Biomedical subjects

D Janssen

Publications and source records attributed to D Janssen.

At least 37 records · Page 2Linked to original sources

PubMed2003

Rapid detection of cucumber vein yellowing virus by tissue-print hybridisation with digoxigenin-labelled cDNA probes.

Hybridisation of tissue prints with nonradioactive cDNA probes was developed to detect cucumber vein yellowing virus (CVYV) in cucurbit plants. Results showed irregular distribution of the virus within cucumber, zucchini or melon plants without defined tropism for a specific tissue. Therefore, reliable diagnosis of CVYV requires analysis of tissue prints from at least five different plant sites. This detection procedure allows rapid analysis of large numbers of plants and it can be useful for epidemiological studies of CVYV and to control virus spread via eradication of early foci.

Cucumis sativus↗

PubMed2003

Comparison between human pharmacokinetics and imaging properties of two conjugation methods for 99mTc-annexin A5.

Annexin A5 (AnxA5) is a protein with high affinity for phosphatidyl serine, a phospholipid exposed on the cell surface during apoptosis. This phenomenon has been used for determination of cell death after myocardial infarction. To evaluate the potential of (99m)Tc-AnxA5 for in vivo scintigraphy of apoptotic cells, the pharmacokinetics and imaging properties of two radiopharmaceuticals, (99m)Tc-(n-1-imino-4-mercaptobutyl)-AnxA5 (I-AnxA5) and (99m)Tc-(4,5-bis(thioacetamido)pentanoyl)-AnxA5 (B-AnxA5), were studied. I-AnxA5 was administered intravenously to seven patients and one healthy volunteer, and B-AnxA5 was administered to 12 patients. All patients in the pharmacokinetic study had myocardial disease. Additionally, imaging was performed in a patient with acute myocardial infarction, as well as in three patients with different malignancies. The plasma concentration, excretion and biodistribution of (99m)Tc-AnxA5 were measured, as well as levels of AnxA5 antigen. The kinetic data of both radiopharmaceuticals in plasma fitted a two-compartment model. Both preparations had similar half-lives, but a different distribution over the two compartments. Plasma levels of AnxA5 antigen showed a broad variation. Both radiopharmaceuticals accumulated in the kidney, liver and gut. B-AnxA5 was excreted significantly faster than I-AnxA5. Both compounds can be used for imaging of the head/neck region, the thorax and the extremities. B-AnxA5 has a faster clearance and a lower radiation dose. Imaging of apoptosis in the abdomen will be difficult with both radiopharmaceuticals, and especially with B-AnxA5 because of its faster appearance in the gut.

Adult↗

PubMed2001

The effect of lameness on milk production in dairy cows.

Data were collected prospectively for 1.5 yr on two New York dairy farms to investigate the effect of lameness on milk production. The numbers of study cows (percentages treated at least once for lameness) in each herd were 1796 (52%) and 724 (40%), respectively. Lame cows were identified and treated by farm employees or a professional hoof trimmer. Weekly averages of total milk production per day were recorded based on automated milk weight measurements at each milking. The effect of lameness on milk production was analyzed separately for each herd using repeated measures ANOVA. In both herds, milk production decreased significantly for cows diagnosed lame. Milk production was 1.5 kg/d lower > or = 2 wk after lameness compared with cows that had not yet been diagnosed lame in the current lactation in the larger herd. In the second herd, milk production of lame cows was 0.8 kg/d lower in the first and second wk after lameness and 0.5 kg/d lower > or = 3 wk after diagnosis. The decrease in milk production associated with lameness was larger for cows in second or greater lactation and for more severe cases. In one herd, the decrease in milk production was greater for cows with sole ulcers or foot abscesses than for foot rot or foot warts. Cows with abscesses or foot rot tended to have larger decreases in milk production in the other herd. The inconsistent results between farms may have resulted from differences in the relative frequencies of specific causes of lameness in the two herds and in the way lame cows were identified and defined for the study.

Analysis of Variance↗

PubMed2001

Biodistribution and dosimetry of 99mTc-BTAP-annexin-V in humans.

