Tolerogenic dendritic cells produced by lentiviral‐mediated CD40‐ and interleukin‐23p19‐specific shRNA can ameliorate experimental autoimmune encephalomyelitis by suppressing T helper type 17 cells

Down‐regulation of soluble or membrane‐bound co‐stimulatory molecules by RNAi in dendritic cells can prevent the activation of immune responses. Therefore, this study was designed to evaluate the therapeutic efficacy of bone marrow‐derived DCs (BMDCs) transduced with lentiviral vectors to permanently expressed shRNA specific for CD40 (CD40LV‐DCs) and/or p19 subunit of interleukin (IL)‐23 (p19LV‐DCs) mRNAs in experimental autoimmune encephalomyelitis (EAE). In‐vitro studies showed that double‐transduced BMDCs (CD40⁺p19LV‐DCs) resemble tolerogenic DCs due to profound down‐regulation of CD40, lower expression of proinflammatory cytokines (IL‐6 and IL‐12), increased IL‐10 production and stronger stimulation of myelin oligodendrocyte glycoprotein (MOG)_35–55‐specific T cells for production of IL‐10 compared with CD40LV‐DCs, p19LV‐DCs and BMDCs transduced with control lentiviral vector (CoLV‐DCs). Moreover, injection of transduced CD40⁺p19LV‐ BMDCs in EAE mice resulted in more reduction in clinical score, significant reduction in IL‐17 or increased production of IL‐10 by mononuclear cells derived from the lymph nodes or spinal cord compared with CoLV‐DCs‐treated EAE mice. In conclusion, simultaneous knock‐down of CD40 and IL‐23 production by BMDCs may represent a promising therapeutic tool for the treatment of IL‐17‐dependent autoimmune diseases, including multiple sclerosis.     

A Randomized Clinical Trial of Insulin Glargine and Aspart,Compared to NPH and Regular Insulin in Children with Type 1 Diabetes Mellitus

Objective

Appropriate treatment of patients with Type 1 diabetes mellitus (T1DM) is necessary to avoid further complications. This study was performed to compare the efficacy of insulin Glargine and Aspart with NPH insulin and regular insulin regimen in a group of children with T1DM.

Methods

Forty patients with T1DM were enrolled in this study. During run-in, all subjects were treated with conventional therapy consisting of twice-daily NPH and thrice-daily regular. Following randomization, 20 subjects received Glargine and Aspart and 20 subjects received NPH and Regular insulin.

Findings

Mean HbA1c was 8.8% and 8.6% at first and 8.4% and 8.2% at the end of study for subjects randomized initially to Glargine and Aspart and for those randomized to NPH and Regular, respectively (P>0.05). Mean fasting blood glucose (FBS) of the subjects randomized initially to Glargine and Aspart was 217±101 mg/dL, with no significant difference to 196±75 mg/dL for those randomized to NPH and Regular (P=0.48). This was also true at the end of the study. The difference in total cholesterol and triglyceride between the two groups in the beginning of study and at the end did not show any significance.

Conclusion

The current study showed no significant difference in glycemic control [Glycated hemoglobin (HbA1c) and FBS] and lipid profile (total cholesterol and triglyceride) between two regimes.     

Probing chelation motifs in HIV integrase inhibitors

A series of HIV integrase (HIV-1 IN) inhibitors were synthesized to evaluate the role of the metal-binding group (MBG) in this class of metalloenzyme inhibitors. A total of 21 different raltegravir-chelator derivative (RCD) compounds were prepared that differed only in the nature of the MBG. These IN strand-transfer inhibitors (INSTIs) were evaluated in vitro in cell-free enzyme activity assays, and the in vitro results were further validated in cell culture experiments. All of the active compounds showed selective inhibition of the strand-transfer reaction over 3'-processing, suggesting a common mode of action with raltegravir. The results of the in vitro activity suggest that the nature of the MBG donor atoms, the overall MBG structure, and the specific arrangement of the MBG donor atom triad are essential for obtaining maximal HIV-1 IN inhibition. At least two compounds (RCD-4, RCD-5) containing a hydroxypyrone MBG were found to display superior strand-transfer inhibition when compared to an abbreviated analogue of raltegravir (RCD-1). By isolating and examining the role of the MBG in a series of INSTIs, we have identified a scaffold (hydroxypyrones) that may provide access to a unique class of HIV-1 IN inhibitors, and may help overcome rising raltegravir resistance.     

