bluuebunny/crossref_metadata_2025 huggingface dataset:
A senior executive end‐user framework
Authors: Seeley Monica E. , Targett David
Published: November 2003

Senior executives make decisions about their organization's information technology (IT) budgets that often involve substantial expenditure. Yet, in terms of their own competence in using IT, there is evidence to suggest that they may not be the best members of the organization to make such decisions. This paper reports on a study of 85 senior executives across 16 large organizations and the depth and breath of applications these executives use to support them at an individual level. The research design and methodologies were based on the principles of grounded theory. The primary source of data collection was in‐depth interviews. A number of end‐user frameworks have been developed by previous authors; however, most are more pertinent to middle and junior managers. Based on the analysis of the data from this study an end‐user framework is proposed that is more appropriate for senior executives.
The reality of participative systems design: contributing to stability in a rocking boat
Authors: Mumford Enid
Published: November 2003

This paper suggests that participative design will only be accepted by industry if it can be shown to contribute both to greater efficiency and to more satisfied and productive employees. It compares the introduction of socio‐technical systems into US, Japanese, Scandinavian and British industry and suggests that this approach was used with most enthusiasm in those countries that had a shortage of labour.
Mitochondrial DNA sequence divergence in weevils of the Pissodes strobi species complex (Coleoptera: Curculionidae)
Authors: Langor D. W. , Sperling F. A. H.
Published: November 2003

Mitochondrial DNA sequence divergence is unusually high between several species of the Pissodes strobi complex, in contrast to low allozyme and morphological divergences, and the ability to hybridize in the laboratory. We sequenced an 810 bp segment in seven individuals, representing four species in the P. strobi complex and one outgroup species, P. affinis Randall. The 810 bp segment covered the 3′ half of the cytochrome oxidase subunit I (COI) gene. We also sequenced one specimen of P. strobi (Peck) over a 2301 bp region of mtDNA, extending from the 5′ end of COI to the 3′ end of COII. Uncorrected sequence divergences were below 1.1% among three specimens of P. schwarzi Hopkins, and between P. terminalis Hopping and P. nemorensis Germar. All other interspecific combinations in the P. strobi complex showed divergences of 6.0–7.5%. The outgroup species, P. affinis, had an average divergence of 12.8% from members of the P. strobi group. As in most other insects, A+T content in Pissodes mtDNA was high; transition:transversion ratio was high among lineages exhibiting low divergences, but declined with increasing sequence divergence; and inferred amino acid sequence divergences were low. The degree of sequence divergence differs markedly across different functional regions of the COI gene. The high mtDNA divergences within the P. strobi complex calculated from direct sequencing support earlier reports based on restriction site surveys; however, cladograms based on mtDNA sequence differ from those based on earlier work.
Phylogenetic relationships of the Alysiinae/Opiinae (Hymenoptera: Braconidae) and the utility of cytochrome b, 16S and 28S D2 rRNA
Authors: Gimeno C. , Belshaw R. , Quicke D. L. J.
Published: November 2003

Subfamily to generic level relationships among the Opiinae and Alysiinae (Hymenoptera: Braconidae) were investigated using DNA sequence data from c. 400 bp fragments of the cytochrome b, 16S rRNA and 28S D2 rRNA genes. The best recovery of morphologically‐defined groups was achieved by 28S D2, followed by 16S with cytochrome b being very uninformative. We suggest ribosomal expansion regions may prove useful for this level of phylogenetic reconstruction. Combined analyses on all genes (using several alignment and phylogenetic methods) improved resolution but failed to resolve the proposed paraphyly of the Opiinae and Alysiini. Support was found for other relationships within the Opiinae and, contrary to a previous morphological study, there was strong evidence against the Gnamptodontinae being the sister group of the Opiinae/Alysiinae clade.
The white gene from the yellow fever mosquito, Aedes aegypti
Authors: Coates C. J. , Schaub T. L. , Besansky N. J. , Collins F. H. , James A. A.
Published: November 2003

We report the cloning and primary characterization of both cDNA and genomic fragments from the white gene of the yellow fever mosquito, Aedes aegypti. Comparisons of the conceptual translation product with white genes from four other species within the order Diptera show that the Ae. aegypti gene is most similar to the white gene of the mosquito vector of human malaria, Anopheles gambiae (86% identity and 92% similarity). The analysis of the primary sequence of genomic DNA at the 5′‐end of the coding region revealed the presence of an intron that is also present in An. gambiae, but not in the vinegar fly, Drosophila melanogaster. The isolated clones of the Ae. aegypti white gene will enable the construction of a marker gene for use in the development of a germline transformation system for this species.
Cloning and characterization of three Musca domestica yolk protein genes
Authors: White N. M. , Bownes M.
Published: November 2003

The yolk protein (yp) genes encode the major nutritional polypeptides deposited in developing oocytes for subsequent utilization during embryogenesis, and represent a highly conserved family of genes in higher Diptera. Originally isolated from Drosophila melanogaster, they are expressed in a temporal‐, tissue‐ and sex‐specific manner in all species in which they have been identified. We report here the isolation of cDNAs encoding three independent yolk proteins from the common housefly, Musca domestica. Expression of the three M. domestica yp genes is analysed both by Northern and in‐situ hybridization. We discuss in an evolutionary context both the significance of the expression patterns, and regions of apparent polypeptide sequence divergence.
Double‐strand conformation polymorphism (DSCP) analysis of the mitochondrial control region generates highly variable markers for population studies in a social insect
Authors: Atkinson L. , Adams E. S.
Published: November 2003

