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Biomedical subjects

J De Jong

Publications and source records attributed to J De Jong.

At least 19 recordsLinked to original sources

OCT4: biological functions and clinical applications as a marker of germ cell neoplasia.

Germ cell tumours (GCTs) are a heterogeneous group of neoplasms, which develop in the gonads as well as in extragonadal sites, that share morphological patterns and an overall good prognosis, owing to their responsiveness to current surgical, chemotherapeutic, and radiotherapeutic measures. GCTs demonstrate extremely interesting biological features because of their close relationships with normal embryonal development as demonstrated by the pluripotentiality of some undifferentiated GCT variants. The similarities between GCTs and normal germ cell development have made it possible to identify possible pathogenetic pathways in neoplastic transformation and progression of GCTs. Genotypic and immunophenotypic profiles of these tumours are also useful in establishing and narrowing the differential diagnosis in cases of suspected GCTs. Recently, OCT4 (also known as OCT3 or POU5F1), a transcription factor that has been recognized as fundamental in the maintenance of pluripotency in embryonic stem cells and primordial germ cells, has been proposed as a useful marker for GCTs that exhibit features of pluripotentiality, specifically seminoma/dysgerminoma/germinoma and embryonal carcinoma. The development of commercially available OCT4-specific antibodies suitable for immunohistochemistry on paraffin-embedded specimens has generated increasing numbers of reports of OCT4 expression in a wide variety of gonadal and extragonadal GCTs. OCT4 immunostaining has been shown to be a sensitive and specific marker for seminomatous/(dys)germinomatous tumours and in embryonal carcinoma variants of non-seminomatous GCTs, whether in primary gonadal or extragonadal sites or in metastatic lesions. Therefore, OCT4 immunohistochemistry is an additional helpful marker both in the differential diagnosis of specific histological subtypes of GCTs and in establishing a germ cell origin for some metastatic tumours of uncertain primary. OCT4 expression has also been reported in pre-invasive conditions such as intratubular germ cell neoplasia, unclassified (IGCNU) and the germ cell component of gonadoblastoma. Additionally, OCT4 immunostaining shows promise as a useful tool in managing patients known to be at high risk for the development of invasive GCTs.

Biomarkers, Tumor↗

Attempt to detect evidence for tick-borne encephalitis virus in ticks and mammalian wildlife in The Netherlands.

To investigate if tick-borne encephalitis virus (TBEV) is present in mammalian wildlife species or ticks in The Netherlands, serum samples and ticks were tested for TBEV antibodies and TBEV RNA, respectively. Serum samples were collected from wild boar (666), deer (13), fox (399), and rodents (90), and were tested for TBEV antibodies, using ELISA, and SN test or HI test. Over a period of 4 years, a total of 906 ticks was collected from seven regions in The Netherlands. In four different regions, this was done on a monthly basis and during four consecutive summers. All ticks were tested for TBEV RNA by RT-PCR. TBEV antibody was detected by ELISA in two (0.5%) sera of foxes and 49 (7%) sera of wild boar, but not confirmed by HI or SNT. TBEV RNA was not detected in any of 906 ticks. It was concluded that there is no real evidence for a TBEV reservoir in ticks or wildlife in The Netherlands.

Animals↗

Cloning of the chrysanthemum UEP1 promoter and comparative expression in florets and leaves of Dendranthema grandiflora.

To attain high transgene expression in petal tissue of ray florets of chrysanthemum an endogenous ubiquitin extension protein (UEP1) promoter was cloned and tested with the beta-glucuronidase (GUS) reporter gene. Expression levels were compared with four heterologous promoters: chalcone synthase (chs-A) and zinc finger transcription factor (EPF2-5) from petunia, eceriferum (CER6) from Arabidopsis and multicystatin (PMC) from potato. The comparison of the expression levels of the different constructs in ray florets, disc florets, and leaves is presented. The highest mean expression in petal tissue of ray and disc florets was conferred by the UEP1 promoter, followed by CER6 and EPF2-5. The UEP1 promoter in ray florets confers over 50-fold enhancement in expression as compared to CaMV 35S-based promoters.

Amino Acid Sequence↗

Novel mutations in the 7-dehydrocholesterol reductase gene of 13 patients with Smith--Lemli--Opitz syndrome.

