Relationship between Plasma Albumin Concentration and Plasma Volume in 5 Inbred Rat Strains.

Using the Evans Blue procedure, we previously found strain-related differences in plasma volumes in 5 inbred rat strains. Because albumin binds strongly with Evans blue, this protein is important in the Evans blue method of plasma volume determination. Therefore, we speculated that interstrain differences in plasma albumin concentration (PAC) could distort calculated plasma volumes. To address this concern, we used ELISA techniques to measure PAC in these inbred rat strains. In study A, the blood volume was measured by using Evans blue dye, and albumin was measured at the start of hemorrhage. In study B, blood volume was not measured, and albumin was measured twice, near the start and end of hemorrhage (approximately 14 min apart). Neither study revealed any interstrain differences in PAC, which decreased after hemorrhage in all 5 strains. No correlation was found between PAC and plasma volume, survival time, blood lactate, or blood base excess. Percentage changes in PAC during hemorrhage were greater in salt-sensitive compared with Lewis rats. Moreover, these percentage changes were associated with survival time in Fawn hooded hypertensive rats. Our data show that the plasma volumes we measured previously were not misrepresented due to variations in PAC.

Arterial blood gases, electrolytes, and metabolic indices associated with hemorrhagic shock: inter- and intrainbred rat strain variation.

We have previously shown interstrain variation (indicating a genetic basis), and intrastrain variation in survival time after hemorrhage (STaH) among inbred rat strains. To assist in understanding physiological mechanisms associated with STaH, we analyzed various arterial blood measures (ABM; pH, Paco2, oxygen content, sodium, potassium, glucose, bicarbonate, base excess, total CO2, and ionized calcium) in inbred rats. Rats from five inbred strains (n = 8-10/strain) were catheterized and, ≈ 24 h later, subjected to a conscious, controlled, 47% hemorrhage. ABM were measured at the start (initial) and end (final) of hemorrhage. Inter- and intrainbred strain variations of ABM were quantified and compared, and correlations of ABM with STaH were determined. All final ABM values and some initial ABM values were different among strains. Most ABM changed (Δ) during hemorrhage, and these changes differed among strains (P <0.03). Some strain-dependent correlations (r ≥ 0.7; P ≤ 0.05) existed between ΔABM and STaH (e.g., BN/Mcwi, ΔK(+), r = -0.84). Dark Agouti rats (longest STaH) had the smallest ΔPaco2, ΔHCO3(-), and Δbase excess, and the highest final glucose. High coefficients of variation (CVs, >10%), strain-specific CVs, and low intraclass correlation coefficients (rI < 0.5) defined the large intrastrain ABM variation that exceeded interstrain variation for most ABM. These results suggest that some ABM (K(+), Paco2, glucose, oxygen content) could predict subsequent STaH in an inbred rat strain-dependent manner. We speculate that whereas genetic differences may be responsible for interstrain variation, individual-specific epigenetic processes (e.g., DNA methylation) may be partly responsible for both inter- and intrastrain ABM variation.

Comparison of the nicotinamide catabolism among rat strains.

We discovered markedly differing catabolism of nicotinamide among rat strains. We compared the catabolism of nicotinamide and also that of the other tryptophan-nicotinamide and water-soluble vitamins among the four strains, Wistar, Sprague-Dawley (SD), August-Copenhagen Irish (ACI) and Fischer 344. The major urinary catabolite of nicotinamide was N(1)-methyl-4-pyridone-3-carboxamide in Wistar, SD and ACI, and N(1)-methylnicotinamide in Fischer rats. This phenomenon was attributed to the enzyme activity involved in the reaction of N(1)-methylnicotinamide to N(1)-methyl-4-pyridone-3-carboxamide being much lower in Fischer than in the other three strains. With the water-soluble vitamins, this specific phenomenon was only observed in the catabolism of vitamin B(6); the urinary catabolite, 4-pyridoxic acid, was much lower too. It was found for the first time that the activities of oxidase were lower in Fischer than in the other strains. This study showed that Wistar, SD, ACI strains had similar water-soluble vitamin metabolism including nicotinamide catabolism.

A prothrombotic phenotype in the Copenhagen rat strain.

