Effect of different concentrations of stem juice of Oxalis tuberosa Mol. and juice of the fruit of Gaultheria glomerata (Cav.) Sleumer on the antioxidant activity of the heat-treated functional beverage.

Functional beverages with added health benefits are popular among peoples and athletes because they help them recover faster from intense workouts and perform better overall. This research set out to determine how well heat-treated stem juice from Oxalis tuberosa Mol. "oca" and fruit juice from Gaultheria glomerata (Cav.) Sleumer "laqa-laqa" performed as an antioxidant in a functional drink. The "oca" stems and the "laqa-laqa" fruit were collected to obtain the juice. For this study, 30 semi-trained panellists used sensory evaluation to rate four treatments (Bo, B1, B2, and B3) with varying quantities of "oca" and "laqa-laqa" juice. The results concluded that the treatment B2, which included 300 ml of "oca" stem juice, 800 ml of "laqa-laqa" juice, 1000 ml of treated water, and 220 g of refined sugar, was given the highest score after a physicochemical evaluation of its colour, smell, taste, and overall appearance. Similarly, the results showed that the protein content increased by 1.38%, the fat content by 1.08%, the moisture percentage by 99.5%, the ash content by 1.82%, and the carbohydrate content by 6.22% after B2 treatment. Similarly, results revealed significant enhancement in antioxidant profiling such as total polyphenols: 1825 mg of gallic acid/100 g and antioxidant Activity: 89.56% µmol of trolox /100 g. In conclusion, due to its high energy content and antioxidant activity, it may be a viable nutritional option for athletes who engage in rigorous, frequent physical exertion.

The pigment of melanosis coli: a lectin histochemical study.

BACKGROUND: The compositional nature of the pigment of melanosis coli is essentially unknown. Previous histochemical studies suggested that this pigment has certain similarities with lipofuscin (i.e., age-dependent pigment) and ceroids (i.e., pathologically derived pigments) and that it may contain, therefore, polymerized glycolipids and glycoproteins. However, the saccharide composition of this pigment was never explored by lectin histochemical procedures, which was the main object of this study. METHODS: Colonoscopic biopsy specimens from eight patients with melanosis coli and from three normal control subjects were studied by fluorescent microscopy and by standard and lectin histochemistry. The number of apoptoses in the lining colonic epithelium was also evaluated histologically. RESULTS: Apoptotic bodies were significantly more numerous in patients with melanosis coil than in control subjects. The pigment that accumulates in macrophages of the lamina propia showed autofluorescence, sudanophilia, acid-fastness, and positiveness to PAS and Schmorl's reactions, all of which are common to lipofuscin and ceroids, plus an intense argentaffin reaction abolished by bleaching, indicative of a melanic substance. Lectin histochemistry showed, in decreasing order of frequency, the presence of alpha-D-mannose, sialic acid, beta-D-galactose (lactose), gal-beta-(1-3)acetyl-galactosamine, alpha-D-galactose, and alpha-L-fucose, but no terminal alpha-D-acetyl-galactosaminyl residues. CONCLUSIONS: The significant increase of apoptotic bodies in the lining colonic epithelium indicated that this type of cell death is not due to the natural programmed cell renewal, but to the action of laxatives. Because the autofluorescent pigment of melanosis coli contains melanin (as well as glycoconjugates) and is not dependent on age but on the use of anthranoid laxatives, it should be categorized as a "melanized ceroid." The lectin affinities of this pigment indicated that it contains a substantial number of saccharide residues almost similar to those found in the ceroid pigment of human aortic atheromas. These findings and considerations on the metabolism and pharmacokinetics of anthranoids suggested that the apoptotic epithelial cells, rather than the laxatives, may be the source of the pigment saccharides, whereas the precursors of the melanic substance may be derived from the anthranoids.

Dietary modulation of oxidative stress in alcoholic liver disease in rats.

