bluuebunny/crossref_metadata_2025 huggingface dataset:
Patterns of Dwarf expression and brassinosteroid accumulation in tomato reveal the importance of brassinosteroid synthesis during fruit development
Authors: Montoya Teresa , Nomura Takahito , Yokota Takao , Farrar Kerrie , Harrison Kate , Jones Jonathan G. D. , Kaneta Tsuyoshi , Kamiya Yuji , Szekeres Miklos , Bishop Gerard J.
Published: March 2005

Brassinosteroids (BRs) are essential for many physiological functions in plants, however little is known concerning where and when they are synthesized. This is especially true during flower and fruit production. To address this we have used a promoter‐GUS reporter fusion and RT‐PCR to determine the relative expression levels of the tomato Dwarf (D) gene that encodes a BR C‐6 oxidase. In young seedlings GUS reporter activity was observed mainly in apical and root tissues undergoing expansion. In flowers GUS activity was observed in the pedicel joints and ovaries, whereas in fruits it was strongest during early seed development and was associated with the locular jelly and seeds. RT‐PCR analysis showed that tissue‐specific expression of Dwarf mRNA was consistent with that of the Dwarf:GUS fusion. In good correlation with the high local Dwarf activity, quantitative measurements of endogenous BRs indicated intense biosynthesis in developing tomato fruits, which were also found to contain high amounts of brassinolide. Grafting experiments showed the lack of BR transport indicating that BR action occurs at the site of synthesis.
Seroprevalence of dengue antibodies, annual incidence and risk factors among children in southern Vietnam
Authors: Thai Khoa T. D. , Binh Tran Quang , Giao Phan Trong , Phuong Hoang Lan , Hung Le Quoc , Nam Nguyen Van , Nga Tran Thanh , Groen Jan , Nagelkerke Nico , de Vries Peter J.
Published: March 2005

Dengue is highly endemic in southern Vietnam and all four serotypes of dengue virus have already been identified. To determine the age‐specific prevalence of dengue and associated risk factors, we conducted a serological study at two primary schools and assessed risk factors by analysing children's questionnaires and household surveys. Sera were collected from 961 primary schoolchildren in Binh Thuan Province and tested for the presence of dengue virus serum antibodies using an indirect immunoglobulin G (IgG) enzyme‐linked immunosorbent assay (ELISA). The antibody prevalence of the total population was 65.7% (n = 631) which increased from 53.0 to 88.2% with age. The annual incidence of a first dengue infection, estimated by binary regression of the seroprevalence by age, was 11.7%. Interestingly, the prevalence of dengue IgG antibodies was significantly higher in children who confirmed using a pit latrine (RR 1.467, 95% CI: 1.245–1.730) and whose domestic environment contained discarded cans (RR 1.238, 95% CI: 1.042–1.470) and pigs (RR 1.228, 95% CI: 1.002–1.504). The epidemiology of dengue in southern Vietnam is stable with a constantly high annual incidence of first infections. Transmission occurs mainly peri‐domestically, which has important public health implications.
Changes in the geographical distribution of tuberculosis patients in Veracruz, Mexico, after reinforcement of a tuberculosis control programme
Authors: Jacobson Lara M. , de Lourdes Garcia‐Garcia Ma. , Hernandez‐Avila Juan Eugenio , Cano‐Arellano Bulmaro , Small Peter M. , Sifuentes‐Osornio Jose , Ponce‐de‐Leon Alfredo
Published: March 2005

Objective  Geographical information systems are valuable tools for studying tuberculosis (TB) epidemiology, but have been underused for the evaluation of TB control programs. We aimed to compare the geographical distribution of TB before and after the five elements of directly observed therapy, short course (DOTS) were strengthened in a Health Jurisdiction in Mexico in response to recommendations made by the WHO Global Tuberculosis Program.

Methods  All consenting persons detected by passive case finding between March 1995 and February 2000 who were confirmed to have acid‐fast bacilli (AFB) in sputum underwent clinical and laboratory evaluation. A TB ‘episode’ was defined as the period of time between bacteriological diagnosis and treatment discharge by the local control programme. Distances of TB episodes from the nearest urban centre were determined according to recent transmission and multidrug resistance (MDR).

Results  During the first half of the study period, MDR episodes were located a median distance of 1.24 km from urban centres, which did not differ significantly from non‐MDR episodes (1.14 km, P = 0.56). In contrast, the median distance of MDR episodes increased 55% to 1.92 km during the second half, which placed them significantly further away out than non‐MDR episodes (1.08 km, P = 0.01). No changes in location were detected for recent transmission.

Conclusion  While reinforcing the TB control programme reduced the incidence of MDR, the remaining episodes were located in poorer and more remote areas.
Familial cutaneous collagenoma: report of an affected family
Authors: Amato L. , Mei S. , Gallerani I. , Moretti S. , Cipollini E. M. , Palleschi G. M. , Fabbri P.
Published: March 2005

A 44‐year‐old white woman was admitted to our hospital for the investigation of many roundish, pale‐colored, nodular lesions of different sizes, ranging from a few millimeters to some centimeters, symmetrically grouped on the shoulders (Fig. 1), trunk, upper limbs, and abdomen. The lesions first appeared at 20 years of age on the upper chest around the clavicular region, and thereafter gradually increased on the trunk and upper limbs; they were completely asymptomatic, appearing on normal skin without a previous history of inflammation and/or trauma. No scarring or atrophy was evident. The smaller lesions exhibited a miliaria‐like shape, being evident in particular on stretching of the skin; the larger ones showed a firm‐elastic density, no adhesion to surrounding tissue, and deep localization in the skin. No hypopigmented macules or sebaceous adenoma‐like lesions were present. Skin biopsy of a nodular lesion from the right shoulder revealed a normal epidermis and a dermis filled with bundles of dense and coarse collagen fibers (Fig. 2). Elastic fibers appeared thin and decreased in number under Verhoeff–van Gieson stain (Fig. 3). A diagnosis of cutaneous collagenoma was proposed. Routine laboratory investigations were within the normal range. Chest and skeleton (hands, pelvis, and long bones) X‐ray, electrocardiogram, and echocardiogram showed no abnormality.

