bluuebunny/crossref_metadata_2025 huggingface dataset:
Quinacrine fluorescent karyotypes of human diploid and heteroploid cell lines
Authors: Miller O.J. , Miller D.A. , Allderdice P.W. , Dev V.G. , Grewal M.S.
Published: April 2008

Quinacrine-fluorescence karyotypes were prepared on a series of human cell lines. WI-38 karyotypes were indistinguishable from those obtained from cultured XX blood leukocytes. WI-L2 lymphoblastoid cells were 46, XY and pseudodiploid, with a single chromosome 21 and an additional submetacentric marker whose short arm may have been derived from a No. 21. Three HeLa cell lines had several marker chromosomes in common. Three of these marker chromosomes were also present in a D98/AG line, which therefore must have been of HeLa cell origin. Another cell line of presumptive human origin was shown to be a contaminant, of mouse origin.
Identification of the mouse chromosomes by quinacrine mustard staining
Authors: Francke Uta , Nesbitt Muriel
Published: April 2008

Metaphase spreads from male and female mice of various inbred and random-bred types were stained with the DNA-binding fluorochrome quinacrine mustard. This treatment caused the chromosomes to display cross-striation patterns of bright and dark bands visible with the fluorescence microscope. The intensity distribution of fluorescence along each chromosome was found to be distinctive and reproducible, so that it was possible to differentiate and identify all chromosome pairs in the mouse karyotype. On the average, the X chromosome was the fourth largest pair in female cells. The Y was usually the smallest chromosome and showed the strongest fluorescence. Densitometer tracing of fluorescent chromosomes resulted in distinctive density profiles which could assist in chromosome identification.
Assignment of linkage groups VIII and X to chromosomes in Mus musculus and identification of the centromeric end of linkage group I
Authors: Miller O.J. , Miller Dorothy A. , Kouri R.E. , Dev V.G. , Grewal M.S. , Hutton J.J.
Published: April 2008

The mitotic chromosomes in primary embryonic cultured cells of eight translocation stocks of Mus musculus were identified by their distinctive fluorescent banding patterns after staining with quinacrine mustard. By correlation of cytologic and genetic information, linkage group (LG) X was assigned to chromosome 7 and LG VIII to chromosome 5. Several earlier assignments were confirmed: LG I to chromosome 8, LG V to chromosome 2, LG XII to chromosome 19, LG XIII to chromosome 1, and LG XVIII to chromosome 9. In addition, chromosomes 4, 17, and 18 were identified in translocations, but the available genetic data did not permit linkage-group assignments. The centromere of chromosome 8 was located at the nv (Nijmegen waltzer) end of LG I by the two-breakpoint method. The position of the centromere of chromosome 1 at the fz (fuzzy) end of LG XIII and that of chromosome 9 at the nr (nervous) end of LG XVIII were confirmed by the same method.
Germ-cell chromosomes and their behavior during meiosis in a male Indian muntjac, Muntiacus muntjak
Authors: Sharma T.
Published: April 2008

Chromosome studies of testicular cells corroborated an earlier report of only seven chromosomes in somatic cells of males of the Indian muntjac, Muntiacus muntjak. The heterotrivalent observed during meiosis confirmed the view that a translocation had occurred between the X and an acrocentric autosome. Thus the male has an XYY, sex-chromosome system.
Diploid-polyploid cryptic species pairs: a possible clue to evolution by polyploidization in anuran amphibians
Authors: Bogart J.P. , Wasserman A.O.
Published: April 2008

Three of the seven currently recognized bisexual polyploid anuran species have morphologically very similar diploid “cryptic species” counterparts. The North American diploidtetraploid cryptic species pair Hyla chrysoscelis and H. versicolor is analyzed and compared with diploid and tetraploid populations of the South American nominal species Odontophrynus americanus. It is postulated that, through an intermediate triploid stage, tetraploidy could arise independently in diploid species by way of suppressed maturation in oöcytes. Tetraploid populations arising sympatrically from diploid species may maintain their integrity bio-acoustically. Polyploidization in the Anura is an important evolutionary mechanism and may prove to be quite wide spread when species are carefully examined on a population level.
Absence of correlation between XX/XY chimerism in the liver and the first signs of freemartinism in the calf fetus
Authors: Vigier B. , Prepin J. , Jost A.
Published: April 2008

XX/XY chimerism has been studied in the liver of 22 presumptive freemartin fetuses and in 12 male cotwins of multiple pregnancies, at an age of 39 to 62 days post-insemination. At these stages, the majority of the liver cells are hematopoietic cells. In these fetuses, the degree of chimerism varies from 0% to 55 %. Except for one case, the cells of the host predominate, i.e., the percentage of XY cells in freemartins is less than 50 % and is of the same order of magnitude as the number of XX cells in the male twin. According to several authors, freemartins and their male twins, when studied postnatally, possess the same percentage of XY (or XX) blood cells (variations from 1 % to almost 100%). These data suggest that the degree of chimerism of blood cells might change during development. If this is so, it seems unwarranted to compare chimerism in postnatal freemartins with the sex anomalies which appeared at an early fetal stage. The first signs of freemartinism (inhibition of gonadal growth and regression of the upper part of the Müllerian ducts) appear after day 48. In 15 freemartin fetuses of 49 to 62 days of age, the degree of inhibition of the left ovary and of the upper Müllerian ducts was compared with the percentage of XY cells present in the liver. The degree of inhibition of the ovarian volume and of the diameter of the upper Müllerian ducts was evaluated by reference to regression lines which had been previously calculated for a larger number of freemartins of the same age. There is no correlation between the percentage of XY cells and the degree of inhibition of the ovaries or of the Müllerian ducts.
A 45, XX,5–,13–, dic+ karyotype in a case of cri-du-chat syndrome
Authors: Niebuhr E.
Published: April 2008

