Biomedical subjects

M Morgante

Publications and source records attributed to M Morgante.

27 records · Page 2Linked to original sources

PubMed1996

Total and differential cell count by direct microscopic method on ewe milk.

On 700 milk samples from single half udders of Comisana ewes, somatic cell count (SCC) and differential cell count (DCC) were determined, using a Fossomatic 90 cell counter (Foss Electric, Denmark) (SCCF) and milk smears stained with May Grünwald-Giemsa (DCCS). SCC and DCC were also determined with modified KOVAH SYSTEM (Hicor Biomedical Inc. Irvine, CA, USA) (SCCK and DCCK, respectively). Out of 665 milk samples from half udders without clinical signs of mastitis, 640 (Class I) were sterile, while 25 (Class II) were bacteriologically positive. Out of 35 milk samples (Class III) from half udders with clinical signs of mastitis, 25 were bacteriologically positive. Mean results (after logarithmic transformation of cells/ml/10(3)) of SCCF and SCCK for all the 700 milk samples were 1.89 +/- 0.58 and 1.86 +/- 0.60 with linear correlation coefficient (r) of 0.960, while least squares means for Class I, II and III were 1.78, 2.23 and 3.73 respectively and 1.75, 2.19 and 3.74 with r of 0.894, 0.979 and 0.987. Mean results of DCCS and DCCK were 38.1 +/- 23.3, 34.9, 52.1, and 82.2 PMNL% and 41.8 +/- 21.7, 38.6, 60.2, and 87.3 PMNL% with r of 0.855, 0.812, 0.697 and 0.805. The results showed high correlation coefficients and a good reliability between SCCK and SCCF and high correlation coefficients for DCC methods. In conclusion, it could be suggested that the possibility of routine use of the KOVAH SYSTEM method is particularly useful in detecting if an abnormal SCC is due to a polymorphonuclear neutrophil leukocytes increase.

Animals↗

PubMed1995

Hypervariable microsatellites provide a general source of polymorphic DNA markers for the chloroplast genome.

BACKGROUND: The study of plant populations is greatly facilitated by the deployment of chloroplast DNA markers. Asymmetric inheritance, lower effective population sizes and perceived lower mutation rates indicate that the chloroplast genome may have different patterns of genetic diversity compared to nuclear genomes. Convenient assays that would allow intraspecific chloroplast variability to be detected are required. RESULTS: Eukaryote nuclear genomes contain ubiquitous simple sequence repeat (microsatellite) loci that are highly polymorphic in length; these polymorphisms can be rapidly typed by the polymerase chain reaction (PCR). Using primers flanking simple mononucleotide repeat motifs in the chloroplast DNA of annual and perennial soybean species, we demonstrate that microsatellites in the chloroplast genome also exhibit length variation, and that this polymorphism is due to changes in the repeat region. Furthermore, we have observed a nonrandom geographic distribution of variations at these loci, and have examined the number and location of such repeats within the chloroplast genomes of other species. CONCLUSIONS: PCR-based analysis of mononucleotide repeats may be used to detect both intraspecific and interspecific variability in the chloroplast genomes of seed plants. The analysis of polymorphic microsatellites thus provides an important experimental tool to examine a range of issues in plant genetics.

Base Sequence↗

PubMed1995

Polymorphic simple sequence repeat regions in chloroplast genomes: applications to the population genetics of pines.