Smith--Lemli--Opitz syndrome (SLOS) is caused by mutations in the DHCR7 gene leading to deficient activity of 7-dehydrocholesterol reductase (DHCR7; EC 1.3.1.21), the final enzyme of the cholesterol biosynthetic pathway, resulting in low cholesterol and high concentrations of its direct precursor 7-dehydrocholesterol in plasma and tissues. We here report mutations identified in the DHCR7 gene of 13 children diagnosed with SLOS by clinical and biochemical criteria. We found a high frequency of the previously described IVS8--1 G > C splice acceptor site mutation (two homozygotes, eight compound heterozygotes). In addition, 13 missense mutations and one splice acceptor mutation were detected in eleven patients with a mild to moderate SLOS-phenotype. The mutations include three novel missense mutations (W182L, C183Y, F255L) and one novel splice acceptor site mutation (IVS8--1 G > T). Two patients, homozygous for the IVS8--1 G > C mutation, presented with a severe clinical phenotype and died shortly after birth. Seven patients with a mild to moderate SLOS-phenotype disclosed compound heterozygosity of the IVS8--1 G > C mutation in combination with different novel and known missense mutations.

Child↗

Role of common gene variations in the molecular pathogenesis of short-chain acyl-CoA dehydrogenase deficiency.

ABSTRACT Short-chain acyl-CoA dehydrogenase (SCAD) deficiency is considered a rare inherited mitochondrial fatty acid oxidation disorder. Less than 10 patients have been reported, diagnosed on the basis of ethylmalonic aciduria and low SCAD activity in cultured fibroblast. However, mild ethylmalonic aciduria, a biochemical marker of functional SCAD deficiency in vivo, is a common finding in patients suspected of having metabolic disorders. Based on previous observations, we have proposed that ethylmalonic aciduria in a small proportion of cases is caused by pathogenic SCAD gene mutations, and SCAD deficiency can be demonstrated in fibroblasts. Another - much more frequent - group of patients with mild ethylmalonic aciduria has functional SCAD deficiency due to the presence of susceptibility SCAD gene variations, i.e. 625G>A and 511C>T, in whom a variable or moderately reduced SCAD activity in fibroblasts may still be clinically relevant. To substantiate this notion we performed sequence analysis of the SCAD gene in 10 patients with ethylmalonic aciduria and diagnosed with SCAD deficiency in fibroblasts. Surprisingly, only one of the 10 patients carried pathogenic mutations in both alleles, while five were double heterozygotes for a pathogenic mutation in one allele and the 625G>A susceptibility variation in the other. The remaining four patients carried only either the 511C>T or the 625G>A variations in each allele. Our findings document that patients carrying these SCAD gene variations may develop clinically relevant SCAD deficiency, and that patients with even mild ethylmalonic aciduria should be tested for these variations.

Acyl-CoA Dehydrogenase↗

Assessment of antibodies to double-stranded DNA induced in rheumatoid arthritis patients following treatment with infliximab, a monoclonal antibody to tumor necrosis factor alpha: findings in open-label and randomized placebo-controlled trials.

OBJECTIVE: To compare the incidence of anti-double-stranded DNA (anti-dsDNA) antibodies in rheumatoid arthritis (RA) patients receiving either single or multiple doses of a chimeric anti-tumor necrosis factor alpha (anti-TNFalpha) antibody or placebo infusions, with or without methotrexate, in open-label, randomized, placebo-controlled trials. METHODS: Multiple sera obtained from 156 patients before and after treatment with infliximab and from 37 patients treated with placebo infusions were tested for anti-dsDNA antibodies by 3 methods: Crithidia luciliae indirect immunofluorescence test (CLIFT), a commercial Farr assay (Ortho Diagnostics radioimmunoassay [RIA]) in which the antigen source is mammalian DNA, and a Farr assay employing 125I-labeled circular plasmid DNA (Central Laboratory of The Netherlands Red Cross Blood Transfusion Service [CLB] RIA). Patients with positive findings on the CLIFT were also tested for antibodies to histones (H1-H5) and chromatin and for IgM rheumatoid factors (IgM-RFs). RESULTS: None of the RA patients had a serum sample that was positive for anti-dsDNA antibodies by the CLIFT prior to infliximab therapy. Of the 22 patients who developed a positive CLIFT result, 11 (7% of 156 exposed to infliximab) also had positive findings on the Ortho RIA at a concentration of >10 units/ml and another 8 (5%) were positive at a concentration of >25 units/ml. In all but 1 patient, the anti-dsDNA antibodies were solely of the IgM isotype. Only 1 patient had detectable anti-dsDNA antibodies by the CLB RIA. All sera containing anti-dsDNA by the CLIFT contained antibodies to chromatin, and sera from 2 patients also contained antibodies to histones. IgM-RF titers showed a significant reduction following infliximab therapy in these 22 patients. One patient developed anti-dsDNA antibodies of IgG, IgA, and IgM isotype and had positive results on both Farr assays (peaking at 22 weeks and resolving by 54 weeks); this was associated with a reversible lupus syndrome. CONCLUSION: Anti-dsDNA antibodies of IgM class are induced by infliximab therapy; the frequency is dependent on the assay method used. Only 1 of the 156 patients who were treated with infliximab developed a self-limiting clinical lupus syndrome; that patient developed high titers of anti-dsDNA antibodies of IgG, IgM, and IgA class, as detected by the CLIFT and by 2 different Farr assays.

