Search

Background: This study aims to assess the long-term outcomes of a modified pneumatic reduction protocol for intussusception at the Vietnam National Hospital of Pediatrics, an institution with a significant patient load in a lower-middle-income country., Patients and Methods: A single center, retrospective cohort observational study was conducted to examine patients who underwent modified fluoroscopic-guided air-enema reduction (FGAR) for intussusception from January 2016 to December 2017. Data on patient demographics, complication rates, and the incidence of long-term recurrence was collected., Results: Between January 2016 and December 2017, a total of 3562 patients underwent modified FGAR at our institution, including 2313 males (64.9%) and 1249 females (35.1%). The median age was 19 months (range: 1-170), and the median FGAR procedure duration was 4 min (range: 2-24). The median hospital stay was 1 day (range: 1-31). Successful reduction was achieved in 98.7% of cases, with 43 unsuccessful cases and 4 cases of perforated bowel requiring surgery. Twenty patients, presenting with severe symptoms due to delayed treatment seeking, were admitted to the pediatric intensive care unit (ICU) post-FGAR. No mortality or severe morbidity was reported. Over a median 6-year follow-up, intussusception recurred in 198 patients, accounting for 5.6% of the cohort, with 97% of recurrences occurring within the first year post-reduction. Infants and children under 12 months of age had the highest complication rates, including failed FGAR, complicated intussusception, ICU admission, or recurrence, compared to other age groups, and this difference was statistically significant (p < 0.05)., Conclusion: The modified FGAR protocol has been demonstrated to be safe and feasible, with a very high success rate, low complication rate, and low recurrence rate. Although further comparative studies are needed to confirm its reproducibility, it should be considered a promising approach for children in low-to middle-income countries., Level of Evidence: Level III., Competing Interests: Conflict of interest None., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Recently, researchers have uncovered a correlation between loneliness and both the development and management of diabetes. Nevertheless, previous studies employing an unvalidated loneliness questionnaire impair result accuracy. Furthermore, this aspect has not been researched in the Japanese population. Therefore, this cross-sectional study analyzed data from the Kanagawa prospective "ME-BYO" Cohort Study (ME-BYO cohort) to investigate the correlation between loneliness, as measured by 20 items on the UCLA Loneliness Scale, and blood glucose levels. A total of 666 participants were included in the analysis, with a mean age of 54.1 years and a mean BMI of 23 kg/m 2 . Half of the participants had obtained an education level beyond high school. The mean household income and physical activity level were reported as 6.83 million Japanese yen and 12.3 METs-h/day, respectively. Model 1 of the linear regression analysis determined that there was no significant association between the loneliness scale and HbA1c (p = 0.512). After adjusting for age, gender, BMI (model 2), sitting time, physical activity level (model 3), housemates, household income, and final education (model 4), and controlling for social support, quality of life, and depression (model 5), the results showed no significant association, with a p-value of 0.823, 0.791, 0.792, and 0.816, respectively. Thus, the study found no link between loneliness and HbA1c in the high SES population. This finding contradicts previous results and may be attributed to the impact of population characteristics, SES status, or genetic backgrounds., (© 2024. The Author(s).)

