scholarly journals Affinity binding of non-histone chromatin proteins to the X chromosome of Drosophila by in situ chromatin reconstitution and its significance

1986 ◽  
Vol 86 (1) ◽  
pp. 35-45
Author(s):  
M. Ukil ◽  
K. Chatterjee ◽  
A. Dey ◽  
S. Ghosh ◽  
A.S. Mukherjee
Keyword(s):  
Binding Affinity ◽  
X Chromosome ◽  
Positive Control ◽  
Negative Control ◽  
Chromosomal Protein ◽  
X Chromosomes ◽  
Male And Female ◽  
Interference Band ◽  
Band Filter

Cytophotometric analysis of the in situ binding affinity of non-histone chromosomal protein (NHCP) to the polytenic X chromosome and autosome of Drosophila melanogaster has been carried out using Feulgen-Napthol Yellow S staining technique. The results reveal that the mean transformed absorbance ratio (male:female) with a 547 nm interference band filter for the two specific segments of the X chromosome is close to 0.5, while for a specific segment of an autosome it is close to 1.0, in the two sets of control; namely, the positive control (no treatment) and the negative control (treated with 1 M-urea+2M-NaCl) as well as in the reconstituted chromosomal preparations, which received 1 M-urea+2M-NaCl and the NHCP isolated from D. melanogaster. In contrast, the transformed absorbance ratios (male:female) with a 433 nm interference band filter yielded an interestingly different result. The ratios with a 433 nm filter for the X chromosome segments are significantly greater than 0.5 in all three sets of experiments. This finding by itself suggests that the NHCP binding affinity is dissimilar for the X chromosomes of male and female. When the 433 to 547 nm absorbance ratios were compared among the three sets, the data clearly revealed that in both positive control and NHCP reconstituted samples, the absorbance ratios (i.e. 433:547 nm) are significantly different between X chromosomes from males and those from females, while they are different between autosomes from males and females. The ratios are also not significantly different between male and female, either for the X chromosome or for the autosome in the negative control. These findings, therefore, suggest that there is a stronger binding affinity of NHCP for the male X chromosome of Drosophila, and reinstate the view that the X-chromosomal hyperactivity in male Drosophila is the consequence of a regulated organizational change in the DNA template.

A Novel DNA/RNA FISH X Inactivation Assay Reveals a Nonrandom, Ploidy-Dependent Acquisition of the Active and Inactive X Chromosomes in Childhood Hyperdiploid Acute Lymphoblastic Leukemia (ALL) and Non-Hodgkin Lymphoma (NHL).

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1080-1080
Author(s):  
Oskar A. Haas ◽  
Petra Zeitlhofer ◽  
Sabine Strehl ◽  
Michael Pfeilstoecker ◽  
Margit Koenig ◽  
...  
Keyword(s):  
Acetic Acid ◽  
X Chromosome ◽  
Dna Sequences ◽  
X Chromosomes ◽  
Childhood All ◽  
Male And Female ◽  
Female Patients ◽  
Rna Fish ◽  
Random Fashion ◽  
Male Patients

