Perioperative Delta Weight and Pediatric Obstructive Sleep Apnea Resolution after Adenotonsillectomy.

OBJECTIVE(S): The first-line treatment for pediatric obstructive sleep apnea (OSA) is adenotonsillectomy. Post-operative weight gain is a well-documented phenomenon. We hypothesized that higher peri-adenotonsillectomy delta weight correlates with lower rates of OSA resolution in pediatric patients. METHODS: This was a retrospective cohort study consisting of 250 patients from 2 to 17 years of age at a tertiary academic medical center between January 2021 and December 2022. Polysomnography results and body mass index (BMI) changes were collected through the electronic health record. Univariate and multivariate logistical regression analyses were performed, adjusting for confounding factors. RESULTS: Perioperative delta weight and pre-operative baseline AHI values were significant predictors of residual OSA. For every 1-kilogram gain in weight, the odds of residual OSA (AHI >5) increase by 6.0% (OR = 1.06, 95% CI = 1.02-1.10, p < 0.002), and the odds of residual severe OSA (AHI > 10) increase by 8% (OR = 1.08, 95% CI = 1.04-1.12, p < 0.001). Increased AHI, Black/African American race, and male sex were also factors associated with incomplete OSA resolution. CONCLUSIONS: Increased peri-adenotonsillectomy delta weight is associated with higher rates of residual OSA in children. Patients and families should be counseled about appropriate weight loss and control methods before adenotonsillectomy. LEVEL OF EVIDENCE: IV Laryngoscope, 134:4141-4147, 2024.

Adopting the Swenson-like technique for patients with Hirschsprung disease in Vietnam.

PURPOSE: To evaluate outcomes of adopting the Swenson-like technique in transanal pull-through for Hirschsprung disease (HD) at a children's institution in Southeast Asia. METHODS: A retrospective chart review was performed over 42 months at a safety-net pediatric hospital in Vietnam. RESULTS: From January 2019 to June 2022, the Swenson-like technique was implemented and performed on 139 patients (115 male, 24 female, mean age 6.4 ± 11.96 months). There were 123 transanal-only resections, 5 transanal plus laparoscopic, and 11 transanal plus laparotomy. The mean operative time was 70.9 ± 43.6 min. The average length of the resected specimen was 18.8 ± 10.9 cm. No urethral or vaginal injuries occurred. Postoperative complications included 1 anastomotic leak (0.7%), 4 anastomotic stenoses (2.8%), and 2 perianal abscesses in the setting of recurrent enterocolitis (1.4%). With a mean follow-up of 26 ± 11.9 months (range 3-48 months), there were 25 enterocolitis cases (17.9%), 45 patients with constipation (32.3%), 1 episode of fecal incontinence lasting longer than 6 months (0.7%), and 1 rectal-vestibular fistula noted one year postoperatively (0.7%). There were no deaths. CONCLUSION: The Swenson-like technique in transanal pull-through for Hirschsprung's disease was safely adopted at our institution. The results show that applying this technique uniformly in a single institution led to excellent outcomes.

Human clinical mutations in mitochondrially encoded subunits of Complex I can be successfully modeled in E. coli.

Respiratory Complex I is the site of a large fraction of the mutations that appear to cause mitochondrial disease. Seven of its subunits are mitochondrially encoded, and therefore, such mutants are particularly difficult to construct in cell-culture model systems. We have selected 13 human clinical mutations found in ND2, ND3, ND4, ND4L, ND5 and ND6 that are generally found at subunit interfaces, and not in critical residues. These mutations have been modeled in E. coli subunits of Complex I, nuoN, nuoA, nuoM, nuoK, nuoL, and nuoJ, respectively. All mutants were expressed from a plasmid encoding the entire nuo operon, and membrane vesicles were analyzed for deamino-NADH oxidase activity, and proton translocation activity. ND5 mutants were also analyzed using a time-delayed expression system, recently described by this lab. Other mutants were analyzed for the ability to associate in subcomplexes, after expression of subsets of the genes. For most mutants there was a positive correlation between those that were previously determined to be pathogenic, or likely to be pathogenic, and those that we found with compromised Complex I activity or subunit interactions in E. coli. In conclusion, this approach provides another way to explore the deleterious effects of human mitochondrial mutations, and it can contribute to molecular understanding of such mutations.

Characteristics, incidence, and trends of intimate partner homicides in Massachusetts: Patterns by birthplace, race, and ethnicity.