Adult↗

Effects on chick performance and nutrient digestibility of an endo-xylanase added to a wheat- and rye-based diet in relation to fat source.

1. A study with growing chicks investigated the effects of an inclusion of an endo-xylanase preparation (LYXASAN) to a wheat- and rye-based diet on performance and nutrient digestibility in relation to the fat source. 2. The basal diet contained 500 g wheat and 100 g rye/kg of diet. The basal diet was supplemented with either 65 g soya oil/kg or 60 g blended animal fat and 5 g soya oil/kg. 3. Endo-xylanase added to the soya oil diet did not affect weight gain, but there was a numerical improvement in food conversion efficiency which was not statistically significant. When the endo-xylanase preparation was added to the blended animal fat diet, both weight gain and food utilisation were improved by 9.5% and 6.0%, respectively (P < 0.05). 4. Digestibilities of organic matter, crude fat, crude fibre and NFE were not significantly affected by adding endo-xylanase to the soya oil diet. However, when endo-xylanase was included in the blended animal fat diet, digestibility of organic matter, crude fat, crude fibre and NFE were improved (P < 0.05). The improvement in fat digestibility was the most pronounced, amounting to 9.4%. Nitrogen retention and metabolisable energy content were improved significantly by the addition of an endo-xylanase to the animal fat diet (P < 0.05), by 6.6% and 6.5% respectively. 5. From the results of this study, it can be concluded that the effects on chick performance and nutrient digestibility of a dietary endo-xylanase in a wheat- and rye-based diet are influenced to a considerable degree by the type of fat in the diet.

Animal Feed↗

Replacement value of betaine for DL-methionine in male broiler chicks.

The effect of DL-methionine and betaine supplementation on growth performance of 2,400 male broilers in the age period of 1 to 38 d, and on carcass composition of a subsample of 384 birds was examined. Three dose levels of DL-methionine (0, 0.05, and 0.10%) and two doses of betaine (0 and 0.04%) were supplemented in different combinations to methioninedeficient diets. Two types of diets were fed as starters and growers: either corn-soybean diets or practical diets typical for the Dutch broiler industry. All diets were fortified with 220 ppm choline in order to avoid a deficiency in methyl groups. Increasing DL-methionine supplementation significantly improved daily weight gain and feed conversion efficiency. Supplemental betaine did not affect bird growth. Betaine slightly improved feed conversion in diets without supplemental DL-methionine, but did not affect this parameter in diets with added DL-methionine. Breast meat yield was significantly increased by about 1.5 percentage points by the addition of 0.05% DL-methionine, whereas 0.04% betaine only tended to increase breast meat yield in the range of 0.3 to 0.6 percentage points. The type of diet did not have any effect on the responses obtained. In summary, there was no evidence for betaine to spare DL-methionine as an essential amino acid supplement in broiler diets.

Analysis of Variance↗

V3 loop sequence analysis of seven HIV type 1 group O isolates phenotyped in peripheral blood mononuclear cells and MT-2 cells.

HIV-1-infected individuals from which syncytium-inducing (SI) viruses are isolated most often progress more rapidly to AIDS than individuals carrying only non-syncytium-inducing (NSI) viruses. The syncytium-inducing capacity of virus isolates is commonly determined in conjunction to replication in MT-2 cells. Comparison of HIV-1 env sequences and a site-directed mutagenesis study have indicated that the presence of a positively charged amino acid at position 11 or 25 in the V3 loop is minimally required for the SI capacity of HIV-1 subtype B viruses. Studies have also shown a similar correlation between positively charged signature amino acids in the V3 loop and syncytium formation in MT-2 cells for HIV-1 subtypes A, D, and E. In the present study virus phenotype was determined and compared to the V3 loop sequence of seven HIV-1 group O isolates. Three of the HIV-1 group O isolates showed the NSI/non-MT-2 tropic phenotype and two showed the SI/MT-2 tropic phenotype, whereas two isolates presented an uncommon NSI/MT-2 tropic phenotype. The V3 loop of the two SI/MT-2 tropic isolates had a high net positive charge and contained a positively charged amino acid at position 11 or 25. The V3 loop of the two NSI/MT-2 tropic isolates had a low net positive charge and contained a single positively charged amino acid at position 37.

