A non-coding RNA balancing act: miR-346-induced DNA damage is limited by the long non-coding RNA NORAD in prostate cancer.

BACKGROUND: miR-346 was identified as an activator of Androgen Receptor (AR) signalling that associates with DNA damage response (DDR)-linked transcripts in prostate cancer (PC). We sought to delineate the impact of miR-346 on DNA damage, and its potential as a therapeutic agent. METHODS: RNA-IP, RNA-seq, RNA-ISH, DNA fibre assays, in vivo xenograft studies and bioinformatics approaches were used alongside a novel method for amplification-free, single nucleotide-resolution genome-wide mapping of DNA breaks (INDUCE-seq). RESULTS: miR-346 induces rapid and extensive DNA damage in PC cells - the first report of microRNA-induced DNA damage. Mechanistically, this is achieved through transcriptional hyperactivation, R-loop formation and replication stress, leading to checkpoint activation and cell cycle arrest. miR-346 also interacts with genome-protective lncRNA NORAD to disrupt its interaction with PUM2, leading to PUM2 stabilisation and its increased turnover of DNA damage response (DDR) transcripts. Confirming clinical relevance, NORAD expression and activity strongly correlate with poor PC clinical outcomes and increased DDR in biopsy RNA-seq studies. In contrast, miR-346 is associated with improved PC survival. INDUCE-seq reveals that miR-346-induced DSBs occur preferentially at binding sites of the most highly-transcriptionally active transcription factors in PC cells, including c-Myc, FOXA1, HOXB13, NKX3.1, and importantly, AR, resulting in target transcript downregulation. Further, RNA-seq reveals widespread miR-346 and shNORAD dysregulation of DNA damage, replication and cell cycle processes. NORAD drives target-directed miR decay (TDMD) of miR-346 as a novel genome protection mechanism: NORAD silencing increases mature miR-346 levels by several thousand-fold, and WT but not TDMD-mutant NORAD rescues miR-346-induced DNA damage. Importantly, miR-346 sensitises PC cells to DNA-damaging drugs including PARP inhibitor and chemotherapy, and induces tumour regression as a monotherapy in vivo, indicating that targeting miR-346:NORAD balance is a valid therapeutic strategy. CONCLUSIONS: A balancing act between miR-346 and NORAD regulates DNA damage and repair in PC. miR-346 may be particularly effective as a therapeutic in the context of decreased NORAD observed in advanced PC, and in transcriptionally-hyperactive cancer cells.

Thermographic visualization of facial vasoconstriction is associated with 30-day all-cause mortality in medical patients; prospective observational two-site cohort study.

INTRODUCTION: Quick and reliable assessment of acute patients is required for accurate triage. The temperature gradient between core and peripheral temperature could possibly instantly provide information on circulatory status. METHODS: Adult medical patients, who did not receive supplementary oxygen, attending two emergency departments, had a thermographic image taken on arrival. The association between 30-day mortality and gradients was tested using logistic regression. RESULTS: 726 patients were studied, median age was 64 years and 14 (1.9%) died within 30 days. There was a significant association between mortality and temperature gradient, comparable to vital signs, age, and clinical intuition. CONCLUSION: Temperature gradient between nose and eye had an acceptable discriminatory power for 30-day all-cause mortality.

Long-term efficacy and safety of biosimilar infliximab (CT-P13) after switching from originator infliximab: open-label extension of the NOR-SWITCH trial.

BACKGROUND AND OBJECTIVES: The 52-week, randomized, double-blind, noninferiority, government-funded NOR-SWITCH trial demonstrated that switching from infliximab originator to less expensive biosimilar CT-P13 was not inferior to continued treatment with infliximab originator. The NOR-SWITCH extension trial aimed to assess efficacy, safety and immunogenicity in patients on CT-P13 throughout the 78-week study period (maintenance group) versus patients switched to CT-P13 at week 52 (switch group). The primary outcome was disease worsening during follow-up based on disease-specific composite measures. METHODS: Patients were recruited from 24 Norwegian hospitals, 380 of 438 patients who completed the main study: 197 in the maintenance group and 183 in the switch group. In the full analysis set, 127 (33%) had Crohn's disease, 80 (21%) ulcerative colitis, 67 (18%) spondyloarthritis, 55 (15%) rheumatoid arthritis, 20 (5%) psoriatic arthritis and 31 (8%) chronic plaque psoriasis. RESULTS: Baseline characteristics were similar in the two groups at the time of switching (week 52). Disease worsening occurred in 32 (16.8%) patients in the maintenance group vs. 20 (11.6%) in the switch group (per-protocol set). Adjusted risk difference was 5.9% (95% CI -1.1 to 12.9). Frequency of adverse events, anti-drug antibodies, changes in generic disease variables and disease-specific composite measures were comparable between arms. The study was inadequately powered to detect noninferiority within individual diseases. CONCLUSION: The NOR-SWITCH extension showed no difference in safety and efficacy between patients who maintained CT-P13 and patients who switched from originator infliximab to CT-P13, supporting that switching from originator infliximab to CT-P13 is safe and efficacious.

