Sarcopenia in critically ill children: A bedside assessment using point-of-care ultrasound and anthropometry.

BACKGROUND & AIMS: Due to the lack of validated methods of muscle assessment, sarcopenia is not well described in critically ill children. The main objectives of this study were to assess muscle wasting using point-of-care ultrasound (POCUS) and anthropometry, as well as its association with nutrition delivery in PICU. METHODS: This was a single-center, prospective cohort study, including consecutive children admitted to the PICU. Quadriceps femoris muscle thickness (QFMT) and anthropometrics measurements were performed at admission and then weekly until the 14th day of the PICU stay. The three moments of assessment were defined as T0 (baseline), T1 (7th day) and T2 (14th day). For analysis purposes, participants assessed only in T0 and T1 were defined as Subgroup 1, while those assessed in T0, T1 and T2 were defined as Subgroup 2. Actual total daily intake was determined by patient intake records until discharge or during the first 14 full days of PICU admission. RESULTS: In all, 119 patients were included with a median age of 12.0 months (IQR 4.0-42.5). In Subgroup 1, QFMT significantly decreased between T0 and T1 (-12.93 ± 14.07 %; p < 0.001), and the same was observed in Subgroup 2 (-13.81 ± 13.05 %; p < 0.001). However, no differences in QFMT was observed between T1 and T2 (-2.06 ± 13.80 %; p = 0.936). Triceps skinfold thickness, mid-upper arm circumference, and upper arm muscle area presented a similar pattern of changes between periods in both groups. Decrease of QFMT at T1 was significantly correlated with the cumulative protein deficit in both subgroups, but not with the cumulative energy deficit. CONCLUSION: Substantial muscle wasting occurs early in critically ill children and may be related to insufficient protein delivery. Anthropometric measurements are valuable in PICU and POCUS has the potential to play a major role in sarcopenia assessment during critical illnesses. TRIAL REGISTRATION: Brazilian Clinical Trials registry, registration number: RBR-85YYGN.

Contamination of single-use bronchoscopes in critically ill patients.

Disposable bronchoscopes such as the Ambu aScopeTM 3 are marketed as 'single use' The risks of contamination from prolonged device storage before possible re-use are unknown. Following clinical bronchoscopy in patients whose lungs were mechanically ventilated, 20 aScopeTM 3's bronchoscopes received a standard 'social clean' and were then stored. Subsequent paired saline flush and swab samples were taken at time zero, and at 24 h and 48 h. Positive microbiological cultures were obtained from at least one time point from 16 of the 20 bronchoscopes. Pathogens considered at high risk of causing pneumonia were isolated from seven bronchoscopes, with significant quantities from six of them. Our study demonstrates that aScopeTM 3's should not be re-used on the same patient, as clinically significant growth of micro-organisms occurs frequently, despite adequate social cleaning. Culture of bronchoscopes themselves may be a potentially useful diagnostic tool in the context of pulmonary infection. Our data make it clear that these devices are single use and not single patient use.

Multilocus phylogeographic assessment of the California Mountain Kingsnake (Lampropeltis zonata) suggests alternative patterns of diversification for the California Floristic Province.

Phylogeographic inference can determine the timing of population divergence, historical demographic processes, patterns of migration, and when extended to multiple species, the history of communities. Single-locus analyses can mislead interpretations of the evolutionary history of taxa and comparative analyses. It is therefore important to revisit previous single-locus phylogeographic studies, particularly those that have been used to propose general patterns for regional biotas and the processes responsible for generating inferred patterns. Here, we employ a multilocus statistical approach to re-examine the phylogeography of Lampropeltis zonata. Using nonparametic and Bayesian species delimitation, we determined that there are two well-supported species within L. zonata. Ecological niche modelling supports the delimitation of these taxa, suggesting that the two species inhabit distinct climatic environments. Gene flow between the two taxa is low and appears to occur unidirectionally. Further, our data suggest that gene flow was mediated by females, a rare pattern in snakes. In contrast to previous analyses, we determined that the divergence between the two lineages occurred in the late Pliocene (c. 2.07 Ma). Spatially and temporally, the divergence of these lineages is associated with the inundation of central California by the Monterey Bay. The effective population sizes of the two species appear to have been unaffected by Pleistocene glaciation. Our increased sampling of loci for L. zonata, combined with previously published multilocus analyses of other sympatric species, suggests that previous conclusions reached by comparative phylogeographic studies conducted within the California Floristic Province should be reassessed.

