<Emphasis Type="Italic">Terminalia bellirica</Emphasis> fruit extracts: in-vitro antibacterial activity against selected multidrug-resistant bacteria,radical scavenging activity and cytotoxicity study on BHK-21 cells

Background

Identification of novel sources for developing new antibiotics is imperative with the emergence of antibiotic resistant bacteria. The fruits of Terminalia bellirica (Gaertn) Roxb., widely used in traditional medicine, were evaluated for antibacterial activity against multidrug-resistant (MDR) bacteria, antioxidant activity and cytotoxicity.

Methods

Twelve solvent extracts of T. bellirica fruits were prepared by direct aqueous extraction and sequential extraction with dichloromethane, methanol and water using Soxhlet, bottle-shaker and ultrasound sonicator methods. Antibacterial activity of the extracts was tested against 16 strains MDR bacteria—methicillin-resistant Staphylococcus aureus (MRSA), extended spectrum β-lactamase (ESBL) producing Escherichia coli and MDR Acinetobacter spp., Klebsiella pneumoniae and Pseudomonas aeruginosa—and 4 control organisms, using the cut-well diffusion method. The minimum inhibitory concentration (MIC) was determined using an agar dilution method. The radical scavenging activity of six antibacterial extracts was screened against 2,2′-diphenyl-2-picrylhydrazyl (DPPH) and correlation was established between EC₅₀ (50% effective concentration) values and the total phenolic content (TPC). Cytotoxicity was determined for the most potent antibacterial extract on baby hamster kidney (BHK-21) cells by Tryphan Blue exclusion method. Statistical analysis was carried out by one-way analysis of variance at significant level p?<?0.05 using “SigmaPlot 10” and “R 3.2.0” software.

Results

All aqueous and methanol extracts displayed antibacterial activity (MIC 0.25–4?mg/mL) against all strains of MRSA, MDR Acinetobacter spp. and MDR P. aeruginosa. The sequential aqueous extracts (MIC, 4?mg/mL) inhibited ESBL producing-E. coli. None of the extracts exhibited activity against MDR K. pneumoniae (MIC >?5?mg/mL). The sequential methanol extract (Soxhlet) recorded high antibacterial activity and the highest DPPH radical scavenging activity (EC₅₀, 6.99?±?0.15?ppm) and TPC content (188.71?±?2.12 GAE mg/g).The IC₅₀ (50% inhibition concentration) values of the most potent antibacterial extract—the direct aqueous extract from reflux method—on BHK-21 cells were 2.62?±?0.06 and 1.45?±?0.08?mg/ml with 24 and 48?h exposure, respectively.

Conclusions

Results indicate that T. bellirica fruit is a potential source for developing broad-spectrum antibacterial drugs against MDR bacteria, which are non-toxic to mammalian cells and impart health benefits by high antioxidant activity.

     

Asthma and Hypogammaglobulinemia: an Asthma Phenotype with Low Type 2 Inflammation

Purpose

Little is known about hypogammaglobulinemia (HGG) in asthma patients. No data are available on the characteristics of adult patients with asthma and HGG.

Methods

We conducted a retrospective monocentric study between January 2006 and December 2012. Asthma patients with a serum immunoglobulin (Ig) quantitative analysis were included and classified into two groups depending on their serum IgG concentration: presence or absence of HGG. Clinical, biological, functional, and radiologic characteristics were compared in univariate and multivariate analysis, using a logistic regression model.

Results

In univariate analysis, asthma patients with HGG (n?=?25) were older (58 years old?±?18 vs 49?±?18, p?=?0.04) and more frequently active or former smokers as compared to patients with normoglobulinemia (n?=?80) (56.0 vs 35.0 %, p?=?0.01). Total IgE?<?30 kUI/L was more frequently observed in patients with HGG (53.0 vs 18.3 %, p?=?0.01). HGG asthma patients had lower fraction of exhaled nitric oxide (p?=?0.02), blood eosinophilia (p?=?0.0009), and presented with more severe composite score for bronchiectasis (p?=?0.01). In multivariate analysis, asthma patients with HGG had increased risk of being smokers [OR?=?6.11 (IC 95 %?=?1.16–32.04)], having total IgE concentration?<?30 kUI/L [OR?=?12.87 (IC 95 %?=?2.30–72.15)], and a more severe composite score of bronchiectasis [OR?=?20.65 (IC 95 %?=?2.13–199.74)].

