ABSTRACT
The Primary Immune Deficiency Treatment Consortium (PIDTC) is a network of 33 centers in North America that study the treatment of rare and severe primary immunodeficiency diseases. Current protocols address the natural history of patients treated for severe combined immunodeficiency (SCID), Wiskott-Aldrich syndrome, and chronic granulomatous disease through retrospective, prospective, and cross-sectional studies. The PIDTC additionally seeks to encourage training of junior investigators, establish partnerships with European and other International colleagues, work with patient advocacy groups to promote community awareness, and conduct pilot demonstration projects. Future goals include the conduct of prospective treatment studies to determine optimal therapies for primary immunodeficiency diseases. To date, the PIDTC has funded 2 pilot projects: newborn screening for SCID in Navajo Native Americans and B-cell reconstitution in patients with SCID after hematopoietic stem cell transplantation. Ten junior investigators have received grant awards. The PIDTC Annual Scientific Workshop has brought together consortium members, outside speakers, patient advocacy groups, and young investigators and trainees to report progress of the protocols and discuss common interests and goals, including new scientific developments and future directions of clinical research. Here we report the progress of the PIDTC to date, highlights of the first 2 PIDTC workshops, and consideration of future consortium objectives.
Subject(s)
Immunologic Deficiency Syndromes , Hematopoietic Stem Cell Transplantation , Humans , Immunologic Deficiency Syndromes/diagnosis , Immunologic Deficiency Syndromes/therapy , Infant, Newborn , Neonatal Screening , Pilot Projects , Societies, ScientificABSTRACT
The current best serum marker for pancreatic cancer, CA 19-9, detects a carbohydrate antigen on multiple protein carriers. Better knowledge of the protein carriers of the CA 19-9 antigen in various disease states may lead to improved diagnostic tests. To identify proteins that carry the CA 19-9 antigen, we immunoprecipitated the CA 19-9 antigen from pooled sera and identified the associated proteins using MS. Among the high-confidence identifications, we confirmed the presence of the CA 19-9 antigen on Apolipoprotein B-100 by antibody arrays and Western blot and on kininogen, ARVCF, and Apolipoprotein E by antibody arrays. We characterized the frequency and levels of the CA 19-9 antigen on the four proteins across various patient groups (pancreatic cancer, pancreatitis, and healthy controls) using antibody arrays. Nearly, 10-25% of the subjects showed elevations of the antigen on each protein, but the elevations were not associated with disease state or total CA 19-9 levels. These results contribute to our knowledge of the carrier proteins of an important functional glycan and the rate at which the glycan is displayed. This work also demonstrates a strategy for using the complementary methods of MS and antibody microarrays to identify protein carriers of glycans and assess the diagnostic value of measuring glycans on individual proteins.
Subject(s)
CA-19-9 Antigen/blood , Carrier Proteins/immunology , Mucins/isolation & purification , Protein Array Analysis/methods , Biomarkers/chemistry , CA-19-9 Antigen/chemistry , Carrier Proteins/chemistry , Case-Control Studies , Humans , Immunoprecipitation , Mass Spectrometry/methods , Mucins/blood , Mucins/chemistry , Mucins/immunology , Pancreatic Neoplasms/blood , Pancreatic Neoplasms/immunology , Pancreatitis/blood , Pancreatitis/immunology , Proteomics/methods , Sensitivity and SpecificityABSTRACT
Clostridium sordellii is a toxin-producing anaerobic bacillus that causes severe infections in humans and livestock. C. sordellii infections can be accompanied by a highly lethal toxic shock syndrome (TSS). Lethal toxin (TcsL) is an important mediator of TSS. We recently obtained a clinical strain of C. sordellii (DA-108) lacking the TcsL-encoding tcsL gene, which was not fatal in rodent models of infection, in contrast to a tcsL(+) reference strain (ATCC9714). Protein preparations derived from cell-free, stationary phase cultures obtained from ATCC9714 were lethal when injected into mice, while those obtained from DA-108 were not, a difference that was attributed to the unique presence of TcsL in the ATCC9714-derived proteins. We questioned whether there were other major differences between the extracellular proteomes of these two strains, apart from TcsL. Two-dimensional gel electrophoresis was conducted using crude cell-free supernatants from these strains and 14 differentially expressed proteins were subjected to mass spectrometric analysis. Nine of these 14 proteins were more highly expressed by DA-108 and 5 by ATCC9714. Twelve of the 14 proteins isolated from the 2-D gels were putatively identified by mass spectrometry. Several of these proteins were identical, possibly reflecting enzymatic cleavage, degradation, and/or post-translational modifications. Excluding identical sequences, only 5 unique proteins were identified. Four proteins (ferredoxin-nitrite reductase; formate acetyltransferase; Translation Elongation Factor G; and purine nucleoside phosphorylase) were over-expressed by DA-108 and 1 (N-acetylmuramoyl-l-alanine amidase) by ATCC9714. These results support the concept that TcsL is the major determinant of C. sordellii TSS during infection.
