High LC Research Articles

Clinical analysis of EBRT vs TLM in the treatment of early (T1-T2N0) glottic laryngeal cancer.

Objective: To analyze the clinical efficacy of external beam radiation therapy (EBRT) vs transoral laser microsurgery (TLM) in patients with early glottic laryngeal carcinoma (T1-T2N0) and the effect of treatment choice on vocal function.Methods: A retrospective analysis of patients with T1-T2N0 glottic laryngeal carcinoma who underwent EBRT or TLM between January 2012 and December 2018 in PUMCH. The Kaplan-Meier method was used to analyze local control, progression-free survival and overall survival, and the VHI-30 scale was used to evaluate the effects of EBRT and TLM on vocal function.Results: A total of 185 patients, all with pathologically confirmed squamous cell carcinoma, were enrolled. The median age was 62 years (38-88). N0 disease was confirmed by imaging: 142/185 (76.76%) patients had T1N0 disease, and 43/185 patients (23.24%) had T2/N0 disease. A total of 91/195 (49.19%) patients received an EBRT dose of 66-70 Gy/30-35f, at 2.0-2.3 Gy/f. 94/185 (50.81%) patients received TLM. The median follow-up time was 42 months (12-92), and the 3-year LC, PFS, and OS rates for the EBRT and TLM groups were 96.9% vs 94.1%(p=0.750), 95.3% vs 93.1%(p=0.993) and 93.3% vs 95.4%(p=0.467), respectively. The VHI-30 scales were used at the baseline showed no significant difference between the two groups 19.20±3.324 vs 21.65±9.80 (p=0.250), but the EBRT group had a low voice handicap after treatment, 10.24±6.093 vs 19.45±5.112 (p=0.001) (6 months) and 9.45±5.112 vs 14.97±7.741 (12 months). No CTCAE grade 3 or above side effects were observed in the EBRT group, but 3 cases of vocal cord stenosis were observed in the TLM group.Conclusion: The application of EBRT for early glottic laryngeal carcinoma (T1-T2N0) had an obvious curative effect with high LC and OS rates, no serious side effects, and a low voice handicap rate.

Investigation of the Causal Association between Long-Chain n-6 Polyunsaturated Fatty Acid Synthesis and the Risk of Type 2 Diabetes: A Mendelian Randomization Analysis

Background: Globally, 1 in 11 adults has diabetes mellitus, and most of these cases are type 2 diabetes (T2D). The risk of T2D is influenced by many factors, including diet. The synthesis of long-chain n-6 polyunsaturated fatty acids (LC n-6 PUFA) has been posited as a risk factor for T2D; however, its causal role is uncertain. Aim: To test the causal effect of LC n-6 PUFA synthesis on insulin resistance and transgenerational T2D risk in a large cohort of men and women. Methods: Two-sample mendelian randomization (MR) was conducted to evaluate the effect of low or high levels of LC n-6 PUFA synthesis on glycemia and development of T2D in the UK Biobank (n = 463,010) and Meta-Analysis of Glucose- and Insulin-Related Traits Consortium (MAGIC; n = 5,130) cohorts. The increased likelihood of a predisposition to low or high LC n-6 PUFA synthesis and the risk of T2D was also investigated using the participants’ siblings and parents. In MR-Base, 4 genetic variants associated with LC n-6 PUFA synthesis were found (p < 10^–8). After pruning, 1 variant (rs174547) on the FADS1 gene was retained. Results: Lower LC n-6 PUFA synthesis and abundance (per % unit decrease) are associated with small reductions in the insulin disposition index (–0.038 ± 0.012 mM^–1; p = 0.002) within MAGIC. In the UK Biobank, we report negligible effects of low n-6 PUFA synthesis on the odds of T2D (OR <1%; p < 0.05). Additionally, reduced LC n-6 PUFA synthesis does not appear to be a contributor to familial T2D risk. No significant association was observed between LC n-6 PUFA synthesis and BMI. Conclusion: In a primarily white European population, LC n-6 PUFA synthesis is not a major contributor to T2D risk.

