Sex differences in a murine model of asthma are time and tissue compartment dependent.

CONCLUSION: Sexual dimorphism in lung inflammation is both time and tissue compartment dependent. Spatiotemporal variability in sex differences in a murine model of asthma must be accounted for when planning experiments to model the sex bias in allergic inflammation.

Database Tracking in Gender-Affirming Surgery: Are Patients Falling Through the Cracks?

BACKGROUND: This study sought to examine risk factors for venous thromboembolism in transfeminine vaginoplasty. Secondarily, the authors outline reasons why patients are not adequately classified for research purposes despite using relevant queried codes. METHODS: Transgender patients undergoing vaginoplasty were identified with diagnostic and procedure-specific codes using a national surgical database from 2010 through 2019. RESULTS: There were 457 transgender vaginoplasties performed, with 24 wound dehiscences, 17 unplanned reoperations, and 12 surgical site infections. With zero cases of venous thromboembolism, risk factor analysis was deferred. CONCLUSIONS: Heterogeneity in coding practices for gender-affirming surgery led to an uncharacteristically small cohort of transfeminine vaginoplasty patients captured in the database. Current diagnostic and procedure-specific codes are nonspecific and unbundled, hindering accurate assessment of the incidence of standard surgical complications.

Risk Factors for Acute Postoperative Complications Following Operative Management of Le Fort Fractures-A NSQIP Study.

INTRODUCTION: Le Fort fractures comprise a pattern of complex midfacial fractures that arise secondarily to craniofacial trauma. Although management of these fractures has been detailed within the literature, there is a paucity of research examining postoperative outcomes after surgical repair. The primary aim of this study is to assess patient outcomes after operative management of Le Fort fractures, and examine factors influencing the risk for developing postoperative complications, through utilization of the ACS-NSQIP database. METHODS: The American College of Surgeons National Surgical Quality Improvement Program (ACS-NSQIP) database was queried for cases recorded between 2012 and 2019 with International Classification of Disease (ICD)-9 and ICD-10 codes corresponding to Le Fort fractures. Patient demographics, clinical variables, and postoperative variables were recorded. Logistic regression analysis was conducted to identify independent risk factors for postoperative complications. RESULTS: Identification of cases with appropriate ICD codes, and exclusion of those with missing data, yielded 562 patients for analysis. There were no cases of minor complications and 14 cases of severe complications (3 cases of wound dehiscence, 3 cases of transfusion requirement, 1 case of failure to wean from the ventilator for more than 48 h, 1 pulmonary embolism, and 8 cases of reoperation), corresponding to an overall complication rate of 2.49%. Logistic regression analysis revealed steroid use as an independent predictor of severe postoperative complications (OR =13.73, 95% CI: 1.08-128.02, P =0.02). CONCLUSION: The present study is the first to conduct a risk factor analysis of patients with Le Fort fractures using the ACS-NSQIP national database. The overall postoperative complication rate was 2.49%, with 14 cases of complications recorded in 8 years. Although this may suggest that surgical management of Le Fort fractures is generally well-tolerated, it should be noted that this problem is frequently associated with other severe injuries of the head and neck that may influence patient prognosis. Given this, further analysis would benefit from a larger patient cohort and longer postoperative data as the ACS-NSQIP database only records outcomes within 30 days.

More than neutrophils: Lin(+)Ly6G(+)IL-5Rα(+) multipotent myeloid cells (MMCs) are dominant in normal murine bone marrow and retain capacity to differentiate into eosinophils and monocytes.