Background: The short-term association between increasing temperatures and injury has been described in high-income countries, but less is known for low-income and-middle-income countries, including Vietnam., Methods: We used emergency injury visits (EIV) data for 2017-2019 from 733 hospitals and clinics in Hanoi, Vietnam to examine the effects of daily temperature on EIV. Time-series analysis with quasi-Poisson models was used to estimate a linear relative risk increase (RRI) for overall populations and ones stratified by age and sex. Exposure-response curves estimated non-linear associations as an RR between daily temperature and injury. Models were adjusted for the day of week, holidays, daily relative humidity, daily particulate matter, and long-term and seasonal trends., Results and Conclusions: A total of 39 313 EIV were recorded averaging 36 injuries daily. Injuries more likely occurred in males and those aged 15-44, and aged 44-60. For linear effects, a 5°C increase in same day mean temperature was associated with an overall increased EIV (RRI 4.8; 95% CI 2.3 to 7.3) with males (RRI 5.9; 95% CI 3.0 to 8.9) experiencing a greater effect than females (RRI 3.0; 95% CI -0.5 to 6.5). Non-linear effects showed an increase in EIV at higher temperatures compared with the threshold temperature of 15°C, with the greatest effect at 33°C (RR 1.3; 95% CI 1.2 to 1.6). Further research to investigate temperature-injury among different populations and by the cause of injury is warranted., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Research over the past four decades has built a convincing case that co-speech hand gestures play a powerful role in human cognition . However, this recent focus on the cognitive function of gesture has, to a large extent, overlooked its emotional role-a role that was once central to research on bodily expression. In the present review, we first give a brief summary of the wealth of research demonstrating the cognitive function of co-speech gestures in language acquisition, learning, and thinking. Building on this foundation, we revisit the emotional function of gesture across a wide range of communicative contexts, from clinical to artistic to educational, and spanning diverse fields, from cognitive neuroscience to linguistics to affective science. Bridging the cognitive and emotional functions of gesture highlights promising avenues of research that have varied practical and theoretical implications for human-machine interactions, therapeutic interventions, language evolution, embodied cognition, and more., (© 2023 The Authors. Topics in Cognitive Science published by Wiley Periodicals LLC on behalf of Cognitive Science Society.)

The effects of temperature on behavior change and mental health have previously been explored, but the association between temperature and crime is less well understood, especially in developing countries. Single-city-level data were used to evaluate the association between the short-term effects of temperature on crime events in urban Hanoi, Vietnam. We used quasi-Poisson regression models to investigate the linear effects and distributed lag non-linear models to investigate the non-linear association between daily temperature and daily crime events from 2013 to 2019. There were 3884 crime events, including 1083 violent crimes and 2801 non-violent crimes during the 7-year study period. For both linear and non-linear effects, there were positive associations between an increase in daily temperature and crime, and the greatest effects were observed on the first day of exposure (lag 0). For linear effects, we estimated that each 5 °C increase in daily mean temperature was associated with a 9.9% (95%CI: 0.2; 20.5), 6.8% (95%CI: 0.6; 13.5), and 7.5% (95%CI: 2.3; 13.2) increase in the risk of violent, non-violent, and total crime, respectively. For non-linear effects, however, the crime risk plateaued at 30 °C and decreased at higher exposures, which presented an inverted U-shape response with a large statistical uncertainty.

This study aimed to examine the short-term effects of ambient temperature on hospital admissions due to respiratory diseases among Hanoi residents. We collected 34,653 hospital admissions for 365 days (November 1, 2017, to November 31, 2018) from two hospitals in Hanoi. A quasi-Poisson regression model with time series analysis was used to explore the temperature-health outcome relationship's overall pattern. The non-linear curve indicated the temperatures with the lowest risk range from 22 degrees (Celcius) to 25 degrees (Celcius). On average, cold temperatures showed a higher risk than hot temperatures across all genders and age groups. Hospital admissions risk was highest at 13 degrees (Celcius) ( RR = 1.39; 95% CI = 1.26-1.54) for cold effects and at 33 degrees (Celcius) ( RR = 1.21, 95% CI = 1.04-1.39) for the hot effects. Temporal pattern analysis showed that the most effect on respiratory diseases occurred at a lag of 0 days for hot effect and at a lag of 1 day for cold effect. The risk of changing temperature among women and people over 5 years old was higher than other groups. Our results suggest that the risk of respiratory admissions was greatest when the temperature was low. Public health prevention programs should be enhanced to improve public awareness about the health risks of temperature changes, especially respiratory diseases risked by low temperatures., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Tran, Le, Ngo, Le, Phung, Berman and Nguyen.)