INTRODUCTION: Experimental and epidemiologic studies have identified several potential genetic components for increased thrombotic risk. Studies of thrombosis often use rat models without considering the effect of strain differences on thrombotic propensity. MATERIALS AND METHODS: A comparison of in vivo thrombotic occlusion after small-vessel anastomosis was made between age/weight-matched male Copenhagen and Lewis rats. RESULTS: One-day thrombotic occlusion rates were significantly higher in Copenhagen arteries (67%) and veins (100%) compared to Lewis arteries (8%) and veins (50%), respectively. Single-bolus intravenous heparin (150 units/kg body weight) had a slight effect on reducing occlusion rates in Copenhagen rats (50% and 67% for arteries and veins, respectively), while occlusion was totally prevented by heparin in both vessel types of Lewis rats (0% occlusion). In vitro assays for platelet aggregation and coagulation revealed no apparent differences between these two rats strains, although AT-III levels were slightly higher in Copenhagen rats, contrary to the prothrombotic state. CONCLUSIONS: These findings indicate a profound prothrombotic tendency in the Copenhagen rat strain and support a broader investigation of the genetic basis of this thrombotic potential.

Thermoregulatory responses of the inbred heat-tolerant FOK rat to cold.

The responses of inbred heat-tolerant FOK rats to cold were compared with those of Wistar King A/H (WKAH) and Std:Wistar (WSTR) strains. The fall of colonic temperature during cold exposure was unexpectedly smaller in FOK than in other groups, but the onset of shivering was delayed in FOK. Norepinephrine (NE)-induced in vivo oxygen consumption and the mitochondrial uncoupling protein 1 level of brown adipose tissue (BAT) were not different among the groups, but the cold-induced increases in in vivo oxygen consumption as well as plasma glycerol and free fatty acids were higher in FOK than in other groups. In vitro NE-induced oxygen consumption of BAT was less in FOK than WSTR, but not WKAH. The magnitude of the NE-induced increase in blood flow through BAT was higher in FOK than in other groups. These results suggest that FOK paradoxically have a high capacity for nonshivering thermogenesis in spite of their high capacity for heat tolerance, probably due to an increased lipid utilization and improved circulation of BAT.

Angiotensin I-converting enzyme genotype influences arterial response to injury in normotensive rats.

Two normotensive strains of rat, the Lou and Brown Norway (BN) strains, have contrasting levels of plasma angiotensin-converting enzyme (ACE). To investigate the degree of genetic determination of ACE expression, a polymorphic marker of the ACE gene was analyzed in inbred rats of the two strains. The two inbred strains were shown to bear different alleles for a polymorphic marker at the ACE gene. The segregation of the alleles of this marker and the plasma ACE levels were studied in a group of F2 rats issued from a cross between Lou and BN rats. The degree of genetic determination of plasma ACE activity was estimated to be 94% in the F2 cohort. The ACE locus accounts for 74% of total plasma ACE variance. ACE activity and mRNA expression in lungs were also genetically determined. The difference observed in ACE mRNA accumulation in the lungs between the two strains was due to a difference in the transcriptional rate of the ACE gene, as shown in nuclear run-on experiments. No differences were observed in arterial blood pressure of homozygous F2 progeny. In these animals, ACE genotype did not interfere with the pressor or the depressor responses to ACE-dependent vasoactive peptides. There was a significant effect of strain on constitutive or inducible membrane or soluble ACE activity in primary cultures of vascular cells. Neointima formation in the carotid artery 14 days after balloon injury was also influenced by the genotype in F2 homozygous progeny, whereas the medial area was not. These results demonstrate that there is a close relationship between the genetically determined ACE expression and the inducibility of the ACE gene. The degree of genetic determination of ACE expression in inbred rat strains offers a unique opportunity to study the interaction between genetic and environmental determinants of ACE expression and its involvement in response to experimental cardiovascular and renal injury.

Extrahepatic C6 is as effective as hepatic C6 in the generation of renal C5b-9 complexes.

In order to study the contribution of extrahepatic C6 to anti-Thy1.1 nephritis, C6 deficient PVG/c- livers were grafted in C6 sufficient PVG/c+ rats (Tx-L). Infusion of anti-Thy1.1 antibodies in Tx-L and PVG/c+ rats resulted in generation of C5b-9 complexes and subsequent glomerular injury, while infusion of anti-Thy1.1 antibodies in PVG/c- rats revealed no detectable C6 deposition. Because C6 mRNA was expressed in both liver and kidney tissue of PVG/c+ rats, we assessed whether production of C6 in the kidney alone was sufficient for glomerular injury. One kidney of a PVG/c- rat was replaced with a PVG/c+ kidney (Tx + K) followed by administration of anti-Thy1.1 antibodies. C6 deposits were detectable neither in PVG/c+ kidneys nor in PVG/c- kidneys of Tx + K rats, indicating that C6 production in PVG/c+ kidneys alone is not sufficient to contribute to renal injury. That C6 production had occurred was suggested by the presence of equal amounts C6 mRNA in control PVG/c+ kidneys and in grafted PVG/c+ kidneys of Tx + K rats. C6 mRNA expression in kidney tissue of PVG/c+ rats is presumably derived from peritubular sites. In conclusion, we have demonstrated that extrahepatic, but not renal synthesis of, C6 is sufficient to contribute to glomerular injury during anti-Thy1.1 nephritis.

