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1.
Pol J Vet Sci ; 24(2): 271-279, 2021 Jun.
Article in English | MEDLINE | ID: mdl-34250787

ABSTRACT

The etiology of Postpartum dysgalactia syndrome (PDS) includes stress οn preparturition and constipation associated with low water intake or low fiber intake. The aim of this study was to investigate the effects of a raw crude fibre concentrate (Arbocel®) on sow's metabolism and performance. 100 sows from a farm suffering from PDS, were divided into two groups, with equal distribu- tion of their parity (1 to 5 parity): a) T1 group (control group): 50 sows were fed with regular gestation feed (GF), pre-farrowing feed (PFF), and lactation feed (LF), b) T2 group: 50 sows were fed with regular GF, PFF and LF supplemented with topdress Arbocel® from 104th day of gestation until 7th day of lactation). Health parameters [faeces score (FS), PDS score (PDSS), body condition score (BCS)], performance parameters and liter characteristics were recorded. Blood samples were collected from 25 sows / group (5 sows per parity) 24 h after birth of last piglet and on 14th day of lactation for the evaluation of insulin, leptin and ghrelin levels in the serum, using commercial ELISA kits. In T2 group, BCS at farrowing (p⟨0.001), FS (p=0.001) and PDSS (p=0.003) were improved significantly. The number of piglets stillborn and dead due to crushing decreased (p=0.001), while the number of liveborn (p=0.016) and weaned piglets (p=0.001) increased in T2 group. Moreover, in T2 group, the BW of piglets at weaning was higher (p⟨0.001). A significant increase of insulin (p=0.032) and leptin (p=0.032) levels in serum was noticed in T2 group 24 h after farrowing. In conclusion, the supplementation of extra crude fibre in breeding stock with PDS problems due to nutritional imbalance has beneficial effects on their health and performance.


Subject(s)
Diet/veterinary , Dietary Fiber/pharmacology , Energy Metabolism/drug effects , Lactation Disorders/veterinary , Swine Diseases/prevention & control , Animal Feed/analysis , Animal Nutritional Physiological Phenomena , Animals , Dietary Fiber/administration & dosage , Lactation Disorders/prevention & control , Postpartum Period , Swine
2.
J Transl Med ; 19(1): 151, 2021 04 15.
Article in English | MEDLINE | ID: mdl-33858454

ABSTRACT

BACKGROUND: Pharmacogenomics describes the link between gene variations (polymorphisms) and drug responses. In view of the implementation of precision medicine in personalized healthcare, pharmacogenetic tests have recently been introduced in the clinical practice. However, the translational aspects of such tests have been limited due to the lack of robust population-based evidence. MATERIALS: In this paper we present a novel pharmacogenetic panel (iDNA Genomics-PGx-CNS or PGx-CNS), consisting of 24 single nucleotide polymorphisms (SNPs) on 13 genes involved in the signaling or/and the metabolism of 28 approved drugs currently administered to treat diseases of the Central Nervous System (CNS). We have tested the PGx-CNS panel on 501 patient-derived DNA samples from a southeastern European population and applied biostatistical analyses on the pharmacogenetic associations involving drug selection, dosing and the risk of adverse drug events (ADEs). RESULTS: Results reveal the occurrences of each SNP in the sample and a strong correlation with the European population. Nonlinear principal component analysis strongly indicates co-occurrences of certain variants. The metabolization efficiency (poor, intermediate, extensive, ultra-rapid) and the frequency of clinical useful pharmacogenetic, associations in the population (drug relevance), are also described, along with four exemplar clinical cases illustrating the strong potential of the PGx-CNS panel, as a companion diagnostic assay. It is noted that pharmacogenetic associations involving copy number variations (CNVs) or the HLA gene were not included in this analysis. CONCLUSIONS: Overall, results illustrate that the PGx-CNS panel is a valuable tool supporting therapeutic medical decisions, urging its broad clinical implementation.


