Impact of regulatory T cell therapy on immune cell composition and fetal survival rate in abortion prone mice.

CONTEXT: Implantation of fertilised eggs and survival of a semi-allogenic embryo rely on the interactions between the cells and molecules preparing the uterus. We investigated the effect of regulatory T cell (Treg) therapy on the mechanism of local immune tolerance of mice prone to spontaneous abortion. METHODS: Naive T cells were stimulated in vitro with 17ß-oestradiol (E2), progesterone (P4) and TGF-ß1 for 96h to generate induced Tregs (iTreg). The iTregs were injected into DBA/2-mated pregnant CBA/J female mice (abortion prone model). On day 14 of pregnancy, mice were killed and decidual and placental tissues were collected for cellular composition analysis. RESULTS: Abortion prone mice (PBS treated) showed significantly lower survival rates (P <0.0001), increased CD3+ CD8+ (P <0.05), lower IDO+ (P <0.05) and increased natural killer cells (uNK) cell numbers (P <0.001) in the uterus, as well increased NK cells in the placenta (P <0.05) than in normal pregnant mice (CBA/J×BALB/c). Adoptive transfer of iTregs increased fetal survival in abortion-prone mice (P <0.01) and histopathological evaluation revealed a significantly decreased number of uNK cells in the uterus of TGF-ß1-, E2- and P4-iTregs (P<0.05, P<0.0001 and P<0.05, respectively) than in the PBS treated group. In the placenta, we found significantly lower numbers of uNK cells from TGF-ß1-, E2- and P4-iTregs than in the PBS treated group (P <0.05, P <0.05 and P <0.01, respectively). CONCLUSIONS: We propose that modulation of uterine NK cell activity through immunotherapy using Treg cells should be given more attention as an immunological strategy in the treatment of recurrent miscarriage.

Microbial Profile and Antibiotic Susceptibility Pattern in Diabetic Patients with Mild, Moderate, and Severe Foot Infections in Tehran.

It is estimated that 10-25% of diabetic patients will encounter diabetic foot ulcers (DFU) during their lifetime. This study evaluated the microbiology of DFUs and determined the antibiotic resistance pattern of bacterial isolates based on the severity of wounds and infections in different grades of ulcer. The specimens were collected from115 diabetic foot infections (DFI) deep tissue by needle aspiration and biopsy. The aerobic and anaerobic cultures and antimicrobial susceptibility testing were carried out. The presence of resistance genes including metallo-beta-lactamases (MBL), extended-spectrum ß-lactamase (ESBL), ermA, ermC, and mecA was also determined. A total of 222 microorganisms were isolated. The prevalence of poly-microbial infections was 69.6%. Bacterial isolates comprised 64.2% Gram-positive bacteria (GPB), 33.5% Gram-negative bacteria (GNB), and five isolates of anaerobic bacteria were also detected. The most prevalent GPB and GNB were Staphylococcus spp. (52.2%) and Escherichia coli (33.3%), respectively. The prevalence of poly-microbial infections and GNB was positively associated with increased grades of Wagner and IDSA classifications. Among Staphylococcus aureus isolates, resistance to clindamycin (73.5%), ciprofloxacin (70.6%), and erythromycin (70.6%) were noticeable. GNB was also highly resistant to cephalosporins and ciprofloxacin. ESBL genes were detected in approximately 40% of isolates of Enterobacteriaceae. The prevalence of ermA, ermC, and mecA genes in S. aureus isolates were 8.8%, 32.3%, and 14.7%, respectively. In conclusion, our data suggest that GPBs are the most common isolates from DFIs. Furthermore, with the development of wounds and infection, the prevalence of GNB in DFIs are increased.

Anisotropic damage of soft tissues in supra-physiological deformations.

