bluuebunny/crossref_metadata_2025 huggingface dataset:
Therapeutic potential of bacteriophage in treating Klebsiella pneumoniae B5055-mediated lobar pneumonia in mice
Authors: Chhibber Sanjay , Kaur Sandeep , Kumari Seema
Published: November 2008

Klebsiella pneumoniaecauses infections in humans especially in immunocompromised patients. About 80 % of nosocomial infections caused byK. pneumoniaeare due to multidrug-resistant strains. The emergence of antibiotic-resistant bacterial strains necessitates the exploration of alternative antibacterial therapies, which led our group to study the ability of bacterial viruses (known as bacteriophages or simply phages) to treat mice challenged withK. pneumoniae. Phage SS specific forK. pneumoniaeB5055 was isolated and characterized, and its potential as a therapeutic agent was evaluated in an experimental model ofK. pneumoniae-mediated lobar pneumonia in mice. Mice were challenged by intranasal (i.n.) inoculation with bacteria (108 c.f.u. ml−1). A single intraperitoneal injection of 1010 p.f.u. ml−1phage administered immediately after i.n. challenge was sufficient to rescue 100 % of animals fromK. pneumoniae-mediated respiratory infections. Administration of the phage preparation 3 h prior to i.n. bacterial challenge provided significant protection in infected mice, while even 6 h delay of phage administration after the induction of infection rendered the phage treatment ineffective. The results of this study therefore suggest that the timing of starting the phage therapy after initiation of infection significantly contributes towards the success of the treatment.
Persistent Candida parapsilosis funguria associated with an indwelling urinary tract stent for more than 7 years
Authors: Ruan Sheng-Yuan , Chien Jung-Yien , Hsueh Po-Ren
Published: November 2008

Candiduria is an increasingly common condition, and the lack of effective antifungal treatment in many cases has raised great concern. We report a case of persistent Candida parapsilosis funguria associated with urinary tract instrumentation. Molecular typing suggested that during a 7 year period the C. parapsilosis isolates were all the same strain. Prolonged antifungal therapy and regular catheter replacement failed to eradicate the funguria, but improved urinary symptoms and pyuria. The antifungal susceptibility pattern did not significantly change during the clinical course despite repeated exposure to fluconazole.
Comparison of virulence factors in Pseudomonas aeruginosa strains isolated from contact lens- and non-contact lens-related keratitis
Authors: Choy Man H. , Stapleton Fiona , Willcox Mark D. P. , Zhu Hua
Published: November 2008

Pseudomonas aeruginosa is one of the common pathogens associated with corneal infection, particularly in contact lens-related keratitis events. The pathogenesis of P. aeruginosa in keratitis is attributed to the production of virulence factors under certain environmental conditions. The aim of this study was to determine differences in the virulence factors of P. aeruginosa isolated from contact lens- and non-contact lens-related keratitis. Associations were assessed between type III secretion toxin-encoding genes, protease profiles, biofilm formation, serotypes and antibiotic-resistance patterns among 27 non-contact lens- and 28 contact lens-related P. aeruginosa keratitis isolates from Australia. Strains with a exoS
+/exoU
− genotype and a type I protease profile predominated in the non-contact lens-related keratitis isolates, whereas the exoS
−/exoU
+ and a type II protease profile was associated with contact lens-related isolates (P<0.05). A strong biofilm formation phenotype was found to be associated with the possession of the exoU gene, and serotypes E, I and C. The exoS gene was strongly associated with serotypes G, A and B, while exoU was associated with serotypes E and C. Six out of fifty-five (11 %) clinical isolates were non-susceptible (intermediate-resistant or resistant) to ofloxacin and moxifloxacin. All resistant isolates were from non-contact lens-related keratitis. The results suggest that P. aeruginosa isolates from different infection origins may have different characteristics. A better understanding of these differences may lead to further development of evidence-based clinical guidelines for the management of keratitis.
Isolation of Bartonella species from rodents in Taiwan including a strain closely related to ‘Bartonella rochalimae’ from Rattus norvegicus
Authors: Lin Jen-Wei , Chen Chun-Yu , Chen Wan-Ching , Chomel Bruno B. , Chang Chao-Chin
Published: November 2008

An increasing number of Bartonella species originally isolated from small mammals have been identified as emerging human pathogens. During an investigation of Bartonella infection in rodent populations carried out in Taiwan in 2006, a total of 58 rodents were tested. It was determined that 10.3 % (6/58) of the animals were Bartonella bacteraemic. After PCR/RFLP analysis, four isolates were identified as Bartonella elizabethae and one isolate as Bartonella tribocorum. However, there was one specific isolate with an unrecognized PCR/RFLP pattern. After further sequence and phylogenetic analyses of the gltA, ftsZ and rpoB genes, and the 16S–23S rRNA intergenic spacer region, the results indicated that this specific isolate from Rattus norvegicus was closely related to human pathogenic ‘Bartonella rochalimae’. Further studies need to be conducted to evaluate whether this rodent species could be a reservoir for ‘B. rochalimae’.
Intra-individual diversity and similarity of salivary and faecal microbiota
Authors: Maukonen Johanna , Mättö Jaana , Suihko Maija-Liisa , Saarela Maria
Published: November 2008