Subject(s)
Bacterial Proteins/analysis , Bacterial Proteins/metabolism , Clostridium sordellii/chemistry , Proteome/analysis , Virulence Factors/analysis , Bacterial Toxins/genetics , Clostridium sordellii/pathogenicity , Electrophoresis, Gel, Two-Dimensional , Mass SpectrometryABSTRACT
Structural and cellular attachment analysis identified overall bent helical regions of adhesive peptides identified within mussel adhesive protein (MAP) capable of also attaching cells. DOPA (L-DOPA, 3,4-dihydroxyphenylalanine) is frequently identified and credited for the attachment ability of several marine proteins [Olivieri, MP, et al. (2002), Biofouling18, 149-159]. Newly designed cyclic peptides (DOPA-G-G-C-G-K-A-K-G-C [cyc-DOPA] & Y-G-G-C-G-K-A-K-G-C [cyc-Y]) derived from structurally conserved regions of several MAP peptides were examined to assist in the understanding of both surface and cellular attachment. Solution-state proton nuclear magnetic resonance (NMR) spectroscopy coupled with molecular modeling and dynamics revealed minimal differences in the structures of the proposed cellular attachment domain within these two peptides. Multiple attenuated internal reflection infrared (MAIR-IR) spectroscopy, ellipsometry and advancing contact angle analyses showed that formation of thin films by these peptides was L-DOPA and pH dependent. When compared to control surfaces, undifferentiated leukocyte cells (MOLT-4) significantly attached and spread onto films created from the cyc-DOPA. The culmination of these structural, biophysical and cellular attachment techniques reveal a conformation of cyc-DOPA that is capable of both adsorbing to surfaces and then attaching cells that spread. This work supports the sequence, K-A-K, as the cellular attachment domain, especially when held in a reliable structural conformation.,.
ABSTRACT
OBJECTIVES: To study 11 patients with melanoma-associated retinopathy (MAR) to clarify the reliability of various methods of diagnostic testing, to determine the underlying antigenic retinal proteins, and to study the clinical histories and types of associated melanomas. METHODS: Clinical data were obtained from patients with melanoma who developed marked visual problems. Testing included electroretinography, kinetic visual fields, comparative studies of Western blots, and indirect immunohistologic examination to detect antiretinal antibodies, as well as proteomic studies to identify underlying antigenic retinal proteins. RESULTS: Patients with MAR typically have rapid onset of photopsias, scotomata, and loss of central or paracentral vision. Ophthalmoscopy seldom shows significant changes early, but electroretinograms are abnormal. Results of Western blots and immunohistologic examination can show antiretinal antibodies but not always. Most patients (9 of 11) had a strong family history of autoimmune disorders. Any type of melanoma (cutaneous, choroidal, ciliary body, or choroidal nevi) may be associated with this paraneoplastic autoimmune reactivity. MAR may precede or follow the diagnosis of melanoma. Patients with MAR have the same antigenic retinal proteins that have been associated with cancer-associated retinopathy. In addition, 2 new antigenic retinal proteins, aldolase A and aldolase C, were found. CONCLUSIONS: There was a high prevalence of positive family histories of autoimmune disease in patients with MAR. To confirm the disorder, multiple clinical and serum diagnostic techniques (Western blot or indirect immunohistologic examination) are needed. Two newly observed antigenic retinal proteins, aldolase A and aldolase C, are associated with MAR.
