The effects of sugammadex vs. neostigmine on postoperative respiratory complications and advanced healthcare utilisation: a multicentre retrospective cohort study.

Reversing neuromuscular blockade with sugammadex can eliminate residual paralysis, which has been associated with postoperative respiratory complications. There are equivocal data on whether sugammadex reduces these when compared with neostigmine. We investigated the association of the choice of reversal drug with postoperative respiratory complications and advanced healthcare utilisation. We included adult patients who underwent surgery and received general anaesthesia with sugammadex or neostigmine reversal at two academic healthcare networks between January 2016 and June 2021. The primary outcome was postoperative respiratory complications, defined as post-extubation oxygen saturation < 90%, respiratory failure requiring non-invasive ventilation, or tracheal re-intubation within 7 days. Our main secondary outcome was advanced healthcare utilisation, a composite outcome including: 7-day unplanned intensive care unit admission; 30-day hospital readmission; or non-home discharge. In total, 5746 (6.9%) of 83,250 included patients experienced postoperative respiratory complications. This was not associated with the reversal drug (adjusted OR (95%CI) 1.01 (0.94-1.08); p = 0.76). After excluding patients admitted from skilled nursing facilities, 8372 (10.5%) patients required advanced healthcare utilisation, which was not associated with the choice of reversal (adjusted OR (95%CI) 0.95 (0.89-1.01); p = 0.11). Equivalence testing supported an equivalent effect size of sugammadex and neostigmine on both outcomes, and neostigmine was non-inferior to sugammadex with regard to postoperative respiratory complications or advanced healthcare utilisation. Finally, there was no association between the reversal drug and major adverse cardiovascular events (adjusted OR 1.07 (0.94-1.21); p = 0.32). Compared with neostigmine, reversal of neuromuscular blockade with sugammadex was not associated with a reduction in postoperative respiratory complications or post-procedural advanced healthcare utilisation.

Quantification of HSV-1-mediated expression of the ferritin MRI reporter in the mouse brain.

The development of effective strategies for gene therapy has been hampered by difficulties verifying transgene delivery in vivo and quantifying gene expression non-invasively. Magnetic resonance imaging (MRI) offers high spatial resolution and three-dimensional views, without tissue depth limitations. The iron-storage protein ferritin is a prototype MRI gene reporter. Ferritin forms a paramagnetic ferrihydrite core that can be detected by MRI via its effect on the local magnetic field experienced by water protons. In an effort to better characterize the ferritin reporter for central nervous system applications, we expressed ferritin in the mouse brain in vivo using a neurotropic herpes simplex virus type 1 (HSV-1). We computed three-dimensional maps of MRI transverse relaxation rates in the mouse brain with ascending doses of ferritin-expressing HSV-1. We established that the transverse relaxation rates correlate significantly to the number of inoculated infectious particles. Our results are potentially useful for quantitatively assessing limitations of ferritin reporters for gene therapy applications.

Single chain ferritin chimera as an improved MRI gene reporter.

Imaging gene expression non-invasively and deep into opaque tissues has been a long-standing goal of molecular science. Optical gene reporters such as green fluorescent protein and luciferase have revolutionized cellular and molecular biology, however their in vivo application is limited, due to poor tissue penetration of visible light. The iron storage protein ferritin forms a paramagnetic ferrihydrite core that affects the relaxation rate of surrounding nuclear spins. Ferritin has recently emerged as an MRI gene reporter for molecular applications, however its detection with MRI still has relatively low sensitivity. In this work we present an improved ferritin chimera, genetically engineered to exhibit stronger paramagnetic properties.

Disruption of Foxg1 expression by knock-in of cre recombinase: effects on the development of the mouse telencephalon.

