Effect of sugammadex administration on neural tube development in 48-h chick embryos.

Sugammadex is a new generation drug that has led to significant changes in the practice of anesthesia. However, its effects on fetal development are not yet fully known. The aim of this study is to investigate the teratogenic effects of sugammadex on neural tube and embryonic development in early chick embryos. In this study, 50 0-day fertile specific non-pathogenic (SPF) eggs were used. Fifty eggs were divided into 5 different groups, each consisting of 10 pieces. While no substance was given to the control group at the 28th hour of the study, 4 different doses of sugammadex were administered to the experimental groups, respectively 2, 4, 8, 16 mg/kg. Cranio-caudal lengths of embryos, somite numbers, average number of argyrophilic nucleolar regulatory regions (AgNOR) per nucleus, total AgNOR area/total nuclear area (TAA/NA) ratios, Caspase-3 H-Score results, and presence of neural tube defect were compared among the groups. While the mean cranio-caudal lengths, somite counts, TAA/NA ratios and AgNOR counts of the embryos were found to be statistically significantly lower than the control group, Caspase-3 H-Score mean results were found to be significantly higher (p < .05). In addition, it was observed that there was an increase in neural tube patency and developmental delay. As a result, sugammadex crossing the placenta was revealed to increase the release of proapopitotic molecules and disrupt the developmental stages of embryos. Thus, it was determined that sugammadex in increased developmental delay and incidence of neural tube defects in early chick embryos with increased dose dependent. Despite these results, the effects of sugammadex on fetal development in in vivo and in vitro environments should be studied with further studies. RESEARCH HIGHLIGHTS: Sugammadex is a new generation drug that has led to significant changes in the practice of anesthesia. However, its effects on fetal development are not yet fully known. It has been observed that different doses of sugammadex increase the risk of neural tube defect development on chick embryos and slow the embryo development in a dose-dependent manner.

Protective Effects of Different Doses of Ginsenoside-Rb1 Experimental Cataract Model That in Chick Embryos.

PURPOSE: There has been increased interest in phytochemical antioxidants to prevent protein damage and aggregate formation in cataract treatment. In this study, the protective effect of different doses of Rb1 (GRb1), one of the ginsenosides of Panax Ginseng, in the experimental cataract model formed in chick embryos was investigated. METHODS: Five different experimental groups were formed with 100 SPF fertilized eggs: Control (0.9% NaCl to physiological saline), hydrocortisone hemisuccinate sodium (HC), low dose (HC + L-GRb1 (1 mg/kg)), medium dose (HC+). M-GRb1 (2.5 mg/kg)), and high dose (HC + H-GRb1 (5 mg/kg)). All solutions were given to air sack at 15 days of incubation. On the 17th day, the bulbous oculi of the chick embryos were dissected. Cataract formations of the lenses, glutathione (GSH), malondialdehyde (MDA), total antioxidant (TAS), total oxidant (TOS) levels, Caspase-3 H-score, and TUNEL index were determined. In addition, crystalline alpha A (CRYAA) gene expression was evaluated. RESULTS: Cataracts were observed in the control, HC, HC + L-GRb1, HC + M-GRb1, and HC + H-GRb1 groups with a frequency of 0%, 100%, 75%, 56.25%, and 100%, respectively. There were statistically significant differences between the control and HC groups in terms of TAS, TOS, MDA, GSH, Caspase-3 H-score, and TUNEL index (p < .05). When the therapeutic effect of the GRb1 groups was evaluated, the HC group showed significant differences with the HC + L-GRb1 and HC + M-GRb1 groups in almost all parameters (p < .05), while there was no statistical difference with the HC + H-GRb1 group (p > .05). In addition, gene expression levels differed between the groups, although not statistically significant (p > .05). CONCLUSION: 1 mg/kg and 2.5 mg/kg GRb1 applications show therapeutic properties on the HC-induced cataract model. This effect is more pronounced at 2.5 mg/kg.

Automatic segmentation of the cerebellum using volBrain software in normal paediatric population.

