In inflammatory bowel disease, dysregulated T cells express pro-inflammatory cytokines. Using a chronic azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced colitis model resembling ulcerative ...colitis, we evaluated whether and when treatment with the Janus kinase (JAK) inhibitor tofacitinib could be curative. Comparing the treatment with two and three cycles of tofacitinib medication in drinking water - intermittently with DSS induction - revealed that two cycles were not only sufficient but also superior over the 3-x regimen. The two cycles of the 2-x protocol paralleled the second and third cycles of the longer protocol. T cells were less able to express interferon gamma (IFN-γ) and the serum levels of IFN-γ, interleukin (IL)-2, IL-6, IL-17, and tumor necrosis factor (TNF) were significantly reduced in sera, while those of IL-10 and IL-22 increased under the 2-x protocol. Likewise, the frequency and effector phenotype of regulatory T cells (Tregs) increased. This was accompanied by normal weight gain, controlled clinical scores, and restored stool consistency. The general and histologic appearance of the colons revealed healing and tissue intactness. Importantly, two phases of tofacitinib medication completely prevented AOM-incited pseudopolyps and the hyper-proliferation of epithelia, which was in contrast to the 3-x regimen. This implies that the initial IBD-induced cytokine expression is not necessarily harmful as long as inflammatory signaling can later be suppressed and that time-restricted treatment allows for anti-inflammatory and tissue-healing cytokine activities.
Aims
To compare in vivo DNA lesions induced during helical and sequential coronary computed tomography angiography (CTA) and to evaluate the effect of CT parameters on double-strand break (DSB) ...levels.
Methods
Thirty-six patients were examined with various CT protocols and modes (helical scan,
n
= 27; sequential scan,
n
= 9) either using a 64-slice dual-source or a 128-slice CT system. Blood samples were obtained before and 30 min after CT. Lymphocytes were isolated, stained against the phosphorylated histone variant γ-H2AX, and DSBs were visualised by using fluorescence microscopy.
Results
DSB yields 30 min after CTA ranged from 0.04 to 0.71 per cell and showed a significant correlation to DLP (
ρ
= 0.81,
p
< 0.00001). Median DSB yield and median DLP were significantly lower after sequential compared to helical CT examinations (0.11 vs. 0.37 DSBs/cell and 249 vs. 958 mGy cm,
p
< 0.00001). Additional calcium scoring led to an increase in DLP (
p
= 0.15) and DSB levels (
p
= 0.04). DSB levels normalised to the DLP showed a significant correlation to the attenuation of the blood (
ρ
= 0.53,
p
= 0.01) and a negative correlation to the body mass index of the patients (
ρ
= −0.37,
p
= 0.06).
Conclusion
γ-H2AX immunofluorescence microscopy allows one to determine dose-related effects on x-ray-induced DSB levels and to consider individual factors which cannot be monitored by physical dose measurements.
Background and Aims
Nitrospirillum amazonense
strain CBAmC is a nitrogen-fixing bacterium that has been used for sugarcane inoculation. Despite its beneficial effects, however, little is known about ...the establishment and colonization of this microorganism in its inoculated host plant. Furthermore, alternative inoculation methods need to be tested in the sugarcane crop. The objective of the present work was to quantify and visualize CBAmC in sugarcane cultivar RB867515 after using three inoculation methods.
Methods
Utilizing one node as propagation material, three bacterial inoculation methods were tested under greenhouse conditions. For up to 60 days after inoculation (DAI), qPCR was used to quantify the bacterial loads in the plant or substrate while FISH-CLSM was used to confirm the establishment and colonization profile of CBAmC in different plant tissues.
Results
FISH-CLSM indicated that the CBAmC strain could establish itself on different tissues of sugarcane, especially roots. qPCR revealed variable bacterial loads in the plants inoculated according to the three methods tested. In addition, little or no bacterial loads were detected in the substrate a few days after inoculation, regardless of the method used.
Conclusions
N. amazonense
CBAmC proves to be a bacterium that preferentially associates with the surface of the root tissues of sugarcane propagated by one node, instead of colonizing internal plant tissues or remaining in the soil. The inoculation method consisting of immersion of the mini-setts in bacterial suspension proved to be more efficient than the inoculation methods of applying the suspension onto the post-sprouting substrate or to the lowest leaf.
