Feulgen Research Articles

Genome Size and Chromosome Number Evolution in Korean Iris L. Species (Iridaceae Juss.)

Chromosome numbers, karyotypes, and genome sizes of 14 Iris L. (Iridaceae Juss.) species in Korea and their closely related taxon, Sisyrinchium rosulatum, are presented and analyzed in a phylogenetic framework. To date, understanding the chromosomal evolution of Korean irises has been hampered by their high chromosome numbers. Here, we report analyses of chromosome numbers and karyotypes obtained via classic Feulgen staining and genome sizes measured using flow cytometry in Korean irises. More than a two-fold variation in chromosome numbers (2n = 22 to 2n = 50) and over a three-fold genome size variation (2.39 pg to 7.86 pg/1 C) suggest the putative polyploid and/or dysploid origin of some taxa. Our study demonstrates that the patterns of genome size variation and chromosome number changes in Korean irises do not correlate with the phylogenetic relationships and could have been affected by different evolutionary processes involving polyploidy or dysploidy. This study presents the first comprehensive chromosomal and genome size data for Korean Iris species. Further studies involving molecular cytogenetic and phylogenomic analyses are needed to interpret the mechanisms involved in the origin of chromosomal variation in the Iris.

In Memoriam: Marvin A. Van Dilla: 1919-2019.

© 2020 International Society for Advancement of Cytometry.

Development of the female gametophyte of Agave colimana, subgenus Littaea (Asparagaceae)1

Agave colimana, subgenus Littaea (Asparagaceae) is a wild plant that belongs to the Asparagaceae family, and in this species, the formation of its female reproductive system is unknown, probably because it has no commercial use. However, A. colimana recently has been considered as a candidate for marketing as a succulent plant because of its floral and growth architecture. Knowledge about aspects of the formation of the embryo sac is useful for understanding its sexual reproduction, which is important for propagation by seed and breeding purposes. Megasporogenesis and megagametogenesis were studied using Feulgen staining under confocal microscopy. Megasporogenesis resulted in the formation of a linear tetrad with the functional megaspore attached to the chalazal end. Subsequently, megagametogenesis from the functional megaspore was followed by three mitotic divisions generating an embryo sac with seven cells: two synergid cells, one egg cell, three antipodal cells, and one central cell with a 2n nucleus. The embryo sac of Agave colimana was found to be monosporic of the Polygonum-type considered as the typical type in the majority of angiosperm plants. Also, abnormal embryo sacs were observed, following possible errors during meiosis.

Demonstration of Deoxyribonucleic Acid in Colorectal Cancer Using Feulgen reaction

Demonstration of Deoxyribonucleic Acid in Colorectal Cancer Using Feulgen reaction

Integrin-Linked-Kinase Overexpression Is Implicated in Mechanisms of Genomic Instability in Human Colorectal Cancer.

Genomic instability is a hallmark of cancer cells contributing to tumor development and progression. Integrin-linked kinase (ILK) is a focal adhesion protein with well-established role in carcinogenesis. We have previously shown that ILK overexpression is critically implicated in human colorectal cancer (CRC) progression. In light of the recent findings that ILK regulates centrosomes and mitotic spindle formation, we aimed to determine its implication in mechanisms of genomic instability in human CRC. Association of ILK expression with markers of genomic instability (micronuclei formation, nucleus size, and intensity) was investigated in diploid human colon cancer cells HCT116 upon ectopic ILK overexpression, by immunofluorescence and in human CRC samples by Feulgen staining. We also evaluated the role of ILK in mitotic spindle formation, by immunofluorescence, in HCT116 cells upon inhibition and overexpression of ILK. Finally, we evaluated association of ILK overexpression with markers of DNA damage (p-H2AX, p-ATM/ATR) in human CRC tissue samples by immunohistochemistry and in ILK-overexpressing cells by immunofluorescence. We showed that ILK overexpression is associated with genomic instability markers in human colon cancer cells and tissues samples. Aberrant mitotic spindles were observed in cells treated with specific ILK inhibitor (QLT0267), while ILK-overexpressing cells failed to undergo nocodazole-induced mitotic arrest. ILK overexpression was also associated with markers of DNA damage in HCT116 cells and human CRC tissue samples. The above findings indicate that overexpression of ILK is implicated in mechanisms of genomic instability in CRC suggesting a novel role of this protein in cancer.

