ANTIBODIES TO SACCHAROMYCES CEREVISIAE and ANTINEUTROPHIL CYTOPLASMIC AUTOANTIBODIES IN INFLAMMATORY BOWEL DISEASE: ASSESSMENT and RELEVANCE IN AN AUSTRALIAN POPULATION USING TWO DIFFERENT ASCA ASSAY KITS

A number of North American and European studies have elucidated a relationship between antibodies to the yeast Saccharomyces cerevisiae (ASCA) and Crohn's disease (CD).
Aims  (1) To ascertain whether this relationship is relevant to Australian patients; (2) To compare the results with two different commercial ASCA kits; (3) To examine the usefulness of this test in combination with perinuclear antineutrophil cytoplasmic autoantibodies (pANCA) for distinguishing Crohn's disease from ulcerative colitis (UC).
Methods  Serum samples were obtained from 28 patients with CD, 27 patients with UC and 22 non-IBD patients presenting for investigation of other gastroenterological illnesses. ASCA IgG and IgA were determined by enzyme immunoassay using the two test kits. pANCA was detected by indirect immunofluorescence.
Results  Using the Medizym test kit, the presence of either IgG or IgA ASCA was 50% sensitive and 93% specific for CD. The QUANTA Lite kit yielded a higher sensitivity of 79% but specificity of 74%. The sensitivity of pANCA for UC was 48% but was 100% specific. Used in combination, ASCA+ve/pANCA–ve was only 50% sensitive but 100% specific for CD using the Medizym kit compared with 79% sensitivity and 93% specificity using QUANTA Lite. The combination of ASCA–ve/pANCA+ve was 41% sensitive and 100% specific for ulcerative colitis using the Medizym kit compared with 30% sensitive and 100% specific using QUANTA Lite.
Conclusions  At least 50% of Australian patients with CD have ASCA detectable in serum, confirming the results of overseas studies. Sensitivity was greater with the QUANTA Lite kit whereas the Medizym kit was slightly more specific. ASCA may aid in the diagnosis of CD. When used in combination with pANCA it may also help distinguish CD from UC in difficult cases.     

CD34+ human marrow cells that express low levels of Kit protein are enriched for long-term marrow-engrafting cells

Using in utero transplantation into fetal sheep, we examined the capability of human bone marrow CD34+ cells fractionated based on Kit protein expression to provide long-term in vivo engraftment. Twelve hundred to 5,000 CD34+ Kit-, CD34+ Kit(low), and CD34+ Kit(high) cells were injected into a total of 14 preimmune fetal sheep recipients using the amniotic bubble technique. Six fetuses were killed in utero 1.5 months after bone marrow cell transplantation. Two fetuses receiving CD34+ Kit(low) cells showed signs of engraftment according to analysis of CD45+ cells in their bone marrow cells and karyotype studies of the colonies grown in methylcellulose culture. In contrast, two fetuses receiving CD34+ Kit(high) cells and two fetuses receiving CD34+ Kit- cells failed to show evidence of significant engraftment. Two fetuses were absorbed. A total of six fetuses receiving different cell populations were allowed to proceed to term, and the newborn sheep were serially examined for the presence of chimerism. Again, only the two sheep receiving CD34+ Kit(low) cells exhibited signs of engraftment upon serial examination. Earlier in studies of murine hematopoiesis, we have shown stage-specific changes in Kit expression by the progenitors. The studies of human cells reported here are in agreement with observations in mice, and indicate that human hematopoietic stem cells are enriched in the Kit(low) population.     

Computer graphics for digitally formatted images

The increasing use of digitally formatted imaging systems requires high-quality interactive gray-scale computer raster graphics systems for the management, display, and analog film recording of digital image and alphanumeric information. These systems are a combination of computer hardware and software and implement a set of graphics protocols. This paper describes a set of interactive graphics protocols that has been developed for clinical use.     

