bluuebunny/crossref_metadata_2025 huggingface dataset:
An observational study reveals that neonatal vitamin D is primarily determined by maternal contributions: implications of a new assay on the roles of vitamin D forms
Authors: Karras Spyridon N , Shah Iltaf , Petroczi Andrea , Goulis Dimitrios G , Bili Helen , Papadopoulou Fotini , Harizopoulou Vikentia , Tarlatzis Basil C , Naughton Declan P
Published: June 2013

Background

Vitamin D concentrations during pregnancy are measured to diagnose states of insufficiency or deficiency. The aim of this study is to apply accurate assays of vitamin D forms [single- hydroxylated [25(OH)D2, 25(OH)D3], double-hydroxylated [1α,25(OH)2D2, 1α,25(OH)2D3], epimers [3-epi-25(OH)D2, 3-epi-25(OH)D3] in mothers (serum) and neonates (umbilical cord) to i) explore maternal and neonatal vitamin D biodynamics and ii) to identify maternal predictors of neonatal vitamin D concentrations.

Methods

All vitamin D forms were quantified in 60 mother- neonate paired samples by a novel liquid chromatography -mass spectrometry (LC-MS/MS) assay. Maternal characteristics [age, ultraviolet B exposure, dietary vitamin D intake, calcium, phosphorus and parathyroid hormone] were recorded. Hierarchical linear regression was used to predict neonatal 25(OH)D concentrations.

Results

Mothers had similar concentrations of 25(OH)D2 and 25(OH)D3 forms compared to neonates (17.9 ± 13.2 vs. 15.9 ± 13.6 ng/mL, p = 0.289) with a ratio of 1:3. The epimer concentrations, which contribute approximately 25% to the total vitamin D levels, were similar in mothers and neonates (4.8 ± 7.8 vs. 4.5 ± 4.7 ng/mL, p = 0.556). No correlation was observed in mothers between the levels of the circulating form (25OHD3) and its active form. Neonatal 25(OH)D2 was best predicted by maternal characteristics, whereas 25(OH)D3 was strongly associated to maternal vitamin D forms (R2 = 0.253 vs. 0.076 and R2 = 0.109 vs. 0.478, respectively). Maternal characteristics explained 12.2% of the neonatal 25(OH)D, maternal 25(OH)D concentrations explained 32.1%, while epimers contributed an additional 11.9%.

Conclusions

By applying a novel highly specific vitamin D assay, the present study is the first to quantify 3-epi-25(OH)D concentrations in mother - newborn pairs. This accurate assay highlights a considerable proportion of vitamin D exists as epimers and a lack of correlation between the circulating and active forms. These results highlight the need for accurate measurements to appraise vitamin D status. Maternal characteristics and circulating forms of vitamin D, along with their epimers explain 56% of neonate vitamin D concentrations. The roles of active and epimer forms in the maternal - neonatal vitamin D relationship warrant further investigation.
GP73 is down-regulated in gastric cancer and associated with tumor differentiation
Authors: Chen Le-Gao , Wang Hui-Ju , Yao Hai-bo , Guan Tian-Pei , Wu Fang , He Xu-Jun , Ma Ying-Yu , Tao Hou-Quan , Ye Zai-Yuan
Published: June 2013

Background

Golgi protein 73 (GP73) is a type II Golgi transmembrane protein. It is over-expressed in several cancers, including hepatocellular carcinomas, bile duct carcinomas, lung cancer and prostate cancer. However, there are few reports of GP73 in gastric cancer. This study is aimed at investigating the expression of GP73 and its relationship with clinical pathological characters in gastric cancer.

Methods

GP73 mRNA level was determined by quantitative real-time RT-PCR in 41 pairs of matched gastric tumorous tissues and adjacent non-tumorous mucosal tissues. Western blotting was also performed to detect the GP73 protein level. GP73 protein expression was analyzed by immunohistochemistry in 52 clinically characterized gastric cancer patients and 10 non-tumorous gastric mucosal tissue controls.

Results

The mRNA and protein level of GP73 were significantly down-regulated in gastric tumorous tissues compared with the non-tumorous mucosal tissues. In non-tumorous mucosa, strong diffuse cytoplasmic staining can be seen in cells located at the surface of the glandular and foveolar compartment; while in tumorous tissues, the staining was much weaker or even absent, and mainly in a semi-granular dot-like staining pattern. The expression level of GP73 protein was associated with patients’ gender and tumor differentiation.

Conclusions

GP73 was normally expressed in non-tumorous gastric mucosa and down-regulated in gastric cancer. Its expression in gastric cancer was correlated with tumor differentiation.
Surface roughness effects on the frequency tuning performance of a nanoelectromechanical resonator
Authors: Yoon Hyong Seo , Park Byeongho , Jun Seong Chan
Published: June 2013

Electrothermal heating is one of radio frequency tuning method in nanoelectromechanical resonators with magnetomotive transduction. This study confirmed that the surface roughness of the nanoresonator affects the electrothermal tuning performance under moderate conditions at room temperature. The effect of surface roughness on electrothermal tuning is complicated and involves interactions of mechanical and electrical properties. In addition, the electrothermal damping varied depending on the nanoscale molecular solid structure. These factors affect the signal-to-noise ratio, the effective stress of the beam, and the quality Q-factor of the nanoresonator.
Spatial resolution of confocal XRF technique using capillary optics
Authors: Dehlinger Maël , Fauquet Carole , Lavandier Sebastien , Aumporn Orawan , Jandard Franck , Arkadiev Vladimir , Bjeoumikhov Aniouar , Tonneau Didier
Published: June 2013