Use of DNA-specific stains as indicators of nuclei and extranuclear substances in leaf cells of the Middle Eocene Metasequoia from Arctic Canada

For this study, we used leaves of fossilized Metasequoia nathorstii from the Middle Eocene sediments of the Axel Heiberg Island of the Canadian Arctic Archipelago, and the leaves of modern M. glyptostroboides collected from the garden of the Komarov Botanical Institute RAS. Feulgen staining of the Metasequoia fossil leaves revealed the preservation of Feulgen-positive, heterogeneously stained nuclear structures in the cells, similar to those in the cells of the modern plant. The nuclei were brown-red-stained, and the chromocenters were stained more intensely, suggesting that the general morphology of the cell nucleus was preserved and the aldehyde groups of the deoxyribose that were available to react with the Feulgen's basic fuchsin were preserved in the nuclear chromatin of the fossil plants. The additional staining of the preparations with the alcian blue and Ehrlich's hematoxylin displays the cell walls and the adjacent intracellular material that is colored from light to dark blue. Staining the preparations with dA + dT-specific fluorochrome DAPI induced a weak fluorescence of the fossil cell nuclei, indicating the presence of some dsDNA molecules in the Metasequoia fossil leaves.

Revised tests and standards for Biological Stain Commission certification of alcian blue dyes

ABSTRACTAlcian blue dyes are copper phthalocyanines with a variety of cationic side chains; they are useful for staining carbohydrate polyanions while avoiding staining of nucleic acids. The properties of the original alcian blue and of similar dyes with published chemical structures are reviewed here. Variation among samples submitted to the Biological Stain Commission (BSC) for certification has led to the recognition of two types of commercially available alcian blue at this time. The designation “alcian blue 8G or equivalent” is reserved for dyes that resemble alcian blue 8GX manufactured in the 1960s (CI 74240; ingrain blue 1). These dyes react with alkali to form an insoluble pigment that cannot be re-dissolved in acid. The name “alcian blue variant” is for similar dyes that do not form insoluble pigments; an alkali-induced precipitate, if formed, re-dissolves with acidification. For certification by the BSC, both types of alcian blue must dissolve in 3% acetic acid to make a 1% solution (pH close to 2.5), which must provide selective coloration of intestinal mucus, cartilage and mast cells, but not of nuclei. After alcian blue staining and treatment with 0.03 M Na2CO3 or Li2CO3 to convert the bound dye to a pigment, the Feulgen stain for DNA is applied. Dyes to be certified as alcian blue 8G or the equivalent must resist extraction by the 5 M HCl used in the Feulgen reaction. Dyes to be certified as alcian blue variant are not required to be convertible to acid-insoluble pigments, but they must dissolve easily in water at pH 5.7 containing 0.5 M magnesium chloride and the dye must remain in solution for at least 24 h. A critical electrolyte concentration (CEC) staining test also is described; this must be passed for certification of an alcian blue variant. Successful CEC staining is also a desirable property of alcian blue 8G or equivalent, but not essential for certification of an otherwise satisfactory batch. The spectrophotometric criteria for alcian blue dyes also are revised; a wider range of absorption maximum (605–634 nm) is allowed. The dye powders used in published staining techniques with the original alcian blue 8G were 40–60% dye, but some modern alcian blue dyes have dye content as high as 90%. The BSC’s assay for dye content is not a criterion for certification, but it should influence the amount of dye to include in a staining solution.

Imaging of Embryo Sac and Early Seed Development in Maize after Feulgen Staining.