Vitamin E ameliorates the renal injury of Dahl salt-sensitive rats.

Recently, hyperlipidemia as well as hypertension has been observed in Dahl salt-sensitive (S) rats. In this study, to investigate whether the lipid abnormality is involved in the renal injury of Dahl S rats, we examined the effect of vitamin E on glomerular sclerosis, as vitamin E is an inhibitor of lipid oxidation. Dahl S rats were given a high salt diet (8% NaCl) containing either normal vitamin E (2 mg/100 g) or high vitamin E (50 mg/100 g) for 4 weeks. Dahl salt-resistant (R) rats were given a high salt and normal vitamin E diet. The blood pressure in the Dahl rats increased and was not suppressed by the vitamin E supplement. Serum cholesterol and triglycerides in Dahl S rats were higher than in Dahl R rats at both 0 and 4 weeks. Vitamin E lowered the serum cholesterol level in Dahl S rats at 4 weeks (126 +/- 5 v 150 +/- 12 mg/dL, P < .01). Urinary protein excretion and serum creatinine increased in Dahl S rats, and vitamin E inhibited the increases significantly (urinary protein, 70.7 +/- 0.9 v 178.0 +/- 8.8 mg/day, P < .01; serum creatinine, 0.45 +/- 0.02 v 0.63 +/- 0.05 mg/dL, P < .01). Serum lipid peroxide (LPO) was higher in Dahl S rats than in Dahl R rats, and vitamin E lowered LPO in Dahl S rats (2.10 +/- 0.03 v 2.70 +/- 0.04 nmol/mL, P < .01). In the histologic study, sclerosing score (SS) of glomeruli, which represents the degree of glomerulosclerosis semiquantitatively, was higher in Dahl S rats than in Dahl R rats. Vitamin E lowered SS (114 +/- 3 v 157 +/- 6, P < .01) and ameliorated arterial injuries such as medial thickness with partial necrosis and severe fibrinoid proliferation with inflammatory cell infiltration. In all rats, SS was strongly correlated with urinary protein (r = 0.93, P < .01), serum cholesterol (r = 0.86, P < .01), and serum LPO (r = 0.89, P < .01). These results suggest that the renal injury in Dahl S rats is caused not only by hypertension but also by hyperlipidemia. Therefore, vitamin E might ameliorate the renal damage by inhibiting the oxidation of lipids.

Strain variation in vitamin A (retinol) status of streptozotocin-induced diabetic rats.

Decreased plasma concentrations of vitamin A (retinol) and retinol-binding protein have been previously identified in human subjects with type I diabetes mellitus. The present study was undertaken to investigate the effects of streptozotocin-induced diabetes in rats of three different strains including Wistar Furth, Sprague Dawley and Wistar, on plasma and liver concentrations of vitamin A. The diabetic animals gained less weight than nondiabetic controls even though they ate 50% more food. The hepatic vitamin A concentration was increased at three weeks after the onset of diabetes in all three strains of rats but the magnitude of increase was greater in Wistar than either Wistar Furth or Sprague Dawley rats. This increased storage of vitamin A in diabetic animals most likely is due to increased food intake. The plasma concentrations of vitamin A, on the other hand, remained unaffected in Wistar Furth and decreased moderately (P < 0.02) in Sprague Dawley but severely (P < 0.0001) in Wistar rats. The fact that the plasma vitamin A levels in diabetic Wistar and Sprague Dawley rats were markedly reduced despite their increased hepatic store suggest an impairment in the transport of vitamin A from the liver. The circulatory levels of vitamin A in Wistar rats are more sensitive to the diabetic state, which is in agreement with those observations seen in diabetic patients. Because of this similarity, it is reasonable to suggest that Wistar should be the choice of rat strain for future experimental studies involving vitamin A and diabetes relationships.

Plasma adenosine concentrations are elevated in Dahl salt-sensitive rats.

