bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of C‐reactive protein and pentosan polysulphate on human complement activation
Authors: Klegeris AndIs , Singh Edith A. , MCGeer Patrick L.
Published: March 2003

Complement (C) activation is believed to play an adverse role in several chronic degenerative disease processes, including atherosclerosis, myocardial infarction and Alzheimer's disease. We developed several in vitro quantitative assays to evaluate processes which activate C in human serum, and to assess candidates which might block that activation. Binding of C‐reactive protein (CRP) to immobilized cell surfaces was used as a tissue‐based method of activation, while immunoglobulin G in solution was used as a surrogate antibody method. Activation was assessed by deposition of C fragments on fixed cell surfaces, or by capture of C5b‐9 from solution. We observed that several cell lines, including SH‐SY5Y, U‐937, THP‐1 and ECV304, bound CRP and activated C following attachment of cells to a plastic surface by means of air drying. Treatment of human neuroblastoma SH‐SY5Y cells with the reactive oxygen intermediates generated by xanthine (Xa) – xanthine oxidase (XaOx) prior to air drying or by hydrogen peroxide solutions after air drying, enhanced C activation, possibly through oxidation of the cell lipid membrane. Several C inhibitors were tested for their effectiveness in blocking these systems. Pentosan polysulphate (PPS), an orally active agent, blocked C activation in the same concentration range of 1–1000 µg/ml as heparin, dextran sulphate, compstatin and fucoidan. PPS may have practical application as a C inhibitor.
Tumour necrosis factor‐α is a fundamental cytokine in autoimmune thyroid disease induced by thyroglobulin and lipopolysaccharide in interleukin‐12 p40 deficient C57BL/6 mice
Authors: Zaccone Paola , Fehérvári Zoltán , Cooke Anne
Published: March 2003

Experimental autoimmune thyroiditis (EAT) is inducible in mice by immunization with mouse thyroglobulin (mTg) together with adjuvant, either lipopolysaccharide (LPS) or complete Freund's adjuvant (CFA). The severity of the disease is dependent on the mouse strain and on the adjuvant used. We have previously shown that interleukin (IL)‐12 deficient C57BL/6 mice immunized with mTg and CFA develop a significantly less severe thyroid infiltration in comparison to wild type C57 BL/6 mice. This result indicated a pivotal role for IL‐12 in the development of thyroiditis induced with CFA and mTg. In the present study we demonstrate that IL‐12 deficiency does not impair EAT induction when LPS is used as adjuvant. We also demonstrate that peritoneal exudate cells from IL‐12‐deficient mice stimulated in vitro either with LPS or IL‐18 secrete high levels of tumour necrosis factor (TNF). Together the results emphasize the difference between the use of CFA and LPS in the induction of EAT, the importance of TNF‐α for the pathogenesis of LPS‐induced EAT, and also show the capacity of IL‐12‐deficient mice to develop a competent response to LPS.
Differential response of bovine monocyte‐derived macrophages and dendritic cells to infection with Salmonella typhimurium in a low‐dose model in vitro
Authors: Norimatsu Mari , Harris James , Chance Victoria , Dougan Gordon , Howard Christopher J. , Villarreal‐Ramos Bernardo
Published: March 2003

Exposing bovine dendritic cells (DC) and macrophages (MΦ) to Salmonella typhimurium at a ratio of 1 cell to 10 bacteria had a cytotoxic effect that was not evident with a ratio of 1000 cells to 1 bacterium. This lower dose was considered to mimic more closely the in vivo situation and a comparison was made with this model of the consequences of infection for MΦ and DC. DC infected with S. typhimurium up‐regulated cell surface expression of major histocompatibility class I (MHC‐I), MHC‐II, CD40, CD80 and CD86. In contrast, infected MΦ did not exhibit detectable changes in expression of cell surface molecules, except for a marginal increase in CD40. mRNA transcription for tumour necrosis factor‐α, interleukin (IL)‐1β, IL‐6 and inducible nitric oxide synthase was up‐regulated in both infected DC and infected MΦ, although mRNA transcription for granulocyte–macrophage colony‐stimulating factor and IL‐12p40 was up‐regulated only in infected DC and for IL‐10 was only in infected MΦ. Infected DC had an increased ability to stimulate both allogeneic and antigen‐specific T‐cell responses compared to non‐infected controls. In contrast, infected MΦ showed an increased ability to induce allogeneic responses but this was less than seen for DC and no enhancement of ability to induce antigen‐specific T cell responses was seen. Thus, in a low‐dose infection model that does not result in the cytotoxicity of a substantial percentage of antigen presenting cells, bovine MΦ and DC respond differently to infection with S. typhimurium and this could have important implications for the development of the immune response.
Immunomodulatory effects of cyclosporin A on human peripheral blood dendritic cell subsets
Authors: Tajima Kenichirou , Amakawa Ryuichi , Ito Tomoki , Miyaji Michihiko , Takebayashi Masashi , Fukuhara Shirou
Published: March 2003