Compared with small model plants like Arabidopsis containing ovules with few cell layers, embryo sac and embryo development of model crop plants such as maize and other grasses are difficult to image. Multiple layers of tissue usually surround the deeply embedded embryo sac and developing embryo. Moreover, reliable cell biological marker lines labeling, for example, nuclei, plasma membrane, cell walls, or cells of a specific identity are often not available. The introduction of markers to study mutants is difficult and time-consuming and may require several generations of backcrosses. In this chapter, we therefore present an easy protocol to image maize ovaries and developing embryo sacs before and after fertilization allowing also high-throughput mutant analysis. The laborious embedding of samples and preparation of thin sections are omitted in this fixing-Feulgen staining-clearing (FFC) method. Optical sectioning through multiple layers of tissue is possible allowing 3D reconstructions of the whole embryo sac if necessary. The advantage of staining cell nuclei using the FFC method described here compared, for example, with DAPI staining is a wide range of Schiff's type reagents available for the Feulgen reaction. Depending on the reagent of choice, various conditions such as different excitation/emission filters or even white light can be applied for imaging. Moreover, in order to better visualize cell division, nuclei polarity as well as cell extent and integrity, periodic acid staining (PAS) of cell walls can be combined with Feulgen staining.

Localized Toxicity in Patients with Fixed Orthodontic Appliance:A Case Control Study

Fixed orthodontic appliances made of metal alloys are routinely used in dental practise to correct malocclusion. Oral environment favours the biodegradation of metal alloys. The discharged metal ions accumulate cumulatively on the adjacent tissues and results in toxic effects on cellular metabolism and DNA stability. The study was designed to evaluate localized genetic and cytokinetic toxicity due to fixed orthodontic appliance by measuring micronuclei frequency and other nuclear abnormalities in the buccal mucosal cells. Subjects with orthodontic appliance were selected as exposure cohort (n=20) and subjects without orthodontic appliance were selected as comparative cohort (n=20). Buccal smears were collected and DNA specific feulgen stain was used. Micronuclei, nuclear bud and binucleate cells frequency were recorded and compared between the study groups. We found a significant increase in the frequency of cells with micronuclei (p<0.001) and binucleate cells(p=0.002) in the exposure cohort compared to controls. The increase in the indicators of genotoxicity and cytokinetic toxicity in the exposed cohort compared to controls could be attributed to the exposure to the metal ions released from long term orthodontic appliance treatment. Increase in binucleate cells in subjects with fixed orthodontic appliance was demonstrated for the first time in our study. The cellular changes due to toxicity could be the predisposing factor for the occurrence of immune mediated oral mucosal lesions in orthodontic patients.

Evaluation of Micronucleus Frequencies in Exfoliated Buccal Cells of Potentially Malignant Disorder Subjects Using Feulgen Stain

Evaluation of Micronucleus Frequencies in Exfoliated Buccal Cells of Potentially Malignant Disorder Subjects Using Feulgen Stain

DAÑO GENÉTICO Y CITOTÓXICO PROVOCADO POR PLAGUICIDAS EN JORNALEROS QUE LABORAN EN INVERNADEROS EN ATLIXCO, PUEBLA, MÉXICO

The monitoring of genotoxic damage in laborers of greenhouses allows the identification of deterioration of DNA originated by working conditions. It has been suggested the use of different biological markers to evaluate the risk of occupational exposure and DNA damage, one of them is the micronucleus (MN) assay. The objective of the study was to determine the frequency of MN and nuclear abnormalities in the buccal mucosa epithelium of 40 workers of high and low technology greenhouses, who are exposed to diferent kinds of pesticides, and of 40 people not exposed to these compounds, both populations from Atlixco City, in Puebla, Mexico. The samples taken were air-dried and fixed with methanol-acetic acid. Later they were stained by the Feulgen reaction. Nuclear abnormalities were assessed by counting one thousand differentiated basal cells per participant. Comparison of biomarkers frequencies indicates higher values of nuclear abnormalities statistically significant in laborers of greenhouses, whose frequency index ratios were: 63.0 binucleate cells, 14.2 cells with condensed chromatin, 8.0 cariolitic cells, 3.8 cells pycnotic, 2.4 cells with nuclear outbreaks, and 2.3 cells with micronuclei. The highest frequency of nuclear abnormalities was determined in laborers working in low-tech greenhouses compared to high-tech greenhouses. The male population of low-tech greenhouses showed higher frequencies of pycnotic cells, micronuclei, and cells with nuclear outbreaks.