Conclusion

Asthma patients with HGG are older and more often tobacco smoker than asthma patients without HGG. These patients have low type-2 inflammation markers.

     

Therapeutic properties in Tunisian hot springs: first evidence of phenolic compounds in the cyanobacterium <Emphasis Type="Italic">Leptolyngbya</Emphasis> sp. biomass,capsular polysaccharides and releasing polysaccharides

Background

In Tunisia, the use of hot spring waters for both health and recreation is a tradition dating back to Roman times. In fact, thermal baths, usually called “Hammam” are recommended as a therapeutic and prophylactic measure against many types of illness and toxicity. While the chemical concentration of thermal water is admittedly associated with its therapeutic effects, the inclusion in spa waters of efficient bioproduct additives produced by photosynthetic microorganisms and that act against oxidative stress may comprise a significant supplementary value for thermal centers. The aim of this study was to investigate the antioxidant potential of the Tunisian thermophilic cyanobacterium Leptolyngbya sp. and to determine its phytochemical constituents and phenolic profile.

Methods

BME (Biomass Methanolic Extract), CME (Capsular polysaccharides Methanolic Extract) and RME (Releasing polysaccharides Methanolic Extract) of Leptolyngbya sp. were examined for their antioxidant activities by means of DPPH, hydroxyl radical scavenging and ferrous ion chelating assays. Their total phenols, flavonoids, carotenoids, Mycosporine-like amino acids (MAAs) and vitamin C contents, as well as their phenolic profiles were also determined.

Results

BME has the highest content of phenols (139?±?1.2 mg/g), flavonoids (34.9?±?0.32 mg CEQ/g), carotenoids (2.03?±?0.56 mg/g) and vitamin C (15.7?±?1.55 mg/g), while the highest MAAs content (0.42?±?0.03 mg/g) was observed in CME. BME presented both the highest DPPH and hydroxyl radical scavenging ability with an IC₅₀ of 0.07 and 0.38 mg/ml, respectively. The highest ferrous chelating capacity was detected in CME with an IC₅₀?=?0.59 mg/ml. Phenolic profiles revealed the presence of 25 phenolic compounds with the existence of hydroxytyrosol, oleuropein, resveratrol and pinoresinol.

Conclusion

The study demonstrated that the cyanobacterium Leptolyngbya sp. possesses abundant natural antioxidant products which may have prophylactic and therapeutic effects on many types of illness and toxicity. The present findings not only explain and reinforce the rationale behind traditional therapeutic practices in Tunisia in the exploitation of the country’s hot springs, but support the addition of Leptolyngbya to thermal waters as a means to enhance the value and reputation of the curative nature of Tunisian thermal waters.

     

Modification Effects of Changes in Job Demands on Associations Between Changes in Testosterone Levels and Andropause Symptoms: 2-Year Follow-up Study in Male Middle-Aged Japanese Workers

Purpose

The purpose of this longitudinal study was to ascertain if changes in job demands modify associations between changes in testosterone levels and andropause symptoms in male Japanese workers.

Method

A baseline survey including job demands and the Aging Males’ Symptoms scale, lifestyle factors, and blood levels of testosterone was conducted in 2007. Among 192 men (mean age?±?SD 52.2?±?7.6 years) who completed all relevant questionnaires and provided blood at baseline, 104 men (50.9?±?7.2 years) were followed up in 2009. Changes of variables in 2 years were calculated (data of follow-up minus those of baseline).