Cyclosporin A (CsA) is a potent immuno‐suppressant and is approved for the treatment of various disease conditions. The molecular biological mechanism of CsA has been investigated intensively in T cells and has been shown to involve the intracellular calcineurin pathway. Recently, it was reported that CsA has capacities to affect not only T cells but also antigen‐presenting cells such as B cells and dendritic cells (DCs). DCs are a master regulator of immune responses that have an integral capacity to prime naive T cells. In the present study, we investigated the biological effects of CsA on human peripheral blood DC subsets: CD11c+ myeloid and CD11c− lymphoid subsets. CsA inhibited the up‐regulation of co‐stimulatory molecules induced with or without microbial stimuli and CD40L on both CD11c+ and CD11c− subsets. In addition, CsA negatively regulated the endocytic activity of CD11c+ DC during the immature state. CsA inhibited the interleukin‐12 (IL‐12) production, but augmented the IL‐10 production from the LPS‐stimulated CD11c+ subset, whereas CsA reduced the interferon‐α (IFN‐α) production from the CD11c− subset infected with Sendai virus (SV). Both the LPS‐stimulated CD11c+ subset and SV‐infected CD11c− subset preferentially induced the development of IFN‐γ‐producing T helper‐type 1 (Th1) cells. Pretreatment of these DC subsets with CsA inhibited the Th1 skewing. These findings suggested a DC‐mediated mechanism of immunosupression by CsA.
The effect of graphic style on data model interpretation
Authors: Nordbotten J. C. , Crosby M. E.
Published: March 2003

Abstract. Graphic data models are commonly used as a tool for presentation of information structures in the design, implementation, use and maintenance of the databases that support information systems. The methods proposed for database design assume that the use of graphic data models will enhance understanding of system specifications by both the end‐users and the implementers of the system. For this assumption to hold, the information presented in the graphic data model must be readily comprehensible so that the design, represented by the model, can be confirmed and implemented correctly. The lack of standard representations for graphic models has led to a variety of graphic styles. To date, there has been little focus on studying the effect graphic style has on model comprehension. We have studied the effect of three graphic styles proposed for data models on model legibility and interpretation. Our study shows a significant variation in model interpretation that can be attributed to the graphic syntax used. Graphic style appears to influence which model elements are included in the interpretation, as well as the way data models are read.
Low substitution rates in mitochondrial DNA in Mediterranean carabid beetles
Authors: Prüser F. , Mossakowski D.
Published: March 2003

The estimation of DNA substitution rates requires calibration with the geological events which caused the separation of populations. We used the disintegration of the landbridges between Morocco and Gibraltar as well as the Balearics and Spain at the end of the Messinian event (5.3 myr) in order to calibrate the DNA sequence data of the NADH‐dehydrogenase subunit 1 (ND1) in West Mediterranean Carabus species. The rates of molecular evolution we found are much lower than those reported in the literature and also vary from species to species. Therefore, in insects the calculation of divergence periods using a constant 2% substitution rate per myr is questionable.
Isolation of seven unique biogenic amine receptor clones from the honey bee by library scanning
Authors: Ebert P. R. , Rowland J. E. , Toma D. P.
Published: March 2003

The biogenic amine receptor genes constitute an ancient and highly divergent family within the larger superfamily of G‐protein‐coupled receptors. These receptors play a central role in modulating nerve cell activity and thus behaviour. Because the honey bee offers numerous advantages for behavioural studies we endeavoured to isolate as many members of this gene family as possible from the bee. We compared numerous approaches to gene isolation and found that PCR amplification from small subfractions of cDNA or genomic DNA libraries enabled us to isolate clones that are otherwise undetectable. In total we isolated seven biogenic amine receptor clones and identified five additional related sequences by low‐stringency Southern hybridization. Two clones, AmBAR4 and AmBAR6, are 84% and 72% identical to the Drosophila 5‐HT2 and D1b receptors, respectively, and probably represent orthologous genes. Phylogenetic analysis indicates that AmBAR5 clusters loosely with a variety of tyramine and octopamine receptors with which it shares <66% identity. The other four clones, AmBAR1, AmBAR2, AmBAR3 and AmBAR7, are weakly to moderately related (28–45% identical) to Drosophila dopaminergic or mammalian adrenergic receptors and probably represent receptors of these classes whose orthologues have not previously been isolated from any insect. The honey bee clones expand the size of the known insect biogenic amine receptor gene family to sixteen members. Therefore the size of the biogenic amine receptor gene family of insects approaches that of vertebrates. This is true despite the reduced behavioural and genetic complexity of the insects relative to vertebrate animals.
Lack of nucleotide variability in a beetle pest with extreme inbreeding
Authors: Andreev D. , Breilid H. , Kirkendall L. , Brun L. O. , Ffrench‐Constant R. H.
Published: March 2003

The coffee berry borer beetle Hypothenemus hampei (Ferrari) (Curculionidae: Scolytinae) is the major insect pest of coffee and has spread to most of the coffee‐growing countries of the world. This beetle also displays an unusual life cycle, with regular sibling mating. This regular inbreeding and the population bottlenecks occurring on colonization of new regions should lead to low levels of genetic diversity. We were therefore interested in determining the level of nucleotide variation in nuclear and mitochondrial genomes of this beetle worldwide. Here we show that two nuclear loci (Resistance to dieldrin and ITS2) are completely invariant, whereas some variability is maintained at a mitochondrial locus (COI), probably corresponding to a higher mutation rate in the mitochondrial genome. Phylogenetic analysis of the mitochondrial data shows only two clades of beetle haplotypes outside of Kenya, the proposed origin of the species. These data confirm that inbreeding greatly reduces nucleotide variation and suggest the recent global spread of only two inbreeding lines of this bark beetle.
Cloning and characterization of a cDNA encoding a male‐specific serum protein of the Mediterranean fruit fly, Ceratitis capitata, with sequence similarity to odourant–binding proteins
Authors: Thymlanou S. , Mavroidis M. , Kokolakis G. , Komitopoulou K. , Zacharopoulou A. , Mintzas A. C.
Published: March 2003

