Interaction between three types of variation and fluctuating asymmetry in the population of malaria mosquito Anopheles messeae Fall

In the population of the malaria mosquito Anopheles messeae, interrelations of three types of variation (polymorphism for inversions, dorsal color of abdomen and thorax, and number of dents in the crests of the stigmal plate) and fluctuating asymmetry were studied. The presence of white pigmentation on the larval dorsum did not correlate with inversion polymorphism and fluctuating asymmetry, but correlated with the number of small and large dents in the crests of stigmal plate. Asymmetry was shown to have a weak positive correlation with the number of dents. In larvae having a small number of dents, a decrease in the frequency of autosomal homozygotes 3R00 and an increase in the frequency of autosomal heterozygotes 3R01 were found. The dents in the crests of the stigmal plate had two morphological forms: a large one, which is associated with arms 2R and XL, and a small one, which is associated with arms 3R, XL, and 3L. Correlation between inversion polymorphism and bilateral (fluctuating) asymmetry was more complex. The group of individuals with a difference by one dense differed from that with a difference by three or more dents in the frequency of chromosome sequences XL0, XL1, and 2R0. Chromosome sequences XL1, 2R0, 3R0, and 3L1 were classified as those increasing the developmental stability of individuals; XL2, XL0, 2R1, 3R1, and 3L0, as those decreasing it.     

Dialogue medicine: a health-liberating attitude in general practice

The threespine stickleback Gasterosteus aculeatus is an important model in evolutionary and ethologic studies. Its utility would greatly be increased by the availability of molecular markers distinguishing individuals and populations. Such markers can be provided by the major histocompatibility complex (Mhc) genes, which are well known for their extensive polymorphism. In the present study, both class I and class II B Mhc genes have been identified and sequenced. Fifteen distinct class I exon 2 and exon 3 sequences were obtained and assigned to 12 loci on the basis of intron 2 length differences. Some of the loci appear to be related to class I loci identified previously in cichlid fish. The intron 2 sequences and insertions/deletions in exon 2 group the loci into three families (with one family divided further into two subfamilies) derived from different ancestral genes. The ancestors presumably diverged from one another before the divergence of Gasterosteiformes from Perciformes. The 12 distinct class II B sequences may be derived from six loci, which are, however, closely related to one another in both exonic and intronic parts and may have diverged from a single common ancestor after the divergence of Gasterosteiformes from Perciformes. The intron 2 of some of the class I genes contains two microsatellites that can be used as markers, in addition to the polymorphism of the Mhc genes in their exonic regions.     

Comparison of the bactericidal action of amikacin, netilmicin and tobramycin in free and liposomal formulation against Pseudomonas aeruginosa

Glycoprotein Ibalpha (GP Ibalpha; CD 42b; hereafter GPIBA) is a component of the cell surface receptor for the von Willebrand factor (vWf) on platelets. Immunizations against various platelet surface antigens play a major role in neonatal alloimmune thrombocytopenia and in post-transfusion purpura. Only one antigenic polymorphism in GPIBA has thus far been established: the HPA-2 (Ko) alloantigen system. To screen other polymorphisms in GPIBA systematically, we analyzed the whole coding sequence of the GPIBA gene in 50 Finnish blood donors using the single-strand conformation polymorphism method. In addition to the known polymorphisms, we detected three others. Sequencing of the gene segments carrying the new polymorphisms revealed that none of them changed the predicted amino acid sequence. Polymorphism designated RS was located five base pairs upstream from the initiation codon at position 3064 and had the gene frequency of 16% for R and 84% for S, respectively, in the Finnish population, and it was detectable by the restriction enzyme Hae III. The EF polymorphism was at position 3842 (Asn242) and the gene frequencies were 97% for E and 3% for F. The KL polymorphism was at position 4142 (Arg342) and the gene frequencies were 98% for K and 2% for L. The five polymorphic positions in GPIBA formed altogether six different alleles of the gene. The data suggest that there are only a few variable amino acids in GPIBA.     