This study compared the incidence rates of intimate partner homicide (IPH) in Massachusetts by place of birth and race/ethnicity. The analysis involved 340 IPH victim cases between 1994 and 2014. Victims were just under 40 years of age, on average, and most were female (85%), White (67%), and killed by stabbing (34.4%) or firearms (33%). The incidence of IPH victims ranged from 1.3 to 5.6 cases per million people per year between 1994 and 2014 (M = 2.4 per million). Foreign-born individuals had 1.9-fold higher IPH incidence rates of victims relative to U.S.-born individuals. The incidence of IPH-suicide victims was also significantly higher among foreign-born (M = 1.2 per million) relative to U.S.-born individuals (M = 0.4 per million). Furthermore, minority racial/ethnic groups had significantly high incidence rates of victims, with highest incidence of IPH among Blacks. In the full sample there was a 1.9% decline in the incidence of IPH victims per year, which was not statistically significant. The findings highlight the need for culturally specific prevention and intervention strategies to address risks of IPHs and IPH-suicides among diverse groups, particularly among groups most at-risk in Massachusetts such as foreign-born born individuals and racial and ethnic minorities.

Analysis of Human Mutations in the Supernumerary Subunits of Complex I.

Complex I is the largest member of the electron transport chain in human mitochondria. It comprises 45 subunits and requires at least 15 assembly factors. The subunits can be divided into 14 "core" subunits that carry out oxidation-reduction reactions and proton translocation, as well as 31 additional supernumerary (or accessory) subunits whose functions are less well known. Diminished levels of complex I activity are seen in many mitochondrial disease states. This review seeks to tabulate mutations in the supernumerary subunits of humans that appear to cause disease. Mutations in 20 of the supernumerary subunits have been identified. The mutations were analyzed in light of the tertiary and quaternary structure of human complex I (PDB id = 5xtd). Mutations were found that might disrupt the folding of that subunit or that would weaken binding to another subunit. In some cases, it appeared that no protein was made or, at least, could not be detected. A very common outcome is the lack of assembly of complex I when supernumerary subunits are mutated or missing. We suggest that poor assembly is the result of disrupting the large network of subunit interactions that the supernumerary subunits typically engage in.

Cytokine-induced molecular responses in airway smooth muscle cells inform genome-wide association studies of asthma.

BACKGROUND: A challenge in the post-GWAS era is to assign function to disease-associated variants. However, available resources do not include all tissues or environmental exposures that are relevant to all diseases. For example, exaggerated bronchoconstriction of airway smooth muscle cells (ASMCs) defines airway hyperresponsiveness (AHR), a cardinal feature of asthma. However, the contribution of ASMC to genetic and genomic studies has largely been overlooked. Our study aimed to address the gap in data availability from a critical tissue in genomic studies of asthma. METHODS: We developed a cell model of AHR to discover variants associated with transcriptional, epigenetic, and cellular responses to two AHR promoting cytokines, IL-13 and IL-17A, and performed a GWAS of bronchial responsiveness (BRI) in humans. RESULTS: Our study revealed significant response differences between ASMCs from asthma cases and controls, including genes implicated in asthma susceptibility. We defined molecular quantitative trait loci (QTLs) for expression (eQTLs) and methylation (meQTLs), and cellular QTLs for contractility (coQTLs) and performed a GWAS of BRI in human subjects. Variants in asthma GWAS were significantly enriched for ASM QTLs and BRI-associated SNPs, and near genes enriched for ASM function, many with small P values that did not reach stringent thresholds of significance in GWAS. CONCLUSIONS: Our study identified significant differences between ASMCs from asthma cases and controls, potentially reflecting trained tolerance in these cells, as well as a set of variants, overlooked in previous GWAS, which reflect the AHR component of asthma.

Ten new species of Calonectria from Indonesia and Vietnam.

Vietnam and Indonesia have rapidly growing and extensive plantation forestry programs, especially of Acacia spp. and Eucalyptus spp. As these plantations expand, the threat from pests and diseases also increases. Calonectria species are among those pathogens causing diseases of trees in plantations and nurseries in these countries. Extensive surveys were conducted across plantations and nurseries of Vietnam and parts of Indonesia, where a large number of Calonectria isolates were retrieved from diseased leaves and soils associated with symptomatic trees. The aim of this study was to identify and resolve the phylogenetic relationships among these isolates using DNA sequence comparisons of four gene regions as well as morphological characters. From a collection of 165 isolates, the study revealed five known and 10 undescribed species. The relatively high diversity of Calonectria species found in this study supports the view that many more species in this genus remain to be discovered in other areas of Southeast Asia.

Traction Force Screening Enabled by Compliant PDMS Elastomers.

