A multicenter, double-blind, randomized, parallel-group, active-controlled, phase 3 clinical trial to compare the effectiveness and safety of two botulinum toxin type A formulations for improving moderate to severe glabellar wrinkles in Asians.

Botulinum toxin type A (BoNT-A) was first isolated in 1946, and since then, several formulations have been developed and widely used to treat wrinkles by inducing muscle paralysis. This multicenter, double-blind, randomized, parallel-group, active-controlled phase 3 clinical trial was designed to evaluate the efficacy and safety of a newly developed BoNT-A formulation, BMI2006, in improving moderate to severe glabellar wrinkles and to compare with existing onabotulinumtoxin A (OBoNT) injections. A total of 276 subjects were enrolled and received 20 units of the randomized material, which was intramuscularly injected into five different locations on the forehead. The primary endpoint, assessed at 4 weeks, showed no statistically significant difference in the improvement rate of glabellar wrinkles between the two groups, with BMI2006 demonstrating non-inferiority to comparator BoNT-A. Secondary endpoints, evaluated by both treating investigators and independent investigators, also exhibited similar improvement rates throughout the study period. Both groups reported high levels of satisfaction with no statistical difference between the two groups. Safety evaluations indicated mild and transient adverse events, with no serious reactions observed. In conclusion, BMI2006 is an effective and safe BoNT-A for treating glabellar wrinkles with an expected duration of action between 8 and 12 weeks.

Lespedeza maximowiczii flower absolute promotes skin epithelization, barrier properties, and moisturization-related beneficial responses in human keratinocytes.

Lespedeza maximowiczii (LM), a member of the legume family, has tyrosinase inhibitory and estrogenic activities. However, its effects on skin-related biological activities remain unclear. Therefore, the present study aimed to explore the effects of LM flower absolute (LMFAb) on skin-related biological events, especially skin re-epithelization, barrier and moisturizing-related keratinocyte (HaCaT cell) responses. In this study, LMFAb was isolated from LM flowers via solvent extraction and its chemical composition analysis was performed using gas chromatography/mass spectrometry. 5-bromo-2'-deoxyuridine incorporation, Boyden chamber, sprout outgrowth, enzyme-linked immunosorbent, and Western blot assay were used to analyze the biological effects of LMFAb on HaCaT cells (a human epidermal keratinocyte cell line). Twelve components were identified in LMFAb. LMFAb promoted cell proliferation, migration, and sprout outgrowth in HaCaT cells. The absolute enhanced the activations of MAPKs (ERK1/2, JNK, and p38), PI3K and AKT proteins in HaCaT cells and elevated collagen type I and IV levels in HaCaT cell conditioned medium. In addition, LMFAb induced an increase in the expression levels of epidermal barrier proteins (filaggrin and involucrin) in HaCaT cells. Furthermore, LMFAb increased hyaluronan (HA) production and expression of HA synthases (HAS-1, HAS-2, and HAS-3) but decreased HYBID (HA binding protein involved in HA depolymerization) level in HaCaT cells. These findings demonstrate that LMFAb might promote skin re-epithelization, barrier and moisturizing-related beneficial responses in keratinocytes. This study suggests that LMFAb should be considered a potential starting material for the development of cosmetic or pharmaceutical agents that restore the functions of damaged skin.

Chemical Composition and Skin-Whitening Activities of Siegesbeckia glabrescens Makino Flower Absolute in Melanocytes.

