Insulin-like growth factor-1 improves in vitro meiotic resumption of dromedary camel (Camelus Dromedarius) oocytes

Despite relatively high maturation rate of in vitro matured oocytes in the dromedary camel, however, blastocyst production is very low after in vitro fertilization (IVF). Herein, the influences of oocyte collection method (follicular aspiration vs slicing; Experiment I), the addition of Insulin-like growth factor I (IGF-I) to the maturation medium (Experiment II) on in vitro maturation (IVM) of oocyte were investigated. Although the nuclear maturation did not differ regardless of collecting method, follicular aspiration led to lower degeneration rates than those in controls (P < 0.05). The percentages of oocytes at MII were greater in the presence of IGF-1 than in its absence (71.9% vs 48.4%, respectively, P<0.05). Additionally, the percentages of degenerated oocytes were higher in the control group compared to oocytes cultured in the presence of IGF-I (23.6% vs 10.4%, respectively, P<0.05). IGF-I treatment improved the quality of MII matured oocytes as evidenced by the decrease of cathepsin B (CTSB) activity, a marker of poor quality oocytes, when compared to control ones (P < 0.05). In conclusion, follicular aspiration decreased the degeneration rate; however, it had no effect on completion of maturation. IGF-I enhanced the IVM of oocyte and decreased degeneration rate.(AU)

Amelioration of exosome and mesenchymal stem cells in rats infected with diabetic nephropathy by attenuating early markers and aquaporin-1 expression / Melhoria das células-tronco exossomais e mesenquimais em ratos infectados com nefropatia diabética atenuando marcadores precoces e expressão de aquaporina-1

Diabetic nephropathy (DN) is a prevalent diabetic microvascular condition. It is the leading cause of kidney disease in the advanced stages. There is no currently effective treatment available. This research aimed to investigate the curative potentials of exosomes isolated from mesenchymal stem cells affecting DN. This study was performed on 70 male adult albino rats. Adult rats were randomized into seven groups: Group I: Negative control group, Group II: DN group, Group III: Balanites treated group, Group IV: MSCs treated group, Group V: Exosome treated group, Group VI: Balanites + MSCs treated group and Group VII: Balanites + exosome treated group. Following the trial period, blood and renal tissues were subjected to biochemical, gene expression analyses, and histopathological examinations. Results showed that MDA was substantially increased, whereas TAC was significantly decreased in the kidney in the DN group compared to normal health rats. Undesired elevated values of MDA levels and a decrease in TAC were substantially ameliorated in groups co-administered Balanites aegyptiacae with MSCs or exosomes compared to the DN group. A substantial elevation in TNF-α and substantially diminished concentration of IGF-1 were noticed in DN rats compared to normal health rats. Compared to the DN group, the co-administration of Balanites aegyptiacae with MSCs or exosomes substantially improved the undesirable elevated values of TNF-α and IGF-1. Furthermore, in the DN group, the mRNA expression of Vanin-1, Nephrin, and collagen IV was significantly higher than in normal healthy rats. Compared with DN rats, Vanin-1, Nephrin, and collagen IV Upregulation were substantially reduced in groups co-administered Balanites aegyptiacae with MSCs or exosomes. In DN rats, AQP1 expression was significantly lower than in normal healthy rats. Furthermore, the groups co-administered Balanites aegyptiacae with MSCs or exosomes demonstrated a substantial increase in AQP1 mRNA expression compared to DN rats.

A nefropatia diabética (ND) é uma condição microvascular diabética prevalente. É a principal causa de doença renal em estágios avançados. Atualmente, não há tratamento eficaz disponível. Esta pesquisa teve como objetivo investigar os potenciais curativos de exossomos isolados de células-tronco mesenquimais que afetam a ND. Este estudo foi realizado em 70 ratos albinos adultos machos. Ratos adultos foram randomizados em sete grupos: Grupo I: Grupo de controle negativo, Grupo II: Grupo DN, Grupo III: Grupo tratado com Balanites, Grupo IV: Grupo tratado com MSCs, Grupo V: Grupo tratado com exossomos, Grupo VI: Grupo tratado com Balanites + MSCs e Grupo VII: Balanites + grupo tratado com exossomas. Após o período experimental, o sangue e os tecidos renais foram submetidos a análises bioquímicas, de expressão gênica e exames histopatológicos. Os resultados mostraram que o MDA aumentou substancialmente, enquanto o TAC diminuiu significativamente no rim no grupo DN em comparação com ratos saudáveis normais. Valores elevados indesejados de níveis de MDA e uma diminuição no TAC foram substancialmente melhorados em grupos coadministrados Balanites aegyptiacae com MSCs ou exossomas em comparação com o grupo DN. Uma elevação substancial em TNF-α e uma concentração substancialmente diminuída de IGF-1 foram observadas em ratos DN em comparação com ratos saudáveis normais. Em comparação com o grupo DN, a coadministração de Balanites aegyptiacae com MSCs ou exossomas melhorou substancialmente os valores elevados indesejáveis de TNF-α e IGF-1. Além disso, no grupo DN a expressão de mRNA de vanina-1, nefrina e colágeno IV foi significativamente maior do que em ratos saudáveis normais. Comparado com ratos DN, Vanin-1, Nephrin e colágeno IV Upregulation foram substancialmente reduzidos em grupos co-administrados Balanites aegyptiacae com MSCs ou exossomos. Em ratos DN, a expressão de AQP1 foi significativamente menor do que em ratos saudáveis normais. Além disso, os grupos que coadministraram Balanites aegyptiacae com MSCs ou exossomos demonstraram um aumento substancial na expressão de mRNA de AQP1 em comparação com ratos DN.

