Category: clinically Small Molecule

Acylation has previously been shown to increase intestinal permeability of peptide drugs, but detailed investigations of systematic acyl variations are lacking, which would benefit rational new designs of peptide drugs. The in vitro intestinal translocation studies can be further supplemented by measurements of peptide binding to model lipid membranes in order to investigate the influence of membrane binding of acylated peptides on cellular membrane translocation. Glucagon-like peptide-2 is a 33 amino acid peptide, which is secreted from the human intestine following nutrient intake. Therapeutically, GLP-2 stimulates intestinal growth and is employed in the treatment of inflammatory bowel diseases and short bowel syndrome. The plasma half-life of GLP-2 in humans is limited to a few minutes due to extensive renal clearance and rapid enzymatic degradation by dipeptidyl peptidase-4. Furthermore, GLP-2 is presently administered as subcutaneous injections, which compromises patient comfort and compliance, in particular for chronic diseases like Crohn’s. It would be highly beneficial to enable oral administration, and the combined effects of prolonged circulation time, improved enzymatic stability and intestinal permeability may render acylated GLP-2 a suitable candidate for oral drug delivery. Currently, however, there are no reports on the intestinal permeability or oral drug delivery potential of acylated GLP-2. In the present study we synthesized and characterized acylated analogues of GLP-2, with systematically increasing acyl chain length, in order to investigate the effect of the acyl chain on membrane interaction and in vitro intestinal permeability. This was achieved by combining investigations of the interaction with lipid membranes and translocation across an intestinal cell model, as outlined in fig. 1. We hypothesize that the acylation chain length can be optimized for translocation across the intestinal barrier, i.e. a moderate interaction with the lipid cell membrane is beneficial for translocation, whereas a stronger interaction may impair translocation. Acylation is expected to Enzalutamide 915087-33-1 confer membrane affinity to GLP2, as the native peptide is not membrane active. In this regard, GLP-2 was employed as a model peptide, however, the results may be applicable for development of a rational acylation strategy for other peptide drugs. Absorption enhancers are often employed to increase oral peptide absorption, which makes it interesting to investigate how these affect the translocation of acylated peptides. In the present study we included two enhancers with different enhancing mechanism, in order to investigate the effect of the enhancing mechanism. Ethylene glycol-bis-N, N, N, Ntetraacetic acid is a paracellular enhancer which increases transport between the cells by opening of the tight junctions, and sodium dodecyl sulfate is a transcellular enhancer which increases transport through the cells at low concentrations, predominantly by fluidizing the cell membrane. We hypothesize that the effect of paracellular enhancers will not be influenced by acylation, whereas the effect of transcellular enhancers that directly interact with the cell membrane may depend on the peptide-membrane interaction, through altered membrane affinity and/or dynamics of membrane insertion. For EGTA the increase is similar for peptide and analogues, and the dependence on acyl chain length is retained, whereas for SDS the increase is greater for the c16-acylation. These results support the hypothesis that the fluidization of cell membranes caused by SDS are beneficial for the long chain acylation, possibly due to altered membrane insertion.

The effects of MF exposure on DNA in situ in a cell type-specific manner, we carried out further experiments using a very similar experimental setup and mode of evaluation as described previously. The following questions were addressed: Does MF exposure with much lower flux density than 1.5 mT also result in significant effects on DNA? If this is the case, are all cells or only specific cell types affected ? How do cells that are highly involved in iron storage react to MF exposure? Experiments were carried out on mice with MF flux density of 0.1 mT at 50 Hz as this value represents the current exposure limit for the general population in most European countries. In order to study dose-dependent effects, we also exposed mice to 1.0 mT at 50 Hz. Alongside the brain and kidney, we also analyzed liver cells strongly involved in iron storage. There are over 250 million people in the world with type 1 and 2 diabetes mellitus. Neuropathy is one of the most common complications of diabetes mellitus and leads to increasingly high morbidity and mortality, resulting in a huge economic burden for diabetes care. Diabetic neuropathy is a heterogeneous condition containing symmetrical neuropathies and focal neuropathies, presenting diverse clinical manifestations. Of all the neuropathies in diabetes, chronic diabetic peripheral neuropathy is the commonest. Of all the symptoms in DPN, pain is the most distressing and is the main factor that prompts the patients to seek medical advice. One-third of diabetic patients have symptoms of neuropathic pain according to a recent community-based study and up to 15–20% of patients with DPN may experience painful symptoms. Therefore, a high proportion of patients are suffering from neuropathic pain as well as the relative depression, anxiety and sleep deprivation. The management of neuropathic pain in diabetes still remains challenging mainly due to its various clinical features, wide spectrum severity and different Y-27632 dihydrochloride inquirer distribution involved. Descriptions of pain can be burning, prickling, lancinating, shooting, cramping, aching, and also contact hypersensitivity and “dead feeling” in their legs. The severity may range from mild symptoms in one toe or two to continuous painful symptoms involving both legs and may even extend to the upper limbs. The extent involved may be focal or diffuse. One additional factor that contributes to the treatment dilemma of neuropathic pain is the varied response to the currently different treatments. The diverse manifestations of neuropathic pain in diabetes and various responses to current interventions imply that a number of mechanisms could contribute. Therefore, the management of painful DPN may not be one single intervention and a series of factors should be taken into consideration, one of which, as Vinik, A. and his colleagues put it in one guideline, may be the distribution of pain. According to our clinical experience with management of painful diabetic neuropathy, features and severity of pain may change during the course of diabetic neuropathy while the distribution of pain is relatively invariable, which may be of some value for patient selection for surgical decompression. Thus we carry out this retrospective study to investigate the effects of surgical decompression on the outcome of painful diabetic patients and discuss the role which pain distribution plays in the management of painful diabetic neuropathy as well as the underlying mechanism involved. Aside from traditional management including glucose control, lifestyle modification and pharmacological treatment, surgical decompression is recommended for pain relief in the recent reports based on the “double crush” hypothesis.

