Pediatric proximal tibial fractures: How does the trauma mechanism guide the operative strategy?

PURPOSE: Pediatric proximal tibial fractures (PTF) are rare but potentially debilitating. So far, no system for guiding surgical treatment based on injury-force mechanism has been documented, while adult tibial plateau fractures have benefited greatly from such an approach. This study reviews the diagnosis and treatment experience at a tertiary trauma center and introduces the reduction-traction method. METHODS: Pediatric patients (0-17 years old) diagnosed with PTF were identified in the hospital database from 2017 to 2021. Their injury mechanism, injury location, treatment type, and treatment outcomes were recorded. Images were reviewed to establish an injury-force classification according to Mubarak et al., 2009. When appropriate, patients were treated using a "reduction-traction" approach. RESULTS: Twenty-nine patients were identified, and followed-up for a mean of 6.8 months. The most common cause of injury was falling from height < 2 m, often from a trampoline. The tibial plateau and proximal tibial metaphysis were most commonly involved. Thirteen patients were treated non-operatively, 10 with open reduction and internal fixation, and six with arthroscopic surgery. A bimodal distribution according to age was noted in the injury mechanism, injury site, and treatment type. No adverse outcomes were recorded, and all patients resumed sports activities. The "reduction-traction" technique produced favorable outcomes in three patients. CONCLUSIONS: Pediatric PTF has a bimodal distribution with high risk before three years and after 15 years. The injury-force classification can supplement the Salter-Harris classification in guiding surgical treatment. The "reduction-traction" approach in children differs from adults, and results in good outcomes.

3D-validation of a simple tool to measure tibiofemoral axial rotation in tibial plateau fractures.

OBJECTIVES: Rotated tibial plateau fractures (TPF) frequently involve multiple planes of movement, yet current presurgical assessment methods do not account for tibiofemoral axial rotation. This study introduces and validates a simple tool to measure rotation-the Gerdy-Tibial-Tuberosity-Surgical-Epicondylar-Axis (GTT-SEA) angle. METHODS: Forty-seven preoperative 2D CT from a TPF database at a tertiary trauma center were retrieved, and 3D models reconstructed. Three observers made repeated 2D and 3D measurements of the GTT-SEA angle, spaced 4 weeks apart, for 20 patients. Inter- and intra-observer agreement and 2D-3D correlation were calculated. A reference angle was defined from non-operated patients, to classify 28 patients with MRI into neutral, external rotation, and internal rotation groups. The classification agreement and soft tissue involvement between groups were analyzed. RESULTS: Mean 2D GTT-SEA angle was 17.65 ± 2.36° in non-operated patients, and 13.86 ± 3.90° in operated patients. 3D GTT-SEA angle was 18.92 ± 4.53° in non-operated patients, and 14.76 ± 6.03° in operated patients. 2D-3D correlation was moderate to good (ICC 0.64 ~ 0.83). Two-dimensional (ICC 0.70) and 3D (ICC 0.55) inter-observer agreements were moderate; 2D (ICC 0.82 ~ 0.88) and 3D (ICC 0.76 ~ 0.95) intra-observer agreements were good to excellent. Rotation classification agreement was slight (kappa 0.17) for 2D and good (kappa 0.76) for 3D. More popliteofibular ligament injury was detected in rotated knees (p = 0.016). CONCLUSIONS: The GTT-SEA angle offers simple, accessible, yet reliable measurement of tibiofemoral axial rotation. Though a true reference range remains to be determined, this tool adds valuable information to existing TPF classifications, potentially allowing assessment of soft tissue involvement in TPF. CLINICAL RELEVANCE STATEMENT: The GTT-SEA angle will benefit patients who sustain tibial plateau fractures, by allowing physicians to more accurately measure and plan for the injury in 3D, and raising suspicion for otherwise undetected soft tissue injuries, which can impact operative outcomes. KEY POINTS: • Traumatic fractures of the tibial plateau may contain rotation-induced soft tissue injuries. • A new tool to measure axial rotation between the femur and tibia was found to have moderate to excellent inter- and intra-rater reliability. • The tool may have potential in predicting soft tissue injury and assisting with the decision to receive MRI.

