bluuebunny/crossref_metadata_2025 huggingface dataset:
Landmark advances in the development of erythropoietin
Authors: Fisher James W
Published: April 2011

This is a Minireview covering landmarks or milestones in the development of erythropoietin (EPO). Thirty-nine landmark advances have been identified, which cover the period 1863–2003. Several reports are included that directly support these original landmark advances. This Minireview also updates some of the advances in EPO research since my last Minireview update on EPO published in this journal in 2003. The areas of EPO research updated are: sites of production; purification, assay and standardization; regulation; action; use in anemias; extraerythropoietic actions; adverse effects; and blood doping. The new reports on the use of EPO in the therapy of myocardial infarction; stroke and other neurological diseases; diabetic retinopathy and other retinal diseases are also covered.
Haptoglobin as an early serum biomarker of virus-induced autoimmune type 1 diabetes in biobreeding diabetes resistant and LEW1.WR1 rats
Authors: Kruger Annie J , Yang Chaoxing , Tam Sun W , Hinerfeld Douglas , Evans James E , Green Karin M , Leszyk John , Yang Kejian , Guberski Dennis L , Mordes John P , Greiner Dale L , Rossini Aldo A , Bortell Rita
Published: April 2011

Proteomic profiling of serum is a powerful technique to identify differentially expressed proteins that can serve as biomarkers predictive of disease onset. In this study, we utilized two-dimensional (2D) gel analysis followed by matrix-assisted-laser desorption/ionization time-of-flight mass spectrometry analysis to identify putative serum biomarkers for autoimmune type 1 diabetes (T1D) in biobreeding diabetes resistant (BBDR) rats induced to express the disease. Treatment with toll-like receptor 3 ligand, polyinosinic:polycytidilic acid (pIC), plus infection with Kilham rat virus (KRV), a rat parvovirus, results in nearly 100% of young BBDR rats becoming diabetic within 11–21 d. Sera collected from prediabetic rats at early time points following treatment with pIC + KRV were analyzed by 2D gel electrophoresis and compared with sera from control rats treated with phosphate-buffered saline, pIC alone or pIC + H1, a non-diabetogenic parvovirus. None of the latter three control treatments precipitates T1D. 2D gel analysis revealed that haptoglobin, an acute phase and hemoglobin scavenger protein, was differentially expressed in the sera of rats treated with pIC + KRV relative to control groups. These results were confirmed by Western blot and enzyme-linked immunosorbent assay studies, which further validated haptoglobin levels as being differentially increased in the sera of pIC + KRV-treated rats relative to controls during the first week following infection. Early elevations in serum haptoglobin were also observed in LEW1.WR1 rats that became diabetic following infection with rat cytomegalovirus. The identification and validation of haptoglobin as a putative serum biomarker for autoimmune T1D in rats now affords us the opportunity to test the validity of this protein as a biomarker for human T1D, particularly in those situations where viral infection is believed to precede the onset of disease.
Persistent enteric mycobacterial infection enhances sensitivity to acute mucosal injury
Authors: Johnson Charles S , Wannemuehler Michael J , Hostetter Jesse M
Published: April 2011

Antigen presenting cells such as intestinal macrophages are dynamic effector cells that play a critical role in maintaining mucosal homeostasis. However, it is not known how occult intestinal infections alter the response of the intestinal mucosa to subsequent intestinal injury. The aim of this study was to evaluate how persistent subclinical intestinal infection with Mycobacterium avium subsp. paratuberculosis ( Map) would influence acute dextran sulfate sodium (DSS)-mediated intestinal inflammation. BALB/c mice were infected intraperitoneally with Map. Following an incubation period of 90 d, mice were administered 2% DSS in the drinking water for six days. Prior to and during treatment with DSS, mice were evaluated for clinical signs of disease and body weights were recorded. At termination of the experiment, body weights, frequency of rectal blood, and gross and histological cecal lesions were evaluated, and tissues were collected for isolation of Map. Subclinical and persistent intestinal Map infection was established based on the absence of both weight loss and rectal blood and the isolation of Map from the small and large intestines in mice infected with Map only. Following treatment with DSS, Map-infected mice had increased weight loss, increased frequency of rectal blood, and exacerbation of gross lesions and increased cecal lesion scores. Also, there was a significant reduction in Map isolated from the small intestines of Map-infected and DSS-treated mice. In conclusion, subclinical Map infection sensitizes the host to enhanced acute DSS-mediated intestinal inflammation.
Motor and behavioral changes in rats with adenine-induced chronic renal failure: influence of acacia gum treatment
Authors: Ali Badreldin H , Ziada Amal , Al Husseni Isehaq , Beegam Sumaya , Nemmar Abderrahim
Published: April 2011

Chronic renal failure (CRF) either occurring naturally in humans or induced surgically in rats causes alterations in behavior and motor functions. However, the effect of chemically induced CRF in rats on behavior is not known. We induced CRF in rats by feeding adenine (0.75% w/w, four weeks) and investigated the effect of the ensuing CRF on a depression model (forced swimming test, FST), analgesia (mechanical nociception), neuromuscular coordination (Rota-rod test) and motor activity (activity meter test). Further, we investigated the effect of giving acacia gum (AG, 10% w/v) in the drinking water concomitantly with adenine using the above models. AG has been previously shown to ameliorate the severity of CRF in humans and rats. Adenine-induced CRF significantly increased the plasma concentrations of urea and creatinine, and reduced creatinine clearance. Additionally, it significantly reduced motor activity and increased immobility time in the FST, suggesting a depressant-like effect. Both of these actions were significantly antagonized by AG treatment. Adenine insignificantly reduced the mechanical nociceptive threshold by 15%. The results of the tests for neuromuscular coordination were inconclusive. In conclusion, adenine-induced CRF caused motor and behavioral alterations, and these were significantly mitigated by administration of AG.
Inhibition of adipocyte differentiation and adipogenesis by the traditional Chinese herb Sibiraea angustata
Authors: Xiao Lei , Zhang Jun , Li Hongxing , Liu Jin , He Lan , Zhang Junjie , Zhai Yonggong
Published: April 2011

