A Belgian student with black eschars.

BACKGROUND: Human cowpox virus infection is a rare zoonotic disease. Cowpox virus is a member of the Orthopoxvirus genus, like smallpox. Over the last years records of cowpox virus transmission from pet cats and pet rats to humans in Europe have increased. This observation may result from the loss of cross-immunity against orthopoxviruses after discontinuation of routine smallpox vaccination in the 1980s. CASE PRESENTATION: We report the first case of a human cowpox infection in an unvaccinated Belgian citizen. This 19-year-old student presented with multiple necrotic skin lesions on the chin, the scalp and the pubic region, and with cervical lymphadenopathy and flu-like symptoms. The diagnosis of human cowpox was based on electron microscopic findings and PCR examination performed on a skin biopsy of the pubic lesion. Close contact with cats (her domestic cats or cats from a local shelter) was probably the source of transmission. Spreading of the lesions was likely the result of autoinoculation. After six months all lesions spontaneously healed with atrophic scars. DISCUSSION: To enhance awareness of this rare viral zoonosis and to verify the suspected increase in incidence and symptom severity after cessation of smallpox vaccination, one could argue whether human cowpox should become a notifiable disease.

Biological tests carried out on serum/plasma samples from donors of human body material for transplantation: Belgian experience and practical recommendations.

This paper on the biological tests carried out on serum/plasma samples from donors of human body material (HBM) is the result of a project of the working Group of Superior Health Council of Belgium formed with experts in the field of HBM and infectious serology. Indeed, uncertainty about the interpretation of biological test results currently leads to the sometimes unjustified cancelling of planned donations or the rejection of harvested HBM, whilst more sophisticated diagnostic algorithms would still allow the use of organs or HBM that would otherwise have been rejected. NAT tests will not be discussed in this publication. In the first part some general aspects as the need for a formal agreement between the Tissue Establishment l and the laboratory responsible for the biological testing, but also some specifications regarding testing material, the choice of additional biological tests, and some general aspects concerning interpretation and reporting are discussed. In a second part, detailed information and recommendations concerning the interpretation are presented for each of the mandatory tests (human immunodeficiency virus, hepatitis B virus, hepatitis C virus and syphilis) is presented. A number of not mandatory, but regularly used optional serological tests (e.g. for the detection of antibodies to Toxoplasma gondii, Epstein-Barr virus, human T cell leukemia virus and cytomegalovirus) are also extensively discussed. Although the project was meant to provide clarification and recommendations concerning the Belgian legislation, the majority of recommendations are also applicable to testing of donors of tissues and cells in other (European) countries.

The risk of transmitting cutaneous malignancy through skin transplantation: a literature-based risk assessment.

According to the European Union Tissues and Cells Directives donation of tissue is contraindicated in the presence of or a previous history of malignant disease, with the exception of cutaneous basal cell carcinoma. Skin cancer is the most common cancer. Due to ultraviolet light exposure and increasing life expectancy an increasing prevalence of malignant or premalignant skin lesions is observed, which may result in a decline of the availability of skin for transplantation. A risk assessment based on published studies and expert opinion was performed in order to investigate the risk of transmitting malignant or premalignant skin lesions through tissue transplantation, and more particular through skin transplantation. The scarcity of data concerning cancer transmission in tissue transplantation was challenging. Circumstantial evidence, available for organ transplantation, was used to develop the following policy proposal for skin transplantation and cutaneous tumours. Malignant melanoma is an absolute contraindication for the donation of skin and also of other tissues, whereas, non-lesional skin and other tissues of a donor with non-melanoma skin cancer (basal cell and squamous cell carcinoma) or with a premalignant skin lesion can be considered for transplantation. The above mentioned protocol proposal might serve as a prototype for analogous protocols for non-cutaneous malignancies.

Report of the clinical donor case workshop of the European Association of Tissue Banks annual meeting 2012.

The European Association of Tissue Banks (EATB) donor case workshop is a forum held within the program of the EATB annual congress. The workshop offers an opportunity to discuss and evaluate approaches taken to challenging situations regarding donor selection, it promotes consensus development in deciding tissue donor acceptability when donor health issues are not addressed in standards and regulations, and serves to strengthen the professional tissue banking networks across Europe and beyond. This report reflects some of the discussion at the workshop during the annual congress in Vienna in 2012. The cases presented dealt with problems encountered by tissue bank facilities concerning idiopathic thrombocytopenia and auto-immune disorders, hemodilution and blood sample identification, premalignant and malignant lesions, and Huntington's disease. The discussions during the workshop demonstrate that the implications on the safety of tissue transplantation of various tissue donor illnesses, physical findings and behaviours, and the preventive measures taken by tissue facilities, may not always be agreed by tissue facility medical directors and other professionals. Moreover, they reveal that operating procedures, regulations and standards cannot comprehensively cover all tissue donor findings, medical histories and circumstances surrounding the cause of death. For many of the issues raised, there is a need for scientific research to provide a better evidence base for future deliberations about the suitability and eligibility of tissue allograft donors.

