bluuebunny/crossref_metadata_2025 huggingface dataset:
IL‐12p40‐independent induction of protective immunity upon multiple Plasmodium berghei irradiated sporozoite immunizations
Authors: ROMERO J. F. , IBRAHIM G. H. , RENGGLI J. , HIMMELRICH H. , GRABER P. , CORRADIN G.
Published: October 2007

Both IFN‐γ and IL‐12 play critical roles in defence against malaria. In a previous study, using Plasmodium yoelii model, C57BL/6 IFN‐γ receptor deficient mice (IFN‐γR−/–) failed to develop protective immunity after a single immunization with irradiated sporozoites, but were protected after multiple immunizations. In contrast, in another study, BALB/c IFN‐γ gene knockout mice (IFN‐γ−/–) and BALB/c IL‐12‐deficient mice (IL‐12p40−/– and IL‐12p35−/–) were unable to mount protective immune response even after multiple immunizations with the same irradiated parasites. To better define the role of IFN‐γ and IL‐12p40 in sterile protection, we selected the C57BL/6 model. Wild‐type and IL‐12p40−/– mice were immunized with a single or multiple doses of P. berghei irradiated sporozoites. While the wild‐type mice were able to rapidly produce IFN‐γ and mount a protective immune response after a single immunization with irradiated sporozoites, IL‐12p40−/– mice were neither able to produce IFN‐γ nor were protected. However, both strains of mice were able to produce IFN‐γ and were protected after three doses of irradiated sporozoites. Protection was partially and largely mediated by CD4+ T cells and CD8+ T cells, respectively. Thus, IL‐12p40 plays an important role in mediating early protection by irradiated sporozoite immunization but is dispensable for protective immunity induced by several immunizations with irradiated sporozoites. Moreover, treatment of hyperimmune IL‐12p40−/– mice with rhIL‐18 bp‐Fc, an inhibitor of IL‐18 activity, did not abrogate protection indicating that IL‐18 is not required for the effector phase of the immune response; it remains possible, however, that IL‐18 may compensate for the lack of IL‐12p40 in the induction phase of the immune response.
Comparison of immune responses in mice infected with different strains of Strongyloides venezuelensis
Authors: MACHADO E. R. , UETA M. T. , LOURENÇO E. V. , ANIBAL F. F. , ROQUE‐BARREIRA M. C. , FACCIOLI L. H.
Published: October 2007

In human hosts and in murine models, the immune response to Strongyloides spp. is Th2 type. In addition, the profile of the host immune response follows various symptoms induced by Strongyloides spp. In the present study, we demonstrated that the L2 and L49 strains of Strongyloides venezuelensis obtained from Bolomys lasiurus and Nectomys squamipes induced significant and similar increases in eosinophil/mononuclear cell counts in the blood, peritoneal cavity fluid and bronchoalveolar lavage fluid when compared with uninfected mice. However, in the first 3 days of infection, IL‐4, IL‐5 and IFN‐γ levels were higher in the lungs of mice infected with the L2 strain, which also presented greater production of IgG and IgG1 than did mice infected with the L49 strain. The higher antibody and cytokine levels induced by the L2 strain correlated with a decrease in the number of female parasites recovered in the faeces of mice on post‐infection day 7. The results demonstrate that the L2 strain was a more potent stimulant of the humoral immune response, which can result in more efficient antibody‐dependent cell‐mediated cytotoxicity, a mechanism involved in eosinophil activation and parasite elimination. Further studies are needed in order to elucidate the molecular differences among parasites.
Determinants of memory in experimental filarial infections in mice
Authors: DASH Y. , RAMESH M. , GREINER D. , SHULTZ L. D. , KLEI T. R. , RAJAN T. V.
Published: October 2007

In this communication, we examine the determinants and duration of memory responses against filarial parasites using an intraperitoneal mouse model of Brugia pahangi infection. We assessed the role of T cells in the memory response against B. pahangi larvae by transferring splenic T cells from wild‐type mice primed with L3 into T‐cell‐deficient mice. We found that mice reconstituted with primed T cells cleared intraperitoneal infections with infective larvae in an accelerated manner. To determine the components that may be responsible for the memory response, we transferred unfractionated T cells or purified CD4+ T cells or CD8+ T cells from BALB/cByJ mice primed a month earlier with L3 into T‐cell‐deficient BALB/c TCRβ−/– mice. Recipients were challenged 10 days after adoptive transfer. Our data demonstrated that while either CD4+ or CD8+ T cells are able to confer some level of protection, both are required for an optimal recall response. To evaluate the longevity of the memory response, we primed several groups of wild‐type mice at different times over a year. These mice were then challenged with a single injection of B. pahangi L3. The gap between the priming and second injections of larvae ranged between 4 and 60 weeks. We found that the memory responses in BALB/cByJ mice lasted over a year whereas those in C57BL/6 mice waned more rapidly.
The role of TNF‐α in Trichuris muris infection I: influence of TNF‐α receptor usage, gender and IL‐13
Authors: HAYES K. S. , BANCROFT A. J. , GRENCIS R. K.
Published: October 2007

