Do improvements in clinical practice guidelines alter pregnancy outcomes in asthmatic women? A single-center retrospective cohort study.

OBJECTIVE: Asthma occurs in ∼17% of Australian pregnancies and is associated with adverse perinatal outcomes, which worsen with poor asthma control. Consequently, the South Australian 'Asthma in Pregnancy' perinatal guidelines were revised in 2012 to address management according to severity. This study investigated if these revised guidelines reduced the impact of maternal asthma on risks of adverse perinatal outcomes before (Epoch 1, 2006-2011) and after the revision (Epoch 2, 2013-2018). METHODS: Routinely collected perinatal and neonatal datasets from the Women's and Children's Hospital (Adelaide, Australia) were linked. Maternal asthma (prevalence:7.5%) was defined as asthma medication use or symptoms described to midwives. In imputation (n = 59131) and complete case datasets (n = 49594), analyses were conducted by inverse proportional weighting and multivariate logistic regression, accounting for confounders. RESULTS: Overall, maternal asthma was associated with increased risks of any antenatal corticosteroid treatment for threatened preterm birth (aOR 1.319, 95% CI 1.078-1.614), any Cesarean section (aOR 1.196, 95% CI 1.059-1.351), Cesarean section without labor (aOR 1.241, 95% CI 1.067-1.444), intrauterine growth restriction (IUGR, aOR 1.285, 95% CI 1.026-1.61), and small for gestational age (aOR 1.324, 95% CI 1.136-1.542). After guideline revision, asthma-associated risks of any Cesarean section (p < 0.001), any antenatal corticosteroids (p = 0.041), and small for gestational age (p = 0.050), but not IUGR and Cesarean section without labor, were reduced. CONCLUSIONS: Clinical practice guidelines based on the latest evidence do not guarantee clinical efficacy. Since adverse perinatal outcomes did not all improve, this work highlights the need to evaluate the ongoing impact of guidelines on clinical outcomes.

Salivary antibodies are detected with a commercial anti-SARS-CoV-2 assay only after two doses of vaccine using serum thresholds.

Salivary matrix is an appealing specimen type for SARS-CoV-2 serology because of ease of collection and potential for concurrent nucleic acid testing. We address the feasibility of salivary matrix to detect anti-SARS-CoV-2 antibodies using two commercially available anti-SARS-CoV-2 Total antibody assays including analytical validations. Matched serum and saliva samples were collected from 10 convalescent COVID-19 patients and tested using a quantitative anti-Spike Total antibody assay and a qualitative anti-Nucleocapsid Total antibody assay from Roche Diagnostics. Both assays were 100% sensitive for COVID-19 history in serum. However, saliva samples were below serum positivity thresholds. We then collected longitudinal salivary samples from a volunteer cohort receiving the Pfizer-BioNTech COVID-19 BNT162b2 vaccine. Saliva was negative for anti-SARS-CoV-2 antibodies at 5 time points after a single dose of vaccine including day 56 when mean (min-max) serum levels of anti-Spike Total antibody were 79.0 U/mL (46.6-110.1) (N = 8). After a second vaccine dose serum-matched samples were beyond the analytical measuring range of the assay (>2500 U/mL), and detection of salivary anti-Spike Total antibody was achieved in all volunteers (12.2 U/mL [2.0-32.7]) (N = 11) 30 days after the second dose. Mean anti-Spike Total antibody levels in serum (1558 U/mL (434->2500)) and saliva (2.6 U/mL (<0.4-11.4)) declined 216-233 days after the first dose of vaccine (P < 0.05); and saliva was 75% sensitive for two doses of vaccination at this latter time point (N = 25). These data suggest commercial assays are capable of detecting vaccine status after two doses of BNT162b2 vaccine up to 6 months and could inform COVID-19 surveillance.

Estrogen receptor alpha mediates mandibular condylar cartilage growth in male mice.