Subject(s)
Pharmaceutical Preparations , Pharmacogenetics , Central Nervous System , DNA Copy Number Variations/genetics , Humans , Precision Medicine
3.
J Matern Fetal Neonatal Med ; 26(16): 1576-80, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23544842

ABSTRACT

OBJECTIVE: The aim of this study was to examine whether resistin is present in second trimester amniotic fluid from trisomy 21 (also known as Down's syndrome) pregnancies and whether its concentration differs compared with euploid pregnancies. METHODS: The study cohort consisted of 58 women in the mid-trimester of pregnancy who underwent amniocentesis for prenatal diagnosis, 31 of whom carried a single fetus with diagnosed trisomy 21 (study group) and the rest with normal karyotype (control group, n = 27). Groups were matched for maternal and gestational age. Levels of resistin in amniotic fluid were measured by a commercially available enzyme-linked immunosorbent assay (ELISA) kit. RESULTS: Resistin was detected in all amniotic fluid samples. Its median concentration in the second trimester amniotic fluid of trisomy 21 pregnancies (2.1 ng/ml) was statistically significantly lower (p value <0.001) in comparison with that in euploid pregnancies (3.3 ng/ml). CONCLUSIONS: Resistin is a physiologic constituent of second trimester amniotic fluid. Lower levels of amniotic fluid resistin in pregnancies with trisomy 21 may reflect altered metabolic pathways in utero that could possibly be related with phenotypic features of the syndrome.


Subject(s)
Amniotic Fluid/metabolism , Down Syndrome/metabolism , Pregnancy Trimester, Second/metabolism , Resistin/metabolism , Adult , Amniotic Fluid/chemistry , Case-Control Studies , Cohort Studies , Down Syndrome/diagnosis , Female , Gestational Age , Humans , Predictive Value of Tests , Pregnancy , Resistin/analysis
4.
Mediators Inflamm ; 2012: 138971, 2012.
Article in English | MEDLINE | ID: mdl-22489108

ABSTRACT

OBJECTIVE: We investigated whether the concentration of the glycoprotein fetuin A is altered in the second trimester amniotic fluid of trisomy 21 pregnancies compared with euploid pregnancies. METHODS: 25 pregnancies with an extra chromosome 21 were matched for maternal and gestational age with 25 pregnancies with normal karyotype. Levels of fetuin A in amniotic fluid were measured by a commercially available enzyme-linked immunosorbent assay (ELISA) kit. RESULTS: The median concentration of fetuin A in amniotic fluid of trisomy 21 pregnancies (5.3 ng/ml) was statistically significantly lower (P value = 0.008) compared with that in euploid pregnancies (6.8 ng/mL). CONCLUSION: Lower levels of fetuin A in trisomy 21 may indicate an association with altered metabolic pathways in this early stage that could potentially be associated with features of the syndrome, such as growth restriction or impaired osteogenesis.


Subject(s)
Amniotic Fluid/metabolism , Down Syndrome/genetics , Fetus/metabolism , Pregnancy Trimester, Second/metabolism , alpha-2-HS-Glycoprotein/metabolism , Adult , Chromosomes, Human, Pair 21/genetics , Enzyme-Linked Immunosorbent Assay , Female , Humans , Pregnancy
6.
Ocul Immunol Inflamm ; 9(2): 103-9, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11449326

ABSTRACT

PURPOSE: To describe a case of herpes simplex virus type 2 (HSV-2) acute retinal necrosis syndrome (ARN) in a 13-year-old immunocompetent girl. METHODS: Polymerase chain reaction (PCR), cultures, flow cytometry, and cytology were performed on the vitreous sample. RESULTS: Both PCR studies and vitreous cultures revealed HSV-2 as the cause of ARN. Flow cytometry showed CD4+, CD8+, and natural killer cells. The visual outcome of the patient was 20/200. CONCLUSION: Successful culture of HSV-2 from the vitreous specimen in a patient with ARN proved HSV-2 to be one of the causes of ARN. The successful culture of HSV-2 has not been previously reported.