Th1 and Th2 responses to the gut‐dwelling nematode Trichuris muris have been well established in mouse models of infection, with Th2 responses clearly playing an important role in resistance. TNF‐α has previously been shown to play an undefined role in resistance, although it is not a typical Th2 cytokine. However, the relative importance of the two TNF‐α receptors, p55 and p75, has not previously been investigated. We demonstrate that p55 is the dominant TNF‐α receptor during T. muris infection as p55−/– mice are more susceptible to infection than p75−/– mice. Moreover, p75 clearly plays a role in negatively regulating TNF‐α. We also demonstrate that a gender difference influences the immune response of p55−/– and p75−/– mice in response to T. muris infection, with female mice fully expelling by day 35 post‐infection (p.i.) and male mice harbouring chronic infections. Further, this gender difference can be reversed with recombinant IL‐13 (rIL‐13) in male gene‐deficient mice or IL‐13R2.Fc treatment in female gene‐deficient mice.
Chlororespiration is involved in the adaptation of Brassica plants to heat and high light intensity
Authors: DÍAZ MILAGROS , DE HARO VIRGINIA , MUÑOZ ROMUALDO , QUILES MARÍA JOSÉ
Published: October 2007

Two species of Brassica were used to study their acclimation to heat and high illumination during the first stages of development. One, Brassica fruticulosa, is a wild species from south‐east Spain and is adapted to both heat and high light intensity in its natural habitat, while the other, Brassica oleracea, is an agricultural species that is widely cultivated throughout the world. Growing Brassica plants under high irradiance and moderate heat was seen to affect the growth parameters and the functioning of the photosynthetic apparatus. The photosystem II (PSII) quantum yields and the capacity of photosynthetic electron transport, which were lower in B. fruticulosa than in B. oleracea, decreased in B. oleracea plants when grown under stress conditions, indicating inhibition of PSII. However, in B. fruticulosa, the values of these parameters were similar to the values of control plants. Photosystem I (PSI) activity was higher in B. fruticulosa than in B. oleracea, and in both species this activity increased in plants exposed to heat and high illumination. Immunoblot analysis of thylakoid membranes using specific antibodies raised against the NDH–K subunit of the thylakoidal NADH dehydrogenase complex (NADH DH) and against plastid terminal oxidase (PTOX) revealed a higher amount of both proteins in B. fruticulosa than in B. oleracea. In addition, PTOX activity in plastoquinone oxidation, and NADH DH activity in thylakoid membranes were higher in the wild species (B. fruticulosa) than in the agricultural species (B. oleracea). The results indicate that tolerance to high illumination and heat of the photosynthetic activity was higher in the wild species than in the agricultural species, suggesting that plant adaptation to these stresses in natural conditions favours subsequent acclimation, and that the chlororespiration process is involved in adaptation to heat and high illumination in Brassica.
Manipulation of light and CO2 environments of the primary leaves of bean (Phaseolus vulgaris L.) affects photosynthesis in both the primary and the first trifoliate leaves: involvement of systemic regulation
Authors: ARAYA TAKAO , NOGUCHI KO , TERASHIMA ICHIRO
Published: October 2007

Possible involvement of systemic regulation of the photosynthetic properties of young leaves by the local environments and/or photosynthate production of the mature leaves were examined using Phaseolus vulgaris plants. When primary leaves (PLs) were treated with air containing 150 µL CO2 L−1 with the other plant parts in ambient air at a photosynthetic photon flux density (PPFD) of 300 µmol photon m−2 s−1, decreases in the photosynthetic rate measured at 360 µL CO2 L−1 and a PPFD of 300 µmol photon m−2 s−1 (A360) were markedly retarded in both PLs and the first trifoliate leaves (TLs) as compared to plants treated with 400 µL CO2 L−1. Conversely, when PLs were treated with 1000 µL CO2 L−1, decreases in A360 were accelerated in both PLs and TLs. Shading of PLs accelerated the decrease in PL A360, and delayed the decrease in TLs. In the CO2 treatments, changes in A360 in TLs were mainly attributed to the changes in ribulose bisphosphate (RuBP) carboxylation rate, while the shading of PLs caused increases in both the RuBP carboxylation and regeneration rates in TLs. The ribulose 1·5‐bisphosphate carboxylase/oxygenase (Rubisco) activity on chlorophyll basis, an indicator of sun/shade acclimation, differed both among PLs and among TLs in accordance with the redox state of photosystem II (PSII) in PLs. Although carbohydrate contents of TLs were not affected by any manipulation of PLs, changes in the photosynthetic capacities of TLs acted to compensate for changes in PL photosynthesis. These results clearly indicate that the CO2 and shade treatments of PLs not only affect photosynthetic properties of the PLs themselves, but also systemically affected the photosynthetic properties of TLs. Possible roles of the redox state and photosynthate concentration in PLs in regulation of photosynthesis in PLs and TLs are discussed.
The relationship between rhizosphere nitrification and nitrogen‐use efficiency in rice plants
Authors: LI YI LIN , FAN XIAO RONG , SHEN QI RONG
Published: October 2007

Two different rice cultivars, Yangdao 6 [Indica rice cultivar with high nitrogen‐use efficiency (NUE)] and Nongken 57 (Japonica rice cultivar with low NUE) were used to study the relationship between NUE and nitrification activity in the rice seedling rhizosphere soil using a rhizobox with three compartments, and a soil‐slicing method. The roots of both rice cultivars developed aerenchyma tissue [expressed as percentage porosity of root (POR)], but Yangdao 6 showed better development than Nongken 57. This root morphology change results in more radial oxygen loss (ROL) into the rhizosphere. Leaf glutamine synthetase activity (GSA) and nitrate (NO3‐) reductase activity (NRA) of Yangdao 6 were significantly higher than those of Nongken 57, while there was no significant difference in root NRA between the cultivars.

