18F-fluorodeoxyglucose positron emission tomography findings of peripheral nerve sheath tumour of the nasal cavity in a dog.

An 8-year-old Miniature Poodle presented with chronic sneezing and unilateral epistaxis. A left-sided intranasal mass was identified on computed tomography. 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) was performed to evaluate the metabolic activity of the mass. The intranasal mass showed mildly increased 18F-FDG uptake. The maximal and mean standardized uptake values (SUVs) of the mass were 3.4 and 2.6, respectively. The maximal SUV of the mass/mean SUV of the normal liver was 2.5. The 7-cm soft, pink mass was easily removed through rhinoscopy, with subsequent dramatic improvement in clinical signs. Histopathological and immunohistochemical analyses determined that the mass was an intermediate-grade malignant peripheral nerve sheath tumour (PNST). This is the first report of 18F-FDG PET findings in a PNST in dogs.

Cross-correlated relaxation rates provide facile exchange signature in selectively labeled RNA.

Gerhard Wagner has made numerous contributions to NMR spectroscopy, particularly his developments in the field of spin-relaxation stand out in directly mapping the spectral density functions of proteins. He and his group developed experimental techniques to reveal the importance of dynamics to protein biological function and drug discovery. On his 75th birthday, we take this opportunity to highlight how some of those seminal ideas developed for proteins are being extended to RNAs. The role of dynamics in the structure and function of RNA has been a major interest in drug design and therapeutics. Here we present the use of cross-correlated relaxation rates (ηxy) from anti-TROSY (R2α) and TROSY (R2ß) to rapidly obtain qualitative information about the chemical exchange taking place within the bacterial and human A-site RNA system while reducing the sets of relaxation experiments required to map dynamics. We show that ηxy correlates with the order parameter which gives information on how flexible or rigid a residue is. We further show R2ß/ηxy can rapidly be used to probe chemical exchange as seen from its agreement with Rex. In addition, we report the ability of R2ß/ηxy to determine chemical exchange taking place within the bacterial A-site RNA during structural transitions at pH 6.2 and 6.5. Finally, comparison of the R2ß/ηxy ratios indicates bacterial A-site has greater R2ß/ηxy values for G19 (1.34 s-1), A20 (1.38 s-1), U23 (1.63 s-1) and C24 (1.51 s-1) than human A-site [A19 (0.76 s-1), A20 (1.01 s-1), U23 (0.74 s-1) and C24 (0.71 s-1)]. Taken together, we have shown that the chemical exchange can quickly be analyzed for RNA systems from cross-correlated relaxation rates.

Case Report: Technetium-99m Pertechnetate Scintigraphy Findings in a Dog With Iodine Deficiency-Induced Goitrous Hypothyroidism.

There is only one previous report of canine goitrous hypothyroidism caused by iodine deficiency from 1986. The present case report describes the novel diagnostic methods and long-term outcomes of a dog diagnosed with goitrous hypothyroidism caused by iodine deficiency. A 4-year-old neutered, female Pomeranian dog presented with a cervical mass, lethargy, and inactivity. The dog had a history of eating home-cooked diets sold by a private seller for 1 year. The physical examination and ultrasonography showed two bilaterally symmetric masses in the mid-cervical area (left, 1.8 × 1.4 cm; right, 2.3 × 1.8 cm), and they were suspected to be the thyroid glands. To identify the function of the thyroid gland, the basal concentrations of thyroid hormones [total T4 (tT4) and thyroid-stimulating hormone (TSH)] were measured and a TSH stimulation test was performed: baseline tT4, 0.5 µg/dL (reference interval, 1-4 µg/dL), baseline TSH, 0.81 µg/dL (reference interval, 0.05-0.42 µg/dL), and post-tT4, 1 µg/dL (6 h after the injection of TSH). The values indicated primary hypothyroidism. The urinary iodine concentration was 302 µg/L, which was markedly lower than that of normal dogs (1,289 µg/L). Thyroid scintigraphy with technetium-99m pertechnetate was also performed to quantify the activity of the thyroid gland, and the thyroid-to-salivary ratio was 3.35. Based on the results of these examinations and patient history, the dog was diagnosed with diet-induced (iodine deficiency) goitrous hypothyroidism. The dog was treated with iodine (62.5 µg/day). At 31 days after treatment, clinical signs and thyroid hormones were normalized (tT4, 1.3 µg/dL; TSH, 0.24 µg/dL). One year after treatment, the dog was well with normal concentrations of thyroid hormones (tT4, 1.8 µg/dL; TSH, 0.27 µg/dL) and a partially reduced goiter (left, 1.6 × 1.1 cm; right, 1.2 × 0.9 cm). This is the first case to describe novel diagnostic methods and long-term outcomes of a dog diagnosed with goitrous hypothyroidism caused by iodine deficiency.