Subject(s)
Eye Infections, Viral , Herpes Genitalis , Herpesvirus 2, Human/isolation & purification , Retinal Necrosis Syndrome, Acute/virology , Adolescent , Antibodies, Viral/analysis , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , DNA, Viral/analysis , Eye Infections, Viral/diagnosis , Eye Infections, Viral/virology , Female , Flow Cytometry , Herpes Genitalis/diagnosis , Herpes Genitalis/virology , Herpesvirus 2, Human/genetics , Herpesvirus 2, Human/immunology , Humans , Immunoglobulin M/analysis , Killer Cells, Natural/immunology , Polymerase Chain Reaction , Retinal Detachment/diagnostic imaging , Retinal Detachment/virology , Retinal Necrosis Syndrome, Acute/diagnosis , Ultrasonography , Visual Acuity , Vitreous Body/virology
7.
Dermatology ; 196(1): 171-5, 1998.
Article in English | MEDLINE | ID: mdl-9557256

ABSTRACT

Cytochromes P-450 are a supergene family of enzymes involved in the metabolism of a wide range of endogenous and foreign compounds. The existing genetic variations of the distinct isozymes lead to interindividually different metabolic capacity. Since vitamin A, endogenous retinoids and their natural metabolites are morphogenic for the sebaceous gland, we investigated the polymorphisms of cytochrome P-450 1A1, as being one of the most active isozymes involved in their interconversion. From the known mutations, two were investigated; an additional cleavage site for MspI in the 3'-flanking region identified as a thymine-to-cytosine transition 1,194 bp downstream of exon 7 (m1) and an adenine-to-guanine transition at position 4889 in exon 7 (m2). We studied 96 acne patients for m1 and m2 mutations by restriction fragment length polymorphism and allele-specific polymerase chain reaction, respectively, and compared the results with 408 reference individuals. No statistically significant difference was found in the distribution of m2 alleles; the frequency was 3.13 and 3.06% of the alleles, respectively (odds ratio = 1.02, confidence limits 0.41-2.52, p = 0.96). In contrast, a trend to an overrepresentation of m1 alleles in acne patients was observed; allele frequency was 8.33 in the patients and 6.99% in the control subjects, respectively (odds ratio 1.21, 95% confidence limits 0.68-2.16, p = 0.52). As the m1 mutation might define a marker for alterations on regulatory sites, the biological efficacy of natural retinoids could be greatly impaired by their rapid metabolism to inactive compounds. The resulting deficit of active natural retinoids may lead to abnormal sebocyte differentiation and hyperkeratinization of the follicular canal implicating the development of acne in some patients.


Subject(s)
Acne Vulgaris/genetics , Cytochrome P-450 CYP1A1/genetics , Polymorphism, Genetic , Alleles , Humans , Mutation , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length
8.
Am J Ophthalmol ; 123(5): 699-702, 1997 May.
Article in English | MEDLINE | ID: mdl-9152082

ABSTRACT

PURPOSE: To describe a case of retinal perivasculitis in an immunocompetent patient with systemic herpes simplex infection. METHODS: Polymerase chain reaction amplifications were performed for aqueous and blood samples using primers specific for the following members of the herpesvirus family: cytomegalovirus, Epstein-Barr virus, herpes simplex virus (types 1 and 2), and varicella-zoster virus. The patient was placed on intravenous acyclovir and systemic corticosteroids. RESULTS: A positive polymerase chain reaction signal was found only for herpes simplex virus type 1. Vision in the left eye improved from light perception to 20/25, and signs of retinal perivasculitis resolved. CONCLUSION: The use of molecular diagnostic modalities in clinical practice may aid in determining infectious etiologies in patients with atypical clinical manifestations.


