bluuebunny/crossref_metadata_2025 huggingface dataset:
A Population‐Based Approach to Pediculosis Management
Authors: Monsen Karen A. , Keller Linda Olson
Published: March 2003

Pediculosis is a population‐based problem that is a priority concern for many communities. Public health nurses in a county health department collaborated with epidemiologists, nursing students, and faculty to design and implement an effective population‐based pediculosis management project. The focus of the project was the development of pediculosis treatment and prevention guidelines based on recognized best practices that were acceptable to both epidemiologists and practicing public health nurses. Public health nurses disseminated these guidelines to community providers and reinforced their use through consultation and educational sessions. Two critical changes occurred as a result of the project. First, community providers significantly changed their recommendations for the treatment of pediculosis after nursing intervention. Second, public health nurses increased their population‐based practice skills, continued to use those skills to address pediculosis, and extended those skills to additional population‐based initiatives.
Nicotinic acetylcholine receptor mRNA in dorsal root ganglion neurons
Authors: Boyd R. Thomas , Jacob Michele H. , McEachern Adrienne E. , Caron Suzanne , Berg Darwin K.
Published: March 2003

Nicotinic acetylcholine receptor (AChR) genes are expressed in subpopulations of chick dorsal root ganglion (DRG) neurons. In 18‐day embryonic ganglia, 19% of the neurons have material homologous to neuronal AChR α3 gene mRNA, and 8% have material homologous to α4 mRNA as seen with in situ hybridization. RNAase protection experiments confirm that DRG RNA contains α3 and α4 transcripts, and Northern blot analysis establishes the size of the transcripts as being 3.5 and 3.3 kb, respectively. The proportion of DRG neurons containing α3 mRNA does not decline up through 1 year post‐hatch, indicating that α3 gene expression is not a developmentally transient event in the ganglion. An antiAChR monoclonal antibody detects cross‐reacting material in 16% of the DRG neurons from 18‐day embryos, indicating that AChR mRNA is translated into protein. Electrophysiological measurements confirm the presence of functional AChRs on DRG neurons freshly isolated from 18‐day embryos: 24% of the neurons have substantial ACh sensitivities, whereas another 23% have small but detectable responses. Staining dorsal root ganglion sections with an anticholine acetyltransferase antiserum reveals cross‐reactive material localized in axons in the ganglion; no evidence suggests the presence of cholinergic synaptic structures or AChR clusters on neuronal somata in the ganglion. It is possible that AChRs on DRG neurons participate in a diffuse form of transmission between the cholinergic fibers and a subpopulation of neuronal somata in the ganglion. Alternatively, AChRs on the somata may represent an ectopic distribution of receptors whose primary function is at the terminals of central or peripheral DRG processes.
Influence of plant lecithin partly replacing rumen‐protected fat on digestion, metabolic traits and performance of dairy cows
Authors: Wettstein H.‐R. , Quarella Forni M. G. , Kreuzer M. , Sutter F.
Published: March 2003

Nutrient digestibility, metabolic effects and performance were evaluated in lactating cows (n = 6 × 6) receiving diets containing one of four different lecithins (raw, deoiled and deoiled/partially hydrolysed soy lecithin, raw canola lecithin) or soybean oil in combination with Ca soaps of palm oil fatty acids in a ratio of 1 : 3. A diet supplemented only with Ca soaps served as a control. The complete diets consisted of forage and concentrate (0.62 : 0.38) and total lipid supplementation was 30 g fatty acids/kg dietary dry matter. Lecithins had no effect on milk yield and milk composition. Apparent digestibility of dry matter, organic matter and gross energy were highest in the control diet and lowest in the soybean oil diet (p < 0.1) with intermediate values for the lecithin‐supplemented diets. Soybean oil particularly reduced fibre digestibility (significant with hemicellulose), whereas this effect was reduced or even missing when lecithins, particularly raw canola lecithin and deoiled/hydrolysed soy lecithin, were used. The digestibility of total fatty acids was not affected by the treatments. The effects were also not significant for blood plasma traits except for insulin which was significantly higher with raw than with deoiled/hydrolysed soy lecithin. In conclusion, lecithins, particularly those technologically modified or of a low polyenoic fatty acid content, might be preferred to oils when used as an energy source whereas they failed to improve digestibility of rumen‐protected fats.
Ouabain‐sensitive respiration and protein synthesis in duodenal mucosa and liver in rats fed increasing levels of pea fibre or protein and housed in 18 or 28°C environments
Authors: Zhao Xin‐Quan , Jørgensen H. , Gabert V. M.
Published: March 2003

