Detection and quantification of infectious avian influenza A (H5N1) virus in environmental water by using real-time reverse transcription-PCR.

Routes of avian influenza virus (AIV) dispersal among aquatic birds involve direct (bird-to-bird) and indirect (waterborne) transmission. The environmental persistence of H5N1 virus in natural water reservoirs can be assessed by isolation of virus in embryonated chicken eggs. Here we describe development and evaluation of a real-time quantitative reverse transcription (RT)-PCR (qRT-PCR) method for detection of H5N1 AIV in environmental water. This method is based on adsorption of virus particles to formalin-fixed erythrocytes, followed by qRT-PCR detection. The numbers of hemagglutinin RNA copies from H5N1 highly pathogenic AIV particles adsorbed to erythrocytes detected correlated highly with the infectious doses of the virus that were determined for three different types of artificially inoculated environmental water over a 17-day incubation period. The advantages of this method include detection and quantification of infectious H5N1 AIVs with a high level of sensitivity, a wide dynamic range, and reproducibility, as well as increased biosecurity. The lowest concentration of H5N1 virus that could be reproducibly detected was 0.91 50% egg infective dose per ml. In addition, a virus with high virion stability (Tobacco mosaic virus) was used as an internal control to accurately monitor the efficiency of RNA purification, cDNA synthesis, and PCR amplification for each individual sample. This detection system could be useful for rapid high-throughput monitoring for the presence of H5N1 AIVs in environmental water and in studies designed to explore the viability and epidemiology of these viruses in different waterfowl ecosystems. The proposed method may also be adapted for detection of other AIVs and for assessment of their prevalence and distribution in environmental reservoirs.

Evaluation of two surgical techniques for acetabular reconstruction in total hip replacement for congenital hip disease: results after a minimum ten-year follow-up.

We have evaluated the results of total hip replacement in patients with congenital hip disease using 46 cemented all-polyethylene Charnley acetabular components implanted with the cotyloplasty technique in 34 patients (group A), and compared them with 47 metal-backed cementless acetabular components implanted without bone grafting in 33 patients (group B). Patients in group A were treated between 1988 and 1993 and those in group B between 1990 and 1995. The mean follow-up for group A was 16.6 years (12 to 18) and the mean follow-up for group B was 13.4 years (10 to 16). Revision for aseptic loosening was undertaken in 15 hips (32.6%) in group A and in four hips (8.5%) in group B. When liner exchange was included, a total of 13 hips were revised in group B (27.7%). The mean polyethylene wear was 0.11 mm/yr (0.002 to 0.43) and 0.107 mm/yr (0 to 0.62) for groups A and B, respectively. Polyethylene wear in group A was associated with linear osteolysis, and in group B with expansile osteolysis. In patients with congenital hip disease, when 80% cover of the implant can be obtained, a cementless acetabular component appears to be acceptable and provides durable fixation. However, because of the type of osteolysis arising with these devices, early exchange of a worn liner is recommended before extensive bone loss makes revision surgery more complicated.

Human ORF (ecthyma contagiosum).

Orf is a viral cutaneous infection typically seen on the hands of people involved in sheep and goat farming. We report 31 cases of orf and consider its pathology, characteristic appearance, diagnosis, treatment and complications.

Survival in a residential home: an eleven-year longitudinal study.

The length of survival and various risk factors were studied utilizing 408 residents (141 men, 267 women) of a large residential home. The subjects, aged 68 years or more at entry, admitted between 1978 and 1983, were physically independent, continent, and non-diabetic. They were followed until December 31, 1988, by which time 78% had died. The multivariate proportional hazard analysis showed the following entry variables to have the indicated effects on relative mortality rate ratios: 5-years' higher age (+10%, NS), persistent bacteriuria (+13%, NS), abnormal ECG (+26%, NS), current smoking (+63%, P less than 0.01), mildly impaired mobility (+96%, P less than 0.001), higher levels of in-study systolic pressure in 10-mm Hg steps (-4%, NS), higher entry serum cholesterol in 1-mmol/L steps (-7%, NS), and higher hematocrit in 5% steps (-14%, P less than 0.02). Female sex was associated with a +25% (NS) rate ratio; socioeconomic status and body weight were without effect. These data highlight the relative importance of specific factors associated with survival of persons within retirement homes and indicate that: (1) the presence of mild impairment of mobility at entry is by far the strongest predictor of early death; (2) smoking and lower hematocrit also exert important adverse effects; and (3) certain "risk factors", ie elevated systolic blood pressure and serum cholesterol, have a minimal protective effect, if any, in this age group.

