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Variants of hamster fibroblasts resistant to Ricinus communis toxin (ricin).

1. Variant baby-hamster kidney (BHK) cell lines were isolated that grow in the presence of high concentrations of ricin, the toxic lectin of castor beans (Ricinus communis). The variant lines were independently derived from several cultures of normal BHK cells which had been exposed to the mutagen, methyl-N-nitro-N-nitrosoguanidine, before selection by ricin. 2. The cell lines maintain a high degree of resistance to ricin after growth in lectin-free medium for prolonged periods and therefore exhibit stable phenotypes that are different from normal BHK cells. 3. A preliminary classification of the phenotypes was made. Several cell lines bind normal amounts of 125I-labelled ricin, whereas other bind the lectin poorly. 4. A loss of surface receptors for two other lectins, R. communis RCA and Axinella polyploides, which have specificities similar to ricin, was also found in some but not all of the cell lines showing decreased surface concentrations of ricin receptors. 5. The binding to the ricin-resistant cells of lectins of different sugar specificity, namely Lens culinaris lectin and concanavalin A, was similar to, or higher than, to normal BHK cells. 6. Several of the ricin-resistant cell lines were shown to be cross-resistant to the weak cytotoxicity of Phaseolus vulgaris lectin. By contrast, some cell lines were more sensitive to concanavalin A than were normal BHK cells.

Agglutination↗

Diverse point mutations in the human gene for polymorphic N-acetyltransferase.

Classification of humans as rapid or slow acetylators is based on hereditary differences in rates of N-acetylation of therapeutic and carcinogenic agents, but N-acetylation of certain arylamine drugs displays no genetic variations. Two highly homologous human genes for N-acetyltransferase (NAT; arylamine acetyltransferase, acetyl CoA:arylamine N-acetyltransferase, EC 2.3.1.5), NAT1 and NAT2, presumably code for the genetically invariant and variant NAT proteins, respectively. In the present investigation, 1.9-kilobase human genomic EcoRI fragments encoding NAT2 were generated by the polymerase chain reaction with liver and leukocyte DNA from seven subjects phenotyped as homozygous and heterozygous acetylators. Direct sequencing revealed multiple point mutations in the coding region of two distinct NAT2 variants. One of these was derived from leukocytes of a slow acetylator and was distinguished by a silent mutation (codon 94) and a separate G----A transition (position 590) leading to replacement of Arg-197 by Gln; the mutated guanine was part of a CpG dinucleotide and a Taq I site. The second NAT2 variant originated from liver with low N-acetylation activity. It was characterized by three nucleotide transitions giving rise to a silent mutation (codon 161), accompanied by obliteration of the sole Kpn I site, and two amino acid substitutions: Thr for Ile (codon 114) and Arg for Lys (codon 268). Heterozygosity was detected in three NAT2 samples: two were heterozygous for the rapid and one of the allelic variants, and the third was a compound heterozygote of both mutant alleles. The results show conclusively that the genetically variant NAT is encoded by NAT2.

Acetylation↗

Comparative analyses of members of the Venezuelan equine encephalomyelitis virus complex.

Polyacrylamide gel electrophoretic examination of viruses selected from the Venezuelan equine encephalomyelitis (VEE) complex revealed distinct strain to strain differences in profiles of the two virion envelope proteins. The core protein was identical in all viruses tested. We detected five electrophoretic patterns into which the virus strains could be classified and these were designated alpha (alpha), beta (beta), gamma (gamma), delta (delta), and episolon (episolon). Isolates representing variant E of subtype I exhibited a profile characterized by only one apparent envelope band. The epizootic subtypes I-A, I-B, I-C and the sylvatic subtype II viruses contained at least two envelope proteins which differed in molecular weight according to virus strain but which were not necessarily specific for antigenic variety. These results generally, though not uniformly, support the serologic classification of the VEE virus complex and suggested that the usefulness of the classification scheme could be complemented by the inclusion of biochemical criteria.