Seventy‐two Wistar rats were used in two studies to investigate the effect of environmental temperature (18 or 28°C), and increasing levels of dietary fibre (low, 68 g/kg dry matter (DM); medium 110 g/kg DM; high, 157 g/kg DM) and protein (low, 91 g/kg DM; medium, 171 g/kg DM; high, 262 g/kg DM) on respiration attributable to Na+,K+‐ATPase activity and protein synthesis in duodenal mucosa and liver of rats. In vitro O2 consumption in tissues was measured polarographically using a Clark‐style YSI biological O2 monitor. Whole‐body O2 consumption was measured with two open‐circuit respiration chambers. Whole‐body O2 consumption was higher (p < 0.05) at 18°C than at 28°C. Rats fed the low protein diet had significantly higher (p < 0.05) whole‐body O2 consumption than those fed the medium or high protein diet. Compared with 28°C, the environmental temperature of 18°C caused an increase (p < 0.05) in total O2 consumption and O2 consumption attributable to Na+,K+‐ATPase activity in duodenal mucosa. There was no effect (p > 0.05) of environmental temperature on total O2 consumption, Na+,K+‐ATPase activity attributable to protein synthesis dependent on O2 consumption in the liver. Total O2 consumption and O2 consumption attributable to Na+,K+‐ATPase activity increased (p < 0.05) in duodenal mucosa in rats fed the low level of dietary fibre compared with rats fed the medium level of dietary fibre. In vitro O2 consumption determined in duodenal mucosa and in liver did not always correspond to whole‐body O2 consumption. This may indicate that respiration in the duodenum and liver adapts differently and may not reflect changes in whole‐body respiration in response to dietary modification and changes in thermal environment.
First isolation of Trichophyton raubitschekii (syn. T. rubrum) in Europe
Authors: Tietz H.‐J. , Hopp M. , Gräser Y
Published: March 2003

Summary. Trichophyton raubitschekii is a rare dermatophyte which belongs to the Trichophyton rubrum species complex. Since 1981, only a few cases of dermatophytosis due to this anthropophilic causative agent were published. In this paper the authors report the first cases of Tinea corporis caused by Trichophyton raubitschekii (syn. T. rubrum) in Europe. The patients, one immigrant from Ghana and three from Cameroon, had typical lesions of tinea corporis. Four strains were isolated and characterized by conventional and molecular methods. On morphological and physiological grounds the isolates were identified as T. raubitschekii by the following phenotypical features: (1) velvety colony texture; (2) brown pigment; (3) abundant macroconidia and (4) positive urease activity. Molecular diagnostics were performed by single strand conformation polymorphism (SSCP) and sequence analysis of the ATPase9 intron of the mitochondrial (mt) DNA and the internal transcribed spacer (ITS) region of the rDNA, respectively. The ITS sequences and SSCP patterns of the ATPase9 intron were found to be identical among the four strains and also when compared to reference strains of T. rubrum. As shown in the present paper, T. raubitschekii is genetically identical to T. rubrum but differs in some phenotypical characteristics. Since misidentification with other dark‐coloured dermatophyte variants is possible, medical mycologists should bear in mind the special morphological characteristics of T. raubitschekii (syn. T. rubrum) for future identifications.
A retrospective study of canine and feline cutaneous vasculitis
Authors: Nichols Patrick R. , Morris Daniel O. , Beale Karin M.
Published: March 2003

Twenty‐one cases of cutaneous vasculitis in small animals (dogs and cats) were reviewed, and cases were divided by clinical signs into five groups. An attempt was made to correlate clinical types of vasculitis with histological inflammatory patterns, response to therapeutic drugs and prognosis. Greater than 50% of the cases were idiopathic, whereas five were induced by rabies vaccine, two were associated with hypersensitivity to beef, one was associated with lymphosarcoma and two were associated with the administration of oral drugs (ivermectin and itraconazole). Only the cases of rabies vaccine‐induced vasculitis in dogs had a consistent histological inflammatory pattern (mononuclear/nonleukocytoclastic) and were responsive to combination therapy with prednisone and pentoxifylline, or to prednisone alone. Most cases with neutrophilic or neutrophilic/eosinophilic inflammatory patterns histologically did not respond to pentoxifylline, but responded to sulfone/sulfonamide drugs, prednisone, or a combination of the two.
Synthesis, processing and export of cytoplasmic endo‐β‐1,4‐xylanase from barley aleurone during germination
Authors: Caspers Martien P. M. , Lok Finn , Sinjorgo Karin M. C. , Van Zeijl Mieke J. , Nielsen Kirsten A. , Cameron‐Mills Verena
Published: March 2003

We have identified the major endo‐β‐1,4‐xylanase (XYN‐1) in the aleurone of germinating barley grain, and show that it is expressed as a precursor of Mr 61 500 with both N‐ and C‐terminal propeptides. XYN‐1 is synthesized as an inactive enzyme in the cytoplasm, and only becomes active at a late stage of germination when the aleurone ceases to secrete hydrolases. A series of processing steps, mediated in part by aleurone cysteine endoproteases, yields a mature active enzyme of Mr 34 000. Processing and extracellular release of the mature enzyme coincide with the programmed cell death (PCD)‐regulated disintegration of aleurone cells. We discuss the significance of delayed aleurone cell‐wall degradation by endoxylanases in relation to the secretory capacity of the aleurone, and propose a novel role for aleurone PCD in facilitating the export of hydrolases.
Runaway cell death, but not basal disease resistance, in lsd1 is SA‐ and NIM1/NPR1‐dependent
Authors: Aviv Daniel H. , Rustérucci Christine , Iii Ben F. Holt , Dietrich Robert A. , Parker Jane E. , Dangl Jeffery L.
Published: March 2003