Targeting the elevated IFN-γ in vitiligo patients by human anti- IFN-γ monoclonal antibody hampers direct cytotoxicity in melanocyte.

BACKGROUND: Vitiligo is an autoimmune disease that progressively destroys melanocytes in the skin, resulting in patchy disfiguring depigmentation. The direct pathological effect of IFN-γ, CXCL10 to the melanocytes in vitiligo has been reported, but there are contradictory results to which cytokine exerts the critical cytotoxic effect on melanocytes. OBJECTIVE: The overarching goal was to study the direct toxicity of highly expressed cytokine in vitiligo skin lesions to melanocytes. METHODS: We obtained the interstitial fluid analyte from lesion and non-lesion skin of vitiligo patients and healthy control and sent for high sensitivity multiplex cytokine panel. We further performed functional study to identify the direct toxicity effect of the highly expressed cytokines. RESULTS: We found a significant elevation of IFN-γ, CXCL9, CXCL10, CXCL11 in the vitiligo skin. Ex vivo melanocyte studies support the direct role of IFN-γ per se in melanocyte cell loss, increased oxidative stress and melanogenesis disruption. Interestingly, we found that IFN-γ regulated cell death through oxidative stress-related ferroptosis cell death, which may initiate autoimmunity in vitiligo. In contrast to blocking selected cell death pathway, our in vitro study supports the rescue effect of human anti-IFN-γ monoclonal antibody 2A6Q to IFN-γ induced cell death, oxidative stress, and loss of function in melanocytes by interrupting IFN-γ signaling, which may be a potential therapeutic option for vitiligo. CONCLUSION: This study further confirms the direct of toxicity effect of IFN-γ per se towards melanocyte in vitiligo skin and the potential utility of human anti-IFN-γ monoclonal antibody in treating vitiligo.

The Stability of Hybrid Perovskites with UiO-66 Metal-Organic Framework Additives with Heat, Light, and Humidity.

This study is devoted to investigating the stability of metal-organic framework (MOF)-hybrid perovskites consisting of CH3NH3PbI3 (MAPbI3) and UiO-66 without a functional group and UiO-66 with different COOH, NH2,and F functional groups under external influences including heat, light, and humidity. By conducting crystallinity, optical, and X-ray photoelectron spectra (XPS) measurements after long-term aging, all of the prepared MAPbI3@UiO-66 nanocomposites (with pristine UiO-66 or UiO-66 with additional functional groups) were stable to light soaking and a relative humidity (RH) of 50%. Moreover, the UiO-66 and UiO-66-(F)4 hybrid perovskite films possessed a higher heat tolerance than the other two UiO-66 with the additional functional groups of NH2 and COOH. Tthe MAPbI3@UiO-66-(F)4 delivered the highest stability and improved optical properties after aging. This study provides a deeper understanding of the impact of the structure of hybrid MOFs on the stability of the composite films.

Cryptococcus gattii Infection as the Major Clinical Manifestation in Patients with Autoantibodies Against Granulocyte-Macrophage Colony-Stimulating Factor.

