Somatic chromosome number and ploidy level in three Curcuma spp. from Bangladesh

Three Curcuma L. species were investigated cytogenetically which represent diversed staining pattern of heterochromatins in interphase nuclei and prophase chromosomes with orcein staining. Curcuma longa and C. caesia were found to possess 2n = 3x = 63 somatic chromosomes whereas 2n = 2x = 42 chromosome number in C. zedoaria is reported for the first time from Bangladesh. Total chromosome length recorded in C. longa, C. caesia and C. zedoaria were 145.08 ± 2.85 μm, 164.93 ± 4.29 μm and 97.78 ± 2.41 μm, respectively. This was the first attempt to measure the length of the chromosomes for these species. The experiment confirmed the basic chromosome number x = 21 with triploid (C. longa, C. caesia) and diploid (C. zedoaria) Curcuma plants. Polyploidy could be employed in the evolution and diversification of the genus Curcuma, which is an essential factor to characterize the species of this genus. Dhaka Univ. J. Biol. Sci. 30(2): 133-140, 2021 (July)

COMPARATIVE STUDY OF EFFICACY OF PAPANICOLAOU AND ACETO-ORCEIN STAINS IN DEMONSTRATING BARR BODIES IN BUCCAL MUCOSAL SMEARS

Introduction: Establishing one’s sexual identity has a critical role in medical conditions like ambiguous genitalia and in crime investigations. There are various investigations to determine the sexual identity of a person and Barr body evaluation in buccal smears forms the first line of investigation. Barr body estimation in buccal mucosal scrapes has been demonstrated by using different stains like Papanicolaou, Aceto-Orcein(AO), Feulgen, Guard, Cresyl violet, Carbol fuchsin and fluorescent staining methods, but with varying efficacy. The current study is done to evaluate the efficacy of Papanicolaou and AO stains in demonstration of Barr bodies. Materials and Methods: A total of 207 medical students were included in the study. Two buccal smears were collected from each student and subsequently, one was stained with Aceto-Orcein by squash technique and the other with Papanicolaou stain. Both slides were evaluated for percentage of Barr bodies using 1000x magnification and cytomorphological features in 400x. Results: The percentage of Barr bodies in AO stained slides ranged from 5-18 among females and 0-8 in males, while with Papanicolaou stain the ranges recorded were 4 – 12 in females, 0 – 2 in males. The accuracy of AO and Papanicolaou stains for detecting sex accurately were 97% and 91% respectively. Evaluation of the buccal smears was better in AO stained smears because of the clean background and better cytoplasmic and nuclear contrast in comparison to PAP stain. Conclusion: Aceto-Orcein staining method is rapid, economical, accurate, reproducible and comparable to Papanicolaou staining, for the detection of Barr body in buccal mucosal smears. Keywords: Aceto-Orcein, Papanicolaou, Buccal smears, Barr body.

INDEKS MITOSIS PUCUK DAUN Hibiscus rosa-sinensis L. VARIASI SINGLE PINK PADA BEBERAPA VARIASI WAKTU

AbstrakHibiscus rosa-sinensis L. atau kembang sepatu merupakan tanaman hias yang memiliki banyak manfaat dan merepresentasikan sifat poliploidi. Tujuan penelitian ini yaitu untuk mengetahui Indeks Mitosis (IM) dan jumlah kromosom pucuk daun H. rosa-sinensis pada beberapa variasi waktu. Indeks Mitosis dan waktu pengambilan pucuk sangat diperlukan untuk studi kromosom karena pada tahap tersebut karakter-karakter kromosom dapat diamati dengan jelas dan mudah dihitung. Waktu pengambilan pucuk yang dilakukan yaitu pada 08:00, 10:00, 12:00, 14:00, 16:00 WIB. Pembuatan sediaan kromosom dilakukan menggunakan metode squash menggunakan pewarna Aceto-orcein. Tahapan perlakuan meliputi perendaman pucuk daun di dalam air dingin selama 3 jam, fiksasi dalam larutan Carnoy selama ±24 jam, dan hidrolisis dalam larutan HCl 5N selama 30 menit. Hasil penelitian menunjukkan nilai IM tertinggi meristem pucuk daun H. rosa-sinensis variasi single pink besar muncul pada pukul 10:00 sebesar 94%. Berdasarkan hasil tersebut dapat disimpulkan bahwa waktu sampling yang optimal untuk analisis kromosom H. rosa-sinensis L. variasi single pink besar adalah pukul 10:00 dengan jumlah kromosom 2n= ca. 69-111. Hasil penelitian ini dapat dijadikan dasar untuk studi kromosom selanjutnya serta acuan untuk sampling variasi lainnya.Abstract Hibiscus rosa-sinensis L. is an ornamental plant that has many benefits and represents the character of polyploidy. The purpose of this study is to find out the Mitotic Index of leaf shoots Hibiscus rosa-sinensis on several shoots sampling times. The Mitotic Index and the timing of shoots sampling time are very necessary for chromosome studies because at this stage chromosomes characters can be clearly observed and easily calculated. Period time of collection the leaf shoots is from 08:00 AM to 16:00 PM, with two hours interval each at 08:00, 10:00, 12:00, 14:00, 16:00 . The chromosome preparation was carried out by using the squash method and aceto-orcein staining. The treatment steps included soaking the leaf shoots in cold water for 3 hours, fixation in Carnoy solution for ± 24 hours, and hydrolysis in 5N HCl solution for 30 minutes. The results showed that chromosomes were clearly visualized during the phase with the highest Mitotic Index. In addition, the percentage of Mitotic Index was found to be in line with the percentage of cells in late prophase. Among several sampling time variations, the highest Mitotic Index of Hibiscus rosa-sinensis leaf shoots appeared at 10:00 at 94% with the chromosome numbers of 2n= ca. 69-111. According to the data obtained, it is concluded that 10 AM is the most optimum sampling time that can be used as the basic information for further chromosome studies.