Nodular papules localized on the right shoulder

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Histology of a lesion taken from the right shoulder. The picture demonstrates dense and coarse collagen bundles (hematoxylin and eosin stain; original magnification, × 200)

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Elastic fibers are clearly diminished in number, thin, and fragmented (Verhoeff–van Gieson stain; original magnification, × 400)

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In order to evaluate a possible familial cutaneous collagenoma, the 19‐year‐old son of the patient was also examined. He had a 3‐month history of numerous, asymptomatic, skin‐colored nodules, ranging from 0.5 to 2 cm, on the upper back. Skin biopsy of a nodular lesion showed the same features as observed in the mother's biopsy. Routine blood and serum tests and X‐ray examinations were within normal limits. Electrocardiogram revealed an early R‐wave transition, and echocardiogram showed a left ventricular dilation with thickened aorta; aortic steno‐insufficiency was diagnosed under cardiologic examination. The diagnosis of familial collagenoma was confirmed, and similar cutaneous lesions were found in the mother's grandmother and aunt.
Localization of haptoglobin in normal human skin and some skin diseases
Authors: Li Ping , Gao Xing‐Hua , Chen Hong‐Duo , Zhang Yaqin , Wang Yan , Wang He , Wang Yakun , Xie Yong
Published: March 2005

Background  Haptoglobin (Hp) is an acute‐phase reactant, known to be produced mainly in the liver. Haptoglobin can also be detected in the cytoplasm of normal epidermal Langerhans cells (LCs), and can prevent their functional maturation. The synthesis of Hp in skin cells has not been well studied.

Methods  We examined Hp expression at mRNA and protein levels by in situ hybridization and immunohistochemistry, respectively, in normal human skin and in the skin of patients with psoriasis, lichen planus, erythroderma, seborrheic keratosis, verruca vulgaris, basal cell carcinoma, systemic lupus erythematosus, pemphigus and bullous pemphigoid.

Results  (1) Haptoglobin mRNA was expressed in the epidermal keratinocytes (KCs), the epithelial cells of hair follicles, sebaceous glands and eccrine glands in normal skin and all dermatoses investigated. (2) Whereas compared with normal skin, the Hp mRNA in KCs of patients with psoriasis, lichen planus, erythroderma, seborrhoea keratosis and verruca vulgaris was significantly intensified, it was weaker in patients with systemic lupus erythematosus, pemphigus and bullous pemphigoid. (3) Haptoglobin protein only stained positively in some KCs of patients with psoriasis, lichen planus and erythroderma. (4) Although some but not all epidermal LCs were positively stained with anti‐Hp antibody in normal skin and in skin samples from all patients, the ratios of Hp‐positive LCs/total LCs were significantly higher in those diseases with intensified Hp mRNA in KCs.

Conclusions  Skin is another extrahepatic organ where Hp can be synthesized by KCs. The expression of Hp mRNA in KCs and the Hp protein in both LCs and KCs appears to be correlated with the amount of inflammation, which might indicate that skin itself is involved in down‐regulating the local inflammatory reaction by KC‐synthesized Hp.
Pilot trial: Pioglitazone versus placebo in patients with plaque psoriasis (the P6)
Authors: Shafiq Nusrat , Malhotra Samir , Pandhi Promila , Gupta Monica , Kumar Bhushan , Sandhu Kanial
Published: March 2005

Background  Disordered differentiation and hyperproliferation of keratinocytes with inflammation are the hallmarks of psoriasis. Ligand activation of peroxisome proliferator receptor‐γ (a class of nuclear receptors) by thiazolidinediones can normalize the histologic features of psoriasis.

Method  In a 10‐week, double‐blind, randomized, placebo‐controlled, parallel‐group study, 70 patients with moderate to severe psoriasis received one of the following treatments: pioglitazone 15 mg, pioglitazone 30 mg or placebo. Efficacy was evaluated by observing the change in the psoriasis area and severity index (PASI) after 10 weeks of treatment.

Results  There was a dose‐dependent improvement in psoriasis. Median PASI scores at the end of 10 weeks were significantly reduced in the pioglitazone treatment groups as compared to the placebo‐treated group. The psoriasis lesions cleared in more than 40% of patients treated with pioglitazone as compared to 12.5% of those with placebo. The percentage reduction in mean PASI scores was 21.6%, 41.1% and 47.5% in the placebo, pioglitazone 15 mg, and 30 mg groups, respectively. No serious adverse events were detected.

Conclusion  This is the first report from a controlled trial demonstrating that pioglitazone could be considered as an efficacious and safe agent for the treatment of plaque psoriasis. The optimum dose and duration of pioglitazone therapy remain to be determined.
Survival of adult islet grafts from transgenic pigs with N‐acetylglucosaminyltransferase‐III (GnT‐III) in cynomolgus monkeys
Authors: Komoda Hiroshi , Miyagawa Shuji , Omori Takeshi , Takahagi Yoichi , Murakami Hiroshi , Shigehisa Tamotsu , Ito Toshinori , Matsuda Hikaru , Shirakura Ryota
Published: March 2005

Abstract:  Background:  Because of a severe shortage of human donor pancreases, pig islets are considered to be an attractive donor source. Our previous in vitro study revealed that adult pig islets have strong non‐Galα1‐3Galβ1‐4GlcNAc‐R (α‐Gal) antigenicity, including the Hanganutziu‐Deicher (H‐D) antigen, especially in N‐linked sugars. In this study, the issue of whether islets from N‐acetylglucosaminyltransferase‐III (GnT‐III) transgenic pigs can prolong their survival in cynomolgus monkeys was examined.

Methods:  Adult pig islets were isolated from transgenic pigs with GnT‐III and wild‐type genes. GnT‐III enzyme activity in pig islets was measured by high performance liquid chromatography (HPLC). The antigenicity of the islets to human natural antibodies was examined by flow cytometry. Pig islets were transplanted under the kidney capsule of streptozotocin‐induced diabetic monkeys. After transplantation, blood samples were obtained and plasma insulin levels were monitored on a daily basis.