Karyotype and genome size variation in Ajuga L. (Ajugoideae–Lamiaceae)

Chromosome number changes and karyotype evolution play an important role in plant genome diversification and eventually in speciation. The genus Ajuga L. (Lamiaceae) has approximately 50 species distributed in temperate to subtropical regions. Four of these species are currently recognized in Korea (A. decumbens Thunb., A. multiflora Bunge, A. nipponensis Makino and A. spectabilis Nakai). Understanding the karyotype evolution in Ajuga has been hampered by the small size of their chromosomes and symmetrical karyotypes. Here we used classic Feulgen staining to establish chromosome numbers and construct karyotypes of the four species of Ajuga recognized in Korea and flow cytometry was used to study their variation in genome. The chromosome number of all investigated plants was 2n = 32. Still, the 2C DNA content ranged from 2.18 pg (A. decumbens) to 4.53 pg (A. multiflora). While the chromosome numbers were the same for all investigated species, the genome size variation could potentially be used as a taxonomic marker.

MiR-34a affects hepatocyte proliferation during hepatocyte regeneration through regulating Notch/HIF-1α signaling pathway.

To explore the influences of micro ribonucleic acid (miR)-34a on liver function and hepatocyte proliferation during hepatocyte regeneration in rats and its mechanism. A total of 80 Sprague-Dawley rats were randomly divided into 4 groups: Sham-2 d group (2 days after hepatectomy), Sham-10 d group (10 days after hepatectomy), miR-34a siRNA-2d group (miR-34a knockdown + 2 days after hepatectomy) and miR-34a siRNA-10 d group (miR-34a knockdown + 10 days after hepatectomy), with 20 rats in each group. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) were detected at 2 d and 10 d after the operation. The rat liver was harvested for calculating the liver/body weight ratio. In addition, the deoxyribonucleic acid (DNA) content in rat hepatocytes was detected via Feulgen staining. The pathological changes in rat liver were detected via hematoxylin-eosin (H&E) staining. Moreover, the hepatocyte apoptosis in each group was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Expression levels of proliferating cell nuclear antigen (PCNA), Notch1 intracellular domain (NICD), and hypoxia-inducible factor-1α (HIF-1α) in liver tissues of each group were detected via immunohistochemistry and Western blotting. No significant differences in the liver/body weight ratio, serum levels of ALT, AST, LDH, pathological structure of the liver, hepatocyte apoptosis level, and PCNA expression in hepatocytes were found between miR-34a siRNA-2 d group and Sham-2 d group. However, the expression levels of NICD and HIF-1α in the liver significantly increased in miR-34a siRNA-2 d group compared with those in Sham-2 d group (p<0.05). On the contrary, compared with those in Sham-10 d group, the liver function and hepatocyte regeneration level significantly increased in miR-34a siRNA-10 d group. Increased liver/body weight ratio, remarkable decline in serum levels of ALT, AST, and LDH, significant alleviation of pathological injury of liver tissues, decreased the apoptosis level and upregulated PCNA protein were observed in miR-34a siRNA-10 d group than those of Sham-10 d group. The Notch/HIF-1α signaling pathway was also significantly activated. MiR-34a knockdown can significantly enhance the liver function and hepatocyte regeneration ability in rats at 10 d after hepatectomy through activating the Notch/HIF-1α signaling pathway.