Phylogenetic relationships of the Cretaceous frog Beelzebufo from Madagascar and the placement of fossil constraints based on temporal and phylogenetic evidence.

The placement of fossil calibrations is ideally based on the phylogenetic analysis of extinct taxa. Another source of information is the temporal variance for a given clade implied by a particular constraint when combined with other, well-supported calibrations. For example, the frog Beelzebufo ampinga from the Cretaceous of Madagascar has been hypothesized to be a crown-group member of the New World subfamily Ceratophryinae, which would support a Late Cretaceous connection with South America. However, phylogenetic analyses and molecular divergence time estimates based on other fossils do not support this placement. We derive a metric, Δt, to quantify temporal divergence among chronograms and find that errors resulting from mis-specified calibrations are localized when additional nodes throughout the tree are properly calibrated. The use of temporal information from molecular data can further assist in testing phylogenetic hypotheses regarding the placement of extinct taxa.

A violence-prevention and evaluation project with ethnically diverse populations.

The purpose of this paper is to describe some of the challenges encountered and lessons learned while providing and evaluating a violence-prevention program for and with ethnically diverse populations in child care settings. The paper discusses Safe Start, a violence prevention education program for child care staff and parents, and the evaluation of the program. Safe Start was designed to include culturally relevant content to increase cultural awareness for child care staff and parents from diverse ethnic backgrounds. The evaluation project enrolled child care centers with families representing the ethnically diverse communities in which they were located. Violence prevention research involving children from diverse ethnic backgrounds presents new methodologic challenges, but also provides new opportunities for creative, novel methods. This paper describes some of the challenges encountered with curriculum development, staff recruitment, instrument selection, and data collection procedures.

Isolation, structures, and biologic activity of neurotensin-related peptides generated in extracts of avian tissue.

Two immunoreactive neurotensin-related peptides generated by the action of endogenous protease(s) on protein substrates during acid extraction of avian tissues have been isolated from extracts of turkey skin and proventriculus. One was identified as the pentadecapeptide, H-Phe-Glu-Arg-Phe-Gln-Gly-Met-Arg-Thy-Arg-Gly-Pro-Tyr-Phe-Leu-OH and the other was its C-terminal octapeptide fragment. Each peptide showed partial homology to the C-terminal, biologically active region of avian neurotensin, and isolated preparations displayed pharmacologic activity at submicromolar concentrations. Synthetic preparations were shown to be indistinguishable from the native peptides during high pressure liquid chromatography (HPLC) and bioassay. Analysis by HPLC indicated that similar peptides could be generated in extracts of proventriculus, pancreas, small intestine, skin, heart, lung, and skeletal muscle. These results, establishing the presence of a neurotensin-related sequence which can be liberated from protein(s) by the action of tissue enzyme(s), suggest that peptide(s) similar to neurotensin may be rapidly formed in order to promote physiologic regulation in multiple tissue(s).

Radioimmunoassay for Lys8, Asn9, neurotensin 8-13: tissue and subcellular distribution of immunoreactivity in chickens.

