Tau and amyloid beta differentially affect the innate immune genes expression in Drosophila models of Alzheimer's disease and ß- D Mannuronic acid (M2000) modulates the dysregulation.

Alzheimer's disease (AD) is the most common cause of dementia and neuroinflammation is considered as one of the main culprits. The aim of this study was to evaluate the independent role of Aß42 and tau on the inflammatory pathway in the Drosophila models of AD and investigating the potential modulating effect of M2000 as a novel NSAIDs in those flies. The expression levels of relish, orthologs of NF-κB, antimicrobial peptide (AMP) including attacin A, diptericin B and a dual oxidase (Duox) as a ROS mediator, were evaluated in both M2000 treated and untreated groups followed by brain histology analysis to assess the extent of neurodegeneration. The potential inhibitory role of M2000 (ß-D Mannuronic acid) on the aggregation of tau protein was also investigated in vitro. According to the result, there was a significant induction of Duox, AMPs and its transcription factor expression in both aged and Drosophila models of AD which was in accordance with the increase in the number of vacuoles in the brain section of Drosophila models of AD. Interestingly M2000 treatment revealed a significant reduction in all neurodegeneration indexes; in vivo and anti-aggregating property; in vitro. Findings suggest that M2000 has potential to be an AD therapeutic agent.

Effects of Anethum graveolens L. on In Vitro Matured Mouse Oocytes and Granulosa Cells.

BACKGROUND: According to previous studies, Anethum graveolens L. (dill) aqueous extracts decreased the fertility of female rats. Therefore, the present study aimed to examine the effects of this herb on cultured granulosa cells and immature oocytes. METHODS: The cells were obtained from 27-29 day immature superovulated mice. The oocytes were cultured in a petri dish consisting of 30 µl drops of MEM-α and granulosa cells in a 24-well plate consisting of DMEM/F12 and different concentrations of 0, 10, 50, 100, 500, 1000, 10000 µg/ml of dill seed aqueous extract (DSAE) in 37°C and 5% CO2. Then, the in vitro maturation of oocytes, including Germinal Vesicle (GV), Germinal Vesicle Breakdown (GVBD), and meiosis II (MII) and oocyte bioviability were determined. Granulosa cells were then extracted and their bioviability, apoptosis, chromatin condensation, and lipid synthesis were examined. Estrogen and progesterone concentrations and Alkaline Phosphatase (ALP) activity were measured by RIA and spectrophotometry respectively from the supernatant of granulosa cell culture. RESULTS: The results revealed that concentration of 10000 µg/ml of DSAE were toxic and damaged granulosa cell growth and oocytes maturation. Lower concentrations were the same in the control group and did not have any side effects on cell growth. The number of lipid droplets, estrogen and progesterone concentrations, and ALP activity increased with higher doses of DSAE compared to those in the control culture. Additionally, apoptosis and chromatin condensation increased in higher concentrations of DSAE-(500 and 1000 µg/ml) treated cells. This herb extract decreased the oocytes maturation in dose-dependent manner. CONCLUSION: It was concluded that DSAE increased granulosa cells activity but damaged oocytes maturation, therefore it might be introduced as infertility agent.

The effects of Salvia officinalis L. on granulosa cells and in vitro maturation of oocytes in mice.

BACKGROUND: Salvia officinalisL. has been used since ancient times but there are little data about effects of this herb on normal reproductive cells. OBJECTIVE: To investigate the toxicity effects of Salvia officinalis L. on granulosa cells (GCs) and maturation of oocytes. MATERIALS AND METHODS: GCs and oocytes were extracted from superovulated ovaries of immature mice. The cells were treated with concentrations of 10, 50, 100, 500, and 1000 µg/ml of Salvia officinalis hydroalcoholic extracts and compared with the control culture. Bioviability, chromatin condensation, estradiol and progesterone concentrations, lipid synthesis, apoptosis, and alkaline phosphatase activity of GCs were measured. In vitro maturation of oocytes by determination of different maturation stages of oocytes including germinal vesicle, germinal vesicle breaks down, and metaphase II were examined. RESULTS: The results revealed that 500 and 1000 µg/ml concentrations of Salvia officinalisL. were toxic. The most of the GCs were in the early stages of apoptosis in 100 µg/ml treated culture and cell death happened with 500 µg/ml treatment. Progesterone concentration was reduced in 100 µg/ml and higher doses but estradiol concentration and alkaline phosphatase showed opposite effects. The lipid droplets content of GCs reduced significantly in all groups especially in 500 and 1000 µg/ml. Finally, oocyte's nucleus and cytoplasm showed a high level of condensation, and meiosis rate reduced in all treated cultures. CONCLUSION: Our findings suggested that higher dose of Salvia officinalis hydroalcoholic extracts inhibits, oocyte maturation, GCs bioviability, proliferation, and secretion.

