Reducing the metabolic burden of rRNA synthesis promotes healthy longevity in Caenorhabditis elegans.

Ribosome biogenesis is initiated by RNA polymerase I (Pol I)-mediated synthesis of pre-ribosomal RNA (pre-rRNA). Pol I activity was previously linked to longevity, but the underlying mechanisms were not studied beyond effects on nucleolar structure and protein translation. Here we use multi-omics and functional tests to show that curtailment of Pol I activity remodels the lipidome and preserves mitochondrial function to promote longevity in Caenorhabditis elegans. Reduced pre-rRNA synthesis improves energy homeostasis and metabolic plasticity also in human primary cells. Conversely, the enhancement of pre-rRNA synthesis boosts growth and neuromuscular performance of young nematodes at the cost of accelerated metabolic decline, mitochondrial stress and premature aging. Moreover, restriction of Pol I activity extends lifespan more potently than direct repression of protein synthesis, and confers geroprotection even when initiated late in life, showcasing this intervention as an effective longevity and metabolic health treatment not limited by aging.

Echocardiographic Features of the Ductus Arteriosus and the Foramen Ovale in a Hospital-Based Population of Neonatal Foals.

The ductus arteriosus (DA) and foramen ovale (FO), including the septum primum (SP) and septum secundum (SS), are important structures in fetal circulation and are unexplored in neonatal equids. The objective of this study is to describe echocardiographic characteristics in a hospital-based population of neonatal foals. On days 2, 5 and 10 after parturition, cardiac ultrasound was performed, and clinical data were collected in healthy and diseased Warmblood foals. Fifty healthy (n = 15) and diseased (n = 35) Warmblood foals were examined. A left-sided and right-sided holosystolic murmur was audible in 98% (n = 42) and 51% (n = 22), respectively, on day 2; in 81% (n = 25) and 19% (n = 6) on day 5; and in 44% (n = 4) and 11% (n = 1) on day 10. The median grade of the systolic murmurs was higher when the DA was open. Flow through the DA could be visualized with color flow and continuous wave (CW) Doppler from the left parasternal long-axis view of the pulmonary artery in 40/43 foals on day 2, 9/31 foals on day 5 and 2/9 foals on day 10. The DA diameter was 2 ± 1 mm on day 2, 2 ± 1 mm on day 5 and 1 mm on day 10. The thickness of both septa of the FO was similar. The SP fluttered into the left atrium at all ages, but the maximal distance between the SP and SS decreased over time. In conclusion, cardiac murmurs, a patent DA and fluttering FO are frequent findings in neonatal foals. While these findings are probably physiological, the clinical importance needs to be further elucidated.

Successful application of closed loop stimulation pacemakers with remote monitoring in 3 miniature donkeys with syncope.

Rate-adaptive single chamber pacemakers with accelerometer, closed loop stimulation (CLS), and remote monitoring functionality (Eluna 8 SR-T, Biotronik, SE & Co, Germany) were implanted in 3 miniature donkeys with third-degree atrioventricular block and syncope. After recovery, different pacemaker programming modes were tested at rest, during stress without physical exercise and during physical exercise. Pacing rates were compared to actual atrial rates and showed that CLS functionality allowed physiological heart rate adaptation. A transmitter installed in the stable provided wireless connection of the pacemaker to the internet. Home monitoring was activated which performed daily wireless transmission of pacemaker functional measurements to an online server allowing diagnosis of pathological arrhythmias and pacemaker malfunction from a distance. Closed loop stimulation and remote monitoring functionality resulted in nearly physiological rate adaptation and allowed remote "from-the-stable" patient follow-up.

Prevalence and characteristics of ventricular septal defects in a non-racehorse equine population (2008-2019).

BACKGROUND: Ventricular septal defects (VSDs) are the most common congenital cardiac defect in horses. OBJECTIVES: To identify prevalence, age, breed, and sex distribution of VSD and to describe associated clinical and ultrasonographic findings. ANIMALS: Hospital-based population of 21 136 horses presented to the equine internal medicine department. METHODS: Medical records over a 12-year period were reviewed for VSD confirmed by ultrasonography. Age, breed, sex, sport discipline, murmur, clinical signs, outcome, VSD type, VSD size, shunt velocity, cardiac dimensions, concomitant cardiac anomalies, and valvular regurgitations were recorded. RESULTS: From 1894 horses that underwent echocardiography, 54 had a VSD: 42 as an isolated lesion and 12 as part of complex congenital heart disease (CHD). Median age was 5 years (range, 0-26) and 1 year (range, 0-8), respectively. Warmbloods and males were overrepresented. In the isolated VSD group, only 15% had associated clinical signs and most horses had a perimembranous VSD (pmVSD; 36/42). Horses with a pmVSD and clinical signs showed a significantly lower maximal shunt velocity (3.77 vs 5.20 m/s; P < .001), higher VSD/Aortic root (Ao) diameter (0.52 vs 0.38; P = .05), higher left atrium/Ao diameter (1.94 vs 1.22; P < .001), and higher pulmonary artery/Ao diameter (1.15 vs 0.88; P = .005) compared to horses without clinical signs. All horses with complex CHD had clinical signs and abnormal cardiac dimensions. CONCLUSION AND CLINICAL IMPORTANCE: Most isolated VSD were diagnosed only at a later age and were not associated with clinical signs. Horses with complex CHD were more likely to have or develop clinical signs at younger age.