Sirtuin-3 (Sirt3) is a major mitochondrial deacetylase enzyme that regulates multiple metabolic pathways, and its expression is decreased in diabetes type 1 and type 2 diabetes. This study aimed to elucidate Sirt3's molecular mechanism in regulating insulin sensitivity in adipocytes that can contribute to the effort of targeting Sirt3 for the treatment of obesity and type 2 diabetes. We found that the Sirt3 activator honokiol (HNK) induced adipogenesis compared to the control, in contrast to Sirt3 inhibitor, 3-TYP. Accordingly, HNK increased expression of adipocyte gene markers, gene-involved lipolysis and glucose transport (GLUT4), while 3-TYP reduced expression of those genes. Interestingly, 3-TYP caused an increase in gene expression of adipocyte-specific cytokines including IL6, resistin, and TNF-α. However, changes in adipocyte-specific cytokines in HNK treated cells were not significant. In addition, HNK stimulated insulin pathway by promoting insulin receptor beta (IRβ) and PI3K/AKT/mTOR pathways, resulting in an increase in phosphorylation of the forkhead family FoxO1/FoxO3a/FoxO4 and glycogen synthase kinase-3 (GSK-3β), opposing 3-TYP. In line with these findings, HNK increased free fatty acid and glucose uptake, contrary to 3-TYP. In conclusion, Sirt3 activator-HNK induced adipogenesis and lipolysis reduced adipocytes specific cytokines. Intriguingly, HNK activated insulin signaling pathway and increased free fatty acid as well as glucose uptake and transport, in sharp contrast to 3-TYP. These results indicate that, via insulin signaling regulation, Sirt3 activation by HNK improves insulin resistance, while Sirt3 inhibition by 3-TYP might precipitate insulin resistance.

Biliary atresia (BA) is a progressive inflammation and fibrosis of the biliary tree characterized by the obstruction of bile flow, which results in liver failure, scarring and cirrhosis. This study aimed to explore the elusive aetiology of BA by conducting whole exome sequencing for 41 children with BA and their parents (35 trios, including 1 family with 2 BA-diagnosed children and 5 child-mother cases). We exclusively identified and validated a total of 28 variants (17 X-linked, 6 de novo and 5 homozygous) in 25 candidate genes from our BA cohort. These variants were among the 10% most deleterious and had a low minor allele frequency against the employed databases: Kinh Vietnamese (KHV), GnomAD and 1000 Genome Project. Interestingly, AMER1, INVS and OCRL variants were found in unrelated probands and were first reported in a BA cohort. Liver specimens and blood samples showed identical variants, suggesting that somatic variants were unlikely to occur during morphogenesis. Consistent with earlier attempts, this study implicated genetic heterogeneity and non-Mendelian inheritance of BA., (© 2021. The Author(s).)

The California Bearing Ratio (CBR) is an important index for evaluating the bearing capacity of pavement subgrade materials. In this research, random subspace optimization-based hybrid computing models were trained and developed for the prediction of the CBR of soil. Three models were developed, namely reduced error pruning trees (REPTs), random subsurface-based REPT (RSS-REPT), and RSS-based extra tree (RSS-ET). An experimental database was compiled from a total of 214 soil samples, which were classified according to AASHTO M 145, and included 26 samples of A-2-6 (clayey gravel and sand soil), 3 samples of A-4 (silty soil), 89 samples of A-6 (clayey soil), and 96 samples of A-7-6 (clayey soil). All CBR tests were performed in soaked conditions. The input parameters of the models included the particle size distribution, gravel content (G), coarse sand content (CS), fine sand content (FS), silt clay content (SC), organic content (O), liquid limit (LL), plastic limit (PL), plasticity index (PI), optimum moisture content (OMC), and maximum dry density (MDD). The accuracy of the developed models was assessed using numerous performance indexes, such as the coefficient of determination, relative error, MAE, and RMSE. The results show that the highest prediction accuracy was obtained using the RSS-based extra tree optimization technique.