Association between somatotropic axis gene polymorphisms and reproductive efficiency of bovine females: a review / Associação entre polimorfismos de genes do eixo somatotrópico e eficiência reprodutiva de fêmeas bovinas: revisão de literatura

Reproductive characteristics directly reflect on productive performance, as they determine the number of calves born annually and the interval between generations, thereby affecting the selection intensity. This allows for the determination of the most important economic variables that directly affect the production system. The present study is a literature review of somatotropic axis gene polymorphisms that are associated with reproductive functions in heifers and cows (Bos taurus taurus and Bos taurus indicus). The following characteristics were analyzed from the various studies reviewed: number of services per pregnancy (Nº services/ pregnancy), age at first calving (AFC), calving interval (CI), and period of service calving conception interval (CCI). The research was conducted during the period from 2011 to 2021, and associations were found among all indicators of reproductive efficiency evaluated in heifers and cows, with 27 single nucleotide polymorphisms (SNPs): 18 in GHR, 5 in IGF-I, and 4 in STAT5A. This study revealed the relationships between SNPs and the reproductive efficiency indicators and demonstrated that molecular genetic techniques enhance the animal selection process, resulting in more profitable systems that are sustainable in the long term.

Características reprodutivas refletem diretamente no desempenho produtivo visto que são capazes de determinar o número de bezerros nascidos anualmente, além de afetar o intervalo entre gerações e, consequentemente, a intensidade de seleção, fazendo com o que tais atributos sejam apontados como variáveis econômicas das mais importantes que afetam diretamente o sistema de produção. O presente estudo propõe realizar uma revisão de literatura de polimorfismos de genes do eixo somatotrópico associados a funções reprodutivas em novilhas e vacas (Bos taurus taurus e Bos taurus indicus), indicadas pelos índices: número de serviços por gestação (Nº serviços/gestação), idade ao primeiro parto (IPP), intervalo de partos (IP) e período de serviço, como também denominado de intervalo parto-concepção (IPC). As pesquisas encontradas durante o período de 2011 a 2021 demonstraram associações entre todos os indicadores de eficiência reprodutiva analisados de novilhas e vacas com 27 polimorfismos de nucleotídeo único (SNPs), sendo 18 em GHR, 5 em IGF-I e 4 em STAT5A. Dessa forma, este estudo mostrou relação entre os SNPs e os indicadores de eficiência reprodutiva, demonstrando que técnicas de genética molecular potencializam o processo de seleção dos animais, resultando em sistemas mais lucrativos e que se sustentam a longo prazo.

Altrenogest during early pregnancy modulates uterine glandular epithelium and endometrial growth factor expression at the time implantation in pigs

Abstract This study evaluated the effects of supplying altrenogest from day 6-12 of pregnancy on the endometrial glandular epithelium, corpora lutea (CL) morphology, and endometrial and CL gene expression. A total of 12 crossbred females (Landrace × Large White) were used. The females were assigned to 4 treatments according to a random design with a 2 × 2 factorial arrangement, with two categories (sow or gilt) and two treatments (non-treated and treated with altrenogest). On day 6 of pregnancy, animals were allocated to one of the following groups: non-treated (NT, n = 6; 3 sows and 3 gilts), and (T, n = 6; 3 sows and 3 gilts) treated daily with 20 mg of altrenogest, from day 6-12 of pregnancy. All animals were euthanized on day 13 of pregnancy. All CLs were individually weighed, and their volume were determined. The endometrial glandular density (GD), mean glandular area (MGA), and vascular density (VD) were determined by histomorphometric and immunohistochemical analyses. Endometrium samples were collected and analyzed by qRT-PCR to evaluate the abundance of transcripts for VEGF and IGF-I. Females in the T group had higher MGA (P < 0.05) compared to the NT group. There was no effect of treatment on GD or VD for both experimental groups. Sows in the T group had augmented expression of IGF-I (P < 0.05). Progestagen had no detrimental effect on CL morphology. In conclusion, altrenogest improves the uterine environment during the peri-implantation period in pigs without compromising corpora lutea development.

Altrenogest during early pregnancy modulates uterine glandular epithelium and endometrial growth factor expression at the time implantation in pigs

This study evaluated the effects of supplying altrenogest from day 6-12 of pregnancy on the endometrial glandular epithelium, corpora lutea (CL) morphology, and endometrial and CL gene expression. A total of 12 crossbred females (Landrace × Large White) were used. The females were assigned to 4 treatments according to a random design with a 2 × 2 factorial arrangement, with two categories (sow or gilt) and two treatments (non-treated and treated with altrenogest). On day 6 of pregnancy, animals were allocated to one of the following groups: non-treated (NT, n = 6; 3 sows and 3 gilts), and (T, n = 6; 3 sows and 3 gilts) treated daily with 20 mg of altrenogest, from day 6-12 of pregnancy. All animals were euthanized on day 13 of pregnancy. All CLs were individually weighed, and their volume were determined. The endometrial glandular density (GD), mean glandular area (MGA), and vascular density (VD) were determined by histomorphometric and immunohistochemical analyses. Endometrium samples were collected and analyzed by qRT-PCR to evaluate the abundance of transcripts for VEGF and IGF-I. Females in the T group had higher MGA (P < 0.05) compared to the NT group. There was no effect of treatment on GD or VD for both experimental groups. Sows in the T group had augmented expression of IGF-I (P < 0.05). Progestagen had no detrimental effect on CL morphology. In conclusion, altrenogest improves the uterine environment during the peri-implantation period in pigs without compromising corpora lutea development.(AU)

Glutathione and IGF-1 in bovine seminal cryopreservation: oxidative stress response and pregnancy rate / [Glutationa e IGF-1 na criopreservação seminal bovina: resposta do estresse oxidativo e taxa de gestação]

