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PubMed1995

Heritability and selection on body size in a natural population of Drosophila buzzatii.

An attempt was made to assess whether the phenotypic differences in body size (as measured by wing length) between wild-caught mating and single Drosophila buzzatii males could be attributed to genetic differences between the samples. Mating males were found to be larger and less variable than a random sample of the population. The progeny of the mating males (produced by crossing to a random female from a stock derived from the same population) were on average larger than those of the single males, but not significantly so (P = 0.063), and less phenotypically variable. This difference in variance between the samples suggests that there are indeed genetic differences between the paternal samples but tests for significant differences in the additive genetic component of variance proved inconclusive. For both samples it was found that while the ratio of additive genetic variation in the laboratory to phenotypic variation in the field yielded estimates of ĥs2(N) congruent to 10% the regression of offspring reared in the laboratory on parents from the wild was not significantly different from zero. In addition, it was found that the average development time of the progeny of the mating males is shorter than that of the random sample.

Analysis of Variance↗

PubMed1996

Sarcomatoid carcinomas: pathological and histopathogenetic considerations.

Malignant tumors with a mixed phenotype are a controversial field of pathology. In this article the morphological aspects and the immunohistological characterization of sarcomatoid carcinomas are presented. These uncommon neoplasms show both carcinomatous and sarcomatous features, and have been described in the past under a variety of different names causing great uncertainty about their classification and histogenesis. They can occur in various anatomical sites and exhibit a wide range of microscopic appearances, but some features are quite characteristic and are found in many cases. Morphological "transition" between carcinomatous and sarcomatous tissue, and detection of epithelial characteristics by electron microscopy or immunohistochemistry in the sarcomatous component, are very peculiar features of these neoplasms, providing both helpful clues for pathological diagnosis and important insights into histogenesis. Here a unifying histopathogenetic mechanism based on the phenotypic conversion of carcinoma into sarcomatoid tissue is proposed and supporting literature data from both experimental systems and clinicopathological observations are reviewed and discussed.

Carcinoembryonic Antigen↗

PubMed1992

Phenotypes of conjunctival inflammatory cells in sarcoidosis.

Phenotypes of the infiltrating mononuclear cells of the lower fornix conjunctiva of nine patients with sarcoidosis and six controls were studied using monoclonal antibodies and a modified immunoperoxidase method. Four patients had sarcoidosis of recent onset (duration of 2 years or less) and five patients had a chronic disease (duration of 3 or more years). The inflammatory cells in the sarcoid conjunctival specimens were predominantly T lymphocytes, the vast majority of which were of T helper/inducer subtype expressing Leu-3a + 3b positivity. The ratio of T helper/inducer cells to T suppressor/cytotoxic cells was 3.9 on average but only 0.9 in controls. Epithelioid cell granulomas were seen in three specimens in one case of recent onset and in two chronic cases comprising a marked amount (more than 15 cells/visual field) of cells bearing phenotypes of macrophages, T cells, T helper/inducer cells and HLA-DR antigen, and in smaller quantities of T suppressor/cytotoxic cells. The mean number of all immunocompetent cell subtypes of specimens from newly diagnosed patients exceeded that of specimens from chronic patients. We believe that the sarcoid immune reaction in the conjunctiva is a dynamic process in which proliferation of immunocompetent mononuclear cells precedes the stage of granuloma formation.

Adult↗

PubMed2002

Molecular characterization of cytoplasmic and nuclear genomes in phenotypically abnormal Valencia orange (Citrus sinensis) + Meiwa kumquat (Fortunella crassifolia) intergeneric somatic hybrids.