Animals↗

Expression and prognostic value of CD44 standard and variant v3 and v6 isoforms in prostate cancer.

BACKGROUND: The adhesion molecule CD44 standard (CD44s), and its variant isoforms v3 and v6 are associated with cell-to-cell adhesion. The down-regulation of CD44 standard and its variant isoform CD44v6 is linked with early cancer cell dissemination, but the relationship between CD44v3 and malignant features of prostate cancer (PC) has not been established previously. METHODS: The expression of CD44s and its CD44v3 and CD44v6 isoforms was analysed by immunohistochemistry in 209 archival PC biopsy specimens to establish their prognostic value. RESULTS: Down-regulation of CD44s and CD44v6 was related to high T classification, metastasis, high Gleason score, DNA aneuploidy, high S-phase fraction, high mitotic index, perineural growth and dense amount of tumour infiltrating lymphocytes (p < 0.03 for all). Down-regulation of CD44s and CD44v6 was related to poor survival in the entire cohort (p < 0.0001), in M0 tumours (p < 0.001) and in T1-2M0 tumours (p < 0.05). In needle biopsies and TURP specimens, the prognostic impact of the investigated parameters was similar. In the multivariate analysis, T classification (p = 0.0009), presence of metastases (p < 0.0001), Gleason score (p = 0.0060) and CD44v6 (p = 0.0220) expression were independent prognostic factors. In M0 tumours, T classification (p < 0.0001) and CD44v6 (p = 0.003) independently predicted survival. CONCLUSION: Down-regulation of CD44s and its CD44v6 isoform is related to tumour malignancy and unfavourable prognosis in PC.

Aged↗

Characterization of a relatively rare class B, type 2 trypanosome variant surface glycoprotein gene.

The variant surface glycoprotein (VSG) gene of Trypanosoma brucei rhodesiense LouTat 1.5, a defined African trypanosome variant antigenic type, was cloned and sequenced. Southern blot analysis revealed 2 DNA restriction fragments in both VSG 1.5 expressor and nonexpressor populations, suggesting that there are 2 genomic copies of the VSG 1.5 gene and no expression-linked copies. Pulsed-field gel electrophoresis followed by Southern blot analysis showed that each copy of the VSG 1.5 gene exists on a separate large chromosome in both the expressor (approximately 3.5- and 4-megabase (Mb) chromosomes) and nonexpressor (approximately 4- and 5.7-Mb chromosomes) populations. Thus, VSG genes may be present on larger chromosomes than previously reported. Sequence analysis and alignments revealed that the VSG 1.5 molecule is a class B VSG with 12 cysteine residues in the N-terminus and is classified as a type 2 VSG based on C-terminus motifs. This classification shows that the VSG 1.5 molecule represents a relatively rare VSG class and type. Taken together, these studies provide additional information on VSG genes and proteins and supply the foundation for structure-function analysis of the VSG 1.5 surface antigen expressed by trypanosomes of the LouTat 1 serodeme.

Amino Acid Sequence↗

The association of genetic polymorphisms in sex hormone biosynthesis and action with insulin sensitivity and diabetes mellitus in women at midlife.