Bone marrow is a hematopoietic site harboring multiple populations of myeloid cells in different stages of differentiation. Murine bone marrow eosinophils are traditionally identified by Siglec-F(+) staining using flow cytometry, whereas neutrophils are characterized by Ly6G(+) expression. However, using flow cytometry to characterize bone marrow hematopoietic cells in wild-type mice, we found substantial gray areas in identification of these cells. Siglec-F(+) mature eosinophil population constituted only a minority of bone marrow Lin(+)CD45(+) pool (5%). A substantial population of Siglec-F(-) cells was double positive for neutrophil marker Ly6G and eosinophil lineage marker, IL-5Rα. This granulocyte population with mixed neutrophil and eosinophil characteristics is typically attributable to neutrophil pool based on neutral granule staining and expression of Ly6G and myeloid peroxidase. It is distinct from Lineage(-) myeloid progenitors or Siglec-F(+)Ly6G(+) maturing eosinophil precursors, and can be accurately identified by Lineage(+) staining and positive expression of markers IL-5Rα and Ly6G. At 15-50% of all CD45(+) hematopoietic cells in adult mice (percentage varies by sex and age), this is a surprisingly dominant population, which increases with age in both male and female mice. RNA-seq characterization of these cells revealed a complex immune profile and the capacity to secrete constituents of the extracellular matrix. When sorted from bone marrow, these resident cells had neutrophilic phenotype but readily acquired all characteristics of eosinophils when cultured with G-CSF or IL-5, including expression of Siglec-F and granular proteins (Epx, Mbp). Surprisingly, these cells were also able to differentiate into Ly6C(+) monocytes when cultured with M-CSF. Herein described is the discovery of an unexpected hematopoietic flexibility of a dominant population of multipotent myeloid cells, typically categorized as neutrophils, but with the previously unknown plasticity to contribute to mature pools of eosinophils and monocytes.

3D Numerical Modeling of Laser Assisted Tape Winding Process of Composite Pressure Vessels and Pipes-Effect of Winding Angle, Mandrel Curvature and Tape Width.

Advanced thermoplastic composites manufacturing using laser assisted tape placement or winding (LATP/LATW) is a challenging task as monitoring and predicting nip point (bonding) temperature are difficult especially on curved surfaces. A comprehensive numerical analysis of the heat flux and temperature distribution near the nip point is carried out in this paper for helical winding of fiber reinforced thermoplastic tapes on a cylindrically shaped mandrel. An optical ray-tracing technique is coupled with a numerical heat transfer model in the process simulation tool. The developed optical-thermal model predictions were compared with experimental data available in literature to validate its effectiveness. The influences of winding/placement angle, mandrel curvature and tape width on the incident angles, the laser absorbed intensity, and the process temperature distribution are studied extensively using the validated model. Winding/placement angle has a considerable effect on the temperature distribution. Increase in winding angle results in a higher temperature for tape due to more reflections coming from the substrate. On the other hand, substrate temperature decreases as the winding angle increases due to a decrease in the laser incident angles based on the local surface curvature. An increase in mandrel curvature results in higher nip point temperatures for substrate and lower one for tape. Different mandrel sizes for 90 ∘ placement path do not have a strong effect on the substrate process temperature as for other winding angles because of less curvature change of the corresponding irradiated area. Tape width causes local temperature variations at the edges of the tape/substrate. In order to obtain the desired process temeprature during LATW or LATP processes, the laser intensity distribution on the tape and substrate surfaces should be regulated.

Eosinophil accumulation in postnatal lung is specific to the primary septation phase of development.

Type 2 immune cells and eosinophils are transiently present in the lung tissue not only in pathology (allergic disease, parasite expulsion) but also during normal postnatal development. However, the lung developmental processes underlying airway recruitment of eosinophils after birth remain unexplored. We determined that in mice, mature eosinophils are transiently recruited to the lung during postnatal days 3-14, which specifically corresponds to the primary septation/alveolarization phase of lung development. Developmental eosinophils peaked during P10-14 and exhibited Siglec-Fmed/highCD11c-/low phenotypes, similar to allergic asthma models. By interrogating the lung transcriptome and proteome during peak eosinophil recruitment in postnatal development, we identified markers that functionally capture the establishment of the mesenchymal-epithelial interface (Nes, Smo, Wnt5a, Nog) and the deposition of the provisional extracellular matrix (ECM) (Tnc, Postn, Spon2, Thbs2) as a key lung morphogenetic event associating with eosinophils. Tenascin-C (TNC) was identified as one of the key ECM markers in the lung epithelial-mesenchymal interface both at the RNA and protein levels, consistently associating with eosinophils in development and disease in mice and humans. As determined by RNA-seq analysis, naïve murine eosinophils cultured with ECM enriched in TNC significantly induced expression of Siglec-F, CD11c, eosinophil peroxidase, and other markers typical for activated eosinophils in development and allergic inflammatory responses. TNC knockout mice had an altered eosinophil recruitment profile in development. Collectively, our results indicate that lung morphogenetic processes associated with heightened Type 2 immunity are not merely a tissue "background" but specifically guide immune cells both in development and pathology.