BACKGROUND: Morphological differences that can lead to cerebellar volume changes are associated with the pathogenesis of paediatric diseases. The aim of this study was to examine cerebellum volume in a healthy paediatric population. MATERIALS AND METHODS: To provide MRI-based volumetric measurements of the cerebellum, images from the years 2019 to 2021 were scanned retrospectively. A total of 100 images, including the paediatric population aged 0-15 years, were imported into the volBrain software. Volumetric segmentations were obtained automatically, and each lobular cerebellar volume was obtained. The samples were divided into groups of 0-2 years (n = 18), 3-5 years (n = 24), 6-11 years (n = 34) and 12-15 years (n = 24). Obtained cerebellar volumes, age groups, gender and bilateral side comparisons were made. RESULTS: In the comparative analyses performed for the total cerebellum and each of the 12 lobular segments, statistically significant differences were found between the age groups in all measurements except Crus II, lobules VIIB, VIIIA and VIIIB (p < 0.05). In multiple comparison tests, statistically significant differences were found between defined age groups, especially infants and toddlers and early adolescence groups (p < 0.05). There was a significant positive correlation between the ages of the subjects and their cerebellum volumes (p < 0.05). Statistically significant differences were found in lobules I-II, VI, VIIIB, IX and X in right and left side volumes (p < 0.05). CONCLUSION: There is a tendency to increase in cerebellar volume during the transition from childhood to adolescence. The cerebellum has volumetric differences in the first years of life and during adolescence. When the development of a healthy cerebellum is analysed based on volumetric segmentation, differences are observed. The findings of this study may be useful in confirming various theories attributed to the cerebellum in the clinic.

Use of AgNOR staining to determine the effect of metoclopramide on neural tube development in early chick embryos.

Nausea and vomiting during pregnancy are common problems and prolonged pharmacological treatment often is needed; however, the teratogenic effects of anti-emetic drugs on neural tube (NT) development are not clear. We investigated the effects of different doses of metoclopramide on NT development in 48 and 72 h chick embryos using an argyrophilic nucleolar organizing region (AgNOR) staining method. We used 150 fertile, specific pathogen-free eggs incubated for 28 h, then randomly divided into five equal groups: group A, sham control was administered 0.9% saline; groups B - E were administered 0.15 mg/egg, 0.3 mg/egg, 0.6 mg/egg and 1.2 mg/egg, respectively. Half of the eggs in each group were taken from the incubator at 48 h incubation and the other half at 72 h incubation. After incubation, eggs were opened, embryos were dissected from their membranes, fixed with 10% formalin and examined by light microscopy. The NT status, i.e., open or closed, and somite number, crown-rump length, morphological features and gross developmental abnormalities were recorded. Excised embryos were sectioned and stained using hematoxylin and eosin or the AgNOR procedure and examined for morphology and histopathology. Delayed NT closure was observed in all 48 h drug exposed embryos, but in the 72 h groups, this occurred only in high-dose groups. Somite number was reduced significantly in groups C - E compared to the control group. Crown-rump length was decreased in both 48 and 72 h embryos. We found a decreased total AgNOR area:nuclear area ratio in 48 and 72 h embryos of all experimental groups. We found that metoclopramide delayed NT closure in chick embryos in a dose-dependent manner.

Investigation of the antioxidant effect of Chrysin in an experimental cataract model created in chick embryos.

PURPOSE: Cataract, which occurs as a result of lens opacification, is one of the most common causes of vision loss. In the literature, deterioration of the antioxidant system due to the increase in reactive oxygen species and oxidant levels is shown among the causes of cataract formation. The aim of this study was to investigate the antioxidant effect of chrysin on steroid-induced cataract development in an experimental chick embryo model using morphological, histological and biochemical parameters. METHODS: Within the scope of the study, 150 specific pathogen free (SPF) fertilized eggs were used. Eggs were divided into 6 groups as control (group 1), corn oil (group 2), hydrocortisone hemisuccinate sodium (HC) (group 3), low dose chrysin (group 4), medium dose chrysin (group 5) and high dose chrysin (group 6). On the 15th day of incubation, Chrysin and HC were applicated to the air sac of the eggs with Hamilton and/or insulin injector. On day 17, the chick embryos were removed from the eggs and the bulbus oculi of the embryos were dissected. Lenses of 9 embryos were used for morpholigical cataract grading in each group, lens of 8 embryos for biochemical analysis and intact eyes of 7 embryos for histological evaluation (TUNEL method). RESULTS: No opacity was observed in any of the lenses in Group 1 and 2. Cataract was observed in all lenses in Group 3. The mean opacity grades in group 3 were statistically significantly higher when compared to group 1 and 2 (p<0.05). The difference between group 6 and group 3 was statistically significant (p<0.05). GSH and TAS levels in the lenses were statistically significantly decreased compared to the control group due to HC application (p<0.05). It was determined that the decreased GSH and TAS levels in the lenses increased in relation to the Chrysin application doses. The increased levels of MDA, TOS, caspase 3 and caspase 9 in the HC group decreased significantly depending to the chrysin doses (p<0.05). In addition, while the rate of apoptotic cells determined by the TUNEL method was statistically significantly higher in the HC administered group than in the control group (p<0.05), it was statistically significantly decreased in the chrysin-administered groups, in relation to the dose of chrysin (p<0.05). CONCLUSIONS: We think that anti-cataract effect of crhysin may be due to the antioxidant and antiapoptotic properties of chrysin. However, more research is needed to clarify the anti-cataract effects of chrysin.