Numerous studies dealing with prolonged feeding of rats with ethanol liquid regimens high in fat and low in carbohydrate showed that the resulting hepatic pathologic changes, including increased lipid peroxidation, are due to dietary aberrations rather than to ethanol toxicity. The amount and particularly the type of dietary fat significantly modulate the hepatic oxidative stress and morphofunctional reactivities. Although dietary vitamin E modulated oxidative stress or lipid peroxidation, it did not influence the development of hepatic pathologic changes in different animal models of chronic alcoholism. The old observation that lipotropes modulate the hepatic alterations associated with prolonged excessive ingestion of ethanol has been amply confirmed by even those who for years did not accept the importance of lipotropes. Our recent studies in rats indicated that prolonged feeding of large amounts of ethanol and diets with variable amounts of lipotropes, vitamin E and minerals did not significantly modulate a large series of hepatic prooxidants, but decreased several antioxidants (vitamin E, ubiquinols and glutathione peroxidase). Ethanol regimens relatively low in vitamin E increased the hepatic thiobarbituric acid-reactive substances and chemiluminescence and reduced some of the antioxidant factors. However, the hepatic prooxidant factors were unaffected, and no liver damage was detected. These and other findings indicated that the eventual detection of oxidative stress in experimental alcoholic liver disease primarily depends on the type of diet and that oxidative stress may not play a significant pathogenic role in this condition.

In situ rat brain and liver spontaneous chemiluminescence after acute ethanol intake.

The influence of acute ethanol administration on the oxidative stress status of rat brain and liver was assessed by in situ spontaneous organ chemiluminescence (CL). Brain and liver CL was significantly increased after acute ethanol administration to fed rats, a response that is time-dependent and evidenced at doses higher than 1 g/kg. Ethanol-induced CL development is faster in liver compared with brain probably due to the greater ethanol metabolic capacity of the liver, whereas the net enhancement in brain light emission at 3 h after ethanol treatment is higher than that of the liver, which could reflect the greater susceptibility of brain to oxidative stress. The effect of ethanol on brain and liver CL seems to be mediated by acetaldehyde, due to its abolishment by the alcohol dehydrogenase inhibitor 4-methylpyrazole and exacerbation by the aldehyde dehydrogenase inhibitor disulfiram. In brain, these findings were observed in the absence of changes in the activity of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, and glucose-6-phosphate dehydrogenase. However, the content of brain glutathione was significantly decreased by 31%, by ethanol, thus establishing an enhanced oxidative stress in this tissue.

Prooxidant and antioxidant hepatic factors in rats chronically fed an ethanol regimen and treated with an acute dose of lindane.

While acute lindane treatment and chronic ethanol feeding to rats have been associated with hepatic oxidative stress, the possible roles of these stresses in the pathogenesis of hepatic lesions reported in acute lindane intoxication and in those observed in some models of chronic alcoholism have not been established. Our previous studies in rats chronically fed ethanol regimens and then treated with a single intraperitoneal (i.p.) dose of lindane (20 mg/kg) showed that while lindane per se was invariably associated with hepatic oxidative stress, chronic ethanol feeding only produced this stress when the dietary level of vitamin E was relatively low. Chronic ethanol pretreatment did not significantly affect the lindane-associated oxidative stress, and neither chronic ethanol feeding nor acute lindane, single or in combination, produced any histologic and biochemical evidence of liver damage. In the present experiment, the acute dose of lindane was increased to 40 mg/kg, and we have studied a larger number of prooxidant and antioxidant hepatic factors. Male Wistar rats (115.5 +/- 5.4 g) were fed ad lib for 11 weeks a calorically well-balanced and nutritionally adequate basal diet, or the same basal diet plus a 32% ethanol/25% sucrose solution, also ad lib, and were then injected i.p. with a single dose of lindane or with equivalent amounts of corn oil. The results indicated that acute lindane treatment to naive rats increased practically all the prooxidant hepatic factors examined (cytochromes P450 and b5, NADPH cytochrome c reductase, NADPH oxidase), as well as the generation of microsomal superoxide radical and thiobarbituric acid reactive substances of liver homogenates, but did not modify any of the antioxidant hepatic factors studied. Conversely, the chronic administration of ethanol alone did not significantly affect the prooxidant hepatic factors but reduced some of the antioxidants (i.e., the activities of GSH-Px and the contents of alpha-tocopherol and ubiquinols 9 and 10). Although chronic ethanol pretreatment further increased the superoxide generation induced by lindane per se, it did not increase but generally reduced the effects of lindane per se on the other prooxidant factors studied. Furthermore, although acute lindane administration to ethanol-pretreated rats was associated with decreases in GSH and catalase (not affected by ethanol or lindane treatment alone), it did not substantially modify the reducing effects of ethanol feeding per se on GSH-Px, alpha-tocopherol, and ubiquinols. Once again, neither chronic ethanol feeding nor lindane treatment, single or in combination, was associated with any evidence of liver damage.

