bluuebunny/crossref_metadata_2025 huggingface dataset:
Effectiveness of terbinafine in the eradication ofMicrosporum canisfrom laboratory cats
Authors: Castañón‐Olivares L. R. , Manzano‐Gayosso P. , López‐Martínez R. , De la Rosa‐Velázquez I. A. , Soto‐Reyes‐Solís E.
Published: March 2003

Summary.In this study, the coats of 20 cats with no dermatophytosis lesions were sampled for colonization byMicrosporum canis, which was isolated in 45% of cases. All cats were treated with oral terbinafine for 3 weeks. The drug was administered orally at a daily dosage 8.25 mg kg−1. Mycologic status was assessed at the end of the treatment; and after follow‐up at 2 days, and 4 and 9 weeks. The final evaluation revealed a complete mycologic cure of all cats by obtaining negative cultures, thereby demonstrating that terbinafine is an effective method of eradication of dermatophyte colonization.
One week pulse therapy with itraconazole (200 mg day−1) for onychomycosis. Evaluation of treatment results according to patient background
Authors: Hiruma M. , Matsushita A. , Kobayashi M. , Ogawa H.
Published: March 2003

Summary. We investigated background factors of patients that affect the response of onycomycosis to pulse therapy with itraconazole. The regimen used in this study involved administering 200 mg of itraconazole orally on a daily basis for 1 week as one pulse, which is half of the normally recommended dose. The number of pulses was fixed at one per month, and altered in accordance with improvements in the infected nails and/or the patients' request. Patient background (n = 63) including sex, age, occupation, duration of the disease, site of lesion (fingernail, toenail), number of affected nails, clinical types, severity of thickening, presence or absence of pathogens, the presence or absence of underlying diseases such as adult onset diseases, turbidity of the target nails before therapy and the number and duration of pulses was investigated. The efficacy of the therapy was evaluated after 12 months. A complete cure was judged to have taken place if an entirely healthy nail plate re‐grew in place of the diseased nail, whereas lack of success was defined as the persistence of lesions beyond the designated time period. The average number and duration of pulses were 4.7 ± 3.2 times over 5.6 ± 4.3 months, respectively. The rate of complete cure was 62%. Factors in the patients' background that were judged to have an influence on treatment results were the relative duration of the disease, the number of affected nails, and severity of thickening. With regard to the efficacy of the therapy in terms of the number and duration of pulses, 78% of 23 patients who were given pulse therapy two to six times during a course of 2 to 6 months achieved total cure.
The role of tumour necrosis factor‐α (TNF‐α) and platelet‐activating factor (PAF) interaction on murine candidosis
Authors: Kalkancı A. , Kuştimur S. , Timlioğlu Ö. , Uluoğlu C.
Published: March 2003

Summary.  Tumour necrosis factor‐α (TNF‐α) is related to some other factors in addition to being the essential cytokine of the sepsis which results from Candida infections. In our study, we investigated serum TNF‐α levels, measured by enzyme‐linked immunosorbent assay (ELISA), and platelet‐activating factor (PAF)‐like activity, measured by high‐pressure liquid chromatography (HPLC) of the mice infected with Candida species. The PAF antagonist, ginkgolide BN 52021 was used to evaluate the possible interaction between TNF‐α and PAF. The average TNF‐α levels were found to be 396, 489, 699 and 803 pg ml−1 on the 4th, 5th, 6th and 19th days of Candida albicans infection, respectively (P<0.05). There was no statistically significant difference between the serum TNF‐α levels of the groups infected with other Candida species, such as C. kefyr, C. krusei and C. tropicalis (P>0.05). Serum TNF‐α levels were found to be more significantly different in mice with C. albicans infection that were injected with PAF antagonists on the 6th day (23 pg ml−1). It was therefore thought that PAF antagonists have an inhibitory effect on TNF‐α production. No significant difference was found between PAF levels in the three groups: healthy control mice, C. albicans‐infected mice and C. albicans‐infected mice given PAF antagonists (466 milli‐absorbance unit (mAU), 475 mAU and 329 mAU, respectively). It was noticed that the positive interaction between PAF and TNF‐α was not important after the first 4 days of the infection had passed.
Comparative evaluation of Candi Select test and conventional methods for identification of Candida albicans in routine clinical isolates
Authors: Foongladda S. , Haouharn P. , Sakulmaiwatana P. , Chaiprasert A.
Published: March 2003

Summary. The Candi Select test (Sanofi Diagnostics, Pasteur, Marnes‐La‐Coquette, France) is a new yeast‐selective medium for the identification of Candida albicans in the clinical laboratory. The performance of this test was compared with the conventional methods of chlamydospore formation, germ tube formation and carbohydrate fermentation. Four hundred and twenty clinical yeast isolates from 412 fresh clinical specimens, including 283 C. albicans, 59 C. tropicalis, 39 Trichosporon spp., 19 C. glabrata, 11 Cryptococcus neoformans and 9 other yeasts, were evaluated. Colonies of C. albicans produced a blue‐green colour on the Candi Select media which could be distinguished from the other yeasts with the naked eye within 24–48 h. The sensitivity and specificity of the Candi Select test for the identification of C. albicans were 99.65% and 97.08%, respectively. The blue‐green colonies of C. albicans were easy to identify and recognize in mixed cultures and did not need detailed microscopic examination.
New molecular methods to study gene functions in Candida infections
Authors: Theiß S. , Köhler G. A. , Kretschmar M. , Nichterlein T. , Hacker J.
Published: March 2003

