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1.  NEUROBEHAVIORAL TERATOGENICITY OF SARIN IN AN AVIAN MODEL 
Neurotoxicology and teratology  2009;31(6):406-412.
Nerve gas organophosphates like sarin are likely to be used in urban terrorism, leading to widespread exposures of pregnant women and young children. Here, we established a model for sarin neurobehavioral teratogenicity in the developing chick so as to explore the consequences of apparently subtoxic sarin exposure and the mechanisms underlying synaptic and behavioral deficits. Chicken eggs were injected with sarin (2, 6 and 12 μg/kg) on incubation days 2 and 6, treatments that did not decrease hatching and did not evoke dysmorphology. After hatching the chicks were tested for filial imprinting and neurochemical markers known to be critical for imprinting. Imprinting was reduced at 2 and 6 μg/kg but not at the highest dose. Acetylcholinesterase and choline acetyltransferase were unaffected but sarin reduced the concentration of the high-affinity choline transporter, the rate-limiting factor in acetylcholine utilization. The concentration of PKC isoforms was assessed in the imprinting-related intermediate part of the medial hyperstriatum ventrale, the region most closely associated with cholinergic function in imprinting behavior. Sarin reduced the concentration of all isoforms (α, β, γ) with a similar, biphasic dose-response curve to that seen for behavioral performance, a relationship noted in previous work with organophosphate pesticides. Our results indicate that otherwise subtoxic exposures to sarin produce neurodevelopmental deficits; since we utilized a chick model, which is devoid of maternal confounds that are present in mammalian development, the adverse effects of sarin are mediated directly in the developing organism.
doi:10.1016/j.ntt.2009.07.007
PMCID: PMC2761973  PMID: 19660543
Acetylcholine systems; Brain development; Chlorpyrifos; Cholinesterase; Developing chick; Organophosphates; Serotonin systems
2.  Nano-Nutrition of Chicken Embryos. The Effect of in Ovo Administration of Diamond Nanoparticles and l-Glutamine on Molecular Responses in Chicken Embryo Pectoral Muscles 
It has been demonstrated that the content of certain amino acids in eggs is not sufficient to fully support embryonic development. One possibility to supply the embryo with extra nutrients and energy is in ovo administration of nutrients. Nanoparticles of diamond are highly biocompatible non-toxic carbonic structures, and we hypothesized that bio-complexes of diamond nanoparticles with l-glutamine may affect molecular responses in breast muscle. The objective of the investigation was to evaluate the effect of diamond nanoparticle (ND) and l-glutamine (Gln) on expression of growth and differentiation factors of chicken embryo pectoral muscles. ND, Gln, and Gln/ND solutions (50 mg/L) were injected into fertilized broiler chicken eggs at the beginning of embryogenesis. Muscle tissue was dissected at day 20 of incubation and analysed for gene expression of FGF2, VEGF-A, and MyoD1. ND and especially Gln/ND up-regulated expression of genes related to muscle cell proliferation (FGF2) and differentiation (MyoD1). Furthermore, the ratio between FGF2 and MyoD1 was highest in the Gln/ND group. At the end of embryogenesis, Gln/ND enhanced both proliferation and differentiation of pectoral muscle cells and differentiation dominated over proliferation. These preliminary results suggest that the bio-complex of glutamine and diamond nanoparticles may accelerate growth and maturation of muscle cells.
doi:10.3390/ijms141123033
PMCID: PMC3856104  PMID: 24264045
chicken embryo; myogenesis; diamond nanoparticles; l-glutamine; gene expression
3.  Expression of mRNA for 3HADH in manipulated embryos to produce germline chimeric chickens 
Cytotechnology  2009;60(1-3):165-168.
Germline chimeric chickens were produced by the transfer of primordial germ cells (PGCs) or blastoderm cells. The hatchability of eggs produced by transfer of exogenous PGCs is usually low. The purpose of the present study was investigated to express (3-hydroxyacyl CoA dehydrogenase) 3HADH which is a limiting enzyme in the beta-oxidation of fatty acids for hatching energy. Manipulations of both donor and recipient eggshells were as follows. A window approximately 10 mm in diameter was opened at the pointed end of the eggs at stage 12–15 days incubation. Donor PGCs, taken from the blood vessels of donor embryos from fertilized eggs at the same stage of development, were injected into the blood vessels of recipient embryos. The muscles of chicks in the eggs with transferred PGCs were removed after 20 days of incubation. A cDNA was prepared from the total RNA. The expression of 3HADH in the manipulated embryos was investigated using real-time PCR analysis. Real-time PCR analysis showed that expression of 3HADH was reduced in the muscles of manipulated embryos.
doi:10.1007/s10616-009-9232-x
PMCID: PMC2780556  PMID: 19856125
Manipulated embryo; PGCs; 3HADH; Hatch
4.  Ethanol- and/or Taurine-Induced Oxidative Stress in Chick Embryos 
Journal of Amino Acids  2013;2013:240537.