Siegesbeckia glabrescens Makino (SGM) has been traditionally used to treat many disorders, including rheumatoid arthritis, hypertension, and acute hepatitis. However, the biological activities of SGM in skin remain unclear. The present study explored the effects of SGM flower absolute (SGMFAb) on skin-whitening-linked biological activities in B16BL6 cells. SGMFAb was extracted using hexane, and its composition was analyzed through gas chromatography/mass spectrometry analysis. The biological effects of SGMFAb on B16BL6 melanoma cells were detected via WST and BrdU incorporation assays, ELISA, and immunoblotting. SGMFAb contained 14 compounds. In addition, SGMFAb was noncytotoxic, attenuated the serum-induced proliferation of, and inhibited melanin synthesis and tyrosinase activity in α-MSH-exposed B16BL6 cells. SGMFAb also reduced the expressions of MITF (microphthalmia-associated transcription factor), tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2 in α-MSH-exposed B16BL6 cells. Moreover, SGMFAb downregulated the activation of p38 MAPK, ERK1/2, and JNK in α-MSH-stimulated B16BL6 cells. In addition, SGMFAb reduced the expressions of three melanosome-transport-participating proteins (myosin Va, melanophilin, and Rab27a) in α-MSH-stimulated B16BL6 cells. These results indicate that SGMFAb positively influences skin whitening activities by inhibiting melanogenesis and melanosome-transport-related events in B16BL6 cells, and suggest that SGMFAb is a promising material for developing functional skin whitening agents.

Analysis of the Microbiome of the Ear Canal in Normal Individuals and Patients with Chronic Otitis Externa.

BACKGROUND: Recently, microbiome research has been actively conducted for various skin areas. However, no study has yet compared the microbiome of bacteria and fungi in the ear canal of healthy individuals and patients with chronic otitis externa in Korea. OBJECTIVE: This study aimed to investigate the difference in the distribution of fungal and bacterial microbial communities in ear canal samples of healthy individuals and patients with chronic otitis externa. METHODS: In 24 patients with bilateral chronic otitis externa and 24 healthy controls, cotton swabs were used to obtain samples from the bilateral ear canal. To characterize the fungal and bacterial communities, we sequenced and analyzed the 16S rRNA V4-V5 and ITS1 regions using Quantitative Insights into Microbial Ecology 2, respectively. RESULTS: The alpha diversity analysis for bacteria and fungi confirmed that both richness and evenness decreased in the patient group. The beta diversity analysis for bacteria confirmed that these parameters differed between the control and patient groups. The beta diversity analysis for fungi showed no difference between the groups. CONCLUSION: We observed different skin microbiomes in the patients with chronic otitis externa compared with those in the healthy individuals.

Dynamic alterations in PD-1/PD-L1 expression level and immune cell profiles based on radiation response status in mouse tumor model.

Introduction: Based on the immunologic effects of anti-cancer treatment and their therapeutic implications, we evaluated radiotherapy (RT)-induced dynamic alterations in programmed death-1 (PD-1)/PD ligand-1 (PD-L1) expression profiles. Methods: Local RT with 2 Gy × 5 or 7.5 Gy × 1 was administered to the CT26 mouse model. Thereafter, tumors were resected and evaluated at the following predefined timepoints according to radiation response status: baseline, early (immediately after RT), middle (beginning of tumor shrinkage), late (stable status with RT effect), and progression (tumor regrowth). PD-1/PD-L1 activity and related immune cell profiles were quantitatively assessed. Results: RT upregulated PD-L1 expression in tumor cells from the middle to late phase; however, the levels subsequently decreased to levels comparable to baseline in the progression phase. RT with 2 Gy × 5 induced a higher frequency of PD-L1+ myeloid-derived suppressor cells, with a lesser degree of tumor regression, compared to 7.5 Gy. The proportion of PD-1+ and interferon (IFN)-γ+CD8α T cells continued to increase. The frequency of splenic PD-1+CD8+ T cells was markedly elevated, and was sustained longer with 2 Gy × 5. Based on the transcriptomic data, RT stimulated the transcription of immune-related genes, leading to sequentially altered patterns. Discussion: The dynamic alterations in PD-1/PD-L1 expression level were observed according to the time phases of tumor regression. This study suggests the influence of tumor cell killing and radiation dosing strategy on the tumor immune microenvironment.

PI3Kδ/γ inhibitor BR101801 extrinsically potentiates effector CD8+ T cell-dependent antitumor immunity and abscopal effect after local irradiation.