We measured plasma levels of adenosine in Dahl salt-sensitive rats (DS) and Dahl salt-resistant rats (DR) to examine the potential role of adenosine in cardiovascular regulation in this type of hypertension. Plasma adenosine concentrations were significantly higher in DS than in DR. The NaCl content in the diet did not affect plasma adenosine concentration in either DS or DR. Significant positive correlation was found between adenosine concentrations and systolic blood pressure when the data for DS and DR were analyzed together. These results suggest that adenosine may play an important role in the pathophysiology of hypertension in DS.

Disposicao cinetica da ciclosporina apos administracao de dose unica: estudo experimental em ratos / Kinetic disposition of the ciclosporin after single dose subcutaneous administration: experimental study in rats

A dose adequada de ciclosporina e aquela que apresenta satisfatorio efeito imunossupressor associado a baixa toxicidade. Na literatura, as doses utilizadas variam de 2,5 mg/kg a 25 mg/kg em trabalhos experimentais de alotransplantes de tecidos e orgaos. Preferencialmente sao administradas pelas vias subcutanea ou intra-muscular, com intervalos de 24,48 horas e ate sete dias. Neste estudo utilizou-se para a administracao de dose unica de ciclosporina A (CSA), a via subcutanea em 21 ratos que foram divididos em tres grupos com sete animais cada, recebendo 2,5 mg/kg (grupo 1), 5,0 mg/kg (grupo 2) e 10,0 mg/kg (grupo 3). Colheram-se amostras de sangue nos tempos 1, 4, 8, 12, 24, 48 e 72 apos a administracao da droga. Efetuou-se ciclosporinemia pelo metodo de radioimunoensaio, Kit SandimmunR monoclonal especifico...

Chimeric drift in blood cell populations of chimeric rats constructed between congenic strains.

Chimeric drift is the shift in the proportion over time of the two cell lineages which comprise a chimera (genetic mosaic). Chimeric drift in blood cell populations is determined by both the probability of proliferation from stem cell pools of one or the other of the cell lineages which constitute the chimera and the effects of life span in circulating blood cells. Previous evidence suggests that while chimeric drift occurs in chimeras between genetically disparate strains, it does not occur when the strains used are closely related. No information is available from chimeras between congenic strains. In the present study, chimeric rats were produced between strains with distinguishable class I major histocompatibility complex haplotypes, PVG-RT1a and PVG (which express the haplotype RT1c). PVG-RT1a-specific monoclonal antibodies were used to establish the mosaic patterns in the cell populations of peripheral blood by fluorescein-activated cell sorting. Mosaic cell lineage of red blood cells, white blood cells, lymphocytes, monocytes and neutrophil populations were analyzed weekly over a period of 6 weeks. The ratio of cells of the PVG-RT1a lineage to cells of the PVG lineage shifted either in favor of PVG-RT1a or PVG in cellular components of peripheral blood. The percentage of PVG-RT1a cells in peripheral blood of chimeras changes by as much as 54, 28, 21, 19 and 23% in red blood cell, white blood cell, lymphocyte, monocyte and neutrophil populations, respectively. The shifts in the percentage of PVG-RT1a cells appears to occur in a cyclic fashion.(ABSTRACT TRUNCATED AT 250 WORDS)

[Hematological and serum biochemical changes with age in term fetuses, offspring and dams in normal Sprague-Dawley rats].

Hematological and serum biochemical values of dams and offspring of Sprague-Dawley rats were measured during late gestation, lactation and postweaning. In dams, slightly low erythrocytic parameters, high platelets and high frequency of neutrophils were seen after parturition although WBC showed no marked changes. On Day 20 of gestation, glucose and triglycerides were extremely high and TP and albumin were low. These changes may be attributed to pregnancy or parturition. In fetuses on Day 20 of gestation and offspring immediately after birth, erythrocytes showed anisocytosis, polychromasia, basophilic stipplings and Howell-Jolly bodies and erythroblasts were found. RBC was low. MCV and MCH were extremely high, compared to adult erythrocyte levels. Hemoglobin and hematocrit slightly decreased before weaning. RBC, hemoglobin and hematocrit increased with age and reached adult levels by Day 56. MCV and MCH values decreased, towards adult levels, until weaning. Platelets rapidly increased and reached adult levels before weaning. WBC increased after birth having higher counts in males than in females on Day 35 and thereafter. Glucose, TP and albumin increased with age and reached adult levels by Day 28. ALP was high in fetuses and changed with age having two peaks similar to those reported in man. Cholesterols gradually increased after birth and had a peak on Day 14. Urea nitrogen, inorganic phosphorus and calcium were slightly high in fetuses and preweaned offspring. Potassium was high in fetuses but no age-related trends were seen in offspring.

