Biomedical subjects

Jens Randel Nyengaard

Publications and source records attributed to Jens Randel Nyengaard.

14 recordsLinked to original sources

PubMed2006

Development of overt proteinuria in the Munich Wistar Frömter rat is suppressed by replacement of chromosome 6 in a consomic rat strain.

In a cross between the Munich Wistar Frömter (MWF) rat and spontaneously hypertensive rats (SHR), a major quantitative trait locus (QTL) was identified on rat chromosome 6 (RNO6) that demonstrated the strongest linkage to albuminuria among several QTL identified. The QTL represented the only locus that is linked to both early-onset albuminuria and increased renal interstitial fibrosis in adult animals. A consomic MWF-6(SHR) strain in which chromosome 6 from SHR was introgressed into the MWF background therefore was generated to test the relevance of this QTL. Phenotype analysis at 8 wk of age revealed that early onset of albuminuria in MWF with a 55-fold elevation of urinary albumin excretion compared with SHR (P < 0.0001) was completely abolished in MWF-6(SHR). Time-course analysis until week 24 demonstrated only a moderate increase of urinary albumin excretion in MWF-6(SHR), whereas MWF reached levels in the nephrotic range (16.6 +/- 3.5 versus 162.6 +/- 16.0 mg/24 h; P < 0.0001). At this age, analysis of glomerulosclerosis, tubulointerstitial damage, renal interstitial fibrosis, and renal collagen III mRNA expression revealed a significant improvement of all parameters in MWF-6(SHR) compared with MWF (P < 0.05). At 32 wk, MWF but not MWF-6(SHR) demonstrated overt proteinuria (354.6 +/- 37.6 versus 48.8 +/- 13.2; P < 0.0001), whereas serum urea, cholesterol, and triglyceride concentrations were lower and creatinine clearance was higher in MWF-6(SHR) compared with MWF (P < 0.05). Therefore, although albuminuria in MWF is determined by a complex interplay of several QTL, our data demonstrate that genetic exchange of one locus on RNO6 leads to marked suppression of early-onset albuminuria and renal damage in MWF.

Albuminuria↗

PubMed2006

Glomerular and tubular function during AT1 receptor blockade in pigs with neonatal induced partial ureteropelvic obstruction.

Previously, we showed that neonatal induced chronic partial unilateral ureteral obstruction (PUUO) of the multipapillary pig kidney decreased glomerular filtration rate (GFR) of the obstructed kidney. We hypothesized that ANG II and nitric oxide (NO) are important for the changes in renal function and in the present study we examined the effects of chronic AT1 receptor blockade using CV-11974 (0.12 mg/h candesartan from age 23 to 30 days) on kidney function development after PUUO was induced in 2-day-old piglets. Moreover, the effect of superimposed acute NO inhibition using N(G)-nitro-l-arginine methyl ester (l-NAME; 15 mg/kg) was examined to identify if this has diagnostic potential. PUUO significantly increased GFR in the nonobstructed contralateral kidney independent of candesartan. In candesartan-treated piglets, the l-NAME-induced GFR reduction seen in normal and nonobstructed kidneys was absent in the partial obstructed kidneys. Urine output and fractional excretion of water were increased from the partial obstructed kidneys. Consistent with this immunohistochemical analyses showed a reduced aquaporin-2 labeling in the collecting duct principal cells. Moreover, renal sodium handling was compromised by PUUO evidenced by an increased fractional excretion of sodium which was enhanced by candesartan treatment. In conclusion, our findings suggest that the counterbalance between AT1 receptor-mediated vasoconstriction and NO-mediated vasodilatation which maintain GFR in normal young porcine kidneys is changed by neonatal induced chronic PUUO. This may have diagnostic potential in children with suspected congenital obstruction. Our results also demonstrate compromised tubular functions in response to chronic PUUO despite preservation of glomerular function.