BACKGROUND: Radiotherapy enhances antitumor immunity. However, it also induces immunosuppressive responses, which are major hurdles for an effective treatment. Thus, targeting the immunosuppressive tumor microenvironment is essential for enhancing the antitumor immunity after radiotherapy. Retrospective studies show that a blockade of PI3Kδ and/or γ, which are abundant in leukocytes, exhibits antitumor immune response by attenuating activity of immune suppressive cells, however, the single blockade of PI3Kδ or γ is not sufficient to completely eliminate solid tumor. METHODS: We used BR101801, PI3Kδ/γ inhibitor in the CT-26 syngeneic mouse model with a subcutaneously implanted tumor. BR101801 was administered daily, and the target tumor site was locally irradiated. We monitored the tumor growth regularly and evaluated the immunological changes using flow cytometry, ELISpot, and transcriptional analysis. RESULTS: This study showed that BR101801 combined with irradiation promotes systemic antitumor immunity and abscopal response by attenuating the activity of immune suppressive cells in the CT-26 tumor model. BR101801 combined with irradiation systemically reduced the proliferation of regulatory T cells (Tregs) and enhanced the number of tumor-specific CD8α+ T cells in the tumor microenvironment, thereby leading to tumor regression. Furthermore, the high ratio of CD8α+ T cells to Tregs was maintained for 14 days after irradiation, resulting in remote tumor regression in metastatic lesions, the so-called abscopal effect. Moreover, our transcriptomic analysis showed that BR101801 combined with irradiation promoted the immune-stimulatory tumor microenvironment, suggesting that the combined therapy converts immunologically cold tumors into hot one. CONCLUSIONS: Our data suggest the first evidence that PI3Kδ/γ inhibition combined with irradiation promotes systemic antitumor immunity against solid tumors, providing the preclinical result of the potential use of PI3Kδ/γ inhibitor as an immune-regulatory radiosensitizer.

Long-term variation of nitrate in the East Sea, Korea.

Long-term variation of nitrate in the East Sea was monitored in order to investigate impact of Three Gorges Dam (TGD) in the Changjiang River's upstream, China and Nakdong River's estuary dam, Korea. Tracing source of nitrate was another objective in this study. For this study, nutrient data were collected for 20 years from 1999 to 2018 in the East Sea, and divided into 4 sections, and evaluated whether a significant difference exists among the averages of nitrate concentrations. The concentrations of nitrate were affected by the major rivers (the Nakdong and the Taehwa Rivers) and Tsushima Warm Current (TWC) which diverged from the Kuroshio Current passing through East China Sea (ECS). Our results also indicated that long-term nitrate concentrations decreased and its reasons. First, the construction of TGD in the upstream of the Changjiang River may have resulted in the decrease of the nitrate supply in the river and ECS which is carried by TWC, toward the East Sea. Second, decrease in the nitrate flux of the Nakdong River's estuary due to the construction of the estuary dam and sewer treatment plant could also be a factor for the nitrate decrease in the East Sea. Therefore, anthropogenic activities from the Nakdong River and Changjiang River had a long-term effect on the East Sea's nitrate concentrations. The amount of nitrate runoff reduced by the anthropogenic activities influenced the nitrate levels over a long period by the flow of currents in the East Sea.

Surgical Outcomes of Nonadjustable Modified Harada-Ito Surgery.