A 45,XX,5–,13–, dic+ karyotype, verified by autoradiography and fluorescence studies, was found in a nine-year-old patient with typical clinical features of the cri-du-chat syndrome. A dicentric translocation chromosome was present in 34% of about 250 lymphocyte or fibroblast metaphases examined. Twisting in the intercentromeric region, leading to criss-cross separations, might explain the presence in 4% of the cells of a 46, XX,5p– karyotype. Other metaphases showed only one centromere on the translocation chromosome, which resembled a No. 1 in 49% of the cells and a No. 2 in 10%. Fluorescence studies indicated the presence of a centromere from a No. 5 and from a No. 13, respectively, on these monocentric chromosomes, and possible mechanisms leading to these structural alterations are discussed. Translocation chromosomes differing from these main types were seen in 3% of the metaphases. A fragile site on the long arm of one chromosome No. 16 was found in the father, a paternal aunt, and her son.
Sex-related mortality in developing chicken (Gallus domesticus) embryos
Authors: Keller C.A.
Published: April 2008

A total of 528 fertilized chick eggs (including 48 nonfertile eggs) were examined for viability and chromosomal sex after 60 to 90 h of incubation. Of those that contained embryos, 196 were male (ZZ), 183 were female (ZW), and 101 were of undetermined sex. The embryonal mortality rate was 18% for both the homogametic and the heterogametic sex and 19% for the undetermined sex groups. The results are consistent with the hypothesis that the sex ratio among live embryos does not deviate from 50 % male during the early incubation period and that recessive sex-linked lethal genes do not contribute significantly to a difference in mortality between the sexes in an ordinary breeding flock. It is suggested that sexing early chick embryos by chromosome morphology may require relatively small samples to detect lethal recessives induced by potentially mutagenic agents.
Sex-chromosome translocations and karyotypes in bovid tribes
Authors: Wurster Doris H.
Published: April 2008

Chromosomes of species from four bovid tribes are discussed in this paper. Two of the tribes are characterized by species with translocations or other unusual features involving the sex chromosomes. The other two tribes are characterized by a complete lack of unusual sex-chromosome arrangements. Groupings by sex-chromosome characteristics are consistent with classical taxonomic categories.
Fluorescence of X and Y chromatin in human interphase cells
Authors: Mukherjee A.B. , Moser Gertrude C. , Nitowsky H.M.
Published: April 2008

The fluorescent characteristics of X (sex) and Y chromatin of human interphase nuclei stained with quinacrine mustard (QM) were studied in fibroblast-like cell cultures derived from several different tissues. X chromatin in female cells was clearly distinguishable from Y chromatin in male cells on the basis of size and fluorescent intensity. The frequency of X chromatin was dependent on cell density in vitro, whereas the frequency of Y chromatin was independent of cell density. The fluorescent intensity of X chromatin in cultured cells was much stronger than that in uncultured cells, such as those of the oral mucosa. The intensity of Y-chromatin fluorescence, however, was nearly the same in both cultured and uncultured cells. Nuclei first stained with QM, photographed, and then restained with thionin and rephotographed showed identical X- and Y-chromatin morphology.
Karyotype analysis of Xenopus muelleri (Peters) and Xenopus laevis(Daudin), Pipidae
Authors: Tymowska Janina , Kobel H.R.
Published: April 2008

A cytogenetic analysis of two anuran species, Xenopus laevis laevis and Xenopus muelleri, was performed. The diploid number of chromosomes in both species was 2n = 36. The two karyotypes, arranged according to size of chromosome and centromere position, showed no pronounced differences. However, the chromosomes which had a secondary constriction were different. In X. laevis this constriction (nucleolar organizer) was found on one pair of acrocentric chromosomes only, the pair showing association in about 50% of the metaphases. In X. muelleri, on the other hand, two distinct pairs of submetacentric chromosomes contained secondary constrictions. Neither of these two chromosome pairs was morphologically similar to the constriction-bearing chromosomes of X. laevis. Association similar to that in X. laevis was found in one pair only, the one with a terminal constriction. In neither of the two species was there any evidence of sex-chromosome heteromorphism.
The karyotype of the pig (Sus scrofa domestica), identified by quinacrine mustard staining and fluorescence microscopy
Authors: Hansen K.M.
Published: April 2008

The karyotype of the pig (Sus scrofa domestica) was characterized by means of quinacrine mustard staining and fluorescence microscopy. The material examined consisted of 15 pigs of the Danish Landrace, five boars and 10 sows, representing five boar lines and eight sow strains. Previously it had been possible to identify only six pairs of chromosomes and the Y chromosome among the 38 chromosomes of the pig. With the present method it was possible to identify both autosomes and sex chromosomes with great reliability. Fluorescence variants were not demonstrable.
The C-band and G-band patterns of Microtus agrestis chromosomes
Authors: Cooper J.E.K. , Hsu T.C.
Published: April 2008