In the present study, polyphasic analysis [cultivation, combined with the fingerprinting of individual isolates, and denaturing gradient gel electrophoresis (DGGE)] was applied to study whether similar features concerning the diversity and temporal stability of selected bacterial groups could be detected intra-individually in two different niches – the oral cavity and the colon – from ten adult volunteers consuming probiotics. The predominant bacterial microbiota, Clostridium coccoides–Eubacterium rectale group and bifidobacterial populations, were generally stable in salivary and faecal samples, with the greater diversity seen in faeces. Furthermore, different species predominated at the two different sites. Lactobacillus group DGGE profiles were unstable, yet the intra-individual profiles from faecal and salivary samples collected at the same time resembled each other. The ingested probiotic product did not affect the stability of the bacterial groups studied. The culture-based analysis showed that most subjects harboured identical indigenous Lactobacillus genotypes in saliva and faeces (Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus paracasei and Lactobacillus plantarum group). Thus, identical indigenous lactobacilli were able to inhabit both ends of the orogastrointestinal tract, whereas the composition of the other bacterial groups studied varied between the two sites.
Intestinal TM7 bacterial phylogenies in active inflammatory bowel disease
Authors: Kuehbacher Tanja , Rehman Ateequr , Lepage Patricia , Hellmig Stephan , Fölsch Ulrich R. , Schreiber Stefan , Ott Stephan J.
Published: November 2008

TM7 is a recently described subgroup of Gram-positive uncultivable bacteria originally found in natural environmental habitats. An association of the TM7 bacterial division with the inflammatory pathogenesis of periodontitis has been previously shown. This study investigated TM7 phylogenies in patients with inflammatory bowel diseases (IBDs). The mucosal microbiota of patients with active Crohn's disease (CD; n=42) and ulcerative colitis (UC; n=31) was compared with that of controls (n=33). TM7 consortia were examined using molecular techniques based on 16S rRNA genes, including clone libraries, sequencing and in situ hybridization. TM7 molecular signatures could be cloned from mucosal samples of both IBD patients and controls, but the composition of the clone libraries differed significantly. Taxonomic analysis of the sequences revealed a higher diversity of TM7 phylotypes in CD (23 different phylotypes) than in UC (10) and non-IBD controls (12). All clone libraries showed a high number of novel sequences (21 for controls, 34 for CD and 29 for UC). A highly atypical base substitution for bacterial 16S rRNA genes associated with antibiotic resistance was detected in almost all sequences from CD (97.3 %) and UC (100 %) patients compared to only 65.1 % in the controls. TM7 bacteria might play an important role in IBD similar to that previously described in oral inflammation. The alterations of TM7 bacteria and the genetically determined antibiotic resistance of TM7 species in IBD could be a relevant part of a more general alteration of bacterial microbiota in IBD as recently found, e.g. as a promoter of inflammation at early stages of disease.
Two Leucine-Rich Repeat Receptor Kinases Mediate Signaling, Linking Cell Wall Biosynthesis and ACC Synthase in Arabidopsis
Authors: Xu Shou-Ling , Rahman Abidur , Baskin Tobias I. , Kieber Joseph J.
Published: November 2008

The plant cell wall is a dynamic structure that changes in response to developmental and environmental cues through poorly understood signaling pathways. We identified two Leu-rich repeat receptor-like kinases in Arabidopsis thaliana that play a role in regulating cell wall function. Mutations in these FEI1 and FEI2 genes (named for the Chinese word for fat) disrupt anisotropic expansion and the synthesis of cell wall polymers and act additively with inhibitors or mutations disrupting cellulose biosynthesis. While FEI1 is an active protein kinase, a kinase-inactive version of FEI1 was able to fully complement the fei1 fei2 mutant. The expansion defect in fei1 fei2 roots was suppressed by inhibition of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase, an enzyme that converts Ado-Met to ACC in ethylene biosynthesis, but not by disruption of the ethylene response pathway. Furthermore, the FEI proteins interact directly with ACC synthase. These results suggest that the FEI proteins define a novel signaling pathway that regulates cell wall function, likely via an ACC-mediated signal.
The accuracy of the FIB‐4 index for the diagnosis of mild fibrosis in chronic hepatitis B
Authors: MALLET V. , DHALLUIN‐VENIER V. , ROUSSIN C. , BOURLIERE M. , PETTINELLI M. E. , GIRY C. , VALLET‐PICHARD A. , FONTAINE H. , POL S.
Published: November 2008

Background  The Fib‐4 index is a simple and inexpensive biomarker to delineate liver fibrosis in chronic hepatitis C.

Aim  To assess the accuracy of the FIB‐4 index in chronic hepatitis B.

Methods  We compared the FIB‐4 index with 138 synchronous liver biopsies and with 372 synchronous FibroTest performed either in France or in an endemic area (Mayotte, an overseas collectivity of France).

Results  The FIB‐4 index allowed the correct identification of patients with nil‐to‐moderate fibrosis with an area under the receiving operating characteristic curve of 0.81 (P < 0.001), increasing as a function of the length of the liver biopsy (up to 0.94 for liver biopsies ≥20 mm). A cut‐off value ≤1.45 differentiated moderate fibrosis from severe fibrosis with a negative predictive value of 86%, a sensitivity of 71.1% and a specificity of 73.1%. Beyond 1.45, the FIB‐4 index was not informative. The FIB‐4 index was more precise than the AST‐to‐platelet ratio index and correlated with the FibroTest in 89% of the cases (κ = 0.27, P < 0.001) to exclude severe fibrosis.

Conclusion  The FIB‐4 index is a simple, accurate and inexpensive method to exclude significant liver fibrosis in chronic hepatitis B, a major advantage in HBV‐endemic developing countries.
Symptoms in patients on long‐term proton pump inhibitors: prevalence and predictors
Authors: RAGHUNATH A. S. , HUNGIN A. P. S. , MASON J. , JACKSON W.
Published: November 2008

Background  Symptom control in primary care patients on long‐term proton pump inhibitor (PPI) treatment is poorly understood.

Aim  To explore associations between symptom control and demographics, lifestyle, PPI use, diagnosis and Helicobacter pylori status.