XRF (X-ray fluorescence) is a powerful technique for elemental analysis with a high sensitivity. The resolution is presently limited by the size of the primary excitation X-ray beam. A test-bed for confocal-type XRF has been developed to estimate the ultimate lateral resolution which could be reached in chemical mapping using this technique. A polycapillary lens is used to tightly focus the primary X-ray beam of a low power rhodium X-ray source, while the fluorescence signal is collected by a SDD detector through a cylindrical monocapillary. This system was used to characterize the geometry of the fluorescent zone. Capillary radii ranging from 50 μm down to 5 μm were used to investigate the fluorescence signal maximum level This study allows to estimate the ultimate resolution which could be reached in-lab or on a synchrotron beamline. A new tool combining local XRF and scanning probe microscopy is finally proposed.
Absolute flatness measurements of silicon mirrors by a three-intersection method by near-infrared interferometry
Authors: Uchikoshi Junichi , Hayashi Yoshinori , Ajari Noritaka , Kawai Kentaro , Arima Kenta , Morita Mizuho
Published: June 2013

Absolute flatness of three silicon plane mirrors have been measured by a three-intersection method based on the three-flat method using a near-infrared interferometer. The interferometer was constructed using a near-infrared laser diode with a 1,310-nm wavelength light where the silicon plane mirror is transparent. The height differences at the coordinate values between the absolute line profiles by the three-intersection method have been evaluated. The height differences of the three flats were 4.5 nm or less. The three-intersection method using the near-infrared interferometer was useful for measuring the absolute flatness of the silicon plane mirrors.
A cost effective real-time PCR for the detection of adenovirus from viral swabs
Authors: Al-Siyabi Turkiya , Binkhamis Khalifa , Wilcox Melanie , Wong Sallene , Pabbaraju Kanti , Tellier Raymond , Hatchette Todd F , LeBlanc Jason J
Published: June 2013

Compared to traditional testing strategies, nucleic acid amplification tests such as real-time PCR offer many advantages for the detection of human adenoviruses. However, commercial assays are expensive and cost prohibitive for many clinical laboratories. To overcome fiscal challenges, a cost effective strategy was developed using a combination of homogenization and heat treatment with an “in-house” real-time PCR. In 196 swabs submitted for adenovirus detection, this crude extraction method showed performance characteristics equivalent to viral DNA obtained from a commercial nucleic acid extraction. In addition, the in-house real-time PCR outperformed traditional testing strategies using virus culture, with sensitivities of 100% and 69.2%, respectively. Overall, the combination of homogenization and heat treatment with a sensitive in-house real-time PCR provides accurate results at a cost comparable to viral culture.
Cytoplasmic RNA viruses as potential vehicles for the delivery of therapeutic small RNAs
Authors: Usme-Ciro Jose A , Campillo-Pedroza Natalia , Almazán Fernando , Gallego-Gomez Juan C
Published: June 2013

Viral vectors have become the best option for the delivery of therapeutic genes in conventional and RNA interference-based gene therapies. The current viral vectors for the delivery of small regulatory RNAs are based on DNA viruses and retroviruses/lentiviruses. Cytoplasmic RNA viruses have been excluded as viral vectors for RNAi therapy because of the nuclear localization of the microprocessor complex and the potential degradation of the viral RNA genome during the excision of any virus-encoded pre-microRNAs. However, in the last few years, the presence of several species of small RNAs (e.g., virus-derived small interfering RNAs, virus-derived short RNAs, and unusually small RNAs) in animals and cell cultures that are infected with cytoplasmic RNA viruses has suggested the existence of a non-canonical mechanism of microRNA biogenesis. Several studies have been conducted on the tick-borne encephalitis virus and on the Sindbis virus in which microRNA precursors were artificially incorporated and demonstrated the production of mature microRNAs. The ability of these viruses to recruit Drosha to the cytoplasm during infection resulted in the efficient processing of virus-encoded microRNA without the viral genome entering the nucleus. In this review, we discuss the relevance of these findings with an emphasis on the potential use of cytoplasmic RNA viruses as vehicles for the efficient delivery of therapeutic small RNAs.
Placental DNA hypomethylation in association with particulate air pollution in early life
Authors: Janssen Bram G , Godderis Lode , Pieters Nicky , Poels Katrien , Kiciński Michał , Cuypers Ann , Fierens Frans , Penders Joris , Plusquin Michelle , Gyselaers Wilfried , Nawrot Tim S
Published: June 2013

Background

There is evidence that altered DNA methylation is an important epigenetic mechanism in prenatal programming and that developmental periods are sensitive to environmental stressors. We hypothesized that exposure to fine particles (PM2.5) during pregnancy could influence DNA methylation patterns of the placenta.

Methods

In the ENVIRON AGE birth cohort, levels of 5’-methyl-deoxycytidine (5-mdC) and deoxycytidine (dC) were quantified in placental DNA from 240 newborns. Multiple regression models were used to study placental global DNA methylation and in utero exposure to PM2.5 over various time windows during pregnancy.

Results

PM2.5 exposure during pregnancy averaged (25th-75th percentile) 17.4 (15.4-19.3) μg/m3. Placental global DNA methylation was inversely associated with PM2.5 exposures during whole pregnancy and relatively decreased by 2.19% (95% confidence interval [CI]: -3.65, -0.73%, p = 0.004) for each 5 μg/m3 increase in exposure to PM2.5. In a multi-lag model in which all three trimester exposures were fitted as independent variables in the same regression model, only exposure to PM2.5 during trimester 1 was significantly associated with lower global DNA methylation (-2.13% per 5 μg/m3 increase, 95% CI: -3.71, -0.54%, p = 0.009). When we analyzed shorter time windows of exposure within trimester 1, we observed a lower placental DNA methylation at birth during all implantation stages but exposure during the implantation range (6-21d) was strongest associated (-1.08% per 5 μg/m3 increase, 95% CI: -1.80, -0.36%, p = 0.004).

