Effects of different perfusates on functional parameters of isolated perfused dog kidneys.

BACKGROUND: The isolated perfused canine kidney has been established as a valid model for conducting both renal physiology and transplantation research. This model is of particular importance for developing new strategies to improve graft function after renal transplantation. In the present study, a newly developed method using isolated haemoperfused porcine kidneys was adapted for use in canine kidneys. In contrast to haemoperfusion, synthetic perfusion media can be standardized and can prevent the initiation of blood-mediated reperfusion reactions. Thus, an additional aim was to determine whether blood could be replaced by synthetic cell-free perfusion solutions. METHODS: Canine kidneys (n = 30) were harvested from donors euthanized in veterinary practices for causes unrelated to the present study. The kidneys were isolated and perfused with autologous blood or cell-free synthetic electrolyte buffer (Tyrode solution). During perfusion, we monitored renal perfusate flow (RPF), glomerular filtration rate (GFR), electrolyte and glucose reabsorption, oxygen consumption and urine concentration. RESULTS: Changes in perfusion medium did not affect the RPF. In contrast, GFR, urine concentration and oxygen consumption were significantly higher, whereas fractional excretion of sodium and glucose were significantly lower in blood- than in Tyrode-perfused kidneys. CONCLUSIONS: This system offers a simple model for studying whole-organ functional alterations after acute renal ischaemia. Renal function indicators were below values reported during in vivo physiological conditions. These functions were better conserved when kidneys were perfused with autologous blood than with Tyrode.     

Molecular cytogenetic characterization of Thinopyrum and wheat--Thinopyrum translocated chromosomes in a wheat--Thinopyrum amphiploid

The wheat--Thinopyrum amphiploid 'Agrotriticum # 3425' (AT 3425), which is highly resistant to Cephalosporium stripe, was identified to carry seven pairs of Thinopyrum chromosomes, three pairs of wheat--Thinopyrum translocated chromosomes and 18 pairs of wheat chromosomes. Fluorescence genomic in situ hybridization (FGISH), C-banding, sequential C-banding and FGISH, and denaturing polyacrylamide gel electrophoresis (SDS-PAGE) were used to characterize and identify the chromosomes. The Thinopyrum chromosomes in AT 3425 were designated as T1 through T7 based on their C-banding patterns. The FGISH and C-banding patterns of mitotic chromosomes in AT 3425 and meiotic chromosomes in the hybrid between AT 3425 and wheat cultivar 'Chinese Spring' (CS) revealed that wheat chromosomes 1D, 2B and 3D were involved in the three wheat-Thinopyrum chromosome translocations designated as (W-T)1, (W-T)2, and (W-T)3 respectively. The analysis of high-molecular-weight glutenin subunits in single seeds of AT 3425 confirmed the involvement of wheat chromosome 1D in the translocation (W-T)1. The designations 1DSuu.1DL-1TL, 2BSuu.2BL-2TL and 3DSuu.3DL-3TL were suggested for the wheat--Thinopyrum translocated chromosomes (W-T)1, (W-T)2 and (W-T)3 in AT 3425 respectively.     

Comparative Analysis of Crossover Exchanges and Chiasmata in Allium Cepa × Fistulosum After Genomic In Situ Hybridization (GISH)

Genomic in situ hybridization (GISH) successfully differentiated homoeologous genomes in the interspecific hybrid Allium cepa × fistulosum, thus allowing the detection of reciprocal crossover events as label exchanges in separating anaphase I chromosomes. Three of the eight chromosome pairs were positively identified by fluorescence in situ hybridization (FISH) to rDNA sequences. There was a general similarity of the GISH-based label exchange frequencies and metaphase I chiasma frequencies, but with a 20% deficit of chiasmata. Reasons for this apparent deficit are discussed. The locations of chiasmata and label exchanges are in broad agreement.     