Methods  A cross‐sectional survey (n = 726) using note reviews, questionnaires and carbon‐13 urea breath testing. Determinants of symptom control [Leeds Dyspepsia Questionnaire (LDQ), Carlsson and Dent Reflux Questionnaire (CDRQ), health‐related quality‐of‐life measures (EuroQoL: EQ‐5D and EQ‐VAS)] were explored using stepwise linear regression.

Results  Moderate or severe dyspepsia symptoms occurred in 61% of subjects (LDQ) and reflux symptoms in 59% (CDRQ). Age, gender, smoking and body mass index had little or no influence upon symptom control or PPI use. Average symptom scores and PPI use were lower in patients with non‐ulcer dyspepsia and gastro‐protection than gastro‐oesophageal reflux disease (GERD) and uninvestigated dyspepsia. H. pylori infection was associated with lower reflux symptom scores only in patients with GERD and uninvestigated dyspepsia. EQ‐5D was not able to discriminate between diagnostic groups, although the EQ‐VAS performed well.

Conclusions  A majority of patients suffered ongoing moderate or severe symptoms. GERD and uninvestigated dyspepsia were associated with poorer long‐term symptom control; H. pylori appeared to have a protective effect on reflux symptoms in these patients.
Quantitative colonoscopic evaluation of relative efficiencies of an immunochemical faecal occult blood test and a sensitive guaiac test for detecting significant colorectal neoplasms
Authors: ROZEN P. , LEVI Z. , HAZAZI R. , WAKED A. , VILKIN A. , MAOZ E. , BIRKENFELD S. , NIV Y.
Published: November 2008

Background  The guaiac faecal occult blood test (G‐FOBT), HemoccultSENSA, is sensitive for significant neoplasms [colorectal cancer (CRC), advanced adenomatous polyps (AAP)], but faulted by non‐specificity for human haemoglobin (Hb). Quantified, Hb‐ specific, immunochemical faecal occult blood tests (I‐FOBT) are now used.

Aims  To (i) compare I‐FOBT and G‐FOBT efficacy in identifying significant neoplasms and colonoscopy needs for positive tests and (ii) examine number of I‐FOBTs needed and test threshold to use for equivalent or better sensitivity than G‐FOBT and fewest colonoscopies for positive tests.

Methods  Three daily G‐FOBTs and I‐FOBTs were collected and analysed in 330 patients scheduled for colonoscopy.

Results  Colonoscopy found significant neoplasms in 32 patients, 6 CRC, 26 AAP. G‐FOBT, sensitivity and specificity were 53.1% (17 neoplasms) and 59.4%, resulting in 8.1 colonoscopies/neoplasm. One I‐FOBT having ≥50 ngHb/mL of buffer provided equivalent sensitivity but 94.0% specificity, resulting in 2.1 colonoscopies/neoplasm. By analysing the higher of two I‐FOBTs at 50 ngHb/mL threshold, sensitivity increased to 68.8% (22 neoplasms, P = 0.063), specificity fell to 91.9% (P < 0.001), but still required 2.1 colonoscopies/neoplasm.

Conclusions  In this population, quantified I‐FOBT had significantly better specificity than G‐FOBT for significant neoplasms, reducing the number of colonoscopies needed/neoplasm detected. Results depend on the number of I‐FOBTs performed and the chosen development threshold.
Long‐term follow‐up of a population based cohort with monoclonal proteinaemia
Authors: Schaar Cees G. , Le Cessie Saskia , Snijder Simone , Franck Paul F. H. , Wijermans Pierre W. , Ong Cisca , Kluin‐Nelemans Hanneke
Published: November 2008

Prospective studies on the risk of malignant transformation in patients with monoclonal gammopathy of undetermined significance (MGUS) and factors predictive of survival are lacking. The Dutch Comprehensive Cancer Centre West, comprising 1·6 million inhabitants, initiated a prospective hospital‐based cohort study on 1464 patients with newly diagnosed M‐proteinaemia, median age 73 (17–103) years. M‐protein related diagnoses, patients’ characteristics, laboratory investigations, bone marrow examinations and skeletal X‐rays were registered with a yearly follow‐up. Main endpoints were death, or new diagnoses of multiple myeloma and non‐Hodgkin lymphoma. Kaplan–Meier survival curves were compared with age‐ and gender‐matched survival data from the total Dutch population. Cumulative malignant transformation was corrected for death using a competing risk model. Risk factors for transformation or death were analyzed by univariate and multivariate analyses. In 1007 MGUS‐patients, malignant transformation was associated with rising M‐protein levels, IgA and IgM isotype and occurred at a yearly rate of 0·4%. All MGUS patients survived less than a matched cohort of the Dutch population, even in the absence of M‐protein‐associated comorbidity. Serum albumin levels at entry appeared highly predictive for survival. M‐proteinaemia is not an innocent symptom. Although malignant transformation occurs rarely, survival is shortened irrespective of comorbidity.
Familial thrombocytosis caused by the novel germ‐line mutation p.Pro106Leu in the MPL gene
Authors: El‐Harith El‐Harith Abdelghaffar , Roesl Cornelia , Ballmaier Matthias , Germeshausen Manuela , Frye‐Boukhriss Hildegard , Von Neuhoff Nils , Becker Christian , Nürnberg Gudrun , Nürnberg Peter , Ahmed Mirghani Ali Mohamed , Hübener Jeannette , Schmidtke Jörg , Welte Karl , Stuhrmann Manfred
Published: November 2008