High Even-Modulus Injection-Locked Frequency Dividers

This article designs and analyzes wide locking range (LR) high even-modulus LC -tank injection-locked frequency dividers (ILFDs) with current-reused topologies. The current-reused LC ILFD uses two stacked LC sub-ILFDs sharing the same dc current. The current-reused ILFD becomes a divide-by-4 ( $\div 4$ ) ILFD, when both sub-ILFDs are used $\div 2$ ILFDs, and it is used as a divide-by-8 ( $\div 8$ ) ILFD when one sub-ILFD is used a $\div 4$ ILFD. Both sub-ILFDs use nMOSFETs as linear injection mixers for high conversion gain. For the $\div 8$ LC ILFD designed in the TSMC 0.18- $\mu \text{m}$ CMOS process, the circuit uses one high-frequency $\div 4$ sub-ILFD and one low-frequency $\div 2$ sub-ILFD, at the supply of 1.6 V, and at the incident power of 0 dBm, the LR is 4 GHz (38.835%), from the incident frequency 8.3 to 12.3 GHz. The $\div 8$ ILFD core power consumption is 13.98 mW, and the die size is $1.2 \times 1.2$ mm2. Both the $\div 8$ LC ILFD and the $\div 4$ sub-ILFD have nonoverlapped and overlapped LRs, which are due to a dual-resonance resonator used in the varactor-free n-core sub-ILFD. The LC dual-resonance resonator is due to parasitic capacitors in active FETs and on-chip spiral inductors and inductive elements, and it is used to get wide overlapped LR. Two $\div 4$ LC ILFDs inherent in the designed circuit are also studied.

A high resolution LC–MS targeted method for the concomitant analysis of 11 contraceptive progestins and 4 steroids

In the context of hormonal contraception and hormone replacement therapy (HRT), many women are exposed to exogenous hormones. Current use of hormonal contraception with combined ethinyl estradiol and different progestins bestows a breast cancer relative risk (RR) of 1.2- while combined HRT has a RR of 2. Although these exposures present an important public health issue, little is known about the effects of individual progestins on the breast and other tissues. Increasing availability of large scale biobanks, high throughput analyses and data management tools enable ever expanding, sophisticated population studies. In order to address the impact of distinct progestins on various health indicators, it is desirable to accurately quantify progestins in clinical samples.Here we have developed and validated a high resolution liquid chromatography mass spectrometry (LC–MS) targeted method for the simultaneous quantification of 11 synthetic progestins widely used in oral contraceptives, gestodene, levonorgestrel, etonogestrel, chlormadinone acetate, cyproterone acetate, drospirenone, desacetyl norgestimate, medroxyprogesterone acetate, norethindrone, dienogest, nomegestrol acetate, and 4 endogenous steroid hormones, progesterone, testosterone, androstenedione, and cortisol in blood samples. This highly specific quantitative analysis with high resolution Orbitrap technology detects and quantifies 15 compounds using their internal standard counterparts in a single 12 min LC–MS run. Sensitivity is attained by the use of the instrument in targeted selected ion monitoring mode. Lower limit of quantitation ranges from 2.4 pg/ml for drospirenone to 78.1 pg/ml for chlormadinone acetate. The method provides comprehensive progestin panel measurements with as little as 50 μl of murine or human plasma.

Rational selection of reverse phase columns for high throughput LC-MS lipidomics.

Natural lipidomes are characterized by extremely high complexity and dynamic range of lipid concentrations. Furthermore, high diversity of lipid physicochemical properties requires high resolving powers for both chromatographic and mass spectrometric analytical platforms. Reverse-phase chromatography coupled with data-dependent MS/MS acquisition is one of the most popular techniques in untargeted lipidomics. Optimal method should provide good chromatographic separation and resolution, reproducibility, selectivity and sensitivity. Here, we developed and set-up a RPLC-MS/MS workflow capable of resolving complex mixtures of lipids in 32 min of analysis. Human blood plasma was chosen as a representative complex natural lipidome with large variance of lipid classes, species and lipid concentrations. Lipids were separated by RPLC on five different reverse phase columns with different types of stationary phase particles, size and chemistry. High mass accuracy MS analysis and data-dependent MS/MS analysis were performed using a Q Exactive™ HF Hybrid Quadrupole-Orbitrap™ Mass Spectrometer to identify individual lipid molecular species. This workflow was applied to evaluate the separation capability of each column and to identify the lipidomics profile in highly complex biological samples. As a result, we report more than 600 lipid species covering 18 lipid classes in human blood plasma and provide suggestions to the selection of the appropriate reverse phase column for the analysis of specific lipidomes.