The purpose of this study was to determine the biodistribution and the associated radiation dose of technetium-99m 4,5-bis(thioacetamido)pentanoyl-annexin-V (99mTc-Apomate), a tracer proposed for the study of apoptosis. Eight patients (including two females) with normal kidney and liver functions were included in the study. An activity of 580 +/- 90 MBq of 99mTc-Apomate was injected intravenously, immediately followed by a dynamic study of 30 frames of 1 min each. At about 1 h, 4 h and 20 h p.i., whole-body scans were acquired. All activity distributions were measured using a dual-head gamma camera. Before injection of activity, a transmission scan with a cobalt-57 flood source had been performed to determine patient attenuation. Blood samples were taken every 10 min during the first hour after injection, and at about 4 and 20 h. Urine and faeces were collected during the first 20 h. Organ uptake was estimated after correction for body background activity, attenuation and scatter. Residence times were calculated from the dynamic and whole-body studies and used as input in the Mirdose 3.1 program to obtain organ doses and effective dose. It was found that radioactivity strongly accumulated in the kidneys and the liver [at 70 min p.i., 28% +/- 8% and 20% +/- 4% of the injected dose (ID), respectively]. Uptake in the target tissues (lymphomas or heart) was negligible from a dosimetric point of view. Extrapolating data from the first 20 h, one finds that approximately 73% of the ID will be excreted in the urine, and 27% in the faeces. The biological half-life of the activity in the total body was 16 +/- 7 h. Some organ doses +/- standard deviation (SD) in microGy/MBq were: kidneys 63 +/- 22, urinary bladder 20 +/- 6, spleen 15 +/- 3, liver 13 +/- 3, upper large intestine 12 +/- 6, lower large intestine 8 +/- 4, testes 6 +/- 2 and red bone marrow 4 +/- 0.7. The effective dose was 7.6 +/- 0.5 microSv/MBq, corresponding to a total effective dose of 4.6 +/- 0.3 mSv for a nominal injected activity of 600 MBq. In conclusion, 99mTc-Apomate has a high uptake in the kidneys and liver--in fact a factor of 1.3-1.6 higher than that found for the previously studied 99mTc-(n-1-imino-4-mercaptobutyl)-annexin-V. The biological half-life is shorter, however, but still long compared with the physical half-life of 99mTc. The faster appearance of activity in the intestines may preclude imaging of apoptosis in the abdomen. The effective dose is within the lower range of values reported for typical 99mTc compounds.

Adult↗

PubMed1997

Immunomodulation by hydatid cyst fluid toxin (Echinococcus granulosus).

Since the experimental infection by hydatid cysts (Echinococcus granulosus) in mice causes immunomodulation of the host, the effects of hydatid fluid (HF) and fractions of HF were compared in vitro and in vivo. Fractions of HF were obtained using ammonium sulphate precipitation, chloroform/methanol extraction and thin-layer chromatography (TLC). HF proved to be toxic to murine peritoneal macrophages in vitro, and when macrophages were incubated with the different fractions of HF, most toxicity was found in a single TLC-purified fraction with an adjuvant-like effect on the production of specific antibodies against bovine albumin and human red blood cells in mice. Treatment of mice with the toxin caused a drop in the percentage of peripheral blood lymphocytes. Flow-cytometric analysis showed that T-cells from toxin-treated mice had lower membrane-CD3, CD4 and CD8 density, and higher percentages of CD8+ splenocytes and CD4+ thymocytes expressing CD25. The toxin caused a down-regulation of CD4 and CD8 expression on thymocytes in vitro, that was dependent on the presence of macrophages. The results may attribute to these toxins a role in the host-parasite relationship of hydatidosis.

Animals↗

PubMed1997

Specific recognition of hydatid cyst antigens by serum IgG, IgE, and IgA using western blot.

Diagnosis of hydatid disease in humans relies on the detection of specific antibodies against antigens of the metacestode from Echinococcus granulosus. The specificity and sensitivity of current immunological techniques based on specific serum IgG rely on the way antigens are purified. We used Western immunoblotting to detect specific IgG, IgE, and IgA antibodies in serum from patients with hydatid disease using either crude antigen preparations (total hydatid fluid), purified fractions enriched in Antigens 5 and B, and glycoproteins from hydatid fluid. Depending on whether crude HF or purified antigen fractions were used, IgG and IgE recognized specifically low-to-medium MW bands between 12 and 42 kDa. IgA recognized specifically 110 kDa band in crude hydatid fluid and in the glycoprotein fraction of hydatid fluid, and a 42 kDa band in all antigen samples used. Besides the advantage of detecting specific IgA in crude hydatid fluid, these results offer the possibility of simplifying future immunological tests if specific secretory IgA can be similarly detected.

