Bradykinin stimulates protein kinase D-mediated colonic myofibroblast migration via cyclooxygenase-2 and heat shock protein 27.

BACKGROUND: Inflammatory bowel disease is characterized by episodic intestinal injury and repair. Myofibroblasts are gastrointestinal tract stromal cells that regulate the reparative process and are known targets of inflammatory mediators including bradykinin (BK). However, the mechanisms through which inflammation regulates myofibroblast-induced wound healing remain incompletely understood. Here, we demonstrate, for the first time, that BK stimulates myofibroblast migration through protein kinase D (PKD)-mediated activation of the cyclooxygenase-2 (COX-2) and heat shock protein 27 (Hsp27) pathways. MATERIALS AND METHODS: CCD-18Co is a human colonic myofibroblast cell line used from passages 8 to 14. An in vitro scratch assay assessed the effect of BK (100 nM) on myofibroblast migration over 24 h in the presence or absence of several inhibitors (CID755673 [10 µM] and NS398 [10 µM]). Hsp27 small interfering RNA evaluated the effect of Hsp27 on colonic myofibroblast migration. Antibodies to pPKD, pHsp27, and COX-2 evaluated expression levels by Western blot. RESULTS: BK stimulated myofibroblast migration over 24 h. BK also led to rapid and sustained phosphorylation of PKD at Ser-916, rapid phosphorylation of Hsp27 at Ser-82, and increased COX-2 expression over 4 h. BK-mediated COX-2 expression and Hsp27 phosphorylation were both inhibited by the PKD inhibitor CID755673. Similarly, BK-induced myofibroblast migration was significantly inhibited by CID755673 (P < 0.05), by the direct COX-2 inhibitor NS398 (P < 0.05), and by Hsp27 small interfering RNA (P < 0.05). CONCLUSIONS: BK stimulates myofibroblast migration through PKD-mediated activation of COX-2 and Hsp27. PKD, COX-2, and Hsp27 all appear to regulate myofibroblast cell migration, a stromal population that may play an important role in mucosal healing in the setting of inflammation.

TNF-α stimulates colonic myofibroblast migration via COX-2 and Hsp27.

BACKROUND: Crohn's disease (CD) is a chronic inflammatory enteropathy characterized by fibrotic strictures. Myofibroblasts (MFBs) are stromal cells of the gastrointestinal tract found in increased numbers in patients with CD and represent the key effector cells involved in pathologic fibrosis. MFB is a known target of tumor necrosis factor alpha (TNF-α), a proinflammatory cytokine strongly implicated in the pathophysiology of CD. However, the precise mechanisms through which TNF-α contributes to fibrosis remain incompletely understood. Here, we demonstrate for the first time that TNF-α increases MFB migration through the cyclooxygenase 2 (COX-2) and heat-shock protein 27 (Hsp27) pathways. MATERIALS AND METHODS: The human colonic MFB cell line 18Co was grown to confluence on 35 × 10 mm cell culture dishes and used from passages 8-14. An in vitro scratch assay assessed the effect of TNF-α (10 ng/mL) on MFB migration over 24 h in the presence or absence of several inhibitors (NS398, SB203580, Hsp27 siRNA). RESULTS: TNF-α significantly increased MFB migration over 24 h. TNF-α also led to the increased expression of COX-2 and stimulated rapid phosphorylation of Hsp27 at serine 82. TNF-α-induced COX-2 expression, Hsp27 phosphorylation, and MFB migration were all significantly inhibited by the P38 MAPK inhibitor SB203580 (P < 0.05). TNF-α-induced MFB migration was also significantly inhibited by NS398 (P < 0.05), a direct inhibitor of COX-2, and by siRNA targeting Hsp27 (P < 0.05). CONCLUSIONS: TNF-α stimulates colonic MFB migration through P38 MAPK-mediated activation of COX-2 and Hsp27. Further elucidating these inflammatory signaling pathways may lead to novel therapeutic targets for the treatment of CD-related fibrosis and strictures.

Regional-specific effects of ovarian hormone loss on synaptic plasticity in adult human APOE targeted replacement mice.

The human apolipoprotein ε4 allele (APOE4) has been implicated as one of the strongest genetic risk factors associated with Alzheimer's disease (AD) and in influencing normal cognitive functioning. Previous studies have demonstrated that mice expressing human apoE4 display deficits in behavioral and neurophysiological outcomes compared to those with apoE3. Ovarian hormones have also been shown to be important in modulating synaptic processes underlying cognitive function, yet little is known about how their effects are influenced by apoE. In the current study, female adult human APOE targeted replacement (TR) mice were utilized to examine the effects of human APOE genotype and long-term ovarian hormone loss on synaptic plasticity in limbic regions by measuring dendritic spine density and electrophysiological function. No significant genotype differences were observed on any outcomes within intact mice. However, there was a significant main effect of genotype on total spine density in apical dendrites in the hippocampus, with post-hoc t-tests revealing a significant reduction in spine density in apoE3 ovariectomized (OVX) mice compared to sham operated mice. There was also a significant main effect of OVX on the magnitude of LTP, with post-hoc t-tests revealing a decrease in apoE3 OVX mice relative to sham. In contrast, apoE4 OVX mice showed increased synaptic activity relative to sham. In the lateral amygdala, there was a significant increase in total spine density in apoE4 OVX mice relative to sham. This increase in spine density was consistent with a significant increase in spontaneous excitatory activity in apoE4 OVX mice. These findings suggest that ovarian hormones differentially modulate synaptic integrity in an apoE-dependent manner within brain regions that are susceptible to neurophysiological dysfunction associated with AD.

