bluuebunny/crossref_metadata_2025_split huggingface dataset:
Polyarginine as a multifunctional fusion tag
Authors: Fuchs Stephen M. , Raines Ronald T. 
Published: June 2005

AbstractFusion to cationic peptides, such as nonaarginine (R9), provides a means to deliver molecular cargo into mammalian cells. Here, we provide a thorough analysis of the effect of an R9tag on the attributes of a model protein: bovine pancreatic ribonuclease (RNase A). The R9tag diminishes the conformational stability of RNase A (ΔTm=−8°C in phosphate‐buffered saline). This effect is nearly mitigated by the addition of salt. The tag does not compromise the enzymatic activity of RNase A. An R9tag facilitates the purification of RNase A by cation‐exchange chromatography and enables the adsorption of RNase A on glass slides and silica resin with the retention of enzymatic activity. The tag can be removed precisely and completely by treatment with carboxypeptidase B. Finally, the R9tag increases both the cellular uptake of RNase A and the cytotoxicity of G88R RNase A, a variant that evades the cytosolic ribonuclease inhibitor protein. Thus, we conclude that polyarginine is a versatile protein fusion tag.
Crystal structure of the protease‐resistant core domain of Yersinia pestis virulence factor YopR
Authors: Schubot Florian D. , Cherry Scott , Austin Brian P. , Tropea Joseph E. , Waugh David S. 
Published: June 2005

AbstractYersinia pestis, the causative agent of the plague, employs a type III secretion system (T3SS) to secrete and translocate virulence factors into to the cytoplasm of mammalian host cells. One of the secreted virulence factors is YopR. Little is known about the function of YopR other than that it is secreted into the extracellular milieu during the early stages of infection and that it contributes to virulence. Hoping to gain some insight into the function of YopR, we determined the crystal structure of its protease‐resistant core domain, which consists of residues 38–149 out of 165 amino acids. The core domain is composed of five α‐helices that display unexpected structural similarity with one domain of YopN, a central regulator of type III secretion in Y. pestis. This finding raises the possibility that YopR may play a role in the regulation of type III secretion.
Palsy of the rear limbs in Mycobacterium lepraemurium-infected mice results from bone damage and not from nerve involvement
Authors: Rojas-Espinosa O , Becerril-Villanueva E , Wek-Rodríguez K , Arce-Paredes P , Reyes-Maldonado E 
Published: June 2005

Abstract
A small but relatively constant proportion (3–5%) of mice chronically infected with Mycobacterium lepraemurium (MLM) develops bilateral paralysis of the rear limbs. The aim of the study was to investigate whether or not the bilateral leg palsy results from nerve involvement. Direct bacterial nerve infection or acute/delayed inflammation might possibly affect the nerves. Therefore, palsied animals were investigated for the presence of: (a) histopathological changes in the leg tissues including nerves, bones and annexes, and (b) serum antibodies to M. lepraemurium and M. leprae lipids, including phenolic glycolipid I from M. leprae. Histopathological study of the palsied legs revealed that the paralysis was not the result of direct involvement of the limb nerves, as neither bacilli nor inflammatory cells were observed in the nerve branches studied. Antibodies to brain lipids and cardiolipin were not detected in the serum of the palsied animals, thus ruling out an immune response to self-lipids as the basis for the paralysis. Although high levels of antibodies to MLM lipids were detected in the serum of palsied animals they were not related to limb paralysis, as the nerves of the palsied legs showed no evidence of inflammatory damage. In fact, nerves showed no evidence of damage. Paralysis resulted from severe damage of the leg bones. Within the bones the bone marrow became replaced by extended bacilli-laden granulomas that frequently eroded the bone wall, altering the normal architecture of the bone and its annexes, namely muscle, tendons and connective tissue. Although this study rules out definitively the infectious or inflammatory damage of nerves in murine leprosy, it opens a new avenue of research into the factors that participate in the involvement or the sparing of nerves in human and murine leprosy, respectively.
Leaky phenotype of X-linked agammaglobulinaemia in a Japanese family
Authors: Kaneko H , Kawamoto N , Asano T , Mabuchi Y , Horikoshi H , Teramoto T , Matsui E , Kondo M , Fukao T , Kasahara K , Kondo N 
Published: June 2005