The 22nd annual meeting of the European Tissue Repair Society (ETRS) in Athens, Greece.

The 22nd Annual Meeting of the European Tissue Repair Society, Athens, Greece, October 4 to 5, 2012 informed about pathophysiological mechanisms in tissue repair and on the development of clinical treatments of chronic wounds, fibrosis, and cancer, considering recent advances in molecular biology and biotechnology.

Keratinocytes in the treatment of severe burn injury: an update.

Burns are among the most life-threatening physical injuries, in which fast wound closure is crucial. The surgical burn care has evolved considerably throughout the past decennia resulting in a shift of therapeutic goals. Therapies aiming to provide coverage of the burn have been replaced by treatments that have both functional as aesthetic outcomes. The standard in treating severe burns is still early excision followed by skin grafting. The use of cultured keratinocytes to cover extensive burn wounds appeared very promising at first, but the technique still has several limitations of which the long time to culture, the major costs, the risk of infection and the need for an adequate dermal layer limit clinical application. The introduction of dermal substitutes, composite grafts, tissue engineering based on stem cell application have been advocated. The aim of this review is to assess the use of cultured keratinocytes in terms of technical aspects, clinical application, limitations and future perspectives. Cultured keratinocytes are expected to keep playing a role in wound healing, especially in the field of chronic wounds. In severe burns, despite its limitations, keratinocytes can be beneficial if implemented as one of the elements in a broader wound management.

Fifteen years of infrapopliteal arterial reconstructions with cryopreserved venous allografts for limb salvage.

OBJECTIVE: The availability of autologous vein grafts remains the limiting factor in infragenual bypass surgery in many patients with critical limb ischemia (CLI). Alternatives such as prosthetic conduits are known to have a poor outcome and most are not resistant to infection. Based on previous experimental work, we started to use cryopreserved saphenous vein allografts for this indication 15 years ago. To evaluate their outcome, we performed a retrospective study of those bypasses with distal anastomosis on a crural or pedal vessel. METHODS: Between November 1991 and December 2005, 108 cryopreserved great saphenous vein allografts were implanted for in 92 patients (42 women, 50 men) with a mean age of 71 years (range, 39-88 years). All patients received low-dose immunosuppressive therapy for up to 1 year after intervention. Follow-up was conducted until amputation, death, or the end of the study in March 2007. RESULTS: Nondiabetic atherosclerosis the cause of CLI in 57%, 41% had diabetes, and 2% had Buerger disease. Forty-one conduits were performed for primary reconstructions and 67 for redo reconstructions. During a mean follow-up of 26.4 months, 69 occlusions occurred. Primary and secondary patency rates were 56% and 73% at 1 year, 32% and 60% at 3 years, and 17% and 38.5% at 5 years. Survival rates were 87.4% at 1 year and 64.5% at 5 years. At multivariate analysis, the intake of statins was predictive for improved patency (hazard ratio [HR], 0.09; P = .024) and for prolonged survival (HR, 0.335; P = .045). The presence of diabetes showed a trend for inferior patency (HR, 2.325; P = .116) and for decreased amputation risk (HR, 0.592; P = .078). CONCLUSION: Cryopreserved saphenous vein allografts are a valuable alternative to prosthetic materials when autologous veins are not available. Our limb salvage and patency rates are higher then those described for prosthetic grafts at the infrapopliteal level in most studies. Moreover, these grafts are resistant to infection when performed for revascularization in patients with an infected ulcer. Better graft and patient selection, better graft surveillance and immunologic matching, and standard use of statins could possibly improve the results even further. Shortage in availability might be a limiting factor for their widespread use.

How evidence-based is venous leg ulcer care? A survey in community settings.

AIM: This paper is a report of a study to describe venous leg ulcer care regarding compression, pain management and lifestyle advice in community settings and to identify factors that predict the provision of lifestyle advice by nurses. BACKGROUND: Incongruence between evidence and practice in leg ulcer care has been reported. Little is known about predictive factors related to the provision of lifestyle advice. METHOD: Two focus interviews and a Delphi procedure were used to develop a self-administered questionnaire based on the Graham questionnaire. Nurses employed by community healthcare organizations and independent nurses in private practices participated (n = 789). The data were collected in 2006. FINDINGS: Compression was applied in 58.7% of patients with venous ulcers. Pain was present in 82.9%. A third of patients with pain received analgesics, but half of these patients (52.1%) took analgesics as prescribed. Half of the nurses (50.8%) gave lifestyle advice related to the leg ulcer. It was mainly instructions about leg elevation (68.3%), promoting physical activity (39.8%) and optimizing nutrition (16.7%) that were provided. Nurses who perceived themselves to have adequate leg ulcer knowledge and skills were 3.75 times more likely to provide lifestyle advice compared with those lacking such knowledge and skills. Nurses who found leg ulcer care not rewarding, rarely successful or difficult gave statistically significantly less lifestyle advice than those who found it rather rewarding, successful and not difficult. CONCLUSION: Patients with leg ulcers receive less than optimum care and patient education. A particular challenge lies in leg ulcer education programmes and pain management.