Synovial fluid proteome changes in ACL injury-induced posttraumatic osteoarthritis: Proteomics analysis of porcine knee synovial fluid.

Surgical transection of the anterior cruciate ligament (ACL) in the porcine model leads to posttraumatic osteoarthritis if left untreated. However, a recently developed surgical treatment, bridge-enhanced ACL repair, prevents further cartilage damage. Since the synovial fluid bathes all the intrinsic structures of knee, we reasoned that a comparative analysis of synovial fluid protein contents could help to better understand the observed chondroprotective effects of the bridge-enhanced ACL repair. We hypothesized that post-surgical changes in the synovial fluid proteome would be different in the untreated and repaired knees, and those changes would correlate with the degree of cartilage damage. Thirty adolescent Yucatan mini-pigs underwent unilateral ACL transection and were randomly assigned to either no further treatment (ACLT, n = 14) or bridge-enhanced ACL repair (BEAR, n = 16). We used an isotopically labeled high resolution LC MS/MS-based proteomics approach to analyze the protein profile of synovial fluid at 6 and 12 months after ACL transection in untreated and repaired porcine knees. A linear mixed effect model was used to compare the normalized protein abundance levels between the groups at each time point. Bivariate linear regression analyses were used to assess the correlations between the macroscopic cartilage damage (total lesion area) and normalized abundance levels of each of the identified secreted proteins. There were no significant differences in cartilage lesion area or quantitative abundance levels of the secreted proteins between the ACLT and BEAR groups at 6 months. However, by 12 months, greater cartilage damage was seen in the ACLT group compared to the BEAR group (p = 0.005). This damage was accompanied by differences in the abundance levels of secreted proteins, with higher levels of Vitamin K-dependent protein C (p = 0.001), and lower levels of Apolipoprotein A4 (p = 0.021) and Cartilage intermediate layer protein 1 (p = 0.049) in the ACLT group compared to the BEAR group. There were also group differences in the secreted proteins that significantly changed in abundance between 6 and 12 months in ACLT and BEAR knees. Increased concentration of Ig lambda-1 chain C regions and decreased concentration of Hemopexin, Clusterin, Coagulation factor 12 and Cartilage intermediate layer protein 1 were associated with greater cartilage lesion area. In general, ACLT knees had higher concentrations of pro-inflammatory proteins and lower concentrations of anti-inflammatory proteins than BEAR group. In addition, the ACLT group had a lower and declining synovial concentrations of CILP, in contrast to a consistently high abundance of CILP in repaired knees. These differences suggest that the knees treated with bridge-enhanced ACL repair may be maintaining an environment that is more protective of the extracellular matrix, a function which is not seen in the ACLT knees.

Somatic proteome of Haemonchus contortus

Currently, there is a dearth of proteomic data to underpin fundamental investigations of parasites and parasitism at the molecular level. Here, using a high throughput LC–MS/MS-based approach, we undertook the first reported comprehensive, large-scale proteomic investigation of the barber's pole worm (Haemonchus contortus) – one of the most important parasitic nematodes of livestock animals worldwide. In total, 2487 unique H. contortus proteins representing different developmental stages/sexes (i.e. eggs, L3s and L4s, female (Af) and male (Am) adults) were identified and quantified with high confidence. Bioinformatic analyses of this proteome revealed substantial alterations in protein profiles during the life cycle, particularly in the transition from the free-living to the parasitic phase, and key groups of proteins involved specifically in feeding, digestion, metabolism, development, parasite-host interactions (including immunomodulation), structural remodelling of the body wall and adaptive processes during parasitism. This proteomic data set will facilitate future molecular, biochemical and physiological investigations of H. contortus and related nematodes, and the discovery of novel intervention targets against haemonchosis.