The objective of this study was to evaluate the rate of conception, metabolic, and structural conditions of cryopreserved bovine sperm cells, plus extender with IGF-1 and glutathione (GSH). 12 ejaculations of Nelore bulls were used, submitted to treatments: control, gSH (2mM/mL), IGF-1 (100ng/mL) and gSH (1mM/mL) + IGF-1 (50ng/mL). After cryopreservation and thawing the semen passed the fast thermo resistance test (TTR), plasma membrane and acrosomal integrity (PIAI), mitochondrial membrane potential (AP), oxidative stress, and conception rate. Tukey test was used for the statistical analysis of the parametric variables and the Friedman test for nonparametric. The gestation percentage was compared by the Chi-square test. There was no statistical difference (P<0.05) between treatments for the TTRr variable. Otherwise in the oxidative stress evaluated with the CellROX probe was noted that the IGF-1 showed the highest number of reactive cells (P<0.05). The PIAI, AP and gestation rate showed no difference among treatments (P>0.05), with an average of conceptions of 36.58%. It is concluded that IGF-1, gSH and their association did not cause changes in sperm motility, mitochondrial potential, plasma and acrosomal membrane integrity. IGF-1 increased oxidative stress, however, there was no difference in the gestation rate among the treatments.(AU)

Objetivou-se avaliar a taxa de concepção, as condições metabólicas e estrutural das células espermáticas bovinas criopreservadas, acrescidas de diluidores com fator de crescimento semelhante à insulina do tipo 1(IGF-1) e glutationa (GSH). Foram utilizados 12 ejaculados de touros da raça Nelore, submetidos aos tratamentos: controle, gSH (2mM/mL), IGF-1 (100ng/mL) e gSH (1mM/mL) + IGF-1 (50ng/mL). Após a criopreservação e descongelação, o sêmen passou pelos testes de termorresistência rápida (TTRr), integridade de membrana plasmática e acrossomal (PIAI), alto potencial mitocondrial (AP), estresse oxidativo e taxa de concepção. Utilizou-se o teste de Tukey para as análises estatísticas das variáveis paramétricas e o teste de Friedman para as não paramétricas, com significância de 5%. A percentagem de gestação foi comparada pelo teste do qui-quadrado. Não hove diferença estatística (P<0,05) entre os tratamentos para a variável TTRr. Já no estresse oxidativo avaliado com a sonda CellROX, observou-se que o IGF-1 apresentou maior quantidade de células reativas (P<0,05), 36,38± 24,10. A PIAI, o AP e a taxa de gestação não apresentaram diferença entre tratamentos (P>0,05), com média de concepções de 36,58%. Conclui-se que o IGF-1, a gSH e a sua associação não causaram mudanças na motilidade espermática, no potencial mitocondrial, na integridade da membrana plasmática e acrossomal. O IGF-1 aumentou o estresse oxidativo, porém sem diferença na taxa de gestação entre os tratamentos.(AU)

Probiotic supplementation affects IGF-1 and leptin levels in Nile tilapia hepatopancreatic tissue / Suplementação com probiótico afeta os níveis de IGF-1 e leptina no tecido hepatopancreático de tilápias-do-nilo

This work aimed to assess the effect of the probiotic strain, Lactobacillus plantarum, on the levels of leptin, IGF-1 and their receptors on the hepatopancreatic tissues of Nile tilapia (Oreochromis niloticus) and then correlate fish growth performance and gut microbiological parameters. Fish juveniles (±23g) were reared in a recirculation system with constant aeration and temperature (25°C). They were distributed into six polyethylene tanks (45L) and fed twice a day at 5% of the tank biomass with the respective diets: control (commercial diet without probiotic) and supplemented with L. plantarum inoculum (1 x 108 CFU mL-1), both in triplicate. After 30 days of feeding, L. plantarum-fed fishes showed greater weekly growth rate, final weight, and feed conversion rate, in addition to higher count of lactic-acid bacteria and lower count of pathogenic bacteria in the intestinal tract, when compared to the control group. The immunostaining intensity for IGF-1 and leptin hormones was lower after L. plantarum supplementation than in the control group, with no change in the level for receptors. This reduction could implicate important changes in fish metabolism and homeostasis.(AU)

O presente trabalho teve como objetivo avaliar o efeito da cepa probiótica Lactobacillus plantarum sobre os níveis de leptina, IGF-1 e seus receptores no tecido hepatopancreático de tilápia-do-nilo (Oreochromis niloticus) e correlacionar com o desempenho zootécnico e os parâmetros microbiológicos intestinais dos peixes. Juvenis de tilápia-do-nilo (±23g) foram distribuídos em seis tanques de polietileno (45L) conectados a um sistema de recirculação, com aeração e temperatura constantes (25°C). Os peixes foram alimentados duas vezes ao dia, a 5% da biomassa do tanque, com as respectivas dietas: controle (dieta comercial sem probiótico) e suplementada com L. plantarum (1 x 108 UFC mL-1), ambas em triplicata. Após 30 dias de cultivo, os peixes alimentados com L. plantarum apresentaram maiores ganho de peso semanal, peso final e conversão alimentar, bem como maior contagem de bactérias ácido-láticas e menor contagem de bactérias patogênicas no trato intestinal das tilápias alimentadas com dieta probiótica, em comparação ao grupo controle. A intensidade da imunomarcação para os hormônios IGF-1 e leptina foi menor com a suplementação de L. plantarum do que no grupo controle, sem alterar os níveis de seus receptores. Essa redução pode implicar mudanças importantes no metabolismo e na homeostase dos peixes.(AU)

Effect of Supplementary Methionine on Feather Growth and Related Indicators of Pigeon Squabs

We assessed the effect of health sand dietary supplementation with methionine (Met) on White King pigeons. Paired pigeons (n = 180) were fed one of five diets; group T1 received no added Met, while T2, T3, T4 and T5 received 30, 60, 90 and 120 g of supplemental DL-Met/kg, respectively. Each treatment was replicated three times with 24 pairs in each replicate. The results showed that supplementary Met had a minor effect on the length of the fourth primary wing feather in 28-day-old squabs (p>0.05), but the length of 14-day-old squabs in T2 was significantly longer (p=0.010). Dietary Met had a minor effect on Wnt-7a and fibroblast growth factor receptors-2 (FGFR-2) mRNA levels in 28-day-old squabs (p>0.05). The IGF-1 concentration in plasma was highest in T4 and lowest in T2 (p=0.012), but there was no difference between T1, T2 and T5 (p>0.05). In the chest muscle, the expression of IGF-1 in T3 and T4 was higher than in T1 (p=0.172 and 0.015, respectively). In the leg muscle, IGF-1 mRNA level was higher in T4 and T3, and lower in T2 (p>0.05). The results indicate that the optimal Met supplement for increasing fourth primary wing feather length was 30 g/kg Met in health sand, and the feathers were the longest in 14-day-old squabs. Adding 90 g/kg Met to health sand can improve the concentration of IGF-1, which is important for growth performance of pigeon squabs.(AU)

