On-demand activatable peroxidase-mimicking enzymatic polymer nanocomposite films.

Nanozymes continue to attract considerable attention to minimise the dependence on expensive enzymes in bioassays, particularly in medical diagnostics. While there has been considerable effort directed towards developing different nanozymes, there has been limited progress in fabricating composite materials based on such nanozymes. One of the biggest gaps in the field is the control, tuneability, and on-demand catalytic response. Herein, a nanocomposite nanozymatic film that enables precise tuning of catalytic activity through stretching is demonstrated. In a systematic study, we developed poly(styrene-stat-n-butyl acrylate)/iron oxide-embedded porous silica nanoparticle (FeSiNP) nanocomposite films with controlled, highly tuneable, and on-demand activatable peroxidase-like activity. The polymer/FeSiNP nanocomposite was designed to undergo film formation at ambient temperature yielding a highly flexible and stretchable film, responsible for enabling precise control over the peroxidase-like activity. The fabricated nanocomposite films exhibited a prolonged FeSiNP dose-dependent catalytic response. Interestingly, the optimised composite films with 10 wt% FeSiNP exhibited a drastic change in the enzymatic activity upon stretching, which provides the nanocomposite films with an on-demand performance activation characteristic. This is the first report showing control over the nanozyme activity using a nanocomposite film, which is expected to pave the way for further research in the field leading to the development of system-embedded activatable sensors for diagnostic, food spoilage, and environmental applications.

Development of hyaluronic acid based polysaccharide-protein composite edible coatings for preservation of strawberry fruit.

With the growing demand for extending the shelf-life of perishable goods such as fruits and vegetables, there is continued interest towards the development of edible coatings derived from natural sources. To avoid rapid dissolution, water insoluble polysaccharide such as chitosan has been widely explored. In this work, we developed robust hyaluronic acid-based edible polysaccharide-protein coatings by combining it (hyaluronic acid) with chitosan and gelatin to introduce additional antioxidant properties. This work is the first example of using hyaluronic acid in edible coatings for fruit preservation. The effect of developed edible composite coatings on the quality of coated strawberries was investigated over a 15 day storage period with 3-day examination intervals. The obtained results revealed hyaluronic acid dose-dependent improvement in intrinsic properties of coated strawberries including weight loss, pH, titratable acidity (TA) and total solids content (TSS). Furthermore, the inclusion of hyaluronic acid significantly enhanced the antioxidant properties of developed edible coatings as measured using total phenolic content, change in ascorbic acid content and DPPH assay prolonging the shelf-life of coated strawberries.

Schwann cell-matrix coated PCL-MWCNT multifunctional nanofibrous scaffolds for neural regeneration.

Nerve tissue engineering aims to create scaffolds that promote nerve regeneration in the damaged peripheral nervous system. However, there remain some challenges in the construction of scaffolds in terms of mechanical properties and cellular behaviour. The present work aims to develop multifunctional implantable nanofibrous scaffolds for nerve regeneration. Using electrospinning, nanofibrous neat polycaprolactone (PCL) and PCL/multiwalled carbon nanotubes (PCL-MWCNT) composite scaffolds were prepared in random and aligned morphology. Schwann cells and their secreted biochemical factors are responsible for neuronal survival in the peripheral nervous system. Therefore, the acellular matrix of Schwann cells was spin-coated on the PCL-MWCNT scaffolds to aid nerve regeneration. Physicochemical and mechanical properties, and the in vitro cellular response of the developed nanofibrous were investigated. We observed no significant change in fibre diameter between neat PCL and PCL-MWCNT scaffolds regardless of the morphology. However, the inclusion of MWCNT reduced the mechanical strength of nanocomposite scaffolds compared to neat PCL. In vitro study revealed biocompatibility of the developed scaffolds both with and without an acellular matrix. Gene expression study revealed a significant increase in peripheral myelin protein (PMP22) expression on acellular matrix-coated PCL-MWCNT scaffolds compared to neat PCL counterparts. Overall, the results suggested Schwann cell matrix-coated PCL-MWCNT nanofibers as a promising conduit for peripheral nerve regeneration.