Organelle DNA inheritance of four 10-year-old somatic hybrid trees between Valencia orange [Citrus sinensis (L.) Osbeck] and Meiwa kumquat (Fortunella crassifolia Swingle) was analyzed by cleaved amplified polymorphic sequence (CAPS) and restriction fragment length polymorphisms (RFLPs). Five chloroplast (cp) and three mitochondrial (mt) universal primer pairs were amplified, but no polymorphisms were detected. When the polymerase chain reaction products were digested by 15 restriction enzymes, four polymorphic cpDNA-CAPS and two mtDNA-CAPS markers were found. Both the cpDNA and mtDNA in the somatic hybrids were derived from Valencia orange (the embryogenic suspension parent). Genomic DNA of the somatic hybrids and corresponding parents was digested by five restriction endonucleases and hybridized with one chloroplast probe (RbcL- RbcL) and nine mitochondrial probes (coxI, coxII, c oxIII, c ob, atpA, tyr, proI, atp6 and atp9). The results indicated that three hybrid plants shared one strong cpDNA band with both parents and that the remaining one plant had two additional novel bands besides the shared band, while their mtDNA was identical to that of Valencia orange plus non-parental bands. When data on the mtDNA banding patterns were combined with observations on phenotypic performance in the field, it was found that the more complex mtDNA banding pattern coincided with increased vigor of the plant. The stability of the organelle genomes was studied by extracting the genomic DNA of one hybrid plant at monthly intervals for 1 year and then analyzing it using RFLPs. Before the dieback of the shoots, two fragments of the mtDNA were lost while the cpDNAs remained stable. Ploidy analysis by flow cytometry showed that all of the hybrids were stable tetraploids. Four simple sequence repeat primer pairs were applied to detect microsatellite alleles of the four hybrid plants, both parents and the 12 DNA samples from one plant. The results showed that all hybrids had biparental bands uniformly, which indicated that they had the same nuclear background. These results suggest that the mtDNA pattern is correlated with the phenotypic abnormality of Valencia and kumquat somatic hybrid plants and that nuclear-cytoplasm incompatibility may be the cause of dieback.

Cell Nucleus↗

PubMed2006

Exaggerated behavioral phenotypes in Fmr1/Fxr2 double knockout mice reveal a functional genetic interaction between Fragile X-related proteins.

Individuals affected by Fragile X syndrome (FXS) experience cognitive impairment, hyperactivity, attention deficits, social anxiety and autistic-like behaviors. FXS results from the loss of expression of the Fragile X mental retardation (FMR1) gene, whose protein product FMRP is thought to play an important role in neuronal function and synaptic plasticity. Two paralogs of FMRP, FXR1P and FXR2P, have been identified, forming the Fragile X-related (FXR) family of proteins. Although the functions of FXR1P and FXR2P are not well understood, there are similarities among all three FXR proteins in gene structure, amino acid sequence, expression pattern and cellular functions. Mouse models have been described for loss of Fmrp, Fxr1p and Fxr2p, the mouse homologs of FMRP, FXR1P and FXR2P. In earlier studies, we found that Fmr1 knockout (KO) mice, which do not express Fmrp, and Fxr2 KO mice, which do not express Fxr2p, show similarities in some behavioral responses such as hyperactivity. To better understand the functional relationship between FMRP and FXR2P, we generated Fmr1 KO, Fxr2 KO, Fmr1/Fxr2 double KO and wild-type control mice as littermates on the same genetic background and examined them in several behavioral assays. Results show that Fmr1/Fxr2 double KO mice have exaggerated behavioral phenotypes in open-field activity, prepulse inhibition of acoustic startle response and contextual fear conditioning when compared with Fmr1 KO mice, Fxr2 KO mice or wild-type littermates. Our findings suggest that Fmr1 and Fxr2 genes contribute in a cooperative manner to pathways controlling locomotor activity, sensorimotor gating and cognitive processes.

Acoustic Stimulation↗

PubMed2000

Characterization of epidemic and nonepidemic methicillin-resistant Staphylococcus aureus strains in a university hospital in northeast Germany.

The aim of this study was to analyze methicillin-resistant Staphylococcus aureus strains isolated during a 1-year period by means of pulsed-field gel electrophoresis, resistance phenotyping and determination of biochemical features. Eight different resistance phenotypes with the predominant resistance type Pen Oxa Cip (penicillin, oxacillin, ciprofloxacin) were observed. None of the strains tested exhibited decreased susceptibility to vancomycin, but two strains were resistant to mupirocin. Genetic relatedness of methicillin-resistant Staphylococcus aureus isolates could be shown for two outbreaks, one of which was caused by a clone with an epidemic potential concerning duration of colonization/infection of patients and dissemination of the strains in the hospital.

Anti-Bacterial Agents↗

PubMed1997

Genomic homogeneity of the AHU/IA-1,2 phenotype of Neisseria gonorrhoeae during its disappearance from an urban population.