The nitrification activities were maximal in rhizosphere soil, followed by those in the bulk soil and the root surface for both cultivars. The rhizosphere nitrification activity, NO3‐ concentration and abundance of ammonia‐oxidizing bacteria (AOB) associated with Yangdao 6 were always higher than those of Nongken 57. Therefore, we conclude that the greater N uptake by Yangdao 6 when compared to Nongken 57 can be mainly attributed to the bigger capacity for nitrification in Yangdao 6.
Induction of Murine AA Amyloidosis by Various Homogeneous Amyloid Fibrils and Amyloid‐like Synthetic Peptides
Authors: Liu Y. , Cui D. , Hoshii Y. , Kawano H. , Une Y. , Gondo T. , Ishihara T.
Published: October 2007

We investigated amyloid‐enhancing factor (AEF) activity of amyloid fibrils extracted from amyloid‐laden livers of mice, cow, cheetah, cat and swan. All amyloid fibrils were confirmed to be amyloid protein A (AA) by an immunohistochemical analysis. We found that these fibrils accelerated the deposition of amyloid in an experimental mouse model of AA amyloidosis. Furthermore, the degree of deposition was dependent on the concentration of fibrils. When we compared the minimal concentration of amyloid fibrils needed to induce deposition, we found that these fibrils showed different efficiencies. Murine amyloid fibril induced amyloid deposition more efficiently than cow, cat, cheetah or swan amyloid fibrils. These data suggest that amyloid deposition is preferentially induced by amyloid fibrils with the same primary sequence as the endogenous amyloid protein. We then analysed the AEF activity of synthetic peptides, synthesized corresponding to amino acids 1–15 of mouse SAA (mSAA), 2–15 of cow SAA (bSAA), 1–15 of cat SAA (cSAA), which was the same as cheetah, and the common amino acids 33–45 of these four SAA (aSAA). We found that mSAA, bSAA and cSAA formed amyloid‐like fibrils in morphology and showed similar AEF properties to those of native amyloid fibrils. Although aSAA also formed highly ordered amyloid‐like fibrils, it showed weaker AEF activity than the other synthetic fibrils. Our results indicate that amyloidosis is transmissible between species under certain conditions; however, the efficiency of amyloid deposition is species‐specific and appears to be related to the primary amino acid sequence, especially the N‐terminal segment of the amyloid protein.
Generation of Anti‐tumour Effector T Cells from Naïve T Cells by Stimulation with Dendritic/tumour Fusion Cells
Authors: Ishida A. , Tanaka H. , Hiura T. , Miura S. , Watanabe S. , Matsuyama K. , Kuriyama H. , Tanaka J. , Kagamu H. , Gejyo F. , Yoshizawa H.
Published: October 2007

Tumour‐draining lymph node T cells are an excellent source of effector T cells that can be used in adoptive tumour immunotherapy because they have already been sensitized to tumour‐associated antigens in vivo. However, such tumour‐specific immune cells are not readily obtained from the host due to poor immunogenicity of tumours and reduced host immune responses. One obstacle in implementation of adoptive immunotherapy has been insufficient sensitization and expansion of tumour‐specific effector cells. In this study, we aim to improve adoptive immunotherapy by generating anti‐tumour effector T cells from naïve T lymphocytes. We attempted to achieve this by harnessing the advantages of dendritic cell (DC)‐based anti‐cancer vaccine strategies. Electrofusion was routinely employed to produce fusion cells with 30–40% efficiency by using the poorly immunogenic murine B16/F10 cell line, D5 cells, and DC generated from bone marrow cells. CD62L‐positive T cells from spleens of naïve mice and the fusion cells were cocultured with a low concentration of IL‐2. After 9 days of culture, the antigen‐specific T cells were identified with an upregulation of CD25 and CD69 expression and a downregulation of CD62L expression. These cells secreted IFN‐γ upon stimulation with irradiated tumour cells. Moreover, when transferred into mice with 3‐day established pulmonary metastases, these cells with coadministration of IL‐2 exhibited anti‐tumour efficacy. In contrast, naïve T cells cocultured with a mixture of unfused DC and irradiated tumour cells did not exhibit anti‐tumour efficacy. Our strategy provides the basis for a new approach in adoptive T cell immunotherapy for cancer.
A natural antisense transcript of the Petunia hybrida Sho gene suggests a role for an antisense mechanism in cytokinin regulation
Authors: Zubko Elena , Meyer Peter
Published: October 2007

The Sho gene from Petunia hybrida encodes an enzyme responsible for the synthesis of plant cytokinins. The 3′ region of the Sho gene contains a promoter in the opposite orientation that produces a partially overlapping antisense transcript. Although Sho expression varies significantly in individual cell types, the sense and antisense transcript levels maintain a stable ratio in most tissue types. In reporter lines for the antisense promoter, we observed a change in antisense promoter activity in newly formed tissue that had been induced by prolonged culture on cytokinins or following decapitation. We interpret these data as a reflection of tissue‐specific threshold levels for activation of the antisense transcript. In all tissue types tested, we detect a pool of antisense RNA of approximately 35 nt, which derives from the region where Sho sense and antisense transcripts overlap. We detect a second pool of putative dsRNA breakdown products of approximately 24 nt in all tissues tested, except roots, which are the main source of cytokinin synthesis. Our data suggest that antisense transcription can be activated in a tissue‐specific manner to adjust local cytokinin synthesis via degradation of Sho dsRNA. We therefore propose that, in addition to cytokinin transport and inactivation, regulation of local cytokinin synthesis via antisense transcription represents yet another device for the complex control of local cytokinin levels in plants.
Specialization of CDC27 function in the Arabidopsis thaliana anaphase‐promoting complex (APC/C)
Authors: Pérez‐Pérez José M. , Serralbo Olivier , Vanstraelen Marleen , González Cristina , Criqui Marie‐Claire , Genschik Pascal , Kondorosi Eva , Scheres Ben
Published: October 2007

