Esophageal distensibility measurement: impact on clinical management and procedure length.

Luminal distensibility measurement has demonstrated relevance to various disease processes, though its effects on clinical decision-making have been less well understood. This study aims to characterize the clinical impact of impedance planimetry measurement as well as the learning curve associated with its use in the esophagus. A single provider performed distensibility measurement in conjunction with upper endoscopy for a variety of clinical indications with the functional lumen imaging probe (FLIP) over a period of 21 months. Procedural data were prospectively collected and, along with medical records, retrospectively reviewed. Seventy-three procedures (70 patients) underwent esophageal distensibility measurement over the timeline of this study. The most common procedural indications were known or suspected achalasia (32.9%), dysphagia with connective tissue disease (13.7%), eosinophilic esophagitis (12.3%), and dysphagia with prior fundoplication (9.6%). FLIP results independently led to a change in management in 29 (39.7%) cases and supported a change in management in an additional 15 (20.5%) cases. The most common change in management was a new or amended therapeutic procedure (79.5%). Procedural time added by distensibility measurement was greater among earlier cases than among later cases. The median time added overall was 5 minutes and 46 seconds. Procedural time added varied significantly by procedural indication, but changes in management did not. Distensibility measurement added meaningful diagnostic information that impacted therapeutic decision-making in the majority of cases in which it was performed. Procedural time added by this modality is typically modest and decreases with experience.

Impacts of trace mineral source and ancillary drench on steer performance during a 60-day backgrounding phase.

Nutritional approaches to optimize cattle health and performance during the receiving period are warranted. This experiment evaluated the impacts of supplementing organic complexed Cu, Co, Mn, and Zn on productive and health responses of high-risk beef cattle during a 60-day backgrounding phase. Crossbred steers (120) were purchased at auction and transported to the experimental facility, where BW was recorded (day-1; initial shrunk BW = 227.7 ± 1.3 kg). On day 0, steers were ranked by BW and allocated to one of eight groups and housed in drylot pens equipped with GrowSafe automated feeding systems (Model 8000; two bunks/pen). Groups were randomly assigned to receive a total mixed ration containing: (1) sulfate sources of Cu, Co, Mn, and Zn (INR; n = 40); (2) organic complexed sources of the same minerals (AAC; Zinpro Availa 4 based on a metal:amino acid complex ratio of 1:1 for Zn, Cu, and Mn in addition to cobalt glucoheptonate; Zinpro Corp., Eden Prairie, MN; n = 40); or (3) AAC and an organic complexed trace mineral drench (APF; 30 mL/hd; Zinpro ProFusion, Zinpro Corp.) on day 0 and with morbidity treatment (n = 40). Diets provided the same daily amount of all nutrients and minerals based on 7 g/steer daily of Zinpro Availa 4. Steers were assessed for bovine respiratory disease (BRD) signs daily. Liver biopsies were performed on days 0, 28 and 60. Blood samples were collected on days 0, 2, 6, 10, 13, 21, 28 and 45. No treatment differences were detected (P ≥ 0.23) for feed intake, final BW, average daily gain, or BRD incidence. Mean liver Co concentrations were greater (P = 0.02) in AAC and APF compared to INR steers. Mean liver Cu was greater (P = 0.02) in APF compared to AAC steers. Liver Zn tended to be greater (P = 0.10) on day 28 but less (P = 0.05) on day 60 for INR compared to AAC and APF steers. Plasma cortisol was lowest (P = 0.05) for AAC steers on day 6, whereas AAC steers tended to have greater (P = 0.09) plasma cortisol on day 13 compared with APF. Plasma haptoglobin tended to be greater (P ≤ 0.10) for INR steers on days 28 and 45 compared to AAC and APF. While supplementing cattle with AAC or INR results in similar animal performance and clinical disease, AAC and APF reduce stress and acute phase protein responses.

Comparison of growth requirements of two human intratumoral colon carcinoma cell lines in monolayer and soft agarose.