Summary. Candida albicans has become a model system for human pathogenic fungi in clinical research, mainly due to the increasing number of Candida infections. Molecular techniques to study C. albicans virulence properties have been improved over the last few years, despite difficulties in genetic manipulation of this fungus. Some of the recent achievements from our own laboratory or from other groups are described in this article. The molecular analysis of the recently identified ATP‐dependent transporter Mlt1 using the green fluorescent protein (GFP) as reporter for protein localization and the dominant MPAR gene as a selection marker for gene inactivation provides an example for the study of gene functions in C. albicans.
Mycosis on mycosis fungoides: zoophilic dermatophytosis selectively superimposed on pre‐existing cutaneous T‐cell lymphoma (mycosis fungoides) plaques
Authors: Capella G. L. , Altomare G. F.
Published: March 2003

Summary  We report a case of tinea corporis caused by a cattle‐derived strain of Trichophyton mentagrophytes in a 44‐year‐old male affected by cutaneous T‐cell lymphoma (CTCL, so‐called mycosis fungoides). Fungal colonization of glabrous skin was strictly confined within pre‐existing lymphomatous plaques. Either oral itraconazole or griseofulvin, or topical terbinafine were ineffective until the patient, who was treated with systemic retinoids and interferon‐α for his CTCL, was shifted from leucocyte to lymphoblastoid interferon. The hypothesis that a local immunodisturbance could be responsible for the selective superimposition of tinea on CTCL lesions (‘mycosis on mycosis’), and that such an immunodisturbance could be partially corrected by the interferon switch is discussed.
Morphological characterization of modified flower morphology of three novel alloplasmic male sterile carrot sources
Authors: Linke B. , Nothnagel T. , Börner T.
Published: March 2003

Three novel sources of cytoplasmic male sterility (CMS) in carrot are characterized by altered flower phenotypes. Flowers were classified into subtypes, according to stamen (and petal) modification. Flower anatomy was investigated by light microscopy to describe organ modification and to specify the timing when morphology begins to deviate. Early stages of floral development were defined in fertile male flowers of the cultivated carrot according to model plants such as Antirrhinum and Arabidopsis and compared with corresponding stages of the novel cytoplasmic male‐sterile flower types. Early organogenesis was identical in the different CMS types and comparable to corresponding stages of unmodified flowers. The morphology of stamens, and in some cases petals, became different in CMS flowers during early organ differentiation. For each CMS type, the cytoplasm appears to influence organogenesis in a specific way. Although homoeosis is usually considered to be controlled exclusively by specific nuclear genes, a role of cytoplasmic factors is suggested
Characterization and inheritance of partial resistance to downy mildew, Peronospora parasitica, in breeding material of broccoli, Brassica oleracea convar. botrytis var. italica
Authors: Jensen B.D. , Værbak S. , Munk L. , Andersen S. B.
Published: March 2003

Twenty doubled haploid broccoli lines from breeding material were evaluated for resistance to Peronospora parasitica at the seedling stage. All lines supported sporulation of the pathogen but to varying extents and intensities. Partial resistance of the more resistant lines,‘br8’and ‘br9′, reduced conidia production on cotyledons by 50‐70% compared with the most susceptible lines. Inoculation of the two most resistant lines with 13 isolates of different geographical origins revealed that ‘br9′, showed a rather uniform level of resistance to all isolates while ‘br8’showed some isolate specificity Partial resistance was evaluated in six of the broccoli lines in a half diallel set of crosses. Disease assessment of seedlings showed that additive genetic effects explained 45.8% and 31.8% of the total variation of sporulation score and conidia production, respectively This suggests that recurrent selection for partial resistance to P. parasitica in early generation in breds or in populations of broccoli will be efficient to obtain cotyledon resistance.
Changes in genetic diversity during seven cycles of recurrent selection for grainyield in oat, Avena sativa L.
Authors: Koeyer D. L. De , Phillips R. L. , Stuthman D. D.
Published: March 2003

Variability for desirable alleles within elite breeding populations is a requirement for long‐term genetic improvement. Changes in genetic diversity were evaluated in a recurrent selection programme for enhanced grain yield in oat, Avena sativa L., using coefficient of parentage (rp) and restriction fragment length polymorphism (RFLP) based genetic similarity (S) estimates. Both rp and s increased during seven cycles of recurrent selection, indicating a reduction in the level of genetic diversity within the population. The relationship between genetic diversity and agronomic performance was also examined. Genetic similarity measures were poor predictors of the near‐homozygous progeny performance and general and specific combining ability effects for three agronomic traits in the recurrent selection programme. In addition, there was no apparent trend for preferential selection of progeny from either more similar or more diverse parents within a given cycle of selection. The overall reduction in genetic diversity in this population has not affected selection response for grain yield. Variability remaining at important loci or de novo variation are possible explanations for the continued selection progress.
Male sterility in alloplasmic Brassica rapa L. carrying Eruca sativa cytoplasm
Authors: Matsuzawa Y. , Mekiyanon S. , Kaneko Y. , Bang S. W. , Wakui K. , Takahata Y.
Published: March 2003