Cases of swollen head syndrome in broiler chickens in Greece.

From 50 commercial broiler flocks included in a study concerning respiratory disease, signs of swollen head syndrome (SHS) were shown in eight. Postmortem examination was performed in eight birds showing signs of SHS from each flock. The trachea and head from each bird were collected for laboratory investigation. An enzyme-linked immunosorbent assay (ELISA) was used for the detection of viral and avian mycoplasma antigens in the trachea, and bacteriologic examinations were performed from the infraorbital sinuses of the infected birds. According to the ELISA results, the most frequently detected antigen in the trachea was Mycoplasma synoviae (six flocks, 75%), followed by infectious bronchitis virus (IBV) (five flocks, 62.5%), avian adenovirus (four flocks, 50%), avian reovirus (three flocks, 37.5%), Mycoplasma gallisepticum (one flock, 12.5%), and Newcastle disease virus (NDV) (one flock, 12.5%). Turkey rhinotracheitis (TRT), infectious laryngotracheitis, and avian influenza viral antigens were not detected. Experimental assays for characterization of NDV and IBV isolates showed that they were strains of low virulence (evidently vaccine strains). Bacteriologic examinations from the infraorbital sinuses of the affected birds resulted in the isolation of Escherichia coli (seven cases, 87.5%) and Staphylococcus spp. (one case, 12.5%). It is evident that TRT virus did not play a causal role in SHS in commercial broiler flocks in Greece, but in this condition, other viruses (IBV, NDV), mycoplasmas, or bacteria may be involved, and environmental conditions seem to be essential to the occurrence and severity of the disease.

[Antigen HLA-B5 in Adamantiadès-Behçet syndrome (author's transl)]. / Le HLA-B5 antigène dans le syndrome d'Adamantiadès-Behçet, sa valeur diagnostique à propos de 4 cas atypiques).

Among 16 cases clinicaly diagnosed as Adamantiadès-Behçet syndrome coming from various areas of Northern Greece, antigen HLA-B5 was found on 12, i.e. 75% instead of 18,5% in the normal population of the same areas (200 subjects were examined by Dr. Z. Polymenidis in this thesis). On 3 other cases the antigen found was HLA-BW35 (belonging to group C4), while on the last one the antigen was HLA-B27. 4 cases, suspicious for atypical Adamantiadès-Behçet syndrome, are reported in which the discovery of antigen HLA-B5 inhanced the clinical diagnosis of the syndrome and made possible the early application of suitable treatment (steroids, immunosupressors).

Charnley low-friction arthroplasty in patients 35 years of age or younger. Results at a minimum of 23 years.

We evaluated the outcome of 41 consecutive Charnley low-friction arthroplasties (LFAs) performed by a single surgeon in 28 patients aged ≤ 35 years at operation between 23 and 36 years previously. There were 20 women and eight men with a mean age of 32 years (23 to 35) at surgery. Two patients (three hips) were lost to follow-up at 12 and 17 years post-operatively, respectively, and one patient (one hip) died at 13 years post-operatively. These patients were excluded from the final evaluation. The survival rate of the acetabular components was 92.7% (95% confidence interval (CI) 88.7 to 96.7) at ten years, 67.1% (95% CI 59.75 to 74.45) at 20 years and 53.2% (95% CI 45.3 to 61.1) at 25 years. For the femoral component the survival was 95.1% (95% CI 91.8 to 98.5) at ten years, 77.1% (95% CI 73.9 to 80.3) at 20 years and 68.2% (95% CI 60.7 to 75.8) at 25 years. The results indicate that the Charnley LFA remains a reasonable choice in the treatment of young patients and can serve for comparison with newer techniques and implants.