Because taurine alleviates ethanol- (EtOH-) induced lipid peroxidation and liver damage in rats, we asked whether exogenous taurine could alleviate EtOH-induced oxidative stress in chick embryos. Exogenous EtOH (1.5 mmol/Kg egg or 3 mmol/Kg egg), taurine (4 μmol/Kg egg), or EtOH and taurine (1.5 mmol EtOH and 4 μmol taurine/Kg egg or 3 mmol EtOH and 4 μmol taurine/Kg egg) were injected into fertile chicken eggs during the first three days of embryonic development (E0–2). At 11 days of development (midembryogenesis), serum taurine levels and brain caspase-3 activities, homocysteine (HoCys) levels, reduced glutathione (GSH) levels, membrane fatty acid composition, and lipid hydroperoxide (LPO) levels were measured. Early embryonic EtOH exposure caused increased brain apoptosis rates (caspase-3 activities); increased brain HoCys levels; increased oxidative-stress, as measured by decreased brain GSH levels; decreased brain long-chain polyunsaturated levels; and increased brain LPO levels. Although taurine is reported to be an antioxidant, exogenous taurine was embryopathic and caused increased apoptosis rates (caspase-3 activities); increased brain HoCys levels; increased oxidative-stress (decreased brain GSH levels); decreased brain long-chain polyunsaturated levels; and increased brain LPO levels. Combined EtOH and taurine treatments also caused increased apoptosis rates and oxidative stress.
doi:10.1155/2013/240537
PMCID: PMC3628655  PMID: 23606945
5.  Transfer of blood containing primordial germ cells between chicken eggs development of embryonic reproductive tract 
Cytotechnology  2007;56(1):27-32.
The present study was carried out to investigate development of recipient chicken embryonic reproductive tracts which are transferred chicken primordial germ cells (PGCs). It is thought that differentiation of PGCs is affected by the gonadal somatic cells. When female PGCs are transferred to male embryos, it is possible that they differentiate to W-spermatogonia. However, the relationship development between PGCs and gonads has not been investigated. At stage 12–15 of incubation of fertilized eggs, donor PGCs, which were taken from the blood vessels of donor embryos, were injected into the blood vessels of recipient embryos. The gonads were removed from embryos that died after 16 days of incubation and from newly hatched chickens and organs were examined for morphological and histological features. The survival rate of the treated embryos was 13.6% for homo-sexual transfer of PGCs (male PGCs to male embryo or female PGCs to female embryo) and 28.9% for hetero-sexual transfer PGCs (male PGCs to female embryo or female PGCs to male embryo) when determined at 15 days of incubation. The gonads of embryos arising from homo-sexual transfer appeared to develop normally. In contrast, embryos derived from hetero-sexual transfer of PGCs had abnormal gonads as assessed by histological observation. These results suggest that hetero-sexual transfer of PGCs may influence gonadal development early-stage embryos.
doi:10.1007/s10616-007-9096-x
PMCID: PMC2151963  PMID: 19002838
Chicken; Embryo; Gonad; PGCs; Transfer
6.  Effects of power frequency electromagnetic fields on melatonin and sleep in the rat 
Emerging Health Threats Journal  2012;5:10.3402/ehtj.v5i0.10904.
Background
Studies investigating the effect of power frequency (50–60 Hz) electromagnetic fields (EMF) on melatonin synthesis in rats have been inconsistent with several showing suppression of melatonin synthesis, others showing no effect and a few actually demonstrating small increases. Scant research has focused on the ensuing sleep patterns of EMF exposed rats. The present study was designed to examine the effects of extremely low power frequency electromagnetic fields (EMF) on the production of melatonin and the subsequent sleep structure in rats.
Methods
Eighteen male Sprague-Dawley rats were exposed to a 1000 milligauss (mG) magnetic field for 1 month. Urine was collected for the final 3 days of the exposure period for analysis of 6-sulphatoxymelatonin, the major catabolic product of melatonin found in urine. Subsequent sleep was analyzed over a 24-hour period.
