Residual Viability Research Articles

Antimicrobial photodynamic therapy effectively reduces Porphyromonas gingivalis infection in gingival fibroblasts and keratinocytes: An in vitro study

BackgroundPorphyromonas gingivalis possess the ability to invade host cells which prevents this pathogen from eradication by conventional periodontal therapy. Recently, antimicrobial photodynamic therapy (aPDT) was introduced to periodontal treatment as a complementary antibacterial method. The aim of this study was to evaluate the effect of toluidine blue-O (TBO) mediated aPDT on the viability of P. gingivalis invading gingival fibroblasts and keratinocytes in an in vitro model of infection. MethodsPrimary human gingival fibroblasts (PHGF) and telomerase immortalized gingival keratinocytes (TIGK) were infected with Pg ATCC 33277. Two concentrations of TBO (0.01 mg/mL, TBO-c1 and 0.001 mg/mL, TBO-c2) and a non-laser red light source (λ = 630 nm) were applied to treat both cell-adherent/intracellular Pg (the adhesion/invasion model) or exclusively the intracellular bacteria (the intracellular infection model). ResultsThe median viability of cell-adherent/intracellular Pg in infected keratinocytes declined from 1.88 × 105 cfu/mL in infected cells treated with TBO without irradiation to 40 cfu/mL upon irradiation for 10 s with TBO-c1. At higher light doses a complete photokilling of P. gingivalis was observed. Pg from exclusively intracellular infection model was also efficiently eradicated as the residual viability dropped from 1.44 × 105 cfu/mL in control samples to 160, 20 and 10 cfu/mL upon irradiation for 10, 20 and 30 s, respectively. In the infected fibroblasts irradiation significantly reduced bacterial viability but did not completely eradicate the intracellular pathogen. ConclusionsAntimicrobial PDT is effective in reducing the viability of intracellular periopathogens, however those residing within gingival fibroblasts seems to attenuate the photokilling effectiveness of this method.

Contrast-enhanced ultrasound (CEUS) in HCC diagnosis and assessment of tumor response to locoregional therapies.

Hepatocellular carcinoma (HCC) is a global problem constituting the second leading cause of cancer deaths worldwide, thereby necessitating an accurate and cost-effective solution for managing care. Ultrasound is well poised to address this need due to its low cost, portability, safety, and excellent temporal resolution. The role of ultrasound for HCC screening has been well established and supported by multiple international guidelines. Similarly, contrast-enhanced ultrasound (CEUS) can be used for the characterization of focal liver lesions in high-risk populations, and standardized criteria for CEUS have been established by the American College of Radiology Liver Imaging Reporting & Data System (LI-RADS). Following HCC identification, CEUS can also be highly beneficial in treatment planning, delivery, and monitoring HCC response to locoregional therapies. Specific advantages of CEUS include providing real-time treatment guidance and improved diagnostic performance for the detection of residual tumor viability or recurrence, thereby identifying patients in need of retreatment substantially earlier than contrast-enhanced CT and MRI. This review provides a primer on ultrasound and CEUS for the screening and characterization of HCC, with an emphasis on assessing tumor response to locoregional therapies.

