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This study aimed at investigating the relationship between concentrations of macro and trace elements in blood serum, and fluids from small and large follicles (SFF and LFF, respectively), oviduct (OF), and uterus (UF) of female dromedary camels. Fluids from small (2-6 mm) and large follicles (7-20 mm), oviduct and uterus, and blood samples were collected from 19 camels. The results indicated that the concentrations of serum Mg, Fe, and Mn were significantly higher than their follicular fluid, OF, and UF concentrations. Levels of Zn, Fe, Cu, Cr, and Mn were significantly higher in SFF than in LFF. Se and Mo concentrations were higher in LFF. Co concentration was lower in serum than in reproductive tract fluids. Cr concentration was higher in UF and OF than in the serum, SFF, and LFF. High Ca concentration was observed for serum and SFF, followed by LFF. The concentration of Na was about 1.18-fold higher in SFF than in serum, OF, and LFF, and approximately 4.1-fold higher in serum than in UF. K was present in higher concentration in SFF than in serum and LFF; however, its concentration was low in UF and OF. In conclusion, this study shows the concentrations of certain elements in small and large follicular, uterine, and oviductal fluids, which may be low or high depending on their function in the development and growth of follicles. This information can support the development of new media for in vitro oocyte maturation and fertilization of female camels.
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Camelus/metabolismo , Líquido Extracelular/química , Oviductos/química , Oligoelementos/metabolismo , Útero/química , Animais , Camelus/sangue , Feminino , Líquido Folicular/química , Oligoelementos/sangueRESUMO
All-trans retinoic acid (RA) is a metabolite of vitamin A and has pleiotropic actions on many different biological processes, including cell growth and differentiation, and is involved in different aspects of fertility and developmental biology. In the current study, we investigated the effects of RA on camel (Camelus dromedarius) cumulus-oocyte complex in vitro maturation (IVM). IVM medium was supplemented with 0, 10, 20, and 40 µM RA. Application of 20 µM RA significantly reduced the proportion of degenerated oocytes and significantly improved oocyte meiosis and first polar body extrusion compared to the control and other experimental groups. Retinoic acid significantly reduced the mRNA transcript levels of apoptosis-related genes, including BAX and P53, and reduced the BAX/BCL2 ratio. In addition, RA significantly reduced the expression of the Transforming growth factor beta (TGFß) pathway-related transcripts associated with the actin cytoskeleton, ACTA2 and TAGLN; however, RA increased TGFß expression in cumulus cells. The small molecule SB-431542 inhibits the TGFß pathway by inhibiting the activity of activin receptor-like kinases (ALK-4, ALK-5, and ALK-7); however, combined supplementation with RA during IVM compensated for the inhibitory effect of SB-431542 on cumulus expansion, oocyte meiosis I, and first polar body extrusion in activated oocytes. The current study shows the beneficial effects of RA on camel oocyte IVM and provides a model to study the multifunctional mechanisms involved in cumulus expansion and oocyte meiosis, particularly those involved in the TGFß pathway.