Effect of Supplementary Methionine on Feather Growth and Related Indicators of Pigeon Squabs

We assessed the effect of health sand dietary supplementation with methionine (Met) on White King pigeons. Paired pigeons (n = 180) were fed one of five diets; group T1 received no added Met, while T2, T3, T4 and T5 received 30, 60, 90 and 120 g of supplemental DL-Met/kg, respectively. Each treatment was replicated three times with 24 pairs in each replicate. The results showed that supplementary Met had a minor effect on the length of the fourth primary wing feather in 28-day-old squabs (p>0.05), but the length of 14-day-old squabs in T2 was significantly longer (p=0.010). Dietary Met had a minor effect on Wnt-7a and fibroblast growth factor receptors-2 (FGFR-2) mRNA levels in 28-day-old squabs (p>0.05). The IGF-1 concentration in plasma was highest in T4 and lowest in T2 (p=0.012), but there was no difference between T1, T2 and T5 (p>0.05). In the chest muscle, the expression of IGF-1 in T3 and T4 was higher than in T1 (p=0.172 and 0.015, respectively). In the leg muscle, IGF-1 mRNA level was higher in T4 and T3, and lower in T2 (p>0.05). The results indicate that the optimal Met supplement for increasing fourth primary wing feather length was 30 g/kg Met in health sand, and the feathers were the longest in 14-day-old squabs. Adding 90 g/kg Met to health sand can improve the concentration of IGF-1, which is important for growth performance of pigeon squabs.

Insulina e IGF-1 no meio extensor de criopreservação seminal bovina / Insulin and IGF-I in extender medium for bovine semen cryopreservation

Objetivou-se avaliar a condição metabólica e estrutural das células espermáticas bovinas após congelação, com adição prévia de IGF-I e insulina no meio diluidor seminal. Os ejaculados de seis touros Nelore foram submetidos a quatro tratamentos: controle; insulina (100µUI/mL); IGF-I (150ng/mL) e insulina + IGF-I (50µUI/mL e 75ng/mL, respectivamente). Após a congelação, realizaram-se os testes de termorresistência rápida, coloração pelo corante azul de tripan e Giemsa, além da análise computadorizada da motilidade espermática, da integridade das membranas plasmática e acrossomal, e da peça intermediária por meio de sondas fluorescentes. O teste de termorresistência rápida apresentou efeito dentro do tempo de cada tratamento, mas não entre os tratamentos. Na análise computadorizada da motilidade espermática, foram observados movimento, motilidade e velocidade espermáticos; não houve efeitos dos tratamentos sobre qualquer uma dessas variáveis. Respostas iguais foram obtidas com as sondas fluorescentes e o corante azul de tripan/Giemsa. A adição de insulina e IGF-I, de forma isolada ou combinada, ao meio diluidor para congelação de sêmen não produziu efeitos sobre as condições metabólica e estrutural das células espermáticas.(AU)

This study aimed to evaluate the metabolic and structural condition of the spermatic bovine cells after the freezing, with addition, previously, of IGF-I and Insulin in the seminal thinner medium. The semen of 6 Nellore bulls were submitted to four treatments: Control, Insulin (100µUI/mL); IGF-I (150ng/mL) and Insulin + IGF-I (50µUI/mL and 75ng/mL, respectively). After freezing, rapid resistance tests, Tripan and Giemsa Blue staining, and computerized analysis of sperm motility and integrity of the plasma and acrosomal membranes and the intermediate part were performed by fluorescent probes. The term rapid resistance test had effect within the time of each treatment, but not between treatments. In the computer analysis of sperm motility, sperm movement, motility and velocity no effects of treatments were observed on any of these variables. The same results were obtained with the fluorescent probes and the Blue dye Trypan / Giemsa. The addition of Insulin and IGF-I, alone or in combination, to the semen freezing dilution medium had no effect on the metabolic and structural condition of sperm cells.(AU)

Insulina e IGF-1 no meio extensor de criopreservação seminal bovina / Insulin and IGF-I in extender medium for bovine semen cryopreservation

Objetivou-se avaliar a condição metabólica e estrutural das células espermáticas bovinas após congelação, com adição prévia de IGF-I e insulina no meio diluidor seminal. Os ejaculados de seis touros Nelore foram submetidos a quatro tratamentos: controle; insulina (100µUI/mL); IGF-I (150ng/mL) e insulina + IGF-I (50µUI/mL e 75ng/mL, respectivamente). Após a congelação, realizaram-se os testes de termorresistência rápida, coloração pelo corante azul de tripan e Giemsa, além da análise computadorizada da motilidade espermática, da integridade das membranas plasmática e acrossomal, e da peça intermediária por meio de sondas fluorescentes. O teste de termorresistência rápida apresentou efeito dentro do tempo de cada tratamento, mas não entre os tratamentos. Na análise computadorizada da motilidade espermática, foram observados movimento, motilidade e velocidade espermáticos; não houve efeitos dos tratamentos sobre qualquer uma dessas variáveis. Respostas iguais foram obtidas com as sondas fluorescentes e o corante azul de tripan/Giemsa. A adição de insulina e IGF-I, de forma isolada ou combinada, ao meio diluidor para congelação de sêmen não produziu efeitos sobre as condições metabólica e estrutural das células espermáticas.(AU)

