Not another 4th of July report: uncommon blast injuries to the hand.

OBJECTIVES: Blast injuries to the hand are rare during peacetime and are mainly caused by fireworks. The injury patterns combine a variety of tissue destruction (laceration, dissemination, avulsion, blast, crush and burns). Emergency department staff play a key role in identifying the cause of injury, recognising the full extent of the lesion and referring patients for appropriate treatment. A review was undertaken to examine specificities in emergency department diagnosis and treatment of a separate subgroup of blast injuries. METHODS: The diagnosis and treatment of patients admitted with work-related blast injuries of the hand were retrospectively reviewed. Demographic, clinical and diagnostic data were evaluated and treatment algorithms were analysed. RESULTS: Treatment algorithms of 14 patients suffering blast injuries of the hand due to a vole captive bolt device were analysed. The non-homogeneous injury pattern showed complex multistructural lesions. Relatively innocent-looking superficial wounds mask extensive deep tissue damage, the full extent of which could only be recognised after rigorous surgical exploration. All patients but one were treated by immediate surgery, debridement of tissue necrosis and lavage. A delay before surgery resulted in phlegmonous infection in one case. CONCLUSION: Emergency staff must be aware of the potential dangers of this subgroup of blast injuries and the worsening effect of delay before surgery. Only knowledge of the underlying mechanism of the accident enables the emergency physician to understand the complexity and full extent of the injury pattern and to refer patients early for appropriate surgical management. Conservative treatment is inappropriate, dangerous and may become a focus of negligence claims.

Devastating femoral osteomyelitis after anterior cruciate ligament reconstruction.

Osteomyelitis following arthroscopic assisted anterior cruciate ligament (ACL) reconstruction has not been reported in literature. We describe an aggressive progression of septic arthritis and osteomyelitis leading to the complete destruction of the condylar region in a young non-immunosuppressed patient after reconstruction of the ACL. In addition we discuss the steps in diagnostics and our salvage procedures.

Oldie but goldie: Bristow-Latarjet procedure for anterior shoulder instability.

PURPOSE: To analyse the functional and radiological outcomes of the Bristow-Latarjet procedure in patients with recurrent anterior glenohumeral instability. METHODS: Records of 29 patients were reviewed retrospectively. Date of first dislocation, injury mechanism, and number of recurring dislocations before and after surgery were recorded. The overall function and stability of the shoulder was evaluated. RESULTS: 24 (83%) of the glenohumeral instabilities were caused by trauma. The mean number of recurring dislocations was 8 (95% confidence interval [CI], 0-18); one patient had had 40 recurrences. No dislocation ensued postoperatively. The overall functional outcome was good, with a mean Rowe score of 90 points (95% CI, 78-100). Scores of 17 (59%) of the patients were excellent, 7 (24%) were good, 3 (10%) were fair, and 2 (7%) were poor. CONCLUSION: The Bristow-Latarjet procedure is a good surgical treatment for recurrent anterior-inferior instability of the glenohumeral joint.

[Results after treatment of instable fractures of the proximal humerus using a fixed-angle plate]. / Ergebnisse nach Versorgung instabiler proximaler Humerusfrakturen mittels winkelstabiler Platte.

BACKGROUND: Fixed-angle implants are being increasingly used in surgery of fractures of the proximal humerus. The aim of this retrospective investigation was to evaluate the outcome after fracture reduction utilizing a fixed-angle plate (Königsee). MATERIALS AND METHODS: Between January 2003 and April 2004, 58 patients were operated, 52 of whom received a fixed-angle implant; 46 cases were harvested for a follow-up examination. Each patient was re examined clinically and radiologically at least 6 and 18 months after surgery. The functional outcome was evaluated using the Constant Score and the Simple Shoulder Test. The results were compared to results of other investigations. RESULTS: The mean patient age was 68.8 years (34-94 years). Fractures were classified using the Neer Classification: 12 were classified as two-part, 25 as three-part, and 9 as four-part fractures. Three of the three-part and four of the four-part fractures were rated as luxation fractures. The overall functional outcome of all cases was good. More than 18 months after surgery the mean general "Constant Score" was 57; the mean side-related "Constant Score" was 89%. The Simple Shoulder Test revealed a pain-free range of motion in 41 (89.1%) of the individuals. The majority of the patients were satisfied with the results regarding remaining range of activity of the injured limb. In five cases significant complications occurred. In two cases the head of the humerus collapsed, and in one case a necrosis of the head occurred. In one individual the implant broke after an additional trauma. In this case a re-osteosynthesis utilizing a tibia plate was performed and the patient was excluded from further follow-up investigations. One soft tissue infection occurred after initial surgery. CONCLUSION: It has been shown that results after fixation of proximal humerus fractures with fixed-angle implants are good. The functional outcome is good and complications are rare. Our results correlate with other investigations regarding fracture reduction using fixed-angle plates and nails.