We evaluated associations of insulin sensitivity, metabolic syndrome, and diabetes mellitus with single nucleotide polymorphism (SNP) variants from sex hormone biosynthesis and action in women of 4 races/ethnicities. DNA was extracted from transformed cell samples of 1,538 women in the Study of Women's Health Across the Nation (SWAN). African American, Caucasian, Chinese, and Japanese women in SWAN enrolled in the Sex Steroid Hormone Genetics Protocol from whom fasting glucose and insulin measures (for estimating insulin sensitivity), diabetes status, and metabolic syndrome classification were obtained. SNPs from the genes encoding aromatase (CYP 19), 17beta-hydroxysteroid dehydrogenase (17HSD) type 1, and the estrogen receptors-alpha (ESR1) and -beta (ESR2) were measured. The prevalence of metabolic syndrome was 20% in Chinese women, 22% in Japanese women, 28% in Caucasian women, and 43% in African American women. The prevalence of diabetes was 3% in Chinese women, 4% in Japanese women, 7% in Caucasian women, and 19% in African American women. Significant associations of CYP 19 genotypes and insulin sensitivity were observed in African American, Caucasian, and Japanese women. Selected ESR1 and ESR2 genotypes were associated with insulin sensitivity and metabolic syndrome only in Japanese and Chinese women. The strongest associations related 17HSD genotypes to diabetes in Caucasian women, with odds ratios ranging from 4.4 to 7.5 and confidence intervals that excluded the null value. We observed strong associations between variations in sex hormone biosynthesis and function genes with insulin sensitivity, the metabolic syndrome, and diabetes that varied by race/ethnicity. The strong association of 17HSD and diabetes in Caucasian women has not been previously reported and should be further investigated.

17-Hydroxysteroid Dehydrogenases↗

Expression of E-cadherin, and CD44s and CD44v6 and its association with prognosis in head and neck cancer.

OBJECTIVES: In the current study, the expression of E-cadherin, CD44s, and CD44v6 has been noted as markers for tumor metastasis and prognosis in several tumors, so we examined whether or not E-cadherin, CD44s, and CD44v6 are useful markers for evaluating the prognosis of mesopharyngeal cancer patients. METHODS: The expression of E-cadherin, CD44s, and CD44v6, was evaluated immunohistochemically using monoclonal antibodies against epitopes of standard and variant proteins, in paraffin-embedded mesopharyngeal cancer tissues from 57 patients who had received curative therapy. RESULTS: Tumor tissues from 47 (82.5%) patients showed positive immunoreactivity with monoclonal antibody against E-cadherin, 43 (75.4%) patients showed positive expression with CD44, and 45 (78.9%) patients showed positive expression with CD44v6. The expression of CD44v6 was slightly correlated with tumor volume, and lymph node metastasis, and stage classification (P > 0.05). However, there was no significant correlation between the expression of E-cadherin, CD44s and CD44v6 and clinicopathological characteristics. Concerning the prognosis, the survival period of patients with CD44s positive tumors was shorter than that of patients with CD44s negative tumors (18.2% versus 52.1%, 5-year survival, P > 0.05). The survival period of patients with CD44v6 positive tumors was also shorter than that of patients with CD44v6 negative tumors (12.8% versus 55.6%, 5-year survival, P > 0.05). CONCLUSION: These results suggest that CD44v6 may be related to tumor invasion and metastasis, and both CD44s and CD44v6 may be useful markers for poor prognosis in head and neck cancer.

Adult↗

HLA-B27 genotyping by fluorescent resonance emission transfer (FRET) probes in real-time PCR.

Several polymerase chain reaction (PCR)-based human leukocyte antigen (HLA) genotyping methods are in use, but none is fully satisfactory. The introduction of real-time PCR (rt-PCR) with fluorescence resonance energy transfer (FRET) probes provides a powerful tool to overcome the drawbacks of current methods such as the long processing time and the requirement for post-PCR manual procedures. Here we present evidence that the FRET-fluorotyping principle may resolve HLA-B27 variants, providing a higher resolution in less time than the techniques currently in use. The protocol uses between one and three consecutive amplification reactions depending on the resolution required. The first reaction, aimed at detecting HLA-B27-positive samples, uses beta-globin coamplification as control. The second reaction, aimed at resolving most frequent B27 alleles, uses two hybridization probes whose melting temperatures curves allow the classification of HLA-B27 alleles into eight groups. By adding a third reaction, even the rarest alleles associated and not associated to ankylosing spondylitis (AS) may be discriminated. The technique was blindly tested on 60 samples from individuals previously typed and confirmed by standard PCR sequence-specific oligoprobes-PCR sequence and PCR-based typing PCR-SBT (30 B27+, 30 non-B27). There was a complete concordance rate, thus confirming the potential of this new technique for clinical HLA-B27 typing and for HLA typing in general.