Pyruvate transport across the plasma membrane of the bloodstream form of Trypanosoma brucei is mediated by a facilitated diffusion carrier.

The characteristics of pyruvate transport across the plasma membrane in the bloodstream form of Trypanosoma brucei were studied using [14C]pyruvate in combination with the silicone-oil centrifugation technique. We present evidence for the existence of a facilitated diffusion carrier in the plasma membrane of T. brucei which specifically mediates the translocation of pyruvate. The uptake of pyruvate followed saturation kinetics (Km 1.96 +/- 0.28 mM; Cmax 36.61 +/- 1.15 nmol pyruvate/30 sec.mg protein), after correction of the data for a nonsaturable diffusion component. The uptake of pyruvate was competitively inhibited by a number of (oxo)monocarboxylic acids, including pyruvate analogs and metabolically related substances, but not by L-lactate. The transport exhibited the phenomenon of transacceleration, indicative for the involvement of a facilitated diffusion carrier. The carrier is highly specific for pyruvate and differs from other known monocarboxylate carriers present in the mitochondrial and/or plasma membrane of other eukaryotic cells in that it does not transport L-lactate.

Comparison of hematologic parameters in normal and streptozotocin-induced diabetic rats.

Hematologic values are compared for normal and streptozotocin-induced diabetic rats after 6 weeks of induced diabetes. Most hematologic parameters were the same in the two groups except for blood glucose, glycated hemoglobin, and 2,3 diphosphoglycerate, all of which were elevated in the streptozotocin group. However the P50 (the PO2 at which the oxygen-carrying capacity of blood is 50% of maximal) remained normal. We hypothesize that a left shift in the oxyhemoglobin dissociation curve caused by the glycation of a small percentage of the hemoglobin is compensated by elevation in the 2,3-diphosphoglycerate which returns the P50 to normal values. This compensatory mechanism also occurs in some stages of human diabetes.

Changes in osmotic resistance of erythrocytes of cattle, pigs, rats and rabbits during variation in temperature and pH.

The osmotic resistance of cattle, pig, rat and rabbit erythrocytes was determined at different temperatures and pH. At constant pH of 7.7, the resistance of erythrocytes of all these species increased as the temperature increased. However, at constant temperature of 29 degrees C, the resistance of the erythrocytes decreased in acidic solution and increased in alkaline solution.

[Number and maturation distribution of the reticulocytes from six animal species]. / Zahl und Reifegradverteilung der Retikulozyten von sechs Tierarten.

With the fully automated reticulocyte analyzer Sysmex R-1000 (Fa. Toa Medical Electronics Co., LTD) the reticulocyte count and the reticulocyte ratio of rabbit, rat, mouse, mastomys, dog and swine was determined. Simultaneously, the reticulocytes were subdivided into 10 maturation classes according to their fluorescence intensity. The maturation distribution can be characterized by a distribution quotient. Both the maturation distribution as well as the distribution quotient are considered to be useful as good indicators of erythropoietic activity. The investigation establishes, that the reticulocyte count and the maturation distribution of these six species can be determined quickly and easily. The distinct differences between the species reflect the maturation locus of the reticulocytes, which varies among the species (medullary or humoral type of maturation).

[Evaluation of blood chemistry parameters of rats treated with 2-acetylaminofluorene and N-acetylcysteine]. / Valutazione di parametri ematochimici in ratti trattati con 2-acetilamminofluorene e N-acetilcisteina.

Male wistar rats were treated with a diet supplemented with 0.05% 2-acetylaminofluorene (2AAF) and/or 0.2% N-acetylcysteine (NAC) according to the protocol of Teebor and Becker. Eleven haematochemical parameters were evaluated at the third week of the first two cycles. The results showed a slight yet significant decrease in total proteins and triglycerides, and an increase in total bilirubin, gamma-glutamyltranspeptidase and alkaline phosphatase, as compared to untreated controls. Co-treatment with NAC slightly attenuated the alterations induced by 2AAF. On the whole, these results demonstrate that 2AAF is poorly necrotic to hepatocytes, and hence its known ability to damage the liver appears to mainly depend on nuclear effects rather than on cytoplasmic changes.