交感神经节培养的形态学观察

目的：观察交感神经节在体外培养中的生长和形态学特点。方法：无菌，取新生大鼠椎旁交感神经节，用Ｍａｘｉｍｏｗ培养法在相差显微镜下动态观察７天。终止２后，用Ｈｏｌｍｅｓ还原银染色法显示神经纤维。结果：交感神经节在壁后１２－１８ｈ开始生长，２４－４８ｈ由交感神经节长出的纤维长约０．５ｍｍ，７２－９６ｈ可见神经纤维伸长到１－２ｍｍ以上，并呈放射状向四周延伸。Ｈｏｌｍｅｓ还原银染色的标本中可见大量银染的神经     

Relationship between cellular ATP content and cellular functions of primary cultured rat hepatocytes in hypoxia

The importance of oxygen in maintaining the functional integrity of hepatocytes has been well established in a variety of experimental models, such as in vivo , perfused liver and isolated hepatocytes. However, one of the shortcomings of these systems is their short life span. Therefore, we have examined the effects of long-term hypoxia on cellular adenine nucleotide content and cellular functions, such as albumin production, urea production and DNA synthesis, in adult rat hepatocytes in primary culture. Hepatocytes were cultured at a density of 11 × 10⁴ and 5 × 10⁴ cells/0.18 mL per cm² for the study of albumin and urea production and DNA synthesis, respectively, at various oxygen tensions (20, 12, 8 and 5%) for 24 h. Cellular ATP content in cultured hepatocytes in hypoxia gradually declined, corresponding to the decrease in oxygen tension, and the cellular ATP level at 5% oxygen was approximately 20% of that at 20% oxygen. Albumin production also decreased in parallel with the decrease in cellular ATP content in cultured hepatocytes in hypoxia. However, even when cellular ATP content gradually declined corresponding with the decrease in oxygen tension in cultured hepatocytes in hypoxia, such as at 8 or 5% oxygen, urea production remained at a high level; in contrast, DNA synthesis was completely suppressed. These results suggest that the cellular ATP content decreases in cultured hepatocytes during long-term hypoxia in relation to oxygen tension and that the relationship between decreased ATP levels and liver function in cultured hepatocytes during hypoxia differs for albumin production, urea production and DNA synthesis.     

The efficacy of density gradients for islet purification: a comparison of seven density gradients

Seven different density gradient-forming materials were compared as to their efficacy for rat islet purification. Continuous density gradients were used in order to determine the bouant densities of the different pancreatic tissue components. Hand-picked islets served as a control. A significant separation of large numbers of slets from the exocrine tissue band was only seen in the albumin, dextran-40, and metrizamide gradients. Pure islet preparations could not be obtained with any of the gradients studied as none of the gradients completely separated lymph nodes, vessels, and ducts from the islets. Gradients containing sucrose resulted in low islet yields. The islet yields obtained with the other gradients were in the same range as those obtained by hand-picking. Metrizamide gave significantly higher yields than the widely used Ficoll. Judged both in terms of numbers of islets and their in vitro function, the best results were obtained with metrizamide and dextran-40.     

Glial cell line-derived neurotrophic factor (GDNF) gene expression in the human brain: A post mortem in situ hybridization study with special reference to Parkinson's disease

Summary Glial cell line-derived neurotrophic factor (GDNF) is a potent neurotrophic factor for dopaminergic neurons. Since dopaminergic neurons degenerate in Parkinson's disease, this factor is a potential therapeutical tool that may save dopaminergic neurons during the pathological process. Moreover, a reduced GDNF expression may be involved in the pathophysiology of the disease. In this study, we tested whether altered GDNF production may participate in the mechanism of cell death in this disease. GDNF gene expression was analyzed by in situ hybridization using riboprobes corresponding to a sequence of the exon 2 human GDNF gene. Experiments were performed on tissue sections of the mesencephalon and the striatum from 8 patients with Parkinson's disease and 6 control subjects matched for age at death and for post mortem delay. No labelling was observed in either group of patients. This absence of detectable expression could not be attributed to methodological problems as a positive staining was observed using the same probes for sections of astroglioma biopsies from human adults and for sections of a newborn infant brain obtained at post-mortem. These data suggest that GDNF is probably expressed at a very low level in the adult human brain and its involvement in the pathophysiology of Parkinson's disease remains to be demonstrated. GDNF may represent a powerful new therapeutic agent for Parkinson's disease, however.     

