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Biomedical subjects

J Bakker

Publications and source records attributed to J Bakker.

At least 73 records · Page 4Linked to original sources

Changes in mediobasal hypothalamic gonadotropin-releasing hormone messenger ribonucleic acid levels induced by mating or ovariectomy in a reflex ovulator, the ferret.

The ferret is a reflex-ovulating species in which receipt of an intromission induces a prolonged (+/- 12 h) preovulatory LH surge in the estrous female. This LH surge is probably stimulated by a large release of GnRH from the mediobasal hypothalamus (MBH). In Exp 1 we asked whether GnRH messenger RNA (mRNA) levels increase in response to mating so as to replenish the MBH GnRH stores needed to sustain the preovulatory LH surge. Estrous females were killed 0, 0.25, 0.5, 1, 3, 6, 14, or 24 h after the onset of a 10-min intromission from a male. Coronal brain sections ranging from the rostral preoptic area caudally to the posterior hypothalamus were processed for in situ hybridization using a 35S-labeled oligoprobe complementary to the human GnRH-coding region. We found no evidence of increased MBH GnRH mRNA levels during the ferret's mating-induced preovulatory LH surge. Instead, the number of GnRH mRNA-expressing cells dropped significantly in the arcuate region beginning 6 h after onset of intromission and remained low thereafter. Furthermore, cellular GnRH mRNA levels decreased in the arcuate region toward the end of the preovulatory LH surge. In Exp 2 we asked whether ovarian hormones regulate MBH GnRH mRNA levels in the female ferret. Ovariectomy of estrous females significantly reduced the number of GnRH mRNA-expressing cells in the arcuate region. This decrease was probably not due to the absence of circulating estradiol. Gonadally intact anestrous females had levels of MBH GnRH mRNA similar to those in estrous females even though plasma estradiol levels were equally low in anestrous females and ovariectomized females. Ovarian hormones other than estradiol may stimulate MBH GnRH mRNA levels in anestrous and estrous females.

Animals↗

Evaluation of electrical impedance tomography in the measurement of PEEP-induced changes in lung volume.

STUDY OBJECTIVES: A new noninvasive practical technique called electrical impedance tomography (EIT) was examined for the measurement of alveolar recruitment. DESIGN: Prospective clinical study. SETTING: ICU of a general hospital. PATIENTS: Acute respiratory failure (ARF) patients. MEASUREMENTS: The ventilation-induced impedance changes (VICs) of the nondependent and the dependent part of the lung were determined by EIT as a measure of tidal volume distribution. By the use of an impedance ratio (IR), defined as the VIC of the nondependent part of the lung divided by the VIC of the dependent part of the lung, the ventilation performances in both parts of the lung were compared to each other. RESULTS: Between patients, the VIC of the nondependent part of the lung was significantly lower in the patients with a level of positive end-expiratory pressure (PEEP) of > 10 cm H2O than in patients with a PEEP of < 5 cm H2O (p < 0.05). A significantly lower IR (-/+ SD) was found in the group with PEEP of > 10 cm H2O than in the group with PEEP between 0 and 5 cm H2O (1.28+/-0.58 vs 2.99+/-1.24, respectively; p < 0.01). In individual patients, the VIC of the whole lung increased when the PEEP level was increased. The VICs of the nondependent part of the lung and of the dependent part of the lung showed significant increases at a PEEP of 10 cm H2O compared to a PEEP of 0 cm H2O (p < 0.05). Also the IR decreased in individual patients when the PEEP was increased; a significant decrease was found at 10 cm H2O compared to 0 cm H2O (1.67+/-1.24 vs 2.23+/-1.47, respectively; p < 0.05). CONCLUSIONS: The decrease in IR indicates an increase in VIC in the dependent part of the lung above the nondependent part of the lung. The increase in VIC can be regarded as an increase in lung volume, implying alveolar recruitment in the dependent part of the lung. The same results also have been shown in earlier reports by CT scan. Since EIT is far more practical than CT scanning and also is a bedside method, EIT might help in the adjustment of ventilator settings in ARF patients.

Acute Disease↗

Electrical impedance tomography in the assessment of extravascular lung water in noncardiogenic acute respiratory failure.