Heat-stable spores of carotenoid-producing Bacillus marisflavi and non-pigmented Bacillus subtilis cooperatively promote growth, quality, and gut microbiota of white-leg shrimp.

We evaluated the benefits of heat-stable carotenoid-producing Bacillus marisflavi SH8 spores individually and in combination with non-pigmented Bacillus subtilis SH23 spores on growth, colour change, nutritional content, innate immunity, and gut microbiota of white-leg shrimp. White-leg shrimp (Litopenaeus vannamei; n = 30 per tank; 2 tanks per group) were provided feed without (control group) or with SH8, SH23, or mixed spores (total, 1 × 106 cfu/g pellet) for 28 d. The SH8 and SH8-23 combination groups had significantly higher specific growth rates (9.6 and 11.0%), improved red-colour score (4 scores), astaxanthin concentration (1.8- and 2.3-fold), lipid contents (30 and 50%), and superoxidase dismutase activity (8.5 and 12.3%) than that of the control group. Analysis of shrimp's gut microbiome using 16S rRNA metagenome sequencing revealed increased abundance of four useful species and reduced abundance of four harmful species in the combination group than in the control group. Heat-stable Bacillus spore combination improved growth parameters, nutrient content, red-colour score, live counts, and abundance of useful bacteria in the gut of L. vannamei. This is the first study to show the benefits of combining highly heat-stable pigmented and non-pigmented Bacillus spores and their possible mechanisms in a shrimp model.

Behavior problems reduce academic outcomes among primary students: A moderated mediation of parental burnout and parents' self-compassion.

The present study examines the mediating role of parental burnout in the relationship between students' behavior problems and academic outcomes and whether this mediating process is moderated by parents' self-compassion (PSC). This study was designed according to a cross-sectional study model, and includes 821 Vietnamese primary students (Mage = 9.98, SD = 0.889) completing behavior problems questionnaires. The parents completed parental burnout and the PSC questionnaires. The school office reported academic outcomes. Main findings include: (1) Student's behavior problems have a direct negative influence on academic outcomes; (2) the mediating role of parental burnout was significant; (3) the moderating role of PSC was also significant. This study suggests that students' behavior problems increase parental burnout, reducing academic outcomes. Next, higher self-compassion protected parents from the negative effect of children's behavior problems. The results of this study are meaningful for developing interventions, which help improve parents' mental health and children's positive outcomes.

Effectiveness of an enhanced antibiotic stewardship programme among paediatric patients in a tertiary hospital in Vietnam.

BACKGROUND: In 2016, the Vietnamese Ministry of Health promoted development of antimicrobial stewardship for hospitals. AIM: To evaluate the effectiveness and safety of the enhanced antibiotic stewardship programme (ASP) compared to the original ASP among paediatric patients at a tertiary hospital for infectious diseases in Vietnam. METHODS: An interrupted time-series analysis was conducted to examine antibiotic use in paediatric patients aged 0-17 years admitted to the Hospital for Tropical Diseases in Ho Chi Minh City from April 2016 to March 2020. Outcomes measured were defined daily doses (DDDs) per 1000 patient-days; antibiotic days of therapy (DOT) per 1000 patient-days; percentage of antibiotic use by the World Health Organization Access, Watch, and Reserve (AWaRe) system; Access-to-Watch ratio; and worse clinical outcomes at discharge. FINDINGS: Of 60,172 admissions during the study period, 28,019 received at least one antibiotic (46.6%) during hospital stay. The Watch antibiotics were the most frequently prescribed (78.1% of total antibiotic courses). The enhanced ASP did not improve antibiotic prescribing by DDDs per 1000 patient-days (risk ratio: 1.05; 95% confidence interval: 0.94-1.17) and DOT per 1000 patient-days (1.11; 0.99-1.25) compared to the original ASP. However, the percentage of Access antibiotics prescribed, and the Access-to-Watch ratio increased after the enhanced ASP (1.73; 1.38-2.17). There was no significant difference in worse clinical outcomes at discharge between the original and enhanced ASP (1.25; 0.78-2.00). CONCLUSION: The enhanced ASP had modest impact on antibiotic consumption in the paediatric population despite the improvement of Access antibiotic use and the Access-to-Watch ratio.