For the development of a new cytoplasmic male sterility (CMS) system in Brassica rapa (2n= 20, AA), intergeneric hybridization was performed in Eruca sativa (2n= 22, EE)×B. rapa. The original amphihaploid F1 plant (2n= 21, EA) generated via embryo rescue produced a sesquidiploid F1 plant (2n= 31, EAA), from which the alloplasmic F3 plants were generated. In F3, some progenies with malformed anthers were maintained as male‐sterile lines up to the F5 generation. In the F6 and F7 generations, the alloplasmic male‐sterile plants were bred by backcrossing to several B. rapa genotypes and were then classified into the three distinctive types: petaloid, antherless and brown anther, in addition to three intermediate types between them. By southern blot analysis, each plant of the petaloid and antherless types was shown to carry the cytoplasm genome of E. sativa. These male‐sterile plants produced as many viable seeds as the corresponding male‐fertile plants, although their nectary gland development was minimal. Consequently, CMS lines of the petaloid and antherless types with enhanced seed fertility and nectary gland development could provide promising plant materials for F1 seed production in B. rapa.
Changes in expression of HMW glutenins of wheat (Triticum aestivum L.) induced by nitrosoethylurea
Authors: Š Vec M. , Gregová E. , Miklovičová M. , Kraic J.
Published: March 2003

The application of a chemical mutagen, N‐nitroso‐N‐ethylurea, to the grains of wheat (Triticum aestivum L.) cv. ‘Viginta’, provided a mutant line,‘NT‐10′, with an altered composition of high molecular weight (HMW) glutenin subunits. The qualitative mutation was detected in the Glu‐B1 locus by electrophoretic analyses of glutenins. Instead of the HMW glutenin subunits 7 + 9 present in the original genotype, a separate HMW subunit 6 was expressed in the mutant line. The other glutenin and gliadin proteins of the mutant line remained unchanged. The mutant line is also characterized by several changes in morphological and physiological characters—stronger stem, wider leaf, bigger spike and higher grain hardness. This is the first communication of the possibility of changing the composition of high molecular weight subunits of wheat glutenin by means of mutagenesis.
Identification of stable maintainer and fertility restorer lines for ‘Polima’ CMS in Brassica campestris
Authors: Verma J. K. , Sodhi Y. S. , Mukhopadhyay A. , Arumugam N. , Gupta V. , Pental D. , Pradhan A. K.
Published: March 2003

‘Polima’ cytoplasmic male sterility (CMS) was transferred from ‘Polima’ Brassica napus ‘ISN 706’to five different cultivars of Brassica campestris (‘Pusa kalyani’, ‘Pant toria’, ‘Candle’, ‘Tobin’ and ‘ATC 94211′) by repeated backcrossing. It was observed that, while ‘Polima’ CMS manifested complete and stable male sterility in the nuclear backgrounds of ‘Pusa kalyani’, ‘Pant toria’, and ‘Tobin’, the cultivars ‘Candle’ and ‘ATC 94211’possessed the restorer gene for this CMS in the heterozygous condition. An analysis of F1 and F2 generations of ‘Polima’‘Pusa kalyani’×‘Candle’ and ‘Polima’‘Pusa kalyani’×‘ATC 94211’ revealed that restoration is controlled by a single dominant gene. Identification of stable maintainers and restorers of ‘Polima’ CMS could facilitate the development of hybrid varieties in B. campestris.
Concentration effects of dicamba on shoot regeneration in wheat
Authors: Bahieldin A. , Dyer W. E. , Qu R.
Published: March 2003

Shoot formation and regeneration rates of calli derived from immature embryos of three spring wheat varieties were investigated to evaluate the influence on wheat regeneration of different concentrations of dicamba (0.5, 0.1 and 0.02 mg/l) in the regeneration medium. It was found that dicamba concentrations much lower (0.02 and 0.1 mg/l) than the levels reported previously (1 and 0.5 mg/l) induced significantly more shoots per callus in all three varieties, while the enhancement in regeneration rate was variety dependent. Overall, a dicamba concentration of 0.02 mg/l was more favourable for wheat regeneration.
Microsatellite polymorphism in Pisum sativum
Authors: Burstin J. , Deniot G. , Potier J. , Weinachter C. , Aubert G. , Barranger A.
Published: March 2003

Pisum sativum sequences were retrieved from Genbank/EMBL databases and searched for all possible dinucleotide and trinucleotide tandem repeats. One‐hundred and seventy‐one simple sequence repeats (SSRs) were found among 663 sequences. The different dinucleotide or trinucleotide motifs occurred at varying frequencies. CT/AG was the most frequent dinucleotide, and TCT/AGA the most frequent trinucleotide. Forty‐three microsatellite markers were generated from these sequences and used to assess the genetic variability among 12 pea genotypes. Thirty‐one were polymorphic among the genotypes and the average number of variants per marker was 3.6 when considering only polymorphic markers. Overall, the number of variants for a given SSR marker was correlated with the length of the SSR but some 12‐bp long SSRs showed the same degree of polymorphism as longer ones. The groupings resulting from the SSR genotyping among the 12 genotypes gave an interesting insight into the possible origin of one recent cultivar. Database‐derived SSR markers are highly variable. They can provide useful information on the genetic diversity among P. sativum cultivated types.
Detection of chromosomes carrying genes for seed dormancy of wheat using the backcross reciprocal monosomic method
Authors: Miura H. , Sato N. , Kato K. , Amano Y. , McIntosh R. A.
Published: March 2003