Trochanteric osteotomy in total hip replacement for congenital hip disease.

We studied the effect of trochanteric osteotomy in 192 total hip replacements in 140 patients with congenital hip disease. There was bony union in 158 hips (82%), fibrous union in 29 (15%) and nonunion in five (3%). The rate of union had a statistically significant relationship with the position of reattachment of the trochanter, which depended greatly on the pre-operative diagnosis. The pre-operative Trendelenburg gait substantially improved in all three disease types (dysplasia, low and high dislocation) and all four categories of reattachment position. A persistent Trendelenburg gait post-operatively was noticed mostly in patients with defective union (fibrous or nonunion). Acetabular and femoral loosening had a statistically significant relationship with defective union and the position of reattachment of the trochanter. These results suggest that the complications of trochanteric osteotomy in total hip replacement for patients with congenital hip disease are less important than the benefits of this surgical approach.

An examination of the association between different morphotypes of femoroacetabular impingement in asymptomatic subjects and the development of osteoarthritis of the hip.

We retrospectively examined the long-term outcome of 96 asymptomatic hips in 96 patients with a mean age of 49.3 years (16 to 65) who had radiological evidence of femoroacetabular impingement. When surveillance commenced there were 17, 34, and 45 hips with cam, pincer, and mixed impingement, respectively. Overall, 79 hips (82.3%) remained free of osteoarthritis for a mean of 18.5 years (10 to 40). In contrast, 17 hips (17.7%) developed osteoarthritis at a mean of 12 years (2 to 28). No statistically significant difference was found in the rates of development of osteoarthritis among the three groups (p = 0.43). Regression analysis showed that only the presence of idiopathic osteoarthritis of the contralateral diseased hip was predictive of development of osteoarthritis on the asymptomatic side (p = 0.039). We conclude that a substantial proportion of hips with femoroacetabular impingement may not develop osteoarthritis in the long-term. Accordingly, in the absence of symptoms, prophylactic surgical treatment is not warranted.

Unilateral hip osteoarthritis: can we predict the outcome of the other hip?

OBJECTIVE: The objective of this study was to define, in unilateral hip osteoarthritis (OA), factors predicting the outcome of the other hip. MATERIALS AND METHODS: We examined the anteroposterior radiographs of the pelvis of 95 white patients with unilateral idiopathic (56 patients) or secondary to congenital hip diseases (39 patients) OA. The other hip was free from symptoms (pain or limping) at the initial examination and without radiographic evidence of OA; it was what we call a "normal" hip. Two parameters were evaluated: (1) the type of osteoarthritis of the involved hip and (2) the range of four radiographic indices of the contralateral hip: the sourcil inclination (weight-bearing surface), the acetabular angle, the Wiberg's center-edge angle, and the neck-shaft angle. Follow-up radiographs for the hips that remained OA-free were available for 10 to 35 years and for those that developed OA, at the time of initial symptoms, range 2 to 31 years. RESULTS: Logistic regression analysis showed that the presence of idiopathic OA in one hip had a statistically significant effect on the development of OA on the other hip (p < 0.001). Minor deviations of radiographic indices of the contralateral hip is not a predictive factor for its outcome. When the radiographic indices are examined together with the pathology of the involved hip, only WBS was shown to have a significant effect to the development of OA and its type (p < 0.001). CONCLUSIONS: The following conclusions can be drawn from this study: 1. Patient with idiopathic OA of one hip is at increased risk of developing OA in the other hip. 2. The outcome of the other hip cannot be predicted only on the basis of the evaluation of its radiographic indices. 3. Among the different indices, WBS seems to have a strong influence toward the development of OA.

Mid-term behaviour of cemented titanium stems: A seven to eleven year clinical.

