Month: August 2020

In our study, MEAP showed significant anti-nociceptive activities in a dose-dependent manner in the acetic acid-induced writhing tests in mice. In addition, the acetic acid-induced writhing test is commonly used for evaluation of peripheral anti-nociceptive activity of drugs. Acetic acid can indirectly induce the release of endogenous mediators and stimulate the nociceptive neurons that are sensitive to non-steroidal anti-inflammatory drugs. The hot plate test is widely used for evaluating central antinociceptive activities. Results of the hot plate test in our present study indicated that MEAP is not a centrally acting analgesic. Therefore, we presumed that the MEAP exerts peripheral anti-nociceptive activities and performed the formalin test to confirm the hypothesis. The formalin test is a pain model consisting of two different LEE011 phases, which can be separated in time. The early phase is generated peripherally through the activation of nociceptive neurons by the direct action of formalin, and the late phase refers to the inflammatory pain response involving the release of molecules such as prostaglandin, histamine and serotonin. In our present study, the MEAP mainly suppressed pain of the late phase, which suggested that the anti-nociceptive mechanism of MEAP is related to the peripheral anti-nociceptive activity. This result was confirmed by testing MEAP in combination with naloxone. By using the openfield test, a behavior model used to study exploratory and motor activity, we observed no obvious changes of locomotor activity, which provided another piece of solid evidence to support the peripheral anti-nociceptive mechanism of MEAP activity. Cyclooxygenase is the key enzyme in the synthesis of prostaglandins, and COX-2, an inducible enzyme, is responsible for the production of the pro-inflammatory prostaglandins. Many former investigations have shown the important role of COX-2 in the induction of pain and inflammation as well as the analgesic actions of NSAIDs. In addition, COX-2 can induce synthesis of PGE, and the PGE2 released in inflamed tissues sensitizes the terminals of afferent nerve fibers, thereby enhancing nociceptive processing within the spinal cord and brain to evoke hyperalgesia. Thus, one apparently feasible approach against nociception is to suppress the release of COX-2. In our study, MEAP significantly and dose-dependently inhibited expression of COX-2 in the spinal dorsal horns of pain model mice induced by formalin, which indicated that MEAP may be a potent COX-2 inhibitor. Pituitary adenylate cyclase-activating polypeptide is a neuropeptide with multiple roles, including neurotransmitter, neuromodulator and neurotrophic factor. Our recent studies have suggested that PACAP is associated with psychiatric disorders, including schizophrenia.

The chronic loss of brain cortex was reduced by the hEPO+MBs/FUS treatment. These results indicated that MBs/FUS enhanced the hEPO entry even 5 h after I/R, which resulted in neuron protection in both acute and chronic phases. Although stroke itself might alter hEPO delivery, the amount of hEPO entering the infarction area did not produce significant therapeutic effect. As hEPO combined with MBs/FUS, it can result in a significant neuroprotection on both acute and chronic phases. It has been demonstrated that intracerebraventricular administration of hEPO inhibits the I/R-induced brain injury. However, direct injection of hEPO into the brain is not a practical approach to have an appropriate hEPO distribution in the entire infarcted region. In the meanwhile, this kind of interstitial method can result in severe hemorrhages and brain trauma. On the contrary, systemic delivery of hEPO can have a much more uniform distribution of hEPO in the infarcted SB431542 volume but may be limited by the therapeutic time window. In this study, transcranial, noninvasive FUS technology was demonstrated to be a useful modality to transiently open the localized BBB for the targeted delivery of neuroprotectant to treat the ischemic stroke-induced brain injury beyond the conventional therapeutic time window. Brines et al. reported that animals receiving hEPO,3 h after occlusion showed significant reduction of necrosis volume compared with controls. Animals receiving hEPO 6 h after occlusion exhibited a significant decrease in injury volume, but the effect was substantially smaller compared with animals receiving hEPO earlier. Gan et al. reported that EPO exerted significantly neuroprotective effects when administered up to 4 h after I/R in MCAO model, but the effects were significantly diminished and lost when administered 6 h after I/R. In our study, we employed 3VO for 50 min and injected EPO at 5 h after reperfusion and the result showed that there was no significant neuroprotection. These might be due to different stroke models with various occlusion and ischemic duration would produce different levels of impact on the brain. EPO-TAT administered at the onset of post-stroke reperfusion showed the ability across the BBB for neuroprotection. Derivatives of EPO such as CEPO had the neuroprotection ability only within 4 h after occlusion, which is equal to 3 h after reperfusion, in a rat model of focal ischemia. BBB is more permeable for these EPO derivatives within 3 h after reperfusion due to leaky vascularity. Mutant EPO exerted neuroprotective effects up to 4 h after reperfusion but gradually lose its efficacy as time went by. However, the neuroprotective effects were diminished and lost when the mutant EPO was administered 6 h after reperfusion. Ischemia is an acute pathological process and cells die rapidly within first several hours after ischemia. Therefore, neuroprotective drugs must be delivered within their therapeutic window. In this study, we demonstrated that MBs/FUS had the ability to enhance EPO into the brain at 5 h after reperfusion.