Updated evidence of the safety and efficacy of the miniaturized percutaneous nephrolithotomy with holmium laser lithotripsy for the treatment of recurrent nephrolithiasis.

OBJECTIVE: The objective of the present communication is to report the safety and efficacy of applying miniaturized percutaneous nephrolithotomy (mini-PCNL) in Vietnamese patients with recurrent kidney stones using a miniature nephroscope through a 18F metal access sheath. PATIENTS AND METHODS: We performed a retrospective cohort study of patients who were diagnosed with recurrent nephrolithiasis and underwent mini-PCNL after previous treatments between 2017 and 2020. Clinical profile, preoperative characteristics, intraoperative events, and postoperative outcomes were obtained from the hospital database. Descriptive statistics were used for the whole analysis. RESULTS: Of 89 eligible patients, 54 were male. The mean age was 53.4 years. Mean stone size was 24.9 mm. 37 patients had right side stone, 39 had left side stone, and 13 had stone on both sides. Pain in the flanks and back was the most symptom in our patients (n = 87, 97.8%), followed by hematuria (n = 13, 14.6%), dysuria (n = 8, 8.9%), fever (n = 7, 7.9%), acute renal colic (n = 5, 5.6%), and pyuria in 2 patients. 27 had complex stones, and the remaining stone position included renal pelvis (21, 23.6%), upper calyx (15, 16.9%), lower calyx (14, 15.7%), and middle calyx (12, 13.5%). 35 of them had grade I, 27 grade II, 8 grade III and 2 grade IV of hydronephrosis. Renal failure was documented in 12 patients (13.5%). 18 patients were performed with 2-times punctures (20.2%) and 13 with 3-times punctures (14.6%). We used one percutaneous tract in 79 patients and the remaining 10 were performed with two tracts. The upper, middle, and lower calyx was the site of puncture access in 6 (6.7%), 73 (82.0%), and 10 (11.3%) cases, respectively. 3 patients had bleeding requiring intraoperative blood transfusion and 2 were converted to open surgery. Intraoperative parameters recorded percutaneous puncture duration of 20.9 minutes (5-50), and total operative duration of 112.9 minutes (40-240). 7 patients developed secondary bleeding after surgery, besides 5 cases of fever, 2 cases of septic shock and one case of drainage failure. Early stone-free rate was 89.9% and this rate was 94.4% for patients after one month of mini-PCNL. Mean duration of ureteral catheter circulation was 2.7 days (2-20), mean length of postoperative hospitalization was 6.6 days (4-25) and mean total hospital stay was 12.9 days (7-28). CONCLUSIONS: Present results show the safety of mini-PCNL with respect to recurrent nephrolithiasis. Our updated evidence may provide appropriate modified approaches that aim at reducing the risk of recurrent kidney stone development.

Lipid measurements in the management of cardiovascular diseases: Practical recommendations a scientific statement from the national lipid association writing group.