Genetic markers were obtained for the termite Nasutitermes corniger by DSCP (double‐strand conformation polymorphism) analysis of PCR‐amplified mitochondrial control region DNA. This procedure revealed twenty‐one haplotypes in forty‐four colonies, whereas a restriction fragment length polymorphism analysis detected only nine haplotypes. Sequence analysis of DSCP fragments of contrasting mobilities suggests that the electrophoretic haplotypes are caused by DNA curvature in this highly AT‐rich region. DSCP markers showed that some termite colonies contained maternally unrelated queens, each of which produced worker offspring. This pattern is consistent with nest founding by unrelated queens. Due to the availability of conserved primers for the mtDNA control region, DSCP analysis may readily reveal comparatively high levels of variation in a wide variety of organisms.
Treatment of root perforations with calcium hydroxide and SuperEBA cement: a clinical report
Authors: BOGAERTS P.
Published: November 2003

Iatrogenic root perforations sometimes occur during root canal treatment or when preparing the root canal for a post. The inflammation in the periradicular tissues caused by root perforation reduces the prognosis of the involved tooth. The situation may be further complicated if by coronal leakage, microorganisms are allowed to infect the canal and the periradicular region. This clinical report presents a nonsurgical technique, based on the internal matrix concept, using both calcium hydroxide and SuperEBA together to repair root perforations. The author has experienced good clinical results with this perforation repair technique.
Comparison of two laser Doppler systems on the measurement of blood flow of premolar teeth under different pulpal conditions
Authors: MESAROS S. , TROPE M. , MAIXNER W. , BURKES E. J.
Published: November 2003

An in vivo investigation was conducted to assess the ability of two laser Doppler flowmeters (DRT4, Moor Instruments Ltd; Laserflo BPM2, Vasamedics) to differentiate between vital pulp, ischaemically necrosed pulp, and an empty pulp chamber in premolar teeth in children. Sixteen intact, vital teeth in need of extraction for orthodontic purposes were identified. Laser Doppler blood flow readings were taken prior to (Tl) and after the administration of adrenaline‐free local anaesthetic (T2), after extraction and replantation of the teeth (T3), and after broaching the pulp tissue and replanting the teeth (T4). The data collected (flux values) far both instruments were analysed using a repeated measures ANOVA. Significant differences were found for vital pulp (Tl, T2) versus ischaemically necrosed pulp (T3), and for vital pulp (Tl, T2) versus an empty pulp chamber (T4), with the Moor DRT4 instrument This instrument also detected a significant difference in pulpal blood flow to the vital pulp, before and after the administration of local anaesthetic. No other significant differences were found.
Periapical radiographic techniques during endodontic diagnosis and treatment
Authors: FAVA L. R. G. , DUMMER P. M. H.
Published: November 2003

The objective of this article is to describe various radiographic projections which can be used during endodontic therapy. Changes to the angulation of the X‐ray beam in relation to the teeth and film can help diagnosis and treatment by producing images which provide additional information not always visible on radiographs taken with standard angulations. For example, changes in angulation can be useful to determine the number and curvature of roots and canals, to identify superimposed roots and to distinguish between anatomical landmarks and apical pathology. Although use of such techniques increases the diagnostic yield of films, it must be appreciated that such views lead to images that are less distinct because of inherent image distortion. Nevertheless, use of the various techniques during endodontics can provide substantial benefit for clinicians In their daily practice.
Effect of sealer thickness on long–term sealing ability: a 2–year follow–up study
Authors: KONTAKIOTIS E.G. , WU M‐K. , WESSELINK P.R.
Published: November 2003

Long‐term sealing ability of root‐canal sealer is of clinical importance. We compared the performance of five sealers and a dentine bonding agent, at thicknesses of 0.05 mm (thin layer) and 0.3 mm (thick layer), in 212 standard human root specimens, before and after storage in water for 2 years. Thick layers of Roth and Pulp Canal Sealer EWT (PCS–EWT) allowed more leakage than did thin layers (P = 0.0000, 0.0358 for Roth and PCS–EWT, respectively) whereas no significant difference was found between the thin and thick layers for AH26, Ketac‐Endo, Sealapex and J& J Bonding Agent. The pooled results of the thin and thick layers, which have clinical relevance, showed that after storing the specimens in water for 2 years, AH26, Sealapex, Ketac‐Endo and PCS–EWT leaked more than before storage in water (P = 0.0008, 0.0000, 0.0035, 0.0257 for AH26, Sealapex, Ketac‐Endo and PCS–EWT respectively), and that)&J Bonding Agent allowed less leakage and PCS‐EWT more leakage than the other four sealers (P= 0.0000).
Scanning electron microscope study comparing four root canal preparation techniques in small curved canals
Authors: HEARD F. , WALTON R.E.
Published: November 2003

Various instrumentation techniques have been proposed and examined with conflicting results. They include hand and ultrasonic techniques and combinations of the two. In the present study we assesed the effectiveness of four preparation methods for cleaning small, curved root canals, using backscattered‐imaging scanning electron microscopy (SEM). The methods were: (i) step‐back without initial coronal flaring; (ii) step‐back with coronal flaring; (iii) step‐back with initial coronal flaring and finished by ultrasonic irrigation; and (iv) ultrasonics only.

Eighty freshly extracted maxillary and mandibular molars were randomly placed into four treatment groups of 20 teeth each. After preparation, roots were sectioned longitudinally and examined wet by SEM. Each canal was qualitatively evaluated and the groups compared for removal of debris and smear layer, both overall and at each level (apical, middle and coronal).