The cre/loxP system is used routinely to manipulate gene expression in the mouse nervous system. In order to delete genes specifically from the telencephalon, the Foxg1-cre line was created previously by replacing the intron-less Foxg1 coding region with cre, resulting in a Foxg1 heterozygous mouse. As the telencephalon of heterozygous Foxg1 mice was reported to be normal, this genotype often has been used as the control in subsequent analyses. Here we describe substantial disruption of forebrain development of heterozygous mice in the Foxg1-cre line, maintained on the C57BL/6J background. High resolution magnetic resonance microscopy reveals a significant reduction in the volume of the neocortex, hippocampus and striatum. The alteration in the neocortex results, in part, from a decrease in its tangential dimension, although gross patterning of the cortical sheet appears normal. This decrease is observed in three different Foxg1 heterozygous mouse lines, independent of the method of achieving deletion of the Foxg1 gene. Although Foxg1 is not expressed in the diencephalon, three-dimensional magnetic resonance microscopy revealed that thalamic volume in the adult is reduced. In contrast, at postnatal day 4, thalamic volume is normal, suggesting that interactions between cortex and dorsal thalamus postnatally produce the final adult thalamic phenotype. In the Foxg1-cre line maintained on the C57BL/6J background, the radial domain of the cerebral cortex also is disrupted substantially, particularly in supragranular layers. However, neither Foxg1 heterozygous mice of the Foxg1-tet (tetracycline transactivator) line, nor those of the Foxg1-lacZ and Foxg1-cre lines maintained on a mixed background, displayed a reduced cortical thickness. Thus Cre recombinase contributes to the radial phenotype, although only in the context of the congenic C57BL/6J background. These observations highlight an important role for Foxg1 in cortical development, reveal noteworthy complexity in the invocation of specific mechanisms underlying phenotypes expressed following genetic manipulations and stress the importance of including appropriate controls of all genotypes.

Comparative chromosomal studies of E. caballus (ECA) and E. przewalskii (EPR) in a female F1 hybrid.

Previous research revealed that the karyotypes of Equus przewalskii (2n = 66) and Equus caballus (2n = 64) differ by one pair of metacentric chromosomes, present in ECA but not in EPR, and two pairs of acrocentric chromosomes found only in the EPR karyotype. The formation of a trivalent during meiosis in a male F1 hybrid and the homologies in G-banding patterns suggest that ECA 5 corresponds to two acrocentric EPR chromosomes resulting from a Robertsonian fusion or fission event. Chromosomal investigations of a female interspecies F1 hybrid including banded karyograms and fluorescence in situ hybridization (FISH) studies focusing on the p and q arm of ECA 5 were conducted. Q- and G-banding patterns of E. caballus, E. przewalskii and the hybrid revealed interspecies homology between all chromosome pairs except for ECA 5, EPR 23 and EPR 24, which were unique for that particular species. Furthermore, they indicated homology between ECA 5p and EPR 23 as well as between ECA 5q and EPR 24. FISH revealed hybridization of the BACs laminin beta 3 (LAM B3) and laminin gamma 2 (LAM C2) to ECA 5p and EPR 23. However, nuclear factor I (NFIA) and immunoglobulin lambda (IGL@), primarily assigned to ECA 5q, mapped to ECA 7 and EPR 6 respectively. Thus the karyotypes of E. caballus and E. przewalskii differ solely by one Robertsonian translocation (ECA 5 =EPR 23 + EPR 24).

Bovine spinal muscular atrophy: AFG3L2 is not a positional candidate gene.

Bovine spinal muscular atrophy (BSMA) is a neurodegenerative disorder, which is widespread in Brown Swiss cattle. Main symptoms of the disease are muscular atrophy and recumbency. Affected calves die within few days or weeks. BSMA seems to be inherited as a recessive trait and the disease allele appears to have a common origin. In this study, a pedigree with 30 affected BSMA calves was used to genetically localize the BSMA locus. Linkage analysis was performed between microsatellite markers of seven chromosomes, where the homologous genes of human neurodegenerative disorders are located according to comparative mapping data, and the disease genotype. BSMA was mapped to chromosome 24 confirming the recently published localization (Medugorac et al. 2003). The candidate gene AFG3L2 was physically mapped to chromosome 24q24 using fluorescence in situ hybridization. Due to their different localizations AFG3L2 is not a positional candidate for BSMA. An informative marker localized on the telomeric side of the BSMA locus would be beneficial for marker-assisted selection as well as searching for the causative gene. However, finding a marker distal to BSMA locus is difficult because of its position at the end of the chromosome.

Cytogenetic comparison of saola (Pseudoryx nghetinhensis) and cattle (Bos taurus) using G- and Q-banding and FISH.

In order to cytogenetically describe the new bovid species saola (Pseudoryx nghetinhensis), comparative G- and Q-banding of saola and cattle (Bos taurus) chromosomes as well as FISH-mapping of 32 type-I markers (29 Texas markers and three additional markers) on saola chromosomes were performed. Saola was shown to have a diploid number of 2n = 50 chromosomes possessing five biarmed autosomal pairs and an acrocentric X chromosome. Homology of saola and cattle chromosomes was indicated by banding patterns and by marker hybridization suggesting that all five biarmed pairs in saola originate from centric fusions involving ten cattle autosomes. However, small intrachromosomal rearrangements cannot be excluded. In this study the first preliminary homology map of these two species is presented.