Preharvest sprouting (PHS) in wheat represents a major constraint to the production of high‐quality grain. Genetic variation for tolerance to PHS is associated with seed dormancy. The present study was initiated to detect homologous chromosome variation associated with seed dormancy genes in a PHS tolerant cultivar ‘Zenkojikomugi’ (Zen) and ‘Chinese Spring’ (CS) using the backcross reciprocal monosomic method. The most striking effect on variation in seed dormancy was associated with chromosome 3A, and followed by group 4 chromosomes. These chromosomes of Zen increased dormancy compared with the respective CS homologues. Chromosomes 2D and 7D of Zen, and 6A of CS seemed to contribute smaller positive effects on dormancy. Chromosomes 2A of CS and Zen, as well as 3B, were equally effective in enhancing dormancy. The chromosome 3 A factors were independent of the grain‐colour gene R‐A1 because Zen was found to carry the white‐grained R‐A1a allele, which was identical to that of CS, and there was no difference in dormancy between the reciprocal F1 seeds.
A melon genotype with superior competence for regeneration and transformation
Authors: Galperin M. , Patlis L. , Ovadia A. , Wolf D. , Zelcer A. , Kenigsbuch D.
Published: March 2003

Transformation efficiency of melon is low and is still regarded as a challenge. In this paper, the regeneration and transformation response of ‘BU‐21/3′, a newly characterized melon breeding line, is described. The line seems to be superior in this regard to previously evaluated genotypes. Agrobacterium‐mediated delivery of the GUS or GFP reporter genes into cotyledon explants was used to evaluate efficiency of transient and stable transformation. Good transient expression was observed, and stable transformation frequencies of 0.4‐1.5 transgenic shoots per explant were obtained. Transgenic plantlets were transferred to a contained greenhouse as early as 8‐10 weeks after transformation. Transgenic plants are fertile and exhibit a true‐to‐type phenotype. The ‘BU‐21/3’ line may become a useful tool for the facilitation of transgenic breeding in melon.
Cell‐based Systems as an Alternative to Animal Models
Authors: Kues Wa
Published: March 2003

Contents Recent advances in molecular and cellular techniques highlight the potential use of different kinds of somatic, as well as embryonic and adult stem cell types in efficient and rapid in vitro test systems. The rapidly progressing genome sequencing projects will permit studies based on sequence‐defined cell lines. Thus a combination of genomic data, molecular tools and appropriate cells together with a spectrum of high throughput analysis methods, such as cDNA arrays, two‐hybrid analysis, Maldi‐TOF spectrometry and protein chips will produce powerful test systems for target gene, as well as target protein identification and for toxicological and pharmacogenetic testing. These studies will be performed first in vitro but subsequently cellular test systems will additionally be required. The modern cell factory allows the researcher to answer specific questions about the role of genes, proteins and metabolites, as well as several other factors. The employment of embryonic stem cells and sophisticated methods to direct their differentiation towards desired mature cell types or differentiation states, will allow investigation into the molecular switches controlling embryonic development, differentiation, tumorigenicity and teratogenicity. The topic of this article is whether cell‐based systems could be an alternative to the usage of animal, or more precisely whether cell‐based systems could probably be the better alternative?
First Identification of Caldesmon Transcripts in Bovine Oviduct Epithelial Cells in vitro by means of an RNA Differential Display Technique Examining Culture‐induced Expression Changes
Authors: Einspanier R , Bieser B , Reischl J , Prelle K
Published: March 2003

Innovative molecular biology techniques enable the quick evaluation of distinct gene expression pattern of cells or tissues. Hitherto, a cell‐type‐specific behaviour has been difficult to evaluate. In addition to standard morphological and immunological criteria in this study the expression of in vitro‐cultured bovine oviduct epithelial cells (BOECs) was compared with fresh cells by RNA arbitrarily primed polymerase chain reaction (RAP‐PCR). The cultured cells showed mitotic activity during the whole culture period (6 days) until they had reached about 80% confluency. Remarkable results of a random transcript screening using fresh versus cultured BOECs were reported, in which a single PCR product appeared during culture, but was absent in fresh cells. Sequence analysis of the culture‐induced 522 bp fragment revealed a high homology (87%) to caldesmon (CaD) of various species and a 92% homology to a short cDNA fragment of a bovine non‐muscle CaD. Specific, cross‐species PCR primers were used to elongate this partial sequence (1036 bp). This resulting cDNA showed an open reading frame and was identified as a bovine non‐muscle CaD isoform. When compared with human non‐muscle CaDs (89% homology) a deletion of 2 codons was observed. According to sequential culture experiments, CaD expression was not found in fresh BOECs but specific transcripts appeared within 48–113 h under specific culture conditions. It is likely that augmented CaD expression in cultured BOECs may reflect the cell effort adapting to specific culture conditions. The hypothesis that increased CaD levels could be important to facilitate adherence and spreading by formation of new stress fibres can be favoured. This first identification of caldesmon expression being specifically induced during in vitro culture demonstrates the potential of RAP‐PCR for the analysis and validation of cell culture techniques.
Comparison of External and Internal Pelvic Measurements of Belgian Blue Cattle from Sample Herds in Belgium and the United Kingdom
Authors: Murray RD , Cartwright TA , Downham DY , Murray MA , De Kruif A
Published: March 2003