LSD1 was defined as a negative regulator of plant cell death and basal disease resistance based on its null mutant phenotypes. We addressed the relationship between lsd1‐mediated runaway cell death and signaling components required for systemic acquired resistance (SAR), namely salicylic acid (SA) accumulation and NIM1/NPR1. We present two important findings. First, SA accumulation and NIM1/NPR1 are required for lsd1‐mediated runaway cell death following pathogen infection or application of chemicals that mimic SA action. This implies that lsd1‐dependent cell death occurs ‘downstream’ of the accumulation of SA. As SA application triggers runaway cell death in lsd1 but not wild‐type plants, we infer that LSD1 negatively regulates an SA‐dependent signal leading to cell death. Thus SA is both a trigger and a required mediator of lsd1 runaway cell death. Second, neither SA accumulation nor NIM1/NPR1 function is required for the basal resistance operating in lsd1. Therefore LSD1 negatively regulates a basal defense pathway that can act upstream or independently of both NIM1/NPR1 function and SA accumulation following avirulent or virulent pathogen challenge. Our data, together with results from other studies, point to the existence of an SA‐dependent ‘signal potentiation loop’ controlling HR. Continued escalation of signaling in the absence of LSD1 leads to runaway cell death. We propose that LSD1 is a key negative regulator of this signal potentiation.
Identification of peroxisomal targeting signal of pumpkin catalase and the binding analysis with PTS1 receptor
Authors: Kamigaki Akane , Mano Shoji , Terauchi Kaori , Nishi Yukiko , Tachibe‐Kinoshita Yuki , Nito Kazumasa , Kondo Maki , Hayashi Makoto , Nishimura Mikio , Esaka Muneharu
Published: March 2003

Many peroxisomal proteins are imported into peroxisomes via recognition of the peroxisomal targeting signal (PTS1) present at the C‐termini by the PTS1 receptor (Pex5p). Catalase, a peroxisomal protein, has PTS1‐like motifs around or at the C‐terminus. However, it remains unclear whether catalase is imported into peroxisome via the PTS1 system. In this work, we analyzed the PTS of pumpkin catalase (Cat1). A full or truncated pumpkin Cat1 cDNA fused at the 3′ end of the green fluorescent protein (GFP) coding sequence was introduced and stably expressed in tobacco BY‐2 (Nicotiana tabacum cv. Bright Yellow 2) cells or Arabidopsis thaliana by Agrobacterium‐mediated transformation. The cellular localization of GFP was analyzed by fluorescence microscopy. The results showed that the C‐terminal 10‐amino acid region containing an SKL motif‐like tripeptide (SHL) was not required for the import into peroxisomes. Surprisingly, the C‐terminal 3‐amino acid region was required for the import when the fusion proteins were transiently expressed by using particle gun bombardment, suggesting that the transient expression system is inadequate to analyze the targeting signal. We proposed that the C‐terminal amino acid region from 13 to 11 (QKL), which corresponds with the PTS1 consensus sequence, may function as an internal PTS1. Analysis of the binding of Cat1 to PTS1 receptor (Pex5p) by the yeast two‐hybrid system revealed that Cat1 can bind with the PTS1 receptor (Pex5p), indicating that Cat1 is imported into peroxisomes by the PTS1 system.
Dental management of children with latex allergy
Authors: Hashim Nainar S. M.
Published: March 2003

This paper reviews the aetiology, epidemiology and dental management of children with latex allergy. The issue of latex allergy has serious consequences for the dental management of children with one or more of the following risk factors: spina bifida, atopy, first surgery before one year of age, history of multiple surgical procedures, congenital urologic abnormalities, gastrointestinal malformations, hydrocephalus internus, ventriculo‐peritoneal shunts, spinal cord injuries, and family history of atopy. Management of latex allergy is based upon the diligent avoidance of latex exposure. Universal use of powder‐free low‐allergen latex gloves is recommended.
Ultrasound in the quantification of female pelvic organ prolapse
Authors: Dietz H. P. , Haylen B. T. , Broome J.
Published: March 2003

Objectives

To evaluate the use of ultrasound in the quantification of prolapse and compare findings with clinical assessments obtained in a blind study.

Methods

In a prospective comparative clinical study, 145 patients referred for urogynecological assessment were examined clinically by one operator and by translabial ultrasound by another operator.

Results

Clinical staging and International Continence Society coordinates were obtained for all 145 patients, as were ultrasound coordinates for descent of the anterior and posterior vaginal walls. Eighteen percent of the uteri of those women who had not had a hysterectomy in the past could not be seen; none of these women suffered from uterine prolapse clinically. Correlation with the prolapse assessment system recently endorsed by the International Continence Society was good (r = 0.77 for uterine prolapse, r = 0.72 for anterior vaginal wall and r = 0.53 for posterior vaginal wall descent).

Conclusions

This study demonstrates that translabial ultrasound can be used to quantify female pelvic organ prolapse. Correlation with the International Continence Society prolapse assessment system is good. The method may be particularly suitable for objective outcome assessment after surgical intervention. Copyright © 2001 International Society of Ultrasound in Obstetrics and Gynecology
Development of the human fetal corpus callosum: a high‐resolution, cross‐sectional sonographic study
Authors: Achiron R. , Achiron A.
Published: March 2003

Objective

To establish reference ranges during human pregnancy for normal fetal corpus callosum dimensions.

Design

In a prospective, cross‐sectional study of 258 fetuses between 16 and 37 weeks of gestation, measurements of the length, width, and thickness at the level of the anterior mid‐body of the corpus callosum were performed, using high‐resolution, transvaginal and transabdominal transducers.

