In-vitro effects of Mycobacterium bovis BCG-lysate and its derived heat shock proteins on cytokines secretion by blood mononuclear cells of rheumatoid arthritis patients in comparison with healthy controls

BACKGROUND: Several studies have shown that heat shock protein (HSP)-reactive T cells have an immunoregulatory phenotype indicating that HSPs are able to trigger immunoregulatory pathways, which can suppress immune responses that occur in human inflammatory diseases, such as rheumatoid arthritis (RA). Mycobacterium bovis strain Bacillus Calmette-Guérin (BCG) is rich of HSPs which could be good resources of these regulatory proteins for modulation of immune response. PURPOSES: To study the effects of BCG-lysate and BCG-derived HSPs on secretion of T regulatory cytokines by PBMCs of RA patients in comparison with healthy controls. METHODS: BCG was heat killed and sonicated to have BCG-lysate. BCG-derived HSP65/HSP70 were detected by immunoblotting and purified by preparative SDS-PAGE. PBMCs of 18 RA patients/16 controls collected by Ficoll-paque were stimulated with BCG-lysate/BCG-derived HSP-65/HSP-70. Supernatant of stimulated PBMCs was aspirated for measuring TGF-beta, IL-10, IL-4 and IFN-gamma with sandwich ELISA. RESULTS: BCG-lysate augmented the amounts of all the mentioned cytokines as dose dependent significantly. The level of TGF-beta in controls was higher than patients (P<0.05). HSP65 and HSP70 increased TGF-beta, IL-10 as dose dependent significantly. HSP65, but not HSP70, increased IL-4. HSP65 did not increase IFN-gamma but HSP70 augmented IFN-gamma significantly. BCG-lysate increased IFN-gamma and IL-4 in RA patients more than healthy controls (P<0.05). CONCLUSION: Although BCG is able to provoke T helper 1 cell mediated immunity, its HSP proteins are able to trigger T regulatory cytokines. Healthy controls were under stronger immune regulations.     

Alpha-2 agonists decrease expression of morphine-induced conditioned place preference

Opioidergic system can interact with different transmission such as dopaminergic and adrenergic system. It has been shown that alpha-adrenergic system is involved in some effects of opioid including reward. In this study, alpha-2 agonists were used before testing on day 6 to evaluate their effects on the expression of morphine-induced conditioned place preference (CPP). Our results showed that intraperitoneal (i.p.) injections of morphine (5 mg/kg) induced CPP. Administration of alpha(2)-agonists clonidine (0.01, 0.02 and 0.04 mg/kg, i.p.), tizanidine (0.1, 0.2 and 0.4 mg/kg, i.p.) and xylazine (2.5, 5 and 10 mg/kg, i.p.) decreases the expression of morphine-induced CPP. Yohimbine (0.5 mg/kg, i.p.) reversed the inhibitory effect of alpha(2)-agonists. The comparison of potency of inhibitory effect of three agonists showed that ID(50) values for clonidine, tizanidine and xylazine were 0.013, 0.32 and 1.86 mg/kg respectively. Therefore, it is concluded that alpha(2)-agonists decrease the morphine-induced CPP in mice and clonidine is more potent than tizanidine and xylazine. The relative potency of clonidine with respect to tizanidine and xylazine was 30 and 180 respectively.     

Preparation,statistical optimisation and in vitro characterisation of poly (3-hydroxybutyrate-co-3-hydroxyvalerate)/poly (lactic-co-glycolic acid) blend nanoparticles for prolonged delivery of teriparatide

The purpose of this study was the preparation, optimisation and in vitro characterisation of PHBV and PLGA blend nanoparticles (NPs) for prolonged delivery of Teriparatide. Double emulsion solvent evaporation technique was employed for the fabrication of NPs. The nanoformulation was optimised using the Box–Behnken methodology. The morphological properties of NPs were assessed by both SEM and transmission electron microscopy (TEM). Encapsulation of Teriparatide within the NPs and lacking of chemical bonds between drug and copolymers were proved by XRPD, FTIR and DSC. The structural stability of Teriparatide after processing was confirmed by fluorescence spectrometry. The average size of optimised NPs was 250.0?nm with entrapment efficiency (EE) of 89.5% and drug loading (DL) of 5.0%. Teriparatide release from optimised NPs led to 64.4% release over 30 days and it showed a diffusion-based mechanism. Based on the favourable results, PHBV/PLGA blend NPs could be a promising candidate for designing a controlled release formulation of Teriparatide.     

