Angiotensin II induces Fat1 expression/activation and vascular smooth muscle cell migration via Nox1-dependent reactive oxygen species generation.

Fat1 is an atypical cadherin that controls vascular smooth muscle cell (VSMC) proliferation and migration. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 1 (Nox1) is an important source of reactive oxygen species (ROS) in VSMCs. Angiotensin II (Ang II) induces the expression and/or activation of both Fat1 and Nox1 proteins. This study tested the hypothesis that Ang II-induced Fat1 activation and VSMC migration are mediated by Nox1-dependent ROS generation and redox signaling. Studies were performed in cultured VSMCs from Sprague­Dawley rats. Cells were treated with Ang II (1 µmol/L) for short (5 to 30 min) or long term stimulations (3 to 12 h) in the absence or presence of the antioxidant apocynin (10 µmol/L), extracellular-signal-regulated kinases 1/2 (Erk1/2) inhibitor PD98059 (1 µmol/L), or Ang II type 1 receptor (AT1R) valsartan (1 µmol/L). siRNA was used to knockdown Nox1 or Fat1. Cell migration was determined by Boyden chamber assay. Ang II increased Fat1 mRNA and protein levels and promoted Fat1 translocation to the cell membrane, responses that were inhibited by AT1R antagonist and antioxidant treatment. Downregulation of Nox1 inhibited the effects of Ang II on Fat1 protein expression. Nox1 protein induction, ROS generation, and p44/p42 MAPK phosphorylation in response to Ang II were prevented by valsartan and apocynin, and Nox1 siRNA inhibited Ang II-induced ROS generation. Knockdown of Fat1 did not affect Ang II-mediated increases in Nox1 expression or ROS. Inhibition of p44/p42 MAPK phosphorylation by PD98059 abrogated the Ang II-induced increase in Fat1 expression and membrane translocation. Knockdown of Fat1 inhibited Ang II-induced VSMC migration, which was also prevented by valsartan, apocynin, PD98059, and Nox1 siRNA. Our findings indicate that Ang II regulates Fat1 expression and activity and induces Fat1-dependent VSMC migration via activation of AT1R, ERK1/2, and Nox1-derived ROS, suggesting a role for Fat1 downstream of Ang II signaling that leads to vascular remodeling.

Use of Sysmex XE-2100 and XE-5000 hematology analyzers for the diagnosis of malaria in a nonendemic country (France).

INTRODUCTION: Most studies dealing with automated hematology analyzers (HAs) and malaria diagnosis are conducted in endemic countries. METHODS: We retrospectively studied cell blood counts (CBCs) performed with Sysmex XE-2100 and XE-5000 HAs in our center (Angers, France) regarding 67 patients returning from endemic areas and infected with various Plasmodium species. RESULTS: In 83% of infected samples with Plasmodium vivax (Pv), ovale (Po), or malariae (Pm), extra clouds of dots were present in neutrophil and/or eosinophil area(s) on routine differential (DIFF) scattergrams. In contrast, samples infected with Plasmodium falciparum (Pf) failed to show such DIFF scattergrams, or any other suggesting malaria infection (0/ 49 pts). Abnormal areas from DIFF scattergrams were related to the presence of mature schizonts and gametocytes, undestroyed by lysis agent, the latter not observed in Pf-infected patients from our series. The internal parameter WBC[DIFF] - WBC[BASO] raised in parallel to parasitemia in Pv, Po, and Pm samples but could not be used as a surrogate for parasitemia. In Pf infection, reticulocyte/ immature reticulocyte fraction (IRF) ratio showed a significant correlation with parasitemia (P < 0.05). A diagnostic model developed for Pf in endemic countries showed sensitivity of 77%. CONCLUSION: Using SYSMEX analyzers, Pv, Po, and Pm infections are easy to ascertain as DIFF scattergrams are almost specific (specificity = 99.9%). Pf infection diagnosis by CBC may be a more promising tool.

Gram-negative bacteremia: which empirical antibiotic therapy?

