ABSTRACT
The abundance of Lp(a) protein holds significant implications for the risk of cardiovascular disease (CVD), which is directly impacted by the copy number (CN) of KIV-2, a 5.5 kbp sub-region. KIV-2 is highly polymorphic in the population and accurate analysis is challenging. In this study, we present the DRAGEN KIV-2 CN caller, which utilizes short reads. Data across 166 WGS show that the caller has high accuracy, compared to optical mapping and can further phase ~50% of the samples. We compared KIV-2 CN numbers to 24 previously postulated KIV-2 relevant SNVs, revealing that many are ineffective predictors of KIV-2 copy number. Population studies, including USA-based cohorts, showed distinct KIV-2 CN, distributions for European-, African-, and Hispanic-American populations and further underscored the limitations of SNV predictors. We demonstrate that the CN estimates correlate significantly with the available Lp(a) protein levels and that phasing is highly important.
ABSTRACT
Peripheral-type benzodiazepine binding sites (PBRs) are ubiquitous in mammalian tissues. However, the physiological role of PBRs has not yet been clarified. In this study we characterized a saturable and high affinity binding site for [3H]Pk 11195 (isoquinoline carboxamide derivative) on human lymphocytes and different lymphoma cell lines. Binding parameters of the human T-lymphoma cell line CCRF-CEM came closest to values for lymphocyte binding. Thus, the CCRF-CEM cell line appears to be a suitable lymphocyte cell model for further study of PBRs. To evaluate the pharmacological specificity of binding to human lymphocytes and CCRF-CEM cells we investigated the potency of different ligands to displace [3H]Pk 11195 from its binding site. Pk 11195 was found to be the most potent inhibitor followed by 4'-chlorodiazepan (Ro5-4864) and diazepam (range of inhibition constants from 6.7 x 10(-9) M to 3.6 x 10(-7) M), whereas ligands specific for the central-type receptor like clonazepam and flumazenil had no displacing potency in the tested concentration range (10(-10)-10(-4) M). Since it was assumed that PBRs might be involved in the regulation of cell growth and differentiation, we studied the influence of PBR ligands on cell growth and survival using a quantitative colorimetric assay (MTT). Ligands which bind selectively to PBRs inhibited cell multiplication in vitro. However, half-effective concentrations (EC50) were in the micromolar range and above therapeutic in vivo concentrations (range of EC50 values from 2.4 x 10(-5) M to 1.5 x 10(-4) M). Clonazepam and flumazenil had no inhibiting potency in the tested concentration range (10(-10)-10(-4) M). Although the difference between values for displacing potency and ability to inhibit cell multiplication cannot be explained as yet, it is interesting that all PBR-ligands followed the same sequence in displacing [3H]Pk 11195 and inhibiting cell multiplication and that central type ligands were ineffective in both assays. This association suggest a mediating role of PBR binding in cell growth.
Subject(s)
Lymphocytes/metabolism , Lymphoma/metabolism , Receptors, GABA-A/metabolism , Adult , Binding Sites , Cell Division , Cell Line/drug effects , Dose-Response Relationship, Drug , Female , Humans , Isoquinolines/pharmacokinetics , Male , Receptors, GABA-A/drug effects , TritiumABSTRACT
Modulators for the reversal of multidrug resistance such as R-verapamil and B8509-035, a dihydropyridine, effectively overcome multidrug resistance in vitro and are currently undergoing clinical trial. One problem with their use is the application protocol; the question as to whether they should be given by continuous administration or in sequential doses in combination with the cytotoxic drugs has to be addressed. Therefore, we examined the influence of the exposure time and the sequence of modulator administration on the active transport of the fluorescent dye rhodamine 123 (R123), a substrate for the P-glycoprotein, in the resistant lymphoblastoid cell line VCR1000 and the parental nonresistant cell line CCRF-CEM. Our results demonstrate the importance of coadministration of R-verapamil and the cytotoxic agent for the modulation of multidrug resistance, whereas the exposure sequence does not seem to be such an essential parameter in the case of B8509-035. This observation should be considered for the further design of clinical studies.
Subject(s)
Antineoplastic Agents/pharmacology , Dihydropyridines/pharmacology , Precursor Cell Lymphoblastic Leukemia-Lymphoma/pathology , Rhodamines/metabolism , Verapamil/pharmacology , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Antineoplastic Agents/administration & dosage , Cell Differentiation/drug effects , Dihydropyridines/administration & dosage , Dose-Response Relationship, Drug , Drug Interactions , Drug Resistance , Flow Cytometry , Humans , Membrane Glycoproteins/biosynthesis , Tumor Cells, Cultured , Verapamil/administration & dosageABSTRACT
The use of implanted central venous catheters with ports has become almost irreplaceable in clinical treatment of cancer patients with advanced disease. It improves the patient's quality of life and simplifies medical treatment. For safe use during outpatient follow-up treatment, possible complications such as wound infections, thrombosis or even pulmonary embolism have to be considered. The incidence of catheter-associated thrombosis in cancer patients has been estimated as 25-66% in screening evaluations. Clinical symptoms were present in only in 6-28% of all patients with diagnosed thromboses. We report on a patient with metastatic malignant melanoma, who developed a deep arm vein thrombosis two weeks after port implantation. The prognosis of deep arm vein thrombosis is determined by the development of pulmonary embolism (8-36%). In such cases, early diagnosis and treatment are essential for the patient's survival.
