Dupilumab for chronic rhinosinusitis with nasal polyps: real-life retrospective 12-month effectiveness data.

BACKGROUND: Dupilumab, an IL-4/13 receptor inhibitor, is approved for the treatment of uncontrolled chronic rhinosinusitis with nasal polyps (CRSwNP). METHODOLOGY: We evaluated the effectiveness and safety of dupilumab for CRSwNP based on retrospective 12-month follow-up data of 41 patients. We analysed nasal endoscopy scores, patient-reported outcome measures (PROMs), 12-item SniffinSticks odor identification test (SSIT-12), total serum IgE, serum Eosinophilic Cationic Protein (ECP), and total blood eosinophil count (BEC). We performed statistical analysis using non-parametric ANOVA-type models and Spearman's correlation. RESULTS: At month 1, endoscopy scores, PROMs and SSIT-12 showed meaningful improvements that were maintained until month 12. Initial elevations in both median ECP and BECs returned to near baseline levels by month 12. The percentage of patients withBEC ≤ 0.6 remained increased at month 12 (42.1%) compared to baseline (19.5%). Total serum IgE levels decreased progressively and correlated with nasal polyp scores at month 12. "Adequate response" was reached in 86.8% of our cohort. CONCLUSIONS: Our data suggest that dupilumab is effective for the treatment of CRSwNP. The potential for short- and long-term BEC elevations in some CRSwNP patients should be carefully monitored.

Tracking of bone mass from childhood to puberty: a 7-year follow-up. The CHAMPS study DK.

Bone mass in childhood is highly influenced by puberty. At the same age, bone mass was higher for pubertal than pre-pubertal children. A high level of tracking during 7 years from childhood through puberty was shown, indicating that early levels of bone mass may be important for later bone health. INTRODUCTION: Bone mass development in childhood varies by sex and age, but also by pubertal stage. The objectives of this study were to (1) describe bone mass development in childhood as it relates to pubertal onset and to (2) determine the degree of tracking from childhood to adolescence. METHODS: A longitudinal study with 7 years of follow-up was initiated in 2008 to include 831 children (407 boys) aged 8 to 17 years. Participants underwent whole body dual-energy X-ray absorptiometry (DXA) scanning, blood collection to quantify luteinizing hormone levels, and Tanner stage self-assessment three times during the 7-year follow-up. Total body less head bone mineral content, areal bone mineral density, and bone area were used to describe development in bone accrual and to examine tracking over 7 years. RESULTS: Bone mass in pubertal children is higher than that of pre-pubertal children at the same age. Analysing tracking with quintiles of bone mass Z-scores in 2008 and 2015 showed that more than 80% of participants remained in the same or neighbouring quintile over the study period. Tracking was confirmed by correlation coefficients between Z-scores at baseline and 7-year follow-up (range, 0.80-0.84). CONCLUSIONS: Bone mass is highly influenced by pubertal onset, and pubertal stage should be considered when examining children's bone health. Because bone mass indices track from childhood into puberty, children with low bone mass may be at risk of developing osteoporosis later in life.

Functional regeneration of intraspinal connections in a new in vitro model.

Regeneration in the adult mammalian spinal cord is limited due to intrinsic properties of mature neurons and a hostile environment, mainly provided by central nervous system myelin and reactive astrocytes. Recent results indicate that propriospinal connections are a promising target for intervention to improve functional recovery. To study this functional regeneration in vitro we developed a model consisting of two organotypic spinal cord slices placed adjacently on multi-electrode arrays. The electrodes allow us to record the spontaneously occurring neuronal activity, which is often organized in network bursts. Within a few days in vitro (DIV), these bursts become synchronized between the two slices due to the formation of axonal connections. We cut them with a scalpel at different time points in vitro and record the neuronal activity 3 weeks later. The functional recovery ability was assessed by calculating the percentage of synchronized bursts between the two slices. We found that cultures lesioned at a young age (7-9 DIV) retained the high regeneration ability of embryonic tissue. However, cultures lesioned at older ages (>19 DIV) displayed a distinct reduction of synchronized activity. This reduction was not accompanied by an inability for axons to cross the lesion site. We show that functional regeneration in these old cultures can be improved by increasing the intracellular cAMP level with Rolipram or by placing a young slice next to an old one directly after the lesion. We conclude that co-cultures of two spinal cord slices are an appropriate model to study functional regeneration of intraspinal connections.