Dichotomous Impact of Myc on rRNA Gene Activation and Silencing in B Cell Lymphomagenesis.

A major transcriptional output of cells is ribosomal RNA (rRNA), synthesized by RNA polymerase I (Pol I) from multicopy rRNA genes (rDNA). Constitutive silencing of an rDNA fraction by promoter CpG methylation contributes to the stabilization of these otherwise highly active loci. In cancers driven by the oncoprotein Myc, excessive Myc directly stimulates rDNA transcription. However, it is not clear when during carcinogenesis this mechanism emerges, and how Myc-driven rDNA activation affects epigenetic silencing. Here, we have used the Eµ-Myc mouse model to investigate rDNA transcription and epigenetic regulation in Myc-driven B cell lymphomagenesis. We have developed a refined cytometric strategy to isolate B cells from the tumor initiation, promotion, and progression phases, and found a substantial increase of both Myc and rRNA gene expression only in established lymphoma. Surprisingly, promoter CpG methylation and the machinery for rDNA silencing were also strongly up-regulated in the tumor progression state. The data indicate a dichotomous role of oncogenic Myc in rDNA regulation, boosting transcription as well as reinforcing repression of silent repeats, which may provide a novel angle on perturbing Myc function in cancer cells.

IMP dehydrogenase-2 drives aberrant nucleolar activity and promotes tumorigenesis in glioblastoma.

In many cancers, high proliferation rates correlate with elevation of rRNA and tRNA levels, and nucleolar hypertrophy. However, the underlying mechanisms linking increased nucleolar transcription and tumorigenesis are only minimally understood. Here we show that IMP dehydrogenase-2 (IMPDH2), the rate-limiting enzyme for de novo guanine nucleotide biosynthesis, is overexpressed in the highly lethal brain cancer glioblastoma. This leads to increased rRNA and tRNA synthesis, stabilization of the nucleolar GTP-binding protein nucleostemin, and enlarged, malformed nucleoli. Pharmacological or genetic inactivation of IMPDH2 in glioblastoma reverses these effects and inhibits cell proliferation, whereas untransformed glia cells are unaffected by similar IMPDH2 perturbations. Impairment of IMPDH2 activity triggers nucleolar stress and growth arrest of glioblastoma cells even in the absence of functional p53. Our results reveal that upregulation of IMPDH2 is a prerequisite for the occurance of aberrant nucleolar function and increased anabolic processes in glioblastoma, which constitutes a primary event in gliomagenesis.

Survival of primary, but not of cancer cells after combined Plk1-HDAC inhibition.

In the current study we examined the combination of SAHA and SBE13 in cancer and non-cancer cells. HeLa cells displayed a synergistically reduced cell proliferation, which was much weaker in hTERT-RPE1 or NIH-3T3 cells. Cell cycle distribution differed in HeLa, hTERT-RPE1 and NIH-3T3 cells. SAHA-treated HeLa cells showed slightly increasing cell numbers in G2/M phase, but after combination with SBE13 strongly elevated cell numbers in G2/M and S phase, accompanied by decreasing G0/G1 percentages. hTERT-RPE1 and NIH-3T3 cells showed strongly enriched cell numbers in G0/G1 phase. Western blot and quantitative real time analyses revealed reduced Plk1 mRNA and protein in all cells. p21 protein was strongly induced in cancer, but not in non-cancer cells, corresponding to a different localization in immunofluorescence studies. Additionally, these revealed an abundantly present pRb protein in HeLa cells after any treatment but almost completely vanished pRb staining in treated hTERT-RPE1 cells. These differences could be approved in Western blots against Parp and Caspase 3, which were activated in HeLa, but not in hTERT-RPE1 cells. Thus, we observed for the first time a differential effect of cancer versus non-cancer cells after treatment with SAHA and SBE13, which might be due to the dual role of p21.