Characterization of the plasma proteome of nonhuman primates during Ebola virus disease or melioidosis: a host response comparison

BackgroundIn-depth examination of the plasma proteomic response to infection with a wide variety of pathogens can assist in the development of new diagnostic paradigms, while providing insight into the interdependent pathogenic processes which encompass a host’s immunological and physiological responses. Ebola virus (EBOV) causes a highly lethal infection termed Ebola virus disease (EVD) in primates and humans. The Gram negative non-spore forming bacillus Burkholderia pseudomallei (Bp) causes melioidosis in primates and humans, characterized by severe pneumonia with high mortality. We sought to examine the host response to infection with these two bio-threat pathogens using established animal models to provide information on the feasibility of pre-symptomatic diagnosis, since the induction of host molecular signaling networks can occur before clinical presentation and pathogen detection.MethodsHerein we report the quantitative proteomic analysis of plasma collected at various times of disease progression from 10 EBOV-infected and 5 Bp-infected nonhuman primates (NHP). Our strategy employed high resolution LC–MS/MS and a peptide-tagging approach for relative protein quantitation. In each infection type, for all proteins with > 1.3 fold abundance change at any post-infection time point, a direct comparison was made with levels obtained from plasma collected daily from 5 naïve rhesus macaques, to determine the fold changes that were significant, and establish the natural variability of abundance for endogenous plasma proteins.ResultsA total of 41 plasma proteins displayed significant alterations in abundance during EBOV infection, and 28 proteins had altered levels during Bp infection, when compared to naïve NHPs. Many major acute phase proteins quantitated displayed similar fold-changes between the two infection types but exhibited different temporal dynamics. Proteins related to the clotting cascade, immune signaling and complement system exhibited significant differential abundance during infection with EBOV or Bp, indicating a specificity of the response.ConclusionsThese results advance our understanding of the global plasma proteomic response to EBOV and Bp infection in relevant primate models for human disease and provide insight into potential innate immune response differences between viral and bacterial infections.

Reductive Desulfuration as an Important Tool in Detection of Small Molecule Modifications to Payload of Antibody Drug Conjugates.

A high resolution accurate mass LC-MS method was developed to facilitate the characterization of a subset of antibody drug conjugate (ADC) biotherapeutics, where the payload is linked to the antibody by a thioether bond. Desulfuration of the thioether linker was optimized for release of the payload to take advantage of the high resolution and high mass accuracy of the Orbitrap to characterize metabolism of the payload. Two model ADCs, trastuzumab emtansine (T-DM1) and SigmaMAb dansyl-cadavarine-SMCC (SigmaMAb ADC mimic) were selected for optimization of the desulfuration reaction as a function of reaction time, pH, organic solvent, and chaotropic reagents (urea, guanidine HCl) by monitoring the yield of released desulfurated DM1 from T-DM1 and desulfurated dansyl-cadaverine-SMCC from SigmaMAb ADC mimic, respectively. The optimized desulfuration technique was successfully applied to enable characterization of the ADC following its incubation in hepatocytes, liver microsomes, and buffers, as illustrated by the identification of a hydrolyzed thiosuccinimide ring of SigmaMAb ADC mimic following incubation in buffer for 48 h. The results from this study demonstrate that the chemical cleavage of thioether bond by desulfuration is simple, efficient, and specific. This technique is useful in characterization of metabolism on the payload of ADC to provide guidance for improvement of its biopharmaceutical profile. This is the first report on characterization of modification to payload of ADC following desulfuration.

Preparation of Human Skeletal Muscle Samples for Proteomic Analysis with Isobaric iTRAQ Labels

In the past decade, mass spectrometry studies of skeletal muscles have become common. In this tissue, the abundance of several contractile proteins significantly limits the depth of the panoramic proteome analysis. The use of isobaric labels allows improving assessment of the changes in the protein content, while analyzing up to 10 samples in a single run. Here we present the results of a comparative study of various methods for the fractionation of skeletal muscle peptides labeled with an isobaric label iTRAQ. Samples from m. vastus lateralis of eight young males were collected with a needle biopsy. After digestion into peptides and labeling, the preparations were carried out according to three different protocols: (1) peptide purification, HPLC-MS/MS; (2) peptide purification, isoelectric focusing, HPLC-MS/MS; (3) high pH reverse-phase LC fractionation, HPLC-MS/MS. Fractionation of labeled peptides by high pH reverse-phase LC was the optimal strategy for increasing the depth of the proteome analysis. This approach, in addition to contractile and mitochondrial proteins, allowed us to detect a variety of regulatory molecules, including the nucleic acids binding the proteins, chaperones, receptors, and transcription factors.

Phytochemical analysis of Holothuria leucospilota, a sea cucumber from Persian Gulf.

