Vibrio vulnificus infection. The combination of a TGC and ciprofloxacin has synergistic in vitro bactericidal activity against V. vulnificus. No clinical study has compared the standard regimen with TGC plus ciprofloxacin therapy for V. vulnificus infection.Combination therapy with a third-generation cephalosporin (TGC) and a tetracycline analogue is recommended for V. vulnificus infection at two medical centers in Korea from 1991 to 2016 were enrolled in this study. The patients were grouped according to the type of antibiotic administered. A retrospective propensity-score-matched case-control study of patients treated with TGC plus doxycycline or TGC plus ciprofloxacin was performed. The clinical characteristics and outcomes of the patients were analyzed.Patients with a confirmed V. vulnificus septicemia during the study, and the 30-day survival rate was 39% (85/218). The patients were classified into the following six treatment groups: TGC monotherapy (n = 82), TGC plus doxycycline therapy (n = 42), TGC plus ciprofloxacin therapy (n = 39), ciprofloxacin monotherapy (n = 14), other β-lactam monotherapy (n = 10), and other (n = 31). The survival rates of these groups were as follows: TGC monotherapy (35%), TGC plus doxycycline (38%), TGC plus ciprofloxacin (54%), ciprofloxacin monotherapy (29%), other β-lactam (20%), and other (39%). The 30-day survival rate showed no significant difference between the TGC plus doxycycline and TGC plus ciprofloxacin groups . Among the 81 patients treated with TGC plus doxycycline or TGC plus ciprofloxacin, 12 per treatment group were selected by propensity-score matching. There was no significant difference in the baseline characteristics or the frequency of fasciotomy between the two groups. The 30-day survival rate showed no significant difference between the TGC plus doxycycline (50%) and TGC plus ciprofloxacin (67%) groups .A total of 218 patients were confirmed to have V. vulnificus septicemia.Our data suggest that the outcome of TGC plus ciprofloxacin therapy was comparable to that of TGC plus doxycycline therapy in patients with  in vitro bactericidal activity against V. vulnificus. No clinical study has compared the standard regimen with TGC plus ciprofloxacin therapy for V. vulnificus infection. A total of 218 patients were enrolled who are confirmed to have V. vulnificus septicemia in two medical centers in Korea from 1991 to 2016. The 30-day survival rate was 39% (85/218) for all patients, 38% (16/42) for TGC plus doxycycline and 54% (21/39) for TGC plus ciprofloxacin . A propensity score-matched analysis was performed and 12 per treatment groups were selected. The 30-day survival rate showed no significant difference between the TGC plus doxycycline  and TGC plus ciprofloxacin  groups . The outcome of TGC plus ciprofloxacin therapy was comparable to that of TGC plus doxycycline therapy in patients with V. vulnificus septicemia.The combination of a third-generation cephalosporin (TGC) and ciprofloxacin has synergy  Vibrio vulnificus is a Gram-negative halophile that thrives in warm marine and estuarine environments worldwide [V. vulnificus produces toxins and enzymes, including capsular polysaccharides, metalloproteases, lipopolysaccharides, and cytolysin [V. vulnificus causes foodborne disease and wound infections. Primary septicemia caused by V. vulnificus is characterized by bacteremia without any obvious focus of infection and usually presents with a sudden onset of fever and chills, often accompanied by vomiting, diarrhea, abdominal pain, and pain in the extremities within 7 days after the ingestion of contaminated seafood. Within the first 24 h after the onset of illness, secondary cutaneous lesions such as cellulitis, ecchymosis proceeding to hemorrhagic bullae begin to appear on the extremities [V. vulnificus, with an average mortality rate exceeding 50% [V. vulnificus [orldwide . V. vulnytolysin , 3. Thesremities . This prding 50% –8. In adlnificus . Like sylnificus , 11.V. vulnificus, early surgical treatment (within 12–24 h of admission) is important to achieve a favorable outcome because the necrotic tissue has an insufficient blood supply to achieve an effective concentration of any antimicrobial agent [V. vulnificus in vivo or in vitro, including erythromycin, tetracycline, cephalosporin, tigecycline, ciprofloxacin and extended-spectrum penicillins [V. vulnificus infections with necrotizing skin lesions [V. vulnificus wound infections [V. vulnificus in a wound-infection model [V. vulnificus septicemia [V. vulnificus in vivo than previously used regimens in a model of subcutaneous wound infection [In patients with necrotizing skin and soft tissue infections (NSTI) caused by al agent . The rolicillins –17, clin lesions , 19. Ind lesions –22. Basefections . Fluoroqnfection .V. vulnificus infections [V. vulnificus. In addition, we evaluated prognostic factors for mortality in patients with a V. vulnificus infection.A study of mortality associated with all generation cephalosporin plus quinolone regimens, including 98 cases from The United States CDC Cholera and Other Vibrio Illness Surveillance (COVIS) dataset from 1990 to 2010 did not find sufficient clinical evidence of the efficacy of TGC plus ciprofloxacin against fections . We thusThe analysis of cases was approved by the institutional review board (IRB) of Chonnam National University Hospital and Chonnam National University Hwasun Hospital . All analysed data were anonymized.V. vulnificus infection and hospitalized at Chonnam National University Hospital  or Chonnam National University Hwasun Hospital  were enrolled. A systematic review of the patients’ medical records was performed. NSTI caused by V. vulnificus were diagnosed if both of the following conditions were met: (1) patients with infected skin lesions such as bullae, ecchymosis, or cellulitis and (2) the identification of V. vulnificus in blood and/or bulla or tissue culture specimens. Primary septicemia was defined as V. vulnificus bacteremia with no other obvious source of infection. Our hospitals serve Chonnam Province, including the surrounding islands , in South Korea. This region has a population of around 3.4 million persons.From January 1991 to December 2016, patients aged > 18 years diagnosed with a culture-confirmed Vibrio vulnificus isolates were identified by conventional methods using ID-GNB Vitek 2 cards . We collected clinical and laboratory information, including age, sex, pre-existing illnesses, site of the NSTI, involved area, symptoms at admission, vital signs, physical and laboratory findings at the time of admission, type of surgical treatment, timing of surgery, antibiotics used, and outcomes. The primary outcomes were the 30-day mortality rate. The severity of illness at admission was evaluated using the first-day Acute Physiology and Chronic Health Evaluation (APACHE) II score. Sepsis and septic shock were defined according to the guidelines of the American College of Chest Physicians and Society for Critical Care Medicine [Medicine . EnrolleMedicine .t-test or the Mann-Whitney U test for continuous variables.Categorical variables were expressed as percentages and continuous variables as means and standard deviations (SD) or medians and interquartile ranges (IQR). In univariate analysis, Pearson’s chi-squared or Fisher’s exact test was used for comparisons of dichotomous variables, and Student’s P < 0.20) in the univariate analysis was performed; a value of P < 0.05 was regarded as indicative of statistical significance. The examined variables included infection-related factors that had the potential to modulate mortality, in addition to other demographic and therapeutic variables.Variables of the fatal and nonfatal groups were compared by univariate analysis to identify risk factors for mortality. Next, a multiple logistic regression analysis of factors found to be significant (V. vulnificus infection) [A Kaplan-Meier survival analysis and the log-rank test were performed to compare the efficacy of the antibiotic regimens. Propensity-score adjustment was performed to adjust for the following confounding variables: chronic hepatitis B, white blood cell count, shock at admission, early surgical intervention, APACHE II score, heavy alcohol drinking, hemoglobin level, platelet level, and the involvement of two or more extremities (which is associated with the prognosis of fection) , 28, 29.V. vulnificus septicemia were enrolled in the study. Among them, V. vulnificus was isolated from the blood of 171 patients, the tissue of 68 patients, and the bullae of 56 patients. Overall, 199 patients  had necrotizing skin lesions and 81 patients  died within 48 h of arrival; the 30-day mortality rate was 61% (133/218). The demographic, clinical, and laboratory characteristics of the patients are summarized in In total, 218 patients with During the study period, various antibiotics were administered. The patients were classified into six treatment groups according to the antibiotic(s) administered . In total, 82 patients (38%) received TGC monotherapy, 42 patients (19%) underwent TGC plus doxycycline therapy, 39 patients (18%) received TGC plus ciprofloxacin therapy, and 14 patients (6%) underwent ciprofloxacin monotherapy.P = 0.22). Pairwise post hoc analysis showed statistical significance only between TGC plus ciprofloxacin and other β-lactam (P = 0.02).The survival rates at 30 days after hospital admission are presented in P = 0.06) or ciprofloxacin monotherapy group (P = 0.057). However, there was no significant difference in survival rate between the TGC plus ciprofloxacin group and TGC plus doxycycline groups (P = 0.18). There was no significant difference in survival rate between the TGC monotherapy and TGC plus doxycycline groups (P = 0.76).A Kaplan-Meier survival analysis and log-rank test  showed tThe demographic characteristics of the TGC plus ciprofloxacin and TGC plus doxycycline groups are shown in V. vulnificus septicemic patients who had not received any surgical treatment  (The 24 patients in the two groups were 1:1 propensity-score matched . In the  = 0.46) .i.e., the 30-day mortality increased by 24% for each increment in the APACHE II score).In the multivariate analysis, septic shock at admission and an elevated APACHE II score were independent risk factors for mortality, and early surgical intervention was an independent prognostic factor for a lower mortality rate . The adjV. vulnificus. Of the V. vulnificus-infected patients, 75% received TGC-based antibiotics. The TGC plus