Adult↗

PubMed1996

Serologic recognition of hydatid cyst antigens using different purification methods.

The specificity and sensitivity of enzyme-linked immunosorbent assays (ELISA) and Western immunoblot assays in detecting antibodies in serum from patients suffering cystic hydatid disease (Echinococcus granulosus) are compared using either crude antigen preparations (total sheep hydatid fluid and homogenates of protoscoleces), purified fractions enriched in Antigens 5 and B, and glycoproteins from hydatid fluid. Polyprotein bands of 12-14, 20, and 34 kDa, when purified from hydatid fluid by applying changes in the ionic strength, yielded a sensitive (95%) immunodiagnostic test that was also extremely specific (100%) when assayed with sera from noninfected humans and from patients suffering from other parasitic diseases. However, subjecting hydatid fluid to chromatography through a concanavilin A column rendered a 42 kDa band that was sensitive (95%) as well as highly specific (100%) for hydatidosis. Therefore purification procedures can strongly affect the diagnostic value of antigens with identical electrophoretic behavior in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).

Animals↗

PubMed1996

Measurement of soluble L-lactate in dairy products using screen-printed sensors in batch mode.

Amperometric sensors based on lactate oxidase and platinized carbon were constructed entirely by screen-printing. They were used to estimate the lactate concentration in diluted samples of yoghurt and buttermilk. Estimates made in untreated (apart from dilution) solutions were subject to bias, relative to values obtained from spectrophotometric assays. Removal of cations improved the accuracy of the estimates. Calcium may have made a small contribution to the bias but the nature of the inhibition is otherwise unknown. With untreated samples, determinations of any accuracy require sufficient dilution to avoid such bias, although the smaller signals resulting from dilution place more demands on calibration and manufacturing standards. The relative standard deviations of estimates, based on variation among sensors, of the most dilute samples were generally too high for practical use. The errors of estimates at lower dilutions approached those of screen-printed sensors made under industrial conditions.

Biosensing Techniques↗

PubMed1996

Inhibition of superoxide dismutase from Ascaris suum by benzimidazoles and synthesized pyrimidine and glycine derivatives.

Copper-zinc superoxide dismutase was purified from Ascaris suum (Nematoda). Four benzimidazole derivatives, six recently synthesized pyrimidine derivatives and eleven recently synthesized glycine derivatives were shown to inhibit: (1) purified extracts of A. suum superoxide dismutase; (2) superoxide dismutase from host liver, and (3) purified extracts of superoxide dismutase from living A. suum incubated in the presence of these drugs. Thiabendazole compounds, with a documented effect against helminth parasites, were found to affect the superoxide dismutase. The inhibitory effects of some pyrimidine and glycine derivatives were higher than those of benzimidazoles, and the pyrimidine compounds failed to inhibit the host's enzyme. These derivatives are candidate anthelmintics, acting as inhibitors of certain metalloenzymes in parasites.

Analysis of Variance↗

PubMed1996

Genetic diversity affects testicular morphology in free-ranging lions (Panthera leo) of the Serengeti Plains and Ngorongoro Crater.

Reduced genetic variability is known to adversely affect ejaculate quality in inbred lions (Panthera leo) physically isolated in the Ngorongoro Crater compared with outbred lions inhabiting the adjacent Serengeti Plains in East Africa. This study compared the histomorphology of testicular biopsies from these two lion populations. Ngorongoro Crater lions had fewer (P < 0.05) seminiferous tubules with spermiogenesis and fewer (P < 0.05) spermatids per seminiferous tubular cross-section than Serengeti Plains lions, although seminiferous tubular diameter did not differ (P > 0.05) between populations. Interstitial areas were greater (P < 0.05) in Crater than in Plains lions, but no qualitative differences were evident, suggesting that proportionately less testicular area was occupied by seminiferous tubules in Crater lions. None of the lions in either population had evidence of testicular degeneration. Overall results suggest that inbred Crater lions have reduced spermiogenesis and less total seminiferous tubular area per testis. These data further support the premise that genetic homogeneity compromises reproductive traits in free-living, male African lions.