Abstract The most common numerical chromosome aberration in childhood ALL and NHL is the gain of an extra X chromosome in both male and female patients. We were therefore interested to investigate whether this non-disjunction event affects the active and inactive X chromosomes in a random or non-random fashion. In female cases both the active or inactive X may be duplicated randomly or non-randomly, whereas in male patients only the solitary active homologue can be copied. However, in theory a duplicated active X might subsequently also be subjected to de novo inactivation in both sexes. The inactivation status of acquired X chromosomes is usually evaluated by methylation-specific PCR (MS-PCR), which allows the simultaneous quantification of various differentially methylated polymorphic DNA sequences on the X chromosome, such as those contained in the HUMARA or FMR1 genes. Previous evidence from such analyses suggested that in NHL patients the acquired X chromosomes are and remain always active in male patients, whereas in females both the active and inactive X are duplicated in a random fashion (McDonald et al, Genes, Chromosomes & Cancer 28:246;2000). In childhood ALL this issue has not yet been investigated. However, quantification with MS-PCR has its limitations, especially in cases with low blast cells numbers. To overcome this problem, we have therefore developed a simultaneous dual-color DNA/RNA FISH assay that enables the enumeration of active and inactive X chromosomes on a single cell level. FISH was performed with probes specific for the X centromere and the XIST RNA, which is exclusively expressed from and covers vast parts of the inactive X in human interphase cells. Following the successful evaluation of the assay on methanol/acetic acid-fixed cells that were obtained from 10 healthy individuals and 23 cases with various constitutional X chromosome aneuploidies, we analyzed 54 methanol/acetic acid-fixed samples from hyperdiploid cases of childhood ALL and 29 from NHL. The ALL cases comprised 24 males with two X, 23 females with three X and seven females with four X. The NHL cases consisted of 18 male (9 in the hypo- to hyperdiploid and 9 in the pseudotriploid to pseudotetraploid range) and 11 female patients (7 with three X and 4 with four X chromosomes). In contrast to all constitutional control samples, which as expected contained only one active X, two of the three X in leukemic cell samples from both male and female patients were active. The only exception was a male patient, who most likely was a Klinefelter syndrome with a constitutional XXY. In contrast, all female patients with four X had duplicated both the active and inactive X chromosome. These findings prove that irrespective of the sex of the patient, the active X is exclusively duplicated in cases with three X chromosomes. The consistent gain of both the active and inactive X in female cases with four X, on the other hand, further corroborates previously established evidence that in all instances a single non-discjunction event leads to the maldistribution of chromosomes irrespective of the ploidy range. Moreover, the exclusive presence of duplicated active X chromosomes in hyperdiploid ALL concurs with and explains the results of gene expression profiling studies, which have shown a corresponding over-expression of X-encoded genes.

Involvement of the X chromosome in fertility of male and female hybrids of Glossina morsitans morsitans and Glossina morsitans centralis (Diptera: Glossinidae)

1989 ◽  
Vol 67 (4) ◽  
pp. 869-871 ◽  
Author(s):  
R. H. Gooding
Keyword(s):  
X Chromosome ◽  
Glossina Morsitans Morsitans ◽  
The Other ◽  
Marker Genes ◽  
Glossina Morsitans ◽  
X Chromosomes ◽  
Male And Female ◽  
Intrachromosomal Recombination ◽  
Glossina Morsitans Centralis ◽  
Glucose 6 Phosphate Dehydrogenase

In hybrid females of Glossina morsitans morsitans Westwood and Glossina morsitans centralis Machado that carried four well-separated marker genes, suppression of intrachromosomal recombination occurred between the loci for glucose-6-phosphate dehydrogenase (G6pd) and arginine phosphokinase (Apk) on the X chromosome. Fertility of backcross females was not influenced by whether they mated with G. m. morsitans or G. m. centralis, but it was higher in females that received both of their X chromosomes from G. m. morsitans than it was in females that received one X chromosome from G. m. morsitans and the other from G. m. centralis.

scholarly journals The morfological and molecular identification of Fusarium verticillioides causing fusariosis on wheat grain

Genetika ◽  
2021 ◽  
Vol 53 (2) ◽  
pp. 641-649
Author(s):  
Iva Savic ◽  
Milica Nikolic ◽  
Ana Nikolic ◽  
Ivana Vico ◽  
Snezana Mladenovic-Drinic ◽  
...  
Keyword(s):  
Fusarium Head Blight ◽  
Fusarium Verticillioides ◽  
Positive Control ◽  
Negative Control ◽  
Potato Dextrose Agar ◽  
Wheat Grain ◽  
Head Blight ◽  
Reference Isolate ◽  
Wheat Leaves