Karyomorphological study of Enhydra fluctuans Lour. (Asteraceae)

In the present study, karyomorphological features of Enhydra fluctuans Lour. were investigated. Orcein-staining was used for the authentic chromosomal characterization and identification. In E. fluctuans ‘Simple Chromocenter Type’ of interphase nuclei was observed with many small heterochromatin blocks after orcein staining. Prophase chromosomes showed ‘Gradient Type’ with orcein staining. This species was found to possess 2n=10x=110 with basic chromosome number x=11 and this is probably the first report for this species. The centromeric formula of this species was ‘38m+70sm+2ac’. The above features indicated that E. fluctuans is an advance polyploid with asymmetric karyotype. The total chromatin length (2n) was 295.92 μm. Individual chromosome length ranged from 1.84-4.26 μm. The relative length of each chromosome ranged from 0.006-0.014. The above mentioned data will be useful for karyomorphological characterization of E. fluctuans from Bangladesh. Bangladesh J. Sci. Ind. Res.54(4), 283-288, 2019

Cytotaxonomy of four Floral Variants of Impatiens Balsamina L.

Karyomorphological characters were investigated in four floral variants of Impatiens balsamina L. viz. white, pink, violet and red for proper characterization and inferring relationships among them. All four floral variants of I. balsamina were found to possess 2n = 14 chromosomes, and ‘Simple Chromocenter Type’ of interphase nuclei was observed in all variants with many small heterochromatin blocks after orcein staining. ‘Continuous Type’ of prophase chromosomes was found in red variant, whereas both ‘Continuous and Gradient Type’ were found in other three variants. The red variant could easily be distinguished from the remaining three variants by total chromosome length, centromeric formula, average chromosome length, total form value, and karyotype asymmetry plus symmetry index. Based on karyomorphology, the white, pink and violet variants of I. balsamina were found very closely related and constituted with similar genome, whereas red variant was found to possess slightly modified genome and distinct from other three floral variants. Journal of Bangladesh Academy of Sciences, Vol. 43, No. 1, 1-9, 2019

Cytogenetical characterization of Acalypha indica L. in Bangladesh

Acalypha indica L. was cytogenetically characterized after staining with orcein, CMA and DAPI. In this species "Simple Chromocenter Type" of interphase nuclei was observed with a few small heterochromatin blocks following orcein staining. Prophase chromosomes showed “Interstitial Type” of staining pattern with orcein which indicated the tendency of aggregation of heterochromatin in interstitial regions of chromosome. This plant was found to possess 2n = 20 metacentric chromosomes. The total length of 2n chromosome complement was 55.33 μm. Individual chromosome length ranged from 1.84 to 3.50 μm. The relative length of each chromosome ranged from 0.03 to 0.06. After staining with CMA, three bright bands were observed at different locations of chromosomes. A total of four DAPI-positive bands were also found of which two bands were centromeric and other two were entirely fluoresced with DAPI. The results of the cytological investigation may be useful for future characterization of this plant species. Dhaka Univ. J. Biol. Sci. 27(2): 183-189, 2018 (July)

Karyomorphology of Justicia adhatoda L. by differential staining

Karyomorphological features of Justicia adhatoda L. were investigated in this study after differential staining with orcein, CMA and DAPI for proper characterization. In this species "Simple Chromocenter Type" of interphase nuclei was observed with many small heterochromatin blocks after orcein staining. Prophase chromosomes showed "Interstitial Type" of staining pattern which indicated the tendency of aggregation of heterochromatin in interstitial regions of chromosome. This species was found to possess 2n = 34 metacentric chromosomes. The total length of 2n chromosome complement was 132.17 μm. Individual chromosome length ranged from 2.57 to 5.43 μm. The relative length of each chromosome ranged from 0.02 to 0.04. A total of seven CMA-positive bands were observed in this species whereas no band was found after DAPIstaining. This result suggested the absence of AT-rich portions and occurrence of GC-rich portions in the respective banded chromosomes. Except 2n chromosome number, the other karyological and fluorescent banding information probably was the first report for this species and it might help to enhance the chromosomal data base with detailed karyomorphological information of this plant species in Bangladesh. Dhaka Univ. J. Biol. Sci. 27(2): 175-181, 2018 (July)

Comparison of Histochemical Stainings in Evaluation of Liver Fibrosis and Correlation with Transient Elastography in Chronic Hepatitis

Background and Aim. The best staining to evaluate liver fibrosis in liver hepatitis is still a debated topic. This study aimed to compare Masson’s trichrome (MT), Sirius Red (SR), and orcein stainings in evaluating liver fibrosis in chronic HCV hepatitis (CHC) with semiquantitative and quantitative methods (Collagen Proportionate Area (CPA) by Digital Image Analysis (DIA)) and correlate them with transient elastography (TE).Methods. Liver stiffness evaluation of 111 consecutive patients with CHC was performed by TE. Semiquantitative staging by Metavir score system and CPA by DIA were assessed on liver biopsy stained with MT, SR, and orcein.Results. MT, SR, and orcein staining showed concordant results in 89.6% of cases in staging CHC, without significant difference in both semiquantitative and quantitative evaluations of fibrosis. TE values were concordant with orcein levels in 86.5% of the cases and with MT/RS in 77.5% (P<0.001). No significant correlation between the grade of necroinflammatory activity and TE values was found.Conclusion. In CHC, SR/MT and orcein stainings are almost concordant and when discordant, orcein staining is better related to TE values than MT/RS. This suggests that elastic fibers play a more important role than reticular or collagenous ones in determining stiffness values in CHC.