Results

Testosterone levels were increased significantly, whereas job demands and somatic symptoms were reduced significantly, at follow-up. Changes in testosterone levels were negatively associated with changes in total andropause symptoms, psychological symptoms, and sexual symptoms (standardized β?=??0.27, ?0.24, and, ?0.29, p?<?0.05, respectively), after adjustment for confounders. Changes in job demands were positively associated with changes in somatic symptoms (standardized β?=?0.21, p?<?0.05). Significant interactions of changes in testosterone levels and job demands were noted for changes in psychological symptoms (standardized β?=?0.26, p?<?0.05). For men with a 1-SD reduction in job demands, negative associations between changes in testosterone levels and psychological symptoms were intensified, but not for men with a 1-SD increase in job demands.

Conclusion

Andropause symptoms may be affected by changes in testosterone levels and job demands. Change in job demands may modify associations between changes in testosterone levels and andropause symptoms.

     

<Emphasis Type="Italic">Helicobacter bilis</Emphasis>-Associated Suppurative Cholangitis in a Patient with X-Linked Agammaglobulinemia

?

Helicobacter bilis is a commensal bacterium causing chronic hepatitis and colitis in mice. In humans, enterohepatic Helicobacter spp. are associated with chronic hepatobiliary diseases.

Purpose

We aimed at understanding the microbial etiology in a patient with X-linked agammaglobulinemia presenting with suppurative cholangitis.

Methods

16S rDNA PCR directly performed on a liver biopsy retrieved DNA of H. bilis.

Results

Clinical outcome resulted in the normalization of clinical and biological parameters under antibiotic treatment by a combination of ceftriaxone, metronidazole, and doxycyclin followed by a 2-week treatment with moxifloxacin and a 2-month treatment with azithromycin.

Conclusion

In conclusion, these data suggest a specific clinical and microbiological approach in patients with humoral deficiency in order to detect H. bilis hepatobiliary diseases.

     

Development of an algorithm to automatically compress a CT image to visually lossless threshold

Background

To develop an algorithm to predict the visually lossless thresholds (VLTs) of CT images solely using the original images by exploiting the image features and DICOM header information for JPEG2000 compression and to evaluate the algorithm in comparison with pre-existing image fidelity metrics.

Methods

Five radiologists independently determined the VLT for 206 body CT images for JPEG2000 compression using QUEST procedure. The images were divided into training (n?=?103) and testing (n?=?103) sets. Using the training set, a multiple linear regression (MLR) model was constructed regarding the image features and DICOM header information as independent variables and regarding the VLTs determined with median value of the radiologists’ responses (VLT_rad) as dependent variable, after determining an optimal subset of independent variables by backward stepwise selection in a cross-validation scheme.The performance was evaluated on the testing set by measuring absolute differences and intra-class correlation (ICC) coefficient between the VLT_rad and the VLTs predicted by the model (VLT_model). The performance of the model was also compared two metrics, peak signal-to-noise ratio (PSNR) and high-dynamic range visual difference predictor (HDRVDP). The time for computing VLTs between MLR model, PSNR, and HDRVDP were compared using the repeated ANOVA with a post-hoc analysis. P?<?0.05 was considered to indicate a statistically significant difference.

Results

The means of absolute differences with the VLT_rad were 0.58 (95% CI, 0.48, 0.67), 0.73 (0.61, 0.85), and 0.68 (0.58, 0.79), for the MLR model, PSNR, and HDRVDP, respectively, showing significant difference between them (p?<?0.01). The ICC coefficients of MLR model, PSNR, and HDRVDP were 0.88 (95% CI, 0.81, 0.95), 0.85 (0.79, 0.91), and 0.84 (0.77, 0.91). The computing times for calculating VLT per image were 1.5?±?0.1 s, 3.9?±?0.3 s, and 68.2?±?1.4 s, for MLR metric, PSNR, and HDRVDP, respectively.

Conclusions

The proposed MLR model directly predicting the VLT of a given CT image showed competitive performance to those of image fidelity metrics with less computational expenses. The model would be promising to be used for adaptive compression of CT images.