Determination of a CD4+CD25−FoxP3+ T cells subset in tumor-draining lymph nodes of colorectal cancer secreting IL-2 and IFN-γ

CD4⁺CD25⁻FoxP3⁺ cells are a newly recognized subset of T cells which was first reported in autoimmune diseases. In our previous study, this subset was detected in tumor-draining lymph nodes (TDLNs) of patients with breast cancer. As little is known about their function in TDLNs of cancer patients, in this study, their frequency as well as their ability to produce interleukin (IL)-2, IL-10, or interferon (IFN)-γ were investigated in TDLNs of colorectal cancer (CRC) patients. Mononuclear cells were isolated from lymph nodes of 13 patients with CRC using Ficoll-Hypaque gradient centrifugation. Cells were stimulated in vitro and stained with CD25, CD4, FoxP3, IFN-γ, IL-10, and IL-2 or isotype matched antibodies and subjected to flow cytometry. The frequency of CD4⁺CD25⁻FoxP3⁺CD127^dim/− cells was significantly lower than CD4⁺CD25⁺FoxP3⁺CD127^dim/− population in TDLNs of CRC patients. The percentage of CD127^dim/− cells and also the MFI of FoxP3 expression was significantly lower in CD4⁺CD25⁻FoxP3⁺ in comparison with CD4⁺CD25⁺FoxP3⁺ population. Moreover, CD4⁺CD25⁻FoxP3⁺ cells contained higher percentages of IL-2- and IFN-γ-producing cells than CD4⁺CD25⁺FoxP3⁺ subpopulation. But, no difference was seen between two subsets in terms of IL-10 production. CD4⁺CD25⁻FoxP3⁺ cells in TDLNs of CRC patients had lower suppressive and higher effector properties in comparison with CD4⁺CD25⁺FoxP3⁺ conventional regulatory T cells.

     

Electromechanical coupling, uncoupling, and ventricular function in patients with bundle branch block: a tissue-Doppler echocardiographic study

BACKGROUND: Left bundle branch block (LBBB) is associated with impaired left ventricular (LV) function and increased morbidity and mortality, especially in patients with structural heart diseases. The mechanisms are poorly understood. Subjects and METHODS: Subjects with isolated LBBB (n=20), right bundle branch block (RBBB, n=20), and controls (C, n=20) were studied with standard two-dimensional (2D), and color-encoded tissue-Doppler echocardiography (TDE). Inter- and intraventricular systolic and diastolic coordination were assessed from the TDE velocity profiles. LV function was assessed by 2D echocardiography, by TDE-derived peak systolic velocities, and the atrioventricular (AV) plane displacement. RESULTS: Subjects with LBBB had longer electromechanical delays and longer isovolumic relaxation times than did the C and RBBB groups (P <0.001). For the LBBB subjects compared with the RBBB and C groups, ejection times were shorter, peak systolic velocities and AV plane displacements were lower, they had larger LV end-systolic volumes and lower LV ejection fraction (all P <0.001), and the atrial contribution to A-V plane displacement was higher (P <0.01). There were no differences in diastolic or filling times among the groups. CONCLUSIONS: In patients with LBBB, delayed regional electromechanical coupling and uncoupling leads to generalized intra- and interventricular asynchrony, thereby explaining the depressed regional and global LV functions. Assessment of the electromechanical coupling and uncoupling processes and their consequences on cardiac function in patients with BBB and structural heart diseases may be possible using TDE.     

Do hypertensive individuals have enlarged aortic root diameters? Insights from studying the various subtypes of hypertension