BACKGROUND: Isolates of Neisseria gonorrhoeae requiring arginine, hypoxanthine, and uracil (AHU) appeared in Denmark in 1946, were preponderant in Seattle during the 1970s, were associated with disseminated gonococcal infections (DGI), and were primarily of the IA-1,2 serovar. GOALS: To investigate the disappearance of the AHU/IA-1,2 phenotype and to examine by pulsed-field gel electrophoresis (PFGE) the genomic homogeneity of this unique phenotype isolated from Seattle-King County during the past decade. STUDY DESIGN: This retrospective study used data extracted from previous publications for the period 1971 through 1984, and from existing records at the Neisseria Reference Laboratory, University of Washington for the period 1985 through 1996. Samples (n = 68) of AHU/IA-1,2 isolates from 1984 to 1985 and 1988 to 1993 were analyzed using endonucleases NheI and XbaI. For comparison, 10 AHU/IB isolates were included in the study. RESULTS: AHU isolates, predominantly IA-1,2 strains accounted for 52% of the gonococcal isolates for the period 1971 through 1974, 40% for 1974 through 1976, 16% for 1984, and then declined from 7% in 1986 to 0% for each of the last 3 years, 1994 through 1996. Isolates with < or = 1 band difference after digestion with either NheI or XbaI were considered to belong to a single closely related pattern. Pulsed-field gel electrophoresis (PFGE) type designation was made from the combination of NheI and XbaI patterns. These criteria yielded 5 NheI and 8 XbaI PFGE patterns, and 11 PFGE types based on the combination of NheI and XbaI pattern. The most frequently occurring NheI and XbaI patterns accounted for 74% and 57% of isolates, respectively. One type persisted throughout the decade and accounted for 54% of the 68 isolates. Analysis of the 10 AHU/IB isolates yielded 7 NheI and 8 XbaI patterns that gave 9 types that were distinct from the types found in the AHU/IA-1,2 strains. CONCLUSION: The AHU/IA-1,2 phenotype first documented 50 years ago in Denmark still shows a high degree of genomic homogeneity during the past decade in Seattle. The implementation of screening, decreased rates of sexual exposure in the acquired immune deficiency syndrome era, or other factors may explain its apparent elimination in Seattle-King County.

Electrophoresis, Gel, Pulsed-Field↗

PubMed1999

Plasmodium falciparum field isolates commonly use erythrocyte invasion pathways that are independent of sialic acid residues of glycophorin A.

Erythrocyte invasion by malaria parasites is mediated by specific molecular interactions. Sialic acid residues of glycophorin A are used as invasion receptors by Plasmodium falciparum. In vitro invasion studies have demonstrated that some cloned P. falciparum lines can use alternate receptors independent of sialic acid residues of glycophorin A. It is not known if invasion by alternate pathways occurs commonly in the field. In this study, we used in vitro growth assays and erythrocyte invasion assays to determine the invasion phenotypes of 15 P. falciparum field isolates. Of the 15 field isolates tested, 5 multiply in both neuraminidase and trypsin-treated erythrocytes, 3 multiply in neuraminidase-treated but not trypsin-treated erythrocytes, and 4 multiply in trypsin-treated but not neuraminidase-treated erythrocytes; 12 of the 15 field isolates tested use alternate invasion pathways that are not dependent on sialic acid residues of glycophorin A. Alternate invasion pathways are thus commonly used by P. falciparum field isolates. Typing based on two polymorphic markers, MSP-1 and MSP-2, and two microsatellite markers suggests that only 1 of the 15 field isolates tested contains multiple parasite genotypes. Individual P. falciparum lines can thus use multiple invasion pathways in the field. These observations have important implications for malaria vaccine development efforts based on EBA-175, the P. falciparum protein that binds sialic acid residues of glycophorin A during invasion. It may be necessary to target parasite ligands responsible for the alternate invasion pathways in addition to EBA-175 to effectively block erythrocyte invasion by P. falciparum.

Animals↗

PubMed2005

Vaccines for enterotoxigenic Escherichia coli: current status.