Male‐specific serum proteins (MSSPs) are low molecular weight proteins which accumulate in high amounts in the haemolymph of adult males of the medfly Ceratitis capitata. By screening an expression library with anti‐MSSP antibodies, we have isolated and determined the nucleotide sequence of a cDNA clone coding for one of the male‐specific polypeptides (MSSP‐α). The MSSP‐α mRNA encodes a polypeptide of 144 amino acids with a secretory signal sequence of sixteen amino acids. Southern analysis indicated that there are multiple copies of MSSP genes in the medfly genome. Northern analysis showed that the MSSP mRNAs are synthesized only in adult males. The accumulation pattern of these mRNAs during development suggests that the expression of the MSSP genes is developmentally regulated at both transcriptional and translational levels. The predicted peptide sequence of MSSP‐α shows significant similarity to a group of pheromone‐ and general odourant‐binding proteins of insects.
Invasion of one insect species, Adalia bipunctata, by two different male‐killing bacteria
Authors: Hurst G. D. D. , von der Schulenburg J. H. Graf , Majerus T. M. O. , Bertrand D. , Zakharov I. A. , Baungaard J. , Völkl W. , Stouthamer R. , Majerus M. E. N.
Published: March 2003

Male‐killing bacteria, which are inherited through the female line and kill male progeny only, are known from five different orders of insect. Our knowledge of the incidence of these elements has stemmed from discovery of their phenotype in different species. Our estimate of the frequency with which insects have been invaded by these elements therefore depends on each observation of the male‐killing phenotype within a species being associated with a single micro‐organism. We here record an example of a single insect species being infected with two taxonomically distinct male‐killing bacteria. Western European populations of the two‐spot ladybird, Adalia bipunctata, have previously been shown to bear a male‐killing Rickettsia. However, we here show that the majority of the male‐killing lines tested from Central and Eastern Europe do not bear this bacterium. Rather, 16S rDNA sequence analysis suggests male‐killing is associated with a member of the genus Spiroplasma. We discuss this conclusion in relation to the evolutionary genetics of male‐killing bacteria, and the evolution of male‐killing behaviour in the eubacteria.
Immunity proteins from mosquito cell lines include three defensin A isoforms from Aedesaegypti and a defensin D from Aedesalbopictus
Authors: Gao Y. , Hernandez V. P. , Fallon A. M.
Published: March 2003

An Aedes aegypti mosquito cell line, Aag‐2, exhibits a response to immune stimulation that is qualitatively similar to that of C7–10 cultured cells from the related mosquito, Aedes albopictus. Using SDS polyacrylamide gels, we found that a small peptide was preferentially induced by the treatment of growing cells with heat‐killed, Gram‐positive bacteria. By an analogy with other studies, this small peptide was postulated to be a member of the defensin family of insect immunity peptides. A differential display was used to obtain partial polymerase chain reaction products corresponding to mRNAs that were preferentially expressed in induced cells. One of these products, which contained the partial sequence of a defensin gene, was used to screen cDNA libraries from Ae. aegypti and Ae. albopictus cells. From Ae. aegypti cells, we found two previously described isoforms (A1 and A4) of mosquito defensin A, as well as a new isoform which we defined as A5. From Ae. albopictus cells, we found a new mature mosquito defensin, named D, which contains proline and isoleucine as the final amino acids. In both Ae. aegypti and Ae. albopictus cell lines, the expression of defensin mRNA was visible on Northern blots as early as 3 h after exposure to heat‐killed bacteria, and defensin mRNA abundance was maximal at 12–36 h after induction.
Phylogenetic relationships among Muscoidea (Diptera: Calyptratae) based on mitochondrial DNA sequences
Authors: Bernasconi M. V. , Valsangiacomo C. , Piffaretti J.‐C. , Ward P. I.
Published: March 2003

The utility of a mitochondrial DNA (mtDNA) fragment of about 1100 bp (including partial COI and COII sequences and tRNALeu) for evolutionary studies in Muscoidea is discussed. The species investigated are Scathophaga stercoraria, Microprosopa pallidicauda and Trichopalpus fraterna (family Scathophagidae), Musca domestica (Muscidae), Lasiomma seminitidum (Anthomyiidae) and Fannia armata (Fanniidae). Comparisons were made with published mtDNA sequences of Drosophila, Anopheles and three Calliphoridae species. The molecular phylogeny obtained here matches the classical morphological taxonomy reasonably well. This varies considerably, however, at different taxonomical levels. At a high taxonomic level, there is a clear separation between the Nematocera and the Brachycera, but the Calyptratae–Acalyptratae division is not always supported. At a lower taxonomic level, all species belonging to the same family are well grouped, but at an intermediate level, within the Calyptratae, it is impossible to clearly separate the Muscoidea and Calliphoridae, preventing a firm conclusion on the phylogenetic relationships among Muscoidea families. The entire COI sequence of S. stercoraria, as well as other mtDNA sequences (including the proximal portions of the COI gene, tRNATrp, tRNACys and tRNATyr genes) in Muscoidea species, are also presented and discussed.
Molecular identification and phylogeny of East African Simulium damnosum s.l. and their relationship with West African species of the complex (Diptera: Simuliidae)
Authors: Krüger A. , Gelhaus A. , Garms R.
Published: March 2003

