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Abstract Background & aim: Aspartame is an artificial sweetener that has been used extensively in over 200 million people in more than 90 countries in various food products and pharmaceuticals. There are many controversial reports about the toxicity of aspartame on various tissues of the body. Therefore, the aim of the present study was to determine the effect of long-term aspartame on histomorphometric and histochemical adrenal glands in NMRI mice. Methods: In the present experimental study, 36 male NMRI male rats weighing 20-25grs were perched from Pasteur Institute of Experimental Animals. The rats were randomly divided into four groups: three groups received aspartate at dose of 40 (low dose of aspartame), 80 (medium dose of aspartame) and 160 (high dose of aspartame) mg/kg body weight by oral gavage for 90 days, respectively. And the control group was also considered. 24 hours after the last treatment, histological and histomorphometric changes were evaluated by digital microscopy. Also, specific stains of periodic acid schiff, trichrome masson and toluidine blue were used to determine carbohydrate compounds, amount of fibrosis and the number of mast cells in adrenocortical tissue. The collected data were analyzed by one-way ANOVA and Tukey's test. Results: Aspartate in the high dose group resulted in the collusion of cellular structure between glomerular cell and reticularis cells. Also, aspartame in the fashiculate region in medium to high dose groups caused cellular collusion and disintegration of the cellular columns and inflammatory and necrosis coronals were also observed in the feccilata region. In the central part of the adrenal gland, there were medium and high necrosis points in the adrenal gland. Histometric changes showed a significant increase in the size of the sponge cells, the number of mast cells, and the thickness of the layers of the fashiculate and reticularis. A significant reduction was observed in the large diameter of the central region of the adrenal gland in medium and high dose groups. No significant changes were seen in glomerular layer thickness and adrenal gland capsule parameters. In the histochemical studies of Trichromus Mason staining, it was found that aspartame increased the fibrosis tissue in the high dose group. Also, there was no significant change in the level of carbohydrate in the periodic acid staining in the groups. Conclusion: The results of this study revealed that aspartame as an oxidant in medium and high dose groups resulted in negative effects on histomorfrometry parameters and tissue damage in the adrenal gland by producing reactive oxygen species. It also increased the number of mast cells and fibrosis of the adrenocortical gland tissue.

Background and Objective: Soybean as a cheap protein and without side effects has been introduced to food industry. This study carried out to determine the effect of diet containing soybean on histology and histomorphometry of duodenal villi and serum levels of Calcium, Phosphorus and Glucose in mice. Methods: In this experimental study, 30 female immature BALB/c mice with 3 weeks of age were randomly allocated into control, and experimental 1 and 2. The control group was fed a diet with complete protein. Animals in the experimental 1 and 2 were received a diet of complete protein with 40% soybean and 20% soybean, respectively. After 3 months the mice were anesthetized and blood samples were taken from the heart for determining serum level of Calcium, Phosphorus and glucose. Duodenum specimens from were prepared and stained with hematoxylin and eosin. Results: Tissue structure of duodenum in experimental groups in compare to control group was not significantly changed, except for some scant specimens who showed some degrees of destruction in villi apicese. In histomorphometrical evaluation, the thickness of sub - mucosa and musculature were significantly increased in experimental groups compared to the controls group (P

Aspartame (ASP) is probably the best known artificial sugar substitute that is used widely in food. Many experimental studies have reported the toxicity of long-term administration of ASP in various organ tissues. However, there is little evidence available about the nature and mechanisms of the adverse effects of long-term consumption of ASP on the cardiovascular system. This study was conducted to evaluate the possible effects of ASP on heart tissue. For this study 36 mature male mice were divided into one control group and three groups which received respectively 40 mg/kg, 80 mg/kg and 160 mg/kg ASP orally, for 90 days. ASP at the doses of 80 and 160 mg/kg increased the serum content of malondialdehyde (MDA), but decreased serum nitric oxide (NO), creatine kinase (CK) and CK-MB, as well as blood superoxide dismutase (SOD) levels. Serum level of total anti-oxidant capacity (TAC) in blood was also reduced in serum at the dose of 80 mg/kg. Histochemical staining, including Periodic acid-Schiff, Masson's trichrome and Verhoeff-van Gieson staining, indicated that ASP at doses of 80 and 160 mg/kg reduced glycogen deposition and decreased the number of collagen and elastic fibres in the cardiac tissue. The cardiac expression of pro-apoptotic genes, including P53, Bax, Bcl-2 and Caspase-3, was modulated at the dose of 160 mg/kg. Moreover, transcription of Caspase-3 was up-regulated at the dose of 80 mg/kg. In conclusion, long-term consumption of ASP any higher than the acceptable daily intake (40 mg/kg) appears to act by promoting oxidative stress, has the potential to alter both histopathological and biochemical parameters, and induces P53-dependent apoptosis in cardiac tissue., (© 2022 Company of the International Journal of Experimental Pathology (CIJEP).)