The C-band and G-band patterns of Microtus agrestis metaphase chromosomes are described. The C-band pattern reveals constitutive heterochromatin as uniformly intenselystained areas but cannot aid in identifying autosomal pairs. The G-bands revealed by a heat renaturation method (ASG) were compared with those revealed by treatment with three proteolytic enzymes. All procedures yield apparently the same specific pattern of crossbands on the autosomes, and each autosomal pair can therefore be identified by its characteristic pattern. The constitutive heterochromatin of the sex chromosomes stains uniformly with the heat renaturation method but is subdivided into regions with different staining intensities with each enzyme treatment. A G-band karyotype for Microtus agrestis metaphase chromosomes is presented.
Trisomy for the smallest autosome of the mouse and identification of the T1Wh translocation chromosome
Authors: White Beverly J. , Tjio J.-H. , Van de Water Lisa C. , Crandall Clare
Published: April 2008

Quinacrine-mustard staining and Giemsa-banding identified the large chromosome of the T1Wh translocations as No. 5; the small autosome of the translocation had previously been found to be No. 19. Others have demonstrated that translocation T163H involved autosomes 9 and 19; these findings were confirmed in our study. Meiotic studies of (T1Wh × T163H) F1 hybrids showed a chain quadrivalent at metaphase I, confirming that both translocations shared in common only chromosome 19. Metaphase II analysis of F1 hybrids demonstrated frequent nondisjunction of the T1Wh and T163H translocation chromosomes, and 12 % of 75 F2 offspring from 16 F1 crosses were trisomic for chromosome 19. Trisomics had three translocation chromosomes and a chromosome number of 38. All newborn trisomics were smaller than their normal littermates and died during the first day of life. The only specific malformation found so far has been cleft palate.
Quantitative karyotyping: a model twin study
Authors: Neurath P.W. , Lin P.-S. , Low D.A.
Published: April 2008

Chromosome measurements of length and area, performed with the help of PIQUANT, our computer system, have been combined with the quinacrine-mustard fluorescent-banding technique to characterize the karyotype of individuals. Five sets of monozygotic twins were examined, and it was found that (1) individuals differ significantly in their chromosome measurements, particularly in the size, and (2) the within-twin pair differences are very small and not significant, thus establishing the genetic basis of the differences under (1). The observed differences are not only statistically significant but large enough to permit selection of all five sets of twins from only the chromosome measurements of their autosomes. The potential of these measurements to serve as genetic markers is shown to be considerable, whenever 10 to 30 cells per person, all banded, can be measured.
Metaphase cell finding by machine
Authors: Green D.K. , Cameron J.
Published: April 2008

The basic elements of a mitotic cell-finding machine are described. The hardware components comprising the machine are a motor-controlled microscope, a vidicon scanner and associated video-signal processor, and a digital computer. By very rudimentary signal processing, it is possible to derive an analog voltage from the camera video which is proportional to the amount of “speckled image” characteristic of metaphase cells at low magnification. A digital computer is used to control the microscope movement and to digitize the signal-processor analog output. Microscope slides prepared from human blood cultures are used in all the experiments. The results of a pilot experiment designed to measure the “speckled image” voltage for image features within a 100 µ by 100 µ field of view are discussed and extrapolated to the expected machine performance at larger and necessarily more complicated fields of view. It is expected that when conditions are set such that no usable metaphase cell image is missed by the machine, 50 % of the images scored will be usable, where a usable cell is one from which a karyotype could be obtained. Secondary picture processes for final selection of usable cells at high magnification are postulated, and it is estimated that the first- and second-stage searching processes together will give rise to an average cell finding rate of approximately 9 sec/cell. An experimentally tested automatic-focussing technique using the vidicon camera system is discussed.
Trisomy 19 in the laboratory mouse
Authors: White B.J. , Tjio J.-H. , Van de Water L.C. , Crandall C.
Published: April 2008

Weights of viable trisomic mouse fetuses and placentas were compared with those of normal littermates between 12½ and 19½ days’ gestation. Mean trisomic fetal and placental weights were below normal weights at all stages. Differences in mean body weight were consistently significant, whereas those in placental weight varied. Trisomic fetuses of a specific karyotype predisposing to cleft palate were not smaller than trisomics of two other karyotypes, suggesting that a greater generalized growth lag was not responsible for the cleft palate. At each developmental stage, ratios of placental to body weights of normal and trisomic mice were similar. Sections of trisomic organs showed only one specific change: excessive degeneration of oocytes in some trisomic ovaries. All of these findings are discussed in relation to other studies of fetal and placental weight in mice and to studies of the effects of trisomy upon human development in utero.
Translocation of centromeric heterochromatin in the T(10;13) 199H stock of Mus musculus and localization of chromosome break points
Authors: Dev V.G. , Miller D.A. , Charen J. , Miller O.J.
Published: April 2008

Combined quinacrine fluorescent (Q-banding) and centromeric heterochromatin (C-banding) staining of metaphase chromosomes from the T199H heterozygous translocation stock of Mus musculus has shown that the break points in chromosomes 10 and 13 are located closer to each centromere than previously believed. Virtually all of chromosome 13, including the C-banding material, has been translocated to the distal region of chromosome 10. The presence of C-banding material in an interstitial location in the longer of the two translocation chromosomes provides a distinctive mitotic chromosome marker. Both the C-banding region and the euchromatic region of chromosome 13 are longer in the translocation chromosome than in the normal homolog. During meiosis in heterozygous males a chain of four chromosomes is the most common quadrivalent configuration. In the meiotic chromosome the amount of interstitial C-banding is markedly reduced from that seen in mitosis. C-banding is seen at the centromeric region of both the short translocation chromosome and the Y during meiosis but not mitosis.
DNA helical content during the C-banding procedure
Authors: Kurnit D.M.
Published: April 2008