Laser Doppler flowmetry of blood perfusion in mucoperiosteal flaps covering membranes in bone augmentation and implant procedures. A pilot study in dogs.

In guided tissue regeneration (GTR) procedures, flap recession or sloughing may occur as an unwanted sequel to the placement of a membrane. This study was designed to assess the applicability of laser Doppler flowmetry (LDF) in the evaluation of blood perfusion in the mucoperiosteal flap covering the membrane. Five Labrador dogs were initially used inthe study, but one animal was later excluded due to post‐operative problems. Maxillary premolar teeth were extracted and full thickness mucoperiosteal flaps were raised. Following removal of the buccal bone plate, 4 titanium implants were placed on each side. An experimental biodegradable polylactic‐acid membrane was placed over the fixtures on one side to allow for GTR. The mucoperiosteal flap was repositioned and secured with sutures. The contralateral side served as control with no membrane. Blood perfusion was measured in the flaps before surgery, immediately after suturing and at 24, 48 and 72 h postoperatively. A laser Doppler flowmeter was used to assess the blood perfusion. In 3 animals the membrane was exposed within 2 weeks post‐operatively, and in these 3 animals the LDF values were lower on the membrane side than on the control side. The mean LDF value was lower on the membrane side for each of the 4 periods studied. The tindings suggest that LDF can be a valuable method to study blood perfusion of oral mucosal flaps and that there may be a relationship between a reduced relative LDF value and subsequent exposure of the membrane to the oral environment.     

Surgical correction of convergent strabismus: its relationship to prism compensation

Esotropic patients whose angle of strabismus has been corrected by prisms frequently increase their angle deviation to compensate for the prismatic correction. This sensorio-motorial reaction to prism correction has been given the name of anomalous movements (a.m.). Quantification of a.m. has been made according to the amount of prisms that an esotropic patient is capable of compensating for (progressive prism compensation test - p.p. test). Some esodeviation does not compensate for any prisms at all since a.m. have not yet developed. Other cases compensate for as much as 40 or 60 prism diopters and more of over-correction of the angle deviation and they therefore have powerful a.m. The interference of these innervational forces acting on the medial recti to corrective surgery has been studied in 126 operated esotropic patients. A significant decrease from the expected surgical result (p < 0.001) has been found in patients having powerful a.m., as can be judged by the p.p. test. It is believed that a.m. are an important drawback contributing to vitiate any formula on the amount of muscle surgery to be performed in patients having no possibilities of restoring normal binocular vision.Practical advice on how to eliminate this drawback and theoretical reasoning on the significance of a.m. are offered.     

Ocular perfusion pressures in different types of glaucoma

Summary It is widely accepted that a disturbed blood supply of the optic disc may cause (in addition with an increased intraocular pressure) optic nerve fibre damage. Therefore we measured ocular perfusion pressures in 79 healthy subjects. In 18 patients with low tension glaucoma, in 27 patients suffering from ocular hypertension and in 49 glaucoma (OAG) patients. For measuring perfusion pressures we used the technique of oculooscillo-dynamography (OODG) as described by Ulrich. Additionally we measured intraocular pressure and systemic blood pressure. In OODG the IOP is simultaneously increased in both eyes by application of a suction cup. After increase of the IOP the negative pressure in the suction cup is slowly and linearly decreased. During this decrease the pulse-depending oscillations of each eye are recorded on a strip-chart-recorder. By means of this method retinal and ciliary perfusion pressures can be separated. As a result we could show that mean arterial blood pressure and systolic retinal perfusion pressure were comparable and not statistically significantly different between the groups examined. The systolic ocular perfusion pressures in patients with low tension glaucoma showed a highly statistically significant reduction compared with the other groups. Between healthy subjects, OAG-patients and patients suffering from ocular hypertension there was no difference in systolic ciliary perfusion pressure detectable.