Conclusions

We observed a lower degree of placental global DNA methylation in association with exposure to particulate air pollution in early pregnancy, including the critical stages of implantation. Future studies should elucidate genome-wide and gene-specific methylation patterns in placental tissue that could link particulate exposure during in utero life and early epigenetic modulations.
Infection-associated platelet dysfunction of canine platelets detected in a flow chamber model
Authors: Ferkau Annika , Gillmann Hans-Jörg , Mischke Reinhard , Calmer Simone , Ecklebe Silke , Abid Monia , Minde Jan-Wighard , Echtermeyer Frank , Theilmeier Gregor
Published: June 2013

Background

In the present study, the influence of bacterial infection, lipopolysacharides (LPS) and hydroxyethyl starch (HES) on platelet function in a parallel plate flow chamber were measured. Experiments were performed with non-activated and protease-activating-receptor (PAR) 4 agonist activated platelets. Comparative measurements were in vivo capillary bleeding time, platelet function analyzer and impedance aggregometry.

Results

PAR 4 agonist did not increase platelet adhesion of platelets from dogs with bacterial inflammation in the flow chamber in contrast to platelets of healthy dogs. Except from impedance aggregometry with lower sensitivity and specificity, PFA did not detect platelet dysfunctions in dogs with infection. In vitro addition of LPS or HES significantly reduced platelet covered area after PAR-activation.

Conclusions

The flow chamber detects platelet dysfunctions in dogs with inflammatory diseases. In vitro addition of LPS highlights the inhibiting effect of bacterial wall components on platelet function. Platelet dysfunction induced by infection could possibly also be diagnosed after treatment of sepsis with colloids has commenced. The flow chamber could be a useful tool to detect sepsis associated platelet dysfunction given that larger prospective trials confirm these findings from a proof of concept study.
Acanthamoeba castellanii of the T4 genotype is a potential environmental host for Enterobacter aerogenes and Aeromonas hydrophila
Authors: Yousuf Farzana Abubakar , Siddiqui Ruqaiyyah , Khan Naveed Ahmed
Published: June 2013

Background

Acanthamoeba can interact with a wide range of microorganisms such as viruses, algae, yeasts, protists and bacteria including Legionella pneumophila, Pseudomonas aeruginosa, Vibrio cholerae, Helicobacter pylori, Listeria monocytogenes, Mycobacterium spp., and Escherichia coli. In this capacity, Acanthamoeba has been suggested as a vector in the transmission of bacterial pathogens to the susceptible hosts.

Methods

Here, we used a keratitis isolate of A. castellanii of the T4 genotype and studied its interactions with two bacterial genera which have not been tested before, Enterobacter aerogenes, and Aeromonas hydrophila, as well as E. coli. Assays were performed to determine bacterial association with and invasion of A. castellanii. Additionally, bacterial survival intracellular of A. castellanii trophozoites as well as cysts was determined.

Results

All three bacterial isolates tested, associated, invaded, and survived inside A. castellanii trophozoites as well as A. castellanii cysts. However, E. aerogenes and E. coli exhibited significantly reduced association with and invasion of A. castellanii as compared with A. hydrophila (P < 0.01 using paired T-test, one tail distribution). In the long term survival assays, all three bacterial isolates tested remained viable inside A. castellanii trophozoites, while amoeba remained intact; however A. hydrophila exhibited higher survival inside amoebae (14.54 ± 3.3 bacteria:amoeba ratio) compared with E. aerogenes (3.96 ± 0.7 bacteria:amoeba ratio) and E. coli (5.85 ± 1.1 bacteria:amoeba ratio). A. hydrophila, E. coli, and E. aerogenes remained viable during the encystment process and exhibited higher levels of recovery from mature cysts (14.13 ± 0.89 A. hydrophila:amoeba ratio, 10.13 ± 1.17 E. aerogenes:amoeba ratio, and 11.95 ± 0.7 E. coli:amoeba ratio).

Conclusions

A. hydrophila and E. aerogenes also joined the ranks of other bacteria that could benefit from A. castellanii. Because cysts can be airborne, these findings suggest that Acanthamoeba is a potential vector in the transmission of A. hydrophila and E. aerogenes to susceptible hosts.
Synthesis, docking and acetylcholinesterase inhibitory assessment of 2-(2-(4-Benzylpiperazin-1-yl)ethyl)isoindoline-1,3-dione derivatives with potential anti-Alzheimer effects
Authors: Mohammadi-Farani Ahmad , Ahmadi Aram , Nadri Hamid , Aliabadi Alireza
Published: June 2013

Background

Alzheimer’s disease (AD) as neurodegenerative disorder, is the most common form of dementia accounting for about 50-60% of the overall cases of dementia among persons over 65 years of age. Low acetylcholine (ACh) concentration in hippocampus and cortex areas of the brain is one of the main reasons for this disease. In recent years, acetylcholinesterase (AChE) inhibitors like donepezil with prevention of acetylcholine hydrolysis can enhance the duration of action of acetylcholine in synaptic cleft and improve the dementia associated with Alzheimer’s disease.

Results

Design, synthesis and assessment of anticholinesterase activity of 2-(2-(4-Benzylpiperazin-1-yl)ethyl)isoindoline-1,3-dione derivatives showed prepared compounds can function as potential acetylcholinesterase inhibitor. Among 12 synthesized derivatives, compound 4a with ortho chlorine moiety as electron withdrawing group exhibited the highest potency in these series (IC50 = 0.91 ± 0.045 μM) compared to donepezil (IC50 = 0.14 ± 0.03 μM). The results of the enzyme inhibition test (Ellman test) showed that electron withdrawing groups like Cl, F and NO2 can render the best effect at position ortho and para of the phenyl ring. But compound 4g with methoxy group at position 3(meta) afforded a favorable potency (IC50 = 5.5 ± 0.7 μM). Furthermore, docking study confirmed a same binding mode like donepezil for compound 4a.