SummaryX-linked agammaglobulinaemia (XLA) is an inherited immunodeficiency that is caused by a block in early B-cell differentiation. Whereas early B precursors in the bone marrow are present in substantial numbers, XLA-affected individuals have dramatically reduced numbers of circulating mature B cells, plasma cells and immunoglobulins of all isotypes. We report on a Japanese family with 3 XLA patients, in whom the serum immunoglobulin levels and number of B cells showed a significant difference among them in spite of harbouring the same splice donor site mutation in the BTK gene. We developed concise method for detection of this mutation, which is helpful for discovering the carrier. Patient 2 showed a significant serum immunoglobulin levels of all isotypes, including allergen-specific IgE. Expression of a normal and truncated size BTK gene was detected in patient 2′s peripheral blood mononuclear cells (PBMCs). Expression of BTK protein was also detected in some B cells. These results suggest that the leaky phenotype in patient 2 was caused in part by the expression of a normal BTK gene transcript. The increased frequency of infection with age expanded the number of B cells with normal BTK gene expression and produced the serum immunoglobulin, including IgE.
NewLactobacillus acidophilusisolates reduce the release of leptin by murine adipocytes leading to lower interferon-γ production
Authors: Bleau C , Lamontagne L , Savard R 
Published: June 2005

SummaryLeptin produced by adipocytes increases Th1-dependent immunostimulation and autoimmune diseases. Lactobacilli are known to promote or suppress Th1 responses according to the isolates. We have investigated whether the sensitivity of Suriss Jim Lambert (SJL) mice to Th1-dependent immune diseases, when compared with C57BL/6 mice, may be modulated by selected lactobacilli able to decrease leptin release by adipocytes. White adipocytes were isolated from both C57BL/6 and SJL mice and incubated with bacterial extracts from new CBA4P and TPA3P isolates of Lactobacillus acidophilus and L. rhamnosus 9595 (LR), or with conditioned media (CM) from lactobacillus-treated macrophages. Immunomodulation induced by supernatants of treated adipocytes was determined by metabolic activity of syngenic splenic lymphocytes. Leptin produced by adipocytes, tumour necrosis factor (TNF)-α and interleukin (IL)-1β by macrophages, and IFN-γ and IL-4 by lymphocytes were quantified by enzyme-linked immunosorbent assay (ELISA) tests. Results revealed that supernatants from CBA4P- and LR-treated adipocytes decreased the metabolic activity of lymphocytes from SJL mice, whereas adipocytes incubated with CM from CBA4P-treated macrophages showed no stimulation of lymphocytes. Such effects correlated with leptin levels. Lower levels of leptin were produced by adipocytes from SJL mice in the presence of CBA4P and LR extracts. Lymphocytes from SJL mice produced low levels of IFN-γ when incubated with supernatants from CBA4P-treated cells. Such immunosuppressive effects were dependent on levels of TNF-α and IL-1β produced by lactobacillus-treated macrophages. Taken together, these results suggest that the CBA4P isolate reduces levels of leptin in SJL mice, leading to lower IFN-γ production. Therefore, the CBA4P isolate of L. acidophilus is a promising new probiotic strain for the control of Th1 inflammatory diseases.
The interaction between staphylococcal superantigen-like proteins and human dendritic cells
Authors: Al-Shangiti A M , Nair S P , Chain B M 
Published: June 2005

Summary
Staphylococcus aureus produce a family of exotoxins (staphylococcal superantigen like proteins, SSLs) with structural, but not functional, homology to superantigens. These proteins have previously been shown to interact selectively with antigen presenting cells, including dendritic cells. The functional consequences of this interaction are now explored. SSL7 and 9 had no effect on viability or morphology of dendritic cells. The proteins did not induce dendritic cell maturation, as measured by cell surface phenotype. Exposure to SSL did not alter the ability of dendritic cells to take up FITC-dextran. Finally, exposure to SSLs did not impair the ability of the dendritic cells to stimulate allogeneic or antigen specific T cell responses. However, dendritic cells loaded with SSL7 or 9 were able to stimulate a T cell proliferative response in 3/8 healthy individuals tested. Sera from nine out of 10 individuals tested contained antibodies against both SSL7 and SSL9, and the response to each SSL was specific and not cross-reactive. The results demonstrate that SSLs are immunogenic in humans at both the B and T cell level, but it remains unclear whether this response is to the benefit of the bacterium or the host.
ProbioticLactobacillus-induced improvement in murine chronic inflammatory bowel disease is associated with the down-regulation of pro-inflammatory cytokines in lamina propria mononuclear cells
Authors: Matsumoto S , Hara T , Hori T , Mitsuyama K , Nagaoka M , Tomiyasu N , Suzuki A , Sata M 
Published: June 2005