39800 Immune Checkpoint Blockade during Periprosthetic Joint Infection

ABSTRACT IMPACT: If immune checkpoint blockade increases bacterial clearance with or without antibiotics in vitro, clinical application would be almost immediate and dramatic creating a seismic shift in the current therapeutic paradigm of periprosthetic joint infection. OBJECTIVES/GOALS: Periprosthetic joint infection (PJI) is a major cause of failure after joint replacement. Currently, the treatment of PJI relies on removing biofilm contaminated implants. Some of the bacteria within biofilm undergo a phenotypic shift becoming small colony variants (SCVs). SCVs induce local immunosuppression through PD-1/L1 signaling. METHODS/STUDY POPULATION: We will infect cultured human macrophages and bone marrow aspirate with stable Staphylococcus aureus SVCs and treat with anti-PD-1 or anti-PD-L1 monoclonal antibodies with and without antibiotics (e.g., gentamycin, cefazolin, vancomycin, rifampicin) and assess the residual bacterial viability. We will utilize multiplexed ion beam imaging to quantify PD-1/L1 expression in human tissue from patients with a chronic PJI and compare those to patients undergoing an aseptic revision. Patients with a chronic PJI are likely to have increased expression of PD-1/L1 as their tissue samples are prospectively screened. RESULTS/ANTICIPATED RESULTS: SCVs reduce the phagocytic activity of macrophages and can survive intracellularly. SCVs also induce anti-inflammatory M2-macrophage polarization and recruit a heterogeneous group of immature monocytes and granulocytes called myeloid-derived suppressor cells (MDSC) to the periprosthetic microenvironment. M2-macrophages and MDSCs then produce an immunosuppressive cytokine milieu characterized by increased IL-10 and decreased TNF-α. Clinically isolated SCVs up-regulate the expression of PD-L1 and PD-L2 on the surface of macrophages, representing a mechanism by which SCVs induce host immunosuppression and survive immune clearance. Our preliminary data show PD-L1 expression during septic PJI, but not in aseptic revisions. DISCUSSION/SIGNIFICANCE OF FINDINGS: If immune checkpoint blockade is shown to increase bacterial clearance with or without antibiotics, host immunomodulation would represent a novel class of therapeutic adjuvants to assist surgical debridement and antibiotic administration that could be superimposed on existing treatment algorithms to improve PJI related outcomes.

Abstract PO025: Development of patient derived organoid cultures of two advanced stage serous carcinomas of the uterus

Abstract Advanced stage serous carcinoma of the uterus typically portends a poor prognosis. There is an urgent need to augment clinical trial data with different methods of treatment evaluation before a patient is made to try and potentially fail a therapy. Patient derived organoids (PDOs) may represent such an opportunity. We describe the successful culture and drug sensitivity testing of two PDOs from patients with advanced stage serous carcinoma of the uterus who received neoadjuvant chemotherapy. Clinical response to treatment was compared with the organoid response to the same regimens. Drug sensitivity data are reported as percentage of residual cell viability 48 hours after one standard treatment dose, in which 100% viability indicates no response and 0% viability reflects complete cell death. We hypothesized that significant cell killing, i.e. a low cell viability in our assay, would be required to achieve clinical drug sensitivity in patients. Both patients were clinically platinum resistant. Patient 1 had a large burden of disease at the time of presentation, including skeletal and liver metastasis. Though she initially partially responded to triplet therapy of paclitaxel, carboplatin and bevacizumab, she ultimately had persistent unresectable disease after 6 cycles of therapy. The percent residual, post-drug cell viability for PDO-1 was 45% in response to paclitaxel, carboplatin and bevacizumab combinatorial treatment, 35% in response to paclitaxel and carboplatin, and 100% in response to single agent bevacizumab. Patient 2 received paclitaxel, carboplatin and trastuzumab because her tumor was positive for HER2neu. Though she also had an initial response to treatment, she experienced a recurrent malignant pleural effusion 3 months after completing platinum-containing triplet (trastuzumab was continued as a maintenance therapy). The percent residual cell viability for PDO-2 was 51% in response to the combination of paclitaxel and carboplatin. Previous investigators have identified PDOs as a potentially powerful functional model for treatment response. We confirm that it is feasible to establish organoids and to rapidly test agents for cell sensitivity even prior to the first post-surgical chemotherapeutic treatment cycle. For both patients, the PDO models were only moderately sensitive to the agents used for therapy, and the lack of long-term benefit achieved for these patients may have been suggested a priori by the PDO models. However, further studies are necessary to establish a threshold percent value of post-treatment cell viability that is predictive of clinical patient response. We hypothesize that such a threshold appears to be significantly below 50% from these two initial cases. Citation Format: Andreea M. Newtson, Jianling Bi, Yuping Zhang, Eric Devor, Kristina Thiel, Kimberly K. Leslie. Development of patient derived organoid cultures of two advanced stage serous carcinomas of the uterus [abstract]. In: Proceedings of the AACR Virtual Special Conference: Endometrial Cancer: New Biology Driving Research and Treatment; 2020 Nov 9-10. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(3_Suppl):Abstract nr PO025.