There were no statistically significant differences between the techniques, either overall or within any of the regions. When comparing regions (regardless of technique) the middle level was cleaner than the apical or coronal levels.

In conclusion, efficacy differed little among the techniques; none of them completely removed smear layer and all left debris.
Influence of infection at the time of root filling on the outcome of endodontic treatment of teeth with apical periodontitis
Authors: SJÖGREN U. , FIGDOR D. , PERSSON S. , SUNDQVIST G.
Published: November 2003

This study investigated the role of infection on the prognosis of endodontic therapy by following‐up teeth that had had their canals cleaned and obturated during a single appointment. The root canals of 55 single‐rooted teeth with apical periodontitis were thoroughly instrumented and irrigated with sodium hypochlorite solution. Using advanced anaerobic bacteriological techniques, post–instrumentation samples were taken and the teeth were then root‐filled during the same appointment. All teeth were initially infected; after instrumentation low numbers of bacteria were detected in 22 of 55 root canals. Periapical healing was followed up for 5 years. Complete periapical healing occurred in 94% of cases that yielded a negative culture. Where the samples were positive prior to root filling, the success rate of treatment was just 68%– a statistically significant difference. Further investigation of three failures revealed the presence of Actinomyces species in each case; no other specific bacteria were implicated in failure cases. These findings emphasize the importance of completely eliminating bacteria from the root canal system before obturation. This objective cannot be reliably achieved in a one‐visit treatment because it is not possible to eradicate all infection from the root canal without the support of an inter–appointment antimicrobial dressing.
Observation of Saccharomyces cerevisiae in blood of patient undergoing root canal treatment
Authors: DEBELIAN G. J. , OLSEN I. , TRONSTAD L.
Published: November 2003

In this study, an unusual observation of Saccharomyces cerevisiae isolated from an infected root canal and from the blood of a patient undergoing endodontic therapy of a tooth with asymptomatic apical periodontitis is reported. Phenotypic (biochemical tests, antifungal susceptibility tests and SDS‐PAGE of cellular proteins) and genetic (ribotyping) methods were used to characterize the strains. By using these methods it was found that the blood and root canal isolates were identical but differed from S. cerevisiae strains of other sources. It was therefore more than likely that the root canal was the source of the blood isolate and that it had been transferred unintentionally to the bloodstream during root canal treatment.
Analysis of forces developed during root canal preparation with the balanced force technique
Authors: BLUM J.‐ Y. , MACHTOU P. , ESBER S. , MICALLEF J. P.
Published: November 2003

The aim of this study was to examine the forces and torque developed during root canal preparation with the balanced force technique using a recently described force‐analyser device. A tooth was placed in a holder within the Endograph and forces and torques exerted were recorded. These parameters, which can be studied during preparation (on‐line) or stored and examinated subsequently (off‐line) generated endograms, which showed the forces generated with time. In addition, the endograms of preparations performed by students and endodontists, as well as deliberately induced failures in preparation technique (broken instruments), were compared. The values for the forces and torques depended on the size of the instruments and were related to the phase of the preparation. For the endodontists, the vertical and horizontal forces varied, respectively, from 0.08 ±0.01 kg for a size 15 to 0.65 ±0.10 kg for a size 45, and from 0.01 ±0.005 kg for a size 15 to 0.4±0.1 kg for a size 40. The torque varied from 0.08 ± 1 kg mm‐1 for a size 15 to 1.6 ± 0.4 kg mm‐1 for a size 45. With the Endograms used as a reference, the relation between the developed vertical forces and the torque became more similar between the groups of endodontists and students. The Endograph provides a new approach to the analysis of preparation technique because it depicts the relationships between the different parameters of the preparation.
Identification and characterization of antioxidant enzyme mRNAs in the rat epididymis
Authors: ZINI A. , SCHLEGEL P. N.
Published: November 2003

Spermatozoa are highly sensitive to oxidative stress. The epididymis, a natural sperm reservoir, has maturational and storage functions. The epididymis may also protect spermatozoa from oxidative injury by elaborating scavengers of reactive oxygen species (ROS). Therefore, we have evaluated the mRNA expression of antioxidant enzymes in the normal rat epididymis and the effects of efferent duct ligation on the expression of these enzymes.

Adult rat epididymides were harvested, divided into caput, corpus and cauda and processed for RNA extraction. Additional adult rats were subjected to unilateral efferent duct ligation and the epididymides harvested at 1, 4, 8, 16 or 28 days after the procedure. Antioxidant enzyme mRNA expression was assessed by Northern blot analysis using 32P‐labelled DNA probes derived from known cDNA sequences for classical cellular glutathione peroxidase (GSHPx), phospholipid hydroperoxide glutathione peroxidase (PHGPX), secretory epididymal glutathione peroxidase (E‐GPX), copper‐zinc superoxide dismutase (SOD), secretory epididymal superoxide dismutase (E‐SOD) and catalase. Specific mRNA levels were measured, with gene expression evaluated relative to total RNA, not per organ. Variations in lane loading were controlled by measuring the levels of 28S ribosomal RNA.

GSHPx, PHGPX, SOD and catalase mRNA were detected in the caput, corpus and cauda epididymis. E‐GPX mRNA was only detected in the caput, whereas E‐SOD mRNA was primarily detected in the corpus. At 28 days after efferent duct ligation, epididymal weight decreased by 34% relative to controls ( p <  0.05). With the exception of PHGPX, the relative mRNA levels of the antioxidant enzymes studied did not change after efferent duct ligation.