This study aimed to evaluate the metabolic and structural condition of the spermatic bovine cells after the freezing, with addition, previously, of IGF-I and Insulin in the seminal thinner medium. The semen of 6 Nellore bulls were submitted to four treatments: Control, Insulin (100µUI/mL); IGF-I (150ng/mL) and Insulin + IGF-I (50µUI/mL and 75ng/mL, respectively). After freezing, rapid resistance tests, Tripan and Giemsa Blue staining, and computerized analysis of sperm motility and integrity of the plasma and acrosomal membranes and the intermediate part were performed by fluorescent probes. The term rapid resistance test had effect within the time of each treatment, but not between treatments. In the computer analysis of sperm motility, sperm movement, motility and velocity no effects of treatments were observed on any of these variables. The same results were obtained with the fluorescent probes and the Blue dye Trypan / Giemsa. The addition of Insulin and IGF-I, alone or in combination, to the semen freezing dilution medium had no effect on the metabolic and structural condition of sperm cells.(AU)

The interaction of Wnt signaling members with growth factors in cultured granulosa cells

Wnt family members have recently been distinguished in the adult ovary with potential roles in ovarian function. Though particular growth factors interact with Wnt signaling members in extraovarian cell types, it is unclear whether this interaction is applicable in the granulosa cells. Therefore, the current study aimed to determine the effect of insulin-like growth factor-1 (IGF-I), epidermal growth factor (EGF) and basic fibroblast growth factor (FGF-β) on Wnt ligands WNT2 and WNT4 and Wnt receptor Frizzled-4 (FZD4) protein levels in cultured mouse granulosa cells. Granulosa cells were isolated from antral follicles of adult Balb/C mice and cultured for 24 hours in the presence of 100 ng/mL of IGF-I, or EGF or FGF-β. WNT2, WNT4 and FZD4 protein levels were evaluated through western blotting after the culture process. IGF-I treated granulosa cells had significantly the highest level of WNT2 and WNT4 as well as FZD4 when compared to FGF-β and EGF groups. FGF-β group had a significantly higher level of WNT2, WNT4 and FZD4 expression when compared to EGF group. FZD4 expression was at the highest level in the IGF-I group and this difference was statistically significant for all groups including uncultured cells and vehicle group. In addition, FGF-β was shown to positively affect the adhesion of granulosa cells. This study demonstrates that IGF-I, FGF-β and EGF have differential effects on the expressions of WNT2, WNT4, and FZD4 in cultured mouse granulosa cells, suggesting that particular growth factors related to ovarian function might conduct their roles in the ovary through Wnt signaling.

The interaction of Wnt signaling members with growth factors in cultured granulosa cells

Wnt family members have recently been distinguished in the adult ovary with potential roles in ovarian function. Though particular growth factors interact with Wnt signaling members in extraovarian cell types, it is unclear whether this interaction is applicable in the granulosa cells. Therefore, the current study aimed to determine the effect of insulin-like growth factor-1 (IGF-I), epidermal growth factor (EGF) and basic fibroblast growth factor (FGF-β) on Wnt ligands WNT2 and WNT4 and Wnt receptor Frizzled-4 (FZD4) protein levels in cultured mouse granulosa cells. Granulosa cells were isolated from antral follicles of adult Balb/C mice and cultured for 24 hours in the presence of 100 ng/mL of IGF-I, or EGF or FGF-β. WNT2, WNT4 and FZD4 protein levels were evaluated through western blotting after the culture process. IGF-I treated granulosa cells had significantly the highest level of WNT2 and WNT4 as well as FZD4 when compared to FGF-β and EGF groups. FGF-β group had a significantly higher level of WNT2, WNT4 and FZD4 expression when compared to EGF group. FZD4 expression was at the highest level in the IGF-I group and this difference was statistically significant for all groups including uncultured cells and vehicle group. In addition, FGF-β was shown to positively affect the adhesion of granulosa cells. This study demonstrates that IGF-I, FGF-β and EGF have differential effects on the expressions of WNT2, WNT4, and FZD4 in cultured mouse granulosa cells, suggesting that particular growth factors related to ovarian function might conduct their roles in the ovary through Wnt signaling.(AU)

cDNA Cloning of a Bovine Insulin-like growth factor-1 from Egyptian Buffalos and Expression of its Recombinant Protein in Escherichia coli / Clonagem de cDNA de um fator-1 de crescimento semelhante à insulina bovina de búfalos egípcios e expressão de sua proteína recombinante em Escherichia coli

Insulin-like growth factor-1 (IGF-1) is regarded as a crucial clinically significant therapeutic agent against several pathological conditions. Recently, recombinant DNA (rDNA) technology has enabled the production of many drugs of rDNA-origin including IGF-1. Securing a readily available supply of IGF-1 is invaluable to clinical research and biotechnological domains. In this work, the cloning of a full-length bovine IGF-1 cDNA and the successful expression of its cognate recombinant IGF-1 protein is reported. Single-strand cDNA was prepared from liver tissues, through the specific reverse transcription (RT) of IGF-1 mRNA. Subsequently, a PCR amplicon of ~543bp was successfully amplified. Recombinant pTARGET™ vector harboring IGF-1 insert was successfully cloned into competent E. coli JM109 cells. SDS-PAGE analysis revealed that the recombinant IGF-1 has been expressed at the expected size of 7.6kDa. The outcome provides a robust basis for transecting the recombinant pTARGETTM vector, harboring the IGF-1 cDNA insert, into mammalian cells. Optimal initial glucose concentration was found to be 10g/l with corresponding protein concentration of 6.2g/l. The proliferative biological activity crude recombinant IGF-1 protein was verified on HeLa cell lines. This is envisaged to facilitate large-scale production of recombinant IGF-1 protein, thereby enabling thorough investigation of its clinical and pharmaceutical effects.(AU)