But the recruitment strategy of selecting loyal patients is likely to reduce this chance. Fourth, the compliance of insulin medication was not examined. In conclusion, no cancer signal with insulin glargine was found in this carefully characterized clinical cohort with diabetes. While our study is limited in size, we avoided potential for major distortions by implementing a new user, active comparator study design. Our study thus adds to the evidence that insulin glargine does not increase the risk for any cancer outcomes when compared with its main treatment alternative, NPH insulin. Chemotherapy is still the main clinical treatment for cancer. Most of the chemotherapeutic drugs induce serious multi-drug resistance and a series of side effects, e.g., fatigue, muscle and joint pain, impaired immune responses, anemia, neutropenia and thrombocytopenia. Therefore, searching for novel anti-tumor agents from natural products with fewer adverse effects is highly important. The use of medicinal mushrooms in the fight against cancer has been known for a very long time in Korea, China, Japan, Russia, USA and Crizotinib 877399-52-5 Canada. Mushrooms produce a variety of complex, lowmolecular-weight compounds with diverse chemical compositions, such as phenolic compounds, polyketides, triterpenoids and steroids. Many have shown direct beneficial effects on cancer development by interfering with specific transduction pathways. Mushroom Pyropolyporus fomentarius Teng, also called Fomes fomentarius, is a fungus of the Polyporaceae family that acts a parasite on beech and birch trees ; it has worldwide distribution. P. fomentarius has wideranging uses, including medicinal use. Many bioactive substances were isolated from the P. fomentarius petroleum ether fraction, such as b-sitosterol, 5a,8a-epidioxy-ergosta-6,22-dien-3bol, ergosta-7,22-dien-3b-ol, ergosta-7,22-dien-3-palmitate, Stearic acid, Palmitic acid, Ergosta-7,22-dien-3-one, ergosta7,22-dien-3-one, dimethyl acetal and sterols. However, there no studies have demonstrated its anti-tumor activities and the underlying mechanism. Many traditional herbal medicines have shown antiproliferative effects on the S180 cell line or cytotoxic activities on the S180-bearing mouse model, alone or combined with cyclophosphamide. The present work has demonstrated the significant anti-tumor activity of the PFPE both in vitro and in vivo. One of the anti-tumor activity chemotherapeutic targets is cytotoxicity. Most clinically used anti-tumor agents possess significant cytotoxic activity in cell culture systems. In our paper, PFPE was toxic to S180 cells at 240 and 480 mg/ml in time-dosedependent manners. However, at the concentration of 120 mg/ml, there was no proliferation inhibition effect with the prolonged incubation time. The dose response phenomenon and low dose stimulation have been previously reported for other drugs. The lower concentration might have other functions, such as anti-inflammation, anti-virus, etc., and the underlying mechanisms require further exploration. Moreover, PFPE had no or little cytotoxicity in HEK-293 cells. Thus, the present study suggests that PFPE has a potential application as a natural anti-tumor agent. Defects in apoptosis are the critical step in the resistance to therapy in many types of cancers. Thus, apoptotic pathways are relevant targets in cancer therapies. Previous studies have shown that apoptosis is an important mechanism through which various anticancer agents exert anticancer effects. In the present study, we aimed to determine whether apoptosis was induced in S180 cells along with the PFPE exposure. As evidenced by Annexin V-FITC/PI double staining, we found that the proportion of early and late apoptotic cells increased significantly after the PFPE treatment.