DNA was isolated from chromosomes or nuclei scraped off slides after each step of the C-banding procedure. Analysis by buoyant density-gradient centrifuga-tion and digestion with nucleases specific for single-stranded nucleic acids showed: 80 % or more of the DNA remained double-stranded after fixation to slides and 70 % or more was single-stranded after the slides were treated with alkali, whether the slides were annealed afterwards or not. Furthermore, highly repeated DNAs remained single-stranded after annealing following denaturation. Since the annealing of slides after denaturation does not effect the reassociation of either bulk or highly repeated DNA sequences, C-banding apparently results from DNA-protein interactions rather than preferential strand reassociation of highly repeated DNAs.
Chromosome homology in birds: banding patterns of the chromosomes of the domestic chicken, ring-necked dove, and domestic pigeon
Authors: Stock A.D. , Arrighi F.E. , Stefos K.
Published: April 2008

Improved techniques for culturing avian tissue in combination with a trypsinurea chromosome banding technique has allowed an analysis of chromosome homology in three bird species. Three macrochromosomes of the domestic chicken (Galliformes) showed homology with those of the ring-necked dove and the domestic pigeon (Columbiformes). The ring-necked dove and domestic pigeon displayed similar banding patterns, except that the pigeon possessed an additional four pairs of microchromosomes, which are fused to form two pairs of macrochromosomes in the dove. Both G- and C-banded metaphase chromosomes of the three species are presented, and chromosome evolution and phyletic relations are discussed.
Male heterogamety in kinosternid turtles (genus Staurotypus)
Authors: Bull J.J. , Moon R.G. , Legler J.M.
Published: April 2008

Male heterogamety, interpreted as an XX/XY sex-chromosome system, occurs in the turtle genus Staurotypus (family Kinosternidae). Meiotic analysis showed that the basis for the heteromorphism resides in relatively small regions of a pair of macrochromosomes. Heteromorphism was not observed in the three other genera belonging to the same family. Available data for reptiles and higher vertebrates strongly suggest that sex-chromosome heterogamety has evolved independently at least three times in reptiles (in turtles, lizards, and snakes) and, therefore, that the sex chromosomes of birds do not share a common ancestry with those of snakes.
Characteristics of an HPRT-deficient Chinese hamster cell line
Authors: Gee P.A. , Ray M. , Mohandas T. , Douglas G.R. , Palser H.R. , Richardson B.J. , Hamerton J.L.
Published: April 2008

A mutant Chinese hamster cell line was selected from wild-type CHW cells in 30 µg/ml 8-azaguanine, after exposure for 2 h to 10–3 M methyl methanesulphonate. This line was subsequently cloned and one clone, isolated in 10 µg/ml 8-azaguanine (CHW-1102), was selected for further study. CHW-1102 showed stability of resistance to 8-azaguanine after six months of growth in nonselective medium and an LD50 to 8-azaguanine of between 10 and 20 µg/ml, compared to 2.5 µg/ml for the wild-type CHW cell line. The doubling time in culture was found to be 13 h, compared to a doubling time of 12 h for CHW. Biochemical studies showed that the mutant cell line CHW-1102 had a specific activity of HPRT of about 1–2 nmol/mg protein per hour compared to a specific activity of the wild type of 200 nmol/mg protein per hour. The spontaneous reversion rate from 8-azaguanine resistance to 8-azaguanine sensitivity was 4.23 × 10–8/locus per generation. Both the parental line (CHW) and the mutant (CHW-1102) have a modal chromosome number of 22 and a relatively stable karyotype. Both lines show some chromosome rearrangement and, in particular, the presence of one long acrocentric marker chromosome, which consists largely of material from chromosomes 6 and 8. Much of the distal part of the long arm of the X chromosome, which is heterochromatic, has been lost in both CHW and CHW-1102. So far as can be determined, no autosomal material, except perhaps the paracentromeric region of chromosome 6, is missing in CHW-1102, although even this material may be represented by a small marker chromosome M2. Both cell lines show characteristics that are of value for the study of mammalian cell genetics, and CHW-1102 represents a useful addition to the stock of mutant mammalian cell lines.
Familial X/X translocation:t(X;X)(p22;q13)
Authors: Kim H.J. , Hsu L.Y.F. , Hirschhorn K.
Published: April 2008

We have studied a family with an X/X translocation which was found in a 16-year-old daughter and in her 34-year-old mother. The daughter had short stature, secondary amenorrhea, and many Turner’s stigmata. The mother, 5-ft tall, had a few Turner’s stigmata and had developed secondary amenorrhea at age 24. Both peripheral leukocyte and skin fibroblast cultures of the daughter revealed, by conventional karyotype analysis, 46 chromosomes with one C-group chromosome missing and with an extra No. 2-like chromosome in all metaphase cells examined. By both Q- and G-banding techniques, this abnormal karyotype was identified as 46,X, + t(X;X)(p22;ql3). An identical translocation chromosome was demonstrated in the mother’s cells, but 50 % of the cells had a 45,X chromosomal constitution. Autoradiographic studies showed that the X/X translocation chromosome was the late-replicating X in all cells examined from both the daughter and the mother. The difference in the phenotypic manifestations of the daughter and of her mother can be explained perhaps by the different contents of the other active X chromosomes. When the same X chromosome is inactivated in all cells, mutations present in the active X chromosome are not compensated and therefore are expressed.
The gorilla karyotype: chromosome lengths and polymorphisms
Authors: Miller D.A. , Firschein I.L , Dev V.G. , Tantravahi R. , Miller O.J.
Published: April 2008