Measurements describing pelvic conformation of pedigree Belgian Blue cows were obtained from a sample of nine herds in Flanders, Belgium, comprising 111 adult breeding cows, and from 11 herds in the United Kingdom comprising 108 similar cows. All herds in the Belgian sample (111 cows) managed parturition by elective caesarian section as did seven herds (56 cows) in the UK sample; the remainder of UK herds (52 cows) allowed cows the opportunity to calve naturally per vagina before resorting to caesarian section. The data described the external and internal measurements, and pelvic area characteristic of this breed, irrespective of the different selection pressures applied over a number of years to this breed through variation in farm management and market forces present in either country. From these data, generalized linear models were constructed to predict pelvic area; they correctly identified cows with either small or large pelvic areas. There were no significant differences in pelvic conformation between cattle bred either in Belgium or the UK, although those cows bred in herds where natural calving was allowed to take place had significantly larger internal pelvic height (p < 0.05) and area (p < 0.05) than other Belgian Blue cows. The correlation coefficients between internal pelvic height and width and external pelvic measurements were significant (p < 0.001). These results might facilitate the selection of breeding cows with larger pelvic area so that a higher proportion of cows can calve naturally than currently occurs. However, selection for other traits such as relatively low birth weight combined with higher weaning weight should be carried out at the same time.
Vitamin A and β‐Carotene Levels in Plasma, Corpus Luteum and Follicular Fluid of Cyclic and Pregnant Cattle
Authors: Haliloglu S , Baspinar N , Serpek B , Erdem H , Bulut Z
Published: March 2003

Contents

This study was carried out to examine the relationship between the corpus luteum (CL) weight, CL and follicle diameters and progesterone, β‐carotene and vitamin A levels in reproductive organs of cattle obtained from the slaughterhouse. The β‐carotene and vitamin A levels were determined in plasma, CL and follicular fluid (FF) using a spectrophotometric method at different stages of the oestrous cycle (n=40) and at 3–6 months of pregnancy (n=10). The diameters of the CL and follicle were measured using ultrasonography. Plasma progesterone concentrations were determined by an enzyme immunoassay method. The vitamin A levels of the plasma, CL and FF were not related to each other. The highest plasma vitamin A levels were observed in the proestrus and oestrus, at which periods follicular activity dominates. The vitamin A levels in the CL and FF were negatively related to the weight and diameter of the CL and the diameter of follicle, respectively. In contrast to vitamin A, β‐carotene concentrations of plasma, CL and FF were significantly correlated with each other. The highest β‐carotene levels in the plasma, CL and FF were found during pregnancy when there is maximal luteal function, and the β‐carotene level of the CL was significantly correlated with the weight and diameter of CL. Furthermore, the intrafollicular β‐carotene level was negatively correlated with the follicle diameter. There was a positive correlation between plasma progesterone level and the weight and diameter of the CL, but a negative correlation between plasma progesterone level and follicle diameter. Moreover, plasma, FF and CL β‐carotene levels were positively correlated with plasma progesterone levels. This study revealed that β‐carotene levels in the plasma, CL and FF were influenced by the stage of the oestrous cycle or the pregnancy and were related to bovine luteal function without depending on vitamin A.
Function of the Cumulus Oophorus Before and During Mammalian Fertilization
Authors: Van Soom A , Tanghe S , De Pauw I , Maes D , De Kruif A
Published: March 2003

Contents

Fertilization encompasses a series of different steps which have to be performed in a well‐orchestrated way to create a new individual. They include sperm capacitation, sperm binding and penetration of the zona pellucida, traversing the perivitelline space, binding and fusion with the oolemma, activation of the oocyte and decondensation of the sperm head to form the male pronucleus. In most mammalian species, cumulus cells surround the oocyte at the time of fertilization. Removal of the cumulus oophorus at this point of time often leads to a drop in fertilization rates. It is not yet known how cumulus cells interact with the oocyte or with spermatozoa to promote fertilization. There are different possibilities:

cumulus cells cause mechanical entrapment of spermatozoa and guide hyperactivated spermatozoa towards the oocyte, while preventing abnormal spermatozoa to enter the cumulus matrix;

cumulus cells create a micro‐environment for the spermatozoa which favours their capacitation and penetration into the oocyte;

cumulus cells prevent changes in the oocyte which are unfavourable for normal fertilization; these changes can be located in the zona pellucida or in the cytoplasm.

In this review, studies in several species are listed to prove the importance of these three cumulus cell functions and the current lines of research are highlighted. Moreover, different ways to improve in vitro fertilization of bovine cumulus‐denuded oocytes are discussed.
Effect of Albumin and Polyvinyl Alcohol on the Vitality, Motility and Acrosomal Integrity of Canine Spermatozoa Incubated in vitro
Authors: Risopatrón J , Catalán S , Miska W , Schill W‐B , Sánchez R
Published: March 2003

Contents

Sperm culture media used for in vitro fertilization (IVF) procedures are important factors concerning the viability, motility and acrosomal integrity of spermatozoa. The aim of this study was to investigate the effects of three different sperm diluting media, tissue culture medium (TCM‐199), sperm culture medium (Sp‐TALP) and human tubular fluid (HTF) supplemented with varying concentrations of bovine serum albumin (1, 4 and 6%) or polyvinyl alcohol (0.8%) on the acrosomal integrity, motility and viability of canine spermatozoa. Ejaculates collected from four dogs were diluted in all media and spermatozoa were separated from seminal plasma by the swim‐up technique. Sperm progressive motility was assessed using a phase contrast microscope. Viability and acrosomal integrity were evaluated using a dual stain technique (Giemsa–Trypan blue). The results demonstrated that the number of live canine spermatozoa was similar in culture media supplemented or not supplemented with macromolecules. A minimal concentration of albumin (1%) in the three media showed similar effects on vitality, motility and acrosomal integrity, as had higher concentrations (4 and 6%). The percentage of acrosome‐intact spermatozoa was markedly higher after HTF (94.1%) than after TCM‐199 (70.1%) or Sp‐TALP (71.0%) without supplementation. It is concluded that serum bovine albumin, irrespective of the concentration, preserved sperm viability and function, and HTF is the most suitable medium for preserving the acrosome in canine spermatozoa prepared for in vitro manipulation through short incubation.
Is depression normal in human beings? A critique of the evolutionary perspective
Authors: McLoughlin Garry
Published: March 2003