Subject(s)
Eye Infections, Viral/diagnosis , Herpes Simplex/diagnosis , Immunocompetence , Retinal Vein/pathology , Retinitis/diagnosis , Vasculitis/diagnosis , Acyclovir/therapeutic use , Antiviral Agents/therapeutic use , Aqueous Humor/virology , DNA, Viral/analysis , Eye Infections, Viral/drug therapy , Eye Infections, Viral/etiology , Female , Fluorescein Angiography , Fundus Oculi , Herpes Simplex/drug therapy , Herpes Simplex/etiology , Herpesvirus 1, Human/genetics , Herpesvirus 1, Human/isolation & purification , Humans , Middle Aged , Polymerase Chain Reaction , Retinitis/drug therapy , Retinitis/virology , Vasculitis/drug therapy , Vasculitis/virology , Viremia/virology
9.
Am J Hum Genet ; 57(3): 581-92, 1995 Sep.
Article in English | MEDLINE | ID: mdl-7668286

ABSTRACT

The polymorphic arylamine N-acetyltransferase (NAT2; EC 2.3.1.5) is supposed to be a susceptibility factor for several drug side effects and certain malignancies. A group of 844 unrelated German subjects was genotyped for their acetylation type, and 563 of them were also phenotyped. Seven mutations of the NAT2 gene were evaluated by allele-specific PCR (mutation 341C to T) and PCR-RFLP for mutations at nt positions 191, 282, 481, 590, 803, and 857. From the mutation pattern eight different alleles, including the wild type coding for rapid acetylation and seven alleles coding for slow phenotype, were determined. Four hundred ninety-seven subjects had a genotype of slow acetylation (58.9%; 95% confidence limits 55.5%-62.2%). Phenotypic acetylation capacity was expressed as the ratio of 5-acetylamino-6-formylamino-3-methyluracil and 1-methylxanthine in urine after caffeine intake. Some 6.7% of the cases deviated in genotype and phenotype, but sequencing DNA of these probands revealed no new mutations. Furthermore, linkage pattern of the mutations was always confirmed, as tested in 533 subjects. In vivo acetylation capacity of homozygous wild-type subjects (NAT2*4/*4) was significantly higher than in heterozygous genotypes (P = .001). All mutant alleles showed low in vivo acetylation capacities, including the previously not-yet-defined alleles *5A, *5C, and *13. Moreover, distinct slow genotypes differed significantly among each other, as reflected in lower acetylation capacity of *6A, *7B, and *13 alleles than the group of *5 alleles. The study demonstrated differential phenotypic activity of various NAT2 genes and gives a solid basis for clinical and molecular-epidemiological investigations.


Subject(s)
Arylamine N-Acetyltransferase/genetics , Genetic Linkage , Alleles , Base Sequence , Genotype , Humans , Molecular Sequence Data , Mutation , Phenotype , Polymerase Chain Reaction , White People/genetics
10.
Clin Pharmacol Ther ; 56(6 Pt 1): 626-34, 1994 Dec.
Article in English | MEDLINE | ID: mdl-7995004

ABSTRACT

Polymorphic liver arylamine N-acetyltransferase (NAT2; EC 2.3.1.5) has been suggested as a susceptibility factor for both insulin-dependent diabetes mellitus (IDDM) and non-insulin-dependent diabetes mellitus. Previous studies reported an overrepresentation of phenotypically fast acetylators among patients with diabetes. With use of an allele-specific nested polymerase reaction, the NAT2 genotypes were determined in 165 clinically well-controlled patients with IDDM and 107 reference children aged from 3 to 18 years. Wild-type and mutated alleles (mutation 1 diagnosed by presence of cytosine at position 341 instead of thymine; M2 by adenine at 590 instead of guanine, M3 by adenine at 857 instead of guanine) were distributed equally in both groups. Genotypes coding fast acetylation (homozygous wild-type and heterozygous wild-type with one of the mutations) were detected in 40.6% and 36.6% of children with IDDM and reference children, respectively (odds ratio, 1.19; 95% confidence limits, 0.70 to 2.04). In 66 children with IDDM and 54 reference children the NAT2 genotype was checked by conventional sulfamethazine (sulfadimidine) phenotyping. There were only five discrepant cases, indicating that NAT2 genotyping enables correct prediction of NAT2 phenotype in about 95% of tested individuals. The fast acetylator genotype could not be established as a host factor for IDDM susceptibility in children.