To investigate the specialization of the two Arabidopsis CDC27 subunits in the anaphase‐promoting complex (APC/C), we analyzed novel alleles of HBT/CDC27B and CDC27A, and characterized the expression of complementing HOBBIT (HBT) protein fusions in plant meristems and during the cell cycle. In contrast to other APC/C mutants, which are gametophytic lethal, phenotypes of weak and null hbt alleles indicate a primary role in the control of post‐embryonic cell division and cell elongation, whereas cdc27a nulls are phenotypically indistinguishable from the wild type. However, cdc27a hbt double‐mutant gametes are non‐viable, indicating a redundant requirement for both CDC27 subunits during gametogenesis. Yeast‐two‐hybrid and pulldown studies with APC/C components suggest that the two Arabidopsis CDC27 subunits participate in several complexes that are differentially required during plant development. Loss‐of‐function analysis, as well as cyclin B reporter protein accumulation, indicates a conserved role for the plant APC/C in controlling mitotic progression and cell differentiation during the entire life cycle.
Original article: Impact of allergic rhinitis on asthma: effects on spirometric parameters
Authors: Ciprandi G. , Cirillo I. , Pistorio A.
Published: October 2007

Background:  Close association exists between allergic rhinitis and asthma. Moreover, allergic rhinitis is a strong risk factor for the onset of asthma in adults. This study was aimed at evaluating a large group of patients with moderate‐to‐severe and persistent allergic rhinitis alone for investigating the presence of spirometric abnormalities and possible risk factors related to it.

Methods:  A total of 392 patients with persistent allergic rhinitis were prospectively and consecutively evaluated. Clinical examination, skin prick test and spirometry were performed in all patients.

Results:  There were 24 (6.1%) patients with forced vital capacity (FVC < 80%) of predicted, 50 (12.8%) with forced expiratory volume in the first second (FEV1 < 80%) of predicted and 341 (87.0%) with forced expiratory flow at 25% and 75% of the pulmonary volume (FEF25–75) < 80% of predicted. The logistic regression analysis evidenced that rhinitis duration (ORAdj: 1.9/year) and sensitization to house dust mites (ORAdj: 8.2) were significantly associated with impaired values of 2 or 3 spirometric parameters.

Conclusion:  This study highlights the close link between upper and lower airways and the role of some risk factors, such as duration and mites sensitization, as early prognostic markers of bronchial involvement in patients with moderate‐to‐severe and persistent allergic rhinitis alone.
Identification of strain‐specific DNA of Actinobacillus actinomycetemcomitans by representational difference analysis
Authors: Mena J. , Chen C.
Published: October 2007

A genomic subtraction method, the representational difference analysis (RDA), was tested for its use in identifying strain‐specific DNA in Actinobacillus actinomycetemcomitans. Subtraction of strain D7S with strain HK1651 yielded D7S‐specific 2.3‐kilobase (kb) islet‐A and 5.3‐kb islet‐B. Islet‐A contains a 1.5‐kb region that is homologous to a region found in the A. actinomycetemcomitans plasmid pVT745. Islet‐B contains a 2.1‐kb homolog of vgr, a component of a DNA repeat element rhs. The distribution of these islets among A. actinomycetemcomitans strains was further examined by polymerase chain reaction. Islet‐A was found in nine serotype a and two serotype b strains but was missing from 34 strains. Islet‐B was found in one serotype a strain, four serotype d strains and two serotype e strains, but was missing from 34 strains.
Comparison of restriction enzyme analysis and amplified fragment length polymorphism typing of Porphyromonas gingivalis isolated from spouses
Authors: Rijnsburger M. C. , Delwel S. , Boutaga K. , Van Steenbergen T. J. M. , Savelkoul P. H. M. , Van Winkelhoff A. J.
Published: October 2007

Introduction:  In the past, theories on the transmission of Porphyromonas gingivalis between individuals have been based on, among other techniques, restriction enzyme analysis (REA) of bacterial DNA. Currently, amplified fragment length polymorphism (AFLP) may be a more sophisticated alternative. The possibility of automatic pattern analysis and digital storage of the typing data enables the comparison of patterns from a large number of strains in a broad time frame. The aim of this study was to compare REA profiles with AFLP patterns of P. gingivalis strains isolated from periodontitis patients and their spouses.

Methods: Forty‐two P. gingivalis strains were isolated from different sites in the mouth from six adult patients with periodontitis and their spouses. DNA of the bacterial isolates was subjected to REA and AFLP analysis.

Results:  One single type of P. gingivalis was found in each individual with both methods, regardless of the site of isolation. Indistinguishable types were found in four of the six couples with both techniques. Different types were found in two couples with both the REA and the AFLP method.

Conclusions:  The AFLP typing technique confirms earlier observations on the transmission of P. gingivalis between spouses. This new technique can replace REA typing.
Induction of immune responses and prevention of alveolar bone loss by intranasal administration of mice with Porphyromonas gingivalis fimbriae and recombinant cholera toxin B subunit
Authors: Takahashi Y. , Kumada H. , Hamada N. , Haishima Y. , Ozono S. , Isaka M. , Yasuda Y. , Tochikubo K. , Umemoto T.
Published: October 2007

Introduction:  Adult periodontitis is initiated by specific periodontal pathogens represented by Porphyromonas gingivalis; however, an effective measure for preventing the disease has not yet been established. In this study, the effectiveness of a vaccine composed of fimbriae of P. gingivalis and recombinant cholera toxin B subunit (rCTB) was evaluated using BALB/c mice.

Methods:  Fimbriae and rCTB were co‐administered intranasally to BALB/c mice on days 0, 14, 21, and 28. On day 35, mice were sacrificed to determine immunoglobulin levels in serum, saliva, and nasal and lung extracts by enzyme‐linked immunosorbent assay. The prevention effect of the vaccine on P. gingivalis‐induced periodontitis in mice was evaluated by measuring alveolar bone loss.