Spatial and temporal changes in the morphology of preosteoblastic cells seeded on microstructured tantalum surfaces.

It has been widely reported that surface morphology on the micrometer scale affects cell function as well as cell shape. In this study, we have systematically compared the influence of 13 topographically micropatterned tantalum surfaces on the temporal development of morphology, including spreading, and length of preosteoblastic cells (MC3T3-E1). Cells were examined after 0.5, 1, 4, and 24 h on different Ta microstructures with vertical dimensions (heights) of 0.25 and 1.6 mum. Cell morphologies depended upon the underlying surface topography, and the length and spreading of cells varied as a function of time with regard to the two-dimensional pattern and vertical dimension of the structure. Microstructures of parallel grooves/ridges caused elongated cell growth after 1 and 4 h in comparison to a flat, nonstructured, reference surface. For microstructures consisting of pillars, cell spreading was found to depend on the distance between the pillars with one specific pillar structure exhibiting a decreased spreading combined with a radical change in morphology of the cells. Interestingly, this morphology on the particular pillar structure was associated with a markedly different distribution of the actin cytoskeleton. Our results provide a basis for further work toward topographical guiding of cell function.

Nanomechanical sensing of DNA sequences using piezoresistive cantilevers.

A microfabricated cantilever with an internal piezoresistive component has been sensitized with thiol tethered ss-DNA strands and utilized for an in situ, label-free, highly specific, and rapid DNA detection assay. The generation of a differential surface stress onto the functionalized cantilever surface upon target recognition has allowed nanomechanical identification of 12-nucleotide complementary DNA probes with single base mismatch discrimination (sensitivity of 0.2 microM). Interestingly, utilization of an overhang extension distal to the surface enhanced the sensitivity to the 0.01 microM level. The cantilever was functionalized by inkjet printing technology. Replacing the capture probe with locked nucleic acid (LNA) resulted in a faster target probe capture kinetics compared to DNA-DNA hybridization. The capabilities of the piezoresistive cantilever indicate future ergonomic convenience via miniaturization alternative to the conventional laser-based detection method for portable on-site applications.

Bioavailability of selenium from fish, yeast and selenate: a comparative study in humans using stable isotopes.

OBJECTIVE: To measure the bioavailability of selenium from cooked and raw fish in humans by estimating and comparing apparent absorption and retention of selenium in biosynthetically labelled fish with labelled selenate and biosynthetically labelled selenium in brewers yeast. DESIGN: The intervention study was a parallel, randomised, reference substance controlled design carried out at two different centres in Europe. SETTING: The human study was carried out at the Institute of Food Research, Norwich, UK and at TNO Nutrition and Food Research, Zeist, The Netherlands. SUBJECTS: In all, 35 male volunteers aged 18-50 y were recruited; 17 subjects were studied in Norwich (UK) and 18 in Zeist (Netherlands). All of the recruited subjects completed the study. INTERVENTIONS: Biosynthetically labelled trout fish (processed by two different methods), biosynthetically labelled brewers yeast and isotopically labelled selenate were used to estimate selenium apparent absorption and retention by quantitative analysis of stable isotope labels recovered in faeces and urine. Subjects consumed the labelled foods in four meals over two consecutive days and absorption was measured by the luminal disappearance method over 10 days. Urinary clearance of isotopic labels was measured over 7 days to enable retention to be calculated. RESULTS: Apparent absorption of selenium from fish was similar to selenate and there was no difference between the two processing methods used. However, retention of fish selenium was significantly higher than selenate (P<0.001). Apparent absorption and retention of yeast selenium was significantly different (P<0.001) from both fish selenium and selenate. CONCLUSION: Fish selenium is a highly bioavailable source of dietary selenium. Cooking did not affect selenium apparent absorption or retention from fish. Selenium from yeast is less bioavailable.