PURPOSE: This study aimed to investigate the surgical outcomes of nonadjustable Harada-Ito surgery under general anesthesia. METHODS: Twenty-two patients who underwent nonadjustable modified Harada-Ito surgery under general anesthesia were reviewed retrospectively. Among them, 21 out of the 22 patients who were followed up for 6 months after surgery were included in this study. Subjective cyclotorsion (double Maddox rod test) and objective cyclotorsions (fundus photography) were measured. Success of the surgery was defined as follows: success (the patients do not acknowledge diplopia at any direction), partial (the patients feel diplopia at a specific direction, but they do not feel discomfort in routine life), and fail (the patients feel diplopia in primary gaze, hence requiring a thorough investigation). RESULTS: The mean age of the patients (18 male and 3 female) was 56.5 years (range, 40-77 years). Based on the alternate prism cover test, the patients had 4.2 ± 3.0 prism diopters of vertical deviation. The corrected amounts of cyclotorsion based on the double Maddox rod test and fundus photography were 14.8° ± 7.5° and 9.8° ± 7.9°, respectively, and were significantly different between the two methods (p = 0.006). After the surgery, 20 out of the 21 patients (95.2%) completely recovered from diplopia in the primary gaze. However, among the 20 patients, seven complained of diplopia in the secondary gaze (down gaze, four patients; head tilt gaze, three patients). The success group had a smaller preoperative subjective excyclotorsion than the partial and fail groups (12.6° ± 2.5° and 21.0° ± 8.9°, respectively; p = 0.046). CONCLUSIONS: Nonadjustable modified Harada-Ito surgery under general anesthesia has favorable success rate, and preoperative subjective excyclotorsion can be a prognostic factor in patients with bilateral superior oblique palsy.

Discovery and Characterization of a Novel MASTL Inhibitor MKI-2 Targeting MASTL-PP2A in Breast Cancer Cells and Oocytes.

Although microtubule-associated serine/threonine kinase-like (MASTL) is a promising target for selective anticancer treatment, MASTL inhibitors with nano range potency and antitumor efficacy have not been reported. Here, we report a novel potent and selective MASTL inhibitor MASTL kinase inhibitor-2 (MKI-2) identified in silico through a drug discovery program. Our data showed that MKI-2 inhibited recombinant MASTL activity and cellular MASTL activity with IC50 values of 37.44 nM and 142.7 nM, respectively, in breast cancer cells. In addition, MKI-2 inhibited MASTL kinase rather than other AGC kinases, such as ROCK1, AKT1, PKACα, and p70S6K. Furthermore, MKI-2 exerted various antitumor activities by inducing mitotic catastrophe resulting from the modulation of the MASTL-PP2A axis in breast cancer cells. The MKI-2 treatment showed phenocopies with MASTL-null oocyte in mouse oocytes, which were used as a model to validate MKI-2 activity. Therefore, our study provided a new potent and selective MASTL inhibitor MKI-2 targeting the oncogenic MAST-PP2A axis in breast cancer cells.

Ablation of CRBN induces loss of type I collagen and SCH in mouse skin by fibroblast senescence via the p38 MAPK pathway.

Cereblon (CRBN) is a substrate receptor of the cullin-RING E3 ubiquitin ligase (CRL) complex that mediates the ubiquitination of several substrates. In this study, CRBN knockout (KO) mice exhibited decreased levels of stratum corneum hydration (SCH) and collagen I expression with an elevated protein level of matrix metalloprotease 1 (MMP1). The absence of cereblon in the skin of CRBN KO mice mimics the damage caused by narrowband ultraviolet B (NB-UVB). The primary CRBN deficient mouse embryonic fibroblasts (MEFs) undergo G2/M-arrested premature senescence via protein signaling of p38 MAPK and its dependent p53/p21pathway. The absence of CRBN induced the markers of cellular senescence, such as the senescence-associated heterochromatin foci (SAHF), SA-ß-Gal staining, and p21 upregulation while the ectopic expression of CRBN reversed the phenotypes of SA-ß-Gal staining and p21 upregulation. Reversion of the decreased protein level of collagen I was demonstrated after the reintroduction of the CRBN gene back into CRBN KO MEFs, validating the promising role of CRBN as a potential regulator for the function of the skin barrier and its cellular homeostasis.

MKI-1, a Novel Small-Molecule Inhibitor of MASTL, Exerts Antitumor and Radiosensitizer Activities Through PP2A Activation in Breast Cancer.