The phylogenetic relationships of East and West African members of the Simulium damnosum complex were studied by sequence analyses of the mitochondrial 16s ribosomal RNA (rRNA) gene. Results suggest that: (i) the S. damnosum complex is divided into an East and a West African clade, and (ii) S. pandanophilum and the cytoform ‘Kiwira’ form an East African subbranch distinct from the ‘Sanje’ group. In contrast to former assumptions from cytogenetic analyses, our molecular data do not support a direct relationship between the East African S. kilibanum and the West African S. squamosum. Length differences of the rDNA internal transcribed spacer 1 (ITS‐1) turned out to be useful to distinguish between cytoforms.
Evidence for mitochondrial introgression between Anopheles bwambae and Anopheles gambiae
Authors: Thelwell N. J. , Huisman R. A. , Harbach R. E. , Butlin R. K.
Published: March 2003

There is evidence for introgression between the malaria vectors Anopheles gambiae and Anopheles arabiensis, which belong to the Anopheles gambiae complex. Here we consider their relationship to another member of the complex, Anopheles bwambae. Species identifications were made using rDNA‐PCR and revealed one An. bwambae/gambiae hybrid among 459 wild‐caught mosquitoes. A 237‐nucleotide region of the mitochondrial ND5 gene was sequenced in sixty‐nine individuals. The results suggest that there is an An. bwambae specific group of haplotypes. However, two An. bwambae individuals carried haplotypes typical of An. gambiae and An. arabiensis. This is the first evidence for introgression between An. bwambae and other species of the An. gambiae complex and suggests that introgression may be more widespread, also occurring between other members of the complex.
Comparison of esterase gene amplification, gene expression and esterase activity in insecticide susceptible and resistant strains of the brown planthopper, Nilaparvata lugens (Stål)
Authors: Vontas J. G. , Small G. J. , Hemingway J.
Published: March 2003

Organophosphorus and carbamate insecticide resistance in Nilaparvata lugens is based on amplification of a carboxylesterase gene, Nl‐EST1. An identical gene occurs in susceptible insects. Quantitative real‐time PCR was used to demonstrate that Nl‐EST1 is amplified 3–7‐fold in the genome of resistant compared to susceptible planthoppers. Expression levels were similar to amplification levels, with 1–15‐fold more Nl‐EST1 mRNA in individual insects and 5–11‐fold more Nl‐EST1 mRNA in mass whole body homogenates of resistant females compared to susceptibles. These values corresponded to an 8–10‐fold increase in esterase activity in the head and thorax of individual resistant insects. Although amplification, expression and activity levels of Nl‐EST1 in resistant N. lugens were similar, the correlation between esterase activity and Nl‐EST1 mRNA levels in resistant individuals was not linear.
Attempts to molecularly distinguish cryptic taxa in Anopheles gambiae s.s.
Authors: Gentile G. , Slotman M. , Ketmaier V. , Powell J. R. , Caccone A.
Published: March 2003

Analyses of inversions in polytene chromosomes indicate that, in West Africa, Anopheles gambiae (sensu stricto) may be a complex of more than a single taxonomic unit, and these units have been called chromosomal forms. In order to determine whether this genetic discontinuity extends to the rest of the genome, as would be expected if reproductive isolation exists, we have sequenced several regions of both the nuclear and mitochondrial genomes. With one exception, we were unable to identify any nucleotide sites that differentiate the chromosomal forms. The exception was the internal transcribed spacer (ITS) of the ribosomal DNA (rDNA). Three sites in this region distinguish Mopti chromosomal form from Savanna and Bamako in Mali and Burkina Faso. However, outside these two countries, the association between chromosomal form and rDNA type does not always hold. Together with the variants in the rDNA intergenic spacer (IGS) described in the accompanying papers (della Torre et al., 2001; Favia et al., 2001), we can recognize two major types of rDNA, Type I and Type II (corresponding to molecular forms S and M in della Torre et al., 2001). Type I is widespread in West Africa and is the only type found outside of West Africa (i.e. Tanzania and Madagascar). Type II is confined to West Africa. We were unable to detect any heterozygosity for the ITS types even in five collections containing both types. A sample from the island of São Tomé could not be classified into either Type I or Type II as the rDNA had characteristics of both. In general, our results confirm that An. gambiae is not a single pan‐mictic unit, but exactly how to define any new taxa remains problematic. Finally, we have found minor variants of the major rDNA types fixed in local populations; contrary to most previous studies, this suggests restricted gene flow among populations of this species.
Elongation Factor 1 α resolves the monophyly of the haplodiploid ambrosia beetles Xyleborini (Coleoptera: Curculionidae)
Authors: Jordal B. H.
Published: March 2003

Elongation Factor 1‐α was used to test the monophyly of the wood boring beetle tribe Xyleborini, where all species are haplodiploid and perform regular inbreeding by brother–sister mating. Due to their feeding requirements, being highly dependent on ophiostomatoid fungi which they cultivate in wood tunnels, monophyly may be expected due to nutritional constraints. During the course of analyses, two copies of EF‐1α were amplified in these beetles, differing in intron structure. The high similarity between paralogous amino acid sequences (93–94%) indicates a rather recent duplication in beetles, but phylogenetic analyses of different copies in insects rejected this hypothesis. Subsequent phylogenetic analyses of eighty orthologous sequences from Xyleborini and allied taxa, using the single‐intron bearing copy, were greatly improved in resolution and node support by including the intron sequences (c. 60 bp). Most analyses resulted in a monophyletic Xyleborini, implying one origin of fungus feeding in this tribe. However, clear evidence for a polyphyletic Xyleborus and three more xyleborine genera calls for further revision of xyleborine classification.
Genomic organization and regulation of three Cecropin genes in Anopheles gambiae
Authors: Zheng X.‐L. , Zheng And L.
Published: March 2003