During the 2014-2017 period, wheat samples were collected from discoloured spikes affected by Fusarium head blight (FHB) from 20 locations in Serbia. After isolation, fungi were cultivated on potato dextrose agar (PDA) at 25oC for 7 days. Based on the in situ identification on PDA, 36 isolates of the section Liseola were selected for further analyses. The pathogenicity of all isolates was confirmed on wheat leaves. The virulence of isolates was determined by measuring the lengths of spots formed at the inoculation leaf site. In order to prove the presence of the species Fusarium verticillioides, a pair of primers FV-F2/FV-R was used. This pair of primers amplifies the sequence of the gaoB gene, and it proved to be specific for the stated species. Moreover, for the same purpose, a pair of primers VER1-VER2 based on the calmodulin partial gene was used. The reference isolate RBG 1603 Q27 was used as a positive control. The pair of primers VER1-VER2 produced a band of the expected size - 578 bp in 18 isolates, while using FV-F2/FV-R, a 370bp long band confirmed the presence of F. verticillioides in 16 samples. Sixteen out of 18 isolates verified with VER1-VER2 were also identified as F. verticillioides with FV-FS/FV-R. No amplification was observed in a negative control.

scholarly journals UJI EFEK ANALGESIK EKSTRAK DAUN MAHKOTA DEWA (Phaleria macrocarpa) PADA MENCIT (Mus musculus)

2014 ◽  
Vol 1 (2) ◽  
Author(s):  
Dinar Salsabila Tone ◽  
Christi Mambo
Keyword(s):  
Treatment Group ◽  
Medicinal Plant ◽  
Mus Musculus ◽  
Analgesic Effect ◽  
Positive Control ◽  
Negative Control ◽  
Control Group ◽  
Male And Female ◽  
Average Value ◽  
Phaleria Macrocarpa

ABSTRACTThe plant of Mahkota Dewa is a traditional plant which is used as a medicinal plant whose benefits are located in almost parts where it contains flavonoid and saponin compounds that have a variety of effects and one of them is analgesic effect. This research aims to determine the analgesic effect of the extract of Mahkota Dewa leaf (Phaleria macrocarpa) in mices (Mus musculus). This research uses an experimental method using nine male and female mices which are divided into three groups: the positive control group that was given aspirin and the negative control that was given aquades and the treatment group that was given the extract of the Mahkota Dewa leaf. The research is done by giving the stimulus of pain in the form of heat 55o.C and then observes the response of the tested animal such as jumping or licking its legs and at the minute of 0 before treatment, and at the minutes of 30, 60, 90, 120 after the treatment. The average value of the number of respons of mices which were given the extract of the Mahkota Dewa leaf decreases from the 30th minute until the 90th minute. Conclusion. The extract of Mahkota Dewa leaf has an analgesic effect in Mouse.Key Word: Analgesic, Aspirin, Mahkota Dewa leaf (Phaleria macrocarpa)ABSTRAKTanaman mahkota dewa merupakan tumbuhan tradisional yang digunakan sebagai tumbuhan obat yang manfaatnya terletak hampir di seluruh bagian dimana di dalamnya terkandung senyawa-senyawa flavonoid dan saponin yang mempunyai bermacam-macam efek dan salah satunya adalah efek analgesik. Penelitian ini bertujuan untuk mengetahui efek analgesik dari ekstrak daun mahkota dewa (Phaleria macrocarpa) pada mencit (Mus musculus). Penelitian ini menggunakan metode eksperimental dengan menggunakan 9 ekor mencit jantan dan betina yang dibagi atas 3 kelompok yaitu kelompok kontrol positif yang diberi obat aspirin, kontrol negatif yang diberi aquades dan kelompok perlakuan yang diberi ekstrak daun mahkota dewa. Penelitian dilakukan dengan cara memberi rangsangan nyeri berupa suhu panas 55o.C kemudian mengamati respon hewan uji berupa melompat dan atau menjilat kaki pada menit ke-0 sebelum perlakuan, dan pada menit ke-30, 60, 90, 120 setelah perlakuan. Nilai rata-rata jumlah respon mencit yang diberikan ekstrak daun mahkota dewa mengalami penurunan dari menit ke-30 sampai menit ke-90. Kesimpulan. Ekstrak daun mahkota dewa memiliki efek analgesik pada mencit.Kata kunci: Analgesik, Aspirin, Daun Mahkota Dewa (Phaleria macrocarpa)

scholarly journals Antioxidants Effectivity In Skin Lotion Formulation Of Mesocarp Fruit Extract Lontar (Borassus Flabellifer) Against White Rats Wistar Male In-Situ