In the inflammatory response, transcriptional factors such as nuclear factor-kappaB are activated upon binding of pattern-recognition receptors, proinflammatory cytokine receptors, and antigen receptors. Although NF-kB does not fit the classical definition of an oncogene, it is a powerful activator of the malignant state and regulates the expression of target genes important for cell proliferation, survival, angiogenesis, and tissue repair. Exposure to environmental factors, such as infectious agents, dietary carcinogens, and hormonal imbalances, is thought to lead to injury of the prostate and the development of chronic inflammation. Recent reports showed that Propionibacterium acnes is frequently detected in prostate tissue from patients with prostatitis and prostate cancer, and that the bacterium is associated with acute and chronic prostatic inflammation and might have a role in prostate carcinogenesis. To further investigate the etiologic association between P. acnes and inflammation or carcinogenesis, not only the bacterium but also histologic features of the prostate tissue need to be analyzed in identical histologic sections. The aim of the present study was to locate P. acnes in prostate tissue under light microscopy by enzyme immunohistochemistry. For this purpose, we developed a novel anti-P. acnes monoclonal antibody that reacts with the bacteria in formalin-fixed and paraffin-embedded prostate tissue sections. To evaluate the pathogenic role of this indigenous bacterium in the development of prostate cancer, we examined radical prostatectomy samples obtained from patients with or without prostate cancer by immunohistochemistry with the novel antibody to P. acnes and an antibody to NF-kB, which was used to determine a possible correlation between P. acnes infection and nuclear NF-kB expression in prostate WY 14643 inquirer glands. Furthermore, we analyzed whether P. acnes infection status was associated with prostate cancer risk. Enzyme immunohistochemistry with a novel P.acnes-specific monoclonal antibody enabled us to visualize prostatic P.acnes under a light microscope for histopathologic analysis. In the present study, cocktail immunostaining with both the PAL antibody and anti-NF-kB antibody, followed by morphometric analysis of positive signals on virtual slides, revealed a possible correlation between intraepithelial P. acnes infection and NF-kB activation in prostate glands. A diagnosis of prostatic P. acnes infection needs to be supported by histologic detection of the bacterium in tissue sections, because this indigenous bacterium may cause contamination and tissue invasiveness cannot be evaluated when traditional culture and polymerase chain reaction-based methods are applied. In previous reports, prostatic P. acnes was visualized by fluorescence immunohistochemistry or fluorescence in situ hybridization methods, although a precise histopathologic examination of the prostate lesions cannot be achieved by these methods. In the present study, we used the enzyme immunohistochemistry with the PAL antibody, which reacts with P. acnes with high specificity on routine histologic sections of the formalin-fixed paraffin-embedded prostate tissues.