A controversy exists regarding the results of cemented Ti-alloy femoral stems. Although most authors reported unfavourable results, there is no clear evidence of the exact reason for the increased failure rate. We studied 35 arthroplasties in 30 patients with a mean age at the time of surgery of 49.7 years (range 27 to 67 years). In most of them (23 hips) the pre-operative diagnosis was hip dysplasia or dislocation. The femoral component used in this study was the Optifix stem. Survival analysis showed a cumulative rate of success at nine years of 90.4% (15.3) with 25.5 femoral components remaining at risk. Three femoral components were retrieved during revision surgery: due to fatigue fracture, fracture of the femur and aseptic loosening respectively. Given that most publications refer to different designs of stems we presume that the problem could be focused on the design rather than the material or the method of fixation. Further information is necessary before Ti-alloy stems fixed with cement are withdrawn from use. (Hip International 2005; 15: 12-20).

Temperature and serum proinflammatory cytokine changes in patients with NSCLC after BAL.

We examined the effects of bronchoalveolar lavage (BAL) and BAL fluid characteristics on the systemic proinflammatory cytokine expression and their relation to clinical and laboratory findings. Thirty patients suspected to have lung cancer were subjected to fiber-optic bronchoscopy (FOB) and BAL. Clinical and laboratory findings were determined at baseline, 4 h, and 24 h, including lung auscultation, temperature, chest X-ray, WBC, neutrophils, and serum IL-1beta, IL-6, and TNF-alpha. BAL fluid characteristics were determined including cytokine levels. Fifteen volunteers served as controls to determine serum variation of the same cytokines. Significant temperature elevation was defined as 1 degrees C increase compared to baseline. BAL was associated with temperature and serum TNF-alpha and IL-6 but not IL-1beta increase at 4 h. Four patients (13.3%) developed temperature over 38 degrees C. In controls there were no significant changes between baseline and 24 h measurements for the same cytokines. Eleven patients (36.6%) developed a significant temperature elevation 4 h after BAL. These patients had a statistically significant ( p < 0.05) increase in serum IL-6 at 4 h and in TNF-alpha at both 4 and 24 h after BAL compared with the nonsignificant temperature increase group. BAL characteristics were not different between the two groups. On the other hand, BAL fluid IL-6 and TNF-alpha levels were significantly higher ( p < 0.05) in the nonfever group. Significant temperature increase was observed in 36.6% of the patients undergoing BAL and associated with significant serum TNF-alpha and IL-6 increase at 4 h. Lung cytokines levels, alveolar macrophages, and BAL fluid characteristics are not related to temperature and serum proinflammatory cytokine increase. The hypothesis of alveolar macrophages derive from cytokine production and shift to the systemic circulation cannot be supported by our data.

Surgical treatment of gastric impaction in farmed ostriches.

Partial or complete impaction of the proventriculus and/or the ventriculus is usually seen in juvenile ostriches. Common impaction materials include stones, sand, hay stems and leaves, and even plastic and metallic objects. Six live ostriches showing signs of impaction and three dead ones suspected to have impacted stomachs were presented to the Department of Clinical Sciences of the Faculty of Veterinary Medicine of the Aristotle University of Thessaloniki. The birds manifested anorexia, whitish and decreased urine output, scant defecation, unwillingness to rise and consume water. Clinical examination revealed distended and firm proventriculi, while radiographs showed foreign objects in the proventriculi of all birds and the ventriculus of one of them. The living birds were successfully treated surgically and medically. Autopsy of the three dead ostriches revealed severe dehydration and impaction of the stomachs. Bacteriological and parasitological examination was negative. Amendments in management practices were important to minimize losses caused by impaction of the stomachs in ostriches, while proventriculotomy remains the most effective approach to the problem.

Biological active peptides in human urine: III. Inhibitors of the growth of human leukemia, osteosarcoma, and HeLa cells.

Medium-sized peptides isolated from normal humans urine were tested for their effect on DNA, RNA, and protein synthesis, and mitosis, in tissue culture of human myeloblastic leukemia, osteosarcoma, and HeLa cells. Two types of antineoplastic peptides were found. One type consists of strongly acidic peptides (probably sulfated glycopeptides) which act specifically on different kinds of neoplasma. The other type comprises slightly acidic and neutral peptides, and has broad specificity. The active peptides produce up to 97% inhibition of DNA synthesis and mitosis in the neoplastic cells in tissue culture. The peptide fraction which has broad specificity was tested in different concentrations and gave good dose-response relationship.