Effects of the ELF-MFs on the development of spleens of preincubated chicken embryos.

Effects of extremely low-frequency magnetic fields (ELF-MFs) on the histopathology, histomorphometry, ultrastructural changes (transmission electron microscope, TEM), apoptosis and alkaline phosphatase (ALP) activities in the spleens of preincubated white leghorn chicken embryo were investigated. Ninety fertilized eggs were divided into six groups as control (n = 15), sham (n = 15) and four experimental groups (1-4, n = 15). Eggs of experimental groups were exposed to 1.33, 2.66, 5.52 and 7.32 mTs flux intensities established in our previous published experiments and the last intensity was used for studying apoptosis and ultrastructures (TEM) of the spleens of 19-day-old chicken embryos. Eggs of control groups remained intact. Sham groups were placed inside the coil for 24 h before incubation with no exposures, then they were incubated in the same incubator (37°C, 60% humidity) for 19 days. Spleens of chicken embryos were removed, fixed in formalin (10%), and stained with H&E for histopathological and histomorphometrical surveys; TUNEL assay indicated possible change in apoptosis and TEM and biochemical studies were also carried out. Significant decreases in the sizes of the spleens of embryos of experimental groups, hyperemia, damages in spleen parenchyma, decreases in the numbers of splenic nodules, increases in the number of polymorphonuclear cells and sinusoidal spaces of spleens, significant increase in the level of ALP activities, significant increases in the numbers of apoptotic cells, deformed nuclei and swollen mitochondria were observed in experimental groups comparing with those in control and sham groups. ELF-MF created changes which could impair the immune functions.

Effects of very rapid versus vapor phase freezing on human sperm parameters.

The aim of the present study was to compare the effects of two freezing methods, vapor phase and very rapid freezing, with and without cryoprotectant on semen parameters in men with normal semen criteria. Cryopreservation was done on semen samples from 31 men by two methods of vapor phase freezing and very rapid freezing, with and without Test Yolk buffered glycerol (TYBG) as cryoprotectant. The motility, viability, acrosome and DNA integrity were evaluated on fresh and post-thaw samples. Post-thaw sperm progressive motility was significantly higher in the presence of TYBG in the vapor phase cryopreservation (%6.30 ± 3.74) compared with the very rapid freezing method (%2.2 ± 1.97 and %4.00 ± 2.42 in the presence and absence of TYBG, respectively). There was no significant difference in viability, acrosome status and DNA integrity between two methods in presence or absence of TYBG. The very rapid freezing method in the absence of TYBG showed better sperm motility but viability, acrosome and DNA integrity were similar to the presence of TYBG. The results show that cryopreservation of human spermatozoa together with seminal plasma by using vapor phase method is better than very rapid freezing method to preserve sperm progressive motility; however very rapid freezing method is quick and simple and do not require special cryoprotectant. It can be used for cryopreservation of small number of spermatozoa in IVF centers.

Quinazolinone exposure induces apoptosis and changes expressions of Fas/FasL and C-Flip in embryonic mice testicles.

Quinazolinones represent a class of sedative and anticancer drugs. Quinazolinones-based compounds have ability to suppress prostate tumor growth via apoptosis. Apoptosis is very common in embryos and adults of normal and injured mammalian testes. Effects a new derivative of quinazolinone (4(3H) quinazolinone-2-ethyl-2-phenyl ethyl (QEPE)), on the testis of Balb/C mice embryos were investigated. QEPE was able to reduce number of germ cells and diameter of seminiferous tubules. TUNEL assay analysis indicated that reduction correlated with an increase in the number of apoptotic cell. Furthermore, electron microscope observations confirmed typical apoptotic morphologies characterized by chromatin fragmentation. Finally, RT-PCR analysis showed QEPE increases the levels of Fas/Fasl and decreases C-Flip mRNAs in the testis of exposed embryos.

Effects of sinusoidal electromagnetic fields on histopathology and structures of brains of preincubated white Leghorn chicken embryos.

There are several reports indicating linkages between exposures to 50-60 Hz electromagnetic fields and abnormalities in the early stages of chicken embryonic development. Based on our previous published research carried out at the Department of Animal Sciences, Faculty of Biological Sciences, Shahid Beheshti University, effects of sinusoidal electromagnetic fields on histopathology and structures of brains of preincubated white leghorn hen eggs were investigated. Three hundred healthy fresh fertilized eggs (55-65 gr) were divided into three groups of experimental (n = 50), control (n = 75), and sham (n = 75). Experimental eggs (inside the coil) were exposed to 3 different intensities of 1.33, 2.66, and 7.32 mT and sham groups were located inside the same coil with no exposure, for 24 h before incubation. Control, sham, and experimental groups were all incubated in an incubator (38 ± 0.5(°)C, 60% humidity) for 14 days. 14-day old chicken embryos were removed by C-sections, and the brains of all embryos of all groups were fixed in formalin(10%), stained with H&E and TUNEL assay, for studying the histopatholog and process of apoptosis. The brains of other embryos were prepared for Scanning Electeron Microscope. Results showed electromagnetic fields have toxic effects on brain cells by increasing the number of apoptotic cells and degeneration of brains' tissues of exposed chicken embryos. These findings suggest that the electromagnetic fields induce brain damages at different levels.

