Application of the FISH Technique to Visualize Sex Chromosomes in Domestic Cat Spermatozoa

Fluorescence in situ hybridization is a molecular cytogenetics technique that enables the visualization of chromosomes in cells via fluorescently labeled molecular probes specific to selected chromosomes. Despite difficulties in carrying out the FISH technique on sperm, related to the need for proper nuclear chromatin decondensation, this technique has already been used to visualize chromosomes in human, mouse, cattle, swine, horse, and dog spermatozoa. Until now, FISH has not been performed on domestic cat sperm; therefore, the aim of this study was to visualize sex chromosomes in domestic cat sperm. The results showed the presence of X and Y chromosomes in feline spermatozoa. The procedure used for sperm decondensation and fluorescence in situ hybridization was adequate to visualize chromosomes in domestic cat spermatozoa and, in the future, it may be used to determine the degree of chromosomal abnormalities in these gametes.

Combination of Direct Viable Count and Fluorescent In Situ Hybridization (DVC-FISH) as a Potential Method for Identifying Viable Vibrio parahaemolyticus in Oysters and Mussels

Vibrio parahaemolyticus is a human food-borne pathogen with the ability to enter the food chain. It is able to acquire a viable, non-cultivable state (VBNC), which is not detected by traditional methods. The combination of the direct viable count method and a fluorescent in situ hybridization technique (DVC-FISH) makes it possible to detect microorganisms that can present VBNC forms in complex samples The optimization of the in vitro DVC-FISH technique for V. parahaemolyticus was carried out. The selected antibiotic was ciprofloxacin at a concentration of 0.75 μg/mL with an incubation time in DVC broth of 5 h. The DVC-FISH technique and the traditional plate culture were applied to detect and quantify the viable cells of the affected pathogen in artificially contaminated food matrices at different temperatures. The results obtained showed that low temperatures produced an important logarithmic decrease of V. parahaemolyticus, while at 22 °C, it proliferated rapidly. The DVC-FISH technique proved to be a useful tool for the detection and quantification of V. parahaemolyticus in the two seafood matrices of oysters and mussels. This is the first study in which this technique has been developed to detect viable cells for this microorganism.

A Dual-Color Tyr-FISH Method for Visualizing Genes/Markers on Plant Chromosomes to Create Integrated Genetic and Cytogenetic Maps

In situ imaging of molecular markers on a physical chromosome is an indispensable tool for refin-ing of genetic maps and validation genome assembly at the chromosomal level. Despite tremen-dous progress in genome sequencing the plant genome assembly at chromosome level still remain a challenge. Recently developed optical and Hi-C mapping aim to assist in genome assembly. For high-confidence in the genome assembly at chromosome level more independent approaches will be required. The present study aimed to refined an ultrasensitive Tyr-FISH technique and to de-velop a reliable and easy method for in situ mapping of a short unique DNA sequences on plant chromosomes. We have carefully analyzed the critical steps of the Tyr-FISH technique to find out the reasons for the failures of using the method. It has been shown that successful visualization of marker/gene depends significantly on method of chromosome slide preparation, probe design and labelling, high stringency washing. Appropriate adjustment of these steps allowed us to detect a short DNA sequence of 1.7Kb with a frequency of 51.6%. Based on our results, we developed a reliable and simple protocol for dual-color Tyr-FISH visualization of short unique DNA sequences on plant chromosomes. New protocol allows more accurate determination of the physical distance between markers and can be applied for faster integration of genetic and cytogenetic maps.