ABSTRACT:

To the evolutionary biologist human beings at every stage of their development represent ‘compromises’ in their continual adaptation to their changing environments. Using a neo‐Darwinian perspective, evolutionary psychiatrists such as Randolph Nesse (Professor of Psychiatry at the University of Michigan) argue that while natural selection does not shape disease itself, it does shape human traits and therefore vulnerability to disease. Accordingly, for him, depression is a human emotion which may represent a surviving positive response and is therefore not always pathological. This critique examines Nesse's principal arguments and reveals a number of weaknesses in those arguments. The article concludes with a review of the therapeutic and preventive implications of his evolutionary perspective on depressive states as well as some implications for mental health nurses.
The eosinophil leucocyte, a phenotypic marker of resistance to nematode parasites, is associated with calm behaviour in sheep
Authors: Hohenhaus MA , Josey MJ , Dobson C , Outteridge PM
Published: March 2003

The hypothesis that behaviour in sheep is influenced by resistance to infections with Trichostrongylus colubriformis and Haemonchus contortus was explored. Sheep were assessed phenotypically as resistant and susceptible in four ways and thereafter, the effect of resistance on several behavioural traits was measured in an arena test. The behavioural parameters recorded for each sheep were: approach/avoidance distance; travel; number of moves; and spread. Four phenotypic groups of sheep were set up each with two subgroups: ovine lymphocyte antigen (OLA) type (subgroups SY1a type vs other types); blood eosinophil leucocyte counts (high vs low); T. colubriformis and H. contortus serum antibodies (high vs low titres) and faecal egg count (FEC) (high vs low).

Only the behavioural comparison between sheep with high versus low eosinophil leucocyte count showed consistent differences that were statistically significant, although other comparisons favoured associations between OLA type SY1a, low FEC and reduced approach/avoidance distance and/or locomotor behaviour. The eosinophil leucocyte association was established on two non‐overlapping test populations of sheep, with mean eosinophil leucocyte counts of 3.19 × 105/mL blood for high and 0.83 × 105/mL blood for low subgroups; results were reproduced on five separate occasions over 6 weeks to detect differences as approach/avoidance distance (P < 0.01), travel (P < 0.01), number of moves (P < 0.01), and spread (P < 0.05).

High eosinophilia correlated with low FEC for the infected sheep, which identified sheep that were resistant to parasites. We concluded, therefore, that the resistant sheep were more at ease with the environment of the test than were the sheep with low eosinophil leucocyte counts. The strong association between high eosinophil leucocyte counts and behaviour is consistent with other reports which suggest that retention of normal levels of circulating eosinophil leucocytes is associated with resistance to stress.
Expression of biologically active recombinant porcine GM‐CSF by baculovirus gene expression system
Authors: Inumaru S , Kokuho T , Denham S , Denyer MS , Momotani E , Kitamura S , Corteyn A , Brookes S , Parkhouse Rme , Takamatsu H
Published: March 2003

The full length porcine granulocyte/macrophage colony stimulating factor (GM‐CSF) cDNA, including secretion signal peptide coding region was recloned into baculovirus transfer vector pAcYM1. The vector was then transfected with Autographica californica nuclear polyhedrosis virus (AcNPV) DNA into SF21AE cells and the recombinant virus AcPGM was recovered. Recombinant porcine GM‐CSF (rpGM‐CSF) was obtained from the serum‐free culture medium of Tn5 cells infected with the AcPGM virus, and was shown to be a glycosylated 21 kDa protein as confirmed by tunicamycin treatment and [3H]‐glucosamine uptake. The biological activities of rpGM‐CSF in AcPGM‐infected cell culture supernatants were demonstrated by porcine bone marrow cell proliferation and haematopoietic cell colony formation assays. The use of rpGM‐CSF enabled us to culture porcine monocytes/macrophage and dendritic‐like cells, derived from either porcine bone marrow or peripheral blood, for up to 4 months.
Modulation of HLA class I antigen and ICAM‐2 on endothelial cells after in vitro infection with human cytomegalovirus
Authors: Shahgasempour Shapour , Woodroffe Salwa B , Garnett Helen M
Published: March 2003

The aim of the present work is to investigate the expression of HLA class I antigen and intercellular adhesion molecule‐2 (ICAM‐2) on the surface of human umbilical cord endothelial cells (HUVEC) after infection with human cytomegalovirus (HCMV). The expression of HLA class I antigen and ICAM‐2 were determined (using antibodies against HLA class I antigen and ICAM‐2) by attachment inhibition assay and flow cytometric analysis. Attachment inhibition assay demonstrated that HCMV increased the expression of HLA class I antigen. This was confirmed by flow cytometric analysis, which showed an increase in HLA class I antigen expression on HCMV‐infected HUVEC. The results of the expression of ICAM‐2 using attachment inhibition assay revealed that ICAM‐2 is involved significantly in the increased adhesion of T lymphocytes to HCMV‐infected HUVEC. However, flow cytometric analysis revealed that there were no changes in the expression of ICAM‐2 on HCMV‐infected HUVEC. One possible explanation for this is that HCMV induces the activation of ICAM‐2 on the surface of HCMV‐infected endothelial cells without affecting its expression.
Visualizing lymphocyte recognition
Authors: Wülfing Christoph , Chien Yueh‐Hsiu , Davis Mark M
Published: March 2003