ciprofloxacin group had the highest 30-day survival rate (54%). In the propensity score-matched study, the 30-day survival rate of the TGC plus doxycycline group was 6/12 (50%) and that of the TGC plus ciprofloxacin group was 8/12 (67%). Therefore, TGC plus ciprofloxacin was as effective as TGC plus doxycycline. Early surgical intervention (within 24 h of admission) was an independent prognostic factor for survival.In this study, > 90% of patients had necrotizing skin lesions. The early mortality rate was 37% and the 30-day mortality rate was 61% in patients with septicemia and/or NSTI caused by vs. 55%) and death (61% vs. 17%) than those with wound infections [According to US surveillance data, patients with foodborne illness have higher rates of septicemia  and < 30% of the patients had hepatic disorders. Hepatic diseases could be poor prognostic factors because the elevated serum ferritin level associated with these conditions promotes the survival of V. vulnificus in whole blood [V. vulnificus in seafood, including oysters, purchased from fish markets in Korea reported rates of resistance to cefotaxime, tetracycline, and ciprofloxacin of 11.8%, 5.9%, and 17.6%, respectively [V. vulnificus has been confirmed in several regions [V. vulnificus isolates showed that 45% of the environmental isolates were resistant to three or more classes of antibiotics and 17.3% were resistant to eight or more antibiotic agents [V. vulnificus infections .TGC plus doxycycline combinations are standard treatments for  America , 32. Chea [et al  reportedle blood . In thisectively . The emeV. vulnificus septicemia, the survival rate of the ceftriaxone plus ciprofloxacin group was 100%, compared with 91% for the ceftriaxone-doxycycline group; the difference was not significant [V. vulnificus in vitro and in vivo [V. vulnificus is mediated by inhibition of the transcription of repeats-in-toxin [In a mouse model of foodborne nificant . In thisnificant , 19. Con in vivo , 37. The in vivo . The addin-toxin . Ciproflin-toxin –41. The V. vulnificus septicemic patients of any single-institution study. The main limitation was that this was a 26-year retrospective analysis. Even though there was no significant change of outcome between the first and the second half of the time , the long time period could make potential for multiple confounding factors. TGC plus ciprofloxacin therapy was frequently administered in more recent years. Very diverse antibiotics in the six treatment groups were administered and individual decisions about antibiotics selection may result in selection bias. Although the internist communicated with the surgeon regarding the need for surgical intervention as soon as possible after the patient’s arrival at the emergency room, the decision to perform debridement (as well as the timing thereof) was not made based on standardized criteria. Many patients did not receive early surgical intervention due to various reasons such as host factors and reluctance of family to surgery, etc. The small portion of patients with early surgical intervention may be a confounding factor. Nevertheless, we believe that this is the actuality of the real world. We recommend temporizing surgical management according to the spread of skin lesions in patients with foodborne primary V. vulnificus septicemia with multi-extremity involvement because such patients are not clinically homogeneous. Second, various TGCs were administered and oral doxycycline was available only. Third, we could not evaluate the clinical efficacy of ciprofloxacin monotherapy due to the high early mortality rate of the patients treated with this agent. Lastly, we did not obtain a sufficient sample size to ensure adequate power to show difference. For two-sample noninferiority log-rank test by the statistical power 0.8, each 60 sample size are needed. Further research is thus warranted.This work included the largest number of V. vulnificus infections. Our propensity-score-matched analysis showed no significant difference in efficacy between TGC plus ciprofloxacin and TGC plus doxycycline regimens. This is to our knowledge the first clinical report of the non-inferior efficacy of TGC plus ciprofloxacin compared with TGC plus doxycycline in patients with V. vulnificus septicemia. The results suggest that TGC plus ciprofloxacin should also be a CDC recommended treatment for patients with a V. vulnificus septicemia.Their sporadic occurrence hampers determination of the efficacy of antimicrobials against human


Ovarian cancer is the fifth leading cause of cancer-related female mortality and the most lethal gynecological cancer. In this report, we present a rare case of recurrent granulosa cell tumor (GCT) of the ovary. We describe the case of a 26-yr-old woman with progressive GCT of the right ovary despite multiple lines of therapy who underwent salvage therapy selection based on a novel bioinformatical decision support tool (Oncobox). This analysis generated a list of potentially actionable compounds, which when used clinically lead to partial response and later long-term stabilization of the patient's disease. Ovarian cancer is the fifth most common cause of cancer-related death among women and the most lethal gynecological malignancy . WorldwiGranulosa cell tumor (GCT) of the ovary constitutes 2%–5% of all ovarian malignancies . Most caSurgical treatment  is also recommended by NCCN for treatment of GCT relapses and metastases. Common metastatic lesions include neoplasms in both the pelvic area and the parenchymal organs.Adjuvant platinum-based chemotherapy is recommended for patients with a high risk of recurrence. In the presence of residual neoplasms, regimens that include platinum drugs are effective in a considerable proportion of cases (∼60%) . In thisA 26-yr-old woman was diagnosed at N.N. Blokhin Russian Cancer Research Center with primary GCT of the right ovary in 2001. The patient underwent unilateral salpingo-oophrectomy, with peritoneal biopsies without evidence of tumor growth in the left ovary and solitary complexes of malignant cells. From 2003 until 2008, the patient underwent three excisions of the following neoplasms: cystadenoma in the left ovary , cystic formation in the left ovary, and GCT in the right lateral region of the abdominal cavity.Dissemination of neoplastic foci on the patient's peritoneum was observed in 2010. Ultrasound examination revealed a 2.9 × 0.8-cm neoplasm in the S7 liver capsule; the dimensions of cystic formation in the pelvis were 4.0 × 3.2 × 2.6 cm. The patient received megestrol  for 5 mo; however, lesions progressed during this period. Extirpation of recurrent tumors in the pelvic area was performed with dissection of adhesions and resection of the large omentum.Supplemental Table S1. The largest formations were identified on the posterior surface of the liver  and in the right part of the posterior paranephric fat . A 4.8 × 4.5-cm neoplasm in the Douglas space (neoplasm #2) displaced the rectum to the right. The patient underwent cytoreductive (debulking) surgery. Hematoxylin and eosin staining confirmed the primary origin of the tumor  using immunohistochemistry. The sample appeared to be c-Kit-negative, in accordance with microarray data . The patient was administered sorafenib  from October 2015. The expression level of sorafenib target genes is presented in To identify further treatment options, we performed molecular analysis of the patient's tumor. We extracted DNA from the patients’ formalin-fixed, paraffin-embedded (FFPE) tumor tissue sample, obtained following cytoreductive (debulking) surgery in 2013, and performed whole-exome sequencing. The sequencing data were deposited in the NCBI Sequence Read Archive (SRA) under accession ID PRJNA503667. The tumor appeared to be FOXL2 C134W-positive, which corresponds to the adult-type GCT of the ovary . We alsoSupplemental Fig. S1); the formation in the Douglas space in the pelvis decreased in size to 12.7 × 8.7 cm  was administered from February until May 2016. An MRI examination in March 2016 revealed a slight increase in sizes of the neoplasms. Neoplasm #1, located in the capsule of the right lobe of the liver, was 12.0 × 10.4 × 10.2 cm in size ; in the Supplemental Table S1; Supplemental Table S1; From May until July 2016, imatinib was replaced by the filachromine (generic Imatinib) because of the unavailability of original Imatinib (Gleevec). MRI examination in June indicated stabilization of the disease (MRI of the abdomen and pelvic area performed in December 2016 (source images were not available) revealed that the cystic-solid nodules under the right lobe of the liver (13.5 × 12.0 cm) squeeze the liver and liver’s gates; the nodules in the right lateral canal (7.8 × 5.5 cm) displaced the right kidney upward and medially. The dimensions of the tumor in the pelvic cavity were 17.0 × 11.0 cm, the rectum was shifted to the right and compressed, and the bladder was compressed and shifted anteriorly.Another cytoreductive surgery was performed in December 2016. Neoplasms in the right lobe of the liver (#1) and in the pelvic area (#5) were partially removed. Revision of the abdominal cavity revealed significant adhesive process. Tumor nodules with a thin capsule were intimately connected with the bladder, rectum, and ureters. Operative blood loss was 4 L. The metastatic nodules were removed in part because of technical difficulties. The ultrasound examination performed after surgery revealed that the liver was enlarged with the right lobe pushed back and pressed by a cystic neoplasm . Similar neoplasm of size 8.4 × 6.6 cm (neoplasm #3) was located laterally.Supplemental Table S1). A new cystic neoplasm  was observed during MRI examination in October. The sum of the largest diameters for all target lesions increased by 12% when compared to the MRI results from September 2016 (considering only the lesions investigated during both examinations), which supports disease stabilization.Original imatinib (Gleevec) became available and its administration (400 mg daily) started in February 2017. Before imatinib treatment, ultrasound examination in February 2017 revealed a tumor in the region of the posterolateral liver capsule , in the splenic hilum , and in the area of the lateral channel in the abdominal cavity  and several neoplasms in the pelvic region . MRI analysis in October 2017 revealed a decrease in size for these lesions, with the exception of the neoplasm in the splenic hilum area (its size was stable) , which corresponds to disease