OBJECTIVES: In the appendicular skeleton, estrogen via ERα signalling has been shown to mediate endochondral growth plate fusion in both males and females. However, the role of ERα in mediating growth of the mandibular condylar cartilage is unknown. Thus, this study focuses on the characterization of the mandibular condylar cartilage phenotype in young and adult male ERαKO mice. SETTING: Columbia University Medical Center. MATERIAL AND METHODS: WT and ERαKO C57BL/6 male mice were sacrificed at 49 days or 9 months for phenotypic analysis. Changes to MCC thickness, cell number and cell density were measured using histomorphometric methods. Cartilage-specific gene expression and OARSI scores were investigated for 49-day and 9-month-old male ERαKO and WT mice. RESULTS: In young mice, a significant increase in the number of mandibular condylar cartilage cells and a significant decrease in the expression of Col10, Runx2 and DMP1 were observed in the male ERαKO mice compared to WT. In 9-month-old mice, we found a similar increase in the number of cells but no change in osteoarthritic histological scoring in ERαKO mice compared to WT mice. CONCLUSION: In summary, estrogen plays a role in mediating mandibular condylar maturation in young male mice. However, according to this study, it does not play a role in mediating long-term growth or age-related mandibular condylar cartilage degeneration in males.

Novel conformation-selective alpha-synuclein antibodies raised against different in vitro fibril forms show distinct patterns of Lewy pathology in Parkinson's disease.

AIMS: The aim of this study was to test the hypothesis that different conformations of misfolded α-synuclein (α-syn) are present in Parkinson's disease (PD) brain. METHODS: Using two previously characterized conformations of α-syn fibrils, we generated new conformation-selective α-syn monoclonal antibodies (mAbs). We then interrogated multiple brain regions in a well-characterized autopsy cohort of PD patients (n = 49) with these mAbs, Syn7015 and Syn9029. RESULTS: Syn7015 detects Lewy bodies (LBs) and Lewy neurites (LNs) formed by pathological α-syn in all brain regions tested, and is particularly sensitive to LNs and small Lewy dots, inclusions believed to form early in the disease. Further, we observed colocalization between Syn7015 and an early marker of α-syn pathology formation, phospho-Ser129-α-syn, and a lack of extensive colocalization with markers of more mature pathology. In comparison, Syn9029 detects Lewy pathology in all regions examined, but indicates significantly fewer LNs than Syn7015. In addition, colocalization of Syn9029 with later markers of α-syn pathology maturation (ubiquitin and P62) suggests that the pathology detected by Syn9029 is older. Semiquantitative scoring of both LN and LB pathology in nine brain regions further established this trend, with Syn7015 LN scores consistently higher than Syn9029 LN scores. CONCLUSIONS: Our data indicate that different conformations of α-syn pathology are present in PD brain and correspond to different stages of maturity for Lewy pathology. Regional analysis of Syn7015 and Syn9029 immunostaining also provides support for the Braak hypothesis that α-syn pathology advances through the brain.

Comparison of In Vivo and Ex Vivo MRI of the Human Hippocampal Formation in the Same Subjects.

Multiple techniques for quantification of hippocampal subfields from in vivo MRI have been proposed. Linking in vivo MRI to the underlying histology can help validate and improve these techniques. High-resolution ex vivo MRI can provide an intermediate modality to map information between these very different imaging modalities. This article evaluates the ability to match information between in vivo and ex vivo MRI in the same subjects. We perform rigid and deformable registration on 10 pairs of in vivo (3 T, 0.4 × 0.4 × 2.6 mm3) and ex vivo (9.4 T, 0.2 × 0.2 × 0.2 mm3) scans, and describe differences in MRI appearance between these modalities qualitatively and quantitatively. The feasibility of using this dataset to validate in vivo segmentation is evaluated by applying an automatic hippocampal subfield segmentation technique (ASHS) to in vivo scans and comparing SRLM (stratum/radiatum/lacunosum/moleculare) surface to manual tracing on corresponding ex vivo scans (and in 2 cases, histology). Regional increases in thickness are detected in ex vivo scans adjacent to the ventricles and were not related to scanner, resolution differences, or susceptibility artefacts. Satisfactory in vivo/ex vivo registration and subvoxel accuracy of ASHS segmentation of hippocampal SRLM demonstrate the feasibility of using this dataset for validation, and potentially, improvement of in vivo segmentation methods.