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Células do Cúmulo/citologia , Técnicas de Maturação in Vitro de Oócitos , Oócitos/citologia , Tretinoína/farmacologia , Animais , Apoptose/efeitos dos fármacos , Benzamidas/farmacologia , Blastocisto/citologia , Camelus , Meios de Cultura , Células do Cúmulo/efeitos dos fármacos , Dioxóis/farmacologia , Técnicas de Cultura Embrionária , Feminino , Fertilidade , Técnicas de Transferência Nuclear , Oócitos/efeitos dos fármacos , Fator de Crescimento Transformador beta/metabolismoRESUMO
This study was conducted using 120 multiparous Awassi ewes during the breeding season to compare the effects of using previously used controlled internal drug release (CIDR) on the hormone profiles, reproductive performance and economic measures of ewes. Ewes were randomized to receive one of five previously used CIDR (previously used for 6, 12, 18, 24 or 30 days) or the new CIDR as a control for 6 days (CIDR6, CIDR12, CIDR18, CIDR24, CIDR30, and CIDR0 [control], respectively). Blood samples were collected on four occasions, at the time of CIDR insertion, after 3 days of insertion, and at the time of withdrawal and insemination. Serum estradiol (E2) and progesterone (P4) concentrations were measured. Timed insemination was performed 48 hr post-CIDR withdrawal. Pregnancy was diagnosed by ultrasonography 23 days after insemination and confirmed on day 35. The heat detection rate was significantly (p < 0.05) higher in the CIDR0 and CIDR6 groups than in the CIDR18 and CIDR30 groups. The total pregnancy rate and fecundity were significantly (p < 0.05) higher in the CIDR6 group than in other groups. P4 level was significantly (p < 0.05) higher in the CIDR0 group than in the CIDR30 group at the time of removal. At each time point, the E2 level was significantly (p < 0.05) higher in the CIDR6 group than at the other groups. The total variable cost, total cost, return and net profit were higher in the CIDR6 and CIDR0 groups than in the other groups. In conclusion, although previously used CIDRs are efficient at synchronizing oestrus in ewes, the duration of previously usage significantly affected the reproductive parameters and economic profit. CIDRs previously used for 6 days and new CIDRs provided the highest fertility and fecundity rates, besides return and net profit. Economically, it is not advisable to use CIDRs that previously used for 12 days or more.
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Preparações de Ação Retardada/administração & dosagem , Sincronização do Estro/efeitos dos fármacos , Inseminação Artificial/economia , Progesterona/administração & dosagem , Animais , Feminino , Fertilidade/efeitos dos fármacos , Inseminação Artificial/veterinária , Gravidez , Taxa de Gravidez , OvinosRESUMO
The dromedary camel (Camel dromedarius) is physiologically well adapted to life in hot, dry and barren land. In the present study, we report the tolerance of camel oocytes and cumulus cells to acute and chronic heat shock. Camel oocytes and cumulus cells were exposed to acute (45⯰C for 2â¯h) and chronic (45⯰C for 20â¯h) heat shock. Our results demonstrated that acute and chronic heat shock altered malondialdehyde concentration, which is a marker for oxidative stress. Furthermore, the heat shock reduced glutathione levels during in vitro oocyte maturation. The expression of two well-known heat shock proteins HSP70 and HSP90 were increased similarly in oocytes and cumulus cells after acute heat shock. Oocytes were less tolerant to the short acute heat shock, and showed decreased maturation, which leads to reduction in ooplasmic diameter and an increase in chromosomal count abnormalities. Furthermore, the pro-apoptotic genes P53 and BAX had increased expression levels, whereas for the anti-apoptotic gene such as BCL2 expression levels was decreased. On the other hand, the cumulus cells tolerated acute and chronic heat shock, as evident by the increase in HSP70 and HSP90 expression and steady expression levels of P53, BAX, and BCL2 after acute hyperthermia. Cumulus cells regained their vitality and ability to proliferate after chronic hyperthermia and showed wound healing capabilities after 9 days of chronic hyperthermia. Collectively, these results indicate the adaptive tolerance of camel somatic cells to acute and chronic heat shock, which is lethal to cells in many other mammals.
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Camelus/metabolismo , Células do Cúmulo/metabolismo , Febre/metabolismo , Resposta ao Choque Térmico , Oócitos/metabolismo , Animais , Apoptose , Proliferação de Células , Células Cultivadas , Feminino , Proteínas de Choque Térmico HSP72/metabolismo , Proteínas de Choque Térmico HSP90/metabolismo , Peroxidação de LipídeosRESUMO
IMPORTANCE: The creation of robust maternal-embryonic interactions and implantation models is important for comprehending the early stages of embryonic development and reproductive disorders. Traditional two-dimensional (2D) cell culture systems often fail to accurately mimic the highly complex in vivo conditions. The employment of three-dimensional (3D) organoids has emerged as a promising strategy to overcome these limitations in recent years. The advancements in the field of organoid technology have opened new avenues for studying the physiology and diseases affecting female reproductive tract. OBSERVATIONS: This review summarizes the current strategies and advancements in the field of 3D organoids to establish maternal-embryonic interaction and implantation models for use in research and personalized medicine in assisted reproductive technology. The concepts of endometrial organoids, menstrual blood flow organoids, placental trophoblast organoids, stem cell-derived blastoids, and in vitro-generated embryo models are discussed in detail. We show the incorportaion of organoid systems and microfluidic technology to enhance tissue performance and precise management of the cellular surroundings. CONCLUSIONS AND RELEVANCE: This review provides insights into the future direction of modeling maternal-embryonic interaction research and its combination with other powerful technologies to interfere with this dialogue either by promoting or hindering it for improving fertility or methods for contraception, respectively. The merging of organoid systems with microfluidics facilitates the creation of sophisticated and functional organoid models, enhancing insights into organ development, disease mechanisms, and personalized medical investigations.