The Impact of Climate Change on Metal Transport in a Lowland Catchment.

This study investigates the impact of future climate change on heavy metal (i.e., Cd and Zn) transport from soils to surface waters in a contaminated lowland catchment. The WALRUS hydrological model is employed in a semi-distributed manner to simulate current and future hydrological fluxes in the Dommel catchment in the Netherlands. The model is forced with climate change projections and the simulated fluxes are used as input to a metal transport model that simulates heavy metal concentrations and loads in quickflow and baseflow pathways. Metal transport is simulated under baseline climate ("2000-2010") and future climate ("2090-2099") conditions including scenarios for no climate change and climate change. The outcomes show an increase in Cd and Zn loads and the mean flux-weighted Cd and Zn concentrations in the discharged runoff, which is attributed to breakthrough of heavy metals from the soil system. Due to climate change, runoff enhances and leaching is accelerated, resulting in enhanced Cd and Zn loads. Mean flux-weighted concentrations in the discharged runoff increase during early summer and decrease during late summer and early autumn under the most extreme scenario of climate change. The results of this study provide improved understanding on the processes responsible for future changes in heavy metal contamination in lowland catchments.

A novel approach in calculating site-specific aftercare completion criteria for landfills in The Netherlands: Policy developments.

As part of a more circular economy, current attention on waste is shifting from landfilling towards the prevention, re-use and recycling of waste materials. Although the need for landfills is decreasing, there are many landfills around the world that are still operational or at the point of starting the aftercare period. With traditional aftercare management, these landfills require perpetual aftercare at considerable cost due to monitoring and regular maintenance of liners. In an attempt to lower these aftercare costs, and to prevent that future generations become responsible for finding a sustainable solution of present day waste, the Dutch government takes action to explore the possibilities of sustainable landfill management. A project was started to investigate whether the use of source-oriented treatment techniques (so-called active treatment) of landfills can result in a sustainable emission reduction to soil and groundwater. During the next decade, sustainable landfill management is tested at three selected pilot landfills in the Netherlands. To enable this pilot testing and to determine its success after the experimental treatment period, a new methodology and conceptual framework was developed. The aim of this paper is to describe the development of the new methodology, and in particular the policy decisions, needed to determine whether the pilot experiments will be successful. The pilot projects are considered successful when the concentrations in the leachate of the pilot landfills have sufficiently been reduced and for longer periods of time and comply with the derived site-specific Environmental Protection Criteria (EPC). In that case, aftercare can be reduced, and it can be determined whether sustainable landfill management is economically feasible for further implementation.

A novel screening system for secretion of heterologous proteins in Bacillus subtilis.

High-level production of secretory proteins in Bacillus subtilis leads to a stress response involving the two-component system CssRS and its target genes htrA and htrB. Here, we used this sensing system in a reporter strain in which gfp is under control of P(htrA) , the secretion stress responsive promoter of htrA. Overexpression of heterologous secretory proteins in this strain results in green fluorescent cells, which can be separated from non-secreting, low fluorescent cells using a fluorescence-activated cell sorter (FACS). Using this principle, genomic libraries of uncharacterized prokaryotic organisms, expressed in the reporter strain, can be screened for genes encoding secretory proteins.