Soft tissues may undergo mechanical damage under supra-physiological deformations caused by medical treatments such as balloon-angioplasty and stent deployment. This damage is exhibited as a softening in the mechanical behavior of tissues. In this work, alteration of the collagen network is treated as the origin of damage and loss of stiffness. Inspired by the hierarchical structure of the collagen network, the mechanical properties of collagenous tissues are connected to model parameters. Softening of esophageal and arterial tissues under directional cyclic loading is investigated to determine evolution of the associated material parameters through damage progress. An evolution law is proposed which predicts the mechanical behavior of tissues after excessive deformations. Various deformation measures are examined as the damage parameter to determine the most appropriate one for general 3D loading. It is observed that, if the Green-Lagrange strain in the direction of the fibers is used as the damage parameter, the proposed law well describes the evolution of the collagen network's stiffness. The results not only facilitate prediction of the deformation-induced damage under supra-physiological deformations but also are useful for surgeons in better planning of surgical procedures and stents design.

Adoptive cell therapy with induced regulatory T cells normalises the abortion rate in abortion-prone mice.

Ovarian hormones drive invivo generation of regulatory T cells (Tregs) during pregnancy. Little is known about the therapeutic potential of invitro hormone-derived Tregs in pregnancy loss. We investigated the effects of hormone-induced Tregs in a murine model of abortion. CD4+CD25- T cells were isolated from the spleens of CBA/J mice and stimulated with either 17ß-oestradiol (E2), progesterone (P4) or transforming growth factor-ß1 (TGFB1) plus retinoic acid (RA) for 4 days to generate induced Tregs (iTregs). On Days 1-4 of gestation, DBA/2-mated pregnant CBA/J female mice (abortion prone) were injected intravenously with iTregs or Tregs isolated from normal BALB/c-mated pregnant CBA/J mice (np-Tregs). On Day 14, the number of resorbed fetuses was assessed. Serum interferon (IFN)-γ and uterine forkhead box p3 (Foxp3) expression was analysed by ELISA and immunohistochemistry respectively. Using a 3H-thymidine incorporation assay, isolated CD4+CD25+ Tregs induced by the different treatments suppressed the proliferation of CD4+CD25- T cells. Adoptive transfer of iTregs (from all induction groups) significantly decreased fetal resorption in abortion-prone mice. There were no significant changes in serum IFN-γ concentrations after the adoptive transfer of iTregs or np-Tregs. Immunohistochemistry revealed significantly higher Foxp3 expression in gravid uteri from mice injected with np-Tregs and P4-induced iTregs than in the phosphate-buffered saline-treated group. The findings of this study indicate a potential therapeutic benefit of invitro-induced Tregs in patients with recurrent abortion.

Comparison home care service versus hospital-based care in patients with diabetic foot ulcer: an economic evaluation study.

INTRODUCTION: Providing health care to patients at home could be causing the mortality and readmission rates reduction in addition to satisfaction of both patients and health care providers increase. The aim of this study was to assess the cost-effectiveness of home care service compared to hospital based care in patients with diabetic foot ulcer. METHODS: An economic evaluation study and a trial study were simultaneously conducted in Iran. In trial phase, patients with diabetic foot ulcer were randomly assigned to the home care or hospital care. The Cost and Quality of life data were determined as measures of the study. Incremental cost-effectiveness ratio was calculated for comparative purposes. The model consisted of five stages of the disease. The Tree Age Pro 2009 and R software's were used for data analysis. RESULTS: 120 patients were enrolled in our trial; among which 30 patients were in home care service group and 90 patients in hospital based care group. The rate of ulcer size reduction in hospital based care was significant (P value = 0.003) in comparison with home care service. The total cost of the home care and hospital strategies were 1720.4 US$, 3940.3 US$ and the total effectiveness were 0.31 and 0.29, respectively. The incremental cost-effectiveness ratio (ICER) was 117,300 US$ per quality-adjusted life year for home care intervention compared to hospital based care. Based on ICER plane home care treatment will be placed on the southeastern quadrant of the Cost-Effectiveness Plane, and is suggested as a more dominant treatment alternative. CONCLUSIONS: Regarding current evidence, home care strategy for patients suffering diabetic foot ulcer enjoys more cost effectiveness compared to hospital care. It is suggested that healthcare policy makers determine the tariff for health care services for disease groups according to the activity based costing approach.

Spectrofluorimetric method for determination of bupropion in pharmaceutical dosage forms.