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Organoides , Feminino , Animais , Gravidez , Humanos , Técnicas de Cultura de Células em Três Dimensões/métodos , Implantação do Embrião/fisiologiaRESUMO
Introduction: This research aimed to examine the effects of water-added betaine (BET) and/or nano-emulsified vegetable oil (MAGO) on carcass and meat quality characteristics of broilers raised under thermoneutral (TN) and heat stress (HS) conditions. Methods: On day 21, 640 birds (Ross 308) were randomly assigned to one of two thermal conditions (thermoneutral 22 ± 1°C and heat stress 32 ± 1°C) each containing four treatment groups: Control, BET, MAGO, and a mixture of both (BETMAGO) in a 2 × 4 factorial arrangement (eight groups). Each group has eight replicates, with ten birds each. The birds' carcass and meat quality characteristics were evaluated at 35 days. Results and discussion: The dressing percentage, breast, leg, wing, heart, initial pH, color change, cooking loss (CL), water-holding capacity (WHC), shear force (SF), and texture profile with exception of springiness significantly affected by the treatments. The results showed that HS had negative effects on carcass weight and relative weights of the breast, spleen, and heart. Moreover, HS increased dressing percentage, wing, initial pH, final core temperature, initial lightness, WHC, and hardness. Significant differences in interactions between treatments and temperature were observed in the spleen, WHC, and SF. Conclusion: Water supplemented with BET effectively improved carcass dressing percentage, breast weight, and meat quality in terms of water-holding capacity and tenderness under HS conditions. More studies on the use of BET and/or MAGO at different levels were recommended.
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This is the first report on a biphasic in vitro maturation (IVM) approach with a meiotic inhibitor to improve dromedary camel IVM. Spontaneous meiotic resumption poses a major setback for in vitro matured oocytes. The overall objective of this study was to improve in vitro maturation of dromedary camel oocytes using ROCK inhibitor (Y-27632) in a biphasic IVM to prevent spontaneous meiotic resumption. In the first experiment, we cultured immature cumulus-oocyte complexes (COCs, n = 375) in a prematuration medium supplemented with ROCK inhibitor (RI) for 2 h, 4 h, 6 h, and 24 h before submission to normal in vitro maturation to complete 28 h. The control was cultured for 28 h in the absence of RI. In the first phase of experiment two, we cultured COCs (n = 480) in the presence or absence (control) of RI for 2 h, 4 h, 6 h, and 24 h, and conducted real-time relative quantitative PCR (qPCR) on selected mRNA transcripts. The same was done in the second phase, but qPCR was done after completion of normal IVM. Assessment of nuclear maturation showed that pre-IVM for 4 h yielded an increase in MII oocyte (54.67% vs. 26.6% of control; p < 0.05). As expected, the same group showed the highest degree (2) of cumulus expansion. In experiment 2, qPCR results showed significantly higher expression of ACTB and BCL2 in the RI group treated for 4 h when compared with the other groups. However, their relative quantification after biphasic IVM did not reveal any significant difference, except for the positive response of BCL2 and BAX/BCL2 ratio after 4 and 6 h biphasic IVM. In conclusion, RI prevents premature oocyte maturation and gave a significantly positive outcome during the 4 h treatment. This finding is a paradigm for future investigation on dromedary camel biphasic IVM and for improving the outcome of IVM in this species.