Aspartame is one of the most common consumed artificial sweeteners utilized in many food products and beverages. It has been indicated that long-term consumption of aspartame leads to reproductive toxicity but its mechanism is not well-clear. In this study we investigated mechanism of aspartame-induced reproductive toxicity in male mice. For this purpose, 36 NMRI mature male mice received three doses of 40, 80, and 160 mg/kg body weight of aspartame, respectively per day by gavage for 90 days and also a control group was considered which received 0.5 mL of normal saline as the same route. The results revealed that long-term administration of aspartame at high doses significantly (P < .05) reduced gonadosomatic index, serum concentration of pituitary-testicular axis hormones (FSH, LH, and testosterone). It also decreased sperm parameters and total antioxidant capacity, antioxidant enzyme activities (superoxide dismutase, catalase, and glutathione peroxidase), while it caused increase in nitric oxide and malondialdehyde levels in testis tissue and sperm samples. Also, it decreased attenuated testicular histomorphometric indices (tubular differentiation index, spermiogenesis index, and repopulation index), and steroidogenic foci, while increased mRNA damages and apoptosis rate, downregulated antiapoptotic (Bcl-2) and upregulated proapoptotic (P53, BAX, and caspase-3) mediators respectively in testis. These findings indicated that consumption of aspartame for a long period results in male reproductive toxicity by decrease in serum concentration of pituitary-testis axis hormones and induction of oxidative stress and apoptosis in testis., (© 2020 Wiley Periodicals LLC.)

Background: Aspartame is one of the most commonly consumed artificial sweeteners that is widely used in foodstuffs. There are many debatable reports about aspartame toxicity in different tissues; however, on the subject of its effects on the reproductive system, few literatures are available. The present study was carried out for evaluating effects of aspartame on the reproductive system in male mice., Materials and Methods: In this experimental study, a total of 36 adult male mice were randomly divided into four groups of nine animals each. Three groups received aspartame at doses of 40, 80 and 160 mg/kg (gavage) for 90 days; also, a control group was considered. Twenty-four hours after the last treatment, animals were sacrificed. Then, body and testis weights, sperm parameters, serum testosterone concentration, total antioxidant capacity, and malondialdehyde (MDA) levels, antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px)] activities in blood, histomorphometrical indices and histochemical changes in testis were evaluated; also, mRNA and immunohistochemical expression of Hsp70-2 was measured in testis tissue., Results: The results revealed remarkable differences in sperm parameters, testosterone and oxidative stress biomarkers levels, and histomorphometrical indices, between the control and treatment groups. Also, in 80 and 160 mg/kg aspartametreated groups, expression of Hsp70-2 was decreased. Besides, in the aspartame receiving groups, some histochemical changes in testicular tissue were observed., Conclusion: The findings of the present study elucidated that long-term consumption of aspartame resulted in reproductive damages in male mice through induction of oxidative stress., Competing Interests: The authors declare no conflicts of interest., (Copyright© by Royan Institute. All rights reserved.)

This study described the morphological features of the Persian leopard (Panthera pardus saxicolor) tongue using light and scanning electron microscopy techniques. The keratinized filiform papillae were distributed all over the entire dorsal surface of the tongue and contained small processes. They were changed into a cylindrical shape in the body and conical shape in the root. The fungiform papillae were found on the apex and margin of the tongue. Few taste pores were observed on the dorsal surface of each papilla. The foliate papillae on the margins of the tongue were composed of several laminae and epithelial fissures. Taste buds were not seen within the non-keratinized epithelium. The vallate papillae were six in total and arranged in a "V" shape just rostral to the root. Each papilla was surrounded by a groove and pad. Taste buds were seen within their lateral walls. Lyssa was visible on the ventral surface of the tongue tip and was found as cartilaginous tissue surrounded by thin connective tissue fibres. The core of the tongue was composed of lingual glands, skeletal muscle and connective tissue. These glands were confined to the posterior portion of the tongue and were composed of many serous cells and a few mucous cells. The results of this study contributed to the knowledge of the morphological characteristics of the tongue of wild mammals and provided data for the comparison with other mammals., (© 2017 Blackwell Verlag GmbH.)

