Synergistic effects of pomegranate juice and atorvastatin for improving cerebellar structure and function of breast-feeding rats maternally fed on a high cholesterol diet.

A highly cholesterol-diet is associated with atherosclerosis and little about the development of cerebellar cortex disorder. The study illustrated the changes of cerebellar cortex of rat neonate maternally fed on high cholesterol diet and the capacity of pomegranate alone or in combination with atorvastatin to improve it. Eighty-eight pregnant Wister rats were divided into eight groups (n = 11); control, pomegranate supplemented group (daily orally 0.4 mL (20 %), atorvastatin (10 mg/kg BT), hypercholesterolemia (dietary consumption 3% cholesterol for 6 weeks prior to conception and throughout gestation and lactation period), hypercholesterolemia and pomegranate or atorvastatin, hypercholesterolemia and atorvastatin and pomegranate. Dams and their offspring were sacrificed at 21 days post-partum. Sera of mother and cerebellum of offspring were investigated biochemically as well as histo-cytological changes of cerebellar cortex of offspring. Offspring maternally fed on high cholesterol diet showed damage of the cerebellar Purkinje and granular cells associated with demyelination, increased caspase 3 immunohistochemistry and increased DNA damage. These were associated with decreased brain neurotransmitters and increase apoptic markers. Dams supplemented pomegranate and/or atorvastatin improved the assayed parameters more than that of atorvastatin alone. The authors concluded that pomegranate juice contains potent antioxidant nutrients capable of reducing the cytotoxicity of hypercholesterolemia and atorvastatin, and enhancing the structure and function of the cerebellar cortex.

Immunomodulatory effect of papaya (Carica papaya) pulp and seed extracts as a potential natural treatment for bacterial stress.

The current study evaluated the immunomodulatory effects of Carica papaya pulp and seeds methanol (MeOH) extracts on mice infected with Listeria monocytogenes. Gas chromatography-mass spectrometry analysis identified 10 active constituents in C. papaya seed MeOH extract and 10 compounds in C. papaya pulp MeOH extract. The experimental animals were divided into negative control (G1) group, positive control (G2) group, pulp extract treated (G3) group, and seed extract treated (G4) group. After infection of animals (G2, G3, and G4), treatments were started for 3 weeks. Estimation of the immunological parameters showed a marked decrease in IgM levels and an increase in IgG levels in the treated groups (G3 and G4) compared with those in G2. The proinflammatory cytokines (IL-10, IL-12, IL-1ß, IL-6, and TGF-ß1) were decreased in the treated groups (G3 and G4) compared with those in G2. Nitric oxide levels were also decreased, and the percentages of phagocytosis increased compared with those of G2. The results demonstrated the immunomodulatory and anti-inflammatory effects of C. papaya pulp and seeds MeOH extracts. PRACTICAL APPLICATIONS: Based on the antioxidant and antibacterial activities exhibited by the pulp and seed MeOH extracts investigated in this study, Carica papaya might be considered as a natural source of phytochemicals that could be utilized in novel foods and pharmaceuticals. Further investigation are needed to identify and purify compounds that might be responsible for the observed effects.

Anti-hypercholesterolemic impacts of barley and date palm fruits on the ovary of Wistar albino rats and their offspring.

A high cholesterol diet is related to ovarian dysfunction and infertility which has been increased among young ages consuming processed food products. The present study was conducted to evaluate the role of a high cholesterol diet on the ovaries of young female rats via assessments of histopathology, immunohistochemistry, oxidative stress and apoptic markers. Also, mating of hypercholesterolemic female rats was carried out to measure the fertility and numbers of their offspring. At the same time, phytotherapy was carried out through supplementing the diet with barley and/ or date palm fruits (10%) during the experiment to assess the phyto-therapeutic impacts in attenuation of drastic hypercholesterolemic effects. Hypercholesterolemic diet-fed rats exhibited damage of the ovarian follicles and increased follicular atresia. Furthermore, expression of cleaved caspase-3 was upregulated, while PCNA was downregulated in granulosa, theca and stroma cells. Hypercholesterolemic female rats showed marked depletion of antioxidative enzymes, increased lipid peroxidation and apoptotic markers. Alterations to the female serum hormones were detected. Offspring maternally fed on hypercholesterolemic diet showed a significant decrease of body weight and altered sex ratio. However, concomitant supplementation of barley and or date fruits to hypercholesterolemic groups revealed marked improvement of ovarian structure and function. On the basis of these evidences, it is believed that the enhanced synergistic effects of barley and/or date palm fruits in the amelioration of ovarian structure and functions were elicited by the potential antioxidant activity of their phytomicronutrients, polyphenols, ß-glucan and trace elements. These materials scavenge free radicals from inflamed cells that can be used to establish an effective and novel therapeutic strategy for activating ovarian cell regeneration.