Results

The mean length of the corpus callosum was 27.2 (standard deviation, 1.2; 95% confidence interval, 26.02–28.37) mm. Width and thickness of the corpus callosum were 5.6 (standard deviation, 1.6; 95% confidence interval, 5.41–5.82) mm and 1.9 (standard deviation, 0.7; 95% confidence interval, 1.87–2.06) mm, respectively. The size of the corpus callosum as a function of gestational age was expressed by regression equations: length (mm) = −20.40 + 1.92 × gestational age; width (mm) = −0.052 + 0.225 × gestational age; thickness (mm) = −0.174 + 0.085 × gestational age. The dimension–gestational age correlation coefficients were:r= 0.779 for length,r = 0.676 for width andr = 0.494 for thickness; these were statistically significant (P < 0.01). The maximum increase in thickness and width of the corpus callosum occurred between 19 and 21 weeks' gestation, while its length followed a constant growth rate. The normal mean length, width and thickness of the corpus callosum per week, and the 95% confidence limits, were defined.

Conclusions

The present study offers normative measurements of the fetal corpus callosum and may facilitate a more objective diagnosis of its congenital abnormalities. Copyright © 2001 International Society of Ultrasound in Obstetrics and Gynecology
Clinical implications of sonographic detection of uterine anomalies for reproductive outcome
Authors: Kupesic S.
Published: March 2003

The purpose of this review article was to analyze the literature on the prevalence, diagnosis and treatment of the congenital uterine anomalies and to discuss current dilemmas on their influence on reproductive outcome. Congenital uterine anomalies are commonly associated with repeated pregnancy failure, in particular an increased risk of first‐ and second‐trimester miscarriages and preterm delivery. Recent reports on two‐dimensional and three‐dimensional transvaginal ultrasound and saline contrast sonohysterography appear promising for diagnosis and classification of congenital uterine anomalies. The ability to visualize both the uterine cavity and the fundal uterine contour on a three‐dimensional scan facilitates the diagnosis of uterine anomalies and enables differentiation between septate and bicornuate uteri. Color Doppler ultrasound allows visualization of intraseptal vascularity and may help in distinguishing the avascular from the vascular septum. Less connective tissue in the septum may result in poor decidualization and placentation, while an increased amount of muscle tissue in the septum can cause miscarriage by the production of local uncoordinated myometrial contractility. Hysteroscopic septum excision is an effective procedure which improves live‐birth rates. Because of its simplicity, minimal invasiveness, low morbidity and low cost, a more liberal approach to the treatment of uterine anomalies is being advocated. Copyright © 2001 International Society of Ultrasound in Obstetrics and Gynecology
Ultrasonic evaluation of the uterus and uterine cavity after normal, vaginal delivery
Authors: Mulic‐Lutvica A. , Bekuretsion M. , Bakos O. , Axelsson O.
Published: March 2003

Objective

To describe uterine and uterine cavity changes throughout the puerperium, as revealed by ultrasound.

Methods

This was a prospective, longitudinal study in which 42 women with uncomplicated vaginal term deliveries were examined serially by ultrasound on postpartum days 1, 3, 7, 14, 28 and 56. The first four examinations were performed transabdominally and the last two transvaginally. The involution process of the uterus was assessed by measuring the anteroposterior diameter of the uterus and uterine cavity. Morphological findings were recorded. The influence on the involution process of parity, breast‐feeding, maternal smoking and infant's birth weight were also evaluated.

Results

The maximum anteroposterior diameter of the uterus diminished substantially and progressively from 92.0 mm on day 1 postpartum to 38.9 mm on day 56. The maximum anteroposterior diameter of the uterine cavity diminished from 15.8 mm on day 1 to 4.0 mm on day 56. However, the anteroposterior diameter of the uterine cavity, 5 cm from the fundus, typically increased on days 7 and 14 postpartum. The position of the uterus and the shape and the appearance of the cavity change in a unique way during the normal puerperium. The uterus was most often retroverted and empty in the early puerperium. Fluid and debris in the whole cavity were seen in the middle part of the puerperium. In late puerperium the cavity was empty and appeared as a thin white line. Endometrial gas was occasionally visualized. No correlation was found between the involution of the uterus and parity, breast‐feeding and the infant's birth weight.

Conclusion

Transabdominal sonography is suitable for examination of the uterus during the first 14 days postpartum but from day 28 the transvaginal route is preferable. The uterine body and position, as well as the cavity, are easy to examine by ultrasound. Accumulation of fluid and debris in the uterine cavity is a common and insignificant finding of the involuting uterus. It is located in the cervical area in the early puerperium and in the whole uterine cavity in the middle part of the puerperium. Findings from uncomplicated vaginal deliveries are needed as a reference when the diagnostic efficacy of ultrasound for pathological conditions is to be tested. Copyright © 2001 International Society of Ultrasound in Obstetrics and Gynecology
Effect of prophenoloxidase expression knockout on the melanization of microfilariae in the mosquito Armigeres subalbatus
Authors: Shiao S. H. , Higgs S. , Adelman Z. , Christensen B. M. , Liu S. H. , Chen C. C.
Published: March 2003