Salvia officinalis L. induces alveolar bud growing in adult female rat mammary glands.

OBJECTIVES: In traditional medicine Salvia officinalis (sage) has been used as menstrual cycle regulator. In the present study the effects of sage extract on breast tissue were examined. MATERIALS AND METHODS: Fourteen female rats were divided into two groups: 1) Distilled water-treated rats (Con) that were gavaged with 1ml distilled water and 2) Saliva officinalis hydroalcoholic extract (SHE)-treated rats that were gavaged with 30mg/kg/body weight of sage extract for 30 days. The estrus cycle changes were monitored by daily examination of vaginal smear. Whole mounts of right pelvic breast were spread on the slide and stained by carmine. The number of alveolar buds (ABs) type 1 and 2 and lobules of mammary gland were scored. Tissue sections of left pelvic mammary gland were prepared and its histomorphometrical changes were measured. Blood samples were taken from dorsal aorta and estradiol and progesterone concentrations were measured using radioimmunoassay. RESULTS: Estrous cycles decreased significantly in SHE-treated animals. The number of alveolar buds and lobules in mammary gland whole mount of SHE-treated group were higher than the Con group. The number and diameter of ducts in histological section of mammary gland in SHE-treated group increased as compared to the Con group. CONCLUSION: Sage promotes alveologenesis of mammary glands and it can be used as a lactiferous herb.

Effects of Anethum graveolens L. (dill) on Oocyte and Fertility of Adult Female Rats.

BACKGROUND: Our previous studies revealed Anethum graveolens L. caused some changes in female reproductive system that induced infertility. Therefore, in this study, oocyte changes as one of probable reasons of infertility were investigated. METHODS: In this study, 59 adult female rats were divided into 3 groups of control, low dose (0.5 g/kg) and high dose (5 g/kg) of dill seed aqueous extract (LDE and HDE) treated groups that were gavaged with 1 ml of each dose for 10 days (2 estrous cycles). Vaginal smears were prepared daily. Oocytes of superovulated animals were extracted and their morphometrical changes were measured (n = 5). Oocyte cell membrane glycoconjugates were stained with UEA, PNA, and DBA-FITC lectins (n = 5). Ultrastructural studies of oocytes were performed using TEM (n = 5). The number, weight, and crown-rump length of newborns were examined in three groups after mating with untreated males (n = 5). Data were analyzed using SPSS software. RESULTS: Results demonstrated that the duration of the estrous cycle, the diestrus phase and progesterone concentration in the experimental groups increased significantly compared to the control group (p < 0.05). Granulosa cells of corpus luteum in HDE-treated group were larger and clearer. The intensity reactions of galactose/Nacetylgalactoseamine terminal sugar of oocyte decreased insignificantly in experimental groups compared to the control group p > 0.05. Duration of mating to pregnancy increased and the weight and crown-rump length of newborns decreased in experimental groups significantly (p < 0.05). CONCLUSION: Dill seed aqueous extract can induce infertility without any effect on oocyte structure.

Effects of plum extract on skeletal system of fetal and newborn mice.