Effective decellularisation of human saphenous veins for biocompatible arterial tissue engineering applications: Bench optimisation and feasibility in vivo testing

Human saphenous vein (hSV) and synthetic grafts are commonly used conduits in vascular grafting, despite high failure rates. Decellularising hSVs (D-hSVs) to produce vascular scaffolds might be an effective alternative. We assessed the effectiveness of a detergent-based method using 0% to 1% sodium dodecyl sulphate (SDS) to decellularise hSV. Decellularisation effectiveness was measured in vitro by nuclear counting, DNA content, residual cell viability, extracellular matrix integrity and mechanical strength. Cytotoxicity was assessed on human and porcine cells. The most effective SDS concentration was used to prepare D-hSV grafts that underwent preliminary in vivo testing using a porcine carotid artery replacement model. Effective decellularisation was achieved with 0.01% SDS, and D-hSVs were biocompatible after seeding. In vivo xeno-transplantation confirmed excellent mechanical strength and biocompatibility with recruitment of host cells without mechanical failure, and a 50% patency rate at 4-weeks. We have developed a simple biocompatible methodology to effectively decellularise hSVs. This could enhance vascular tissue engineering toward future clinical applications.

Noninvasive cardiovascular imaging for myocardial necrosis, viability, stunning and hibernation: evidence from an umbrella review encompassing 12 systematic reviews, 286 studies, and 201,680 patients.

Introduction The concomitant presence of myocardial necrosis with myocardial ischemia, stunning or hibernation may complicates appraisal of left ventricular (LV) function and patient management. Several imaging modalities have been proposed for the accurate assessment of myocardial necrosis, viability, stunning and hibernation, with mixed results. We aimed to review the evidence base on myocardial necrosis, stunning and hibernation by conducting an umbrella review (i.e. overview of systematic reviews). Evidence acquisition We searched PubMed and The Cochrane Library for meta-analyses focusing on the diagnostic, prognostic, or management appraisal of myocardial necrosis, viability, stunning and hibernation. Diagnostic test accuracy, prognostic yield, and clinical outcomes were systematically abstracted from shortlisted reviews. Evidence synthesis From an initial set of 6069 citations, 12 systematic reviews were finally included, encompassing 286 studies and 201,680 patients. Cardiac magnetic resonance imaging (CMR) had favorable results in 4 reviews that focused on the diagnosis of myocardial stunning or hibernation in patients followed for 6±4 months after coronary revascularization (sensitivity 96% and specificity 91%). Positron emission tomography (PET), single photon emission tomography (SPECT) and CMR in 6 meta-analyses had each a significant and independent prognostic role for the prediction of fatal and non-fatal cardiovascular events in patients with follow-up of 2.8±1.7 years. Finally, 2 reviews with 2.3±1.1 years of follow-up showed moderate quality evidence in favor of coronary revascularization in patients with objective signs of myocardial viability. Conclusions The appraisal of myocardial necrosis and residual viability remains a cornerstone of the modern management of patients with CAD. Current imaging modalities (echocardiography, PET, SPECT and CMR) are widely used. Further trials using contemporary methods are warranted to further clarify the impact of viability assessment on patient management, and the cumulative risk of morbidity and mortality.

Noninvasive cardiovascular imaging for myocardial necrosis, viability, stunning and hibernation: evidence from an umbrella review encompassing 12 systematic reviews, 286 studies, and 201,680 patients.

The concomitant presence of myocardial necrosis with myocardial ischemia, stunning or hibernation may complicates appraisal of left ventricular (LV) function and patient management. Several imaging modalities have been proposed for the accurate assessment of myocardial necrosis, viability, stunning and hibernation, with mixed results. We aimed to review the evidence base on myocardial necrosis, stunning and hibernation by conducting an umbrella review (i.e. overview of systematic reviews). We searched PubMed and The Cochrane Library for meta-analyses focusing on the diagnostic, prognostic, or management appraisal of myocardial necrosis, viability, stunning and hibernation. Diagnostic test accuracy, prognostic yield, and clinical outcomes were systematically abstracted from shortlisted reviews. From an initial set of 6069 citations, 12 systematic reviews were finally included, encompassing 286 studies and 201,680 patients. Cardiac magnetic resonance imaging (CMR) had favorable results in 4 reviews that focused on the diagnosis of myocardial stunning or hibernation in patients followed for 6±4 months after coronary revascularization (sensitivity 96% and specificity 91%). Positron emission tomography (PET), single photon emission tomography (SPECT) and CMR in 6 meta-analyses had each a significant and independent prognostic role for the prediction of fatal and non-fatal cardiovascular events in patients with follow-up of 2.8±1.7 years. Finally, 2 reviews with 2.3±1.1 years of follow-up showed moderate quality evidence in favor of coronary revascularization in patients with objective signs of myocardial viability. The appraisal of myocardial necrosis and residual viability remains a cornerstone of the modern management of patients with CAD. Current imaging modalities (echocardiography, PET, SPECT and CMR) are widely used. Further trials using contemporary methods are warranted to further clarify the impact of viability assessment on patient management, and the cumulative risk of morbidity and mortality.