Actomyosin contractility is an essential element of many aspects of cellular biology and manifests as traction forces that cells exert on their surroundings. The central role of these forces makes them a novel principal therapeutic target in diverse diseases. This requires accurate and higher-capacity measurements of traction forces; however, existing methods are largely low throughput, limiting their utility in broader applications. To address this need, we employ Fourier-transform traction force microscopy in a parallelized 96-well format, which we refer to as contractile force screening. Critically, rather than the frequently employed hydrogel polyacrylamide, we fabricate these plates using polydimethylsiloxane rubber. Key to this approach is that the polydimethylsiloxane used is very compliant, with a lower-bound Young's modulus of ∼0.4 kPa. We subdivide these monolithic substrates spatially into biochemically independent wells, creating a uniform multiwell platform for traction force screening. We demonstrate the utility and versatility of this platform by quantifying the compound and dose-dependent contractility responses of human airway smooth muscle cells and retinal pigment epithelial cells. By directly quantifying the endpoint of therapeutic intent, airway-smooth-muscle contractile force, this approach fills an important methodological void in current screening approaches for bronchodilator drug discovery, and, more generally, in measuring contractile response for a broad range of cell types and pathologies.

Synthesis, resolution and crystal structures of two enanti-omeric rhodamine derivatives.

The title mol-ecule, rac-6'-bromo-3'-di-ethyl-amino-3H-spiro-[2-benzo-furan-1,9'-xanthen]-3-one, C24H20BrNO3, was synthesized and the two enanti-omers which formed were separated. The structures of all three compounds were determined and compared with those of a variety of related derivatives. A notable feature is the fold of the xanthene portion which ranges from 15.15 (13)° in the racemate to 2.42 (2)° in one mol-ecule of the R enanti-omer with that for the S enanti-omer having an inter-mediate value. The differences are attributed to the number and severity of inter-molecular inter-actions which include C-H⋯O hydrogen bonds, C-H⋯π(ring) and, in the S enanti-omer, a π-stacking inter-action between the carbonyl group and an aromatic ring.

Gelsolin is an endogenous inhibitor of syncytiotrophoblast extracellular vesicle shedding in pregnancy.

BACKGROUND: Preeclampsia, a pregnancy-specific inflammatory disorder, is characterized by high levels of anti-angiogenic protein, soluble fms-like tyrosine kinase 1 (sFlt1), in the maternal circulation. sFlt1 producing molecular machinery is present in syncytiotrophoblast extracellular vesicles that are released by the placenta into maternal plasma during normal pregnancy, a process greatly accelerated in preeclampsia. We hypothesized that syncytiotrophoblast extracellular vesicles exposes cytoplasmic actin to plasma resulting in depletion of plasma gelsolin (pGSN), an abundant plasma protein that scavenges circulating actin and other pro-inflammatory mediators. OBJECTIVE: To test whether pGSN levels would be lower in preeclampsia and to assess whether recombinant human plasma gelsolin (rhpGSN) may promote placental health by decreasing shedding of syncytiotrophoblast extracellular vesicles. METHODS: We tested pGSN levels in third trimester plasma samples from women with preeclampsia and non-hypertensive pregnancies. We then assessed whether rhpGSN may act as a negative regulator of syncytial shedding in placental explant culture and dynamic mechanical stretch studies. RESULTS: pGSN levels fall in late pregnancy and decline further in preeclampsia patients. Recombinant human pGSN (rhpGSN) at 100µg/ml limits spontaneous syncytiotrophoblast vesicle release and sFlt1 protein dissemination by normal placental explants. Higher rhpGSN doses (500µg/ml) also limit syncytiotrophoblast vesicle and sFlt1 dissemination from preeclamptic placental explants. rhpGSN also mitigates syncytiotrophoblast vesicle during dynamic mechanical stretch. CONCLUSIONS: 1) pGSN, an anti-inflammatory factor of maternal origin is reduced in preeclampsia and may contribute to disease progression and 2) exogenous rhpGSN supplementation can limit the dissemination of toxic syncytiotrophoblast vesicle that characterizes the disease state.

Dynamics of cell area and force during spreading.

Experiments on human pulmonary artery endothelial cells are presented to show that cell area and the force exerted on a substrate increase simultaneously, but with different rates during spreading; rapid-force increase systematically occurred several minutes past initial spreading. We examine this theoretically and present three complementary mechanisms that may accompany the development of lamellar stress during spreading and underlie the observed behavior. These include: 1), the dynamics of cytoskeleton assembly at the cell basis; 2), the strengthening of acto-myosin forces in response to the generated lamellar stresses; and 3), the passive strain-stiffening of the cytoskeleton.

Altered mechanobiology of Schlemm's canal endothelial cells in glaucoma.