RIG-I is an innate sensor of RNA virus infection and its activation induces interferon-stimulated genes (ISGs). In vitro studies using human cells have demonstrated the ability of synthetic RIG-I ...agonists (3pRNA) to inhibit IAV replication. However, in mouse models of IAV the effectiveness of 3pRNA reported to date differs markedly between studies. Myxoma resistance (Mx)1 is an ISG protein which mediates potent anti-IAV activity, however most inbred mouse strains do not express a functional Mx1. Herein, we utilised C57BL/6 mice that do (B6.A2G-Mx1) and do not (B6-WT) express functional Mx1 to assess the ability of prophylactic 3pRNA treatment to induce ISGs and to protect against subsequent IAV infection. In vitro, 3pRNA treatment of primary lung cells from B6-WT and B6.A2G-Mx1 mice resulted in ISG induction however inhibition of IAV infection was more potent in cells from B6.A2G-Mx1 mice. In vivo, a single intravenous injection of 3pRNA resulted in ISG induction in lungs of both B6-WT and B6.A2G-Mx1 mice, however potent and long-lasting protection against subsequent IAV challenge was only observed in B6.A2G-Mx1 mice. Thus, despite broad ISG induction, expression of a functional Mx1 is critical for potent and long-lasting RIG-I agonist-mediated protection in the mouse model of IAV infection.
Highlights • P2X7R antagonisms could be a new advantageous target for the treatment of cancer-induced bone pain. • P2X7R antagonism reduced neuronal responses in the dorsal horn. • P2X7R antagonism ...reduced pain behavior in awake animals, without affecting motor coordination. • P2X7R antagonism has no effect in naïve or sham animals, suggesting a state-dependent effect. • The analgesic effect was not associated with increased expression of P2X7R in the spinal cord.
Both the administration of growth factors and physical therapy such as forced arm use (FAU) are promising approaches to enhance recovery after stroke. We explored the effects of these therapies on ...behavioral recovery and molecular markers of regeneration after experimental ischemia.
Rats were subjected to photothrombotic ischemia: sham (no ischemia), control (ischemia), brain-derived neurotrophic factor (BDNF; ischemia plus BDNF, 20 microg), and FAU (ischemia plus FAU, 1-sleeve plaster cast ipsilateral limb). Animals survived 1 or 6 weeks and underwent behavioral testing (Rotarod, beam balance, adhesive removal, plantar test, neuroscore). After the rats were killed, brain sections were immunostained for semiquantitative analysis of MAP1B, MAP2, synaptophysin, GFAP expression, and quantification of infarct volumes.
Infarct volumes were not different between the groups 1 or 6 weeks after ischemia. BDNF-treated animals had better functional motor recovery (Rotarod, beam balance, neuroscore) compared with all other groups (P<0.05). There was no significant adverse effect of early FAU treatment on motor recovery, although sensorimotor function (adhesive removal test) was impaired (P<0.05). There were no differences between groups as measured by nociception of the left and right forepaw (plantar test). BDNF treatment transiently induced MAP1B expression in the ischemic border zone and synaptophysin expression within the contralateral cortex 6 weeks after ischemia (P<0.05). Both BDNF and FAU reduced astrogliosis compared with controls (P<0.05).
Postischemic intravenous BDNF treatment improves functional motor recovery after photothrombotic stroke and induces widespread neuronal remodeling. Early FAU treatment after stroke does not increase infarct size, impairs sensorimotor function, but leaves motor function unchanged. Postischemic astrogliosis was reduced by both treatments.
Seven adult human cadaveric cervical spines (C2-T1) were biomechanically tested in a programmable testing device.
Compare the effects of incremental single-level fusion at different levels of the ...cervical spine.
Clinical studies have reported degenerative symptomatic disc disease at disc levels adjacent to fusion. No known study has attempted to delineate the effects of single-level fusion at different levels of the cervical spine.
The spines were tested in flexion, extension, right and left lateral bending, and right and left axial rotation for 7 different conditions: harvested and 6 independent single-level fused conditions (i.e., C2-C3, C3-C4, C4-C5, C5-C6, C6-C7, and C7-T1). Segmental motion and global stiffness data were normalized to the harvested condition and compared using a 1-way analysis of variance followed by a SNK test (P < 0.01).
Motion compensation was distributed among the unfused segments with significant compensation at the segments adjacent to fusion. Significant increases occurred at the level above C3-C4 and C4-C5 fusions, and below for C5-C6 and C6-C7 fusions in both flexion and extension.
Increase motion compensation occurred at segments immediately adjacent to a single-level fusion. Significant differences occurred at the level above the fusion site for the C3-C4 and C4-C5 fusion in both flexion and extension. When the lower levels (C5-C6, C6-C7) were fused, a significant amount of increased motion was observed at the levels immediately above and below the fusion. However, greater compensation occurred at the inferior segments than the superior segments for the lower level fusions (C5-C6, C6-C7).
Perfusion and angiographic imaging using intravenous contrast application to evaluate stroke patients is now technically feasible by flat detector CT performed by the angiographic system. The aim of ...this pilot study was to show the feasibility and qualitative comparability of a novel flat detector CT dynamic perfusion and angiographic imaging protocol in comparison with a multimodal stroke MR imaging protocol.