This study demonstrates that mRNAs for multiple antioxidant enzymes are expressed in the epididymis and that the relative expression of these enzymes remains largely unchanged in response to efferent duct ligation. Taken together, these results suggest that antioxidant enzymes may play an important, region‐specific role in epididymal function. Expression of the secretory antioxidant enzymes E‐SOD and E‐GPX is region‐specific, indicating that the need for antioxidant enzymes may vary along the length of the epididymis.
Evaluation of sperm fertilizing ability using the Sperm Quality Analyzer
Authors: SHIBAHARA H. , NAITO S. , HASEGAWA A. , MITSUO M. , SHIGETA M. , KOYAMA K.
Published: November 2003

The Sperm Quality Analyzer is an inexpensive device which provides a quantitative estimation of sperm motility. To evaluate the fertilizing ability of human spermatozoa using a Sperm Quality Analyzer, correlations amongst the sperm motility index, the sperm penetration index (as assessed using the sperm penetration assay; SPA), and the fertilization rate in the treatment of standard IVF‐ET were analysed retrospectively. The sperm motility index demonstrated a significant correlation with sperm concentration ( p < 0.001), sperm motility ( p < 0.001) and the motile sperm concentration ( p < 0.001) in a total of 104 fresh semen samples from 81 men donating samples for IVF‐ET. The sperm motility index also showed a significant correlation ( p < 0.001) with the sperm penetration index in 60 patients, assessed using the SPA, before they were treated by standard IVF‐ET. The correlation between the sperm motility index and the IVF‐ET fertilization rate was higher than that between the sperm penetration index and the fertilization rate. The sperm motility index was classified into three categories: `poor' (sperm motility index < 80), `medium' (sperm motility index 81–160) and `good' (sperm motility index>  160). The relationships between the IVF‐ET fertilization rate and each category of the sperm motility index values were also evaluated. For the three categories in the sperm motility index, the fertilization rates (76.0%) of 60 samples judged as `good' were significantly higher than those (44.2%) of 15 samples judged as `medium' ( p < 0.001) and those (34.7%) of 13 samples judged as `poor' ( p < 0.001). These results indicate that the Sperm Quality Analyzer provides a reliable estimation of the fertilizing ability of human spermatozoa.
Reactive oxygen species and human spermatozoa: physiology and pathology
Authors: GRIVEAU J. F. , LANNOU D. LE
Published: November 2003

The role of reactive oxygen species (ROS) in the pathophysiology of human sperm function has been emphasized in recent years. ROS production in semen has been associated with loss of sperm motility, decreased capacity for sperm–oocyte fusion and loss of fertility. There is a current presumption that the most prolific source of ROS in sperm suspensions is an NADPH oxidase located in leukocytes or in spermatozoa which produces superoxide which is further converted to peroxide by the action of superoxide dismutase. Hydrogen peroxide has been recognized as the most toxic oxidizing species for human spermatozoa, which are very sensitive to lipid peroxidation owing to the high content of polyunsaturated fatty acids in their plasma membrane, though this is not the sole mechanism by which sperm function might be impaired by ROS. Although the excessive production of ROS is detrimental to human spermatozoa, there is a growing body of evidence which suggests that ROS are also involved in the physiological control of some sperm functions. This review focuses on the nature and source of the ROS generated by human spermataozoa as well as their operational mechanisms and their effects, which may be detrimental or beneficial.
Pituitary, gonadal and adrenal hormones after prolonged residence at extreme altitude in man
Authors: BASU M. , PAL K. , PRASAD R. , MALHOTRA A. S. , RAO K. S. , SAWHNEY R. C.
Published: November 2003

High altitude‐induced alterations in pituitary, gonadal and adrenal hormones were studied in (i) eugonadal men from the armed forces who were resident at sea level (SL), (ii) SL residents staying at an altitude of 3542 m for periods ranging from 3 to 12 months (acclimatized lowlanders, ALL), (iii) ALL who stayed at 6300 m for 6 months, (iv) ALL who trekked from 3542 to 5080 m and stayed at an altitude of more than 6300 m in the glacier region for 6 months, and (v) high‐altitude natives (HAN) resident at an altitude of 3300–3700 m. Circulating levels of LH, FSH, prolactin, cortisol, testosterone, dihydrotestosterone (DHT) and progesterone in ALL at 3542 m and in HAN were not significantly different ( p > 0.05) from the SL control values. When the ALL living at 3542 m trekked to an extreme altitude of 5080 m, their testosterone levels showed a significant decrease ( p < 0.01) compared to the preceding altitude values but had returned to SL values when measured after 6 months' continuous stay at 6300 m. As with testosterone, the levels of DHT and oestradiol‐17β (E2) after prolonged stay at extreme altitude were also not significantly different ( p > 0.05) from the SL values. The LH levels after trekking to 5080 m were significantly higher ( p < 0.01) than at an altitude of 3542 m, but decreased to levels found at 3542 m or SL after prolonged residence at extreme altitude. Plasma levels of ACTH, prolactin, FSH and cortisol on arrival at 5080 m, and after a 6‐month stay at extreme altitude, were not significantly different ( p > 0.05) from the SL values. Plasma progesterone levels tended to increase on arrival at 5080 m but a significant increase ( p < 0.001) was evident only after a 6‐month stay at extreme altitude. These observations suggest that prolonged residence at lower as well as at extreme altitude does not appreciably alter blood levels of pituitary, gonadal or adrenal hormones except for plasma levels of progesterone. The exact mechanism and significance of this increase remains unknown, but may be important in increasing the sensitivity of the hypoxic ventilatory response and activation of haemoglobin synthesis.
Influence of oxygen tension on reactive oxygen species production and human sperm function
Authors: GRIVEAU J. F. , LANNOU D. LE
Published: November 2003