Prospective association of adiposity and cardiorespiratory fitness with cardiovascular risk factors in healthy children.

This study aimed to examine the prospective association of three different measures of adiposity and cardiorespiratory fitness (CRF) with 2-year change in cardiovascular disease (CVD) risk factors in children. Two-year longitudinal data in schoolchildren aged 7-11 years (n = 365-729) was used. Total body fat (TBF) from dual-energy x-ray absorptiometry, body mass index (BMI), waist circumference (WC), CRF, blood samples, and blood pressure were obtained in 2008 and 2010 in the Childhood Health, Activity, and Motor Performance School Study in Denmark (CHAMPS study-DK). Greater adiposity at baseline was associated with increased CVD risk factor levels at follow-up. The magnitudes of associations were similar regardless of adiposity measure [TBF%: ß 0.30, 95% confidence interval (CI): 0.21-0.39; BMI: ß 0.24, 95% CI: 0.14-0.33; WC: ß 0.20, 95% CI: 0.10-0.31], and no evidence of nonlinear relationships was observed. We found less strong associations of CRF with increase in CVD risk factor levels after adjusting for adiposity; however, increasing CRF was still favorably associated with decrease in CVD risk factor levels among boys. Results suggest that any effort to shift the population distribution of adiposity downward would be valuable for early CVD prevention. The association of CRF with CVD risk factors was largely explained by adiposity, particularly among girls.

Functional nuclear organization of transcription and DNA replication: a topographical marriage between chromatin domains and the interchromatin compartment.

We studied the nuclear topography of RNA transcription and DNA replication in mammalian cell types with super-resolution fluorescence microscopy, which offers a resolution beyond the classical Abbe/Raleigh limit. Three-dimensional structured illumination microscopy (3D-SIM) demonstrated a network of channels and wider lacunas, called the interchromatin compartment (IC). The IC starts at nuclear pores and expands throughout the nuclear space. It is demarcated from the compact interior of higher-order chromatin domains (CDs) by a 100-200-nm thick layer of decondensed chromatin, termed the perichromatin region (PR). Nascent DNA, nascent RNA, RNA polymerase II (RNA Pol II), as well as histone modifications for transcriptionally competent/active chromatin, are highly enriched in the PR, whereas splicing speckles are observed in the interior of the IC. In line with previous electron microscopic evidence, spectral precision distance/position determination microscopy (SPDM) confirmed the presence of RNA Pol II clusters indicative of transcription factories. Still, a substantial part of transcription apparently takes place outside of such factories. Previous electron microscopic evidence has suggested that the functional nuclear organization of DNA replication depends on brownian movements of chromatin between the CD interior and the PR. As an incentive for future studies, we hypothesize that such movements also take place during transcription, i.e., only the actually transcribed part of a gene may be located within the PR, whereas its major part, including previously or later transcribed sequences, is embedded in a higher-order chromatin configuration in the interior of the CD.

Does rapid intraoperative parathyroid hormone analysis predict cure in patients undergoing surgery for primary hyperparathyroidism? A prospective study.