Conclusions

Synthesized compounds 4a-4l could be proposed as potential anticholinesterase agents.
Idiopathic inflammatory myopathies: pathogenic mechanisms of muscle weakness
Authors: Rayavarapu Sree , Coley William , Kinder Travis B , Nagaraju Kanneboyina
Published: June 2013

Idiopathic inflammatory myopathies (IIMs) are a heterogenous group of complex muscle diseases of unknown etiology. These diseases are characterized by progressive muscle weakness and damage, together with involvement of other organ systems. It is generally believed that the autoimmune response (autoreactive lymphocytes and autoantibodies) to skeletal muscle-derived antigens is responsible for the muscle fiber damage and muscle weakness in this group of disorders. Therefore, most of the current therapeutic strategies are directed at either suppressing or modifying immune cell activity. Recent studies have indicated that the underlying mechanisms that mediate muscle damage and dysfunction are multiple and complex. Emerging evidence indicates that not only autoimmune responses but also innate immune and non-immune metabolic pathways contribute to disease pathogenesis. However, the relative contributions of each of these mechanisms to disease pathogenesis are currently unknown. Here we discuss some of these complex pathways, their inter-relationships and their relation to muscle damage in myositis. Understanding the relative contributions of each of these pathways to disease pathogenesis would help us to identify suitable drug targets to alleviate muscle damage and also improve muscle weakness and quality of life for patients suffering from these debilitating muscle diseases.
Identification, technological and safety characterization of Lactobacillus sakei and Lactobacillus curvatus isolated from Argentinean anchovies (Engraulis anchoita)
Authors: Belfiore Carolina , Raya Raúl R , Vignolo Graciela M
Published: June 2013

In this study, the identification and characterization of Lactobacillus previously isolated from fresh anchovies (Engraulis anchoita) are investigated. 16S rDNA partial sequencing assigned all the isolates to belong to the Lactobacillus sakei/curvatus group. Fourteen out of 15 isolates were identified as L. sakei by phenotypic traits: they exhibited catalase activity and fermented melibiose, although only 10 of them hydrolyzed arginine. These results were confirmed by multiplex PCR-based restriction enzyme analysis with Hin dIII and by restriction fragment length polymorphic (RFLP) analysis of the 16S-23S rDNA intergenic spacer region with Taq I. Among identified isolates, four L. sakei strains and the sole L. curvatus strain showing sensitivity to chloramphenicol, erythromycin and tetracycline and exhibiting high tolerance to NaCl (10-18%) were unable to produce neither dextran nor biogenic amines. Based on technological and safety features, L. sakei SACB704 and L. curvatus SACB03a naturally present in fresh anchovies may be promising strains for the development of a starter culture to accelerate and control the fermentation of salt fermented anchovy-based products.
Seaweed-mediated synthesis of gold nanoparticles using Turbinaria conoides and its characterization
Authors: Rajeshkumar Shanmugam , Malarkodi Chelladurai , Gnanajobitha Gnanadhas , Paulkumar Kanniah , Vanaja Mahendran , Kannan Chellapandian , Annadurai Gurusamy
Published: June 2013

The synthesis of metal nanoparticles using algae has been unexplored, but it is a more biocompatible method than the other biological methods. Metal nanoparticle synthesis using algae extract shows rapid and non-toxic process which resulted to nano sizes having the greatest potential for biomedical applications. In this investigation, we studied the green synthesis of gold nanoparticles using the algae extract of Turbinaria conoides. Green synthesis of gold nanoparticles was preliminarily confirmed by color changing from yellow to dark pink in the reaction mixture, and the broad surface plasmon resonance band was centered at 520 to 525 nm which indicates polydispersed nanoparticles. Transmission electron microscopy and selected-area electron diffraction analysis show the morphology and crystalline structure of synthesized gold nanoparticles with the size range of 6 to 10 nm. The four strong diffraction peaks were observed by X-ray diffraction; it confirmed the crystalline nature of synthesized gold nanoparticles. The carboxylic, amine, and polyphenolic groups were associated with the algae-assisted synthesized gold nanoparticles which was confirmed using Fourier transform-infrared spectroscopy. This study eliminates the use of chemical substances as reducing and stabilizing agent. Because it has natural several constituents which are fucoidan and polyphenolic substances, it does a dual function as both reducing and stabilizing agent for nanoparticles. Thus, algae-mediated synthesis process of biomedically valuable gold nanoparticles is a one-spot, facile, convenient, large-scaled, and eco-friendly method.
Human conditionally immortalized neural stem cells improve locomotor function after spinal cord injury in the rat
Authors: Amemori Takashi , Romanyuk Nataliya , Jendelova Pavla , Herynek Vit , Turnovcova Karolina , Prochazka Pavel , Kapcalova Miroslava , Cocks Graham , Price Jack , Sykova Eva
Published: June 2013

Introduction

A growing number of studies have highlighted the potential of stem cell and more-differentiated neural cell transplantation as intriguing therapeutic approaches for neural repair after spinal cord injury (SCI).