Studies of T cell recognition have entered new territory now that some of the basic issues of genetics, biochemistry and structure have been addressed, at least in outline form. In the present work, the focus is on a new aspect of T cell recognition that goes beyond classical biochemistry to ask, ‘how do TCR and other cell surface molecules cooperate to initiate and control recognition?’
Dendritic cell origins: Puzzles and paradoxes
Authors: Vandenabeele Stéphane , Wu Li
Published: March 2003

In the present review, a series of studies on the origins of dendritic cells of mice and humans are summarized. Several subsets of mature dendritic cells found in vivo are described and these may correspond to distinct lineages. There is evidence that some dendritic cells are myeloid‐derived and that others are lymphoid‐derived. The different ways of generating dendritic cells are examined and an attempt to reconcile the differences seen using mouse and human culture models is made. The particular case of Langerhans cells is discussed and an historical overview of the biology of the plasmacytoid T cells, which may represent a distinct ‘lymphoid‐related’ dendritic cell lineage, is given. It is concluded that three or four different pathways lead to the development of different subtypes of dendritic cells.
Nuclear and cytoskeletal translocation and localization of heterotrimeric G‐proteins
Authors: Willard Francis S , Crouch Michael F
Published: March 2003

Heterotrimeric GTP‐binding proteins (G‐proteins) are involved in a diverse array of signalling pathways. They are generally thought to be membrane‐bound proteins, which disassociate on receptor activation and binding of GTP. A model to explain this has been proposed, which is often described as ‘the G‐protein cycle’. The ‘G‐protein cycle’ is discussed in the present paper in relation to evidence that now exists regarding the non‐ membranous localization of G‐proteins. Specifically, the experimental evidence demonstrating association of G‐proteins with the cytoskeleton and the nucleus, and the mechanisms by which G‐proteins translocate to these sites are reviewed. Furthermore, the possible effector pathways and the physiological function of G‐proteins at these sites are discussed.
Computer‐assisted analysis of molecular mimicry between human papillomavirus 16 E7 oncoprotein and human protein sequences
Authors: Natale Costanzo , Giannini Teresa , Lucchese Alberta , Kanduc Darja
Published: March 2003

The immunology of human papillomavirus (HPV) infections has peculiar characteristics. The long latency for cervical cancer development after primary viral infection suggests mechanisms that may aid the virus in avoiding the host immunosurveillance and establishing persistent infections. In order to understand whether molecular mimicry phenomena might explain the ability of HPV to avoid a protective immune response by the host cell, sequence similarity between HPV16 E7 oncoprotein and human self‐proteins was examined by computer‐assisted analysis. Data were obtained showing that the HPV16 E7 protein has high and widespread similarity to several human proteins involved in a number of critical regulatory processes. In addition, multiple identical and different E7 peptide motifs are present in the same human protein. Thus, sharing of common motifs between viral oncoproteins and molecules of normal cells may be one cause underlying the scarce immunogenicity of HPV infections. The hypothesis is advanced that synthetic peptides harbouring viral motifs not and/or scarcely represented in the host's cellular proteins may represent a valuable immunotherapeutic approach for cervical cancer treatment.
Induction of CD8+ T‐lymphocyte responses to a secreted antigen of Mycobacterium tuberculosis by an attenuated vaccinia virus
Authors: Feng Carl G , Blanchard Tom J , Smith Geoffrey L , Hill Adrian VS , Britton Warwick J
Published: March 2003

Protective immunity against Mycobacterium tuberculosis infection requires the activation of mycobacterium‐specific CD8+ T cells, as well as CD4+ T cells. Therefore, optimizing strategies that stimulate CD8+ T cells recognizing dominant mycobacterial antigens, including secreted proteins, may lead to the development of more effective vaccines against tuberculosis. To generate a viral vaccine that is safe in humans, the early secreted protein, MPT64, was expressed in the attenuated vaccinia virus (VV) strain, modified vaccinia virus Ankara (MVA‐64). The immunogenicity of MVA‐64 was compared with that of the Western Reserve strain of VV (VVWR‐64). The replication‐defective MVA‐64 was as efficient as VVWR‐64 in inducing specific antibodies and cytolytic T‐cell responses to a defined H‐2‐Db‐restricted epitope on MTP‐64. In addition, priming with MPT64‐expressing plasmid DNA (DNA‐64), and boosting with either MVA‐64 or VVWR‐64, markedly enhanced MPT64‐specific cytolytic and IFN‐γ‐producing CD8+ T‐cell responses. These findings suggest that MVA may be a suitable vaccine carrier for stimulating mycobacterium‐specific CD8+ T‐cell responses and may be particularly relevant for developing vaccines for use in regions endemic for tuberculosis and HIV infection.
Activation‐induced cell death: The controversial role of Fas and Fas ligand in immune privilege and tumour counterattack
Authors: Maher Stephen , Toomey Deirdre , Condron Claire , Bouchier‐Hayes David
Published: March 2003