Cytogenetic characterisation and chromosomal mapping of microsatellite and telomeric repeats in two gecko species (Reptilia, Gekkonidae) from Thailand

Studies of chromosomes of Cyrtodactylus jarujini Ulber, 1993 and C. doisuthep Kunya et al., 2014 to compare microsatellite and TTAGGG sequences by classical and molecular techniques were conducted in Thailand. Karyological typing from a conventional staining technique of C. jarujini and C. doisuthep showed diploid chromosome numbers of 40 and 34 while the Fundamental Numbers (NF) were 56 in both species. In addition, we created the chromosome formula of the chromosomes of C. jarujini showing that 2n (40) = Lsm1 + Lsm2 + Lt3 + Mm1 + Mt4 + Sm2 + Sa2 + St5 while that of C. doisuthep was 2n (34) = Lsm3 + Lm2 + Lt3 + Mm1 + Mt2 + Sm4 + Sa1 + St1. Ag-NOR staining revealed NOR-bearing chromosomes in chromosome pairs 13 and 14 in C. jarujini, and in chromosome pairs 9 and 13 in C. doisuthep. This molecular study used the FISH technique, as well as microsatellite probes including (A)20, (TA)15, (CGG)10, (CGG)10, (GAA)10, (TA)15 and TTAGGG repeats. The signals showed that the different patterns in each chromosome of the Gekkonids depended on probe types. TTAGGG repeats showed high distribution on centromere and telomere regions, while (A)20, (TA)15, (CGG)10, (CGG)10, (GAA)10 and (TA)15 bearing dispersed over the whole genomes including chromosomes and some had strong signals on only a pair of homologous chromosomes. These results suggest that the genetic linkages have been highly differentiated between the two species.

Cytogenetic characterisation and chromosomal mapping of microsatellite and telomeric repeats in two gecko species (Reptilia, Gekkonidae) from Thailand

Studies of chromosomes of Cyrtodactylus jarujini Ulber, 1993 and C. doisuthep Kunya et al., 2014 to compare microsatellite and TTAGGG sequences by classical and molecular techniques were conducted in Thailand. Karyological typing from a conventional staining technique of C. jarujini and C. doisuthep showed diploid chromosome numbers of 40 and 34 while the Fundamental Numbers (NF) were 56 in both species. In addition, we created the chromosome formula of the chromosomes of C. jarujini showing that 2n (40) = Lsm1 + Lsm2 + Lt3 + Mm1 + Mt4 + Sm2 + Sa2 + St5 while that of C. doisuthep was 2n (34) = Lsm3 + Lm2 + Lt3 + Mm1 + Mt2 + Sm4 + Sa1 + St1. Ag-NOR staining revealed NOR-bearing chromosomes in chromosome pairs 13 and 14 in C. jarujini, and in chromosome pairs 9 and 13 in C. doisuthep. This molecular study used the FISH technique, as well as microsatellite probes including (A)20, (TA)15, (CGG)10, (CGG)10, (GAA)10, (TA)15 and TTAGGG repeats. The signals showed that the different patterns in each chromosome of the Gekkonids depended on probe types. TTAGGG repeats showed high distribution on centromere and telomere regions, while (A)20, (TA)15, (CGG)10, (CGG)10, (GAA)10 and (TA)15 bearing dispersed over the whole genomes including chromosomes and some had strong signals on only a pair of homologous chromosomes. These results suggest that the genetic linkages have been highly differentiated between the two species.

Structure and significance of cytogenetic abnormalities in adult patients with Ph-negative acute lymphoblastic leukemia