Results:  While GnT‐III was barely expressed in wild‐type islets, it was expressed at high levels in islets from transgenic pigs, and xenoantigenicity was significantly reduced. There was a trend for islets isolated from GnT‐III‐transgenic pigs to survive longer than those from wild‐type pigs in cynomolgus monkeys (wild type: 1, 1, and 3 days; GnT‐III: 1, >3, 4 and 5 days). Humoral and histological studies indicated up‐regulated anti‐pig islet antibodies and a relatively high deposition in islet grafts from wild‐type pigs, respectively.

Conclusion:  A reduction in xenoantigenicity by GnT‐III may have prolonged the survival of porcine islets, suggesting the importance of non‐α‐Gal and non‐H‐D antigens, as they relate to N‐linked sugars in the early rejection of porcine islets in the monkey. This approach may be useful in the clinical xenotransplantation of islets in the future.
Improved method for isolation of porcine neonatal pancreatic cell clusters
Authors: Valdés‐González Rafael A. , Silva‐Torres María L. , Ramírez‐González Briceyda , Terán Luis , Ormsby Christopher E. , Ayala‐Sumuano Jorge T.
Published: March 2005

Abstract:  Based on the described methods for the isolation of neonatal pancreatic cell clusters (NPCCs), we have developed modifications in order to improve their quality, functionality, and process reproducibility in the isolation technique, for potential use in research. In addition, we indicate techniques for describing yield, functionality, viability and purity of the NPCCs.

Methods:  Purity of the NPCCs was determined through dithizone staining and subjected to image analysis. Viability and apoptosis was measured through flow cytometry with propidium iodide and annexin, respectively. NPCC functionality was measured through a static glucose stimulation test.

Results:  We developed a high‐yield reproducible technique that had 81 279.55 ± 18 257.05 IEQ/g of pancreas at 4 days of culture, with a 94% viability and an 88 ± 2.73% purity. Stimulation index from the glucose stimulation test was >10.

Conclusion:  The technique allowed us to obtain NPCC with optimal viability, functionality, purity, and endurance for use in research.
The activity of inducible nitric oxide synthase in rejected skin xenografts is selectively inhibited by a factor produced by grafted cells
Authors: Holáň Vladimír , Pindjáková Jana , Zajícová Alena , Krulová Magdaléna , Železná Blanka , Matoušek Petr , Svoboda Petr
Published: March 2005

Abstract:  Background:  Production of nitric oxide (NO) by graft infiltrating macrophages has been suggested as an important effector mechanism of allograft rejection. Expression of the gene for the inducible NO synthase (iNOS) and the production of NO in rejected graft has been demonstrated in various models of allotransplantation. However, whether NO plays a role in rejection of skin xenografts has not been documented.

Methods:  Explants of rejected skin allografts or xenografts (rat to mouse) were cultivated in vitro and the production of NO, interleukin (IL)‐2, IL‐4, IL‐10 and interferon‐γ (IFN‐γ) by graft infiltrating cells was determined by the Griess reaction or ELISA. Effects of supernatants from cultures of xenograft explants on the expression of gene for iNOS, accumulation of iNOS protein and NO production were determined by RT‐PCR or Western blots. Molecular mass of the factor with the suppressive activity was characterized by filtration on chromatography Sephacryl S‐200 Superfine column. In addition, the effects of 2‐amino‐5,6‐dihydro‐6‐methyl‐4H‐1,3‐thiazine (AMT), a selective iNOS inhibitor, on survival of skin xenografts were tested.

Results:  While explants of rejected mouse skin allografts produced substantial amounts of NO, undetectable or only very low levels of NO were found in supernatants from cultured rat skin xenografts. Cocultivation of bacterial lipopolysaccharide (LPS)‐stimulated mouse macrophages which produce high quantities of NO, with pieces of rejected xenografts, but not of syngeneic grafts, allografts or normal rat skin, completely inhibited production of NO. Production of IL‐6 and IL‐10 by LPS‐stimulated macrophages was not inhibited under the same conditions. The inhibition of NO production was mediated by a factor which was produced by rejected rat xenograft and which was eluted from chromatography Sephacryl S‐200 Superfine column in a fraction representing a molecular mass of 67 kDa. The factor did not inhibit the expression of the gene for iNOS, reduce the level of iNOS protein in stimulated macrophages, or function as a scavenger of NO. Rather, the factor inhibited the function of iNOS. The finding that NO does not play an important role during rejection of skin xenografts is supported by the observation that treatment of graft recipients with AMT, a specific iNOS inhibitor, did not enhance xenograft survival, while the same treatment resulted in prolongation of survival of skin allografts.

Conclusion:  The results thus demonstrate that a 67‐kDa molecule produced by rejected rat skin xenografts selectively inhibits iNOS activity in graft infiltrating macrophages. We suggest that NO does not play a significant role in rejection of skin xenografts as it does in the case of allograft rejection.
Use of pigs for xenotransplantation: the speciesism by proxy syndrome
Authors: Ravelingien An
Published: March 2005

Abstract: Background: When questioning whether it is acceptable to use pigs as sources of xenografts, focus easily shifts to demonstrations that it is more acceptable to exploit pigs rather than primates. Both practical and ethical reasons against the use of primates do not stand when applied to the use of pigs. Consequently, use of pigs is held to be justified.

Methods: In this paper, I examine the weaknesses that shelter within three ethical arguments commonly rehearsed in defense of choosing the pig as source animal: (a) that the use of pigs for human purposes is embedded in a long tradition; (b) that pigs are not an endangered species; and (c) that they do not share the cognitive and emotional capacities with humans to the same extent that primates do.

Results: All of the arguments rest on presumptions that are themselves in need of further discussion or clarification.