STUDY OBJECTIVES: To establish the value of electrical impedance tomography (EIT) in assessing pulmonary edema in noncardiogenic acute respiratory failure (ARF), as compared to the thermal dye double indicator dilution technique (TDD). DESIGN: Prospective clinical study. SETTING: ICU of a general hospital. PATIENTS: Fourteen ARF patients. INTERVENTIONS: In order to use the TDD to determine the amount of extravascular lung water (EVLW), a fiberoptic catheter was placed in the femoral artery. MEASUREMENTS AND MAIN RESULTS: Fourteen consecutive ARF patients receiving mechanical ventilation were measured by EIT and TDD. EIT visualizes the impedance changes caused by the ventilation in two-dimensional image planes. An impedance ratio (IR) of the ventilation-induced impedance changes of a posterior and an anterior part of the lungs was used to indicate the amount of EVLW. For the 29 measurements in 14 patients, a significant correlation between EIT and TDD (r = 0. 85; p < 0.001) was found. The EIT reproducibility was good. The diagnostic value of the method was tested by receiver operator characteristic analysis, with 10 mL/kg of EVLW considered as the upper limit of normal. At a cutoff level of the IR of 0.64, the IR had a sensitivity of 93%, a specificity of 87%, and a positive predictive value of 87% for a supranormal amount of EVLW. Follow-up measurements were performed in 11 patients. A significant correlation was found between the changes in EVLW measured with EIT and TDD (r = 0.85; p < 0.005). CONCLUSION: We conclude that EIT is a noninvasive technique for reasonably estimating the amount of EVLW in noncardiogenic ARF.

Acute Disease↗

Cloning of a trans-spliced glyceraldehyde-3-phosphate-dehydrogenase gene from the potato cyst nematode Globodera rostochiensis and expression of its putative promoter region in Caenorhabditis elegans.

Reverse genetics to determine the relative importance of individual pathogenicity factors of the potato cyst nematode Globodera rostochiensis depends, apart from an efficient transformation protocol for this obligatory plant parasite, on the availability of an efficient promoter. PCR-based cloning was used to isolate a cDNA encoding glyceraldehyde-3-phosphate-dehydrogenase (GAPDH, a crucial enzyme in glycolysis and gluconeogenesis; this gene was designated gpd) and its 5'-flanking region. The cDNA includes 1047 nucleotides encoding an open reading frame that shows high homology with GAPDHs from Caenorhabditis elegans and other species. Analysis of the 745 bp 5'-flanking region of the gpd gene showed no homology with a similar region in C. elegans. In this region several eukaryotic promoter elements are present. 5' Rapid amplification of cDNA ends revealed this gene was trans-spliced with a SL1 spliced leader. The 5'-flanking region of the gpd gene was fused to green fluorescent protein reporter gene and microinjected into the gonads of C. elegans. Green fluorescent protein expression, under the transcriptional control of the 5'-flanking region of gpd, was mainly observed in body wall muscles of transgenic animals. This putative promoter region of GAPDH could be a valuable tool to drive gene expression in transgenic G. rostochiensis and other related plant-parasitic nematode species.

Amino Acid Sequence↗

Effects of liposome-encapsulated drugs on macrophages: comparative activity of the diamidine 4',6-diamidino-2-phenylindole and the phenanthridinium salts ethidium bromide and propidium iodide.

Liposomes can be used for the intracellular delivery of drugs into macrophages. Previously, we developed a liposome-mediated macrophage 'suicide' technique based on the intraphagocytic accumulation of the liposomally delivered bisphosphonate clodronate. Later we found that the diamidine propamidine is even more effective in this approach. In the present study it is shown that liposome-encapsulated 4', 6-diamidino-2-phenylindole (L-DAPI), another well known DNA-binding diamidine, is the most effective drug in killing liver macrophages (Kupffer cells), when intravenously administered in rat. Compared to liposome-encapsulated propamidine (L-propamidine) it showed about 10-fold more activity on a molar basis. Furthermore, L-DAPI was found to induce cell death by inducing apoptosis. The structurally strongly related phenanthridinium salts ethidium bromide (EB) and propidium iodide (PI) exert marked differences in their efficacy. Whereas liposome-encapsulated PI (L-PI) was about 5 times more active in killing macrophages than L-propamidine, liposome-encapsulated EB (L-EB) showed a strongly reduced activity (10 times less than L-PI). As is shown here, PI remains mainly encapsulated in liposomes, while substantial amounts of EB leak out of liposomes. This may very well explain the differences in in vivo activity between L-EB and L-PI.

Animals↗

Endogenous cellulases in animals: isolation of beta-1, 4-endoglucanase genes from two species of plant-parasitic cyst nematodes.

beta-1,4-Endoglucanases (EGases, EC 3.2.1.4) degrade polysaccharides possessing beta-1,4-glucan backbones such as cellulose and xyloglucan and have been found among extremely variegated taxonomic groups. Although many animal species depend on cellulose as their main energy source, most omnivores and herbivores are unable to produce EGases endogenously. So far, all previously identified EGase genes involved in the digestive system of animals originate from symbiotic microorganisms. Here we report on the synthesis of EGases in the esophageal glands of the cyst nematodes Globodera rostochiensis and Heterodera glycines. From each of the nematode species, two cDNAs were characterized and hydrophobic cluster analysis revealed that the four catalytic domains belong to family 5 of the glycosyl hydrolases (EC 3.2.1, 3.2.2, and 3.2.3). These domains show 37-44% overall amino acid identity with EGases from the bacteria Erwinia chrysanthemi, Clostridium acetobutylicum, and Bacillus subtilis. One EGase with a bacterial type of cellulose-binding domain was identified for each nematode species. The leucine-rich hydrophobic core of the signal peptide and the presence of a polyadenylated 3' end precluded the EGases from being of bacterial origin. Cyst nematodes are obligatory plant parasites and the identified EGases presumably facilitate the intracellular migration through plant roots by partial cell wall degradation.