Introduction: Medical students have been serving as a key part of the frontline health workforce responding to the coronavirus disease 2019 (COVID-19) pandemic globally. Their contribution is especially important in the resource-scarce settings of developing nations such as Vietnam. Yet, the intention of medical students, in particular, nursing students, to participate in COVID-19 frontline prevention activities has not been well-understood. This study aimed to examine factors associated with the intentionto participate in COVID-19 frontline prevention activities among Vietnamese nursing students. Methods: A cross-sectional study was conducted on a total of 597 students in December 2020 in Hanoi, Vietnam. Information regarding the socioeconomic characteristics of participants, their source of COVID-19 related knowledge, and their perception and attitude toward participating in COVID-19 frontline activities [based on Theory of Planned Behavior (TPB)] was collected. A hierarchical regression model was employed to examine the association between intentions of students and associated factors. Results: A positive intention to participate in COVID-19 frontline prevention activities was found (mean score of 25.3 over 35; SD = 4.4; min = 5; max = 35). Attitude toward behavior, subjective norms, and perceived behavioral control (PBC) was found to be significantly associated with the intention of students. These variables explained the 37% variation in the intention of students in the model. Among three factors, subjective norm showed the strongest correlation with intention of students (β = 0.358; p < 0.001). Obtaining information from official sources and community was also found to be positively correlated with intention to participate. Conclusion: Most of the respondents reported a positive intention to participate in COVID-19 frontline prevention activities. The findings suggested that the TPB was a good instrument to predict the intention to perform behavior among Vietnamese students. Enhancing the positive attitude of students, encouraging family and community supports, and providing adequately essential resources will contribute to optimizing the participation of students to confront COVID-19., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Tran, Nguyen, Bui, Tran, Nguyen, Nguyen, Nguyen and Nguyen.)

Background: To present a surgical technique of single-incision laparoscopic-assisted endorectal pull-through (SILEP) with suspension sutures using conventional instruments for Hirschsprung disease (HD) and its long-term follow-up outcomes., Methods: The procedure began with a 1 cm transumbilical skin incision. Three separate punctures were made in the fascia with a 5 mm scope in the middle and 5 mm and 3 mm ports for working instruments on the left and right, respectively. The first suspension suture was placed to secure the sigmoid colon to the abdominal wall. A window was created through the rectal mesentery, and dissection around the rectum was carried out. The second suspension suture was performed to suspend the rectovesical peritoneal fold or the rectovaginal peritoneal fold to the abdominal wall. Dissection around the rectum was continued downward to approximately 1 cm below the peritoneal fold. Then, the operation was completed by a transanal approach., Results: Forty patients underwent SILEP from March 2013 to April 2015. The median age was 2.7 months (ranging from 1 to 17 months). The mean operative time was 96 ± 23 min. No conversion to an open operation was required. The average hospitalization time was 4.5 ± 2 days. There were no intraoperative or perioperative complications. Long-term follow-up results were obtained from 38 patients. A frequency of defecation from every other day to twice a day was noted for 33 patients (86.8%) and more often for 5 patients (13.2%). Two patients had enterocolitis (5.2%)., Conclusion: Single-incision laparoscopic rectal pull-through with suspension sutures using conventional instruments is feasible and safe for HD with good long-term outcomes.

We conducted a cross-sectional study in four provinces located in four different geographical areas in Vietnam to examine the prevalence and associated factors of depression and suicide ideation among school students aged 13-17. A sample of 6407 students from secondary school and high school participated in our survey to complete self-reported questionnaires. Depressive symptoms were measured by the Center for Epidemiologic Studies Depression scale (CES-D). Suicidal ideation and associated factors were measured by the Global School Student Health Survey (GSHS) questionnaire. We found that 31.7% of students had depressive symptoms, and 11% reported suicidal ideation during the last year. Female students and older students were more at risk of experiencing depressive symptoms and suicidal ideation than male students and younger students. Bullying, violence, smoking, and alcohol consumption appear as risk factors, while a good relationship with parents/guardians may protect school students from having depressive symptoms and suicidal ideation., Competing Interests: Declaration of conflicting interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2020.)