Familial thrombosis (FT) has been described as a rare autosomal‐dominant disorder, mostly caused by activating mutations of the thrombopoietin gene (THPO). Other cases of FT have been linked to one of two different germline mutations in the myeloproliferative leukaemia virus oncogene gene (MPL), which codes for the thrombopoietin receptor MPL. We studied an Arab family with two siblings with severe thrombocytosis by linkage analysis and obtained evidence for linkage to MPL. Sequencing revealed homozygosity for the novel MPL germline mutation p.Pro106Leu (c.317C > T) in the two siblings. Subsequently, homozygosity for p.Pro106Leu was identified in six further FT patients from three other Arab families. Of 18 heterozygous carriers, 14 had normal platelet counts, while four had mild thrombocytosis. Strong support for association of the novel MPL mutation p.Pro106Leu with development of familial thrombocytosis has been obtained. Overall, p.Pro106Leu was absent on 386 alleles of 193 healthy German controls and present on 14 of 426 alleles (3·3%) of 213 unrelated Arabs, which was statistically significantly different (P < 0·001, Fisher’s exact test). We assume that p.Pro106Leu is a frequent MPL mutation in the Arab population, leading to severe thrombocytosis in homozygotes and occasionally to mild thrombocytosis in heterozygotes. In the families described the mode of inheritance could be regarded as autosomal‐recessive with possible mild heterozygote manifestation rather than autosomal‐dominant with high penetrance as usually seen in FT.
Growth differentiation factor 15 production is necessary for normal erythroid differentiation and is increased in refractory anaemia with ring‐sideroblasts
Authors: Ramirez Jean‐Marie , Schaad Olivier , Durual Stephane , Cossali Dominique , Docquier Mylène , Beris Photis , Descombes Patrick , Matthes Thomas
Published: November 2008

The disturbed erythropoiesis in patients with refractory anaemia with ring‐sideroblasts (RARS) is characterized by intramedullary apoptosis of erythroid precursors and increased iron accumulation in mitochondria. To gain insight into these pathophysiological mechanisms we compared the gene expression profile (GEP) of erythroid precursors from RARS patients to the GEP of normal erythroid precursors. Three hundred sixty four probe sets were up‐, and 253 probe sets downregulated in RARS cells. Interestingly, Growth Differentiation factor 15 (GDF15), a cytokine from the TGFβ family, was dramatically upregulated in all RARS patients. Measurement of GDF15 in the sera from twenty RARS patients confirmed this finding by showing significantly, 7·2‐fold, increased protein levels (3254 ± 1400 ng/ml vs. 451 ± 87 ng/ml in normals). In vitro studies demonstrated erythroid‐specific production of GDF15 and dependence on erythropoietin. Induction of apoptosis by arsenic trioxide, a drug which acts via reduction of the mitochondrial membrane potential, also stimulated GDF15 production. Downregulation of endogenous GDF15 production in erythoblasts by specific siRNA led to diminished erythroid differentiation. Taken together, our findings demonstrate a new role for GDF15 in normal erythropoiesis as well as in the ineffective erythropoiesis of RARS patients.
Predicting response to immunosuppressive therapy and survival in severe aplastic anaemia
Authors: Scheinberg Phillip , Wu Colin O. , Nunez Olga , Young Neal S.
Published: November 2008

Horse anti‐thymocyte globulin (h‐ATG) and ciclosporin are the initial therapy for most patients with severe aplastic anaemia (SAA), but there is no practical and reliable method to predict response to this treatment. To determine whether pretreatment blood counts discriminate patients with SAA who have a higher likelihood of haematological response at 6 months to immunosuppressive therapy (IST), we conducted a single institution retrospective analysis on 316 SAA patients treated with h‐ATG‐based IST from 1989 to 2005. In multivariate analysis, younger age, higher baseline absolute reticulocyte count (ARC), and absolute lymphocyte count (ALC) were highly predictive of response at 6 months. Patients with baseline ARC ≥ 25 × 109/l and ALC ≥ 1 × 109/l had a much greater probability of response at 6 months following IST compared to those with lower ARC and ALC (83% vs. 41%, respectively; P < 0·001). This higher likelihood of response translated to greater rate of 5‐year survival in patients in the high ARC/ALC group (92%) compared to those with a low ARC/ALC (53%). In the era of IST, the baseline ARC and ALC together serve as a simple predictor of response following IST, which should guide in risk stratification among patients with SAA.
The relationship between lifestyle factors and venous thromboembolism among women: a report from the MISS study
Authors: Lindqvist Pelle G. , Epstein Elisabeth , Olsson Håkan
Published: November 2008

There has been a great advance in our knowledge of the role that thrombophilic factors play in the risk of venous thromboembolic events (VTE). However, the effect of lifestyle factors on VTE has been inadequately explored in large scale studies of women. This cohort study comprised one thousand native Swedish women for each age year between 25 and 64 inclusive (total = 40 000) drawn from the South Swedish population registry for 1990 (n = 40 000), who were followed for a mean of eleven years. Seventy‐four percent completed a questionnaire at the inception of the study (n = 29 518) and 24 098 women responded to a follow‐up inquiry between the years 2000–2002. The main outcome was the relationship between VTE and physical exercise, smoking habits, and alcohol consumption. Moderate drinkers of alcohol (10–15 g/d) and women engaged in strenuous exercise were at half the risk of VTE compared to those who consumed little or no alcohol or lived a sedentary life. Heavy smoking was associated with a 30% increased risk of VTE. Lifestyle factors have a major impact on the risk of VTE. Women non‐smokers who were physically active and who consumed alcohol in moderation were at a lower risk of VTE.
Clinical and laboratory phenotypes associated with the aspirin‐like defect: a study in 17 unrelated families
Authors: Rolf Nina , Knoefler Ralf , Bugert Peter , Gehrisch Siegmund , Siegert Gabriele , Kuhlisch Eberhard , Suttorp Meinolf
Published: November 2008