Brain Inconspicuous Effect by Local Sinusoidal Extremely Low Frequency Magnetic Exposure Based on Wavelet Packet Analysis: Innovation in Online Passive Neurofeedback Therapy by the Neuro-LSELF System

Neurofeedback (NF) is a training approach that aims to reinforce brain activity by using the information of human electroencephalogram (EEG) rhythms as a feedback. In addition, some studies have reported Extremely Low Frequency (0–300 Hz, intensity < 500 µT) Magnetic Field (ELF MF) effects upon the EEG and its rhythms. The purpose of this study is to determine if an approach that combines the effects of Local Sinusoidal Extremely Low Frequency Magnetic Fields (LSELF MF) with NF yields higher performance on desired NF goals. The NF protocol used in this study consisted of enhancement of the beta rhythm and inhibition of theta and high beta rhythms in exposed and sham groups for the purpose of improving attention. Twenty-four healthy subjects of at least average intelligence attended 10 sessions of NF training. Sixteen of them were exposed to 45 Hz sinusoidal ELF (360 µT) at F3 to lead to the desired LSELF MF effects on Cz. Wavelet packet analysis was used for the detection of changes in EEG rhythms. Results suggest that, compared to sham exposure, LSELF magnetic waves can significantly affect and modulate brainwaves according to this new neurofeedback approach. In comparison to sham exposure, improved ability to attend (as measured by a decrease in the theta-to-beta ratio) was observed when LSELF MF was combined with NF (p < .05). The hypothesis that LSELF MF can affect the theta-to-beta ratio was confirmed. These effects occurred after approximately 10 min of each NF procedure. This study aimed to pilot a new NF system known as the Neuro-ELF system, a method that may allow for more effective control of brainwave activity. However, we suggest that the effects of LSELF-NF require further research.     

Interfacial fracture toughness of different resin cements bonded to a lithium disilicate glass ceramic

Objectives

To evaluate the effect of HF acid etching and silane treatment on the interfacial fracture toughness of a self-adhesive and two conventional resin-based cements bonded to a lithium disilicate glass ceramic.

Methods

Lithium disilicate glass ceramic discs were prepared with two different surface preparations consisting of gritblasted with aluminium oxide, and gritblasted and etched with hydrofluoric acid. Ceramic surfaces with a chevron shaped circular hole were treated by an optimized silane treatment followed by an unfilled resin and then three different resin cements (Variolink II, Panavia F2, and Multilink Sprint). Specimens were kept in distilled water at 37 °C for 24 h and then subjected to thermocycling. The interfacial fracture toughness was measured and mode of failures was also examined. Data were analysed using analysis of variance followed by T-test analysis.

Results

No statistically significant difference in the mean fracture toughness values between the gritblasted and gritblasted and etched surfaces for Variolink II resin cement was found (P > 0.05). For the gritblasted ceramic surfaces, no significant difference in the mean fracture toughness values between Panavia F2 and Variolink II was observed (P > 0.05). For the gritblasted and etched ceramic surfaces, a significantly higher fracture toughness for Panavia F2 than the other cements was found (P < 0.05).

Conclusions

The interfacial fracture toughness for the lithium disilicate glass ceramic system was affected by the surface treatment and the type of luting agent. Dual-cured resin cements demonstrated a better bonding efficacy to the lithium disilicate glass ceramic compared to the self-adhesive resin cement.