The spread of avian influenza virus among Asian countries is becoming a concern after influenza epidemics in recent years. This study is aimed at identifying the subtypes of avian influenza viruses collected from healthy chickens and ducks at two live bird markets in a border province of Vietnam and the Lao People Democratic Republic. Cloacal and tracheal swab samples from 100 chickens and 101 ducks were collected in May 2017. All samples were screened to detect avian influenza virus by real-time reverse transcriptase PCR. Samples that are avian influenza virus-positive were isolated in embryonated chicken eggs, and the subtypes were identified by RT-PCR with the specific primers. The samples positive for influenza virus H5 were sequenced to identify HA and NA genes. The prevalence of avian influenza virus (AIV) among chicken and duck samples was 27.5% (55/200) and 24.8% (50/202), respectively. AIV subtypes identified among 17 samples positive with the hemagglutination test include H3N6, H6N6, and H9N2. Of these 17 samples, 7 duck samples were found to be H6N6, 4 duck samples were infected with both subtypes of H3N6 and H6N6, and two chicken samples were recorded as H9N2. A positive chicken sample with A/H5 contains 99% similarity nucleotide with H5N6 reference strain. Results suggested that while the presence of low pathogenic avian influenza virus is predominant, potential risks of the appearance of high pathogen avian influenza virus in the east-west boundary in Vietnam should be concerned and studied further. Furthermore, prevention activities are needed to reduce such biosecurity threats in Vietnam and other Asian countries., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2020 Quynh Anh Tran et al.)

This study assessed the diagnostic value of interleukin- (IL-) 6, high-sensitivity C-reactive protein (hs-CRP), and procalcitonin (PCT) in differentiating severe pneumonia caused by respiratory syncytial virus (RSV) alone and RSV with bacterial coinfections among Vietnamese children under 5 years old. A cross-sectional study on 70 children with severe RSV pneumonia was conducted. IL-6, hs-CRP, and PCT tests were performed. Receiver operating characteristic (ROC) analysis was employed to measure the diagnostic values of PCT, IL-6, and hs-CRP. Of 70 children, 11 children were confirmed to have bacterial coinfections. The most common bacterial coinfection was Haemophilus influenzae . This study underlined that inflammatory biomarkers such as PCT had a moderate-to-high capability of disseminating severe pneumonia children with RSV alone or RSV and bacterial coinfections. This may support clinicians in administrating appropriate antibiotics to children suffering from severe RSV pneumonia., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2020 Quyet Do et al.)

Background: Laparoscopic-assisted endorectal pull-through (LAEPT) procedure in the management of high-type anorectal malformations (ARMs) was first introduced in 1998 and is quickly accepted worldwide. However, evidence on long-term outcomes of this technique is constrained. This study aims to evaluate the long-term outcomes of LAEPT for high-type ARMs in Vietnamese pediatrics. Materials and Methods: A longitudinal study was carried out from January 2009 to July 2014 in 56 patients <3 years old. Variables included age of operation, associated anomalies, type of fistula, the duration of hospital stay, complications, and long-term functional outcomes (Krickenbeck modified standards were used for children ≥3 years). Results: There were 56 patients including 48 males and 8 females. The mean age at operation was 3.7 months, the mean hospital stay was 4.6 days. The mean operative time was 76.7 minutes. The mean follow-up time was from 38 to 104 months (mean follow-up: 71.5 months). There were 46 (82.1%) patients having feeling of urge, 42 (75.0%) patients having capacity to verbalize, and 40 (71.4%) patients having hold the bowel movement. Conclusion: LAEPT is feasible, safe, and effective in the management of high-type ARMs.

Developmental studies and 3D in vitro model systems show that the production and engagement of extracellular matrix (ECM) often precede stem cell differentiation. Yet, unclear is how the ECM triggers signaling events in sequence to accommodate multistep process characteristic of differentiation. Here, we employ transcriptome profiling and advanced imaging to delineate the specificity of ECM engagement to particular differentiation pathways and to determine whether specificity in this context is a function of long-term ECM remodeling. To this end, human mesenchymal stem cells (hMSCs) were cultured in 3D bioprinted prisms created from ECM proteins and associated controls. We found that exogenous ECM provided in 3D microenvironments at early time points impacts on the composition of microenvironments at later time points and that each evolving 3D microenvironment is uniquely poised to promote stem cell differentiation. Moreover, 2D cultures undergo minimal ECM remodeling and are ill-equipped to stimulate pathways associated with development.