Lectin histochemistry of lipofuscin and certain ceroid pigments.

Little is known at present about the saccharide components of lipofuscin (age pigment) and ceroid pigments in situ. The purpose of this study was, therefore, to study in detail the lectin reactivities of lipofuscin in neurons and cardiac myocytes of old humans and rats. In addition, those of diverse ceroid pigments found in human aortic atheromas, in the livers of choline-deficient rats, in the uteri of vitamin E-deficient rats and in the crushed epididymal fat pad of rats, are included. Cryostat and deparaffinized sections from all these tissues were either extracted with a solvent mixture of chloroform-methanol-water (10:10:3, v/v) and incubated with 7 different biotinylated lectins or left untreated. Delipidation was done in order to study whether it was possible to discriminate between the saccharide moieties of glycolipids and glycoproteins of lipofuscin and ceroid pigments in situ. Other similarly treated sections were used to study the autofluorescence, sudanophilia, acid-fastness and reactivity to PAS. The frequency and intensity of lectin binding and standard histochemical properties of all the pigments were evaluated semi-quantitatively and blind. The results indicated that mannose was in general the most consistently detected sugar residue in lipofuscin granules of humans and rats, and that this pigment may also contain acetylglucosamine, acetylgalactosamine, sialic acid, galactose and fucose. However, notable differences were found not only in the lipofuscin saccharide components of different cell types of humans and rats, but also in those in the same type of cells in both species. Although mannose was not detected in the hepatic ceroid of choline-deficient rats, this saccharide moiety was almost always present in the other ceroid pigments. Each of the ceroids also contained other types of saccharides although the frequency of the latter varied between different ceroid pigments. While lipofuscin and each of the ceroid pigments showed somewhat different lectin binding patterns, the variability in the frequency of reactivity to lectins suggests that these patterns may not be permanent but transient. In this sense, it appears that lectin histochemistry may not allow these pigments to be differentiated. Furthermore, the extractive procedures used in this study did not enable us to determine whether the saccharides detected in the pigments in situ corresponded to glycolipids or glycoproteins.

Advances in age pigment research.

Although it is presently accepted that lipofuscin (age-pigment) is the end product of the physiological decay of the cells' own constituents, the intimate mechanisms involved in its formation are largely unknown. The advances in the field of lipofuscinogenesis have been relatively slow, mainly due to the persistent confusion between the naturally occurring normal lipofuscin and the pathologically formed ceroid pigments. Therefore, attempts have been made in this presentation to review first the differential features between these pigments and second, to provide a general overview on the physicochemical properties of lipofuscin. The two prevailing theories on lipofuscinogenesis, the peroxidative theory and the proteolytic decline theory, are critically discussed, and future lines of research are suggested for the resolution of present uncertainties on lipofuscinogenesis. Since lipofuscin is properly considered the hallmark of cellular aging, it is expected that the unraveling of the mechanisms involved in lipofuscin formation will provide important clues to the still unknown underlying causes of cellular aging.

Effects of lovastatin and leupeptin on ceroidogenesis of vitamin E-deficient and -supplemented young rats.