     

The relationship between lymphovascular invasion and angiogenesis,hormone receptors,cell proliferation and survival in patients with primary operable invasive ductal breast cancer

Background

Several well-established tumour prognostic factors are used to guide the clinical management of patients with breast cancer. Lymphovascular invasion and angiogenesis have also been reported to have some promise as prognostic factors. The aim of the present study was to examine the prognostic value of tumour lymphovascular invasion and microvessel density compared with that of established prognostic factors in invasive ductal breast cancer.

Methods

In addition to hormone receptor status and Ki-67 proliferative activity, lymphovascular invasion and microvessel density and their relationship with survival were examined in patients with invasive ductal breast cancer. Full sections and tissue microarrays (n?=?384 patients) were utilised to assess these factors and were scored by appropriate methods.

Results

On univariate analysis tumour size (P?<?0.05), lymph node involvement (P?<?0.01), lymphovascular invasion (P?<?0.05), microvessel density (P?<?0.05) and local- regional treatment (P?<?0.01) were associated with poorer survival in ER negative tumours. On multivariate analysis in ER negative tumours lymph node involvement (P?<?0.01) and local- regional treatment (P?<?0.05) were independently associated with poorer cancer-specific survival. On univariate analysis tumour grade (P?<?0.05), lymph node involvement (P?<?0.001), HER-2 (P?<?0.05), Ki-67 (P?<?0.01) and lymphovascular invasion (P?<?0.001) were associated with poorer survival in ER positive tumours. On multivariate analysis lymph node involvement (P?<?0.001), Ki-67 (P?<?0.001) and lymphovascular invasion (P?<?0.05) were independently associated with poorer cancer-specific survival in ER positive tumours.

Conclusion

Lymphovascular invasion but not microvessel density was independently associated with poorer survival in patients with ER positive but not ER negative invasive ductal breast cancer.

     

Ultrasound point shear wave elastography of the pancreas: comparison of patients with type 1 diabetes and healthy volunteers – results from a pilot study

Background

The aims of this study were to establish shear wave elastography of the pancreas by comparing measurements in patients with type 1 diabetes (T1D) and healthy volunteers and to consider whether this method could contribute to the screening or prevention of T1D.

Methods

This pilot study included 15 patients with T1D (10 men, 5 women) and 15 healthy volunteers (10 men, 5 women) as controls. Measurements were performed with a Siemens Acuson S3000 (Siemens Healthcare, Erlangen, Germany) using a 6C1 convex transducer and the Virtual Touch? tissue quantification (VTQ) method.

Results

The mean shear wave velocity of the head of the pancreas was 1.0?±?0.2?m/s (median: 1.1?m/s) for the study group and likewise 1.0?±?0.2?m/s (median: 0.9?m/s) for the control group. Velocities of 1.2?±?0.2?m/s (median: 1.2?m/s) were measured in the body of the pancreas in both groups. There was a significant difference between the values obtained in the tail of the pancreas: patients 1.1?±?0.1?m/s (median: 1.0?m/s) versus controls 0.9?±?0.1?m/s (median: 0.8?m/s) (p?=?0.0474). The mean value in the whole pancreas of the study group was not significantly above that of the control group: 1.1?±?0.1?m/s (median: 1.0?m/s) versus 1.0?±?0.1?m/s (median: 1.0?m/s) (p?=?0.2453).

Conclusions

Sonoelastography of the pancreas revealed no overall difference between patients with T1D and healthy volunteers. Patients with T1D showed higher values only in the tail segment. Future studies need to determine whether specific regional differences can be found in a larger study population.

     

RETRACTED ARTICLE: Optimized extraction,composition, antioxidant and antimicrobial activities of exo and intracellular polysaccharides from submerged culture of <Emphasis Type="Italic">Cordyceps cicadae</Emphasis>

Background

Cordyceps cicadae is known as Jin Chan Hua in Traditional Chinese Medicine and known to possess different pharmacological activities. Presently, it was collected from the wild and isolated. Mycelial culture was optimized for extraction of polysaccharides under submerged culture conditions. Besides antioxidant, antibacterial activities of extracted polysaccharides were tested for first time.