Obesity has become a major health concern due to its strong association with the metabolic syndrome. Inhibition of adipocyte differentiation represents a key strategy to inhibit obesity. Sibiraea angustata (SA), a traditional Chinese herb, has a wide range of pharmacological effects, such as improving digestive functions. Here, we report a novel antiadipogenic effect of SA. By using the SA water extract (SAW), SA acetic ether extract (SAA) and the 3T3-L1 model of adipocyte differentiation and adipogenesis, we showed that both SAW and SAA impaired the proliferation and adipo-differentiation of 3T3-L1 in a dose- and time-dependent manner. At the molecular level, treatment of 3T3-L1 cells with SAW or SAA inhibited the expression of the key adipocyte differentiation regulator CCAAT enhancer binding protein β (C/EBP β), as well as peroxisome proliferator activated receptor γ, adipocyte protein-2, lipoprotein lipase and glucose transporter 4. Cell cycle analysis showed that both SAW and SAA blocked cell cycle at the G1–S transition phase, causing cells to remain in the preadipocyte state. The expression of CyclinA and cyclin-dependent kinase 2 was also inhibited by SAW and SAA. Treatment with SAW also prevented the localization of C/EBP β to the centromeres. Taken together, our results show that SA has a potent antiadipogenic effect in 3T3-L1 cells due to the inhibition of adipocyte differentiation and adipogenesis. We propose that SA may be used as a safe and effective neutraceutical to manage obesity.
Genetically modified human umbilical cord blood cells expressing vascular endothelial growth factor and fibroblast growth factor 2 differentiate into glial cells after transplantation into amyotrophic lateral sclerosis transgenic mice
Authors: Rizvanov Albert A , Guseva Daria S , Salafutdinov Ilnur I , Kudryashova Nezhdana V , Bashirov Farid V , Kiyasov Andrey P , Yalvaç Mehmet E , Gazizov Ilnaz M , Kaligin Maxim S , Sahin Fikrettin , Mukhamedyarov Marat A , Palotás András , Islamov Rustem R
Published: April 2011

Current therapy of a number of neuropsychiatric maladies has only symptomatic modality. Effective treatment of these neuro-degenerative diseases, including amyotrophic lateral sclerosis (ALS), may benefit from combined gene/stem-cell approaches. In this report, mononuclear fraction of human umbilical cord blood cells (hUCBCs) were transfected by electroporation with dual plasmid constructs, simultaneously expressing vascular endothelial growth factor 165 (VEGF165) and human fibroblast growth factor 2 (FGF2) (pBud-VEGF-FGF2). These genetically modified hUCBCs were injected retro-orbitally into presymptomatic ALS transgenic animal models (G93A mice). Lumbar spinal cords of rodents were processed for immunofluoresent staining with antibodies against human nuclear antigen (HNA), oligodendrocyte-specific protein, S100, iba1, neuronal β3-tubulin and CD34. Co-localization of HNA and S100 was found in the spinal cord of mice after transplantation of genetically modified hUCBCs over-expressing VEGF-FGF2. Double staining in control animals treated with unmodified hUCBCs, however, revealed HNA+ cells expressing iba1 and CD34. Neuron-specific β3-tubulin or oligodendrocyte-specific protein were not expressed in hUCBCs in either control or experimental mice. These results demonstrate that genetically naïve hUCBCs may differentiate into endothelial (CD34+) and microglial (iba1+) cells; however when over-expressing VEGF-FGF2, hUCBCs transform into astrocytes (S100+). Autocrine regulation of VEGF and FGF2 on hUCBCs, signal molecules from dying motor neurons in spinal cord, as well as self-differentiating potential may provide a unique microenvironment for the transformation of hUCBCs into astrocytes that eventually serve as a source of growth factors to enhance the survive potential of surrounding cells in the diseased regions.
Thymoquinone inhibits matrix metalloproteinase expression in rabbit chondrocytes and cartilage in experimental osteoarthritis
Authors: Chen Wei-Ping , Tang Jing-Li , Bao Jia-Peng , Wu Li-Dong
Published: April 2011

Thymoquinone (TQ) is the main constituent of Nigella sativa oil, which has been traditionally used against arthritis in the Middle East. In this study, we investigated the effect of TQ against matrix metalloproteinase (MMP) expression in both rabbit chondrocytes and animal mode of osteoarthritis (OA) induced by anterior cruciate ligament transection and tested whether or not nuclear factor kappa B (NF- κB) was involved in this process. TQ down-regulated MMP-1, MMP-3 and MMP-13 expression and up-regulated tissue inhibitors of metalloproteinase-1 expression as assessed by quantitative realtime polymerase chain reaction. In addition, NF- κB p65 protein level as well as its translocation induced by interleukin-1 β were inhibited by TQ. Our findings suggest the potential of TQ in the treatment of OA.
Preservation of the cardiac function in infarcted rat hearts by the transplantation of adipose-derived stem cells with injectable fibrin scaffolds
Authors: Zhang Xuelian , Wang Haibin , Ma Xiang , Adila Azhati , Wang Baozhu , Liu Fen , Chen Bangdang , Wang Changyong , Ma Yitong
Published: April 2011