Quantitative Hybrid Cardiac [18F]FDG-PET-MRI Images for Assessment of Cardiac Repair by Preconditioned Cardiosphere-Derived Cells

Cardiosphere-derived cells (CDCs) are progenitor cells derived from heart tissue and have shown promising results in preclinical models. APOSEC, the secretome of irradiated peripheral blood mononuclear cells, has decreased infarct size in acute and chronic experimental myocardial infarction (MI). We enhanced the effect of CDCs with APOSEC preconditioning (apoCDC) and investigated the reparative effect in a translational pig model of reperfused MI. Supernatants of CDCs, assessed by proteomic analysis, revealed reduced production of extracellular matrix proteins after in vitro APOSEC preconditioning. In a porcine model of catheter-based reperfused anterior acute MI (AMI), CDCs with (apoCDC, n = 8) or without APOSEC preconditioning (CDC, n = 6) were infused intracoronary, 15 min after the start of reperfusion. Untreated AMI animals (n = 7) and sham procedures (n = 5) functioned as controls. 2-deoxy-2-(18 F)-fluoro-D-glucose-positron emission tomography-magnetic resonance imaging ([18F]FDG-PET-MRI), with late enhancement after 1 month, showed reduced scar volume and lower transmurality of the infarcted area in CDC and apoCDC compared to AMI controls. Segmental quantitative PET images displayed indicated more residual viability in apoCDC. The left-ventricle (LV) ejection fraction was improved nonsignificantly to 45.8% ± 8.6% for apoCDC and 43.5% ± 7.1% for CDCs compared to 38.5% ± 4.4% for untreated AMI. Quantitative hybrid [18F]FDG-PET-MRI demonstrated improved metabolic and functional recovery after CDC administration, whereas apoCDCs induced preservation of viability of the infarcted area.

Contrast-Enhanced Ultrasound in the Short-Term Evaluation of Hepatocellular Carcinoma after Locoregional Treatment

Background: Contrast-enhanced ultrasound (CEUS) with second-generation contrast agents performed 1 month after hepatocellular carcinoma (HCC) treatment is almost as sensitive as contrast-enhanced computed tomography (CECT) in depicting the residual tumor. However, the efficacy of CEUS performed early after the procedure is still debated. Aim: We evaluated the diagnostic accuracy (DA) of CEUS for the assessment of tumor response shortly after locoregional therapy in patients with unresectable HCC. Methods: Ninety-four patients with 104 HCC lesions who were scheduled to receive percutaneous ethanol injection, radiofrequency ablation, transcatheter arterial chemoembolization, or combined treatment were enrolled in this study. With CECT at 1-month as the reference standard, the DA of CEUS performed 48-h after the procedure was evaluated. Patients were followed-up to look for tumor or disease progression. Results: Based on CECT findings, 43/104 lesions were diagnosed as having residual viability after 1 month. CEUS performed 48 h after treatment detected residual tumor in 34/43 nodules with treatment failure at CECT with a sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of 79.1, 96.7, 94.4, 86.8, and 89%, respectively. There was a high degree of concordance between CEUS and CECT (kappa coefficient = 0.78). A hyperemic halo was detectable in 35 lesions without a statistically significant difference between concordant and discordant cases. In patients with uninodular disease responders according to 48 h CEUS had a significantly longer mean overall survival and time to progression compared to nonresponders. Conclusion: CEUS performed 48 h after treatment can be considered a reliable modality for the evaluation of the real extent of necrosis and has prognostic value in the assessment of HCC.