Increased flow resistance is responsible for the elevated intraocular pressure characteristic of glaucoma, but the cause of this resistance increase is not known. We tested the hypothesis that altered biomechanical behavior of Schlemm's canal (SC) cells contributes to this dysfunction. We used atomic force microscopy, optical magnetic twisting cytometry, and a unique cell perfusion apparatus to examine cultured endothelial cells isolated from the inner wall of SC of healthy and glaucomatous human eyes. Here we establish the existence of a reduced tendency for pore formation in the glaucomatous SC cell--likely accounting for increased outflow resistance--that positively correlates with elevated subcortical cell stiffness, along with an enhanced sensitivity to the mechanical microenvironment including altered expression of several key genes, particularly connective tissue growth factor. Rather than being seen as a simple mechanical barrier to filtration, the endothelium of SC is seen instead as a dynamic material whose response to mechanical strain leads to pore formation and thereby modulates the resistance to aqueous humor outflow. In the glaucomatous eye, this process becomes impaired. Together, these observations support the idea of SC cell stiffness--and its biomechanical effects on pore formation--as a therapeutic target in glaucoma.

Active mechanics and dynamics of cell spreading on elastic substrates.

The spreading area of cells has been shown to play a central role in the determination of cell fate and tissue morphogenesis; however, a clear understanding of how spread cell area is determined is still lacking. The observation that cell area and force generally increase with substrate rigidity suggests that cell area is dictated mechanically, by means of a force-balance between the cell and the substrate. A simple mechanical model, corroborated by experimental measurements of cell area and force is presented to analyze the temporal force balance between the cell and the substrate during spreading. The cell is modeled as a thin elastic disc that is actively pulled by lamellipodia protrusions at the cell front. The essential molecular mechanisms of the motor activity at the cell front, including, actin polymerization, adhesion kinetics, and the actin retrograde flow, are accounted for and used to predict the dynamics of cell spreading on elastic substrates; simple, closed-form expressions for the evolution of cell size and force are derived. Time-resolved, traction force microscopy, combined with measurements of cell area are performed to investigate the simultaneous variations of cell size and force. We find that cell area and force increase simultaneously during spreading but the force develops with an apparent delay relative to the increase in cell area. We demonstrate that this may reflect the strain-stiffening property of the cytoskeleton. We further demonstrate that the radial cell force is a concave function of spreading speed and that this may reflect the strengthening of cell-substrate adhesions during spreading.

Assessing the impact of engineered nanoparticles on wound healing using a novel in vitro bioassay.

AIM: As engineered nanoparticles (ENPs) increasingly enter consumer products, humans become increasingly exposed. The first line of defense against ENPs is the epithelium, the integrity of which can be compromised by wounds induced by trauma, infection, or surgery, but the implications of ENPs on wound healing are poorly understood. MATERIALS & METHODS: Herein, we developed an in vitro assay to assess the impact of ENPs on the wound healing of cells from human cornea. RESULTS & DISCUSSION: We show that industrially relevant ENPs impeded wound healing and cellular migration in a manner dependent on the composition, dose and size of the ENPs as well as cell type. CuO and ZnO ENPs impeded both viability and wound healing for both fibroblasts and epithelial cells. Carboxylated polystyrene ENPs retarded wound healing of corneal fibroblasts without affecting viability. CONCLUSION: Our results highlight the impact of ENPs on cellular wound healing and provide useful tools for studying the physiological impact of ENPs.

Rates of femicide in women of different races, ethnicities, and places of birth: Massachusetts, 1993-2007.

To describe the epidemiology of intimate partner violence (IPV) homicide in Massachusetts, an IPV mortality data set developed by the Massachusetts Department of Public Health was analyzed. The rates of death were estimated by dividing the number of decedents over the aged-matched population and Poisson regression was used to estimate the contribution of race, ethnicity, and foreign-born status to the risk of dying from IPV. Out of the total 270 women whose deaths were associated with IPV, 239 (89%) were killed by a male partner. Black women had a risk of dying from IPV of 16.2 per 1,000,000 person-years. Hispanic women also had a higher risk of dying from IPV than non-Hispanic women; incidence risk ratio of 9.7 (Poisson regression 95% confidence interval 6.8-13.8). IPV femicide disproportionately affected Black and Hispanic women. Agencies must consider the importance of providing culturally appropriate services to IPV survivors and their community.

Batterer intervention program enrollment and completion among immigrant men in Massachusetts.

This study describes immigrant clients enrolled in Massachusetts batterer intervention (BI) programs from 2002 to 2004 (N = 480). Our study sought to (a) describe the immigrant men enrolled in Massachusetts BI programs, (b) investigate whether immigrants were more or less likely to complete BI programs than were nonimmigrants, and (c) investigate whether immigrants in non-English, culturally specific groups were more or less likely to complete BI programs than were immigrants in mainstream groups. Of BI program clients, 14% were immigrants. Of these, 73% were not U.S. citizens. Immigrants were more likely to complete the programs than were nonimmigrants (54% vs. 38%). Although a greater proportion of immigrants who attended non-English groups completed the programs than did immigrants who attended mainstream groups (66% and 46%), neither the bivariate nor the adjusted odds ratio was statistically significant. Possible reasons for differences between immigrant and nonimmigrant characteristics and program completion rates are discussed.