Journal articles: 'Non-radioactive in situ hybridisation'

1

Osborne, Peter, and Peter K. Dearden. "Non-radioactive in-situ hybridisation to honeybee embryos and ovaries." Apidologie 36, no. 1 (January 2005): 113–18. http://dx.doi.org/10.1051/apido:2004075.

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Naher, H., D. Petzoldt, and K. K. Sethi. "Evaluation of non-radioactive in situ hybridisation method to detect Chlamydia trachomatis in cell culture." Sexually Transmitted Infections 64, no. 3 (June 1, 1988): 162–64. http://dx.doi.org/10.1136/sti.64.3.162.

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Brown, A., JA Hoyland, and EB Mawer. "P45. Demonstration of 1.25(OH)2D3 receptor mRNA by a novel non-radioactive in situ hybridisation method." Bone 15, no. 2 (March 1994): 241. http://dx.doi.org/10.1016/8756-3282(94)90778-1.

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Liem, R., N. Brouwer, and J. Copray. "Ultrastructural localisation of intramuscular expression of BDNF mRNA by silver-gold intensified non-radioactive in situ hybridisation." Histochemistry and Cell Biology 116, no. 6 (November 27, 2001): 545–51. http://dx.doi.org/10.1007/s00418-001-0349-z.

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Coates, P. J., W. P. Mak, G. Slavin, and A. J. d'Ardenne. "Detection of single copies of Epstein-Barr virus in paraffin wax sections by non-radioactive in situ hybridisation." Journal of Clinical Pathology 44, no. 6 (June 1, 1991): 487–91. http://dx.doi.org/10.1136/jcp.44.6.487.

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Veal, Duncan, Philip Bell, Hayley Brown, Hung-Yoon Choi, and Peter Karuso. "Fluorophores from fungi." Microbiology Australia 24, no. 3 (2003): 12. http://dx.doi.org/10.1071/ma03312.

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Fluorescence has many advantages over traditional colour and radioactive labels, and is playing an increasingly important role in the most powerful analytical techniques. For example, fluorescence is at the heart of many nucleic acid based diagnostics (e.g. DNA microarray, real time-PCR, fluorescence in situ hybridisation, etc), immunofluorescence assays, defined substrate technologies and differential display proteomics and is gradually replacing or complementing other techniques based on colour or radiolabels.

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Troncone, G., S. M. Anderson, C. S. Herrington, M. L. de Angelis, H. Noell, J. A. Chimera, and J. O'D McGee. "Comparative analysis of human papillomavirus detection by dot blot hybridisation and non-isotopic in situ hybridisation." Journal of Clinical Pathology 45, no. 10 (October 1, 1992): 866–70. http://dx.doi.org/10.1136/jcp.45.10.866.

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Keil, C., B. Husen, J. Giebel, G. Rune, and R. Walther. "Glycodelin mRNA is expressed in the genital tract of male and female rats (Rattus norvegicus)." Journal of Molecular Endocrinology 23, no. 1 (August 1, 1999): 57–66. http://dx.doi.org/10.1677/jme.0.0230057.

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In the present study we demonstrate for the first time the expression of glycodelin mRNA in the female and male genital tracts of rats using non-radioactive in situ hybridisation. Glycodelin fragment 1 (+41 to +141) shares 100% homology with the human gene sequence. In the ovary, glycodelin mRNA was restricted to granulosa cells. In the uterus, glycodelin mRNA was expressed in all epithelial cells of the endometrium. In the male reproductive tract, glycodelin mRNA was distributed in all epithelial cells of the epididymis, the prostate and the seminal vesicle. However, in the testis, glycodelin mRNA was predominantly found in spermatogonia and in spermatocytes of the seminiferous epithelium. The expression in several reproductive organs of rats offers an excellent tool to study further the physiological role of glycodelin, which is so far thought to act as an immunosuppressive factor.

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Cooper, K. "Physical state of human papillomavirus using non-isotopic in situ hybridisation." Journal of Clinical Pathology 48, no. 8 (August 1, 1995): 786. http://dx.doi.org/10.1136/jcp.48.8.786.

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10

Sinowatz, F., D. Schams, S. Kolle, A. Plath, D. Lincoln, and MJ Waters. "Cellular localisation of GH receptor in the bovine mammary gland during mammogenesis, lactation and involution." Journal of Endocrinology 166, no. 3 (September 1, 2000): 503–10. http://dx.doi.org/10.1677/joe.0.1660503.

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We have used immunohistochemistry and non-radioactive in situ hybridisation to localise the GH receptor and its transcript in the bovine mammary gland during mammogenesis, lactation and involution. We found a characteristic pattern of immunoreactive GH (irGH) receptor distribution in the epithelial and stromal compartments during the different stages of mammary gland development: The ductular epithelium showed a distinct staining for irGH receptor during most stages, whereas the alveolar epithelium contained a modest amount of GH receptor during pregnancy which increased during lactation and galactopoiesis. In dry cows, the immunostaining for GH receptors in the alveolar epithelium was very weak or negative. Curiously, the amount of GH receptor mRNA appeared relatively constant during mammogenesis and lactation. The epithelial cells of the alveoli and ducts as well as the endothelial cells showed a distinct signal in our in situ hy! bridisation studies. The predominant localisation of GH receptors in the epithelium of ducts and alveoli is supportive of a role for GH in epithelial differentiation and maintenance. Furthermore, the increased intensity of immunostaining in bovine mammary tissue post partum suggests a direct role for GH receptor in mediating the effect of GH in milk production and secretion.