The present study made an attempt to measure the cortisol content, as an indicator of stress response, in rainbow trout embryos which were exposed to different densities and handling stress (air exposure) during incubation. The three densities of experimental embryos at early development stages were considered as 2.55 embryos/cm(2) (low density), 5.10 embryos/cm(2) (normal density) and 7.65 embryos/cm(2) (high density). The cortisol content of eggs (5.09 ± 0.12 ng/g) decreased to 3.68 ± 0.14 ng/g in newly fertilized eggs. Resting level of cortisol dropped at three densities by day 18 of post fertilization. Then, cortisol increased at hatching stage to 1.16 ± 0.11, 1.20 ± 0.12 and 1.21 ± 0.14 ng/g at low, normal and high densities, respectively. There were no statistically significant differences between cortisol concentrations in three densities. The acute handling stress test (5-min out-of-water), conducted on embryos (48 h post fertilization, organogenesis and eyed stage) in three densities, revealed no differences in whole-body cortisol levels between stressed and unstressed experimental groups. At hatching stage in low-density group, level of cortisol increased but the difference with the pre-stress levels was not statistically significant. Furthermore, significant differences in cortisol levels of stressed and unstressed embryos were detected on hatching in normal and high density groups [1.20 ± 0.12 at time 0-1.49 ± 0.11 ng/g at 1 hps (hours post stress) and from 1.21 ± 0.14 at time 0 to 1.53 ± 0.10 ng/g at 3 hps, respectively]. The results showed no difference in profile of cortisol in different densities, but acute stress conducted on embryos, incubated in different densities, revealed differences in cortisol stress response at hatching between normal and high density, which lead to cortisol increase at hatching time. It indicates that the lag time in the cortisol response to stressors immediately after hatching does not occur when the siblings were stressed during the embryo stage. Results, finally, indicated that hypothalamus-pituitary-interrenal axis was active and responded to an acute stressor under normal and high density, but it is unresponsive to a stressor around hatching under low density.

Background/aim: One of the most common psychiatric disorders in children is attention-deficit/hyperactivity disorder, which is treated extensively by methylphenidate. This study investigates the assessment of the effects of methylphenidate on histopathologic and histomorphometric changes of the testes and serum levels of gonadotropin in mice., Materials and Methods: In this study, 36 adult male mice were used. After determining their body weights, the animals were divided randomly into 2 experimental groups and 1 control group. The experimental groups received methylphenidate (2 and 10 mg kg(-1) day(-1)) via gavage for a period of 40 days. After evaluation of body weight, general anesthesia was used for taking blood samples from the heart in order to measure testosterone and levels of gonadotropin in serum. For the purpose of weighing the bodies and measuring the thickness of the germinal epithelium, the testes were removed and the possibility of any pathological changes was considered., Results: The results showed that methylphenidate could decrease the thickness of the germinal epithelium and body weight significantly, and could increase the levels of spermatogonia and serum gonadotropins and testosterone. Histopathological changes were also seen for vascular dilatation and congestion in interstitial tissue., Conclusion: Our findings demonstrated that administration of methylphenidate in adulthood may have an effect on spermatogenesis due to the influence of gonadotropin hormones on testis function.

Cortisol level changes in response to stocking density in the early stages of rainbow trout were measured. Eggs were exposed to low, normal, and high (2.55, 5.10 and 7.65 eggs cm(-2)) densities during the incubation period. Cortisol of maternal origin was found in pre-fertilized eggs (5.09 ± 0.12 ng g(-1)) of rainbow trout. In newly fertilized eggs, resting Cortisol levels (3.68 ± 0.14 ng g(-1)) decreased to 0.58 ± 0.08, 0.60 ± 0.12, and 0.57 ± 0.16 ng g(-1) at low, normal and high densities by day 10 (organo-genesis), respectively. Resting Cortisol levels remained constant until the eyed stage (day 18). Then, Cortisol showed an increase at hatching to 1.16 ± 0.11, 1.20 ± 0.12, and 1.21 ± 0.14 ng g(-1) at low, normal, and high densities, respectively. The pattern of change in Cortisol level was similar in all three densities. Interrenal cells were observed in 1-day old alevins at all three densities. Hematopoietic tissue, renal tubules and nucleated red blood cells were clarified through the head part of kidney. Higher numbers and larger interrenal cells were observed at high-density groups. Chronic density stress test conducted on embryonic stages of rainbow trout revealed no differences in Cortisol levels, but had an effect on the abundance and size of the interrenal cells. Densities were equaled after hatching (200 alevins per replicate) to investigate the different densities of eggs on stress indices in rainbow trout alevins. An acute stress (air exposure of eggs for five minutes) was applied in three treatments two weeks after hatching, and samples were taken at 0, 1, 3, 6, and 24 hps (hours post stress). Cortisol content increased under low density in 1 hps and reached from 5.21 ± 0.13 ng g(-1) to 6.01 ± 0.18 ng g(-1) (P < 0.05). Cortisol levels increased under normal density in 1 hps from 6.03 ± 0.28 ng g(-1) to 10.84 ± 0.18 ng g(-1) (P < 0.05). In high density also Cortisol increased from 6.83 ± 0.23 ng g(-1) to 8.86 ± 0.26 ng g(-1) (P < 0.05). At 3 hps;, Cortisol level was returned to basal level under low (P > 0.05) and normal (P > 0.05) densities, but significantly decreased (P < 0.05) under high density. Results revealed that the Cortisol biosynthesis was observed in rainbow trout between eyeing to the hatching stage. An increase in the density of eggs until 7.65 egg cm(-2) impaired Cortisol secretion and feedback system in alevins. However, more studies are needed to identify the exact time for Cortisol synthesis ability from eyeing to hatching in this species.