Sex differences in the estrogen-dependent regulation of temporomandibular joint remodeling in altered loading.

OBJECTIVE: Temporomandibular joint (TMJ) diseases predominantly afflict women, suggesting a role of estrogen in the disease etiology. Previously, we determined that decreased occlusal loading (DOL) inhibited collagen type II (Col2) expression in the mandibular condylar cartilage (MCC) of female wild-type (WT) mice whereas no change was observed in males. This decrease in chondrogenesis was abolished by estrogen receptor beta (ERß) deficiency in females. Therefore, the goal of this study was to examine the role of estradiol - ERß signaling in mediating DOL effects in male mice to further decipher sex differences. METHODS: Male 21 day-old WT and ERßKO male mice were treated with either placebo or estradiol and exposed to normal or DOL for 4 weeks. Cartilage thickness and cell proliferation, gene expression and immunohistochemistry of chondrogenic markers and estrogen receptor alpha (ERα), and analysis of bone histomorphometry via microCT were completed to ascertain the effect of estradiol on DOL effects to the TMJ. RESULTS: ERßKO male mice lack a MCC phenotype. In both genotypes, estradiol treatment increased Col2 gene expression and trabecular thickness. DOL in combination with estradiol treatment caused a significant increase in Col2 gene expression in both genotypes. CONCLUSIONS: The sex differences in DOL-induced inhibition of Col2 expression do not appear to be mediated by differences in estradiol levels between male and female mice. Greater understanding on the role of estrogen and altered loading are critical in order to decipher the sex dimorphism of TMJ disorders.

Progression of alpha-synuclein pathology in multiple system atrophy of the cerebellar type.

AIMS: The aim of this study was to identify early foci of α-synuclein (α-syn pathology) accumulation, subsequent progression and neurodegeneration in multiple system atrophy of the cerebellar type (MSA-C). METHODS: We analysed 70-µm-thick sections of 10 cases with MSA-C and 24 normal controls. RESULTS: MSA-C cases with the lowest burden of pathology showed α-syn glial cytoplasmic inclusions (GCIs) in the cerebellum as well as in medullary and pontine cerebellar projections. Cerebellar pathology was highly selective and severely involved subcortical white matter, whereas deep white matter and granular layer were only mildly affected and the molecular layer was spared. Loss of Purkinje cells increased with disease duration and was associated with neuronal and axonal abnormalities. Neocortex, basal ganglia and spinal cord became consecutively involved with the increasing burden of α-syn pathology, followed by hippocampus, amygdala, and, finally, the visual cortex. GCIs were associated with myelinated axons, and the severity of GCIs correlated with demyelination. CONCLUSIONS: Our findings indicate that cerebellar subcortical white matter and cerebellar brainstem projections are likely the earliest foci of α-syn pathology in MSA-C, followed by involvement of more widespread regions of the central nervous system and neurodegeneration with disease progression.

The use of the Rx spin label in orientation measurement on proteins, by EPR.

The bipedal spin label Rx is more restricted in its conformation and dynamics than its monopodal counterpart R1. To systematically investigate the utility of the Rx label, we have attempted to comprehensively survey the attachment of Rx to protein secondary structures. We have examined the formation, structure and dynamics of the spin label in relation to the underlying protein in order to determine feasibility and optimum conditions for distance and orientation measurement by pulsed EPR. The labeled proteins have been studied using molecular dynamics, CW EPR, pulsed EPR distance measurement at X-band and orientation measurement at W-band. The utility of different modes and positions of attachment have been compared and contrasted.

Correction: In situ crosslinking of electrospun gelatin for improved fiber morphology retention and tunable degradation.

Correction for 'In situ crosslinking of electrospun gelatin for improved fiber morphology retention and tunable degradation' by A. P. Kishan et al., J. Mater. Chem. B, 2015, DOI: .