Cell-based therapy can improve cardiac function but is limited by the low cell retention and survival within ischemic tissues. Injectable cardiac tissue engineering aims to support cell-based therapies and enhance their efficacy for cardiac diseases. So far, no research has been devoted to studying the usefulness of the combination of fibrin glue (as scaffold) and adipose-derived stem cells (ADSCs) to treat myocardial infarction. In our study, the rat ADSCs were isolated from subcutaneous adipose tissues. The surface phenotype of these cells was analyzed by flow cytometry. The fibrin glue was then co-injected with ADSCs into the left ventricular wall of rat infarction models. The structure and functional consequences of transplantation were determined by detailed histological analysis and echocardiography. Most cultured ADSCs expressed CD105 and CD90, and were negative for CD34 and CD45. After injection, both the 24-h cell retention and four-week graft size were significantly higher and larger in the Fibrin + ADSCs group than those of the ADSCs group alone ( P < 0.01). The heart function improved significantly in the Fibrin + ADSCs group compared with that of the ADSCs group four weeks after transplantation ( P < 0.01). In addition, the arteriole densities within the infarcted area improved significantly in the Fibrin + ADSCs group compared with those in the ADSCs group four weeks after transplantation ( P < 0.01). In conclusion, the ADSCs with the fibrin glue has the therapeutic potential to improve the function of infarcted hearts. The method of in situ injectable tissue engineering combining fibrin glue with ADSCs is promising clinically.
Hepatitis B x-interacting protein induces HepG2 cell proliferation through activation of the phosphatidylinositol 3-kinase/Akt pathway
Authors: Wang Feng-ze , Fei Hong-rong , Lian Li-hui , Wang Jian-mei , Qiu Yu-yu
Published: April 2011

Hepatitis B x-interacting protein (HBXIP), a co-factor of survivin, was originally identified by its binding with the C-terminus of the hepatitis B virus x protein (HBx). We have recently shown that HBXIP promotes the growth of both normal liver cells and hepatoma cells in vitro, but the molecular mechanisms of this have not been documented. In this study, we investigated the potential effects of HBXIP on the proliferation of HepG2 cells and the intracellular signaling pathway mediating these changes. Over-expression of the HBXIP gene promoted the proliferation of HepG2 cells, as shown by the MTT assay. We also showed that HBXIP induced cellular accumulation in the S phase concomitantly with up-regulation of cyclinD1 and down-regulation of p21 and p53 levels. Moreover, HBXIP over-expression cells showed activation of the PI3K/Akt pathway; this activation was accompanied by an increase in phosphorylation of glycogen synthase kinase 3 β. LY294002, a specific inhibitor of PI3K, blocked HBXIP-stimulated Akt phosphorylation and suppressed the cell cycle promotion induced by HBXIP in HepG2 cells. The increase in cyclinD1 protein levels induced by HBXIP was inhibited when cells were incubated with LY294002. In conclusion, our data suggest that the proliferation of HepG2 cells promoted by HBXIP is associated with activation of the PI3K/Akt signaling pathway.
Mutation in fucose synthesis gene ofKlebsiella pneumoniaeaffects capsule composition and virulence in mice
Authors: Pan Po-Chang , Chen Hui-Wen , Wu Po-Kuan , Wu Yu-Yang , Lin Chun-Hung , Wu June H
Published: April 2011

The emerging pathogenicity of Klebsiella pneumoniae (KP) is evident by the increasing number of clinical cases of liver abscess (LA) due to KP infection. A unique property of KP is its thick mucoid capsule. The bacterial capsule has been found to contain fucose in KP strains causing LA but not in those causing urinary tract infections. The products of the gmd and wcaG genes are responsible for converting mannose to fucose in KP. A KP strain, KpL1, which is known to have a high death rate in infected mice, was mutated by inserting an apramycin-resistance gene into the gmd. The mutant expressed genes upstream and downstream of gmd, but not gmd itself, as determined by reverse transcriptase polymerase chain reaction. The DNA mapping confirmed the disruption of the gmd gene. This mutant decreased its ability to kill infected mice and showed decreased virulence in infected HepG2 cells. Compared with wild-type KpL1, the gmd mutant lost fucose in capsular polysaccharides, increased biofilm formation and interacted more readily with macrophages. The mutant displayed morphological changes with long filament forms and less uniform sizes. The mutation also converted the serotype from K1 of wild-type to K2 and weak K3. The results indicate that disruption of the fucose synthesis gene affected the pathophysiology of this bacterium and may be related to the virulence of this KpL1 strain.
CD44 mediates oligosaccharides of hyaluronan-induced proliferation, tube formation and signal transduction in endothelial cells
Authors: Wang Ying Zhi , Cao Man Lin , Liu Yi Wen , He Yi Qing , Yang Cui Xia , Gao Feng
Published: April 2011