Gene expression array testing of FFPE archival breast tumor samples: an optimized protocol for WG-DASL sample preparation.

Archived formalin-fixed, paraffin embedded (FFPE) tissues constitute a vast, well-annotated, but underexploited resource for the molecular study of cancer progression, largely because degradation, chemical modification, and cross-linking, render FFPE RNA a suboptimal substrate for conventional analytical methods. We report here a modified protocol for RNA extraction from FFPE tissues which maximized the success rate (with 100% of samples) in the expression profiling of a set of 60 breast cancer samples on the WG-DASL platform; yielding data of sufficient quality such that in hierarchical clustering (a) 12/12 (100%) replicates correctly identified their respective counterparts, with a high self-correlation (r = 0.979), and (b) the overall sample set grouped with high specificity into ER+ (38/40; 95%) and ER- (18/20; 90%) subtypes. These results indicate that a large fraction of decade-old FFPE samples, of diverse institutional origins and processing histories, can yield RNA suitable for gene expression profiling experiments.

Early versus late start of immunosuppressive therapy in idiopathic membranous nephropathy: a randomized controlled trial.

BACKGROUND: Immunosuppressive therapy in idiopathic membranous nephropathy (iMN) is debated. Accurate identification of patients at high risk for end-stage renal disease (ESRD) allows early start of therapy in these patients. It is unknown if early start of therapy is more effective and/or less toxic than late start (i.e. when GFR deteriorates). METHODS: We conducted a randomized open-label study in patients with iMN, a normal renal function and a high risk for ESRD (urinary beta2m >0.5 microg/min, UIgG >125 mg/day). Patients started with immunosuppressive therapy (cyclophosphamide for 12 months, and steroids) either immediately after randomization or when renal function deteriorated (DeltasCr > or =+25% and sCr >135 micromol/l or DeltasCr > or =+50%). End points were remission rates, duration of the nephrotic syndrome (NS), renal function and complications. RESULTS: The study included 26 patients (24 M/2 F), age 48 +/- 12 years; sCr 96 micromol/l (range 68-126) and median proteinuria 10.0 g/10 mmol Cr. Early treatment resulted in a more rapid onset of remission (P = 0.003) and a shorter duration of the NS (P = 0.009). However, at the end of the follow-up (72 +/- 22 m), there were no differences in overall remission rate, sCr (93 versus 105 micromol/l), proteinuria, relapse rate and adverse events. CONCLUSIONS: In high-risk patients with iMN, immunosuppressive treatment is effective in inducing a remission. Early treatment shortens the duration of the nephrotic phase, but does not result in better preservation of renal function. Our study indicates that treatment decisions must be based on risk and benefit assessment in the individual patient.

Differential patterns of allelic loss in estrogen receptor-positive infiltrating lobular and ductal breast cancer.

The two main histological types of infiltrating breast cancer, lobular (ILC) and the more common ductal (IDC) carcinoma are morphologically and clinically distinct. To assess the molecular alterations associated with these breast cancer subtypes, we conducted a whole-genome study of 166 archival estrogen receptor (ER)-positive tumors (89 IDC and 77 ILC) using the Affymetrix GeneChip(R) Mapping 10K Array to identify sites of loss of heterozygosity (LOH) that either distinguished, or were shared by, the two phenotypes. We found single nucleotide polymorphisms (SNPs) of high-frequency LOH (>50%) common to both ILC and IDC tumors predominately in 11q, 16q, and 17p. Overall, IDC had a slightly higher frequency of LOH events across the genome than ILC (fractional allelic loss = 0.186 and 0.156). By comparing the average frequency of LOH by chromosomal arm, we found IDC tumors with significantly (P < 0.05) higher frequency of LOH on 3p, 5q, 8p, 9p, 20p, and 20q than ILC tumors. We identified additional chromosomal arms differentiating the subtypes when tumors were stratified by tumor size, mitotic rate, or DNA content. Of 5,754 informative SNPs (>25% informativity), we identified 78 and 466 individual SNPs with a higher frequency of LOH (P < 0.05) in ILC and IDC tumors, respectively. Hierarchical clustering of these 544 SNPs grouped tumors into four major groups based on their patterns of LOH and retention of heterozygosity. LOH in chromosomal arms 8p and 5q was common in higher grade IDC tumors, whereas ILC and low-grade IDC grouped together by virtue of LOH in 16q.