PURPOSE OF REVIEW: Enterotoxigenic Escherichia coli (ETEC) remain major causes of infantile diarrhea in the developing world and of travelers' diarrhea in visitors to these areas from industrialized countries, although the relative importance of these pathogens in these settings may be decreasing. The pathogenesis of ETEC infections has been well understood for almost two decades, and there is good evidence for acquired immunity after infection. Nevertheless, currently there is no effective ETEC vaccine. This review evaluates the current status of ETEC vaccine development. RECENT FINDINGS: ETEC organisms express fimbrial (or fibrillar) colonization factor antigens that function as adhesins to promote their attachment to the small intestinal epithelium. They secrete either (or both) of two major protein enterotoxins that induce fluid and electrolyte secretion. Vaccine development during the last decade has targeted the components of the three major colonization factor antigens as well as the immunogenic heat-labile enterotoxin. This strategy was expected to cover 90% of infecting strains, leaving unprotected only those strains without major colonization factor antigens and those that express only the poorly immunogenic heat-stable enterotoxin. Recent experiences have questioned the validity of the current vaccine strategy since new reports indicate that (1) the number of recognized colonization factor antigens of ETEC has increased to more than 21, (2) epidemiologic field studies of children in endemic areas suggest that infection with ETEC of a given colonization factor antigen/toxin phenotype may not confer protection on reinfection with other strains of the same colonization factor antigen/toxin phenotype, and (3) a major field trial of a heat-killed ETEC vaccine expressing colonization factor antigens and containing the B subunit of cholera toxin as a surrogate for E. coli heat-labile enterotoxin was ineffective against ETEC infections that should have been "vaccine preventable." New vaccine strategies have been developed to deliver ETEC antigens to the mucosal immune system. SUMMARY: Although new vaccines are being developed to improve immunogenicity over that of the heat-killed vaccine, the current strategy for antigen inclusion has been challenged and new, common antigens may have to be defined to achieve the goal of an effective vaccine against ETEC.

Bacterial Toxins↗

PubMed1987

"Unclassifiable" weak A blood group and deficient H phenotype (Hm) in one pedigree. Variants of A and H types in a family.

A new variant of blood group A [A(WAS)] was expressed in three generations of a Caucasian family: Phenotype included weak mixed field hemagglutination by anti-A reagents, secretion of H substance, and presence of anti-A1 in serum. The A(WAS) variant was inherited in a Mendelian fashion, dominant to O. A-transferase activity was absent from cells and saliva but was 0.2% of normal A1 transferase activity in serum, with a pH optimum of 6.0. Family members expressing A(WAS) also demonstrated partly deficient H type on cells (Hm). H-transferase activity in serum was normal for a weak A subgroup and showed typical Km and acceptor specificities. Linkage of H-modifier and ABO loci cannot be excluded.

ABO Blood-Group System↗

PubMed1996

Molecular polymorphism and phenotypic variation in Aspergillus carbonarius.

Thirteen collection strains and field isolates of Aspergillus carbonarius were examined by using various genotypic and phenotypic approaches. Restriction fragment length polymorphism analysis of the ribosomal RNA gene cluster and the mitochondrial DNA of the strains revealed only slight variations, except for one field isolate (IN7), which exhibited completely different ribosomal RNA gene cluster and mitochondrial DNA patterns. The mitochondrial DNAs of these strains were found to be much larger (45 to 57 kb) than those found earlier in the A. niger aggregate. Strain-specific characters could be detected by the random amplified polymorphic DNA technique. Isoenzyme analysis and examination of carbon source utilisation patterns of the strains also revealed some intraspecific variability, though much smaller than that observed by using DNA-based techniques. The dendrograms constructed based on genotypic and phenotypic data suggest that strain IN7 might represent a new subspecies of A. carbonarius.

Acid Phosphatase↗

PubMed1994

Quantitative phenotyping of N'Dama cattle for aspects of trypanotolerance under field tsetse challenge.