Aspirin‐like defect (ALD) is a rare, mostly autosomal dominant inherited dysfunction of the intraplatelet arachidonic acid (AA) pathway leading to impaired thromboxane A2 signalling. We aimed to establish diagnostic criteria for ALD diagnosis and present clinical and laboratory phenotypes of 52 individuals from 17 unrelated families. Platelet in vitro function was determined on the basis of platelet aggregation response (PAR) to AA, adenosine diphosphate, collagen and ristocetin as well as PFA‐100® closure times (CT). Using impaired PAR to AA (≤10%) as the mandatory diagnostic criterion, ALD could be confirmed in 17 patients. Subsequently, family members were investigated and among 35 individuals an additional 13 ALD patients as well as 4 individuals with mild ALD (PAR to AA: 19–32%) were identified. At least one bleeding symptom was reported by 25 (74%) ALD patients and prolonged CT was detected in 24 (71%) of the cases, both significantly correlated with impaired PAR to AA (P = 0·001 and P = 0·002, respectively). An estimated 0·6% prevalence was determined for ALD in our paediatric patients with suspected coagulation disorders. Due to the mild bleeding symptoms, ALD is probably underdiagnosed. If ALD is suspected, PAR to AA is suitable for the identification of individuals at risk of increased haemorrhage.
Expression and function of toll like receptors in chronic lymphocytic leukaemia cells
Authors: Muzio Marta , Scielzo Cristina , Bertilaccio Maria T. S. , Frenquelli Michela , Ghia Paolo , Caligaris‐Cappio Federico
Published: November 2008

Mature B‐cells can recognize microbial antigens via B‐cell‐receptor (BCR) in a specific way and via Toll‐like receptors (TLR) in a costimulatory manner. A wealth of information is gathering on the possible role of antigenic stimulation in the natural history of Chronic Lymphocytic Leukaemia (CLL). However little is known regarding the repertoire and function of TLR in CLL cells. The TLR family includes 10 different transmembrane proteins devoted to recognize specific pathogen‐associated molecular patterns and to alarm immunocompetent cells to trigger an immune response. Here, we studied fresh leukaemic cells for the expression pattern of TLR1 to TLR10, NOD1, NOD2 and SIGIRR (also known as TIR8). CLL cells were found to express several pattern recognition receptors including TLR1, TLR2, TLR6, TLR10, NOD1 and NOD2. The specific TLR expressed by CLL cells were functional. Leukaemic cells, upon stimulation with TLR1/2/6 ligands, such as bacterial lipopeptides, activated the nuclear factor‐κB signalling pathway, expressed CD86 and CD25 activation molecules, and were protected from spontaneous apoptosis. These findings further support the hypothesis that CLL cells resemble antigen‐activated B‐cells and suggest a potential role of TLR in modulating CLL cell response in the context of specific antigen recognition.
Haemoglobin F modulation in childhood sickle cell disease
Authors: Trompeter Sara , Roberts Irene
Published: November 2008

While supportive care remains the best option for most well children with sickle cell disease (SCD), increasing awareness of early signs of chronic organ damage in childhood has focused attention on therapy which modulates the natural history of the disease. Since cure by stem cell transplantation is only feasible for a minority and gene therapy remains developmental, pharmacological modification by Haemoglobin F (HbF)‐inducers, is the most widely used approach in SCD. Currently, the only HbF modulator with a clear place in the management of childhood SCD is hydroxycarbamide for which the main indications are frequent painful crises and recurrent acute chest syndrome. In the majority of SCD children treated with hydroxycarbamide there is clear evidence of clinical benefit and the drug is well tolerated. The main disadvantages are the need for frequent monitoring and uncertainity about long‐term risks of carcinogenicity and impaired fertility, although these risks appear to be very low. The role of hydroxycarbamide in sickle‐associated central nervous system disease remains to be established. Decitabine and butyrate derivatives show some promise although robust data in children with SCD are lacking. A number of other drugs are currently under investigation for their effects on HbF production including thalidomide and lenolidamide.
Amplification and invasiveness of epithelial progenitors during gastric carcinogenesis in trefoil factor 1 knockout mice
Authors: Karam S. M. , Tomasetto C. , Rio M.‐C.
Published: November 2008

Abstract.  Objective: It is not known whether or not epithelial progenitors of the pyloric antrum are involved in gastric carcinogenesis. Normally, these progenitors give rise to two main cell lineages: pit and gland mucous cells. This study was designed to examine the changes that occur in pyloric antral mucous cell lineages and their progenitors during development of gastric adenoma and carcinoma in trefoil factor 1 (TFF1) knockout mice. Materials and methods: Pyloric antral mucosal tissues of TFF1 knockout mice at ages from 3 days to 17 months were processed for histochemical analysis using Ulex europaeus and Grifforia simplifolica lectins as markers for pit and gland mucous cells, respectively. The dividing epithelial progenitors were identified by using immunohistochemical and electron microscopy techniques. Results: TFF1 loss was associated with amplification of both mucus‐secreting pit and gland cells. Both lectins examined bound not only to mature mucous cells, but also to most of epithelial progenitors which gradually amplified with age and frequently were seen in mitosis. Analysis of 12‐ to 17‐month‐old TFF1‐deficient stomachs revealed occasional groups of poorly differentiated mucosal cells with features similar to those of epithelial progenitors (or stem cells), in the basal portion of the antral mucosa. These cells eventually invaded the muscularis mucosa while maintaining some capacity to differentiate. Conclusion: This study shows that the progenitors of pit and gland mucous cells contribute to gastric carcinogenesis in the pyloric antrum of TFF1 knockout mice, strongly supporting the concept of stem cell origin of cancer.
Anti‐allergic effects of PG102, a water‐soluble extract prepared from Actinidia arguta, in a murine ovalbumin‐induced asthma model
Authors: Kim D. , Kim S. H. , Park E‐J. , Kang C‐Y. , Cho S‐H. , Kim S.
Published: November 2008

Background
Asthma is a chronic inflammatory disease of the lung and its incidence has been increasing around the world. We previously reported that oral administration of a water‐soluble extract prepared from Actinidia arguta, code‐named PG102, could modulate the level of Th1 and Th2 cytokines and suppress the production of immunoglobulin E (IgE) in the ovalbumin (OVA)‐immunized murine model as well as in the in vitro cell culture system, and furthermore could significantly improve dermatitis conditions in the NC/Nga murine model. These data suggested that PG102 might have therapeutic effects in a broad range of allergic diseases.