Results
Melatonin production was mildly increased in exposed animals. Although there were no statistically significant changes in sleep structure, exposed animals showed slight decreases in REM (rapid eye movement) sleep as compared to sham (non-exposed) animals.
Conclusions
Power frequency magnetic fields induced a marginally statistically significant increase in melatonin levels in exposed rats compared to control. Subsequent sleep analysis indicated little effect on the sleep architecture of rats, at least not within the first day after 1 month's continuous exposure. Varying results in the literature are discussed and future research suggested.
doi:10.3402/ehtj.v5i0.10904
PMCID: PMC3334267  PMID: 22529876
electromagnetic fields; 6-sulphatoxymelatonin; Sprague-Dawley; sleep
7.  Transmission OF Campylobacter coli in chicken embryos 
Brazilian Journal of Microbiology  2012;43(2):535-543.
Campylobacter coli is an important species involved in human cases of enteritis, and chickens are carriers of the pathogen mainly in developing country. The current study aimed to evaluate the transmission of C. coli and its pathogenic effects in chicken embryos. Breeder hens were inoculated intra-esophageally with C. coli isolated from chickens, and their eggs and embryos were analyzed for the presence of bacteria using real-time PCR and plate culture. The viability of embryos was verified. In parallel, SPF eggs were inoculated with C. coli in the air sac; after incubation, the embryos were submitted to the same analysis as the embryos from breeder hens. In embryos and fertile eggs from breeder hens, the bacterium was only identified by molecular methods; in the SPF eggs, however, the bacterium was detected by both techniques. The results showed no relationship between embryo mortality and positivity for C. coli in the embryos from breeder hens. However, the presence of bacteria is a cause of precocious mortality for SPF embryos. This study revealed that although the vertical transmission is a possible event, the bacteria can not grow in embryonic field samples.
doi:10.1590/S1517-83822012000200014
PMCID: PMC3768811  PMID: 24031861
Campylobacter coli; Viability; Transmission; Breeder hens
8.  Altered neurochemical profile after traumatic brain injury: 1H-MRS biomarkers of pathological mechanisms 
Specific neurochemicals measured with proton magnetic resonance spectroscopy (1H-MRS) may serve as biomarkers of pathological mechanism in the brain. We used high field in vivo 1H-MRS to measure a detailed neurochemical profile after experimental traumatic brain injury (TBI) in rats. We characterized neurochemical changes in the contused cortex and the normal-appearing perilesional hippocampus over a time course from 1 hour to 2 weeks after injury. We found significant changes in 19 out of 20 neurochemicals in the cortex, and 9 out of 20 neurochemicals in the hippocampus. These changes provide evidence of altered cellular metabolic status after TBI, with specific compounds proposed to reflect edema, excitotoxicity, neuronal and glial integrity, mitochondrial status and bioenergetics, oxidative stress, inflammation, and cell membrane disruption. Our results support the utility of 1H-MRS for monitoring cellular mechanisms of TBI pathology in animal models, and the potential of this approach for preclinical evaluation of novel therapies.
doi:10.1038/jcbfm.2012.114
PMCID: PMC3519407  PMID: 22892723
animal models; brain trauma; cell death mechanisms; MR spectroscopy; neurochemistry
9.  Micro-magnetic resonance imaging of avian embryos 
Journal of Anatomy  2007;211(6):798-809.
Chick embryos are useful models for probing developmental mechanisms including those involved in organogenesis. In addition to classic embryological manipulations, it is possible to test the function of molecules and genes while the embryo remains within the egg. Here we define conditions for imaging chick embryo anatomy and for visualising living quail embryos. We focus on the developing limb and describe how different tissues can be imaged using micro-magnetic resonance imaging and this information then synthesised, using a three-dimensional visualisation package, into detailed anatomy. We illustrate the potential for micro-magnetic resonance imaging to analyse phenotypic changes following chick limb manipulation. The work with the living quail embryos lays the foundations for using micro-magnetic resonance imaging as an experimental tool to follow the consequences of such manipulations over time.
doi:10.1111/j.1469-7580.2007.00825.x
PMCID: PMC2375841  PMID: 18045352
anatomy; avian; chick; embryo; limb development; using micro-magnetic resonance imaging; magnetic resonance imaging; quail
10.  Micro-magnetic resonance imaging of avian embryos 
Journal of Anatomy  2007;211(6):798-809.