Matching animal health and performance data were recorded over the 2 year period from weaning at 10 months of age on 255 N'Dama cattle in a high natural tsetse challenge situation in Zaire. Four parameters that are regarded as possible indicators of trypanotolerance, species of trypanosomes detected, length of time parasitaemic, intensity of parasitaemia (parasitaemia score), and anaemic condition as estimated by packed cell volume (PCV) values, were measured and the relative effects of changes in these parameters on trypanocidal drug requirements and growth were assessed. The effects of species of trypanosome on drug requirements and growth were directly measurable. In the case of the other three indicators, the effects on drug requirements and growth that would be brought about by a change of one standard deviation in each were calculated. This allowed comparison of similar sized changes in these three indicators that are of necessity recorded in dissimilar units. Trypanosoma vivax and Trypanosoma congolense infections had equal effects on the number of trypanocidal drug treatments required, an average of 0.61 treatments being administered to each infected post-weaner. A reduction of one standard deviation (SD) in length of time infected reduced the number of treatments required by 0.23 or 36% and an increase of 1 SD in PCV reduced the number required by 0.27 or 43%. Changes in parasitaemia score were not important. In the case of growth, a T. congolense infection reduced growth by 12.4 g day-1 or 8% more than a T. vivax infection. A reduction of 1 SD in length of time infected increased growth by 9.8 g day-1 or 6.5%, a reduction of 1 SD in parasitaemia score increased growth by 9.0 g day-1 or 6.0%, and an increase of 1 SD in average PCV increased growth by 8.4 g day-1 or 5.6%. The necessity to simultaneously measure the four criteria is clearly indicated by their approximately equal effects on the final performance trait of daily liveweight gain. Thus, absence of information on any of these criteria would significantly affect the accuracy of the estimate of an animal's overall trypanotolerance phenotype in this central African situation and reduce the progress possible in production projects involving N'Dama cattle.

Anemia↗

PubMedDate not supplied

Correlation among phenotypical and molecular techniques in comparing ascomycetous yeast type strains.

Different phenotypical or molecular techniques can be used to describe and classify microorganisms for taxonomic, phylogenetic or genetic purposes. In yeast taxonomy the official hierarchic classification, based on morphological and physiological characters, is used together with more convenient molecular techniques such as the DNA sequencing. The question on whether these procedures produce coherent classifications is critical both to interpret taxonomic data consistently and to outline species correctly. In this paper, a set of type strains from the major genera of the budding hemiascomycetes yeast is examined with a series of physiological and molecular techniques, widely employed in taxonomy, in order to compare the among-strains correlations obtained with different methods. Results showed that the level of correlation among different techniques is relatively low, showing that different classifications and species organization could be obtained with diverse approaches. This is particularly interesting, considering that the official description of the yeast species is based on characters different from those becoming increasingly popular in the routine identification.

Animals↗

PubMed1999

Invasive and noninvasive group A streptococcal isolates with different speA alleles in The Netherlands: genetic relatedness and production of pyrogenic exotoxins A and B.

Streptococcal pyrogenic exotoxin A (SPE-A) and SPE-B have been implicated in the pathogenesis of severe group A streptococcal (GAS) disease. We studied 31 invasive GAS strains including 18 isolates from patients with toxic shock syndrome and 22 noninvasive strains isolated in The Netherlands between 1994 and 1998. These strains were associated with the different allelic variants of the gene encoding SPE-A. We selected endemic strains with speA-positive M and T serotypes: speA2-associated M1T1 and M22-60T12 strains, speA3-associated M3T3 strains, and speA4-associated M6T6 strains. Since speA1-positive isolates were not frequently encountered, we included speA1 strains of different serotypes. The GAS strains were compared genotypically by pulsed-field gel electrophoresis and phenotypically by the in vitro production of SPE-A and SPE-B. All strains within one M and T type appeared to be of clonal origin. Most strains produced SPE-A and SPE-B, but only a minority of the speA4-positive isolates did so. Among our isolates, speA1- and speA3-positive strains produced significantly more SPE-A than speA2- and speA4-carrying strains, while SPE-B production was most pronounced among speA1- and speA2-containing strains. There was a marked degree of variability in the amounts of exotoxins produced in vitro by strains that shared the same genetic profile. We conclude that the differences in the in vitro production of SPE-A and SPE-B between our selected strains with identical M and T types were not related to either genetic heterogeneity or the clinical course of GAS disease in the patient from whom they were isolated.

Alleles↗

PubMed2005

Molecular and phenotypical characterization of Clostridium perfringens isolates from poultry flocks with different disease status.