stabilization. As of June 2018, the patient is alive and physically active with a Karnofsky scale index of 90%. Imatinib administration is continued, no significant side effects are observed, and no surgical procedures are required.An MRI examination in March 2018 revealed moderate growth of lesions #3, #4, #10, and #13 (Tyrosine kinase inhibitors (TKIs) represent a class of target drugs that have been widely integrated into clinical practice since the beginning of the twenty-first century . ProteinKit-positive unresectable malignant gastrointestinal stromal tumors , KIT, and several other TKs . Imatinil tumors . The Phal tumors . Severall tumors ; howeverl tumors .Here, we report a case of adult-type recurrent ovarian GCT, successfully treated with imatinib monotherapy. Although the disease progressed during best supportive care, imatinib treatment resulted in a prolonged stabilization of the disease.Importantly, the prescription of imatinib was based on the bioinformatical analysis of gene expression data from the patient's tumor biopsy (Oncobox platform). Moreover, sorafenib treatment, which was also suggested by Oncobox, resulted in a partial tumor response and was terminated only because of significant side effects.We conclude that a personalized approach for TKI prescription in ovarian cancer is needed. The selection of patients who may potentially benefit from imatinib or other TKI treatment may be based on the molecular profiling of tumor biopsies and further bioinformatical analysis. However, further extended clinical trials are required for validation and adjustment of clinical indications for this particular bioinformatical platform Oncobox.Supplemental Table S4. Mutation calling was performed using the Picard and Genome Analysis Toolkit.An FFPE block with >80% of tumor cells was analyzed. We extracted RNA from five 250-µm-thick simultaneously made sections of this block. DNA was extracted from the FFPE tissue using the AnaPrep FFPE DNA extraction kit following the manufacturer's instruction. Whole-exome DNA was captured from total genomic DNA using the SeqCap EZ System from NimbleGen according to the manufacturer's instructions. Briefly, genomic DNA was sheared and size selected to roughly 200–250 base pairs and the ends were repaired and ligated to specific adapters and multiplexing indexes. Fragments were then incubated with SeqCap biotinylated DNA baits followed by the ligation-mediated PCR, and the RNA–DNA hybrids were purified using streptavidin-coated magnetic beads. The RNA baits were then digested to release the targeted DNA fragments, followed by a brief amplification of 15 or fewer PCR cycles. Sequencing was performed using Illumina HiSeq 3000. The reads were aligned with BWA-MEM. Sequencing coverage table is available in Inc (USA). The Manufacturer's protocol was modified by adding to the amplification reaction dNTP mix containing biotinylated dUTP, resulting in a final proportion of dTTP/biotin-dUTP as 5/1. Hybridization was performed according to CustomArray ElectraSense Hybridization and Detection protocol. The hybridization mix contained 2.5 µg of labeled DNA library, 6× SSPE, 0.05% Tween 20, 20 mM EDTA, 5× Denhardt's solution, 100 ng/µL sonicated calf thymus gDNA, and 0.05% SDS. The hybridization mix was incubated with chip overnight at 50°C. Hybridization efficiency was detected electrochemically using the CustomArray ElectraSense Detection Kit and ElectraSense 4X2K/12K Reader.The gene expression profile in the mixed sample was analyzed using the microarray platform CustomArrayGene expressions of more than 3000 human genes were profiled and deposited at GES under accession ID GSE112579. The analysis of gene expression peculiarities was performed based on comparison with four samples of healthy ovarian tissues . Gene expression profiles were pooled and quantile-normalized using the R statistical programming language and “preprocessCore” library.d is the target drug under investigation; a and b are the weight coefficients varying from −1 to 1.5 depending on the target drug type; the drug efficiency index for molecular pathways DESMPd is calculated based on the activity levels for molecular pathways containing molecular targets of drug d; and the drug efficiency score for target genes DESTGd is calculated based on levels of expression of individual gene products.The profiling of intracellular signaling pathways altered in the patient's tumor tissue when compared to normal was performed using the Oncobox bioinformatical platform. The Oncobox system is capable of modeling the drug's ability to block pathological changes in molecular pathways and simultaneously block gene products with a pathological increase in the expression level. In contrast to other known analogs, the Oncobox platform uses the parameter of the balanced efficiency score (BES) for each drug as a target drug efficiency measure. Wherein, the data on molecular pathway activity in a test sample and the data on expression levels of gene products—targets of a certain drug—are simultaneously used for the BES calculation. The BES value is calculated according to the formulaMP, the formulad is the unique identifier of the target drug; t is the unique identifier of the gene product, the target of drug d; p is the unique identifier of the signaling pathway; PALp is the molecular pathway p activation strength; and the discrete value AMCF (activation-to-mitosis conversion factor) is to be determined as follows:To calculate DESTG, used is the unique identifier of the target drug; t is the unique identifier of the gene product, molecular target of drug d; p is the unique identifier of the signaling pathway; CNRt  is the ratio of the expression levels of the protein-coding gene t in the test sample to the norm (averaged expression level for a control group); ln is the natural logarithm; the definitions of DTIdt,, AMCFp, and NII are similar to those given above. The discrete value ARRtp (activator/repressor role) is defined for a gene product t in the pathway p as follows and deposited into the molecular pathway database:The discrete value DTI (drug–target index) is defined asd, weight coefficients a and b are used, which differ depending on the drug type.To calculate the BES for drug a = 0 and b = 1, because these drugs directly block the VEGF molecules in the blood flow while not binding with the molecular targets inside the cell or on the cell surface and, therefore, do not directly affect the intracellular signaling. For monoclonal antibodies that bind with their molecular targets on the cell surface (mAbs), a = 0 and b = 1, as their main mode of action consists in activation of immune cytotoxic response against the cells having bound mAb molecules on their surface and does not deal with strong modulation of signaling by affecting molecular pathways. Killer mAbs consist of antibodies against molecular targets on the cell surface chemically bound with cytotoxic agents. When binding with their targets on the cell surface, the killer mAbs kill these cells, thus demonstrating therapeutic mechanism not related to intracellular molecular pathway activation. For them, a = 0 and b = 1.5; in this case, the increased coefficient b represents proprietary high cytotoxic activities of these drugs. For drugs blocking de novo tubulin polymerization, a = 0 and b = 1; this represents the indefinite function of many targeted pathways for these drugs in cell survival and proliferation, as well as their direct inhibitory effect on their molecular targets. The same coefficients are also set for histone deacetylase inhibitors for the same reasons concerning their mechanism of action. For DNA-alkylating agents, a = 0 and b = −1, reflecting the indefinite functions of the majority of targeted pathways for cell survival and proliferation, as well as the direct inhibitory effect of these drugs on DNA repair proteins that target the alkylated DNA (reflected by the coefficient b = −1). For immunotherapeutic drugs, both coefficients are equal to 0.5, because of the dependence of their effect on the availability of both direct molecular targets and molecular pathway activation profiles related to tumor infiltration with lymphocytes. Similarly, the poly-ADP ribose polymerase blocking drugs inhibit DNA repair and depend on both availability of direct molecular targets and on the activities of targeted molecular pathways. This is reflected by both coefficients a and b being equal to 0.5.For low-molecular TKIs (nibs), both weight coefficients are equal to 0.5, representing the equal significance of the target molecular pathway activation and target gene expression levels in the pathological tissue sample tested. This is related to the nibs’ capability of blocking their molecular targets and thus inhibiting their activities, as well as modulating the cell signaling via related molecular pathways. For hormones, both weight coefficients are equal to −0.5, because they activate but do not inhibit their molecular targets and act accordingly also on their target molecular pathways. For antihormones, coefficients are equal to 0.5 again, which is because of their inhibition effect on their molecular targets, hormone products, and on the respective molecular pathways. For retinoids, both coefficients are equal to 0.5 because these drugs bind retinoic acid receptors and activate a number of dependent molecular pathways. For rapalogs , both coefficients are equal to 0.5 because they demonstrate their inhibition effect by directly binding with their molecular targets and act accordingly on the relevant molecular pathways. For mibs (proteasome inhibitors), both coefficients are equal to 0.5 because these drugs demonstrate the inhibition effect when binding with their molecular targets and act accordingly on the relevant molecular pathways and proteasome signaling. For VEGF blocking agents, https://www.ncbi.nlm.nih.gov/geo/) under accession number GSE112579. Whole-exome sequencing data were deposited to NCBI SRA under accession number PRJNA503667.Gene expression data derived from the patient's tumor tissue were deposited at the GEO  at Clinical Center Vitamed, Moscow, Russia, according to the principles of the Declaration of Helsinki.E.V.P., M.P.B., D.O.A., and V.B. collected and interpreted patient data. E.V.P. and M.P.B. were involved in clinical management. M.V.S. and D.E.K. performed molecular analyses. L.M. analyzed and interpreted the MRI data. M.V.S., A.A.A., I.N.K., and E.V.P. wrote this manuscript.The authors have declared no competing interest.