Subject(s)
Arylamine N-Acetyltransferase/genetics , Diabetes Mellitus, Type 1/enzymology , Acetylation , Adolescent , Alleles , Base Sequence , Child , Child, Preschool , Diabetes Mellitus, Type 1/genetics , Female , Genotype , Heterozygote , Humans , Male , Molecular Sequence Data , Mutation , Odds Ratio , Phenotype , Polymorphism, Genetic , Predictive Value of Tests
11.
Cancer Res ; 54(15): 4103-11, 1994 Aug 01.
Article in English | MEDLINE | ID: mdl-8033143

ABSTRACT

Glutathione S-transferase M1 (GSTM1) is a foreign compound-metabolizing enzyme with a heritable complete lack of activity in about 50% of Caucasians. GSTM1 deficiency may predispose individuals to urinary bladder cancer. Thus, a hospital-based case-control study was performed with 296 patients with bladder cancer and 400 controls, investigating this GSTM1 deficiency in relation to environmental risk factors and types of bladder cancer. Frequencies of the GSTM1 gene deletion (genotype, GSTM1*0/0) and of the allele variants A (mu) and B (psi) of the GSTM1-active trait were determined using an internal standard-controlled polymerase chain reaction technique. Moreover, in all patients GSTM1 expression was quantified in blood by an immunoassay. Of the cases, 59.1% had the GSTM1*0/0 genotype, in contrast to 50.7% of the controls (odds ratio, 1.40; 95% confidence limits, 1.02-1.92; P = 0.017). The odds ratio after adjustment for age and gender by logistic regression analysis was 1.54 (95% confidence limits, 1.12-2.13). Occupational risk was defined as previous employment in occupations with known increased bladder cancer risk, but the impact of GSTM1*0/0 was not significantly different in individuals with risk jobs versus those without. The greater proportion of the GSTM1-deficient individuals in the group with cancer was due to a lower frequency of carriers of GSTM1A. The odds ratio for the subgroup of individuals with the GSTM1B phenotype versus carriers of the GSTM1A phenotype in cases versus controls was 1.65 (95% confidence limits, 0.976-2.78; two-tailed Fisher's exact P = 0.057). Analysis of functional GSTM1 activity in a subset of 370 blood samples with the model substrate trans-stilbene oxide confirmed the genetic results and showed that 9 of 10 individuals with mu/psi heterodimers (genotype, GSTM1*A/B) had activities above the median of all genetically GSTM1-active individuals (24 pmol/min/1 x 10(6) lymphocytes; P < 0.01), indicating a gene dose relationship for GSTM1. GSTM1 expression in the urinary bladder endothelium detected by immunoassay and immunohistology corresponded to the genotype of the patients. It may be concluded from this study that the heritable GSTM1 deficiency is responsible for 17% (etiological fraction; 95% confidence limits, 2-30%) of bladder cancer cases.


Subject(s)
Coenzymes/deficiency , Glutathione Transferase/deficiency , Occupational Exposure/adverse effects , Smoking/adverse effects , Urinary Bladder Neoplasms/enzymology , Urinary Bladder/enzymology , Aged , Alleles , Base Sequence , Case-Control Studies , Coenzymes/chemistry , Coenzymes/genetics , Female , Genotype , Glutathione Transferase/chemistry , Glutathione Transferase/genetics , Humans , Male , Middle Aged , Molecular Sequence Data , Occupations , Odds Ratio , Phenotype , Polymerase Chain Reaction , Urinary Bladder Neoplasms/pathology
12.
Clin Investig ; 72(3): 240-8, 1994 Feb.
Article in English | MEDLINE | ID: mdl-7912124