Three cecropin genes (AgCecA‐C) were identified from Anopheles gambiae, a major vector for malaria in sub‐Saharan Africa. These genes form a cluster with AgCecA and AgCecB positioned in opposite orientation, while AgCecC is downstream of AgCecA in the same direction. One intron is present in each of these three genes. Motif searches of promoter regions revealed elements that could be regulated by the NF‐κB family of transcriptional regulators. The divergent promoter (1186 nucleotides in length) between CecA and CecB and the promoter for CecC were analysed by transfection in An. gambiae cell lines. Results showed that these promoters were up‐regulated by lipopolysaccharide. The activity was further elevated when heat‐inactivated microbes were used to challenge the cell line. At least one NF‐κB site was required for inducible expression of both CecA and CecB.
The ‘balanced force’ concept for instrumentation of curved canals revisited
Authors: Charles  , Charles
Published: March 2003

The principal characteristics of K‐type files are described and the succession of operations as prescribed by the ‘balanced force’ method of preparation of root canals, particularly curved root canals, examined. On the basis of a simple mathematical model, the motions of a file in a straight canal and in a curved canal are successively considered, and the merits of the recommended method highlighted and confirmed.
Shaping ability of Profile rotary nickel‐titanium instruments with ISO sized tips in simulated root canals: Part 1
Authors: Bryant  , Thompson  , Al‐Omari  , Dummer
Published: March 2003

The aim of this study was to determine the shaping ability of ProFile 0.04 taper rotary nickel‐titanium instruments with ISO sized tips in simulated canals. A total of 40 simulated root canals made up of four different shapes in terms of angle and position of curvature were prepared by ProFile instruments using the ‘crown down’ approach recommended by the manufacturer. Part 1 of this two‐part report describes the efficacy of the instruments in terms of preparation time, instrument failure, canal blockages, loss of canal length and three‐dimensional canal form. The time necessary for canal preparation was on average 5.2 min and was not influenced significantly by canal shape. Three instrument fractures occurred and a further 3 instruments deformed; size 35 instruments failed the most (4) followed by size 30 (2), four of the failures occurred in canals with 40° curves. None of the canals became blocked with debris and change in working distance was minimal. Intra‐canal impressions of canal form demonstrated that most canals had apical stops and smooth canal walls whereas all canals had good flow and taper. Under the conditions of this study, ProFile 0.04 taper rotary nickel‐titanium instruments with ISO sized tips prepared simulated canals rapidly and created good three‐dimensional form.
Step‐by‐step description of a rotary root canal preparation technique
Authors: Schrader C. , Ackermann M. , Barbakow F.
Published: March 2003

Clinical technique Since the introduction of nickel‐titanium in endodontics, several canal preparation techniques involving the use of rotary instruments have become popular. Such engine‐driven rotary instruments rotate between 150 and 2000 r.p.m. and may be high or low torque orientated. This paper describes one such engine‐driven system called the ProFile technique. The instruments are of a different specification to that used for conventional endodontic files and reamers. This paper describes a technique employed by the Division of Endodontology,

Zurich Dental School, in a step‐by‐step procedure using primarily photographs of radiographs. The intention is to give any interested clinician a better idea of the technique using radiographs taken from both the buccolingual (clinical) perspective and the mesiodistal projection. Basically, the technique involves preparing the coronal portion of the root canal using Gates‐Glidden burs and the ProFile instruments. Only when any constricting coronal parts of the canals have been removed is the working length established using conventional files. Finally, the apical part of the canal is prepared using only the ProFile instruments. Three clinical cases are also briefly described, in order to illustrate the potential of the technique in cases treated generally by clinicians.
Prognosis in periradicular surgery: a clinical prospective study
Authors: Zuolo M. L. , Ferreira M. O. F. , Gutmann J. L.
Published: March 2003

Zuolo ML, Ferreira MOF, Gutmann JL. Prognosis in periradicular surgery: a clinical prospective study. International Endodontic Journal, 33, 91–98, 2000.

Aim  The purpose of this prospective study was to evaluate the prognosis of periradicular surgery using well‐defined case selection and a rigorous surgical protocol.

Methodology Teeth to be treated surgically demonstrated a periradicular lesion of strictly endodontic origin with or without clinical signs and symptoms of inflammation. A total of 114 teeth were treated. Following the reflection of a full mucoperiosteal tissue flap, residual soft tissues were curetted, root ends were resected with a fine high‐speed diamond bur, root‐end cavities were prepared ultrasonically with diamond tips, and IRM root‐end fillings were placed. Cases were

followed clinically and radiographically for a period ranging from 1 to 4 years.

Results  The results of this study showed 91.2% success out of a total of 102 teeth available for follow‐up, based on accepted parameters of evaluation. Cases were considered successful if there were no clinical signs or symptoms present and there was radiographic evidence of complete or incomplete healing (scar tissue). Factors related to case selection, parameters of healing and surgical technique are discussed in relation to the success rate identified in this prospective study.