O fator de crescimento semelhante à insulina-1 (IGF-1) é considerado um agente terapêutico clinicamente significativo contra várias condições patológicas. Recentemente, a tecnologia de DNA recombinante (rDNA) permitiu a produção de muitos medicamentos de origem rDNA, incluindo o IGF-1. Garantir um suprimento prontamente disponível de IGF-1 é inestimável para pesquisas clínicas e domínios biotecnológicos. Neste trabalho, relata-se a clonagem de um cDNA de IGF-1 bovino de comprimento total e a expressão bem-sucedida de sua proteína IGF-1 recombinante cognata. O cDNA de cadeia simples foi preparado a partir de tecidos do fígado, por meio da transcrição reversa específica (RT) do mRNA de IGF-1. Posteriormente, um amplificador de PCR de ~ 543pb foi amplificado com sucesso. O vetor pTARGET™ recombinante contendo a inserção de IGF-1 foi clonado com sucesso em células competentes E. coli JM109. A análise por SDS-PAGE revelou que o IGF-1 recombinante foi expresso no tamanho esperado de 7,6kDa. O resultado fornece uma base robusta para a transferência do vetor pTARGETTMTM recombinante, abrigando a inserção de cDNA de IGF-1 em células de mamíferos. Verificou-se que a concentração inicial ideal de glicose é 10g/L, com a concentração de proteína correspondente de 6,2g/L. A proteína IGF-1 recombinante bruta de atividade biológica proliferativa foi verificada nas linhas celulares HeLa. É previsto que isso facilite a produção da proteína IGF-1 recombinante em larga escala, permitindo, assim, uma investigação completa dos seus efeitos clínicos e farmacêuticos.(AU)

cDNA Cloning of a Bovine Insulin-like growth factor-1 from Egyptian Buffalos and Expression of its Recombinant Protein in Escherichia coli / Clonagem de cDNA de um fator-1 de crescimento semelhante à insulina bovina de búfalos egípcios e expressão de sua proteína recombinante em Escherichia coli

Insulin-like growth factor-1 (IGF-1) is regarded as a crucial clinically significant therapeutic agent against several pathological conditions. Recently, recombinant DNA (rDNA) technology has enabled the production of many drugs of rDNA-origin including IGF-1. Securing a readily available supply of IGF-1 is invaluable to clinical research and biotechnological domains. In this work, the cloning of a full-length bovine IGF-1 cDNA and the successful expression of its cognate recombinant IGF-1 protein is reported. Single-strand cDNA was prepared from liver tissues, through the specific reverse transcription (RT) of IGF-1 mRNA. Subsequently, a PCR amplicon of ~543bp was successfully amplified. Recombinant pTARGET™ vector harboring IGF-1 insert was successfully cloned into competent E. coli JM109 cells. SDS-PAGE analysis revealed that the recombinant IGF-1 has been expressed at the expected size of 7.6kDa. The outcome provides a robust basis for transecting the recombinant pTARGETTM vector, harboring the IGF-1 cDNA insert, into mammalian cells. Optimal initial glucose concentration was found to be 10g/l with corresponding protein concentration of 6.2g/l. The proliferative biological activity crude recombinant IGF-1 protein was verified on HeLa cell lines. This is envisaged to facilitate large-scale production of recombinant IGF-1 protein, thereby enabling thorough investigation of its clinical and pharmaceutical effects.(AU)

O fator de crescimento semelhante à insulina-1 (IGF-1) é considerado um agente terapêutico clinicamente significativo contra várias condições patológicas. Recentemente, a tecnologia de DNA recombinante (rDNA) permitiu a produção de muitos medicamentos de origem rDNA, incluindo o IGF-1. Garantir um suprimento prontamente disponível de IGF-1 é inestimável para pesquisas clínicas e domínios biotecnológicos. Neste trabalho, relata-se a clonagem de um cDNA de IGF-1 bovino de comprimento total e a expressão bem-sucedida de sua proteína IGF-1 recombinante cognata. O cDNA de cadeia simples foi preparado a partir de tecidos do fígado, por meio da transcrição reversa específica (RT) do mRNA de IGF-1. Posteriormente, um amplificador de PCR de ~ 543pb foi amplificado com sucesso. O vetor pTARGET™ recombinante contendo a inserção de IGF-1 foi clonado com sucesso em células competentes E. coli JM109. A análise por SDS-PAGE revelou que o IGF-1 recombinante foi expresso no tamanho esperado de 7,6kDa. O resultado fornece uma base robusta para a transferência do vetor pTARGETTMTM recombinante, abrigando a inserção de cDNA de IGF-1 em células de mamíferos. Verificou-se que a concentração inicial ideal de glicose é 10g/L, com a concentração de proteína correspondente de 6,2g/L. A proteína IGF-1 recombinante bruta de atividade biológica proliferativa foi verificada nas linhas celulares HeLa. É previsto que isso facilite a produção da proteína IGF-1 recombinante em larga escala, permitindo, assim, uma investigação completa dos seus efeitos clínicos e farmacêuticos.(AU)

Foals developmental orthopedic disease associated with metabolic and biometric characterization of pregnant overweight crioulo mares

Background: Musculoskeletal changes in growing foals can be linked to metabolic disorders which affect the cartilage metabolism associated of obesity during the late gestation of the mares, negatively affecting the athletic performance of the otherwise prospective foals. High basal insulin levels can be associated with increased weight and obesity of the mares, altering the supply of the glucose to the fetus and the production of IGF-1, which plays an important role in the endochondral cartilage metabolism. The present study aims to describe the association of metabolic and biometric alterations in overweight Criollo mares with IGF-1 levels and the presence of articular and physeal lesions in their foals. Materials, Methods & Results: A prospective and observational cohort study was conducted using the foals and the overweight mares at field conditions. Twenty-eight foals and their overweight mothers were utilized. The foals’ biometric and metabolic features as weight and height; glucose, total cholesterol, cholesterol HDL, triglycerides, basal insulin, leptin, glucose, total T4 and IGF-1 were analyzed, from the birth until the weaning period around sixth month of life of it. All the foals, at the weaning period, were submitted to the radiograph examination of the tarsal joints and the metacarpal-phalangeal joints to evaluate signs of lesions and the possible distal metacarpal physeal changes, respectively. The mares’ biometric and metabolic features as weight, height and fat tail-head deposition; basal insulin, leptin, glucose, total cholesterol, cholesterol HDL and triglycerides were analyzed at the final period of gestation. The basal insulin levels of the foals at the second, fourth and sixth month of life were 9.87 ± 1.82 µUI/mL, 9.13 ± 1.94 µUI/mL, 9.39 ± 2.54 µUI/mL. The IGF-1 levels of the foals at the second, third, fourth, fifth and sixth...