The human colon, intratumoral subpopulations HCT 116 and HCT 116a were established in chemically defined medium supplemented with transferrin, insulin, epidermal growth factor (EGF), triiodothyronine, hydrocortisone, and sodium selenite. The responsiveness of the adapted cell lines to these growth factors was compared in anchorage-dependent and -independent assays. HCT 116 cells maintained in serum-free conditions were further adapted to growth factor deprivation, and the effects of these polypeptides were determined in anchorage-independent assays. In monolayer, HCT 116 cells adapted to grow in serum-free medium responded to transferrin but not to EGF or insulin. Similarly adapted HCT 116a cells were, however, insensitive to transferrin addition but manifested a 300 and 500% increase in growth rates with EGF and insulin, respectively. Optimal growth of HCT 116 cells was seen in the presence of insulin and transferrin, while maximum proliferation of HCT 116a cells depended on combined insulin, transferrin, and EGF. In soft agarose, both HCT 116 and HCT 116a subpopulations showed a stringent requirement for transferrin. No combination of growth factors without transferrin supported colony formation. These data suggest that (a) these colon tumor subpopulations may be subject to separate growth controls, and (b) there may be an important role for transferrin in anchorage-independent growth and possibly in the maintenance of malignant characteristics.

Characterization of the inhibitory effects of transforming growth factor-beta on a human colon carcinoma cell line.

The effects of transforming growth factor-beta (TGF beta) on a human colon carcinoma cell line (MOSER) were investigated. TGF beta, at low concentrations (between 0.1 and 1.0 ng/ml), inhibited the proliferation of MOSER cells both in monolayer culture and soft agarose, in a dose-dependent manner. MOSER cells adapted to growth in chemically defined serum-free medium were more sensitive to the inhibitory effects of TGF beta than cells maintained in serum-supplemented medium. Morphological changes in MOSER cells, observed with TGF beta, were similar to those seen with the chemical differentiation agent N,N-dimethylformamide. Also in similarity to the effects of N,N-dimethylformamide, TGF beta induced a time- and concentration-dependent increase in soluble extracellular fibronectin. Binding studies with [125I]TGF beta revealed a relatively low number of binding sites on MOSER cells (13%) compared with mouse embryo fibroblastic (AKR-2B) cells. Thus far, other colon carcinoma cell lines, some displaying TGF beta receptors, have been reported to be unresponsive to TGF beta. This study is therefore the first to demonstrate a TGF beta-responsive colon carcinoma cell line.

Sensing textile seam-line for wearable multimodal physiological monitoring.

This paper investigates a novel multimodal sensing method by forming seam-lines of conductive textile fibers into commercially available fabrics. The proposed ultra-low cost micro-electro-mechanical sensor would provide, wearable, flexible, textile based biopotential signal recording, wetness detection and tactile sensing simultaneously. Three types of fibers are evaluated for their array-based sensing capability, including a 3D printed conductive fiber, a multiwall carbon nanotube based fiber, and a commercially available stainless steel conductive thread. The sensors were shown to have a correlation between capacitance and pressure; impedance and wetness; and recorded potential and ECG waveforms.

Craniofacial malformations and the orthodontist.

This review article presents an overview of craniofacial malformations and the role of the orthodontist in their management. The first part of this article focuses on cleft lip and palate, followed by more complex deformities including craniosynostosis and craniofacial microsomia. The main features of these anomalies are discussed as well as the clinical problems seen in this group of patients. The emphasis is on the role of the orthodontist in the multi-disciplinary management of these cases.

Different epidermal growth factor growth responses and receptor levels in human colon carcinoma cell lines.

The growth response to epidermal growth factor (EGF) and the numbers and types of EGF receptors were studied in three human colon tumor cell lines from each of two groups of cell lines that differ markedly in their growth properties and extent of differentiation. Aggressively growing and poorly differentiated colon cells (group I) did not respond to EGF alone, while less aggressively growing and more differentiated cells (group III) responded with increased growth when EGF was added to their chemically defined, serum-free medium. The average number of EGF receptors (EGF-R) measured at the surface of group III cell lines by radioligand binding assays, was eight-fold higher than that measured for group I cell lines. These observations provide evidence for possible autocrine mechanisms that maintain available EGF-R levels in more differentiated group III colon tumor cells and down-regulate EGF-R levels in group I colon tumor cells.