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This study was conducted to compare the effects of acute hyperthermia (45 °C for 4 h) on the viability, proliferation, and migratory activity through wound-healing assays of cow and sheep fibroblasts. The study examined the effects on primary cultures and first passage skin-derived fibroblasts. Relative quantification of HSP70, HSP90, P53, BAX, BCL2, and BECN1 was investigated after normalization to housekeeping genes GAPDH and beta-actin. The results revealed that cultured cow primary fibroblasts exhibited increased viability and reinitiated cell migration to close the cell monolayer scratch earlier than sheep cells. Similar patterns were observed in the first passage fibroblasts, with severe effects on sheep cells. Both cow and sheep cells exhibited decreased cell viability and failed to regain migratory activity after re-exposure of recovered heat-shocked cells. Effects of hyperthermia on sheep cells were potentiated by cell cryopreservation. The qPCR results showed that cow cells significantly increased HSP70 and HSP90 expression, which decreased the elevation of P53, and ameliorated the effects of the increased BAX/BCL2 ratio. The results provide a paradigm to compare thermotolerance among different animal species and revealed that trypsin could be an additional stress, which potentiates the effects of heat shock in in vitro experiments.
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There are several obstacles in camel semen cryopreservation; such as increasing semen viscosity and the reduction in motile spermatozoa after ejaculation. Epididymal spermatozoa offer an efficient alternative to overcome these problems and are well-suited for artificial insemination in camels. In the current study, we compared the effects of supplementation with vitamin C, E, inorganic trace elements of selenium (Na2SeO3) and zinc (ZnSO4), and zinc and selenium nanoparticles (ZnONPs and SeNPs, respectively) on the cryopreservation of dromedary camel epididymal spermatozoa. When the SHOTOR extender was supplemented with ZnONPs and SeNPs; the sperm showed increased progressive motility; vitality; and membrane integrity after cooling at 5 °C for 2 h; when compared to the control and vitamin-supplemented groups. Moreover, the ZnONPs and SeNPs supplementation improved the progressive motility, vitality, sperm membrane integrity, ultrastructural morphology, and decreased apoptosis when frozen and thawed. SeNPs significantly increased reduced glutathione (GSH), superoxide dismutase (SOD), and decreased lipid peroxide malondialdehyde (MDA) levels. The advantageous effects of the trace elements were potentiated by reduction into a nano-sized particle, which could increase bioavailability and reduce the undesired liberation of toxic concentrations. We recommend the inclusion of SeNPs or ZnONPs to SHOTOR extenders to improve the cryotolerance of camel epididymal spermatozoa.
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Oocyte vitrification preserves the female genetic resources of elite dromedary camels. In the current study, we aimed to explore the effects of vitrification of camel oocytes on mitochondrial activity, redox stress, and expression of genes related to mitochondrial function, apoptosis, pluripotency, and cytoskeleton. Moreover, we investigated developmental competence of vitrified oocytes after parthenogenetic activation. Oocytes vitrified with the Cryotop method were compared with the fresh oocytes. Our results showed that vitrification led to increased ROS production in oocytes as evidenced by an increase in the DCFDHA fluorescence intensity, and lower mitochondrial activity. At the molecular level, vitrification reduced mRNA expression of many genes, including those related to mitochondrial function (TFAM, MT-CO1, MFN1, ATP1A1, NRF1), pluripotency (SOX2 and POU5F1), and apoptosis (p53 and BAX). In contrast, expression of KLF4 and cytoskeleton-related genes (ACTB and KRT8) was not affected. However, we found no difference in the rates of oocyte survival, cleavage, and blastocyst development, and blastocyst hatching between fresh and vitrified oocytes after warming. Our results indicate that although vitrification of camel metaphase II (MII) oocytes adversely affected mitochondrial functions, the effect was transient without compromising the developmental potential of the oocytes after parthenogenetic activation.