PURPOSE: Given the increasing frequency of cefotaxime-resistant strains, third-generation cephalosporins (3GC e.g. cefotaxime, ceftriaxone) might not be recommended any longer as empirical antibiotic therapy for community-acquired Gram-negative bacteremia (CA-GNB). PATIENTS AND METHODS: We conducted a multicenter prospective descriptive study including patients with CA-GNB. RESULTS: Two hundred and nineteen patients were included. Escherichia coli and Pseudomonas aeruginosa were the most frequently isolated species in 63% (n=138) and 11% (n=24) of the cases, respectively. The prevalence of cefotaxime-resistance reached 18% (n=39) mostly due to intrinsic resistance (27 cases, 12%). The presence of invasive material (P<0.001), the origin of the patient (Paris region or West of France) (P=0.006), and home health care (P<0.001) were variables predicting resistant GNB. The negative predictive value for resistance in patients with invasive material coming from the West of France, or without invasive material and with home health care was 94%. The positive predictive value for patients with invasive material living in Paris, or without invasive material and with home health care only reached 58 and 54%, respectively. CONCLUSIONS: Using 3GC for CA-GNB due to cefotaxime-resistant strains was relatively frequent, ESBL-producing Enterobacteriaceae being rarely involved. Our study highlights the role of local epidemiology; before any changes to first-line antibiotic therapy, local epidemiological data should be taken into account.

Pyrrolidine dithiocarbamate down-regulates vascular matrix metalloproteinases and ameliorates vascular dysfunction and remodelling in renovascular hypertension.

BACKGROUND AND PURPOSE: Mounting evidence implicates matrix metalloproteinase (MMP) in the vascular dysfunction and remodelling associated with hypertension. We tested the hypothesis that treatment with pyrrolidine dithiocarbamate (PDTC), which interferes with NF-κB-induced MMPs gene transcription, could exert antihypertensive effects, prevent MMP-2 and MMP-9 up-regulation, and protect against the functional alterations and vascular remodelling of two-kidney, one clip (2K1C) hypertension. EXPERIMENTAL APPROACH: Sham-operated or hypertensive rats were treated with vehicle or PDTC (100 mg·Kg(-1) ·day(-1)) by gavage for 8 weeks. Systolic blood pressure (SBP) was monitored weekly. Aortic rings were isolated to assess endothelium-dependent relaxations. Quantitative morphometry of structural alterations of the aortic wall was carried out in haematoxylin/eosin sections. Formation of vascular reactive oxygen species (ROS), and inducible (i) NOS and phosphorylated-p65 NF-κB subunit expression were measured in the aortas. MMP-2 and MMP-9 aortic levels and gelatinolytic activity were determined by gelatin and in situ zymography and by immunofluorescence. KEY RESULTS: Treatment with PDTC attenuated the increases in SBP and prevented the endothelial dysfunction associated with 2K1C hypertension. Moreover, PDTC reversed the vascular aortic remodelling, the increases in aortic ROS levels and in iNOS and phosphorylated-p65 NF-κB expression found in 2K1C rats. These effects were associated with attenuation of 2K1C up-regulation of aortic MMP-2 and MMP-9 levels and gelatinolytic activity. CONCLUSION AND IMPLICATIONS: These findings suggest that PDTC down-regulates vascular MMPs and ameliorates vascular dysfunction and remodelling in renovascular hypertension, thus providing evidence supporting the suggestion that PDTC is probably a good candidate to be used to treat hypertension.

Development of thyroid disease in patients with primary and secondary Sjögren's syndrome.

Autoimmune thyroid disease associated to Sjögren syndrome (SS) may be defined by serological and functional abnormalities. We studied the prevalence of serum anti-thyroid autoantibodies and the development of thyroid functional defects by a follow-up investigation in patients with primary (pSS) and secondary SS (sSS). In keeping with previous literature data, our findings confirmed an increased prevalence of thyroid autoimmune phenomena in the whole series of patients with SS. In addition, evidence was provided for a greater incidence of serological and functional thyroid abnormalities in pSS when compared to sSS. Follow-up study also showed that the rate of thyroid autoimmune disease development was higher in pSS than in sSS. The appearance of circulating anti-thyroid antibodies was particularly frequent in patients with preexisting antibodies against extractable nuclear antigens (anti-ENA). These data indicate the need for a careful monitoring of thyroid function in SS patients, with particular regard to those with pSS and positive anti-ENA tests.