Sea cucumbers are widely consumed in traditional medicine and food. These animals have a considerable secondary metabolite and also several potential biological activities. This study investigated the phytochemical and cytotoxic evaluation of Holothuria leucospilota (H. leucospilota), a sea cucumber from Persian Gulf. The saponin composition of H. leucospilota was studied by different partitioning and chromatography methods such as thin layer chromatography (TLC), medium pressure liquid chromatography (MPLC) and high performance LC (HPLC). The marine sea cucumber Holostane-type triterpenoids (1-3) were characterized by physical and spectroscopic examination (1 and 2 dimensional neuclear magnetic resonance and mass experiments) with data analysis. The structure of compounds 1-3 identified as echinoside A, holothurine A, and 24-dehydroechinoside A, showed moderate cytotoxic activity with IC50 values of 1.9 ± 0.07, 6.8 ± 0.23, and 2.57 ± 0.18 μg/mL against HeLa and 10.4 ± 0.32, 8.9 ± 0.24, and 4.4 ± 0.13 on HUVEC cell line, respectively. In conclusion, the holostane-type triterpenoids showed moderate cytotoxic activity against HeLa cell line and have a prosperous future to be introduced as a lead structure.

High sensitive LC–MS/MS method for estimation of eluxadoline in human plasma and its application to pharmacokinetic study

A selective, sensitive and high throughput liquid chromatography – tandem mass spectrometry (LC–MS/MS) method was developed and validated for estimation of eluxadoline in human plasma. Plasma samples of analyte with internal standard (eluxadoline13CD3) were extracted using solid phase extraction on Orochem Panthera Deluxe cartridges. Chromatographic separation was performed on Ace Phenyl column (150 × 4.6 mm, 5 μm), using a mixture of buffer (2 mM ammonium acetate in 0.01% formic acid), acetonitrile and methanol (20:70:10, v/v/v) as mobile phase at a flow rate of 0.8 mL/min. The run time of analyte was 4.0 min. Tandem mass spectrometry, operating in positive ionization and multiple reaction monitoring modes was used for detection of analyte and internal standard. The method was established with a linear dynamic range of 25.0–5000 pg/mL for eluxadoline using 300 μL human plasma. The sample preparation procedure was consistent and reproducible (accuracy, 96.2–106.1%; precision (%CV), 0.8–6.6%), preventing the ex vivo hydrolysis of acyl glucuronide metabolite of eluxadoline to parent drug. The method was applied successfully to a clinical pharmacokinetic study in six healthy South Indian male subjects under fed conditions and the results were authenticated by incurred sample reanalysis.

An investigation of the molecular and biochemical basis underlying chlorantraniliprole-resistant Drosophila strains and their cross-resistance to other insecticides.

Chlorantraniliprole is an anthranilic diamide insecticide that binds to the insect ryanodine receptor (RyR) and induces an uncontrolled release of Ca2+ , resulting in paralysis and ultimately death of the target insects. Recently, it was reported that chlorantraniliprole-resistant diamondback moths, Plutella xylostella Linnaeus, have mutations in their RyR. In this study, we developed two different chlorantraniliprole-resistant Drosophila melanogaster strain. The resistance ratio (RR) of the low-concentration chlorantraniliprole-treated resistant (Low-Res) strain was 2.3, while that of the high-concentration chlorantraniliprole-treated resistant (High-Res) strain was 21.3. The LC 50 of the untreated control (Con) strain was 23.8~25.9 ppm, which was significantly higher than that reported for the susceptible diamondback moth (0.03~0.51 ppm). The high LC 50 of the Con may be because the helix S2 amino acid sequence of D. melanogaster RyR ( DmRyR) is identical to the I4790M mutation of the chlorantraniliprole-resistant diamondback moths, resulting in a lower binding affinity of DmRyR for chlorantraniliprole. Among the tested detoxification enzymes, the activity of esterase was significantly increased in the two Res strains, but glutathione S-transferases and acetylcholinesterase were significantly decreased in the two Res strains. The cross-resistance of the High-Res strain to other insecticides with different modes of actions (MoAs) revealed that the RRs of the neuronal acetylcholine receptor allosteric and competitive modulators were significantly increased, while those of the Na 2+ channel modulators were significantly reduced. Our studies showed that RRs against the same insecticide vary with the treatment concentration, and that RRs against other insecticides with different MoAs can be altered.