Angiotensin II Type 1 Receptor Blockers↗

PubMed2006

Endothelial proliferation and increased blood-brain barrier permeability in the basal ganglia in a rat model of 3,4-dihydroxyphenyl-L-alanine-induced dyskinesia.

3,4-Dihydroxyphenyl-L-alanine (L-DOPA)-induced dyskinesia is associated with molecular and synaptic plasticity in the basal ganglia, but the occurrence of structural remodeling through cell genesis has not been explored. In this study, rats with 6-hydroxydopamine lesions received injections of the thymidine analog 5-bromo-2'-deoxyuridine (BrdU) concomitantly with L-DOPA for 2 weeks. A large number of BrdU-positive cells were found in the striatum and its output structures (globus pallidus, entopeduncular nucleus, and substantia nigra pars reticulata) in L-DOPA-treated rats that had developed dyskinesia. The vast majority (60-80%) of the newborn cells stained positively for endothelial markers. This endothelial proliferation was associated with an upregulation of immature endothelial markers (nestin) and a downregulation of endothelial barrier antigen on blood vessel walls. In addition, dyskinetic rats exhibited a significant increase in total blood vessel length and a visible extravasation of serum albumin in the two structures in which endothelial proliferation was most pronounced (substantia nigra pars reticulata and entopeduncular nucleus). The present study provides the first evidence of angiogenesis and blood-brain barrier dysfunction in an experimental model of L-DOPA-induced dyskinesia. These microvascular changes are likely to affect the kinetics of L-DOPA entry into the brain, favoring the occurrence of motor complications.

Animals↗

PubMed2006

Does neuroglobin protect neurons from ischemic insult? A quantitative investigation of neuroglobin expression following transient MCAo in spontaneously hypertensive rats.

Neuroglobin (NGB) is a recently characterized heme globin expressed primarily in retinal nerve cells and at very low levels in endocrine-active regions of vertebrate brains. When artificially over-expressed, NGB reduces the infarct size observed after transient Middle Cerebral Artery occlusion (tMCAo) in rats. This study addresses the post-ischemic NGB expression in vivo. Ten Spontaneously Hypertensive Rats (SHRs) were randomized to tMCAo (n = 6) or sham (n = 4), and euthanized 24 h later. NGB mRNA expression was determined by means of quantitative Reverse Transcription Polymerase Reaction (qRT-PCR). Thirteen animals subjected to either 90 min tMCAo (n = 7) or sham (n = 6) surgery, were euthanized 1 week after surgery. Post-ischemic expression of NGB and the neuronal marker NeuN was studied using free-floating immunohistochemistry. Design-based stereological quantification of NGB- and NeuN-positive cells in the striatum was performed using the optical fractionator. Significantly less NGB mRNA was expressed in the ischemic hemispheres of tMCAo animals after 24 h (P < or = 0.002). At the protein level, we found a significantly lower number of NGB- and NeuN-positive striatal neurons in tMCAo rats (P < or = 0.004). NGB expression was mainly confined to the hypothalamus and amygdala. Less than one out of every two thousand neurons expressed NGB in the striatum. In the ischemic territory we did not observe selective sparing of NGB expressing neurons. No significant change in the NGB/NeuN ratio was observed. Our data indicate that endogenous expressed NGB does not provide protection against ischemic injury induced by tMCAo in SHRs.

Amygdala↗

PubMed2006

A stereologic approach to estimate the number of immunogold-labeled molecules in cells of tubules.

OBJECTIVE: The quantity of molecules can be measured very precisely by molecular biological methods, but the capabilities of these are limited to measure only the total mass of tissue. For estimating the number of molecules at the cell level, it is necessary to combine an immunohistochemical protocol with designed-based principles of stereology at the level of electron microscopy (EM). This article focuses on the problems and practical solutions of fitting together immunohistochemistry, stereology, and electron microscopy for the estimation of the number of angiotensin II AT1 receptors in rat kidney arterioles. STUDY DESIGN: We performed the preembedding immunostaining of angiotensin II AT1 receptors using the silver-enhanced immunogold labeling system at EM level on serial sections of renal arterioles from 5 rats. RESULTS: Using this method the number of molecules can be estimated along the renal arterioles separately on the cell's surface, in cytoplasm, in nucleus, or in any subcellular location. CONCLUSION: For estimating the number of AT1 receptors, we designed a protocol that took into account the requirements for both immuno-EM and stereology. This method can be applied for estimating any molecule number in different types of cells in tubules.