In situ crosslinking of electrospun gelatin for improved fiber morphology retention and tunable degradation.

Electrospinning is a popular technique to fabricate tissue engineering scaffolds due to the exceptional tunability of the fiber morphology, which can be used to control the scaffold mechanical properties, degradation rate, and cell behavior. Recent work has focused on electrospinning natural polymers such as gelatin to improve the regeneration potential of these grafts. Gelatin scaffolds must be crosslinked to avoid rapid dissolution upon implantation with current crosslinking strategies requiring additional post-processing steps. Despite the strong dependence of scaffold properties on fiber morphology, there has been minimal emphasis on retaining the original fiber morphology of electrospun gelatin scaffolds after implantation. This work describes a method for in situ crosslinking of gelatin to produce electrospun fibers with improved fiber morphology retention after implantation. A double barrel syringe with an attached mixing head and a diisocyanate crosslinker were utilized to generate electrospun scaffolds that crosslink during the electrospinning process. These in situ crosslinked fiber meshes retained morphology after 1 week incubation in water at 37 °C; whereas, uncrosslinked meshes lost the fibrous morphology within 24 hours. Degree of crosslinking was quantified and relationships between the crosslinker ratio and enzymatic degradation rate were evaluated. The degradation rate decreased with increased crosslinker ratio, resulting in a highly tunable system. Additionally, tensile testing under simulated physiological conditions indicated that increased crosslinker ratios resulted in increases in initial modulus and tensile strength. Overall, this in situ crosslinking technique provides a method to crosslink gelatin during electrospinning and can be used to tune the degradation rate of resulting scaffolds while enabling improved fiber morphology retention after implantation.

Elevated levels of FOXA1 facilitate androgen receptor chromatin binding resulting in a CRPC-like phenotype.

Castration-resistant prostate cancer (CRPC) continues to pose a significant clinical challenge with new generation second-line hormonal therapies affording limited improvement in disease outcome. As the androgen receptor (AR) remains a critical driver in CRPC, understanding the determinants of its transcriptional activity is important for developing new AR-targeted therapies. FOXA1 is a key component of the AR transcriptional complex yet its role in prostate cancer progression and the relationship between AR and FOXA1 are not completely resolved. It is well established that FOXA1 levels are elevated in advanced prostate cancer and metastases. We mimicked these conditions by overexpressing FOXA1 in the androgen-responsive LNCaP prostate cancer cell line and observed a significant increase in AR genomic binding at novel regions that possess increased chromatin accessibility. High levels of FOXA1 resulted in increased proliferation at both sub-optimal and high 5α-dihydrotestosterone (DHT) concentrations. Immunohistochemical staining for FOXA1 in a clinical prostate cancer cohort revealed that high FOXA1 expression is associated with shorter time to biochemical recurrence after radical prostatectomy (hazard ratio (HR) 5.0, 95% confidence interval (CI) 1.2-21.1, P=0.028), positive surgical margins and higher stage disease at diagnosis. The gene expression program that results from FOXA1 overexpression is enriched for PTEN, Wnt and other pathways typically represented in CRPC gene signatures. Together, these results suggest that in an androgen-depleted state, elevated levels of FOXA1 enhance AR binding at genomic regions not normally occupied by AR, which in turn facilitates prostate cancer cell growth.

Minireview: The androgen receptor in breast tissues: growth inhibitor, tumor suppressor, oncogene?

Androgen receptor (AR) signaling exerts an antiestrogenic, growth-inhibitory influence in normal breast tissue, and this role may be sustained in estrogen receptor α (ERα)-positive luminal breast cancers. Conversely, AR signaling may promote growth of a subset of ERα-negative, AR-positive breast cancers with a molecular apocrine phenotype. Understanding the molecular mechanisms whereby androgens can elicit distinct gene expression programs and opposing proliferative responses in these two breast cancer phenotypes is critical to the development of new therapeutic strategies to target the AR in breast cancer.

Epidemiology of vancomycin-resistant enterococci in children with acute lymphoblastic leukemia at two referral centers in Tehran, Iran: a descriptive study.