Does the frontal airbag avoid thoracic injury?

INTRODUCTION: The airbag is an established car safety device. However, recent studies pointed out that even the airbag might cause injuries. Nevertheless, most physicians do consider a lower risk in accident victims sustaining severe injury of the chest, when a deployed frontal airbag has been reported. We set out to verify the frequency and pattern of thoracic injury in car drivers protected by a frontal airbag during traffic accidents. METHODS: This investigation was conducted as part of a prospective surveillance analyzing traffic accidents. Enrolled were car drivers included in a databank between January 2001 and December 2004 consecutively. The chance for sustaining chest injury with or without a frontal airbag was described using the relative risk. RESULTS: A total of 188 car drivers were included in the analysis. In 54 (28.7%) cases a deployed airbag and in 134 (71.3%) the absence of an airbag has been documented. Out of those cases 16 (29.6%) drivers with airbag and 30 (22.4%) without airbag sustained a chest injury. The mean abbreviated injury scale (AIS) of chest injuries in drivers with deployed airbag was 2.3 (1-5; SD +/- 1.45; mean injury severity scale [ISS] 21.1 [SD +/- 17.18]), in drivers without airbag 1.6 (1-4; SD +/- 1.12; mean ISS 15.8 [SD +/- 20.6]). For belted drivers with an airbag the relative risk to sustain chest injury was 1.96 compared to those without an airbag. CONCLUSIONS: The airbag does not avoid chest injury definitively. Much more, it has been demonstrated that the relative risk to sustain relevant thoracic injury seems to be almost higher in restrained drivers with a frontal airbag.

Failed or inadequate bone marrow aspiration: a fast, simple and cost-effective method to produce a cell suspension from a core biopsy specimen.

Failure to aspirate bone marrow (BM) diminishes diagnostic accuracy and efficiency because BM cell suspensions are crucial for modern haematological diagnostic methods such as cytomorphology, flow cytometric immunophenotyping (FCI), cytogenetics or fluorescent in situ hybridization (FISH). We mechanically disaggregated unfixed BM core biopsies with the Dako Medimachine in 65 cases of macroscopically suspected dry taps. Cytospins, three-colour FCI and in some cases karyotyping and FISH were performed successfully. Most cytospins (34 of 50; 68.0%) were of good quality, while a further 18.0% showed moderate but still informative quality. FCI showed good quality in 36 of 60 (60.0%) cases; in 13.3% quality was moderate, but diagnostically useful results were obtained. Surprisingly, all four cases of formerly undiagnosed BM-carcinosis could be clearly detected on cytospins. Finally, five of seven (71.4%) attempts yielded analysable metaphases mostly in cases where no metaphases could be obtained from BM or peripheral blood. The described method of mechanical disaggregation of unfixed BM core biopsies compares favourably with other published approaches, allowing the application of all techniques where BM cell suspensions are needed. Thus, it can help to establish the underlying diagnosis in patients with abnormalities in peripheral blood and unsuccessful marrow aspirations.

CD38 expression is an important prognostic marker in chronic lymphocytic leukaemia.

Employing a multicolour flow cytometry assay, 133 B-chronic lymphocytic leukaemia (B-CLL) cases were analysed for surface expression of CD38. Based on a cut-off value of 20%, CLL patients were categorised into a CD38-positive (> or = 20%, n = 56) and a CD38-negative subgroup (< 20%, n = 77) and separately analysed for clinical and laboratory parameters. Patients in the CD38-positive cohort were characterised by an unfavourable clinical course with a more advanced disease stage, poor responsiveness to chemotherapy, short time to initiation of first treatment and shorter survival. In contrast, the CD38- negative group required minimal or no treatment, remained treatment-free for a longer time period and had prolonged survival (P < 0.05). CD38 expression was a robust marker in the majority of patients in that it was stable over time and not significantly influenced by chemotherapy. In conclusion, our data confirm recent studies suggesting a role of CD38 as a predictor of clinical outcome in patients with B-CLL.