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Quennell, J. H., J. L. Stanton, and P. R. Hurst. "227. FSH receptor expression in small human ovarian follicles." Reproduction, Fertility and Development 17, no. 9 (2005): 88. http://dx.doi.org/10.1071/srb05abs227.

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Follicle-stimulating hormone (FSH) is pivotal in ovarian follicle development; the granulosa cells are the targets of FSH action in the ovary via FSH receptors. Granulosa cell growth and division mark initial follicle recruitment. The acquisition of FSH receptors on granulosa cells is regarded as a key event in hormone responsiveness and consequently follicle development. Due to the low abundance of FSH receptors and low expression of its mRNA it has been difficult to definitively characterise FSH receptor expression patterns. Here, localisation of FSH receptor in different follicle populations has been assessed with in situ hybridisation and real-time PCR of laser microdissected samples. We have used non-radioactive in situ hybridisation to investigate FSH receptor mRNA on a wide range of follicle stages. Biopsies from healthy fertile women (28–33 years) were frozen, embedded and cryosectioned at 10 µm. DIG-labelled RNA probes were designed to detect all splice variants. Hybridised probes were detected with NBT/BCIP in a colorimetric reaction. Secondly, follicles of different morphometric stages were isolated with a laser microscope. RNA extraction, reverse transcription and real-time PCR were used to confirm RNA presence and quantify relative expression. All follicle stages (from primordial to large antral) showed the presence of FSH receptor mRNA in their granulosa cells; sense controls were negative. Observations from real-time PCR indicate FSH receptor mRNA is present in all follicle stages observed and relative expression levels increase over early follicle development. These results challenge the existing doctrine that FSH receptor is absent in the smallest follicles. This suggests initial follicle recruitment may involve gonadotrophins. The use of sensitive molecular techniques will be crucial in elucidating this further.

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Wiese, U. H., J. L. Ruth, and P. C. Emson. "Differential expression of growth-associated protein (GAP-43) mRNA in rat primary sensory neurons after peripheral nerve lesion: a non-radioactive in situ hybridisation study." Brain Research 592, no. 1-2 (October 1992): 141–56. http://dx.doi.org/10.1016/0006-8993(92)91669-6.

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Herrington, C. S., S. M. Anderson, H. M. Bauer, B. Troncone, M. L. de Angelis, H. Noell, J. A. Chimera, S. L. Van Eyck, and J. O. McGee. "Comparative analysis of human papillomavirus detection by PCR and non-isotopic in situ hybridisation." Journal of Clinical Pathology 48, no. 5 (May 1, 1995): 415–19. http://dx.doi.org/10.1136/jcp.48.5.415.

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14

Berrieman, H. K., J. N. E. Ashman, M. E. Cowen, J. Greenman, M. J. Lind, and L. Cawkwell. "Chromosomal analysis of non-small-cell lung cancer by multicolour fluorescent in situ hybridisation." British Journal of Cancer 90, no. 4 (February 2004): 900–905. http://dx.doi.org/10.1038/sj.bjc.6601569.

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Brackett, Diane G., Azfar Neyaz, Kshitij Arora, Ricard Masia, Anthony Mattia, Lawerence Zukerberg, Joseph Misdraji, et al. "Cholangiolar pattern and albumin in situ hybridisation enable a diagnosis of intrahepatic cholangiocarcinoma." Journal of Clinical Pathology 73, no. 1 (August 17, 2019): 23–29. http://dx.doi.org/10.1136/jclinpath-2019-206055.

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AimsThe histological distinction of intrahepatic cholangiocarcinoma (ICC) from metastatic adenocarcinoma remains a challenge. The primary goal was to evaluate the diagnostic value of morphology and albumin expression in the diagnosis of ICC.MethodsWe evaluated morphological patterns in 120 ICCs and 677 non-hepatic adenocarcinomas and performed in situ hybridisation (ISH) stain for albumin in the former cohort (retrospective cohort). We also identified 119 samples from primary and metastatic lesions, the validation cohort, in which albumin ISH was performed as part of the diagnostic workup. Targeted sequencing was performed on selected cases. We also mined existing expression profiling data including cases from The Cancer Genome Atlas (TCGA) (41 760 unique samples).ResultsIn the retrospective cohort, 45% of ICCs and <1% of non-hepatic adenocarcinomas showed a cholangiolar pattern; albumin ISH was positive in 93% of ICCs with significant intratumorous heterogeneity. In the validation cohort, 29% of ICCs showed a cholangiolar pattern and 88% expressed albumin, while all metastatic non-hepatic neoplasms were negative (n=37) (sensitivity 88% and specificity 100%). Targetable genetic alterations (IDH mutations and FGFR2 fusions) were identified in 31% of ICCs (10 of 32). An analysis of the TCGA data validated the specificity of the albumin assay.ConclusionsThe cholangiolar pattern and albumin RNA ISH distinguishes ICC from metastatic adenocarcinoma with high specificity. Given the high prevalence of targetable mutations in ICC, albumin RNA ISH is an essential component in the workup of tumours of uncertain origin. A specific diagnosis of ICC could trigger molecular testing and uncover targetable genetic alterations.