Comparison of the antiproliferative effects of transforming growth factor-beta, N,N-dimethylformamide and retinoic acid on a human colon carcinoma cell line.

In this study we have compared the anti-proliferative effects of transforming growth factor-beta (TGF-beta), N,N-dimethylformamide (DMF) and retinoic acid (RA) on a moderately-differentiated colon carcinoma cell line (JVC). TGF-beta, DMF and RA inhibited the anchorage-independent growth and induced morphological changes in JVC cells. EC50 values of 40 pM TGF-beta, 0.5% DMF and 5 nM RA were obtained for growth inhibition. In addition all three agents enhanced cellular fibronectin levels in a time- and dose-dependent manner. Inhibition of cell proliferation as well as fibronectin induction by all three agents were reversible. Combinations of any two agents at suboptimal doses, added simultaneously to JVC cells gave additive inhibitory response on growth. These data indicate that the effects of TGF-beta in this colon carcinoma cell line are similar to those of the two differentiation promoting agents DMF and RA.

Immunotherapy for recurrent colorectal cancers with human monoclonal antibody SK-1.

BACKGROUND: The human monoclonal antibody SK-1 recognizes a glycoprotein expressed on the majority of colon cancer tissues. In the current study, we evaluated the safety, toxicity and preliminary efficacy of escalating dosages of SK-1 in patients with advanced colon cancer. PATIENTS AND METHODS: SK-1 was administered intravenously at 2, 4 or 10 mg three times to three groups of patients with recurrent colon cancer. The clinical outcome and the induction of serum anti-idiotypic antibody (Ab2) were assessed periodically. RESULTS: The mean rate of serum CEA level increase declined significantly during the eight weeks following the treatment. In four patients, serum titer of anti-idiotypic IgG antibodies to SK-1 (Ab2) continued to increase following the treatment. CONCLUSION: HuMAb SK-1 was well-tolerated and can be safely administered. It was suggested that SK-1 natural antibody not only possessed direct cytostatic activity against colon carcinoma, but may also have induced carcinoma-related, anti-idiotypic antibody responses.

Co-expression of tumor antigens and their modulation by pleiotrophic modifiers enhance targeting of human monoclonal antibodies to pancreatic carcinoma.

Cocktails of human monoclonal antibodies (HuMAbs) have been used to increase the likelihood of identifying heterogeneous cancer cells. We utilized 3 HuMAbs termed SK-1, GM4, and GMA1, which recognized a 42-46 kDa two chain structure, a 57 kDa antigen, and the ganglioside GD3, respectively. An estimated two dozen cell lines were tested for the coexpression of these antigens and this was found to be present only on pancreatic carcinoma cell line, PANC-1; A 24 hr treatment of PANC-1 cells with interferon gamma (IFN gamma; 100 units), interferon beta (IFN beta; 1000 units), as well as interferon alpha (IFN alpha; 1000 units) resulted in roughly a four fold increase in the co-expression of the 42-46 kDa/GD3 antigens as well as the 42-46 kDa/57 kDa antigens. After a 4 day incubation the co-expression of these antigens progressed and IFN alpha treatment had the most pronounced effect, which was 8 fold higher than background for the 42-46 kDa/57 kDa antigens, whereas IFN beta resulted in a five fold antigen upregulation. The pronounced effect of vinblastine on the co-expression of the 42-46 kDa/GD3 antigens (4 fold on day 1 and, 10 fold on day 4) and 42-46 kDa/57 kDa antigens on PANC-1 (5 fold on day 1 and 7 fold over background on day 4) cells can be seen at concentrations as low as 10(-7)M. Colchicine and vincristine dramatically enhanced co-expression of these tumor antigens on day 4 but not on day 1 PANC-1 cells. The expression of these antigens was also found to be cell cycle dependent.