Methods

A conditionally immortalized neural stem cell line derived from human fetal spinal cord tissue (SPC-01) was used to treat a balloon-induced SCI. SPC-01 cells were implanted into the lesion 1 week after SCI. To determine the feasibility of tracking transplanted stem cells, a portion of the SPC-01 cells was labeled with poly-L-lysine-coated superparamagnetic iron-oxide nanoparticles, and the animals grafted with labeled cells underwent magnetic resonance imaging. Functional recovery was evaluated by using the BBB and plantar tests, and lesion morphology, endogenous axonal sprouting and graft survival, and differentiation were analyzed. Quantitative polymerase chain reaction (qPCR) was used to evaluate the effect of transplanted SPC-01 cells on endogenous regenerative processes.

Results

Transplanted animals displayed significant motor and sensory improvement 2 months after SCI, when the cells robustly survived in the lesion and partially filled the lesion cavity. qPCR revealed the increased expression of rat and human neurotrophin and motor neuron genes. The grafted cells were immunohistologically positive for glial fibrillary acidic protein (GFAP); however, we found 25% of the cells to be positive for Nkx6.1, an early motor neuron marker. Spared white matter and the robust sprouting of growth-associated protein 43 (GAP43)+ axons were found in the host tissue. Four months after SCI, the grafted cells matured into Islet2+ and choline acetyltransferase (ChAT)+ neurons, and the graft was grown through with endogenous neurons. Grafted cells labeled with poly-L-lysine-coated superparamagnetic nanoparticles before transplantation were detected in the lesion on T2-weighted images as hypointense spots that correlated with histologic staining for iron and the human mitochondrial marker MTCO2.

Conclusions

The transplantation of SPC-01 cells produced significant early functional improvement after SCI, suggesting an early neurotrophic action associated with long-term restoration of the host tissue, making the cells a promising candidate for future cell therapy in patients with SCI.
B7-1/B7-2 blockade overrides the activation of protective CD8 T cells stimulated in the absence of Foxp3+ regulatory T cells
Authors: Ertelt James M , Buyukbasaran Esra Z , Jiang Tony T , Rowe Jared H , Xin Lijun , Way Sing Sing
Published: June 2013

Although T cell activation has been classically described to require distinct, positive stimulation signals that include B7-1 (CD80) and B7-2 (CD86) costimulation, overriding suppression signals that avert immune-mediated host injury are equally important. How these opposing stimulation and suppression signals work together remains incompletely defined. Our recent studies demonstrate that CD8 Teff activation in response to cognate peptide stimulation is actively suppressed by the Foxp3+ subset of CD4 cells, called Tregs. Here, we show that the elimination of Treg suppression does not bypass the requirement for positive B7-1/B7-2 costimulation. The expansion, IFN-γ cytokine production, cytolytic, and protective features of antigen-specific CD8 T cells stimulated with purified cognate peptide in Treg-ablated mice were each neutralized effectively by CTLA-4-Ig that blocks B7-1/B7-2. In turn, given the efficiency whereby CTLA-4-Ig overrides the effects of Treg ablation, the role of Foxp3+ cell-intrinsic CTLA-4 in mitigating CD8 Teff activation was also investigated. With the use of mixed chimera mice that contain CTLA-4-deficient Tregs exclusively after the ablation of WT Foxp3+ cells, a critical role for Treg CTLA-4 in suppressing the expansion, cytokine production, cytotoxicity, and protective features of peptide-stimulated CD8 T cells is revealed. Thus, the activation of protective CD8 T cells requires positive B7-1/B7-2 costimulation even when suppression by Tregs and in particular, Treg-intrinsic CTLA-4 is circumvented.
RIG-I activation inhibits HIV replication in macrophages
Authors: Wang Yizhong , Wang Xu , Li Jieliang , Zhou Yu , Ho Wenzhe
Published: June 2013

The RIG-I signaling pathway is critical in the activation of the type I IFN-dependent antiviral innate-immune response. We thus examined whether RIG-I activation can inhibit HIV replication in macrophages. We showed that the stimulation of monocyte-derived macrophages with 5′ppp-dsRNA, a synthetic ligand for RIG-I, induced the expression of RIG-I, IFN-α/β, and several IRFs, key regulators of the IFN signaling pathway. In addition, RIG-I activation induced the expression of multiple intracellular HIV-restriction factors, including ISGs, several members of the APOBEC3 family, tetherin and CC chemokines, the ligands for HIV entry coreceptor (CCR5). The inductions of these factors were associated with the inhibition of HIV replication in macrophages stimulated by 5′ppp-dsRNA. These observations highlight the importance of RIG-I signaling in macrophage innate immunity against HIV, which can be beneficial for the treatment of HIV disease, where intracellular immune defense is compromised by the virus.
MGL signaling augments TLR2-mediated responses for enhanced IL-10 and TNF-α secretion
Authors: van Vliet Sandra J , Bay Sylvie , Vuist Ilona M , Kalay Hakan , García-Vallejo Juan J , Leclerc Claude , van Kooyk Yvette
Published: June 2013

DCs orchestrate immune responses to infectious pathogens and disturbances in tissue integrity. Equipped with C-type lectins, DCs can respond to environmental changes in glycosylation. Many C-type lectins are capable of modulating TLR activation, thereby facilitating tailor-made immune reactions. Here, we investigated the signaling properties of the C-type lectin MGL and show that MGL engagement by agonistic antibodies or carbohydrate ligands couples to TLR signal transduction for increased IL-10 and TNF-α secretion by human monocyte-derived DCs. MGL triggering especially synergized with TLR2-induced pathways, leading to elevated IL-10 mRNA levels and enhanced TNF-α mRNA stability. In addition, MGL signaling promoted phosphorylation of the MAPK ERK and the transcription factor CREB. Whereas specific inhibitors of p90RSK blocked the MGL-induced cytokine secretion, AP-1 was not involved. Strikingly, NF-κB was only crucial for the IL-10 response and dispensable for TNF-α production. Together, our results demonstrate that MGL activation of the ERK-p90RSK-CREB axis converges with TLR2-induced pathways, thereby fine-tuning the DC maturation phenotype.
Inhibition of geranylgeranyltransferase attenuates neutrophil accumulation and tissue injury in severe acute pancreatitis
Authors: Merza Mohammed , Wetterholm Erik , Zhang Su , Regner Sara , Thorlacius Henrik
Published: June 2013