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Sayyadi, Nima, Russell E. Connally, Thomas S. Lawson, Jingli Yuan, Nicolle H. Packer, and James A. Piper. "Time-Gated Luminescent In Situ Hybridization (LISH): Highly Sensitive Detection of Pathogenic Staphylococcus aureus." Molecules 24, no. 11 (May 31, 2019): 2083. http://dx.doi.org/10.3390/molecules24112083.

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We describe simple direct conjugation of a single TEGylated Europium chelate to DNA that binds to intracellular rRNA and is then detected using a homogeneous luminescent in situ hybridisation (LISH) technique. As a proof-of-principle, Staphylococcus aureus (S. aureus) was selected as a model for our study to show the ability of this probe to bind to intracellular 16S ribosomal rRNA. A highly purified Europium chelate conjugated oligonucleotide probe complementary to an rRNA sequence-specific S. aureus was prepared and found to be soluble and stable in aqueous solution. The probe was able to bind specifically to S. aureus via in situ hybridisation to differentiate S. aureus from a closely related but less pathogenic Staphylococcus species (S. epidermidis). A time-gated luminescent (TGL) microscope system was used to generate the high signal-to-noise ratio (SNR) images of the S. aureus. After excitation (365 nm, Chelate λmax = 335 nm), the long-lived (Eu3+) luminescent emission from the probe was detected without interference from natural background autofluorescence typically seen in biological samples. The luminescent images were found to have 6 times higher SNR or sensitivity compared to the fluorescent images using conventional fluorophore Alexa Fluor 488. The TEGylated Europium chelate -oligo probe stained S. aureus with mean signal intensity 3.5 times higher than the threshold level of signal from S. epidermidis (with SNR 8 times higher). A positive control probe (EUB338–BHHTEGST–Eu3+) has mean signal intensity for S. aureus and S. epidermidis equally 3.2 times higher than the threshold of signal for a negative NON-EUB338 control probe. The direct conjugation of a single Europium chelate to DNA provides simplicity and improvement over existing bovine serum albumin (BSA)/streptavidin/biotinylated DNA platforms for multi-attachment of Europium chelate per DNA and more importantly makes it feasible for hybridisation to intracellular RNA targets. This probe has great potential for highly sensitive homogeneous in situ hybridisation detection of the vast range of intracellular DNA targets.

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Varella-Garcia, M., J. Diebold, D. A. Eberhard, K. Geenen, A. Hirschmann, M. Kockx, I. Nagelmeier, et al. "EGFR fluorescence in situ hybridisation assay: guidelines for application to non-small-cell lung cancer." Journal of Clinical Pathology 62, no. 11 (October 27, 2009): 970–77. http://dx.doi.org/10.1136/jcp.2009.066548.

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Cooper, K., C. S. Herrington, J. E. Stickland, M. F. Evans, and J. O. McGee. "Episomal and integrated human papillomavirus in cervical neoplasia shown by non-isotopic in situ hybridisation." Journal of Clinical Pathology 44, no. 12 (December 1, 1991): 990–96. http://dx.doi.org/10.1136/jcp.44.12.990.

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Zhang, Zhouwei, Donald L. Weaver, Daniel Olsen, James deKay, Zhihua Peng, Takamaru Ashikaga, and Mark F. Evans. "Long non-coding RNA chromogenic in situ hybridisation signal pattern correlation with breast tumour pathology." Journal of Clinical Pathology 69, no. 1 (August 31, 2015): 76–81. http://dx.doi.org/10.1136/jclinpath-2015-203275.

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Kurihara, Nobuyasu, Kazuhiro Imai, Hiroshi Nanjo, Ryuta Nakamura, Yuki Wakamatsu, Koji Akagami, Kaori Terata, et al. "Practical application of non-contact alternating current electric field mixing for reagent-saving in situ hybridisation of HER2." Journal of Clinical Pathology 72, no. 9 (May 25, 2019): 603–8. http://dx.doi.org/10.1136/jclinpath-2019-205830.

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AimsHuman epidermal growth factor receptor 2 (HER2)-targeted agents are effective against HER2-positive breast cancers. However, their lack of survival benefit in HER2-negative patients as well as their toxic effects and high cost highlight the need for accurate assessment of HER2 status. Our aim was to evaluate the clinical utility of a reagent-saving in situ hybridisation (Saving ISH) that facilitates hybridisation and saves HER2/chromosome enumeration probe by taking advantage of the non-contact mixing effect of an alternating current (AC) electric field.MethodsWith a new device, we apply a high-voltage, low-frequency AC electric field to the tissue sections, which mixes the probe within microdroplets as the voltage is switched on and off. Specimens (n=113) from patients with breast cancers identified immunohistochemically as HER2 0/1(+), (2+) or (3+) were used. The specimens were all tested using conventional dual ISH (DISH), DISH with an automated slide stainer (ASS) and Saving ISH (1:1–1:3 dilution).ResultsThe Saving ISH with 1:2 probe dilution produced stable results with less non-specific staining while using smaller amounts of probe. The accuracy of HER2 status with Saving ISH was equal to standard. We found 96.4% agreement between DISH using ASS and Saving ISH (kappa coefficient=0.912).ConclusionsThese results suggest reagent-saving HER2 ISH could be used as a clinical tool for accurate and stable HER2 assessment, even when reagent concentrations vary.