[Are obesity and overweight risk factors of asthma?]. / Er fedme og overvaegt risikofaktorer for asthma?

It has recently been suggested that overweight and obesity could be possible risk factors for asthma. A review of clinical and epidemiological studies of children and adults shows that a significant association between a high body mass index (BMI) and asthma can often be found. Yet, at present, we think that it is too early to conclude that overweight and obesity should be regarded as established risk factors for asthma. In particular, there is a need for more longitudinal studies to confirm the association between asthma and overweight and to shed light on which condition develops first. In addition, there is also a need for clinical studies of possible biological mechanisms that could link these two common conditions.

[Muscle tissue for diagnosis of neuromuscular diseases. Procedures of specimen taking, transport and preparation]. / Muskelvev for diagnostikk av neuromuskulaere sykdommer. Prosedyre for prøvetaking, transport og preparering.

In this review we describe surgical methods for open muscle biopsy. In order to obtain optimal tissue quality, correct handling of the muscle specimens at all stages of the procedure is essential. A technologist who is familiar with the procedure should be present in the operating room. This is important to ensure that specimens for histochemical or biochemical studies are frozen immediately and specimens for electron microscopy are fixed without delay. We describe procedures for processing the specimens for cryostat sections, paraffin sections and electron microscopy, and a simple method for handling muscle specimens in local hospitals before transfer to a hospital with a neuropathological service.

Decreased concentration of hemoglobin, accumulation of lipid oxidation products and unchanged skeletal muscle in Atlantic salmon (Salmo salar) fed low dietary vitamin E.

Atlantic salmon (Salmo salar) fry, initial weight 0.16 g, were fed a semipurified diet with 0, 15, 30, 60 or 120 mg dl-α-tocopheryl acetate/kg. After 24 weeks, the first two of these groups were extinct, and the fish receiving 30 mg/kg were clearly vitamin E deficient. Vitamin E deficient fish had low hemoglobin levels, characterized by a combination of reduced cellular hemoglobin concentration, red cell volume and red cell number, and an increased number and fraction of immature red blood cells. The hemoglobin concentration decreased over the decreasing range of experimental dl-ga-tocopheryl acetate levels. Therefore, even if 60 mg dl-α-tocopheryl acetate/kg gave good survival, this level was clearly physiologically suboptimal. Ceroid accumulated in the liver of fish fed 30 mg vitamin E/kg, and autofluorescent inclusions were found in the red blood cells of fish fed 30 and 60 mg vitamin E/kg. Degeneration of skeletal muscle was not observed in the present study.

Xanthogranulomatous cholecystitis. A clinicopathological study of 20 cases and review of the literature.

Xanthogranulomatous cholecystitis (XGC) is a focal or diffuse destructive inflammatory process of the gall bladder, characterized macroscopically by yellowish tumour-like masses in the wall of the gall bladder. Microscopically, it is characterized in the early stages by a large number of foamy histiocytes and acute inflammatory cells. Later stages demonstrate increasing fibrosis. The gall bladder from 20 of 352 consecutive patients subjected to cholecystectomy showed XGC. Gall stones were found in the gall bladder of all 20 patients and in the ductus choledochus in 3 cases. Perforation of the gall bladder was observed at operation in six cases; in one case there was also a fistula to the colon. A perivesical abscess was found in five other cases. Adhesions to the surrounding structures were seen in a total of 16 cases. Pathogenetically, XGC is probably due to an interplay between obstruction of the gall flow, infection with subsequent inflammation, and leakage of gall fluid to the tissue, where histiocytes accumulate and phagocytize the bile pigment, haemosiderin and cholesterol, resulting in the formation of xanthoma cells. The correct diagnosis of XGC is important for several reasons, first and foremost due to the high frequency of complications, but not least because the condition may give rise peroperatively to the suspicion of malignancy. The new laparoscopic method for cholecystectomies further stresses the necessity of correct preoperative diagnosis of complicating disease.

