Decreased expression of profilin 2 in oral squamous cell carcinoma and its clinicopathological implications.

Profilins are small proteins essential for many normal cellular dynamics and constitute one of the crucial components of actin-based cellular motility. Several recent studies have implicated a role for the profilin (PFN) family in cancer pathogenesis and progression. However, their expression and promising functions are largely unknown in oral squamous cell carcinoma (OSCC). In this study, we analyzed the correlation between PFN1 and PFN2 expression in vitro and in vivo. The protein expression levels were roughly compared between cell lines (HIOEC, HB96) with the employment of mass spectrometry. PFN2 was singled out as one of the significantly down-regulated genes in the cancerous HB96 cells. The expression levels of PFN1 and PFN2 in vitro were validated by RT-PCR, real-time PCR and Western blotting. Laser scanning confocal microscopy was used for the first time to assess the localization of PFN2 expression. In subsequent experiments, we observed the relationship between PFN2 expression levels and the proliferation of transfected HB96 cancer cells. VASP, N-WASP and P27 expression was also examined in the PFN2-transfected or non-transfected HB96 cells. In vivo, antigen expression was determined by immunohistochemical analyses in 88 paired tissue specimens. Decreased protein expression was confirmed in cancerous tissues from 88 OSCC patients compared with paracancerous normal mucous epithelia. Tumors with weak PFN2 expression were associated with a significantly worse prognosis than strongly expressed tumours (P<0.001). Other statistical analyses were performed to assess the differences in expression and their clinical and pathological significance. In conclusion, PFN2 can be utilized as both a potential suppressor marker and a prognostic protein for OSCC. The function of PFN2 may be to regulate the N-WASP/Arp2/3 signaling pathway.

Incidence of sterile cerebrospinal fluid pleocytosis in infants with urinary tract infection.

AIM: To determine the incidence of sterile cerebrospinal fluid (CSF) pleocytosis in infants ≤6 months old with urinary tract infection (UTI). METHODS: Retrospective study of children admitted to a tertiary children's hospital in 2006 and 2007 with UTI who also had a lumbar puncture performed. All urine specimens were tested for anti-microbial activity. RESULTS: Twelve (11.3%) of 106 infants with UTI had concurrent CSF pleocytosis. None of these patients had anti-microbial activity in the urine, showing that they had not received prior antibiotics. None of the 15 neonates (≤28 days old) with UTI and lumbar puncture had CSF pleocytosis. Antibiotics were stopped after a maximum of 10 days. CONCLUSION: Our results are compatible with published reports on the proportion of infants with UTI who have concurrent sterile CSF pleocytosis. We were able to exclude previous antibiotic therapy by measuring urinary anti-microbial activity. Our work supports the hypothesis that CSF pleocytosis in UTI is inflammatory and not because of infection of the central nervous system.

Modification of in vivo and in vitro T- and B-cell-mediated immune responses by the Pseudomonas aeruginosa quorum-sensing molecule N-(3-oxododecanoyl)-L-homoserine lactone.

N-3-(oxododecanoyl)-L-homoserine lactone (OdDHL), a quorum-sensing molecule of Pseudomonas aeruginosa, plays an important role in the pathogenesis of the organism through its control of virulence factor expression. Several reports have suggested that OdDHL can also directly modulate host immune responses. However, the nature of the modulation is controversial, with different reports suggesting promotion of either humoral (Th2-mediated) or inflammatory (Th1-mediated) responses. This report describes a series of studies which demonstrate for the first time that in vivo administration of OdDHL can modulate the course of an antibody response, with an increase in ovalbumin (OVA)-specific immunogloblulin G1 (IgG1) but not IgG2a in OdDHL-treated OVA-immunized BALB/c mice compared to levels for controls. In vitro stimulation of lymphocytes from both Th1-biased C57Bl/6 and T-cell receptor transgenic mice and Th2-biased BALB/c mice in the presence of OdDHL demonstrated that OdDHL inhibits in vitro cytokine production in response to both mitogen and antigen, with gamma interferon (IFN-gamma) tending to be more inhibited than interleukin-4 (IL-4). In vitro mitogen or antigen restimulation of cells from mice treated with OdDHL in vivo shows effects on cytokine production which depend on the underlying immune bias of the mouse strain used, with a relative increase of IFN-gamma in Th1-biased C57Bl/6 mice and a relative increase of IL-4 in Th2-biased BALB/c mice. Thus, the mode of action of OdDHL on T-cell cytokine production is likely to be a relatively nonspecific one which accentuates an underlying immune response bias rather than one which specifically targets either Th1 or Th2 responses.