Background: Young children with suspected abusive head trauma often receive skull radiographs to evaluate for fractures as well as computed tomography (CT) of the head to assess for intracranial injury. Using a CT as the primary modality to evaluate both fracture and intracranial injury could reduce exposure to radiation without sacrificing performance., Objective: To evaluate the sensitivity of CT head with (3-D) reconstruction compared to skull radiographs to identify skull fractures in children with suspected abusive head trauma., Materials and Methods: This was a retrospective (2013-2014) cross-sectional study of infants evaluated for abusive head trauma via both skull radiographs and CT with 3-D reconstruction. The reference standard was skull radiography. All studies were read by pediatric radiologists and neuroradiologists, with ten percent read by a second radiologist to evaluate for interobserver reliability., Results: One hundred seventy-seven children (47% female; mean/median age: 5 months) were included. Sixty-two (35%) had skull fractures by radiography. CT with 3-D reconstruction was 97% sensitive (95% confidence interval [CI]: 89-100%) and 94% specific (CI: 87-97%) for skull fracture. There was no significant difference between plain radiographs and 3-D CT scan results (P-value = 0.18). Kappa was 1 (P-value <0.001) between radiologist readings of CTs and 0.77 (P = 0.001) for skull radiographs., Conclusion: CT with 3-D reconstruction is equivalent to skull radiographs in identifying skull fractures. When a head CT is indicated, skull radiographs add little diagnostic value.

Recent systematic reviews have emphasized the need for more research into the health and social impacts of adverse childhood experiences (ACEs) in the Asia-Pacific region. This cross-sectional study was conducted with 2099 young adult students in 8 medical universities throughout Vietnam. An anonymous, self-report questionnaire included the World Health Organization ACE-International Questionnaire and standardized measures of mental and physical health. Three quarters (76%) of the students reported at least one exposure to ACEs; 21% had 4 or more ACEs. The most commonly reported adversities were emotional abuse, physical abuse, and witnessing a household member being treated violently (42.3%, 39.9%, and 34.6%, respectively). Co-occurrence of ACEs had dose-response relationships with poor mental health, suicidal ideation, and low physical health-related quality of life. This first multisite study of ACEs among Vietnamese university students provided evidence that childhood adversity is common and is significantly linked with impaired health and well-being into the early adult years., (© 2015 APJPH.)

The extracellular matrix (ECM) of the embryonic heart guides assembly and maturation of cardiac cell types and, thus, may serve as a useful template, or blueprint, for fabrication of scaffolds for cardiac tissue engineering. Surprisingly, characterization of the ECM with cardiac development is scattered and fails to comprehensively reflect the spatiotemporal dynamics making it difficult to apply to tissue engineering efforts. The objective of this work was to define a blueprint of the spatiotemporal organization, localization, and relative amount of the four essential ECM proteins, collagen types I and IV (COLI, COLIV), elastin (ELN), and fibronectin (FN) in the left ventricle of the murine heart at embryonic stages E12.5, E14.5, and E16.5 and 2 days postnatal (P2). Second harmonic generation (SHG) imaging identified fibrillar collagens at E14.5, with an increasing density over time. Subsequently, immunohistochemistry (IHC) was used to compare the spatial distribution, organization, and relative amounts of each ECM protein. COLIV was found throughout the developing heart, progressing in amount and organization from E12.5 to P2. The amount of COLI was greatest at E12.5 particularly within the epicardium. For all stages, FN was present in the epicardium, with highest levels at E12.5 and present in the myocardium and the endocardium at relatively constant levels at all time points. ELN remained relatively constant in appearance and amount throughout the developmental stages except for a transient increase at E16.5. Expression of ECM mRNA was determined using quantitative polymerase chain reaction and allowed for comparison of amounts of ECM molecules at each time point. Generally, COLI and COLIII mRNA expression levels were comparatively high, while COLIV, laminin, and FN were expressed at intermediate levels throughout the time period studied. Interestingly, levels of ELN mRNA were relatively low at early time points (E12.5), but increased significantly by P2. Thus, we identified changes in the spatial and temporal localization of the primary ECM of the developing ventricle. This characterization can serve as a blueprint for fabrication techniques, which we illustrate by using multiphoton excitation photochemistry to create a synthetic scaffold based on COLIV organization at P2. Similarly, fabricated scaffolds generated using ECM components, could be utilized for ventricular repair.