Conclusion: Other approaches are necessary in justifying use of pigs as sources of xenografts.
Porcine complement regulators protect aortic smooth muscle cells poorly against human complement‐induced lysis and proliferation: consequences for xenotransplantation
Authors: Capey Steven , Van Den Berg Carmen W.
Published: March 2005

Abstract:  Background:  Accelerated atherosclerosis after transplantation has been observed and is characterized by smooth muscle cell proliferation in the graft. Porcine cells are frequently used in models of atherosclerosis and porcine organs are considered for use in transplantation. Complement (C) activation is known to play a major role in rejection of xenografts and is also considered to play a role in the development of atherosclerosis. The aim of this study was to investigate the expression and function of membrane bound regulators of complement (CReg) on porcine aortic smooth muscle cells (PASMC).

Methods:  The PASMC were assessed for expression of CReg and susceptibility to lysis by human C by flow‐cytometry. The effect of various cytokines on CReg expression and C‐susceptibility was investigated. The ability of human C to induce cell proliferation was assessed using the Alamar blue assay.

Results:  The PASMC only express the CReg membrane cofactor protein (MCP) and CD59 on their cell surface. MCP expression was increased by interleukin (IL)‐4. In contrast to porcine aortic endothelial cells (PAEC), PASMC were found to be surprisingly sensitive to C‐mediated lysis, mainly due to a low level of expression of CD59. Human C‐induced proliferation of PASMC, which was dependent on complete membrane attack complex (MAC) formation.

Conclusions:  Endogenously expressed CReg on PASMC poorly protect these cells to human C. Human C can induce proliferation of PASMC. In order to prevent accelerated atherosclerosis in porcine xenografts, increased levels of CReg not only have to be obtained on the endothelial cells but also on the smooth muscle cells.
Co‐stimulation blockade targeting CD154 and CD28/B7 modulates the induced antibody response after a pig‐to‐baboon cardiac xenograft
Authors: Wu Guosheng , Pfeiffer Steffen , Schröder Carsten , Zhang Tianshu , Nguyen Bao N. , Lea William , Kelishadi Sean , Atkinson James B. , Schuurman Henk‐Jan , White David J. G. , Azimzadeh Agnes M. , Pierson Richard N.
Published: March 2005

Abstract: Background: The induced antibodies against Galα1,3Gal (Gal) and non‐Gal epitopes may contribute to delayed xenograft rejection (DXR). We asked whether blockade of the CD40/CD154 and CD28/B7 co‐stimulatory pathways modulates the baboon elicited antibody response to pig Gal and non‐Gal antigens.

Methods: Eighteen baboons received heterotopic heart transplants from pigs transgenic for human decay‐accelerating factor (n = 13) or membrane cofactor protein (n = 5). Ten reference ‘‘conventional therapy’’ animals received cyclosporin A, cyclophosphamide and mycophenolate mofetil, with (n = 4) or without (n = 6) anti‐CD20. Eight ‘‘co‐stimulation blockade’’ animals received anti‐CD154 mAb (IDEC‐131) and anti‐thymocyte globulin, with (n = 4) or without (n = 4) anti‐CD20; two of these animals also received CTLA4‐Fc. Anti‐αGal IgG and IgM, anti‐non‐Gal antibodies and graft histology were assessed serially.

Results: Excluding two early graft failures, median graft survival with conventional therapy was 15 days (range 6 to 36 days, n = 8). Anti‐Gal IgG antibody remained low through day 6 to 10, only one graft failure was accompanied by significant rise in anti‐Gal IgG, and the anti‐non‐Gal response was weak (n = 2) or absent (n = 7). However many recipients succumbed with infection (n = 4) or coagulopathy (n = 2); DXR and ICOS+ T cells were prevalent in long‐surviving grafts. With co‐stimulation blockade, excluding three early graft failures, median graft survival was 7 days (range 6 to 11 days, n = 5). This regimen was very well tolerated, but increased anti‐Gal antibody titer within 14 days was associated with graft failure in four of six animals. Although an anti‐non‐Gal response was present in three of six animals during IDEC‐131 monotherapy (one strong, two weak), it was absent in both cases with additional CTLA4‐Fc treatment.

Conclusions: As used here, CD154 blockade alone does not completely prevent induction of Gal and non‐Gal anti‐pig antibodies. Our preliminary data suggest that other co‐stimulation pathways, including CD28/B7 and ICOS, are sufficient to mediate high‐titer anti‐non‐Gal antibody to porcine antigens in baboons, and contribute significantly to the pathogenesis of DXR.
Antibody‐ and complement‐independent phagocytotic and cytolytic activities of human macrophages toward porcine cells
Authors: Ide Kentaro , Ohdan Hideki , Kobayashi Takaaki , Hara Hidetaka , Ishiyama Kohei , Asahara Toshimasa
Published: March 2005

Abstract:  Background:  It has been speculated that host macrophages contribute to rapid clearance of transplanted xenogeneic cells. To address such a possibility, phagocytotic and cytolytic activities of human macrophages toward xenogeneic porcine cells were evaluated in vitro in the absence of antibodies and complement factors.

Methods:  Human peripheral monocyte‐derived macrophages (P‐macrophages) and reticulo‐endothelial macrophages (RE‐macrophages) were obtained from volunteers’ peripheral blood and from the perfusion effluents of liver allografts for transplantation, respectively. 5‐(and 6‐) carboxyfluorescein diacetate succinimidyl ester (CFSE)‐labeled human autologous red blood cells (auto‐RBCs), ABO‐incompatible RBCs (incom‐RBCs) and xenogeneic porcine RBCs (xeno‐RBCs) were incubated with the human macrophages; subsequently, the macrophages that had phagocytosed the RBCs could be identified as CFSE positive cells by FCM analyses and confocal microscopy. Cytolytic activity was quantified by calculating levels of lactate dehydrogenase in each culture supernatant.

Results:  Human RE‐macrophages spontaneously phagocytosed and had a remarkable cytolytic activity toward xeno‐RBCs, but not toward auto‐RBCs or incom‐RBCs. Elimination of α‐galactosyl xenoantigen (α‐Gal) epitopes on xeno‐RBCs did not prevent phagocytotic or cytolytic activity of RE‐macrophages.