PURPOSE: Anti-granulocyte-macrophage colony-stimulating factor autoantibodies (anti-GM-CSF Abs) are a predisposing factor for pulmonary alveolar proteinosis (PAP) and Cryptococcus gattii cryptococcosis. This study aimed to investigate clinical manifestations in anti-GM-CSF Ab-positive patients with C. gattii cryptococcosis and analyze the properties of anti-GM-CSF Abs derived from these patients and patients with PAP. METHODS: Thirty-nine patients diagnosed with cryptococcosis (caused by C. neoformans or C. gattii) and 6 with PAP were enrolled in the present study. Clinical information was obtained from medical records. Blood samples were collected for analysis of autoantibody properties. We also explored the National Health Insurance Research Database (NHIRD) of Taiwan to investigate the epidemiology of cryptococcosis and PAP. RESULTS: High titers of neutralizing anti-GM-CSF Abs were identified in 15 patients with cryptococcosis (15/39, 38.5%). Most anti-GM-CSF Ab-positive cryptococcosis cases had central nervous system (CNS) involvement (14/15, 93.3%). Eleven out of 14 (78.6%) anti-GM-CSF Ab-positive CNS cryptococcosis patients were confirmed to be infected with C. gattii, and PAP did not occur synchronously or metachronously in a single patient from our cohort. Exploration of an association between HLA and anti-GM-CSF Ab positivity or differential properties of autoantibodies from cryptococcosis patients and PAP yielded no significant results. CONCLUSION: Anti-GM-CSF Abs can cause two diseases, C. gattii cryptococcosis and PAP, which seldom occur in the same subject. Current biological evidence regarding the properties of anti-GM-CSF Abs cannot provide clues regarding decisive mechanisms. Further analysis, including more extensive cohort studies and investigations into detailed properties, is mandatory to better understand the pathogenesis of anti-GM-CSF Abs.

Pathogenic autoantibodies to IFN-γ act through the impedance of receptor assembly and Fc-mediated response.

Anti-interferon (IFN)-γ autoantibodies (AIGAs) are a pathogenic factor in late-onset immunodeficiency with disseminated mycobacterial and other opportunistic infections. AIGAs block IFN-γ function, but their effects on IFN-γ signaling are unknown. Using a single-cell capture method, we isolated 19 IFN-γ-reactive monoclonal antibodies (mAbs) from patients with AIGAs. All displayed high-affinity (KD < 10-9 M) binding to IFN-γ, but only eight neutralized IFN-γ-STAT1 signaling and HLA-DR expression. Signal blockade and binding affinity were correlated and attributed to somatic hypermutations. Cross-competition assays identified three nonoverlapping binding sites (I-III) for AIGAs on IFN-γ. We found that site I mAb neutralized IFN-γ by blocking its binding to IFN-γR1. Site II and III mAbs bound the receptor-bound IFN-γ on the cell surface, abolishing IFN-γR1-IFN-γR2 heterodimerization and preventing downstream signaling. Site III mAbs mediated antibody-dependent cellular cytotoxicity, probably through antibody-IFN-γ complexes on cells. Pathogenic AIGAs underlie mycobacterial infections by the dual blockade of IFN-γ signaling and by eliminating IFN-γ-responsive cells.

Anti-interferon-γ autoantibody-associated immunodeficiency.

Anticytokine autoantibodies are an emerging disease etiology, through the disturbance of physiological functions of cognate cytokines. Anti-interferon (IFN)-γ autoantibodies (AIGAs) were first identified in patients with severe mycobacterial infections, and were considered to be an autoimmune phenocopy of inborn genetic errors of the IL-12/IFN-γ axis. More than 600 reported cases, most originating from Southeast Asia, have been diagnosed over the last decade. Specific HLA class II molecules are associated with these autoantibodies, which provide a genetic basis for the high prevalence of this immunodeficiency syndrome in certain ethnic groups. Salmonellosis and herpes zoster reactivation are observed in more than half the patients with AIGAs. Moreover, AIGAs have been shown to underlie severe Taralomyce marneffei infection in HIV-negative patients. AIGAs may, thus, be considered a new form of late-onset immunodeficiency conferring a predisposition not only to severe mycobacterial, but also to some bacterial and fungal infections.

Development of Compound Fault Diagnosis System for Gearbox Based on Convolutional Neural Network.

Gear transmission is widely used in mechanical equipment. In practice, if the gearbox is damaged, it not only affects the yield rate but also damages other parts of machines; thus, increases the cost and difficulty of maintenance. With the advancement of technology, the concept of unmanned factories has been proposed; an automatic diagnosis system for the health management of gearboxes becomes necessary. In this paper, a compound fault diagnosis system for the gearbox based on convolutional neural network (CNN) is developed. Specifically, three-axis vibration signals measured by accelerometers are used as the input of the one-dimensional CNN; the detection of the existence and type of the fault is directly output. In testing, the model achieved nearly 100% accuracy on the fault samples we captured. Experimental evidence also shows that the frequency-domain data can provide better diagnostic results than the time-domain data due to the stable characteristics in the frequency spectrum. For practical usage, we demonstrated a remote fault diagnosis system through a local area network on an embedded platform. Furthermore, optimization of convolution kernels was also investigated. When moderately reducing the number of convolution kernels, it does not affect the diagnostic accuracy but greatly reduces the training time of the model.