To understand complex micro/nanoscale ECM stem cell interactions, reproducible in vitro models are needed that can strictly recapitulate the relative content and spatial arrangement of native tissue. Additionally, whole ECM proteins are required to most accurately reflect native binding dynamics. To address this need, we use multiphoton excited photochemistry to create 3D whole protein constructs or "modules" to study how the ECM governs stem cell migration. The constructs were created from mixtures of BSA/laminin (LN) and BSA alone, whose comparison afforded studying how the migration dynamics are governed from the combination of morphological and ECM cues. We found that mesenchymal stem cells interacted for significantly longer durations with the BSA/LN constructs than pure BSA, pointing to the importance of binding cues of the LN. Critical to this work was the development of an automated system with feedback based on fluorescence imaging to provide quality control when synthesizing multiple identical constructs.

Although it is widely accepted that glucocorticoids (GC) are a mainstay of the treatment of diseases characterized by airway inflammation, little is known about the effects of GC on local innate immunity. In this article, we report that respiratory epithelial cells manifested a local "acute phase response" after stimulation with TLR activation and TNF-alpha and that GC spared or enhanced the epithelial expression of molecules that are involved in host defense, including complement, collectins, and other antimicrobial proteins. As expected, GC inhibited the expression of molecules responsible for inflammation such as cytokines (IFNbeta and GM-CSF) and chemokines (RANTES and IL-8). Studies using Western blotting, EMSA, and functional analysis indicated that the selective effects of GC are mediated through activation of the transcription factor C/EBP. Knockdown of C/EBPbeta by small interfering RNA blocked the enhancement by GC of host defense molecule expression but had no effect on inflammatory gene expression. These results suggest that GC spare or enhance local innate host defense responses in addition to exerting anti-inflammatory actions. It is possible that the known ability of GC to reduce the exacerbation of diseases in which infectious organisms serve as triggering factors (e.g., asthma, allergic bronchopulmonary aspergillosis, and chronic obstructive pulmonary disease) may result in part from enhanced innate immune responses in airway mucosa.

Background: The role of the innate immune system in the pathophysiology of chronic rhinosinusitis (CRS) is poorly understood. In this study, we compared sinonasal expression of toll-like receptors (TLRs), complement components, serum amyloid A, and inflammatory genes (chemokines and cytokines) in control subjects and patients undergoing sinus surgery for CRS., Methods: Eleven control subjects and 30 subjects with CRS unresponsive to medical management were enrolled prospectively before undergoing endoscopic sinus surgery. Ethmoid mucosal specimens were obtained surgically and processed for RNA extraction. Real-time polymerase chain reaction was used to quantitate the level of expression of messenger RNA (mRNA) for TLR, acute phase proteins, and cytokine genes. Subjects were followed for a minimum of 6 months postoperatively with nasal endoscopy to assess for recurrence of polyps., Results: mRNA for all target genes was detected in the ethmoid mucosa of both control and CRS subjects. The level of gene expression was normalized to the housekeeping genes 18s RNA and glyceraldehyde-3-phosphate dehydrogenase. As compared with controls, CRS was associated with significantly higher expression of TLR2 and the inflammatory genes macrophage-inflammatory protein alpha, RANTES, and granulocyte-macrophage colony-stimulating factor. Patients with early recurrence of polyps after surgery had significantly decreased expression of TLR2, 9, and serum amyloid A and increased expression of macrophage-inflammatory protein alpha compared with surgery-responsive patients., Conclusion: This study shows the increased levels of expression of TLR2 and a variety of inflammatory genes in sinonasal mucosa of CRS patients compared with controls. Whether these differences play a role in pathogenesis or are merely manifestations of disease activity is worthy of investigation.