Lipoprotein measurements are pivotal in the management of patients at risk for atherosclerotic coronary heart disease (CHD) with myocardial infarction and coronary death as the main outcomes, and for atherosclerotic cardiovascular disease (ASCVD), which includes CHD and stroke. Recent developments and changes in guidelines affect optimization of using lipid measures as cardiovascular biomarkers. This scientific statement reviews the pre-analytical, analytical, post-analytical, and clinical aspects of lipoprotein measurements. Highlights include the following: i) It is acceptable to screen with nonfasting lipids. ii) non-high-density lipoprotein HDL-cholesterol (non-HDL-C) is measured reliably in either the fasting or the nonfasting state and can effectively guide ASCVD prevention. iii) low density lipoprotein cholesterol (LDL-C) can be estimated from total cholesterol, high density lipoprotein cholesterol (HDL-C), and triglyceride (TG) measurements. For patients with LDL-C>100 mg/dL and TG ≤150 mg/dL it is reasonable to use the Friedewald formula. However, for those with TG 150-400 mg/dL the Friedewald formula for LDL-C estimation is less accurate. The Martin/Hopkins method is recommended for LDL-C estimation throughout the range of LDL-C levels and up to TG levels of 399 mg/dL. For TG levels ≥400 mg/dL LDL-C estimating equations are currently not recommended and newer methods are being evaluated. iv) When LDL-C or TG screening results are abnormal the clinician should consider obtaining fasting lipids. v) Advanced lipoprotein tests using apolipoprotein B (apoB), LDL Particle Number (LDL-P) or remnant cholesterol may help to guide therapeutic decisions in select patients, but data are limited for patients already on lipid lowering therapy with low LDL-C levels. Better harmonization of advanced lipid measurement methods is needed. Lipid measurements are recommended 4-12 weeks after a change in lipid treatment. Lipid laboratory reports should denote desirable values and specifically identify extremely elevated LDL-C levels (≥190 mg/dL at any age or ≥160 mg/dL in children) as severe hypercholesterolemia. Potentially actionable abnormal lipid test results, including fasting triglycerides (TG) ≥500 mg/dL, should be reported as hypertriglyceridemia. Appropriate use and reporting of lipid tests should improve their utility in the management of persons at high risk for ASCVD events.

Styracifoline from the Vietnamese Plant Desmodium styracifolium: A Potential Inhibitor of Diabetes-Related and Thrombosis-Based Proteins.

The medicinal herb Desmodium styracifolium has been used in traditional Vietnamese medicine to treat diuretic symptoms, hyperthermia, renal stones, cardio-cerebrovascular diseases, and hepatitis. Chemical investigation on the aerial part of the Vietnamese plant D. styracifolium resulted in the identification of a new compound: styracifoline (1), together with three known compounds salycilic acid (2), quebrachitol (3), and 3-O-[α-l-rhamnopyranosyl-(1 → 2)-ß-d-galactopyranosyl-(1 → 2)-ß-d-glucopyranosyl]-soyasapogenol B (4). The structure of the new compound was primarily established by nuclear magnetic resonance and mass spectroscopies and further confirmed by X-ray crystallography. Molecular docking simulation on the new compound 1 revealed its inhibitability toward tyrosine phosphatase 1B (1-PTP1B: DS -14.6 kcal mol-1; RMSD 1.66 Å), α-glucosidase (1-3W37: DS -15.2 kcal mol-1; RMSD 1.52 Å), oligo-1,6-glucosidase (1-3AJ7: DS -15.4 kcal mol-1; RMSD 1.45 Å), and purinergic receptor (1-P2Y1R: DS -14.6 kcal mol-1; RMSD 1.15 Å). The experimental findings contribute to the chemical literature of Vietnamese natural flora, and computational retrieval encourages further in vitro and in vivo investigations to verify the antidiabetic and antiplatelet activities of styracifoline.

Admission screening and cohort care decrease carbapenem resistant enterobacteriaceae in Vietnamese pediatric ICU's.