Amino Acid Sequence↗

Estimation of non-cardiogenic pulmonary oedema using dual-frequency electrical impedance.

The study investigates the effects of non-cardiogenic oedema, especially the accumulation of protein in extracellular fluid, on thoracic impedance and proposes a new method of oedema measurement based on an impedance ratio from a dual-frequency measurement. In vitro measurements in a cell containing an albumin-in-saline solution yield a resistance increase when the albumin concentration increases. Subsequently, 13 patients having acute respiratory failure are measured. The single-frequency Z0 measurements and the proposed impedance ratio are compared with extravascular lung water (EVLW) determined by the double indicator dilution method. The single-frequency measurement correlates poorly with EVLW (r = -0.24, p = 0.56). In some patients, a total thoracic impedance increase is found with increasing EVLW. The correlation between the impedance ratio and EVLW is r = -0.79 (p < 0.0005). The ratio decreases as EVLW increases. Thus, when oedema is measured using bio-impedance, cardiogenic and non-cardiogenic oedema yield different results. It is well recognised that cardiogenic oedema decreases total thoracic impedance. In non-cardiogenic oedema, however, protein accumulation causes an impedance increase. The decrease in the impedance ratio as EVLW increases can be explained by the accumulation of albumin in the extracellular compartment.

Adult↗

In vitro measurement of kidney size: comparison of ultrasonography and MRI.

In this in vitro study, the accuracy and repeatability of magnetic resonance imaging (MRI) and ultrasound (US) in assessing renal length and volume were determined. US and MR images of 20 cadaver pig kidneys were obtained twice and evaluated by two observers for each modality. The fluid displacement method provided the "gold standard." Renal volumes were calculated from the US and MR images using the ellipsoid formula. Additional volume calculations after segmentation of the kidney on MR images were done using the voxel-count method. Volumes calculated with the ellipsoid formula resulted in an average of 24% underestimation (range 5%-48%) of the renal volume for both US and MRI. With the voxel-count method, no significant deviation from the true renal volume was encountered. Repeatability was also greatest with the voxel-count method. Measuring renal length, repeatability was, again, better with MRI compared to US. For reliable calculation of renal size in vitro, MRI with use of the voxel-count method is preferred.

Animals↗

The influence of extravascular lung water on cardiac output measurements using thoracic impedance cardiography.

The purpose of this study was to investigate the influence of pulmonary oedema as measured with the double indicator dilution technique on the accuracy of cardiac output (CO) measurement using thoracic impedance cardiography (TIC) compared with thermodilution in thirteen sepsis patients. Differences in the Kubicek and Sramek-Bernstein equation with respect to pulmonary oedema were explored theoretically and experimentally. From a parallel two cylinder model a hypothesis can be derived that CO determined with the Kubicek equation is oedema independent, whereas CO determined using the Sramek-Bernstein equation is oedema dependent. Experimentally, CO determined using Kubicek's equation correlated better with thermodilution CO (r = 0.75) than CO determined with the Sramek-Bernstein equation (r = 0.25). The effect of oedema on the accuracy of TIC was investigated by comparing the differences in the CO of impedance and thermodilution to the extravascular lung water index. For the Kubicek equation the difference was not influenced by oedema (r = 0.04, p = 0.84), whereas for the Sramek-Bernstein equation the difference was affected by oedema (r = 0.39, p = 0.05). Thus, the effects of pulmonary oedema on the accuracy of TIC measurements can better be understood with the parallel cylinder model. Moreover, the Kubicek equation still holds when pulmonary oedema is present, in contrast to the Sramek-Bernstein equation.

Adult↗

Renal artery stenosis and accessory renal arteries: accuracy of detection and visualization with gadolinium-enhanced breath-hold MR angiography.