The physical and electrochemical properties of activated carbon electrode made from Terminalia Catappa leaf (TCL) for supercapacitor cell application

In this study, the utilization of TCL as raw material in production of carbon supercapacitor electrodes are focused on variations in carbonization temperature. Carbonization temperatures were varied by 500 °C, 600 °C, 700 °C, and 800 °C marked with the code TCL500, TCL600, TCL700 and TCL800 corresponding to the selection of carbonization temperature. All samples were activated using CO2 gas at 850 °C for 2 hours and 30 minutes. The density of the resulting carbon electrode was 0.776 gcm−3, 0.754 gcm−3, 0.738 gcm−3 and 0.762 gcm−3 for electrodes TCL500, TCL600, TCL700 and TCL800, respectively. The crystalline nature of the resulting carbon sample was found to be amorphous with 2θ 23.880°, 44.192° scattering angle, and stacking high (Lc) 8.482 Å for TCL700 samples. Lc data has been used to obtain a sample surface area and it was found as high as 1160.497 m2g−1. The morphology of the electrode surface has been tested using a SEM instrument. The elements content of the electrode have also been analyzed based on the EDS data. The optimum specific capacitance of carbon electrodes from TCL is obtained at a carbonization temperature of 700 °C as high as 54 Fg−1.

The developmental lipidome of Haemonchus contortus

Lipids play crucial roles in the biology of organisms, particularly relating to cellular membranes, energy storage, and intra- or inter-cellular signalling. Despite the recent expansion of the lipidomics field, very little is known about the biology of lipids in metzoan pathogens, and, to date, there has been no global lipidomic study of a parasitic nematode. Using Haemonchus contortus (barber's pole worm) as a model, we describe the first known global lipidome for a parasitic nematode via high throughput LC–MS/MS-based lipidomics. We identified a total of 554 lipid species across four lipid categories, and 18 lipid classes exhibited alterations among six developmental stages (eggs; L3 and exsheathed L3 (xL3) and L4 larval stages; female and male adults) of H. contortus. The lipid composition and abundance of H. contortus changed significantly during the transition from free-living (egg, L3 and xL3) to parasitic (L4 and adult) stages. The three main changes observed were: (i) decreased synthesis of triradylglycerols; (ii) increased glycerophospholipids (predominantly glycerophosphoethanolamines and glycerophosphocholines); and (iii) a ‘cooperative’ modulation of ether-linked lipids and saturated fatty acids. These changes suggest specific adaptations, in terms of nutrient acquisition, metabolism and development, as the nematode makes its transition to the parasitic stage inside the host animal. This lipidomic data set serves as a stimulus for studies to understand lipid biology in parasitic worms, and their roles in parasite–host interactions and disease processes.

Supercritical Fluid Chromatography × Ultra-High Pressure Liquid Chromatography for Red Chilli Pepper Fingerprinting by Photodiode Array, Quadrupole-Time-of-Flight and Ion Mobility Mass Spectrometry (SFC × RP-UHPLC-PDA-Q-ToF MS-IMS)

The coupling of normal phase and reversed phase liquid chromatography (NP-LC × RP-LC) is one of the most effective ways to increase orthogonality in two-dimensional comprehensive separations, being the two retention mechanisms truly independent. However, such coupling is not straightforward to implement, due to immiscibility of the mobile phases, poor peak focusing at the head of the secondary column (2D), signal interferences and peak deterioration. In this research, supercritical fluid chromatography (SFC) was used in the first dimension (1D), coupled to ultra high pressure LC (UHPLC), to alleviate incompatibility issues and provide a number of advantages related to the use of supercritical CO2. An on-line SFC × RP-UHPLC system was implemented in an automated fashion, around two 2-position, six-port switching valves equipped with octadecyl silica cartridges for effective trapping and focusing of the analytes eluted from 1D. A water make-up flow added to the SFC effluent permitted to efficiently focus the solutes on the sorbent material and to reduce interferences of the expanded CO2 on the 2D separation. In addition to photodiode array and quadrupole-time-of-flight mass spectrometry detection, ion mobility separation based on analyte mass, shape and size added a fourth separation dimension for carotenoid fingerprinting in a red chilli pepper sample.