Human spermatozoa were exposed to the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) to stimulate endogenous production of reactive oxygen species (ROS). They were incubated under a gas phase of 5% CO2/90% N2/5% O2, or 5% CO2/95% air (20% O2) to investigate whether a lower than atmospheric oxygen tension in the gas phase of the incubator limited the endogenous production of ROS by human spermatozoa and thus was able to reduce the cytotoxic effects of ROS on sperm function. Exposure of human spermatozoa to exogenous NADPH induced an 8‐fold higher production of superoxide anion under ambient vs. low oxygen tension. This marked difference in the stimulation of superoxide anion generation was associated with significantly different sperm motility parameters, according to the oxygen tension in the gas phase of the incubator. Whereas under 5% oxygen the percentage of motile spermatozoa was unaffected by the presence of NADPH, all of the motility parameters recorded under an atmosphere of 5% CO2 in air were dramatically affected, not only compared to their respective controls, but also compared to the motility parameters observed under low oxygen tension. The presence of superoxide dismutase plus catalase protected spermatozoa against the toxic effects of NADPH, confirming a cause/effect relationship between the increased superoxide production and reduced sperm function. Even at a concentration of NADPH which did not alter the percentage of motile spermatozoa, hyperactivated motility and acrosome reaction were significantly lower under an atmosphere of 5% CO2 in air compared to a gas phase of 5% CO2/90% N2/5% O2. These results suggest that there is an advantage in using 5% O2 rather than 20% O2 in the gas phase of the incubator to prevent the excessive production of ROS by spermatozoa and related alterations of sperm functions. This may be of clinical value in fertilization programmes.
Regulation of γ‐glutamyl transpeptidase activity by Ca2+‐ and protein kinase C‐dependent pathways in Sertoli cells
Authors: MERONI S. , CÁNEPA D. , PELLIZZARI E. , SCHTEINGART H. , CIGORRAGA S.
Published: November 2003

γ‐Glutamyl transpeptidase (γ‐GTP) activity in Sertoli cells can be stimulated by FSH. This cAMP‐dependent metabolic event can be enhanced when Sertoli cells are co‐cultured with germ cells, suggesting that different signal transduction pathways may be involved in the regulation of γ‐GTP activity. In this study we examined the participation of Ca2+‐ and protein kinase C (pkC)‐dependent signal transduction pathways in the regulation of basal and FSH‐stimulated γ‐GTP activity. Under basal conditions, the increase in extracellular Ca2+ concentration or the addition of the Ca2+ ionophore 4Br‐A23187 produced a decrease in γ‐GTP activity. Conversely, blockage of voltage‐dependent Ca2+ channels with verapamil or nifedipine or inhibition of Ca2+‐calmodulin dependent processes with trifluoperazine resulted in an increase in γ‐GTP activity. To study the role of a pkC‐dependent pathway the effects of low doses of staurosporine were evaluated. Under these experimental conditions an increase in γ‐GTP activity was observed. It was then investigated whether these signal transduction pathways could interact with the FSH‐stimulated cAMP‐dependent pathway to regulate γ‐GTP activity. Increase in extracellular Ca2+ concentration, the addition of 4Br‐A23187 or the blockage of voltage‐dependent Ca2+ channels did not modify FSH‐stimulated γ‐GTP activity. However, staurosporine produced an additional increase in FSH‐stimulated γ‐GTP activity and this effect was also observed when cells were stimulated with dbcAMP. In summary, our data are consistent with an inhibitory role of Ca2+‐calmodulin‐ and pkC‐dependent pathways in the regulation of basal γ‐GTP activity. Similar to what has been shown for other Sertoli cell parameters, a pkC‐dependent pathway can interact with the FSH‐stimulated cAMP‐dependent pathway. The precise steps involved in this interaction are still unknown.
Evidence that peptides derived from the disintegrin domain of primate fertilin and containing the ECD motif block the binding of human spermatozoa to the zona‐free hamster oocyte
Authors: GICHUHI P. M. , FORD W. C. L. , HALL L.
Published: November 2003

The binding of human spermatozoa to, and penetration of, zona‐free hamster oocytes were significantly inhibited in the presence of (i) a 28‐residue peptide corresponding to part of the disintegrin‐like domain of monkey fertilin β, or (ii) a hexapeptide containing the ECD motif found in the disintegrin‐like domain of monkey and human fertilin β, or (iii) a hexapeptide containing an RGD motif, implicated in integrin recognition. Polyclonal antibodies raised against the 28‐residue peptide were used in conjunction with immunobeads to demonstrate that this peptide bound to the oolemma of zona‐free hamster oocytes. These data support the view that the ECD motif is involved in the recognition of the oolemma receptor by human fertilin.
Histochemical demonstration of zinc ions in ejaculated human semen
Authors: STOLTENBERG M. , SØRENSEN M. B. , DANSCHER G.
Published: November 2003

A revised in‐vitro technique for autometallographic demonstration of chelatable zinc in the human ejaculate is presented, and the localization of the loosely bound pool of zinc ions is described in semen smears and at the ultrastructural level. In semen smears, black autometallographic (AMG) grains indicated the presence of zinc ions dispersed between the spermatozoa. These AMG grains have the same size as grains associated with the sperm tail and may have the same origin. EM analysis of AMG‐developed smears fixed in osmium suggested that the detected zinc ions might be related to huge protein molecules present in semen and adhering to the surface of the spermatozoa.