Immunoglobulin repertoire of B lymphocytes infiltrating breast medullary carcinoma.

Tumor specific peptides recognized by T lymphocytes infiltrating solid tumors, as well as the corresponding T cell receptor (TcR) repertoire usage, have been extensively investigated. By contrast, tumor infiltrating B cells and their immunoglobulin (Ig) repertoire have been studied only in a limited number of tumors. The objective of the present study was to determine, whether DNA sequence analysis of the expressed immunoglobulin variable regions of B cells that infiltrate breast cancer, could be used to reveal a potential specific tumor binding capacity of the antibodies. To answer this question, about 200 expressed Ig heavy (VH) and light chain variable gene (VL) regions were cloned, sequenced and comparatively analysed from a typical medullary beast carcinoma (MBC), where the massive B and plasma cell infiltration correlates with favourable prognosis despite of its high grade. The tumor infiltrating B cell Ig heavy and light chain sequences could be classified into clusters, families and subgroups, based on the identity level to germline, showing a pattern of oligoclonality. Some overrepresented clusters could be determined. In the course of a detailed analysis and search in Blastn database, a number of VH and VL sequences showed more than 99% homology to DNA sequences of Ig VH region, with proved tumor antigen binding capacity. Our data suggest, that potential tumor binder Ig VH and VL sequences might be selected using a detailed immunoglobulin variable region analysis. This new approach might have a benefit for further antibody engineering, as difficulties in search for tumor binders by phage library selection might be reduced and the time for selection shortened.

Clinical characteristics and management responses in 85 HIV-infected patients with oral candidiasis.

Eighty-five consecutively seen HIV-positive persons with oral candidiasis were evaluated for clinical characteristics, staging of HIV disease, quantitation of candidal colony formation, and response to systemic antifungal treatment with Nizoral (ketoconazole). Fifty-five had CD4 counts less than 200. There was an inconsistent association between clinical signs, patient symptoms, CD4 counts, and candidal colony-forming units. However, there was a trend toward higher colony-forming unit counts (> 500) in patients with lower CD4 cells (< 200). Sixty-five patients had a complete clinical response to the ketoconazole treatment (200 mg daily for 7 days), even though 81% of posttreatment cultures remained positive. Nonsmokers were more likely to respond to antifungal treatment when compared with smokers, and there was a slight tendency for complete responses when colony-forming unit counts were low. The most common lesion presentation was a combination of the white (pseudomembranous) and red (erythematous) forms. Forty-nine percent had complaints of pain. The variable responses indicated the importance of flexible dose-time and drug considerations in antifungal management. Candida albicans was the predominant species.

Characterization of human IgG1 monoclonal antibody against gangliosides expressed on tumor cells.

A human IgG1.k monoclonal antibody (MAb) designated GMA1 was developed by fusing pooled lymph node lymphocytes from cancer patients with the human lymphoblastoid cell line, SHFP-1. The GMA1 MAb reacted with several melanoma and neuroblastoma cell lines. Normal tissue derived from human brain and tumor-cell lines derived from colon, ovary, and breast were not reactive. FACS analysis performed using live cells demonstrated that the antibody recognizes a cell-surface antigen. Enzyme immunoassay (EIA) and thin layer chromatography (TLC) immunostaining with purified gangliosides indicated that the antibody has specificity for the major tumor associated gangliosides GD3, GM3, and GD2. GMA1 heavy and light chain genes were isolated by RT-PCR and a recombinant derivative of this human antibody was expressed in Chinese hamster ovary (CHO) cells. High-level antibody synthesis and secretion was achieved using a vector designed to maximize expression. FACS analysis and TLC immunostaining indicated recombinant GMA1 reacted with human tumor cell lines and gangliosides GD3, GM3, GD2 in a manner similar to the antibody produced by the hybridoma cell line, demonstrating that the specificity of the antibody was not altered during molecular cloning.

