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At least 955 records · Page 53Linked to original sources
PubMed1997

Amino-acid substitution in alpha-spectrin commonly coinherited with nondominant hereditary spherocytosis.

Nondominant hereditary spherocytosis (ndHS) is a disorder characterized in some patients by severe hemolytic anemia and marked deficiency of erythrocyte spectrin. This report describes the identification of a variant spectrin chain, alpha-spectrin Bughill or alpha(BH), that is associated with this disorder in a number of patients. Tryptic maps of spectrin from affected individuals revealed an acidic shift in isoelectric point of the alphaII domain peptides at 46 kD and 35 kD. A point mutation at codon 970 of the alpha-spectrin gene (GCT-->GAT), that changes the encoded amino acid from an alanine to an aspartic acid, was identified in genomic DNA of affected patients. The alpha(BH) variant was present in 8 patients with ndHS from five different kindreds but was absent in 4 patients from two other kindreds. The 8 ndHS patients with the alpha(BH) variant appeared to be homozygous for the alpha(BH) variant by analysis of peptide maps of limited tryptic digests of erythrocyte spectrin. However, following genomic DNA analysis, only 2 of these patients were true homozygotes, whereas 6 were found to be doubly heterozygous for the alpha(BH) allele and a second, presumably abnormal, alpha-spectrin gene. These results suggest that, in these 6 patients, the second alpha-spectrin allele is in fact associated with one or more genetic defect(s), causing decreased accumulation of alpha-spectrin. The pattern of transmission of the alpha(BH) allele in certain families suggests that the alpha(BH) amino-acid substitution is not itself responsible for ndHS but is more likely a polymorphic variant that, in some but not all cases, is in linkage disequilibrium with another uncharacterized alpha-spectrin gene defect that itself is a cause of ndHS.

Alleles↗
PubMed1999

Sensitive median-ulnar motor comparative techniques in carpal tunnel syndrome.

We describe two modified methods for median-to-ulnar motor conduction comparison in the diagnosis of median neuropathy at the wrist: the median-thenar to ulnar-thenar latency difference (TTLD), and the median-thenar to ulnar-hypothenar latency difference (THLD). We also describe an F-wave ulnar-to-median comparative test, the F-wave latency difference (FWLD). The abnormal cutoffs based upon 34 normal controls are: TTLD, 0.8 ms; THLD, 1.2 ms; FWLD, 0.6 ms. In 50 patients (79 hands) with clinically defined carpal tunnel syndrome and electrophysiological evidence of median neuropathy at the wrist (based upon a prolonged median nerve palm-wrist latency), the diagnostic sensitivities were: 95-98%, 85-88%, and 75-78%, respectively. These tests are therefore highly sensitive. They are easily performed and require minimal additional effort to incorporate into commonly used clinical electrodiagnostic routines. They may be advantageous when a concomitant polyneuropathy is present, and they may also help avoid technical pitfalls and aid in identification of anatomic variants.

Action Potentials↗
PubMed2000

Alpha1-antitrypsin in serum determined by capillary isoelectric focusing.

A capillary isoelectric focusing (CIEF) method using bare fused-silica capillaries filled with polyethylene oxide (PEO) and carrier ampholyte solutions in the pH 3.5-5.0 range has been developed for the identification of alpha1-antitrypsin (alpha1AT) phenotypes in human serum. This novel procedure was routinely applied to the study of serum samples of five controls whose alpha1AT phenotype was previously identified and of twelve subjects whose alpha1AT phenotype was unknown. The results obtained allowed us to confirm or identify the alpha1AT phenotype in all sera tested. This procedure seems particularly suitable for identification of alpha1AT variants associated with diseases of clinical relevance.

Chromatography, High Pressure Liquid↗
PubMed1977

Identification of hemoglobins in single erythrocytes by electrophoresis.

We present here a method for electrophoretic identification of hemoglobin variants in many individual erythrocytes simultaneously, which is made possible by instruments developed in our laboratory. The system spaces and aligns the erythrocytes along a common origin and prevents 'doubling' of cells. Unambiguous separation of different hemoglobins (Hbs) occurred from single erythrocytes, such as Hbs A, C, F. Staged micrographs showed that hemoglobins of differing mobility from a single cell arise from a single point source of hemoglobin. In principle this capability is applicable to other proteins and could facilitate investigation at a cellular level into a variety of questions pertaining to gene action.