Blood parameters of Nellore steers fed a high concentrate diet in two restricted feeding regimes followed by ad libitum feeding / Parâmetros sanguíneos de novilhos Nelore submetidos à dieta de alto concentrado em dois regimes alimentares sequenciais (fase 1: restrição e fase 2: alimento ad libitum)

Blood parameters of 18 Nellore steers submitted to high concentrate diet in two sequential feeding regimes (phase 1: restriction and phase 2: ad libitum feeding) were evaluated. In the first phase, steers were fed for 56 days three intake levels of dry mat-ter (ad libitum, 75 g dry matter (DM)/kg metabolic body weight (BW0.75) or 60 g DM/kg BW(0.75). In the second phase, from the 57th day on feed, animals fed 60 g DM/kg BW(0.75) and 75 g DM/kg BW(0.75) were fed ad libitum until the 136th day. In the 8th, 27th, 55th (first phase), 71st, 84th, 112th and 132nd (second phase) day of experimental period, blood samples were collected and insulin-like growth factor I (IGF-I), triiodothyronine (T3), thyroxine (T4), glucose, plasma ureic nitrogen (PUN) and total protein (TP) were determined. In the first phase IGF-I, T4, glucose and total protein concentration was not different among treat-ments (P>0.05) and there was no interaction between treatment and time (P>0.05). There was an interaction among treatments and time on T3 and plasma ureic nitrogen (P<0.01). In the second phase IGF-I, T3, T4, glucose and total protein concentration was not different among treatments and there was no interaction between treatment and time (P>0.05). For plasma ureic nitrogen concentration there was an interaction among treatments and time (P=0.02). Lower levels of diet intake can be used to improve animal performance in feedlot.(AU)

Foram avaliados parâmetros sanguíneos de 18 novilhos Nelore submetidos a dois regimes sequenciais de alimentação (fase 1: restrição e fase 2: alimento ad libitum) com dieta de alto teor de concentrado. Na primeira fase, os animais foram alimentados por 56 dias em três níveis de ingestão de matéria seca (MS) representando os tratamentos: ad libitum, 75 g MS/kg peso corporal metabólico (PV0,75) e 60 g MS/kg PV(0,75). Na segunda fase, a partir do 57º dia de confinamento, os animais alimentados com 60 g MS/kg PV(0,75) e 75 g MS/kg PV(0,75) receberam alimentação ad libitum até o 136º dia. No 8º, 27º, 55º, primeira fase, e 71º, 84º, 112º e 132º dia de confinamento, segunda fase, foram colhidas amostras de sangue para determinação das concentrações de fator de crescimento semelhante à insulina I (IGF-I), triiodotironina (T3), tiroxina (T4), glicose, nitrogênio ureico plasmático e proteína total. No período de restrição alimentar as concentrações plasmáticas de IGF-I, T4, glicose e proteína total não foram diferentes entre os tratamentos (P>0,05) e não houve interação entre tratamento e tempo (P>0,05). Foi observada interação entre tratamento e tempo para a concentração plasmática de T3 e nitrogênio ureico no plasma (P<0,01). No período de alimento ad libitum as concentrações de IGF-I, T3, T4, glicose e proteína total não foram diferentes entre os tratamentos e não houve interação entre tratamento e tempo (P>0,05). Para a concentração de nitrogênio ureico no plasma houve interação entre os fatores tratamentos e tempo (P=0,02). Níveis mais baixos de ingestão de dieta podem ser usados para melhorar o desempenho animal em confinamento.(AU)

Blood parameters of Nellore steers fed a high concentrate diet in two restricted feeding regimes followed by ad libitum feeding / Parâmetros sanguíneos de novilhos Nelore submetidos à dieta de alto concentrado em dois regimes alimentares sequenciais (fase 1: restrição e fase 2: alimento ad libitum)

Blood parameters of 18 Nellore steers submitted to high concentrate diet in two sequential feeding regimes (phase 1: restriction and phase 2: ad libitum feeding) were evaluated. In the first phase, steers were fed for 56 days three intake levels of dry mat-ter (ad libitum, 75 g dry matter (DM)/kg metabolic body weight (BW0.75) or 60 g DM/kg BW(0.75). In the second phase, from the 57th day on feed, animals fed 60 g DM/kg BW(0.75) and 75 g DM/kg BW(0.75) were fed ad libitum until the 136th day. In the 8th, 27th, 55th (first phase), 71st, 84th, 112th and 132nd (second phase) day of experimental period, blood samples were collected and insulin-like growth factor I (IGF-I), triiodothyronine (T3), thyroxine (T4), glucose, plasma ureic nitrogen (PUN) and total protein (TP) were determined. In the first phase IGF-I, T4, glucose and total protein concentration was not different among treat-ments (P>0.05) and there was no interaction between treatment and time (P>0.05). There was an interaction among treatments and time on T3 and plasma ureic nitrogen (P0.05). For plasma ureic nitrogen concentration there was an interaction among treatments and time (P=0.02). Lower levels of diet intake can be used to improve animal performance in feedlot.