Leukocyte infiltration and acinar cell necrosis are hallmarks of severe AP, but the signaling pathways regulating inflammation and organ injury in the pancreas remain elusive. In the present study, we investigated the role of geranylgeranyltransferase in AP. Male C57BL/6 mice were treated with a geranylgeranyltransferase inhibitor GGTI-2133 (20 mg/kg) prior to induction of pancreatitis by infusion of taurocholate into the pancreatic duct. Pretreatment with GGTI-2133 reduced plasma amylase levels, pancreatic neutrophil recruitment, hemorrhage, and edema formation in taurocholate-evoked pancreatitis. Moreover, administration of GGTI-2133 decreased the taurocholate-induced increase of MPO activity in the pancreas and lung. Treatment with GGTI-2133 markedly reduced levels of CXCL2 in the pancreas and IL-6 in the plasma in response to taurocholate challenge. Notably, geranylgeranyltransferase inhibition abolished neutrophil expression of Mac-1 in mice with pancreatitis. Finally, inhibition of geranylgeranyltransferase had no direct effect on secretagogue-induced activation of trypsinogen in pancreatic acinar cells in vitro. A significant role of geranylgeranyltransferase was confirmed in an alternate model of AP induced by L-arginine challenge. Our findings show that geranylgeranyltransferase regulates neutrophil accumulation and tissue damage via expression of Mac-1 on neutrophils and CXCL2 formation in AP. Thus, these results reveal new signaling mechanisms in pancreatitis and indicate that targeting geranylgeranyltransferase might be an effective way to ameliorate severe AP.
Childhood maltreatment and methylation of the glucocorticoid receptor geneNR3C1in bipolar disorder
Authors: Perroud Nader , Dayer Alexandre , Piguet Camille , Nallet Audrey , Favre Sophie , Malafosse Alain , Aubry Jean-Michel
Published: June 2013

Background

Early-life adversities represent risk factors for the development of bipolar affective disorder and are associated with higher severity of the disorder. This may be the consequence of a sustained alteration of the hypothalamic–pituitary–adrenal (HPA) axis resulting from epigenetic modifications of the gene coding for the glucocorticoid receptor (NR3C1).

Aims

To investigate whether severity of childhood maltreatment is associated with increased methylation of the exon? 1FNR3C1promoter in bipolar disorder.

Method

A sample of people with bipolar disorder (n= 99) were assessed for childhood traumatic experiences. The percentage ofNR3C1methylation was measured for each participant.

Results

The higher the number of trauma events, the higher was the percentage ofNR3C1methylation (β = 0.52, 95% CI 0.46–0.59,P<<0.0001). The severity of each type of maltreatment (sexual, physical and emotional) was also associated withNR3C1methylation status.

Conclusions

Early-life adversities have a sustained effect on the HPA axis through epigenetic processes and this effect may be measured in peripheral blood. This enduring biological impact of early trauma may alter the development of the brain and lead to adult psychopathological disorder.
Characteristics and rates of mental health problems among Indian and White adolescents in two English cities
Authors: Dogra Nisha , Svirydzenka Nadzeya , Dugard Pat , Singh Swaran P. , Vostanis Panos
Published: June 2013

Background

Sampling techniques for national surveys have constrained the statistical power in estimating prevalence rates of child mental health problems in minority ethnic groups.

Aims

To establish the prevalence rates of mental health problems in ethnic Indian adolescents in England and compare these with matched White adolescents living in the same areas.

Method

A cross-sectional survey with oversampling of Indian adolescents aged 13–15 years of age.

Results

The sample size was 2900 (71% response rate) with 1087 (37%) Indian and 414 (14%) White adolescents. Ethnically Indian adolescents had lower rates of all types of mental

health problems (5% v. 13% and 21% v. 30% for abnormal Strengths and Difficulties Questionnaire and Short Mood and Feelings Questionnaire scores, respectively) and substance misuse (18% v. 57%, 5% v. 15% and 6% v. 9% for regular alcohol, smoking and drug use, respectively), with the exception of eating disorders, compared with their White counterparts. The odds of an abnormal score on the mental health questionnaires were worse for White compared with Indian children irrespective of sociodemographic variables.

Conclusions

Factors relating to how Indian adolescents are parented or their social support networks may be influencing their mental health and may warrant further investigation.
Kisspeptin Activation of TRPC4 Channels in Female GnRH Neurons Requires PIP2 Depletion and cSrc Kinase Activation
Authors: Zhang Chunguang , Bosch Martha A. , Rønnekleiv Oline K. , Kelly Martin J.
Published: June 2013