OBJECTIVE: To evaluate the effects of Prunus domestica L. extracts on fetuses and neonatal skeletal systems. MATERIALS AND METHODS: A total of 32 pregnant mice (Mus musculus) received vehicle and plum hydroalcoholic extract at gestational days 1-18 and during the entire gestational period as well as 10 days postpartum, respectively. A total of 30 nonpregnant mice were fed plum hydroalcoholic extract and plum juice extract for 30 days. Bone calcium content and serum concentrations of calcium, magnesium and alkaline phosphatase were measured. The skeletal systems of their fetuses and neonates were stained with Alcian blue and alizarin red S and the length of femur, tibia, and their ossification center were measured. RESULTS: Crown-rump length of the newborn mice from mothers treated with plum extract (4.61 ± 0.25 mm) was higher compared to the control group (4.48 ± 0.31 mm, p = 0.001), and the femur osteogenesis index of newborn mice from mothers treated with plum extract was also higher (0.87 ± 0.09) compared to the control group (0.81 ± 0.06, p = 0.007). CONCLUSION: The findings showed that pregnant mice treated with plum extract had fetuses and newborn mice with higher osteogenesis index than those of the controls.

Mesenchymal stem cells repair germinal cells of seminiferous tubules of sterile rats.

BACKGROUND: Mesenchymal stem cells (MSCs) are undifferentiated cells that can differentiate and divide to other cell types. Transplantation of these cells to the different organs is used for curing various diseases. OBJECTIVE: The aim of this research was whether MSCs transplantation could treat the sterile testes. MATERIALS AND METHODS: In this experimental study, Donor MSCs were isolated from bone marrow of Wistar rats. The recipients were received 40 mg/kg of busulfan to stop endogenous spermatogenesis. The MSCs were injected into the left testes. Cell tracing was done by labeling the MSCs by 5-Bromo-2- Deoxy Uridine (BrdU). The immunohistochemical and morphometrical studies were performed to analysis the curing criteria. RESULTS: The number of spermatogonia (25.38±1.57), primary spermatocytes (55.41±1.62) and spermatozoids (4.95±1.30)×10(6) in busulfan treated animals were decreased significantly as compared to the control group (33.35±1.78, 64.44±2.00) and (10.50±1.82)×10(6) respectively but stem cells therapy help the spermatogenesis begin more effective in these animals (32.78±1.99, 63.59±2.01) and (9.81±1.33)×10(6) respectively than the control group. The injected BrdU labeled mesenchymal stem cells differentiated to spermatogonia and spermatozoa in the seminiferous tubules of the infertile testis and also to the interstitial cells between tubules. CONCLUSION: We concluded that testis of host infertile rats accepted transplanted MSCs. The transplanted MSCs could differentiate into germinal cells in testicular seminiferous tubules. This article extracted from M.Sc. Thesis. (Bentolhoda Fereydouni).

Effects of pomegranate extracts on cartilage, bone and mesenchymal cells of mouse fetuses.

Pomegranate is a rich source of polyphenols, which are believed to be responsible for the oestrogenic activities of extracts of this fruit in mice. One of these potential activities is the prevention of bone loss. The objectives of the present study were to determine the effects of pomegranate extract on chondrogenesis and osteogenesis in mouse embryos in vivo and limb bud cultures in vitro. A total of fifty pregnant Balb/c mice were given vehicle, pomegranate juice extract (PJE), pomegranate husk extract (PHE) or a mixture of husk and juice extract (PME). Their embryos were stained with alizarin red S and alcian blue, and the length of the femur, tibia and their ossification zones were measured on day 19 of gestation. Bone Ca content in pregnant mice was also measured. Mice treated with PJE showed an increase in bone Ca content. Dietary supplementation with all extracts significantly increased embryo femur length and osteogenesis index. Mesenchymal cells from fetal limb buds were cultured and exposed to 10, 100, 1000 and 10 000 µg/ml of PJE, PHE or PME. The number of viable cells was greater in cultures exposed to the extracts than in control cultures. The number of cartilage nodules and their diameters were greater in extract-treated cell cultures, a finding which reflected increased cell proliferation and differentiation rates. In conclusion, the findings of the present study suggest that pomegranate is able to enhance bone formation.

Effects of Anethum graveolens L. on fertility in male rats.