Deferoxamine Protects Stromal/Stem Cells of "Lull pgm System"-Processed Lipoaspirates Against Damages Induced by Mitochondrial Respiration Inhibition.

The ischemic environment of the receiving area compromises the outcome of autologous fat grafts. The aim of this study was to isolate and expand the stromal vascular fraction from patient lipoaspirates and investigate the gain in cell viability exerted by some protective agents against the blockage of mitochondrial respiration. The aspirates were (1) washed, using the "Lull pgm system," (2) centrifuged and (3) decanted. The corresponding stromal vascular fractions were isolated, and after cell adherence selection, the stromal/stem cell subpopulations were exposed to Antimycin A for 1h. Then, the protection induced by cell pretreatment with deferoxamine, diazoxide and IGF-1 was evaluated. The residual cell viability of the "Lull pgm system"-washed samples was greater than that of the centrifuged samples (p < 0.05), and this advantage was maintained during the following 12days of culture. The administration of 400μM deferoxamine before Antimycin A treatment increased the number of viable cells from 56.5 to 80.8% (p < 0.05). On the contrary, the pretreatment with 250μM diazoxide or 0.1μg/ml IGF-1 did not exert any significant pro-survival action. Echinomycin abolished the positive effect of deferoxamine, suggesting that its protection involved HIF-1α. Adipose-derived stromal-stem cells survive the inhibition of mitochondrial respiration better if the lipoaspirate is washed using the "Lull pgm system" rather than centrifuged. Moreover, a significant contribution to cell survival can be obtained by preconditioning stromal-stem cells with deferoxamine. In a clinical perspective, this drug could be safely administered before surgery to patients undergoing autologous fat transfer. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Propidium monoazide–polymerase chain reaction for detection of residual periprosthetic joint infection in two-stage revision

False negative culture results in periprosthetic joint infection (PJI) are not uncommon particularly when patients have received long term antibiotics. Polymerase chain reaction (PCR) has a lower specificity partly due to detection of residual DNA from dead bacteria. Propidium monoazide (PMA) prevents DNA from dead bacteria from being amplified during the PCR. This study aimed to determine the role of PMA in PCR for diagnosis of PJI. Clinical samples were tested by PCR with and without prior treatment with PMA and compared to conventional microbiological culture. The PCR assay included genus-specific primers for staphylococci and enterococci and species-specific primers for Cutibacterium acnes. The validated conditions of PMA treatment used in this study were 20 μM concentration and 5 and 10 min of dark incubation and photo-activation respectively. 202 periprosthetic tissues and explanted prostheses from 60 episodes in 58 patients undergoing revision arthroplasties for either PJI or non-infective causes were tested, by culture, PCR, and PMA-PCR. 14 of the 60 episodes satisfied the Musculoskeletal Infection Society (MSIS) criteria for PJI and 46 did not. Sensitivity of culture, PCR, and PMA-PCR were 50%, 71%, and 79% respectively. Specificities were 98%, 72%, and 89% respectively. All figures were calculated for episodes rather than samples. PMA-PCR enhanced both the specificity and the sensitivity of PCR. It has the potential to detect residual bacterial viability prior to reimplantation in the two-stage revision for PJI.

Protective effect of polysaccharides from Pholiota nameko on Lactobacillus casei ATCC 334 subjected to freeze-drying

This paper evaluated the benefit of the polysaccharides from Pholiota Nameko (PNPS-1) as a cryoprotectant for maintaining the viability of Lactobacillus casei ATCC 334 during freeze-drying and subsequent storage. The data showed that all the freeze-dried L. casei ATCC 334 samples added with PNPS-1 conformed to the three evaluation indicators of residual viability (>107 CFU/g), moisture content (<7% wet basis) and water activity (<0.6), which were usually used as the quality standards of the freeze-dried powdered starter products. At the same adding concentration of 0.04 mg/ml, PNPS-1 showed only a little weaker than skim milk in the protection for L. casei ATCC 334. However, the combined cryoprotectants contained 0.04 or 0.02 mg/ml PNPS-1 were significantly higher than 0.04 mg/ml skim milk. PNPS-1 could improve the membrane fluidity and membrane integrity compared with the blank control sample. In addition, acid tolerance, bile tolerance and heat tolerance of the freeze-dried cells were all ameliorated by the PNPS-1 protection. This study revealed the novel application of PNPS-1 as a new cryoprotective agent for the freeze-dried L. casei ATCC 334 preservation.