A novel sample preparation procedure for effect-directed analysis of micro-contaminants of emerging concern in surface waters

A novel sample preparation procedure relying on Solid Phase Extraction (SPE) combining different sorbent materials on a sequential-based cartridge was optimized and validated for the enrichment of 117 widely diverse contaminants of emerging concern (CECs) from surface waters (SW) and further combined chemical and biological analysis on subsequent extracts. A liquid chromatography coupled to high resolution tandem mass spectrometry LC-(HR)MS/MS protocol was optimized and validated for the quantitative analysis of organic CECs in SW extracts. A battery of in vitro CALUX bioassays for the assessment of endocrine, metabolic and genotoxic interference and oxidative stress were performed on the same SW extracts. Satisfactory recoveries ([70–130]%) and precision (< 30%) were obtained for the majority of compounds tested. Internal standard calibration curves used for quantification of CECs, achieved the linearity criteria (r2 > 0.99) over three orders of magnitude. Instrumental limits of detection and method limits of quantification were of [1–96] pg injected and [0.1–58] ng/L, respectively; while corresponding intra-day and inter-day precision did not exceed 11% and 20%. The developed procedure was successfully applied for the combined chemical and toxicological assessment of SW intended for drinking water supply. Levels of compounds varied from < 10 ng/L to < 500 ng/L. Endocrine (i.e. estrogenic and anti-androgenic) and metabolic interference responses were observed. Given the demonstrated reliability of the validated sample preparation method, the authors propose its integration in an effect-directed analysis procedure for a proper evaluation of SW quality and hazard assessment of CECs.

Clinical outcomes of radio-hyperthermo-chemotherapy for soft tissue sarcoma compared to a soft tissue sarcoma registry in Japan: a retrospective matched-pair cohort study.

Regional hyperthermia is considered to enhance the antitumor effects of chemotherapy and radiotherapy. In this study, we confirmed the efficacy of concomitant radiotherapy, hyperthermia, and chemotherapy (RHC) for neoadjuvant treatment of malignant soft tissue sarcoma (STS). From 1994 to 2013, we performed RHC in 150 patients. This study was limited to 60 patients using the following exclusion criteria: salvage for recurrence or unplanned excision, trunk location, metastasis at initiation, non‐STS, and no definitive surgery. As a control group, we collected data from 11,031 patients in the Bone and Soft Tissue Tumor Registry in Japan (BSTT). We performed multivariate logistic regression analysis, and propensity scores were created for comparisons between groups. The primary outcome of this study was to compare oncologic outcomes (5‐year local control rate [LC] and overall survival rate [OS]). In the RHC group, two local recurrences (3.3%) occurred, and no patients underwent amputation. Margins of definitive surgery were not identical between groups [wide margins (60.0% vs. 85.3%), marginal margins (28.3% vs. 10.5%), and intralesional margins (7.4% vs. 4.2%), RHC and BSTT groups, respectively, P < 0.001]. After adjustment, the difference in OS was not significant between groups (HR = 1.26, P = 0.532); however, a statistically significant difference in LC was observed (HR = 4.82, P = 0.037). RHC resulted in a high LC at 5 years compared to the BSTT group, and amputation was averted in the RHC group, despite the wider margins in the BSTT group. This indicates that less invasive surgery might be achieved with effective neoadjuvant therapy.

Screening of 23 β-lactams in foodstuffs by LC–MS/MS using an alkaline QuEChERS-like extraction

ABSTRACTA fast and robust high performance LC–MS/MS screening method was developed for the analysis of β-lactam antibiotics in foods of animal origin: eggs, raw milk, processed dairy ingredients, infant formula, and meat- and fish-based products including baby foods. QuEChERS extraction with some adaptations enabled 23 drugs to be simultaneously monitored. Screening target concentrations were set at levels adequate to ensure compliance with current European, Chinese, US and Canadian regulations. The method was fully validated according to the European Community Reference Laboratories Residues Guidelines using 93 food samples of different composition. False-negative and false-positive rates were below 5% for all analytes. The method is adequate for use in high-routine laboratories. A 1-year study was additionally conducted to assess the stability of the 23 analytes in the working standard solution.

High Sensitivity and Precision High-Temperature Reversed-Phase LC Analysis of Bevacizumab for Intact Bioanalysis of Therapeutic Monoclonal Antibodies

High Sensitivity and Precision High-Temperature Reversed-Phase LC Analysis of Bevacizumab for Intact Bioanalysis of Therapeutic Monoclonal Antibodies