Search

Background: Streptococcus pneumoniae and Haemophilus influenzae cause pneumonia and as Neisseria meningitidis they are important agents of meningitis. Although several PCR methods have been described for these bacteria the specificity is an underestimated problem. Here we present a quantitative multiplex real-time PCR (qmPCR) for detection of S. pneumoniae (9802 gene fragment), H. influenzae (omp P6 gene) and N. meningitidis (ctrA gene). The method was evaluated on bronchoalveolar lavage (BAL) samples from 156 adults with lower respiratory tract infection (LRTI) and 31 controls, and on 87 cerebrospinal fluid (CSF) samples from meningitis patients. Results: The analytical sensitivity was not affected by using a combined mixture of reagents and a combined DNA standard (S. pneumoniae/H. influenzae/N. meningitidis) in single tubes. By blood- and BAL-culture and S. pneumoniae urinary antigen test, S. pneumoniae and H. influenzae were aetiological agents in 21 and 31 of the LTRI patients, respectively. These pathogens were identified by qmPCR in 52 and 72 of the cases, respectively, yielding sensitivities and specificities of 95% and 75% for S. pneumoniae, and 90% and 65% for H. influenzae, respectively. When using a cut-off of 105 genome copies/mL for clinical positivity the sensitivities and specificities were 90% and 80% for S. pneumoniae, and 81% and 85% for H. influenzae, respectively. Of 44 culture negative but qmPCR positive for H. influenzae, 41 were confirmed by fucK PCR as H. influenzae. Of the 103 patients who had taken antibiotics prior to sampling, S. pneumoniae and H. influenzae were identified by culture in 6% and 20% of the cases, respectively, and by the qmPCR in 36% and 53% of the cases, respectively. In 87 CSF samples S. pneumoniae and N. meningitidis were identified by culture and/or 16 S rRNA in 14 and 10 samples and by qmPCR in 14 and 10 samples, respectively, giving a sensitivity of 100% and a specificity of 100% for both bacteria. Conclusions: The PCR provides increased sensitivity and the multiplex format facilitates diagnosis of S. pneumoniae, H. influenzae and N. meningitidis and the assay enable detection after antibiotic treatment has been installed. Quantification increases the specificity of the etiology for pneumonia. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Interleukin-1beta (IL-1beta) is a key cytokine linked to the pathogenesis of acute arthritis. Caspase 1, neutrophil elastase, and chymase all process proIL-1beta to its biologically active form. This study was undertaken to examine the potential contributions of each of these proteases in experimental models of inflammatory arthritis. METHODS: Caspase 1-deficient (Casp1(-/-)) and wild-type (WT) mice were tested for their response to arthritogenic K/BxN serum transfer for induction of arthritis or injection of monosodium urate monohydrate (MSU) crystals for induction of peritonitis. All mice were prophylactically treated with inhibitors of neutrophil elastase or chymase. Arthritic paws were tested for the presence of IL-1beta protein by enzyme-linked immunosorbent assay and Western blotting. Neutrophils and mast cells from WT and mutant mice were tested for their ability to secrete IL-1beta after in vitro stimulation, in the presence of protease inhibitors. RESULTS: Casp1(-/-) and WT mice developed paw swelling to the same extent in the K/BxN serum transfer-induced arthritis model. MSU crystal injection into Casp1(-/-) mice also resulted in neutrophil influx and production of measurable peritoneal IL-1beta protein. Both of these responses were attenuated with neutrophil elastase inhibitors. K/BxN serum transfer-induced arthritis was also reduced by treatment with a chymase inhibitor. Casp1(-/-) neutrophils and mast cells, when exposed to MSU crystals, secreted similar amounts of IL-1beta protein upon in vitro stimulation with lipopolysaccharide, albeit at lower levels than that secreted by WT cells. Elastase and chymase inhibitors reduced the amount of IL-1beta released by these cells. CONCLUSION: The production of IL-1beta by neutrophils and mast cells is not exclusively dependent on caspase 1, and other proteases can compensate for the loss of caspase 1 in vivo. These pathways might therefore compromise the caspase 1-targeted therapies in neutrophil-predominant arthritis. [ABSTRACT FROM AUTHOR]

In healthy blood donors, serological positivity for human cytomegalovirus (HCMV) is associated with an increased proportion of NK cells bearing the CD94/NKG2C NK cell receptor (NKR). The expression of the activating CD94/NKG2C NKR and of the inhibitory CD94/NKG2A NKR was studied in a cohort of 45 aviremic human immunodeficiency virus type 1 (HIV-1)-positive patients receiving highly active antiretroviral therapy. The proportions of NKG2C(+) NK cells were significantly increased in HIV-1-positive patients (mean+/-SD, 25.9%+/-23.0%), compared with those in 31 healthy individuals (mean+/-SD, 16.1%+/-20.7%). Yet, the association vanished when HCMV serological status was considered in a multivariate regression model. These results support the conclusion that changes in the NKR repertoire in HIV1-positive patients are related to a concomitant HCMV infection. Copyright © 2006 Infectious Diseases Society of America [ABSTRACT FROM AUTHOR]

Introduction/Background The Osteoarthritis Research Society International recommends evaluating physical function in gonarthrosic patients after a total knee arthroplasty using tests such as timed up and go (TUG) and six-minute walking test (6MWT). In 1991, Berman et al. found that strength was not fully recovered until two years after surgery. Our aim was to demonstrate the results in physical function and isokinetic muscular strength of gonarthrosis patients prior to undergoing surgery and three months after a total knee arthroplasty (TKA). Material and method Prospective study (September 2016–December 2017) of patients with gonarthrosis awaiting a total knee arthroplasty surgery. Variables recorded: gender, age, dominant hand, operated side, body mass index (BMI), walking aids. Measured outcomes before surgery and three months afterwards: Visual Analogue Scale (VAS) at rest and active, TUG, 6MWT and mean peak torque (PT) of isokinetic strength of quadriceps and hamstrings between the TKA side and the unaffected knee at 60°/s. Data analysis of means and percentages. Means compared using Student t and non-parametric tests. Statistical significance was set at P < 0.05. Results Forty-four patients, 36.4% male. Mean age 67.8 ± 6.5. Mean BMI 30.3 ± 6.7; 63.6% of the individuals used walking aids before surgery and 84.1% did not use them after surgery. The results obtained on VAS and physical function prior to surgery and three months after the intervention were. A decrease of strength in the affected quadriceps and a significant decrease of strength in the affected hamstrings were observed. However the strength of the unaffected quadriceps increased slightly and the unaffected hamstrings strength did not vary. Conclusion Three months after a TKA surgery, even though there was a descent in muscular strength in the affected limb, our patients experienced an improvement in physical function possibly due to pain decrease. [ABSTRACT FROM AUTHOR]