 4, n = 172), we observed higher emotional symptom and activity limitation scores (Alopecia Areata Patient Priority Outcomes, emotional symptoms: adults 2.5 ± 1.03, adolescents 2.2 ± 1.15; activity limitations: adults 1.4 ± 1.15, adolescents 1.2 ± 0.99). Additionally, in adults, the Alopecia Areata Symptom Impact Scale global score was 4.0 ± 2.10 (symptoms subscale score 4.1 ± 1.91; interference subscale scores 3.8 ± 2.73). Hospital Anxiety and Depression Scale scores were high across participants, irrespective of hair loss extent (adults: anxiety 9.2 ± 3.85, depression 6.6 ± 3.95; adolescents: anxiety 9.7 ± 4.65, depression 5.2 ± 3.59). Work and classroom productivity were substantially impaired due to AA, with 70.5% of adults and 57.1% of adolescents reporting activity impairment, and overall work/classroom impairment reported at 39.2% and 44.9%, respectively. CONCLUSIONS: AA impacts the physical, emotional and psychosocial well-being of both adult and adolescent patients. More extensive hair loss more profoundly impacts those living with AA. Patients may benefit from patient-centred care approaches addressing the impact of hair loss on mental and emotional well-being, daily activities and work productivity.

Comparison of pelvic incidence measurement using lateral x-ray, standard ct versus ct with 3d reconstruction.

PURPOSE: Pelvic incidence (PI) is a position independent parameter used to quantify spinopelvic sagittal balance. PI is generally measured on lateral radiographs, but more recent studies have suggested better accuracy with standard CT scans versus three-dimensional (3D) CT scans. This study compares PI obtained from lateral XR, standard CT scan and CT scan with 3D reconstruction. METHODS: A total of 77 subjects with lateral XRs of the pelvis or lumbosacral spine and CT scans of the pelvis were randomly selected. Pelvic incidence on lateral XRs, standard CT scans and CT scans utilizing multiplanar reconstruction were measured and compared using intraclass correlation coefficients (ICC). PI was also measured on serial images in 28 individuals using the same imaging modality within 3 years and evaluated using ICC. RESULTS: Mean ± SD of PI measurements on XR, standard CT and CT with 3D reconstruction were 56° ± 13°, 53° ± 12° and 53° ± 12°, respectively, demonstrating a small but significant elevation of PI measurement on XR (P < 0.001). ICC values demonstrated a higher correlation between standard CT and 3D CT (ICC 0.986), compared to XR and standard CT (ICC 0.934) and XR and 3D CT (ICC 0.937). PI measurements on repeated imaging of the same individual also demonstrated that both CT methods produced more consistent measurements (ICC 0.986 for standard CT, 0.981 for 3D CT, 0.935 for XR). CONCLUSION: Although standard XR does provide a high level of reliability, it appears to slightly overestimate PI. CT scans do provide increased reliability, with no additional benefit of 3D reconstructions over standard CT.

The distribution of the therapeutic monoclonal antibodies cetuximab and trastuzumab within solid tumors.

BACKGROUND: Poor distribution of some anticancer drugs in solid tumors may limit their anti-tumor activity. METHODS: Here we used immunohistochemistry to quantify the distribution of the therapeutic monoclonal antibodies cetuximab and trastuzumab in relation to blood vessels and to regions of hypoxia in human tumor xenografts. The antibodies were injected into mice implanted with human epidermoid carcinoma A431 or human breast carcinoma MDA-MB-231 transfected with ERBB2 (231-H2N) that express high levels of ErbB1 and ErbB2 respectively, or wild-type MDA-MB-231, which expresses intermediate levels of ErbB1 and low levels of ErbB2. RESULTS: The distribution of cetuximab in A431 xenografts and trastuzumab in 231-H2N xenografts was time and dose dependent. At early intervals after injection of 1 mg cetuximab into A431 xenografts, the concentration of cetuximab decreased with increasing distance from blood vessels, but became more uniformly distributed at later times; there remained however limited distribution and binding in hypoxic regions of tumors. Injection of lower doses of cetuximab led to heterogeneous distributions. Similar results were observed with trastuzumab in 231-H2N xenografts. In MDA-MB-231 xenografts, which express lower levels of ErbB1, homogeneity of distribution of cetuximab was achieved more rapidly. CONCLUSIONS: Cetuximab and trastuzumab distribute slowly, but at higher doses achieve a relatively uniform distribution after about 24 hours, most likely due to their long half-lives in the circulation. There remains poor distribution within hypoxic regions of tumors.