A sensitive and specific radioimmunoassay (RIA) for Lys8, Asn9, neurotensin 8-13 (LANT-6) has been developed which utilizes 125I-labeled LANT-6 and rabbit antisera raised towards conjugates of synthetic LANT-6 and bovine thyroglobulin. The antiserum described (TG-22) allows the detection of ca 100 fmol of LANT-6 and crossreacts less than 0.01% with chicken or bovine NT. Dose-response relationships for the native (chicken) and synthetic peptides were indistinguishable. Using this assay the distribution of immunoreactive LANT-6 (iLANT-6) through various tissues of the chicken was studied and compared with that of chicken NT (iNT) determined by RIA. Both iNT and ILANT-6 were found primarily in the brain and gastrointestinal tract, however, their regional distributions were found to differ. Subcellular distribution studies in homogenates of chicken brain indicated that both iNT and iLANT-6 were associated with synaptosome-like and vesicle-like particles. In homogenates of small intestine, pancreas and colon iNT and iLANT-6 appeared to be within osmotically sensitive, sedimentable particles. Analyses using high pressure liquid chromatography established that chicken iLANT-6 co-eluted with the synthetic peptide and that similar substances were present in extracts of rat brain and intestine. These results are consistent with "messenger' roles for these peptides.

Amphibian neurotensin (NT) is not xenopsin (XP): dual presence of NT-like and XP-like peptides in various amphibia.

To clarify whether xenopsin (XP) is the amphibian counterpart of mammalian neurotensin (NT), extracts of skin, brain, and intestine from representative amphibians were subjected to immunochemical, chromatographic, and biological analyses. The results indicated the dual presence of NT- and XP-like peptides in extracts of tissues from Xenopus laevis, Rana catesbeiana, Rana pipiens, Bufo marinus, Bufo americanus, and Necturus maculosus, which were separated during gel chromatography on Sephadex G-25 and high pressure liquid chromatography on mu-Bondapak C-18. Immunochemical studies, employing three different region-specific antisera toward NT (ox and man) and one antiserum towards XP (Xenopus laevis), indicated that the NT-like peptides shared COOH-terminal homologies with NT and differed at their NH2-termini. TWo classes of NT-like peptides could be distinguished on the basis of their distributions in tissues and their cross-reactivities with the antisera; immunoreactive NT measured using antiserum HC-8 tended to be found primarily in brain and intestine, whereas that reactive with antiserum PGL-4 was most concentrated in stomach, liver, and pancreas. Although also present in brain and intestine, immunoreactive XP was highest in stomach, pancreas, and skin. Partially purified immunoreactive NT and XP obtained from gastrointestinal tissues of Xenopus laevis and Bufo marinus were shown to increase the hematocrit and induce cyanosis in anesthesized rats. These findings indicate the presence of both NT- and XP-like peptides in neural and gastrointestinal tissues from several amphibia and suggest the possibility that XP-like peptides (apart from NT) may exist in other animals.

Distribution and immunochemical character of neurotensin-like material in representative vertebrates and invertebrates: apparent conservation of the COOH-terminal region during evolution.

Using a radioimmunoassay for bovine neurotensin (NT) and various region specific antisera which react selectively with different portions of the molecule, the presence of immunoreactive NT (iNT) in a wide range of vertebrate and invertebrate species has been demonstrated. While antisera directed towards the N-terminal region of NT recognized only mammalian forms of NT, antisera directed towards the C-terminal region of NT recognized materials from all species examined, including representatives of all vertebrates and invertebrate classes. When extracts of the brain and gut of the vertebrates examined were chromatographed on Sephadex G-25 multiple NT-like substances were observed, and the patterns of iNT obtained seemed to fall into three groups: (a) mammals, (b) birds and reptiles and (c) amphibians and fish. Extracts of invertebrates also exhibited multiple peaks of iNT on Sephadex G-25 and the profiles observed resembled those for lower vertebrates. Partially purified iNT obtained from chicken, turtle, dogfish and lobster was shown to increase hematocrit and induce cyanosis in anesthetized rats. These findings indicate (a) that NT-like substances appear to be present throughout the animal kingdom, (b) that the C-terminal region of NT is highly conserved while the N-terminal region varies, and (c) that in any one animal multiple substances sharing C-terminal homologies with NT exist. These findings are consistent with the notion that NT and related peptides participate in important processes basic to animal life and that their functioning depends highly upon elements located in their C-terminal regions. They further suggest the existence of an entire family of NT-related peptides in each animal form, possibly distributed differently and functioning differently in the various organs of the animal.