Death of female flower microsporocytes progresses independently of meiosis-like process and can be accelerated by specific transcripts in Asparagus officinalis

Asparagus officinalis (garden asparagus) is a dioecious perennial crop, and the dioecy (i.e., sex) of A. officinalis can affect its productivity. In A. officinalis, flower anthers in female plants fail to accumulate callose around microsporocytes, fail to complete meiosis, and degenerate due to cell death. Although 13 genes have been implicated in the anther development of male and female flowers, it is unclear how these genes regulate the cell death in female flower anthers. The aim of this study was to narrow down factors involved in this process. TUNEL staining and Feulgen staining of female flower microsporocytes suggest that female microsporocytes enter a previously undetected meiosis-like process, and that the cell death occurs independently of this meiosis-like process, excluding the possibility that the cell death is caused by the cessation of meiosis. RNA sequencing with individual floral organs (tepals, pistils and stamens) revealed that several genes possibly regulating the cell death, such as metacaspase genes and a Bax inhibitor-1 gene, are differentially regulated between female and male flower anthers, and that genes involved in callose accumulation are up-regulated only in male flower anthers. These genes are likely involved in regulating the cell death in female flower anthers in A. officinalis.

DNA cytometry of exfoliated cells in the diagnosis of oral potential malignant disorders

To evaluate the diagnostic efficiency of oral mucosa disease, especially oral squamous cell carcinoma (OSCC) and oral potential malignant disorders (OPMDs) by DNA cytometry compared with histopathological diagnosis, so as to find a convenient, simple and low-invasive method for screening and follow-up. 203 subjects with OSCC, OPMDs and other oral mucosa disease without dysplasia according to the inclusion criteria and exclusion criteria were recruited from Peking University School and Hospital of Stomatology. The mean age was (52.44±13.55) years, 98 males and 105 females. Brush biopsy was taken before scalpel biopsy at the same site. The brush biopsy sample was screened by moticytometer system for DNA cytometry after Feulgen stain, and histopathological examination were taken for the scalpel tissue. Data from DNA cytometry were used to calculate the parameters, such as sensitivity, specificity, positive and negative predictive values, odds ratios, Youden index (YI), positive and negative likelihood ratios, compared with the golden standard, histopathological diagnosis. DNA cytometry and histopathological diagnosis were performed back to back. Totally, 42 OSCC and 4 tumor in situ (TIS), 39 oral leukoplakia (OLK) with dysplasia (17 mild dysplasia, 13 medium dysplasia and 9 severe dysplasia), 29 OLK with hyperplasia, 1 verrucous OLK, 83 oral lichen planus (OLP) and 5 inflammation were included in our research. We grouped the OSCC, TIS and dysplasia as the positive group and others without dysplasia as the negative group, the sensitivity of DNA cytometry was 79.07%, the specificity was 81.20%, and the diagnostic accuracy was 80.30%,We grouped the OSCC and TIS as the tumor group, OLP, OLK with hyperplasia and inflammation as the non-tumor group, The sensitivity of DNA cytometry in diagnosing OSCC and TIS was 95.65%, and the specificity was 81.2%, The diagnostic accuracy was 85.28%. positive predictive values 66.67%, negative predictive values 97.94%, ratio odds 95, positive likelihood ratio 5.09, negative likelihood ratio 0.05, and Youden index 0.77. For the dysplasia, we grouped the different dysplasia together as the dyaplasia group, OLP, OLK with hyperplasia and inflammation as the non-tumor group, the sensitivity of DNA cytometry in diagnosing dyaplasia is 60%, the specificity is 81.2%. The diagnostic accuracy is 75.8%, positive predictive values 52.17%, negative predictive values 85.59%, ratio odds 6.48, positive likelihood ratio 3.19, negative likelihood ratio 0.49, and Youden index 0.41. DNA cytometry is convenient and low-invasive, which can be used as an adjuvant method for screening the early OSCC and OPMDs, monitoring the prognosis of OSCC after surgery. Further large-scale and long period prospective studies are necessary to validate the better value of DNA cytometry.