Physical examination of the potential tissue donor, what does literature tell us?

European Tissue Banks should carry out a physical examination as a part of the donor selection procedure. This is one of the obligations concerning donation and procurement mentioned in the European Commission directives on tissue banking. As the directives do not give any further specification on the content or on the procedure of the physical examination, a search of literature was done in order to find more information. Although data in literature generally remain quite vague, it was possible to set up a list of items which should be looked at during physical examination. This list can be used temporarily until further information is gathered from an international survey and from a risk assessment analysis.

Leg ulcers: a new symptom of Blau syndrome?

Blau syndrome is a rare autosomal dominant condition, typically defined by granulomatous polyarthritis, uveitis and skin eruption. Biopsy specimens demonstrate non-caseating granulomas in all lesions. We present a case of Blau syndrome associated with large recalcitrant leg ulcers. Biopsies taken in the leg ulcers of our patient systematically showed granulomas. Although leg ulcers have not previously been described as a part of Blau syndrome, we assume that the ulcerations in this case form part of Blau syndrome.

Eye bank issues: II. Preservation techniques: warm versus cold storage.

Most of the tissue used for penetrating keratoplasty is issued through eye banks that store the corneoscleral button either in hypothermic storage at 2-6 degrees C or in organ culture at 31-37 degrees C. These two preservation techniques differ in technical aspects, tissue evaluation possibilities, storage time and microbiological safety. Hypothermic storage is simple and requires little expensive equipment. In general a pre-storage evaluation of the endothelium is performed by specular microscopy and storage time is usually around 7-10 days. Organ culture is a relatively complicated technique requiring more expertise and well-equipped facilities. Evaluation of the endothelium is not only performed before storage, but is routinely performed after storage through the use of light microscopy. With organ culture the allowed storage period is longer, up to four weeks. The vulnerability of organ culture to microbial contamination can be turned into an advantage because it allows the detection of residual micro-organisms on the cornea before surgery. Both preservation techniques seem to result in similar graft survival. The method of choice for preservation of the donor cornea is dictated by a number of factors mentioned in this review and this helps to explain the geographical differences in the use of the different techniques.

Role of natural killer cells in the rejection process of corneal allografts in rats.

BACKGROUND: The exact mechanism of human corneal allograft rejection, which is the major cause of corneal transplant failure, remains unclear. We investigated the role of natural killer (NK) cells in rat corneal allograft rejection by examining the aqueous humor (AH) cell infiltrate on different postoperative days. METHODS: Flow cytometric analysis was performed on the AH and submandibular draining lymph node (DLN) cells before transplantation and at different time points thereafter. In addition, we performed functional cytotoxicity assays with cells present in the AH during corneal rejection. RESULTS: We demonstrated a gradual increase in the absolute cell number of different hematopoietic subpopulations in the AH after allogeneic cornea transplantation. CD3CD4 cells, mainly monocytes and macrophages, were the predominant subpopulation 2 days after transplantation, followed by a successive relative increase of CD4 T cells, CD8 T cells, CD161 T cells, and NK cells. NK and CD161 T cells were present at a 10- to 15-fold higher percentage than in the DLN, suggestive of local expansion of these cells. A higher percentage of NK cells were CD8-negative compared with DLN NK cells. AH cells specifically lysed allogeneic cells, and this cytotoxicity was mainly attributable to NK cells but not to CD4 or CD8 T lymphocytes. CONCLUSION: These results confirm the crucial role of CD4 cells in the allogeneic corneal graft rejection process and implicate NK cells as possible mediators of the rejection.

Therapeutic penetrating keratoplasty: clinical outcome and evolution of endothelial cell density.

PURPOSE: To identify the influence of the underlying etiology on the outcome of therapeutic penetrating keratoplasty (PKP) in the management of corneal perforation, impending perforation or refractory corneal disease. METHODS: A retrospective observational case series with 28 patients (28 eyes) was performed. Patients were divided in two subgroups, based on the underlying ocular disease leading to the therapeutic PKP. Group I consisted of 15 patients (54%) with an infectious etiology, the other 13 patients (46%) with a noninfectious condition belonged to group II. Mean follow up was 23.2 months (median 21 months). RESULTS: An enucleation could be avoided in all but two cases. In group I, 12 of 15 (80%) grafts remained clear, compared with only 3 of 13 (23%) in group II (p = 0.0004). Visual acuity (VA) amelioration was achieved in 20 of 28 patients (71%). In group I the postoperative VA was significantly better than preoperatively (p = 0.002); however, in group II the improvement was only borderline significant (p = 0.05). Overall five out of ten (50%) patients with a therapeutic PKP in group I had an endothelial cell density >or= 1900 cells/mm2 at the time of their last specular microscopy. CONCLUSION: Our study demonstrates the importance of underlying etiology on the outcome of therapeutic PKP. Graft survival and postoperative visual acuity varied widely depending on the underlying ocular disease, and it was clear that therapeutic PKP has a much better prognosis in infectious conditions when compared with noninfectious conditions. We believe that these good results can justify the use of therapeutic PKP in selected cases of infectious keratitis.