Objective
To assess the possible anti‐allergic effects of PG102 in the OVA‐induced murine asthma model.

Methods
The quality of PG102 was standardized, using its effects on the production of IgE, IL‐5, and IL‐13, in in vitro cell culture systems. To test effects on asthma, BALB/c mice were orally administrated with PG102, followed by OVA sensitization and challenge to induce asthmatic symptoms. Airway hyperresponsiveness (AHR), bronchoalveolar lavage fluid, serum, and lung tissue were analysed by using various methods.

Results
PG102 could decrease the level of IgE, IL‐5, and IL‐13 in in vitro cell culture systems with IC50 being 1.12–1.43 mg/mL. PG102 could ameliorate asthmatic symptoms, including AHR and eosinophilia in the lungs. Such improvement of asthmatic symptoms by PG102 was accompanied by the down‐regulation of IL‐5 and IgE. In PG102‐treated mice, high level expression of heme oxygenase‐1, a potent anti‐inflammatory enzyme, was observed in alveolar inflammatory cells, while the mRNA levels of foxp3, TGF‐β1, and IL‐10, important markers for regulatory T cells, were also up‐regulated in the lung tissue.

Conclusions
PG102 may have potential as a safe and effective reagent for the prevention or treatment of asthma.
Antibiotic use in the first year of life and risk of atopic disease in early childhood
Authors: Kusel M. M. H. , De Klerk N. , Holt P. G. , Sly P. D.
Published: November 2008

Background
Reduced post‐natal microbial stimulation resulting from improvements in public health measures, smaller family size, and through increased antibiotic use has been postulated to account for the increasing prevalence of atopic diseases seen predominantly in developed countries.

Objective
To investigate use of antibiotics in the first year of life and subsequent development of atopic disease in early childhood.

Methods
A prospective birth cohort of 198 children at high atopic risk was recruited prenatally and followed for 5 years. Illnesses and antibiotic use were ascertained through daily diaries, and diagnoses of asthma and hayfever were collected by questionnaire interviews. The children were examined regularly for eczema, and atopic status was defined by skin prick tests and serum total IgE. The effect of antibiotic use on subsequent atopic disease was examined using logistic regression with propensity score adjustment.

Results
54.0% (107/198) of children received at least one course of antibiotics, mainly for acute respiratory illnesses (ARI). Thirty‐three percent (329/984) of the ARI involved the lower respiratory tract (LRI). Twenty‐three percent (222/984) of ARI were treated with antibiotics, with LRI significantly more likely to receive antibiotics. Antibiotic use was associated with asthma (unadjusted odds ratio 2.3; 95% confidence interval 1.2–4.5; P=0.01) but this association was reduced after propensity score adjustment. No associations were found between antibiotic use and eczema, current wheeze, current asthma, atopic asthma, allergic rhinoconjunctivitis or atopy.

Conclusion
Although this was a small study, systematic and careful monitoring of ARI, antibiotic use, and asthma and atopic diseases did not indicate that receipt of antibiotics early in life led to subsequent asthma or atopy at 5 years.
5,7-dihydroxy-3,4,6-trimethoxyflavone inhibits the inflammatory effects induced byBacteroides fragilisenterotoxin via dissociating the complex of heat shock protein 90 and IκBα and IκB kinase-γ in intestinal epithelial cell culture
Authors: Kim J M , Lee D H , Kim J S , Lee J Y , Park H-G , Kim Y-J , Oh Y-K , Jung H C , Kim S I
Published: November 2008

Enterotoxin produced by enterotoxigenic Bacteroides fragilis (BFT) has been associated with mucosal inflammation and diarrhoeal diseases. In this study, the anti-inflammatory molecular mechanism of 5,7-dihydroxy-3,4,6-trimethoxyflavone (eupatilin) was characterized in an HT-29 intestinal epithelial cell line stimulated with BFT. Pre-treatment of HT-29 cells with eupatilin decreased the production significantly of both interleukin (IL)-8 and prostaglandin E2 induced by BFT in a dose-dependent manner. BFT-activated nuclear factor-kappaB (NF-κB) signals in HT-29 cells and pretreatment with eupatilin suppressed NF-κB activation that resulted in the significant inhibition of IL-8 and cyclo-oxygenase-2 expression. BFT-induced phosphorylation of both IκBα and IκB kinase (IKK) signals was prevented in eupatilin-pretreated HT-29 cells. Transfection of siRNA for IKK-α and IKK-β decreased the production of IL-8 and prostaglandin E2; however, the transfection of IKK-β siRNA showed a more significant reduction of BFT-induced IκBα phosphorylation compared with that of IKK-α siRNA. In addition, herbimycin A, a specific inhibitor of heat shock protein 90 (Hsp90), decreased the BFT-induced activation of IKK and NF-κB, suggesting that Hsp90 is associated with a pathway of IKK-NF-κB-IL-8/cyclo-oxygenase-2 gene signalling. Furthermore, eupatilin dissociated the complex between Hsp90 and IKK-γ in BFT-stimulated HT-29 cells. These results suggest that eupatilin can suppress the NF-κB signalling pathway by targeting the Hsp90-IKK-γ complex in intestinal epithelial cells and may attenuate BFT-induced inflammatory responses.
Mutations in the ABCC8 (SUR1 subunit of the KATP channel) gene are associated with a variable clinical phenotype
Authors: Klupa Tomasz , Kowalska Irina , Wyka Krystyna , Skupien Jan , Patch Ann‐Marie , Flanagan Sarah E. , Noczynska Anna , Arciszewska Malgorzata , Ellard Sian , Hattersley Andrew T. , Sieradzki Jacek , Mlynarski Wojciech , Malecki Maciej T.
Published: November 2008