Electrophoresis↗
PubMed1988

Hemoglobin Pasadena: identification of the gene mutant by DNA analysis using synthetic DNA probes.

Hemoglobin Pasadena [beta 75(E19)Leu----Arg] was found in a boy who had an acute episode of anemia and rapid splenic enlargement. His father was the only other member of a large family with this hemoglobinopathy. We have used gene mapping techniques for direct identification of the beta-globin gene mutation. To correlate the DNA findings with the structural identification of this variant, we have also performed globin chain separation and analysis of the tryptic peptides using high performance liquid chromatography and secondary ion mass spectral analysis.

Adult↗
PubMed2005

Pleomorphic lobular carcinoma of the breast: role of comprehensive molecular pathology in characterization of an entity.

Immunohistochemical analysis of E-cadherin has changed the way lobular neoplasia is perceived. It has helped to classify difficult cases of carcinoma in situ with indeterminate features and led to the identification of new variants of lobular carcinoma. Pleomorphic lobular carcinoma (PLC) and pleomorphic lobular carcinoma in situ (PLCIS), recently described variants of invasive and in situ classic lobular carcinoma, are reported to be associated with more aggressive clinical behaviour. Although PLC/PLCIS show morphological features of classic lobular neoplasia and lack E-cadherin expression, it is still unclear whether these lesions evolve through the same genetic pathway as lobular carcinomas or are high-grade ductal neoplasms that have lost E-cadherin. Here we have analysed a case of extensive PLCIS and invasive PLC associated with areas of E-cadherin-negative carcinoma in situ with indeterminate features, using immunohistochemistry, chromogenic in situ hybridization, high-resolution comparative genomic hybridization (CGH) and array-based CGH. We observed that all lesions lacked E-cadherin and beta-catenin and showed gain of 1q and loss of 16q, features that are typical of lobular carcinomas but are not seen in high-grade ductal lesions. In addition, amplifications of c-myc and HER2 were detected in the pleomorphic components, which may account for the high-grade features in this case and the reported aggressive clinical behaviour of these lesions. Taken together, these data suggest that at least some PLCs may evolve from the same precursor or through the same genetic pathway as classic lobular carcinomas.

Adult↗
PubMed2002

Toxicology of a Microcystis ichthyoblabe waterbloom from Lake Oued Mellah (Morocco).

In the Lake Oued Mellah cyanobacteria waterblooms occur periodically in late spring and summer with Microcystis ichthyoblabe as the main bloom-forming species. In 1999, a heavy waterbloom of M. ichthyoblabe occurred during May June with a maximal biomass of 298 mg/l. During this period, several bloom samples were collected. The toxicity assessment was done by mouse and brine shrimp (Artemia) bioassays. Apart from the sample collected on 15/06/1999, all the other samples were toxic by mouse bioassay. The LD50 determined by intraperitoneal injection to mice during active cyanobacterial growth and decline phases were 518 and 1924 mgDW/kg respectively. Using Artemia bioassay, the 24hLC50 varied from 6.0 to 40.6 mg/ml and the 48hLC50 ranged from 2.8 to 18.2 mg/ml. The separation and identification of microcystin variants was performed by high performance liquid chromatography-photodiode array detection. Eleven toxins were separated and preliminarily identified as microcystin variants as they exhibit a typical UV spectra like the microcystin-LR standard. The quantification of total microcystins determined by enzyme-linked immunosorbent assay showed that the contents were varied between 0.1 and 0.76 microgram/g DW.

Animals↗
PubMed2002

Detection and quantification of microcystins from cyanobacteria strains isolated from reservoirs and ponds in Morocco.

In Morocco, the occurrence of toxic cyanobacteria blooms is confirmed in some water bodies used for recreational and/or as drinking water reservoirs. According to WHO recommendations, the establishment of a monitoring program for microcystins is a necessity. This paper presents toxicological studies of 19 toxic cyanobacteria strains of Microcystis, Synechocystis, Pseudanabaena, and Oscillatoria. These strains were isolated from various water bodies including natural lakes, reservoirs, and ponds located in central regions of Morocco. The isolation, culture, and biomass production of these strains was made on Z8 or BG13 media under laboratory controlled conditions. The hepatotoxicity of cyanobacterial lyophilized material was confirmed by mouse bioassays. The amount of microcystins produced by each strain was determined by the enzyme-linked immunosorbent assay (ELISA). The detection and identification of microcystin variants was performed by high performance liquid chromatography (HPLC) with photodiode array detection. Almost all strains showed medium to high toxicity, the estimated LD50 i.p. mice bioassay ranged between 28 to 350 mg/kg body weight. The concentrations of microcystins varied between 2.16 to 944 micrograms/g and 26.8 to 1884 micrograms/g dry weight determined by ELISA and HPLC, respectively. The screening of bloom-forming and microcystin producer cyanobacteria strains in these fresh water bodies leads us to propose the need for the establishment of a survey of cyanobacteria and a cyanotoxin-monitoring program.