Africa↗

PubMed1995

A novel lipid-binding protein from the cestode Moniezia expansa.

A lipid-binding protein (LBP) has been purified from the cytosol of the cestode Moniezia expansa. The native LBP was found to be an oligomer of approx. 250 kDa, consisting of 11 kDa monomers. The LBP bound saturated and unsaturated fatty acids, but not their CoA derivatives, with KD values in the range 0.68-7.8 microM. Cholesterol, dihydroergosterol, bilirubin and retinoids were also bound, but alpha-tocopherol, bile acids, alk-2-enals and alka-2,4-dienals were not. Evidence suggests that there are two binding sites per subunit, each with different specificities. The fluorescent fatty acid 11-[(5-dimethylaminonaphthalene-1-sulphonyl)amino]undecanoic acid (DAUDA) and retinol both showed an additional high-affinity binding site with a density of approximately 0.1 per subunit, suggesting specific binding to the oligomer. The amino acid composition of Moniezia LBP was distinct from that of previously characterized fatty acid-binding proteins (FABPs). The protein was not N-terminally blocked and yielded a unique amino acid sequence, unrelated to that of any known FABP; there was also evidence of microheterogeneity. Polyclonal antibodies raised to the Moniezia protein did not cross-react with mammalian, nematode or digenean FABP. The Gibbs free energy for protein folding (13.02 kJ/mol; 3.1 kcal/mol), determined by urea denaturation, was identical for both the native and ligand-bound Moniezia LBP. CD spectra showed that the Moniezia protein contained 36% alpha-helix and that the secondary structure underwent only minor changes on ligand binding. Moniezia LBP binds a range of anthelmintics, with KD values again in the range 0.66-7.3 microM. It is possible that, in helminths, binding proteins may play a role in determining the specificity and site of action of anthelmintics.

Amino Acid Sequence↗

PubMed1995

Changes in T-cell subpopulations in mice during prolonged experimental secondary infection with Echinococcus granulosus.

Balb/c mice were infected intraperitoneally with protoscoleces of Echinococcus granulosus. After 15 months of infection, and by means of flow cytometry, the expression of T-cell markers CD3, CD4, and CD8 on T cells from peripheral blood, spleen, and thymus was analyzed and compared with that of age-matched controls. Infected mice had higher percentages of CD3+, and CD4+ cells in peripheral blood, and higher percentages of CD8+ cells in the spleen, when compared with control mice. CD4+ and CD8+ cells in peripheral blood and CD8+ cells in thymus also showed higher percentages of expression of interleukin-2 receptor. The results infer a role for interleukin-2 in experimental secondary echinococcosis.

Animals↗

PubMed1995

Influence of hydrogen peroxide on acid-fast staining of Cryptosporidium parvum oocysts.

Infections by the protozoan parasite Cryptosporidium parvum are routinely diagnosed by modified Ziehl-Neelsen (acid-fast) staining of faecal preparations despite the counterstaining and ghost-like appearance of some oocysts. Quantitative studies demonstrated that only a small percentage of oocysts excreted by naturally infected newborn calves displayed acid-fast characteristics, but that percentage increased when the time between excretion and sample staining was increased. The treatment of faecal samples with hydrogen peroxide (10 min, 5 vol. final concentration) caused all oocysts to become acid-fast, with up to 40-fold increases in test sensitivity in samples treated and stained within 3 h of excretion. Flow-cytometry analysis of hydrogen peroxide-treated oocysts also demonstrated increased labelling of oocysts by a commercial monoclonal antibody preparation commonly used for diagnosis.

Acids↗

PubMed1994

Copper-zinc superoxide dismutase from Ascaris suum (Nematoda): purification and characterization.

Copper-zinc superoxide dismutase from Ascaris suum (Nematoda) was purified in a new, more efficient, and faster manner. The process included differential centrifugation, fractionation with ammonium sulfate, and sodium dodecyl sulfate-polyacrylamide electrophoresis, yielding a 340-fold purification (specific activity of 47 units/mg). Optimal storage conditions, optimal pH range, thermostability, molecular weight and ultraviolet-visible absorption spectrum of the enzyme are described, and a new enzymatic model for pharmacological screening is suggested.