D) were disclosed, such as with the height of the Taylor cone , with the angle of the Taylor cone , with the length of the straight fluid jet , and with the spreading angle of the instable region . In vitro dissolution tests verified that the smaller the diameters, the faster ketoprofen (KET) was released from the HPMC nanofibers. These concrete process-property relationships should provide a way to achieve new knowledge about the electrostatic energy-fluid interactions, and to meanwhile improve researchers’ capability to optimize the coaxial process conditions to achieve the desired nanoproducts.The concrete relationship between the process parameters and nanoproduct properties is an important challenge for applying nanotechnology to produce functional nanomaterials. In this study, the relationships between series of process parameters and the medicated nanofibers’ diameter were investigated. With an electrospinnable solution of hydroxypropyl methylcellulose (HPMC) and ketoprofen as the core fluid, four kinds of nanofibers were prepared with ethanol as a sheath fluid and under the variable applied voltages. Based on these nanofibers, a series of relationships between the process parameters and the nanofibers’ diameters ( Nanofibers, as a special kind of nanomaterial, have the unique physical properties of an individual nanoscale diameter assembled into a macroscale film, which endow them with the capability to connect the invisible nano world with the visible macro world ,2,3,4. CH and α), the straight fluid jet , and also the bending and whipping region  [The diameter of nanofibers has a close relationship with their functional performance, which has prompted researchers to try their best to reduce their diameter ,39. Basengle, θ) .Electrospinning was revived from 1995 and was once regarded as a successive splitting process within the atomization region for creating nanofibers . Almost Previously, a series of mathematical models have been put forward in the literature, which focus on the relationship between the experimental parameters and the final nanofibers’ quality ,49,50. TCoaxial electrospinning is an upgraded version of the conventional blending electrospinning process, in which a concentrate spinneret is explored to lead two different kinds of working fluids into the electric field in a core-sheath manner ,52. TradBased on the above-mentioned knowledge, here for the first time, we investigated the relationships between the process parameters and the electrospun polymeric nanofibers’ diameter. With hydroxypropyl methylcellulose (HPMC) as a filament-forming polymeric matrix ,57, seveKET was provided by Wuhan Anruike Biological Pharmaceutical Co., Ltd. . HPMC powder  was obtained from Shandong Fine Chemical Co., Ltd. . Ethanol and dichloromethane (DCM) were purchased from Shanghai Lingfeng Chemical Testing Co. Ltd. . All other chemicals are analytical reagents, and water was distilled twice before use.An electrospinnable solution consisting of 13% (w/v) HPMC and 3% (w/v) KET in a mixture of ethanol and DCM  was prepared and utilized as the core fluid. Pure solvent ethanol was used as the shell fluid. Four nanofibers referred to as F1, F2, F3, and F4 were prepared at applied voltages of 13, 14, 15, and 16 kV, respectively. For all preparations, the fiber-collected distance was fixed at 12 cm. The shell and core fluid flow rates were 0.2 and 0.8 mL/h, respectively. The modified coaxial processes were recorded using a digital camera .The surface morphological characterization of the prepared nanofibers was observed using scanning electron microscopy  at 30 kV of accelerated voltage. Before the observation, the samples were sputter-coated with gold in a vacuum. The images were analyzed by ImageJ software with over 100 different places measured.20 mg of nanofibers was placed in 100 mL of phosphate buffer solution (PBS) with a pH value of 7.0. The buffer solution including samples was incubated in a shaking bath at 37 ± 0.1 °C and an agitation speed of 50 rpm. This was repeated 6 times for each kind of sample. At the predetermined time intervals, 1 mL of sample solution was withdrawn and replaced with 1 mL fresh medium. The amount of KET released from the nanofibers was measured using a UV-Vis Spectrophotometer by measuring the absorbance at 260 nm. The calibration curve was obtained, and all concentrations were evaluated in percentage as mean ± standard deviation using the following Equation (1).Modified coaxial electrospinning, derived from the traditional coaxial process, has several advantages over the latter. It not only can be similarly utilized to created core-shell nanostructures , but alsThe modified coaxial electrospinning can greatly expand the capability of electrospinning in generating new nanostructures ,49. ThisThe implementation of modified coaxial electrospinning is shown in H) gradually decreased, whereas the angle (α) gradually increased as the applied voltage gradually increased. Estimated by the outer diameter of Teflon tubing of 3 cm, the height of Taylor cones were 2.08 ± 0.32, 1.61 ± 0.27, 1.35 ± 0.26 and 1.15 ± 0.24 cm and the cone angles were 34 ± 6°, 38 ± 7°, 40 ± 7° and 42 ± 6° as the voltages elevated from 13 to 14, 15, and 16 kV, respectively (n = 3).L) straight fluid jet decreased from 2.87 ± 0.34, to 2.04 ± 0.27, 1.57 ± 0.31 and 1.23 ± 0.24 cm, whereas the spreading angle of bending and whipping increased from 39 ± 5°, to 45 ± 6°, 51 ± 5°, and 56 ± 8°, respectively (n = 3). Although the applied voltage has a certain electrospinnable range, within which electrospun HPMC nanofibers can be similarly created, the working fluids were highly sensitive to the changes in voltage. Moreover, because the modified coaxial electrospinning process started from the nozzle of Teflon tubing, the images of the Taylor cone, straight fluid jet and instable region could be captured simultaneously with Teflon tubing. Thus, the outer diameter could be utilized as a scale to estimate the height of Taylor cone, the cone angle, and the length of the straight jets.The alternations of the straight fluid jet and the instable region are exhibited in The SEM images of the prepared KET-loaded HPMC nanofibers and their diameter distributions are given in D = 2190 − 104V, with a correlation coefficient VR2= 0.9736. The applied voltage is an operational parameter that can be manipulated directly by the researchers, and thus is frequently utilized to downsize the prepared nanofibers [Shown in nofibers ,40,41,42nofibers . The modnofibers ,70,71,72D) with the height of the Taylor cone (H), whose linear equation is D = 125 + 363H, with a correlation coefficient hR2= 0.9884. D) with the angle of the Taylor cone (α), whose linear equation is D = 1576 − 19α, with a correlation coefficient Rα2= 0.9865. These results suggested that the applied voltage influenced both the Taylor cone’s height and angle, and the Taylor cone’s volume.The change in applied voltage can result in a successive response from the electropspinning working process to the final nanoproducts. Thus, the process parameters that can be utilized to characterize the working process should have a certain relationship with the nanofibers’ diameter. To fit the height and angle of the Taylor cone with the fiber’s diameter, two fine linear relationships can be found for them. Shown in D) and the length of the straight fluid jet (L), whose equation is D = 285 + 209L, with a correlation coefficient lR2= 0.9912. The higher linear correlation suggests that the longer the length of straight fluid jet, the larger diameter the nanofibers have. This suggests that the length of the straight fluid jet can be a useful process parameter for accurately predicting the diameter of the resulting nanofibers.Shown in D) with the spreading angle of the instable region (θ), whose equation is D = 2342 − 43θ, with a correlation coefficient Rθ2= 0.9857. The larger the spreading angle is, the smaller the final nanofibers. This fine linear correlation suggests that the spreading angle of the instable region can also be a useful process parameter for accurately predicting the resultant nanofibers’ diameter.Shown in The equations in In practice, any changes in the experimental conditions  will equally exert their influences on both the working processes and the successive solid nanofibers. Thus, the successful monitoring of the working process and accurate measurement of the process parameters would greatly increase our capability of accurate prediction about the size of final products and our capability to perform elaborate manipulation of the electrosprinning processes. As far as the measurement of process parameters is concerned, a high frequency camera can be exploited to record the working processes and to provide more accurate process parameter values than this work.It is well known that the prerequisite for the initiation of electrospinning is enough electrical force resulting from a large applied voltage, which is exploited to overcome the surface tension of the droplets pumped from the nozzle of spinneret. After a critical voltage value, the elevation of applied voltage would still revise the elecrospinning process, and often the higher the applied voltage applied, the smaller the Taylor cone, the shorter the straight fluid jet, and the finer the nanofibers created. In the traditional single-fluid electrospinning, the semi-vertical angle of the Taylor cone (α), representing the sharpness of the liquid hyperboloid, is often between 32° to 46° . In the G, which can often be neglected), the repelling forces between the up-down bending loops (F2), and the repelling forces between the adjacent places within the fluids (F3). It is the force F3 that mainly takes charge of the size reducing effect during electrospinning. During this process, the spreading angle is a key process parameter that reflects the synergetic actions of F1, F2 and F3. The elevation of applied voltage would increase all three forces. The increase in force F3 would expand the bending and whipping loops and thus increase the spreading angle θ, and promote the fluid drawn effects. The increase of F2 would make the fluid move slowly for more time during the drawing process, which similarly helps to enhance the trend of enlarging the spreading angle. The F1 should accelerate the working fluids flying to the collector and reduce the spreading angle. However, the combined effect of F2 and F3 is greater than F1, and thus the apparent result is that the bigger the applied voltage, the larger value the spreading angle has.During the electrospinning process, there are often several kinds of forces exerted on the working fluids. Shown in Shown in A-A to B-B). In this process the water molecules penetrate the solid nanofibers. The solid nanofibers gradually become a transparent hydrogel. Meanwhile, the drug molecules should leave the polymer chains and go into the penetrated water, and further diffuse outward to the bulk solution due to the concentration gradient. This is a drug diffusion mechanism. As the swelling goes forward, the outer layer HMPC molecules will free themselves into the bulk solution, together with the contained and penetrated drug molecules, until complete disappearance (from B-B to C-C and D-D). Thus, the erosion mechanism also happens here.HPMC is a soluble and hydrophilic polymer. Shown in As the nanofiber diameters decrease, the penetration distance of water and diffusion distance of drug molecules should all decrease correspondingly. This is to say the decrease of diameter will shorten the drug release time period due to the diffusion mechanism. This should be the reasons that the smaller the nanofibers’ diameter, the more positive the results for the fast dissolution of the poorly water-soluble drug. The poor water-solubility of drugs is one of the most difficult and long-existing issues in pharmaceutics ,75,76,77D) was estimated and a series of relationships between it and the process parameters were disclosed, such as with the height of the Taylor cone , with the angle of the Taylor cone , with the length of the straight fluid jet , and also with the spreading angle of the instable region . All these relationships show a slightly better linear relationship than the applied voltage experimental parameter , as indicated by their correlation coefficients. In vitro dissolution tests verified that the smaller the diameters, the faster the release of KET from the HPMC nanofibers. These clear process-property relationships should provide a way to achieve new knowledge about the electrostatic energy-fluid interaction process, and meanwhile improve the capability of researchers in optimizing coaxial process conditions to achieve the desired nanoproducts.In this study, modified coaxial electrospinning was successfully carried out using a Teflon-coated concentric spinneret. Four types of KET-loaded HPMC nanofibers were created under a variable experimental parameter—applied voltage. Based on the SEM images, the nanofibers’ diameter (