Methods

Exo-polysaccharides (EPS) and intracellular polysaccharides (IPS) production was tested under different factors (medium capacity, rotation speed, pH, incubation time, temperature, carbon, nitrogen, minerals sources and carbon to nitrogen ratio) by orthogonal experiments using one-factor-at-a-time method. Monosaccharides composition of polysaccharides produced by C. cicadae was determined using high performance liquid chromatography. Antioxidant and antimicrobial activities on eight bacterial strains were checked by different standard procedures.

Results

Factors viz., medium capacity, rotation speed, incubation time, pH and temperature affected the EPS and IPS production under submerged culture conditions. EPS and IPS production was observed to vary with different carbon and nitrogen sources as well as C/N ratio. Glucose was the major component of polysaccharides (63.10?±?4.15 %). Extracted EPS and IPS showed higher antioxidant potential with significant DPPH radical scavenging activity, ABTS radical scavenging activity, reducing power and iron chelating activity. Antimicrobial activities of EPS and IPS varied among the tested bacterial strains. IPS showed slightly higher inhibition rate to all the tested bacterial strains as compared to EPS. Maximum inhibition zones of IPS (12.9?±?0.2 mm) and EPS (12.5?±?0.3 mm) was observed against Pseudomonas aeruginosa at 10 % con. However, both EPS and IPS fractions showed broad spectrum for all the pathogenic microbial strains tested. The MIC of both the extracts ranged from 60–100 mg/mL.

Conclusions

EPS and IPS production from submerged culture of C. cicadae with significant antioxidant and antibacterial potential can be enhanced with the combination of several factors which can be used for large scale industrial fermentation of C. cicadae.

     

The association of urinary interferon-gamma inducible protein-10 (IP10/CXCL10) levels with kidney allograft rejection

Background

Interferon-gamma inducible protein-10 (IP-10/CXCL10) is a chemokine involved in the alloimmune response against kidney allograft. We aimed to investigate the association of urinary CXCL10 protein levels with rejection in renal transplant patients.

Methods

A total of 273 urine samples from (biopsy-proven) rejection and non-rejection patients and controls were included in this study. CXCL10 levels were analyzed for association with rejection.

Results

The data showed statistically significant differences in the CXCL10 levels between rejection vs. non-rejection (p?<?0.001). Among the rejection groups, statistically significant differences for CXCL10 levels were found between ACR vs. NAD (p?<?0.001), ACR vs. BLR (p?=?0.019) and AVR vs. NAD (p?=?0.009). Receiver Operating Characteristic (ROC) curve analysis of CXCL10 showed an area under the curve (AUC) of 0.74 with 72% sensitivity and 71% specificity at 27.5 pg/ml between rejection and non-rejection group. Kaplan–Meier curve analysis among different levels of CXCL10 showed a better rejection-free graft survival in patients with <100 pg/ml when compared to >200 pg/ml (38?±?6 vs. 12?±?1.0 weeks; log-rank p?<?0.001) and 100–200 pg/ml (38?±?6 vs. 22?±?9 weeks; log-rank p?=?0.442) concentration.

Conclusion

The results indicate significantly increased levels of CXCL10 protein in the urine at the time of allograft rejection. This association of urinary CXCL10 protein levels with rejection could provide an additional tool for the non-invasive monitoring of allograft rejection.

     

Antidepressant-like effects of methanolic extract of <Emphasis Type="Italic">Bacopa monniera</Emphasis> in mice

Background

Bacopa monniera has been used as a cure for various ailments that include anxiety, epileptic disorders, dementia, blood purifier, cough and rheumatism, and some important local uses of the plant are in dermatitis, anemia, diabetes, promote fertility and prevent miscarriage for many years in Bangladesh. According to this background, the aim of the study was to evaluate the antidepressant-like effect of the methanolic extract of B. monniera (MEBM) in different behavioral models such as forced swimming test (FST), measurement of locomotor activity test (MLAT) and tail suspension test (TST) on mice after two weeks treatment.