Amino Acid Sequence↗

A fairly conserved epitope on the hemagglutinin of influenza A (H3N2) virus with variable accessibility to neutralizing antibody.

A monoclonal antibody LMBH5 was derived from mice which had been immunized with A/Victoria/3/75 (H3N2)-type recombinant, secreted hemagglutinin (HA), and were subsequently challenged with a potentially lethal dose of X31 [A/Aichi/2/68 (H3N2) x A/PR/8/34 (H1N1)] virus. LMBH5 reacted strongly with the native and low-pH-induced conformations of the HA of A/Aichi (X31 strain) and A/Victoria (X47 strain), but very weakly with the native structure of the HA of A/Philippines/2/82 (X79 strain) and not at all with the HA of A/Guizhou/54/89 H3 (NIB25 strain). However, the acid-induced conformations of the latter two viruses were recognized by LMBH5. The antibody prevented infection of MDCK cells with X31 and X47, whereas X79 virus was partially neutralized by LMBH5. X31 monoclonal escape variants had single amino acid substitutions (Ser 227-->Pro) near the interface. The data obtained suggest that the neutralizing LMBH5 reacts with a fairly conserved epitope of influenza A (H3N2) virus, which as a result of antigenic drift becomes inaccessible in the native state of the HA.

Animals↗

Phase I clinical and pharmacokinetic study of oral etoposide phosphate.

PURPOSE: To determine the bioavailability (F) and the pharmacokinetic profile of both etoposide and its prodrug, etoposide phosphate, after oral and intravenous administration of etoposide phosphate, and to determine the maximum-tolerable dose (MTD) of oral etoposide phosphate administered daily for 5 consecutive days every 3 weeks. In addition, we sought to develop and validate two limited-sampling models (LSMs) to predict the etoposide area under the curve (AUC) 24 hours after administration of oral and intravenous etoposide phosphate. PATIENTS AND METHODS: In the F part of the study, patients were assessed for pharmacokinetic studies after one oral and one intravenous administration of the same dose of etoposide phosphate. Etoposide phosphate and etoposide plasma concentrations were assayed by high-performance liquid chromatography (HPLC). To develop LSMs after oral and intravenous administration, patients were randomized between the training and validation data sets. In the phase I part of the study, which followed the F part, the dose of etoposide phosphate was escalated from 50 mg/m2/d for etoposide equivalents for 5 days to 220 mg/m2/d for 5 days. RESULTS: Forty adult patients with solid tumors or lymphoma entered the study and 35 were assessable for toxicity. The MTDs were defined as 175 mg/m2 and 220 mg/m2 in previously treated and untreated patients, respectively. Neutropenia was dose-limiting, with high interpatient variability. Within 15 minutes after intravenous administration, etoposide phosphate was no longer detectable in plasma, and it was never detectable after oral administration. Plasma concentrations and pharmacokinetic parameters of etoposide following etoposide phosphate were comparable to those reported for etoposide. The relative F (mean +/- SD) of etoposide after oral etoposide phosphate was 76 +/- 27%, with a range of 37% to 144%. CONCLUSION: The clinical and pharmacokinetic results of this study confirm the prodrug hypothesis of etoposide phosphate. Although firm conclusions cannot be drawn, the F of oral etoposide phosphate seems to be comparable to or only slightly better than that of oral etoposide.

Administration, Oral↗

Requirement of the laying hen for sulfur amino acids.

Two experiments with individually caged laying hens were conducted to determine the requirement for TSAA. In Experiment 1, the corn-soybean basal diet contained .48% TSAA to which increasing dose levels of DL-methionine were added providing at the highest supplemental level .645% TSAA. The experimental diets were fed for 12 wk, covering the early stage of laying from 25 to 37 wk of age. In Experiment 2, a corn-soybean basal diet containing of .51% TSAA was used along with DL-methionine supplements to determine TSAA requirement during the entire laying cycle of 52 wk (25 to 77 wk of age). The highest supplemental level of DL-methionine in this experiment provided .76% TSAA. The TSAA requirement was found to be higher for maximum efficiency of feed utilization than for obtaining maximum egg production. Based on feed conversion efficiency and at an egg mass yield of 55 g per hen-day, the requirement for TSAA was estimated to be about 740 mg per hen-day, of which about 440 mg was methionine, throughout a laying period of 52 wk. It was calculated that the estimated TSAA requirement was equivalent to approximately 660 mg true digestible SAA per hen-day.