Weekly paclitaxel in the treatment of recurrent ovarian carcinoma.

PURPOSE: To study the effect of weekly paclitaxel in the treatment of recurrent ovarian and peritoneal carcinoma. METHODS: A retrospective analysis of patients treated at Christie Cancer Centre between May 2003 and May 2005 was carried out. RESULTS: Fortynine patients with recurrent ovarian and peritoneal carcinoma were treated. The mean duration of treatment was 11 weeks, with 27 (54%) patients receiving 12 or more treatments. The most frequent non-haematological toxicities reported were mild nausea, constipation, lethargy and neuropathy. Moderate anaemia was noted in 50% of patients. Radiological assessment by CT scanning showed that complete or partial responses were achieved in 28% of patients. CA125 response was demonstrated in 63% of patients. Median time to recurrence was 149 days and median survival was 359 days. CONCLUSION: This study provides evidence for the role of weekly paclitaxel in the treatment of recurrent ovarian and peritoneal carcinoma even in a drug-resistant setting following multiple lines of prior therapy.

Dopamine receptor DRD2 genotype and smoking cessation outcome following treatment with bupropion SR.

The A1 allele of the dopamine D2 receptor gene (DRD2) is associated with a reduced number of dopamine binding sites in the brain and with the increased likelihood of substance abuse and addictive behavior. In a study of smokers enrolled in an open-label, randomized effectiveness trial, we investigated whether variants in the DRD2 receptor gene are associated with smoking cessation outcomes following treatment with a combination of bupropion SR and behavioral counseling. Adherence to treatment and point-prevalent smoking status were assessed at 3 and 12 months, respectively, following a target quit date. Compared to women who carry both A2 alleles, women with at least one A1 allele were more likely to report having stopped taking bupropion due to medication side effects (odds ratio (OR)=1.91, 95% confidence interval (CI)=1.01-3.60; P<0.04) and at 12 months were somewhat more likely to report smoking (OR=0.76, 95% CI=0.56-1.03; P<0.076). Significant associations or trends were not observed in men. In women, individual variability in responsiveness to bupropion-based treatment may be partially due to differences in genetic variants influencing dopamine receptor function.

Constructing land-use maps of the Netherlands in 2030.

The National Environmental Assessment Agency of the RIVM in the Netherlands is obliged to report on future trends in the environment and nature every 4 years. The last report, Nature Outlook 2, evaluated the effects of four alternative socio-economic and demographic scenarios on nature and the landscape. Spatially detailed land-use maps are needed to assess effects on nature and landscape. The objective of the study presented here was how to create spatially detailed land-use maps of the Netherlands in 2030 using the Environment Explorer, a Cellular Automata-based land-use model to construct land-use maps from four scenarios. One of these is discussed in great detail to show how the maps were constructed from the various scenario elements, story lines and additional data and assumptions on national, regional and local land-use developments. It was the first time in the history of our outlooks that consistent, spatially detailed land-use maps of the Netherlands for 2030 were constructed from national economic and demographic scenarios. Each map represents a direct reflection of model input and assumptions. The maps do not show the most probable developments in the Netherlands but describe the possible change in land use if Dutch society were to develop according to one of the four scenarios. The large (societal) uncertainties are reflected in the total set of future land-use maps. The application of a land-use model such as the Environment Explorer ensures that all relevant aspects of a scenario, i.e. economic and demographic developments, zoning policies and urban growth, are integrated systematically into one consistent framework.