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Barua, S., A. Macedo, D. S. Kolb, K. E. Wynne-Edwards, and C. Klein. "Milk-fat globule epidermal growth factor 8 (MFGE8) is expressed at the embryo– and fetal–maternal interface in equine pregnancy." Reproduction, Fertility and Development 30, no. 4 (2018): 585. http://dx.doi.org/10.1071/rd17094.

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Milk-fat globule epidermal growth factor (EGF) 8 protein (MFGE8), also known as lactadherin, promotes cell adhesion in an Arg-Gly-Asp (RGD)-dependent modus via integrins. In the present study, the expression of MFGE8 was examined in equine endometrium during oestrus and at Days 12 and 16 after ovulation in pregnant and non-pregnant mares and in mares during the 5th month of gestation. Results demonstrated that MFGE8 is expressed at the embryo– and fetal–maternal interface in equine pregnancy. In non-pregnant endometrium its expression was upregulated by oestrogen, a finding that was confirmed using endometrial explant culture. MFGE8 was expressed at similar levels by conceptuses collected 13 and 14 days after ovulation and by allantochorion sampled during the 5th month of gestation. Pericytes of endometrial blood vessels displayed strong MFGE8 expression upon in situ hybridisation. During the 5th month of gestation, the fetal side of the allantochorionic villi in particular displayed pronounced staining upon in situ hybridisation, confirming that MFGE8 expression is not restricted to early pregnancy but persists and is present at the fetal–maternal interface. Potential roles of MFGE8 in equine pregnancy include mediating cell–cell adhesion, promotion of angiogenesis and placental transfer of fatty acids.

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Pollanen, R., S. Vuopala, and V. P. Lehto. "Detection of human papillomavirus infection by non-isotopic in situ hybridisation in condylomatous and CIN lesions." Journal of Clinical Pathology 46, no. 10 (October 1, 1993): 936–39. http://dx.doi.org/10.1136/jcp.46.10.936.

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Fujimoto, Yoshiko, Kenichi Nomura, Shuji Fukada, Daisuke Shimizu, Kazuho Shimura, Yosuke Matsumoto, Shigeo Horiike, et al. "Immunoglobulin light chain gene translocations in non-Hodgkin’s lymphoma as assessed by fluorescence in situ hybridisation." European Journal of Haematology 80, no. 2 (November 15, 2007): 143–50. http://dx.doi.org/10.1111/j.1600-0609.2007.00993.x.

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Hartt, L. S., S. J. Carling, M. M. Joyce, G. A. Johnson, D. K. Vanderwall, and T. L. Ott. "Temporal and spatial associations of oestrogen receptor alpha and progesterone receptor in the endometrium of cyclic and early pregnant mares." Reproduction 130, no. 2 (August 2005): 241–50. http://dx.doi.org/10.1530/rep.1.00596.

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Uterine function is primarily controlled by the combined actions of oestrogen and progesterone working through their cognate nuclear receptors. The mechanism of establishment of pregnancy in the mare is of interest because it involves prolonged pre-attachment and conceptus migration phases, and both invasive and non-invasive placental cell types, and as such has been an important comparative model. This study characterised regulation of oestrogen (ER) and progesterone (PR) receptors in the endometrium of the mare during the oestrous cycle and early pregnancy. Endometrial tissues collected during the oestrous cycle and early pregnancy were analysed for steady-state levels of ER and PR mRNA and protein. Steady-state levels of ER and PR mRNA were highest on days 0, 17 and 20 in cyclic mares and lowest on days 11 and 14. A day-by-status interaction was detected, indicating that day 17 and day 20 pregnant mares exhibited low levels of ER and PR compared with the corresponding days of the oestrous cycle. In situ hybridisation analyses showed receptor mRNA localisation primarily in the luminal epithelium (LE), glandular epithelium (GE) and stroma around oestrus. During dioestrus and early pregnancy, receptors were not detected in the LE, and were lower in the stroma and deeper GE. Changes in hybridisation intensity in these cell types were consistent with changes in mRNA levels detected by slot-blot hybridisation. ER and PR proteins were detected in the nuclei of LE, GE and stromal cells. Consistent with results from in situ hybridisation, levels of ER and PR immunoreactivity were higher around oestrus, declined to low levels during dioestrus and remained low during early pregnancy. Results described here for temporal and spatial changes in steroid receptor gene expression in mares show the greatest similarities with those described for cattle and sheep.

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Arun, C., M. DeCatris, D. M. Hemingway, N. J. M. London, and K. J. O'Byrne. "Endothelin-1 is a Novel Prognostic Factor in Non-Small Cell Lung Cancer." International Journal of Biological Markers 19, no. 4 (October 2004): 262–67. http://dx.doi.org/10.1177/172460080401900402.