Trek to MDG 4: state of Indian States.

OBJECTIVES: To provide projections of progress towards the national and state specific MDG 4 using infant mortality rates (IMR) as the indicator. METHODS: Infant mortality rates (IMR) of major Indian states for year 1990 were used as the base for evaluating their progress in child health. In the absence of any specific guidelines, the state specific target IMR was derived from the IMR:U5MR (under 5 mortality rate) of the countries whose current U5MR is between 11 and 47 per 1,000 live-births (range of target U5MR for Indian states). The projected IMR for year 2015 was then estimated by the average annual rate of reduction (AARR) from 2005 to 2012. RESULTS: Only a few major states-Karnataka, Maharashtra, Odisha, Punjab, and Tamil Nadu are likely to achieve their respective target IMR within the stipulated time (2015). The other major states, and India as a whole, are likely to miss the MDG 4. The two worst performers, Assam and West Bengal, are likely to achieve their respective targets by 2032 and 2022 respectively. Almost all the states have witnessed a significant progress since the advent of National Rural Health Mission (NRHM) in mid-2005-the AARR has almost doubled in the post-NRHM epoch for most states and India as a whole. CONCLUSIONS: The overall progress of most Indian states towards achieving MDG 4 is presently unsatisfactory. However, given the momentum gained since the commencement of NRHM, acceleration in child survival is quite possible in these states.

Discrepancies in reporting the vertebral level of abnormality in MR of the spine.

PURPOSE: The aim of this study was to assess discrepancies in the spinal levels of abnormalities stated in the findings or impression (or both) sections of radiology reports of magnetic resonance (MR) imaging. MATERIALS AND METHODS: Radiology reports from January 2006 through December 2007 (n = 2,097,966) were analyzed using an online radiology report search engine. Reports were searched for presence of the key words MR spine and addendum. The addended reports were then manually assessed for any discrepancies in the reported spinal levels between the body and impression sections; the addenda corrected these errors (identified errors). In addition, all reports with the search term MR spine from January 2006 (n = 1,183) and January 2007 (n = 1,354) were assessed manually to recognize unidentified errors in spinal locations of reported pathology. Two neuroradiologists independently graded the clinical significance of errors on a 5-point scale (1 = definitely not significant, 5 = definitely significant). RESULTS: Of the 11,427 spinal MR reports analyzed in 2006, 7 had identified errors in the sites (levels of the spine) of the lesions. In 2007 (n = 11,785 spinal MR reports), 4 reports were detected with identified errors in spinal levels. In January 2006 and January 2007, 8 and 12 reports, respectively, had unidentified erroneous vertebral levels. Errors were related to discrepant vertebral regions (eg, cervical vs thoracic) in 16% of cases (5 of 31), the wrong number of vertebrae (eg, L2 instead of L3) in 68% of cases (21 of 31), and both in 16% of cases (5 of 31). The average time taken to issue an addendum was 5 +/- 7 days in 2006 and 11 +/- 13 days in 2007. Fifteen reports (48%) scored <3 on the scale of clinical significance, 1 report scored 3, and 15 scored >3. CONCLUSIONS: Errors in lesion level on spinal MR do occur in radiology reports. The number of unidentified errors is substantially higher than that of identified errors. Care should be taken before signing off on radiology reports to identify erroneous mentions of the vertebral levels of abnormalities.

Errare humanum est: frequency of laterality errors in radiology reports.

OBJECTIVE: The purpose of our study was to determine the frequency of discrepancies in side of findings (side discrepancies) in the body and the impression sections of radiology reports. In addition, we determined the frequency of corrected side discrepancies between radiology reports and images in a larger database of radiology reports. MATERIALS AND METHODS: In this cross-sectional study, all radiology reports from January 1, 2007, through December 31, 2007 (n = 1,065,322), that had an addendum were assessed using a radiology reports search engine and searching for the words "left," "right," and "addendum" to locate laterality errors between the body of the report and the impression section. For reports with discrepancies, we recorded the patient's sex, true side of the lesion, and the imaging technique. All reports with an addendum (n = 13,821) in January 2007 containing the words "left" and "right" were evaluated using the same search engine for similar discrepancies. Imaging studies of reports with errors were reviewed to determine the correct side of the lesion and the clinical significance of the errors. RESULTS: Of the 1,065,322 reports, 88 side discrepancies were reported in addenda. The errors in mislabeling the side of the lesion were more common in female (n = 58) than in male (n = 30) patients. Of the 88 errors reported in addenda, 27 were labeled incorrectly in the body of the report, 29 in the impression section, and 32 in both the body and impression sections. In January 2007, 36 of 13,821 (0.26%) reports had no correction for mislabeling of side. Most errors (70.9%) were graded as clinically important. CONCLUSION: Clinically significant errors were found in reporting the body side of the lesion in radiology reports between the body and impression sections.