The histone acetyltransferase activity of PCAF cooperates with the brahma/SWI2-related protein BRG-1 in the activation of the enhancer A of the MHC class I promoter.

Major histocompatibility complex (MHC) class I proteins are an essential component of the immune system allowing the organism to protect from viral infections and neoplastic transformation. Expression of the MHC class I genes is regulated by a variety of cis-regulatory promoter elements among which the enhancer A is of particular importance. This enhancer is synergistically activated through AP-1/ATF and NF-kappa B transcription factors. NF-kappa B recruits the histone acetyltransferase (HAT) p300/CREB-binding protein (CBP) to the multiprotein complex bound to the enhancer A. Here we present evidence that acetylation and deacetylation processes are involved in the activation of the enhancer A. The p300/CBP associated factor PCAF, but not p300/CBP, counteracts the repression of the enhancer A mediated by the histone deacetylase HDAC1. Furthermore, overexpression of PCAF results in an increase in the acetylation of histone H4 bound to the enhancer A and HDAC1 counteracts the PCAF-mediated H4 acetylation. The activation function of PCAF requires the p300/CBP binding motif indicating that PCAF might be recruited to the enhancer A through an association with p300/CBP. Moreover, PCAF and the Brahma/SWI2-related protein BRG-1, which is a key factor of the human ATP-dependent chromatin remodelling complex SWI/SNF, synergistically up-regulate the enhancer A. Synergistic activation requires the HAT domain of PCAF. Taken together our data suggest that members of two different groups of chromatin modifying complexes are involved in the activation of the enhancer A of the MHC class I promoter.

Differential expression of chemokine receptors in B cell malignancies.

Chemokines are a family of 8-10 kDa proteins with a wide range of biological activities including the regulation of leukocyte trafficking, modulation of haemopoietic cell proliferation and adhesion to extracellular matrix molecules. Using a panel of chemokine receptor-specific monoclonal antibodies (MoAb) in a multicolour flow cytometry approach we analysed the expression of the lymphocyte-associated chemokine receptors CXCR4, CXCR5, CCR5 and CCR6 in B cell acute lymphoblastic leukaemia (precursor B-ALL; six cases), B cell chronic lymphocytic leukaemia (B-CLL; 31 cases), multiple myeloma (10 cases), mantle cell lymphoma (MCL, four cases), follicular lymphoma (FL, three cases) and hairy cell leukaemia (HCL, five cases). We demonstrate that CXCR4, CXCR5 and CCR6 are differentially expressed in these B lymphoproliferative disorders depending on the maturational stage of the malignant B cell population investigated. In particular, we found that CXCR4 is strongly expressed on immature ALL blasts whereas no surface immunoreactivity for CXCR5, CCR5 and CCR6 was observed. By contrast, non-Hodgkin's lymphomas (NHLs) corresponding to more mature peripheral B cell subsets (ie B-CLL and MCL) exhibited high expression levels of CXCR4 and CXCR5. Analysis of terminally differentiated myeloma cells revealed a down-regulation of CXCR4, CXCR5 and CCR6. CCR5, which is not expressed in normal B cells, was also absent from the majority of NHLs. However, CCR5 staining was seen in three of five cases of HCL, representing the first example of cross-lineage aberrant chemokine receptor expression in malignant haemopoietic cells.

A multiprotein complex consisting of the cellular coactivator p300, AP-1/ATF, as well as NF-kappaB is responsible for the activation of the mouse major histocompatibility class I (H-2K(b)) enhancer A.