Methods

Mice were divided into five groups (n?=?5/group): control group (deionized water), standard group where Imipramine hydrochloride (30 mg/kg) was used as standard drug and three test groups where three doses of the methanolic extract of B. monniera (MEBM) (50, 100, and 200 mg/kg) was used for two weeks treatment. All the drug and test samples were administered via gavage through oral route. To assess the antidepressant-like effect of MEBM forced swimming test (FST), tail suspension test (TST) and measurement of locomotor activity test (MLAT) have been done in mice.

Results

The results showed that a strong and dose-dependent antidepressant effects in different mice models. The main findings of the MEBM significantly reduced the duration of immobility times in the forced swimming test (p?<?0.001). Likewise, the extract significantly decreased the immobility time in the tail suspension test (p?<?0.001). Moreover, we employed an additional measurement of locomotor activity test to check the motor stimulating activity of the MEBM. The extract also significantly increased the locomotion, rearing and defecation effects in comparison to the control group (p?<?0.001).

Conclusion

The present results clearly demonstrate that the methanolic extract of B. monniera possesses antidepressant-like activity in the animal behavioral models. The current study warrants further investigation into identification of the active compounds in herbal medicines, in particular extract of B. monniera with antidepressant-like effects.

     

Evaluation of hepatic markers and body weight gain in growing and finishing steers

Background

The liver plays an important role in nutrient metabolism, in detoxification and excretion of hydrophobic metabolites, and in the synthesis of most circulating proteins. Thus, enhanced knowledge of the processes that regulate liver function in beef cattle production stages can lead to significant improvements in their management.

Aims

The authors studied the levels of some serum liver parameters during growing and finishing stages of steers.

Methods

Blood samples were collected from (n?=?60) steers at day 1 and at day 21 and 42 days after the start of the growing (group 1 n?=?30: mean body weight (BW) 430?±?5.8 Kg) or finishing (group 2 n?=?30: mean BW 591?±?5.3 Kg) phases. Steers from both groups were separated into two subgroups (A and B, respectively) according to the percentage of body weight gain (%WG). Subgroup A steers had a lower mean %WG than subgroup B steers. Serum total proteins (TP), albumin, globulins, serum urea nitrogen (SUN), total and conjugated bilirubin (TB, CB), aspartate amino transferase (AST), and gamma-glutamyltransferase (GGT) were assessed in order to evaluate significant differences between groups and subgroups.

Results

All parameters showed significant differences between the phases and between groups with the exception of AST and GGT. Mean?±?S.D. of %WG was 11.07?±?1.95 for group 1 and 9.84?±?0.77 % for group 2. For group 1 steers, there was a significant difference on SUN concentration and GGT activity between subgroup A and subgroup B.

Conclusions

These findings suggest that hepatic markers in steers are influenced by production stages and improve the knowledge about the effect of growing and finishing phases on their hepatic markers. Furthermore, it underlies the importance of monitoring body weight gain in order to obtain a maximizing profitability.

     

Morphometric study of the two fused primary ossification centers of the clavicle in the human fetus

Purposes

A satisfactory understanding of the clavicle development may be contributing to both the diagnosis of its congenital defects and prevention of perinatal damage to the shoulder girdle. This study was carried out to examine the transverse and sagittal diameters, cross-sectional area and volume of the two fused primary ossification centers of the clavicle.

Methods

Using the methods of CT, digital-image analysis and statistics, the size for two fused primary ossification centers of the clavicle in 42 spontaneously aborted human fetuses at ages of 18–30 weeks was studied.

Results

Without any male–female and right-left significant differences, the best fit growth models for two fused primary ossification centers of the clavicle were as follows: y = ?31.373 + 15.243 × ln(age) ± 1.424 (R ² = 0.74) for transverse diameter, y = ?7.945 + 3.225 × ln(age) ± 0.262 (R ² = 0.78), y = ?4.503 + 2.007 × ln(age) ± 0.218 (R ² = 0.68), and y = ?4.860 + 2.117 × ln(age) ± 0.200 (R ² = 0.73) for sagittal diameters of the lateral, middle and medial ends respectively, y = ?31.390 + 2.432 × age ± 4.599 (R ² = 0.78) for cross-sectional area, and y = 28.161 + 0.00017 × (age)⁴ ± 15.357 (R ² = 0.83) for volume.