Results:  The rCTB significantly increased serum immunoglobulin (Ig)A levels when mice were administered with a minimal amount (0.5 μg) of the fimbrial antigen. The adjuvant effect on serum IgG production was indistinct because the minimal amount of the antigen still induced a large amount of IgG. In contrast to systemic responses, a fimbria‐specific secretory IgA response was strongly induced by co‐administration of rCTB and 0.5 μg fimbriae; the same amount of the antigen alone scarcely induced a response. Histopathological examination revealed IgA‐positive plasma cells in the nasal mucosal tissue but no observable mast cells in the area. In addition, nasal administration of the fimbrial vaccine significantly protected the mice from P. gingivalis‐mediated alveolar bone loss.

Conclusion:  Nasal vaccination with a combination of fimbriae and rCTB can be an effective means of preventing P. gingivalis‐mediated periodontitis.
Degradation of serine‐containing oligopeptides by Peptostreptococcus micros ATCC 33270
Authors: Uematsu H. , Hossain M. Z. , Alam T. , Ikeda T. , Kuvatanasuchati J. , Hoshino E.
Published: October 2007

Background/aims:  Microorganisms of Peptostreptococcus micros are asaccharolytic, anaerobic gram‐positive cocci that are frequently isolated from human oral sites such as periodontal pockets. Preliminary study showed that several amino acids, including serine, enhanced slightly the growth of P. micros. Therefore, we investigated the degradation of serine and serine‐containing oligopeptides.

Methods:  Metabolic end products were determined with high‐performance liquid chromatography. The related enzymatic activities in cell‐free extract were also assayed.

Results:  Washed P. micros degraded serine‐tripeptides (Ser‐Ser‐Ser), and produced formate, pyruvate, acetate, and ammonia. They also degraded serinyl‐tyrosine (Ser‐Tyr) to the same products. Related enzymatic activities, such as serine dehydratase, pyruvate formate‐lyase, formate dehydrogenase, pyruvate oxidoreductase, phosphate acetyltransferase, and acetate kinase, were detected in the cell‐free extract, indicating that the organisms produced ATP in the serine metabolism.

Conclusion:  P. micros utilized serine‐containing oligopeptides as exogenous metabolic substrates rather than serine itself, and degraded Ser‐Ser‐Ser and Ser‐Tyr to formate, pyruvate, acetate, and ammonia with ATP generation.
Evaluation of the microbiota of primary endodontic infections using checkerboard DNA–DNA hybridization
Authors: Sassone L. , Fidel R. , Figueiredo L. , Fidel S. , Faveri M. , Feres M.
Published: October 2007

Background/aims:  The aim of this study was to evaluate the composition of the microbiota of primary endodontic infections in 111 selected cases of single‐rooted teeth with necrotic pulp.

Methods:  Samples were collected from the root canals using #15 Hedströen‐type files and two sterile paper points, which were introduced 1 mm short of the apical foramen. The presence, levels, and proportions of 40 different bacterial species in each sample were determined using DNA probes and checkerboard DNA–DNA hybridization techniques.

Results:  The mean number of species per sample was 22. Enterococcus faecalis (89.3%), Campylobacter gracilis (89.3%), Leptotrichia buccalis (89.3%), Neisseria mucosa (87.5%), Prevotella melaninogenica (86.6%), Fusobacterium nucleatum ssp. vincentii (85.7%), Eubacterium saburreum (75.9%), Streptococcus anginosus (75%), and Veillonella parvula (74.1%) were the most prevalent species. The species found in highest mean counts (over 105) were F. nucleatum ssp. vincentii (13.14 × 105), E. saburreum (5.67 × 105), E. faecalis (5.38 × 105), N. mucosa (4.19 × 105), V. parvula (3.63 × 105), C. gracilis (3.46 × 105), Treponema socranskii (3.34 × 105), Porphyromonas endodontalis (2.96 × 105), Porphyromonas gingivalis (2.85 × 105), Micromonas micros (2.81 × 105), Prevotella nigrescens (2.68 × 105) and Fusobacterium nucleatum ssp. nucleatum (2.64 × 105). Most of these species were also found in high proportions.

Conclusions:  Our results suggest that several bacterial species considered to be oral pathogens seem to be implicated in the etiology of primary endodontic infections.
Effect of root canal procedures on endotoxins and endodontic pathogens
Authors: Vianna M. E. , Horz H.‐P. , Conrads G. , Zaia A. A. , Souza‐Filho F. J. , Gomes B. P. F. A.
Published: October 2007

Background/aims:  The purpose of this study was to determine the amount of endotoxin (lipopolysaccharide) and cultivable bacteria in human necrotic root canals before (S1) and after chemo‐mechanical preparation using chlorhexidine (CHX) gel as auxiliary chemical substance (S2), and after 7 days of intracanal dressing (S3) in order to evaluate the anti‐endotoxin and antimicrobial effects of endodontic procedures.

Method:  Twenty‐four teeth were selected for the present study. Chemo‐mechanical preparation was performed using 2% CHX gel and three different intracanal medicaments [Ca(OH)2 paste; 2% CHX gel; and Ca(OH)2 + 2% CHX gel]. A quantitative chromogenic Limulus amoebocyte lysate assay was used to measure the amount of endotoxin. Aerobic and anaerobic techniques were used to isolate and identify bacteria, and to determine the bacterial reduction by counting colony‐forming units (CFU).