Major histocompatibility complex (MHC) class I genes encode highly polymorphic antigens that play an essential role in a number of immunological processes. Their expression is activated in response to a variety of signals and is mediated through several promoter elements among which the enhancer A is one of the key control regions. It contains binding sites for several transcription factors, for example: (i) a well-characterized binding site for rel/NF-kappaB transcription factors in its 3'-end (the H2TF1 or kappaB1 element), (ii) a second kappaB site (the kappaB2 element), which is located immediately adjacent 5' to the H2TF1 element and which is recognized by p65/relA in the human HLA system, and (iii) an AP-1/ATF recognition sequence in the 5' end (EnA-TRE). Here we demonstrate that latter element is bound by at least two distinct heterodimers of the AP-1/ATF transcription factor family, namely c-Jun/ATF-2 and c-Jun/Fra2. Moreover, our data reveal that the enhancer A is simultaneously bound by AP-1/ATF and rel/NF-kappaB transcription factors and that the cellular coactivator p300, which enhances enhancer A-driven reporter gene expression if cotransfected, is recruited to the enhancer A through this multiprotein complex. In contrast to the complete enhancer A, neither the EnA-TRE nor the H2TF1 element on their own are able to confer activation on a heterologous promoter in response to the phorbol ester tumor promoter TPA or the cytokine TNFalpha. Moreover, deletion of any one of the enhancer A control elements results in a dramatic loss of its inducibility by TNFalpha, and point mutations in either the EnA-TRE or the H2TF1 element lead to the loss of AP-1/ATF or NF-kappaB binding, respectively, and to the loss of enhancer A inducibility. Therefore, we conclude that the enhancer A is synergistically activated through a multiprotein complex containing AP-1/ATF, NF-kappaB transcription factors as well as the cellular coactivator p300.

Comparative flow cytometric study of clonal excess in leukaemic peripheral blood from patients suffering from chronic lymphocytic leukaemia (B-CLL) by different antibodies, staining techniques and the effects of blood storage.

This investigation studied the effects of cell preparation methods, different antibody panels and blood storage on antigen expression of abnormal B lymphocytes from patients with B-CLL. Blood specimens collected in Heparin de novo were processed by using conventional Hypaque-Ficoll density gradient centrifugation and whole blood lysis. These were stored for 3 days at 4 degrees C, 24 degrees C and 30 degrees C. Although clonal excess was detected by all antibody panels, significant differences could be observed in terms of molecules of equivalent fluorochromes (MEF/MESF units). Evaluation of 'weak and strong' staining is dependent on the antibody panel used. Immunofluorescent values for CD19 and CD45 were unchanged at 4 degrees C and 24 degrees C but immunoglobulin staining showed best results when blood was stored at 4 degrees C. Storage at 30 degrees C produced unreliable results. Abnormal B lymphocytes should be analysed immediately after the specimen is obtained. If shipment is necessary they should be kept at 4 degrees C. Surface immunoglobulins are the 'antigens' most sensitive to storage alterations. Sample alterations alone are sufficient to the correct classification of NHL, especially in the case of low-grade NHL.

Multiparametric immunophenotyping of B cells in peripheral blood of healthy adults by flow cytometry.

The investigation of patients suffering from malignant lymphomas of the B-cell type requires flow cytometric immunophenotyping. Several reports described the expression of almost all B lineage antigens on normal and abnormal B lymphocytes. Thus, immunophenotyping of lymphomas must be interpreted in the context of the reference values obtained for healthy control individuals. For this purpose multiparametric flow cytometric analysis offers the unique feature for lymphocyte subset analysis. In the present study B lymphocytes in the peripheral blood of healthy adults were investigated by multiparametric flow cytometric immunophenotyping for the detection of the frequency (in percent) of antigens provided by the revised European-American classification of lymphoid neoplasms (REAL) classification. Thus, 84 healthy adults were investigated and grouped by age (average ages were as follows: group 1, 25.38 years; group 2, 33.86 years; group 3, 44.17 years; group 4, 55.67 years; group 5, 66.67 years). Analysis was done for surface immunoglobulins (kappa and lambda chains of immunoglobulin M [IgM] and IgD) as well as CD10, CD11c, CD23, CD38, CD103, FMC-7, and B-B4. Three-color immunophenotyping was performed for kappa/CD19/CD5, lambda/CD19/CD5, surface IgM/surface IgD/CD19, FMC-7/CD19/CD5, CD103/CD11c/CD19, CD10/CD23/CD19, and CD38/B-B4/CD19 by live gating of CD19+ events (n = 2,000). Although some numerical differences could be obtained for the different groups, statistical differences (P < 0.005) could only be obtained for the CD19+/CD5+ B-cell subset, which was decreased in the elderly patients (group 5). The established two-color and three-color stainings will serve as a basis for future multiparametric immunophenotyping of abnormal lymphocytes (e.g., for patients suffering from non-Hodgkin's lymphoma of the B-cell type).