Objective. To evaluate occurrence, variety, structural peculiarities and prognostic meaning of cytogenetic abnormalities in adult patients with Ph-negative acute lymphoblastic leukemia (ALL) receiving therapy according to ALL-2009 protocol. Materials and methods. The study included 115 adult patients with firstly diagnosed Ph-negative ALL: 58 male and 57 female aged from 15 to 61 years (mean age 26.5 years), who underwent treatment from September 2009 to September 2015 in National Medical Research Center for Hematology MH RF (n=101) and in hematology departments of regional hospitals (n=14). All patients received therapy of ALL-2009 protocol (ClinicalTrials.gov, NCT01193933). The median follow-up was 24.5 months (0.2-94.4 months). As a part of the study results of a standard cytogenetic assay (SCA) were analyzed and fluorescence hybridization in situ (FISH) with the use of DNA-probes was performed on archived biological material for structural changes in gene locuses MLL/t(11q23), с-MYC/t(8q24), TP53/ deletion 17p13, CDKN2A/ deletion 9p21, translocation t(1;19)/E2A-PBX1 и t(12;21)/ETV6-RUNX1; iAMP21 identification. Results. Karyotype was defined using SCA in 86% of patients. Normal karyotype was found in 48.5% of them, chromosome aberrations in 51.5% (structural changes were found in 19.2%, hyperploidy in 27.2%, and hypoploidy in 5.1%). In 17.2% of patients complex karyotype abnormalities were found. With the use of FISH technique aberrations were found in 67% of patients: 9p21/CDKN2A deletion in 24.3%, MLL/t(11q23) gene abnormalities in 7.8%, 17p13/TP53 deletion in 5.2%, abnormalities of c-MYC/t(8q24) in 1.7%, t(1;19)/E2A-PBX1 in 0.8%, and iAMP21 in 0.8%, other abnormalities (additional signals/absence of signals from gene locuses) in 26.4%, t(12;21)/ETV6-RUNX1 was not found. FISH technique use in addition to SCA allows to increase aberrant karyotype location from 51.5 to 67%. A statistically significant correlation of 9p21/CDKN2A deletion with high serum lactate dehydrogenase activity (p=0.02); MLL/t(11q23) gene abnormalities - with leucocytosis and high blast cells level in blood (p=0.0016), hyperploidy - with normal leukocyte count (p=0.02) was shown. In groups with different cytogenetic abnormalities no statistically significant differences of treatment with ALL-2009 protocol were found (in terms of complete remission, early mortality and treatment resistance). When connection of cytogenetic abnormalities and their combinations with long-term results were analyzed according to ALL-2009 protocol, only two characteristics - MLL/t(11q23) and c MYC/t(8q24) gene abnormalities had a statistically significant influence on disease-free survival (HR - 176.9; p

EFEKTIVITAS TEKNIK MANGKUK IKAN ATAU AKUARIUM (FISH BOWL) UNTUK MENINGKATKAN HASIL BELAJAR IPS PADA SISWA KELAS V A SDN 16 CAKRANEGARA TAHUN PELAJARAN 2016/2017