Synthesis of MOF525/PEDOT Composites as Microelectrodes for Electrochemical Sensing of Dopamine.

Dopamine (DA) is an important neurotransmitter responsible for the functions and activities of multiple systems in human. Electrochemical detection of DA has the advantages of fast analysis and cost-effectiveness, while a regular electrode probe is restricted to laboratory use because the probe size is too large to be suitable for an in vivo or in vitro analysis. In this study, we have developed porphyrin-based metal organic framework (MOF525) and poly(3,4-ethylenedioxythiophene) (PEDOT)-based composites to modify microelectrode for DA detection. Two types of PEDOT monomers with different functional groups were investigated in this study. By varying the monomer ratios, electrolyte concentrations, and electropolymerization temperature, it was found that the PEDOT monomer containing carboxylic group facilitated the formation of regular morphology during the electropolymerization process. The uniform morphology of the PEDOT promoted the electron transmission efficiency in the same direction, while the MOF525 provided a large reactive surface area for electrocatalysis of DA. Thus, the MOF525/PEDOT composite improved the sensitivity-to-noise ratio of DA signaling, where the sensitivity reached 11 nA/µM in a good linear range of 4-100 µM. In addition, porphyrin-based MOF could also increase the selectivity to DA against other common clinical interferences, such as ascorbic acid and uric acid. The as-synthesized microelectrode modified with MOF525/PEDOT in this study exhibited great potential in real time analysis.

Anti-IFN-γ autoantibodies underlie disseminated Talaromyces marneffei infections.

Talaromyces marneffei causes life-threatening opportunistic infections, mainly in Southeast Asia and South China. T. marneffei mainly infects patients with human immunodeficiency virus (HIV) but also infects individuals without known immunosuppression. Here we investigated the involvement of anti-IFN-γ autoantibodies in severe T. marneffei infections in HIV-negative patients. We enrolled 58 HIV-negative adults with severe T. marneffei infections who were otherwise healthy. We found a high prevalence of neutralizing anti-IFN-γ autoantibodies (94.8%) in this cohort. The presence of anti-IFN-γ autoantibodies was strongly associated with HLA-DRB1*16:02 and -DQB1*05:02 alleles in these patients. We demonstrated that adult-onset acquired immunodeficiency due to autoantibodies against IFN-γ is the major cause of severe T. marneffei infections in HIV-negative patients in regions where this fungus is endemic. The high prevalence of anti-IFN-γ autoantibody-associated HLA class II DRB1*16:02 and DQB1*05:02 alleles may account for severe T. marneffei infections in Southeast Asia. Our findings clarify the pathogenesis of T. marneffei infection and pave the way for developing novel treatments.

Dielectric Spectroscopy of Water Dynamics in Functionalized UiO-66 Metal-Organic Frameworks.

We present a dielectric spectroscopy study of dipolar dynamics in the hydrated UiO-66(Zr) type metal-organic frameworks (MOFs) functionalized with -NH2 and -F groups. Experiments are performed in a broad temperature and frequency ranges allowing us to probe several dipolar relaxations. For both samples at temperature below 220 K, we observe confined supercooled water dynamics, which can be described by the Arrhenius law. At slightly higher temperature, a second less pronounced dipolar relaxation is identified, and its origin is discussed. At even higher temperature, the dielectric permittivity exhibits anomalous increase with increasing temperature due to the proton conductivity. Upon further heating, the permittivity shows a sudden decrease indicating a reversible removal of water molecules. Measurements of the dehydrated samples reveal absence of all three dipolar processes.