Simple sequence repeats (SSRs), consisting of tandemly repeated multiple copies of mono-, di-, tri-, or tetranucleotide motifs, are ubiquitous in eukaryotic genomes and are frequently used as genetic markers, taking advantage of their length polymorphism. We have examined the polymorphism of such sequences in the chloroplast genomes of plants, by using a PCR-based assay. GenBank searches identified the presence of several (dA)n.(dT)n mononucleotide stretches in chloroplast genomes. A chloroplast (cp) SSR was identified in three pine species (Pinus contorta, Pinus sylvestris, and Pinus thunbergii) 312 bp upstream of the psbA gene. DNA amplification of this repeated region from 11 pine species identified nine length variants. The polymorphic amplified fragments were isolated and the DNA sequence was determined, confirming that the length polymorphism was caused by variation in the length of the repeated region. In the pines, the chloroplast genome is transmitted through pollen and this PCR assay may be used to monitor gene flow in this genus. Analysis of 305 individuals from seven populations of Pinus leucodermis Ant. revealed the presence of four variants with intrapopulational diversities ranging from 0.000 to 0.629 and an average of 0.320. Restriction fragment length polymorphism analysis of cpDNA on the same populations previously failed to detect any variation. Population subdivision based on cpSSR was higher (Gst = 0.22, where Gst is coefficient of gene differentiation) than that revealed in a previous isozyme study (Gst = 0.05). We anticipate that SSR loci within the chloroplast genome should provide a highly informative assay for the analysis of the genetic structure of plant populations.

Base Sequence↗

PubMed1995

Paternal inheritance of plastids in interspecific hybrids of the genus Actinidia revealed by PCR-amplification of chloroplast DNA fragments.

RFLPs (restriction fragment length polymorphisms) of PCR (polymerase chain reaction) -amplified fragments were used to trace the pattern of plastid DNA inheritance in the genus Actinidia. A total of 51 progeny originating from interspecific crosses between three A. arguta cultivars and A. deliciosa, the kiwifruit, and 12 progeny originating from the cross between A. kolomikta and A. chinensis were analysed together with their parents. No reciprocal crosses could be tested since they all failed to set viable seeds. Attempts to rescue immature embryos failed in all cases as well. The A. argutaXA. deliciosa crosses were checked for the RFLP patterns of a sequence encoding part of the Rubisco large subunit (rbcL), using either AluI or MseI, and for a sequence encoding part of the photosystem II D1 protein (psbA), using HinfI. The A. kolomiktaXA. chinensis cross was checked for the RFLP patterns of sequences encoding the spacers between trnT and the 5'-trnL exon (a-b spacer DNA) and the trnL 3' exon and trnF (e-f spacer DNA), respectively. The first spacer revealed a natural polymorphism between the two parent species due to a large deletion occurring in A. kolomikta detectable without further restriction enzyme treatment. The e-f spacer DNA was digested with HinfI. The comparison of the RFLP patterns in the parents and their progeny showed a strictly paternal inheritance of chloroplast DNA in Actinidia, with no exception found in any of the crosses examined. As the reciprocal crosses were not available, we do not know whether paternal inheritance of plastids is restricted to the crosses we analysed or if this is the general rule for plastid inheritance in the genus Actinidia. Actinidia is dioecious and is the first purely outbreeding species for which a paternal plastid inheritance has so far been documented.

Base Sequence↗

PubMed1994

Allozyme and chloroplast DNA variation in Italian and Greek populations of Pinus leucodermis.

Allozyme and chloroplast (cpDNA) variation was examined in five Italian and two Greek populations of Pinus leucodermis Ant. to estimate levels of genetic variability within and among populations and to establish the usefulness of allozyme and cpDNA markers in the taxonomic classification of these populations. Twenty-three isozyme gene loci were analysed, as well as restriction fragment length polymorphisms at two cpDNA spacer regions between tRNA genes. The level of genetic variability tended to be lower in the Greek populations but overall the observed levels of allozyme variation within and among populations were similar to those of other conifers. Identical cpDNA amplification and restriction patterns were observed among all individuals sampled from the seven populations. Taken together, the results of the allozyme and cpDNA analyses indicate that all seven populations belong to the same biological species.

Alleles↗

PubMed1994

Genetic mapping and variability of seven soybean simple sequence repeat loci.