Chick embryos are useful models for probing developmental mechanisms including those involved in organogenesis. In addition to classic embryological manipulations, it is possible to test the function of molecules and genes while the embryo remains within the egg. Here we define conditions for imaging chick embryo anatomy and for visualising living quail embryos. We focus on the developing limb and describe how different tissues can be imaged using micro-magnetic resonance imaging and this information then synthesised, using a three-dimensional visualisation package, into detailed anatomy. We illustrate the potential for micro-magnetic resonance imaging to analyse phenotypic changes following chick limb manipulation. The work with the living quail embryos lays the foundations for using micro-magnetic resonance imaging as an experimental tool to follow the consequences of such manipulations over time.
doi:10.1111/j.1469-7580.2007.00825.x
PMCID: PMC2375841  PMID: 18045352
anatomy; avian; chick; embryo; limb development; using micro-magnetic resonance imaging; magnetic resonance imaging; quail
11.  Otic Lesions and Congenital Hypothyroidism in the Developing Chick* 
Journal of Clinical Investigation  1967;46(11):1828-1839.
In an effort to elucidate the relation, if any, between thyroid abnormality and congenital deafness in Pendred's syndrome, an experiment was designed to study the effects of hypothyroidism on middle and inner ear hearing structures, including the auditory nerve and its central projection, in developing chick embryos. Propylthiouracil (PTU), 2 mg, was injected into the albumin of fertile chick eggs on the 10th incubation day. Single doses of L-thyroxine (range 1-100μg) were inoculated in a similar manner, either alone or with PTU. Control inocula included sterile saline or water. After hatching, each chick was examined for obvious malformations. The thyroid glands, middle and inner ear mechanisms, auditory nerve, and brainstem were studied grossly and with different histologic staining techniques. When compared to controls, chicks exposed to PTU on their 10th incubation day exhibited: increased mortality, delayed hatching, reduced size, incomplete yolk sac absorption, and death within 5 days unless exogenous thyroid hormone was provided in the first 24-48 hr after hatching. Specific, consistent, morphologic alterations were observed in their thyroid glands as well as in the sensory hair cells of the acoustic papilla and cells of the spiral ganglion of the cochlea. Our data also indicate that if 50-75 μg of L-thyroxine is given simultaneously with (or as long as 120 hr after) the PTU injection on the 10th incubation day, one cannot detect the gross defects, marked thyroid lesions, or abnormal histology in cells of the cochlea and its ganglion. A relationship between embryonic thyroid gland function and the hearing mechanism of the chick embryo is suggested.
Images
PMCID: PMC292933  PMID: 6070327
12.  Power-line frequency electromagnetic fields do not induce changes in phosphorylation, localization, or expression of the 27-kilodalton heat shock protein in human keratinocytes. 
Environmental Health Perspectives  2003;111(3):281-288.
The linkage of the exposure to the power-line frequency (50-60 Hz) electromagnetic fields (EMF) with human cancers remains controversial after more than 10 years of study. The in vitro studies on the adverse effects of EMF on human cells have not yielded a clear conclusion. In this study, we investigated whether power-line frequency EMF could act as an environmental insult to invoke stress responses in human keratinocytes using the 27-kDa heat shock protein (HSP27) as a stress marker. After exposure to 1 gauss (100 micro T) EMF from 20 min to 24 hr, the isoform pattern of HSP27 in keratinocytes remained unchanged, suggesting that EMF did not induce the phosphorylation of this stress protein. EMF exposure also failed to induce the translocation of HSP27 from the cytoplasm to the nucleus. Moreover, EMF exposure did not increase the abundance of HSP27 in keratinocytes. In addition, we found no evidence that EMF exposure enhanced the level of the 70-kDa heat shock protein (HSP70) in breast or leukemia cells as reported previously. Therefore, in this study we did not detect any of a number of stress responses in human keratinocytes exposed to power-line frequency EMF.
PMCID: PMC1241383  PMID: 12611655
13.  Early embryogenesis in zebrafish is affected by bisphenol A exposure 
Biology Open  2013;2(5):466-471.