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Camelus , Vitrificação , Animais , Criopreservação , Feminino , Mitocôndrias , Oócitos/metabolismo , Injeções de Esperma IntracitoplásmicasRESUMO
The aim of the study was to investigate the effects of different concentrations of resveratrol on head morphology, motility characteristics, oxidative state and in vitro fertility of cooled ram spermatozoa. Pooled semen from three Najdi rams was diluted with Triladyl® having different concentrations of resveratrol, zero (control), 200 µM (45.65 µg/mL) and 400 µM (91.30 µg/mL) resveratrol, then stored at 5 °C for 168 h. The head morphometric, sperm kinematic parameters, Malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD) and in vitro fertilizing capability of ram spermatozoa were evaluated after 24, 72, 120 and 168 h of cooling storage. The total motility (TM) of the sperm with resveratrol at 200 µM and 400 µM was significantly (p ≤ 0.05) higher than that in the control group at 72 and 120 h of cooling storage. On the other hand, the progressive motility (PM) of the sperm with resveratrol at 200 µM and 400 µM was significantly (p ≤ 0.05) higher than that in the control group at 168 h of cooling storage period. After 168 h of cooling storage, significantly higher straightness (STR) was observed in 400 µM group than two other groups and in 200 µM group than the control group. Both resveratrol groups had higher linearity (LIN) than control one at 120 and 168 h of cooling storage. The length, width and area of sperm head were lower (P ≤ 0.05) in the control compared to the other treatment groups after 120 and 168 h of storage. There was a significant increase in superoxide dismutase (SOD) concentration in the two resveratrol groups compared with the control one over the seven days of cooling storage and the same result was found in the reduced glutathione (GSH) concentration at 24, 72, and 168 h of storage. There was a significant (p ≤ 0.05) decrease in malondialdehyde (MDA) concentration in the 400 µM resveratrol group than that in two other groups over the seven days of storage period. Cleavage and blastocyst rates following in vitro fertilization were significantly higher (p ≤ 0.05) in 400 µM resveratrol than other groups at 72 h for cooling storage period. In conclusion, addition of resveratrol in the extender can protect sperm head morphology, improve kinematic parameters and in vitro fertility, and reduce oxidative stress of ram spermatozoa during liquid storage at 5 °C.
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Antioxidantes/farmacologia , Resveratrol/farmacologia , Preservação do Sêmen/veterinária , Sêmen/efeitos dos fármacos , Ovinos/fisiologia , Animais , Blastocisto , Fertilidade , Temperatura , Fatores de TempoRESUMO
This study examined the effect of supplemental betaine on live performance, selected blood parameters, and gene expression of water channel proteins (Aquaporins, AQP) of broiler chicks delayed in placement for 48 h post-hatch. In total, 540 newly-hatched male broiler chicks were obtained from a local hatchery and were randomly allotted to one of five treatments with nine replicates per treatment (12 chicks per replicate). Chicks were either placed immediately, control; held for 48 h post-hatch with no access to feed or water, Holdnull; held for 48 h with free access to drinking water only, HoldW; held for 48 h with free access to drinking water supplemented with 1 ml per L of betaine solution (40% betaine), HoldB1; or held for 48 h with free access to drinking water supplemented with 2 ml per L of betaine solution (40% betaine), HoldB2 group. The results showed that post-hatch holding for 48 h depressed feed intake and body weight gain during the entire 15 d study period with no beneficial effect of supplemental betaine. Chicks in the HoldB2 group had elevated serum glucose, triglycerides, and aspartate aminotransferase 48 h post-hatch. Early water deprivation directly affected the brain proopiomelanocortin (POMC) and hepatic glucocorticoid receptors (GR) expression and induced significant changes in various aquaporins (AQP1, AQP2, AQP4, and AQP9). In conclusion, betaine supplementation to chicks held for 48 h post-hatch resulted in some changes in blood biochemical indices with no effects on performance during the first 15 days of life. The results suggest that betaine supplementation could ameliorate the stressful effects of water deprivation on POMC and GR expression and maintain cellular osmosis through interactions with variable aquaporins expression, particularly the AQP1 and AQP2. Further investigations are required to investigate the molecular mechanisms underlying the selective regulatory expression of different aquaporins in relation to betaine supplementation.