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Endothelin-1 (ET-1) is a potent vasoactive peptide and a hypoxia-inducible angiogenic growth factor associated with the development and growth of solid tumours. This study evaluated the expression of big endothelin-1 (big ET-1), a stable precursor of ET-1, and ET-1 in non-small cell lung cancer (NSCLC). Big ET-1 expression was evaluated in paraffin-embedded tissue sections from 10 NSCLC tumours using immunohistochemistry and in situ hybridisation. The production of big ET-1 and ET-1 was studied in six established NSCLC cell lines. The plasma concentrations of big ET-1 were measured in 30 patients with proven NSCLC prior to chemotherapy by means of a sandwich enzyme-linked immunoassay and compared to levels in 20 normal controls. Big ET-1 immunostaining was detected in the cancer cells of all tumours studied. Using in situ hybridisation, tumour cell big ET-1 mRNA expression was demonstrated in all samples. All six NSCLC cell lines expressed ET-1, with big ET-1 being detected in three. The median big ET-1 plasma level in patients with NSCLC was 5.4 pg/mL (range 0–22.7 pg/mL) and was significantly elevated compared to median big ET-1 plasma levels in controls, 2.1 pg/mL (1.2–13.4 pg/mL) (p=0.0001). Furthermore, patients with plasma big ET-1 levels above the normal range (upper tertile) had a worse outcome (p=0.01). In conclusion, big ET-1/ET-1 is expressed by resected NSCLC specimens and tumour cell lines. Plasma big ET-1 levels are elevated in NSCLC patients compared to controls with levels >7.8 pg/mL being associated with a worse outcome. The development of selective ET-1 antagonists such as Atrasentan indicates that ET-1 may be a therapeutic target in NSCLC.

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Sayed-Ahmed, A., Margit Kulcsár, P. Rudas, and T. Bartha. "Expression and localisation of leptin and leptin receptor in the mammary gland of the dry and lactating non-pregnant cow." Acta Veterinaria Hungarica 52, no. 1 (January 1, 2004): 97–111. http://dx.doi.org/10.1556/avet.52.2004.1.10.

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Leptin and leptin receptor were studied in the mammary gland of non-pregnant dry and lactating cows. Using RT-PCR it was demonstrated that leptin and its short (Ob-Ra) and long (Ob-Rb) receptor isoforms are expressed both in the dry and the lactating mammary gland tissue. Tissue distribution of leptin and its receptor mRNA transcripts were examined by insitu hybridisation, while the leptin protein was localised by immunohistochemistry. Although in situ hybridisation is semiquantitative, our morphological data suggest that the epithelial leptin mRNA expression of the lactating gland is higher than that of the dry gland. To compare the leptin mRNA levels between dry and lactating udders competitive PCR was used, which showed no difference in leptin expression for the whole mammary tissues. The lack of difference in total leptin mRNA levels is explained by the high adipose tissue content of the dry mammary gland. Leptin and its receptor transcripts are expressed mainly in the epithelial cells of lactating cows, while in dry mammary tissue the signal is found in the stromal tissues as well. The results provide additional evidence that locally produced leptin takes part in the regulation and maintenance of mammary epithelial cell activity.

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Hromadníková, Ilona, Sultan Karamanov, Bela Houbová, Dana Hridelova, Josef Kofer, and Miluse Mrstinova. "Non-Invasive Fetal Sex Determination on Fetal Erythroblasts from the Maternal Circulation Using Fluorescence in situ Hybridisation." Fetal Diagnosis and Therapy 17, no. 4 (2002): 193–99. http://dx.doi.org/10.1159/000059369.

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Troncone, G., C. S. Herrington, K. Cooper, M. L. de Angelis, and J. O. McGee. "Detection of human papillomavirus in matched cervical smears and biopsy specimens by non-isotopic in situ hybridisation." Journal of Clinical Pathology 45, no. 4 (April 1, 1992): 308–13. http://dx.doi.org/10.1136/jcp.45.4.308.

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Duncan, David Jonathan, Michel Erminio Vandenberghe, Marietta Louise Juanita Scott, and Craig Barker. "Fast fluorescence in situ hybridisation for the enhanced detection of MET in non-small cell lung cancer." PLOS ONE 14, no. 10 (October 15, 2019): e0223926. http://dx.doi.org/10.1371/journal.pone.0223926.

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Rex, Maria, Richard Church, Kevin Tointon, and P. J. Scotting. "Combination of non-isotopic in situ hybridisation with detection of enzyme activity, bromodeoxyuridine incorporation and immunohistochemical markers." Histochemistry and Cell Biology 107, no. 6 (June 24, 1997): 519–23. http://dx.doi.org/10.1007/s004180050139.

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Stoykov, I., H. C. van Beeren, A. F. M. Moorman, V. M. Christoffels, W. M. Wiersinga, and O. Bakker. "Effect of amiodarone and dronedarone administration in rats on thyroid hormone-dependent gene expression in different cardiac components." European Journal of Endocrinology 156, no. 6 (June 2007): 695–702. http://dx.doi.org/10.1530/eje-07-0017.