Summary
Exposure of a developing embryo or fetus to endocrine disrupting chemicals (EDCs) has been hypothesized to increase the propensity of an individual to develop a disease or dysfunction in his/her later life. Although it is important to understand the effects of EDCs on early development in animals, sufficient information about these effects is not available thus far. This is probably because of the technical difficulties in tracing the continuous developmental changes at different stages of mammalian embryos. The zebrafish, an excellent model currently used in developmental biology, provides new insights to the field of toxicological studies. We used the standard whole-mount in situ hybridization screening protocol to determine the early developmental defects in zebrafish embryos exposed to the ubiquitous pollutant, bisphenol A (BPA). Three stages (60–75% epiboly, 8–10 somite, and prim-5) were selected for in situ screening of different molecular markers, whereas BPA exposure altered early dorsoventral (DV) patterning, segmentation, and brain development in zebrafish embryos within 24 hours of exposure.
doi:10.1242/bio.20134283
PMCID: PMC3654264  PMID: 23789094
Bisphenol A; Embryogenesis; Zebrafish
14.  Isolation and cultivation of bovine ephemeral fever virus in chickens and chicken embryos. 
The Journal of Hygiene  1978;81(1):1-7.
Unadapted bovine ephemeral fever (BEF) virus was isolated from cattle blood after intravenous inoculation into chicken embryos. Infected embryos died or hatched as abnormal chickens. The chick embryo was slightly less sensitive to unadapted BEF virus than were Vero cell cultures, but the use of embryos avoids the several blind passages that are required to isolate BEF virus in unweaned mice. Chick embryos were considerably less efficient than Vero cell culture or unweaned mice in detecting Vero cell-adapted and mouse-adapted BEF virus respectively. Viraemia was demonstrated in chicken embryos at 1-4 days and in one-day-old chickens at 1-3 days after intravenous inoculation of BEF virus. BEF virus was demonstrated by isolation and by immunofluoresence in heart, brain, lung and liver of chicken embryos at 1-5 days and in lung and liver of one-day-old chickens at 1-2 days, after intravenous inoculation. The isolated viruses were confirmed as BEF virus by neutralization with immune mouse ascitic fluid. BEF neutralizing antibodies were produced in 4-week-old and adult chickens after intravenous inoculation with BEF virus.
PMCID: PMC2129751  PMID: 357649
15.  An In vitro Study on Chick Somite Ability to Express Cerberus, Chordin, FGF8, Follistatin, and Noggin Transcripts 
Background
In vitro simulation of developmental processes is an invaluable tool to shed light on the intrinsic mechanism of developmental biosystems such as central nervous system in mammals. Chick somites have been used to simulate the neural differentiation of human neural progenitor cells. In the present study, we aimed to indicate whether somites have the ability to express required neural differentiation factors at mRNA level.
Methods
Chick embryos were isolated from the yolk surface of the fertilized eggs and somites were subsequently isolated from embryos under a dissecting microscope. Total RNA of the somites was extracted and RT-PCR carried out with specific primers of cerberus, chordin, FGF8, follistatin and noggin.
Results
Data showed that five aforementioned factors were co-expressed after 7 days in vitro by somites.
Conclusion
We concluded that neural induction property of somites appeared by production of required neural differentiation factors including cerberus, chordin, FGF8, follistatin and noggin.
PMCID: PMC4009094  PMID: 24834314
Cerberus protein; Chordin; FGF8 protein; Follistatin; Noggin protein; Somites
16.  Effect of prenatal chronic noise exposure on the growth and development of body and brain of chick embryo 
Context:
Noise acts as an environmental stressor as has been demonstrated by an increased brain acetyl cholinesterase activity as well as elevated plasma corticosterone and adrenocorticotropic hormone levels. Noise can lead to neurodegenerative changes in the brain and in the ear.
Aim:
This study was undertaken to investigate the effect of chronic noise on growth and development during the sensitive period of embryonic life.
Materials and Methods:
In this study, we analyzed the body weight, brain weight and brain size following prenatal chronic noise exposure. Fertilized eggs of domestic chicks were exposed to chronic excessive acoustic stimulation with frequency of the sound ranging from 30 to 3000 Hz with a peak at 2700 Hz was given at 110 dB sound pressure level from embryonic day (E) 10 until hatching.
Results:
An appreciable decrease in body weight, brain weight and brain size was evident in the experimental group exposed to noise. A generalized decrease in the neuronal nuclear size and increase in the density of neurons was also observed.