Kisspeptin signaling via its Gαq-coupled receptor GPR54 plays a crucial role in modulating GnRH neuronal excitability, which controls pituitary gonadotropins secretion and ultimately reproduction. Kisspeptin potently depolarizes GnRH neurons primarily through the activation of canonical transient receptor potential (TRPC) channels, but the intracellular signaling cascade has not been elucidated. Presently, we have established that kisspeptin activation of TRPC channels requires multiple membrane and intracellular signaling molecules. First, phosphatidylinositol-4,5-bisphosphate (PIP2) hydrolysis by phospholipase Cβ is required because whole-cell dialysis of Dioctanoylglycerol-PIP2 (DiC8-PIP2) inhibited the kisspeptin activation of TRPC channels, and the phosphatidylinositol 4-kinase inhibitor wortmannin, which attenuates PIP2 synthesis, prolonged TRPC channel activation. Using single cell RT-PCR, we identified that the mRNA for the PIP2-interacting TRPC channel subunit, TRPC4α, is expressed in GnRH neurons. Depletion of intracellular Ca2+ stores by thapsigargin and inositol 1,4,5-trisphosphate had no effect, indicating that the TRPC channels are not store-operated. Neither removing extracellular Ca2+ nor buffering intracellular Ca2+ with EGTA or BAPTA had any effect on the kisspeptin activation of the TRPC channels. However, the Ca2+ channel blocker Ni2+ inhibited the kisspeptin-induced inward current. Moreover, inhibition of protein kinase C by bisindolylmaleimide-I or calphostin C had no effect, but activation of protein kinase C by phorbol 12,13-dibutyrate occluded the kisspeptin-activated current. Finally, inhibition of the cytoplasmic tyrosine kinase cSrc by genistein or the pyrazolo-pyrimidine PP2 blocked the activation of TRPC channels by kisspeptin. Therefore, TRPC channels in GnRH neurons are receptor-operated, and kisspeptin activates TRPC channels through PIP2 depletion and cSrc tyrosine kinase activation, which is a novel signaling pathway for peptidergic excitation of GnRH neurons.
Regulation of Arcuate Neurons Coexpressing Kisspeptin, Neurokinin B, and Dynorphin by Modulators of Neurokinin 3 and κ-Opioid Receptors in Adult Male Mice
Authors: Ruka Kristen A. , Burger Laura L. , Moenter Suzanne M.
Published: June 2013

Pulsatile GnRH release is essential to fertility and is modulated by gonadal steroids, most likely via steroid-sensitive afferents. Arcuate neurons coexpressing kisspeptin, neurokinin B (NKB), and dynorphin (KNDy neurons) are steroid-sensitive and have been postulated to both generate GnRH pulses and mediate steroid feedback on pulse frequency. KNDy neurons are proposed to interact with one another via NKB and dynorphin to activate and inhibit the KNDy network, respectively, and thus alter kisspeptin output to GnRH neurons. To test the roles of NKB and dynorphin on KNDy neurons and the steroid sensitivity of these actions, targeted extracellular recordings were made of Tac2(NKB)-GFP-identified neurons from castrate and intact male mice. Single-cell PCR confirmed most of these cells had a KNDy phenotype. The neurokinin 3 receptor (NK3R) agonist senktide increased action potential firing activity of KNDy neurons. Dynorphin reduced spontaneous KNDy neuron activity, but antagonism of κ-opioid receptors (KOR) failed to induce firing activity in quiescent KNDy neurons. Senktide-induced activation was greater in KNDy neurons from castrate mice, whereas dynorphin-induced suppression was greater in KNDy neurons from intact mice. Interactions of dynorphin with senktide-induced activity were more complex; dynorphin treatment after senktide had no consistent inhibitory effect, whereas pretreatment with dynorphin decreased senktide-induced activity only in KNDy neurons from intact but not castrate mice. These data suggest dynorphin-mediated inhibition of senktide-induced activity requires gonadal steroid feedback. Together, these observations support the hypotheses that activation of NK3R and KOR, respectively, excites and inhibits KNDy neurons and that gonadal steroids modulate these effects.
Minireview: Dopaminergic Regulation of Insulin Secretion from the Pancreatic Islet
Authors: Ustione Alessandro , Piston David W. , Harris Paul E.
Published: June 2013

Exogenous dopamine inhibits insulin secretion from pancreatic β-cells, but the lack of dopaminergic neurons in pancreatic islets has led to controversy regarding the importance of this effect. Recent data, however, suggest a plausible physiologic role for dopamine in the regulation of insulin secretion. We review the literature underlying our current understanding of dopaminergic signaling that can down-regulate glucose-stimulated insulin secretion from pancreatic islets. In this negative feedback loop, dopamine is synthesized in the β-cells from circulating l-dopa, serves as an autocrine signal that is cosecreted with insulin, and causes a tonic inhibition on glucose-stimulated insulin secretion. On the whole animal scale, l-dopa is produced by cells in the gastrointestinal tract, and its concentration in the blood plasma increases following a mixed meal. By reviewing the outcome of certain types of bariatric surgery that result in rapid amelioration of glucose tolerance, we hypothesize that dopamine serves as an “antiincretin” signal that counterbalances the stimulatory effect of glucagon-like peptide 1.
CEH-20/Pbx and UNC-62/Meis function upstream of rnt-1/Runx to regulate asymmetric divisions of the C. elegans stem-like seam cells
Authors: Hughes Samantha , Brabin Charles , Appleford Peter J. , Woollard Alison
Published: June 2013

Caenorhabditis elegans seam cells divide in the stem-like mode throughout larval development, with the ability to both self-renew and produce daughters that differentiate. Seam cells typically divide asymmetrically, giving rise to an anterior daughter that fuses with the hypodermis and a posterior daughter that proliferates further. Previously we have identified rnt-1 (a homologue of the mammalian cancer-associated stem cell regulator Runx) as being an important regulator of seam development, acting to promote proliferation; rnt-1 mutants have fewer seam cells whereas overexpressing rnt-1 causes seam cell hyperplasia. We isolated the interacting CEH-20/Pbx and UNC-62/Meis TALE-class transcription factors during a genome-wide RNAi screen for novel regulators of seam cell number. Animals lacking wild type CEH-20 or UNC-62 display seam cell hyperplasia, largely restricted to the anterior of the worm, whereas double mutants have many additional seam cells along the length of the animal. The cellular basis of the hyperplasia involves the symmetrisation of normally asymmetric seam cell divisions towards the proliferative stem-like fate. The hyperplasia is completely suppressed in rnt-1 mutants, and rnt-1 is upregulated in ceh-20 and unc-62 mutants, suggesting that CEH-20 and UNC-62 function upstream of rnt-1 to limit proliferative potential to the appropriate daughter cell. In further support of this we find that CEH-20 is asymmetrically localised in seam daughters following an asymmetric division, being predominantly restricted to anterior nuclei whose fate is to differentiate. Thus, ceh-20 and unc-62 encode crucial regulators of seam cell division asymmetry, acting via rnt-1 to regulate the balance between proliferation and differentiation.
Receptor guanylyl cyclase Gyc76C is required for invagination, collective migration and lumen shape in the Drosophila embryonic salivary gland
Authors: Patel Unisha , Myat Monn Monn
Published: June 2013