Human SP-A locus: allele frequencies and linkage disequilibrium between the two surfactant protein A genes

Two surfactant protein A (SP-A) genes and several alleles for each SP-A locus have been previously described. In this report we investigate the potential usefulness of the SP-A loci as markers for genetic studies. We establish conditions that allow the identification of alleles with very similar sequences; We also determine the degree of polymorphism for each SP-A locus: The heterozygosity and polymorphism information content (PIC) values for the SP-A1 locus are 0.63 and 0.55, respectively, and for the SP-A2 locus are 0.50 and 0.56. In the course of these studies, we identify one new SP-A2 allele and show that the SP-A1 and SP-A2 loci are in linkage disequilibrium (P < 0.000001). We also identify 19 of the 20 possible haplotypes in a population of n = 239. Nine of the observed haplotypes reach statistical significance (P < 0.01) in this population, and the segregation of two haplotypes (6A2/1A0 and 6A4/1A) without recombination is verified in a family pedigree. These data together indicate that both SP-A loci are sufficiently polymorphic to be good markers for use in genetic studies. Furthermore, the finding of one novel allele suggests that additional unknown SP-A alleles are yet to be found.     

Restriction fragment length polymorphism of genes of the alpha 2(XI) collagen, bone morphogenetic protein-2, alkaline phosphatase, and tumor necrosis factor-alpha among patients with ossification of posterior longitudinal ligament and controls from the Japanese population

STUDY DESIGN: The present study analyzed the restriction fragment length polymorphism patterns of alpha 2(XI) collagen, bone morphogenetic protein-2, alkaline phosphatase, and tumor necrosis factor-alpha genes in patients with ossification of the posterior longitudinal ligament. This study investigates the genetic polymorphism of bone-induced factors in patients with ossification of the posterior longitudinal ligament and compares it with healthy control subjects. OBJECTIVES: To clarify the genetic markers linked to ossification of the posterior longitudinal ligament. SUMMARY OF BACKGROUND DATA: Ossification of the posterior longitudinal ligament is a genetic disease associated with abnormal calcium metabolism involving the posterior longitudinal ligament. Previous genetic studies have not identified the pathologic mechanism of ossification of the posterior longitudinal ligament. Histopathologic studies of ossification of the posterior longitudinal ligament and the animal model, the spinal hyperostotic mouse, have revealed an increase in Type XI collagen and bone morphogenetic protein-2 expression. METHODS: Eighteen Japanese patients with ossification of the posterior longitudinal ligament and 51 healthy, unrelated control subjects were investigated for the restriction fragment length polymorphism patterns of COL11A2, bone morphogenetic protein-2, alkaline phosphatase, and tumor necrosis factor-alpha, genes with various restriction endonucleases. RESULTS: The gene frequencies of COL11A2 obtained with BamHl (10.0 kb fragment) and HindIII (19.0 kb fragment) observed in patients with ossification of the posterior longitudinal ligament were higher compared with control subjects (0.43 and 0.14, respectively). These differences were statistically significant (BamHl P = 0.018; Hindlll P = 0.046). Two new restriction fragment length polymorphism patterns were detected of the bone morphogenetic protein-2 gene with Mspl and Taql and one already known restriction fragment length polymorphism pattern of the tumor necrosis factor-alpha gene with Ncol. However, they were not significantly different from the control subjects. CONCLUSIONS: Seven restriction fragment length polymorphisms of COL11A2 gene were identified. Two of them (BamHl, 10.0/10.0 kb genotype; HindIII, 19.0/19.0 kb genotype) were significantly different in patients with ossification of the posterior longitudinal ligament.     

Lack of association between the Gly40Ser polymorphism in the glucagon receptor gene and NIDDM in Finland

A heterozygous polymorphism changing GGT40 (Gly) to AGT40 (Ser) (Gly40Ser) in the glucagon receptor gene was reported to be associated with non-insulin-dependent diabetes mellitus (NIDDM). A possible involvement of this polymorphism in impaired glucose tolerance was also suggested in a French population. To replicate this finding we screened 311 unrelated NIDDM patients, 101 unrelated individuals with impaired glucose tolerance and 306 control subjects for the presence of the Gly40Ser polymorphism by use of polymerase chain reaction-restriction fragment length polymorphism in a Finnish population. None of the NIDDM or impaired glucose tolerant patients had this polymorphism. Instead, four of the control subjects (1.3%) were heterozygous carriers of the polymorphism (NS). The age, body mass index, 2-h blood glucose level, 2-h insulin level, and incremental insulin are of the four subjects with this polymorphism were similar to those of the control subjects homozygous for the wild type. Taken together, the data do not support the suggested involvement of the Gly40Ser polymorphism in impaired glucose tolerance and the hypothesis of an association between NIDDM and the glucagon receptor gene in this population.     