PURPOSE: To determine the accuracy of gadolinium-enhanced breath-hold magnetic resonance (MR) angiography in the diagnosis of renal artery stenosis and visualization of accessory renal arteries. MATERIALS AND METHODS: Forty-four patients suspected of having renal artery stenosis and 10 potential kidney donors, all of whom were scheduled to undergo elective intraarterial digital subtraction angiography (DSA), were studied. Three-dimensional gradient-echo gadolinium-enhanced MR angiography was performed at 1.5 T with the following parameters: repetition time, 13.5 msec; echo time, 3.5 msec; flip angle, 60 degrees; 195 x 512 matrix; 400-mm field of view; and 6-cm imaging volume consisting of 15 4-mm-thick partitions reconstructed every 2 mm. Gadopentetate dimeglumine (30 mL) was injected with a power injector. MR angiograms were assessed before the standard of reference, intraarterial DSA, was performed. RESULTS: Four MR angiograms were not evaluable because of poor image quality. MR angiography enabled visualization of all but one of the 121 arteries. In four small accessory arteries, a stenosis could not be excluded owing to inadequate spatial resolution. MR angiography enabled the correct diagnosis in 30 of the 31 arteries with a grade 2 (50%-99%) stenosis and in seven of the 10 occluded arteries. Sensitivity and specificity for correct identification of a grade 2 stenosis were 97% and 92%, respectively. CONCLUSION: Gadolinium-enhanced MR angiography is an accurate, minimally invasive method for detecting renal artery stenosis and is reliable for visualizing accessory renal arteries.

Adult↗

Improving scFv antibody expression levels in the plant cytosol.

Expression of single-chain antibody fragments (scFvs) in the plant cytosol is often cumbersome. It was unexpectedly shown that addition at the C-terminus of the ER retention signal KDEL resulted in significantly improved expression levels. In this report the cytosolic location of the scFv-CK was confirmed, excluding possible mistranslocation to other subcellular compartments. It was shown that expression of several other scFvs was also improved in tobacco protoplasts. In addition expression was improved in transgenic potato. Changing from KDEL to KDEI did not affect the enhanced protein expression level. Addition of the KDEL motif is a simple and straightforward tool to stabilize in planta cytosolic expression of many scFvs.

Carboxylic Ester Hydrolases↗

Use of allele specificity of comigrating AFLP markers to align genetic maps from different potato genotypes.

The allele specificity of AFLP markers was assessed in five relatively unrelated potato genotypes. To this end, two diploid mapping populations of potato, F1SH x RH and F1AM x RH, were analysed using four and six AFLP primer combinations, respectively, recently applied to the analysis of the genetically well characterized backcross population BC_C x E. The AFLP profiles of the five parents revealed 733 AFLP markers and, when identical primer combinations were used, 131 comigrating AFLP markers were identified. After construction of five parental maps, the genomic positions of these comigrating AFLP markers were compared and 117 markers (89%) which targeted the same genomic region were assumed to be homologous. Of these putative homologues, 20 markers, each cloned from at least two genotypes, were sequenced and 19 sets of amplification products were shown to be nearly identical. The number of AFLP markers previously mapped in population BC_C x E ranged from three to eleven per chromosome, which allowed a reliable assessment of chromosome numbers from individual linkage groups obtained in populations F1SH x RH and F1AM x RH. The high incidence of corresponding AFLP alleles was confirmed by using an additional set of five primer combinations. The 733 AFLP markers localized provide a valuable reference collection for future mapping studies in potato. As a consequence AFLP analysis may replace more laborious locus-specific marker techniques.

Alleles↗

Transient suppression of macrophage functions by liposome-encapsulated drugs.

Macrophages play an important role in host defense reactions, for example, by phagocytosis of particulate materials. This process also results in the rapid removal of targeting devices such as liposomes and adenovirus vectors and of non-autologous grafted cells and materials. Another aspect of macrophage function is their production and secretion of proinflammatory cytokines. Transient and organ-specific suppression of macrophage function by liposome-mediated manipulation has been shown to improve the efficacy of drug and gene targeting and to reduce the symptoms of inflammatory reactions.

Animals↗

HLA gene haplotype frequencies in bone marrow donors worldwide registries.

To calculate reliable HLA gene and haplotype frequencies of bone marrow donors in various regions in the world, we have analyzed the HLA-A, -B, and -DR phenotype frequencies of 18 bone marrow donor registries with a total of more than 300,000 HLA-A, -B-typed donors. These registries were included in the 22nd edition of Bone Marrow Donors Worldwide. Maximum likelihood gene frequencies, Hardy-Weinberg equilibrium fit, and 2- and 3-locus haplotype frequencies were calculated as well as deltas, relative deltas, and their significance. Remarkable gene and haplotype frequency differences exist between the registries. The genetic distances between the different registries were used to draw phylogenetic trees that clearly show that the degree of similarity between registries is related to their geographic locations. The resulting frequencies can be used for the estimation of the probability of finding a hyplotypically identical related or unrelated bone marrow donor for an individual patient. Phylogenetic trees are useful representations of the similarity between donor pools and can also aid in the selection of donors.

Bone Marrow Transplantation↗