Results:  Endotoxins and bacteria were present in 100% of the initial samples, with endotoxin concentration ranging from 62.93 to 214.56 UE/ml and CFU ranging from 4 × 105 to 2.6 × 106. After chemo‐mechanical preparation a mean endotoxin reduction of 44.4% was found. Eight (33.3%) root canals were still positive by culture analysis with a mean reduction of bacteria (CFU) of 99.96%. After 7 days of intracanal dressing, endotoxin concentration decreased by only 1.4% compared with S2, and residual bacteria were recovered by culture analysis in 13 cases (54.1%). No significant difference was found among different intracanal medicaments.

Conclusion:  Relatively high values of endotoxin were still present in the root canal after chemo‐mechanical preparation although the majority of bacteria were eliminated. No improvement was achieved by 7 days of intracanal dressing.
Improved accuracy in terminal restriction fragment length polymorphism phylogenetic analysis using a novel internal size standard definition
Authors: Takeshita T. , Nakano Y. , Yamashita Y.
Published: October 2007

Background:  Terminal restriction fragment length polymorphism (T‐RFLP) analysis is commonly used to analyze microbial communities, including oral microflora. However, accurate identification of terminal restriction fragment (T‐RF) origins is prevented by unpredictable errors in sizing, thus necessitating the clone library analysis. To minimize sizing errors, we proposed optimizing the size definition of internal standards.

Methods:  GeneScan‐1000 ROX was regenerated as an internal standard by redefining the fragment sizes in terms of molecular weight (MW) based on their mobility relative to 6‐carboxyfluorescein (FAM) ‐labeled restriction fragments derived from the 16S recombinant RNA gene of Porphyromonas gingivalis. Using the new size definition, the average sizing error among eight oral bacteria from six phyla was estimated and compared with that of the conventional method. Microbial communities isolated from saliva were analyzed using the new MW size definition. Bacterial species were assigned to peaks using TRFMA, a Web‐based tool for T‐RFLP analysis, and compared with those identified in a clone library analysis.

Results:  Using the new size definition, the average sizing error for 40 T‐RFs was drastically reduced from 2.42 to 0.62 bases, and large sizing errors (more than two bases) were eliminated. More than 90% of the total bacterial clones detected by the clone library analysis were assigned by T‐RFLP.

Conclusion:  The size definition of the newly constructed internal standards reduced fragment sizing errors and allowed for accurate assignment of bacteria to peaks by the T‐RFLP analysis. This provided a more effective means for studying microbial communities, including the oral microflora.
Long‐term engraftment following transplantation of pig pancreatic primordia into non‐immunosuppressed diabetic rhesus macaques
Authors: Rogers S.A. , Chen F. , Talcott M.R. , Faulkner C. , Thomas J.M. , Thevis M. , Hammerman M.R.
Published: October 2007

Abstract:  Background:  Transplantation therapy for human diabetes is limited by a shortage of donor organs, and transplant function diminished over time by cell death and limited potential for expansion of beta cells in pancreas or islets. Outcomes are complicated by immunosuppression. A way to overcome supply and expansion problems is to xenotransplant embryonic tissue. Previously, we have shown that beta cells originating from embryonic day (E) 28 (E28) pig pancreatic primordia transplanted into the mesentery of streptozotocin (STZ)‐diabetic (type 1) Lewis rats or Zucker Diabetic Fatty (ZDF) diabetic (type 2) rats engraft and normalize glucose tolerance without the need for host immune‐suppression.

Methods:  In this study, we transplant E28 pig pancreatic primordia in the mesentery of STZ‐diabetic rhesus macaques.

Results:  Long‐term engraftment of pig beta cells within liver, pancreas and mesenteric lymph nodes post‐transplantation of E28 pig pancreatic primordia into STZ‐diabetic rhesus macaques is demonstrated by electron microscopy, positive immune‐histochemistry for insulin, and positive RT‐PCR and in situ hybridization for porcine proinsulin mRNA. Insulin requirements were reduced in one macaque followed over 22 months post‐transplantation and porcine insulin detected in plasma using sequential affinity chromatography, HPLC and mass spectrometry. Of potential importance for application of this transplantation technology to treatment of diabetes in humans and confirmatory of our previous findings in Lewis and ZDF rats, no host immunosuppression is required.

Conclusions:  Under selected circumstances, pancreatic primordia elicit a muted immune response relative to more differentiated tissue, such that engraftment occurs in non‐immunosuppressed hosts. Our findings that pig pancreatic primordia engraft long‐term in non‐immunosuppressed STZ‐diabetic rhesus macaques establishes the potential for their use in human diabetics.
Cranial Features and Karyotypes of Two Hedgehogs (Insectivora: Erinaceidae) from Iran
Authors: Karataş A. , Mouradi Gharkheloo M. , Kankiliç T.
Published: October 2007

Karyological features of Erinaceus concolor and Hemiechinus auritus were studied from Zenjan (North Iran). The diploid number of chromosomes (2n), the total numbers of chromosomal arms (NF) and the numbers of autosomal arms (NFa) were determined as 2n = 48, NF = 92, NFa = 88 for E. concolor and as 2n = 50, NF = 98, NFa = 94 for H. auritus. Although the same dental formulae were established as 3.1.3.3/2.1.2.3 = 36 for both of species, they can be identified with some dental and cranial features from each other. Additionally, phallus of E. concolor was described.
Histology and Ultrastructure of the Developing Superficial Digital Flexor Tendon in Rabbits
Authors: Oryan A. , Shoushtari A. H.
Published: October 2007