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Objective: In view of their different actions on thyroid hormone receptor (TR) isoforms we set out to investigate whether amiodarone (AM) and dronedarone (Dron) have different and/or component-specific effects on cardiac gene expression. Design: Rats were treated with AM or Dron and the expression of TRα 1, TRα 2, TRβ 1 and several tri-iodothyronine (T3)-regulated genes was studied in different parts of the heart, namely the right atrium (RA), left ventricular wall (LVW) and apex. Methods: Rats were treated for 14 days with 100 mg/kg body weight AM or Dron. The expression of TRα 1, TRα 2, TRβ 1 and T3-regulated genes was studied using real-time PCR and non-radioactive in situ hybridisation. Results: AM and Dron affected TR expression in the RA similarly by decreasing TRα 1 and β 1 expression by about 50%. In the LVW, AM and Dron decreased TRβ 1 and, interestingly, AM increased TRα 1. In the apex, AM also increased TRα 2. The changes seen in T3-dependent gene expression are reminiscent of foetal reprogramming. Conclusion: Taken together, our results indicate that AM and Dron have similar effects on the expression of TR isoforms in the RA, which could partly contribute to their ability to decrease heart rate. On the other hand, the more profound effect of AM appears on TR- and T3-dependent gene expression in the left ventricle suggests foetal reprogramming.

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Vuoriranta, P., M. Männistö, and H. Soranummi. "Occurrence of Sphingomonas sp. bacteria in cold climate drinking water supply system biofilms." Water Supply 3, no. 1-2 (March 1, 2003): 227–32. http://dx.doi.org/10.2166/ws.2003.0108.

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Members of the bacterial genus Sphingomonas (recently split into four genera), belonging to α-4-subclass of Proteobacteria, were isolated and characterised from water distribution network biofilms. Water temperature in the studied network, serving 200,000 people, is less than 5°C for about five months every winter. Sphingomonads, characterised using fluorescent oligonucleotide probes and fatty acid composition analysis (FAME), were a major group of bacteria among the distribution network biofilm isolates. Intact biofilms, grown on steel slides in a biofilm reactor fed with tap water, were detected in situ using fluorescence labelled oligonucleotide probes (FISH). Hybridisation with probes targeted on α-proteobacteria and sphingomonads was detected, but FISH on intact biofilms suffered from non-specific hybridisation and intensive autofluorescence, possibly due to extracellular material around the bacterial cells attached to biofilm. These preliminary results indicate that bacteria present in the distribution network biofilms in this study phylogenetically differ from those detected in more temperate regions.

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Kokalj-Vokač, Nadja, Tine Hajdinjak, Andreja Zagorac, Alenka Erjavec-Škerget, and Rajko Kavalar. "Non-invasive bladder cancer detection by fluorescent in situ hybridization on urine samples." Acta Medico-Biotechnica 3, no. 1 (November 21, 2021): 35–40. http://dx.doi.org/10.18690/actabiomed.30.

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Purpose: To evaluate the diagnostic accuracy of the UroVysion FISH test for detecting transitional cell bladder cancer on a mixed sample of urological patients. Methods: Urine samples were collected from patients before cystoscopy. In the case of tumor identification, transurethral resection and histological verification were performed. The fluorescent in situ hybridisation (FISH), using a commercial kit (UroVysion) containing hybridization probes for chromosomes 3, 7, 17, and 9p21, was used. At least 25 morphologically abnormal cells or all cells present on the slides were analyzed. Results: Of 179 samples, 1 was infected and in 35 (20%), no cells were identified. Test sensitivity was 76.2% (95% CI 52.8–91.8) and specificity 93.6% (95% CI 88.6–96.9); as compared to the data in the literature, this result was considered significant. Most tumors had numerical chromosome aberrations. The prevalence of histologically, which verified tumors, was 11.8%. Conclusion: Price and time–consuming procedures are major obstacles for use of the UroVysion FISH test. However, neither cytology nor UroVysion FISH are 100% specific. Higher sensitivity compared to cytology, which is evident not only in case of superficial, but also of invasive tumors (like T1) and its ability to provide numerical results, are advantages which may make UroVysion FISH test useful. Thus, this test has potential for future use. cells were identified. Test sensitivity was 76.2% (95% CI 52.8–91.8) and specificity

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Besse, P., and C. L. McIntyre. "Chromosome in situ hybridisation of ribosomal DNA in Erianthus sect. Ripidium species with varying chromosome numbers confirms x = 10 in Erianthus sect. Ripidium." Genome 42, no. 2 (April 1, 1999): 270–73. http://dx.doi.org/10.1139/g98-126.

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A wheat ribosomal DNA probe was used to determine the number of rDNA-carrying chromosomes in 2 Erianthus sect. Ripidium species using FISH (fluorescent in situ hybridisation) and non-fluorescent ISH. Two and four ribosomal DNA sites were revealed in E. elephantinus (2n = 20) and E. procerus (2n = 40), respectively. This result, together with previously published data showing 6 rDNA-carrying chromosomes in E. arundinaceus (2n = 60), confirms a possible basic chromosome number of x = 10 in Erianthus sect. Ripidium.Key words: Erianthus, FISH, ISH, ribosomal DNA, Saccharum, sugarcane.