Spermatozoa in AMG‐stained smears exhibited zinc ions in the midpiece and head, and also joined to the membrane of the tail. Washed spermatozoa exhibited zinc ions only within the midpiece. Ultrastructurally, they were found located in the helecine mitochondria. A few grains were found in the acrosome of the washed spermatozoa. Treatment with the chelating agent DEDTC resulted in complete bleaching of the zinc staining. These findings and the fact that calcium EDTA acid blocks the plasma and surface staining, but not the acrosomal and mitochondrial staining, suggest that chelatable zinc ions exist in two separate pools in human semen.
Enlarged spectrum of seminological diagnoses using the resazurin colour reaction; a spectrophotometric application
Authors: RAHMAN N. A. , KULA K.
Published: November 2003

Two sets of investigations were carried out in the present study to establish whether, in semen samples processed for the resazurin reaction (RT), the colour change and its intensity were informative for both the quality and quantity of spermatozoa. First, the results of RT were read visually using single semen samples from 42 consecutive men from infertile couples. In the second step, the results were additionally read using a spectrophotometer. Single semen samples obtained from another 87 consecutive men from infertile couples and 12 men from fertile couples were analysed in this step. Visual analysis, which involved grading against an RT colour chart, revealed a significant positive correlation between the RT results and the quantity and quality of spermatozoa. Azoospermia was diagnosed by grade number 1(dark purple colour), while the highest grades 8–11 (colours dark red to pink) represented a homogeneous group of patients with the highest quality of spermatozoa (>60% motility) and quantity (>60 × 106/ml ejaculate). Normal parameters of spermatozoa, according to WHO, appeared with a high predominance (89–100% of cases) in this group and were present in most of the samples. Thus, the grades 8–11 might represent semen samples with optimal fertility potential. Spectrophotometric analysis of RT revealed that visual characteristics corresponded closely to the values for optical density (OD) of the samples. The OD for normozoospermic samples ranged between 1.48 and 1.84; for oligozoospermia between 1.2 and 1.48; for azoospermia between 0.98 and 1.1. The OD for samples with putative optimal fertility potential ranged between 1.6 and 1.84. RT evaluation using a spectrophotometer may therefore provide a tool for obtaining a wider spectrum of seminological diagnoses than classic microscopic examination, especially for distinguishing men with probable optimal fertility potential.
Effect of defective connective tissue on the formation of aneurysmal‐like structures in the rat testicular artery
Authors: COUTARD MICHÈLE , OSBORNE‐PELLEGRIN MARY
Published: November 2003

Microscopic aneurysmal‐like structures (ALS) develop spontaneously in the convoluted rat testicular artery and have been previously proposed as a model relevant to cerebral aneurysms. The effect of defects in connective tissue fibres on ALS formation was investigated by microscopy using two approaches: (i) the study of the effect of β‐aminopropionitrile (BAPN), an inhibitor of the cross‐linking of elastic and collagen fibres, on the incidence, size and morphology of ALS in spontaneously hypertensive rats (SHR) and their normotensive controls (WKY). The straight spermatic artery was studied for comparison. (ii) The determination of the incidence of spontaneous ALS in Brown Norway (BN) and Long Evans (LE) rats which are highly susceptible (BN) or resistant (LE) to the spontaneous rupture of the arterial internal elastic lamina. (i) BAPN increased the number and size of ALS in SHR and WKY rats and had no effect on the straight spermatic artery and (ii) ALS were more numerous and of greater size in BN than in LE rats. Taken together, these results show that defective connective tissue fibres may favour the formation and induce the enlargement of aneurysmal‐like structures. By analogy, these data suggest that a lack of connective tissue fibre integrity may be of importance in cerebral aneurysm formation and development.
Quantification of tumour‐induced angiogenesis by image analysis
Authors: Iwahana Michio , Nakayama Yutaka , Tanaka Noriko G. , Goryo Masanobu , Okada Kosuke
Published: November 2003

Quantitative techniques for in vivo and in vitro angiogenesis were developed using an image analyser. In the in vivo study, a Millipore chamber filled with mouse sarcoma 180 (S180) cells was transplanted subcutaneously to the dorsal side of a mouse, and the area of neovascularization induced by the tumour cells was quantified by image analysis. Images of vascular networks with poor contrast had their contrast improved by Laplacean transformation. The area of vascular network was 16.9 mm2 in the control group without tumour cells and 44.2 mm2 in the group with tumour cells, demonstrating a significant increase in neovascularized area by tumour cells. In the in vitro study, migration of vascular endothelial cells was induced with conditioned media of S180 cells. Image analysis was used to count automatically the nuclei of migrated endothelial cells, which were stained violet with Giemsa's solution. This automated measurement by image analyser is expected to save labour and time. Checkerboard analysis revealed that the endothelial cell migration induced by S180‐conditioned medium was due to chemotaxis. The quantitation method using an automated image analyser is valuable in evaluating the induction of neovascularization by tumours and the effect of pharmacological agents on tumour angiogenesis in vivo and in vitro
Experimental lung mycotoxicosis in mice induced by Stachybotrys atra
Authors: Nikulin Marjo , Reijula Kari , Jarvis Bruce B. , Hintikka Eeva‐Liisa
Published: November 2003