Development of a novel self-medicating applicator for control of internal and external parasites of wild and domestic animals.

Four trials, three in the United States and one in South Africa, were conducted to evaluate the potential value of a novel self-medicating applicator in the passive control of gastrointestinal nematodes in cattle and deer, and of files and ticks on cattle using oil-based treatments. The results of the trials demonstrated that this applicator is an effective and practical device for the passive treatment of both deer and cattle for trichostrongyle infections using the endectocide, moxidectin (Cydectin. Fort Dodge Animal Health, USA), of cattle for hom fly (Haemotobia irritans) infestations using the insecticide, cyfluthrin (CyLence, Bayer AG, Germany) and of cattle for tick infestations (in particular Amblyomma hebraeum and Rhipicephalus appendiculatus) using the acaricides deltamethrin and amitraz (Delete All, Intervet, South Africa).

Receptor-mediated endocytosis of iron-oxide particles provides efficient labeling of dendritic cells for in vivo MR imaging.

Dendritic cells (DCs) function as antigen presenting cells in vivo and play a fundamental role in numerous diseases. New methods are described for high-efficiency intracellular labeling of DCs with superparamagnetic iron-oxide (SPIO) utilizing a receptor-mediated endocytosis (RME) mechanism. Bone marrow-derived DCs or a fetal skin-derived DC line were incubated with SPIO conjugated to anti-CD11c monoclonal antibody (mAb) under conditions favoring RME. These cells exhibited approximately a 50-fold increase in uptake relative to DCs incubated with SPIO without the mAb. Flow cytometry studies assaying cell surface markers showed a down-modulation of CD11c, but no other changes in phenotype. Immunological function of the DCs was unmodified by the labeling, as determined by cytokine secretion assays. The RME mechanism was confirmed using electron microscopy, endocytosis inhibition assays, and incubation experiments with SPIO conjugated to mAbs against accessory molecules that are not expressed on DCs. Labeled DCs were injected into murine quadriceps and monitored in vivo for several days using MR microimaging at 11.7 T. DCs were observed to remain within the muscle for >24 hr. The use of RME is an efficient way to label immune cells for in vivo MRI and can be applied to a wide variety of cell types.

Peripheral somatosensory fMRI in mouse at 11.7 T.

The feasibility of performing extremely-high resolution somatosensory fMRI in anesthetized mice using BOLD contrast at 11.7 T was investigated. A somatosensory stimulus was applied to the hindlimb of an alpha-chlorolose anesthetized mouse resulting in robust (p < 4 x 10(-3)) BOLD changes in somatosensory cortex and large veins. Percentage modulation of the MR signal in cortex exceeded 7%. Experiments that artificially modulated the inspired oxygen tension were also conducted; the results revealed large, heterogeneous, BOLD contrast changes in the mouse brain. In addition, T(1), T(2), and T(2)* values in gray matter at 11.7 T were evaluated. Discussion of the sensitivity limitations of BOLD fMRI in the tiny mouse central nervous system is presented. These methods show promise for the assessment of neurological function in mouse models of CNS injury and disease.

In vivo observation of cavitation and embolism repair using magnetic resonance imaging.

Magnetic resonance imaging (MRI) was used to noninvasively monitor the status of individual xylem vessels in the stem of an intact, transpiring grape (Vitis vinifera) plant over a period of approximately 40 h. Proton density-weighted MRI was used to visualize the distribution of mobile water in the stem and individual xylem vessels were scored as either water or gas filled (i.e. embolized). The number of water-filled vessels decreased during the first 24 h of the experiment, indicating that approximately 10 vessels had cavitated during this time. Leaf water potentials decreased from -1.25 to -2.1 MPa during the same period. Watering increased leaf water potentials to -0.25 MPa and prevented any further cavitation. Refilling of xylem vessels occurred as soon as the lights were switched off, with the majority of vessels becoming refilled with water during the first 2 to 3 h in darkness. These measurements demonstrate that MRI can be used to monitor the functional status of individual xylem vessels, providing the first method to study the process of cavitation and embolism repair in intact plants.

In vivo visualization of gene expression using magnetic resonance imaging.