2017 ◽  
Vol 2 (01) ◽  
pp. 25
Author(s):  
Luthfiasari Amatullah ◽  
Tri Nur Cahyaningrum ◽  
Anisa Nur Fidyaningsih
Keyword(s):  
Positive Control ◽  
Negative Control ◽  
Skin Preparation ◽  
Palm Fruit ◽  
White Rats ◽  
Group 2 ◽  
The Stability ◽  
Palm Fruits ◽  
Control Compound

<p>Study of bioactive mesocarp on palm fruits has been conducted previously. Therefore, this research aims to make a preparation of antioxidant skin lotion palm fruit mesocarp (<em>Borrasus flabellifer</em>) and to evaluate the effects of antioxidants on skin preparation lotion palm fruit (<em>Borrasus flabellifer</em>) by In-situ.</p><p>This study was an experimental study by making a preparation of skin lotion on the stability of the corresponding and testing the antioxidant treatment with a sample of 30 rats male wistar strain. The antioxidant test was divided into 5 groups randomly: negative control (-), positive control (compound sunscreen brands parasol SPF 33), group 1 (lotio extract palm fruit mesocarp with a concentration of 0.2%), group 2 (lotio palm fruit mesocarp extract with concentration of 0.4%) and group 3 (lotio palm fruit mesocarp extract with concentration 0.8%).</p><p>The results of in-situ states showed 0.8% extract of palm fruit mesocarp exhibited the least erythema score which was an average of 1.7. It was comparable to positive control which exhibited similar erythema score.</p>

scholarly journals Effect of Milk Renewal on Cell Viability In Vitro at Different Time Frames

2017 ◽  
Vol 28 (4) ◽  
pp. 435-439 ◽  
Author(s):  
Beatriz Dulcineia Mendes de Souza ◽  
Ana Maria Hecke Alves ◽  
Dayane Machado Ribeiro ◽  
Luciane Geanini Pena dos Santos ◽  
Claudia Maria de Oliveira Simões ◽  
...  
Keyword(s):  
Cell Viability ◽  
Tap Water ◽  
Positive Control ◽  
Negative Control ◽  
Tetrazolium Salt ◽  
Periodontal Ligament Fibroblasts ◽  
Skimmed Milk ◽  
Time Frames

Abstract The purpose of this study was to evaluate if the renewal of milk as a storage medium, every 12, 24 and 48 h, is able to increase its ability to maintain human periodontal ligament fibroblasts (PDLF) viability over time. PDLF were soaked in Minimum Essential Medium at 37 °C (MEM-37) (positive control), tap water (Water) (negative control) and in skimmed milk (44 wells) at 5 °C and 20 °C. The skimmed milk was renewed every 12 h (Milk-12), 24 h (Milk-24) and 48 h (Milk-48) in 11 wells of each plate, and the milk in the remaining 11 wells of each plate was maintained in situ (not renewed milk) (NRM). After 24, 48, 72, 96 and 120 h, cell viability was determined by the tetrazolium salt-based colorimetric (MTT) assay. Data were statistically analyzed by Kruskal-Wallis, Scheffé and Mann-Whitney tests (a=5%). At 5 °C, only Milk-48 was significantly better than NRM. At 20 °C, NRM was more effective than Milk-12 and Milk-24 in all time periods. In relation to the temperature (5 °C or 20 °C), renewal of milk at 5 °C was better in maintaining cell viability than the renewal at 20 °C. In conclusion, the renewal of milk was able to increase its ability to maintain cell viability only when performed every 48 h in milk maintained at 5 °C.