Conclusions

With no sex and laterality differences, the fused primary ossification centers of the clavicle grow logarithmically in both transverse and sagittal diameters, linearly in cross-sectional area, and fourth-degree polynomially in volume. Our normative quantitative findings may be conducive in monitoring normal fetal growth and screening for inherited faults and anomalies of the clavicle in European human fetuses.

     

Association of hyperhomocysteinemia with genetic variants in key enzymes of homocysteine metabolism and methotrexate toxicity in rheumatoid arthritis patients

Objectives

The study investigated the association between plasma homocysteine, folate and vitamin B12 with 5,10 methylenetetrahydrofolate reductase (MTHFR C677T and A1298C), thymidylate synthase (TYMS 2R → 3R) and methionine synthase (MTR A2756G) polymorphisms and methotrexate (MTX) treatment and toxicity in Tunisian Rheumatoid arthritis (RA) patients.

Methods

A total of 185 patients with RA were included. Homocysteine (Hcy) was assessed by fluorescence polarization immunoassay, and folate and vitamin B12 were measured by chemiluminescence immunoassays. The genetic polymorphisms were analyzed by PCR or PCR-RFLP. Hyperhomocysteinemia (HHC) was considered for Hcy?>?15 µmol/L.

Results

MTHFR C677T polymorphism was associated with HHC in RA patients (multi-adjusted OR, 95% CI 2.18, [1.07–4.57]; p?=?0.031). No association was detected with the remaining polymorphisms. Plasma Hcy, folate, and vitamin B12 did not differ according to each polymorphism, or with MTX treatment or toxicity. However, HHC was more prevalent in patients with than those without MTX toxicity (32.7 vs. 16.7%; p?=?0.035).

Conclusions

The MTHFR 677TT genotype is an independent risk factor for HHC in Tunisians RA patients. HHC could be a useful marker of MTX toxicity in RA patients.

     

Human NK cells adapt their immune response towards increasing multiplicities of infection of <Emphasis Type="Italic">Aspergillus fumigatus</Emphasis>

Background

The saprophytic fungus Aspergillus fumigatus reproduces by generation of conidia, which are spread by airflow throughout nature. Since humans are inhaling certain amounts of spores every day, the (innate) immune system is constantly challenged. Even though macrophages and neutrophils carry the main burden, also NK cells are regarded to contribute to the antifungal immune response. While NK cells reveal a low frequency, expression and release of immunomodulatory molecules seem to be a natural way of their involvement.

Results

In this study we show, that NK cells secrete chemokines such as CCL3/MIP-1α, CCL4/MIP-1β and CCL5/RANTES early on after stimulation with Aspergillus fumigatus and, in addition, adjust the concentration of chemokines released to the multiplicity of infection of Aspergillus fumigatus.

Conclusions

These results further corroborate the relevance of NK cells within the antifungal immune response, which is regarded to be more and more important in the development and outcome of invasive aspergillosis in immunocompromised patients after hematopoietic stem cell transplantation. Additionally, the correlation between the multiplicity of infection and the expression and release of chemokines shown here may be useful in further studies for the quantification and/or surveillance of the NK cell involvement in antifungal immune responses.

     

Topical ocular dexamethasone decreases intraocular pressure and body weight in rats

Background

Recently, topical dexamethasone-induced ocular hypertension and a consequent loss of retinal ganglion cells (RGCs) have been described in mice. This has been proposed as a model of steroid-induced glaucoma. In this study, we set up and evaluated a similar model in rats.