Foram avaliados parâmetros sanguíneos de 18 novilhos Nelore submetidos a dois regimes sequenciais de alimentação (fase 1: restrição e fase 2: alimento ad libitum) com dieta de alto teor de concentrado. Na primeira fase, os animais foram alimentados por 56 dias em três níveis de ingestão de matéria seca (MS) representando os tratamentos: ad libitum, 75 g MS/kg peso corporal metabólico (PV0,75) e 60 g MS/kg PV(0,75). Na segunda fase, a partir do 57º dia de confinamento, os animais alimentados com 60 g MS/kg PV(0,75) e 75 g MS/kg PV(0,75) receberam alimentação ad libitum até o 136º dia. No 8º, 27º, 55º, primeira fase, e 71º, 84º, 112º e 132º dia de confinamento, segunda fase, foram colhidas amostras de sangue para determinação das concentrações de fator de crescimento semelhante à insulina I (IGF-I), triiodotironina (T3), tiroxina (T4), glicose, nitrogênio ureico plasmático e proteína total. No período de restrição alimentar as concentrações plasmáticas de IGF-I, T4, glicose e proteína total não foram diferentes entre os tratamentos (P>0,05) e não houve interação entre tratamento e tempo (P>0,05). Foi observada interação entre tratamento e tempo para a concentração plasmática de T3 e nitrogênio ureico no plasma (P0,05). Para a concentração de nitrogênio ureico no plasma houve interação entre os fatores tratamentos e tempo (P=0,02). Níveis mais baixos de ingestão de dieta podem ser usados para melhorar o desempenho animal em confinamento.

Ramipril can alleviate the accumulation of renal mesangial matrix in rats with diabetic nephropathy by inhibiting insulin-like growth factor-1

Purpose:To investigate the impact of Ramipril (RAM) on the expressions of insulin-like growth factor-1 (IGF-1) and renal mesangial matrix (RMM) in rats with diabetic nephropathy (DN).Methods:The Sprague Dawley rats were divided into normal control (NC) group (n = 12), DN group (n = 11), and DN+RAM group (n = 12). The ratio of renal weight to body weight (RBT), fasting blood glucose (FBG), HbA1c, 24-h urine protein (TPU), blood urea nitrogen (BUN), creatinine (Cr), renal pathological changes, the levels of IGF-1, fibronectin (FN), type IV collagen (Col-IV), and matrix metalloproteinases (MMP)-2 were compared among the groups.Results:Compared with NC group, the RBT, FBG, HbA1c, TPU, BUN, Cr, and RMM in DN group were significantly increased (P < 0.05), the IGF-1, FN, and Col-IV were significantly upregulated (P < 0.05), while MMP was significantly downregulated (P < 0.05). Compared with DN group, the indexes except for the FBG and HbA1c in DN+RAM group were significantly improved (P < 0.05), among which IGF-1 exhibited significant positive correlation with TPU(r=0.937), FN(r=0.896) and Col-IV(r=0.871), while significant negative correlation with MMP-2 (r=-0.826) (P<0.05).Conclusion:RAM may protect the kidneys by suppressing IGF-1 and mitigating the accumulation of RMM.(AU)

Características de carcaça, órgão vomeronasal, concentrações séricas de testosterona e IGF-I em machos Nelore com ductos incisivos bloqueados / Characteristics of carcass, vomeronasal organ, serum testosterone and IGF-I concentrations in Nellore males with blocked incisor ducts

Objetivou-se avaliar características quali - quantitativas da carcaça em machos Nelore, submetidos ao bloqueio dos ductos incisivos no período pré puberal. Além disso, objetivou-se avaliar as concentrações séricas de testosterona e do hormônio do crescimento semelhante a insulina do tipo I (IGF-I), e a arquitetura histológica do órgão vomeronasal (OVN). Trinta e quatro machos, no período pré puberal, foram divididos em três grupos experimentais: inteiros ­ animais controle sem qualquer procedimento (n = 11); bloqueados - animais com os ductos incisivos bilateralmente obstruídos (n = 10); castrados - animais com orquiepididectomia bilateral (n = 13). O OVN foi obtido no abate, ao final do experimento, para avaliação histomorfométrica. As coletas de sangue foram realizadas a cada três meses, em dois turnos (manhã e tarde), totalizando cinco avaliações. Observaram-se maiores (P<0,05) pesos corporais finais (inteiros = 494,1 ± 28,71; bloqueados = 500,6 ± 23,6 e castrados = 468,3 ± 21,8 Kg) nos inteiros e bloqueados. O acabamento da carcaça foi maior nos castrados (P<0,05) em relação aos inteiros e bloqueados, enquanto o rendimento de carcaça não apresentou diferenças entre os três tratamentos (P>0,05). Registrou-se redução da altura do epitélio neuro sensitivo do OVN nos bloqueados e castrados (P<0,05). Foi registrada menor concentração de testosterona nos bloqueados nas coletas quatro e cinco à tarde (P<0,05). Conclui-se que a obstrução dos ductos incisivos reduziu os estímulos sensoriais para o OVN, que por sua vez, promoveu alteração na concentração sérica de testosterona, sem alterar o ganho de peso dos animais, porém, não promoveu melhoria na qualidade da carcaça nas condições deste estudo.

A total of 34 prepuberal Nellore males were divided into three groups: control ­ animals without any procedure (n = 11); blocked - with bilaterally incisors ducts obstructed (n = 10); castrated - with bilateral orchiectomy (n = 13). This study aimed to evaluate the effect of blocking the vomeronasal organ (VNO) in qualitative and quantitative characteristics of the carcass. Also, this study aimed to evaluate testosterone and insulin-like growth factor (IGF-I) serum concentration, and the histological architecture of the OVN. Blood samples were taken every three months during two daily collections (morning and afternoon), totalizing five evaluations. It was observed higher (P<0.05) final body weight (control= 494.1 ± 28.71; blocked = 500.6 ± 23.6; castrated = 468.3 ± 21.8 kg) at the control and blocked groups. Carcass finishing was higher in castrated animals (P <0.05), while carcass yield did not differ between treatments (P> 0.05). It was observed a reduction (P<0.05) of the VNO sensory epithelium height in blocked and castrated groups compared with control group. It was registered lower (P<0.05) serum testosterone concentration in blocked group at the fourth and fifth blood collection (afternoon). It was concluded that the obstruction of the incisive ducts reduced the sensorial stimuli for the OVN, which, in turn, promoted a change in the serum concentration of testosterone, without altering the weight gain of the animals, but did not promote improvement in the quality of the carcass under the conditions of this study.