Amino Acids, Sulfur↗

Disturbances of hypothalamic-pituitary-adrenal axis functioning during ethanol withdrawal in six men.

OBJECTIVE: Excessive exposure to glucocorticoids can have neurotoxic effects. The behavioral, cognitive, and neurochemical changes observed following the cessation of heavy drinking, therefore, may be associated with disturbances of the hypothalamic-pituitary-adrenal (HPA) axis. To investigate HPA axis disturbances during the ethanol withdrawal syndrome, the authors examined diurnal changes in plasma cortisol in six alcohol-dependent men following the abrupt discontinuation of alcohol intake. METHOD: Plasma cortisol concentrations were quantified every 30 minutes for 24 hours in the early stage (1 day after cessation) and the middle to late stage (3 days after cessation) of the ethanol withdrawal syndrome as well as after the resolution of acute symptoms (8 days or more after cessation). RESULTS: Plasma cortisol concentrations were almost twice as high during acute withdrawal as they were following recovery. The duration of the cortisol diurnal cycle on the first day of withdrawal was negatively correlated with the severity of withdrawal. CONCLUSIONS: There is a marked activation of the HPA axis associated with the ethanol withdrawal syndrome. The authors hypothesize that this activation may account for some of the signs and symptoms of acute and subacute withdrawal. They discuss the potential long-term physiological effects of the episodic increases in cortisol associated with repeated episodes of ethanol withdrawal. The alterations in cortisol rhythmicity during early withdrawal may also have clinical implications.

Adult↗

The detection of anti-Ro/SS-A and anti-La/SS-B antibodies. A comparison of counterimmunoelectrophoresis with immunoblot, ELISA, and RNA-precipitation assays.

The presence in serum of anti-Ro/SS-A and/or anti-La/SS-B autoantibodies is a characteristic of autoimmune diseases such as Sjögren's syndrome and systemic lupus erythematosus. To evaluate different assays currently available for the detection of these antibodies 50 sera were tested using the four different assay methods: counterimmunoelectrophoresis (CIE), RNA precipitation assay, immunoblotting technique and ELISA. The RNA-precipitation assay showed the highest sensitivity and specificity. The CIE for the detection of anti-Ro/SS-A antibodies gave comparable results whereas the Ro/SS-A ELISA and Ro/SS-A or HeLa immunoblot showed lower sensitivities (96% and 80% respectively). Sensitivity was even lower (66%) when only reactivity towards the 60 kDa Ro/SS-A protein was considered. The ELISA for the detection of anti-La/SS-B antibodies showed a sensitivity of 98%, the immunoblotting technique of 86% and the CIE only 67%. The high sensitivity of the La/SS-B ELISA went together with a low specificity of 14%. We conclude from these data that for the detection of anti-Ro/SS-A and anti-La/SS-B antibodies the RNA precipitation assay shows the highest sensitivity and highest specificity. For routine screening purposes the CIE is the most convenient and reliable assay to detect anti-Ro/SS-A antibodies. For the detection of anti-La/SS-B antibodies the immunoblot corresponds most closely to the RNA precipitation assay.

Antibodies, Antinuclear↗

A cerebrospinal fluid study of the pathophysiology of panic disorder associated with alcoholism.

In order to investigate the neurochemistry of panic disorder in alcoholics, we measured various cerebrospinal fluid (CSF) parameters in subjects with both conditions and compared them with an age- and sex-matched population of alcoholics and normal controls. When height, age and weight were covaried, subjects with panic disorder had higher levels of B-endorphin in CSF. There were no differences in other CSF measures between the groups. Alcoholics with panic disorder had higher plasma MHPG concentrations compared with alcoholics without panic disorder but these were not statistically different from controls.

Adult↗

Cerebrospinal fluid monoamine metabolites in alcoholic patients who attempt suicide.

Reduced cerebrospinal fluid (CSF) levels of the serotonin metabolite 5-hydroxyindoleacetic acid have been reported to be commonly associated with suicidal behaviour. Alcoholics are known to often manifest suicidal behaviour. Therefore, we compared cerebrospinal fluid levels of monoamine metabolites in alcoholics who had (n = 20) and had not (n = 108) attempted suicide and healthy volunteers (n = 30). There were no significant differences among the 3 groups for CSF levels of either 5-hydroxyindoleacetic acid, the dopamine metabolite homovanillic acid, norepinephrine, or the norepinephrine metabolite 3-methoxy-4-hydroxyphenylglycol.

Adult↗