Oligosaccharides of hyaluronan (o-HA) can induce angiogenesis and the growth and tube formation of vascular endothelial cells (ECs) in particular. As the major o-HA receptor, CD44 has been implicated in EC function, but its role in mediating o-HA-induced EC proliferation and tube formation remains unclear. In this study, we investigated the role of CD44 in o-HA-induced proliferation and tube formation of human umbilical vein endothelial cells (HUVECs) and explored the molecular mechanisms underlying the angiogenesis process. A CD44 siRNA was delivered into HUVECs by electroporation and o-HA-induced proliferation and tube formation capacity of CD44-silenced or control HUVECs were assessed by methylthiazolyldiphenyl-tetrazolium bromide (MTT) and Matrigel assays. Furthermore, the changes in Src, focal adhesion kinase (FAK) and extracellular signal-regulated kinase1 and 2 (ERK1/2) phosphorylation, as well as the expression of c-jun and c-fos were examined by Western blot and realtime-polymerase chain reaction assays. Our results demonstrated that 10 μg/mL o-HA obviously induced the proliferation and tube formation in HUVECs, and stimulated the phosphorylation of Src, FAK and ERK1/2 and upregulation of c-jun and c-fos, which could be inhibited by CD44 silencing. Altogether our data suggest that CD44 functions to initiate tyrosine phosphorylation of Src, FAK and ERK1/2, and upregulates the expression of c-jun and c-fos, thus mediating o-HA-induced proliferation and tube formation in HUVECs.
The activation of leptin-mediated survivin is limited by the inducible suppressor SOCS-3 in MCF-7 cells
Authors: Palianopoulou Maria , Papanikolaou Vassilis , Stefanou Nikolaos , Tsezou Aspasia
Published: April 2011

Although leptin has been found to be implicated in obesity-related breast carcinogenesis in postmenopausal women, the molecular mechanisms involved are yet to be defined. Recently, the antiapoptotic gene survivin has been recognized as a target gene for leptin in breast cancer. The aim of this study was to investigate the effect of leptin on the expression of survivin and on the transcriptional activity of its promoter in MCF-7 breast cancer cells. We also studied the potential involvement of SOCS-3 (a negative regulator of leptin's main signaling pathway JAK2/STAT3) in the expression of leptin-mediated survivin. Our results showed a significant increase in the mRNA (dose-dependent increase of 40–70%) and protein expression levels of survivin 24 h post-leptin treatment, which was followed by a significant decrease at 48 and 72 h (of 60–70%). In accordance, a chromatin immunoprecipitation assay revealed an initial strong binding of STAT3 to the survivin promoter, which was no longer detected after 24 h. Myc/mad/max network proteins and histone H3 acetylation status were not found to contribute to the expression of leptin-mediated survivin. Furthermore, a protein immunoprecipitation assay detected an enhanced SOCS-3 binding to the long isoform of leptin's receptor (Ob-Rb) 48 and 72 h after leptin administration, thus conferring inhibition to leptin signaling. In conclusion, our findings suggest, for the first time to our knowledge, that the effect of leptin on the antiapoptotic gene survivin is limited by the inhibitory role of SOCS-3 in the leptin-activated JAK2/STAT3 signaling pathway in MCF-7 breast cancer cells.
The burden of obesity on infectious disease
Authors: Karlsson Erik A , Beck Melinda A
Published: April 2011

The world is now experiencing an epidemic of obesity. Although the effects of obesity on the development of metabolic and cardiovascular problems are well studied, much less is known about the impact of obesity on immune function and infectious disease. Studies in obese humans and with obese animal models have repeatedly demonstrated impaired immune function, including decreased cytokine production, decreased response to antigen/mitogen stimulation, reduced macrophage and dendritic cell function, and natural killer cell impairment. Recent studies have demonstrated that the impaired immune response in the obese host leads to increased susceptibility to infection with a number of different pathogens such as community-acquired tuberculosis, influenza, Mycobacterium tuberculosis, coxsackievirus, Helicobacter pylori and encephalomyocarditis virus. While no specific mechanism has been defined for the decreased immune response to infectious disease in the obese host, several obesity-associated changes such as excessive inflammation, altered adipokine signaling, metabolic changes and even epigenetic regulation could affect the immune response. This review will discuss what is currently known about the relationship between obesity and infectious disease.
Targeting transforming growth factor βRII expression inhibits the activation of hepatic stellate cells and reduces collagen synthesis
Authors: Fu Rongquan , Wu Jinguo , Ding Jiguang , Sheng Jifang , Hong Liang , Sun Qingfeng , Fang Hui , Xiang Dairong
Published: April 2011

Abnormal production of extracellular matrix (ECM) components significantly contributes to the development of liver fibrosis. This study aimed at examining the effects of short-hairpin RNA (shRNA)-mediated transient knockdown of transforming growth factor βRII (TGF βRII) expression on the proliferation and activation of hepatic stellate cells (HSCs) and synthesis of fibrogenic ECM components in HSC cells. Three different shRNA-expressing plasmids were constructed for the expression of shRNA-(a, b, c) targeting to the rat TGF βRII mRNA beginning at nucleotide position 339, 444 and 528 and they were transfected into a rat stellate cell line, HSC-T6 cells, respectively. The levels of TGF βRII, α-smooth muscle actin ( α-SMA), and type I and III collagen expressions were characterized by reverse transcription polymerase chain reaction and Western blot assays. The concentrations of hyaluronic acid (HA) and type IV collagen in the supernatants of cultured cells were measured by enzyme-linked immunosorbent assay. Transfection with the TGF βRII-specific shRNAs resulted in varying levels of inhibition in the expression of TGF βRII in HSC-T6 cells, and transfection with the potent shRNA-c inhibited the expression of TGF βRII in a dose-dependent manner. Knockdown of TGF βRII expression significantly reduced the levels of α-SMA, type I, III and IV collagen, and HA expression in HSC-T6 cells ( P < 0.01). In conclusion, our data indicated that knockdown of TGF βRII expression inhibited the activation of HSCs and the production of fibrogenic ECM components in HSC-T6 cells. Therefore, our findings support the notion that TGF βRII is an important factor of the pathogenic process of liver fibrosis.
The role of magnetic forces in biology and medicine
Authors: Roth Bradley J
Published: April 2011