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Wojas-Krawczyk, Kamila, Paweł Adam Krawczyk, Rodryg Adam Ramlau, Justyna Szumiło, Jerzy Kozielski, Ewa Kalinka-Warzocha, Maciej Bryl, et al. "The analysis of ALK gene rearrangement by fluorescence in situ hybridisation in non-small cel lung cancer patients." Współczesna Onkologia 6 (2013): 484–92. http://dx.doi.org/10.5114/wo.2013.38758.

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Morey, A. L., H. J. Porter, J. W. Keeling, and K. A. Fleming. "Non-isotopic in situ hybridisation and immunophenotyping of infected cells in the investigation of human fetal parvovirus infection." Journal of Clinical Pathology 45, no. 8 (August 1, 1992): 673–78. http://dx.doi.org/10.1136/jcp.45.8.673.

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Crolla, J. A., N. R. Dennis, and P. A. Jacobs. "A non-isotopic in situ hybridisation study of the chromosomal origin of 15 supernumerary marker chromosomes in man." Journal of Medical Genetics 29, no. 10 (October 1, 1992): 699–703. http://dx.doi.org/10.1136/jmg.29.10.699.

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von Laffert, M., P. Schirmacher, A. Warth, W. Weichert, R. Büttner, R. M. Huber, J. Wolf, F. Griesinger, M. Dietel, and Ch Grohé. "ALK-Testing in non-small cell lung cancer (NSCLC): Immunohistochemistry (IHC) and/or fluorescence in-situ Hybridisation (FISH)?" Lung Cancer 103 (January 2017): 1–5. http://dx.doi.org/10.1016/j.lungcan.2016.11.008.

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Sanno, N., A. Matsuno, J. Itoh, K. Kakimoto, A. Teramoto, and R. Y. Osamura. "Combined non-isotopic in situ hybridisation and indirect immunohistochemical analysis of hormone production in the rat pituitary gland." Molecular Pathology 49, no. 1 (February 1, 1996): M57—M60. http://dx.doi.org/10.1136/mp.49.1.m57.

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Wagner, F., A. Streubel, A. Roth, S. Stephan-Falkenau, and T. Mairinger. "Chromogenic in situ hybridisation (CISH) is a powerful method to detect ALK-positive non-small cell lung carcinomas." Journal of Clinical Pathology 67, no. 5 (November 29, 2013): 403–7. http://dx.doi.org/10.1136/jclinpath-2013-201974.

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Marin, Irina, Efrat Ofek, Jair Bar, Nadia Prisant, Marina Perelman, Camila Avivi, Gitit Lavy-Shahaf, Amir Onn, Ruth Katz, and Iris Barshack. "MiR-21, EGFR and PTEN in non-small cell lung cancer: an in situ hybridisation and immunohistochemistry study." Journal of Clinical Pathology 73, no. 10 (February 14, 2020): 636–41. http://dx.doi.org/10.1136/jclinpath-2019-206420.

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AimsTo analyse microRNA (miR)-21 distribution and expression at the cellular level in non-small cell lung cancer (NSCLC). MiR-21 is an oncogenic microRNA overexpressed in NSCLC. In previous studies, overexpression of miR-21 was evaluated from the tumour bulk by quantitative reverse transcription PCR with results expressed on average across the entire cell population.MethodsWe used in situ hybridisation and immunohistochemistry to assess the correlation between miR-21 levels and the expression of markers that may be possible targets (epidermal growth factor reaction) or may be involved in its upregulation (phosphatase and tensin homolog (PTEN), p53). The Pearson’s χ2 tests was used to assess correlation with clinicopathological data and with miR-21 expression both in tumour and tumour stroma.ResultsCytoplasmic staining and expression of Mir-21 were detected in the tumours and in associated stromal cells. Expression was highest in the stroma immediately surrounding the tumour cells and decreased as the distance from the tumour increased. No expression of miR-21 was found in normal lung parenchyma and a significant association was found between tumour localised miR-21 and PTEN.ConclusionsPresence of miR-21 in both cell tumour and stromal compartments of NSCLC and the relationship with PTEN confirms miR-21 as a microenvironment signalling molecule, possibly inducing epithelial mesenchymal transition and invasion by targeting PTEN in the stromal compartment possibly through exosomal transport. In situ immunohistochemical studies such as ours may help shed light on the complex interactions between miRNAs and its role in NSCLC biology.

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Harper, S. J., E. Bailey, C. M. McKeen, A. S. Stewart, J. H. Pringle, J. Feehally, and T. Brown. "A comparative study of digoxigenin, 2,4-dinitrophenyl, and alkaline phosphatase as deoxyoligonucleotide labels in non-radioisotopic in situ hybridisation." Journal of Clinical Pathology 50, no. 8 (August 1, 1997): 686–90. http://dx.doi.org/10.1136/jcp.50.8.686.

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Adinolfi, M., S. Stone, and D. Moralli. "Genes and genomes: Carrier detection of deletions in female relatives of X-linked disorders by non-isotopicin situ hybridisation." BioEssays 14, no. 6 (June 1992): 421–26. http://dx.doi.org/10.1002/bies.950140614.

