A comparative morphometric study of the myocardium during the postnatal development in normotensive and spontaneously hypertensive rats.

BACKGROUND: Differences in the size of cardiac muscle cells observed in normal and hypertrophic hearts have been assessed through different methodologies. Spontaneously hypertensive rats are often used as an experimental model of essential hypertension in humans, which allows researchers to study the relation between hypertension and cardiac hypertrophy. It has been shown that ventricular hypertrophy in mammals progresses and ventricular failure develops in the end stage of hypertrophy. The aim of the present study was to analyse a number of morphometric markers and compare them between male normotensive Wistar rats (WR) and male spontaneously hypertensive rats (SHR). MATERIALS AND METHODS: The total number of male WR was 15, distributed in five age groups, each containing three animals: 2-week-old; 1-month-old; 3-month--old; 6-month-old; 12-month-old. The male SHR were distributed in two age groups, each containing three animals: 1-month-old (young) and 6-month-old (adult). RESULTS: As aging progressed, both in male normotensive WR and in male SHR we noted a statistically significant increase in the morphometric parameters thickness of the free wall and the cross-sectional area of the cardiomyocytes and their nuclei and a decrease in the cardiomyocytic density in both ventricles. These changes were more pronounced and occurred at an earlier age in SHR. CONCLUSIONS: The present study analyses in detail the alterations in the myocardium of the left and right ventricle, initiated by age-related hypertrophy, as well as hy-pertrophy induced by arterial hypertension. (Folia Morphol 2018; 77, 2: 253-265).

Estradiol inhibits astrocytic GFAP expression in an animal model of neuroinflammation.

There are many animal models for studying different aspects of neurodegeneration. Lipopolysaccharide (LPS) injected in rats intracerebroventricularly induces neuroinflammation quite similar to the inflammatory component of chronic neurodegenerative conditions such as Alzheimer's disease. We used this model to examine the effect of estradiol on neuroinflammation. LPS or pyrogen-free saline were injected intracerebroventricularly (i.c.v.) into the lateral ventricle of male Wistar rats and estradiol was administered (200 micrograms/kg s.c.) 48 h before or 24 h after LPS injection. LPS-induced body weight loss was partially postponed by the treatment, especially in the rats pretreated with estradiol. When analyzing GFAP glial cell morphology in the CA3c area of the hippocampus and corpus callosum, as well as the number of astroglial cells in CA3c and CA1, GFAP expression was found to be reduced. This was true especially in the animals pretreated with estradiol and to a much lesser extent in the posttreated ones. The data support the possible existence of a neuroimmunomodulatory effect of estradiol administration in neurodegenerative conditions, which influences the inflammatory component.

A post-ischaemic single administration of galanthamine, a cholinesterase inhibitor, improves learning ability in rats.

Transient forebrain ischaemia is widely observed in clinical practice. We have examined the effect of a single administration of the cholinesterase inhibitor galanthamine (2mg kg(-1) i.p.) 25 min after reperfusion in male Sprague-Dawley rats (180 +/- 20 g) after a 20-min common carotid artery occlusion. Twenty-four-hours post-ischaemia there was no difference in motor co-ordination or muscle tonus of the rats treated with or without galanthamine as assessed by the rota-rod test. Learning ability was examined using the shuttle-box test, evaluating the latency time and the number of errors for six days in succession. The performance of the ischaemic saline-injected rats was significantly impaired on days 4, 5, 6 (latency time) compared with the non-ischaemic rats and with the ischaemic animals administered galanthamine (P < 0.05). Similar results were obtained when counting the number of errors (failure to cross the cage during conditioned or unconditioned stimulus). The monitoring of body temperature during the first 12-h post-ischaemia did not show any significant difference between the groups. The data showed a beneficial effect of galanthamine on the recovery of learning ability when administered once only post-ischaemia. This suggests a direct effect on the early pathologic mechanisms of CNS damage. Cholinesterase inhibitors may prove useful in the early clinical treatment of ischaemic conditions.

Effect of the acetylcholinesterase inhibitor galanthamine on learning and memory in prolonged alcohol intake rat model of acetylcholine deficit.