Results

Ten-week old Sprague Dawley (SD) rats (N?=?12) were used to evaluate the effect of topical 0.1 % dexamethasone (50 μl) administered 3 times daily for 4 weeks. Sodium chloride (0.9 %) was used in another group of rats (N?=?12) that served as the controls. After 1 week, we observed a progressive decrease in body weight in the dexamethasone-treated rats compared both to the pre-treatment baseline and the vehicle-treated rats. In contrast to earlier work that showed elevated Intraocular pressure (IOP) following dexamethasone instillation in mice, IOP in the rats unexpectedly fell to 11.3?±?1.3 mmHg in the treated eyes, compared to 14.8?±?2.4 mmHg in the untreated eyes, after 3 weeks of topical dexamethasone (P?=?0.032). Blood tests performed after 4 weeks of treatment showed a 3.3-fold increase in both plasma cholesterol (P?<?0.001) and alanine transaminase (P?=?0.019) in the dexamethasone-treated rats compared to the control rats. Meanwhile, topical steroid did not induce changes in either plasma blood glucose or glycated hemoglobin (HbA1c). We also did not detect changes in the expression of RGC markers (with real-time PCR) following the treatment.

Conclusions

In contrast to mice, which previously showed increased IOP following the topical administration of dexamethasone, the rats displayed a paradoxical reduction in IOP following a similar treatment. This was accompanied by a loss of body weight without affecting the level of blood glucose.

     

Cognitive-Behavioural Therapy for Inflammatory Bowel Disease: 24-Month Data from a Randomised Controlled Trial

Purpose

There is ongoing controversy on the effectiveness of psychotherapy in inflammatory bowel disease (IBD). In the few small studies, cognitive-behavioural therapy (CBT) has been shown to alleviate symptoms of anxiety or depression. However, there is little research on the impact of CBT on physical outcomes in IBD and no studies on long-term effectiveness of CBT.

Methods

The present two-arm pragmatic randomised controlled trial aimed to establish the impact of CBT on disease course after 24 months of observation. The study compared standard care plus CBT (+CBT) with standard care alone (SC). CBT was delivered over 10 weeks, face-to-face (F2F) or online (cCBT). The data were analysed using linear mixed-effects models.

Results

CBT did not significantly influence disease activity as measured by disease activity indices at 24 months (Crohn’s Disease Activity Index (CDAI), p?=?0.92; Simple Clinical Colitis Activity Index (SCCAI), p?=?0.88) or blood parameters (C-reactive protein (CRP), p?<?0.62; haemoglobin (Hb), p?=?0.77; platelet, p?=?0.64; white cell count (WCC), p?=?0.59) nor did CBT significantly affect mental health, coping or quality of life (all p?>?0.05).

Conclusions

Therefore, we conclude that CBT does not influence the course of IBD over 24 months. Given the high rate of attrition, particularly in the CBT group, future trials should consider a personalised approach to psychotherapy, perhaps combining online and one-to-one therapist time.

     

Endothelial,platelet, and macrophage microparticle levels do not change acutely following transcatheter aortic valve replacement

Background

Patients with severe aortic stenosis have increased levels of prothrombotic and proinflammatory microparticles (MP), and MPs actively regulate pathological processes that lead to atherothrombotic cardiovascular events. Shear stress is a validated stimulus of MP production, and abnormal shear stress in aortic stenosis increases MP release in ex-vivo studies. We hypothesized that in patients with severe aortic stenosis, percutaneous replacement of the aortic valve (TAVR) would reduce abnormal shear stress and would decrease levels of circulating MPs.

Findings

The experimental protocol utilized flow cytometry (FC) and nanoparticle tracking analysis (NTA) to quantify circulating plasma MP levels in aortic stenosis patients at baseline and 5 days after TAVR. The baseline and 5 day MP counts measured by FC were 6.10?10⁵?±?1.21?10⁵ MP/μL and 5.74?10⁵?±?9.54?10⁴ MP/μL, respectively (p?=?0.91). The baseline and 5 day MP counts measured by NTA were 9.29?10¹³?±?1.66?10¹³ MP/μL and 3.95?10¹⁴?±?3.11?10¹⁴ MP/μL, respectively (p?=?0.91). When MPs were stratified by cell source, there was no difference in pre/post TAVR endothelial, platelet, or leukocyte MP levels.

Conclusion

Levels of circulating MPs do not change acutely following TAVR therapy for aortic stenosis.Trial registered at clinicaltrials.gov NCT02193035 on July 11, 2014.