Conclusions  Adherence to a strict endodontic surgical protocol and the use of contemporary techniques and materials will result in a predictably successful outcome in a wide range of teeth.
A laboratory study evaluating the release of hydroxyl ions from various calcium hydroxide products in narrow root canal‐like tubes
Authors: Larsen M. J. , Hörsted‐Bindslev P.
Published: March 2003

Aim

The aim of the present study was to describe pH changes in a variety of buffering solutions within a narrow test tube containing either a gutta‐percha point with incorporated calcium hydroxide, a commercial calcium hydroxide paste (Calcicur®) or a freshly mixed paste of calcium hydroxide in distilled water.

Methodology

The test material was placed centrally in a test tube of 2 mm inner diameter. Saline (1%) was placed at one end, whilst the buffering solutions were introduced at the other. The pH of the buffering solutions was monitored using electrodes placed at each end of the test tube.

Results

It was found that the pH 4.01 buffer strongly resisted pH changes at levels below 6.0, whilst saliva and bovine serum was buffered less and more evenly in the whole range up to pH 11.5. The calcium hydroxide containing gutta‐percha points caused the pH to increase quickly in the sodium chloride solution to levels above 11.5. However, in bovine serum, in saliva and in the pH 4.01 buffer the pH remained below 8.5, 8.0 and 6.0, respectively, 1 mm from the point. In contrast, the release of hydroxyl ions from the two calcium hydroxide pastes brought pH above pH 11.5 irrespective of the buffering of the solutions 5 mm from the paste.

Conclusion

It is concluded that Calcicur® and the calcium hydroxide‐water mixture contained substantially more available calcium hydroxide than did the calcium hydroxide containing gutta‐percha points, with the result that the release of hydroxyl ions from the points was limited in comparison to that from the pastes.
TGF‐β1 alone and in combination with calcium hydroxide is synergistic to TGF‐β1 production by osteoblasts in vitro
Authors: Jaunberzins A. , Gutmann J. L. , Witherspoon D. E. , Harper R. P.
Published: March 2003

Aim

To examine the effects of calcium hydroxide (Ca(OH)2), transforming growth factor‐beta (TGF‐β1), and Ca(OH)2/TGF‐β1 coadministration on TGF‐β1 and interleukin‐6 (IL‐6) synthesis by early (subculture 1) and late (subculture 5) osteoblast cultures.

Methodology

Early and late cultures were established using bone cells harvested from 21‐day‐old fetal rat calvaria. Cell cultures of both early and late osteoblasts were divided into four groups: group 1, control; group 2, cells challenged with Ca(OH)2; group 3, cells challenged with TGF‐β1; and group 4, cells challenged with Ca(OH)2 and TGF‐β1 in combination. TGF‐β1 and IL‐6 levels for all groups were determined using ELISA methodology.

Results

anova and Tukey HS analyses revealed that osteoblasts of groups 3 and 4 significantly increased (P < 0.001) TGF‐β1 synthesis in both early and late cultures of osteoblasts. IL‐6 was not detected in any of the groups considered in this study.

Conclusions

Exogenous TGF‐β1 has an autocrine effect on cell cultures of osteoblasts. Administration of TGF‐β1 alone or in combination with Ca(OH)2 increases the synthesis of TGF‐β1 in osteoblast cultures. Ca(OH)2 and TGF‐β1 are compatible when placed in a culture of osteoblasts. Ca(OH)2 provides a favourable environment for the anabolic effects of TGF‐β1.
Root canal treatment in mandibular canines with two roots: a report of two cases
Authors: D'Arcangelo C. , Varvara G. , De Fazio P.
Published: March 2003

Aim To highlight anatomical variation in the mandibular canine.

Summary Two clinical case reports are presented to illustrate anatomical variation in the human mandibular canine. Endodontic treatment may sometimes fail because morphological features of the tooth adversely affect the treatment procedures. Many investigators have reported the anatomical variations associated with mandibular canines. Mandibular canines are recognized as usually having one root and one root canal in most cases, although approximately 15% may have two canals or sometimes two roots. This paper describes two clinical cases of mandibular canines with two roots and two canals.

Key learning points

 Failure to control infection in the root canal system has an adverse impact on outcome.

 Clinicians should be aware of anatomical variations in the teeth they are managing, and should never assume that canal systems are simple.

 The majority of mandibular canines have one root and root canal, but 15% may have two canals, and a smaller number may have two distinct roots.
Clinical and radiographic evaluation of pulpotomies performed under intrapulpal injection of anaesthetic solution
Authors: Teixeira L. S. , Demarco F. F. , Coppola M. C. , Bonow M. L. M.
Published: March 2003

Aim

The purpose of this study was to evaluate clinically and radiographically pulpotomies carried out under intrapulpal injection of anaesthetic solution.

Methodology

Forty‐one permanent mandibular molar teeth presenting with deep carious lesions and/or exposed pulps, with or without periapical changes on radiographic examination, were treated with pulpotomy and dressed with calcium hydroxide. The teeth were divided into three groups. Group A consisted of 15 teeth, where intrapulpal anaesthesia was administered by a slow injection of lidocaine hydrochloride 2%. Group B, with 14 teeth, where intrapulpal anaesthesia was obtained with lidocaine hydrochloride 2% with adrenaline 1 : 100 000. Group C consisted of 12 teeth in which anaesthesia was performed with a mandibular block using prilocaine hydrochloride 3% with felypressin 1 : 100 000. Healing was evaluated using clinical and radiographic criteria: dentine barrier formation, absence of clinical symptoms and resolution of periapical involvement.

Results

After an observation time of 6–8 weeks (postoperative control) and 24–32 weeks (intermediate control), healing occurred in 13 teeth from group A (87%), in 11 teeth from group B (79%) and in 10 teeth from group C (83%). No statistical difference was demonstrated between the three groups (Fisher’s exact test).