Different cholinomimetics are used in conditions of CNS acetylcholine (ACh) deficit. In this study, we examined the effect of the acetylcholinesterase inhibitor galanthamine in a prolonged alcohol intake model of ACh deficit in male Wistar rats. After 16 weeks of alcohol intake and a 2-week pause, rats administered galanthamine (2.5 mg/kg/day i.p.) showed an improved speed of learning and short-term memory in the shuttle box test as compared to the saline-injected alcoholic group (p < 0.05). Four weeks later, significant improvement in the passive avoidance memory of alcoholic galanthamine-treated rats was noted in the eight-arm radial maze (14 day test duration) as compared to the saline-injected alcoholic group (p < 0.05). During the first week in the shuttle box test, the nonalcoholic galanthamine-treated animals exhibited significantly impaired performance as compared to the untreated nonalcoholic control, while four weeks later, in the eight-arm radial maze, both groups did not differ. Our results show that galanthamine improves the speed of learning, short-term memory and spatial orientation of rats in conditions of prolonged alcohol intake.

Effects of dexamethasone in rat neonatal model of axotomy-induced motoneuronal cell death.

In this study the effect of dexamethasone on the motoneuronal cell death and the nuclear and somatic morphology changes occurring after peripheral nerve transection in the neonatal rats has been determined. The study was performed on 3 day old Wistar rats. Animals were divided into 3 groups--control, axotomised, and axotomised and dexamethasone-treated. The nerve transection was performed bilaterally. A dose of 0.5 mg/kg/24h dexamethasone, administered i.p., was used. On day 7 after the operation the animals were sacrificed and the motoneurons in segments L4 and L5 in the spinal cord were counted and their morphology was analysed. 25. 88% cell loss was found in the axotomised group (p<0.001 vs. control) versus 43.33% cell loss in the dexamethasone-treated and axotomised animals (p<0.01 vs. control). Dexamethasone significantly decreased the number of the surviving motoneurons (p<0.05 vs. axotomised). The axotomised group showed enlargement of the somatic area and the maximal and minimal diameters of the cell while the dexamethasone-treated and axotomised group showed soma shrinkage and decrease in the minimal cell diameter. Our results propose a possible hazard towards the application of dexamethasone in the treatment of new-borns with concomitant nerve injuries.

Light chain usage in anti-double-stranded DNA B cell subsets: role in cell fate determination.

Two major mechanisms for the regulation of autoreactive B cells that arise in the bone marrow are functional silencing (anergy) and deletion. Studies to date suggest that low avidity interactions between B cells and autoantigen lead to B cell silencing, whereas high avidity interactions lead to deletion. Anti-double stranded (ds) DNA antibodies represent a pathogenic autospecificity in Systemic Lupus Erythematosus (SLE). An understanding of their regulation is critical to an understanding of SLE. We now demonstrate in a transgenic model in which mice express the heavy chain of a potentially pathogenic anti-DNA antibody that antibody affinity for dsDNA does not alone determine the fate of anti-dsDNA B cells. B cells making antibodies with similar affinities for dsDNA are regulated differently, depending on light chain usage. A major implication of this observation is that dsDNA may not be the self antigen responsible for cell fate determinations of anti-dsDNA B cells. Light chain usage may determine antigenic cross-reactivity, and cross-reactive antigens may regulate B cells that also bind dsDNA.

Studies on the structure, regulation, and pathogenic potential of anti-dsDNA antibodies.