BACKGROUND: Risk factors for colonization with vancomycin-resistant enterococci (VRE) vary by population and locale. The objective of this study was to determine the prevalence of and risk factors for VRE colonization in children with acute lymphoblastic leukemia (ALL) in Tehran. METHODS: Stools were collected from children with ALL at the Ali Asghar Children's Hospital and the Mahak Pediatric Oncology Center between March 2007 and October 2008. Demographic features and potential risk factors for VRE colonization, including duration of ALL, presence of severe neutropenia in the preceding month, receipt of antibiotics in the preceding 3 months, concurrent medical problems, days of hospitalization, and the need for intensive care since the time of diagnosis of ALL, were recorded. RESULTS: VRE was identified from stools in 33 of 130 children with ALL (25%). No clear risk factors were identified for VRE colonization in the current study, but there was a trend towards an increased prevalence in children admitted to the intensive care unit since their ALL diagnosis (p=0.07). The VanA genotype was found in 28 of the 33 stools (85%), with all other enterococci being VanB. CONCLUSIONS: The prevalence of VRE colonization in children with ALL in Tehran is high. Modifiable risk factors have not been identified. The implementation of routine surveillance for colonization and an increased emphasis on adherence to standard infection control precautions may prevent spread.

Variants in several genomic regions associated with asperger disorder.

Asperger disorder (ASP) is one of the autism spectrum disorders (ASD) and is differentiated from autism largely on the absence of clinically significant cognitive and language delays. Analysis of a homogenous subset of families with ASP may help to address the corresponding effect of genetic heterogeneity on identifying ASD genetic risk factors. To examine the hypothesis that common variation is important in ASD, we performed a genome-wide association study (GWAS) in 124 ASP families in a discovery data set and 110 ASP families in a validation data set. We prioritized the top 100 association results from both cohorts by employing a ranking strategy. Novel regions on 5q21.1 (P = 9.7 × 10(-7) ) and 15q22.1-q22.2 (P = 7.3 × 10(-6) ) were our most significant findings in the combined data set. Three chromosomal regions showing association, 3p14.2 (P = 3.6 × 10(-6) ), 3q25-26 (P = 6.0 × 10(-5) ) and 3p23 (P = 3.3 × 10(-4) ) overlapped linkage regions reported in Finnish ASP families, and eight association regions overlapped ASD linkage areas. Our findings suggest that ASP shares both ASD-related genetic risk factors, as well as has genetic risk factors unique to the ASP phenotype.

Vitamin D status is not associated with the risk of hospitalization for acute bronchiolitis in early childhood.

The association between vitamin D status and susceptibility to acute lower respiratory tract infection (ALRI) was studied in young Canadian children. Serum 25-hydroxyvitamin D (25(OH)D) concentrations were measured in patients aged 1-25 months admitted to hospital with uncomplicated ALRI (primarily viral bronchiolitis) as well as in healthy, similarly aged patients without a history of hospitalization for ALRI (controls). Serum 25(OH)D concentrations were similar among cases and controls (77.0 versus 77.2 nmol l(-1); P=0.960), and there was no case-control difference in the prevalence of vitamin D insufficiency using two thresholds (<40 nmol l(-1): 4.7 versus 1.5%, P=0.365; <80 nmol l(-1): 51.6 versus 56.9%, P=0.598). Vitamin D status was not associated with the risk of hospitalization for ALRI in this population.

Respiratory failure associated with human metapneumovirus infection in an infant posthepatic transplant.

This is the first reported case of respiratory failure associated with human metapneumovirus (hMPV) infection in a liver transplant recipient or in a pediatric solid transplant recipient. A 9-month-old female developed respiratory distress 8 days following a liver transplant. hMPV was detected and she required intubation followed by extracorporeal membrane oxygenation for 26 days. Immunosuppressive medications were stopped during the acute infection except for methylprednisolone as treatment for acute respiratory distress. Serial Doppler ultrasounds were used to monitor for hepatic vessel thromboses and serum liver function tests to assess for hepatic dysfunction and there was no evidence of allograft rejection. The patient recovered from the nosocomial hMPV infection with satisfactory pulmonary function and possible mild developmental delay.