Conclusion:
These observations could be an indicator of growth and developmental retardation following exposure to noise.
doi:10.4103/2229-516X.125666
PMCID: PMC3931211  PMID: 24600569
Development; environmental stressor; growth; noise
17.  Effects of short term exposure of eggs to magnetic field before incubation on hatchability and post-hatch performance of meat chickens 
The effects of exposing meat-type breeder eggs to magnetic field (MF) before incubation on hatchability traits (percents of hatchability and hatchability failures of eggs), chick weight at hatch, and post-hatch performance (weight gain, feed intake, and feed conversion ratio (FCR)) from 1 to 39 d of age were investigated. Eggs from a Ross flock at 38 weeks of age were exposed to MF of 18 Gauss (1.8 mT) at 50 Hz for 0, 15, 30, 45, 60, and 75 min (MF0, MF15, MF30, MF45, MF60, and MF75) before incubation.
Exposing eggs to MF did not influence hatchability of eggs and chick weight at hatch. However, chickens of MF60 and MF75 treatments had lower weight gain and feed intake than those of the non-exposed treatment at 39 d of age. MF exposure of eggs did not influence FCR of chickens between 1 and 21 d of age, but tended to increase FCR, albeit non-significantly, between 22 and 39 d of age.
It is concluded that exposing meat-type breeder eggs to MF of 18 Gauss (1.8 mT) at 50 Hz for up to 75 min did not influence hatchability traits and chick weight at hatch. However, MF exposure of eggs for 60 and 75 min reduced body weight gain and feed intake of chickens over the 39-d experimental period.
doi:10.1016/j.sjbs.2011.06.004
PMCID: PMC3730795  PMID: 23961150
Magnetic field; Chickens; Hatchability; Post-hatch performance
18.  Laboratory infection of chicken eggs with Campylobacter jejuni by using temperature or pressure differentials. 
Fertile chicken eggs were infected in our laboratory with Campylobacter jejuni suspensions by using temperature or pressure differential methods of inoculation. After 2 days of incubation, over 90% of the eggs carried C. jejuni when iron was present in the inoculum. This percentage declined rapidly until by day 8, less than 10% of the eggs were detectably infected. However, up to 11% of hatched, healthy chicks carried C. jejuni in their intestinal tracts. The isolated organisms were of the same serotype as the initial inoculum. C. jejuni was recovered without difficulty when the intestinal tracts of chicks were enriched, but recovery from early dead-in-shell or infertile eggs was poor. This poor recovery and the rapid decline of C. jejuni after 2 days of egg incubation suggest that the vibrio is sensitive to some part of the incubating egg or to the temperature of prolonged incubation. It was impossible to predict which eggs would yield infected chicks on the basis of the number of organisms taken up by each egg, and no correlation existed between the number of organisms taken up and the efficiency of the hatch, i.e., the hatch ratio. If iron was omitted from the inoculum broth, the egg infection rate at day 2 was lower.
PMCID: PMC241748  PMID: 4015086
19.  NEUROCHEMICAL ALTERATIONS IN SPINOCEREBELLAR ATAXIA TYPE 1 AND THEIR CORRELATIONS WITH CLINICAL STATUS 
Robust biomarkers of neurodegeneration are critical for testing of neuroprotective therapies. The clinical applicability of such biomarkers requires sufficient sensitivity to detect disease in individuals. Here we tested the sensitivity of high field (4 tesla) proton magnetic resonance spectroscopy (1H MRS) to neurochemical alterations in the cerebellum and brainstem in spinocerebellar ataxia type 1 (SCA1). We measured neurochemical profiles that consisted of 10–15 metabolite concentrations in the vermis, cerebellar hemispheres and pons of patients with SCA1 (N=9) and healthy controls (N=15). Total NAA (N-acetylaspartate + N-acetylaspartylglutamate, tNAA) and glutamate were lower and glutamine, myo-inositol and total creatine (creatine + phosphocreatine, tCr) were higher in patients relative to controls, consistent with neuronal dysfunction/loss, gliotic activity and alterations in glutamate-glutamine cycling and energy metabolism. Changes in tNAA, tCr, myo-inositol and glutamate levels were discernible in individual spectra and the tNAA/myo-inositol ratio in the cerebellar hemipheres and pons differentiated the patients from controls with 100% specificity and sensitivity. In addition, tNAA, myo-inositol and glutamate levels in the cerebellar hemispheres and the tNAA and myo-inositol levels in the pons correlated with ataxia scores (Scale for the Assessment and Rating of Ataxia, SARA). Two other biomarkers measured in the cerebrospinal fluid (CSF) of a subset of the volunteers (F2-isoprostanes as a marker of oxidative stress and glial fibrillary acidic protein (GFAP) as a marker of gliosis) were not different between patients and controls. These data demonstrate that 1H MRS biomarkers can be utilized to non-invasively assess neuronal and glial status in individual ataxia patients.
doi:10.1002/mds.23067
PMCID: PMC2916651  PMID: 20310029
SCA1; MRS; ataxia; cerebellum; neurochemical profile
20.  Histopathology and cholinergic assessment of Pterocarya fraxinifolia on chicken embryo 
Interdisciplinary Toxicology  2009;2(4):254-256.