Melanization is an effective defence reaction used by mosquito hosts to kill malarial and filarial worm parasites. Although phenoloxidase (PO) has long been considered to be the key enzyme in the biosynthesis of melanotic material in insects, there is no direct evidence verifying its role in parasite melanization. To elucidate the role of PO in the melanization of microfilariae (mf) by mosquitoes, a double subgenomic Sindbis (dsSIN) recombinant virus was used to transduce Armigeres subalbatus mosquitoes with a 600 base antisense RNA targeted to the highly conserved copper‐binding region of an Ar. subalbatus PO gene. Compared with controls, haemolymph PO activity in mosquitoes transduced with antisense RNA was significantly reduced. When these mosquitoes were challenged with Dirofilaria immitis mf, the melanization of mf was almost completely inhibited. These data verify that PO is an essential component of the biochemical pathway required for the melanization of parasites, and that the dsSIN expression system represents a useful tool in the functional analysis of endogenous gene expression in mosquitoes.
Aedes aegypti transducing densovirus pathogenesis and expression in Aedes aegypti and Anopheles gambiae larvae
Authors: Ward T. W. , Jenkins M. S. , Afanasiev B. N. , Edwards M. , Duda B. A. , Suchman E. , Jacobs‐Lorena M. , Beaty B. J. , Carlson J. O.
Published: March 2003

Aedes aegypti densovirus (AeDNV) is a small DNA virus that has been developed into an expression and transducing vector for mosquitoes [Afanasiev et al. (1994) Exp Parasitol 79: 322–339; Afanasiev et al. (1999) Virology 257: 62–72; Carlson et al. (2000) Insect Transgenesis: Methods and Applications (Handler, A.M. & James, A.A., eds), pp. 139–159. CRC Press, Boca Raton]. Virions carrying a recombinant genome expressing the GFP gene were used to characterize the pathogenesis of the virus in 255 individual Aedes aegypti larvae. The anal papillae of the larvae were the primary site of infection confirming previous observations (Afanasiev et al., 1999; Allen‐Muira et al. (1999) Virology 257: 54–61). GFP expression was observed in most cases to spread from the anal papillae to cells of the fat body, and subsequently to many other tissues including muscle fibers and nerves. Infected anal papillae were also observed to shrink, or melanize and subsequently fall off in a virus dependent manner. Three to four day‐old larvae were less susceptible to viral infection and, if infected, were more likely to survive into adulthood, with 14% of them still expressing GFP as adults. Higher salt concentrations of 0.10–0.15 m inhibited viral infection. Anopheles gambiae larvae also showed infection of the anal papillae (17%) but subsequent viral dissemination did not occur. The persistence of the reporter gene expression into adulthood of Aedes aegypti indicates that transduction of mosquito larvae with recombinant AeDNV may be a means of introducing a gene of interest into a mosquito population for transient expression.
Differential expression of individual gene copies from within a tRNA multigene family in the mulberry silkworm Bombyx mori
Authors: Srinivasan L. , Gopinathan K. P.
Published: March 2003

In mulberry silkworm Bombyx mori, tRNA1Gly constitutes a multigene family from which the individual members are transcribed at different levels in vitro in homologous nuclear extracts. We report here the quantification of functional transcripts of these gene copies in vivo in B. mori‐derived BmN cells based on a suppression assay. The gene copies were converted to encode suppressor tRNAs and co‐transfected into cell lines with reporter gene(s) harbouring one or more nonsense mutations and the reporter gene activity was quantified. Individual members of the gene family were transcribed to very high‐, medium‐ and very low‐levels, following the same pattern as in vitro. All these gene copies were maximally expressed in Bm cells as compared to other insect cell lines.
Germline transformation of the malaria vector, Anopheles gambiae, with the piggyBac transposable element
Authors: Grossman G. L. , Rafferty C. S. , Clayton J. R. , Stevens T. K. , Mukabayire O. , Benedict M. Q.
Published: March 2003

Germline transformation of the major African malaria vector, Anopheles gambiae, was achieved using the piggyBac transposable element marked with the enhanced green fluorescent protein (EGFP) injected into mosquito embryos. Two G1 generation male mosquitoes expressing EGFP were identified among 34 143 larvae screened. Genomic Southern data and sequencing of the piggyBac insertion boundaries showed that these two males arose from one piggyBac insertion event in the injected G0 embryos. Genetic cross data suggest that the insertion site of the element either resulted in, or is tightly linked to, a recessive lethal. This was demonstrated by a deficiency in the number of EGFP‐expressing offspring from inbred crosses but expected ratios in outcrosses to non‐transformed individuals and failure to establish a pure‐breeding line. The insertion was weakly linked to the collarless locus on chromosome 2 and was shown by in situ hybridization to be located in division 28D of that chromosome. Particularly high levels of expression were observed uniformly in salivary glands and, in most individuals, in the anterior stomach. An improvement in the injection technique at the end of the studies resulted in increased G0 hatching, transient expression and EGFP‐expression rates among G1 progeny.
Evidence for genetic differentiation between the molecular forms M and S within the Forest chromosomal form of Anopheles gambiae in an area of sympatry
Authors: Wondji C. , Simard F. , Fontenille D.
Published: March 2003