Studies of anti-double-stranded (anti-ds)DNA antibodies have provided insights into how and why these antibodies arise in systemic lupus erythematosus. In this review we discuss the experimental approaches that have been used by our laboratory to study these autoantibodies. Structure/function analyses including site-directed mutagenesis have helped characterize the molecular genetics of anti-dsDNA antibodies, and more recently peptide libraries have been used to define molecular motifs that these antibodies bind. Most of the pathogenic anti-dsDNA antibodies observed in lupus are somatically mutated. We demonstrated in vitro and in vivo that anti-bacterial antibodies can mutate to acquire specificity for dsDNA. Furthermore, using a fusion partner constitutively expressing bcl-2, NSO(bcl-2), we have shown the existence of anergic or preapoptotic B cells making antibodies that cross-react with both bacterial antigen and dsDNA. Whether defects in the regulation of these antibodies might contribute to serum expression of anti-dsDNA antibodies in some individuals remains unknown. A major emphasis of this review is the regulation of anti-dsDNA antibodies in a transgenic mouse model harboring the gene for the heavy chain of a pathogenic anti-dsDNA antibody. Nonautoimmune transgenic mice effectively regulate autoreactive B cells by anergy and deletion, while their autoimmune counterparts do not. The vast majority of anergic B cells expressing high-affinity transgenic anti-dsDNA antibody fail to display allelic exclusion of the heavy chain. We postulate that this may be one mechanism that allows them to escape deletion. Comparative studies on light chain usage in both the autoimmune and the nonautoimmune transgenic mouse strains have demonstrated that within the autoreactive B-cell population, there are subsets that are differentially regulated. Ultimately transgenic animals making pathogenic autoantibodies may provide us with a system for testing novel therapies for autoimmune disease.

Lack of allelic exclusion permits autoreactive B cells to escape deletion.

The R4A-gamma 2b transgenic mouse harbors the gene for the gamma 2b heavy chain of an anti-dsDNA Ab. Approximately 80% of B cells expressing the transgene display allelic exclusion. Although the transgenic mice have little to no detectable serum anti-DNA activity, splenic B cells can be stimulated in vitro with LPS to secrete anti-DNA Ab. Hybridomas derived from LPS-stimulated splenic B cells were analyzed for expression of the transgene and for DNA binding. All nine transgene-encoded anti-DNA-producing lines were found to express an endogenous IgM heavy chain. Of 19 randomly selected lines producing a transgene-encoded non-DNA binding Ab, none expressed a second heavy chain. The tight correlation between lack of allelic exclusion and anti-dsDNA specificity provides strong support for the hypothesis that a major function of allelic exclusion is to prevent the maintenance of a pool of potentially activatable autoreactive cells.

The anti-DNA-associated idiotype 8.12 is encoded by the V lambda II gene family and maps to the vicinity of L chain CDR1.

The 8.12 idiotype is an anti-DNA-associated Id present on lambda L chains that are expressed at high titers in 50% of patients with systemic lupus erythematosus. Since this Id can be present on as much as a third of a patient's anti-DNA antibodies and is found in renal glomeruli, 8.12 is thought to be a marker for a subset of pathogenic anti-DNA auto-antibodies. A molecular analysis of the 8.12 positive antibodies was designed to explore the genetic basis of this Id. Monoclonal human B cell lines were generated by transformation with EBV and lambda L chain-secreting lines were analyzed for Id expression and V region gene usage. In this panel of Ig lambda cell lines, the 8.12 idiotype is encoded exclusively by members of the V lambda II gene family. The sequences of several 8.12+ and 8.12- V lambda II genes are reported here and are used to map the 8.12 Id to the vicinity of CDR1, as well as to further characterize the large and polymorphic V lambda II gene family.

[Microclimate parameter study in a building for the cage battery raising of small pigs]. / Izsledvane na niakoi parametri na mikroklimata v sgrada za kletuchno-bateriino otglezhdane na malki praseta.

An experiment was carried out in a reconstructed building for cell-battery rearing of young pigs, heated by two petroleum calorifers. Six to eight pigs were placed in a cell in 4 row-cell batteries at two storeys. Ventilation was realized by sucking ventilators, situated along the ridge of the building. Temperature and relative air humidity were measured at 7, 14 and 21 o'clock. The gas composition and the air movement rate were periodically investigated. It was established that at temperatures in the region below 0 degrees C regime for that category of pigs cannot be maintained with the accepted heating constructions. The gas composition standards established and the rate of air movement are within the range of hygienic standards. At the level of the second cell and crosswise to the building slighter temperature differences were observed and the temperature was higher. The conclusion is made that the chosen ventilation scheme and equipment for heating do not ensure optimal and uniform temperature-moisture regime in the premises.