This study was designed to investigate the developmental changes in the superficial digital flexor tendon (SDFT) of the white New Zealand rabbit, from 5 to 7 days pre‐natal to 112 days post‐natal (PN), at histological and ultrastructural levels. The tendons changed from being highly cellular with 13079 ± 2538 cell/mm2 and little directional organization to a longitudinally oriented, predominantly connective tissue structure with relatively few, mature tenocytes (2384 ± 365 cell/mm2). Fibrillogenesis was seen in isolated vacuoles of fibroblast cytoplasm during fetal life as well as after birth, but less frequently with increasing age. At the ultrastructural level, there was a progressive increase in the mean diameter of collagen fibrils with age throughout the population, until PN day 28. A bimodal distribution of collagen fibril size was first observed on PN day 56 while at day 112, the fibrils were fully differentiated and showed a multimodal size distribution. The development of elastic fibres preceded that of collagen fibres and accompanied progressively more marked sinusoidal crimping in collagen fibres on PN days 7 and 14. The tendons of older animals became less tightly crimped. The cells of the epi‐ and endotenon were less mature and relatively undifferentiated compared with the cells in the body of the tendon of the same age, which might explain their ability to initiate healing of tendon injuries in older animals. In conclusion, extensive changes were observed in tendons during growth and maturation, with diameters of collagen fibrils, tissue orientation and cellularity being the parameters affected, probably due largely to increased physical activity.
The Development of the Digestive Tract in Larval European Catfish (Silurus glanis L.)
Authors: Kozarić Z. , Kužir S. , Petrinec Z. , Gjurčević E. , Božić M.
Published: October 2007

The European catfish, Silurus glanis L., has become an important aqua cultural fish in Croatia, and it is cultivated extensively in ponds in polyculture with carps. The development of the digestive tract in S. glanis was studied with the aim of improving intensive fish production. Research was carried out on S. glanis larval stadium from 1‐ to 19‐day post‐hatching (DPH). The main histological methods used were: haematoxylin and eosin staining, periodic acid Schiff staining (PAS), Alcian blue (AB) and toluidin blue staining (TB). A yolk sac was present during the first 5 days (1–5‐DPH). During the initial 3‐DPH period, there was no trace PAS and AB activity in the digestive tract. Differentiation of the digestive tract began at 3‐ to 5‐DPH. The oesophagus was positive for AB at 5‐DPH, PAS and TB after 7‐DPH. Differentiation of enterocytes began at 5‐DPG and the intestines were complete at 11‐DPH. Development of liver and pancreas was also studied. The analysis of data obtained in this study suggests that after 5‐DPH catfish larvae have morphologically completed digestive tracts.
Habituation by a tortoise beetle to the deterrents of spinach
Authors: Nagasawa A. , Matsuda K.
Published: October 2007

Abstract:  Feeding responses of Cassida nebulosa to spinach vary depending on prior dietary experience. Adults reared on spinach, eat much spinach whereas those reared on Chenopodium album var. centrorubrum rarely eat it. However, there are a few reports that address the mechanisms of this phenomenon, which is termed ‘induction of preference’. It has been presumed that this phenomenon arises in response to a reduction of sensitivity to the feeding deterrents contained in spinach as a result of early experience feeding on it. Therefore, we attempted to verify the effects of the feeding deterrent that was isolated from spinach. The feeding‐deterrent response was reduced in adults reared on spinach compared with adults reared on C. album var. centrorubrum. Additionally, consumption of spinach by adults reared on deterrent‐treated leaves of C. album var. centrorubrum was significantly higher than that by adults reared on control leaves of C. album var. centrorubrum. These results indicate that C. nebulosa acquires the ability to tolerate spinach because sensitivity to the feeding deterrent contained in spinach is reduced by early experience with the deterrent.
Genetic diversity among pearl millet maintainers using microsatellite markers
Authors: Kapila R. K. , Yadav R. S. , Plaha P. , Rai K. N. , Yadav O. P. , Hash C. T. , Howarth C. J.
Published: October 2007

Genetic diversity among 70 maintainers and two pollinators of sub‐Saharan and Indian origin was studied for simple sequence repeat (SSR) loci using 34 primer pairs. A total of 213 alleles were detected with an average of 6.26 alleles per locus. Polymorphic information content (PIC) ranged from 0.05 to 0.96 with a mean of 0.58 for the SSR loci. Mean PIC across the linkage groups and number of alleles in dinucleotide motifs varied significantly. The lowest PIC (0.239) for linkage group 6 indicated comparatively conserved nature of this linkage group. Genetic similarity estimates ranged from 0.05 to 0.73 with an average value of 0.29. This indicated sufficient diversity among the maintainer and pollinator lines. The 72 lines fell in five clusters, and the clustering pattern corroborated with their pedigree and characteristic traits. Pollinator ICMR 356 was more diverse from the maintainer lines analysed, and can be a potential parent for pearl millet hybrid development.
A case of widespread non‐pigmented hair regrowth in diffuse alopecia areata
Authors: Dinh Quan Q , Chong Alvin H
Published: October 2007

A 62‐year‐old woman presented with diffuse alopecia areata affecting more than 50% of her scalp hairs with sparing of non‐pigmented hairs. She was treated with a 2‐month course of oral prednisolone and extensive hair regrowth occurred. The new hairs were completely non‐pigmented. As a result, the patient's hair colour is now strikingly and completely white, with a normal hair density. This response has been sustained for 6 months after cessation of treatment.
Mast cells and vascular density have prognostic value in Merkel cell carcinoma
Authors: Ng Lydia , Beer Trevor W.
Published: October 2007

Aims  Merkel cell carcinoma (MCC) is said to be the most aggressive of all skin cancers, with frequent metastases and a significant mortality rate. This study examined the role of mast cells and vascular density in the prognosis of Merkel cell carcinoma.