We aimed to evaluate the accuracy of coronary computed tomography angiography (CCTA) with a low-radiation protocol and iterative model reconstruction (IMR), in comparison with invasive coronary angiography (ICA). Sixty-one patients  with suspected coronary artery disease who underwent CCTA and ICA were retrospectively enrolled. CCTA was performed with low tube voltage (80 or 100 kVp), low tube current (100–200 mAs), prospective ECG triggering, and IMR using a 64-slice computed tomography scanner. Coronary artery disease was defined as luminal narrowing of >50%, as assessed using CCTA and ICA. The sensitivity, specificity, positive (PPV) and negative (NPV) predictive value, and accuracy of CCTA were examined. The mean radiation dose of CCTA was 1.05 ± 0.36 mSv. No non-diagnostic segment was noted. The sensitivity, specificity, PPV, NPV, and accuracy of CCTA were 86.4%, 96.1%, 80.3%, 97.5%, and 94.6% on a per segment basis, 93.1%, 94.7%, 88.3%, 97.0%, and 94.2% on a per vessel basis, and 100%, 83.3%, 93.5%, 100%, and 95.1% on a per patient basis, respectively. In conclusion, a low-radiation CCTA protocol with IMR may be useful for diagnosing coronary artery disease, as it reduces the radiation dose while maintaining diagnostic accuracy. The diagnostic performance of prospectively ECG-triggered CCTA with lowered radiation exposure by decreasing the scanning duration showed a high negative predictive value (NPV), when using stenosis by invasive coronary angiography (ICA) as the reference in patients with low-to-intermediate probability for CAD6.Coronary computed tomographic angiography (CCTA) is a useful, non-invasive diagnostic imaging modality for the detection of coronary artery disease (CAD). Various strategies and scanning protocols enable a reduction in the radiation dose used for CCTA, including high-pitch spiral acquisition, prospective ECG triggering, tube voltage lowering, and tube current modulation8. To our knowledge, a few recent studies have evaluated the diagnostic performance and clinical feasibility of CCTA for CAD using low tube voltage and the model-based iterative reconstruction algorithm (MBIR), which is not yet commercially available because of the extended reconstruction time; these studies have showed relatively low specificity10. In contrast, IMR can be used in daily practice; however, while the good image quality of low-dose CCTA with IMR has been reported8, its diagnostic accuracy has not been well-established.Recently, low-radiation CCTA has been used to reduce radiation exposure with the low tube voltage method. However, increased noise is a problem of this technique, which can affect image quality and diagnostic performance. The newly introduced, knowledge-based iterative model reconstruction (IMR) with a fully iterative algorithm can now be clinically used to significantly reduce noise, while maintaining image quality, as compared to iterative reconstruction (IR) with tube voltage loweringHere, we aimed to assess the accuracy of CCTA using low tube voltage, the IMR technique, prospective ECG triggering, and a radiation dose of approximately 1 mSv, in comparison with ICA, in patients with suspected CAD. In particular, we evaluated the diagnostic accuracy on a per-segment, per-vessel, and per-patient basis, and determined its impact and the factors underlying misdiagnosis on CT images.In this study, we examined a total of 61 subjects . All CCTA procedures were performed without complications. The mean radiation dose of the 61 patients was 1.05 ± 0.36 mSv  were considered excellent (Grade 4), 293 segments (38.2%) were considered good (Grade 3), and 84 segments (10.9%) were considered adequate (Grade 2). None of the segments were considered non-diagnostic (Grade 1). The mean image quality was 3.40 ± 0.68. On per-segment analysis, the image quality of the RCA, LM, LAD, and LCx were found to be 3.70 ± 0.53, 3.90 ± 0.35, 3.41 ± 0.62, and 3.35 ± 0.71, respectively , whereas a reduction in the effective dose by 46% was observed in studies with scans performed at 100 kVp, as compared to scans performed at 120 kVp 14. Furthermore, in a previous study, we found that the use of 80 kVp yielded a reduction in the mean radiation dose by nearly 50% to sub-mSv levels, as compared to CCTA at 100 kVp15.However, the issue concerning the radiation dose during CCTA is important, considering the potential stochastic cancer risks associated with medical radiation exposure17. The relevant use of the lower kVp for CCTA as well as IMR can further provide reduced radiation doses without compromising image quality.IMR relies on statistical and system models, and views reconstruction as an optimization process. Newer techniques, such as the current one, may simultaneously provide further reduction in image noise, potential dose savings, and improved image quality under conditions of reduced tube voltage10.Our study with prospectively ECG-triggered CCTA at low kVp with the IMR technique showed clinically acceptable diagnostic accuracy, consistent with that in previous reports using other various reconstructive technologies for reducing radiation dose exposure. Based on our previous study, low-radiation CCTA using IMR is clinically feasible with appreciable image interpretability, and has the advantage of the executive time of reconstruction, as compared to other reports of the feasibility of submillisievert CCTA scanning with the latest-generation iterative reconstruction algorithms in clinical settings11. Previously reported CCTA with 80–100 kVp/150–210 mA scan protocol using MBIR showed 85% accuracy per vessel analysis9. Despite the reduced radiation dose (approximately 0.29 mSv), its extended reconstruction time causes difficulty in applying MBIR-CCTA in practice9.In the present study, low-radiation CCTA (1.05 ± 0.36 mSv) with low tube voltage (80 or 100 kVp) and IMR yielded high sensitivity (86.4%), specificity (96.1%), PPV (80.3%), and NPV (97.5%) in a per segment-based analysis. These results are comparable to previous findings for CCTA obtained from prospectively triggered studies using the IR algorithm with showing high diagnostic accuracy  in patients with suspected CAD20. Although low tube voltage settings can increase the blooming artifacts from calcification, under low radiation dose conditions, the use of IMR algorithms for suppressing blooming artifacts in the presence of coronary calcium can reduce the calcium scores and image noise, in comparison with the traditional filtered back projection (FBP)22. However, this factor may still represent the major cause of false positive CCTA results and image artifacts. Another factor affecting false positive results, which are misinterpreted as non-calcified plaques, is the streak artifact as a result of X-ray beam penetrability, which reduces the effective attenuation adjacent to the dense calcification. In the present study, false negativities were primarily affected by inadequate vessel attenuation at the distal small branches, as a result of the limited spatial resolution on MDCT that causes the blurring of vessel margins . The other 4 cases of false negative diagnoses were caused by eccentric calcifications on opposite sides of the vessel wall, which led to the underestimation of coronary artery stenosis severity, as reported in a previous study23.Our findings suggest that the diagnostic accuracy of CCTA for the detection of coronary artery stenosis was affected by various factors, similar to the previously reported factors for misdiagnosis on CCTA. The false positive results were primarily affected by the blooming artifact due to a greater extent of dense calcification in coronary arteries . The presence of densely calcified plaques accentuated the partial volume averaging of different densities within a single voxel, thus leading to worse coronary lumen visualization due to insufficient spatial resolution and consequently to overestimation of luminal stenosis24. This finding may be related to the fact that one diseased segment is sufficient for detection of CAD in a patient, and that this patient may have additional diseased segments undetected on CCTA. Thus, CCTA may be currently suitable for excluding CAD at the patient level, or for detection of stenoses at the segment level25.Based on the analysis of CCTA results from patients who underwent ICA, low-dose CCTA may be suitable for excluding CAD in low-to-intermediate probability populations from the favorable NPV on per-patient, per-vessel, and per-segment bases. Moreover, there was an inverse correlation between sensitivity and specificity in comparison of segment-based and patient-based analysis (86.4%/96.1% vs. 100%/83.3%), consistent with the findings of other meta-analyses27. Several reports have suggested that the contrast media dose at 80 kVp CCTA could be decreased via iodine enhancement, with acceptable image quality30. Low contrast-dose protocols could protect renal function by using a diagnostically appropriate amount of contrast medium for low-radiation CCTA. Therefore, there is a need for further investigation regarding low-radiation and low contrast-dose CCTA protocols with IMR reconstruction.With these low-radiation CCTA protocols, it is also possible to reduce the amount of contrast agent administered, because the iodine attenuation increases with a decrease in the tube voltage as a result of contrast enhancement by reducing the energy of X-ray photons, and the mean k-absorption edge of iodine is closer to the reduced voltage2) in this single-center study. Second, there might be a referral bias because patients for whom CCTA excludes a stenosis are rarely sent for follow-up angiography, which means that lesions could have been missed. However, in this retrospective observational study, we recruited patients who underwent ICA in accordance with the relevant clinical indications. Third, the effective radiation dose could be further reduced if the tube current of the patients was more strictly modulated. We used the CCTA scan protocol with 80 kVp/100–200 mAs or 100 kVp/100–150 mAs, according to the BMI of the patients. The use of the IMR techniques could reduce image noise and permit further reduction of the radiation dose, as data can be acquired using a lower tube current (<100 mAs)9. Although the use of lower tube current results in noisy images, IMR could reduce the image noise without reducing their diagnostic quality.This study has certain limitations. First, the main limitation is the small sample