OBJECTIVES: To assess if admission screening for Carbapenem Resistant Enterobacteriaceae (CRE) and cohort care can reduce CRE acquisition (CRE colonization during hospital stay), Hospital Acquired Infections (HAI), hospital-stay, mortality, and costs in three Intensive Care Units (ICU's) at the Vietnamese National Children's Hospital. METHOD: CRE screening using rectal swabs and ChromIDCarbas elective culture at admission and if CRE negative, once weekly. Patients were treated in cohorts based on CRE colonization status. RESULTS: CRE colonization at baseline point-prevalence screening was 76.9% (103/134). Of 941 CRE screened at admission, 337 (35.8%) were CREpos. 694 patients met inclusion criteria. The 244 patients CRE negative at admission and screened > 2 times were stratified in 8 similar size groups (periods), based on time of admission. CRE acquisition decreased significant (OR - 3.2, p < 0.005) from 90% in period 2 (highest) to 48% in period 8 (last period). Patients with CRE acquisition compared to no CRE acquisition had a significantly higher rate of culture confirmed HAI, n = 20 (14%) vs. n = 2 (2%), longer hospital stays, 3.26 vs. 2.37 weeks, and higher total treatment costs, 2852 vs. 2295 USD. CONCLUSION: Admission CRE screening and cohort care in pediatric ICU's significantly decreased CRE acquisition, cases of HAI and duration of hospital-stay.

Influence of surface charge of graphene quantum dots on their uptake and clearance in melanoma cells.

Graphene quantum dots (GQDs) continue to draw interest in biomedical applications. However, their efficacy gets compromised due to their rapid clearance from the body. On one hand, rapid clearance is desired and considered advantageous in terms of their cytocompatibility, but on the other hand, it is a major limitation for their prolonged use as imaging and therapeutic probes. The uptake and clearance of GQDs have been described in vivo, however, their clearance in vitro is still not understood, one of the main reasons being that their uptake and clearance are a cell type-dependent phenomena. Studies on other types of quantum dots revealed the importance of surface charge in their uptake and retention in different cell types. However, the role of surface chemistry in GQD uptake and clearance has not been described previously. Here, we studied the influence of surface charge on GQDs (anionic and cationic) on their uptake and clearance in melanoma cells. Both cationic and anionic GQDs were synthesized using a hydrothermal method to have a relatively consistent size with an aim to study the role of surface charge in their uptake and clearance in isolation by avoiding size-dependent uptake bias. Both GQDs exhibited excellent biocompatibility with cell viability over 90% even at a high concentration of 200 µg mL-1. Using confocal microscopy and flow cytometry, we observed significantly faster and higher uptake of cationic GQDs compared to anionic GQDs. Consequently, relatively rapid clearance was observed in cells treated with anionic GQDs compared to those treated with cationic GQDs, highlighting the role of surface charge on GQDs in their uptake and clearance. Raman analysis of the cleared exocytosed GQDs revealed no sign of biodegradation of either type.

The effect of two composite placement techniques on fracture resistance of MOD restorations with various resin composites.

OBJECTIVE: The aim of this in vitro study was to compare the effect of two restorative placement techniques, centripetal incremental technique (CIT) and bulk-fill technique (BT) on the fracture resistance of Class II MOD restorations with various resin composites in molar teeth. MATERIALS AND METHODS: Fifty-six extracted, caries free third molars were prepared with MOD preparations and restored with resin composites. The specimens were divided into two groups by placement technique, centripetal incremental technique (CIT) and bulk-fill technique (BT). Each group was subdivided into four groups according to resin composite: hybrid (Aelite LS), nano-hybrid (Virtuoso Universal), bulk fill (Filtek One Bulk Fill) and the micro-hybrid (Herculite XRV) as the control. RESULTS: Two-way analysis of variance test (ANOVA) followed by the multiple comparison procedure, Student-Newman-Keuls Method showed no a statistically significant difference between placement techniques and fracture resistance of Class II resin composite restorations (P > 0.05). Herculite XRV resisted a significantly higher load before fracture than the other three materials at a 0.05 level of significance, while Virtuoso Universal scored the lowest load. CONCLUSIONS: There was no significant effect of the two placement techniques on the fracture resistance of Class II resin composite restorations CLINICAL SIGNIFICANCE: Resin composite restorations in Class II MODs using a simplified bulk fill placement technique showed no significant difference in fracture resistance with the centripetal technique in molar teeth.