Objective  Mutations in the ABCC8 gene encoding the SUR1 subunits of the β‐cell K‐ATP channel cause neonatal diabetes (ND) mellitus. We aimed to determine the contribution of ABCC8 gene to ND in Poland, to describe the clinical phenotype associated with its mutations and to examine potential modifying factors.

Patients  The Nationwide Registry of ND in Poland includes patients diagnosed before 6 months of age. In total 16 Kir6·2 negative patients with ND, 14 permanent and 2 relapsed transient, were examined.

Measurements  ABCC8 gene mutations were detected by direct sequencing. Mutation carriers’ characteristics included clinical data and biochemical parameters. In addition, we performed the hyperinsulinaemic euglycaemic clamp and tested for islet‐specific antibodies in diabetic subjects.

Results  We identified two probands with permanent ND (one heterozygous F132V mutation carrier and one compound heterozygote with N23H and R826W mutations) and two others with relapsed transient ND (heterozygotes for R826W and V86A substitutions, respectively). One subject, a heterozygous relative with the R826W mutation, had adult onset diabetes. There were striking differences in the clinical picture of the mutation carriers as the carrier of two mutations, N23H and R826W, was controlled on diet alone with HbA1c of 7·3%, whereas the F132V mutation carrier was on 0·66 IU/kg/day of insulin with HbA1c of 11·7%. The C‐peptide level varied from 0·1 ng/ml (F132V) to 0·75 ng/ml (V86A). We also observed a variable insulin resistance, from moderate (M = 5·5 and 5·6 mg/kg/min, respectively, in the two R826W mutation carriers) to severe (M = 2·6 mg/kg/min in the F132V mutation carrier). We were able to transfer two patients off insulin to sulphonylurea (SU) and to reduce insulin dose in one other patient. Interestingly, there was no response to SU in the most insulin resistant F132V mutation carrier despite high dose of glibenclamide. All examined auto‐antibodies were present in one of the subjects, the V86A mutation carrier, although this did not seem to influence the clinical picture, as we were able to transfer this girl off insulin.

Conclusion  Mutations in SUR1 are the cause of about 15% of Kir6·2 negative permanent ND in Poland. The clinical phenotype of SUR1 diabetic mutation carriers is heterogeneous and it appears to be modified by variable sensitivity to insulin.
Components of the renin‐angiotensin‐aldosterone system in plasma and ascites in hepatic cirrhosis
Authors: Kuiper J. J. , Boomsma F. , Van Buren H. , De Man R. , Danser A. H. J. , Van Den Meiracker A. H.
Published: November 2008

Background  Decompensated liver cirrhosis is characterized by activation of the renin‐angiotensin‐aldosterone system (RAAS). We investigated whether compartmentalization of these components occurs in ascitic fluid.

Methods  In 26 patients with cirrhosis RAAS components and albumin were quantified in simultaneously obtained plasma and ascitic fluid samples. Renin degradation was determined in vitro in plasma and ascites.

Results  Plasma angiotensinogen was below normal reference values in all but two patients and correlated inversely with plasma renin (r = –0·73, P < 0·001). Plasma renin activity was elevated in most subjects. The plasma and ascites concentrations of renin, prorenin, angiotensinogen and aldosterone were closely (P < 0·001) correlated. Expressed as a percentage of plasma levels, the angiotensinogen level (18 ± 11%) was slightly lower than the albumin level (23 ± 8%), whereas the aldosterone level (43 ± 18%) was considerably higher (P < 0·0001). For renin and prorenin these percentages were much lower (P < 0·0001), despite the fact that their molecular weight is lower than that of albumin and angiotensinogen. This was not due to a more rapid degradation of renin in ascites fluid, since the in‐vitro degradation rates of renin in plasma and ascitic fluid were identical.

Conclusion  In hepatic cirrhosis ascites can be regarded as an ultrafiltrate of plasma RAAS components. Since differences in molecular weight or metabolic rate cannot explain the low ascites‐to‐plasma ratio of renin and prorenin, either their transcapillary transport is impaired and/or they selectively bind to (pro)renin binding sites.
Parental history of myocardial infarction and risk of heart failure in male physicians
Authors: Djoussé L. , Gaziano J. M.
Published: November 2008

Background  Although heart failure (HF) remains a major public health issue, limited data are available on the utility of parental information on the risk of HF in offspring.

Materials and methods  We prospectively examined the association between parental history of myocardial infarction (MI) and incident HF among 20 187 offspring in the Physicians’ Health Study I. Parental history and age at MI was assessed by a questionnaire and a Cox regression was used to estimate relative risks of HF.

Results  After an average follow‐up of 19·6 years, 1036 new HF cases were documented. Overall, while a history of early parental MI (before age 55) was associated with a 32% increased risk of HF in offspring compared with subjects whose parent did not have MI, parental MI at older ages was not associated with HF risk. However, the relation between parental history of MI and HF was stronger and mainly observed for HF with antecedent MI. Compared with subjects without parental history of MI, multivariable adjusted hazard ratios (95% CI) for HF with antecedent MI were 3·44 (2·15–5·51), 2·24 (1·20–4·21), 1·26 (0·63–2·51), and 1·37 (0·92–2·03) for parental MI occurred at the age of < 55, 55–59, 60–64, and 65 + y, respectively.