size, retrospective design and small body size of the study subjects  with a conversion coefficient of 0.014 mSv/(mGy × cm). There is a need for further investigation regarding use of the new cardiac-specific factors to ensure more accurate dosimetry; this can contribute to benefit-risk calculus in cardiac imaging strategies, and support optimization of radiation safety for patients.Recently, a new conversion factor for estimating effective doses during CCTA was presented, such that a cardiac-specific conversion factor of 0.026 mSv/(mGy × cm) is appropriate to estimate effective radiation doses; this reflects a new tissue-weighting factorIn conclusion, a low-radiation CCTA protocol with low tube voltage, low tube current, prospective ECG triggering, and IMR could be a useful strategy for reducing the radiation dose of CCTA, while maintaining the diagnostic accuracy. Our findings suggest that IMR enables the accurate non-invasive diagnosis of CAD with CCTA at a 1 mSv fraction of effective radiation.This study was approved by Gangnam Severance Hospital ethics committee/institutional review board (3-2016-0282) and the requirement for written informed consent was waived due to the retrospective nature of this study. All methods were performed in accordance with the relevant guidelines and regulations.The study was designed as a retrospective study. From January 2014 to February 2016, a total of 4078 patients with suspected CAD were referred to our hospital for ICA. Among these, 411 patients who underwent CCTA within 2 months prior to ICA were examined; 314 patients were excluded because they underwent CCTA at 120 kVp or because they underwent CT with retrospective ECG-gated data acquisition. Moreover, we excluded patients who underwent CCTA using reconstructive methods such as traditional FBP or hybrid IR (n = 4), or those who underwent CCTA for stent evaluation (n = 16), bypass graft evaluation (n = 15), or both (n = 1). Finally, a total of 61 patients  were enrolled in our analysis  with prospective ECG triggering and a step-and-shoot axial scanning technique. CT scanning was performed from the carina to the diaphragm in a craniocaudal direction at the end-inspiratory pause. When the pre-examination heart rate was higher than 65 beats per min, a β-blocker  was administered per os 1 hour before the examination, without additional intravenous injection.2 and 80 kVp for BMI < 25 kg/m2); detector collimation with dynamic z-focal spot imaging, 64 × 0.625 mm; gantry rotation time, 400 ms; table feed per rotation, 4 cm; and center of the imaging window, 70–80% of the R-R interval.The CT scanning parameters were as follows: tube voltage and tube current, 80 kVp and 100–200 mAs or 100 kVp and 100–150 mAs , was administered intravenously via an antecubital 18-gauge catheter at a rate of 0.50–0.75 mL/kg for 15 s, followed by the administration of 50 mL of normal saline using a power injector . Data acquisition was started 7 s after the trigger threshold of 130 HU was achieved at the proximal descending aorta using the real-time bolus tracking technique. All patients performed the breath-hold maneuver successfully. The effective radiation dose for CCTA was calculated by multiplying the dose-length product (DLP) with a conversion coefficient of 0.014 mSv/(mGy × cm).CT images were reconstructed using iterative model reconstruction . The images were sent to a picture archiving and communication system . The reconstruction parameters were as follows: slice thickness, 0.9 mm; increment, 0.45 mm; field of view, 15–23 cm; pixel image matrix, 512 × 512; and kernel, XCC. Post-processing and reconstruction were performed with multiplanar reformatted (MPR), curved planar reformatted (CPR), and maximum intensity projection (MIP) images using commercially available software .32. The segments were included for analysis only if the diameter of the origin was equal or larger than 1.5 mm.All images were reviewed and interpreted using the PACS workstation. Two radiologists (with more than 10 years of experience interpreting cardiac images) who were blinded to patient history and the results of coronary angiography evaluated the CCTA findings. Coronary arteries were divided into 18 segments according to the Society of Cardiovascular Computed Tomography guidelines2 at the ascending aorta. Image noise was evaluated in CCTA based on one standard deviation of the attenuation value at the ascending aorta. The SNR was calculated as vascular attenuation/image noise. The CNR was calculated as [(attenuation of vessel) - (attenuation of the adjacent perivascular fat)]/image noise.The vascular attenuation values were measured using a round region of interest larger than 1.5 cm23.The qualitative image quality of the 18 coronary segments was analyzed using a 4-point grading scale as follows: Grade 1 (poor/nondiagnostic), severe image degradation or discontinuation of vessel contour that prevented vessel lumen evaluation; Grade 2 (adequate), moderate image degradation that impeded vessel lumen evaluation; Grade 3 (good), minor image degradation that did not affect vessel lumen evaluation; and Grade 4 (excellent), no image degradation All reconstructed images, including axial source images and MPR, CPR, and MIP images, were analyzed to evaluate stenosis. Half of the sum of the normal proximal and distal lumen within the same segment was used as the reference for percent stenosis. Luminal narrowing greater than 50% was regarded as significant stenosis. If there was more than one lesion within the same segment, the worst stenosis percentage was used for the analysis. The results of CCTA were compared to those obtained with ICA. Subsequently, two radiologists re-evaluated the cases for false positive and false negative segments, and the possible factors affecting diagnostic accuracy, such as location, vessel wall condition, coronary calcium scores, vessel attenuation, vessel wall motion, and other possible factors, in each corresponding segment, were discussed. Discrepancies, if any, were resolved by consensus.ICA was performed with standard techniques via radial approach. All images were digitalized for offline analysis using commercially available software . Segments were included for analysis at the bifurcation area only if the diameter was equal to or larger than 1.5 mm. Using at least 2 image planes, quantitative measurements were performed as follows: degree of stenosis (%) = [/the reference diameter]*100. The reference diameter was determined as the mean of the normal proximal and distal lumen within the same segment. Each vessel was considered to have significant stenosis if the stenosis was greater than 50%.Continuous variables are expressed as mean ± standard deviation and categorical variables are expressed as frequencies or percentages. Shapiro-Wilk tests were performed to evaluate the distribution of the data Inter-reader agreement for the image quality assessment of CCTA was evaluated by using κ statistic. A κ value of 0.00–0.20 signified none to slight agreement; 0.21–0.40, fair agreement; 0.41–0.60, moderate agreement; 0.61–0.80, good agreement; and 0.81–1.00, excellent agreement. The sensitivity, specificity, PPV, NPV, and diagnostic accuracy of low-dose CCTA were calculated by using ICA as a reference, on a per-segment, per-vessel, and per-patient basis. All statistical analyses were performed with the SPSS 20 Statistical Package for the Social Sciences .Supplementary Tables and Figure


Enterococcus sp. has the capacity to produce dopamine in a gastrointestinal-like environment when supplied with the dopamine precursor L-3,4 dihydroxyphenylalanine (L-dopa). The results presented herein provide a means to select probiotics based on neurochemical-producing potential and suggest the possibility that probiotics containing E. faecium may serve to influence the host through dopaminergic pathways.The mechanisms by which probiotics may influence host physiology are still incompletely understood. Microbial endocrinology, a field representing the union of microbiology, endocrinology and neurobiology, has theorized that microorganisms have the capacity to serve as neurochemical delivery vehicles . Accordi Enterococci, including E. faecium and E. faecalis are natural members of the gastrointestinal flora and found in fermentation products. E. faecium is capable of producing a variety of complex molecules from aromatic compounds which influence the flavor of foods to the production of bacteriocins which show antibacterial activity against human pathogens such as Listeria initial) x 100.Conversion efficiency (C.E.) was calculated by the equation: . Inoculation was achieved by mixing 4.9mL of media with 100μL of bacterial suspension. Inoculated samples were grown for 24 hours at 37°C, anaerobically while being subjected to low speed (100rpm) magnetic stir bar mixing. Samples were then processed by a standardized approach for catecholamines [A diet containing 7.5% Mucuna powder (by dry weight) was prepared by mixing 4.9 grams of Mucuna powder  with 60 grams of feed . This entirety of this mixture was digested per the reviously. The proolamines . All conin vitro digestion of foodstuff enables the evaluation of neurochemical production potential by constituents of the microbiome [in vitro growth of bacteria in an environment that is reflective of the small intestinal host-based milieu [Enterococci to convert L-dopa to dopamine.We recently demonstrated that a simulated intestinal medium produced by the crobiome . Throughcrobiome , one cand milieu . By usin7 CFU/mL). All other strains were able to achieve growth on the order of 1.0x108 CFU/mL with strains such as ML1089 achieving growth over a full order of magnitude greater than ML1087. Isolate-to-isolate variability in the production of neurochemicals may account for the variability in achievable clinical outcomes that are encountered with the use of different probiotic preparations [The strain ML1082 consistently demonstrated the highest level of dopamine production . Otherwise, there was a high level of variation in the capacity to reproduce in the gastrointestinal-like SSIM and produce dopamine . ML1088 E. faecium to produce physiologically relevant amounts of the neurochemical dopamine from dietary substrates. While it has been known for decades that probiotics, such as Lactobacillus plantarum, produce neurochemicals including acetylcholine [E. faecium strains with demonstrated in vitro capacity to produce large amounts of dopamine to be administered to animals coupled with a food-based source of L-dopa such as Mucuna powder, to examine whether dopamine production occurs in vivo. If such experiments do, in fact, show that the administration of a synbiotic preparation of E. faecium plus an L-dopa source (such as a food-based one) leads to changes ascribable to dopaminergic pathways, then it is conceivable that dopaminergic modulation is a mechanism through which neurochemical-producing probiotics, such as E. faecium, may influence the host and modulate pathological processes such as inflammation which involve immune cells known to be responsive to neurochemicals [The present study demonstrates the hitherto unknown ability of lcholine , relativhemicals .in vivo will require carefully designed studies assessing physiological parameters of host cells grown in the presence of E. faecium. However, a review of the literature suggests that at least some of the purported effects of dopamine in the gastrointestinal tract are consistent with the effects observed when taking to E. faecium-containing probiotics. To substantiate, it is increasingly recognized that dopamine possesses an important immunoregulatory role and has the capacity to serve as a mediator enabling cross-system communication between