Investigation of developmental toxicity of favipiravir on fetal bone and embryonic development.

BACKGROUND: Favipiravir is one of the essential antiviral drugs used for the treatment of coronavirus disease (COVID-19) in some countries. However, there is not enough information about used, especially in pregnancy. Therefore, in this study, it was aimed to determine the developmental toxicity of favipiravir on fetal bone development and embryonic development. METHODS: In this study, 16 pregnant wistar albino rats were used. The rats were divided into four groups: Control (saline) and Group A (50 mg/kg × 5 days), Group B (50 mg/kg × 1 days + 20 mg/kg × 4 days), Group C (20 mg/kg × 5 days). Solutions were administered to the rats by oral gavage from the 10th to 14th days of pregnancy, twice a day. The skeletal system development of fetuses was examined with double skeletal staining and immunohistochemical staining methods. RESULTS: A total of 72 fetuses from pregnant rats, 18 in each group, were included in the study. As a result, depending on favipiravir dose increase, in experimental groups, it was determined that the statistically significant decrease on the ossification rates of anterior and posterior extremity bones, and length and weight of fetuses. CONCLUSION: Exposure to favipiravir during pregnancy impairs bone metabolism and bone formation-resorption stages and may cause developmental delay.

Investigation of teratogenic effects of letrozole on fetal bone development / Investigación de los efectos teratogénicos de letrozol en el desarrollo óseo fetal

SUMMARY: Letrozole is mainly used for the treatment of unexplained infertility, breast cancer and polycystic ovarian syndrome, with secondary use in ovarian stimulation. In cases of unexpected or unknown pregnancy during the use of letrozole, letrozole may cause a teratogenic effect on the fetus. In this reason, in this study, we aimed to determine the effect of letrozole on fetal bone development. In this study, 32 pregnant Wistar albino rats were used. The rats were divided into four groups: Control (saline) and high; 0.3 mg/kg, medium; 0.03 mg/kg, low; 0.003 mg/ kg letrozole. Saline and letrozole were administered in 100 mL solutions by intraperitonaly from day 11 to day 15 of pregnancy. The skeletal system development of fetuses was examined with double skeletal staining, immunohistochemical staining methods and mineral density scanning electron microscopy. A total of 100 fetuses from female rats, 25 in each group, were included in the study. As a result of that, ossification rates were observed to decrease depending on the dose of letrozole in the forelimb limb (scapula, humerus, radius, ulna) and hindlimb (femur, tibia, fibula) limb bones. As a result of the statistical analysis, a statistically significant decrease was found in the ossification rates of all bones between the control group and low, medium, high letrozole groups (p<0.001). Exposure to letrozole during pregnancy adversely affected ossification and bone growth. However, the teratogenic effects of letrozole are unclear. Therefore, it needs to be investigated more extensively.

RESUMEN: Letrozol se usa principalmente para el tratamiento de la infertilidad inexplicable, el cáncer de mama y el síndrome de ovario poliquístico, con estimulación ovárica de uso secundario. En casos de embarazo inesperado o desconocido durante el uso de letrozol, puede causar un efecto teratogénico en el feto. Por esta razón, en este estudio, nuestro objetivo fue determinar el efecto de letrozol en el desarrollo óseo fetal. Se utilizaron 32 ratas albinas Wistar preñadas las cuales se distribuyeron en cuatro grupos: Control (solución salina) y alta; 0,3 mg/kg, medio; 0,03 mg/kg, bajo; 0,003 mg/kg de letrozol. Se administró solución salina y letrozol en soluciones de 100 mL por vía intraperitoneal desde el día 11 hasta el día 15 de la preñez. El desarrollo del sistema esquelético de los fetos se examinó con tinción esquelética doble, métodos de tinción inmunohistoquímica y microscopía electrónica de barrido de densidad mineral. Se incluyeron en el estudio un total de 100 fetos de ratas hembra, 25 en cada grupo. Como resultado, se observó que las tasas de osificación disminuían dependiendo de la dosis de letrozol en los huesos de los miembros torácicos (escápula, húmero, radio, ulna) y de las miembros pélvicos (fémur, tibia, fíbula). Se encontró una disminución estadísticamente significativa en las tasas de osificación de todos los huesos entre el grupo control y los grupos de letrozol bajo, medio y alto (p<0,001). La exposición a letrozol durante la preñez afectó negativamente la osificación y el crecimiento óseo. Sin embargo, los efectos teratogénicos del letrozol no están claros por lo que debe ser investigado más extensamente.