Phaeochromocytoma--an unusual cause of haematuria.

A patient was admitted to hospital because of haematuria. A papillomatous bladder tumor was removed. Histological examination revealed a phaeochromocytoma. Fourteen days postoperatively the urinary excretion rates of catecholamines were elevated but normalized afterwards. Abdominal computed tomography and metaiodobenzylguanidine scintigraphy were normal. No hypertension or characteristic micturitional attacks preceded removal of the tumor.

Anorectal dysfunction in patients with urologic disturbance due to multiple sclerosis.

Anorectal function was evaluated in 11 patients with voiding dysfunction due to multiple sclerosis. In six patients with constipation, three also had symptoms of obstructed defecation and one patient was incontinent due to stercoral diarrhea. One patient was only fecal incontinent and one patient had obstructed defecation as the only symptom. Three patients had no anorectal symptoms. Anal manometry in the women compared with a control group revealed significant lower anal resting and squeeze pressures, although no significant difference of rectal sensation to distention with air was found. Pudendal nerve terminal latencies were obtained in seven patients and were all normal. In four patients latency could not be demonstrated due to poor contraction of the sphincter on stimulation of the pudendal nerve. Two of these patients were incontinent and two had both constipation and obstructed defecation. It is concluded that patients with voiding symptoms due to multiple sclerosis often reveal anorectal symptoms or motility disorders. Although anal sphincter function is reduced, fecal incontinence is not prevalent in this group. The reason for this lies probably in the fact that many of the patients are constipated, thus securing fecal continence.

Influence of peptides on anorectal function.

Immunohistochemical and immunochemical studies on biopsies from the human rectum and anal canal have shown several regulatory peptides present in the muscle layers and the mucosa, suggesting a regulatory action on defecation. This view has been supported by studies of anorectal function in man during administration of different peptides. The physiological implications of these observations remain obscure.

Implantation of artificial sphincter for anal incontinence. Report of five cases.

The implantation of an artificial anal sphincter in five patients with anal incontinence of neuromuscular origin is described. The prosthesis, an AMS 800 artificial urinary sphincter (American Medical Systems), worked well with solid or semisolid stool, but less satisfactorily when diarrhea was present. Manometric and defecographic studies seem to indicate that the effect of the prosthesis may be ascribed partly to maintenance of an acute anorectal angle even during straining.

Influence of gastrointestinal neuropeptides on the anal canal.

To evaluate the possible effect of gastrointestinal neuropeptides on anal function, the effect of somatostatin, enkephalin, VIP, and substance P on anal canal pressure and electromyographic response of the external anal sphincter was studied in healthy subjects. Enkephalin and somatostatin elicited a significant decrease in anal canal pressure after a bolus injection of 1 microgram/kg body weight whereas VIP and substance P had no effect. Future studies must show whether these effects are of pharmacologic importance and if these peptides participate in the physiologic regulation of anorectal function.

Xanthogranulomatous inflammation of the female genital tract.

We describe three cases of xanthogranulomatous inflammation in the female genital tract--one affecting endometrium, tube and ovary, one affecting tube, ovary and parametrium and one confined to the endometrium. To date, xanthogranulomatous inflammation in the female genital tract has been reported in a total of 19 cases including the present three. The inflammation most often affects the endometrium but involvement of the vagina, cervix, fallopian tube and ovary may also occur.

Implantation of artificial sphincter for anal incontinence.

The implantation of an artificial anal sphincter in a man with severe anal incontinence an myasthenia gravis is described. The prosthesis used, an "AMS 800' artificial urinary sphincter, gave the patient complete control of defecation. The technique offers the possibility of curing anal incontinence of neuromuscular origin, for which there has been no treatment until know.

Effect of sham-feeding on glucose tolerance and insulin secretion.

Glucose, 25 g, was infused iv with or without sham-feeding in seven normal males. Sham-feeding improved glucose tolerance, incremental area of blood glucose being 63% (P less than 0.05) of that during iv glucose without sham-feeding. The actual insulin secretion evaluated from the total area under the C-peptide and insulin curves did not differ during iv glucose with or without sham-feeding. These results suggest that the cephalic-vagal reflex improves glucose tolerance during iv glucose, independent of changes in beta-cell function.