The Lorentz force (the force acting on currents in a magnetic field) plays an increasingly larger role in techniques to image current and conductivity. This review will summarize several applications involving the Lorentz force, including (1) magneto-acoustic imaging of current; (2) ‘Hall effect’ imaging; (3) ultrasonically-induced Lorentz force imaging of conductivity; (4) magneto-acoustic tomography with magnetic induction; and (5) Lorentz force imaging of action currents using magnetic resonance imaging.
Hierarchical architecture influences calcium dynamics in engineered cardiac muscle
Authors: Pong Terrence , Adams William J , Bray Mark-Anthony , Feinberg Adam W , Sheehy Sean P , Werdich Andreas A , Parker Kevin Kit
Published: April 2011

Changes in myocyte cell shape and tissue structure are concurrent with changes in electromechanical function in both the developing and diseased heart. While the anisotropic architecture of cardiac tissue is known to influence the propagation of the action potential, the influence of tissue architecture and its potential role in regulating excitation–contraction coupling (ECC) are less well defined. We hypothesized that changes in the shape and the orientation of cardiac myocytes induced by spatial arrangement of the extracellular matrix (ECM) affects ECC. To test this hypothesis, we isolated and cultured neonatal rat ventricular cardiac myocytes on various micropatterns of fibronectin where they self-organized into tissues with varying degrees of anisotropy. We then measured the morphological features of these engineered myocardial tissues across several hierarchical dimensions by measuring cellular aspect ratio, myocyte area, nuclear density and the degree of cytoskeletal F-actin alignment. We found that when compared with isotropic tissues, anisotropic tissues have increased cellular aspect ratios, increased nuclear densities, decreased myocyte cell areas and smaller variances in actin alignment. To understand how tissue architecture influences cardiac function, we studied the role of anisotropy on intracellular calcium ([Ca2+]i) dynamics by characterizing the [Ca2+]i–frequency relationship of electrically paced tissues. When compared with isotropic tissues, anisotropic tissues displayed significant differences in [Ca2+]i transients, decreased diastolic baseline [Ca2+]i levels and greater [Ca2+]i influx per cardiac cycle. These results suggest that ECM cues influence tissue structure at cellular and subcellular levels and regulate ECC.
Effect of Perilla frutescens var. acuta Kudo and rosmarinic acid on allergic inflammatory reactions
Authors: Oh Hyun-A , Park Cheung-Seog , Ahn Hyun-Jong , Park Yong Seek , Kim Hyung-Min
Published: April 2011

Allergy is characterized by an overreaction of the immune system. Perilla frutescens leaf extract has been reported to exhibit antiallergic inflammatory activity. To investigate precisely the effect and mechanism of 30% ethanol extract powder of P. frutescens var. acuta Kudo (EPPF) and rosmarinic acid (RA), a component of EPPF in allergic rhinitis and rhinoconjunctivitis, the antiallergic effects of EPPF and RA were analyzed using in vivo and in vitro models. Cytokine production was analyzed by means of an enzyme-linked immunosorbent assay. Cytokine expression was analyzed via reverse transcription-polymerase chain reaction and Western blotting. Transcription factor and caspase-1 activity were analyzed by a luciferase assay and caspase-1 assay, respectively. The number of nasal, ear and eye rubs after an ovalbumin (OVA) challenge in OVA-sensitized mice was significantly higher than that in OVA-unsensitized mice. Increased number of rubs was inhibited by administration of EPPF or RA. Increased levels of IgE in the serum, spleen and nasal mucosa of OVA-sensitized mice were reduced by EPPF or RA administration. The histamine level was also reduced by EPPF or RA administration in the serum of OVA-sensitized mice. Protein levels and mRNA expressions of interleukin (IL)-1 β, IL-6 and tumor necrosis factor- α were inhibited by EPPF or RA administration in the nasal mucosa tissue or spleen of OVA-sensitized mice. In EPPF or RA-administered mice, the mast cell and eosinophil infiltration increase as caused by OVA-sensitization was decreased. In addition, EPPF or RA inhibited both cyclooxygenase-2 protein expression and caspase-1 activity in the same nasal mucosa tissue. In activated human mast cells, nuclear factor-kappa B (NF- κB) /Rel A and caspase-1 activation increased, whereas NF- κB /Rel A and caspase-1 activation was inhibited after a treatment of EPPF or RA. These results indicate that EPPF and RA ameliorate allergic inflammatory reactions such as allergic rhinitis and allergic rhinoconjunctivitis.
Experimental models of Rett syndrome based on Mecp2 dysfunction
Authors: Calfa Gaston , Percy Alan K , Pozzo-Miller Lucas
Published: April 2011

Rett syndrome (RTT) is a neurodevelopmental disorder predominantly occurring in females with an incidence of 1:10,000 births and caused by sporadic mutations in the MECP2 gene, which encodes methyl-CpG-binding protein-2, an epigenetic transcription factor that binds methylated DNA. The clinical hallmarks include a period of apparently normal early development followed by a plateau and then subsequent frank regression. Impaired visual and aural contact often lead to an initial diagnosis of autism. The characterization of experimental models based on the loss-of-function of the mouse Mecp2 gene revealed that subtle changes in the morphology and function of brain cells and synapses have profound consequences on network activities that underlie critical brain functions. Furthermore, these experimental models have been used for successful reversals of RTT-like symptoms by genetic, pharmacological and environmental manipulations, raising hope for novel therapeutic strategies to improve the quality of life of RTT individuals.
Suppression of mast-cell-mediated allergic inflammation byLindera obtusiloba
Authors: Suh Won Mo , Park Seung-Bin , Lee Soyoung , Kim Hui-Hun , Suk Kyoungho , Son Jun-Ho , Kwon Taeg Kyu , Choi Hyun Gyu , Lee Seung-Ho , Kim Sang-Hyun
Published: April 2011