ABSTRACT

Genetic differences in the metabolism of carcinogens may codetermine individual predisposition to cancer. Cytochrome P-4501A1 (CYP1A1) metabolically activates precarcinogens in cigarette smoke, such as benzo(a)pyrene, which is also an inducer of CYP1A1. Two point mutations have been reported, m1 in the 3'-flanking region (6235T to C), and m2 within exon 7 (4889A to G), the latter leading to an isoleucine to valine exchange. In the Japanese population m1 and m2 are correlated with lung cancer, suggesting an increased susceptibility to cigarette smoking related lung cancer. We studied 142 lung cancer and 171 reference patients in an ethnically homogeneous German group for m1 and m2 mutations by restriction fragment length polymorphism and allele-specific polymerase chain reaction, respectively. No statistically significant difference was found in the distribution of m1 alleles between lung cancer and controls; the frequency was 8.5% and 7.3% of the alleles, respectively (odds ratio = 1.17). A trend to an overrepresentation of m1 alleles was observed among 52 squamous cell carcinoma patients (odds ratio = 1.65). In contrast, the frequency of m2 alleles in lung cancer patients was twofold higher (6.7%) than in the reference group (3.2%; odds ratio = 2.16; 95% confidence limits 0.96-5.11, P = 0.033); the odds ratio of m2 alleles in squamous cell carcinoma was 2.51 (95% confidence limits 0.85-7.05, P = 0.05). There was a close genetic linkage of m2 to m1 (10 of 11 reference patients), but a significantly higher number of cancer patients showed no linkage compared to the controls (odds ratio = 8.89, 95% confidence limits 0.83-433, P = 0.04).(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Carcinoma, Bronchogenic/genetics , Cytochrome P-450 Enzyme System/genetics , Exons , Genes , Lung Neoplasms/genetics , Neoplasm Proteins/genetics , Point Mutation , Polymorphism, Restriction Fragment Length , Adult , Aged , Aged, 80 and over , Alleles , Base Sequence , Biotransformation , Carcinogens/pharmacokinetics , Carcinoma, Bronchogenic/enzymology , Female , Gene Frequency , Genetic Predisposition to Disease , Humans , Lung Diseases/enzymology , Lung Diseases/genetics , Lung Neoplasms/enzymology , Male , Middle Aged , Molecular Sequence Data , Odds Ratio , White People/genetics
13.
Cancer Res ; 53(5): 1004-11, 1993 Mar 01.
Article in English | MEDLINE | ID: mdl-8439946

ABSTRACT

Glutathione S-transferase class mu (GSTM1) is known to detoxify certain carcinogens or their activated metabolites. In a previous study using phenotyping methods, individuals genetically devoid of this enzyme activity were significantly overrepresented among lung cancer patients compared to controls, suggesting that this trait is a risk factor for lung cancer. Here, GST class mu status has been determined both pheno- and genotypically, i.e., (a) by ex vivo measurement of trans-stilbene oxide conjugation in lymphocytes, (b) by GSTM1 quantification in blood using an immunoassay, and (c) by the application of polymerase chain reaction to genomic DNA with characterization of an inactivating mutation responsible for the null allele and a G/C single base allelic variance corresponding to the polymorphism of GSTM1 isoenzymes mu and psi, respectively. One hundred seventeen lung cancer patients and 155 control patients were studied. The two groups were of German origin and were similar with respect to age, sex, smoking history, and catchment area. In about 97% of cases, the three methods of assigning activity type of GSTM1 gave corresponding results. By genotype, 55 of 117 lung cancer patients (47.0%) and 73 of 155 control patients (47.1%) were GSTM1 active. The control group was confirmed by analysis of GSTM1 genotype in 200 further, independently studied reference patients; 101 of them were GSTM1 active (50.5%). Thus, the hypothesis of heritable GSTM1 deficiency as a host factor predisposing to lung cancer proved inappropriate. Detailed analysis of subgroups with respect to smoking habits, age, and sex failed to reveal an impact of GST class mu genotype on lung cancer risk. Among the total of 272 patients studied, 36 individuals carried at least one psi allele; however, no unexpected frequency distribution was observed.


