High prevalence of early asymptomatic venous thromboembolism in anticoagulated COVID-19 patients hospitalized in general wards.

Coronavirus disease 2019 (COVID-19) is associated with coagulation activation and high incidence of venous thromboembolism (VTE) in severe patients despite routine thromboprophylaxis. Conflicting results exist regarding the epidemiology of VTE for unselected anticoagulated COVID-19 patients hospitalized in general wards. The aim of this study was to evaluate the prevalence of asymptomatic deep venous thrombosis (DVT) in unselected patients with COVID-19 recently hospitalized in general wards. We performed a systematic complete doppler ultrasound (CDU) at a median 4 days after admission in 42 consecutive COVID-19 patients hospitalized in general wards of our university hospital, irrespective of D-Dimer level, and retrospectively collected clinical, biological and outcome data from electronic charts. Thromboprophylaxis was systematically applied following a French national proposal. In our population, the prevalence of asymptomatic DVT was 19% (8/42 patients), with distal thrombosis in 7/8 cases and bilateral DVT in 4/8 cases. Symptomatic pulmonary embolism was detected in 4 (9.5%) patients, associated to DVT in one case. Compared to patients without DVT, patients with DVT were older and experienced poorer outcomes. In conclusion, prevalence of asymptomatic DVT is high in the first days of hospitalization of unselected COVID-19 patients in general wards and may be related to poor prognosis. Individualized assessment of thromboprophylaxis and early systematic screening for DVT is warranted in this context.

A surface-tethered spheroid model for functional evaluation of 3T3-L1 adipocytes.

In order to effectively treat obesity, it must be better understood at the cellular level with respect to metabolic state and environmental stress. However, current two-dimensional (2D) in vitro cell culture methods do not represent the in vivo adipose tissue appropriately due to the absence of complex architecture and cellular signaling. Conversely, 3D in vitro cultures have been reported to have optimal results mimicking the adipose tissue in vivo. The main aim of this study was to examine the efficacy of a novel conjugate of a genetically engineered polymer, elastin-like polypeptide (ELP) and a synthetic polymer, polyethyleneimine (PEI), toward creating a 3D preadipocyte culture system. We then used this 3D culture model to study the preadipocyte differentiation and adipocyte maintenance processes when subjected to various dosages of nutritionally relevant free fatty acids with respect to total DNA and protein content, cell viability, and intracellular triglyceride accumulation. Our results showed that 3T3-L1 preadipocytes cultured on the ELP-PEI surface formed 3D spheroids within 72 h, whereas the cells cultured on unmodified tissue culture polystyrene surfaces remained in monolayer configuration. Significant statistical differences were discovered between the 3D spheroid and 2D monolayer culture with respect to the DNA and protein content, fatty acid consumption, and triglyceride accumulation, indicating differences in cellular response. Results indicated that the 3D culture may be a more sensitive modeling technique for in vitro adipocyte culture and provides a platform for future evaluation of 3D in vitro adipocyte function.

Effect of processing temperature on the morphology and drug-release characteristics of elastin-like polypeptide-collagen composite coatings.

Elastin-like polypeptides (ELPs) exhibit an inverse phase transition temperature (Tt) in response to changes in their environment. We hypothesized that processing ELP-collagen composites at temperatures higher than the Tt of ELP (∼32 °C) will affect their microstructure and subsequently, achieve tunable release of model drugs. The composite coatings were prepared by formation of ELP-collagen hydrogels at 37 °C, incubation at 37, 45, or 55 °C, and finally air-drying at 37 °C. Scanning electron micrographs revealed that the fabrication process affected both the collagen and ELP microaggregate phases. A gradual time dependent bovine serum albumin (BSA) release that followed the power law and a burst antibiotic doxycycline release followed by a linear zero-order release were observed. Importantly, BSA and doxycycline releases were dependent on the ELP microaggregate size, which was governed by the processing temperatures. This study lays the foundation to achieve optimized composite microstructures by controlling processing conditions for drug delivery applications.

Drugs detected in suspected pediatric ingestions: a three-year review.

Comprehensive toxicology testing can be a valuable resource when pediatric patients are suspected of having ingested unknown chemicals. In a recent 36-month period (January 2000-December 2002), the Analytical Toxicology Laboratory at the University of Mississippi Medical Center, Jackson, MS, tested 190 urine samples referred from the hospital's Pediatric Emergency Department for STAT toxicology testing. All samples were tested in parallel using an immunoassay technique for drugs of abuse and a comprehensive technique using gas chromatography/mass spectroscopy (GC/MS). With immunoassay, sixteen percent of the samples were positive for a drug of abuse. In comparison, drugs were detected in 85% of the samples using comprehensive GC/MS analysis. A total of 86 different pharmaceuticals were identified by GC/MS. This review shows that comprehensive toxicology testing provides the clinician significantly more information regarding the exposure of the pediatric patient presenting with an unknown chemical ingestion.