Cell Line, Transformed↗

Anorexia nervosa with excessive exercise: a phenotype with close links to obsessive-compulsive disorder.

Anorexia nervosa (AN) and obsessive-compulsive disorder (OCD) are highly comorbid, and appear to share a common neurophysiological dysfunction that contributes to the obsessional thoughts and compulsive behaviours seen in both disorders. Obsessive-compulsive personality (OCP) traits are also important risk factors for AN. Since excessive exercise has also been associated with greater obsessionality, we hypothesised that AN patients with a hyperactive behavioural profile represent a phenotype more closely linked to OCD than their non-exercising counterparts. We examined prospectively 50 female AN-Restrictor patients whom we classified as "excessive" or "non-excessive" based on their exercise status i) at admission and ii) over the lifetime of their illness. Validated measures of OCD symptoms and OCP traits were obtained at admission and after refeeding at discharge. On both classification methods, excessive exercisers had greater OCD symptoms and OCP traits than the non-excessive group, but did not differ on body mass index. OCD symptoms, but not OCP traits, decreased between admission and discharge. Findings support our prediction that AN patients with excessive physical activity constitute a subtype of the disorder with strong links to OCD. Indeed, this phenotype may be a culture-bound variant of OCD.

Adolescent↗

Familial medial telangiectatic nevus: variant of nevus flammeus--port-wine stain.

Six families in which a few members, in three generations, were affected with medial telangiectatic nevus (salmon patch, stork bite, angel's kiss) on the forehead, glabella, upper eyelids, upper lip, nose, and nuchal and occipital areas are presented. This is a mild variant of lateral telangiectatic nevus (nevus flammeus, port-wine stain) that disappears in about 50 percent of patients during the first years of life. In one family, lateral telangiectatic nevus (nevus flammeus, port-wine stain) and superficial (strawberry) hemangioma coexisted with medial telangiectatic nevus. This paper discusses the familial incidence of medial telangiectatic nevus and a new modality of treatment. Moreover, the paper presents a classification of vascular malformations and proposes a new terminology.

Adult↗

Extraskeletal myxoid chondrosarcoma: updated clinicopathological and molecular genetic characteristics.

Extraskeletal myxoid chondrosarcoma (EMC) is a rare soft-tissue sarcoma characterized by distinctive morphological and cytogenetical features. As its name implies, EMC was believed to represent a variant of soft-tissue chondrosarcoma owing to its histological resemblance to chondroblastic tissue in the early stages of cartilage development or chondroid tumors such as skeletal chondrosarcoma. However, the chondroid nature has been a subject of controversy, and its line of differentiation remains to be determined. Consequently, the tumor is provisionally classified into a group of tumors of uncertain differentiation in the revised World Health Organization classification of tumors of soft tissue and bone. Moreover, immunohistochemical and ultrastructural features of neural or neuroendocrine differentiation have been recently reported in a subset of EMC, providing a new insight into their histogenetic nature. Chromosomal rearrangements involving 9q22, such as t(9;22)(q22;q12), and resultant NR4A3 fusion genes are tumor-type specific or pathognomotic for this entity and are assumed to play an important role in the development of EMC. Although the biological mechanisms and functions are largely unknown, the NR4A3-related pathway is considered a potential molecular target for future therapeutic intervention. Because of its protracted but resilient nature, a tenacious and long-term follow up is necessary for any patient.

Chondrosarcoma↗

[MRI of the brain in the evaluation of children with developmental delay].