Dual Rinse® HEDP increases the surface tension of NaOCl but may increase its dentin disinfection efficacy.

The aim of this study was to evaluate the surface tension and the antimicrobial activity in infected dentin of a NaOCl solution combined with an etidronate powder (Dual Rinse® HEDP), compared to pure NaOCl and the classic NaOCl + EDTA irrigating sequence, respectively. The surface tension of three irrigants was measured by Wilhelmy technique. To evaluate the antimicrobial activity of the solutions, 26 human teeth were contaminated for 5days with E. faecalis. After bacterial contamination, ten samples were irrigated with NaOCl followed by EDTA, another ten with NaOCl/Dual Rinse® HEDP, and four were used as positive controls. Two specimens not contaminated were used as negative controls. After live/dead BacLight staining, samples were examined by CLSM for analyzing % of residual live and dead cells. Comparison of bacterial viability between and within groups was performed using the Mann-Whitney test for independent samples and the Wilcoxon signed-rank test, respectively. The mean surface tension of EDTA was significantly lower than that of the other irrigants tested (p < 0.001). Conversely, the surface tension of NaOCl/Dual Rinse® HEDP solution was significantly higher than that of all the other solutions (p < 0.001). Residual bacterial viability in the NaOCl/Dual Rinse® HEDP (1.71%) was significantly lower (p = 0.019) than in the NaOCl + EDTA group (3.77%). All of the experimental groups showed significantly lower proportion of viable bacterial cells than the positive control group (p < 0.01). Clinical relevance adding etidronate to NaOCl increases its antimicrobial effect in dentinal tubules even though increases its surface tension.

Влияние режимов сублимационного высушивания на показатели качества вакцины чумной живой

The optimization of the drying schedule has been carried out to improve the quality indicators of the live plague vaccine. Based on the data obtained on the eutectic point of the vaccine suspension, the freezing temperature and freezing time were set to -50 °С and 6-7 h, respectively. A pressure of 40 mTorr over the surface of the drying suspension and 20 mTorr during the desorption were shown to be the best conditions for sublimation. The drying tests with different options for the shelf heating rate, vacuum depth and duration of intermediate temperature indicators were carried out to develop the improved freeze-drying mode providing the selection of the most adapted bacteria. A vaccine lyophilized under the developed conditions has low residual moisture (up to 2%) and high viability index that persists over the whole shelf life. lyophilization, sublimation, eutectic, live plague vaccine, residual moisture, viability

Effect of disinfectant formulation and organic soil on the efficacy of oxidizing disinfectants against biofilms

Biofilms that develop on dry surfaces in the healthcare environment have increased tolerance to disinfectants. This study compared the activity of formulated oxidizing disinfectants with products containing active ingredients against Staphylococcus aureus dry-surface biofilm (DSB) alone. DSB was grown in the CDC bioreactor with alternating cycles of hydration and dehydration. Disinfectant efficacy was tested before and after treatment with neutral detergent for 30 s, and in the presence or absence of standardized soil. Biofilms were treated for 5 min with peracetic acid (Surfex and Proxitane), hydrogen peroxide (Oxivir and 6% H2O2 solution) and chlorine (Chlorclean and sodium dichloroisocyanurate tablets). Residual biofilm viability and mass were determined by plate culture and protein assay, respectively. Biofilm viability was reduced by 2.8 log10 for the chlorine-based products and by 2 log10 for Proxitane, but these products failed to kill any biofilm in the presence of soil. In contrast, Surfex completely inactivated biofilm (6.3 log10 reduction in titre) in the presence of soil. H2O2 products had little effect against DSB. Biofilm mass removed in the presence and absence of soil was <30% by chlorine and approximately 65% by Surfex. Detergent treatment prior to disinfection had no effect. The additives in fully formulated disinfectants can act synergistically with active ingredients, and thus increase biofilm killing whilst decreasing the adverse effect of soil. It is suggested that purchasing officers should seek efficacy testing results, and consider whether efficacy testing has been conducted in the presence of biological soil and/or biofilm.