Structural and functional abnormalities of hepatic tissues in male Wistar rats fed hyperwhey and super amino anabolic protein.

OBJECTIVE: Athletes and bodybuilders consume high-protein supplements to obtain energy and enhance the development and strength of their muscles. Over time, different investigations have revealed dysfunctions of their body organs. There are contradictions among scientists concerning the benefits and the alarm of developing body dysfunction. The aim of this study was to illustrate the effects on consumption of two anabolic protein supplements on body weight and structure and function of hepatocytes in male albino Wistar rats. METHODS: We assigned male Wistar albino rats into three groups (n = 10 each): control, hyperwhey protein (Nutrabolics, Richmond, Canada) (2.5 g/kg body weight), and super amino 2500 (SA) (APN, Ft. Launderale, FL, USA) (2.5 g/kg body weight). The applied dose was orally administered daily in tap water for 14 wk. Body weight was regularly measured. At 14 wk, animals were sacrificed and dissected. Blood was collected from a puncture of the heart and the liver was removed and weighed. Biochemical analysis of liver function tests, lipidogram, hematology, histopathology, transmission electron microscopy, immunohistochemistry of proliferating cell nuclear antigen, B-cell lymphoma 2 and 70 kd heat shock proteins, and flow-cytometry of hepatocyte cell cycle were performed. RESULTS: Hyperwhey- and SA-supplemented rats had lower body weight gain compared with the control group and developed hepatic dysfunction manifested by apparent congestion of blood vessel, increased apoptosis, and breakdown of hepatocytes. The SA group had thickening of the liver capsule and more drastic damage of hepatocytes. The level of transaminases was markedly increased. Insulin level was also markedly decreased in parallel with increase cholesterol, low-density lipoprotein, and triacylglycerols. CONCLUSION: Hyperwhey and SA protein formula administration dramatically altered the liver function and increased hepatic damage similar to the development of suspected diabetes.

Hypercholesterolemia-induced ocular disorder: Ameliorating role of phytotherapy.

The ocular region is a complex structure that allows conscious light perception and vision. It is of ecto-mesodermal origin. Cholesterol and polyunsaturated fatty acids are involved in retinal cell function; however, hypercholesterolemia and diabetes impair its function. Retinal damage, neovascularization, and cataracts are the main complications of cholesterol overload. Dietary supplementation of selected plant products can lead to the scavenging of free reactive oxygen species, thereby protecting the ocular regions from the damage of hypercholesterolemia. This review illustrates the dramatic effects of increased cholesterol levels on the ocular regions. The effect of phytotherapy is discussed in relation to the different regions of the eye, including the retina, cornea, and lens.

Cholesterol overload impairing cerebellar function: the promise of natural products.

The cerebellum is the part of the brain most involved in controlling motor and cognitive function. The surface becomes convoluted, forming folia that have a characteristic internal structure of three layers including molecular, Purkinje cell, and granular layer. This complex neural network gives rise to a massive signal-processing capability. Cholesterol is a major constituent, derived by de novo synthesis and the blood-brain barrier. Cholesterol is tightly regulated between neurons and glia-that is, astrocytes, microglia, and oligodendrocytes-and is essential for normal brain development. The axon is wrapped by myelin (cholesterol, phospholipids, and glycosphingolipids) and made up of membranes of oligodendrocytes, separated by periodic gaps in the myelin sheath, called nodes of Ranvier. Hypercholesterolemia is associated with increased oxidative stress and the development of neurotoxicity and Alzheimer's disease. Treatment with natural products has been found to support improved brain function and reduce low-density-lipoprotein cholesterol level. Fish oil is one such product; among the many plant products are: Morus alba leaves, fruit, and bark; pomegranate fruit and peel; Barley ß - glucans; date palm; and Allium sativum. The therapeutic potential was discussed in relation with the antilipidemic drugs, statins (HMG-CoA reductase inhibitors).