SummaryIL-6/STAT-3 signals play key roles in inflammatory bowel disease (IBD). It is known that Lactobacillus casei strain Shirota (LcS) improves inflammatory disorders. This study aimed to elucidate the effect of LcS on murine chronic IBD and to clarify the mechanism. We focused the inhibitory effect of LcS on the production of IL-6 in lipopolysaccharide (LPS)-stimulated large intestinal lamina propria mononuclear cells (LI-LPMC) isolated from mice with chronic colitis and in RAW264.7 cells in vitro. We also determined in vivo the effect of LcS on murine chronic IBD models induced with dextran sodium sulphate and SAMP1/Yit mice. Finally, we examined the cellular determinants of LcS for the down-regulation of IL-6 secretion by LI-LPMC, RAW264.7 cells and peripheral blood mononuclear cells (PBMC) derived from patients with ulcerative colitis (UC). LcS, but not other strains of Lactobacillus, inhibited the production of IL-6 in LPS-stimulated LI-LPMC and RAW264.7 cells, down-regulating the nuclear translocation of NF-κB. The LcS-diet-improved murine chronic colitis is associated with the reduction of IL-6 synthesis by LI-LPMC. LcS also improved chronic ileitis in SAMP1/Yit mice. The release of IL-6 in vitro in LPS-stimulated LI-LPMC, RAW 264.7 cells and UC-PBMC was inhibited by a polysaccharide-peptidoglycan complex (PSPG) derived from LcS. This probiotic-induced improvement in murine chronic inflammatory bowel disease is associated with the down-regulation of pro-inflammatory cytokines such as IL-6 and IFN-γ production in LPMC. Therefore, LcS may be a useful probiotic for the treatment of human inflammatory bowel disease.
Pneumococcal conjugate vaccine followed by pneumococcal polysaccharide vaccine; immunogenicity in patients with ataxia-telangiectasia
Authors: Stray-Pedersen A , Aaberge I S , Früh A , Abrahamsen T G 
Published: June 2005

Summary
The immunodeficiency in Ataxia-telangiectasia (A-T) is characterised by low T and B cell counts, low levels of IgE, IgA and/or IgG2, and especially low levels of pneumococcal antibodies. The 23-valent pneumococcal polysaccharide vaccine (PPV23) has previously been shown not to be effective in A-T, but these patients are capable of making protective antibodies to other vaccines such as diphtheria and tetanus toxin, promising effect of the seven-valent pneumococcal conjugated vaccine (PCV7).
Nine A-T patients and 25 age and sex matched controls were vaccinated with both PCV7 and PPV23, and three A-T patients were vaccinated with PCV7 only. In the A-T patients, no significant increase in pneumococcal antibody levels were observed after the single PCV7, while the subsequent PPV23 vaccination resulted in a significant increase in antibody levels to the PPV23 mix, as well as to serotype 4, 14, 19F and to the geometric mean of serotype 4, 6B, 14, 18C, 19F, 23F which increased from median 0·2 (range 0·1–0·5) microg/mL to 0·6 (0·2–1·5) microg/mL (P= 0·014). Compared to the patients’ baseline levels, the vaccinations induced a 1·5- to 7-fold increase in antibodies to the six different serotypes tested. The increases in pneumococcal antibody titres were lower than those observed in the controls (9- to 34-fold increase). The results are valuable in planning the care of A-T patients, using PCV7 to trigger and PPV23 to booster the immune response and possibly prevent severe pneumococcal disease.
Interferon-γ and tumour necrosis factor-α production by CD4+ T and CD8+ T lymphocytes in AIDS patients with tuberculosis
Authors: De Castro Cunha R M , Kallas E G , Rodrigues D S , Nascimento Burattini M , Salomao R 
Published: June 2005