Allergic disease is a consequence of exposure to normally innocuous substances that elicit the activation of mast cells. Mast-cell-mediated allergic response is involved in many diseases such as anaphylaxis, allergic rhinitis, asthma and atopic dermatitis. The discovery of drugs for the treatment of allergic disease is an important subject in human health. In this study, we investigated the effect of Lindera obtusiloba water extract (LOWE) on the mast-cell-mediated allergic inflammation and possible mechanism of action using in vitro and in vivo models. LOWE reduced histamine release from various types of mast cells activated by immunoglobulin E (IgE) or phorbol 12-myristate 13-acetate and calcium ionophore A23187 (PMACI). The inhibitory effect of LOWE on histamine release was mediated by calcium signal. LOWE decreased the PMACI-stimulated gene expression of proinflammatory cytokines such as tumor necrosis factor- α and interleukin-6 in human mast cells. The inhibitory effect of LOWE on the proinflammatory cytokines was nuclear factor (NF)- κB dependent. In addition, LOWE suppressed compound 48/80-induced systemic allergic reaction and serum histamine release in mice and IgE-mediated local allergic reactions. Our results indicate that LOWE inhibits mast-cell-derived allergic inflammation and involvement of calcium, histamine, proinflammatory cytokines and NF- κB in these effects.
Salmonella-mediated gene therapy in experimental colitis in mice
Authors: Palffy Roland , Gardlik Roman , Behuliak Michal , Jani Peter , Balakova Denisa , Kadasi Ludevit , Turna Jan , Celec Peter
Published: April 2011

Bacterial gene therapy – bactofection is a simple and effective method to deliver plasmid DNA into target tissue. We hypothesize that oral in vivo bactofection can be an interesting approach to influence the course of inflammatory bowel diseases. The aim of this study was to prove the effects of antioxidative and anti-inflammatory bactofection in dextran sulfate sodium (DSS)-treated mice. Attenuated bacteria Salmonella Typhimurium SL7207 carrying plasmids with genes encoding Cu-Zn superoxide dismutase and an N-terminal deletion mutant of monocyte chemoattractant protein-1 were prepared. Male Balb/c mice had ad libitum access to 1% DSS solution in drinking water during 10 days (mild model of colitis). The animals were daily fed with 200 Mio bacteria via gastric gavage during the experiment. Fecal consistency, clinical status, food and water intake were monitored. After 10 days samples were taken and markers of oxidative stress and inflammatory cytokine levels were measured. Colonic tissue was scored histologically by a blinded investigator. DSS treatment significantly increased the levels of inflammatory cytokines and malondialdehyde as a marker of lipoperoxidation in the colon. Anti-inflammatory gene therapy improved the total antioxidative capacity. In comparison with the untreated group, bacterial gene therapy lowered the histological colitis score. Salmonella-mediated antioxidative and anti-inflammatory gene therapy alleviated colitis in mice. The effect seems to be mediated by increased antioxidative status. Further studies will show whether recombinant probiotics expressing therapeutic gene might be used for the therapy of inflammatory bowel diseases.
Proteomic analysis of serum proteins in acute ischemic stroke patients treated with acupuncture
Authors: Pan Sanqiang , Zhan Xiangli , Su Xuan , Guo Lei , Lv Laiqing , Su Baogui
Published: April 2011

In the present study, we have investigated the effects of acupuncture on (1) serum protein expression that might have a beneficial effect on stroke patients and (2) the strength of limb muscles in stroke patients. A total of 35 acute ischemic stroke (IS) patients were divided into two groups, one receiving drug treatment alone and the other receiving electroacupuncture (EA) and drug treatment. EA treatment was performed on eight acupuncture points once a day for 10 consecutive days. Serum proteins were detected using a proteomics method based on two-dimensional gel electrophoresis, and the specificity of proteins was confirmed by Western blotting. Changes of limb muscle strength were measured using a modified Medical Research Council grading scale. After EA, SerpinG1 protein expression in serum was down-regulated while the expressions of gelsolin, complement component I, C3, C4B and beta-2-glycoprotein I proteins were up-regulated in patients. The changes of serum protein expression were further confirmed by Western blotting in a majority of the cases. The muscle strength of limbs was increased after EA in 18 patients. EA appears to be effective in regulating differential expression of multiple serum proteins involved in stroke, and also in enhancement of muscle strength recovery in acute IS patients despite an individual variation.
Carbon black nanoparticles enhance bleomycin-induced lung inflammatory and fibrotic changes in mice
Authors: Kamata Hirofumi , Tasaka Sadatomo , Inoue Ken-ichiro , Miyamoto Keisuke , Nakano Yasushi , Shinoda Hiromi , Kimizuka Yoshifumi , Fujiwara Hiroshi , Ishii Makoto , Hasegawa Naoki , Takamiya Rina , Fujishima Seitaro , Takano Hirohisa , Ishizaka Akitoshi
Published: April 2011