Nutrition support service: role of the clinical dietitian

A study was undertaken to determine: the qualifications necessary to function on a nutrition support team as perceived by clinical dietitians working in this capacity, the actual role of the clinical dietitian on a nutrition support team, the ideal role of the clinical dietitian on a nutrition support team, and the extent to which clinical dietitians perceive differences between the ideal role expectation and actual performance. A questionnaire was developed and sent to a random sample of 300 clinical dietitians listed as members of a nutrition support service. The respondents indicated that the clinical dietitian should have at least a B.S., R.D., and 2 years' prior work experience before assuming responsibility on a nutrition support team. The dietitians indicated that they consistently take and evaluate diet histories and assess energy and protein needs. Moreover, they viewed these tasks as appropriate. They rarely administer or interpret antigen skin tests and do not perceive this as a function of the dietitian. For all other tasks, dietitians indicated that they should perform the duty or responsibility more often. Over half reported that they did not have adequate educational preparation to assume all of the responsibilities of the clinical dietitian on a nutrition support team.     

Virtual colonoscopy with magnetic resonance imaging: In vitro evaluation of a new concept

BACKGROUND & AIMS: Screening for colonic polyps is desirable. A new concept based on cross-sectional and endoscopic analysis of a magnetic resonance (MR) data set is presented. METHODS: Ex vivo autopsy colonic specimens, containing artificially placed polyps, were obtained and filled with a gadolinium-containing solution. Forty-four thin-section MR images were obtained in a 1.5-T MR scanner in 28 seconds. A three- dimensional endoscopic fly-through of these images was rendered. Fly- throughs and two-dimensional cross-sectional images were analyzed by two observers for the presence of polyps. RESULTS: The average sensitivity and specificity for the detection of polyps based on three- dimensional endoscopic MR colon imaging were 87% and 96%, respectively. Analysis of cross-sectional images showed an overall sensitivity and specificity of merely 57% and 84%, respectively. The difference in the interpretation of three-dimensional MR colonoscopy and two-dimensional cross-sections was statistically significant (P < 0.001). With three- dimensional MR colonoscopy, overall sensitivity for detection of polyps measuring < or =5 mm in length and diameter was 70%; for larger polyps, it increased to 95% (P < 0.01). CONCLUSIONS: The feasibility of an MR- based endoluminal assessment of the colon is shown. Minimal invasiveness, lack of radiation exposure, and high in vitro diagnostic accuracy warrant further investigation of this novel concept. (Gastroenterology 1997 Jun;112(6):1863-70)     

Level and direction of hope in cancer patients: an exploratory longitudinal study

Goals

Hope is an important factor to consider when caring for cancer patients as a key component of coping with adversity. The aim of our study was to address the following questions: Is there a difference in level of hope between those patients being curatively and those palliatively treated, and how does this change over time? What are patients’ most important hopes? Better understanding of patients’ hopes may promote more effective patient-centered care.

Materials and methods

Outpatients referred to medical oncology or pain and symptom clinics at the London Regional Cancer Program were surveyed before consultation. The Herth Hope Index (HHI) questionnaire was administered to assess level of hope, and the patients were asked to indicate their highest priority hopes. Qualitative thematic analysis was performed on the responses, and comparison was made between patients treated with curative vs those treated with palliative intent. This survey was repeated 4 months after initial assessment.

Results

Fifty patients were surveyed (29 curative-intent; 21 palliative). Highest priority initial hopes were categorized as follows: cure, other positive health outcomes, emotional well-being, life achievements/return to normalcy, interpersonal goals, other. There was no association noted between treatment intent and choice of highest-priority hope. Follow-up assessments after 4 months revealed no significant differences in the distribution pattern of hopes. There was a statistically significant increase in the HHI over time in curatively treated patients, but none in the combined analysis.

Discussion and conclusion

Our study indicates that patients receiving palliative therapy have a HHI score not significantly different from patients being treated for cure. The hope deemed most important is also similar between groups. Over time, overall hope was maintained or increased even in the presence of a trend towards fewer patients hoping for a cure. These results remind oncologists to explore the experience of hope with all patients to ensure that the subjective needs and goals of the patients are met by the proposed therapies.