OBJECTIVE: Bupropion is a noradrenaline and dopamine reuptake inhibitor which is used as an antidepressant drug. Few HPLC and spectrophotometric methods have been reported before for the determination of bupropion. Most of the previous methods reported for determination of bupropion in pharmaceutical dosage forms are somehow dangerous to health and environment because of using organic solvents. METHOD: In the present method bupropion was determined in pharmaceutical dosage forms by spectrofluorimetry after ion-pair complex formation with eosin Y. The ion-pair complex formation was optimized for reagent amount, buffer pH and time. RESULT: The developed method was linear over the range of 3-120µgmL-1 with an acceptable precision (CV<1.5%) and accuracy (Error<1%). CONCLUSION: The present method is applicable for determination of bupropion in pharmaceutical dosage forms for routine quality control analysis.

Expression profiling of plac1 in murine cancer cell lines

AIM: Placenta-specific 1 (PLAC1) is among recently-discovered placental antigens which exerts fundamental role in placental function and development. Increasing body of literature shows that PLAC1 is frequently activated and expressed in a wide variety of human cancers and promote cancer progression. However, no data is available regarding the expression of mouse orthologue, plac1, in murine cancer cell lines. Materials and Methods: We investigated the expression of plac1 in a series of murine cell lines from different histological origins, mammary carcinoma (4T1), melanoma (B16F10), colorectal carcinoma (CT26), renal carcinoma (Renca), glioma (GL26), B-cell lymphoma (A20 and BCL1) and also two fibroblast cell lines (NIH3T3 and L929), using RT-PCR, Western blotting and flow cytometry. Results: Our data demonstrated that plac1 transcript and plac1 protein were expressed in all examined cell lines, as judged by RT-PCR and Western blot, respectively. The molecular weight of mouse plac1 was experimentally observed to be approximately 24 kD. Flow cytometric analysis showed surface expression of plac1 in aforesaid cell lines ranging from 2% to 42.5%. Conclusion: Based on the ubiquitous expression of plac1, the investigated cancer cell lines or immortalized cell lines can be used to examine the role of plac1 in the process of immortalization.

Evaluation of the immune response in BALB/c mice induced by a novel DNA vaccine expressing GRA14 against Toxoplasma gondii.

Toxoplasma gondii can cause severe and even fatal disease in human beings and animals. Effective vaccines may contribute to control toxoplasmosis. GRA14, a novel secreted dense granule protein of T. gondii, has been proposed as a vaccine candidate due to its intervacuolar transport and unique topology in the parasitophorous vacuole membrane. In this study, we constructed a DNA vaccine encoding GRA14 of T. gondii. BALB/c mice were immunized intramuscularly three times at 2 week intervals and challenged with T. gondii RH strain 5 weeks later. The immune responses were evaluated using lymphocyte proliferation assay, cytokine and antibody measurements. In addition, the survival times and parasite load of mice challenged with the virulent T. gondii RH strain were evaluated. The results showed that the mice immunized with pcGRA14 induced both enhanced specific humoral and Th1 cellular immune responses, and also mice immunized with the pcGRA14 showed an increased survival time and decreased parasite load compared with control groups (P<.05). The results indicated, for the first time, that the GRA14 is a potential DNA vaccine against toxoplasmosis.

The metabolomics and lipidomics window into thyroid cancer research.

CONTEXT: Thyroid carcinoma is the most common endocrine system malignancy with a fast rising incidence in the last decade for unknown reasons. Fine needle aspiration (FNA) biopsy, the gold standard in thyroid cancer (TC) screening has still its own challenges and in some cases needs a proceeding surgery. OBJECTIVE: This review highlights the role of the two most recent "omics" approaches, "metabolomics" and "lipidomics", in the field of TC research. METHODS: All the previous studies have been extracted from the literature and major concepts were detailed in the field of TC metabolomics and lipidomics. RESULTS: Metabolomics and lipidomics, have potential in finding biomarkers related to thyroid carcinoma. Among the previous studies, the most important introduced altered tissue metabolites and lipids included glucose and galactose, lactate, Scyllo- and Myo inositol, hypoxanthine, citrate, cholesterol and choline. CONCLUSION: Metabolomics methods have been widely used in the field of biomarker discovery in TC and attempts are still in progress to use these methods to find a reliable biomarker panel besides current diagnostic tools.