Effect of maternal diabetes and hypercholesterolemia on fetal liver of albino Wistar rats.

OBJECTIVE: The aim of this study was to predict the development of hepatic lesions and impairment of function during the development of fetuses (13-, 15-, 17-, and 19-d-old embryos) of diabetic and hypercholesterolemic mothers. METHODS: Eighty virgin and fertile male rats (one male/three females) of Wistar strain with an average body weight of 150 to 180 g were used. Mating was carried out, and pregnancy was determined by examining sperm in vaginal smears. Pregnant rats were arranged into three groups; control, diabetic (single intraperitoneal injection [i.p.] of 60 mg streptozotocin/kg) and hypercholesterolemic groups (fed on a diet containing 3% cholesterol for 6 wk before conception and throughout gestation) (n = 20). Pregnant rats were sacrificed and 13-, 15-, 17-, and 19-d-old embryos and livers were incised and subjected to histological and transmission electronic microscopical (TEM) investigations, assessments of alkaline phosphatase (Al-Pase) isoenzymes electrophoresis, DNA fragmentation, and comet assay. Flow cytometric analysis of apoptosis and caspases 3 and 9 in the livers of mother rats and their 19-d-old fetuses was determined. RESULTS: Histologic findings of diabetic and hypercholesterolemic mothers revealed apparent damage of hepatocytes, accumulation of lipid-laden cells, and vascular steatosis, while the 13-, 15-, 17- or 19-d-old fetuses of either diabetic or hypercholesterolemic mothers revealed disorganized hepatic architecture and massive cell damage. TEM of diseased mothers and their fetuses possessed increased incidence of pyknotic hepatocytes with massive vesicuolation of rough endoplasmic reticulum and degeneration of mitochondria. Al-Pase isoenzymes were altered and genomic DNA of both double and single helical structures were markedly damaged, especially in fetuses of maternally diabetic and hypercholesterolemic mothers. Flow cytometry revealed an increase in apoptosis and caspases 3 and 9 in diabetic and hypercholesterolemic mothers and their 19-d-old fetuses. CONCLUSION: These results suggested that maternal diabetes and hypercholesterolemia predicted early hepatitis and increased apoptosis in mothers and their fetuses as a result of oxidative stress and elevated apoptic markers caspases 3 and 9.

Analysis of fine structure and biochemical changes of retina during aging of Wistar albino rats.

BACKGROUND: Aging is a biological phenomenon that involves an increase of oxidative stress associated with gradual degradation of the structure and function of the retina. Gender differences and subsequent deterioration of retinal cell layers is an interesting topic, especially because there is no published work concerning it. METHODS: One hundred and twenty male and female Wistar albino rats ages 1, 6, 18, 30 and 42 months (n = 20 equal for male and female) were used. At the time interval, retinae were investigated by light and transmission electron microscopy, assessments of neurotransmitters, anti-oxidant enzymes (catalase, superoxide dismustase and glutathione S transferase), caspase-3 and -7, malonadialdhyde, and DNA fragmentation. RESULTS: Light and transmission electron microscopy observations of the older specimens (30 and 42 months) revealed apparent deterioration of retinal cell layers, especially ganglion and nerve fibres, nuclear, pigmented epithelium and stacked membranes of the photoreceptor's outer segments. Males were highly susceptible to aging processes. Retinal DNA fragmentation was remarked parallel with increase of apoptic markers caspase 3 and 7. Concomitantly, there was a marked reduction of neurotransmitters and anti-oxidant enzymes, and an increase of lipid peroxidation. CONCLUSIONS: Aging contributed to an increase of oxidative stress resulting from damage of mitochondria in retinal cells, a decrease of the anti-oxidant enzyme system and an increase of markers of retinal cell death.

Aging-related changes of optic nerve of Wistar albino rats.