Isotype-directed enrichment of B cells by magnetic beads in the generation of immunoreactive human monoclonal antibodies.

Pooled lymphocytes collected from cancer patients were mixed with a biotinylated murine MAb specific to human IgG4. To this were added streptavidin-conjugated magnetic beads. After magnetically separating the bead-lymphocyte complex, the B cells were washed and fused with the WIL-2 derived human fusion partner, SHFP-1. Subsequently derived human-human hybridomas were screened for IgG4 immunoreactivity to target tumor cell lines. Several hybridomas reacted with a variety of malignant cell types, including melanoma, neuroblastoma, and pancreatic tumor cells. One hybridoma in particular, designated SC-GM4, recognized an antigen by Western blot with an apparent molecular weight of 57 kDa. This facile approach of magnetically separating selected populations of lymphocytes should be relatively simple to apply to other antigens and antibodies to preselect the type, class, and properties of the desired MAb.

Cloning and expression of the human tumor-specific antibody GM4.

The human monoclonal antibody GM4 was generated by fusing pooled lymphocytes from cancer patients with the lymphoblastoid cell line SHFP-1. Immunohistochemical staining of tumor and normal tissue indicated that this human IgG4 antibody preferentially reacted with melanomas and neuroblastomas. In this study, we demonstrate that GM4 recognizes a "vimentin-like" peptide sequence that we have termed AgGM4. To generate a recombinant derivative of this human antibody, we isolated and expressed the complete heavy and light chain genes. The entire coding sequence for both the heavy and light chains was isolated by RT-PCR using a set of degenerate 5' signal sequence specific primers and a 3' constant region primer. High level antibody synthesis and secretion was achieved in Chinese hamster ovary (CHO) cells using a vector designed to maximize expression. Western blot and FACS analysis indicated recombinant GM4 reacted with human tumor cell lines and AgGM4 in a manner similar to the antibody produced by the hybridoma cell line, demonstrating that the specificity of the antibody was not altered during molecular cloning.

A retrospective study of two-stage treatment outcomes assessed with two modified PAR indices.

This retrospective study was undertaken to evaluate the long-term outcome of two-stage (functional/fixed) Class II treatment. A modified peer assessment rating (PAR) was applied to the records of 27 patients who had been recalled an average of 9 years after the completion of the second phase of treatment. UK and US weightings were applied. Analysis of variance identified significant differences among treatment stages. The greatest change in PAR score occurred during the first (functional) phase of treatment. By the end of the second phase, there had been an 83% reduction in PAR score. At recall, however, the PAR scores had increased significantly, due largely to relapse in overjet and in the lower labial segment. These results call into question the ultimate utility of early, two-stage treatment regimens. Although the differences between the UK and USA weightings were smaller than anticipated, the nature of the relapse seen here argues against the American exclusion of the lower labial segment.

Simulation training for dental foundation in oral and maxillofacial surgery - a new benchmark.

Simulation training involves reproducing the management of real patients in a risk-free environment. This study aims to assess the use of simulation training in the management of acutely ill patients for those in second year oral and maxillofacial surgery dental foundation training (DF2s). DF2s attended four full day courses on the recognition and treatment of acutely ill patients. These incorporated an acute life-threatening events: recognition and treatment (ALERT(™)) course, simulations of medical emergencies and case-based discussions on management of surgical inpatients. Pre- and post-course questionnaires were completed by all candidates. A maximum of 11 DF2s attended the course. The questionnaires comprised 1-10 rating scales and Likert scores. All trainees strongly agreed that they would recommend this course to colleagues and all agreed or strongly agreed that it met their learning requirements. All DF2s perceived an improvement in personal limitations, recognition of critical illness, communication, assessing acutely ill patients and initiating treatment. All participants felt their basic resuscitation skills had improved and that they had learned new skills to improve delivery of safety-critical messages. These techniques could be implemented nationwide to address the more complex educational needs for DF2s in secondary care. A new benchmark for simulation training for DF2 has been established.