HAI-2 as a novel inhibitor of plasmin represses lung cancer cell invasion and metastasis.

BACKGROUND: Dysregulation of pericellular proteolysis usually accounts for cancer cell invasion and metastasis. Isolation of a cell-surface protease system for lung cancer metastasis is an important issue for mechanistic studies and therapeutic target identification. METHODS: Immunohistochemistry of a tissue array (n = 64) and TCGA database (n = 255) were employed to assess the correlation between serine protease inhibitors (SPIs) and lung adenocarcinoma progression. The role of SPI in cell motility was examined using transwell assays. Pulldown and LC/MS/MS were performed to identify the SPI-modulated novel protease(s). A xenografted mouse model was harnessed to demonstrate the role of the SPI in lung cancer metastasis. RESULTS: Hepatocyte growth factor activator inhibitor-2 (HAI-2) was identified to be downregulated following lung cancer progression, which was related to poor survival and tumour invasion. We further isolated a serum-derived serine protease, plasmin, to be a novel target of HAI-2. Downregulation of HAI-2 promotes cell surface plasmin activity, EMT, and cell motility. HAI-2 can suppress plasmin-mediated activations of HGF and TGF-ß1, EMT and cell invasion. In addition, downregulated HAI-2 increased metastasis of lung adenocarcinoma via upregulating plasmin activity. CONCLUSION: HAI-2 functions as a novel inhibitor of plasmin to suppress lung cancer cell motility, EMT and metastasis.

Disseminated Cryptococcosis Due to Anti-Granulocyte-Macrophage Colony-Stimulating Factor Autoantibodies in the Absence of Pulmonary Alveolar Proteinosis.

INTRODUCTION: Autoantibodies to granulocyte-macrophage colony-stimulating factor (GM-CSF) can cause acquired pulmonary alveolar proteinosis (PAP). Cases of acquired PAP susceptible to typical respiratory pathogens and opportunistic infections have been reported. Anti-GM-CSF autoantibodies have been reported in a few patients with cryptococcal meningitis. This study evaluated the presence of neutralizing anti-GM-CSF autoantibodies in patients without known congenital or acquired immunodeficiency with severe pulmonary or extrapulmonary cryptococcal infection but without PAP. METHODS: We took a clinical history and performed an immunologic evaluation and screening of anti-cytokine autoantibodies in patients with cryptococcal meningitis. The impact of autoantibodies to GM-CSF on immune function was assessed by intracellular staining of GM-CSF-induced STAT5 phosphorylation and MIP-1α production in normal peripheral blood mononuclear cells incubated with plasma from patients or normal control subjects. RESULTS: Neutralizing anti-GM-CSF autoantibodies were identified in four patients with disseminated cryptococcosis, none of whom exhibited PAP. Plasma from patients blocked GM-CSF signaling and inhibited STAT5 phosphorylation and production of MIP-1α. One patient died of disseminated cryptococcosis involving the central nervous system, which was associated with defective GM-CSF activity. CONCLUSIONS: Anti-GM-CSF autoantibodies increase susceptibility to cryptococcal infection in adults without PAP. Cryptococcal central nervous system infection associated with anti-GM-CSF autoantibodies could result in neurological sequelae or be life-threatening. Therefore, timely detection of neutralizing anti-GM-CSF autoantibodies and development of an effective therapy are necessary to prevent deterioration of cryptococcal infection in these patients.

Identification of a major epitope by anti-interferon-γ autoantibodies in patients with mycobacterial disease.