With the recent increasing use of nanoparticles, there is concern that they may become an environmental risk factor as airborne particles. However, the impact of these particles on susceptible subjects with predisposing lung disease have not been sufficiently elucidated. In the present study, we investigated the effects of nanoparticles on pulmonary inflammatory and fibrotic changes induced by intratracheal bleomycin (BLM) challenge in mice. Mice were intratracheally administered either vehicle, 14-nm carbon black nanoparticles (CBNPs), BLM or BLM plus CBNP. First, we assessed lung collagen content, lung compliance and fibrotic changes in histopathology on day 21 after instillation. Then, to elucidate how CBNP contributes to the development of BLM-induced fibrosis, we collected bronchoalveolar lavage (BAL) fluid on days 2, 7, 14 and 21 and determined the total and differential cell counts and concentrations of two proinflammatory cytokines (keratinocyte chemoattractant [KC] and interleukin [IL]-6) and two fibrogenic mediators (CC chemokine ligand 2 [CCL2] and transforming growth factor- β1 [TGF- β1]). Expression of nitrotyrosine, an indicator of oxidant injury, was also evaluated on days 7 and 21. CBNP, when combined with BLM, significantly enhanced BLM-induced increase in lung collagen content, decrease in lung compliance, and fibrotic changes in histopathology. CBNP significantly augmented BLM-induced increase in the numbers of inflammatory cells in BAL fluid on days 2 and 7 and levels of KC and IL-6 on day 2. In addition, CBNP administered in combination with BLM significantly elevated the levels of CCL2 on days 2, 7 and 14, and TGF- β1 on day 14 in BAL fluid as compared with BLM alone. Nitrotyrosine expression was also increased by BLM plus CBNP compared with BLM alone. In contrast, CBNP did not exert any significant effect on these parameters by itself. These results indicate that CBNP can exaggerate BLM-induced inflammatory and fibrotic changes in the lung, suggesting the potential impact of nanoparticles on lung inflammation and fibrosis.
Increased antiprotease activity of the SERPINB3 polymorphic variant SCCA-PD
Authors: Turato Cristian , Biasiolo Alessandra , Pengo Paolo , Frecer Vladimir , Quarta Santina , Fasolato Silvano , Ruvoletto Mariagrazia , Beneduce Luca , Zuin Jessica , Fassina Giorgio , Gatta Angelo , Pontisso Patrizia
Published: April 2011

SERPINB3 has been found in chronic liver damage and hepatocellular carcinoma, but not in normal liver. By direct mRNA sequencing, a new SERPINB3 polymorphism (SCCA-PD) has been identified, presenting the substitution Gly351Ala in the reactive center loop of the protein. The prevalence of the SCCA-PD isoform has been found to be significantly higher in patients with cirrhosis than in patients with chronic liver disease and in normal subjects. The aim of this study was to investigate the biological and functional activity of SERPINB3 isoforms using in vitro models. HepG2 and Huh7 cells lines were transfected with plasmid vectors containing wild-type SERPINB3 or its polymorphic variant SCCA-PD and their expression at transcriptional and protein level was determined. To assess the functional activity, both recombinant proteins were produced and kinetic analysis was carried out using papain and cathepsin-L as target proteases. In addition, the inhibition of JNK kinase activity by SERPINB3 isoforms was assessed. The crystal structure of wild-type SERPINB3 at 2.7 Å resolution was used for preparation of refined 3D models of the two isoforms. The results showed that transcriptional activity and protein expression of the two isoforms were similar in both transfected cell lines. Both SERPINB3 preparations exerted a dose-dependent protease inhibitory activity, but the effect of SCCA-PD was higher than that of the wild-type isoform. This result was supported by 3D modelling, where increased hydrophobic profile of the SCCA-PD isoform, introduced by the G351A mutation, was detected. In addition, at high protein concentration, SCCA-PD revealed a 16% higher inhibitory effect on c-Jun phosphorylation by JNK1, compared with wild-type SERPINB3. In conclusion, the single amino acid substitution in the SERPINB3 reactive site loop improves the functional activity of SCCA-PD isoform. This different antiprotease activity might favor disease progression in patients carrying this polymorphism.
Predictors of condom errors among sex workers in Armenia
Authors: Lang D L , Salazar L F , DiClemente R J , Markosyan K , Darbinyan N
Published: April 2011

This cross-sectional study identified the prevalence and correlates of condom-use errors among female sex workers (FSWs) in Armenia. One hundred and seventeen street-based FSWs aged 20–52 years completed an interviewer-administered questionnaire. Condom-use errors were reported by 78.0% of participants. Number of clients, higher frequency of condom application on clients by FSWs, greater perceived barriers to condom use, elevated depressive symptomatology and having sex while drinking alcohol were significantly associated with higher number of condom-use errors. History of sexually transmitted infections (STIs) was marginally significant while consistent condom use was not significant in the final model. The multiple regression model accounted for 32.5% of the variance in condom-use errors. Condom-use errors are prevalent in this population, thus attenuating the intended protective effects of condoms. Interventions with FSWs in Armenia should specifically address the factors identified in this study toward the goal of reducing condom errors and ultimately preventing acquisition of STIs including HIV.
Attitudes and first heterosexual experiences among Indians and Pakistanis in Britain: evidence from a national probability survey
Authors: Griffiths C , Johnson A M , Fenton K A , Erens B , Hart G J , Wellings K , Mercer C H
Published: April 2011

We compare attitudes, experiences of learning about sex and first intercourse among Indians ( n = 393) and Pakistanis ( n = 365) using a probability survey of Britain's general population aged 16–44 years conducted during 1999–2001 ( n = 12,110). Higher proportions of Pakistanis (64.6%) and Indians (28.1%) reported religion as ‘very important’ versus 6.2% of other ethnicities. Pakistanis were more conservative in their attitudes, e.g. reporting premarital sex as wrong (adjusted odds ratios [AORs] for sociodemographic differences, 4.71 [men] and 6.59 [women]). Pakistanis were more likely to be married at first sex (AORs 6.2 [men] and 9.53 [women]), yet men were more likely than women to be in non-marital relationships at this time (69.4% versus 25.2%). Pakistani men and women and Indian women were more likely to report not using reliable contraception at first sex relative to others (AORs 2.33, 3.16 and 1.90, respectively). Pakistani and Indian women were more likely than others to report school lessons as their main source of sex education (AORs 2.23 and 1.77) and not discussing sex with their parents during adolescence (AORs 2.04 and 2.62). These unique data have implications for ensuring that sex and relationship education and health promotion messages are appropriately planned, targeted and delivered to benefit Pakistanis and Indians.
Effect of antiretroviral therapy on HIV-1 genetic evolution during acute infection
Authors: Chamberland A , Sylla M , Boulassel M R , Baril J-G , Côté P , Thomas R , Trottier B , Rouleau D , Routy J-P , Tremblay C
Published: April 2011