Although MASTL (microtubule-associated serine/threonine kinase-like) is an attractive target for anticancer treatment, MASTL inhibitors with antitumor activity have not yet been reported. In this study, we have presented a novel MASTL inhibitor, MKI-1, identified through in silico screening and in vitro analysis. Our data revealed that MKI-1 exerted antitumor and radiosensitizer activities in in vitro and in vivo models of breast cancer. The mechanism of action of MKI-1 occurred through an increase in PP2A activity, which subsequently decreased the c-Myc protein content in breast cancer cells. Moreover, the activity of MKI-1 in the regulation of MASTL-PP2A was validated in a mouse oocyte model. Our results have demonstrated a new small-molecule inhibitor of MASTL, MKI-1, which exerts antitumor and radiosensitizer activities through PP2A activation in breast cancer in vitro and in vivo.

SAR study of bisamides as cyclophilin a inhibitors for the development of host-targeting therapy for hepatitis C virus infection.

The therapy of chronic hepatitis C virus infections has significantly improved with the development of direct-acting antivirals (DAAs), which contain NS3/4A protease, NS5A, and NS5B polymerase inhibitors. However, mutations in specific residues in these viral target genes are associated with resistance to the DAAs. Especially inhibitors of NS3/4A protease and NS5A, such as grazoprevir and velpatasvir, have a low barrier to resistant mutations. As a result, the mutations influence the virological outcomes after DAA treatment. CypA inhibitors, as host-targeted agents, act on host factors to inhibit HCV replication, exhibiting a high resistance barrier and pan-genotype activities against HCV. Therefore, they can be developed into alternative, more effective anti-HCV agents. However, CypA inhibitors are natural products and analogs. Based on previous studies, bisamide derivatives were designed and synthesized to develop a novel class of CypA inhibitors. Bisamide derivative 7c is a promising compound with potent anti-HCV activity at subtoxic concentrations. Surface plasmon resonance experiments revealed that 7c directly binds to CypA. All these studies indicated that the derivative 7c is a potent CypA inhibitor, which can be used as a host-targeted agent in combination with other antiviral agents for anti-HCV treatment.

Barium Titanate Nanoparticles Sensitise Treatment-Resistant Breast Cancer Cells to the Antitumor Action of Tumour-Treating Fields.

Although tumour-treating fields (TTFields) is a promising physical treatment modality based on disruption of dipole alignments and generation of dielectrophoretic forces during cytokinesis, not much is known about TTFields-responsive sensitisers. Here, we report a novel TTFields-responsive sensitiser, barium titanate nanoparticles (BTNPs), which exhibit cytocompatibility, with non-cytotoxic effects on breast cancer cells. BTNPs are characterised by high dielectric constant values and ferroelectric properties. Notably, we found that BTNPs sensitised TTFields-resistant breast cancer cells in response to TTFields. In addition, BTNPs accumulated in the cytoplasm of cancer cells in response to TTFields. Further, we showed that TTFields combined with BTNPs exhibited antitumor activity by modulating several cancer-related pathways in general, and the cell cycle-related apoptosis pathway in particular. Therefore, our data suggest that BTNPs increase the antitumor action of TTFields by an electric field-responsive cytosolic accumulation, establishing BTNP as a TTFields-responsive sensitiser.

MRI Findings Predictive of Shoulder Stiffness in Patients With Full-Thickness Rotator Cuff Tears.