Mitotic figures evaluation in oral squamous cell carcinoma using crystal violet and feulgen stains - A comparative study

Context: Mitosis is the process of nuclear cell division. Increased mitosis causes excessive proliferation of cells which is observed in oral squamous cell carcinoma (OSCC). However, there is always problem in differentiating a mitotic cell from an apoptotic cell, during routine staining procedures, which affect the reliability of histological grading of OSCC. Recently, several methods have been implemented in identifying these mitotic figures (MFs), but they seem to be time-consuming and expensive which makes them less feasible. Thus, an effort was done to evaluate the efficacy of crystal violet and Feulgen stains in identifying MFs along with routine hematoxylin and eosin (H and E) in OSCC. Aim: In the identification of MFs in diagnosed cases of OSCC using crystal violet and Feulgen stains and H and E stain. Settings and Design: This was a retrospective study. Materials and Methods: The study sample includes five tissue sections of moderately-differentiated and well-differentiated OSCC each and four sections of poorly differentiated which were stained with H and E, Feulgen, and 1% crystal violet stains, and the number of MFs was enumerated. Statistical Analysis Used: One-way ANOVA was used for statistical analysis. Results: The results from our study showed that 1% crystal violet-stained MFs are better than H and E and Feulgen stains. Conclusion: Crystal violet stain can be considered as a simple, reliable, cost-effective, and reproducible method of staining MFs.

Feulgen Reaction as a Marker for Identifying Prostatic Hyperplasia and Prostate Cancer

Introduction: A prostate cancer case in sub-Saharan Africa is rising. In Sudan, prostate cancer ranks third at after breast cancer. The nuclear deoxyribonucleic acid content has been shown to be of great prognostic value in prostate tumors. In the present work, we focus on semi-quantification of nuclear deoxyribonucleic acid in prostate cancer compared to prostatic hyperplasia using the Feulgen reaction technique. Methods: Paraffin wax sections were selected from patients previously diagnosed with prostatic hyperplasia and prostatic cancer. Clinic pathological data of patients were collected from the records of the archive of the National Health Laboratory Khartoum Sudan. From all cases, two sections were taken. One section was stained with Hematoxylin and eosin stain to confirm the diagnosis, while the other paraffin section was used for deoxyribonucleic acid demonstration using Feulgen reaction. Results: A total of 46 patients with a clinical and pathologic diagnosis of prostatic tumors were enrolled in this study; 23 (50.1%) had high-grade prostatic adenocarcinoma, 11 (23.9%) had moderate grade prostatic adenocarcinoma, two (4.3%) had low-grade prostatic adenocarcinoma and 10 (21.7%) had benign hyperplasia. The significant differences in deoxyribonucleic acid staining intensities were observed in high-grade prostatic adenocarcinomas. The deoxyribonucleic acid staining intensities of moderate and high-grade prostatic adenocarcinomas were significantly higher than those of benign hyperplasia (P<0.000). Conclusion: Deoxyribonucleic acid detection using Feulgen reaction may provide valuable prognostic information in prostate cancer, with potential clinical implications in patient management, active research on this topic should be a high priority objective.

Utility of DNA-Specific Stains in Micronuclei Assay as a Marker of Genotoxicity in Oral Potentially Malignant Disorders and Oral Squamous Cell Carcinoma.