Metaphase chromosome preparations of three male and one female Gorilla gorilla were stained to demonstrate quinacrine, Giemsa, centromeric heterochromatin, and, in one case, reverse-Giemsa bands. A standard karyotype is proposed based on chromosome banding pattern, centromeric index, and length. Three types of variation between homologous chromosomes are described: presence or absence of very bright fluorescence of the short arm or satellite region of acrocentric chromosomes, presence or absence of bright quinacrine bands at the distal ends of chromosome arms, and large differences in the size of the heterochromatic region on each of two biarmed chromosomes. At least half the chromosome pairs show polymorphisms of these types. Satellite associations were scored for each animal. In one case the two smallest pairs of chromosomes were preferentially involved in associations.
Synteny of the human loci for fumarate hydratase and UDPG pyrophosphorylase with chromosome 1 markers in somatic cell hybrids
Authors: van Someren H. , Beijersbergen van Henegouwen H. , Westerveld A. , Bootsma D.
Published: April 2008

The segregation of the loci for 26 human enzyme markers was studied in man-Chinese hamster somatic cell hybrids. The results showed synteny of the human loci for fumarate hydratase, UDPG pyrophosphorylase, phosphogluconate dehydro-genase, phosphoglucomutasei, and peptidase-C (chromosome 1). Furthermore, the synteny of the human loci for mannose phosphate isomerase and the leukocytic form of pyruvate kinase was confirmed.
Socio-Economic Inequality and Cultural Fragmentation in Western Societies
Authors: Janmaat Jan Germen
Published: April 2008

This article examines the relation between socio-economic inequality and disparities of democratic values in Western societies. It discusses three perspectives on democratic attitudes and values – rising inequality, social capital, and postmaterialism – and explores to what extent cross-national patterns and trends in value disparities are in agreement with the predicted outcomes of these perspectives. Use is made of the World Value Survey and the European Value Study to explore these value disparities. The results do not provide unequivocal support for any of the three perspectives. The patterns on some values are in line with the rising inequality perspective, while those on others are consistent with the other two perspectives. Low and high incomes have come to drift apart on democratic values, which is what the rising inequalities perspective would expect. But these widening disparities are unrelated to socio-economic inequalities. It is proposed that socio-economic inequalities primarily affect mean levels of democratic values while individualism is the key factor producing value divergence.
Biocompatible alumina ceramic for total hip replacements
Authors: Zeng P.
Published: April 2008

Their resistance to wear and biocompatibility make ceramics ideal materials for medical applications, such as implants. For over 30 years, pure alumina has been the dominant material for ceramic hip prostheses. Interest in alumina hip prostheses continues to grow, due to the relatively short life of polymer/metal prostheses, mainly resulting from osteolysis and aseptic loosening caused by polymer wear debris. Since its introduction by Boutin in the 1970s, substantial improvements have been achieved in the microstructure of medical grade alumina by improving purity and processing to give complete densification and fine, uniform grain sizes. A brief review is given of the types of alumina used in total hip replacement, the development of medical grade alumina, and methods of in vivo and in vitro investigation of alumina prostheses, with a focus on current knowledge of the damage observed on alumina prostheses. Particular attention is paid to wear mechanisms and the influence of materials properties on wear behaviour. A region of relatively severe wear, known as stripe wear, is widely observed on retrieved alumina hip prostheses. This type of wear can now be replicated in vitro in joint simulators by the introduction of a ‘microseparation’ motion during the test cycle. Finally, the future of ceramic hip prostheses and development of the next generation of ceramics for hip prostheses is discussed.
Mesenchymal stem cells exert differential effects on alloantigen and virus-specific T-cell responses
Authors: Karlsson Helen , Samarasinghe Sujith , Ball Lynne M. , Sundberg Berit , Lankester Arjan C. , Dazzi Francesco , Uzunel Mehmet , Rao Kanchan , Veys Paul , Le Blanc Katarina , Ringdén Olle , Amrolia Persis J.
Published: April 2008

Mesenchymal stem cells (MSCs) suppress alloantigen-induced T-cell functions in vitro and infusion of third-party MSCs seems to be a promising therapy for graft-versus-host disease (GVHD). Little is known about the specificity of immunosuppression by MSCs, in particular the effect on immunity to pathogens. We have studied how MSCs affect T-cell responses specific to Epstein-Barr virus (EBV) and cytomegalovirus (CMV). We found that EBV- and CMV-induced proliferation and interferon-γ (IFN-γ) production from peripheral blood mononuclear cells (PBMCs) was less affected by third-party MSCs than the response to alloantigen and that MSCs had no effect on expansion of EBV and CMV pentamer-specific T cells. Established EBV-specific cytotoxic T cells (CTL) or CMV-CTL cultured with MSCs retained the ability to proliferate and produce IFN-γ in response to their cognate antigen and to kill virally infected targets. Finally, PBMCs from 2 patients who received MSCs for acute GVHD showed persistence of CMV-specific T cells and retained IFN-γ response to CMV after MSC infusion. In summary, MSCs have little effect on T-cell responses to EBV and CMV, which contrasts to their strong immunosuppressive effects on alloreactive T cells. These data have major implications for immunotherapy of GVHD with MSCs and suggest that the effector functions of virus-specific T cells may be retained after MSC infusion.
Molecular and functional characterization of a Rho GDP dissociation inhibitor in the filamentous fungus Tuber borchii
Authors: Menotta Michele , Amicucci Antonella , Basili Giorgio , Polidori Emanuela , Stocchi Vilberto , Rivero Francisco
Published: April 2008