The binding of autoantibodies (autoAbs) to interferon (IFN)-γ in people with mycobacterial diseases has become an emerging medical concern. Many patients display specific human leukocyte antigen (HLA) class II haplotypes, which suggests that a common T cell-dependent and B cell-dependent mechanism might underlie the production of specific anti-IFN-γ autoAbs. We show here that these autoAbs target a major epitope (amino acids 121-131, designated position (P)121-131) in a region crucial for IFN-γ receptor (IFN-γR) activation to impair IFN-γ-mediated activities. The amino acid sequence of this epitope is highly homologous to a stretch in the Noc2 protein of Aspergillus spp., which was cross-reactive with autoAbs from patients. Rats immunized with Aspergillus Noc2 developed antibodies that reacted with human IFN-γ. We generated an epitope-erased variant of IFN-γ (EE-IFN-γ), in which the major neutralizing epitope region was altered. The binding affinity of anti-IFN-γ autoAbs for EE-IFN-γ was reduced by about 40%, as compared to that for IFN-γ1-131. Moreover, EE-IFN-γ activated the IFN-γR downstream signaling pathway ex vivo, irrespectively of anti-IFN-γ autoAbs. In conclusion, we identified a common, crucial B cell epitope that bound to anti-IFN-γ autoAbs in patients, and we propose a molecular-mimicry model for autoAb development. In addition, treatment with EE-IFN-γ might be worth investigating in patients producing anti-IFN-γ autoAbs.

Clinical manifestations, course, and outcome of patients with neutralizing anti-interferon-γ autoantibodies and disseminated nontuberculous mycobacterial infections.

Neutralizing anti-interferon-γ autoantibody (nAIGA)-associated immunodeficiency is an emerging medical issue worldwide. In the present study, we describe and discuss the clinical features and outcomes of patients with nAIGAs and disseminated infections by nontuberculous mycobacteria (dNTM).We thoroughly reviewed the medical records of all patients. Microorganisms and nAIGAs were identified using previously described methods with modifications. All data were calculated and analyzed using SPSS software.Among 46 adult patients with dNTM infections, we identified 45 cases (97.8%) with nAIGAs. The average patient age was 58.6 years, and there was no sex predominance. Cervical lymphadenitis (81.8%) was the most common clinical manifestation. Endocrine disorder was the leading comorbidity (7 cases). Malignancies were found in 4 patients, and all of the malignancies originated from the T-cell/macrophage lineage. More than half of the identifiable isolates were slow-growing NTMs. Twenty-eight (62.2%) and 18 (40.0%) patients had a history of zoster and salmonellosis, respectively. A high proportion of patients with recurrent episodes of NTM infection or a history of zoster and dNTM infection had initial nAIGA titers ≥10 dilution (P < 0.05). Twenty-seven patients (60.0%) required long-term antimycobacterial therapy and had at least 1 episode of recurrent NTM disease. No mortality was related to dNTM infection.In Taiwan, nAIGAs are a recently recognized mechanism of dNTM infection. Long term of antibiotic treatment and adherence to medical advice are necessary to improve the clinical outcome of patients with nAIGAs.

Areca nut extracts exert different effects in oral cancer cells depending on serum concentration: A clue to the various oral alterations in betel quid chewers.

Betel quid chewing is associated with various pathologic alterations in oral mucosa. However, the molecular mechanism behind so many contradictory alterations remains unclear. Here we aimed to build a model to facilitate the related studies in cultured cells. In our results, areca nut extract (ANE) was found to exert different effects in oral cells depending on the supplemented serum level. ANE strongly induced DNA damage, necrotic ballooning, and inflammatory cytokines under lower serum concentration while might convert to facilitate deregulated growth of serum-supplemented cells via modulating the activity/expression of factors such as E-cadherin and Snail. Despite ANE significantly activated NF-κB, a mediator critical for inflammation, inhibition of NF-κB did not prevent the activation of IL8 promoter. We further discovered Y705-dephosphorylated STAT3 might enhance IL8 transcription. Since necrosis and the inflammatory cytokines could cause massive inflammation, infiltration of interstitial fluid might potentiate cellular resistance against the acute cytotoxicity of ANE and further support the proliferation of transforming cells. Induction of VEGF and angiogenesis under lower serum condition also paved the way for cell growth and subsequent metastasis. Accordingly, we concluded that in correlation with serum infiltration ANE caused particular effects in oral cells and possibly the various clinicopathological alterations in vivo.

Advanced oxidation of bromide-containing drinking water: a balance between bromate and trihalomethane formation control.