High-resolution in vivo imaging of gene expression is not possible in opaque animals by existing techniques. Here we present a new approach for obtaining such images by magnetic resonance imaging (MRI) using an MRI contrast agent that can indicate reporter gene expression in living animals. We have prepared MRI contrast agents in which the access of water to the first coordination sphere of a chelated paramagnetic ion is blocked with a substrate that can be removed by enzymatic cleavage. Following cleavage, the paramagnetic ion can interact directly with water protons to increase the MR signal. Here, we report an agent where galactopyranose is the blocking group. This group renders the MRI contrast agent sensitive to expression of the commonly used marker gene, beta-galactosidase. To cellular resolution, regions of higher intensity in the MR image correlate with regions expressing marker enzyme. These results offer the promise of in vivo mapping of gene expression in transgenic animals and validate a general approach for constructing a family of MRI contrast agents that respond to biological activity.

Control of Boophilus annulatus (Acari: Ixodidae) on cattle using injectable microspheres containing ivermectin.

The efficacy of an injectable microsphere formulation of ivermectin for control of the cattle tick, Boophilus annulatus (Say), was tested on 2 groups of 6 Hereford heifers held on separate 7-ha, tick-infested, buffel grass pastures. Cattle in one pasture were injected subcutaneously in the neck with a controlled-release microsphere formulation of ivermectin at the rate of 2.4 mg AI/kg body weight; the other group was injected with carrier only. Beginning 4 wk after injection and continuing throughout the remainder of the test (16 wk), no engorged ticks (> or = 5.5 mm) were found on any of the treated cattle, whereas large numbers of engorged ticks were found on the untreated controls. During this period, a few ticks were recovered from untreated sentinel animals placed in the treatment pasture during 7-8 wk after treatment, but none were recovered from animals exposed from 11-12 wk or 14-15 wk. Large numbers of B. annulatus ticks were found on untreated sentinel cattle placed in the control pasture during these same periods. Although the cattle, pastures, and tick habitat were approximately equal, the treated cattle gained an average of 77 kg compared with an average of 42 kg for the control group. This technology offers a possible alternative to the current official program of dipping and vacating pastures for eradication of Boophilus sp. infestations from the quarantine zone in southern Texas. Larger scale testing is needed to determine the potential of the injectable microsphere formulation and to optimize its use in eradication or control strategies.

Looking deeper into vertebrate development.

Magnetic resonance imaging (MRI) is a non-invasive imaging method that provides three-dimensional (3-D) images of the internal structure of opaque objects, such as humans and mice. In optimal situations, spatial resolution can approach the micron level. Arbitrarily oriented single-slice images can be obtained in seconds, with full 3-D volume images taking tens of minutes to collect. The exquisite sensitivity of MRI to the local physical and chemical environment provides a wide range of mechanisms giving rise to intrinsic contrast in the MR experiment, thus providing images with dramatic differences between different tissue types (e.g. white vs grey matter, myelinated vs unmyelinated fibres, and brain parenchyma vs ventricles). The recent advent of physiologically sensitive MRI contrast agents opens up a wealth of new avenues of study, even including the in vivo imaging of gene expression.

Towards a microMRI atlas of mouse development.

This study investigates the potential of microscopic Magnetic Resonance Imaging to obtain information for 3D digital atlases of mouse development using fixed samples. Fixed samples allow direct comparison with already published atlases and provide a testing ground for future in vivo efforts. 3D MR images of mouse embryos (dpc 6.5-16) illustrate that the necessary contrast and level of detail is available with this technique. Diffusion weighted imaging, diffusion tensor imaging, and multi-valued data sets are presented as examples of uniquely MR methods of obtaining anatomical information. MRI is performed non-invasively on the intact sample, leaving open the possibility of other manipulations (e.g. classical histology, immunohistochemistry, in situ hybridization, and in vitro growth for unfixed samples) after conducting the MRI experiment.

A model for MRI contrast enhancement using T1 agents.

Contrast in MRI relies on differences in the local environment of water and is often enhanced by using contrast agents. We present a simple model for evaluating the minimal contrast agent concentration required to produce "satisfactory" contrast enhancement in magnetic resonance images. Previous strategies have been based largely on empirical results for specific systems. The present tissue contrast model (TCM) can be applied to "conventional," targeted, or biochemically responsive agents. The model results are formulated so that only a small number of parameters are required to analyze a given scenario. The TCM is a particularly useful tool in the development of new classes of magnetic resonance contrast media. These agents will have the ability to target specific cells or tissue, and perhaps be able to report on their physiological status. As an example of the applicability of the TCM, we test it against in vivo magnetic resonance microscopy results in frog embryos that have focal cell populations labeled with contrast agent by using calibrated single-cell microinjection techniques.