The rapid evolution of HIV-1 is a major obstacle to viral eradication. Early antiretroviral therapy (ART) during primary HIV-1 infection could limit viral diversity. Eighteen patients recently infected with HIV-1 were selected. Nine initiated ART soon after enrolment and nine remained untreated. Replication-competent (RC) viruses were quantified at baseline and after one year of follow-up. Viral diversity in the C2V5 envelope region was evaluated from plasma, peripheral blood mononuclear cells (PBMCs), and cell culture at both time points. The amount of RC virus in the treated group declined (median −5.42 infectious units per million [IUPM]) while it remained stable or increased in the untreated group (median +0.87 IUPM). At one year post infection, we observed a significant increase in diversity for the C2V5 (+0.150%) region, specifically in the hypervariable loops V4 (+0.73%) and V5 (+0.77%), in the untreated group. More importantly, viral diversity did not significantly increase in treated individuals during the first year post infection. Genetic diversity during primary infection remains low through the first year of infection. Early treatment could contribute to a decrease in RC viruses from PBMCs and to limitation of viral diversification in the viral reservoir. These findings may have relevance for the rational design of specific immunotherapeutic strategies.
Patterns of HIV testing at a London teaching hospital between 2004 and 2007
Authors: Madge S , Smith C , Evans A , Clewley G , Johnson M A , Geretti A M
Published: April 2011

To observe trends in testing for HIV between 2004 and 2007 in a London hospital and to observe the seroprevalence of HIV within subgroups. Tests were grouped according to source and reason for testing; 58,720 tests were considered (HIV-1 seroprevalence = 0.9%). The majority (75.4%) of tests were performed as part of routine and opt-out protocols including antenatal and genitourinary screening. Among people specifically seeking HIV testing, the HIV seroprevalence was 3.5%. Medical specialties performed fewer tests but a high HIV seroprevalence was observed, including infectious disease (seroprevalence 4.4%) and other medical specialties (seroprevalence 3.4%). A small number of specialties performed few HIV tests. HIV testing was cost-effective in virtually all settings. This study suggests that more HIV tests could be performed, for example, in acute medicine, and training might increase the number of tests offered in some settings. The most effective way of increasing testing appears to be opt-out testing.
Are we forgetting the children? Testing the children of HIV-positive parents
Authors: Whitfield C A , Kingston M , Fothergill H , Thng C
Published: April 2011

Undiagnosed HIV infection in children carries significant risks and national guidelines recommend that children of HIV-positive parents be tested. We reviewed the testing of HIV-positive patients' children in those new to our service from April 2008 to December 2009. Of the 86 women, 95% were asked about children. Of the 243 men, 53 were heterosexual or bisexual, and 190 identified themselves as men who have sex with men (MSM). Parental status was determined in 37 (70%) of heterosexual and bisexual men and in 14 (17%) of 84 MSM with a history of previous female partners. All untested children apart from one lived in their country of origin. In MSM, recording of parental status was uncommon. Following this audit, clinic proformas were changed to document the parental status of men of all sexual orientations, and the HIV status of all children.
Prevalence of genital chlamydial infection among a community sample of young international backpackers in Sydney, Australia
Authors: Davies S C , Karagiannis T , Headon V , Wiig R , Duffy J
Published: April 2011

We conducted a chlamydia prevalence study from January to June 2009 among a community sample of young international backpackers by recruiting at hostels in Sydney, Australia. Participants completed a questionnaire; men provided a urine sample and women provided a self-collected vaginal swab, which were tested for Chlamydia trachomatis DNA by strand displacement amplification. We recruited 225 men (median age 24 years) and 207 women (median age 23 years). Most (87%) of the travellers came from Europe. A new sexual partner during travel was reported by 67%, and 51% had more than one new sexual partner. Of those reporting a new sexual partner, 40% always used condoms. Prevalence of chlamydia was 3.5% (3.1% in men, 3.9% in women). Previous testing for chlamydia was reported by 40%. Drinking alcohol at hazardous levels was reported by 58% of men and 29% of women. Despite the reporting of new sexual partners and inconsistent condom use, the prevalence of chlamydia in these backpackers was not higher than that found in more general populations, and may relate to good health-care seeking behaviour. Young travellers need education about sexual risks and promotion of condom use prior to travel, and access to public sexual health services.
Evaluation of the use of a social networking site in sexual health care
Authors: Hedge K C , Donald C P
Published: April 2011

The Myspace project was launched in 2008 to develop a novel approach to engage with younger sexual health service users. The aim was to use a social networking site as a patient forum, to gain feedback on how to improve services and to make sexual health information accessible to a larger audience, many of whom might not be current service users and thereby to encourage attendances from more ‘hard to reach’ groups. In this report, we explore the effectiveness and specific challenges of the project and in particular, the difficulty in achieving involvement in the discussion forum. We discuss the future of the project and make recommendations to other services considering this approach.