Methods  Mast cells and vessels were stained immunohistochemically in 36 MCC with antibodies to tryptase and CD34 respectively. The number of stainable mast cells and vessels were quantified within the tumours and surrounding stroma. Other clinical and histological parameters were also examined, statistically analysed and compared to subsequent clinical course and prognosis.

Results  Prognosis was poorer with higher mast cell numbers (p < 0.002) and with increased vascular density (p = 0.005). A one unit increase in mast cells predicted a 1.7 times increased risk of death. Prognosis was also adversely affected by the presence of lymphovascular invasion (p = 0.03) and larger tumour size (p = 0.05).

Discussion  Although factors predicting the behaviour of MCC are largely unknown, various parameters have been proposed as potential prognostic markers [Llombart, 2005].

Mast cells are pivotal in immunity, but a prognostic role for mast cells in malignancies has also been suggested, possibly through mitogenesis and angiogenesis [Ch'ng, 2006]. Vascular density, as a surrogate for angiogenesis, may have prognostic value in certain cancers, e.g. melanoma [Depasquale, 2005].

In this study, increased mast cells counts and vascular density were associated with an adverse prognosis in MCC. Evaluation of mast cell infiltrates and vascular density can provide prognostic data and may ultimately help to guide appropriate treatment selection for this aggressive form of skin cancer.
ANTIPROLIFERATIVE ACTIVITY OF PAEONIFLORIN IS THROUGH CELL CYCLE ARREST AND THE FAS/FAS LIGAND‐MEDIATED APOPTOTIC PATHWAY IN HUMAN NON‐SMALL CELL LUNG CANCER A549 CELLS
Authors: Hung Jen‐Yu , Yang Chih‐Jen , Tsai Ying‐Ming , Huang Hurng‐Wern , Huang Ming‐Shyan
Published: October 2007

Paeoniflorin (PF), isolated from the paeony root, is reported to have immunoregulatory, neuromuscular blocking, anticonvulsant, antihyperglycaemic and antihypotensive effects.

The present study investigated the antiproliferative activity of PF. The results showed that PF inhibited the proliferation of A549 by blocking cell cycle progression in the G0/G1 phase and inducing apoptosis.

An ELISA showed that G0/G1 phase arrest may be due to p53‐independent induction of p21/wild‐type p53‐activated fragment 1 (WAF1). Increased protein expression of Fas/apoptosis‐1 (APO‐1) and its two ligands, membrane‐bound Fas ligand and soluble Fas ligand, may be responsible for the PF‐induced apoptosis.

This is the first study to show that the induction of p21/WAF1 and the activity of the Fas/Fas ligand apoptotic system may participate in the antiproliferative activity of PF in A549 cells.
PROTECTION OF ISCHAEMIC‐REPERFUSED RAT HEART BY DIMETHYLAMILORIDE IS ASSOCIATED WITH INHIBITION OF MITOCHONDRIAL PERMEABILITY TRANSITION
Authors: Prendes María G Marina , Torresín Emilia , González Marcela , Fernández María A , Perazzo Juan C , Savino Enrique A , Varela Alicia
Published: October 2007

The aim of the present study was to assess whether protection afforded by the Na+/H+ exchanger blocker dimethylamiloride (DMA) is associated with inhibition of mitochondrial permeability transition (MPT). The effects of DMA were compared with those of cyclosporine (Cs) A, an inhibitor of MPT.

Rat hearts were Langendorff perfused with Krebs’–bicarbonate medium containing 10 mmol/L glucose and were subjected to 25 min no‐flow global ischaemia and 30 min reperfusion in the presence or absence of 10 µmol/L DMA or 0.2 µmol/L CsA. Cell viability was measured using tetrazolium stain. The MPT was determined by loading hearts with 2‐deoxy‐[3H]‐glucose (2DG), which enters mitochondria only during MPT. Total heart 2DG content as an estimation of the extent of tissue damage was also measured. To assess whether DMA has any direct effect on glycolysis, a cell‐free heart extract containing all the glycolytic enzymes was used.

Dimethylamiloride improved functional recovery (rate–pressure product) from 24 ± 7 to 68 ± 11% (P < 0.01) at reperfusion end, attenuated the increase in left ventricular end‐diastolic pressure (from 29 ± 7 to 6 ± 3% 10 min after reperfusion onset; P < 0.01), improved cell viability (from 21.2 ± 6.6 to 69.6 ± 7.1% at reperfusion end; P < 0.05) and lessened lactate accumulation at the end of ischaemia (119 ± 15 vs 163 ± 14 µmol/g dry weight; P < 0.05). Dimethylamiloride limited MPT : 2DG mitochondrial entrapment, being 33.1 ± 14.2 and 96.3 ± 14.0 at reperfusion end in the treated and control hearts, respectively (P < 0.05), and concomitantly raised total 2DG content (51.3 ± 4.4 vs 86.8 ± 1.7 × 103 d.p.m./g wet weight in control and treated groups, respectively; P < 0.05). Cyclosporine A improved functional recovery and attenuated the amplitude of ventricular diastolic pressure in ischaemic–reperfused hearts. It also reduced mitochondrial entrapment (67.3 ± 7.7%; P < 0.05 vs control) and increased total cell 2DG content (162.3 ± 1.3 × 103 d.p.m./g wet weight; P < 0.01 vs control) at the end of reperfusion. Dimethylamiloride did not affect glucose consumption and lactate production in the cell‐free heart extract.

In conclusion, DMA protects against the noxious effects of ischaemia–reperfusion and inhibits MPT, coinciding with present and previous findings concerning the effects of CsA. Dimethylamiloride also diminished lactate accumulation, although it did not exhibit any direct effect on glycolysis. These data suggest that blockade of Na+/H+ exchange by DMA attenuates the extent of MPT in ischaemic–reperfused rat heart.