Addition of H202 has been employed to repress bromate formation during ozonation of bromide-containing source water. However, the addition of H2O2 will change the oxidation pathways of organic compounds due to the generation of abundant hydroxyl radicals, which could affect the removal efficacy of trihalomethane precursors via the combination of ozone and biological activated carbon (O3-BAC). In this study, we evaluated the effects of H2O2 addition on bromate formation and trihalomethane formation potential (THMFP) reduction during treatment of bromide-containing (97.6-129.1 microg/L) source water by the O3-BAC process. At an ozone dose of 4.2 mg/L, an H2O2/O3 (g/g) ratio of over 1.0 was required to maintain the bromate concentration below 10.0 microg/L, while a much lower H2O2/O3 ratio was sufficient for a lower ozone dose. An H2O2/O3 (g/g) ratio below 0.3 should be avoided since the bromate concentration will increase with increasing H2O2 dose below this ratio. However, the addition of H202 at an ozone dose of 3.2 mg/L and an H2O2/O3 ratio of 1.0 resulted in a 43% decrease in THMFP removal when comparedwith the O3-BAC process. The optimum H2O2/O3 (g/g) ratio for balancing bromate and trihalomethane control was about 0.7-1.0. Fractionation of organic materials showed that the addition of H2O2 decreased the removal efficacy of the hydrophilic matter fraction of DOC by ozonation and increased the reactivity of the hydrophobic fractions during formation of trihalomethane, which may be the two main reasons responsible for the decrease in THMFP reduction efficacy. Overall, this study clearly demonstrated that it is necessary to balance bromate reduction and THMFP control when adopting an H2O2 addition strategy.

Development of HPLC and NACE methods for the simultaneous determination of benzoic and sorbic acids in sour snap beans containing oil.

The practical methods were developed for the simultaneous determination of benzoic acid (BA) and sorbic acid (SA) in sour snap bean samples containing oil. BA and SA in the samples were extracted by ultrasonication with water, followed by cleanup procedures with precipitation for removing the potential proteins and with petroleum ether liquid-liquid extraction for removing the edible oil contained in the samples. The HPLC method was developed using Supelco C18 (250 mm x 4.6 mm id, 5 microm) as column, MeOH-20 mM NH(4)Ac (25:75 v/v) at 1.0 mL/min as the mobile phase and 230 nm as the detection wavelength. The optimal NACE method was established with a running buffer of 20.0 mM NH(4)Ac in 95% MeOH (pH* 10.6), and an applied voltage of -30 kV over a capillary of 50 microm id x 48.5 cm (40 cm to the detector window), which gave a baseline separation of BA and SA, and as well as of the blank matrix within ca. 10 min. Both HPLC and NACE methods gave the relatively lower limits of quantification at about 0.01-0.02 and 0.04-0.05 mg/kg, respectively, whereas the overall recoveries were larger than 85.0%. The proposed methods have been successfully applied to measure 15 real sour bean samples and the content profile of BA and SA in sour bean samples was obtained and evaluated.

Development of a NACE method for simultaneous measurement of three adenosine monophosphate isomers in biomimicking prebiotic synthesis without sample pretreatment.

A practical NACE method was developed for simultaneous determination of three adenosine monophosphate (AMP) isomers. Separation of three AMP isomers was achieved using 200 mM Tris/H(3)BO(3) in acetontrile/water (2:1 v/v) at pH* 10.0 as the running buffer and +25 kV as the applied voltage over a bare fused-silica capillary of 50 microm id x 375 microm od x 54.5 cm (46 cm to the detector window). At 260 nm, the calibration curves were linear in the range of 1-100 microg/mL. The detection limits were less than 0.70 microg/mL. The recovery ranged from 94.5 to 106.4%. The intraday RSDs of the migration times were between 2.1 and 3.0%. The developed NACE method has been successfully applied for the determination of three AMP isomers in the real samples of biomimicking prebiotic synthesis reaction between N-(O,O-diisopropyl) phosphoryl amino acid and adenosine.