OBJECTIVE. The objective of our study was to evaluate the relationship between stiff shoulder in patients with a full-thickness rotator cuff tear and MRI findings, especially joint capsule abnormality. MATERIALS AND METHODS. This study included 106 patients with small to large (≤ 5 cm) full-thickness rotator cuff tears. Joint capsule edema and thickness in the axillary recess, obliteration of the subcoracoid fat triangle, fatty degeneration of the torn rotator cuff muscle, and degree of retraction were assessed by two radiologists. The size and location of tears were determined by MRI findings and operative report. Associations between MRI findings and preoperative passive range of motion (ROM) were assessed by simple and multiple linear regression analyses and proportional odds logistic regression analysis. RESULTS. There was a significant, negative linear correlation between limited ROM at forward elevation and thickness of the joint capsule in the glenoid portion of the axillary recess (p = 0.018), external rotation and joint capsule edema in the humeral portion of the axillary recess (p = 0.011), and internal rotation and joint capsule edema in the glenoid portion of the axillary recess (p = 0.007). Male sex (p = 0.041) and posterosuperior rotator cuff tear (p = 0.030) were independent predictors of shoulder ROM on external rotation. Degree of fatty degeneration (p = 0.003) was another independent predictor of shoulder ROM on internal rotation. CONCLUSION. MRI findings-especially joint capsule edema and thickness at the axillary recess-can be useful in predicting shoulder stiffness in patients with rotator cuff tear.

Antiviral activity of sertindole, raloxifene and ibutamoren against transcription and replication-competent Ebola virus-like particles.

A chemical library comprising 2,354 drug-like compounds was screened using a transcription and replication-competent viruslike particle (trVLP) system implementing the whole Ebola virus (EBOV) life cycle. Dose-dependent inhibition of Ebola trVLP replication was induced by 15 hit compounds, which primarily target different types of G protein-coupled receptors (GPCRs). Based on the chemical structure, the compounds were divided into three groups, diphenylmethane derivatives, promazine derivatives and chemicals with no conserved skeletons. The third group included sertindole, raloxifene, and ibutamoren showing prominent antiviral effects in cells. They downregulated the expression of viral proteins, including the VP40 matrix protein and the envelope glycoprotein. They also reduced the amount of EBOV-derived tetracistronic minigenome RNA incorporated into progeny trVLPs in the culture supernatant. Particularly, ibutamoren, which is a known agonist of growth hormone secretagogue receptor (GHSR), showed the most promising antiviral activity with a 50% effective concentration of 0.2 µM, a 50% cytotoxic concentration of 42.4 µM, and a selectivity index of 222.8. Here, we suggest a strategy for development of anti-EBOV therapeutics by adopting GHSR agonists as hit compounds. [BMB Reports 2020; 53(3): 166-171].

HNO2 treatment of sludge: An alternative way of sludge usage as fertilizer.

The research was conducted to evaluate the sludge quality as fertilizer after heavy metal removal by using free nitrous acid (FNA, HNO2) solution. To meet up the Korean criteria of fertilizer, FNA treatment with the ultrasonic process was studied here. The sample was taken from a local wastewater and sewage treatment plants and FNA with FNA plus ultrasonic used to treat them. As and Cd concentrations were met the Korean criteria after FNA treatment. In contrast, Al of sludge from sewage treatment plant did not decrease down to the criteria, possibly suggesting that it may be related to the coagulants of the sludge during thickening and dewatering. FNA treatment showed satisfactory results in comparison with the other acidic treatment of citric acid, oxalic acid, HNO3 or HCl. Removal efficiency of the Fenton-like reaction was more effective than the FNA method. Despite this result, the FNA treatment may be better for sludge recycling as fertilizer because the Fenton-like process removed the necessary organic matter, nitrogen as well as metals. Moreover, the addition of ultrasound process has in conserving time and expenditure in metal removal and FNA with ultrasound became more effective in shorter reaction time. Therefore, FNA treated sludge could be a better option for sludge recycling as fertilizer. In conclusion, a proper plan of sludge management should be required for future work to use sludge as fertilizer.

Predicting Radiation Resistance in Breast Cancer with Expression Status of Phosphorylated S6K1.