Due to the diminished use of growth-promoting antibiotics in the European Union, Clostridium perfringens induced necrotic enteritis and subclinical disease have become important threats to poultry health. A study was set up to genotypically and phenotypically characterise C. perfringens isolates from poultry flocks with different health status. Animals from healthy flocks were sampled by cloacal swabs, while intestinal and liver samples of animals suffering from necrotic enteritis were analysed. A total of 27 isolates was obtained from 23 broiler flocks without clinical problems and 36 isolates were obtained from 8 flocks with clinical problems. Using PFGE typing, high genetic diversity was detected between isolates from different flocks. Isolates derived from flocks where disease outbreaks occurred were clonal within each flock, but each flock harboured a different clone. All isolates were of toxin type A. Isolates from 5 out of 35 PFGE types carried the cpb2 gene, encoding the beta2 toxin, and isolates from 2 out of 35 PFGE types harboured the cpe gene, encoding the enterotoxin. In vitro alpha toxin production for all isolates was quantified by enzyme-linked immunosorbent assay. It was shown that in vitro alpha toxin production of C. perfringens isolates from diseased flocks was not higher than in vitro alpha toxin production from isolates derived from healthy flocks.

Animals↗

PubMed2000

Gliding mutants of Mycoplasma mobile: relationships between motility and cell morphology, cell adhesion and microcolony formation.

The present study characterizes gliding motility mutants of Mycoplasma mobile which were obtained by UV irradiation. They were identified by their abnormal colony shapes in 0.1% agar medium, showing a reduced number of satellite colonies compared to the wild-type. A total of ten mutants were isolated based on their colony phenotype. Using dark-field and electron microscopy, two classes of mutants, group I and group II, were defined. Cells of group I mutants had irregular, flexible and sometimes elongated head-like structures and showed a tendency to aggregate. Neither binding to glass nor gliding motility was observed in these mutants. Cells of group II mutants were rather spherical in shape, with the long axis reduced to 80% and the short axis enlarged to 120% of that of wild-type cells, respectively. Their gliding speed was 20% faster than that of wild-type cells. Three of the ten mutants remained unclassified. Mutant m6 had a reduced binding activity to glass and a reduced gliding motility with 50% of the speed of the wild-type strain. The ability of wild-type and mutant colonies to adsorb erythrocytes was found to correlate with the binding activity required for gliding, indicating that mycoplasma gliding depends on cytadherence-associated components. Finally, the ability to form microcolonies on surfaces was shown to correlate with the gliding activity, suggesting a certain role of gliding motility in the parasitic life-cycle of mycoplasmas.

Animals↗

PubMed1995

Variation in tolerance to Bacillus thuringiensis among and within populations of the spruce budworm (Lepidoptera: Tortricidae) in Ontario.

Variation in tolerance to Bacillus thuringiensis Berliner subsp. kurstaki (strain HD-1-S-1980) among and within populations of the spruce budworm, Choristoneura fumiferana (Clemens), was assessed in the laboratory. Force-feeding assays using offspring of females collected as pupae from nine locations throughout Ontario and from a laboratory colony (DCF) demonstrated limited variation in tolerance among populations. Variation among populations was comparable with the variation observed among repeated assays with different batches of larvae from the DCF colony. Population LC50s were not significantly associated with age of the outbreak, host-plant species, incidence of the microsporidian Nosema fumiferanae (Thomson), or size of the female parent. Upper limits for genetic variation in tolerance were estimated by examining variation among full-sibling families within same populations. Mortality of individual families ranged from 6.5 to 70.9% within five field populations and from 2.7 to 93.3% within two laboratory colonies in response to a dose that caused a mean mortality of 40%. Familial factors accounted for 32.8% of the phenotypic variation in response across field populations, as compared with 3% for population factors. These data suggest that the phenotypic variation in tolerance to B. thuringiensis has a substantial genetic component and may provide a basis for evolution of resistance given sufficient selection pressure.

Animals↗

PubMedDate not supplied

From phenotype to genotype.

Advances in the field of genomics have made it possible to decipher the genetic and transcriptional changes that underlie differences among organisms. Here we discuss the merits and drawbacks of these strategies as they relate to evolutionary biology. We suggest that the molecular basis of natural variation can best be interpreted via the intersection of genomic and transcriptome approaches. We outline how this might be accomplished in practice and assemble the components of a method to comprehend the evolutionary interplay between genotype and phenotype.

Biological Evolution↗