Animals↗

PubMed2005

Endotoxemia-induced lymphocyte apoptosis is augmented by a hyperinsulinemic-euglycemic clamp.

BACKGROUND: Sepsis and endotoxemia are associated with lymphocyte apoptosis. This has been regarded as harmful, contributing to further immune suppression in already immune-compromised patients. Because normalization of blood glucose improves outcome in critically ill patients, the authors hypothesized that one of the effects of insulin and normoglycemia would be inhibition of lymphocyte apoptosis. Therefore, in this experimental study in pigs, the authors examined the separate and combined effects of acute endotoxemia and a hyperinsulinemic-euglycemic clamp (HEC) on lymphocyte apoptosis. METHODS: After 60 min of stabilization, 38 anesthetized and mechanically ventilated pigs (weight, 35-40 kg) were divided (by randomization performed before the experiment) into four groups and were then studied for 570 min. Group 1 received no intervention. Group 2 received a HEC (5 mm p-glucose, insulin infusion rate of 0.6 mU . kg (-1). min(-1)) for 570 min. Group 3 received a lipopolysaccharide infusion for 180 min. Group 4 was given a combination of a HEC and a lipopolysaccharide infusion. After the 570-min study period, the pigs were killed, and tissue was sampled from the spleen and frozen. In four sections of each sample, the apoptosis of B and T lymphocytes were analyzed using stereologic methods: The number of apoptotic B and T cells was estimated by fluorescence immunohistochemistry with anti-active caspase-3 and either anti-CD21 (B lymphocytes) or anti-CD3epsilon (T lymphocytes). The number of apoptotic B and T lymphocytes was then compared using two-way analysis of variance, and the interaction between endotoxemia and the clamp (hyperinsulinemia and euglycemia) was investigated. RESULTS: Endotoxemia induced apoptosis of B (P < 0.001) and T lymphocytes (P = 0.016) in the spleen, and this effect was independent of the clamp. The ratios of apoptotic cells in the spleen tissue of pigs with and without endotoxemia were 2.4 (confidence interval, 1.7-3.4) and 1.6 (confidence interval, 1.1-2.2) for B and T lymphocytes, respectively. Independent of endotoxin infusion, HEC increased the number of apoptotic lymphocytes (P = 0.029 and P = 0.038 for B and T lymphocytes, respectively). The ratios of the number of apoptotic spleen cells in pigs treated and not treated with HEC were 1.5 (confidence interval, 1.0-2.1) and 1.5 (confidence interval, 1.0-2.1) for B and T lymphocytes, respectively. CONCLUSION: In this porcine model, both endotoxemia and a HEC increased the number of apoptotic B and T lymphocytes in the spleen. Contrary to our hypothesis, lymphocyte apoptosis during acute endotoxemia was augmented by a HEC.

Algorithms↗

PubMed2004

Myelinated nerve fibres in the subcortical white matter of cerebral hemispheres are preserved in alcoholic subjects.

Previous studies have found that the primary change in the brain of chronic alcoholics is a selective loss of white matter in cerebral hemispheres. It is unknown whether the loss of white matter is due to alcohol-induced degeneration of axons in white matter, and this hypothesis has never been tested. We used a newly developed stereological method to investigate whether the length and diameter of myelinated fibres change in the subcortical white matter of male alcoholic subjects. We found no significant differences between control and alcoholic subjects with respect to the total volume, total length, and mean diameter of myelinated fibres in white matter. Our results do not find any support for the hypothesis that chronic alcoholic subjects may suffer from a degeneration of axons (myelinated fibres) in white matter in cerebral hemispheres.