the nervous and immune systems. Dopamine receptors are found throughout the immune system, one such example includes the demonstration that D1, D2, D3, D4 and D5 receptors have all been found in leukocytes [2 and D3 receptors in T cells activates T cell receptor-induced cell proliferation, and secretion of IL-2, IFN-γ and IL-4 [E. faecium. For example, it has been shown in broiler chickens that serum levels of the proinflammatory cytokines IL-1β, IL-2, IL-6 andIFN-γ, as well as the anti-inflammatory cytokines IL-4, IL-10, changed in response to the per oral administration of E. faecium strain NCIMB 11181 [As has been discussed in a previous report which first proposed the use of neurochemical-producing probiotics in the treatment of health and disease as well as behavior through the microbiota-gut-brain axis , confirmukocytes  and it hand IL-4 . This ovMB 11181 .E. faecium to modulate the immune system and promote the secretion of anti-inflammatory IL-4, it follows that both of these agents might be worth investigating for their role in the management of inflammatory diseases. In fact, E. faecium probiotics have been shown to be beneficial in the management of IBD [E. faecium be responsible for this benefit? An association of reduced endogenous dopamine production and responsiveness with human IBD has been demonstrated [Given the apparent potential for both dopamine and t of IBD –12. Coulnstrated , 22. Furnstrated . More renstrated , overallnstrated .50 of 6.3μM. In contrast, in longitudinal muscle, dopamine can produce relaxation with an EC50 of 29.0μM [50 of maximal response in the distal colon was only 20.0μM which is below the 71μM production of E. faecium grown in unsupplemented media and vastly below the 1563μM E. faecium is capable of producing when L-DOPA is abundant (+, Cl− and water absorption as well as K+ and bicarbonate secretion throughout various regions of the GI tract [in vivo [E. faecium can reduce the occurrence of diarrhea in animals [E. faecium may be a mechanism by which probiotics influence the water content of gastrointestinal material and alleviate diarrhea.In addition to immunomodulation, dopamine has the capacity to influence intestinal motility, intestinal secretion and water absorption. In circular muscle, dopamine can induce contractions with an ECf 29.0μM . Notablyabundant . DisruptGI tract  and dopa animals –32. BecaIn vivo, gastrointestinal L-dopa availability is likely influenced by intestinal absorption and dietary factors [Mucuna pruriens (Vicia faba cv Alameda can reach L-dopa concentrations as high as 6.75% of the total dry mass [E. faecium closely mirrored the availability of L-dopa. Though L-dopa is common in many raw plants and plant derived foodstuffs, L-dopa can degrade into various quinones under alkaline and thermal conditions[ factors . We thuspruriens . In a didry mass . The genonditions. This suEnterococci, would be the concern that a particular strain contains virulence traits or antibiotic resistance elements like vancomycin resistance. While these concerns are warranted, Enterococci are ubiquitous in the environment and these concerns should not preclude the use of an appropriately tested strain or prevent the search for other bacteria which may possess the capacity for dopamine production. The capacity to produce dopamine from L-dopa was common in all of our viable tested strains of E. faecium and may be for other species as well. Though the capacity to produce dopamine appears to be common trait in E. faecium, the conversion efficiency of L-dopa to dopamine varied greatly among isolates (E. faecium are equally suitable as probiotics.When considering the efficacy of a given probiotic, one must consider the specific strain behaviors. Of high importance with isolates . This imin vitro. No matter how complex the simulated media, the dynamic environmental changes that occur in a complex microbiota constantly regulated by the host cannot be fully replicated with this approach. In a polymicrobial community we may expect that some resources would be depleted, whereas others could be liberated. We may also expect that some metabolic processes could be regulated by the presence of signaling factors inherent to the in vivo environment. These limitations necessitate in vivo follow up in order to determine the physiological significance of dopamine production by Enterococci. Limitations aside, sSIM media does allow one to provide microorganisms with an environment containing resources consistent with those made available from a host’s dietary intake and non-microbial processing. The E. faecium isolates tested here come from a variety of animals and anatomical sites; they demonstrate a wide range in the capacity to produce dopamine. This work makes it apparent that Enterococci possess the enzymatic capacity to produce dopamine and that they can do so with resources derived from the diet. Of note, E. faecium did not appear to produce dopamine from tyrosine but only from dietary sources of L-dopa. In sSIM tyrosine is regularly available in millimolar quantities [Enterococci are well known for their capacity to convert tyrosine to tyramine because of the enzyme tyrosine decarboxylase [E. faecium is converted to tyramine [In vivo it is conceivable that in addition to dietary sources of L-dopa, L-dopa produced by other microbes may facilitate dopamine production.We acknowledge that a drawback of the present work is that it is relies predominantly on the behavior of a single species isolates grown antities . Howeverboxylase  and our tyramine . In vivoE. faecium, one must also make dietary provision of L-dopa, effectively creating a synbiotic. Together these findings suggest that probiotics, like classic pharmacological interventions, may require standardized formulations and delivery routines in order to provide optimal benefits. The results presented herein demonstrate that utilization of a microbial endocrinology-based method may therefore provide for mechanistic driven approaches to identify probiotics that may be useful to treat human disease where neurochemical pathways are known to play a role. Moving forward, studies assessing the ability of E. faecium ability to modulate host tissues should also consider the availability of L-dopa and consider measuring physiological and immunological parameters known to be associated with dopaminergic regulation.Dietary variations in L-dopa availability and the presence of other potential commensal microorganisms which can deliver L-dopa may provide a rationale for the varied response individuals taking probiotics achieve. It is conceivable that in order to achieve a benefit with


We investigated protein profiles specific to vascular lesions mimicking Kaposi sarcoma (KS), based on stepwise morphogenesis progression of KS. We surveyed 26 tumor-associated proteins in 130 cases, comprising 39 benign vascular lesions (BG), 14 hemangioendotheliomas (HE), 37 KS, and 40 angiosarcomas (AS), by immunohistochemistry. The dominant proteins in KS were HHV8, lymphatic markers, Rb, phosphorylated Rb, VEGF, and galectin-3. Aberrant expression of p53, inactivation of cell cycle inhibitors, loss of beta-catenin, and increased VEGFR1 were more frequent in AS. HE had the lowest Ki-67 index, and the inactivation rates of cell cycle inhibitors in HE were between those of AS and BG/KS. Protein expression patterns in BG and KS were similar. Clustering analysis showed that the 130 cases were divided into three clusters: AS-rich, BG-rich, and KS-rich clusters. The AS-rich cluster was characterized by high caveolin-1 positivity, abnormal p53, high Ki-67 index, and inactivated p27. The KS-rich cluster shared the features of KS, and the BG-rich group had high positive expression rates of galectin-3 and low bcl2 expression. In conclusion, although the rate was different, AS and HE tended to have less cell cycle marker expression than KS, and features of BG and activated KS cell signaling were similar. Kaposi sarcoma (KS) is an intermediate grade, rarely metastasizing vascular tumor caused by human herpes virus 8 (HHV8) infection . AccordiVascular tumors in skin and soft tissues are generally classified into four categories: benign; intermediate, locally aggressive; intermediate, rarely metastasizing; and malignant lesions, according to their clinical behavior and morphological features . AlthougIn the present study, we aimed to determine specific protein profiles in benign, intermediate, and malignant vascular tumors by assessing the expression of 26 proteins. Using this method, we aimed to enable the differential diagnosis of vascular lesions, and compare the differences and similarities in pathophysiology between each tumor group and KS. p < 0.001). D2-40 and podoplanin expression rates were also higher in KS compared to those in other groups . Positive rates of CD31 and CD34 were relatively high in all groups, but CD34 expression was lower in AS compared with that in other groups . With the exception of KS, each group expressed D2-40 and podoplanin in some areas, implying lymphatic differentiation. However, there were no significant differences in expression of these proteins between BG, HE, and AS. p = 0.008) and AS group . Interestingly, the BG and AS groups showed similar high Ki-67 index levels (p < 0.091). The Ki-67 index of the KS group (19.5 ± 18.3) was in between that of the HE and BG/AS groups, although there was no statistical significance  and granuloma pyogenicum (12/18); Ki-67 expression levels in these lesions were 46.33% ± 16.84% (mean ± SD). BG lesions with Ki-67 expression below the median value included acroangiodermatitis, angiofibroma, cavernous hemangioma, cherry angioma, hemangioma, intravascular histiocytosis, and stasis dermatitis; Ki-67 expression levels were 8.71% ± 8.10% (mean ± SD).p < 0.001 and p = 0.009, respectively, p = 0.012 and p < 0.001, respectively). The rates of inactivated p27 in the HE group were also higher than those in the BG and KS groups . Aberrant p53 expression  was found only in the AS and HE groups, with no positive KS and BG cases. Positive expression of cell survival markers was highest in AS and lowest in HE. The BG and KS groups showed medium levels of cell survival marker expression that were between those of the AS and HE groups. Bcl2 and NF-κB were more upregulated in KS than in AS. Positive expression rates and p-values are listed in detail in Among 10 proteins involved in cell cycle, apoptosis, and cell survival, Rb and phosphorylated Rb (pRb) were more highly expressed in the KS group , when compared to their expression in BG, HE, and AS . C-kit was exclusively expressed in AS (4/40 [10%]). However, expression of VEGF was slightly higher in the KS and AS groups than in BG and HE, but this difference was not significant .Out of the four tumor groups, AS had the highest positive expression of VEGFR1 . Conversely, caveolin-1 expression was higher in HE and AS groups  than in BG and KS . Other markers that showed differential expression in specific tumor groups included CD44, EMA, and β-catenin. CD44 was uniquely downregulated in KS . Positive expression of EMA was uniquely found in AS (4/40 [10%]), and total loss of β-catenin expression was more pronounced AS (15%) than in the other groups .Comparative analysis of cell adhesion and motility-associated proteins showed that positive rates of galectin-3 in BG and KS groups  were higher than those in HE and AS groups . Hierarchical clustering showed three large clusters a. The diThe 2013 World Health Organization (WHO) classification of vascular tumors in soft tissue and bone comprises four groups of vascular tumors: benign, locally aggressive intermediate, rarely metastasizing intermediate, and malignant tumors . HemangiHE was