Human brucellosis poses a significant public health problem in many developing countries and requires fast and accurate diagnosis. A PCR assay amplifying part of the 31-kDa Brucella abortus antigenic protein gene sequence was developed and applied to whole-blood and serum samples from 31 brucellosis patients and 45 healthy individuals. All patients except one had detectable Brucella DNA in either whole blood or serum (combined sensitivity, 97%), but the assay sensitivity was higher with serum samples (94%) than with wholeblood samples (61%). The assay specificity was excellent (100%). A confirmatory PCR assay targeting another Brucella gene region (omp-2) was also developed but lacked sensitivity. Serum is the optimal specimen for the diagnosis of brucellosis by PCR, a choice that leads to assay simplification and shortens turnaround time.Brucellosis is still an important zoonosis of both public health and economic significance in many developing countries. Half a million new cases are reported worldwide each year, but according to the World Health Organization, these numbers greatly underestimate the true incidence of human disease (22). As the clinical picture of human brucellosis is extremely variable, diagnosis can be established only by laboratory methods. Since the disease constitutes a serious infection necessitating treatment with a prolonged course of antibiotics, accuracy and short turnaround time are required for these tests (21).Blood cultures represent the "gold standard" of laboratory diagnosis. Automated systems have been reported to detect more than 95% of Brucella melitensis-positive cultures within 7 days of incubation (23). Unless this technology is not available, prolonged incubation, blind subcultures, and special growth media are no longer required (23). Ironically, however, the technology indeed is lacking in developing countries or rural areas where the disease is prevalent. In addition, due to their comparatively long doubling time, Brucella species grow slowly on primary cultures and subcultures, while their inert biochemical profiles hamper fast identification of isolates (9). Distinct disease conditions like focal, relapsing, or chronic disease and disease caused by species other than B. melitensis are characterized by low blood culture yields and pose special diagnostic problems (1, 2). Consequently, detection and identification of Brucella spp. in clinical specimens by cultures may still be a difficult task with significant delays.Several agglutination tests (Rose Bengal, Wright's tube, Wright's card, and Wright-Coombs) and indirect immunofluorescence, complement fixation, and enzyme-linked immunosorbent assays are also available for diagnosis of brucellosis (3, 14, 24). The standard, with which all other methods should be compared, is Wright's tube agglutination test (1,14). A broad range of test sensitivity, low specificity in areas of endemicity, lack of usefulness in diagnosing chronic disease and relapse, presence of cross-reacting antibodies, and lack of timeliness constitute ...

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Summary A number of growth factors are involved in clonal haematopoietic expansion and their clinical significance in patients with chronic myeloproliferative diseases requires further evaluation. Using enzyme‐linked immunosorbent assays, we analysed serum levels of interleukin (IL)‐1a, IL‐1b, IL‐2, IL‐6, the soluble IL‐2 receptor alpha (sIL‐2Ra), and thrombopoietin (TPO), in 25 individuals with myelofibrosis with myeloid metaplasia (MMM), 40 with essential thrombocythaemia (ET), eight with polycythaemia vera (PV), 10 patients with chronic myeloid leukaemia (CML) and 27 normal controls. These were correlated with clinicopathological characteristics including overall survival, and histopathological bone marrow features, including angiogenesis. The serum derived from patients with MMM, ET, PV and CML contained significantly higher IL‐2 and sIL‐2Ra than healthy subjects, while IL‐6 levels were higher only in MMM and CML than controls. IL‐2, sIL‐2Ra and IL‐6 levels were raised during the transformation phase of CML, during progression of MMM to AML, and ET and PV to myelofibrosis (P < 0·001). There was a positive correlation between IL‐2, sIL‐2Ra, IL‐6 and angiogenesis in bone marrow samples. Cytokines may be useful markers for predicting clinical evolution, reflecting increased angiogenesis. This requires further evaluation to guide diagnostic and therapeutic options.

The frequency of ischaemic heart disease observed after splenectomy for trauma and the low cholesterol levels found in patients with hypersplenism are observations that suggest a possible role for the spleen in lipid metabolism and in the aetiology of atherosclerosis. The present study was designed to examine this role in experimental animals. Serum levels of total cholesterol, triglyceride and high-density lipoprotein (HDL) cholesterol were determined in 32 rats. The spleen was removed in 16 rats and the remaining 16 were sham operated. Half of the splenectomized and half of the remaining rats were fed on a diet rich in fat and the two other subgroups were fed normal chow. Blood tests were performed before, and 3 and 6 months after operation. A significant increase in serum triglyceride and decrease in serum HDL cholesterol levels was observed after splenectomy in rats fed normal chow compared with sham-operated rats. An increase in serum triglyceride and a decrease in serum HDL cholesterol levels was observed in both groups of rats fed normal chow plus high-fat cheese. However, these changes were more pronounced in splenectomized rats. These findings suggest that the spleen has a role in lipid metabolism in rats and may therefore influence atherosclerosis.

To identify a possible acute phase response during the steady state of sickle cell disease, we estimated the serum alterations of acute phase proteins, β2‐microglobulin (β2M), κ and λ light chains, interleukins (ILs) and tumor necrosis factor‐α (TNFα) in 21 patients. Increased concentrations of C‐reactive protein (CRP) were found in 5 patients, alpha‐1‐acid‐glycoprotein (AGP) in 3, alpha‐1‐antitrypsin (AAT) in 8, ceruloplasmin (CER) in 2, alpha‐2‐macrogiobulin (AMG) in 14 and decreased haptoglobin (HPT) and transferrin (TFR) in 11 and 9, respectively. Increased β2M was found in 10 patients and κ and λ light chains in 11. IL‐1β, IL‐2, IL‐4, IL‐10 and TNFα were not detected in any of the patients. However, significantly increased values of IL‐6 and sIL‐2r were found. This study has demonstrated increased serum levels of some of the acute phase proteins in patients during the steady state of sickle cell disease. This may be a result of a subclinical vaso‐occlusion which in turn leads to a covert inflammatory response. Cytokines, and in particular IL‐6, produced after this response, seem to be responsible for the high levels of acute phase proteins in the steady state of this disease.