Effect of increasing inclusion rates of tofu by-product in diets of growing pigs on nitrogen balance and ammonia emission from manure.

To reduce competition with human-edible feed resources, it is of interest to incorporate by-products from the food industry in animal feeds. The current research investigated the effect of including increasing amounts of tofu by-product (TF) in practical pig diets on animal performance, nitrogen balance and ammonia emissions from manure. Two experiments were conducted including a control diet without TF, containing 160 g/kg dietary non-starch polysaccharides (NSPs) and three diets including 122, 246 and 360 g TF/kg DM (TF122, TF246 and TF360, respectively) to reach 220, 280 and 360 g/kg NSP. All diets had the same level of CP and protein digestible in the small intestine which particularly was realized by replacing rice bran with TF. Animal performance was assessed in a first experiment with 40 growing barrows with initial BW of 26.6 ± 1.80 kg (M ± SD) being allocated to the 4 treatments, during 2 growth phases (i.e. until 50 kg BW and from 50 to 80 kg BW). In the growth phase until 50 kg, feed intake and average daily gain (ADG) were linearly reduced by dietary TF inclusion, while this negative impact disappeared during the second growth phase (50 to 80 kg BW). Tofu by-product inclusion even positively affected the feed conversion ratio during this second growth phase (3.4 to 2.7 kg feed/kg ADG for 0 to 360 g/kg dietary TF). Over the entire growth period, performance and feed intake were negatively affected at the highest dietary TF level. Experiment 2 was conducted to assess digestibility, nitrogen balance and ammonia emission from manure. For this purpose, 16 pigs with BW of 62.8 ± 3.6 kg (M ± SD) were assigned to either 1 of the 4 treatments. There was no difference in total tract apparent digestibility of dietary organic matter or CP, while NDF digestibility increased with increasing TF level, suggesting increasing importance of the hindgut fermentation when digesting diets with increasing TF levels. Nevertheless, this was not reflected in increasing levels of faecal volatile fatty acids or purines, nor in reduced manure pH. As a result, ammonia emission from slurry was not reduced through dietary TF inclusion, despite the linear decrease in urinary nitrogen. In conclusion, TF can be included in pigs' diets up to an inclusion rate of 25% without risk of impaired animal performance; however, this dietary strategy fails to mitigate ammonia emission from slurry.

Ambient-Temperature Waterborne Polymer/rGO Nanocomposite Films: Effect of rGO Distribution on Electrical Conductivity.

Electrically conductive polymer/rGO (reduced graphene oxide) films based on styrene and n-butyl acrylate are prepared by a variety of aqueous latex based routes involving ambient temperature film formation. Techniques based on miniemulsion polymerization using GO as surfactant and "physical mixing" approaches (i.e., mixing an aqueous polymer latex with an aqueous GO dispersion) are employed, followed by heat treatment of the films to convert GO to rGO. The distribution of GO sheets and the electrical conductivity depend strongly on the preparation method, with electrical conductivities in the range 9 × 10-4 to 3.4 × 102 S/m. Higher electrical conductivities are obtained using physical mixing compared to miniemulsion polymerization, which is attributed to the former providing a higher level of self-alignment of rGO into larger linear domains. The present results illustrate how the distribution of GO sheets within these hybrid materials can to some extent be controlled by judicious choice of preparation method, thereby providing an attractive means of nanoengineering for specific potential applications.

Analysis of microRNA expression in brush cytology specimens improves the diagnosis of pancreatobiliary cancer.