Streptococcus Dysgalactiae Subspecies Equisimilis Endogenous Endophthalmitis Associated with Aortic Valve Abscess.

Purpose: To describe a case of endogenous endophthalmitis from Streptococcus dysgalactiae subspecies equisimilis in the setting of an aortic valve abscess Methods: Retrospective case report. Results: A 72-year-old white male presented with fevers, encephalopathy, and decreased vision in his left eye. His visual acuity was 20/20 in his right eye and finger counting in the left eye. Workup revealed an aortic valve abscess. Examination of his left eye revealed dense anterior chamber fibrin and no view of the retina. B-scan ultrasonography revealed loculated hyperechoic areas consistent with vitreous inflammation. A vitreous tap and injections with vancomycin and ceftazidime were performed. Visual acuity worsened to no-light perception 5 days later. Vitreous and blood cultures grew Streptococcus dysgalactiae subspecies equisimilis. He received another intravitreal injection of vancomycin with no improvement. Conclusions: Streptococcus dysgalactiae subspecies equisimilis is an emerging pathogen and may cause severe intraocular infections with a poor visual outcome.

Limited penetration of anticancer drugs through tumor tissue: a potential cause of resistance of solid tumors to chemotherapy.

PURPOSE: Potential causes of drug resistance in solid tumors include genetically determined factors expressed in individual cells and those related to the solid tumor environment. Important among the latter is the requirement for drugs to penetrate into tumor tissue and to achieve a lethal concentration in all of the tumor cells. The present study was designed to characterize further the multicellular layer (MCL) method for studying drug penetration through tumor tissue and to provide information about tissue penetration for drugs used commonly in the treatment of human cancer. EXPERIMENTAL DESIGN: EMT-6 mouse mammary and MGH-U1 human bladder cancer cells were grown on collagen-coated semiporous Teflon membranes to form MCLs approximately 200 microm thick. The properties of MCLs were compared with those of tumors grown in mice from the same cells. The penetration of drugs through the MCL was evaluated by using radiolabeled drugs or analytical methods. RESULTS: The MCL developed an extracellular matrix containing both laminin and collagen, although there were some differences in expression of extracellular matrix proteins. Electron microscopy showed rare desmosomes in both MCL and tumors. The penetration of cisplatin, etoposide, gemcitabine, paclitaxel, and vinblastine through tissue in the MCL was slow compared with penetration through the Teflon support membrane alone. CONCLUSIONS: Our results suggest limited ability of anticancer drugs to reach tumor cells that are distant from blood vessels. The limited penetration of anticancer drugs through tumor tissue may be an important cause of clinical resistance of solid tumors to chemotherapy.

MRI monitoring of intratumoral drug delivery and prediction of the therapeutic effect with a multifunctional thermosensitive liposome.

Non-invasive in vivo imaging of drug distribution enables real-time monitoring and prediction of therapeutic responses to treatment. We have developed a thermosensitive liposomal formulation (HaT: Hyperthermia-activated-cytoToxic) consisting of DPPC and Brij78, a formulation that enhanced drug delivery compared to the lyso-lipid temperature sensitive liposomes (LTSL). Here we report the development of a multifunctional HaT liposome co-encapsulating Gd-DTPA (an MRI probe) and doxorubicin (DOX), which simultaneously releases and reports on drug delivery in a locally heated tumor. The temperature-dependent release profiles of DOX from HaT were closely related to the change in the MR T(1) relaxation time, in which DOX was 100% released at 40-42 °C in 3 min, accompanied by a 60% reduction in T(1). By T(1) relaxometry analysis, no Gd-DTPA leakage was detected in 30 min at 30-37 °C. In the in vivo study, DOX uptake in the tumor was quantitatively correlated with T(1) response (R(2) = 0.98) and the patterns of the T(1) image and the intratumoral DOX uptake were matched, in which both signals were predominantly detected in the highly perfused tumor periphery. Finally, the extent of T(1) relaxation enhancement in the heated tumor successfully predicted the antitumor efficacy in a standard pharmacological response model (R(2) = 0.98).