Animals↗
PubMed1995

PreS and core gene heterogeneity in hepatitis B virus (HBV) genomes isolated from patients with long-lasting HBV chronic infection.

Several reports have been recently published concerning the identification of HBV variants due to rearrangements of the preS1/preS2 or core regions of the viral genome. To evaluate the frequency of the natural occurrence of such variants and whether the heterogeneity of these genomic regions correlates with the severity of the liver disease, we have examined the preS1/preS2 region and the entire core gene sequences of HBV DNA isolated from sera of 30 chronic HBV carriers, 7 with chronic persistent hepatitis, 10 with chronic active hepatitis, 7 with cirrhosis, and 6 with hepatocellular carcinoma. We found no significant rearrangement in any of the preS1 regions examined, while genomic modifications precluding the preS2 protein production were detected in 4 cases, 2 with cirrhosis and 2 with hepatocellular carcinoma. The analysis of the core gene showed the presence of various numbers of missense mutations in the core region of most cases, independent of the grade of liver disease. Moreover, in contrast with previous reports, neither mutation cluster region nor deletion was observed. On the contrary, HBV strains with the precore mutation at nucleotide position 1896, effecting the rise of HBeAg-defective viruses, were found in 26 of the 30 cases examined. In conclusion, our data show that the precore mutant is the only HBV genomic variant commonly selected during a chronic infection. Other HBV variants, due to genomic rearrangements outside the precore region, may exist and influence the outcome of the infection and the course of the liver disease, but the emergence of each of these variants seems to be an unusual and probably casual event.

Adolescent↗
PubMed2000

Identification of functional differences between prototype Epstein-Barr virus-encoded LMP1 and a nasopharyngeal carcinoma-derived LMP1 in human epithelial cells.

The contribution of Epstein-Barr virus (EBV) strain variation to the pathogenesis of virus-associated tumours remains unknown. Given the central role of LMP1 in EBV-induced transformation, much interest has focused on the influence of LMP1 sequence variation on the signaling pathways and multiple downstream phenotypic consequences of LMP1 expression. The identification of LMP1 variants with a common 10-amino-acid deletion and additional point mutations (typified by the CAO-LMP1 isolate) in EBV strains associated with nasopharyngeal carcinoma prompted us to examine the effect of stable prototype B95.8-LMP1 and CAO-LMP1 expression on the phenotype and differentiation of SCC12F human epithelial cells. Both forms of LMP1 were able to induce expression of the antiapoptotic A20 protein and provide protection from tumour necrosis factor-alpha-induced cytotoxicity. Although B95.8-LMP1 induced growth inhibition, expression of certain cell surface molecules (CD40, CD44, and CD54), and secretion of interleukin-6 and -8 in SCC12F cells, stable CAO-LMP1 expression failed to elicit these effects. Furthermore, B95. 8-LMP1, but not CAO-LMP1, induced alterations in cell morphology and blocked epithelial cell differentiation. Both B95.8-LMP1 and CAO-LMP1 induced similar levels of nuclear factor-kappaB activation, but the ability of CAO-LMP1 to activate the AP-1 pathway was relatively impaired. These data highlight significant functional differences between the prototype B95.8-LMP1 and the CAO-LMP1 variant when stably expressed in human epithelial cells and suggest that continued analysis of LMP1 variants will help to further dissect the signaling pathways activated by LMP1 as well as provide insights into the contribution of LMP1 sequence variation to the pathogenesis of EBV-associated tumours.

Antigens, CD↗
PubMed1994

Molecular characterization of thirteen gyrA mutations conferring nalidixic acid resistance in Bacillus subtilis.