Vanillin-derived antiproliferative compounds influence Plk1 activity.

We synthesized a series of vanillin-derived compounds and analyzed them in HeLa cells for their effects on the proliferation of cancer cells. The molecules are derivatives of the lead compound SBE13, a potent inhibitor of the inactive conformation of human polo-like kinase 1 (Plk1). Some of the new designs were able to inhibit cancer cell proliferation to a similar extent as the lead structure. Two of the compounds ((({4-[(6-chloropyridin-3-yl)methoxy]-3-methoxyphenyl}methyl)(pyridin-4-ylmethyl)amine) and (({4-[(4-chlorophenyl)methoxy]-3-methoxyphenyl}methyl)(pyridin-4-ylmethyl)amine)) were much stronger in their capacity to reduce HeLa cell proliferation and turned out to potently induce apoptosis and reduce Plk1 kinase activity in vitro.

Combinatorial inhibition of Plk1 and PKCß in cancer cells with different p53 status.

PKCß and Plk1 are fascinating targets in cancer therapy. Therefore, we combined Enzastaurin targeting PKCß and SBE13 targeting Plk1 to test synergistic effects in cells with different p53 status. We analyzed cell proliferation and apoptosis induction, and did Western blot and FACScan analyses to examine the combined PKCß and Plk1 inhibition. p53-wild-type cells are more resistant to the combinatorial treatment than p53-deficient cells, which displayed a synergistic reduction of cell proliferation after the combination. HeLa, MCF-7 and HCT116(p53wt) and HCT116(p53-/-) cells differed in their cell cycle distribution after combinatorial treatment in dependence on a functional p53-dependent G1/S checkpoint (p53-deficient cells showed an enrichment in S and G2/M, p53-wild-type cells in G0/G1 phase). hTERT-RPE1 cells did not show the synergistic effects of cancer cells. Thus, we demonstrate for the first time that Plk1 inhibition using SBE13 enhances the effects of Enzastaurin in cancer cells. HCT116(p53wt) and HCT116(p53-/-) cells confirmed the p53-dependence of different effects after Plk1 and PKCß inhibition observed in HeLa and MCF-7 cells. Obviously, p53 protects cells from the cytotoxicity of Enzastaurin in combination with SBE13. For that reason this combination can be useful to treat p53-deficient cancers, without displaying toxicity to normal cells, which all have functional p53.

Do injection drug users have more adverse events during procedural sedation and analgesia for incision and drainage of cutaneous abscesses?

OBJECTIVE: Injection drug users (IDUs) often undergo procedural sedation and analgesia (PSA) in the emergency department (ED). We compared adverse events (AEs) for IDUs to those for non-IDUs receiving PSA for incision and drainage of cutaneous abscesses. METHODS: This was a retrospective analysis of a PSA safety audit. IDU status was prospectively documented among consecutive patients undergoing PSA at two urban EDs. Structured data describing comorbidities, vital signs, sedation regimens, and adverse events were collected. Primary outcome was the proportion of patients in each group experiencing an AE, whereas the secondary outcomes included recovery times. RESULTS: Of 525 consecutive patients receiving PSA for incision and drainage of an abscess, 244 were deemed IDUs and 281 non-IDUs. IDUs received higher doses of sedatives and analgesics, and 14 experienced AEs (5.7%), whereas 10 non-IDUs had AEs (3.6%), for a risk difference of 2.1% (95% CI -1.8, 6.5). Median recovery times were 18 minutes (interquartile range [IQR] 10-36) for IDUs and 12 minutes (IQR 7-19) for non-IDUs, for a difference of 6 minutes (95% CI 2-9 minutes). Median sedation times were also longer in IDUs, for a difference of 6 minutes (95% CI 5-10 minutes). Of 20 IDU patients and 1 non-IDU patient admitted to hospital, none had experienced an AE related to PSA. CONCLUSIONS: For ED patients requiring PSA for incision and drainage, IDUs had an AE rate similar to that of non-IDUs but longer sedation and recovery times. In experienced hands, PSA may be as safe in IDUs as in patients who do not use injection drugs.

Drugs by numbers: reaction-driven de novo design of potent and selective anticancer leads.

A potent and selective inhibitor of the anticancer target Polo-like kinase 1 was found by computer-based molecular design. This type II kinase inhibitor was synthesized as suggested by the design software DOGS and exhibited significant antiproliferative effects against HeLa cells without affecting nontransformed cells. The study provides a proof-of-concept for reaction-based de novo design as a leading tool for drug discovery.