PURPOSE: To analyze the diagnostic value of MRI in children with developmental delay. Materials and Methods. From 1991 to 1997, 224 examinations were performed. Retrospective analysis of clinical findings and diagnostic yield was carried out. RESULTS: MRI was abnormal in 109 cases. It never resulted in any patient care modification. 55 malformations, 12 cases of cerebral atrophy, 7 cases of white matter disease and 2 patients with phakomatose were identified. Myelination delay (26 cases), increased signal of posterior white matter on T2-weighted images (9 cases) or widened Virchow-Robin spaces (3 cases) were frequently encountered, but it remained unclear whether they represented normal variants or true abnormalities. Post ischemic lesions were identified in 10 cases. Frequency of abnormal studies was significantly lower in children with developmental delay and behavioral disorders than in patients with other clinical presentation. CONCLUSION: Diagnostic yield of cerebral MRI can justify its performance by comparison to other imaging modalities. It should be correlated with other investigations performed in a specialized unit. Its main interest is for classification and research. Risk of sedation or anesthesia should also be taken in account. Risk can be lowered using adequately equipped MR units and organizing procedures in collaboration with anesthesiologists.

Adolescent↗

Histopathologic variability of the congenital nephrotic syndrome.

Five cases of congenital nephrotic syndrome (CNS) were studied with light microscopic (LM), transmission electron microscopic (TEM) and immunofluorescence microscopic (IFM) techniques. All patients, aged between three weeks and one year, had a nephrotic syndrome (NS) and died soon after the diagnosis. Three patients showed mesangial proliferation, thinning of glomerular basement membranes (GBM) and no deposits by IFM or TEM. They probably represent variants of the Finnish type of CNS. One patient had diffuse sclerosis of the mesangium, irregular and thickened GBMs and electron dense deposits, consistent with the diagnosis of diffuse mesangial sclerosis (DMS). The fifth case also showed changes suggestive of DMS, but was clinically atypical, with an early onset and rapid course. Our findings confirm the morphologic heterogeneity of the CNS and point out the difficulties of classification based on clinical and histological manifestations.

Female↗

Recent progress in the histological typing of human breast tumours.

Recently, new concepts in the histological typing of human mammary carcinomas have been proposed. The basic subdivision into the two major categories, i.e. ductal and lobular carcinomas, remains valid. In so far as lobular carcinomas are concerned, histological variants have been recognized: invasive carcinoma without an in situ component as well as solid and signet-ring cell forms. Special types of duct carcinoma have been defined with more accuracy, those deserving special attention being, scirrhous, tubular and secretory forms. The validity of the diagnosis of so-called apocrine carcinoma is discussed, as is the place of mucus-secreting tumours in the histologic classification.

Adenocarcinoma↗

[Ubiquitous and rare blood group related antigens and their significance in serology].

The present classification of some factors which are counted among the ubiquitary private and familiar as well as the rare and rarest antigens is shown. The classification of these antigens is often difficult. The distinguishing and systematisation of rare and rarest antigens requires of the investigator much consistency, responsibility and consideration as well as universal calmness so as not to bring too much confusion into an already broad palete of private and familiar antigens. Some antigens are frequently rearranged into already existing systems or even become independent. The authors boldly point out the variants of the ABO and Rh systems in which we meet with some ever so rare antigens, but which, as we know, are nevertheless not counted among the rare, in the sense of present comprehension. The practical value is pointed out and the need of improving our knowledge of ubiquitary, rare, familiar and private antigens in order to be able to master certain complications which these antigens would provoke in serology. In difficulties it is necessary to consult an experienced specialist in a well organised laboratory which possesses several different and reliable serums, the erythrocyte tests and a lot of experience.

Blood Group Antigens↗

Differentiation of Potato virus Y strains using improved sets of diagnostic PCR-primers.