Conclusions:  These findings indicate phagocytotic and cytolitic activities of human macrophages toward porcine cells are initiated by a factor other than α‐Gal in a mechanism independent of antibody/complement opsonization.
Reluctance of French patients with type 1 diabetes to undergo pig pancreatic islet xenotransplantation
Authors: Deschamps Jack‐Yves , Roux Françoise A. , Gouin Edouard , Saï Pierre
Published: March 2005

Abstract: Introduction: Type 1 diabetes could possibly be treated by transplantation of pig pancreatic islets. In addition to medical difficulties and ethical problems, social hurdles may need to be overcome. We have evaluated the attitude of patients with type 1 diabetes to the xenotransplantation of pig pancreatic islets and to the potential risks associated with such treatment.

Methods: A survey of 214 patients with type 1 diabetes was carried out in France based on a multiple‐choice questionnaire.

Results: At first, 52.0% of these patients indicated that they would agree to receive pig islet xenografts. The main sources of reluctance were the ‘‘risk of disease transmission’’ (55.5%) and ‘‘risks not yet identified’’ (48.7%). After they were told of the risk of cancer or infection associated with immunosuppression, 74.9% of the respondents chose to refuse the transplant, compared with 48.0% before they heard of such risks. A 68.1% would refuse the xenotransplant if it would not exempt them completely from being treated by insulin injections. Discontinuing insulin injections was the most important priority for diabetic patients (73.5%), rather than limitation of diabetes‐related complications (52.5%) or increase in life expectancy (44.0%). After they were informed of all of the risks associated with the procedure, 70.5% of the respondents decided they would rather not take any risks, and said they would refuse pig islet transplantation.

Conclusion: When diabetic patients learned about potential infectious risks and other risks associated with immunosuppression, reluctance to undergo xenotransplantation gained in significance or even led to refusal of the procedure.
Candida, still number one – what do we know and where are we going from there?
Authors: Segal Esther
Published: March 2005

Candida species are considered as the most important fungal human pathogens, causing a variety of clinical entities, ranging from superficial, cutaneous‐mucosal to deep‐seated and disseminated infections. A vast body of scientific literature, has been accumulated on these pathogens. A review of the literature and topics for further research are discussed.
Clinical evaluation of Aspergillus‐PCR for detection of invasive aspergillosis in immunosuppressed patients
Authors: Lass‐Flörl Cornelia , Gunsilius E. , Gastl G. , Freund M. , Dierich M. P. , Petzer A.
Published: March 2005

We evaluated the utility of Aspergillus PCR as a tool in diagnosing invasive aspergillosis in patients at risk. Aspergillosis was assessed according to the European Organization for Research and Treatment of Cancer (EORTC), Mycosis Study Group definitions. Nine and seven patients with proven and probable aspergillosis respectively were evaluated. Whole blood samples prior (n = 41) and during antifungal treatment (n = 67), and tissue specimens (n = 9) and/or bronchoalveolar lavage fluids (n = 7) were investigated. In patients with proven infections, the sensitivities of PCR of lung samples were 100%, of blood samples prior treatment were 66%, and during treatment 55%. Clearance of fungal DNA from blood was associated with resolution of clinical symptoms in two of four patients with proven infection. Consecutive positive PCR results for Aspergillus are fatal as two of five patients died. In patients with probable infections, the sensitivities of PCR of lung fluids were 85%, of blood samples prior treatment were 57%, and during treatment 42%. The benefits of PCR diagnosing and screening of whole blood are limited if sampling takes place once treatment has started. The performance of Aspergillus PCR from tissue samples should be recommended in addition to microscopic examination and culture technique for sensitive detection of fungal infection.
Pilze im Mediastinum: Selten aber relevant
Authors: Kujath P. , Scheele J. , Esnaashari H. , Shekarriz H. , Bouchard R.
Published: March 2005

Pilzinfektionen des Mediastinums sind selten. Es lassen sich fünf Formen differenzieren: (i) die hämatogene Pilzinfektion des Mediastinums nach einer Fungämie; (ii) die postoperative Pilzinfektion nach Sternotomie in kardio‐chirurgischer Intention; (iii) als Sonderform der descendierenden nekrotisierenden Mediastinitis (DNM); (iv) die Infektion des Mediastinums per continuitatem; (v) die Pilzinfektion des hinteren Mediastinums nach Ösophagusperforation. In der Klinik für Chirurgie, UK S‐H, Campus Lübeck wurden in 10 Jahren fünf Patienten mit einer Mediastinitis behandelt, die durch Pilze induziert war. Drei Mal handelte es sich um eine descendierende nekrotisierende Mediastinitis, ein Mal um eine Pilzinfektion nach Pseudarthrose des Sternums (postoperativ nach Kardiochirurgie). Der Sektionsbefund einer weiteren Patientin mit einer Lungenaspergillose zeigte eine vollständige Durchsetzung des Mediastinums mit Aspergillus fumigatus.
Intraabdominelle Pilzinfektionen
Authors: Boos C. , Kujath P. , Bruch H. ‐P.
Published: March 2005

The incidence of invasive mycoses in patients undergoing abdominal surgery amounts to approximately 8% and shows an upward trend in epidemiological studies. The lethality of these systemic mycoses, which are mostly based on Candida infections constitutes up to 60%. The development of a sytemic mycosis is marked by exogenic, endogenic and iatrogenic risk factors and typically displays tissue invasion after an initial fungal contamination or systemic dissemination via fungal sepsis. Fungal peritonitis is generally a monoinfection with Candida spp., where Candida albicans outweighs in 70% of cases. Aspergillus spp. are only detected abdominally in rare cases. The histological verification of a fungal invasion is regarded as proof of the existence of an invasive mycosis, but typical macroscopic findings with corresponding cultural findings can also confirm the diagnosis. Systemic mycosis requires an early initiation of a consistent antimycotic therapy as well as definitive surgical eradication of the focus in order to reduce high lethal rate. Resistances or incorrect dosages can be validated objectively by means of histological monitoring of the antimycotic therapy, thus affording early recognition of the need to change the substance class.
Invasive Pilzinfektionen bei Patienten nach Lebertransplantation
Authors: Fischer L. , Sterneck M.
Published: March 2005