Clinical significance

The lithium disilicate glass ceramic surfaces should be gritblasted and etched to get the best bond when used with Panavia F2 and Multilink Sprint resin cements, whereas for the Variolink II only gritblasting is required. The best bond overall is achieved with Panavia F2.     

Synthesis of fluorene and/or benzophenone O-oxime ethers containing amino acid residues and study of their cardiovascular and antibacterial effects

The syntheses and biological studies of O-oxime ethers having α-amino acid residues as new analogs of IPS-339 have been described. In this synthesis, the reaction of fluorene and/or benzophenone O-oxime with epichlorohydrin or epibromohydrin afforded the corresponding O-oxime ether adducts. The N-alkylation of amino acid with O-oxime ether adducts led to synthesis of new analogs of IPS-339. The products were examined for their cardiovascular property. It was demonstrated that 2-(3-(9H-fluoren-9-ylideneaminooxy)-2-hydroxypropylamino)-3-methyl-butanoic acid as the most potent compound substantially reduces the heart rate of dogs. Compounds were also evaluated for their in vitro antibacterial activity against some Gram-negative and Gram-positive bacteria. The antibacterial screening proved the considerable antibacterial activity against both groups of bacteria. The docking analysis demonstrated the appropriate fitting of 2-(3-(9H-fluoren-9-ylideneaminooxy)-2-hydroxy-propylamino)-3-methyl-butanoic acid in human β₂-adrenergic receptor active site. Potential drug toxicity for some active compounds has also been predicted.     

Protein expression of various hepatic uridine 5′‐diphosphate glucuronosyltransferase (UGT) enzymes and their inter‐correlations: a meta‐analysis

Avoiding cytochrome P450 (CYP) related drug interactions in the development of new drug candidates means that glucuronidation by uridine 5′‐diphosphate glucuronosyltransferase (UGT) enzymes is expected to become a more prominent pathway in the metabolism of new drug candidates designed by pharmaceutical companies. Therefore, determining the abundance and activity of these enzymes is of value in the process of scaling in vitro data to in vivo metabolic parameters. Many of the studies involving the measurement of UGTs were conducted with too few samples, which did not provide a good indication of population values and the level of variability. Meta‐analysis is used in the current study to combine all reported values (eight studies that used LC‐MS isotope‐labelled standard targeted quantitative methods), detect inconsistencies between the various datasets and describe correlations of expression between the quantified UGT enzymes. Some heterogeneity was observed between studies, especially in the UGT1A4, 2B7 and 2B10 datasets. However, in the absence of information on the inter‐laboratory consistency of assays, it is difficult to assign these differences to the heterogeneity of the samples. Large inter‐individual variability was observed in the collated data across this family of enzymes. Positive correlations between the expression levels of certain UGT enzymes were found in the collated data. These included the pairs: UGT1A4/2B4 (r_s = 0.71, p < 0.0001, n = 82), UGT2B4/2B15 (r_s = 0.63, p < 0.0001, n = 83), UGT2B7/2B15 (r_s = 0.81, p < 0.0001, n = 99). These correlations can be explained by common regulatory mechanisms involved in the expression of these proteins. Copyright © 2014 John Wiley & Sons, Ltd.     

Inborn errors of metabolism underlying primary immunodeficiencies

A number of inborn errors of metabolism (IEM) have been shown to result in predominantly immunologic phenotypes, manifesting in part as inborn errors of immunity. These phenotypes are mostly caused by defects that affect the (i) quality or quantity of essential structural building blocks (e.g., nucleic acids, and amino acids), (ii) cellular energy economy (e.g., glucose metabolism), (iii) post-translational protein modification (e.g., glycosylation) or (iv) mitochondrial function. Presenting as multisystemic defects, they also affect innate or adaptive immunity, or both, and display various types of immune dysregulation. Specific and potentially curative therapies are available for some of these diseases, whereas targeted treatments capable of inducing clinical remission are available for others. We will herein review the pathogenesis, diagnosis, and treatment of primary immunodeficiencies (PIDs) due to underlying metabolic disorders.