Alcoholism↗

PubMed2004

Fate of bone formed by guided tissue regeneration with or without grafting of Bio-Oss or Biogran. An experimental study in the rat.

OBJECTIVES: The aim of the study was to examine whether bone produced by guided tissue regeneration (GTR) in combination with Bio-Oss or Biogran is stable on a long-term basis. MATERIAL AND METHODS: Fifty-four, 3-month-old Wistar rats were divided into three groups and rigid, hemispherical, teflon capsules were placed with their open part facing the lateral surface of the exposed mandibular ramus (one capsule per animal). In the first group, the capsules were loosely packed with a standardized quantity of Bio-Oss, in the second group with Biogran, and in the last group were left empty. After 1 year of healing, the capsules were removed. Six animals from each of the 3 experimental groups were killed immediately after capsule removal (baseline), or 3 or 6 months after re-entry. The volume of (1) newly formed bone, (2) remaining biomaterial particles, and (3) soft connective tissue in the space originally created by the capsule was estimated by a point-counting technique in three to four histological sections, taken by uniformly random sampling. RESULTS: While considerable bone formation had occurred in the empty control capsules, only limited bone formation was observed in the two test groups. The major portion of the space originally created by the capsules in the test groups was occupied by biomaterial particles embedded in connective tissue. At baseline, the mean volume of newly formed bone occupied 23% of the original capsule space in the animals grafted with Bio-Oss, 12.6% in those implanted with Biogran, and 94.1% in those that received empty control capsules. Six months after capsule removal, the corresponding values were 21.5%, 13.2%, and 91.7%, respectively. No statistically significant differences were observed between baseline, and the 3-, and 6-month observation times in terms of bone volume for any of the three treatment groups (p>0.05). CONCLUSION: Bone produced by GTR with and without implantation of Bio-Oss, or Biogran, is stable on a long-term basis, but bone formation is obstructed by implantation of these biomaterials.

Animals↗

PubMed2004

Selective loss of calcitonin gene-related Peptide-expressing primary sensory neurons of the a-cell phenotype in early experimental diabetes.

To evaluate the possible role of neuropeptide immunoreactive primary sensory neurons on the development of nociceptive dysfunction in diabetes, the absolute numbers of immunoreactive substance P and calcitonin gene-related peptide (CGRP) dorsal root ganglion (DRG) cell bodies were estimated in diabetic and nondiabetic BALB/C (p75(+/+)) and p75 receptor knockout (p75(-/-)) mice with unilateral sciatic nerve crush. The total numbers of immunoreactive substance P A-cells, substance P B-cells, CGRP A-cells, and CGRP B-cells in L5DRG were estimated using semithick consecutive sections and the optical fractionator. After 4 weeks of streptozotocin-induced diabetes, the number of immunoreactive CGRP A-cells was reduced from 692 +/- 122 to 489 +/- 125 (P = 0.004) in p75(+/+) mice on the noncrushed side. In p75(-/-) mice, there was no such effect of diabetes on the immunoreactive CGRP A-cell number. In p75(+/+) and p75(-/-) mice, there was no effect of diabetes on the immunoreactive CGRP B-cell number, nor was there any effect of diabetes on the immunoreactive substance P B-cell number. Sciatic nerve crush was associated with a substantial loss of L5DRG B-cells in diabetic and nondiabetic p75(+/+) mice and with substantial loss of immunoreactive substance P cells in diabetic p75(+/+) mice. In diabetic and nondiabetic p75(-/-) mice, there was no crush effect on neuropeptide expression. It is concluded that experimental diabetes in the mouse is associated with loss of immunoreactive CGRP primary sensory neurons of the A-cell phenotype, that this loss could play a role for the touch-evoked nociception in the model, and that the neuronal immunoreactive CGRP abnormality possibly is mediated by activation of the p75 neurotrophin receptor.