Emerging evidence suggests that the mammalian target of rapamcyin (mTOR) pathway is associated with radio-resistance in cancer treatment. We hypothesised that phosphorylated ribosomal S6 kinase 1 (p-S6K1), a major downstream regulator of the mTOR pathway, may play a role in predicting radio-resistance. Therefore, we evaluated the association of p-S6K1 expression with radio-resistance in breast cancer cell lines and patients. During median follow-up of 33 (range, 0.1-111) months for 1770 primary breast cancer patients who underwent surgery, patients expressing p-S6K1 showed worse 10-year loco-regional recurrence-free survival (LRFS) compared to that of p-S6K1-negative patients after radiotherapy (93.4% vs. 97.7%, p = 0.015). Multivariate analysis revealed p-S6K1 expression as a predictor of radio-resistance (hazard ratio 7.9, 95% confidence interval 1.1-58.5, p = 0.04). In vitro, CD44high/CD24low MCF7 cells with a radioresistant phenotype expressed higher levels of p-S6K1 than control MCF7 cells. Furthermore, the combination of radiation with treatment of everolimus, an mTOR-S6K1 pathway inhibitor, sensitised CD44high/CD24low MCF7 cells to a greater extent than MCF7 cells. This study provides in vivo and in vitro evidence for p-S6K1 expression status as an important marker for predicting the resistance to radiotherapy and as a possible target for radio-sensitization in breast cancer patients.

Valuation of Estimation Toxic Chemical Release Inventory Method-Focusing on Paint Manufacturing Process.

Industrial chemicals differ in their treatment methods and types, depending on their physicochemical properties. Highly volatile chemicals are emitted despite installation of preventive facilities, such as scrubbers and adsorption towers. Some countries release a Toxic Release Inventory (TRI), which is a mandatory report on the amount of chemicals emitted annually. This report is released to the citizens to ensure their right to knowledge and life. Numerous methods have been devised to investigate the amount of chemical emissions. There are four methods to estimate TRI emissions (Emission Factor Method; Material Balance Method; Source Testing Method; Emission Model Method). Moreover, efforts have been made to increase awareness and formulate plans to reduce chemical emissions. Despite this, the TRI method tends to underestimate and overestimate, especially due to volatile compounds. If the results of the TRI emissions are underestimated, toxic chemicals can have a negative impact on citizens. Volatile compounds are commonly used in chemical manufacturing plants, such as paint plants. In this study, a suitable method for each industrial process was suggested based on conservative estimates of multiple toxic chemical inventory method, focusing on the paint manufacturing process. In the paint manufacturing plant, storage, weighing, and mixing processes should be used emission model method to estimate TRI. In the reaction process, TRI must be estimated by the source test method. In the transfer process, the emission factor method should be used to estimate TRI. In the atmosphere prevention process, the emission factor method or source testing method should be used depending on the physical and chemical properties such as vapor pressure of the chemical.

MALDI-TOF MS-based total serum protein fingerprinting for liver cancer diagnosis.

Serum is one of the most commonly used samples in many studies to identify protein biomarkers to diagnose cancer. Although conventional enzyme-linked immunosorbent assay (ELISA) or liquid chromatography-mass spectrometry (LC-MS)-based methods have been applied as clinical tools for diagnosing cancer, there have been troublesome problems, such as inferior multiplexing capabilities, high development costs and long turnaround times, which are inappropriate for high-throughput analytical platforms. Here, we developed a simple and robust cancer diagnostic method using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based total serum protein fingerprinting. First, serum samples were simply diluted with distilled water and subsequently spotted onto a MALDI plate without prior chromatographic purification or separation. The sample preparation method was enough to collect reproducible total serum protein fingerprints and would be highly advantageous for high-throughput assay. Each of the integrated main spectrum profiles (MSPs), which are representative of liver cancer patients (n = 40) or healthy controls (n = 80), was automatically generated by the MALDI Biotyper 3 software. The reliability of the integrated MSPs was successfully evaluated in comparison with a blind test set (n = 31), which consisted of 13 liver cancer patients and 18 healthy controls. Additionally, our partial least squares discriminant analysis (PLS-DA) demonstrated a statistically significant difference in MALDI-TOF MS-based total serum protein fingerprints between liver cancer patients and healthy controls. Taken together, this work suggests that this method may be an effective high-throughput platform technology for various cancer diagnoses and disease evaluations.