Background

Small GTPases of the Rho family function as tightly regulated molecular switches that govern important cellular functions in eukaryotes. Several families of regulatory proteins control their activation cycle and subcellular localization. Members of the guanine nucleotide dissociation inhibitor (GDI) family sequester Rho GTPases from the plasma membrane and keep them in an inactive form.

Results

We report on the characterization the RhoGDI homolog of Tuber borchii Vittad., an ascomycetous ectomycorrhizal fungus. The Tbgdi gene is present in two copies in the T. borchii genome. The predicted amino acid sequence shows high similarity to other known RhoGDIs. Real time PCR analyses revealed an increased expression of Tbgdi during the phase preparative to the symbiosis instauration, in particular after stimulation with root exudates extracts, that correlates with expression of Tbcdc42. In a translocation assay TbRhoGDI was able to solubilize TbCdc42 from membranes. Surprisingly, TbRhoGDI appeared not to interact with S. cerevisiae Cdc42, precluding the use of yeast as a surrogate model for functional studies. To study the role of TbRhoGDI we performed complementation experiments using a RhoGDI null strain of Dictyostelium discoideum, a model organism where the roles of Rho signaling pathways are well established. For comparison, complementation with mammalian RhoGDI1 and LyGDI was also studied in the null strain. Although interacting with Rac1 isoforms, TbRhoGDI was not able to revert the defects of the D. discoideum RhoGDI null strain, but displayed an additional negative effect on the cAMP-stimulated actin polymerization response.

Conclusion

T. borchii expresses a functional RhoGDI homolog that appears as an important modulator of cytoskeleton reorganization during polarized apical growth that antecedes symbiosis instauration. The specificity of TbRhoGDI actions was underscored by its inability to elicit a growth defect in S. cerevisiae or to compensate the loss of a D. discoideum RhoGDI. Knowledge of the cell signaling at the basis of cytoskeleton reorganization of ectomycorrhizal fungi is essential for improvements in the production of mycorrhized plant seedlings used in timberland extension programs and fruit body production.
Acute-phase reactants after paediatric cardiac arrest. Procalcitonin as marker of immediate outcome
Authors: Los Arcos Marta , Rey Corsino , Concha Andrés , Medina Alberto , Prieto Belen
Published: April 2008

Objective

Procalcitonin (PCT) and C reactive protein (CRP) have been used as infection parameters. PCT increase correlates with the infection's severity, course, and mortality. Post-cardiocirculatory arrest syndrome may be related to an early systemic inflammatory response, and may possibly be associated with an endotoxin tolerance. Our objective was to report the time profile of PCT and CRP levels after paediatric cardiac arrest and to assess if they could be use as markers of immediate survival.

Materials and methods

A retrospective observational study set in an eight-bed PICU of a university hospital was performed during a period of two years. Eleven children younger than 14 years were admitted in the PICU after a cardiac arrest. PCT and CRP plasma concentrations were measured within the first 12 and 24 hours of admission.

Results

In survivors, PCT values increased 12 hours after cardiac arrest without further increase between 12 and 24 hours. In non survivors, PCT values increased 12 hours after cardiac arrest with further increase between 12 and 24 hours. Median PCT values (range) at 24 hours after cardiac arrest were 22.7 ng/mL (0.2 – 41.0) in survivors vs. 205.5 ng/mL (116.6 – 600.0) in non survivors (p < 0.05). CRP levels were elevated in all patients, survivors and non-survivors, at 12 and 24 hours without differences between both groups.

Conclusion

Measurement of PCT during the first 24 hours after paediatric cardiac arrest could serve as marker of mortality.
Predictive factors for beneficial application of high-frequency electromagnetics for tumour vaporization and coagulation in neurosurgery
Authors: Ritz Rainer , Heckl Stefan , Safavi-Abbasi Sam , Feigl Guenther C , Krischek Boris , Lüdemann Wolf , Mirzayan Javed M , Koerbel Andrei , Samii Madjid , Tatagiba Marcos , Gharabaghi Alireza
Published: April 2008

Objective

To identify preoperative and intraoperative factors and conditions that predicts the beneficial application of a high-frequency electromagnetic field (EMF) system for tumor vaporization and coagulation.

Methods

One hundred three subsequent patients with brain tumors were microsurgically treated using the EMF system in addition to the standard neurosurgical instrumentarium. A multivariate analysis was performed regarding the usefulness (ineffective/useful/very helpful/essential) of the new technology for tumor vaporization and coagulation, with respect to tumor histology and location, tissue consistency and texture, patients' age and sex.