Fortschritte bei chirurgischer Technik, Immunsuppression und medizinischem Management haben zu einer wesentlichen Verbesserung der Ergebnisse nach Lebertransplantation (LTx) geführt. Pilzinfektionen stellen bei LTx‐Patienten allerdings nach wie vor schwerwiegende Komplikationen mit signifikant verringertem Überleben dar. Der überwiegende Anteil von Pilzinfektionen nach LTx wird durchCandidaspp. hervorgerufen, was sich aus dem großen abdominalchirurgischen Eingriff erklärt.Aspergillus‐Infektionen stellen die zweithäufigste Pilzinfektion dar, während andere Erreger wiePneumocystis,Cryptococcusund Zygomyzeten nur in Einzelfällen eine Rolle spielen. Die hohe Letalität von invasiven Pilzinfektionen bei LTx‐Patienten ist durch den Schweregrad der Grunderkrankung sowie Schwierigkeiten bei Diagnose und medikamentöser Therapie bedingt. Mit den derzeitigen mikrobiologischen Testverfahren ist eine frühe und zuverlässige Diagnose nicht möglich. In der Therapie stellte Amphotericin B bislang das Mittel der Wahl dar, wobei Nebenwirkungen die Anwendung limitieren. Fluconazol weist eine größere therapeutische Breite und geringe Arzneimittelinteraktionen auf. Nachteile sind fehlende Wirksamkeit aufAspergillus‐Spezies und die Selektion resistenterCandida‐Stämme. Eine Weiterentwicklung stellen aufgrund deutlich geringerer Toxizität und breitem Wirkspektrum neue Azolderivate (Voriconazol) und Echinocandinderivate (Caspofungin) dar. Durch die Identifikation prognostischer Faktoren für die Entwicklung von Pilzinfektionen bei LTx‐Patienten lassen sich Hochrisikogruppen definieren. Eine medikamentöse Prophylaxe bzw. eine präemptive Behandlung bei solchen Risikopatienten kann zu einer Verbesserung der klinischen Ergebnisse führen.
Pilzinfektion bei hämorrhagisch‐nekrotisierender Pankreatitis: Risikofaktoren, Inzidenz, Therapie
Authors: Kujath P. , Rosenfeldt M. , Esnaashari H. , Scheele J. , Bouchard R.
Published: March 2005

Die hämorrhagisch‐nekrotisierende Pankreatitis kann zu einem schweren, intensivpflichtigen Krankheitsbild mit Multiorganversagen führen. Für Patienten mit infizierten Nekrosen wird im allgemeinen die Indikation zur operativen Revision gestellt. Die chirurgische Therapie besteht in einem Ausräumen der Nekrosen (Nekrosektomie). Dadurch wird eine mikrobiologische Aufarbeitung möglich. Die häufigsten nachgewiesenen Erreger sind Enterobacteriaceae. In der Literatur liegt der Anteil der Pilzinfektionen zwischen 15 und 30%. Seit 1996 wurden 73 Patienten mit schwerer nerotisierender Pankreatitis an unserer Klinik behandelt. Bei 50 Patienten (68,5%) ließen sich im Verlauf Pilze nachweisen. Die Letalitätsrate betrug 62%.
Therapie von Mykosen im Rahmen einer kombinierten Pankreas–Nieren‐Transplantation
Authors: Bouchard R. , Hennings L. , Jungbluth Th. , Klempien I. , Kujath P. , Bruch H.‐P.
Published: March 2005

Ein Typ I‐Diabetiker hat nach Entwicklung einer dialysepflichtigen Nephropathie eine Lebenserwartung, die rapide absinkt. Nach drei Jahren Dialyse leben im Durchschnitt nur noch 40% der Typ I‐Diabetiker. Den einzigen Ausweg aus dieser unheilbaren Stoffwechselsituation stellt bislang nur die kombinierte Pankreas–Nierentransplantation dar. Nach einer solchen ist jede Pilzbesiedlung und insbesondere die Pilzinfektion eine ernste Bedrohung für das Leben des Patienten und bedarf einer umgehenden Therapie. Dabei muss zwischen einer oberflächlichen Kolonisation und einer invasiven Mykose unterschieden werden. In Abhängigkeit vom Patientenzustand kann bei ausbleibendem antimykotischem Therapieerfolg die Entfernung des Organs zur Rettung des Patientenlebens notwendig werden.
Surveillance invasiver Fadenpilzmykosen in lungentransplantierten Patienten: Effekt antimykotischer Prophylaxe mit Itraconazol und Voriconazol
Authors: Mattner Frauke , Chaberny Iris F. , Weißbrodt H. , Fischer S. , Gastmeier P. , Haubitz B. , Gottlieb J. , Mattner L. , Strueber Martin
Published: March 2005