Conclusions

Based on the methodology adopted, intrapulpal injection of anaesthetic solution did not impair healing in pulpotomized teeth.
Binding of seminal vesicle proteins to the plasma membrane of rat spermatozoa in vivo and in vitro
Authors: CARBALLADA  , ESPONDA
Published: March 2003

The association of seminal vesicle (SV) proteins with rat spermatozoa has been studied in vivo and in vitro. SV proteins bind to the sperm plasma membrane after ejaculation but are removed progressively from the sperm plasma membrane in the female genital tract. Although some of these remain bound to spermatozoa when they reach the oviducts, they do not seem to be present at the time of fertilization. This could indicate a putative role for these SV proteins in pre‐fertilization events. In addition, the binding of SV antigens was studied in vitro. It was observed that the ability to bind SV proteins is gained by the spermatozoa during epididymal maturation, and is first detectable in spermatozoa collected from the cauda epididymis. On the other hand, the binding is regulated by other proteins present in the ejaculate which are secreted by the coagulating glands. Experiments also showed that mouse spermatozoa are able to bind rat SV proteins, indicating that the binding is not a highly species‐specific phenomenon.
Effects of ethanol ingestion on epididymal glycosidases and fertility in the rat
Authors: SRIKANTH  , MALINI  , GOVINDARAJULU  , BALASUBRAMANIAN
Published: March 2003

Epididymal glycosidases play a role in sperm maturation by modifying sperm surface glycoproteins. To study the effects of ethanol on epididymal sperm maturation, ethanol (3 g/kg body weight as 25%, v/v) was administered to a group of rats by gastric‐intubation twice daily for 30 days. In another group, rats were also treated with alcohol for 30 days but were then withdrawn from treatment for 30 days to assess the reversibility of ethanol‐induced effects. Ethanol‐induced changes in epididymal tissue and sperm glycosidases, cauda epididymal sperm motility and the fertility of rats were assessed. Ethanol treatment caused a marked decrease in the specific activities of glycosidases in both tissues and spermatozoa from epididymal segments. Cauda epididymal sperm motility and the fertility of ethanol‐treated rats were significantly impaired compared to control rats fed an isocaloric diet. These changes are likely to be the consequence of direct and indirect effects of ethanol mediated through subnormal testosterone and dihydrotestosterone. Most of these changes were found to be reversible. The present study suggests that impaired activity of sperm glycosidases may be one of the factors responsible for defective sperm motility and fertilizing potential in ethanol‐treated rats.
The maturation of sperm motility in the epididymis and vas deferens of the vervet monkey,Cercopithecus aethiops
Authors: HORST GERHARD VAN DER , SEIER JURGEN V. , SPINKS ANDREW C. , HENDRICKS SHAMILA
Published: March 2003

Among the diverse facets of sperm maturation, changes in motility are conspicuous and hence studies of sperm kinematics might provide good indices for sperm maturation. Accordingly, the maturation of sperm motility in the epididymis and vas deferens of the vervet monkey,Cercopithecus aethiops, was assessed using a computer‐aided sperm motility analysis system. The results revealed clear trends in the development of both sperm motilityper seand in the movement characteristics of motile spermatozoa from different regions of the epididymis, the vas deferens and the ejaculate, reflecting maturational changes associated with the attainment of functional motility and fertility. Motion of spermatozoa from the caput epididymis was sluggish and irregular. As the spermatozoa moved through the corpus epididymis, motility increased sharply, and continued to improve through the cauda epididymis and vas deferens. Despite the high proportion of motile cells, full maturation of motion capabilities was not completed in spermatozoa from the corpus epididymis. Only once spermatozoa had reached the cauda epididymis and vas deferens did they attain their full vigour, and swam rapidly (greater VCL, VSL and VAP) with straightline trajectories (greater LIN, WOB and STR; lower ALH, MAD and CURV). After acquiring their maximal percentage motility and progressive velocity in the cauda epididymis and vas deferens, a slight decline in motility and vigour occurred in ejaculated spermatozoa, and was possibly associated with the ageing of stored spermatozoa. The results from this investigation have revealed clear trends in the maturation of the motility of vervet monkey spermatozoa during their transit through the epididymis and vas deferens and final emergence in the ejaculate, and have provided crucial baseline information on the reproductive physiology of this potentially valuable biomedical model to serve as a reference for future studies in reproductive toxicology.
Spermatogonial transplantation technique in spermatogenesis research
Authors: Ogawa Takehiko
Published: March 2003

The spermatogonial stem cell is the foundation of spermatogenesis and it continues to divide throughout the lifetime of a male. It divides to renew itself as well as to produce daughter cells that finally differentiate to spermatozoa. Transplantation of spermatogonial stem cells from a fertile mouse to the testis of a sterile mouse results in the development of donor cell derived spermatogenesis. Furthermore, spermatogenesis of the rat and hamster occurs in immunodeficient mouse testes following spermatogonial transplantation. Cryopreservation of spermatogonial stem cells has been shown to be applicable in many species for later transplantation. In addition, certain forms of infertile donor germ cell can be transplanted to produce functional sperm in a recipient testis. These new techniques will be useful for basic research of spermatogenesis and for application to reproductive technology in the future.
Enrichment and transplantation of spermatogonial stem cells
Authors: Shinohara Takashi , Brinster Ralph L.
Published: March 2003