MR microscopy of transgenic mice that spontaneously acquire experimental allergic encephalomyelitis.

Pathology of fixed spinal cords from transgenic mice with a myelin basic protein (MBP) specific T cell receptor was investigated. These mice spontaneously acquire the demyelinating disease experimental allergic encephalomyelitis (EAE). Several complementary imaging modalities, all on the same tissues, were used to visualize lesions; these included high-field (11.7-T) microscopic diffusion tensor imaging (DTI), T2*-weighted imaging, and optical microscopy on histological sections. Lesions were predominantly in white matter around meninges and vasculature and appeared hyperintense in anatomical images. DTIs showed reduced diffusion anisotropy in the same hyperintense regions, consistent with inflammation and edema. Histology in the same tissues exhibited the characteristic pathology of EAE. Two techniques for visualizing the effective diffusion tensor fields are presented, which display direction, organization, and integrity of neuronal fibers. It is shown that DTI offers intriguing possibilities for visualizing axonal organization and lesions within white matter.

Therapeutic and persistent efficacy of fipronil against Boophilus microplus (Acari: Ixodidae) on cattle.

Cattle infested with all parasitic life-stages of Boophilus microplus (Canestrini) were treated once with a pour-on formulation of fipronil at 0.25, 0.5, or 1.0% active ingredient (AI) to determine the therapeutic efficacy. Additionally, cattle were infested at weekly intervals for 13 consecutive weeks after treatment to determine the persistent effectiveness of the three concentrations. Results of therapeutic efficacy of fipronil showed that with each increase in concentration there was a corresponding and often significant (P < 0.05) decrease in the numbers, fecundity and fertility of engorged females. The overall control of the Index of Reproduction (IR) achieved at 0.25 and 0.5% AI was 86.2 and 94.3%, respectively, while the control of the IR at the 1.0% AI concentration was 99.7%. The persistent efficacy of fipronil, like the therapeutic efficacy, was also dose related. The 0.25% AI treatment failed to provide complete protection (100%) against larval reinfestation, although control of the IR remained high (> 95%) for the first 6 weeks after treatment. In contrast, at 0.5% AI the control of the IR remained at 100% for the first 4 weeks after treatment, and was > 90% for an additional 2 weeks (6 weeks posttreatment). The 1.0% AI concentration afforded the longest residual activity, providing virtually 100% protection against larval reinfestation for 8 weeks after treatment was applied. Based on the results, neither the 0.25 or 0.5% AI concentration completely eliminated the immature stages of the tick (nymphs and larvae) on the host at the time of treatment (therapeutic efficacy), and the residual activity (persistent efficacy) did not prevent reinfestation by larvae for long periods after treatment. While the 1.0% AI concentration provided both therapeutic and residual efficacy that would be suitable for use in the eradication program, caution is still warranted because a few engorged females detached for up to 25 days after treatment, although none reproduced after the 6th day.

Efficacy of amitraz (Taktic 12.5% EC) as a dip for the control of Boophilus microplus (Canestrini) (Acari: Ixodidae) on cattle.

Four groups of cattle infested with Boophilus microplus (Canestrini) were each dipped in a different concentration of amitraz diluted from a 12.5% EC formulation to determine the efficacy and performance of the product in an 11,400 l dipping vat. Except for the period when heifers were dipped, animals were restrained in stanchions placed individually inside 3.3 x 3.3 m2 stalls within an open-sided barn. The amitraz in the vat was stabilized with hydrated lime to maintain a pH of ca. 12. Analyses of vat samples showed that concentrations of amitraz in the vat were 7.6 to 13% lower than the targeted concentrations of 0.010, 0.015, 0.020, and 0.025% active ingredient (AI) for dilutions prepared according to instructions on the manufacturer's label. The large quantity of hydrated lime added to the vat (10 kg/1000 l) interfered with the HPLC analysis of vat samples. Therapeutic efficacy of each of the four observed concentrations (0.0088, 0.0131, 0.0174, and 0.0231% AI) of amitraz was excellent (> 99% control). However, the rapid detachment of all ticks from an animal within a few hours after treatment with amitraz, that has been frequently observed, was not pronounced in the present study. Only 47% of the B. microplus detached in the first 4 h post-treatment, and 84% detached within the first 24 h. All of the treatments, except the lowest concentration, provided protection of cattle against re-infestation by B. microplus larvae for 14 days post-treatment. Possibly as a result of the formation of a compact layer of lime and amitraz on the bottom after the vat was undisturbed for six weeks, intense agitation was required to re-suspend the active ingredient.