Ammonium Sulfate↗

PubMed1993

Dose-dependent effects of hydatid fluid toxins from Echinococcus granulosus on mouse peritoneal macrophages.

Hydatid fluid (HF) from hydatid cysts of the cestode Echinococcus granulosus inhibited the phagocytosis of bacteria and yeast cells by host macrophages in vitro. Different assays were used to study the dose-dependent effect of partially purified HF toxins on peritoneal macrophages (PM phi) of the mouse. Trypan blue exclusion, as well as measuring the specific release of lactate dehydrogenase activity and 51Cr release of 51Cr-labelled PM phi, showed that incubation with HF toxins lead to the lysis of the target cells. By measuring the effect of the toxins on the reduction of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide by PM phi, and by experimental labelling using [3H]-uridine, a decrease in metabolic activity was seen. However, at non-lytic concentrations, the PM phi showed a peak of metabolic activity. The observations of the effects on macrophages by parasite-derived toxins may be related to a mechanism by which the parasite survives within an immunized host.

Animals↗

PubMed1992

Fatal bleeding from duodenal varices as a late complication of neonatal thrombosis of the inferior vena cava.

A 21-year-old man was hospitalized with repeated bleeding from the gastrointestinal tract, the cause of which could not be detected by gastroscopy, colonoscopy or arteriography. After the third surgical intervention he died of uncontrollable bleeding. At autopsy, marked and partly ulcerated duodenal varices were found to be the cause of the bleeding. They were part of a collateral circulation that had developed from chronic obstruction of the inferior vena cava, where thrombosis had arisen in association with neonatal renal vein thrombosis. There was consecutive renal infarction which had required a left nephrectomy on the 2nd day after birth.

Adult↗

PubMed1992

Comparative cytotoxicity of secondary hydatid cysts, protoscoleces, and in vitro developed microcysts of Echinococcus granulosus.

Infection with the metacestode of Echinococcus granulosus is characterized by a concomitant immunity. Survival of established and developing hydatid cysts in the intermediate host implies a mechanism to modulate its immunological reactions. In order to investigate this mechanism, secondary hydatid cysts were isolated from intraperitoneally infected laboratory white mice (strain NMRI) 12 months p.i. A number of hydatid cysts were freed from the surrounding host adventitial tissue. Monolayer cultures of non-stimulated peritoneal macrophages of NMRI mice were prepared and incubated in the presence of the hydatid cysts. By means of a trypan blue exclusion test and by measuring the incorporation of tritium labelled uridine, it was found that the presence of hydatid cysts reduced the viability of the macrophages in vitro. Toxic substances are probably secreted since the medium of cultured hydatid cysts also displayed cytotoxic activity. Hydatid cysts with adventitia, as well as culture medium of those cysts, were less toxic. When toxins, partially purified from hydatid cyst fluid, were previously incubated on a collagen coated surface, a reduced level of toxicity was found, suggesting that collagen of the host adventitia may play a role in controlling the liberation of toxins by the hydatid cyst. Virtually no toxicity was exerted by protoscoleces or by the medium of cultured protoscoleces, in contrast to in vitro vesiculated protoscoleces (so called microcysts). The results reveal a novel feature of hydatid cysts that may play a role in the survival of the parasite in the immunized host.

Animals↗

PubMed1992

A new technique for the rapid dissolution of retained ductal gallstones with monoctanoin in T-tube patients.

Retained gallstones in the biliary ducts have been therapeutically managed with monoctanoin (Moctanin; Ethitek Pharmaceuticals Company, Skokie, IL) since Food and Drug Administration approval in 1985. The clinical usefulness of monoctanoin therapy has previously been regarded by some investigators as limited because of the length of time required to achieve complete dissolution (2 to 10 days) and less than optimal results (50% to 86% efficacy). Here, the authors describe a safe technique for the rapid dissolution of retained stones that they have used successfully in four patients. This technique eliminates the need for pressure monitoring in the T-tube patient and is suitable for short-stay hospitalization. Representative case histories are presented.

Adult↗