Subject(s)
Catheterization, Central Venous/adverse effects , Pulmonary Embolism/etiology , Pulmonary Embolism/prevention & control , Subclavian Vein/diagnostic imaging , Venous Thrombosis/etiology , Venous Thrombosis/prevention & control , Humans , Male , Melanoma/complications , Melanoma/secondary , Melanoma/therapy , Middle Aged , Radiography , Skin Neoplasms/complications , Skin Neoplasms/therapyABSTRACT
With the help of the tetracycline resistance transposon Tn10 in and around the mgl genes the gene cir was mapped. cir is 80% cotransducible with mgl by P1 transduction. The sequence of the surrounding markers in clockwise order was established as: cdd fpk cir mgl gyrA. The direction of transcription in cir was determined as clockwise on the Escherichia coli chromosome. The gene product of cir, an outer membrane receptor for colicin I, is not part of the mgl operon. It is not regulated by D-fucose, the inducer of the mgl system and mutants defective in cir are unimpaired in the uptake of substrates of the mgl-dependent transport system.
Subject(s)
Chromosomes, Bacterial/ultrastructure , Colicins/pharmacology , Drug Resistance, Microbial , Escherichia coli Proteins , Escherichia coli/genetics , Genes, Bacterial , Receptors, Cell Surface , Receptors, Immunologic/genetics , Chromosome Mapping , DNA Transposable Elements , Transcription, GeneticABSTRACT
A 41-year old male patient presented with yellowish papules on the elbows, upper arms, back and the buttocks, as well as yellow streaks in the palmar creases. Laboratory examination revealed Fredrickson type IIb hyperlipidemia. Histologic changes were consistent with eruptive xanthomas. Treatment was started with HMG-CoA reductase inhibitors and dietary measures were taken to lower serum levels of cholesterol and triglycerides. Hyperlipidemia is the cause of eruptive xanthomas and strongly increases the risk of cardiovascular diseases.
Subject(s)
Xanthomatosis/diagnosis , Adult , Biopsy , Combined Modality Therapy , Diet, Fat-Restricted , Humans , Hydroxymethylglutaryl-CoA Reductase Inhibitors/therapeutic use , Hyperlipoproteinemia Type II/diagnosis , Hyperlipoproteinemia Type II/pathology , Hyperlipoproteinemia Type II/therapy , Male , Skin/pathology , Xanthomatosis/pathology , Xanthomatosis/therapyABSTRACT
A 54-year-old female working as a vegetable farmer presented with painful pruritic skin lesions on both hands. Physical examination showed a hyperkeratotic fissured eczema. RAST, prick- and patch testing revealed type I and IV hypersensitivity to spinach, ruccola, and chives, so that a protein contact dermatitis was diagnosed.
Subject(s)
Chive/adverse effects , Dermatitis, Allergic Contact/etiology , Dermatitis, Occupational/etiology , Mustard Plant/adverse effects , Spinacia oleracea/adverse effects , Dermatitis, Allergic Contact/diagnosis , Dermatitis, Allergic Contact/drug therapy , Dermatitis, Occupational/diagnosis , Dermatitis, Occupational/drug therapy , Female , Glucocorticoids/therapeutic use , Humans , Middle Aged , Skin TestsABSTRACT
The silver staining technique for demonstration of argyrophilic nucleolar organizer regions (AgNORs) was examined for their usefulness in routine clinical diagnosis for discrimination of benign breast diseases from malignant ones and for prognostic purposes. We examined the number of AgNORs in 56 cases of malignant breast lesion and 20 cases of benign breast disease, using routinely fixed material. We found a clear-cut difference between benign breast diseases as compared with breast carcinomas where the mean AgNOR count was significantly higher, whereas no statistically significant correlation was observed between AgNOR counts and survival rate of human breast cancer patients.
Subject(s)
Breast Neoplasms/pathology , Carcinoma/pathology , Fibroadenoma/pathology , Fibrocystic Breast Disease/pathology , Nucleolus Organizer Region/pathology , Adenocarcinoma, Mucinous/pathology , Breast Neoplasms/mortality , Carcinoma/mortality , Carcinoma, Ductal, Breast/pathology , Carcinoma, Lobular/pathology , Carcinoma, Medullary/pathology , Epithelium/pathology , Female , Fibrocystic Breast Disease/mortality , Humans , Nucleolus Organizer Region/ultrastructure , Receptors, Estrogen/analysis , Receptors, ProgesteroneABSTRACT
We report here on a new sensitive and highly specific DNA staining technique which we have called sulpho-DNA staining. DNA staining is based on a sulphonylation reaction of 2'-deoxycytidine or cytidine that takes place in the 6th position of cytosine with ensuing immunodetection of the sulphonylated DNA. The specificity of DNA staining is introduced by the use of an antibody recognizing only modified DNA but not modified RNA, by recourse to an additional acid hydrolysis step which destroys RNA but not DNA. We describe here the optimal conditions for the sulphonylation of DNA using O-methylhydroxylamine and metabisulphite as reactants. The new DNA stain labels all nuclei in either normal human tissue or in tumor cells. For nuclear DNA the staining signal is higher for the sulpho-DNA staining than for the Feulgen staining for nuclear DNA. This new DNA staining technique is suitable for use on tissue sections as well as on cytosmears.