Subject(s)
Autoantibodies/blood , Autoantigens/analysis , Autoimmune Diseases/immunology , Melanoma/immunology , Paraneoplastic Syndromes/immunology , Retinal Diseases/immunology , Adult , Aged , Aged, 80 and over , Autoimmune Diseases/etiology , Blotting, Western , Electrophoresis, Polyacrylamide Gel , Electroretinography , Female , Fluorescent Antibody Technique, Indirect , Fructose-Bisphosphate Aldolase/analysis , Humans , Male , Melanoma/pathology , Middle Aged , Paraneoplastic Syndromes/etiology , Retinal Diseases/etiology , Scotoma/etiology , Skin Neoplasms/immunology , Skin Neoplasms/pathology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Uveal Neoplasms/immunology , Uveal Neoplasms/pathology , Visual FieldsSubject(s)
Health Care Reform , Politics , Chronic Disease/therapy , Disease Management , Fees, Pharmaceutical/legislation & jurisprudence , Humans , Medicaid/legislation & jurisprudence , Medicare/legislation & jurisprudence , Nursing , United States , Universal Health Insurance/legislation & jurisprudence , WorkforceABSTRACT
OBJECTIVES: Personal emergency response systems (PERSs) are reported to reduce anxiety and health care use and may assist in planning the disposition of older patients discharged from the emergency department (ED) to home. This study measured the impact of a PERS on anxiety, fear of falling, and subsequent health care use among older ED patients. METHODS: This study was a randomized controlled trial comparing PERS use with standard ED discharge planning in subjects 70 years of age or older discharged home after a fall. Outcome assessors were blinded to the study objectives. Anxiety and fear of falling were measured at baseline and 30 days using the Hospital Anxiety and Depression Scale anxiety subscale (HADS-A) and modified Falls Efficacy Scale (mFES). Return to the ED, hospitalization, and length of stay were recorded after 30 and 60 days. RESULTS: Eighty-six subjects were randomized and completed follow up (43 per group). There was no important difference in mean reduction in anxiety (mean change treatment - control, +0.35; 95% confidence interval [CI] = -1.5 to 0.76; p = 0.55) or fear of falling (mean change, +4.5; 95% CI = -6.7 to 15.7; p = 0.70). Return visits to the ED occurred in eight of 43 patients in both the control and treatment groups (risk difference, 0.0%; 95% CI = -16% to 16%). Hospitalization occurred in six of 43 in the control group versus three of 43 in the treatment group (risk difference treatment - control = -7.0%; 95% CI = -19.8% to 5.9%). CONCLUSIONS: In contrast to previous studies, there was no evidence that a PERS reduced anxiety, fear of falling, or return to the ED among older persons discharged from the ED.
Subject(s)
Accident Prevention/instrumentation , Accidental Falls , Anxiety/prevention & control , Anxiety/psychology , Emergency Medical Service Communication Systems , Emergency Service, Hospital/statistics & numerical data , Fear/psychology , Aged , Chi-Square Distribution , Female , Humans , Length of Stay/statistics & numerical data , Male , Statistics, NonparametricABSTRACT
NURSE MANAGERS are responsible for the satisfaction of both patients and staff. At first glance, this statement may seem both reasonable and attainable yet, for many years, achieving staff satisfaction has been elusive.