Lymphocyte gating of peripheral blood in patients with leukemic low-grade non-Hodgkin's lymphoma by multiparametric flow cytometry.

The standardized fluorescence intensity as expressed in molecules of equivalent soluble fluorescence (MESF) of lymphocytes from normal individuals and patients suffering from low-grade non-Hodgkin's-lymphomas was obtained comparing different staining patterns of CD45(FITC) and CD20(PerCP). After standardization of the flow cytometer using standardized fluorescent particles ('beads') significant differences could be obtained for hairy cell leukemia, chronic lymphocytic leukemia and immunocytomas in the peripheral blood. In contrast, centroblastic-centrocytic as well as centrocytic lymphomas showed no significant variations as compared to normal peripheral blood lymphocytes. According to these results, a new lymphocyte gating procedure was established by adding CD14(PE) and three-color measurement by CD45/CD14/CD20 staining of peripheral blood using erythrocyte lysis. The established gating procedure leads to a crucial discrimination and quantification of abnormal and normal lymphocytes per one measurement, whereas the 'leucogate' as defined by CD45/CD14 staining alone was insufficient for correct lymphocyte gate setting. In conclusion, the different staining of CD45 and CD20 in leukemic peripheral blood should be considered when fluorescence intensity or atypical peaks occurred in flow cytometric histograms suggesting for abnormal cell populations. In addition, it is possible to use this information to classify low-grade lymphomas.

Immunophenotyping of B lymphocytes by multiparametric flow cytometry in bone marrow aspirates and peripheral blood of healthy adults.

Establishing reference ranges by multiparametric immunophenotyping of mature B cells in bone marrow and peripheral blood of healthy adults is of interest because the detection of bone marrow infiltration, persistance of light chain restriction as well as discrimination between reactice and malignant lymphocytes are important applications of B-cell immunophenotyping. To determine the pattern of antigens as expressed by malignant mature B lymphocytes, bone marrow aspirates and peripheral blood of healthy adults in the present study were investigated for the presence and percentage frequency of those antigens as defined for immunophenotyping of B-cells by the REAL-Classification. For this purpose analysis of CD19 positive B lymphocytes by Live Gate analysis was performed. The established two-color as well as three-color stainings will serve as a basis for future investigations designed to test multiparametric analysis of B lymphocytes in bone marrow aspirates and peripheral blood. In conclusion, all investigated antibodies stained in varying percentage frequency on B-cell subtypes and statistical significant differences could be considered only for the CD19/CD10 staining in bone marrow aspirates. On the basis of this analysis, all the reported lineage antigen combinations are present both in malignant B lymphocytes as well as normal B cells in considerable percentage frequency. These findings are of important interest for follow-up investigations of patients with non-Hodgkin s lymphomas by multiparametric immunophenotyping.

Immunophenotyping of B lymphocytes by multiparametric flow cytometry in bone marrow aspirates of healthy adults.

Establishing reference ranges by multiparametric immunophenotyping of mature B cells in bone marrow of healthy adults is of interest because the detection of bone marrow infiltration and persistence of light chain restriction, as well as discrimination between reactive and malignant lymphocytes are important applications of B-cell immunophenotyping. To determine the pattern of antigens as expressed by malignant mature B lymphocytes in the present study, bone marrow aspirates of healthy adults were investigated for the presence and percentage frequency of those antigens as defined for immunophenotyping of B cells by the REAL Classification. For this purpose, analysis of CD19-positive B lymphocytes by 'live gate' analysis was performed. The established two-color as well as three-color stainings will serve as a basis for future investigations designed to test multiparametric analysis of B lymphocytes in bone marrow aspirates. All investigated antibodies stained with varying percentage frequency on B-cell subtypes, and no statistical significant difference was found between bone marrow aspirates of women and those of men. On the basis of this analysis, all the reported lineage antigen combinations are present both in malignant B lymphocytes and in normal B cells in considerable percentage frequency. These findings are of importance for follow-up investigations of patients with non-Hodgkin's lymphomas by multiparametric immunophenotyping.