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The Arabian camel is the largest known mammal that can survive in severe hot climatic conditions. We provide the molecular explanation for the thermotolerance of camel granulosa somatic cells after exposure to 45 °C for 2 (acute heat shock) or 20 h (chronic heat shock). The common features of the cellular responses to acute heat stress were the increase of heat shock proteins and DNA repair enzymes expression. Actin polymerization and Rho signaling were critically activated as a cellular defense against heat shock. Cells exposed to chronic heat shock showed altered cell architecture with a decrease in total detected proteins, metabolic enzymes, and cytoskeletal protein expression. Treatment with transforming growth factor beta (TGFß) pathway inhibitor SB-431542 suppressed the morphological alterations of cells exposed to chronic heat shock. Moreover, during the recovery stage at 38 °C for 24 h, proteomic changes were partially restored with an exponential increase in HSP70 expression, and the cells restored their normal cellular morphology on the 9th day of recovery. Full proteomics data are available via ProteomeXchange with identifier PXD012159. The strategies of cellular defense and tolerance to both thermal conditions reflect the flexible adaptability of camel somatic cells to conserve life under extremely hot conditions.
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The camel is the main meat- and milk-producing animal in the desert environment and is characterized by an induced ovulation pattern, in which ovulation occurs in response to copulation. Little is known about the early embryonic development and placentation in camelid species. Here we describe protocols for the culture of both in vitro-produced and in vivo-retrieved camel embryos. A chemically defined medium enables the development of in vitro-produced embryos from cleavage to the hatching blastocyst stage. In vivo-retrieved embryos will survive in vitro for 23 days postinsemination, reaching a diameter of ~5 mm.
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Blastocisto/metabolismo , Camelidae/embriologia , Meios de Cultura/química , Técnicas de Cultura Embrionária/métodos , Desenvolvimento Embrionário , Indução da Ovulação , Animais , Blastocisto/citologia , Meios de Cultura/farmacologiaRESUMO
Researchers dealing with heat stress experiments use different cell kinds and use trypsin that has been reported to affect the cellular proteins of cultured cells. Therefore, we compared the effects of acute and chronic exposures to high temperature (45 °C) on camel skin fibroblast and granulosa cells. Primary culture of fibroblasts and granulosa cells tolerated the acute heat shock for 2 h; however, granulosa cells cultured for long duration (20 h) showed thermotolerance when compared with the fibroblasts. Moreover, the effect of cell dispersion method (trypsin and mechanical dissociation) on the thermotolerance of sub-cultured cells was examined. Trypsin altered the morphology of fibroblasts and granulosa cells exposed to 45 °C for 4 h. Moreover, trypsin significantly reduced the fibroblast and granulosa cell migration in the wound healing assay. The current results demonstrate that cell passaging and cell type can affect the thermotolerance of the cells; it also revealed that trypsin could alter the cellular response to the heat shock. We raise the demand for another alternative method for cell dispersion in experiments dealing with cellular responses to the heat shock.