Formulation, in vitro evaluation and kinetic analysis of chitosan-gelatin bilayer muco-adhesive buccal patches of insulin nanoparticles.

The present study was performed to optimise the formulation of a muco-adhesive buccal patch for insulin nanoparticles (NPs) delivery. Insulin NPs were synthesised by an ionic gelation technique using N-di methyl ethyl chitosan cysteine (DMEC-Cys) as permeation enhancer biopolymer, tripolyphosphate (TPP) and insulin. Buccal patches were developed by solvent-casting technique using chitosan and gelatine as muco-adhesive polymers. Optimised patches were embedded with 3 mg of insulin-loaded NPs with a homogeneous distribution of NPs in the muco-adhesive matrix, which displayed adequate physico-mechanical properties. The drug release characteristics, release mechanism and kinetics were investigated. Data fitting to Peppas equation with a correlation coefficient indicated that the mechanism of drug release followed an anomalous transport that means drug release was afforded through drug diffusion along with polymer erosion. In vitro drug release, release kinetics, physical and mechanical studies for all patch formulations reflected the ideal characteristics of this buccal patch for the delivery of insulin NPs.

Effect of smoking on retrobulbar blood flow in thyroid eye disease.

PurposeTo evaluate the effect of smoking on retrobulbar blood flow parameters by color Doppler imaging in patients with thyroid eye disease.Patients and methodsIn this observational case series, blood flow parameters in the ophthalmic artery, superior ophthalmic vein, central retinal artery, and vein were determined by color Doppler imaging in patients with thyroid eye disease. Patients were grouped as smokers and non-smokers. Never smokers and those who had stopped smoking for at least 1 year before onset of ophthalmopathy were considered as non-smokers. A thorough ophthalmic examination including Hertel exophthalmometry was performed. Orbital CT scan was also carried out in all patients.ResultsFifty-one orbits from 30 patients between 21 and 62 years old (mean±SD: 40.8±12.0) were enrolled in this study. Smokers had greater proptosis and more active and sever disease. (P<0.05) Muscle involvement based on CT scan did not vary in smokers and non-smokers. Maximum velocity (3.78±1.74 vs 5.27±2.14, P<0.001; t-test) and minimum velocity (1.74±0.78 vs 3.26±1.36, P =0.014; t-test) in superior ophthalmic vein were significantly lower in smokers than non-smokers. Even after adjusting for age, sex, and clinical activity score and severity, smokers had a lower minimum velocity in superior ophthalmic vein (P =0.01; ANCOVA analysis).ConclusionCigarette smoking may correlate with increase in orbital venous congestion in thyroid eye disease.

Effect of socks structures on plantar dynamic pressure distribution.

A major purpose of investigating the plantar pressure in patients with pain or those at risk for skin injury is to reduce the pressure below metatarsal heads, specially first and second metatarsal heads. The aim of this article is to evaluate the effects of the socks structures on the changes in plantar dynamic pressure. In this study, seven socks types with different structures for the sole area were produced. The Gaitview® AFA-50 system, a force plate, was used to measure the plantar dynamic pressure of 10 participants. The barefoot plantar dynamic pressure distribution was compared with the plantar dynamic pressure distribution with socks by two independent samples test on various zones of the foot and on different genders using SPSS software. Mann-Whitney tests were used to determine specific significant differences. The obtained results showed that the main trend was to redistribute the plantar dynamic pressure from the higher plantar pressure zones (toe and first through forth metatarsal bone regions) were decreased and as a result the plantar pressure toward the relatively lower pressure zones (fifth metatarsal bone and midfoot regions). In comparison with the barefoot condition, the cross miss structure reduced the mean pressure in the critical region of the foot (first metatarsal) for male and female subjects ( p < 0.05) and also the mock rib structure reduced the mean pressure for female subjects ( p < 0.05). In general, the results suggested wearing the socks because the socks make the plantar pressure redistributed from high to low plantar pressure zones. The results of this research indicated that wearing socks with cross miss and mock rib structures will reduce the mean plantar pressure values in forefoot area in comparison with the barefoot condition.

Nonlinear damping and quasi-linear modelling.