Von Jan 2002 bis Dez 2003 wurden alle Lungentransplantierten der Medizinischen Hochschule Hannover prospektiv während des postoperativen Krankenhausaufenthalts hinsichtlich der Entwicklung von invasiven Fadenpilzmykosen beobachtet. Patienten wurden als positiv eingestuft, wenn die EORTC‐Kriterien ‘probable or proven’ erfüllt waren. Retrospektiv wurde ermittelt, welche antimykotische Prophylaxe die Patienten erhielten. Von 157 lungentransplantierten Patienten entwickelten 8 eine invasive Mykose (Inzidenz 5.1% nach 17 ± 10 Tagen postoperativ). Sie führten zu einer 14‐fach erhöhten Mortalität (OR 13.8, CI95% 2.5–82, P = 0.001). Präoperative Kolonisierung der Atemwege mit Aspergillus stellte einen signifikanten Risikofaktor dar (P < 0.001, OR 21.9, CI95% 4.9–97). 101 Patienten erhielten vom ersten postoperativen Tag an Itraconazol als antimykotische Prophylaxe. 6 von ihnen entwickelten eine invasive Aspergillose (4.7%). 38 Patienten erhielten eine erst nach >14 Tagen einsetzende antimykotische Prophylaxe mit Itraconazol. Von diesen entwickelten 2 Patienten (3%) eine invasive Aspergillose. Bei 18 Patienten, von denen 10 präoperativ mit Aspergillus besiedelt waren, wurde seit Ende 2002 eine Prophylaxe mit Voriconazol in den ersten 30 postoperativen Tagen durchgeführt. Darunter kam es zu einer Zygomykose. Invasive Fadenpilzmykosen treten bei Lungentransplantierten trotz einer antimykotischen Prophylaxe mit einer hohen Inzidenz von 5% in der frühen postoperativen Phase auf und führen zu einer hohen Mortalität. Ein Management, bei definierten Hochrisikopatienten eine Voriconazol‐Prophylaxe durchzuführen, scheint einer Itraconazolprophylaxe überlegen zu sein. Zur definitiven Klärung sind jedoch noch kontrollierte Studien an größeren Patientenkollektiven erforderlich.
Antimykotika unter kontinuierlicher Nierenersatztherapie
Authors: Muhl Elke
Published: March 2005

Bei kritisch kranken Patienten mit akutem Nierenversagen und kontinuierlicher Nierenersatztherapie (CRRT) ist die Elimination von Antimykotika vielen Einflüssen unterworfen, wobei u.a. Art und Dosis des Nierenersatzverfahrens, aber auch Medikamenteneigenschaften eine Rolle spielen. Hohe Eiweißbindung, hohes Molekulargewicht, und keine oder nur geringe renale Elimination auch beim Nierengesunden sind Hinweise darauf, dass eine Substanz durch kontinuierliche Nierenersatzverfahren nicht eliminiert wird. Dies trifft für Amphotericin B, Caspofungin, Itraconazol und Voriconazol zu, nicht aber für Fluconazol. Für Fluconazol ist entsprechend der Art und Dosis der CRRT ggfs. eine Dosiserhöhung gegenüber der Dosis nierengesunder Patienten erforderlich, da die Clearance der CRRT die Clearance Nierengesunder deutlich übersteigen kann. Tagesdosen von 800 mg und mehr können nötig sein. Wegen der vielfältigen Einflüsse auf die Elimination bei Patienten unter CRRT kann auf die Bestimmung von Fluconazol‐Talspiegeln nicht verzichtet werden, um Unterdosierungen und ein Versagen der antimykotischen Therapie zu vermeiden.
Therapie der intraabdominellen Pilzinfektionen
Authors: Kujath P. , Bouchard R. , Nolde J. , Scheele J.
Published: March 2005

Candida albicans ist der am vierthäufigsten nachgewiesene Erreger bei chirurgischen Intensivpatienten. Im Verlauf schwerer Peritonitiserkrankungen muss diesen Erregern zunehmend Beachtung geschenkt werden. Für den Therapeuten ist es schwierig, die Bedeutung einer positiven Candida‐Kultur einzuschätzen. Entscheidend ist die Fage, ob es sich um eine Kontamination oder eine schwere Organmykose mit hoher Letalität handelt. Es wird deshalb versucht, die Pilzperitonitis entsprechend ihrer Ausprägung in vier Stadien einzuteilen. Aus diesen Stadien muss sich die Indikation zur antimykotischen Therapie ableiten. Stadium I ist die initiale Kontamination von Candida spp. der Peritonealhöhle nach einer Organperforation. Stadium IIa ist die Persistenz von Candida im Abdomen bei Patienten ohne Risikofaktoren, IIb Persistenz des Erregers mit Risikofaktoren. III Histologischer Nachweis einer Invasion von Candida ins Peritoneum, Stadium IV Generalisation mit Fungämie und Pilzsepsis. Eine antimykotische Therapie ist ab dem Stadium IIb indiziert.
Rezidivierende Candida‐Sepsis mit protrahiertem Lungenversagen und schwerer Leberdysfunktion
Authors: Lewejohann J. , Hansen M. , Zimmermann C. , Muhl Elke , Bruch H. P.
Published: March 2005

Systemische Pilzinfektionen, vor allem Pilzpneumonien und Pilzseptikämien werden zunehmend auf Intensivstationen beobachtet. Risikofaktoren für die Entwicklung einer Candida‐Mykose sind vor allem eine multimodale Antibiotikatherapie sowie eine Immunschwäche, entweder infolge schwerer Grunderkrankungen oder nach Transplantationen iatrogen induziert. Als häufigster Erreger wird Candida albicans nachgewiesen. Nur bei massiver Fungämie lassen sich die Erreger in der Blutkultur nachweisen.

Wir berichten über einen 50‐jährigen Patienten mit einer rezidivierenden systemischen Candida‐Mykose: Distale Ösophagusruptur nach Ballondilatation bei Cardia‐Achalasie mit Mediastinalemphysem und Mediastinitis. Schweres ARDS nach Aspiration mit septischem Schock und zunächst Nierenversagen. Langzeitbeatmung, Nierenersatztherapie und multimodale Antibiotikatherapie. Schon frühzeitig Nachweis von Candida albicans in der BAL und in mehreren Blutkulturen. Weiterhin im Verlauf auch Nachweis von Pseudomonas spp. in der BAL und Staphylokokken sowie Enterokokken in mehreren Blutkulturen. In der Folge Entwicklung einer schweren Leberdysfunktion mit den laborchemischen Zeichen einer intrahepatischen Cholestase. Anfangs auch Gerinnungsversagen und deshalb Durchführung einer Leberersatztherapie (MARS®, Gambro AB, Stockholm, Sweden) und zunächst Abfall der deutlich erhöhten Bilirubinspiegel verbunden mit einer Besserung der klinischen Situation. Im Verlauf mehrfach erneute Infektionsschübe mit Candida‐Nachweisen in mehreren bronchoalveolären Lavagen und Blutkulturen. Jeweils nach Beginn eines Infektionsschubes Anstieg der Bilirubinspiegel mit den laborchemischen Zeichen einer intrahepatischen Cholestase. Jedes Mal Besserung mittels antimykotischer Therapie, anfangs mit Voriconazol, später mit Caspofungin und Fluconazol. Zunehmende klinische und laborchemische Zeichen einer Immunschwäche im Verlauf der Intensivtherapie.