As shown in Figure 4, PFPE caused obvious DNA fragmentation in S180 cells, which is also a typical biochemical feature of apoptosis. The data in this study suggest that the PFPE could induce apoptosis in S180 cells, and the cytotoxic effects were associated with apoptosis, implying that the extract has great potential in anti-cancer drug screening. Mitochondria play a critical role in cell apoptosis triggered by many stimuli. Loss of MMP is an early event in apoptosis. The MMP, as detected by flow cytometry with Rh123, significantly decreased immediately after 24 h PFPE treatment. Electron leakage from the mitochondrial respiratory chain may react with molecular oxygen, resulting in the formation of superoxide, which is subsequently converted to ROS. Moreover, excessive ROS may cause oxidative damage to lipids, proteins, and DNA, leading to cell death. As seen from Figure 6, compared with the control group, the generation of intracellular ROS dramatically increased in S180 cells. These results suggest that the decreased MMP and elevated ROS may both related to cell apoptosis after the PFPE treatment, and the PFPE-induced cell apoptosis might be mitochondria dependent. Consistent with in vitro findings, the in vivo study provides information that the PFPE significantly reduced the S180 sarcoma weight at the indicated dose, showing its specific role in anticancer therapy. In tumor immunotherapy, the occurrence, growth of tumor and immune state has a very important relationship. Spleen and thymus are important immunological organs. The spleen index and thymus index could reflect the immune Adriamycin function of spleen and thymus. According to other reports, CTX caused atrophy of spleen and thymus and influenced the spleen. We got similar results in the in vivo experiment in CTX treated group. However, at low concentration group, PFPE did not induce such changes. As illustrated in our work, PFPE could efficiently inhibit tumor growth and also has lower immune organ toxicity. Via GC–MS analysis, various possible chemical components were detected in PFPE. n-Hexadecanoic acid has antitumor activity to human leukemic cells as well as murine cells. Hexadecanoic acid has been reported to induce NF-kB activation in HaCaT keratinocytes, which is an important pathway involved in cancer development. Moreover, fatty acids exhibit cytotoxicity against HeLa cells and retard tumor growth. The essential oil, with n-Hexadecanoic acid and octadecanoic acid as the main components, showed significant cytotoxicity against oral cancer, breast cancer and small cell lung cancer. Steroidal compounds had a potent inhibition effect against various cancer cells. Therefore, the anti-tumor effect of PFPE both in vitro and vivo may be closely related to the specific efficacy of such components, functioning either alone or together, which needs further confirmation. In conclusion, this work provided evidence that the PFPE inhibits the proliferation of S180 cells in vitro through inducing apoptosis, while the PFPE significantly reduced the weight of S180 sarcoma in vivo. Apoptosis induction has become a new therapeutic target in cancer research, and the results in our paper indicate that the PFPE may have a potential application as a natural anti-tumor agent. Future research should investigate the specific bioactive compounds and then try to explore the molecular mechanisms of the PFPE in cancer therapy. The cyclic nucleotides, cyclic adenosine monophosphate and cyclic guanosine monophosphate are intracellular second messengers with diverse regulatory functions in both unicellular and multicellular organisms. Hence there are an extreme variety and large number of isoforms of these nucleotidyl cyclases.

First, in line with previous reports in SLE patients,, our data indicate that ApoCell-phagocytosis is deficient in several MDM preparations that were derived from SS and SLE patients by cultivation in the absence of patients’ sera, a fact that may at least partly support a model of intrinsically defective monocyte in these disorders. Alternatively, the impaired capacity of phagocytes of SS and SLE patients for uptake of prey may be viewed as a result of cellular activation taking place in vivo, a notion probably also implied by the significant correlation between the findings in the various phagocytosis assays and the activity indices of these diseases. Finally, the notably low capacity of SS and SLE patients for phagocytosis of particulate targets by blood-borne monocytes and MDM that was observed may reflect an overall dysfunction of phagocytes in the proper engulfment of certain preys in these disorders. Interestingly, deficient phagocytosis of particulate targets is also reported in primary biliary cirrhosis, a disease with striking clinicopathologic similarities to SS. On the other hand, it should be noticed that in sharp contrast to the above deficient phagocytic capacities, the bloodborne monocytes of SS and SLE were found remarkably hyperfunctional in the phagocytosis of necrotic cell debris. The addition of healthy serum was found to facilitate significantly the ingestion of apoptotic cells by blood-borne monocytes derived from healthy individuals, as well as from SLE or SS patients. These findings are in good agreement with the previously observed capacity of sera from healthy individuals to restore the phagocytic ability of macrophages from SLE patients. In addition, ApoCell-phagocytosis by healthy monocytes was presently shown to be severely impaired following the substitution of HBD sera by sera derived from SS and SLE patients. Consistent with previous observations, the sera from approximately 80% of SLE patients studied were not as efficient as healthy sera in supporting of ApoCell-phagocytosis by normal blood-borne monocytes. In this study, we present first evidence that such incapacity is also manifested by the vast majority of sera from SS patients studied. The precise nature of this aberration in SS and SLE patients is unclear. Normally, several serum proteins attach to apoptotic cells and induce the deposition of C3 and its degradation products C3b and iC3b, thus enhancing efferocytosis by phagocytes via recognition by complement receptors CR3 and CR4. In this context, mice lacking such bridging molecules, such as MFG-e8, Mer or C1q are reported to develop lupus-like manifestations GW-572016 associated with inefficient removal of apoptotic cells. In line to these observations, several quantitative and functional aberrations of the above opsonins have been described in SLE and SS patients, including hypocomplementemia, which is considered as one of the major immunological markers and of key clinical importance for both disorders.