BACKGROUND: Aortic root diameter (AoD) increases with aging and is related to body size. AoD is also presumed to increase in hypertension. In prior studies, however, after adjusting for age and body size, AoD did not differ between hypertensive and normotensive (NT) individuals. Hypertension is a heterogeneous condition with various subtypes that differ in pathophysiology and age distribution. We assessed whether AoD differs among subjects with the various subtypes of hypertension and nonhypertensive individuals. METHODS: In 1,256 volunteers aged 30-79 years (48% women, 48% hypertensive; all untreated), AoD was measured at the sinuses of Valsalva with transthoracic echocardiography. Using cutoff values based on the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure, subjects were identified as NT (23%), or prehypertensive (PH, 29%), or as having isolated diastolic (IDH, 6%), isolated systolic (ISH, 12%), or systolic-diastolic (SDH, 30%) hypertension. Groups were compared using analysis of variance with Bonferroni's correction. RESULTS: AoD increased with age and body surface area (BSA) in both men (r = 0.25 and 0.19, respectively) and women (r = 0.30 and 0.22, respectively) (all P < 0.0001). In men, those identified as having IDH, ISH, and SDH each had a 6% larger AoD than NT individuals (all P < 0.05). In women, those identified with ISH and SDH had a 10 and 8% larger AoD than NT individuals, respectively (all P < 0.05). In both sexes, after indexing to BSA, only ISH individuals exhibited larger AoD compared with NT individuals (both P < 0.05). But, with further adjustment for age, these differences were no longer observed. CONCLUSIONS: Even when the subtypes of hypertension are examined separately, age and BSA, not hypertension status, account for the AoD differences between NT and hypertensive subjects.     

Perspectives of Cardiac Care Unit Nursing Staff about Developing Hospice Services in Iran for Terminally ill Cardiovascular Patients: A Qualitative Study

Introduction:

The present study was conducted aiming to determine the points of view of cardiac care units’ nursing staff about designing and providing Hospice services in Iran for cardiovascular patients in the final stages of life.

Materials and Methods:

In this qualitative study, the perspectives of 16 Cardiac Care Unit (CCU) nurses selected purposefully among hospitals of Tabriz-Iran University of Medical Sciences were investigated using semi-structured interviews and were analyzed in content analysis method.

Results:

33 themes were finally extracted. Some nurses were for and some were against designing and providing Hospice services in Iran. The main reasons identified for supporting this plan included: Possibility of designing and providing these services consistent with high ethical values of Iranian society; approval of authorities due to increasing the load of chronic diseases and aged population; need of families due to the problems in taking care of patients and life concerns; better pain relief and respectful death; decrease of costs as a result of lower usage of diagnostic-therapeutic services, less use of expensive facilities and drugs, and better usage of hospital beds.

Conclusion:

Growing load of chronic diseases has made the need for Hospice as a necessary issue in Iran. In order to provide these services, studying the viewpoints of health service providers is inevitable. Therefore using and applying the results of this study in planning and policy making about designing and providing these services in Iran for cardiovascular patients in their final stages of lives could be helpful.     

Monoclonal antibody conjugated magnetic nanoparticles could target MUC‐1‐positive cells in vitro but not in vivo

MUC1 antigen is recognized as a high‐molecular‐weight glycoprotein that is unexpectedly over‐expressed in human breast and other carcinomas. In contrast, C595 a monoclonal antibody (mAb) against the protein core of the human urinary epithelial machine, is commonly expressed in breast carcinomas. The aim of this study was to conjugate ultra‐small super paramagnetic iron oxide nanoparticles (USPIO) with C595 mAb, in order to detect in vivo MUC1 expression. A dual contrast agent (the C595 antibody‐conjugated USPIO labeled with 99mTc) was prepared for targeted imaging and therapy of anti‐MUC1‐expressing cancers. The C595 antibody‐conjugated USPIO had good stability and reactivity in the presence of blood plasma at 37 °C. No significant differences were observed in immunoreactivity results between conjugated and nonconjugated nanoparticles. The T₁ and T₂ measurements show >79 and 29% increments (for 0.02 mg/ml iron concentrations) in T₁ and T₂ values for USPIO‐C595 in comparison with USPIO, respectively. The nanoprobes showed the interesting targeting capability of finding the MUC1‐positive cell line in vitro. However, we found disappointing in vivo results (i.e. very low accumulation of nanoprobes in the targeted site while >80% of the injected dose per gram was taken up by the liver and spleen), not only due to the coverage of targeting site by protein corona but also because of absorption of opsonin‐based proteins at the surface of nanoprobes. Copyright © 2014 John Wiley & Sons, Ltd.     

Color Doppler ultrasound for evaluation of vasomotor activity in patients with carpal tunnel syndrome

Background  

Patients with carpal tunnel syndrome (CTS) have a variety of vasomotor symptoms. Here, we aimed to study the vasomotor activity of the radialis indicis (RI) artery (median nerve territory) and the radial palmar digital (RPD) artery of the little finger (ulnar nerve territory) before and after sympathetic stimulation in CTS patients using color Doppler ultrasound.