Conclusions  Our data suggest that parental MI at an early age is a strong and independent predictor of HF with antecedent MI among US male physicians. This information, along with existing tools, may help clinicians identify patients at risk of HF with antecedent MI.
Biocompatibility of haemodialysis membranes determined by gene expression of human leucocytes: a crossover study
Authors: Wilflingseder J. , Perco P. , Kainz A. , Korbély R. , Mayer B. , Oberbauer R.
Published: November 2008

Background  Biocompatibility of haemodialysis membranes is the most important quality criteria to enable long‐term dialysis without major harmful effects. This study sought to evaluate the differences of genomic signatures derived from peripheral blood mononuclear cells (PBMC) in patients undergoing haemodialysis treatment using two different dialyser membranes: one semi‐synthetic and one full‐synthetic membrane.

Design  Microarray experiments were conducted in PBMCs of four stable haemodialysis patients before and after dialysis comparing semi‐synthetic (Hemophan® GFS Plus 16) and full‐synthetic (Hemoflow® FX80) dialysis membranes, respectively. Genes differentially expressed when comparing the two different membranes used were analysed in order to elucidate the underlying molecular mechanisms affecting PBMCs in the course of dialysis treatment.

Results  One hundred and seventy‐two genes were identified as up‐regulated after treatment with semi‐synthetic membranes when compared to full‐synthetic membranes. These genes could be assigned to processes including immunity and defence, signal transduction, and apoptosis. Dialysis with a full‐synthetic membrane, on the other hand, led to an activation of 72 genes that were mainly involved in cell cycle and cell cycle control.

Conclusion  The over‐representation of genes belonging to immunity/defence, signal transduction, and apoptosis as found with semi‐synthetic membranes suggests that full‐synthetic membranes are more biocompatible than semi‐synthetic membranes.
Prostaglandin receptors mediate effects of substances released from ischaemic rat hearts on non‐ischaemic cardiomyocytes
Authors: Birkenmeier K. , Janke I. , Schunck W.‐H. , Trimpert C. , Krieg T. , Landsberger M. , Völker U. , Felix S. B. , Staudt A.
Published: November 2008

Background After ischaemia and during reperfusion, rat hearts release cardiodepressive substances that are putatively cyclooxygenase‐2‐dependent. The present study analyses the mechanisms by which these substances mediate their effect downstream of cyclooxygenase‐2.

Materials and methods After 10 min of global stop‐flow ischaemia, isolated rat hearts were reperfused and post‐ischaemic coronary effluent was collected over a period of 30 s. Non‐ischaemic effluent collected before ischaemia was used as a control. We investigated the effect of the effluents on cell shortening and Ca++‐metabolism, by application of fluorescence microscopy of field‐stimulated adult rat cardiomyocytes incubated with fura‐2. Cells were pre‐incubated with inhibitors of protein kinase A and C and with antagonists of protein kinase A‐dependent prostaglandin receptors. We examined the expression of prostaglandin receptors in cardiomyocytes by Western blotting.

Results In contrast to non‐ischaemic effluent, post‐ischaemic effluent induced reduction of Ca++transient and cell shortening in the cardiomyocytes. In contrast to protein kinase C inhibitor Myr‐PKC [19–27], the protein kinase A inhibitor Rp‐cAMPS completely blocked the effect of post‐ischaemic effluent. Furthermore, we determined a cyclic adenosine monophosphate increase in cardiomyocytes that were pre‐incubated with post‐ischaemic effluent. The antagonist of prostaglandin E‐receptor EP2 AH6809 and the antagonist of receptor subtype EP4 AH23848 attenuated the effect of post‐ischaemic effluent in contrast to other antagonists of prostaglandin D and I receptors, which did not influence the effect. In lysates of adherend cardiomyocytes, expression of prostaglandin D, E and I receptors was detected by Western blotting.

Conclusions The effect of post‐ischaemic effluent is mediated by the protein kinase A‐dependent prostaglandin‐receptor subtypes EP2 and EP4 downstream of cyclooxygenase‐2.
Modelling the effects of ageing on Cd, Zn, Ni and Cu solubility in soils using an assemblage model
Authors: Buekers J. , Degryse F. , Maes A. , Smolders E.
Published: November 2008

Ageing reactions can reduce trace metal solubility and can explain natural attenuation of contaminated soils. We modelled ageing reactions in soil with an assemblage model that considers slow reactions in Fe‐oxyhydroxides and reversible sorption on organic matter and clay minerals. Metal adsorption kinetics on Fe‐oxyhydroxides was obtained from data with synthetic oxyhydroxides. Metal solubility and isotopic exchangeability data were obtained from 28 soils amended with Ni, Zn, Cu and Cd metal salts and monitored for 850 days. The assemblage model was constructed in WHAM 6.0 and used soil properties and dissolved organic matter as input data. The model was first validated to predict dissolved metal concentrations, based on the concentration of isotopic exchangeable metals. The model overestimated metal solubility without parameter adjustment by mean factors of 4–7, and successful fits were obtained by increasing the specific surface area of Fe‐oxyhydroxides from measured values of synthetic systems to a value of 600 m2 g−1 recommended by other authors. The effect of ageing on the isotopic exchangeable metal fraction was subsequently modelled starting from the predicted fraction of metals present on Fe‐oxyhydroxides immediately after soil spiking. The observed isotopic exchangeable metal fractions of Ni, Zn and Cd agreed reasonably well with predicted values. The model predicts that ageing reactions are more pronounced at higher pH because metal sorption is increasingly directed to oxyhydroxide surfaces with increasing soil pH. Modelling fixation of Cu requires more information on fixation of that metal in organic matter.