Previous studies in young normal rats have shown that intracerebral administration of the proteinase inhibitor, leupeptin, caused a rapid accumulation of lipofuscin-like pigment in lysosomes of brain cells (Ivy et al., 1984a). On the other hand, we have recently found that the administration of lovastatin, an inhibitor of HMG-CoA reductase, reduced the ceroid-like pigment and dolichol contents in the crushed epididymal fat pad of rats (Porta et al., 1988). In order to study now the possible modulating effects of these enzyme inhibitors on ceroidogenesis associated with vitamin E deficiency, two main groups of weanling Wistar female rats were respectively fed ad libitum a vitamin E-deficient basal diet, or the same diet supplemented with 16 mg% of dl-alpha-tocopherol acetate. The vitamin E-deficient and -supplemented rats were further subdivided and received for 8 weeks their diets alone or with 2, 1, or 0.5 g of lovastatin/kg of diet. Other subgroups were treated with constant peritoneal infusion of 0.5 mg/day of leupeptin by means of osmotic minipumps (Alzet 2002) consecutively implanted at days 15, 30, and 45. Lovastatin treatment to vitamin E-deficient rats was associated with dose-dependent toxicity, resulting in 100%, 75%, and 50% mortality at concentrations of 2, 1, and 0.5 g/kg diet, respectively. This mortality was mainly due to extensive hepatic necrosis. Food intake and growth rates were reduced, while the relative weights of liver, kidneys, spleen, heart and brain, as well as the serum levels of GPT and GOT were significantly increased over the values of the untreated vitamin E-deficient control rats. The volumetric densities of ceroid pigment and the dolichol contents in liver and kidneys were not significantly modified. Lovastatin toxicity was partially prevented by vitamin E supplementation. However, in these supplemented rats, lovastatin treatment did not modify the volumetric densities of hepatic and renal ceroid, although the contents of hepatic and renal dolichol were significantly increased. No correlations could be found between levels of hepatic or renal ceroid and total dolichol content in vitamin E-deficient and supplemented rats. Leupeptin treatment to vitamin E-deficient rats only slightly reduced food intake and growth rates, and did not significantly modify the relative organ weights or the serum levels of cholesterol, GOT and GPT. Although in both vitamin E-deficient and -supplemented rats the leupeptin treatment consistently showed a tendency to increase the volumetric densities of hepatic and renal ceroid pigment, the differences with the control untreated rats were not statistically significant.(ABSTRACT TRUNCATED AT 400 WORDS)

Evaluation of the effect of glucan on hepatic, renal and mammary tumorigenesis in rats.

Glucan was evaluated for its ability to modify the hepatic and renal tumorigenesis induced in partially hepatectomized Sprague-Dawley female rats by diethylnitrosamine (DEN) and phenobarbital (PB), and the mammary tumorigenesis induced in intact Sprague-Dawley female rats by N-nitrosomethylurea (NMU). In both models the rats received every two weeks i.v. glucan (10 mg/kg) or equivalent amounts of dextrose. The results indicated that glucan did not significantly modify the incidence of the chemically-induced hepatic, renal and mammary tumors.

Chronic alcoholism enhances hepatocarcinogenicity of diethylnitrosamine in rats fed a marginally methyl-deficient diet.

To determine whether the chronic consumption of ethanol was capable of enhancing the hepatocarcinogenic activity of diethylnitrosamine per se, or through the accentuation of a methyl deficiency, two groups (A and B) of Sprague-Dawley female rats were fed for 10 months either a 20% casein basal diet marginally deficient in methyl, or the same diet supplemented with choline (1 gm per 100 gm) and folic acid (0.54 mg per 100 gm). Both groups were offered a drinking ethanol solution, while two other nonalcohol control groups (C and D) were isocalorically pair-fed to Groups A and B, and received diets in which the alcohol consumed by the corresponding groups was replaced by isocaloric amounts of sucrose. A baseline nonalcohol Group E, isocalorically pair-fed to Group A, received the intact basal diet of Group A and water. One day before the initiation of the experiment, and again 2 months later, all rats from the five groups were injected with a single i.p. dose of diethylnitrosamine (100 mg per kg). The growth attained by all groups was statistically similar. Hepatic triglycerides in Group A were significantly higher than in all the other groups. While in Group A primary hepatocellular carcinomas and renal tumors were encountered at the end of the experiment in 3 of 6 and in 2 of 6 rats, respectively, no malignancies were observed in any of the other groups. These results indicate that chronic ethanol consumption enhances the hepatocarcinogenic and renal tumorigenic activity of diethylnitrosamine, and strongly suggest that this action is mediated through the accentuation of methyl deficiency.

Effects of dietary selenium on hepatic and renal tumorigenesis induced in rats by diethylnitrosamine.