Effects of dietary stachyose levels on caecal skatole concentration, hepatic cytochrome P450 mRNA expressions and enzymatic activities in broilers

2020 ◽  
Vol 124 (10) ◽  
pp. 1013-1020
Author(s):  
Hai-Ying Liu ◽  
Xin-Yun Zhao ◽  
Gui-Qin Yang ◽  
Ji-Zhe Liu ◽  
Xin Zhu
Keyword(s):  
Cytochrome P450 ◽  
Ph Value ◽  
Positive Control ◽  
Negative Control ◽  
Enzymatic Activities ◽  
Control Groups ◽  
Male And Female ◽  
Significant Difference ◽  
Soyabean Meal ◽  
Hepatic Cytochrome

AbstractEffects of dietary supplemental stachyose on caecal skatole concentration, hepatic cytochrome P450 (CYP450, CYP) mRNA expressions and enzymatic activities in broilers were evaluated. Arbor Acre commercial mixed male and female chicks were assigned randomly into six treatments. The positive control (PC) diet was based on maize–soyabean meal, and the negative control (NC) diet was based on maize–non-soyabean meal. The NC diet was then supplemented with 4, 5, 6 and 7 g/kg stachyose to create experimental diets, named S-4, S-5, S-6 and S-7, respectively. Each diet was fed to six replicates of ten birds from days 1 to 49. On day 49, the caecal skatole concentrations in the PC, S-4, S-5, S-6 and S-7 groups were lower than those in the NC group by 42·28, 23·68, 46·09, 15·31 and 45·14 % (P < 0·01), respectively. The lowest pH value was observed in the S-5 group (P < 0·05). The stachyose-fed groups of broilers had higher caecal acetate and propionate levels compared with control groups, and propionate levels in the S-6 and S-7 groups were higher than those in the S-4 and S-5 groups (P < 0·001). The highest CYP3A4 expression was found in the S-7 group (P < 0·05), but this was not different from PC, S-4, S-5 and S-6 treatments. There was no significant difference in CYP450 (1A2, 2D6 and 3A4) enzymatic activities among the groups (P > 0·05). In conclusion, caecal skatole levels can be influenced by dietary stachyose levels, and 5 g/kg of stachyose in the diet was suggested.

scholarly journals Efficacy of 0.05% Chlorhexidine and 0.05% Cetylpyridinium Chloride Mouthwash to Eliminate Living Bacteria on In Situ Collected Biofilms: An In Vitro Study

Antibiotics ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 730
Author(s):  
Kathrin Becker ◽  
Giulia Brunello ◽  
Luisa Scotti ◽  
Dieter Drescher ◽  
Gordon John
Keyword(s):  
Side Effects ◽  
Cetylpyridinium Chloride ◽  
Antibacterial Properties ◽  
In Vitro Study ◽  
Positive Control ◽  
Negative Control ◽  
Sterile Saline ◽  
Viability Assay

Chlorhexidine (CHX) mouthwashes are frequently used as an adjunctive measure for the treatment of periodontitis and peri-implantitis, as well as in patients on maintenance therapy. However, their prolonged use is associated with several side effects. This study aimed at evaluating if a mouthwash with a reduced concentration of CHX combined with cetylpyridnium chloride (CPC) was as effective as a conventional CHX mouthwash in the reduction in living cells in oral biofilms attached to hydroxyapatite (HA) and micro-rough titanium (Ti) surfaces. Four healthy volunteers wore a customized acrylic appliance containing HA and Ti discs for in situ plaque accumulation. Biofilms were grown on the discs for 24 or 48 h and then randomly exposed for 60 s to: 0.05% CHX + 0.05% CPC, 0.1% CHX (positive control) or sterile saline (negative control). Viability assay and live-dead staining were performed to quantify bacterial viability and to distinguish live and dead cells, respectively. At both time points, contrary to saline, CHX, both alone and in combination with CPC, exhibited high antibacterial properties and induced a significant reduction in biofilm viability. This study demonstrates the potential of mouthwashes containing a low concentration of CHX combined with CPC as effective antibacterial agents for long-term applications with reduced undesired side effects.

scholarly journals Divergent DNA methylation signatures of X chromosome regulation in marsupials and eutherians