Background:The term oral potentially malignant disorder (OPMD) was recommended to refer to precancer as it conveys that not all disorders described under this term may transform into cancer. Oral squamous cell carcinoma (OSCC) arises through an accumulation of genetic alterations, deoxy ribonucleic acid (DNA) changes, and epigenetic alterations. Thus, a simple yet a sensitive and specific test for early diagnosis is the need of an hour. The micronuclei (MN) assay in exfoliated epithelial cells is potentially an excellent biomarker to detect chromosome loss or malfunction of mitotic spindle.Aim of the Study:To compare the frequency of MN in exfoliated cells from oral mucosa exposed to genotoxic agents using different staining procedures and to observe the incidence of micronucleus in potentially malignant and malignant lesions.Materials and Methods:The study was undertaken to observe the cytogenetic damage in the exfoliated buccal cells of 75 cases of tobacco-related PMDs, OSCC and control subjects (25 cases from each group) and were evaluated with nonspecific May-Grünwald Giemsa stain and DNA-specific Feulgen stain. The results were statistically determined using SPSS version 17.0.Results:Correlation analyses in the present study depicted that MN frequency was significantly more in oral squamous cell carcinoma than OPMDs and normal group (P < 0.05). Giemsa-stained slides correlated significantly with karyorrhexis, karyolysis, condensed chromatin, and binucleates, whereas no such correlations were found with DNA-specific stains.Conclusion:Malignant transformation is accompanied by loss of cell capacity to evolve to death in situations of DNA damage. These findings indicate that nuclear anomalies may be misinterpreted as MN with nonspecific DNA stains and lead to false-positive results in studies with cells of epithelial origin.

Suitability of Papanicolaou stain for micronuclei mass screening in buccal smears comparing reference feulgen stain and papanicolaou stain in field

&lt;strong&gt;Background:&lt;/strong&gt; Micronuclei (MN) assessment so far was done directly, individually, and intra- and, here, inter-observer bias could not be eliminated. It was subjective. &lt;strong&gt;Methods:&lt;/strong&gt; In this study, accurate and reproducible MN counting was facilitated by the use of liquid cytology and image capturing with MN identification analysis. &lt;strong&gt;Results:&lt;/strong&gt; Comparing the two nuclear staining techniques - DNA specific Feulgen stain and commonly available Papanicolaou (PAP) stain we found that both techniques gave statistically comparable accurate and reproducible MN results. T-test and non-parametric followed by paired non-parametric and then Wilcoxon matched-pairs signed rank test tell us that both the staining procedures have no difference at P &amp;lt;0.05. &lt;strong&gt;Conclusion:&lt;/strong&gt; These protocols are comparable and reproducible, and may be adoptable for mass screening to assess the genotoxic effects of substances like tobacco.

Effect of monosodium glutamate on liver of pregnant rats and their fetuses (Histological and histochemical studies)

Background: monosodium glutamate (MSG) is a widely used flavor enhancer and stabilizer in readymade or packaged foods. The excessive use of MSG increases the oxidative stress in the different body systems and causes teratogenic effects on fetuses of mothers treated with it in the gestational periods. Aim of the work: this study investigated effects of MSG on maternal and fetal liver tissue. Material and Methods: the present work was carried out on fifty mature pregnant Albino rats (Weight 200 ± 20 g) and six male rats in the reproductive age (Older than 60 days). The first group was served as the control group. The remnant pregnant rats were categorized into two sub- groups. Animals of the 1st sub group were dissected on day 15th of gestation (C15) and the second was dissected on day 19th (C19) of gestation. The second group was administrated monosodium glutamate daily with the dose of 7 g/10mL/kg body weight from the 9th to 14th of gestation. Then the pregnant mothers were categorized into two sub- groups, the first was dissected on the 15th day (G15) and the second on the 19th day (G19) of gestation. The pregnant mothers were dissected after anesthesia which was performed by Halothane produced by Pharco. Comp. The uterine horns were removed from the surrounding fats then fetuses were removed from the surrounding amniotic membranes. Histopathological and histochemical studies were done on the maternal and fetal liver tissue through hematoxylin-eosin stain and Feulgen reaction. Results: numerous degenerative changes were detected in both maternal and fetal liver tissues of G15 and G19 groups. These changes included: vacuolation in hepatocytes, degenerative and necrotic areas and atrophied size of hepatocytes with pyknotic nuclei. These changes were observed in the centrolobular zones and the portal zones. Conclusions: This study showed that short-term MSG exposure in the gestational periods caused several histological and histochemical changes in the maternal and fetal liver tissues. These findings should be re-evaluated in additional long-term studies.