There are no reports of toxicological studies of Pterocarya fraxinifolia. The leaves are used for fishing, which also an anesthetic agent. Currently, many drugs utilized in anesthesia practice are modified cholinergic transmission and acetylcholine esterase inhibitors; these are parts of anaesthetic pharmacy. Therefore, cholinergic assessment was surveyed in chicken embryo, which Pterocarya fraxinifolia extractes were injected in 0.1, 1 and 10 mg concentration at day 4 of incubation. Serum and brain cholinesterase were analyzed on day 20 of incubation. The signs were not due to the changes of cholinesterase activity. In histopathology examination, massive necrosis was observed in the spinal cord. Other tissues such as heart, kidneys, skeletal bones and muscles, trachea and lungs, digestive system and endocrine glands were completely developed. This data suggests that the spinal cord is a target organ of the bioactive component of this plant.
doi:10.2478/v10102-009-0023-1
PMCID: PMC2984111  PMID: 21217863
Pterocarya fraxinifolia; toxicity tests; cholinergic assessment; chicken embryotoxicity
21.  TRANSPLANTABILITY OF TISSUES TO THE EMBRYO OF FOREIGN SPECIES  
Inoculation of the Jensen rat sarcoma into the developing chick embryo gives a rapidly growing tumor at the site of inoculation, whether in the membranes or in the body of the chick itself. These tumors by transfer from embryo to embryo can be kept going for as long as forty-six days, and perhaps indefinitely in the foreign species. The rat cells show no morphological change even after a very long dependence. Their biological characters are also retained, as is shown by the fact that the cells when replanted in the rat, after a prolonged sojourn in the chick, will produce a rapidly growing sarcoma of the Jensen type. These rat tissues grown for long periods in the chick show no adaptation to the new species, being destroyed even more rapidly when placed in the adult chicken than cells taken directly from the rat. Morphologically the cells retain a close resemblance to those in the original tumor. Other tissues grown in chick embryo are various embryonic cells from the chicken, mouse, and rat, the Ehrlich sarcoma and chondroma of the mouse, a mammary carcinoma of the mouse, the Flexner-Jobling adenocarcinoma of the rat, and a human sarcoma.
PMCID: PMC2125042  PMID: 19867659
22.  The therapeutic effect of a pulsed electromagnetic field on the reproductive patterns of male Wistar rats exposed to a 2.45-GHz microwave field 
Clinics  2011;66(7):1237-1245.
INTRODUCTION:
Environmental exposure to man-made electromagnetic fields has been steadily increasing with the growing demand for electronic items that are operational at various frequencies. Testicular function is particularly susceptible to radiation emitted by electromagnetic fields.
OBJECTIVES:
This study aimed to examine the therapeutic effects of a pulsed electromagnetic field (100 Hz) on the reproductive systems of male Wistar rats (70 days old).
METHODS:
The experiments were divided into five groups: microwave sham, microwave exposure (2.45 GHz), pulsed electromagnetic field sham, pulsed electromagnetic field (100 Hz) exposure, and microwave/pulsed electromagnetic field exposure. The animals were exposed for 2 hours/day for 60 days. After exposure, the animals were sacrificed, their sperm was used for creatine and caspase assays, and their serum was used for melatonin and testosterone assays.
RESULTS:
The results showed significant increases in caspase and creatine kinase and significant decreases in testosterone and melatonin in the exposed groups. This finding emphasizes that reactive oxygen species (a potential inducer of cancer) are the primary cause of DNA damage. However, pulsed electromagnetic field exposure relieves the effect of microwave exposure by inducing Faraday currents.