SummaryTuberculosis (TB) is usually more severe in HIV-infected patients, and the immune derangement found in co-infected patients may differ from that in each isolated disease. Following mitogen stimulation of peripheral blood mononuclear cells (PBMC), interferon (IFN)-γ and tumour necrosis factor (TNF)-α production was evaluated in T cells by flow cytometry, and in culture supernatants by enzyme-linked immunosorbent assay (ELISA) in 33 individuals: 11 AIDS patients with tuberculosis, six asymptomatic HIV-1-infected patients, eight patients with tuberculosis and eight healthy controls. The proportion of CD4+ T lymphocytes expressing IFN-γ did not differ between the groups, whereas a trend towards increased proportions of TNF-α-expression in CD4+ T cells was observed in the TB compared to the HIV group, while intermediate values were observed in co-infected patients. Detection of IFN-γ and TNF-α in CD8+ T lymphocytes was higher in TB than in HIV individuals. Co-infected patients presented intermediate values for IFN-γ, while TNF-α detection was similar to that in HIV mono-infection. In conclusion, the proportion of T cells expressing IFN-γ was relatively preserved in co-infected patients compared to TB patients, while the percentage of T cells expressing TNF-α was decreased, mainly in CD8+ T lymphocytes. However, the marked reduction in T lymphocyte numbers in co-infected patients led to a striking reduction of both cytokines in PBMC supernatants, a finding that is consistent with the impaired response to Mycobacterium tuberculosis.
Epitope analysis of insulin autoantibodies using recombinant Fab
Authors: Padoa C J , Crowther N J , Thomas J W , Hall T R , Bekris L M , Torn C , Landin-Olsson M , Ortqvist E , Palmer J P , Lernmark Å , Hampe C S 
Published: June 2005

SummaryAutoantibodies to insulin are often the first autoantibodies detected in young children with type 1 diabetes and can be present before the onset of clinical diabetes. These autoantibodies and their epitopes are, however, not well characterized. We explored the use of monoclonal antibodies and their recombinant Fab as reagents for epitope analysis. In this study we cloned and characterized the recombinant Fab of the insulin-specific monoclonal antibody CG7C7. We found the epitope of this antibody to be located predominantly at the A-chain loop of the insulin molecule. The recombinant Fab was then used to compete for insulin binding against insulin autoantibodies present in sera from patients with type 1 or type 1·5 diabetes. In competition experiments with sera positive for autoantibodies to insulin the recombinant Fab significantly reduced the binding to [125I]-insulin by sera of type 1 (n = 35) and type 1·5 diabetes [latent autoimmune diabetes in adults (LADA)] (n = 14) patients (P &lt; 0·0001). We conclude that competition between insulin-specific monoclonal antibodies or their recombinant Fab and insulin autoantibodies should prove useful in the epitope analysis of autoantibodies to insulin.
Distinct gene expression patterns of peripheral blood cells in hyper–IgE syndrome
Authors: Tanaka T , Takada H , Nomura A , Ohga S , Shibata R , Hara T 
Published: June 2005

SummaryHyper-immunoglobulin E (IgE) syndrome (HIES) is one of the primary immunodeficiency syndromes. Although the cytokine dysregulation is suggested to play a role in its pathophysiology, the causative gene has not yet been identified. To investigate the pathophysiology and candidate genes involved in this disease, we performed microarray analysis of unstimulated peripheral CD4+ T cells and CD14+ cells, as well as peripheral blood mononuclear cells (PBMNC) stimulated with Staphylococcus aureus isolated from HIES patients and healthy controls. By microarray analysis, 38 genes showed over 2-fold differences between the HIES patients and healthy controls in purified CD14+ cells, although only small differences in the gene expression profiles were observed between the two groups in purified CD4+ T cells. RGC32 expression levels showed the greatest difference between the two groups, and were significantly  elevated  in  HIES  compared  with  those  in  severe  atopic  dermatitis or healthy controls using real-time PCR. A significantly larger number of lysosome-related genes were up-regulated, and significantly larger number of genes related to cell growth and maintenance were down-regulated in HIES. After the stimulation of PBMNC with Staphylococcus aureus, 51 genes showed over 3-fold differences between HIES patients and healthy controls. A significantly large number of immunoglobulin-related genes were up-regulated in HIES. The distinct patterns of gene expression profiles and RGC32 expression levels will be useful for understanding the pathophysiology and for diagnosis of HIES, respectively.
Galectin‐3 is not useful in thyroid FNA
Authors: Mills L. J. , Poller D. N. , Yiangou C. 
Published: May 2005