characterized by having the lowest Ki-67 index in the four groups, which was a difference large enough to enable differentiation between HE and the other tumor types. This feature can also be helpful to differentiate AS or BG tumors. However, the use of the Ki-67 index for differentiation between benign and vascular tumors is controversial ,14. In oCaveolin-1 and galectin-3 showed contrasting expression levels in AS/HE and BG/KS. Galectin-3 was more frequently expressed in BG and KS groups, whereas caveolin-1 showed the opposite. Caveolin-1 is a scaffolding protein found in the plasma membrane, which mediates transport of intracellular and extracellular proteins and signal transduction. Differential expression levels of caveolin-1 have been correlated with myogenic differentiation of rhabdomyosarcoma . GalectiCD44 is a blood vascular endothelium-specific hyaluronan receptor . It is uCluster analysis was used to investigate specific characteristics in each group. Out of the 130 cases, 62% (81) were successfully classed into three clusters. However, the remaining 38% cases were distributed within other clusters. The most accurate clustering was that of the KS group (98%), whereby only two cases were misplaced. We tried to determine the clinicopathologic difference between clustered cases and non-clustered cases , but the small size of groups was a limitation for this analysis. Interestingly, the cluster pattern and protein expression differed according to tumor location . Five caIn conclusion, we were able to find patterns that differentiated BG, HE, KS, and AS by comparing the expression of 26 tumor-associated proteins. BG and KS shared a “hyperplastic pattern” marked by activated proliferative features, even though the underlying pathogen was different. HE and AS shared aberrant expression of cell cycle inhibitors, despite the differences in their tumor grade. Thus, vascular tumors mimicking Kaposi sarcoma can be categorized into three clusters: cluster 1, altered expression of tumor suppressor genes  with a high proliferative index, enriched AS; cluster 2: high CD44 expression with a low proliferative index suggesting stem cell differentiation, mixed diagnosis; and cluster 3: lymphatic differentiation with activated Rb pathway, enriched KS.We selected vascular neoplasm patients within surgical pathology archives, covering the period between 1999 and 2009, from the Seoul National University Hospital and Seoul National University Bundang Hospital. The BG group comprised only patients with skin lesions, whereas the HE, KS, and AS patients were selected with no site limitation due to low incidence rates, compared to those of BG. We retrospectively enrolled 130 cases from a total of 107 patients, including 39 BG cases/patients, 14 HE cases from 11 patients, 37 KS cases from 26 patients, and 40 AS cases from 28 patients. For all cases included in the study, medical records and paraffin block tissue samples were obtained. The detailed pathological diagnosis of each study group is listed in Four-µm-thick tissue sections on glass slides were deparaffinized at 60 °C for 1 hour, dewaxed at 72 °C for 3 minutes, and hydrated by washing with alcohol three times. Immunohistochemical staining was performed using two types of autostainer, Leica Bond-max autostainer  and Ventana BenchMark XT staining system . Antigens were retrieved by heat treatment in the epitope retrieval solution for each autostainer . After peroxidase blocking for 5 minutes, the primary antibody was incubated for 15 minutes. The type, dilution level, and company information of the 25 primary antibodies used are listed in Immunohistochemical analysis was performed according to previously described criteria and methods generally accepted by pathologists . Firstlyp-values were less than 0.05. A chi-square test or Fisher’s exact test (two-sided) was performed to determine differences in expression between markers and disease groups. A nonparametric Mann–Whitney U test or median test was performed to compare Ki-67 labeling index means and the median value of age. All statistical analyses were conducted using IBM SPSS Statistics 21.0 . Hierarchical clustering of the 130 samples was performed based on the expression rates of the 25 markers, excluding HHV8. A standard Pearson correlation was used as the distance of two samples, and complete linkage clustering was used. Clustering analysis and drawing were performed using the Gene Cluster 3.0 open source clustering software and Java TreeView 1.1  . The res


It was found that the PVP concentration was one of the most crucial processing parameters determining the final properties of WO3 nanofibers. The optimum concentration of PVP was from 75 to 94 g L−1. The average diameter of the nanofibers increases with increasing the PVP concentration, whereas it is decreased after sintering and orthorhombic structure were formed at 500 °C. The photocatalytic properties of the as-synthesized nanofibers were also investigated by degrading methylene blue and twofold efficiency was obtained compared with that of commercial WO3 microparticles.The present study involves the fabrication of tungsten trioxide (WO The textile industry effluent is a major source of water pollution which can destroy the aquatic life and influence world climates change seriously . The maiCurrently, different strategies have been employed for treating the textile effluents before they are released in water bodies including microfiltration , chemica3) is one of the most popular metal oxides which have been extensively used as electrochromic devices and gas sensor for water splitting [3 has a broad range of band-gap values which ensures considerable photocatalytic and photoelectrocatalytic activity under visible-light illumination [3-based materials as well as their photocatalysis have attracted much attention, for example, nanoparticles for degradation of lidocaine under visible and sunlight irradiation [3, for example, photo-deposition of platinum in WO3 nanostructures [3 nanotubular composite fabricated by electrochemical method [3 [3 nanofibers with large aspect ratio are rarely reported before [3 such as nanorods [Tungsten trioxide /citric acid/tungstic acid. The photocatalytic efficiency of the as-synthesized nanofibers was also investigated via degrading methylene blue.Generally, nanofibers are produced by three techniques of self-assembly , phase s-sensing , core–shatalysis , polycryatalysis , templatatalysis , and impatalysis . Especianofibers , 37, 39.Mw = 1,300,000 was purchased from Sigma Aldrich in USA. Commercial WO3 powders (mean particle size 30 μm) were obtained from Kanto Chemical Company and methylene blue (99 % purity) from Showa Chemical Company in Japan. Ammonia solution was from Duskan Pure Chemical Company Ltd, and citric acid monohydrate (99.5 % purity) was from Samchun Pure Chemicals in Republic of Korea.PVP of 3 nanofibers were synthesized using an electrospinning method. Commercially available WO3 powders were dissolved in hot ammonia solution with stirring firstly. After the mixture was cooled down to room temperature, the insoluble materials were removed by filtration after which an amount of citric acid was added as chelating agent for binding WO3 [−1) were added into the above solution followed by magnetic stirring for about 1 day. The weight ratio of PVP/citric acid/WO3 was kept constant at a ratio of (1.1:2.7:1.0). The mixed solution was aged on hot plate for different time to form different concentration of PVP/citric acid/tungstic acid precursor gel for electrospinning. In this work, three kinds of precursor solution were prepared in which the concentration of PVP was 75, 94, and 140 g L−1, respectively.The WOding WO3 . Then, P−1 by using a syringe pump . The syringe was aligned perpendicular to flat bed collector of electrospinning apparatus. The wire originated from the positive electrode (anode) was connected with the needle tip of plastic syringe with an alligator clip. Finally, the solutions were electrospun at +18 kV. The distance between the needle tip and the flat bed metallic collector covered with aluminum foil was kept at 15 cm. The as-spun nanofibers were vacuously dried at 60 °C for 24 h in the presence of P2O5 to remove the residual moisture from used solvents. Further on, the samples were subjected to calcination at 500 °C for 2 h with a heating rate of 1 °C min−1.A high voltage power supply  capable of generating voltages up to 30 kV was used as the electric field source for spinning of nanofibers. Solutions to be electrospun were supplied through a 20-mL plastic syringe attached with stainless steel having 22 gages, (0.7 mm OD × 0.4 mm ID). The feeding rate of solution to be electrospun was adjusted to be 0.2 mL min−1 with a heating rate of 20 °C min−1. Information about the phases and crystallinity of nanofibers was obtained using X-ray diffractometer  with Cu, Cr (λ = 1.540 A) radiation over Bragg angle ranging from 20° to 60°. For photocatalytic property, the samples were suspended (1 g L−1) in methylene blue solution (20 ppm) in a batch reactor and illumination was applied using a commercial haloline visible-light illumination . The evolution of the methylene blue concentration was measured by UV–Vis spectrometry  following its 660 nm characteristic band.The morphology of the samples was examined using a scanning electron microscope . The samples were coated with a thin layer of platinum–palladium for 50 s with the discharge current of 10–15 mA for three reparative cycles using  ion sputter. After coating, the micrographs were taken at an accelerating voltage of 25 kV, and images were captured with magnifications of 1 and 20 K. The thermal properties were tested by thermogravimetry/differential scanning calorimetry instrument (SETARAM TG/DSC 92). The samples were heated from 30 to 600 °C under a continuous nitrogen purge of 20 mL min−1), electrospinning process could not be carried out due to low viscosity of precursor solutions (data not shown). However, when PVP concentration was 75 g L−1 or higher, the electrospinning was highly achievable for nanofibers. Figure −1 of PVP. From Fig. −1 .In order to confirm the precursor was completely removed after calcination, TGA of pure PVP, citric acid, and WO3 nanofibers by degrading methylene blue under visible-light illumination. For comparison, the commercial powders (mean particle size of 30 μm) were also investigated simultaneously. After irradiation for 2 h, the degradation efficiency for commercially available WO3 powders was only about 23 %, while the degradation efficiency for WO3 nanofibers (PVP: 140 g L−1) was 27 %, (PVP: 94 g L−1) 46 %, and (PVP: 75 g L−1) 50 %, respectively. The degradation efficiency for nanofibers is much higher than that of commercially available WO3 particles of which mean diameter is 30 μm according to manufacturer. It can be observed that the degradation efficiency for nanofibers with the smaller diameter prepared from (PVP: 75 g L−1) was 50 % which is higher than other nanofibers combinations with lager diameter. Therefore, it can be concluded that electrospinning WO3 nanofibers with smaller diameters can significantly improve the photocatalytic activity than commercially available WO3 micro-powders.Figure 3 nanofibers were fabricated by an electrospinning technique using PVP/citric acid/tungstic acid as precursor solutions. The average diameter of nanofibers is 80–100 nm and increases with PVP concentration. After sintering at 500 °C for 2 h, the diameter was reduced and the structure was transferred from amorphous to orthorhombic. The photocatalytic activity of WO3 nanofibers was found much higher than that of commercial WO3 powders in degradation of methylene blue under visible-light illumination.WO