This article describes the histological and mucin histochemical properties of the small intestine of the Persian squirrel (Sciurus anomalus). This species is widely distributed in the Middle East and can be found as a companion animal. The histological studies revealed that the plicae circulares were not visible in the tunica mucosa. The maximum height and width of the villi were observed in the duodenum, which then decreased toward the ileum. The muscularis mucosa was scattered, whereas the tunica submucosa was composed of dense connective tissue. The lymphatic nodules were seen in the submucosa of the distal part of the jejunum and ileum, and Brunner's glands were embedded in the initial portion of the duodenum. The tunica muscularis was significantly thicker in the ileum, and the circular muscle layer was thicker than the longitudinal muscle layer throughout the entire length of the small intestine. The mucin histochemistry, which was examined using the periodic acid-Schiff (PAS) and alcian blue (AB) (pH 1.0 and 2.5) and also PAS-AB (pH 2.5) and aldehyde fuchsin-AB (pH 2.5) techniques coupled with methylation and saponification reaction for some sections, showed that the small intestine mucous content included both carboxylated and sulfated acidic mucins with few neutral mucins. The results of this study contribute to the knowledge of the histological and histochemical characteristics of the gastrointestinal tracts of exotic mammals and provide data for comparison with other mammals.

Phenol is a common industrial and ubiquitous environmental chemical which is used to synthesize resins and plastics. Due to its anesthetic and disinfectant properties, phenol is also widely used in pharmaceutical products. Since there were no adequate data about phenol immunotoxicity, the purpose of the present study is to investigate its toxic effects on the histological structures of the lymphoid organs in the mice. A total of 80 mice were randomly distributed into one control group and three experimental groups. The control group received only distilled water, whereas experimental groups were orally administered phenol at the concentrations of 80, 180, and 320 mg/kg/day, respectively. After 28 consecutive days, tissue samples were taken and histological changes of the spleens, thymuses, adrenal glands, and lymph nodes were examined using optical microscopy. The results showed that in the phenol treated animals; splenic megakaryocyte counts increased, the diameter of the splenic follicles decreased, the thymocyte population in both cortex and medulla reduced, the thickness of the reticular layers of adrenal gland increased and lymphatic cells populations in the lymph node were reduced, significantly ( P  < 0.01). Also, remarkable histological changes were noted in the various lymphatic organs of the treated mice. Overall, present findings give some histological evidences that selected qualitative and quantitative parameters of the lymphatic organs were significantly altered by phenol administration. In conclusion, the significant decreases of the immune cell populations together with histological alterations in the immunocompetent organs of the mice exposed to phenol indicate the immunosuppressive and immunotoxic properties of this chemical material.

Background: Due to common use of methylphenidate (MPH) for the treatment of Attention Deficit Hyperactivity Disorder (ADHD) and the role of the reproductive system in the production of gametes, studying the effects of this medication on the morphometry of testes, serum testosterone concentration, leydig cells function, and fertility rate was the aim of this study., Methods: Twenty seven male mice (Balb/C), eight weeks old, were randomly divided into one control and two treated groups. After weighing the mice, the treated groups received MPH (produced in Novartis company) at the doses of 2 mg/kg and 10 mg/kg for 40 days. The control group received only normal saline. Subsequently, after weighing the animals, the weights of testes, dimensions of the testis, and the serum testosterone concentration were measured in six mice belonging to each group. After tissue processing, the samples were stained with hematoxylin and eosin, then the leydig cells were counted. In order to assess male fertility in each group, 3 male mice were chosen and each of them was kept with three female mice in a separate cage. After 10 days, the fertility rates of the male mice were determined by counting the number of embryos in uterus and the corpora lutea in their ovaries., Results: The results of this study revealed that prescription of different doses of MPH can cause a significant decrease of the body weight. It reduces the number of leydig cells, too (p<0.01). Moreover, serum testosterone concentration (67.72±8.24 ng/ml in control group and 0.302±0.416 ng/ml after treatment with 2 mg/kg/day MPH) and fertility rate (95.42%±4.68% in control group and 64.96%±18.51% after treatment with 2 mg/kg/day MPH) of the male mice declined significantly in the treated groups compared with the control group (p<0.01), but it did not cause any changes in the weight or morphometric parameters of testes., Conclusion: The results of this study confirmed that MPH can negatively affect serum testosterone concentration and fertility rate of the male mice by decreasing the number of leydig cells and reducing the body weight.