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Fibroblastos/metabolismo , Células da Granulosa/metabolismo , Animais , Camelus , Células Cultivadas , Feminino , Febre/metabolismo , Fibroblastos/química , Células da Granulosa/química , Resposta ao Choque Térmico , Temperatura Alta , TermotolerânciaRESUMO
Ten dromedary mature males were used to study the effects of short artificial lighting and low temperature on the reproductive behavior, testicular size, semen quality and hormone during the non-rutting season and subsequent rutting season. Bulls were allocated into two groups: the first group were subjected to natural daylight and temperature and used as a control. The second group was housed individually in light and temperature controlled rooms in which artificial light (300 lux) was used for 10â¯h/d, and the temperature was 25.28⯱â¯0.21⯰C. The trial was initiated in mid-June and continued for 10 weeks in the non-rutting season. The reproductive parameters of all animals in the control and room groups were evaluated once every two weeks. The reproductive parameters of all animals in the control and room groups were re-evaluated during the rutting season of the same year. A significant (P < 0.05) increase in the morphometry of the testes, scrotum, libido, and reaction time score, as well as serum melatonin and testosterone levels, was observed in the treatment non-rutting season (TNRS) group compared to in the control non-rutting season (CNRS) group. The testicular volume, reaction time score, serum melatonin, and testosterone were significantly (P < 0.05) higher in the treatment rutting season (TRS) group than in the control non-rutting season (CRS) group. Improvement in the semen parameters were observed in the TNRS and TRS groups compared to in the CRS group. In conclusion, these results demonstrate that short artificial lighting and low temperature can induce rutting out of season and improve the reproductive parameters of dromedary males during the subsequent rutting season.
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Camelus/fisiologia , Temperatura Baixa , Abrigo para Animais , Iluminação , Reprodução/efeitos da radiação , Animais , Cruzamento/métodos , Camelus/anatomia & histologia , Masculino , Melatonina/metabolismo , Fotoperíodo , Reprodução/fisiologia , Escroto/anatomia & histologia , Escroto/efeitos da radiação , Comportamento Sexual Animal/efeitos da radiação , Testículo/anatomia & histologia , Testículo/efeitos da radiaçãoRESUMO
The present study aimed to examine the ameliorative effects of morin and rutin on the reproductive toxicity induced by titanium dioxide nanoparticles (TiO2NPs) in male rats. A total of seventy adult male Sprague-Dawley rats were randomly divided into seven groups, each comprising ten rats. Nanoreprotoxicity was induced by treating rats with TiO2NPs at a dosage of 300 mg/kg body weight for 30 days. Morin (30 mg/kg body weight) and rutin (100 mg/kg body weight) were co-administered with or without TiO2NPs to rats either individually or combined. Only distilled water was administered to the control group. The results showed that TiO2NPs enhanced oxidative stress, indicated by reduced levels of antioxidants such as superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) in testicular tissues, and increased levels of the lipid peroxidation marker malondialdehyde (MDA). TiO2NPs significantly reduced the levels of sex hormones (testosterone, FSH, and LH), reduced sperm motility, viability, and sperm cell count, and increased sperm abnormalities, in addition to damaging the testicular histological architecture. TiO2NPs resulted in the downregulation of 17ß-HSD and the upregulation of proapoptotic gene (Bax) transcripts in the testicular tissues. Conversely, morin and/or rutin had a protective effect on testicular tissue. They effectively counteracted TiO2NP-induced oxidative damage and morphological injury in the testis by conserving the endogenous antioxidant mechanisms and scavenging free radicals. Thus, we suggest that morin and rutin could be used to alleviate the toxicity and oxidative damage associated with TiO2NP intake.
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Antioxidantes/farmacologia , Flavonoides/farmacologia , Nanopartículas/toxicidade , Rutina/farmacologia , Titânio/toxicidade , Animais , Antioxidantes/metabolismo , Catalase/metabolismo , Glutationa/metabolismo , Glutationa Peroxidase/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Malondialdeído/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Contagem de Espermatozoides , Motilidade dos Espermatozoides/efeitos dos fármacos , Superóxido Dismutase/metabolismo , Testículo/efeitos dos fármacos , Testosterona/metabolismoRESUMO
This study was conducted to evaluate an adjuvant, Montanide (IMS 3015), in improving the quality of Rift Valley fever (RVF) vaccine relative to the traditional adjuvant, aluminum hydroxide gel. Vaccinated sheep were evaluated using biochemical analysis, kidney function tests, liver function tests, and immunological tests. Sheep vaccinated with Montanide (IMS 3015) adjuvant showed significantly higher total protein, total globulin, and gamma globulin concentrations from the second week until the fifth month than the controls. Conversely, albumin concentration and the A/G ratio significantly decreased during this period. Kidney function and liver function tests revealed no differences among any of the groups. There was a significant increase in lymphocyte proportion and a decrease in neutrophil proportion in sheep vaccinated with the Montanide (IMS 3015) adjuvant. Lymphocyte cell proliferation was significantly different in sheep vaccinated with the Montanide (IMS 3015) adjuvant from that in controls. Neutralizing indices were significantly higher in sheep vaccinated with the Montanide (IMS 3015) adjuvant than in controls. The current study showed that sheep vaccinated with inactivated RVF virus with Montanide (IMS 3015) as an adjuvant were protected and no pathological symptoms or biochemical changes were detected. Moreover, the vaccine induced rapid onset of immunological responses with long durations unlike inactivated RVF vaccine with aluminum hydroxide gel.