The mechanism of energy dissipation in mechanical systems is often nonlinear. Even though there may be other forms of nonlinearity in the dynamics, nonlinear damping is the dominant source of nonlinearity in a number of practical systems. The analysis of such systems is simplified by the fact that they show no jump or bifurcation behaviour, and indeed can often be well represented by an equivalent linear system, whose damping parameters depend on the form and amplitude of the excitation, in a 'quasi-linear' model. The diverse sources of nonlinear damping are first reviewed in this paper, before some example systems are analysed, initially for sinusoidal and then for random excitation. For simplicity, it is assumed that the system is stable and that the nonlinear damping force depends on the nth power of the velocity. For sinusoidal excitation, it is shown that the response is often also almost sinusoidal, and methods for calculating the amplitude are described based on the harmonic balance method, which is closely related to the describing function method used in control engineering. For random excitation, several methods of analysis are shown to be equivalent. In general, iterative methods need to be used to calculate the equivalent linear damper, since its value depends on the system's response, which itself depends on the value of the equivalent linear damper. The power dissipation of the equivalent linear damper, for both sinusoidal and random cases, matches that dissipated by the nonlinear damper, providing both a firm theoretical basis for this modelling approach and clear physical insight. Finally, practical examples of nonlinear damping are discussed: in microspeakers, vibration isolation, energy harvesting and the mechanical response of the cochlea.

Does endurance training affect IGF-1/IGFBP-3 and insulin sensitivity in patients with type 2 diabetes?

AIM: The aim of the present study was to determine whether six weeks of submaximal endurance training using a cycle ergometer would result in a modified serum insulin-like growth factor-1 (IGF-1), an insulin-like growth factor binding protein 3 (IGFBP-3), and insulin resistance in middle-aged men with type 2 diabetes (T2D). METHODS: Twenty male patients with T2D voluntarily participated in this study and were randomly divided into two groups: the training group (N.=10) and the control group (N.=10). The training protocol consisted of a 45-minutes cycling session/day, three days/week for six weeks with intensity 60-70% of the maximum heart rate. To examine the IGF-1 and the IGFBP-3, fasting blood glucose levels, and insulin resistance, blood sampling was performed before and immediately after the first and 18th sessions. The homeostatic model assessment (HOMA-IR) method was used to determine insulin resistance. RESULTS: Before the study began, no significant difference between the two groups was observed in the anthropometric and blood factors. After a session of aerobic exercise, IGF-1 and IGFBP-3 levels were significantly increased (153.79% and 64.3%, respectively), and fasting glucose and insulin resistance levels were significantly decreased (15.82% and 27.82%, respectively); however, the changes resulting from a six-week training period were not significant. CONCLUSION: According to the present study, one session of aerobic exercise for middle-aged men with T2D leads to increased IGF-1 and IGFBP-3, and to decreased fasting glucose and insulin resistance. Considering the lack of changes after a six-week training, it seems that the amount of change depends on subjects' fitness level and exercise parameters. From a clinical point of view, the beneficial effects of acute exercise inT2D subjects show that such exercises should be part of the daily program for them.

Reusable and robust high sensitive non-enzymatic glucose sensor based on Ni(OH)2 nanoparticles.

Nickel hydroxide nanoparticles were successfully electrodeposited on graphite electrode (Gr/NiONP) and employed as a robust non-enzymatic glucose sensor. The results of cyclic voltammetry (CV) and chronoamperometry demonstrated that the Gr/NiONP electrode displayed high electrocatalytic activity toward glucose. The oxidation current is directly related to the glucose concentration from 1 µM to 15 mM. Besides, the glucose sensor displayed high sensitivity (2400 µA mM(-1) cm(-2)) with a detection limit of 0.53 µM (S/N=3) in basic solution. Moreover, the sensor showed excellent selectivity, reproducibility and stability properties. The relative standard deviation is 1.2% for 10 successive measurements in 16 µM glucose. Interestingly, the signal for glucose was maintained at 95% of its initial value even after 6 months of storage under ambient conditions. Gr/NiONP electrode has also been tested to detect glucose in human serum with satisfactory results.

Polyphyly, gene-duplication and extensive allopolyploidy framed the evolution of the ephemeral Vulpia grasses and other fine-leaved Loliinae (Poaceae).