Background of this research is the low social learning outcomes on students fifth grade at SDN 16 Cakranegara, it is caused by learning process focus on teaccher (Teacher Centered) and they used monoton method. Teacher usually teach only usuhg speech method and they didn’t support by the appropriate facilities. The problem in this research is how the efectivness of bowl fish teachnique could improve social learning outcomes on fifth grade students at SDN 16 Cakranegara in 2016 to 2017 ? The purpose is to improve social learning outcomes on fifth grade students in 2016 to 2017 by using bowl fish technique or aquarum. This is a base calssrom research in two cydes each cycle consist of planning, implementation, action, observation,and evaluation, reflection as well. The method collecting data are observation, documentation and test. In the frist cycle the result is on average 75,375 with percentage 70,83% and the scores for activities of students are 53 categories active enough. In second cycle the students having an increase on average 82,791 categories active. The conelusion is the efectivness of bowl fish technique could improve social learing outcomes on fifth grade students at SDN 16 Cakranegara in 2016 to 2017. Key Word : The efectivness, techniques of “bowl fish technique or aquarium”, and social learing out comes. Abstrak Penelitian ini dilatarbelakangi rendahnya hasil belajar IPS siswa kelas V A di SDN 16 Cakranegara. Hal ini disebabkan pembelajaran masih terpusat pada guru (Teacher Centered) dan teknik serta metode yang digunakan monoton. Guru biasanya hanya mengajar dengan metode ceramah saja, dan tidak di dukung dengan sarana dan prasarana yang memadai. Masalah penelitian ini adalah bagaimanakah Efektivitas Teknik Mangkuk Ikan atau Akuarium (Fish Bowl) dapat meningkatkan hasil belajar IPS siswa kelas V A SDN 16 Cakranegara Tahun Pelajaran 2016/2017?. Tujuan penelitian ini adalah untuk meningkatkan hasil belajar IPS siswa kelas V A SDN 16 Cakra Negara Tahun Pelajaran 2016/2017 dengan menerapkan teknik Mangkuk Ikan Atau Akuarium (Fish Bowl). Jenis penelitian ini adalah penelitian tindakan kelas yang dilaksanakan dalam 2 siklus. Setiap siklus terdiri dari tahap perencanaan, pelaksanaan, tindakan, observasi, dan evaluasi serta refleksi. Metode yang digunakan dalam mengumpulkan data dalam penelitian ini adalah metode observasi, dokumentasi dan tes. Pada siklus I diperoleh nilai rata-rata 75,375 dengan ketuntasan klasikal 70,83% dan skor rata-rata aktivitas siswa 53 dengan kategori cukup aktif. Pada siklus II data hasil belajar dan aktivitas siswa mengalami peningkatan yakni rata-rata 82,791 dengan ketuntasan klasikal 87,5% dan skor rata-rata aktivitas belajar siswa 58,5 dengan kriteria aktif. Dengan demikian dapat disimpulkan bahwa Efektivitas Teknik Mangkuk Ikan atau Akuarium (Fish Bowl) dapat meningkatkan hasil belajar IPS siswa kelas V A SDN 16 Cakranegara Tahun Pelajaran 2016/2017. Kata Kunci: Efektivitas, Teknik “Mangkuk Ikan atau Akuarium (Fish Bowl)”, hasil belajar IPS. References Asmani, M. Jamal. 2011. 7 Tips Aplikasi Pakem. Jogjakarta: DIVA Press. Aries, E. 2011. Assesmen Dan Evaluasi. Yogyakarta: Adytia Media Publising. Arikunto, S. 2010. Prosedur Penelitian. Jakarta: Rineka Cipta. Azizah R. Anindita. 2014. Penggunaan Metode Active Learning Tipe Card Sort Untuk Meningkatkan Keaktifan dan Prestasi Belajar IPS Siswa Kelas IV SD Negeri Sendangsari. Skripsi. Pendidikan Guru Sekolah Dasar Universitas Negeri Yogyakarta. https://www.google.com, di akses pada tanggal 25 januari 2017. Alma, Buchari, dkk. 2014. Guru Professional. Bandung: Alfabeta. Huda, Miftahul. 2015.Cooperative Learning. Yogyakarta: Pustaka Pelajar. Khotimah, Husnil. 2011. Penerapan Metode Pembelajaran Fishbowl Untuk Meningkatkan Aktivitas Belajar Sains Pada Siswa Kelas IV SDN 002 Pantai Cermin Kecamatan Tapung Kabupaten Kampar. Skripsi. Pendidikan Guru Madrasah Ibtidaiyah Universitras Islam Negeri Sultan Syarif Kasim Riau. https://www.google.com, diakses tanggal 17 Desember 2016. Nurkancana, Wayan dan Sunartana. 1990. Evaluasi Hasil Belajar. Surabaya: Usaha Nasional. Qur’anni,E Afriliya. 2013. Pengaruh Metode Team Quiz Terhadap Minat Belajar dan Pencapaian Kompetensi Menghadapi Situasi Darurat Pada Mata Pelajaran K3lh Di Smk Negeri 2 Godean. Skripsi. Program Studi Pendidikan Teknik Busana Universitas Negeri Yogyakarta. https://www.google.com, diakses tanggal 25 januari 2017. Sanjaya, Wina. 2006. Strategi Pembelajaran. Jakarta: Kencana. Sari, Melda, Supriyadi, Dan Sudirman A. 2014. Penerapan Strategi Active Learning Tipe Everyone Is Ateacher Here Untuk Meningkatkan Hasil Belajar. Jurnal. Fakultas Keguruan Dan Ilmu Pendidikan Universitas Lampung. https://www.google.com, diakses tanggal 25 januari 2017. Silberman, Mel. 2010. 101 Cara Pelatihan & Pembelajaran Aktif. Jakarta Barat: Indeks. Sudjana, Nana. 2014. Dasar-Dasar Proses Belajar Mengajar. Bandung: Sinar Baru Algensindo. Suprijono, Agus. 2014. Cooperative Learning. Yogyakarta: Pustaka Pelajar. Susanto. 2013. Teori Belajar & Pembelajaran Di Sekolah Dasar. Jakarta: Kencana. Trianto. 2011. Model Pembelajaran Inovatif Berorientasi Kontroktivis. Jakarta: Prestasi Pustaka Uno, B Hamzah dan Mohamad, Nurdin. 2012. Belajar dengan Pendekatan PAILKEM.Jakarta: Bumi Aksara. Utami, B. Rahmawati. 2012. Keefektifan Metode Fishbowl Terhadap Pembelajaran Berdiskusi Pada Siswa Kelas Viii Smp Negeri 5 Sleman. Skripsi. Pendidikan Bahasa Dan Sastra Indonesia Universitas Negeri Yogyakarta https://www.google.com, diakses tanggal 26 november 2016. Wahidah, S, Susan. 2014. Penerpan Metode “STEAVES BERHTO” (STUDENT TEAM-ACHIEVEMENT DIVISIONS DAN NUMBERED HEADS TOGETHER) untuk meningkatkan hasil belajar IPS siswa keas IVC SDN 1 kediri Tahun Pelajaran 2013/2014. Skripsi. Keguruan dan Ilmu Pendidikan, Pendidikan Guru Sekolah Dasar. Universitas Mataram. Warsono dan Hariyanto. 2013. Pembelajaran Aktif. Bandung: Remaja Rosdakarya.