The Role of Diffusion Weighted Magnetic Resonance Imaging and Subtraction Magnetic Resonance imaging in Assessing Treatment Response of Hepatocellular Carcinoma After Transarterial Chemoembolization

Background: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, and has a poor prognosis unless treated. Ablative therapies are promising treatment options for patients who are not eligible for surgery. Monitoring tumor response after transarterial chemoemolization (TACE) procedure is an important task in oncologic imaging. Early favorable response indicates effectiveness of therapy, while early treatment failure identification is also critical in patient management. In such cases further re-treatment will be mandatory. Aim of work: The aim of this study is to evaluate the efficiency of both diffusion weighted images and subtracted dynamic MRI technique in the detection of residual/ recurrent disease after TACE ablation of non resectable hepatocellular carcinoma (HCC) lesions. Accurate judgment of tumor viability will help diagnose the need for further treatment sessions. Patients and Methods: This study included 32 patients having 54 HCC lesions that underwent transarterial chemoembolization procedure over a period of 6 months (2017- 2018) were followed up 1-1.5 months by dynamic MRI. 12 patients of which underwent a second follow up within 3-4 months. Patients’ ages ranged between 59 to 73 years (mean age 53.1); 26 patients were males and 6 were females. All patients had liver cirrhosis related to chronic viral hepatitis. Results: In 1ry response and follow up findings post TACE, where good response was defied as disappearance of any intra tumoral arterial enhancement in treated lesions, residual disease was defined as at least 30% reduction in the sum of diameters of the viable enhancing lesion in the arterial phase, no response was less than 30% reduction in the enhancing lesion diameters. Progressive disease was defined as 20% increase in the sum of diameters of the enhancing lesions in comparison to the target lesion diameter at the start of treatment. Conclusion: MRI is a powerful tool in detection of residual tumor viability, quantifying tumor necrosis and detecting complications after TACE. Imaging protocol should include dynamic study combined with post processing subtraction images for better tissue characterization.

Neoadjuvant photodynamic therapy augments immediate and prolonged oxaliplatin efficacy in metastatic pancreatic cancer organoids.

Effective treatment of advanced metastatic disease remains the primary challenge in the management of inoperable pancreatic cancer. Current therapies such as oxaliplatin (OxPt)-based chemotherapy regimens (FOLFIRINOX) provide modest short-term survival improvements, yet with significant toxicity. Photodynamic therapy (PDT), a light-activated cancer therapy, demonstrated clinical promise for pancreatic cancer treatment and enhances conventional chemotherapies with non-overlapping toxicities. This study investigates the capacity of neoadjuvant PDT using a clinically-approved photosensitizer, benzoporphyrin derivative (BPD, verteporfin), to enhance OxPt efficacy in metastatic pancreatic cancer. Treatment effects were evaluated in organotypic three-dimensional (3D) cultures, clinically representative models that bridge the gap between conventional cell cultures and in vivo models. The temporally-spaced, multiparametric analyses demonstrated a superior efficacy for combined PDT+OxPt compared to each monotherapy alone, which was recapitulated on different organotypic pancreatic cancer cultures. The therapeutic benefit of neoadjuvant PDT to OxPt chemotherapy materialized in a time-dependent manner, reducing residual viable tissue and tumor viability in a manner not achievable with OxPt or PDT alone. These findings emphasize the need for intelligent combination therapies and relevant models to evaluate the temporal kinetics of interactions between mechanistically-distinct treatments and highlight the promise of PDT as a neoadjuvant treatment for disseminated pancreatic cancer.