Results

The EMF system could be used effectively during tumor surgery in 83 cases with an essential contribution to the overall success in 14 cases. In the advanced category of effectiveness (very helpful/essential), there was a significant difference between hard and soft tissue consistency (50 of 66 cases vs. 3 of 37 cases). The coagulation function worked well (very helpful/essential) for surface (73 of 103 cases) and spot (46 of 103 cases) coagulation when vessels with a diameter of less than one millimeter were involved. The light-weight bayonet hand piece and long malleable electrodes made the system especially suited for the resection of deep-seated lesions (34 of 52 cases) compared to superficial tumors (19 of 50 cases).

The EMF system was less effective than traditional electrosurgical devices in reducing soft glial tumors. Standard methods where also required for coagulation of larger vessels.

Conclusion

It is possible to identify factors and conditions that predict a beneficial application of high-frequency electromagnetics for tumor vaporization and coagulation. This allows focusing the use of this technology on selective indications.
Assessing midwives' breastfeeding knowledge: Properties of the Newborn Feeding Ability questionnaire and Breastfeeding Initiation Practices scale
Authors: Creedy Debra K , Cantrill Ruth M , Cooke Marie
Published: April 2008

Background

There are few reliable and valid tools to assess lactation and infant feeding knowledge and practices. This study tested the psychometric properties of two new scales, the Newborn Feeding Ability (NFA) questionnaire and Breastfeeding Initiation Practices (BIP) scale to assess midwives' breastfeeding knowledge and practices specific to breastfeeding initiation.

Methods

A national postal survey of Australian midwives (n = 3500) was conducted in October 2001. Reliability was determined through Cronbach's alpha coefficient and stability determined by a test-retest. Content validity was established through a critical review of literature and review by an expert panel. Construct validity was informed by an exploratory factor analysis and principle component analysis with varimax rotation. Correlations between NFA and BKQ knowledge subscale scores and BIP and BKQ practice subscale scores assessed criterion validity. A multiple hierarchical regression analysis determined predictive validity of the NFA and BIP.

Results

A response rate of 31.6% (n = 1107) was achieved. Adequate internal consistency was established for both instruments. Five factors on the NFA questionnaire were congruent with knowledge about effects of skin-to-skin contact, physiological stability, newborn innate abilities, work practices and effective breastfeeding. The BIP revealed three factors related to observing pre-feeding behavior, mother/baby care and attachment and positioning practices. Predictive validity of knowledge was moderate (r = 0.481, p < 0.01) and contributed to 31.5% of variance in reported practice. Midwives with high knowledge scores were more likely to report best practice when assisting mothers to initiate breastfeeding. Midwives with more personal breastfeeding experience scored higher on all scales.

Conclusion

The Newborn Feeding Ability questionnaire and the Breastfeeding Initiation Practices scale can contribute to practice development by assessing lactation and infant feeding knowledge and practice deficits. Individual learning needs can be identified, and effectiveness of education interventions evaluated using these tools. Further testing is required with other samples of midwives and health professionals involved in the promotion of breastfeeding.
Splenic rupture as the presenting manifestation of primary splenic angiosarcoma in a teenage woman: a case report
Authors: Manouras Andreas , Giannopoulos Panagiotis , Toufektzian Levon , Markogiannakis Haridimos , Lagoudianakis Emmanuel E , Papadima Artemisia , Papanikolaou Dimitrios , Filis Konstantinos , Kekis Panagiotis
Published: April 2008

Introduction

Primary splenic angiosarcoma is a rare neoplasm of vascular origin carrying a very poor prognosis, partly due to its high metastatic potential. This disease presents frequently with splenic rupture and hemorrhage. We report the case of a 17-year-old woman who presented with rupture of a primary splenic angiosarcoma.

Case presentation

The patient presented with diffuse abdominal pain and distention. Clinical examination revealed severe tenderness in the left upper abdominal quadrant, a palpable abdominal mass, and hemodynamic instability with a systolic arterial blood pressure of 75 mmHg and heart rate of 135 beats per minute. Blood tests revealed anemia (hemoglobin 7.0 g/dl) and thrombocytopenia (platelets 70 × 109/liter). After initial fluid resuscitation and stabilization, abdominal ultrasound and computed tomography were performed, revealing a large quantity of intraperitoneal free fluid, an enlarged spleen, and a heterogeneous low-density signal within the splenic parenchyma, which showed varying degrees of contrast enhancement. At laparotomy a huge (weight 1530 g, diameter 19 cm) actively bleeding spleen was identified and splenectomy was performed. Histopathology showed a primary splenic angiosarcoma. After an uneventful recovery, the patient was discharged on the sixth postoperative day.

Conclusion

Primary splenic angiosarcoma is rare. Although this malignancy is usually encountered in advanced age, there have been a few reported cases among younger patients. The case reported here presented with splenic rupture, was treated by laparotomy and splenectomy, and the patient is disease free 16 months after surgery.
Morphologically and immunohistochemically undifferentiated gastric neoplasia in a patient with multiple metastatic malignant melanomas: a case report
Authors: Alghisi Federico , Crispino Pietro , Cocco Andrea , Richetta Antonio G , Nardi Francesco , Paoluzi Paolo , Badiali Danilo
Published: April 2008

Introduction

Malignant melanoma is a neoplasia which frequently involves the gastrointestinal tract (GIT). GIT metastases are difficult to diagnose because they often recur many years after treatment of the primary cutaneous lesion and also manifest clinically at an advanced stage of the neoplasia. Furthermore, GIT metastases can appear in various morphological forms, and therefore immunohistochemistry is often useful in distinguishing between a malignant melanoma and other malignancies.