The fine-leaved Loliinae is one of the temperate grass lineages that is richest in number of evolutionary switches from perennial to annual life-cycle, and also shows one of the most complex reticulate patterns involving distinct diploid and allopolyploid lineages. Eight distinct annual lineages, that have traditionally been placed in the genus Vulpia and in other fine-leaved ephemeral genera, have apparently emerged from different perennial Festuca ancestors. The phenotypically similar Vulpia taxa have been reconstructed as polyphyletic, with polyploid lineages showing unclear relationships to their purported diploid relatives. Interspecific and intergeneric hybridization is, however, rampant across different lineages. An evolutionary analysis based on cloned nuclear low-copy GBSSI (Granule-Bound Starch Synthase I) and multicopy ITS (Internal Transcribed Spacer) sequences has been conducted on representatives of most Vulpia species and other fine-leaved lineages, using Bayesian consensus and agreement trees, networking split graphs and species tree-based approaches, to disentangle their phylogenetic relationships and to identify the parental genome donors of the allopolyploids. Both data sets were able to reconstruct a congruent phylogeny in which Vulpia was resolved as polyphyletic from at least three main ancestral diploid lineages. These, in turn, participated in the origin of the derived allopolyploid Vulpia lineages together with other Festuca-like, Psilurus-like and some unknown genome donors. Long-distance dispersal events were inferred to explain the polytopic origin of the Mediterranean and American Vulpia lineages.

Gram-negative bacteremia: which empirical antibiotic therapy?

PURPOSE: Given the increasing frequency of cefotaxime-resistant strains, third-generation cephalosporins (3GC e.g. cefotaxime, ceftriaxone) might not be recommended any longer as empirical antibiotic therapy for community-acquired Gram-negative bacteremia (CA-GNB). PATIENTS AND METHODS: We conducted a multicenter prospective descriptive study including patients with CA-GNB. RESULTS: Two hundred and nineteen patients were included. Escherichia coli and Pseudomonas aeruginosa were the most frequently isolated species in 63% (n=138) and 11% (n=24) of the cases, respectively. The prevalence of cefotaxime-resistance reached 18% (n=39) mostly due to intrinsic resistance (27 cases, 12%). The presence of invasive material (P<0.001), the origin of the patient (Paris region or West of France) (P=0.006), and home health care (P<0.001) were variables predicting resistant GNB. The negative predictive value for resistance in patients with invasive material coming from the West of France, or without invasive material and with home health care was 94%. The positive predictive value for patients with invasive material living in Paris, or without invasive material and with home health care only reached 58 and 54%, respectively. CONCLUSIONS: Using 3GC for CA-GNB due to cefotaxime-resistant strains was relatively frequent, ESBL-producing Enterobacteriaceae being rarely involved. Our study highlights the role of local epidemiology; before any changes to first-line antibiotic therapy, local epidemiological data should be taken into account.

Application of artificial neural networks for optimization of preparation of insulin nanoparticles composed of quaternized aromatic derivatives of chitosan.

The aim of this research was to develop an artificial neural network (ANN) in order to design a nanoparticulate oral drug delivery system for insulin. The pH of polymer solution (X1), concentration ratio of polymer/insulin (X2) and polymer type (X3) in 3 level including methylated N-(4-N,N- dimethyl aminobenzyl) chitosan, methylated N-(4-pyridinyl) chitosan, and methylated N-(benzyl) chitosan are considered as the input values and the particle size, zeta potential, PdI, and entrapment efficiency (EE %) as output data. ANNs are employed to generate the best model to determining the relationships between input and response values. In this research, a multi-layer percepteron with different topologies has been tested in order to define the one with the best accuracy and performance. The optimization was used by minimizing the error between the predicted and observed values. Three training algorithms (Levenberg-Marquardt (LM), Bayesian-Regularization (BR), and Gradient Descent (GD)) were employed to train ANNs with various numbers of nodes, hidden layers and transfer functions by random selection. The accuracy of prediction data were assayed by the mean squared error (MSE).The ability of all algorithms was in the order: BR>LM>GD. Thus, BR was selected as the best algorithm.