We studied genetic variation at ten microsatellite DNA loci in Anopheles gambiae populations from the Forest chromosomal form collected in four villages in Cameroon (Central Africa). Both recently described M and S molecular forms occur in sympatry in this area. Geographic differentiation within form was low (Fst < 0.017) despite geographical distance between collection sites ranging from 35 to 350 km. However, higher (Fst > 0.035) and statistically significant levels of genetic differentiation were observed between forms, being the highest between sympatric M and S populations collected within the same village. Results were consistent across all loci spread throughout the genome, therefore reflecting a genome‐wide pattern. Considering previous findings of strong assortative mating within forms and general lack of hybrids in areas of sympatry, we propose that there is now sufficient direct and indirect evidence to consider both M and S molecular forms of An. gambiae as distinct species that have probably speciated recently.
Population structure of the tsetse fly Glossina pallidipes estimated by allozyme, microsatellite and mitochondrial gene diversities
Authors: Krafsur E. S.
Published: March 2003

Diversities at nuclear and mitochondrial loci were examined in eleven natural populations of Glossina pallidipes from east and southern Africa. Alleles in each class of loci are assumed to be selectively neutral. Allozyme gene diversities (heterozygosities) averaged over eight loci were 0.146 among seven Kenya populations and 0.201 among four southern African populations. Microsatellite diversity averaged over three loci was 0.250 in Kenya and only 0.218 in southern Africa. Mitochondrial diversities averaged 0.504 in Kenya and only 0.156 in southern Africa. Mitochondrial and microsatellite diversities in the populations were strongly correlated with each other, but uncorrelated with allozyme diversities. In contrast to the allozyme diversities, mitochondrial and microsatellite variation indicated a severe and prolonged reduction in population size in southern Africa. Genetic distances among populations increased with the geographical distances between them. Allozyme diversities in southern populations were conserved. Genetic differentiation at allozyme loci among populations was greatly damped when compared with the other markers. The foregoing can be explained if allozyme diversities were maintained by balancing selection. Three main points emerged: genetic data confirm the historical evidence that southern G. pallidipes populations experienced a severe and prolonged bottleneck; allozyme variation was conserved in the bottlenecked populations; and gene flow among populations is surprisingly restricted.
Intron‐dependent stimulation of marker gene expression in cultured insect cells
Authors: Zieler H. , Huynh C. Q.
Published: March 2003

We tested in a systematic fashion the effect of an intron on the level of luciferase expression in cultured C6/36 Aedes albopictus cells. The intron was inserted in both orientations, upstream and downstream of the luciferase coding region in two different luciferase expression vectors. The two parental luciferase expression vectors differed only in their promoters, one containing the Drosophila melanogaster actin5C promoter and the other the Autographa californica nuclear polyhedrosis virus hr5/ie1 enhancer/promoter. All resulting intron‐containing constructs were tested for their ability to express luciferase in transient assays following electroporation into C6/36 cells. We found that the introns stimulate luciferase expression between twelve and sixtyfold, depending on the promoter. Enhanced expression was only seen when the intron was present in the correct orientation upstream of the luciferase ORF. When the 3′ splice sites of the enhanced intron‐containing constructs were mutated, the expression level dropped back to below the level of the intronless parental constructs, suggesting that the intron‐dependent stimulation of luciferase expression is depending on splicing and is not due to other effects the intron may have on transcription or translation. The luciferase transcripts of all constructs were analysed by reverse transcription, PCR amplification and sequencing, and the results show a perfect correlation between efficient splicing of the intron and elevated levels of luciferase expression. Our findings have the potential to be very useful for boosting expression of foreign proteins in the widely used baculoviral or non‐viral systems in insect cells.
Phylogeography of the barbel (Barbus barbus) assessed by mitochondrial DNA variation
Authors: Kotlík P. , Berrebi P.
Published: March 2003

Using the phylogeographic framework, we assessed the DNA sequence variation at the mitochondrial cytochrome b gene across the distribution range of the barbel Barbus barbus, a widely distributed European cyprinid. Reciprocal monophyly of non‐Mediterranean European and Balkan/Anatolian populations is taken as evidence for a long‐term barrier to gene flow, and interpreted as a consequence of survival of the species in two separate refugia during several later glacial cycles. Lack of profound genealogical divergence across Europe from western France to the northwestern Black Sea basin is consistent with recent colonization of this area from a single glacial refuge, which was probably located in the Danube River basin. This may have occurred in two steps: into the Western European river basins during the last interglacial, and throughout the Central European river basins after the last glacial. The populations from the Balkans and Anatolia apparently did not contribute mitochondrial DNA to the post‐Pleistocene colonization of non‐Mediterranean Europe. Lack of detectable variation within the Balkans/Anatolia is attributed mainly to recent expansion throughout these regions, facilitated by the freshwater conditions and seashore regression in the Black Sea during the Late Pleistocene and Early Holocene.
Genetic diversity of indigenous Rhizobium leguminosarum bv. viciae isolates nodulating two different host plants during soil restoration with alfalfa
Authors: Zhang X.‐X. , Kosier B. , Priefer U. B.
Published: March 2003

A total of 360 Rhizobium leguminosarum bv. viciae strains was isolated from three brown‐coal mining restoration fields of different age and plant cover (without and in the first and second year of alfalfa, Medicago sativa, cultivation) using two host species (Viciahirsuta and Pisumsativum) as capture plants. The strains were genetically typed by restriction fragment length polymorphism analysis of polymerase chain reaction (PCR)‐generated 16S‐23S ribosomal DNA intergenic spacer regions (IGS–RFLP) and characterized by plasmid profiles and RFLP analysis of amplified nodABC genes. The R. leguminosarum bv. viciae population was dominated by the same group of strains (irrespective of the trap plant used). According to type richness, the genetic diversity of indigenous R. leguminosarum in the second year of restoration was lower than in the first year and it resembled that of the fallow field, except for plasmid types, in which it was higher than that of the fallow field. Some of the less frequent nodABC genotypes were associated with distinct chromosomal IGS genotypes and symbiotic plasmids (pSyms) of different sizes, indicating that horizontal transfer and rearrangements of pSym can occur in natural environments. However, the dominant pSym and chromosomal genotypes were strictly correlated suggesting a genetically stable persistence of the prevailing R. leguminosarum bv. viciae genotypes in the absence of its host plant.
Chloroplast DNA diversity in Calluna vulgaris (heather) populations in Europe
Authors: Rendell S , Ennos R. A
Published: March 2003