The overall rationality for their use in IBS has been doubted, because a lack of specific mechanism of action has been confirmed. However, almost all probiotic combinations contained Streptococcus, it is therefore possible that Streptococcus cooperated with other species of probiotics are synergistic in promoting a therapeutic effect in IBS. In this study, PI-IBS mouse after gavaged with mixture of three species, ameliorated visceral sense, intestinal permeability and cytokine profiles. Compared with single species, the Mixture has, to some extent, evident advantages. According to the expression of occludin, the Mixture group was higher than Lactobacillus. In addition, the Mixture group showed decreased expression of IL-17 compared to Bifidobacterium. Based on these results, we could conclude that mixture of three stains was superior to single species. VSL#3, probiotic ‘cocktail’, was reported to be a novel probiotic for the treatment of IBS. In IBS patients with predominant bloating, VSL#3 significantly reduced flatulence scores and retarded colonic transit in contrast to placebo. The comparison between single probiotic and combination probiotic was not reported before, but it turned out that combination was superior to single species in this study. Thus, we have demonstrated the superiority of mixture of three species in barrier protection as well as immunoregulation. In summary, the GDC-0449 literature confirms the benefit of Bifidobacterium and Lactobacillus alone or the combination of the three species on the gut sensation, intestinal permeability in PI-IBS mouse model; the mechanisms supporting these beneficial effects may be upregulation of tight junction proteins and restriction inflammation. Nonertheless, Streptococcus shows either no effect or a favorable effect. Most importantly, we have demonstrated the superiority of mixture of three species over a single one. This study may aid our understanding of the mechanisms underlying probiotic treatments for PI-IBS, which might offer referrences to select appropriate probitic species for IBS patients with different symptoms. The American Stroke Association estimates that stroke accounts for 1 out of every 18 deaths and occurs every 40 seconds in the United States. In the 2012 update, the majority of strokes were ischemic ; 10% were intracerebral hemorrhage; 3% were due to subarachnoid hemorrhage. Ischemic stroke results from the occlusion of a cerebral artery, leading to a blocked cerebral blood flow to certain part of the brain. Two emergent clinical therapies for acute ischemic stroke are: reperfusion of the blood flow and neuroprotection of the injured brain cells. Early reperfusion within 3 h is beneficial to improve the outcome of acute human ischemic stroke. However, late recovery of circulation might cause reperfusion injury, resulting in blood-brain barrier breakdown, or brain edema. Although many animal stroke models have been developed, no single model can fully mimic clinical human stroke because of its heterogeneity. The transient three vessels occlusion method provides a model for the study of ischemia-reperfusion injury. This method can build a stable focal infarction in the brain.

The slight but significantly delayed recovery from hyperglycemia observed in the IH group may indicate that the beta cells after IH treatment become dysfunctional, and that is why the levels of insulin in the IH animals are low. Role of zinc in insulin production Disturbance of zinc homeostasis has been accepted as one of the most crucial signs of diabetes because diabetic patients experience an increased urinary excretion of zinc and zinc is involved in the synthesis, secretion as well as function of insulin. The zinc level in serum as well as in the cytoplasm of beta cells in diabetic patients is low compared to normal levels irrespective of types of diabetes. Despite the general congruence in opinions, some clinical study defies a one-to-one relationship between the zinc level in beta cells and the level of insulin secretion. Our current results show that insulin production decreases as the ARRY-142886 intracellular zinc level decreases after IH challenge; however, mRNA production of insulin does not show any change. This result may indicate IH insults do not influence the beta cells at the transcription level of insulin, but may influence the assembly process in the production line. The human body contains 2–4 g of zinc in total, but the mobile amount of zinc in plasma is very small, i.e. approximately 12– 16 mM. Mainly, zinc is bound to proteins such as metallothionein in cytoplasmic compartments of zinc abundant cells like pancreatic beta cells. Zinc accumulated in the ER and Golgi apparatus in pancreatic beta cells plays a dual function, i.e. anti-oxidative function and insulin production/secretion function. This may be why our IH model is extremely prone to be diabetic. The additional ROS accumulation from our IH challenge should have consumed zinc in the insulin containing vesicles, ER, as well as Golgi apparatus. Some recent studies view zinc as an intracellular second messenger augmenting insulin activity via ZIP which plays a role of sensor. In fact, there is ample evidence showing that increased zinc levels can boost insulin production and secretion. Hence, Myers et al. explained that the increased zinc concentration in the cytoplasm would increase numbers of ZIPs as ZIP7 did in another study, then as a result, increase insulin secretion. They also quoted that this is why oral administration of zinc improves symptoms in type 1 and type 2 diabetic patients. However, the anti-diabetic effect of zinc is still controversial due to a weak dose-effect relationship clinically. Since most zinc is bound with ligands in beta cells, whether the signaling process of zinc to alarm the innate defense system in response to a physiologic challenge by immediate ‘buffering and muffling’ is sufficiently effective remains to be answered. We have not performed a rescue experiment yet; however, it would be very interesting to observe whether zinc administration would recover ZIP8 expression, and in turn recover insulin secretion in our animal model as well as in vitro model. In fact, no reports are available as to whether zinc administration increases ZIP8 concentration in beta cells.