Interleukin-21 is critically required in autoimmune and allogeneic responses to islet tissue in murine models.

OBJECTIVE: Type 1 diabetes is an incurable chronic autoimmune disease. Although transplantation of pancreatic islets may serve as a surrogate source of insulin, recipients are subjected to a life of immunosuppression. Interleukin (IL)-21 is necessary for type 1 diabetes in NOD mice. We examined the efficacy of an IL-21-targeted therapy on prevention of diabetes in NOD mice, in combination with syngeneic islet transplantation. In addition, we assessed the role of IL-21 responsiveness in islet allograft rejection in mouse animal models. RESEARCH DESIGN AND METHODS: NOD mice were treated with IL-21R/Fc, an IL-21-neutralizing chimeric protein. This procedure was combined with syngeneic islet transplantation to treat diabetic NOD mice. Survival of allogeneic islet grafts in IL-21R-deficient mice was also assessed. RESULTS: Evidence is provided that IL-21 is continually required by the autoimmune infiltrate, such that insulitis was reduced and reversed and diabetes inhibited by neutralization of IL-21 at a late preclinical stage. Recovery from autoimmune diabetes was achieved by combining neutralization of IL-21 with islet transplantation. Furthermore, IL-21-responsiveness by CD8+ T-cells was sufficient to mediate islet allograft rejection. CONCLUSIONS: Neutralization of IL-21 in NOD mice can inhibit diabetes, and when paired with islet transplantation, this therapeutic approach restored normoglycemia. The influence of IL-21 on a graft-mounted immune response was robust, since the absence of IL-21 signaling prevented islet allograft rejection. These findings suggest that therapeutic manipulation of IL-21 may serve as a suitable treatment for patients with type 1 diabetes.

Expression, purification and characterization of recombinant interleukin-21.

Interleukin-21 (IL-21) is a key regulator of the immune system. However, studies of this cytokine have so far been hampered by the limited availability of recombinant protein preparations. Here we describe a method based on refolding of inclusion bodies expressed in E. coli by rapid dilution. The method was applied to human and murine IL-21 proteins, which were further purified by affinity chromatography and gel-filtration. The proteins are pure and highly active as determined by endotoxin and cell proliferation assays. The availability of milligram quantities of protein enabled us to generate monoclonal antibody fragments against the cytokine and will aid in further structural, biochemical and physiological analyses.

Detection of growth hormone doping by gene expression profiling of peripheral blood.

CONTEXT: GH abuse is a significant problem in many sports, and there is currently no robust test that allows detection of doping beyond a short window after administration. OBJECTIVE: Our objective was to evaluate gene expression profiling in peripheral blood leukocytes in-vivo as a test for GH doping in humans. DESIGN: Seven men and thirteen women were administered GH, 2 mg/d sc for 8 wk. Blood was collected at baseline and at 8 wk. RNA was extracted from the white cell fraction. Microarray analysis was undertaken using Agilent 44K G4112F arrays using a two-color design. Quantitative RT-PCR using TaqMan gene expression assays was performed for validation of selected differentially expressed genes. RESULTS: GH induced an approximately 2-fold increase in circulating IGF-I that was maintained throughout the 8 wk of the study. GH induced significant changes in gene expression with 353 in women and 41 in men detected with a false discovery rate of less than 5%. None of the differentially expressed genes were common between men and women. The maximal changes were a doubling for up-regulated or halving for down-regulated genes, similar in magnitude to the variation between individuals. Quantitative RT-PCR for seven target genes showed good concordance between microarray and quantitative PCR data in women but not in men. CONCLUSION: Gene expression analysis of peripheral blood leukocytes is unlikely to be a viable approach for the detection of GH doping.

Selection of human antibody fragments by phage display.

Here, we describe a protocol for the selection of human antibody fragments using repertoires displayed on filamentous bacteriophage. Antigen-specific clones are enriched by binding to immobilized antigen, followed by elution and repropagation of phage. After multiple rounds of binding selection, specific clones are identified by ELISA. This article provides an overview of phage display and antibody technology, as well as detailed protocols for the immobilization of antigen, the selection of repertoires on purified or complex antigens and the identification of binders.