Characterization, cell uptake, and subcellular distribution of DNA complexes with lipoglutamides having tetraethylene glycol tails

HLA-G is a non-classical MHC class I gene with a limited tissue distribution. The most pronounced expression is detected in the cytotrophoblast of first trimester placenta. It is possible to detect mRNA for HLA-G in preimplantation blastocysts where expression is correlated with a high cleavage rate of embryos. HLA-G seems to play an important role in the feto-maternal relationship. The polymorphism of the HLA-G locus is not fully clarified. One study has shown extensive nucleotide sequence variation in the exon 3 (alpha-2 domain) in healthy African Americans. A few studies in other populations have only revealed a limited polymorphism. We investigated the polymorphism of the exon 3 of HLA-G by means of Polymerase Chain Reaction (PCR)-Single Strand Conformation Polymorphism (SSCP)- and DNA sequencing analysis in a Danish population. We detected four single-base substitutions in exon 3 compared to the sequence of HLA-6.0 (G*01011); one of these has not been reported before. We also found a deletion of the first base of codon 130 or the third of codon 129 in a heterozygous individual. This study, together with previous results, suggests that the polymorphism of exon 3 of the HLA-G gene in Caucasians is limited, in contrast to that observed in Americans originating from Africa. Implications of this discrepancy and the detected deletion in relation to certain disorders of pregnancy are discussed.     

Nucleotide polymorphisms in the alpha2 gene define multiple alleles that are associated with differences in platelet alpha2 beta1 density

Three allelic differences in the alpha2 gene are associated with expression levels of the alpha2beta1 integrin on the platelet surface. We have previously defined two linked silent polymorphisms in the alpha2 gene coding region at nucleotides 807 (C or T) and 873 (G or A). We have now identified one rarer nucleotide polymorphism in the coding region at nucleotide 837 (T or C) and four additional linked polymorphisms within the introns that flank these coding sequences. Moreover, we have determined that the alloantigenic Br polymorphism, which resides in a distal coding region at nucleotide 1648, is also linked to the 837 polymorphism. Thus, three alpha2 gene alleles, defined by eight nucleotide polymorphisms, have now been discovered. Allele 1 (807T/837T/873A/Brb) is associated with increased levels of alpha2beta1; allele 2 (807C/837T/873G/Brb) and allele 3 (807C/837C/873G/Bra) are each associated with lower levels of alpha2beta1. Finally, we also show here that the rate of platelet attachment to type I collagen in whole blood under conditions of high shear rate (1,500/s) is proportional to the density of alpha2beta1 receptors on the platelet surface. Thus, the density of platelet alpha2beta1 could have an important impact on platelet adhesion to collagen in whole blood and therefore on platelet function in vivo, contributing to an increased risk of thrombosis or to bleeding in relevant disease states.     

Analysis of the p16 gene, CDKN2, in 17 Australian melanoma kindreds

CDKN2 has been implicated as a melanoma susceptibility gene in some kindreds with a family history of this disease. Mutation analysis of CDKN2 in 17 familial melanoma Australian kindreds revealed a paucity of exon mutations and none of the previously described disease-related mutations. One novel germline mutation was found in exon one, Arg24Pro, which segregates with melanoma in 1/17 kindreds. Two previously described polymorphisms, Ala148Thr and a base change at nucleotide 540 were detected and one novel polymorphism in the untranslated region of exon 3 (nucleotide 580) was also found. Together with other recent reports, these findings provide support for CDKN2 as a susceptibility locus for familial melanoma but suggest that other loci are involved in some hereditary melanoma kindreds.     