Detection of PAX8/PPAR fusion gene by FISH Technique in follicular thyroid lesions

The main problem established by a discovery of a thyroid nodule is to discriminate between a benign and malignant lesion. Differential diagnosis between follicular thyroid cancer (FTC) and benign follicular thyroid adenoma (FTA) is a great challenge for even an experienced pathologist and requires special effort.The purpose of the study was to detect PAX8/PPAR G gene rearrangement by fluorescent in situ hybridization (FISH) technique,the present study is the first time in Iraq used this technique to detect fusion gene in follicular thyroid lesions,follicular carcinoma (FC) follicular variant papillary of carcinoma (FVPC), follicular adenoma (FA) and follicular hyperplasia. A total of 120 paraffin block were included in the study,30 blocks were (FC),30 blocks were (FVPC). 30 blocks were (FA),30 blocks were blocks thyroid follicular hyperplasia.20 blocks endocervical epithelium.20 paraffin blocks of colonic epithelium. The clinicopathological parameters were obtained from patients’ admission case sheets and pathology reports (age,gender). The description of measures regarding fluorescent instu hybridization of PAX8/ PPAR G fusion gene there was significantly highest in follicular carcinoma,followed by follicular variant of papillary carcinoma,then follicular adenoma and lastly by follicular hyperplasia The area under the curve for all three markers was >0.8.The p-value was highly significant for all three markers (P<0.001). Cutoff values that predict malignant thyroid follicles lesion were as following: score of >1 for Galectin-3 marker,score of >2 for HBME-1 marker and number of positive cells per 50 of >13 (>26%) for PAX8 /PPAR G fusion gene marker. The sensitivities for the three markers were 100.00 %,90.00 % and 66.67 %,respectively,whereas the specificities were 88.30 %,98.33 % and 100.00 % respectively.