Das klinische Spektrum der Candidose ist sehr vielfältig. Systemische Candida‐Mykosen lassen sich häufig bei Patienten mit einer Immunschwäche nachweisen. Ein rezidivierende Candida‐Sepsis mit einem protrahiert verlaufenden Lungenversagen in Verbindung mit einer schweren Leberdysfunktion im Sinne einer cholestatischen Hepatose, die sich mehrmals durch eine moderne antimykotische Therapie in Kombination mit einer Intensivtherapie und Organersatztherapie bessern lässt, tritt jedoch vergleichsweise selten auf.
Pulmonale Zygomykose – eine seltene angioinvasive Pilzinfektion
Authors: Lewejohann J. , Muhl Elke , Birth M. , Kujath P. , Bruch H. P.
Published: March 2005

Rhizopus‐Arten sind Pilze, die Zygomykosen verursachen. Die Zygomykose umfasst mukokutane, rhinocerebrale, pulmonale, urogenitale und disseminierte Infektionen. Prädisponierende Faktoren sind eine Immunsuppression infolge von schweren Erkrankungen, Immundefekte oder Störungen des Stoffwechsels wie die diabetische Ketoazidose. Rhizopus‐Infektionen sind charakterisiert durch ein angioinvasives Wachstum der Pilze, eine perineurale Invasion und Nekrosen des infizierten Gewebes. Bemerkenswert ist die histopathologisch nachweisbare Invasion von Blutgefässen. Zygomykosen verlaufen häufig letal. Wir berichten über zwei Patienten mit einer pulmonalen Zygomykose, hervorgerufen durch Rhizopus spp.: Pat. 1, weibl. 73 Jahre: Protrahierter Verlauf nach infizierter Hüft‐TEP (abkurtzung für Totalendoprothese) und Infektion eines femoropoplitealen Bypasses des rechten Beines, schließlich Hüftgelenksexartikulation rechts, Pseudomonas‐Pneumonie, Staphylokokkensepsis, schweres septisches Schockgeschen mit akutem Lungen‐ und Nierenversagen, Zustand nach multimodaler Antibiotikatherapie. In der Folge Nachweis von Rhizopus spp. in der BAL, antimykotische Therapie mit Amphotericin B. Pat. 2, männl., 68 Jahre: Zustand nach Nierentransplantation, aktuell Transplantatversagen. Multiple abgelaufene Infektionen. In der Folge abszedierende Pneumonie rechts mit Entwicklung eines Pleuraempyems, darin Nachweis von Rhizopus spp., deshalb Unterlappenresektion und Transplantatnephrektomie wegen Rejektion. Protrahierter septischer Verlauf. Behandlung mit Caspofungin. Beide Verläufe endeten trotz aller intensivmedizinischer Bemühungen letal. Die seltene pulmonale Zygomykose infolge einer Infektion mit Rhizopus spp. entstand bei unseren Patienten infolge einer Immunsuppression, bei einem aufgrund einer Organtransplantation iatrogen induziert, beim anderen infolge einer protrahiert verlaufenden schweren Sepsis. Im Vergleich zu anderen Pilzinfektionen bleibt die Therapie von Rhizopus‐Infektionen schwierig. Ihre starke Affinität zu Gefäßen, in denen sie sich vermehren und ihre Eigenschaft, im Rahmen der vaskulären Invasion eine Thrombosierung und anschließende Infarktbildung herbeizuführen, erschweren die Therapie.
Pilzsepsis – Therapeutische Strategie
Authors: Staubach K. H.
Published: March 2005

Die Pilzsepsis stellt infolge der massiv angestiegenen systemischen Mykosen auch bei nicht neutropenischen Intensivpatienten ein nicht unerhebliches Problem dar. Aufgrund des großzügigen Einsatzes von Breitspektrumantibiotika erhält die physiologische Candida‐Besiedelung eine pathologische Bedeutung. Die Therapie der Pilzsepsis besteht aus kausalen, supportiven und adjunktiven Maßnahmen sowie einer so genannten Sepsisprophylaxe. Allein die sparsame und gezielte Applikation von Antibiotika könnte als Sepsisprophylaxe die Inzidenz und Letalität senken. Supportive und adjunktive Maßnahmen wie die zielorientierte Herz‐Kreislauf‐Therapie, die niedrig dosierte Kortikosteroid‐Therapie, die Applikation von aktiviertem Protein C und das Erreichen einer Normoglykämie können nach konsequenter Fokussanierung und Standardintensivtherapie das Outcome der Sepsispatienten signifikant verbessern.
SOLUBLE BIO‐MARKERS IN VASCULAR DISEASE: MUCH MORE THAN GAUGES OF DISEASE?
Authors: Woollard Kevin J
Published: March 2005

1. In recent years demonstration of a direct association between slightly elevated serum levels of soluble proteins including the acute phase response proteins, selectins and intercellular adhesion molecules and the risk of developing vascular disease have been widely reported. These studies may provide the clinician with an insight into disease diagnosis, prognosis and disease activity.

2. The simplest interpretation of this data is that soluble proteins are just sensitive markers of inflammation. However, they may in fact be modulating inflammation directly through interaction with circulating cells.

3. Recent work has shown that these soluble proteins do indeed remain active and can bind to functional ligands expressed by circulating leucocytes. The current review focuses on the soluble proteins C‐reactive protein and soluble P‐selectin and describes previous studies characterizing their interaction with immune cells to modulate the pathogenesis of vascular disease.

4. The current review focuses on the soluble proteins C‐reactive protein and soluble P‐selectin and describes previous studies characterizing their interaction with immune cells to modulate the pathogenesis of vascular disease.