Aging is a biological phenomenon that involves an increase of oxidative stress associated with gradual degradation of the structure and function of the optic nerve. Gender differences and subsequent deterioration of optic nerve are an interesting topic, especially because there is little published work concerning it. One hundred male and female Wistar albino rats' with ages 1, 6, 18, 24, and 30 months (n = 20 equal for male and female) were used. At the time interval, optic nerve was investigated by light and transmission electron microscopy (TEM), assessments of antioxidant enzymes (catalase, superoxide dismustase, and glutathione-S-transferase), caspase 3 and 7, malondialdhyde, flow cytometry of DNA, annexin v, and CD8, immunochemistry of vascular endothelial growth factor (VEGF), CD31, and CD45, and single-strand DNA fragmentation. Light and TEM observations of the older specimens (24 and 30 months) revealed apparent deterioration of optic nerve axons, abundant oligodendrocytes with pyknotic nuclei, swollen astrocytes, angiogenesis, vacuolar degeneration, and mitochondrial damage. Females were highly susceptible to aging processes. Concomitantly, there was a marked reduction of antioxidant's enzymes and an increase of lipid peroxidation and apoptotic markers. Old age exhibited a marked increase of G1 apoptosis, UR and LR of annexin V and CD8 as well as increased immuno-positive reaction with VEGR, CD31 and CD45. We conclude that aging contributed to an increase of oxidative stress resulting from damage of mitochondria in axons, oligodendrocytes, and astrocytes. Age-related loss of optic nerve axons is associated with multifactorial agents including reduction in antioxidant enzymes, disruption of vasculature, astrocyte, and oligodendrocyte, demyelination, and damage of mitochondria, which enhance the liberation of reactive oxygen species as assessed by an increase of apoptotic markers malondialdhyde and caspase 3 and 7.

Aging related changes of retina and optic nerve of Uromastyx aegyptia and Falco tinnunculus.

Aging is a biological phenomenon that involves gradual degradation of the structure and function of the retina and optic nerve. To our knowledge, little is known about the aging-related ocular cell loss in avian (Falco tinnunculus) and reptilian species (Uromastyx aegyptia). A selected 90 animals of pup, middle, and old age U. aegyptia (reptilian) and F. tinnunculus (avian) were used. The retinae and optic nerves were investigated by light and transmission electron microscopy (TEM) and assessments of neurotransmitters, antioxidant enzymes (catalase, superoxide dismustase and glutathione s transferase), caspase-3 and -7, malonadialdhyde, and DNA fragmentation. Light and TEM observations of the senile specimens revealed apparent deterioration of retinal cell layers, especially the pigmented epithelium and photoreceptor outer segments. Their inclusions of melanin were replaced by lipofuscins. Also, vacuolar degeneration and demyelination of the optic nerve axons were detected. Concomitantly, there was a marked increase of oxidative stress involved reduction of neurotransmitters and antioxidant enzymes and an increase of lipid peroxidation, caspase-3 and -7, subG0/G1 apoptosis, and P53. We conclude that aging showed an inverse relationship with the neurotransmitters and antioxidant enzymes and a linear relationship of caspases, malondialdhyde, DNA apoptosis, and P53 markers of cell death. These markers reflected the retinal cytological alterations and lipofuscin accumulation within inner segments.

Cardiomyopathy and angiogenesis defects of Wistar rat fetuses of diabetic and hypercholesterolemic mothers.