Seven groups of female Sprague-Dawley rats (approximately 200 gm initial body weight) were injected i.p. with a single subcarcinogenic dose of diethylnitrosamine (40 mg per kg body weight) between 8 to 10 hr after partial hepatectomy, and after a recovery period of 3 weeks (herein called induction stage) received 0.05% phenobarbital in the diet for the rest of the experiment (promotion stage). The rats were fed a 20% casein-based diet containing 0.16 ppm of selenium or the same diet supplemented with 4 or 6 ppm of selenium as sodium selenite. The effects of these three dietary regimens were tested when administered 9 to 11 days before and during induction, 1 week before and during promotion or during the entire experiment. Pair-feeding conditions were used to minimize influences due to differences in food intake and growth. Despite similarities in food intakes, the growth rates in groups receiving the 6 ppm-selenium diet during promotion or during the entire experiment were in general significantly lower than in rats fed the 4 ppm-selenium diet or the 0.16 ppm-selenium basal diet. Survival rates were also significantly reduced in rats fed the 4 and 6 ppm-selenium diets during promotion or during the entire experiment. In rats killed at the 19th week for interim assessment of the experiment's progress, the stereologically analyzed numerical and volumetric densities of hepatic premalignant hyperplastic nodules did not differ significantly between groups. All the remaining rats were killed at the 46th week.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of selenium supplementation on hepatocarcinogenesis in rats.

Four groups of weanling male Wistar rats (Groups A-D) received diethylnitrosamine (DEN, 40 ppm) in their drinking water for four weeks; after a recovery period of two weeks, they received (for the rest of the experiment) phenobarbital (PB, 500 ppm) added to a Torula yeast-based diet containing 0.17 ppm of selenium. Dietary selenium (2 ppm), as sodium selenite, was given to Group B one week before and during DEN treatment, to Group C one week before and during PB treatment, and to Group D during the entire experiment. Groups A and E received the unsupplemented diet, whereas Group E was not treated with DEN or PB. Pair-feeding conditions were used to minimize possible influences of differences in food intake and growth. Rats were killed at the 19th and 24th weeks after the experiment began. No significant differences were found in food and fluid intakes or in growth rates among the groups. Livers in Group E were histologically normal, whereas preneoplastic and neoplastic lesions were found in all other groups. In rats killed at the 19th and 24th weeks, the numerical and the volumetric densities of preneoplastic lesions did not differ significantly between all the groups. Similarly, the incidence of hepatocellular carcinomas only detected at 24 weeks was not significantly different between the groups. These results indicated that in this particular model of hepatocarcinogenesis, the dietary supplementation of 2 ppm of selenium did not modify the development of preneoplasia and carcinomas.

Acute adriamycin cardiotoxicity in rats.

To explore the possible occurrence and pathogenic implications of in vivo heart lipoperoxidation in the acute model of ADR-cardiotoxicity, male Wistar rats were injected i.v. with a single dose of ADR (15 mg/kg) and the controls with saline. The rats were killed at 24 and 96 hr after treatment and at the later period the serum levels of creatine kinase of ADR-treated rats were significantly elevated. ADR-treatment did not significantly modify the cardiac concentrations of DNA, RNA, protein, the levels of activity of cardiac catalase and GSH-Px or the in vitro production of malonaldehyde of cardiac homogenates. Mitochondrial swelling at 24 hr and reduction of the mitochondrial numerical and volumetric densities along with myofilament fragmentation at 96 hr were the most significant ultrastructural changes in cardiac myocytes of ADR-treated rats. Although in vivo lipoperoxidation (diene conjugation) was detected in the cardiac lipids of only 2 out of 6 rats at 24 hr and in 3 out of 6 rats at 96 hr, no clear correlation could be found between the eventual presence of this in vivo phenomenon and any of the cardiac changes. These data suggested that lipoperoxidation may not play a fundamental role in the pathogenesis of acute ADR-cardiotoxicity in rats.

Immunocytochemical hormonal features of pituitary adenomas of aging Wistar male rats.

Pituitary adenomas were the most common grossly visible naturally occurring neoplasms found in over 27% of rats surviving beyond 17 months of age. Twenty-three pituitary adenomas, fixed in buffered neutral formalin and embedded in paraffin, were tested for the presence of prolactin (PRL), growth hormone (GH) and thyroid stimulating hormone (TSH) using the unlabeled peroxidase-antiperoxidase method. The adenoma cells in 6 (26%) of the 23 tumors stained with two or three of the tested hormones, but clear evidence that individual neoplastic cells contained more than one hormone was not found. These findings suggest that in aging male Wistar rats the spontaneous pituitary adenomas may originate from undifferentiated cells, PRL-, GH- and TSH-cells in a diminishing order of frequency.

Effects of the type of dietary fat at two levels of vitamin E in Wistar male rats during development and aging. III. Biochemical and morphometric parameters of the liver.