Variation in the chloroplast genome of Calluna vulgaris (heather), the dominant species of northwest European heath communities, was analysed using polymerase chain reaction–restriction fragment length polymorphisms (PCR–RFLPs) and microsatellites. No length polymorphisms were detected in the 100–200 base pair (bp) fragments amplified by the conserved microsatellite primers, and sequencing revealed that the repeat regions were interrupted relative to the corresponding sequence in Nicotiana tabacum. In contrast, PCR–RFLP analysis revealed high levels of haplotype diversity within populations (hS = 0.443, hT = 0.842), as well as substantial differentiation between populations (GST = 0.473). Diversity and differentiation were higher in southern Europe than in northern Europe. Interpreted in the light of data from allozyme studies and pollen core records, the results suggest that the main glacial refugia for C. vulgaris were located in southwest Europe, including northern Spain, the Pyrenees and the Massif Central region of France. There is also evidence for diffuse survival of the species at more northerly latitudes.
Phylogeographical structure and regional history of the dusky‐footed woodrat,Neotoma fuscipes
Authors: Matocq Marjorie D.
Published: March 2003

The dusky‐footed woodrat,Neotoma fuscipes, is a medium‐sized rodent that inhabits low elevation woodland habitats along the Pacific coast of North America from Oregon, throughout California and into Baja California. Analyses of mitochondrial sequence variation throughout the distribution reveal substantial phylogeographical structure withinN. fuscipes. The major mitochondrial lineages are largely concordant with previously identified morphological subdivisions within the taxon. The geographical distribution of distinct clades suggests that a combination of topographic barriers and the expansion and contraction of suitable habitat during the past 2 million years, especially along particular mountain ranges, have played a major role in the diversification ofN. fuscipes. Furthermore, relatively low levels of genetic variation across the northern half of the distribution suggest that dusky‐footed woodrats may have only recently expanded into this region.
Cross‐species microsatellite markers for elucidating population genetic structure in Arabidopsis and Arabis (Brassicaeae)
Authors: Clauss M. J. , Cobban H. , Mitchell‐Olds T.
Published: March 2003

Species closely related to model organisms present the opportunity to efficiently apply molecular and functional tools developed by a large research community to taxa with different ecological and evolutionary histories. We complied 42 microsatellite loci that amplify under common conditions in four closely related Arabidopsis: A. thaliana; A. halleri; A. lyrata ssp. lyrata; and A. lyrata ssp. petraea, as well as in one more distantly related crucifer; Arabis drummondii. Variation at these loci is amenable to a diversity of applications including population genetics, phylogeographical analyses, mapping of inter and intraspecific crosses, and recombination mapping. Our analysis of microsatellite variation illustrates significant differences in population genetic parameters among three Arabidopsis species. A population of A. thaliana, an inbreeding annual plant associated with disturbed habitats, was highly monomorphic (P = 8% percent polymorphic loci) and only 0.2% heterozygous for 648 locus‐by‐individual combinations. A population of the self‐incompatible perennial herb, A. halleri, was more genetically variable (P = 71%) and had an excess of heterozygosity that may reflect a recent population bottleneck associated with human‐mediated founder events. A population of the self‐incompatible perennial herb, A. lyrata ssp. petraea, was even more genetically variable (P = 86%) and appeared to be at mutation‐drift equilibrium. Population structure estimated from neutrally evolving loci provides an empirical expectation against which hypotheses of adaptive evolution at functional loci can be tested.
The spermatozoa and eggs of the cardinal fish
Authors: Lahnsteiner F.
Published: March 2003

The gametes of the cardinal fish Apogon imberbis had the following characteristic: the semen contained both biflagellate ( c. 20% in the investigated samples) and monoflagellate ( c. 80%) spermatozoa. The spermatozoa were acrosomless, had an ovoid head, and a cylindrical midpiece. The midpiece contained a high number of mitochondria (seven–10). The distal centriole was fastened to the nucleus via electron‐dense material. The proximal centriole was completely reduced. The eggs were spherical, had a smooth surface and one micropyle consisting of the micropylar channel, the vestibulum and a ring‐like convulsion bordering the vestibulum. The zona radiata was formed out of four layers which could be distinguished by their fine structural and histochemical features. It was only c. 1·5 µm thick and therefore much thinner than in other marine teleosts. The internal organization of the eggs (homogenous protein yolk containing cortical granules and lipid droplets) as well as the qualitative and quantitative biochemical composition of proteins, lipids and carbohydrates was similar to that of the pelagic eggs of other marine species.
Annual variation in gobiid larval density in the northern Baltic Sea
Authors: Parmanne R. , Lindström K.
Published: March 2003