Case presentation

We report the case of a 60-year-old man with a multiple metastatic melanoma who underwent an upper endoscopy to clarify the possible involvement of the gastric wall with a mass localized in the upper abdomen involving the pancreas and various lymph nodes, which was previously described with computed tomography. Clinically, the patient reported a progressive loss of appetite, nausea and vomiting. The upper endoscopy and histological examination revealed a gastric location of an undifferentiated neoplasm with an absence of immunohistochemical characteristics referable to the skin malignant melanoma that was removed previously.

Conclusion

The present case report shows the difficulty in diagnosing a metastatic melanoma in the GIT and therefore, it seems worthwhile to consider metastatic malignant melanoma in the differential diagnosis of undifferentiated neoplasia.
Nonsteroidal anti-inflammatory drugs and breast cancer risk in the National Institutes of Health–AARP Diet and Health Study
Authors: Gierach Gretchen L , Lacey James V , Schatzkin Arthur , Leitzmann Michael F , Richesson Douglas , Hollenbeck Albert R , Brinton Louise A
Published: April 2008

Introduction

By inhibiting cyclooxygenase-2, nonsteroidal anti-inflammatory drugs (NSAIDs) decrease aromatase activity and might reduce breast cancer risk by suppressing estrogen synthesis. Epidemiologic evidence for a protective role of NSAIDs in breast cancer, however, is equivocal.

Methods

We tested NSAID use for its association with breast cancer incidence in the National Institutes of Health–AARP Diet and Health Study, where 127,383 female AARP (formerly known as the American Association of Retired Persons) members with no history of cancer, aged 51 to 72 years, completed a mailed questionnaire (1996 to 1997). We estimated relative risks of breast cancer for NSAID exposures using multivariate Cox proportional hazards regression models. The state cancer registry and mortality index linkage identified 4,501 primary incident breast cancers through 31 December 2003, including 1,439 estrogen receptor (ER)-positive cancers and 280 ER-negative cancers.

Results

Proportional hazards models revealed no statistically significant association between overall NSAIDs and total breast cancer. As cyclooxygenase inhibition by aspirin (but not other NSAIDs) is irreversible, we tested associations by NSAID type. Although we observed no significant differences in risk for daily use (versus nonuse) of aspirin (relative risk = 0.93, 95% confidence interval = 0.85 to 1.01) or nonaspirin NSAIDS (relative risk = 0.96, 95% confidence interval = 0.87 to 1.05), risk of ER-positive breast cancer was significantly reduced with daily aspirin use (relative risk = 0.84, 95% confidence interval = 0.71 to 0.98) – a relationship not observed for nonaspirin NSAIDS. Neither aspirin nor nonaspirin NSAIDs were associated with risk of ER-negative breast cancer.

Conclusion

Breast cancer risk was not significantly associated with NSAID use, but daily aspirin use was associated with a modest reduction in ER-positive breast cancer. Our results provide support for further evaluating relationships by NSAID type and breast cancer subtype.
Improved outcomes from the administration of progesterone for patients with acute severe traumatic brain injury: a randomized controlled trial
Authors: Xiao Guomin , Wei Jing , Yan Weiqi , Wang Weimin , Lu Zhenhui
Published: April 2008

Background

Severe traumatic brain injury (TBI) has been increasing with greater incidence of injuries from traffic or sporting accidents. Although there are a number of animal models of TBI using progesterone for head injury, the effects of progesterone on neurologic outcome of acute TBI patients remain unclear. The aim of the present clinical study was to assess the longer-term efficacy of progesterone on the improvement in neurologic outcome of patients with acute severe TBI.

Methods

A total of 159 patients who arrived within 8 hours of injury with a Glasgow Coma Score ≤ 8 were enrolled in the study. A prospective, randomized, placebo-controlled trial of progesterone was conducted in the Neurotrauma Center of our teaching hospital. The patients were randomized to receive either progesterone or placebo. The primary endpoint was the Glasgow Outcome Scale score 3 months after brain injury. Secondary efficacy endpoints included the modified Functional Independence Measure score and mortality. In a follow-up protocol at 6 months, the Glasgow Outcome Scale and the modified Functional Independence Measure scores were again determined.

Results

Of the 159 patients randomized, 82 received progesterone and 77 received placebo. The demographic characteristics, the mechanism of injury, and the time of treatment were compared for the two groups. After 3 months and 6 months of treatment, the dichotomized Glasgow Outcome Scale score analysis exhibited more favorable outcomes among the patients who were given progesterone compared with the control individuals (P = 0.034 and P = 0.048, respectively). The modified Functional Independence Measure scores in the progesterone group were higher than those in the placebo group at both 3-month and 6-month follow-up (P < 0.05 and P < 0.01). The mortality rate of the progesterone group was significantly lower than that of the placebo group at 6-month follow-up (P < 0.05). The mean intracranial pressure values 72 hours and 7 days after injury were lower in the progesterone group than in the placebo group, but there was no statistical significance between the two groups (P > 0.05). Instances of complications and adverse events associated with the administration of progesterone were not found.

Conclusion

Our data suggest that acute severe TBI patients with administration of progesterone hold improved neurologic outcomes for up to 6 months. These results provide information important for further large and multicenter clinical trials on progesterone as a promising neuroprotective drug.

Trial Registration

ACTRN12607000545460.