Animals↗

PubMed2003

Structure and growth of the utricular macula in the inner ear of the slider turtle Trachemys scripta.

In general, postembryonic production of inner ear vestibular hair cells (HCs) is believed to occur in all nonmammalian vertebrates. However, no study on this topic has been published on reptiles and, consequently, it is not known whether this also applies to these vertebrates. Therefore, the present study applied stereological methods in order to estimate the total number of HCs in turtles of varying sizes. The findings are that in prehatchlings the utricular macula (UM) contains approximately 4000 HCs as compared to approximately 5000 in juveniles, approximately 8000 in medium-sized turtles, and approximately 12,000 in large, sexually mature turtles. Scanning electron microscopy (SEM) reveals that presumably newly generated HCs with small surface areas and thin stereovilli are found in all regions of the UM. Furthermore, it reveals that utricular HCs can be classified as belonging to a specific region from the morphology of their apical structure. Striolar HCs have a large free oval-to-ovoid surface, a hair bundle with numerous stereovilli, and a short kinocilium. Rampary and cotillary HCs have smaller and slimmer free surfaces, comparatively fewer stereovilli, but much longer kinocilia. In conclusion, the current study demonstrates that postembryonic production of HCs does occur in reptiles and thereby supports the general view that this is a common trait in all nonmammalian vertebrates.

Age Factors↗

PubMed2003

Deproteinized bovine bone (Bio-Oss) and bioactive glass (Biogran) arrest bone formation when used as an adjunct to guided tissue regeneration (GTR): an experimental study in the rat.

OBJECTIVES: Grafting of deproteinized bovine bone or bioactive glass has been suggested as an adjunct to guided tissue regeneration (GTR) for the treatment of periodontal and peri-implant bone defects but the influence of these materials on bone formation is not clarified. The aim of the study was to examine the long-term influence of deproteinized bovine bone (Bio-Oss) or bioactive glass (Biogran) on bone formation produced by GTR. MATERIAL AND METHODS: Eighteen rats were used. Following incisions along the inferior border of the mandible, muscle-periosteal flaps were raised to expose the mandibular ramus. Rigid, hemispherical, Teflon capsules (6 mm internal diameter and 1 mm peripheral collar) loosely packed with a standardized quantity of either deproteinized bovine bone (test group 1) or bioactive glass (test group 2), or empty capsules (control group) were then placed with their open part facing the lateral surface of the ramus (one capsule per animal). After 1 year, the capsules were removed by a reentry operation, and the animals were sacrificed. Histological specimens of the augmented sites were prepared, and the volumes of (1). newly formed bone, (2). graft particles, and (3). soft connective tissue in the space originally created by the capsule were estimated by a point-counting technique in three to four histological sections, taken by uniformly random sampling. RESULTS: Limited bone formation was observed in the two test groups. The major part of the space originally created by the capsules was occupied by graft particles embedded in connective tissue. The mean volume of newly formed bone occupied only 23% of the total space in the animals grafted with Bio-Oss and 12.6% in those grafted with Biogran. In the control animals, however, 88.2% (p<0.01) of the space was filled with newly formed bone. There were no signs of ongoing bone formation in any of the three experimental groups. CONCLUSION: It is concluded that grafting of Bio-Oss or Biogran as an adjunct to GTR arrests bone formation.

Animals↗

PubMed2003

TGF-beta1 induces aberrant laminin chain and collagen type IV isotype expression in the glomerular basement membrane.