Effects of 50 Hz electromagnetic fields on the histology, apoptosis, and expression of c-Fos and ß-catenin on the livers of preincubated white Leghorn chicken embryos.

Reports have demonstrated occurrences of abnormalities in the early stages of chicken embryonic development due to the exposure to electromagnetic fields (EMFs). This article was designed to investigate the effects of sinusoidal EMF on the histopathology, apoptosis, and expressions of c-Fos and ß-Catenin genes of the livers of preincubated White Leghorn chicken embryos, based on our published experiments. 300 healthy, fresh fertilized eggs were divided into control (n = 70), sham (n = 70), and four experimental (1-4,days 13, 14, 5, and 19, n = 40) groups. Experimental eggs were exposed to the most effective intensity in a coil with 7.32 mT density, and sham groups were also located in the same coil with no exposure, both for 24 h before incubation. Control, sham, and experimental groups were then incubated in an incubator (37°C, humidity 60%) for 13,14,15, and 19 days. Livers of 13-15 and 19 day-old chicken embryos were removed by C-section and fixed in formalin (10%), stained with Hematoxylin-Eosin and TUNEL for histopathological and apoptosis studies. Others were used for investigating c-Fos and ß-Catenin expressions, using RT-PCR. Results showed extensive hemorrhages all over the chicken embryos' bodies and livers, more lymphoid tissues, disturbed parenchymal tissues, sinusoid denaturation, vesiculizad cytoplasm, an increase in the number of apoptotic cells, and a decrease on the levels of expressions of c-Fos and ß-Catenin genes in experimental groups of 1-4, comparing control and sham groups.

Histopathological and ultrastructural studies on the effects of electromagnetic fields on the liver of preincubated white Leghorn chicken embryo.

There are several reports indicating a linkage between exposures to 50-60 Hz electromagnetic fields and abnormalities in the early stages of embryonic development of chicken embryos. The present study was designed to demonstrate whether electromagnetic fields could be an environmental factor invoking histopathological and ultra-structural changes in livers of preincubated chicken embryos exposed to EMFs. Following other researchers and our previous results from different groups of Developmental Biology at the Animal Sciences, Faculty of Biological Sciences, Shahid-Beheshti University, effects of most effective intensities (1.33, 2.66, 5.52, and 7.32 mT) of electromagnetic fields (EMFs, 50 Hz ) on livers of pre-incubated white leghorn chicken embryos were investigated . 150 healthy, fresh, and fertilized eggs (55-65 gr) were divided into 6 groups of experimental(1-4, n = 30), control (n = 60), and sham (n = 50). Experimental eggs (inside coil) were exposed to 4 different intensities (1.33, 2.66, 5.52, and 7.32 mT). Sham groups were located inside same coil, with no exposure, for 24 h before incubation. Control, sham, and experimental groups (1-4) were then incubated in an incubator (38 +/- 0.5 degrees C, 60% humidity) for 17 days. At the end of this period, livers of experimental, sham, and control groups were processed for light and transmission electrom microscopes (TEM and SEM) studies. So, livers of 17-day old chicken embryos were removed by C-sections, fixed in formalin 10%, stained with H&E and reticulin, and studied under light microscope. Others were prepared for electron microscopes (TEM and SEM) investigations. Morphological observations indicated exencephalic embryos, embryos with asymmetrical faces, crossed beak, shorter upper beak, deformed hind limbs, gastroschesis, anophthalmia, and microphthalmia. H&E and reticulin stainings, TEMS, and SEMs studies indicated EMFs would create hepato-cytes with fibrotic bands, severe steatohepatitis, vacuolizations, swollen and extremely electron-dense mitochondria, reduced invisible cristae, crystalized mitochondria with degenerated cristae, myelin-like figures, macrophages engulfing adjacent cells, dentated nuclei, nuclei with irregular envelopes, degenerated hepatocytes, abnormal lipid accumulations, lipid droplets pushing hepatocytes' nuclei to the corner of the cells, abundant cellular infiltrations cellular infiltrations inside sinusoid and around central veins, disrupted reticulin plexus, and release of chromatin into cytosol,, with partially regular water layers. An elevated oxyradical generation and, subsequently, cell membrane disruptions were the reasons for electromagnetic fields inducing cell damages.

Development of preincubated chicken eggs following exposure to 50 Hz electromagnetic fields with 1.33-7.32 mT flux densities.

The effects of applying extremely low-frequency (50 Hz) electromagnetic fields (ELF-EMF) for 24 hr and different densities (1.33-7.32 mT) were examined on healthy, freshly fertilized white leghorn chicken eggs (55-65 g). Results showed no increase in the rate of abnormalities in exposed groups, but were only significant in 4.19, 5.32 and 5.86, 6.65 mT densities. Alizarin red S and alcian blue 8GX staining showed some embryos with extra ribs, defects in ribs and vertebrae, anuria and abnormal beaks. Study of egg weight, after 9 days of incubation, showed no significant differences between control, sham-exposed and experimental groups. Analysis of crown-rump, beak-occipital length and weight of embryo, showed significant decrease in weight at 4.39 and 5.52 mT intensities, comparing with control and sham-exposed groups. These results revealed that 50 Hz electromagnetic fields can even induce developmental alterations in preincubated chick embryos and confirm that its strength could be a determinant factor for the embryonic response to extremely low frequency EMFs (window effects) prior to incubation.