BACKGROUND AND AIMS: Preoperative 99mTc-sestamibi scintigraphy is used by many surgeons to identify the anatomical location of pathological parathyroid glands in patients undergoing surgical treatment for hyperparathyroidism. However, false negative results do occur. It has been suggested that intraoperative parathyroid hormone (PTH) analysis may enhance the possibility of performing successful focused, unilateral neck surgery in these patients. This study aimed to evaluate whether an adequate fall in intraoperative parathyroid hormone values predicts the removal of all hyperfunctioning parathyroid tissue and postoperative normocalcemia. MATERIAL AND METHODS: One hundred consecutive patients undergoing surgery for hyperparathyroidism had preoperative 99mTc-sestamibi scintigraphy and intraoperative parathyroid hormone (PTH) analysis. A fall in intraoperative PTH value by more than 50% of baseline value ended the procedure. This prospective study presents the clinical and biochemical results. RESULTS: The overall sensitivity of the 99mTc-sestamib scintigraphy was 88% and for single adenomas 95%. The scintigraphy failed to detect the correct pathology in all cases with multiglandular disease (7 patients). A fall in intraoperative PTH value by more than 50% of baseline value was achieved in all patients. The combination of intraoperative PTH analysis and 99mTc-sestamibi scintigraphy enabled us to limit the operation to a focused, unilateral operation in 87 of the 100 patients. All patients were normocalcemic postoperatively. CONCLUSIONS: A fall in intraoperative PTH value more than 50 % of baseline value seems to predict postoperative normocalcemia and the removal of all hyperfunctioning parathyroid tissue. Bilateral neck exploration is avoided in the majority of patients.

Nerve involvement in fluid transport in the inflamed rat jejunum.

Net fluid transport was measured in denervated jejunal segments of rats infected with larvae of Nippostrongylus brasiliensis. On days 6-9 after nematode inoculation, when the jejunal segment exhibited macroscopic and microscopic signs of inflammation, net fluid absorption was noticeably attenuated compared with control, and in eight of 26 experiments a net fluid secretion was seen. To determine whether enteric nerves participated in the response, intravenous hexamethonium (10 mg/kg body weight) was given or lidocaine (1% solution) was placed on the serosa of the intestinal segment. Both drugs significantly reduced fluid secretion or increased fluid absorption. The effect was more pronounced the lower the rate of fluid absorption or the higher the rate of fluid secretion. The inflammatory response influenced intestinal fluid transport partly via activation of the enteric nervous system. It was estimated that 50-60% of the change in fluid transport caused by the parasite could be ascribed to activation of intramural nervous reflexes. The effect of hexamethonium indicates that a cholinergic synapse is present in the secretory nervous reflux activated by inflammation. Experiments were also performed on animals on days 11-14 after infection when the nematodes had been expelled from the animal. A large net fluid absorption was then recorded.

An improved procedure for purifying 5'-nucleotidase from various sources. Evidence for tissue and species differences in their molecular mass and affinity for F-actin.

5'-Nucleotidase from chicken gizzard smooth muscle has been extracted, using a sulfobetaine derivate of cholic acid, and purified to homogeneity by employing three chromatographic steps. It is shown that the purification scheme can be applied to 5'-nucleotidase from other sources, such as rat liver. On sodium dodecyl sulfate polyacrylamide gels, stained with silver nitrate, the purified enzyme from chicken gizzard shows a single polypeptide band with an apparent molecular mass of 79 kDa. The enzyme purified from rat liver exhibits a molecular mass of 73 kDa in agreement with published data [Bailyes, E.M., Soos, M., Jackson, P., Newby, A. C., Siddle, K. & Luzio, J.P. (1984) Biochem. J. 221, 369-377). Gel filtration, using non-denaturating detergent solutions, indicates that the native enzyme may exist as a homodimer (152 kDa) or homotetramer (310 kDa). Antibodies raised against the enzyme purified from chicken gizzard bind only 5'-nucleotidase, solubilized from chicken muscular sources, when immobilized, but not from chicken or rat liver. The existence of tissue specific variants of 5'-nucleotidase is therefore postulated and it appears that these particular isoforms can also be classified in membranous and secretory forms of 5'-nucleotidase. They also differ in their mode of interaction with actin. The AMPase activity of the membranous (= muscular) isoform is inhibited to a considerably higher percentage by F-actin than the enzyme isolated from rat liver.

Isolation of 5'-nucleotidase from chicken gizzard, its properties, and subcellular localization in chicken tissues using monospecific antibodies.