A novel model of early type 1 diabetes mellitus: The chick embryo air sack model.

Diabetes Mellitus (DM) is a metabolic disease characterized by hyperglycemia. Chronic hyperglycemia is associated with long-term dysfunction such as retinopathy, nephropathy, neuropathy and cardiovascular diseases. These complications increase rates of death and disability worldwide. Due to the negative effects of DM on the quality of life, the mechanism and treatments of the disease should be investigated in more detail. Most of the research in diabetes is performed in experimental animals. Experimental animal models contributed to the advancement of clinical research, the development of new therapeutic approaches, the discovery of insulin and the purification of insulin. There are many animal models of DM in the literature. But there are a few DM model studies created with chick embryos. In these studies, it was seen that there were differences in STZ doses and STZ administration techniques. The objective of this study was to create a more acceptable and easier DM model. 180 specific pathogen free (SPF) fertilized chicken eggs (White Leghorn chicken) were used in this study. STZ was administered to 160 SPF eggs for an induced DM model. The remaining 20 SPF eggs were separated as a control group. We used two different DM models (Air sack model (ASM) and Chorioallantoic membrane model (CAMM)) and blood sampling technique in our study. 160 SPF eggs were divided into two groups with 80 eggs in each group, according to the model in which STZ was administered. When the relationship between blood glucose and blood insulin levels were examined, it was determined that there was a significantly strong negative correlation in the control group and ASM 1 group; and a significantly very strong negative correlation was found in the ASM 2 group and ASM 3 group. Our data indicate that the optimal STZ dose to create a DM model was 0.45 mg/egg and the best DM model was ASM. The second technique to be the best blood sampling technique for determining blood glucose levels. We believe that ASM can be used in DM studies and anti-DM drug studies in terms of its easebly, applicability, reproducibility and low cost.

Diameter measurements and variations of the hepatic arterial system in multidetector computed tomography images / Mediciones de diámetro y variaciones del sistema arterial hepático en imágenes de tomografía computarizada multidetector

SUMMARY: Liver plays an important role in many events such as bile production, blood filtration and metabolic functions. The liver is supplied by the hepatic arterial system. The hepatic arterial system anatomy has a variable structure and the rate of variation is high. In our study, we aimed to determine the diameters and variation of the arteries supplying the liver with multidetector computed tomography images. In this study, hepatic arterial system variations of 500 cases whose abdominal region was imaged with multi- detector computed tomography were evaluated and the diameters of the related arteries were measured. The mean diameters of classical and variational anatomy were determined in this study. According to mean measurements of classical and variational anatomy were abdominal aorta 21.95 mm, celiac artery 7.2 mm, common hepatic artery 4.3 mm, proper hepatic artery 2.93 mm, right hepatic artery 2.92 mm, left hepatic artery 2.51 mm and abdominal aorta 21.85 mm, celiac artery 6.99 mm, common hepatic artery 5.07 mm, proper hepatic artery 3.83 mm, right hepatic artery 2.87 mm ve left hepatic artery 2.09 mm respectively. When evaluated in terms of variations, 85.6 % of the cases had branching according to Type I, 14.4 % of the cases had different branching patterns. Type III (87.5 %) was the most observed variation among them. As a result of the study, it was determined that the arterial diameters vary according to the state of variation and that the arterial diameter of men are greater than that of women.