Reactive arthritis with a severe lesion of the cervical spine

The porphyrias are disorders that result from the inherited or acquired dysregulation of one of the eight enzymes in the heme biosynthetic pathway. Variegate porphyria (VP) is characterized by deficiencies in protoporphyrinogen oxidase (PPO) and has recently been genetically linked (Z = 6.62) to the PPO gene on chromosome 1q21. In this study, we have identified two sequence variants in the PPO gene in a family with VP. The first is a neutral polymorphism at the -47 position of intron 2; this polymorphism is present in the general population and is unlikely to underlie the VP phenotype. The second is a mutation in the PPO gene in a patient with VP; the mutation consists of an apparently de novo 2-bp insertion in exon 3 of PPO and results in a frameshift and downstream premature termination codon. These data establish that a frameshift mutation in PPO is the underlying mutation in this patient with VP and explain the sporadic occurrence of the phenotype in this family.     

Color Doppler ultrasound criteria to diagnose varicoceles: correlation of a new scoring system with physical examination

Mitochondrial aldehyde dehydrogenase (ALDH2) is mainly responsible for the oxidation of acetaldehyde generated during alcohol oxidation in vivo. Cytochrome P-4502E1 (CYP2E1), a liver microsomal enzyme, also metabolizes acetaldehyde and ethanol. Genetic polymorphism of ALDH2 and CYP2E1 was investigated among 481 Korean adults. A new restriction fragment-length polymorphism method was developed to determine the genotype of the ALDH2 alleles. This method proved to be simpler and faster than the hybridization method using allele-specific oligonucleotide probes and polymerase chain reaction-directed mutagenesis. The allele frequencies of ALDH2(1) and ALDH2(2) were 0.840 and 0.160, respectively. This allele frequency of ALDH2(2) is less than in Japanese people. Genetic polymorphism of CYP2E1 was investigated using polymerase chain reaction and restriction fragment-length polymorphism. The estimated allele frequencies for c1 and c2 were 0.808 and 0.192.     

Transverse formamide gradients as a simple and easy way to optimise DNA single-strand conformation polymorphism analysis

Although widely used, the detection of DNA mutations by the single-strand conformation polymorphism (SSCP) method is often hampered by the need to examine a large set of electrophoretic conditions in order to select the one suited to the DNA sequence under study. We show here that the use of transverse chemical gradient gels allows for a quick and easy optimisation of SSCP analysis, as exemplified on two mutations in exon 2 of the alpha-1-antitrypsin gene.     

Polymorphism in alkaline phosphatase (author's transl)

1. In horizontal starch-gel electrophoresis placental extracts show an alkaline phosphatase polymorphism which - according to ROBSON and HARRIS - depends on the fetal genotype. Investigations carried out with a series of 73 placentas revealed 5 of the common 6 phenotypes. Their frequencies were in good accordance with the figures published by the English authors. However, there had been difficulties in typing because of obscrue realizing of the patterns. 2. Although there exists a polymorphism of the heart-stable alkaline phosphatase as observed in 66 serum samples from women at delivery, only four types could be distinguished which, however, could not be related to the placental patterns with any certainty. 3. In all but one of 66 samples of cord serum, investigated simultaneously, a single band only could be observed moving at a slower speed than the serum bands of adult pregnant and non-pregnant persons. This is in conformity with the results of the other investigators. 4. In 18 samples of unheated serum the amount of heat-labile alkaline phosphatase was unimportant and did not alter the features of the heat-stable portion of the enzyme. 5. Contrary to expectations, the alkaline phosphatase polymorphism has no medicolegal relevance.     

Genetic variation in heterogeneous environments

A model of population structure in heterogeneous environments is described and conditions sufficient for maintaining a polymorphism are derived. The absolute fitness of any genotype is regarded as a function of location in the niche space and the population density at that location. Two modes of habitat selection are examined: (1) organisms are distributed uniformly over the environment; and (2) each organism selects to occupy that habitat in which it is most fit ("optimal habitant selection"). Sufficient conditions for maintenance of genetic polymorphisms are derived for both models. In populations which do not practice habitat selection heterozygote superiority averaged over the environment is sufficient to guarantee the existence of polymorphisms. Comparable conditions for populations which practice optimal habitat selction are much less restrictive. If the heterozygotes are superior to one homozygote in any one part of the niche and to the other homozygote in any other part of the niche then a polymorphism will be defined. A positive correlation between genetic and environmental variation follows from the model with habitat selection, but not from the other. The adaptive significance of polymorphisms thus depends on how animals behave.     