Vitamin D Supplementation Suppresses Hypoxia-Stimulated Placental Cytokine Secretion, Hypertension and CD4+ T Cell Stimulation in Response to Placental Ischemia.

OBJECTIVE: To investigate a role of Vitamin D in the pathogenesis of preeclampsia (PE), and to discern any potential benefits of Vitamin D supplementation on hypertension in the RUPP rat model of PE. STUDY DESIGN: Blood and placentas from normal pregnancies (NP) and PE were collected following elective cesarean delivery without evidence of infection. Circulating Vitamin D was extracted by HPLC and measured via mass spectrometry. Media for placenta explants was supplemented with Vitamin D and exposed to hypoxic (1% O2) or normoxic (6% O2) conditions for 24 hours. ELISAs were performed on media and normalized to total protein to determine cytokine secretion. RUPP rats were supplemented with vitamin D by oral gavage, and blood pressure (MAP) and pup weights were measured in NP and RUPP rats with or without Vitamin D supplementation. Flow cytometry was used to evaluate CD4+ Tcells in control RUPP rats and RUPP rats treated with Vitamin D. RESULTS: Inflammatory cytokine secretion was higher (p<0.05) while the anti-inflammatory cytokine, IL-10, was significantly lower in the media of PE placentas compared to NP (p=0.005). Vitamin D supplementation decreased hypoxia stimulated pro-inflammatory cytokine secretion (p=0.003) in the media of PE placentas. Vitamin D decreased MAP and circulating CD4+ T cells in the RUPP rat model of PE (p<0.05). CONCLUSION: Vitamin D supplementation may be useful in the treatment or prevention of hypertensive disorders in pregnancy.

Determination of derivatized urea in exhaled breath condensate by LC-MS.

Elevation in one or more compounds in exhaled breath condensate (EBC) has been reported to be related to one or another lung disease. The increased concentration might be caused by increased chemicals in the airway surface liquid. However, it might also be due to an increased delivery of liquid samples into the airstream. Being evenly distributed throughout the body, urea is a likely candidate for a marker of such dilution. A liquid chromatography-tandem mass spectrometry method was developed for determination of EBC urea. Urea in EBC samples was converted to 2-hydroxypyrimidine (2-HPM) through a one step reaction, along with (15)N(2)-urea added as an internal standard. The product ion m/z 97/42 was selected for quantification with m/z 99/43 from (15)N(2)-2-HPM as a standard. Concentrations of urea in EBC from five lung cancer patients were found to be 35.1, 2.2, 103.5, 19.3, and 3.6 microM, respectively. The highest values were in patients dying of respiratory distress, whose lungs were filled with fluid. Lower values were seen in patients whose conditions were improving. Lately, one of the low EBC urea values was observed in a patient whose airway status did not contribute to his poor clinical condition.

Suspected pediatric ingestions: effectiveness of immunoassay screens vs. gas chromatography/mass spectroscopy in the detection of drugs and chemicals.

Rapid and accurate analytical testing can be of great value when determining treatment for pediatric patients suspected of ingesting an unknown chemical. Though often overlooked, gas chromatography/mass spectroscopy (GC/MS) can be a valuable resource in emergency toxicology testing. In a recent 24-month period (July 1999-June 2001), the Analytical Toxicology Laboratory at the University of Mississippi Medical Center, Jackson, MS, compared the results of GC/MS analysis to results obtained by immunoassay testing. The laboratory tested 139 urine samples referred for STAT toxicology testing from the hospital's Pediatric Emergency Department. All samples were tested in parallel using an immunoassay technique (EMIT) and GC/MS. With analysis by immunoassay, 17.3% of the samples were positive for a drug of abuse. The number of positive drug classes ranged from 0 to 2 per sample (mean 0.17 +/- 0.43) using immunoassay. With analysis by GC/MS, drugs were detected in 88.5% of the samples. The number of drugs detected ranged from 0 to 11 per sample (mean 2.2 +/- 1.8) with GC/MS. A total of 64 different pharmaceuticals were identified by GC/MS. This study shows that analysis by GC/MS offers the clinician a more comprehensive view into the exposure of the pediatric patient presenting with an unknown chemical ingestion.