The purpose of this study was to explore in rats the possible influence of the type of dietary fat at two extreme levels of vitamin E on several biochemically determined hepatic changes and on a number of quantitatively analyzed structural and ultrastructural variations with age in hepatic cells. Six groups of weanling Wistar male rats were fed ad libitum isoenergetic diets containing similar amounts (15 g per 100 g diet) of saturated fat (coconut oil), unsaturated fat (safflower oil) or a combination of both at two levels of dl-alpha-tocopherol (2 or 200 mg per 100 g of diet). Determinations were performed in rats killed at 3, 6, 12, 18 and 24 months. Although in relation to age and irrespective of the type of diet, several of the biochemical parameters fluctuated with time, comparisons of the results between the youngest and oldest rats showed no changes in the levels of hepatic RNA, phospholipids, cholesterol, total tocopherols and total collagens, significant increases in DNA and triglycerides and a significant decrease in total protein. While the type of diet did not have in general significant influences on the levels of DNA, RNA, total protein and collagens, either the type of dietary fat and/or the levels of vitamin E had some definite effects on the levels of triglycerides, cholesterol, phospholipids and total tocopherols, as well as on the in vitro formation of malonaldehyde and on the eventual occurrence of in vivo lipoperoxidation (diene conjugation). These effects, however, varied in relation to the duration of the diverse dietary treatments. The morphologic studies indicated that all the livers had variable but generally moderate degrees of fatty changes (mainly due to triglyceride accumulation) which were attributed to the moderate obesity found in the rats. The mean nuclear and cell dimensions of hepatocytes, the number of binucleated hepatocytes, surface density of rough endoplasmic reticulum, numerical density of mitochondria and the fractional cytoplasmic volume occupied by lipofuscin pigment in hepatocytes were not significantly affected by the type of diet, by age or by the eventual occurrence of in vivo hepatic lipoperoxidation, whereas the numerical density of hepatocytes (mono- and binucleated) and "litoral cells" (endothelial, Kupffer and Ito cells), although unaffected by diet, significantly increased with age. On the other hand, the fractional volume of mitochondria and peroxisomes, as well as the numerical density of peroxisomes, were significantly influenced by the type of dietary fat and to lesser extent by the dietary levels of vitamin E.

Effects of the type of dietary fat at two levels of vitamin E in Wistar male rats during development and aging. I. Life span, serum biochemical parameters and pathological changes.

This experiment was designed to study in rats the implications of the dietary type of fat at two levels of vitamin E on the life span as well as on several biochemical and anatomopathological age-related changes. For this purpose, six different isoenergetic diets containing 15% coconut oil (SFD), safflower oil (UFD) or a combination of both (CFD) with 2 or 200 mg% of dl-alpha-tocopherol were offered ad libitum to outbred Wistar male rats from weaning to senescence. The results indicated that up to 9--12 months the body weights of rats consuming the CFD or the UFD increased generally faster than those fed the SFD, and that all rats developed moderate degrees of obesity. Age-dependent changes in organ weights (kidneys, testes, spleen, brain, liver and heart) were unaffected by diet. Serum levels of vitamin E generally reflected the corresponding dietary levels, but were also influenced by the type of dietary fat. Serum cholesterol levels were not significantly affected by the type of diet or by age. Only transient hypotriglyceridemic and hypophospholipidemic effects of the UFD were observed and, while the levels of triglycerides decreased with age up to the 18th month followed by an increase at 24 months, the levels of serum phospholipids remained unchanged. Neither diet nor age modified the serum albumin/globulin ratios. While no differences in maximum life span were found between dietary groups, the 50% survival time of rats fed the UFD at high level of vitamin E was significantly longer than in all the other groups. This beneficial effect was related to postponement of the onset and reduction of incidence of malignant neoplasms, but was apparently not related to any particular influence on the incidence or severity of chronic nephropathy which practically developed in all rats. Various neoplastic, degenerative and inflammatory diseases encountered in rats dying during the course of the experiment were tabulated and compared with similar findings reported by others in different strains of rats. Pituitary and adrenocortical adenomas as well as adrenocortical and renal carcinomas were the most frequent tumors found in this study. All the pathological changes provided useful baseline information for the evaluation of data presented in this and subsequent communications of this series of studies.