Transforming growth factor-beta1 (TGF-beta1) contributes to the thickening of the glomerular basement membrane (GBM), abnormal deposition of extracellular matrix (ECM) therein and expansion of the mesangial matrix (MM) in several glomerular kidney diseases. However, the influence of TGF-beta1 on the expression of collagen IV isotypes and laminin chains in the GBM and the MM in vivo is not known in detail. By using transgenic mice with TGF-beta1 expression targeted to the juxtaglomerular apparatus and a combination of immunohistochemistry, Western blotting, immunoelectron microscopy and in situ hybridization, we investigated the contribution of different laminin chains and collagen type IV isotypes to the basement membrane thickening and mesangial expansion. We report that exposure of the glomerulus to TGF-beta1 in vivo induces aberrant deposition of fetal laminin alpha1, alpha2 and beta1 chains and collagen type IValpha1/alpha2 in the GBM. On the other hand, the TGF-beta1-mediated expansion of the mesangial ECM is dominated by the normal components. We found that the cellular origin of at least laminin alpha1 and alpha2 chains may be the glomerular endothelial cells. We speculate that the endothelial cells could contribute to TGF-beta1-induced glomerulopathy and should be considered as target cells for early intervention in glomerular diseases associated with TGF-beta1 in man.

Animals↗

PubMed2002

Measurements of cortical interstitium in biopsies from human kidney grafts: how representative and how reproducible?

BACKGROUND: The aim of the study was to evaluate how well a biopsy from one region of a human renal allograft represented biopsies from other regions regarding the renal interstitial tissue assessed by stereology. Furthermore, we wanted to evaluate the reproducibility of the measurements. METHODS: Punch biopsies (3 mm) from six regions in each kidney were obtained from seven explanted renal grafts with varying degrees of clinically diagnosed chronic and acute rejection. One kidney, removed for a minor pelvic tumour, served as reference material. Using point counting on PAS-stained sections, the volume fraction of the interstitial tissue per glomerular cortex V(V)(interstitium/cortex) was estimated. From each kidney, two of the six biopsies were re-evaluated by the same observer. RESULTS: V(V)(interstitium/cortex) varied from 0.25 to 0.78 between the explanted kidneys vs 0.26 in the reference kidney. Variations within the kidneys were low, expressed by standard deviations (SD) of between 0.04 and 0.06, and coefficients of variation (CV) between 0.06 and 0.22. The SD estimated from repeated measurements was 0.04 and CV was 0.07. CONCLUSIONS: Biopsies from one region of the kidney were found to be representative for estimates of interstitial tissue in explanted human kidney grafts, and the degree of reproducibility was high when using point counting, as in the present study.

Adult↗

PubMed2002

Reduced number of glomeruli in kidneys with neonatally induced partial ureteropelvic obstruction in pigs.

PURPOSE: We quantify the structural components of the nephron in adult pig kidneys with neonatally induced unilateral hydronephrosis in comparison with nonobstructed kidneys. MATERIALS AND METHODS: The study included 11 pigs with unilateral partial ureteropelvic obstruction induced 2 days after birth and 8 sham operated control pigs. Obstructed kidney glomerular filtration rate was significantly reduced at age 4 weeks but did not differ from control kidneys after 24 weeks. At age 24 weeks the kidneys were perfusion fixed, and the number and volume of glomeruli and tubular lengths were measured using stereological methods. RESULTS: Mean obstructed kidney volume did not differ from that of control kidneys. Mean number plus or minus standard deviation of glomeruli in the obstructed kidneys was reduced by 28% compared to that of control kidneys (502 +/- 163 x 103 versus 697 +/- 161 x 103, p = 0.02), whereas no difference in mean glomerular volume was observed. Mean length of the proximal or distal tubules did not differ between obstructed and control kidneys. Mean number or volume of glomeruli in nonobstructed kidneys contralateral to obstructed kidneys did not differ from that of control kidneys. The individual number of glomeruli in the obstructed kidneys was not associated with function of these kidneys. CONCLUSIONS: Neonatally induced unilateral partial ureteropelvic obstruction causes impaired nephrogenesis with a significant reduction in the number of nephrons, which is not reflected in measurements of kidney function in this model. The reduction in the number of glomeruli suggests that congenital unilateral obstruction impairs nephrogenesis.

Animals↗