Transforming growth factor-beta receptors on human endometrial cells: identification of the type I, II, and III receptors and glycosyl-phosphatidylinositol anchored TGF-beta binding proteins

BACKGROUND: Mutation of the p53 tumor suppressor gene is the most commonly found genetic alteration in human cancer. The E6 gene product of human papillomavirus (HPV) 16 and 18 can inactivate the p53 protein by promoting its degradation. Because most HPV-positive cervical carcinoma cell lines contain wild-type p53 whereas HPV-negative cell lines have point mutations in the p53 gene, a major role in the development of HPV-negative cervical cancer has been attributed to p53. Recent studies, however, have observed no consistent presence of p53 mutation in HPV-negative primary cervical carcinomas. The MDM2 oncogene, which forms an autoregulatory loop with the wild-type p53 protein, has been found amplified in a high percentage of human sarcomas, thus abolishing the antiproliferative function of p53. METHODS: Forty-three primary cervical carcinomas and 10 autopsy-derived distant metastases from one patient were examined for p53 mutation and MDM2 amplification. These tumors had been selected from 238 cervical cancers that had been HPV-typed by Southern blot hybridization and polymerase chain reaction as a representative sample for their HPV status and their clinicopathologic characteristics. Seventeen of the cases had a remarkably good or poor clinical outcome. Human papillomavirus DNA sequences had been detected in 30 of these 43 primary tumors and 13 were negative for HPV by both methods. p53 mutation in the highly conserved exons 5-8 was studied by single-strand conformation polymorphism analysis and direct sequencing. MDM2 amplification was analyzed by Southern blot hybridization. RESULTS: Only two missense point mutations and one nucleotide sequence polymorphism were detected: a TAC-->TGC transition in codon 234 in exon 7, resulting in a Tyr-->Lys substitution, a CGT-->TGT transition in codon 273 in exon 8, resulting in an Arg-->Cys substitution and a polymorphism (CGA-->CGG) in codon 213 in exon 6. Both tumors revealing the point mutations were HPV-negative carcinomas. Amplification of the MDM2 gene was observed in 1 of the 53 specimens tested. CONCLUSIONS: In contrast to data derived from cultured cervical carcinoma cell lines and primary sarcomas, these results indicate that p53 mutation and amplification of the MDM2 oncogene are rare even in HPV-negative primary cervical carcinomas. However, to the authors; knowledge, this is the first observation of MDM2 amplification in humans outside sarcomas and neuroepithelial tumors.     

Pharmacogenetics: a laboratory tool for optimizing therapeutic efficiency

Pharmacogenetics is the study of the linkage between an individual's genotype and that individual's ability to metabolize a foreign compound. Differences in metabolism of therapeutics can lead to severe toxicity or therapeutic failure by altering the relation between dose and blood concentration of the pharmacologically active drug. Phenotypes exhibiting poor and ultraextensive metabolism result from genetic variance (polymorphism) of enzymes involved in metabolism. Thus, in pharmacogenetic studies one applies genotyping of polymorphic alleles encoding drug-metabolizing enzymes to the identification of an individual's drug metabolism phenotype. This knowledge, when applied to dosing or drug selection, can avoid adverse reactions or therapeutic failure and thus enhance therapeutic efficiency. More than 25 commonly prescribed medicines are metabolized by the cytochrome P-4502D6 (CYP2D6) isoenzyme, and polymorphism of the CYP2D6 gene affects the therapeutic management of up to 17% of individuals in some ethnic groups. In this review, we summarize and update information concerning drug-metabolizing genotypes with emphasis on CYP2D6 genotyping techniques that can be applied by the clinical laboratory for linking human genetics to therapeutic management.