ABSTRACT
The zymogen granule (ZG) is the specialized organelle in pancreatic acinar cells for digestive enzyme storage and regulated secretion and has been a model for studying secretory granule functions. In an initial effort to comprehensively understand the functions of this organelle, we conducted a proteomic study to identify proteins from highly purified ZG membranes. By combining two-dimensional gel electrophoresis and two-dimensional LC with tandem mass spectrometry, 101 proteins were identified from purified ZG membranes including 28 known ZG proteins and 73 previously unknown proteins, including SNAP29, Rab27B, Rab11A, Rab6, Rap1, and myosin Vc. Moreover several hypothetical proteins were identified that represent potential novel proteins. The ZG localization of nine of these proteins was further confirmed by immunocytochemistry. To distinguish intrinsic membrane proteins from soluble and peripheral membrane proteins, a quantitative proteomic strategy was used to measure the enrichment of intrinsic membrane proteins through the purification process. The iTRAQ ratios correlated well with known or Transmembrane Hidden Markov Model-predicted soluble or membrane proteins. By combining subcellular fractionation with high resolution separation and comprehensive identification of proteins, we have begun to elucidate zymogen granule functions through proteomic and subsequent functional analysis of its membrane components.
Subject(s)
Enzyme Precursors , Intracellular Membranes/chemistry , Intracellular Membranes/metabolism , Membrane Proteins/analysis , Pancreas/cytology , Proteomics , Secretory Vesicles/metabolism , Animals , Membrane Proteins/isolation & purification , Protein Transport , Rats , Secretory Vesicles/chemistrySubject(s)
Hypertension/diagnosis , Hypertension/therapy , Practice Guidelines as Topic , Severity of Illness Index , Antihypertensive Agents/therapeutic use , Humans , Hypertension/complications , Hypertension/epidemiology , Life Style , Nurse's Role , Patient Selection , Primary Prevention/methods , United States/epidemiologyABSTRACT
BACKGROUND: Melasma is an acquired hypermelanosis that is often recalcitrant to treatment with hypopigmenting agents. OBJECTIVE: To assess the efficacy of 4% hydroquinone cream vs 4% hydroquinone cream combined with glycolic acid peels as treatment for melasma. METHODS: Twenty-one Hispanic women with bilateral epidermal and mixed melasma were enrolled in a split-faced prospective trial lasting 8 weeks. Patients underwent 20% to 30% glycolic acid peels every 2 weeks to one side of the face only in addition to twice-daily full-face application of 4% hydroquinone cream and sun protective factor 25 UV-B sunscreen each morning. Pigmentation was measured objectively using a mexameter and the Melasma Area and Severity Index and subjectively using a linear analog scale and physician and patient global evaluation. RESULTS: Hydroquinone treatment alone and treatment with the combination of hydroquinone and glycolic acid had a significant effect in reducing skin pigmentation compared with baseline (P<.001). However, no significant difference was found using combination therapy compared with hydroquinone alone (P =.75). CONCLUSIONS: Use of 4% hydroquinone and a daily sunscreen is effective in the treatment of melasma; however, the addition of 4 glycolic acid peels did not enhance the hypopigmenting effect of hydroquinone treatment alone.
Subject(s)
Chemexfoliation/methods , Facial Dermatoses/drug therapy , Glycolates/therapeutic use , Keratolytic Agents/therapeutic use , Melanosis/drug therapy , Adolescent , Adult , Aged , Esthetics , Facial Dermatoses/diagnosis , Female , Follow-Up Studies , Humans , Melanosis/diagnosis , Middle Aged , Patient Satisfaction , Prospective Studies , Statistics, Nonparametric , Treatment OutcomeABSTRACT
Proximal femoral valgus osteotomy for Legg-Calvé-Perthes disease was evaluated at an average 5 years postoperatively in 31 consecutive patients. The indications for osteotomy were hinge abduction and pain. The Iowa hip scores at follow-up for 21 patients averaged 93 points. Combined clinical and radiographic review for these patients yielded 6 (29%) excellent, 7 (33%) good, 5 (24%) fair, and 3 (14%) poor results. The 10 remaining patients had good pain relief and were satisfied but were unable to return for hip scale evaluation.