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Hotta, Katsuyuki, Hiroyuki Yanai, Kadoaki Ohashi, Kiichiro Ninomiya, Hiromi Nakashima, Hiroe Kayatani, Minoru Takata, and Katsuyuki Kiura. "Pilot evaluation of a HER2 testing in non-small-cell lung cancer." Journal of Clinical Pathology 73, no. 6 (December 3, 2019): 353–57. http://dx.doi.org/10.1136/jclinpath-2019-206204.

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AimsHER2-positivity pattern in the specimens of immunohistochemistry (IHC) and ﬂuorescence in situ hybridisation (FISH) has been hardly reported in non-small-cell lung cancer (NSCLC).MethodsWe evaluated the characteristics of HER2-positivity pattern in formalin-ﬁxed parafﬁn-embedded samples using IHC and FISH in 15 patients enrolled in a larger prospective cohort study to survey a HER2-positive NSCLC.ResultsAs for the immunostaining pattern, most specimens (79%) demonstrated incomplete or mixed-typed membranous immunoreactivity with heterogeneity, resembling that observed in gastric cancer rather than breast cancer. Concordance between IHC-positivity and FISH-positivity was 87.5% according to the criteria for breast cancer scoring system. On application of the gastric cancer scoring system to the examined tumours, the IHC score increased in the seven (43.8%) specimens, and the concordance between IHC positivity and FISH positivity rose to 93.8%.ConclusionsIn our pilot series, the pattern of IHC reactivity closely resembled that observed in gastric cancer rather than breast cancer.Trial registration number000017003.

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Bailey, E., M. J. Bottomley, S. Westwell, J. H. Pringle, P. N. Furness, J. Feehally, P. E. Brenchley, and S. J. Harper. "Vascular endothelial growth factor mRNA expression in minimal change, membranous, and diabetic nephropathy demonstrated by non-isotopic in situ hybridisation." Journal of Clinical Pathology 52, no. 10 (October 1, 1999): 735–38. http://dx.doi.org/10.1136/jcp.52.10.735.

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Crolla, J. A., M. Smith, and Z. Docherty. "Identification and characterisation of a small marker chromosome using non-isotopic in situ hybridisation with X and Y specific probes." Journal of Medical Genetics 26, no. 3 (March 1, 1989): 192–94. http://dx.doi.org/10.1136/jmg.26.3.192.

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Martinez-Montero, J. C., C. S. Herrington, J. Stickland, H. Sawyer, M. Evans, D. M. Flannery, and J. O. McGee. "Model system for optimising mRNA non-isotopic in situ hybridisation: riboprobe detection of lysozyme mRNA in archival gut biopsy specimens." Journal of Clinical Pathology 44, no. 10 (October 1, 1991): 835–39. http://dx.doi.org/10.1136/jcp.44.10.835.

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Morrison, N., D. A. Nickson, M. W. McBride, U. W. Mueller, E. Boyd, and R. G. Sutcliffe. "Regional chromosomal assignment of human 3-beta-hydroxy-5-ene steroid dehydrogenase to 1p13.1 by non-isotopic in situ hybridisation." Human Genetics 87, no. 2 (June 1991): 223–25. http://dx.doi.org/10.1007/bf00204189.

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Murray, C., C. McAlister, and K. Elliott. "Use of fluorescence in situ hybridisation and laser microdissection to isolate male non-sperm cells in cases of sexual assault." International Congress Series 1288 (April 2006): 622–24. http://dx.doi.org/10.1016/j.ics.2005.09.109.

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Kałużewski, Tadeusz, Grzegorz K. Przybylski, Michał Bednarek, Sławomir Glazar, Magdalena Grabiec, Adam Jędrzejczyk, Łukasz Kępczyński, et al. "The Usefulness of Cell-Based and Liquid-Based Urine Tests in Clarifying the Diagnosis and Monitoring the Course of Urothelial Carcinoma. Identification of Novel, Potentially Actionable, RB1 and ERBB2 Somatic Mutations." Journal of Personalized Medicine 11, no. 5 (April 30, 2021): 362. http://dx.doi.org/10.3390/jpm11050362.

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Bladder cancer is one of the most common cancers in global statistics. One of the issues associated with this disease is the high incidence of cases with delayed diagnosis and what factors correlate with worse treatment outcomes. A possible reason for this may be the rather limited availability of non-invasive diagnostic tools. This short communication presents a case of a 68 year old male patient after an ineffective therapy, carried on for several years with symptoms commonly associated with prostate overgrowth that masked a carcinoma in situ of the urinary bladder. Implementation of several diagnostic techniques, including urine sediment cytology, immunocytochemistry, the fluorescence in situ hybridisation technique, the Bladder EpiCheck test and whole-genome sequencing, enabled the establishment of a correct diagnosis, implementation of appropriate treatment and provision of patient-friendly monitoring. The described case emphasises the usefulness of cell-based and liquid-based urine tests in bladder cancer diagnostic procedures.

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Journal articles on the topic 'Non-radioactive in situ hybridisation'

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