OBJECTIVES: The effects of Anethum graveolens seed extract on fertility of male rats were investigated. METHODS: Male Wistar rats were divided into five groups according to the treatment they received during 42 days: control, low dose (0.5 g/kg) and high dose (5 g/kg) of aqueous extracts, and low dose (0.045 g/kg) and high dose (0.45 g/kg) of ethanol extracts of Anethum graveolens seed. Sperm count and motility and testosterone concentration were measured. Sections of the testes, epididymis, and seminal vesicles were stained with peroxidase-conjugated lectins of Ulex europaeus agglutinin, peanut agglutinin, Dolichos biflorus agglutinin, soy bean agglutinin and concanavalin A. The treated male rats were mated with females and the crown-rump lengths and weights of their newborn pups were measured. RESULTS: No significant differences in sperm count, sperm motility or testosterone concentration were observed in the experimental groups. However, female rats did not become pregnant after mating with rats given the high dose of the ethanol extract. The distribution of terminal sugars on the epithelial surface of the reproductive structures decreased in the experimental groups. CONCLUSION: Anethum graveolens extract decreased fertility rate by modifying some terminal sugars on the cell surface of male reproductive organs involved in sperm maturation, capacitation and oocyte recognition.

Lectins influence chondrogenesis and osteogenesis in limb bud mesenchymal cells.

The role of cell surface glycoproteins in cell behavior can be characterized by their interactions with plant lectins. This study was designed to identify the effects of lectins on chondrogenesis and osteogenesis in limb bud mesenchymal cells in vitro. Limb bud mesenchymal cells from mouse embryos were cultured in high-density micromass culture. Wheat germ agglutinin (WGA), concanavalin A (ConA), peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA) and Ricinus communis agglutinin (RCA) were added separately to the culture media. Cells were cultured for 5 or 9 days, and cell viability was assayed by neutral red on day 5. The micromasses were stained with alcian blue, alizarin red S and Von Kossa stains, and alkaline phosphatase assays were also done. Dolichos biflorus agglutinin induced an increase in chondrogenesis, calcium precipitation and proteoglycan production. ConA and PNA did not affect chondrocyte differentiation but induced chondrocytes to produce more proteoglycan. Wheat germ agglutinin reduced chondrification and ossification but induced mesenchymal cells to store lipid droplets. Ricinus communis agglutinin 1 was toxic and significantly reduced cell survival. In conclusion, DBA was the most effective inducer of ossification and chondrification. Wheat germ agglutinin induced adipogenesis instead. These assays showed that lectins play important roles in limb bud development.

Cadmium-induced infertility in male mice.

The effects of cadmium in a concentration similar to that found in Maharloo Lake (Shiraz, Iran) on male reproductive system was studied in adult Balb/c male mice that received 0, 23, and 50 mg/kg of cadmium chloride in 0.5 mL distilled water for 45 days. Sperm count and motility, sperm nuclear maturity and chromatin structure tests were carried out. Testis of each mouse was examined histologically. The treated male mice were mated with females. Prostatic and nonprostatic acid phosphatase activity in blood serum, testis, and prostate, lipid peroxidation and cadmium accumulation in testis, seminal vesicle, and middle 1/3 of the quadericeps femoris muscle were measured. The sperm count, sperm motility, sperm maturity, and the level of testosterone decreased significantly in the high dose adminstered group. Histological studies showed a severe necrosis and atrophy in the testis of high dose group, consequently, there was no successful mating in some groups. The number of newborns and their weights and crown rump lengths reduced. Cadmium accumulation in testis and middle of the quadriceps femoris muscle was significantly higher in animals receiving 50 mg/kg cadmium chloride. Nonprostatic acid phosphatase activity decreased, whereas prostatic acid phosphatase activity increased significantly in serum of animals receiving 50 mg/kg cadmium chloride. Also prostatic acid phosphatase activity decreased significantly in prostate of animals receiving 50 mg/kg cadmium chloride. Lipid peroxidation was significantly higher in testis of animals treated with 50 mg/kg cadmium chloride compared with control group. Cadmium affects male reproductive system activity and can cause infertility in mice as an animal model.