CONCLUSIONS:
Electromagnetic fields are recognized as hazards that affect testicular function by generating reactive oxygen species and reduce the bioavailability of androgen to maturing spermatozoa. Thus, microwave exposure adversely affects male fertility, whereas pulsed electromagnetic field therapy is a non-invasive, simple technique that can be used as a scavenger agent to combat oxidative stress.
doi:10.1590/S1807-59322011000700020
PMCID: PMC3148471  PMID: 21876981
Microwave; Caspases; Creatine kinase; Testosterone; Infertility
23.  Proteome profiling of embryo chick retina 
Proteome Science  2008;6:3.
Background
Little is known regarding the molecular pathways that underlie the process of retinal development. The purpose of this study was to identify proteins which may be involved in development of retina. We used a proteomics-based approach to identify proteins that are up- or down-regulated during the development of the embryo chick retina.
Results
Two-dimensional gel electrophoresis was performed with the retina of embryo chicken, which was obtained from embryos of day 7 (ED7) and of day 11 (ED11). The protein spots showing significant differences were selected for identification by MALDI mass spectrometry. Thirteen proteins were differentially expressed; seven proteins were up-regulated in embryo retina of chicken at ED 11 and six proteins were down-regulated. Significant proteins were also evaluated in embryo day 15 (ED15). Some of identified proteins were known to regulate cell proliferation, cell death, transport, metabolism, organization and extracellular matrix, and others also included novel proteins.
Conclusion
We identified thirteen proteins which differentially expressed in embryonal retina of chicken at day 7, as compared to the retina of embryo of day 11. They were various regulatory proteins for cellular signaling.
doi:10.1186/1477-5956-6-3
PMCID: PMC2267454  PMID: 18208622
24.  Infectious Bursal Disease in New Brunswick 
A flock of four week old chickens experienced a disease of sudden onset in which the only symptoms were those of depression shortly before death, and in which the predominant histological lesion was necrosis of lymphocytes in the bursa of Fabricius.
A virus, designated strain Sk-1, was isolated from pooled bursal tissue of affected birds and was serologically identified as a strain of the infectious bursal agent. This virus was chloroform resistant, did not hemagglutinate guinea pig or chicken erythrocytes and did not produce a cytopathic effect in chick embryo tissue cultures. Equivocal results were obtained in filtration studies but the agent was less than 100nm in diameter.
Four week old chicks inoculated with strain Sk-1 developed microscopic lesions in the bursa of Fabricius which were similar to those seen in the original field specimens. Inoculated chick embryos exhibited characteristic macroscopic lesions and necrosis of vascular tissue was a common histological change.
A limited serological survey of local poultry flocks indicated that infection by this agent had occurred in four of the ten flocks examined.
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PMCID: PMC1319791  PMID: 4356315
25.  Binge Toluene Exposure Alters Glutamate, Glutamine and GABA in the Adolescent Rat Brain as Measured by Proton Magnetic Resonance Spectroscopy* 
Drug and alcohol dependence  2010;115(1-2):101-106.
Despite the high incidence of toluene abuse in adolescents, little is known regarding the effect of binge exposure on neurochemical profiles during this developmental stage. In the current study, the effects of binge toluene exposure during adolescence on neurotransmitter levels were determined using high-resolution proton magnetic resonance spectroscopy ex vivo at 11.7 T. Adolescent male Sprague-Dawley rats were exposed to toluene (0, 8,000 , or 12,000 ppm) for 15 min twice daily from postnatal day 28 (P28) through P34 and then euthanized either one or seven days later (on P35 or P42) to assess glutamate, glutamine, and GABA levels in intact tissue punches from the medial prefrontal cortex (mPFC), anterior striatum and hippocampus. In the mPFC, toluene reduced glutamate one day after exposure, with no effect on GABA, while after seven days, glutamate was no longer affected but there was an increase in GABA levels. In the hippocampus, neither GABA nor glutamate was altered one day after exposure, whereas seven days after exposure, increases were observed in GABA and glutamate. Striatal glutamate and GABA levels measured after either one or seven days were not altered after toluene exposure. These findings show that one week of binge toluene inhalation selectively alters these neurotransmitters in the mPFC and hippocampus in adolescent rats, and that some of these effects endure at least one week after the exposure. The results suggest that age-dependent, differential neurochemical responses to toluene may contribute to the unique behavioral patterns associated with drug abuse among older children and young teens.
doi:10.1016/j.drugalcdep.2010.11.001
PMCID: PMC3071441  PMID: 21126832
Toluene; adolescents; glutamate; GABA; magnetic resonance spectroscopy; neuropharmacology; imaging

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