2020 ◽  
Author(s):  
Devika Singh ◽  
Dan Sun ◽  
Andrew G. King ◽  
David E. Alquezar-Planas ◽  
Rebecca N. Johnson ◽  
...  
Keyword(s):  
Dna Methylation ◽  
X Chromosome ◽  
Specific Expression ◽  
X Chromosomes ◽  
Phascolarctos Cinereus ◽  
Male And Female ◽  
Inactive X Chromosome ◽  
Genome Wide ◽  
Promoter Dna Methylation ◽  
Promoter Dna

AbstractX chromosome inactivation (XCI) mediated by differential DNA methylation between sexes is well characterized in eutherian mammals. Although XCI is shared between eutherians and marsupials, the role of DNA methylation in marsupial XCI remains contested. Here we examine genome-wide signatures of DNA methylation from methylation maps across fives tissues from a male and female koala (Phascolarctos cinereus) and present the first whole genome, multi-tissue marsupial “methylome atlas.” Using these novel data, we elucidate divergent versus common features of marsupial and eutherian DNA methylation. First, tissue-specific differential DNA methylation in marsupials primarily occurs in gene bodies. Second, females show significant global reduction (hypomethylation) of X chromosome DNA methylation compared to males. We show that this pattern is also observed in eutherians. Third, on average, promoter DNA methylation shows little difference between male and female koala X chromosomes, a pattern distinct from that of eutherians. Fourth, the sex-specific DNA methylation landscape upstream of Rsx, the primary lncRNA associated with marsupial XCI, is consistent with the epigenetic regulation of female-(and presumably inactive X chromosome-) specific expression. Finally, we utilize the prominent female X chromosome hypomethylation and classify 98 previously unplaced scaffolds as X-linked, contributing an additional 14.6 Mb (21.5 %) to genomic data annotated as the koala X chromosome. Our work demonstrates evolutionarily divergent pathways leading to functionally conserved patterns of XCI in two deep branches of mammals.

scholarly journals Koala methylomes reveal divergent and conserved DNA methylation signatures of X chromosome regulation

2021 ◽  
Vol 288 (1945) ◽  
pp. 20202244
Author(s):  
Devika Singh ◽  
Dan Sun ◽  
Andrew G. King ◽  
David E. Alquezar-Planas ◽  
Rebecca N. Johnson ◽  
...  
Keyword(s):  
Dna Methylation ◽  
X Chromosome ◽  
Epigenetic Regulation ◽  
Specific Expression ◽  
X Chromosomes ◽  
Phascolarctos Cinereus ◽  
Male And Female ◽  
Inactive X Chromosome ◽  
Promoter Dna ◽  
Female Specific

X chromosome inactivation (XCI) mediated by differential DNA methylation between sexes is an iconic example of epigenetic regulation. Although XCI is shared between eutherians and marsupials, the role of DNA methylation in marsupial XCI remains contested. Here, we examine genome-wide signatures of DNA methylation across fives tissues from a male and female koala ( Phascolarctos cinereus ), and present the first whole-genome, multi-tissue marsupial ‘methylome atlas’. Using these novel data, we elucidate divergent versus common features of representative marsupial and eutherian DNA methylation. First, tissue-specific differential DNA methylation in koalas primarily occurs in gene bodies. Second, females show significant global reduction (hypomethylation) of X chromosome DNA methylation compared to males. We show that this pattern is also observed in eutherians. Third, on average, promoter DNA methylation shows little difference between male and female koala X chromosomes, a pattern distinct from that of eutherians. Fourth, the sex-specific DNA methylation landscape upstream of Rsx , the primary lnc RNA associated with marsupial XCI, is consistent with the epigenetic regulation of female-specific (and presumably inactive X chromosome-specific) expression. Finally, we use the prominent female X chromosome hypomethylation and classify 98 previously unplaced scaffolds as X-linked, contributing an additional 14.6 Mb (21.5%) to genomic data annotated as the koala X chromosome. Our work demonstrates evolutionarily divergent pathways leading to functionally conserved patterns of XCI in two deep branches of mammals.

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