OBJECTIVE: We aimed to illustrate the histogenesis, lactic dehydrogenase isoenzymes electrophoresis, and DNA damage of cardiac muscles and blood vessels during prenatal life of maternal diabetic or hypercholesterolemic mother. METHODS: Eighty fertile male and virgin female Wistar rats (1 male/3 females), weighing approximately 130 g, were mated and zero date of gestation was determined. Diabetes was induced at the fifth day of gestation by intraperitoneal injection of a single dose of 60 mg streptozotocin/kg body weight in citrate buffer, pH 4.6. At the same time, hypercholesterolemia was carried out by feeding virgin rats a diet containing 3% cholesterol for 6 wk before the onset of conception. Pregnant rats were arranged into three groups: control, diabetic, and hypercholesterolemic (n = 20). The animals were sacrificed and embryos were separated at 7-, 13-, 15-, 17-, and 19 d old, respectively, and subjected to light and transmission electron microscopy, lactic dehydrogenases isoenzymes electrophoresis, DNA fragmentation, and comet assay. The sera of the mothers were examined for fasting glucose level, total cholesterol, triglycerides, low-density lipoprotein, high-density lipoprotein, and creatine phosphokinase levels. RESULTS: Diabetic and hypercholesterolemic mothers exhibited a significant increase of sera cholesterol level, low-density lipoprotein, and creatine phosphokinase activity. Histologic findings of embryos of diabetic and hypercholesterolemic mothers revealed cardiomyopathy and malformation of blood vessels with an apparent degeneration of their endothelium. Transmission electron microscopy possessed massive necrosis of muscle fibers, disorganization of Z and I bands, and mitochondrial damage. Lactic dehydrogenase isoenzyme electrophoresis was altered and genomic DNA fragmentation was markedly increased. CONCLUSION: Maternal diabetes or hypercholesterolemia led to marked alterations in blood vessel differentiation as well as to cardiomyopathy during prenatal growth as assessed by the disruption of fine structures, abnormal lactic dehydrogenase isoenzymes electrophoresis, and an increase of DNA damage. These may be attributed to the marked oxidative stress and liberation of free oxygen radicals, which interrupted the myocardium structure and function during organogenesis.

Studies on tongue of reptilian species Psammophis sibilans, Tarentola annularis and Crocodylus niloticus / Estudios sobre la lengua de las especies de reptiles Psammophis sibilans, Tarentola annularis y Crocodylus niloticus

Three different reptilian species Psammophis sibilans (Order Ophidia), Tarentola annularis (Order Squamata and Crocodylus niloticus (Order Crocodylia) are used in the present study. Their tongue is removed and examined morphologically. Their lingual mucosa examined under scanning electron microscopy (SEM) as well as processed for histological investigation. Gross morphological studies revealed variations of tongue gross structure being elongated with bifurcated end in P. sibilans or triangular flattened structure with broad base and conical free border in T. annularis or rough triangular fill almost the floor cavity in C. niloticus. At SEM, the lingual mucosa showed fine striated grooves radially arranged in oblique extension with missing of lingual papillae. Numerous microridges are detected above the cell surfaces in P. sibilans. T. annularis exhibited arrangement of conical flattened filiform papillae and abundant of microridges. However in C. niloticus, the lingual mucosa possessed different kinds of filiform papillae besides gustatory papillae and widespread arrangement of taste buds. Histologically, confirmed SEM of illustrating the lingual mucosa protrusion of stratified squamous epithelium in P. sibilans and presence of apical taste buds in conical filiform papillae of T. annularis. The lingual mucosa of C. niloticus revealed different forms of both filiform and gustatory papillae.

En el presente estudio fueron utilizados tres diferentes especies de reptiles Psammophis sibilans (Orden Ophidia), Tarentola annularis (Orden Squamata y Crocodylus niloticus (Orden Crocodylia). Sus lenguas fueron removidas y examinadas morfológicamente. Las mucosas linguales se examinaron bajo microscopio electrónico de barrido (SEM) y también procesadas para la investigación histológica. El estudio morfológico macroscópico reveló variaciones de la estructura de la lengua, siendo éstas alargadas con el extremo bifurcado en P. sibilans o de estructura triangular aplanada con una base amplia y el borde libre cónico en T. annularis o triangular rugosa llenando casi todo el suelo de la cavidad en C. niloticus. En el SEM, la mucosa lingual mostró finas ranuras estriadas dispuestas radialmente en la extensión oblicua con pérdida de las papilas linguales. Numerosas microcrestas fueron detectadas por encima de la superficie de las células en P. sibilans. T. annularis exhibiendo una disposición de papilas filiformes cónicas aplanadas y abundante de microcrestas. Sin embargo, en C. niloticus, la mucosa lingual posee diferentes tipos de papilas filiformes además de papilas gustativas con una extensa disposición de corpúsculos gustativas. Histológicamente, mediante confirmación de SEM se ilustra la protrusión del epitelio escamoso estratificado de la mucosa lingual en P. sibilans y la presencia de corpúsculos gustativos apicales en las papilas filiformes cónicas de T. annularis. La mucosa lingual de C. niloticus reveló diferentes formas de ambas papilas, filiformes y gustativas.