The highest densities of gobiid larvae along the Finnish Baltic coast from 1974 to 1996 were found in the Åland Islands and the Archipelago Sea. The annual variation in gobiid densities was three‐fold (from 1· 1 to 3·3 larvae m−2) with peak densities in the years 1977 and 1992 and the lowest density in 1987. No long‐term trends in the density were found. The earliest gobiids were caught in the first half of May and the occurrence of larvae peaked at the end of June to early July.Within breeding seasons there was a positive correlation between water temperature at a depth of 1 m and the density of larvae. The density of larvae, however, did not correlate with mean summer temperature or the mean temperature of the previous summer. Instead larvae production was negatively affected by the severity of the previous winter as indicated by the maximum ice cover in the Baltic. The density of larvae was independent of larval density of the previous year suggesting that gobiid production primarily depends on the spawning stock size and may hence be regulated by density‐independent factors.
Nephrotic syndrome and mizoribine in children
Authors: Honda Masataka
Published: March 2003

AbstractBackground: The treatment of nephrotic syndrome (NS) has recently made dramatic progress. The ultimate purpose of treatment is that patients can lead a normal, disease‐free life with no adverse effects from treatment. However, clear treatment guidelines remain to be established in the children with frequent relapsing NS (FRNS) and steroid‐resistant NS (SRNS). The frequent use or large dose of steroids may lead to a serious adverse effect. Immunosuppressive drugs including cyclophasphamide and cyclosporin have been used to induce lasting remission, thereby sparing patients from further exposure to steroids. However, these drugs have both acute and chronic side‐effects. Mizoribine is a relatively safe and effective immunosuppressant. The report discusses the role of mizoribine in FRNS and SRNS in children.

Methods: The papers in experimental and clinical studies about mizoribine in NS were reviewed. Our experiences of mizoribine were also added.

Results: Mizoribine has been reported as an effective and safe drug for patients with FRNS. However, the efficacy of mizoribine has differed among various reports, depending on the dose given. There have yet to be any conclusive reports on the effects of mizoribine in SRNS in children.

Conclusions: Better results might be obtained if the doses of mizoribine increased in children with FRNS. When 5 mg/kg was used, no serious adverse effects were seen, therefore this dose may be safe and effective. An investigation of appropriate, effective and safe doses of mizoribine should be examined in the future. In patients with SRNS, large doses of mizoribine of more than 5 mg/kg might be effective, while the combined use of mizoribine and cyclosporin or methylpredonisolone pulse therapy might also be even more effective. Further studies are called for to examine the use of large doses of mizoribine.
Multiparametric flow cytometric analysis in a breast cancer cell line (MCF‐7)
Authors: Lee Gui Se Ra , Ryu Ki Sung , Rha Jong Gu , Kim Soo Pyung , Namkoong Sung Eun , Han Ku Taek
Published: March 2003

Objective:

To investigate whether multiparameter flow cytometric analysis of solid tumor specimens, including gynecologic tumors, which were stained triply with phycoerythrin (PE), fluorescein isothiocyanate (FITC) and propidium iodide (PI), can be performed simultaneously without interference from normal diploid cell populations and spectral overlap on a standard flow cytometer.

Methods:

MCF‐7 breast cancer cell lines and heterogeneous cell populations mixed with MCF‐7 cells and human peripheral blood lymphocytes (PBL) were fixed with 1% paraformaldehyde and permeabilized with 100% methanol. Cytokeratin and several proliferation‐associated cellular antigens (proliferating cell nuclear antigen (PCNA), p53, c‐erbB/2 and c‐myc) were labeled with PE and FITC, which was followed by DNA staining using PI. These labeled cells were measured on a standard FACScan flow cytometer equipped with a 488 nm single laser.

Results:

The coefficient of variation (CV) of the G0G1 peak of MCF‐7 cells was 4.3 and the cell cycle phase fractions of G0G1, S and G2M were 44.9, 45.9 and 9.2%, respectively. Fluorescein isothiocyanate, PE and PI fluorescences were detected without interference. The MCF‐7 cells expressed cytokeratin, PCNA, p53, c‐erbB/2 and c‐myc antigen. In the heterogeneous population of MCF‐7 cells mixed with PBL, two cellular populations were clearly separated into diploid PBL and aneuploid MCF‐7 cells without interference. The CV of G0G1 peak of PBL was 2.3 and the G0G1, S and G2M phase fractions were 85.5, 2.7 and 11.8%, respectively. The DNA index of MCF‐7 cells was 1.7, which indicated that the MCF‐7 cell line was composed of tumor cells with aneuploid DNA. The CV of the G0G1 peak of the MCF‐7 cells was 4.2, and the cell cycle phase fractions were 47.5% for G0G1, 42.3% for S, and 10.2% for G2M. The MCF‐7 cells expressed cytokeratin, but the PBL did not.

Conclusions:

Multiparameter flow cytometer analysis was useful to determine DNA ploidy status, phase fraction of the cell cycle and expression of cellular antigens and selective cytokeratin expression allowed epithelial originated tumor cells to be differentiated from normal stromal cells. This analysis could be performed without interference of spectral overlaps of fluorochromes using software‐based algorithmic compensation of spectral overlaps. Thus, this method offers new possibilities for multiparameter flow cytometric analysis and its use should be extended to future studies of the diagnosis, treatment and prediction of prognosis of the neoplasm.