Stachybotrys atra is often isolated from building materials in houses with moisture problems. Spores of S. atra can contain mycotoxins which may lead to various symptoms in exposed residents in damp houses. The pathogenesis of S. atra‐induced lung diseases has not been elucidated. The purpose of the present study was to investigate lung mycotoxicosis experimentally in mice after an intranasal exposure to spores of S. atra‐fungus. One group of mice received one intranasal injection of spores of a toxic strain of S. atra (1 × 106 spores) and the other group spores of a less toxic strain. Spores of both strains contained spirolactones and spirolactams while the highly toxic strain contained also trichothecene mycotoxins, satratoxins. The spores containing satratoxins caused severe intra‐alveolar, bronchiolar and interstitial inflammation with haemorrhagic exudative processes in the alveolar and bronchiolar lumen. A significant difference was observed in the severity of the lung damage caused by the two strains of S. atra. The spores without satratoxins induced a milder inflammation, so that the toxic compounds of S. atra‐spores are most likely responsible for the severity of the lung injury.
Insulin‐like growth factor‐I modulates monocyte adhesion to EAhy 926 endothelial cells
Authors: MOTANI ALYKHAN , FORSTER LOUISE , TULL SAMANTHA , ÄNGGÅRD ERIK E. , FERNS GORDON A.A.
Published: November 2003

IGF‐I is a ubiquitous growth factor, found in platelets and elaborated by many other cell types. It is thought to be involved in several pathophysiological processes including embryonic development, angiogenesis and wound healing. We report that the adherence of human peripheral blood monocytes to an endothelial cell line (EAhy 926) is inhibited in a dose and time‐dependent manner by pre‐incubating the endothelial cells with IGF‐I (P < 0.001). Monocyte adhesion was inhibited 17.9 ± 1.9% by IGF‐I at a dose of 1000 ng/ml (P < 0.01). In contrast, IGF‐I had no significant effect on monocyte adherence to plastic. The inhibitory effects of IGF‐I were reversed by co‐incubating the endothelial cells with the nitric oxide synthase inhibitor, L‐NAME. These data suggest that the effects of IGF‐I are mediated by the release of nitric oxide from the endothelial cells.
Hepatic extracellular matrix in BALB/c mice infected with Leishmania donovani
Authors: LEITE VIRGINIA H. R. , CROFT SIMON L.
Published: November 2003

The development of the extracellular matrix (ECM) in the liver of the BALB/c mouse infected with Leishmania donovani was observed by histology, immunocytochemistry and electron microscopy at 1, 2, 4, 8, 14 and 20 weeks after infection. Collagen III and proteoglycan were detected in granulomas and in the portal spaces from 4 to 20 weeks after infection. Laminin was not detected in granulomas but was observed in the basement membrane of new small blood vessels in the granulation tissue around the portal spaces from 8 to 20 weeks after infection. The ECM components in the space of Disse showed no changes in distribution throughout the entire period of study. Systemic fibrosis in the hepatic lobule was not evident in the BALB/c mice. This mouse strain does not appear to be an appropriate model to study the role of ECM in chronic visceral leishmaniasis.
The effects of amrinone and glucagon on verapamil‐induced cardiovascular toxicity in anaesthetized rats
Authors: TUNCOK YESIM , APAYDIN SEBNEM , KALKAN SULE , ATES MEHMET , GUVEN HULYA
Published: November 2003

The goal of this study was to compare the effects of glucagon and amrinone on mean arterial pressure (MAP) and heart rate, when used alone and in combination, in an anaesthetized rat model of verapamil toxicity. Rats were anaesthetized and the carotid artery was cannulated for MAP and heart rate measurements. Jugular and femoral veins were cannulated for drug administration. After verapamil infusion (15 mg/kg/h), control animals were given normal saline solution and the other groups received amrinone (0.1 or 0.2 mg/kg/min), glucagon (0.3 mg/kg bolus followed by 0.1 or 0.2 mg/kg/min infusion), glucagon plus amrinone (0.1 mg/kg/min and 0.1 mg/kg/min respectively) or glucagon plus amrinone (0.2 mg/kg/min and 0.1 mg/kg/min respectively). Glucagon (0.2 mg/kg/min) significantly increased MAP when compared to the control group (P < 0.01). The combination of glucagon and amrinone did not produce a synergistic effect for the recovery of MAP. Furthermore, this combination masked the positive effects of glucagon (0.2 mg/kg/min) on MAP.Glucagon (0.2 mg/kg/min) increased the heart rates compared with those of the control group (P < 0.05). Additionally, amrinone (0.1 mg/kg/min) plus glucagon (0.1 mg/kg/min) increased the heart rates (P < 0.05). Finally, glucagon dose dependently recovered MAP. While amrinone depressed MAP in combination with glucagon, it did not alter the positive chronotropic effect of high dose glucagon.
Heparin attenuates bleomycin but not silica‐induced pulmonary fibrosis in mice: possible relationship with involvement of myofibroblasts in bleomycin, and fibroblasts in silica‐induced fibrosis
Authors: PIGUET PIERRE , VAN GWYNETH , GUO JANE
Published: November 2003