Spermatogenesis is a complex, highly organized process originated from stem cell spermatogonia. Because there are very few stem cells and they can only be defined by their function, the identification and isolation of these cells has been very difficult. By using a spermatogonial transplantation assay system, we have identified α6‐and β1‐integrin expression on stem cells, and cells isolated with these antigens were significantly enriched in stem cells. This is the first demonstration of spermatogonial stem cell‐associated antigens. Analysis of two infertile mouse models, Steel/SteelDickie (Sl/Sld) and experimental cryptorchidism, showed that the number of stem cells is reduced in Sl/Sld testis. Whereas cryptorchid testes are greatly enriched for stem cells, and one in 200 cells is a stem cell. These techniques will provide an important starting point for further purification and characterization of spermatogonial stem cells.
Computer‐assisted assessment of sperm morphology: comparison with conventional techniques
Authors: Marnet B. , Vieitez G. , Milhet P. , Richoilley G. , Lesourd F. , Parinaud J.
Published: March 2003

The aim of the present study was to compare conventional and computer‐assisted morphology assessment of spermatozoa. Sixty‐two semen samples from patients undergoing in vitro fertilization (IVF) and 40 samples from patients undergoing an intracytoplasmic sperm injection (ICSI) were studied using both techniques. The percentage of normal spermatozoa found was closely correlated between the techniques (r=0.788, p < 0.0001). The intra‐operator variation was low for both techniques but the inter‐operator variation was much higher with the conventional than with the computer‐assisted method (coefficient of variation = 0.43 vs. 0.08, respectively, for conventional and computer‐assisted assessments). The percentage of spermatozoa with normal morphology, as well as sperm motility, was significantly enhanced after PureSperm preparation, whatever the method used for assessment. In the IVF study, fertilization rate was poorly correlated with sperm morphology using both methods. However, combined with motility, morphology assessed with the computer allowed discrimination of two groups of patients with significantly different fertilization rates (30.5 ± 5.4% vs. 63.1 ± 5.4%, p < 0.0001). In contrast, the fertilization rate in ICSI was influenced neither by sperm morphology nor by motility. In conclusion, computer‐assisted assessment of sperm morphology has a slightly better predictive value for ART than conventional assessment, but above all is much more reproducible, allowing standardization.
pH and bicarbonate in the ductuli efferentes testis of the rat
Authors: Newcombe N. , Clulow J. , Man S. Y. , Jones R. C.
Published: March 2003

The pH and bicarbonate concentrations of luminal fluids in the efferent ducts of the rat were estimated from pH measurements of samples in vitro under conditions of controlled temperature and carbon dioxide tension. The pH of scrotal blood was estimated to be more acidic than systemic blood (mean pH=7.44) at either of the putative scrotal carbon dioxide tensions (5% and 7%, pH, respectively,=7.42 and 7.28). For PCO2 tensions of 5% and 7%, respectively, the data indicated that the pH in the efferent ducts was significantly higher (distal initial zone pH=7.55 or 7.41; coni vasculosi pH=7.66 or 7.51; p < 0.01) than in fluid entering (rete testis fluid, pH=7.34 or 7.20) or leaving the ducts (zone 1a of the epididymal duct 7.26 or 7.11). Bicarbonate concentrations were also significantly higher (p < 0.01) in the efferent ducts (35.4 ± 4.7 mM, distal initial zone; 45.2 ± 7.6 mM, coni vasculosi) than in fluids entering (22.9 ± 3.6 mM) or leaving (20.4 ± 4.9 mM) the ducts. Estimates of the reabsorption of bicarbonate and fluid indicated that 96% of the testicular output of bicarbonate was reabsorbed in the efferent ducts, but there was also some secretion of bicarbonate into the ducts. It is concluded that luminal pH and bicarbonate levels in the efferent ducts of the rat are high relative to those found in the epididymis where low pH and bicarbonate contributes to sperm quiescence during storage. Nevertheless, the high rate of bicarbonate reabsorption in the efferent ducts is a major contributor to the establishment of the low pH and bicarbonate milieu of the epididymis.
Morphometric evaluation of the human prostate
Authors: Arenas María I , Romo Eva , Royuela Mar , Ruiz Antonio , Fraile Benito , Sánchez‐Chapado Manuel , Paniagua Ricardo
Published: March 2003

In order to clarify the ageing‐related histological changes in the human prostate, a quantitative morphometric analysis was performed. Complete prostates were obtained at autopsy from 281 men (aged 20–84 years) who died in traffic accidents and presented no clinical symptoms of prostatic disease. The prostates were classified as: histologically normal (n=182), with nodular hyperplasia (n=42), with intraepithelial neoplasia (n=40) and carcinomatous with low Gleason grade (n=20). Each prostate was divided into three regions (periurethral, central and peripheral) and the volume of each region, as well as the average volume occupied by stroma and epithelium in each region were quantified. For each parameter, the average values for each age group were compared. In the histologically normal prostates, an increase with ageing in the total volume and the volume occupied by the central region were observed; these increases were mainly caused by an increase in the stromal volume of the central region in men after 30 years of age. No histologically normal prostates were found in men older than 70 years of age. Nodular prostatic hyperplasia was found in men over 30 years of age and a fluctuation in the total volume throughout ageing was observed. Prostates with intraepithelial neoplasia (PIN) and carcinoma were observed in men aged >20 years and the total volume and those of each prostatic region showed multiple variations, except for the eighth decade where a marked increase with regard to that of the previous decades was observed.