Activation‐induced cell death (AICD) is the process by which cells undergo apoptosis in a controlled manner through the interaction of a death factor and its receptor. Programmed cell death can be induced by a number of physiological and pathological factors including Fas (CD95)–Fas ligand (FasL/CD95L) interaction, tumour necrosis factor (TNF), ceramide, and reactive oxygen species (ROS). Fas is a 48‐kDa type I transmembrane protein that belongs to the TNF/nerve growth factor receptor superfamily. FasL is a 40‐kDa type II transmembrane protein that belongs to the TNF superfamily. The interaction of Fas with FasL results in a series of signal transductions which initiate apoptosis. The induction of apoptosis in this manner is termed AICD. Activation‐induced cell death and Fas–FasL interactions have been shown to play significant roles in immune system homeostasis. In this review the involvement of Fas and Fas ligand in cell death, with particular reference to the T cell, and the mechanism(s) by which they induce cell death is described. The role of AICD in immune system homeostasis and the controversy surrounding the role of FasL in immune privilege, inflammation, and so‐called tumour counterattack is also discussed.
Proteasome inhibitor MG‐132 enhances the expression of interleukin‐6 in human umbilical vein endothelial cells: Involvement of MAP/ERK kinase
Authors: Shibata Takeo , Imaizumi Tadaatsu , Tamo Wakako , Matsumiya Tomoh , Kumagai Mika , Cui Xue‐Fan , Yoshida Hidemi , Takaya Shun‐Ichi , Fukuda Ikuo , Satoh Kei
Published: March 2003

Interleukin‐6 (IL‐6) is a multifunctional cytokine that plays an important role in inflammatory reactions. We have addressed the possible regulation of IL‐6 expression by the ubiquitin‐protease system in human umbilical vein endothelial cells. Cultured endothelial cells were treated with MG‐132, a protease inhibitor, and the levels of IL‐6 mRNA and protein were measured by reverse transcription‐PCR and ELISA. MG‐132 increased the expression of IL‐6 mRNA and protein; and this effect was abolished by the pretreatment of the cells with U0126, an inhibitor of MAP or ERK kinases (MEK1/2). MG‐132 treatment was also found to enhance the level of phosphorylated MEK1/2. Treatment of the cells with actinomycin D inhibited IL‐6 expression in response to MG‐132, suggesting the transcriptional upregulation of IL‐6 under proteasomal inhibition. We conclude that a protease inhibitor MG‐132 upregulates IL‐6 expression in vascular endothelial cells, at least in part, through the activation of MEK1/2.
Pain in children: Qualitative research of Finnish school‐aged children’s experiences of pain in hospital
Authors: Pölkki Tarja , Pietilä Anna‐Maija , Rissanen Lea
Published: March 2003

The purpose of the study was to describe school‐aged children’s experiences of pain in the hospital. Data were collected from 20 children aged between seven and 11 years of age who were inpatients in the pediatric wards of the University Hospital, Oulu, Finland. Each of the children wrote of their experiences of pain in the hospital, and they were interviewed on the basis of this information. The data were analyzed inductively using content analysis. On the basis of the analysis, the situations that caused pain to all children in the hospital were found to be procedures connected with needles. The children described their experiences of pain as both physiological (e.g. poking, stinging, aching) and psychological feelings (unpleasant feelings, terror, anxiety, and fear). In addition, methods of relieving pain (coping mechanisms, help from the medical staff, parental presence, and previous experiences of similar situations) were acknowledged. The results indicate that school‐aged children (aged 7–11 years) are able to describe their pain experiences, which should be considered in assessment and treatment of children’s pain in nursing practice.
Palliative care at home: Stress for nurses in urban and rural New South Wales, Australia
Authors: Wilkes Lesley M , Beale Barbara
Published: March 2003

Nurses often experience positive and negative dimensions of caring for dying clients and their families. This project aimed to compare stress experienced by urban and rural community nurses working with palliative‐care clients in the home. Participants included five nurses working in rural Australia and seven nurses working in an urban area. Transcribed data from unstructured audio‐taped interviews were coded for common and contrasting themes, and a comparison was made of the stress experienced by the two groups of nurses. The major themes were role conflict and definition, family dynamics, time and workload. For both groups of nurses, the impact of family relationships and role conflict within the community impacted significantly to the stress they experienced. Debriefing opportunities for nurses to discuss stress, including educational and support sessions, is an essential component of best practice. Rural nurses had the additional stress of trying to provide a 24 h service over vast distances with a lack of financial resources.
Factors associated with the coping of parents with a child in psychiatric inpatient care
Authors: Puotiniemi Tiina , Kyngäs Helvi , Nikkonen Merja
Published: March 2003

The purpose of this study was to establish the parental coping' factors associated with having a child in psychiatric inpatient care. The data were collected from 19 hospitals with child psychiatry units. At the time of data collection, all parents of children in psychiatric inpatient care in these hospitals were recruited. The method of data collection was a questionnaire (n = 79). The data were analysed with the Statistical Package for the Social Sciences (SPSS) for Windows statistical software. The connections between variables were studied with cross‐tabulation, and the χ2 test was used to determine significance. Changes in internal and external family relationships and matters related to the upbringing of the child with mental problems statistically correlated significantly with parental coping (P < 0.001). Problem‐oriented and emotionally‐oriented coping strategies, skills and palliative strategies correlated significantly with parental coping (P < 0.001). Emotional support, support for the care and upbringing of the child in inpatient care, and love and acceptance also had statistically significant associations with parental coping (P < 0.001).