We isolated 607 independent nalidixic acid-resistant mutants from Bacillus subtilis. A 163 bp DNA segment from a 5' portion of the gyrA gene was amplified from the DNA of each mutant strain. After heat denaturation, the product was subjected to gel electrophoresis to detect conformational polymorphism of single-strand DNA (PCR-SSCP analysis). Mobility patterns of the two DNA strands from all the mutant strains examined differed from those of the parental wild-type strains. The patterns were classified into 13 types, and the DNA sequence of each type was determined. A unique sequence alteration was found in mutants belonging to each of the 13 types, defining 13 gyrA alleles. Eight were single base pair substitutions, four were substitutions of two consecutive base pairs and one was a substitution of three consecutive base pairs. Only three amino acid residues (Ser-84, Ala-85, and Glu-88) were altered in the deduced amino acid sequences of the mutated genes. We conclude that molecular typing based on the PCR-SSCP method is a powerful technique for the exhaustive identification of allelic variants among mutants selected for a phenotypic trait.

Bacillus subtilis↗
PubMed1978

Electrophoretic heterogeneity of 5-phosphoribosyl-1-pyrophosphate synthetase within and among humans.

The product of the enzyme 5-phosphoribosyl-1-pyrophosphate (PPriboseP) synthetase is a substrate for purine, pyrimidine, and pyridine biosynthesis and may be rate limiting for purine biosynthesis. A system developed to electrophoretically separate and histochemically detect PPriboseP synthetase in crude cell extracts has facilitated the identification of electrophoretically variant enzyme forms in the erythrocytes of five patients from a patient population of 200. Additional studies of human organs obtained at autopsy revealed a unique electrophoretic mobility for nearly each organ within the same individual.

Electrophoresis, Cellulose Acetate↗
PubMed2005

[The value of pharmacogenetic tests in antidepressive medication therapy].

The pharmacokinetics and effect of antidepressants are influenced by genetic factors. Modern methods of genotyping allow fast and inexpensive identification of genetic variants and thus can be used in clinical diagnostics to improve the tolerance to drug therapy. Numerous studies have investigated the significance of genetic variants in drug-metabolizing enzymes, drug and natural substrate transporters, neurotransmitter receptors, and molecules involved in signal transduction. While the interindividual differences in oral clearance, half-life, and bioavailability caused by genetic variants in the cytochrome P450 liver enzymes can be overcome by individual adjustment of dosage according to certain genotypes, the effects of genetic variants in antidepressive target structures are more difficult to translate into clinical recommendations. This article gives an overview of the currently available literature and points to situations in which the determination of pharmacogenetic variants might change drug therapy or therapeutic strategies for the individual patient. Dose adjustments for common antidepressant drugs based upon differences in pharmacokinetic parameters caused by genetic variability will be given.

Antidepressive Agents↗
PubMed2005

Molecular characterization of new waxy mutants identified in bread and durum wheat.

Recently, electrophoretic analyses of waxy proteins in several hexaploid and tetraploid wheat accessions from worldwide collections have permitted the identification of new variants at the waxy loci, including allelic forms with different mobilities and partial null types. In this paper, the molecular characterization of mutated waxy loci in four bread wheat cultivars (two lacking the Wx-B1 and two lacking the Wx-D1 protein, respectively) and in four durum wheat cultivars (one lacking Wx-A1 and the remainder with Wx-B1 proteins showing different electrophoretic mobilities) was conducted by means of PCR, Southern and DNA sequence analyses. Three primer pairs were developed that identified six of the above-mentioned mutations and allowed their molecular description, providing a useful tool for further germplasm screening or marker assisted progeny selection in breeding programs involving the newly identified material. We have found that a complete gene deletion is responsible for a null allele at the Wx-B1 locus in one bread wheat line, whereas sequencing of the corresponding fragments showed a 724 bp deletion in the Wx-D1 locus in one line of bread wheat and an insertion of 89 bp in the Wx-A1 locus in one line of durum wheat, respectively. In addition, nucleotide substitutions and various insertions/deletions ranging from 3 to 30 bp were detected in the PCR fragments of one durum wheat line with a Wx-B1 protein with a different electrophoretic mobility. A fourth primer set, specific for this mutation, was consequently derived.

Base Sequence↗
PubMed2006

hTERT immunopositivity patterns in the normal brain and in astrocytic tumors.