Subject(s)
Glutathione Transferase/genetics , Isoenzymes/genetics , Lung Neoplasms/enzymology , Adult , Aged , Aged, 80 and over , Alleles , Base Sequence , Female , Genotype , Glutathione Transferase/analysis , Humans , Isoenzymes/analysis , Lung Neoplasms/etiology , Lung Neoplasms/pathology , Male , Middle Aged , Molecular Sequence Data , Phenotype , Risk Factors
14.
Clin Investig ; 70(3-4): 307-19, 1992.
Article in English | MEDLINE | ID: mdl-1355678

ABSTRACT

Epidemiological studies suggested a protective effect of certain phenotypes of polymorphic foreign-compound-metabolizing enzymes in some types of cancer. Poor metabolizers (PM) of debrisoquine 4-hydroxylase (cytochrome P-450IID6, CYP2D6) were found to be underrepresented among patients with lung cancer. Recent advances in molecular genetic characterization of CYP2D6, glutathione S-transferase (GST) class Mu, and arylamine N-acetyltransferase enabled genotypical determination of mutant alleles in lung cancer patients. Restriction fragment length polymorphism (RFLP) with a cDNA gene probe of CYP2D6 was analyzed in 79 lung cancer patients who were phenotyped with debrisoquine. Mutant alleles were detected by allele-specific polymerase chain reaction (PCR). In the same individuals, genotype of GST class Mu was analyzed by PCR and correlated with ex vivo activity of glutathione conjugation towards trans-stilbene oxide. RFLP patterns allowed discrimination between the slow and fast genotype of N-acetyltransferase as well as the heterozygotes. Three phenotypical PMs of debrisoquine (3.8%) were confirmed by PCR and RFLP. No PM could be unambiguously recognized only by RFLP patterns. The PMs were characterized by PCR and RFLP as carriers of the 29B/29B (n = 1), 29A/29B (n = 1), and 29A/44 (n = 1) mutant alleles. Higher debrisoquine hydroxylase activities were found in the homozygous EMs, who possess two active genes, as compared to heterozygous EMs, who have only one active gene. The patients with phenotypically impaired GST Mu activity were confirmed as such by PCR. A complete correspondence between phenotyping of N-acetyltransferase (with caffeine) and genotyping was found. The new genetic techniques proved to be powerful tools for molecular-epidemiological studies aimed at establishing host factors of cancer susceptibility.


Subject(s)
Arylamine N-Acetyltransferase/genetics , Cytochrome P-450 Enzyme System/genetics , Glutathione Transferase/genetics , Lung Neoplasms/genetics , Mixed Function Oxygenases/genetics , Neoplasm Proteins/genetics , Acetylation , Aged , Alleles , Cytochrome P-450 CYP2D6 , DNA Mutational Analysis , Female , Genes , Genetic Predisposition to Disease , Genotype , Humans , Lung Neoplasms/enzymology , Male , Middle Aged , Phenotype , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Risk Factors , Smoking
15.
Biochem Pharmacol ; 43(3): 647-50, 1992 Feb 04.
Article in English | MEDLINE | ID: mdl-1540219

ABSTRACT

Glutathione S-transferase (GST) class Mu activity was determined in 145 unrelated hospital patients in Berlin by measuring their conjugation activity towards the specific substrate trans-stilbene oxide (TSO) with two substrate concentrations (50 and 250 microM) in homogenates prepared from lymphocytes. Eighty individuals (55.2%) had an activity lower than 10 pmol/min/10(6) lymphocytes and were classified as GST class Mu deficient. In 142 of 145 cases, phenotype was confirmed by the results of a genotyping procedure using the polymerase chain reaction technique. Two fragments of 273 and about 650 bp including one and two introns, respectively, could always be amplified from genomic DNA in individuals with high GST class Mu activity and could not be amplified in persons with impaired glutathione-TSO conjugation activity. This indicates that persons with low activity carry a large deletion mutation within the GST class Mu gene. The enzymatically determined antimode between low and high activity determined as 10 pmol/min/1 million lymphocytes in the assay with 50 microM TSO could be clearly confirmed by genotyping.