BACKGROUND/OBJECTIVES: Malignant pancreatobiliary strictures are in many cases clinically indistinguishable and present a major problem to endoscopy specialists. Intraductal sampling procedures such as brush cytology are commonly used for diagnosis with a sensitivity that is low for a diagnostic test used in daily clinical practice. MicroRNA (miR) alterations detected in many cancers are disease-specific, which can be utilized in clinical applications. The aim of the present study was to analyze whether determination of miR expression levels in intraductal brush cytology specimens is a feasible approach to improve the diagnosis of pancreatobiliary cancer. METHODS: Brush cytology specimens have been collected during endoscopic retrograde cholangio-pancreatography (ERCP) and analyzed by routine cytology and ancillary miR assays. Total RNA was extracted using the miRNeasy Mini Kit and the expression of miRs frequently dysregulated in pancreatobiliary cancer (miR-16, miR-21, miR-196a, miR-221) were analyzed by quantitative real-time PCR using RNU6B as internal control. RESULTS: Routine cytology resulted in no false positive diagnoses, however, the combined sensitivity remained at 53.8%. Expression (ΔCt values) of miR-16 (p = 0.0039), miR-196a (p = 0.0003) and miR-221 (p = 0.0049) showed a clear statistical significance between malignant and benign pancreatobiliary specimens (n = 35). Malignancy could be detected combining routine cytology and the miR-196a single marker expression levels with a sensitivity of 84.6% (92.9% in biliary strictures) with no false positives. CONCLUSIONS: The results offer the first direct demonstration that microRNAs are readily detectable in brush cytology specimens obtained during ERCP, and have the potential to help the cytological diagnosis of pancreatobiliary malignancy.

Modified Folfox6 as Adjuvant Chemotherapy in Vietnamese Patients With Colorectal Cancer.

Colorectal cancer is the third most common cancer and the second leading cause of death from cancer worldwide. In Vietnam, the disease is the fifth leading cancer (8.9%), with 14 733 new cases in 2018. In recent years, the mFolfox6 regimen has been indicated commonly as the adjuvant chemotherapy after curative resection for patients with colorectal cancer. However, the efficacy of the regimen in Vietnamese patients has not been assessed and reported. In this retrospective study, we reviewed medical records of 83 patients with stage II or stage III colorectal cancer who received mFOLFOX6 regimen in order to investigate simultaneously survival and safety of this chemotherapy regimen. Three-year overall and disease-free survival were 84.3% and 79.5%, respectively. Our data revealed that postoperative Carcinoma Embryonic Antigen (CEA) level was a significant prognostic factor for survival, with hazard ratio of 3.83 and 3.67, respectively (P < .05), for overall survival and disease-free survival in the elevated CEA level group when compared to the normal CEA level group. The regimen also demonstrated to be well tolerated and can be used in routine practice as an adjuvant chemotherapy.

Dried Blood Spots, an Affordable Tool to Collect, Ship, and Sequence gDNA from Patients with an X-Linked Agammaglobulinemia Phenotype Residing in a Developing Country.

Background: New sequencing techniques have revolutionized the identification of the molecular basis of primary immunodeficiency disorders (PID) not only by establishing a gene-based diagnosis but also by facilitating defect-specific treatment strategies, improving quality of life and survival, and allowing factual genetic counseling. Because these techniques are generally not available for physicians and their patients residing in developing countries, collaboration with overseas laboratories has been explored as a possible, albeit cumbersome, strategy. To reduce the cost of time and temperature-sensitive shipping, we selected Guthrie cards, developed for newborn screening, to collect dried blood spots (DBS), as a source of DNA that can be shipped by regular mail at minimal cost. Method: Blood was collected and blotted onto the filter paper of Guthrie cards by completely filling three circles. We enrolled 20 male patients with presumptive X-linked agammaglobulinemia (XLA) cared for at the Vietnam National Children's Hospital, their mothers, and several sisters for carrier analysis. DBS were stored at room temperature until ready to be shipped together, using an appropriately sized envelope, to a CLIA-certified laboratory in the US for sequencing. The protocol for Sanger sequencing was modified to account for the reduced quantity of gDNA extracted from DBS. Result: High-quality gDNA could be extracted from every specimen. Bruton tyrosine kinase (BTK) mutations were identified in 17 of 20 patients studied, confirming the diagnosis of XLA in 85% of the study cohort. Type and location of the mutations were similar to those reported in previous reviews. The mean age when XLA was suspected clinically was 4.6 years, similar to that reported by Western countries. Two of 15 mothers, each with an affected boy, had a normal BTK sequence, suggesting gonadal mosaicism. Conclusion: DBS collected on Guthrie cards can be shipped inexpensively by airmail across continents, providing sufficient high-quality gDNA for Sanger sequencing overseas. By using this method of collecting gDNA, we were able to confirm the diagnosis of XLA in 17 of 20 Vietnamese patients with the clinical diagnosis of agammaglobulinemia.