Accumulating data about the impact of hTERT in astrocytic tumor carcinogenesis and recent evidence about its association with disease outcome prompt the evaluation of this molecule with methods applicable in routine pathology practice. In this study, we investigated hTERT protein expression with immunohistochemistry (IHC) and the NCL-hTERT antibody in 49 astrocytic tumors. Results were validated with the assessment of hTERT mRNA (relative quantification, identification of splice variants, in situ hybridization). Specific nuclear hTERT immunostaining patterns (IPs) were characterized as patterns As (single large dot) and Am (multiple dots) without nucleoplasm staining and pattern B (nucleoplasm staining with or without dots), corresponding to low and high relative hTERT expression values (P<0.0001). Low- and high-grade astrocytic tumors were found positive for hTERT in 74 and 85% of cases, respectively. Heterogeneity in the distribution of hTERT-positive cells was observed in all tumors. The prevailing nuclear IPs differed significantly between pilocytic astrocytomas (pattern As) and the rest of histologic types up to glioblastoma (patterns Am and B) (P<0.0001). The described nuclear IPs were also observed in non-neoplastic cells. Positive endothelial cells were found in astrocytic tumors of all grades, even when tumor cells showed no hTERT immunoreactivity. A subset of mature normal neurons was positive for hTERT (pattern As), suggesting a role for this molecule in neuronal maintenance in the adult brain. The nuclear hTERT IPs described here may reflect the functional status of non-neoplastic brain and neoplastic astrocytic cells and support the model of a continuum in the development of glioblastomas from diffuse fibrillary astrocytomas.

Adult↗
PubMed1999

Hyperbilirubinemia, glucose-6-phosphate-dehydrogenase deficiency and Gilbert's syndrome.

The pathogenesis of neonatal hyperbilirubinemia has not yet been completely defined in normal and glucose-6-phosphate-dehydrogenase (G6PD)-deficient newborns. The recent identification of a variant promoter in the gene encoding for the bilirubin uridine-diphosphoglucuronosyl-transferase (UGT-1 A) associated with Gilbert's syndrome, allowed us to explore whether the presence of this variant promoter is a risk factor for the development of neonatal hyperbilirubinemia in normal newborns and in association with G6PD deficiency. We found that the variant (TA)7/(TA)7 promoter shows no statistically significant difference in normal or G6PD-deficient newborns developing severe hyperbilirubinemia and in control subjects from the same population. This finding indicates that the variant promoter of UGT-1 A does not contribute to the development of hyperbilirubinemia in the newborn.

Adult↗
PubMed2006

A cluster analysis to define human aging phenotypes.

The definition of a precise and consistent aging phenotype that allows to measure the physical and cognitive decline, as well as the increase of mortality hazard late in life, is a major problem for studies aimed at finding the genetic factors modulating rate and quality of human aging. In this frame, it seems promising the concept of frailty which tends to figure out the subjects who are more vulnerable and more prone to negative outcomes, such as death or hospitalization. Cognitive, functional and psychological measures turned out to be the most effective measures to define frailty, as they condense most of the frailty cycle that occurs in the elderly and is probably responsible of the aging related physical decline. We used MMSE, Hand Grip strength, and GDS as variable parameters in a hierarchical Cluster Analysis (CA) in order to recognise aging phenotypes. By using a sample of 65-85 years old subjects we identified three frailty phenotypes that were consistent from both geriatric and genetic perspectives. Therefore, the method we propose may provide unbiased phenotypes suitable for the identification of genetic variants affecting the quality of aging in this age range. The CA method was less effective in ultranonagenarians, probably due to the high prevalence of frail subjects in this age group that makes difficult to distinguish discrete phenotypes.

Aged↗
PubMed2000

Update on inflammatory bowel disease genetics.

The idiopathic inflammatory bowel diseases (IBD), comprised of Crohn's disease (CD) and ulcerative colitis (UC), are related, complex genetic disorders. With the completion of the human genomic sequence, identification of genetic variants contributing to IBD susceptibility can now more systematically be identified. Significant genetic linkages have been observed on chromosomes 16, 12, 14, 19, 6, and 1, of which the linkage to CD on chromosome 16 is the most well-established. For many of the other regions, evidence for linkage has been observed for both CD and UC. Candidate gene association studies have largely focused on genes involved in inflammatory pathways, such as cytokines and cytokine receptors. With greater understanding of genetic differences underlying both disease susceptibility and response to medical therapy, the individualization of medical approaches based on this knowledge may soon be possible in patients with IBD.

Base Sequence↗