Introduction:  Previous studies have suggested that galectin‐3 immunohistochemistry may be useful in the fine needle aspiration (FNA) diagnosis of thyroid carcinoma as it has been reported to selectively stain carcinomas and not adenomas or goitres.Methods:  Fifty‐one patients were included in a prospective study of galectin‐3 in thyroid FNA; 88.2% were female and 11.8% male, mean age 53 years, range 25–87 years. Cell blocks were prepared and stained for galectin‐3 if any cells were present in needle washings from the respective FNAs.Results:  Twelve of 51 (23.5%) of cell blocks contained epithelial cells. One benign and one inadequate FNA were negative for galectin‐3 staining. One of five non‐diagnostic FNA cases, a papillary carcinoma on final histology showed positive staining. Four follicular neoplasm/suspicious of carcinoma cases showed negative staining. One malignant FNA case, a papillary carcinoma showed positive staining with galectin‐3 but three further carcinomas, two papillary and one follicular were galectin‐3 negative.Conclusion:  Galectin‐3 immunohistochemistry does not appear to be a useful adjunct to diagnosis in thyroid FNA as it does not reliably distinguish malignant and benign lesions. Many thyroid aspirates are of low cellularity and are not suitable for cell block immunohistochemistry.
Use of scaled forms of the infiltration equation for the estimation of unsaturated soil hydraulic properties (the Beerkan method)
Authors: Braud I. , De Condappa D. , Soria J. M. , Haverkamp R. , Angulo‐Jaramillo R. , Galle S. , Vauclin M. 
Published: June 2005

SummaryWater movement in soil can be described accurately at the local scale, provided that soil hydraulic properties can be determined with precision. Traditional methods for characterizing soil are often time consuming, and large areas cannot be sampled easily. We present a simple method for overcoming these difficulties. It is easy to implement and cheap. It is known as the Beerkan method, and it relies on particle‐size analysis, dry bulk density and simple infiltration tests in cylinders. We describe the experimental protocol and the method of data analysis, leading to the estimation of parameters describing hydraulic properties. Shape parameters depend on soil texture and are derived from particle‐size data. Normalization parameters depend on soil structure. They are derived by inverse modelling and optimization from the infiltration tests. The theoretical background relies on the sorptivity concept and scaled forms of the infiltration equation. The formalism for one‐ and three‐dimensional analysis is described. We assess the accuracy of the method using published data and simulated values, showing the soundness of the approach. For the purpose of illustration, we implemented a simple optimization technique on two bounding cases.
Structural differences between bulk and rhizosphere soil
Authors: Whalley W. R. , Riseley B. , Leeds‐Harrison P. B. , Bird N. R. A. , Leech P. K. , Adderley W. P. 
Published: June 2005

SummaryThe physical characteristics of the soil at the root–soil interface are crucial because they determine both physical aspects of root function such as water and nutrient uptake and the microbial activity that is most relevant to root growth. Because of this we have studied how root activity modifies the structure and water retention characteristic of soil adjacent to the root for maize, wheat and barley. These plants were grown in pots for a 6‐week growth period, then the soil adjacent to the root (rhizosphere soil) and bulk soil aggregates were harvested. These soil aggregates were then saturated and equilibrated at matric potentials between −600 kPa and saturation, and the water retention characteristics were measured. From subsamples of these aggregates, thin sections were made and the porosity and pore‐size distributions were studied with image analysis. Both image analysis and estimates of aggregated density showed that the rhizosphere soil and bulk soil had similar porosities. Growing different plants had a small but significant effect on the porosity of the soil aggregates. Image analysis showed that for all the plant species the structure of the rhizosphere soil was different to that of the bulk soil. The rhizosphere soil contained more larger pores. For maize and barley, water retention characteristics indicated that the rhizosphere soil tended to be drier at a given matric potential than bulk soil. This effect was particularly marked at greater matric potentials. The difference between the water retention characteristics of the bulk and rhizosphere soil for wheat was small. We compare the water retention characteristics with the data on pore‐size distribution from image analysis. We suggest that differences in wetting angle and pore connectivity might partly explain the differences in water retention characteristic that we observed. The impact of differences between the water retention properties of the rhizosphere and bulk soil is discussed in terms of the likely impact on root growth.