     

富血小板血浆对人骨髓间充质干细胞成骨分化的抑制效应

目的:一些理论质疑富血小板血浆对骨前体细胞成骨分化的作用,本实验拟验证富血小板血浆对体外培养的人骨髓间充质干细胞成骨分化的抑制效应。方法:实验于2005-05/11在南方医科大学组织工程试验室(省级)完成。①实验方法:抽取6名健康志愿者髂前上棘骨髓5mL进行体外细胞培养扩增,静脉血10mL以二次离心法制得富血小板血浆。诱导骨髓间充质干细胞时富血小板血浆与骨髓间充质干细胞均来自同一个体。②碱性磷酸酶染色:取第4代骨髓间充质干细胞,分为两组:富血小板血浆组加入富血小板血浆使终浓度为100g/L,单纯血清培养组仅加入等量胎牛血清。培养后第7天进行碱性磷酸酶染色,阳性细胞为胞质中呈现黑色颗粒或块状沉淀。③矿化结节染色:取第4代骨髓间充质干细胞,分组同上。培养后第19天以0.1%茜素红-TrisHcl(pH8.3)37℃下放置30min,矿盐沉积染色阳性为红色。④Cbfa1基因表达:取第4代骨髓间充质干细胞,分组同上。培养后第3,7,12,16天RT-PCR法检测骨髓间充质干细胞Cbfa1基因的表达。⑤形态学观察:实验过程中使用相差显微镜观察各组细胞生长情况及形态学变化。结果:①骨髓间充质干细胞碱性磷酸酶染色结果:培养后第7天,富血小板血浆组碱性磷酸酶阳性细胞数量较单纯血清培养组明显减少,且阳性细胞内灰黑色颗粒也明显减少,为弱阳性。②骨髓间充质干细胞矿化结节染色结果:培养后第19天,单纯血清培养组可见细胞表面有较多的矿盐沉积,但未形成明显的矿化结节。富血小板血浆组细胞表面只有稀少的矿盐沉积。③骨髓间充质干细胞cbfa1mRNA的表达:培养后第3,7,12,16天,随着培养时间的延长单纯血清培养组与富血小板血浆组cbfa1基因表达量均逐渐增高,同一时间点两组间cbfa1基因的表达基本相似。④骨髓间充质干细胞形态学变化:富血小板血浆组骨髓间充质干细胞增殖旺盛,细胞达到单层汇合的时间较单纯血清培养组明显缩短。单纯血清培养组细胞在完全汇合后开始出现聚合现象(14～16d),但趋向性不明显,未完全形成团簇;富血小板血浆组细胞在完全汇合后未出现聚合现象,细胞密集生长。培养初期两组细胞以梭形为主,多角形细胞较少,培养至14～16d单纯血清培养组多角形细胞较富血小板血浆组增多。结论:富血小板血浆可抑制人骨髓间充质干细胞碱性磷酸酶的分泌与矿盐沉积,对人骨髓间充质干细胞成骨分化的直接效应是抑制其分化。     

Interleukin-3 treatment of rhesus monkeys leads to increased production of histamine-releasing cells that express interleukin-3 receptors at high levels

To understand the hematopoietic and nonhematopoietic responses to interleukin-3 (IL-3), expression of cell-surface IL-3 receptors (IL-3R) was examined on bone marrow (BM) cells and peripheral blood (PB) cells of rhesus monkeys during the course of in vivo IL-3 treatment. Whereas IL-3R expression is low in untreated monkeys, IL-3 administration led to a gradual increase in both low- and high-affinity binding sites for IL-3. This increase reflected the total number of cells expressing IL- 3Rs, as detected by flow cytometry using biotinylated IL-3. Most of these IL-3R+ cells in both BM and PB could be characterized as basophilic granulocytes that contained high levels of histamine. In contrast to the effect on these differentiated cells, IL-3 administration did not significantly alter the low level IL-3R expression on immature, CD34+ cells. Further flow cytometric analysis using biotinylated growth factors showed that the IL-3R+ basophils also expressed receptors for granulocyte-macrophage colony-stimulating factor (GM-CSF), but not for IL-6 or Kit ligand. These findings indicated that the IL-3R+ cells included neither monocytes, which express GM-CSFRs and IL-6Rs abundantly, nor mast cells, which express c- kit. By combining flow cytometric and Scatchard data, it was calculated that the basophils contain as many as 1 to 2 x 10(3) high-affinity IL- 3Rs and 15 to 30 x 10(3) low-affinity sites. The finding that in vivo IL-3 treatment leads to the production of large numbers of cells that express high levels of IL-3R and are capable of producing histamine provides an explanation for the often severe allergic reactions that occur during prolonged IL-3 administration. It also indicates that IL- 3, in addition to its direct effects on hematopoietic cells, may also stimulate hematopoiesis through the release of secondary mediators such as histamine by IL-3-responsive mature cells.