The Drosophila embryonic salivary gland is formed by the invagination and collective migration of cells. Here, we report on a novel developmental role for receptor-type guanylyl cyclase at 76C, Gyc76C, in morphogenesis of the salivary gland. We demonstrate that Gyc76C and downstream cGMP-dependent protein kinase 1 (DG1) function in the gland and surrounding mesoderm to control invagination, collective migration and lumen shape. Loss of gyc76C resulted in glands that failed to invaginate, complete posterior migration and had branched lumens. Salivary gland migration defects of gyc76C mutant embryos were rescued by expression of wild-type gyc76C specifically in the gland or surrounding mesoderm, whereas invagination defects were rescued primarily by expression in the gland. In migrating salivary glands of gyc76C mutant embryos, integrin subunits localized normally to gland–mesoderm contact sites but talin localization in the surrounding circular visceral mesoderm and fat body was altered. The extracellular matrix protein, laminin, also failed to accumulate around the migrating salivary gland of gyc76C mutant embryos, and gyc76C and laminin genetically interacted in gland migration. Our studies suggest that gyc76C controls salivary gland invagination, collective migration and lumen shape, in part by regulating the localization of talin and the laminin matrix.
Concentration of isoprostanes as a marker reflecting oxidative status in moderate dyslipidemia
Authors: Hałabiś Magdalena , Kimak Elżbieta , Solski Janusz
Published: June 2013

Dyslipidemia and oxidative stress are considered one of the major causes which lead to many diseases, especially atherosclerosis development, cardiovascular disorders and aggravate risk for cancers, aging and others. Lipid hydroperoxides are the initial products when lipids are damaged by free radicals and in plasma HDL is their major carrier. F2-IsoP are stable compounds and specific products of lipid peroxidation only in phospholipids in vivo through nonenzymatic free-radical mechanisms. Also, they are found in detectable quantities esterified in all biological tissues and in free form in all normal biological fluids. Quantifying of them is considered as accurate way to measure oxidative stress in vivo and could be used as an indicator of lipid peroxidation which process is a risk factor for atherosclerosis and other systemic diseases. Our study was performed in the group of young people (n=10, age 19-30) with moderate dyslipidemia and the reference group. In both groups we have investigated serum lipids (TG, TC, LDL-C, HDL-C, nonHDL-C), lipoproteins (apoAI, apoB) and lipid (TC/HDL-C, TG/HDL-C, LDL-C/HDL-C) and lipoprotein (HDL-C/apoAI) ratios and oxidative status by measuring free and total (free and esterified) F2-isoprostanes (F2-IsoP). According to the available data reports and taking into account results obtained in our study we concluded that even mild lipid and lipoprotein abnormalities which are often present in population might by connected with elevated oxidative status which is sensitively reflected by F2-IsoP concentration. Thus measurement of F2-IsoP seems to be very helpful in recognition of early stage of oxidative stress however further studies are required.
Influence of extraction conditions on the recovery of the sum of phenolic compounds from daily food rations of students
Authors: Koch Wojciech , Kukuła-Koch Wirginia , Marzec Zbigniew , Jurek Marta
Published: June 2013

The aim of this study was to determine the influence of the applied extraction techniques in conjunction with the selection of an appropriate solvent system in the recovery of phenolic compounds from daily food rations of students (DFR). The study was performed on the averaged diets reconstructed on the basis of 24-dietary recall conducted among 162 randomly chosen people. It was shown that the most effective extraction technique was found to be the accelerated solvent extraction and the most optimal extrahents: mixture of acetone and water in a volume ratio of 7:3.
The level of patient satisfaction with implant treatment of missing teeth
Authors: Koszuta Agnieszka , Szymańska Jolanta ,   Chair and Department of Paedodontics, Medical University of Lublin
Published: June 2013

The aim of the study was to evaluate the level of patient satisfaction with implant treatment of missing teeth. The survey was conducted among 464 patients of both sexes aged from 20 to 74 years, treated with dental implants at the Non-Public Healthcare Centre “Dental” in Tomaszów Mazowiecki. The results of the study reveal that out of the half of all the patients treated with temporary dental prostheses, 71.67% were satisfied; a majority of the latter group were patients younger than 40 years, while the respondents aged over 60 were least frequently satisfied. A great majority of patients assessed other evaluated aspects of implant-prosthetic therapy positively. A high proportion of “highly satisfied” (strongly agree) and “somewhat satisfied” (“agree”) answers, reaching 95%, concerned the shape and colour of implant-supported prostheses and the outcome of the therapy. The smallest group of the respondents observed speech improvement as a result of implant-prosthetic treatment (15.51%). The results of the survey indicate that treatment of missing teeth with dental implants meets patients’ expectations to a high degree. Prosthetic therapy with implants is a method that allows a fully functional and aesthetic treatment of missing teeth.