Potato virus Y (PVY) is one of the most important viruses of potato world-wide, several strain groups are recognized. In the past two decades, novel PVY variants have appeared causing necrotic symptoms on potato tubers. Implicated are two groups of recombinant strains: PVY(N)W and PVY(NTN), and NA-PVY(NTN). While the first two are recombinants between PVY-N- and O-strains the latter is a recombinant between an N-strain and an unknown PVY strain or other Potyvirus. Available biological and molecular data on PVY suggest that classification of PVY strains has to be revised. Some drawbacks have been found with recently published primers used in RT-PCR based differentiation of PVY strains as some defined isolates could not be identified correctly. Consequently we developed new primers using both recently available sequences and newly generated complete sequences of PVY strains. The reliability of these newly developed primers and procedures was successfully demonstrated on nearly 100 biologically and serologically characterised PVY isolates.

Amino Acid Sequence↗

A genome-wide analysis of the shared genetic risk architecture of complex neurological and psychiatric disorders.

Although neurological and psychiatric disorders have historically been considered to reflect distinct pathogenic entities, recent findings suggest shared pathophysiological mechanisms. However, the extent to which these heritable disorders share genetic influences remains unclear. Here we performed a comprehensive analysis of genome-wide association study data, involving nearly 1 million cases across ten neurological diseases and ten psychiatric disorders, to compare their common genetic signal and biological associations. Using complementary statistical tools, we demonstrate that a large set of common genetic variants impacts the risk of multiple neurological and psychiatric disorders, even in the absence of genetic correlations. Furthermore, genome-wide association studies on psychiatric disorders consistently implicate neuronal biology, whereas neurological diseases are associated with diverse neurobiological processes. Together, this study elucidates the genetic relationship between complex neurological and psychiatric disorders, indicating a larger degree of genetic pleiotropy than previously recognized. The findings have implications for disease classification, precision medicine and clinical practice.

Humans↗

Study on the early-onset variant of benign childhood epilepsy with occipital paroxysms otherwise described as early-onset benign occipital seizure susceptibility syndrome.

PURPOSE: We studied the early-onset variant of benign childhood epilepsy with occipital paroxysms (EVBCEOP) proposed by Panayiotopoulos, to confirm whether his five criteria are sufficient to delineate EVBCEOP as a new epileptic syndrome, as well as to predict a good outcome prospectively at the time of the first examination. SUBJECTS: The subjects were 649 children with localization-related epilepsies (LREs) observed in our hospital for >4 years. METHODS: We applied the International Classification of Epilepsies and Epileptic Syndromes to the 649 patients and identified patients who had EVBCEOP from among those with nonspecific idiopathic LRE. The inclusion criteria were to satisfy all five criteria and all but one criterion (i.e., either ictal vomiting or occipital EEG paroxysms). We were blind as to the outcomes and selected patients who satisfied the following three of the five criteria at the time of the first examination, (a) normal development before the onset, (b) epilepsy onset age between 2 and 8 years, and (c) occipital EEG foci. We attempted to determine whether the outcome can be predicted prospectively, and whether the presence or absence of ictal vomiting affects the prognosis. RESULTS: We identified 19 patients who satisfied all five criteria, 22 who exhibited all but occipital EEG foci, and 21 who exhibited all but ictal vomiting. The incidence of status convulsivus was higher in those with ictal vomiting than in those without ictal vomiting (p < 0.05). Interictal EEG performed every 6 months showed shifting and multiplication of EEG foci in 42 and 52% of all subjects, respectively. We identified 57 patients, 42 (74%) of whom were in remission by age 12 years. The number of patients who experienced remission did not differ significantly between those with (76%, n = 25) and without (72%, n = 32) ictal vomiting (p > 0.05). CONCLUSIONS: Nosologically, EVBCEOP appears to constitute the earliest form of idiopathic epileptic syndrome different from classic BCEOP. However, its clinical spectrum, ranging from the absence of ictal vomiting to a combination of extraoccipital and multifocal EEG foci, is broad, such that further prospective study is expected to reveal the exact prerequisite criteria for determining the border of this epileptic syndrome and for clarifying the clinical spectrum within this syndrome.

Adolescent↗