Background: There is a growing appreciation of the role that nasal mucosa plays in innate immunity. In this study, the expression of pattern recognition receptors known as toll-like receptors (TLRs) and the effector molecules complement factor 3 (C3), properdin, and serum amyloid A (SAA) were examined in human sinonasal mucosa obtained from control subjects and patients with chronic rhinosinusitis (CRS)., Methods: Sinonasal mucosal specimens were obtained from 20 patients with CRS and 5 control subjects. Messenger RNA (mRNA) was isolated and tested using Taqman real-time polymerase chain reaction with primer and probe sets for C3, complement factor P, and SAA. Standard polymerase chain reaction was performed for the 10 known TLRs. Immunohistochemistry was performed on the microscopic sections using antibodies against C3., Results: Analysis of the sinonasal sample mRNA revealed expression of all 10 TLRs in both CRS samples and in control specimens. Expression of the three effector proteins was detected also, with the levels of mRNA for C3 generally greater than SAA and properdin in CRS patients. No significant differences were found in TLR or innate immune protein expression in normal controls. Immunohistochemical analysis of sinonasal mucosal specimens established C3 staining ranging from 20 to 85% of the epithelium present., Conclusion: These studies indicate that sinonasal mucosa expresses genes involved in innate immunity including the TLRs and proteins involved in complement activation. We hypothesize that local production of complement and acute phase proteins by airway epithelium on stimulation of innate immune receptors may play an important role in host defense in the airway and, potentially, in the pathogenesis of CRS.

Activated T cells have been implicated in chronic rhinosinusitis (CRS) and asthma and physically interact with epithelial cells in the airways. We now report that human airway epithelial cells display significant constitutive cell-surface expression of costimulatory ligands, B7-H1, B7-H2, B7-H3, and B7-DC. Expression of B7-H1 and B7-DC was selectively induced by stimulation of either BEAS2B or primary nasal epithelial cells (PNEC) with interferon (IFN)-gamma (100 ng/ml). The combination of IFN-gamma and tumor necrosis factor-alpha (100 ng/ml) selectively induced expression better than IFN-gamma alone. Fluticasone treatment (10(-7) M) reduced the baseline expression and inhibited the induction of B7-H1 and B7-DC in BEAS2B cells. In vitro exposure of PNEC to IFN-gamma also resulted in selective induction of B7-H1 and B7-DC. Monoclonal antibody blockade of B7-H1 or B7-DC enhanced IFN-gamma expression by purified T cells in co-culture experiments, suggesting that these two B7 homologs inhibit T cell responses at the mucosal surface. Immunohistochemical staining of human sinonasal surgical tissue confirmed the presence of B7-H1, B7-H2, and B7-H3 in the epithelial cell layer, especially in samples from patients diagnosed with Samter's Triad, a severe form of CRS. Real-time PCR analysis of sinonasal tissue revealed elevated levels of B7-H1 and B7-DC in CRS compared with controls. These results demonstrate that epithelial cells express functional B7 costimulatory molecules and that expression of selected B7 family members is inducible in vitro and in vivo. Epithelial B7 homologs could play a role in regulation of lymphocytic activity at mucosal surfaces.

The production and removal of regulatory RNAs must be controlled to ensure proper physiological responses. SsrA RNA (tmRNA), a regulatory RNA conserved in all bacteria, is cell cycle regulated and is important for control of cell cycle progression in Caulobacter crescentus. We report that RNase R, a highly conserved 3' to 5' exoribonuclease, is required for the selective degradation of SsrA RNA in stalked cells. Purified RNase R degrades SsrA RNA in vitro, and is kinetically competent to account for all SsrA RNA turnover. SmpB, a tmRNA-binding protein, protects SsrA RNA from RNase R degradation in vitro, and the levels of SmpB protein during the cell cycle correlate with SsrA RNA stability. These results suggest that SmpB binding controls the timing of SsrA RNA degradation by RNase R. We propose a model for the regulated degradation of SsrA RNA in which RNase R degrades SsrA RNA from a non-tRNA-like 3' end, and SmpB specifically protects SsrA RNA from RNase R. This model explains the regulation of SsrA RNA in other bacteria, and suggests that a highly conserved regulatory mechanism controls SsrA activity.