Ontogeny and chain-length specificity of gastrointestinal lipases affect medium-chain triacylglycerol utilization by newborn pigs.

Ontogeny and fatty acid chain-length specificity of gastrointestinal lipases in neonatal piglets were examined to explore the basis for variations in postnatal use of medium-chain triacylglycerols (MCT). Twenty-four newborn pigs were studied at 4 ages: 0, 6, 18, and 48 h postpartum (n = 6 pigs/age). Piglets were gastrically intubated and given 3.0 mmol/kg of BW(0.75) each of emulsified tri-C6:0 and tri-C8:0. One hour after intubation, the plasma concentration of C6:0 was 7.5-fold greater than that of C8:0 (P < 0.001), with total plasma medium-chain fatty acid concentrations 3.7-fold greater at 48 h than at 6 h of age (P < 0.05). Pancreatic, gastric, and lingual tissues were analyzed for lipase activity using an equimolar mixture of tri-C6:0 and tri-C8:0 as substrate. Pancreatic lipase activity averaged 7.0 +/- 0.8 micromol of fatty acid released/min per mg of protein for the medium-chain fatty acid substrates. Hexanoate (C6:0) release was greater at 0 h than at 6, 18, or 48 h (P < 0.05); however, age did not affect C8:0 release (P > 0.05). The lowest lipase activity was observed at 18 h for both tri-C6:0 and tri-C8:0. Chain-length specificity of pancreatic lipase was measured with tri-C4:0, tri-C6:0, tri-C8:0, and tri-C10:0 as combined or separate substrates. As separate substrates, the lipase activity decreased progressively as chain length increased from tri-C4:0 to tri-C10:0. As combined substrates, tri-C6:0 was hydrolyzed fastest (P < 0.05), followed by C4:0, C8:0, and C10:0. Gastric and lingual lipase activities averaged 2.7 nmol/min per mg of protein for the medium-chain fatty acid substrates, with hydrolysis of C6:0 being 7-fold greater than that of C8:0. In conclusion, pancreatic lipase dominates the preduodenal lipases in the neonatal pig, and greater activity of the gastrointestinal lipases toward tri-C6:0 underlies its increased rate of use.

Identification of a mutation associated with factor XI deficiency in Holstein cattle.

An autosomal recessive deficiency of blood coagulation factor XI (FXI) has been described in Holstein cattle. Current testing methods are unsuitable for accurately identifying carriers (heterozygotes) of the disease. To identify the molecular basis of this deficiency, a polymerase chain reaction (PCR)-based strategy was implemented to clone and sequence the bovine FXI gene (F11) from animals of different genotypes. Approximately 14 kb of genomic DNA sequence and 1.8 kb of cDNA sequence, corresponding to exon 3 through the 3'-UTR, of the bovine gene were obtained. Comparison of sequences derived from homozygous normal and deficient individuals revealed that FXI deficiency in Holsteins is associated with the insertion of a 76 bp segment [AT(A)(28)TAAAG(A)(26)GGAAATAATAATTCA] within exon 12. This insertion introduces a stop codon that results in a mature FXI protein lacking the functional protease domain encoded by exons 13, 14 and 15. Based on these data, a DNA-based diagnostic test has been developed for accurate genotyping. Using this method, the frequency of the mutated allele has been determined to be 1.2% in a contemporary population of the USA Holstein sires.

Case report and literature review of late-onset group B streptococcal disease manifesting as necrotizing fasciitis in preterm infants: is this a new syndrome?

We describe a 5-month-old preterm female infant who presented with necrotizing fasciitis involving the face and neck caused by group B streptococcus (GBS). Because of the extent and anatomic location of the necrosis, surgical debridement was delayed for 16 days, but the infant survived. Review of the literature demonstrated that 3 of the 10 previously reported cases of necrotizing fasciitis caused by GBS involved preterm infants and that 2 of these cases also involved the head and neck.