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Adjuvantes Imunológicos , Óleo Mineral , Vírus da Febre do Vale do Rift/imunologia , Vacinas de Produtos Inativados/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais , Proliferação de Células , Rim/fisiologia , Fígado/fisiologia , Contagem de Linfócitos , Linfócitos/fisiologia , Camundongos , Neutrófilos , Albumina Sérica/metabolismo , Ovinos , Vacinas Virais/efeitos adversos , gama-Globulinas/metabolismoRESUMO
Elite camels often suffer from massive injuries. Thus, there is a pivotal need for a cheap and readily available regenerative medicine source. We isolated novel stem-like cells from camel skin and investigated their multipotency and resistance against various stresses. Skin samples were isolated from ears of five camels. Fibroblasts, keratinocytes, and spheroid progenitors were extracted. After separation of different cell lines by trypsinization, all cell lines were exposed to heat shock. Then, fibroblasts and dermal cyst-forming cells were examined under cryopreservation. Dermal cyst-forming cells were evaluated for resistance against osmotic pressure. The results revealed that resistance periods against trypsin were 1.5, 4, and 7 min for fibroblasts, keratinocytes, and spheroid progenitors, respectively. Furthermore, complete recovery of different cell lines after heat shock along with the differentiation of spheroid progenitors into neurons was observed. Fibroblasts and spheroid progenitors retained cell proliferation after cryopreservation. Dermal cyst-forming cells regained their normal structure after collapsing by osmotic pressure. The spheroid progenitors incubated in the adipogenic, osteogenic, and neurogenic media differentiated into adipocyte-, osteoblast-, and neuron-like cells, respectively. To the best of our knowledge, we isolated different unique cellular types and stem-like cells from the camel skin and examined their multipotency for the first time.
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OBJECTIVES: Endosulfan is a broad-spectrum organochlorine insecticide that has been commercially in use for decades to control insect pests and has been found to pollute the aquatic environment. The current study was carried out to investigate the toxic effects of endosulfan, an organochlorine pesticide, on Nile tilapia (Oreochromis niloticus), a freshwater fish, and the alleviating effects of lycopene on the induced toxicity. METHODS: Four treatment groups of fish were investigated (3 replicates of 15 fish for each group): (1) a control group, (2) a group exposed to endosulfan, (3) a group that was fed on a basal diet supplemented with lycopene, and (4) a group that was fed on a basal diet supplemented with lycopene and exposed to endosulfan. The experiment was carried out over a 4-week period. RESULTS: Endosulfan negatively affected liver function, including liver enzymes and plasma proteins. Endosulfan affected blood parameters of fish and reduced the counts of red blood cells (RBCs) and white blood cells (WBCs), as well as affected immunological parameters. Endosulfan caused oxidative stress, as it decreased the values of antioxidants catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPX), and glutathione (GSH), and increased the level of lipid peroxide malondialdehyde (MDA). Additionally, endosulfan increased cytochrome P450 (CYP450) levels, while it decreased glutathione S-transferase (GST) mRNA transcript levels and distorted the normal histological structure of the liver, gills, and spleen of affected fish. Conversely, lycopene partially restored the aforementioned parameters when administered concomitantly with endosulfan. CONCLUSION: The results showed the beneficial effects of supplementing fish diets with lycopene as a natural antioxidant for ameliorating the toxicity caused by endosulfan.