A Novel H2S-releasing Amino-Bisphosphonate which combines bone anti-catabolic and anabolic functions

Bisphosphonates (BPs) are the first-line treatment of bone loss resulting from various pathological conditions. Due to their high affinity to bone they have been used to develop conjugates with pro-anabolic or anti-catabolic drugs. We recently demontrated that hydrogen sulfide (H2S), promotes osteogenesis and inhibits osteoclast differentiation. Here we developed an innovative molecule, named DM-22, obtained from the combination of alendronate (AL) and the H2S-releasing moiety aryl-isothiocyanate. DM-22 and AL were assayed in vitro in the concentration range 1-33 μM for effects on viability and function of human osteoclasts (h-OCs) and mesenchymal stromal cells (h-MSCs) undergoing osteogenic differentiation. Amperometric measures revealed that DM-22 releases H2S at a slow rate with a thiol-dependent mechanism. DM-22 significantly inhibited h-OCs differentiation and function, maintaining a residual h-OCs viability even at the high dose of 33 μM. Contrary to AL, in h-MSCs DM-22 did not induce cytotoxicity as revealed by LDH assay, significantly stimulated mineralization as measured by Alizarin Red staining and increased mRNA expression of Collagen I as compared to control cultures. In conclusion, DM-22 is a new BP which inhibits h-OCs function and stimulate osteogenic differentiation of h-MSCs, without cytotoxicity. DM-22 is an ideal candidate for a novel family of osteoanabolic drugs.

Mechanisms of Primary Drug Resistance in FGFR1-Amplified Lung Cancer.

Purpose: The 8p12-p11 locus is frequently amplified in squamous cell lung cancer (SQLC); the receptor tyrosine kinase fibroblast growth factor receptor 1 (FGFR1) being one of the most prominent targets of this amplification. Thus, small molecules inhibiting FGFRs have been employed to treat FGFR1-amplified SQLC. However, only about 11% of such FGFR1-amplified tumors respond to single-agent FGFR inhibition and several tumors exhibited insufficient tumor shrinkage, compatible with the existence of drug-resistant tumor cells.Experimental Design: To investigate possible mechanisms of resistance to FGFR inhibition, we studied the lung cancer cell lines DMS114 and H1581. Both cell lines are highly sensitive to three different FGFR inhibitors, but exhibit sustained residual cellular viability under treatment, indicating a subpopulation of existing drug-resistant cells. We isolated these subpopulations by treating the cells with constant high doses of FGFR inhibitors.Results: The FGFR inhibitor-resistant cells were cross-resistant and characterized by sustained MAPK pathway activation. In drug-resistant H1581 cells, we identified NRAS amplification and DUSP6 deletion, leading to MAPK pathway reactivation. Furthermore, we detected subclonal NRAS amplifications in 3 of 20 (15%) primary human FGFR1-amplified SQLC specimens. In contrast, drug-resistant DMS114 cells exhibited transcriptional upregulation of MET that drove MAPK pathway reactivation. As a consequence, we demonstrate that rational combination therapies resensitize resistant cells to treatment with FGFR inhibitors.Conclusions: We provide evidence for the existence of diverse mechanisms of primary drug resistance in FGFR1-amplified lung cancer and provide a rational strategy to improve FGFR inhibitor therapies by combination treatment. Clin Cancer Res; 23(18); 5527-36. ©2017 AACR.

Effect of baking conditions and storage on the viability of Lactobacillus plantarum supplemented to bread

Bread is an interesting non-dairy-based vehicle for probiotics delivery given its daily consumption worldwide. The incorporation of probiotics in bread is challenging due to the high baking temperatures. In this study the influence of various baking conditions and subsequent storage on survival of a model strain Lactobacillus plantarum P8 is systematically investigated. Bread samples with varying dough weight (5, 30, and 60 g) were baked at different temperatures (175, 205, and 235 ○C) for 8 min, and the residual viability of bacteria was determined every 2 min. Under all baking conditions, the viability of probiotics decreased from 109 CFU/g to 104∼5 CFU/g after baking. For specific conditions a difference in bacterial viability between bread crust and crumb was observed, which was explained by the different temperature-moisture history and developed microstructure during baking. Remarkably, during storage bacterial viability increased by 2–3 log to 108 CFU/g in crust and 106 CFU/g in crumb, respectively. The re-growth of probiotics was accompanied by a decrease in pH of the bread and an increase of the total titratable acidity. The results of this work provide valuable experimental data for further modelling and optimization studies, which then could contribute to the development of probiotic bakery products.