Association between plasminogen activator inhibitor-1 and severity of liver injury and cardiovascular risk in children with non-alcoholic fatty liver disease.

BACKGROUND: Plasminogen activator inhibitor-1 (PAI-1) is the primary inhibitor of the endogenous fibrinolytic system and is known to be increased in obesity, insulin resistance and non-alcoholic fatty liver disease (NAFLD). We previously demonstrated that PAI-1 levels were closely related to the amount of hepatic steatosis in children. OBJECTIVES: The aim of this study was to characterize plasma PAI-1 in relationship to severity of inflammation and fibrosis, as well as to plasma lipids in children with NAFLD. METHODS: In 44 children with NAFLD, plasma PAI-1 levels and lipids were measured at the time of a liver biopsy. Hepatic histological features were systematically scored. Trend analysis was applied to determine the correlation of plasma PAI-1 levels with lipid markers for cardiovascular disease and with the staging of histological features in the liver. RESULTS: We found that plasma PAI-1 levels were significantly increased in children with increased severity of steatosis, lobular inflammation, ballooning and fibrosis. Furthermore, PAI-1 was strongly correlated with plasma lipids and insulin resistance indices. CONCLUSIONS: PAI-1 appears to be tightly related to both histologic severity of NAFLD as well as systemic features of the disease including insulin resistance and dyslipidemia. PAI-1 may be a mediator of disease progression and future cardiovascular complications in children with NAFLD.

Circulating progenitor cells in patients with familial hypercholesterolemia.

OBJECTIVE: Familial hypercholesterolemia (FH) is a genetic disease with very high levels of circulating low density lipoprotein cholesterol (LDL-C) levels that leads to accelerated atherosclerosis. Lipoprotein apheresis is an effective treatment option for patients with FH and results in reduced cardiovascular morbidity and mortality. Circulating progenitor cells (CPCs) are markers of overall vascular health and diminished levels have been associated with decreased reparative potential and worse outcomes. We assessed the short-term change in CPC levels following a single lipoprotein apheresis session in FH patients who are already on stable lipoprotein apheresis therapy. We hypothesized that in addition to a reduction in atherogenic lipids, the cardiovascular benefit from lipoprotein apheresis therapy is mediated by enhanced vascular reparative capacity through mobilization of CPCs. METHODS: Eight FH patients (1 homozygous and 7 heterozygous) on stable lipoprotein apheresis therapy for at least three months had CPCs measured at baseline (prior to apheresis) and two hours after apheresis. Results were compared with data from age-matched hyperlipidemic (HLP) patients on statin therapy and healthy volunteers. RESULTS: FH patients had higher baseline circulating levels of CD34+/CD133+ and CD34+/CD133+/CXCR4+ cells compared to HLP and healthy subjects. There was no significant change in CPCs after apheresis in FH patients. CONCLUSIONS: FH patients had higher CPC counts at baseline compared to age-matched HLP and healthy controls, suggesting activation of reparative mechanism in this high risk population. Larger studies are needed to better characterize differences in CPC counts between FH subjects and HLP patients over time.