Subject(s)
Chromosome Deletion , Glutathione Transferase/deficiency , Glutathione/metabolism , Isoenzymes/deficiency , Lung Neoplasms/enzymology , Stilbenes/metabolism , Adult , Aged , Aged, 80 and over , Base Sequence , Berlin , Female , Germany/ethnology , Glutathione Transferase/blood , Glutathione Transferase/genetics , Humans , Isoenzymes/blood , Isoenzymes/genetics , Lymphocytes/enzymology , Male , Middle Aged , Molecular Sequence Data , Mutation , Phenotype , Polymerase Chain Reaction
16.
Klin Wochenschr ; 66 Suppl 11: 87-97, 1988.
Article in English | MEDLINE | ID: mdl-2846954

ABSTRACT

Bronchogenic carcinoma is closely associated with cigarette smoking although additional environmental or individual factors might regulate a person's susceptibility to that disease. To further define such risk factors, the prevalence of the genetic debrisoquine 4-hydroxylation deficiency was determined before therapeutic intervention in 270 lung cancer patients. Nineteen homozygous carriers of this defect (poor metabolizers) were found (7.0%, 95% confidence limits 4.3%-10.8%), a number being lower than 30 out of 270 reference patients (11.1%, 95% confidence limits 7.6%-15.5%). The odds ratio of 0.61 was of marginal statistical significance (P = 0.067). Subdividing the collective according to histology revealed a trend towards underrepresentation of poor metabolizers especially among patients with adenocarcinoma (1 out of 37, P = 0.086) and among young patients not older than 50 years (none out of 32, P = 0.028). All poor metabolizers (PMs) in the cancer group were smokers. In 18 patients the phenotype assignment was confirmed by a second test several weeks after surgical or other treatment. In 220 of the lung cancer patients the N-acetyltransferase polymorphism was evaluated by means of the molar ratio of 5-acetylamino-6-formylamino-3-methyluracil and 1-methylxanthine in urine after ingestion of caffeine (coffee). There were 111 (50.5%) slow acetylators and 109 (49.5%) fast acetylators. A statistically significant clustering of either phenotype after stratification according to histology, or debrisoquine hydroxilator status was lacking. Moreover, there was no difference in the ratio of both phenotypes as compared to the reference collective of 245 patients (53.5% slow and 46.5% fast acetylators). As a third genetic host factor the AB0 blood group frequencies were evaluated in 263 lung cancer patients. The frequency ratio of A/O was significantly higher as compared to 41,423 blood donors (odds ratio 1.37, 95% confidence limits 1.02-1.84, P less than 0.05). A/O tended to be especially high in young patients not older than 50 years. The ratio B/O in bronchial cancer was significantly higher than expected. The results suggest that the debrisoquine hydroxilator status might have an impact on an individual's susceptibility to lung cancer. This association is either a weak one and/or is restricted to certain histological cancer types or to patients with certain characteristics. The acetylator phenotype could not be established as a risk factor, whereas AB0 blood groups seem to influence lung cancer susceptibility.


Subject(s)
ABO Blood-Group System/genetics , Acetyltransferases/genetics , Arylamine N-Acetyltransferase/genetics , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Small Cell/genetics , Cytochrome P-450 Enzyme System/genetics , Lung Neoplasms/genetics , Mixed Function Oxygenases/genetics , Phenotype , Acetylation , Caffeine , Cytochrome P-450 CYP2D6 , Debrisoquin/pharmacokinetics , Humans , Polymorphism, Genetic , Risk Factors
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