Pulmonary fibrosis was elicited in mice or rats by the intra‐tracheal instillation of bleomycin or silica. Daily injections of heparin significantly reduced the collagen deposition in bleomycin, but not in silica, injected mice, as evaluated by the lung hydroxyproline content on day 15 after instillation. Heparin also reduced the bleomycin‐induced morbidity and mortality. Study of the broncho‐alveolar lavage fluid (BAL) detected no significant change in the number of leucocytes or the amount of protein in heparin treated mice. Histologies of bleomycin instilled mice suggested that heparin did reduce the alveolar remodelling but not the alveolitis, evidenced by leucocytic infiltration. As detected by electron microscopy (EM), bleomycin increased the number of leucocytes and platelets within the alveolar capillaries but this was not significantly reduced by heparin. The phenotype of the interstitial cell involved in these two types of pulmonary fibrosis was investigated by immunohistochemistry and EM. While in bleomycin injected animals the interstitial cells had the phenotype of an actin (α‐actin in the rat) and lipid containing interstitial cell, with a poorly developed RE, in silica injected animals in contrast, the interstitial cells were without cytoplasmic actin or lipid but with a markedly developed endoplasmic reticulum (ER). Thus bleomycin and silica induced the growth of two different types of interstitial cells, the myofibroblast and the regular fibroblast, which might be a reason why heparin selectively inhibits bleomycin but not silica‐induced fibrosis.
Haematopoietic progenitor cells transfected with a differentiation antigen show cellular transformation and tumour growt in mice
Authors: HUSS RALF , MYERSON DAVID H. , JOACHIM DEEG H.
Published: November 2003

A stromal cell line, D064, derived from canine bone marrow stroma, was established and differentiates into haematopoietic progenitors under the influence of growth factor signalling. While differentiating, these cells start to express MHC class II molecules (HLA‐DR homologues) on their surface. The transfection of these fibroblast‐like cells with retroviral constructs containing the canine MHC class II DR‐genes (DRA and DRB) induces a change in morphology, alteration of cell cycle progression and tumour formation in nude mice. Transfected cells are smaller than untransfected parental cells and do not require adherence (anchorage dependent growth). The doubling time of untransfected cells was reduced by more than half, as a sign of accelerated cell cycle progression. Injected subcutaneously into nude mice the DR+ transfected cells formed solid tumours, while untransfected cells showed no sign of tumour formation. The transfection‐induced changes were seen only with constructs carrying the open reading frame of DRA plus DRB in the correct orientation and expressing the complete DR‐dimer on the cell surface. Constructs with DRA and DRB in reverse orientation or vectors without any insert did not differ from the parental cells. These observations suggest that mechanisms normally controlling cell cycle and differentiation can be disrupted by the constitutive transcription and expression of differentiation antigens.
Phenotypic variation in hamster bronchial mucous cells induced by different airway irritants
Authors: JAMIL S. , CHRISTENSEN T.G.
Published: November 2003

Chronic mucus hypersecretion (CMH), a common feature of various obstructive pulmonary diseases, is caused by a variety of airway irritants. Bronchial mucous cell metaplasia (MCM), a histological correlate of CMH, can be induced in hamster airways by a number of different irritants. Previous studies with the hamster model suggest that the secretory cell response to different agents is not stereotyped but can vary in the type of mucus glycoconjugate produced. The present ultrastructural study was conducted therefore to provide quantitative evidence of phenotypic variation in mucous cells induced independently by exposure to the metaplastic agents elastase and acid. HPA‐gold lectin cytochemistry revealed an increase in N‐acetyl galactosamine at the cell surface and secretory granules of mucous cells in elastase‐treated vs. acid‐treated animals. Although there was no quantitative difference between the acid‐treated and untreated groups, a difference in the pattern of binding within granules indicated variation in the secretory product. Because mucus glycoconjugates serve as attachment sites for specific pathogens, phenotypically distinct mucous cells may promote differential microbial colonization. In humans therefore, variation in the severity and progression of CMH may be due in part to secretory cell susceptibility and response to different pathogenic stimuli.
Dietary vitamin E supplementation inhibits thrombin‐induced platelet aggregation, but not monocyte adhesiveness, in patients with hypercholesterolaemia
Authors: WILLIAMS J. C. , FORSTER L. A. , TULL S. P. , WONG M. , BEVAN R. J. , FERNS G. A. A.
Published: November 2003

Several recent studies have indicated the possible beneficial effects of antioxidants, specifically vitamin E, in primary and secondary coronary prevention. These studies suggest that a diet enriched in vitamin E is insufficient to have a significant protective effect, whereas supplements, in excess of 200 international units (IU) per day, are efficacious in preventing coronary disease in both men and women. The mechanisms by which vitamin E may exert its protection are uncertain, but, vitamin E is lipophilic and has been shown to inhibit the oxidative modification of low density lipoprotein (LDL), a process thought to be of crucial importance in atherogenesis. We have also previously shown that α‐tocopherol (the biologically most potent isomer of vitamin E) has important direct effects on vascular endothelial and smooth muscle cells. In the present study we have investigated the effects of oral supplements of vitamin E (400 IU per day) on platelet and mononuclear cell function in patients with hypercholesterolaemia. We found that although vitamin E supplementation had no significant effect on mononuclear cell adhesion ex vivo, it had a significant effect on the thrombin‐induced platelet aggregation (P < 0.01; anova): 6 weeks after starting the vitamin E supplements, the mean EC50 for thrombin‐induced aggregation increased 132% (P < 0.05; paired t‐test) compared to treatment with placebo. The effects of vitamin E on platelet function may, in part, explain its anti‐atherogenic properties.