After solubilization with zwitterionic detergents 5'-nucleotidase was purified to homogeneity from chicken gizzard. Purified 5'-nucleotidase appeared to be composed of a single polypeptide chain of 79 kDa as revealed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE); gel filtration studies in the presence of detergents, however, indicated that the native enzyme is a homodimer. Antisera against purified chicken gizzard 5'-nucleotidase were raised in rabbits, they were shown to inhibit the enzymic activity of different 5'-nucleotidase preparations in a species and tissue specific manner. On frozen sections the antisera stained the cell periphery of chicken gizzard smooth and skeletal muscle and faintly stained cardiac muscle cells when using the indirect immunofluorescent technique. In chicken cardiac tissue, a prominent staining of the vascular system also became apparent. In avian and rat non-muscle tissues (hepatic and pancreatic tissue) the vascular system was always found to be brightly stained, i.e. the vascular smooth muscle and endothelial cells. On frozen sections of chicken liver the sinusoidal region of the hepatocytes was brightly stained, the bile canalicular region, however, only faintly. Using the immunocytochemical technique, a more prominent tissue specificity rather than species specificity of the available antisera became apparent. This may therefore reflect the existence of tissue-specific isoforms of the enzyme.

Fine structure of the forelimb regenerate of the African clawed toad, Xenopus laevis.

Forelimb regenerates from postmetamorphic Xenopus froglets were examined at various stages postamputation by light microscopy and scanning and transmission electron microscopy. The outside surface of the wound epithelium was found to exhibit progressive changes following amputation with a distinct difference in appearance between stump epidermis and wound epithelium at all stages examined. The internal structure of the wound epithelium is characterized by loosely arranged cells with numerous cell junctions and abundant intracellular filaments. The wound epithelium is separated at an early stage from the underlying cells by a thick band of extracellular matrix. Cells accumulating beneath the wound epithelium were morphologically similar to blastemal cells from completely regenerating limbs in other species but no evidence of myogenesis or abortive myogenesis was seen. Blastemal cells from the central portion of the regenerate were observed at varius stages of chrondrogenesis with those immediately beneath the wound epithelium least advanced in this respect. Those located more laterally appear not to be directly involved in chondrogenesis. Although the usual explanation for lack of complete regeneration in this species is inadequate innervation of the regenerate, the fine structure of the regenerating spikes noted here is markedly different than that of denervated, amputated newt limbs.

Quantitative behavioral studies of misdirected responses in skin-grafted Rana pipiens.

The experiments reported here describe quantitatively wiping responses elicited from three equal body sections of adult Rana pipiens having had one of two types of skin-graft operations as tadpoles. These experiments assessed the development of aberrant wiping responses in individuals and populations of grafted animals. Grafting parameters critical for the development of aberrant wiping responses were also determined. These studies indicated that the phenomenon of "misdirected responses" and the extent of these responses is highly variable among grafted individuals and between different populations of grafted animals. In addition, the development of "misdirected responses" is independent of premetamorphic stimulation and of a critical grafting period. The proportion of "misdirected responses" elicited upon cutaneous stimulation of individuals appears to be stable and predictable after metamorphosis. "Misdirected responses" could be elicited from animals with grafts including over 40% of the trunk skin. Wiping responses and grafting parameters of skin-grafted R. pipiens in other studies are also presented and possible sources of interstudy variation discussed.

Neurophysiological and behavioral evidence for selective reinnervation in skin-grafted Rana pipiens.

Rana pipiens with skin dorosoventrally reversed can respond to stimulation of the back with forelimb wipes to the belly and to stimulation of the belly with hindlimb wipes to the back. These "misdirected wiping responses" have been explained in terms of two alternative hypotheses of nerve regeneration: nerve respecification or selective reinnervation. Experimental behavioral and neurophysiological experiments reported here support the selective reinnervation hypothesis. Severing ventral nerves, which normally innervate the belly, greatly reduced the percentage of misdirected responses on stimulation of belly skin grafted to the back, while severing dorsal nerves, which normally innervate the back, increased the percentage of misdirected responses elicited under the same circumstances. Moreover, neurophysiological recordings of grafted animals showed three effects of skin grafting on nerve distributions: (i) termination of dorsal and ventral nerve receptive field at graft edges; (ii) overlap of nonadjacent ventral nerve receptive fields; and (iii) dorsal coursing of ventral nerves to reinnervate target belly skin displaced to the back. These neurophysiological observations, and particularly the third effect, also support selective reinnervation as the mechanism of nerve regeneration in skin-grafted Rana pipiens.