RESUMEN: El hígado juega un papel importante en diferentes eventos, tal como la producción de bilis, la filtración de sangre y las funciones metabólicas. El hígado está irrigado por el sistema arterial hepático. La anatomía del sistema arterial hepático tiene una estructura variable y la tasa de variación es alta. En nuestro estudio, nuestro objetivo fue determinar los diámetros y la variación de las arterias que irrigan el hígado con imágenes de tomografía computarizada multidetector. Se evaluaron las variaciones del sistema arterial hepático de 500 casos y se obtuvieron imágenes con tomografía computarizada de detectores múltiples abdominales y se midieron los diámetros de las arterias relacionadas. Se determinaron los diámetros medios de la anatomía clásica y variacional. Según las medidas medias de la anatomía clásica y variacional fueron aorta abdominal 21,95 mm, arteria celíaca 7,2 mm, arteria hepática común 4,3 mm, arteria hepática propia 2,93 mm, arteria hepática derecha 2,92 mm, arteria hepática izquierda arteria 2,51 mm y parte abdominal de la aorta 21,85 mm, arteria celíaca 6,99 mm, arteria hepática común 5,07 mm, arteria hepática propia 3,83 mm, arteria hepática derecha 2,87 mm y arteria hepática izquierda 2,09 respectivamente. Cuando se evaluó en términos de variaciones, el 85,6 % de los casos tenían ramificaciones según el Tipo I, el 14,4 % de los casos tenían diferentes patrones de ramificación. El tipo III (87,5 %) fue la variación más observada entre ellos. Como resultado del estudio, se determinó que los diámetros arteriales varían según el estado de variación y que el diámetro arterial de los hombres es mayor que el de las mujeres.

Effect of Pethidine Hydrochloride on the Development of Neural Tube: A Genetic Analysis Study in a Chick Embryo Model.

BACKGROUND: Neural tube defects are among the most frequent congenital abnormalities of the central nervous system. Progression of neural tube deficits is affected by hereditary predilection and environmental determinants. Pethidine (meperidine) is a fast and powerful opioid analgesic in U.S. Food and Drug Administration category C. There are reports about developmental anomalies due to this medication. The aim of this study was to investigate the effects of different doses of pethidine hydrochloride on neural tube development in a chick embryo model resembling the first month of vertebral growth in mammals. METHODS: Seventy-five specific pathogen-free eggs were incubated for 28 hours and divided into 5 groups (including the control group), each consisting of 15 eggs. Pethidine hydrochloride was administered sub-blastodermically with a Hamilton microinjector in 4 different doses. Incubation was continued until the end of the 48th hour. Subsequently, all eggs were opened, and embryos were cut from the embryonic membranes and evaluated morphologically, genetically, and histopathologically. RESULTS: Crown-rump length, somite numbers, and silver-stained nucleolar organizer region (AgNOR) number averages, and total AgNOR/nuclear area ratios decreased in a dose-dependent manner. Examination of neural tube closure revealed statistically significant differences in all experimental groups (P<0.05). Messenger RNA levels of the BRE gene were decreased in pethidine hydrochloride-exposed embryos compared with the control group. Although this downregulation was not statistically significant, this decrease was striking with a 0.422-fold change in the fifth group. CONCLUSIONS: We demonstrated that pethidine hydrochloride affects neuronal development in chicken embryos. The teratogenic mechanism of pethidine hydrochloride is unclear; therefore, further investigation is required.

Antioxidant effects of piperine in in-vivo chick embryo cataract model induced by steroids.

PURPOSE: The etiopathogenesis of steroid-induced cataracts is unknown. One hypothesis is that the higher reactive oxygen species (ROS) levels play an important role in the pathogenesis of several disorders, including the evolution of cataracts. This study investigated the antioxidant effects of piperine in our steroid-induced chick embryo lens model. METHODS: The study included 36 specific pathogen-free (SPF) fertilized eggs divided into six groups: phosphate buffer saline (PBS, pH 7.4 Saline Solution (0.9%) isotonic) group (G1), hydrocortisone succinate sodium (HC)-treated group (G2), 100 mg/kg piperine and HC treated group (G3), 50 mg/kg piperine and HC treated group (G4), 25 mg/kg piperine and HC treated group (G5), and 10 mg/kg piperine and HC treated group (G6). On the 15th day of incubation, the SPF eggs in the six groups were removed from the incubator; all were injected using insulin injectors into the chorioallantoic membrane. On day 17, all of the chick embryos were removed from the eggs and all lenses were dissected from the embryos. Cataract formation was evaluated in all lenses, and total antioxidant status (TAS), total oxidant status (TOS), reduced glutathione (GSH), and lipid peroxidation (MDA, malondialdehyde) levels were measured in all lens. RESULTS: The lenses in the G1 group had higher levels of GSH and TAS (p < 0.01), and lower levels of MDA and TOS than the G2 group (p < 0.05 and p < 0.01, respectively). Group 3 had higher levels of GSH and TAS (p < 0.001 and p < 0.001 respectively), and lower levels of MDA and TOS than the G2 group (p < 0.01 and p < 0.001, respectively). CONCLUSION: Steroid therapy causes a decrease in GSH and TAS levels and an increase in TOS and MDA levels in lens tissues, indicating increased oxidative stress. Piperine exerts its effects as an antioxidant substance, in increasing doses.