In 12 patients with acute stroke, MR imaging and the novel flat detector CT protocol were performed before endovascular treatment. Perfusion parameter maps (MTT, TTP, CBV, CBF) and MIP/volume-rendering technique images obtained by using both modalities (MR imaging and flat detector CT) were compared.
Comparison of MIP/volume-rendering technique images demonstrated equivalent visibility of the occlusion site. Qualitative comparison of perfusion parameter maps by using ASPECTS revealed high Pearson correlation coefficients for parameters CBF, MTT, and TTP (0.95-0.98), while for CBV, the coefficient was lower (0.49).
We have shown the feasibility of a novel dynamic flat detector CT perfusion and angiographic protocol for the diagnosis and triage of patients with acute ischemic stroke. In a qualitative comparison, the parameter maps and MIP/volume-rendering technique images compared well with MR imaging. In our opinion, this flat detector CT application may substitute for multisection CT imaging in selected patients with acute stroke so that in the future, patients with acute stroke may be directly referred to the angiography suite, thereby avoiding transportation and saving time.
In patients with end-stage renal disease, anemia develops as a result of erythropoietin deficiency, and recombinant human erythropoietin (epoetin) is prescribed to correct the anemia partially. We ...examined the risks and benefits of normalizing the hematocrit in patients with cardiac disease who were undergoing hemodialysis.
We studied 1233 patients with clinical evidence of congestive heart failure or ischemic heart disease who were undergoing hemodialysis: 618 patients were assigned to receive increasing doses of epoetin to achieve and maintain a hematocrit of 42 percent, and 615 were assigned to receive doses of epoetin sufficient to maintain a hematocrit of 30 percent throughout the study. The median duration of treatment was 14 months. The primary end point was the length of time to death or a first nonfatal myocardial infarction.
After 29 months, there were 183 deaths and 19 first nonfatal myocardial infarctions among the patients in the normal-hematocrit group and 150 deaths and 14 nonfatal myocardial infarctions among those in the low-hematocrit group (risk ratio for the normal-hematocrit group as compared with the low-hematocrit group, 1.3; 95 percent confidence interval, 0.9 to 1.9). Although the difference in event-free survival between the two groups did not reach the prespecified statistical stopping boundary, the study was halted. The causes of death in the two groups were similar. The mortality rates decreased with increasing hematocrit values in both groups. The patients in the normal-hematocrit group had a decline in the adequacy of dialysis and received intravenous iron dextran more often than those in the low-hematocrit group.
In patients with clinically evident congestive heart failure or ischemic heart disease who are receiving hemodialysis, administration of epoetin to raise their hematocrit to 42 percent is not recommended.
The potential neuroprotective effect of the granulocyte colony-stimulating factor (G-CSF) after glutamate-induced excitotoxicity in cell culture and after focal cerebral ischemia in rats was studied. ...We hypothesized the existence of the G-CSF receptor (G-CSFR) as a main G-CSF effector on neurons, and immunohistochemistry, immunoblotting, and polymerase chain reaction were performed. The G-CSFR-mediated action was studied by activation of signal transducer(s) and activator(s) of transcription-3 (STAT3) in the periphery of the infarction.
Neuroprotection of various G-CSF concentrations on glutamate-induced excitotoxicity was studied in cell culture. In vivo, ischemia was induced by use of a suture occlusion model of the middle cerebral artery (90-minute occlusion) in the rat. Thirty minutes after the induction of ischemia, the animals (n=12 per group) received G-CSF at 60 microg/kg body wt IV for 90 minutes or vehicle (saline). Infarct volume was calculated on the basis of 2,3,5-triphenyltetrazolium chloride staining 24 hours after ischemia. Expression of the G-CSFR was studied by immunohistochemistry and verified by reverse transcription-polymerase chain reaction and immunoblotting. Expression of STAT3 was determined by immunohistochemistry.
In cell culture, G-CSF exhibited a significant neuroprotective effect after glutamate-induced excitotoxicity (P<0.05). A G-CSF concentration of 10 ng/mL was maximally effective, resulting in a nearly complete protection. In vivo, G-CSF reduced infarct volume to 47% (132.0+/-112.7 mm3 versus 278.9+/-91.6 mm3 P<0.05 in the control group). Immunohistochemistry, Western blotting, and reverse transcription-polymerase chain reaction revealed the existence of G-CSFRs in neurons and glial cells. Animals treated with G-CSF significantly upregulated STAT3 in the periphery of the infarction compared with control animals (P<0.05).
G-CSF achieved a significant neuroprotective effect in cell culture and after intravenous administration after stroke. Increased STAT3 expression in the penumbra of G-CSF-treated rats suggests mediation by G-CSFR.
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