Microsatellites or simple sequence repeats are stretches of short tandemly repeated DNA sequence motifs, dispersed throughout the genomes of most eukaryotes. Simple sequence repeat polymorphisms (SSRPs) have recently been reported in plants. Here we present the genetic map position of seven different soybean (Glycine max (L.) Merr. and Glycine soja Sieb. and Zucc.) SSRPs contained in sequenced genes, four of which represent newly mapped positions for these genes. The other three SSRPs coincided with independently established RFLP map positions for the corresponding genes. When a set of 61 soybean accessions was screened at four of these loci by using agarose gels, the average number of alleles per locus was 7.75, the effective number of alleles (ne) was 2.57, and the level of allele differentiation (delta(t)) was 0.62. Allelic variation decreased sharply with increasing levels of domestication, with the level of differentiation going from 84% in the wild soybean to 43% in the elite germplasm. Variation levels observed on a subset made of 19 of the 61 lines were always higher for SSRPs than for RFLP markers, with the average number of alleles per locus going from 4.25 to 2.15. In comparison with RFLP markers, SSRPs are more informative and are easier to analyse but require more effort to develop.

Base Sequence↗

PubMed1993

PCR-amplified microsatellites as markers in plant genetics.

In order to assess the feasibility of using microsatellites as markers in plant genetics, a survey of published DNA sequence data for presence, abundance and ubiquity in higher plants of all types of dinucleotide and trinucleotide repeats with a minimum number of 10 and 7 units, respectively, was conducted. This search revealed that such microsatellites are frequent and widely distributed; they were uncovered in 34 species, with a frequency of one every 50 kb. AT repeats were by far the most frequently observed class of dinucleotide microsatellites, whereas AC/TG repeats, which are common in animals, were observed only once. TAT repeats prevailed among trinucleotides. Polymerase chain reaction amplification of (AT)n and (TAT)n microsatellites in soybean (Glycine max (L.) Merr.) revealed that they are highly polymorphic, as a consequence of length variation, somatically stable and inherited in a co-dominant Mendelian manner. The abundance and amount of information derived from such markers, together with the ease by which they can be identified, make them ideal markers for plant genetic linkage and physical mapping, population studies and varietal identification.

Base Sequence↗

PubMedDate not supplied

Soybean chromosome painting: a strategy for somatic cytogenetics.

Cytological identification of soybean mitotic metaphase chromosomes (2n = 40) has been severely limited by their small size and uniform karyomorphology. We have developed fluorescent in situ hybridization (FISH), PCR-primed in situ labelling (PCR-PRINS) procedures, and molecular probes for routine cytological identification and for the physical mapping of soybean somatic chromosomes. Chromosome preparation has been achieved by modifications of previous protocols and through the preparation of root-tip protoplasts prior to chromosome spreading. Initially our probe selection focused on highly repeated DNAs that provide very intense localized hybridization signals. Repetitive gene probes that have proven valuable include the rDNA loci (5S and 45S) which are chromosome specific. We have also developed satellite DNA probes for two different sequence families: the SB92 and the STR120 satellites. Both of these are tandemly arranged at multiple chromosomal loci. By using different cloned examples of each family, we have been able to selectively label unique subsets of soybean chromosomes. Double hybridization with biotin and digoxigenin labeled probes has allowed us to determine the chromosomal overlap between different probes. In addition, we have joined portions of the metaphase chromosome painting patterns with the genetic map by single-copy FISH and PCR-PRINS detection of the RFLP loci G8.15, G17.3, and A199a and A199b. Total genomic DNA in situ hybridization (GISH) patterns were also used to characterize the soybean chromosomes.

Chromosome Mapping↗

PubMedDate not supplied

[YKL 40: marker of disease activity in rheumatoid arthritis?].

In patients with rheumatoid arthritis the serum and synovial fluid levels of a glycoprotein called YKL 40 are correlated to the severity of disease. YKL 40 may be related to disease activity in RA; it is induced by vitamine D and is reduced by TGF b. YKL 40 has been isolated from bovine mammary secretions during non lactating period and from Human osteosarcoma cell line MG-63. Some observations on experimental and clinical studies are presented.

Adipokines↗