Synthesis of [7-15N]-GTPs for RNA structure and dynamics by NMR spectroscopy.

Several isotope-labeling strategies have been developed for the study of RNA by nuclear magnetic resonance (NMR) spectroscopy. Here, we report a combined chemical and enzymatic synthesis of [7-15N]-guanosine-5'-triphosphates for incorporation into RNA via T7 RNA polymerase-based in vitro transcription. We showcase the utility of these labels to probe both structure and dynamics in two biologically important RNAs. Supplementary Information: The online version contains supplementary material available at 10.1007/s00706-022-02892-1.

Deleterious effects of carbon-carbon dipolar coupling on RNA NMR dynamics.

Many regulatory RNAs undergo dynamic exchanges that are crucial for their biological functions and NMR spectroscopy is a versatile tool for monitoring dynamic motions of biomolecules. Meaningful information on biomolecular dynamics requires an accurate measurement of relaxation parameters such as longitudinal (R1) rates, transverse (R2) rates and heteronuclear Overhauser effect (hNOE). However, earlier studies have shown that the large 13C-13C interactions complicate analysis of the carbon relaxation parameters. To investigate the effect of 13C-13C interactions on RNA dynamic studies, we performed relaxation measurements on various RNA samples with different labeling patterns and compared these measurements with the computational simulations. For uniformly labeled samples, contributions of the neighboring carbon to R1 measurements were observed. These contributions increased with increasing magnetic field and overall correlation time ([Formula: see text]) for R1 rates, necessitating more careful analysis for uniformly labeled large RNAs. In addition, the hNOE measurements were also affected by the adjacent carbon nuclei. Unlike R1 rates, R1ρ rates showed relatively good agreement between uniformly- and site-selectively labeled samples, suggesting no dramatic effect from their attached carbon, in agreement with previous observations. Overall, having more accurate rate measurements avoids complex analysis and will be a key for interpreting 13C relaxation rates for molecular motion that can provide valuable insights into cellular molecular recognition events.

Selective Small-Molecule Targeting of a Triple Helix Encoded by the Long Noncoding RNA, MALAT1.

Metastasis-associated lung adenocarcinoma transcript 1 ( Malat1/ MALAT1, mouse/human), a highly conserved long noncoding (lnc) RNA, has been linked with several physiological processes, including the alternative splicing, nuclear organization, and epigenetic modulation of gene expression. MALAT1 has also been implicated in metastasis and tumor proliferation in multiple cancer types. The 3' terminal stability element for nuclear expression (ENE) assumes a triple-helical configuration that promotes its nuclear accumulation and persistent function. Utilizing a novel small molecule microarray strategy, we identified multiple Malat1 ENE triplex-binding chemotypes, among which compounds 5 and 16 reduced Malat1 RNA levels and branching morphogenesis in a mammary tumor organoid model. Computational modeling and Förster resonance energy transfer experiments demonstrate distinct binding modes for each chemotype, conferring opposing structural changes to the triplex. Compound 5 modulates Malat1 downstream genes without affecting Neat1, a nuclear lncRNA encoded in the same chromosomal region as Malat1 with a structurally similar ENE triplex. Supporting this observation, the specificity of compound 5 for Malat1 over Neat1 and a virus-coded ENE was demonstrated by nuclear magnetic resonance spectroscopy. Small molecules specifically targeting the MALAT1 ENE triplex lay the foundation for new classes of anticancer therapeutics and molecular probes for the treatment and investigation of MALAT1-driven cancers.