Survival Ratio Research Articles

Implementing precision oncology for sarcoma patients: the CCCLMUmolecular tumor board experience

PurposeDue to poor outcomes and limited treatment options, patients with advanced bone and soft tissue sarcomas (BS/STS) may undergo comprehensive molecular profiling of tumor samples to identify possible therapeutic targets. The aim of this study was to determine the impact of routine molecular profiling in the setting of a dedicated precision oncology program in patients with BS/STS in a German large-volume sarcoma center.Methods92 BS/STS patients who received comprehensive genomic profiling (CGP) and were subsequently discussed in our molecular tumor board (MTB) between 2016 and 2022 were included. Patient records were retrospectively reviewed, and the clinical impact of NGS-related findings was analyzed.Results89.1% of patients had received at least one treatment line before NGS testing. At least one molecular alteration was found in 71 patients (82.6%). The most common alterations were mutations in TP53 (23.3% of patients), followed by PIK3CA and MDM2 mutations (9.3% each). Druggable alterations were identified, and treatment recommended in 32 patients (37.2%). Of those patients with actionable alterations, ten patients (31.2%) received personalized treatment and six patients did benefit from molecular-based therapy in terms of a progression-free survival ratio (PFSr) > 1.3.ConclusionOur single-center experience shows an increasing uptake of next-generation sequencing (NGS) and highlights current challenges of implementing precision oncology in the management of patients with BS/STS. A relevant number of patients were diagnosed with clinically actionable alterations. Our results highlight the potential benefit of NGS in patients with rare cancers and currently limited therapeutic options.

Pulmonary hypertension among preterm infants born at 22 through 32 weeks gestation in France: Prevalence, survival, morbidity and management in the EPIPAGE-2 cohort study

ObjectiveTo determine the prevalence, short-term prognosis and pharmacologic management of pulmonary hypertension (PH) among very preterm infants born before 32 weeks gestation (WG). Study designIn the EPIPAGE-2 French national prospective population-based cohort of preterm infants born in 2011, those presenting with PH were identified and prevalence was estimated using multiple imputation. The primary outcome was survival without severe morbidity at discharge and was compared between infants with or without PH after adjusting for confounders, using generalized estimating equations models. Subgroup analysis was performed according to gestational age (GA) groups. ResultsAmong 3383 eligible infants, 3222 were analyzed. The prevalence of PH was 6.0 % (95 % CI, 5.2–6.9), 14.5 % in infants born at 22–27+6 WG vs 2.7 % in infants born at 28–31+6 WG (P < .001). The primary outcome (survival without severe morbidity at discharge) occurred in 30.2 % of infants with PH vs 80.2 % of infants without PH (P < .001). Adjusted incidence rate ratios for survival without severe morbidity among infants with PH were 0.42 (0.32–0.57) and 0.52 (0.39–0.69) in infants born at 22–27+6 weeks gestation and those born at 28–31+6 weeks, respectively. Among infants with PH, 92.2 % (95 % CI, 87.7–95.2) received sedation and/or analgesia, 63.5 % (95 % CI, 56.6–69.9) received inhaled NO and 57.6 % (95 % CI, 50.9–64.0) received hemodynamic treatments. ConclusionIn this population-based cohort of very preterm infants, the prevalence of PH was 6 %. PH was associated with a significant decrease of survival without severe morbidity in this population.

Early Tumor Shrinkage and Depth of Response as Predictors of Survival for Advanced Biliary Tract Cancer: An Exploratory Analysis of JCOG1113.

Recent studies suggest that early tumor shrinkage (ETS) and depth of response (DpR) reflect outcomes of chemotherapy in various cancers. This study evaluated the association of ETS and DpR with clinical outcomes using data from JCOG1113, which demonstrated the non-inferiority of gemcitabine plus S-1 (GS) to gemcitabine plus cisplatin (GC) for chemotherapy-naïve advanced biliary tract cancer. In total, 354 (289 with measurable target lesions) patients enrolled in JCOG1113 were divided into ETS-unachieved and ETS-achieved groups (≥20% tumor reduction at week 6) and DpR-low and DpR-high groups (≥40% maximum shrinkage) until 12 weeks after enrollment. The impact of ETS and DpR on survival outcome was evaluated using the multivariable Cox proportional hazard model. The proportions of patients in the ETS-achieved and DpR-high groups were similar between the 2 treatment arms. The hazard ratios (HRs) of progression-free survival (PFS) and overall survival (OS) for the ETS-achieved group were 0.70 (95% confidence interval (CI), 0.52-0.93) and 0.60 (95%CI, 0.44-0.81), respectively. The HRs of PFS and OS for the DpR-high group were 0.67 (95%CI, 0.48-0.94) and 0.64 (95%CI, 0.46-0.90), respectively. In the subpopulation treatment effect pattern plot analysis, most patients in the ETS-achieved group in the GC arm did not experience disease progression after 12 weeks from the landmark. As on-treatment markers, ETS and DpR were effective tools. ETS was clinically useful, because it can be used to evaluate the outcomes of treatment early at a specific time.

Evaluation of the relevance of the growth modulation index (GMI) from the FFCD 0307 randomized phase III trial comparing the sequence of two chemotherapeutic regimens

Precision medicine trials disrupted the paradigm of randomized controlled trials in large populations. Patient selection may be based on molecular alterations rather than on primary tumor location. In small patient populations, the growth modulation index (GMI) has been developed to evaluate treatment efficacy by using each patient as its own control. The FFCD 0307 randomized phase III trial compared two sequences of chemotherapy in advanced gastric cancer, which represents a unique opportunity to evaluate the relevance of the GMI. In the FFCD 0307 trial, patients with advanced gastric cancer were randomized between two chemotherapy sequences [ECX followed by FOLFIRI at disease progression (arm A) versus FOLFIRI followed by ECX (arm B)]. GMI was defined as the ratio of the progression-free survival on second treatment (PFS2) to the time to progression on first treatment (TTP1). Sequence benefit was defined as a GMI exceeding 1.3 (GMI-high). GMI was correlated with overall survival (OS). OS1 and OS2 were measured from first randomization and second-line failure to death. Four hundred and sixteen patients were randomized (209 in arm A, 207 in arm B). One hundred and seventy-five patients (42%) received the two sequences and were assessable for GMI (97 in arm A, 79 in arm B). The median GMI was higher in arm A than in arm B (0.62 versus 0.47, P= 0.04). Patients with a high GMI had a longer OS1 (median 14.9 versus 11.5 months, NS). Median OS2 was doubled in the GMI-high group (3.4 versus 1.6 months, NS). GMI analyses suggest that ECX followed by FOLFIRI might represent a better therapeutic strategy than FOLFIRI followed by ECX. High GMI was associated with prolonged survival.

Nineteen-year, real-world experience of first-line combination chemotherapy in patients with metastatic colorectal cancer: a propensity score analysis from southern Thailand.

Combination fluoropyrimidine-based chemotherapy is the standard first-line treatment for metastatic colorectal cancer (CRC). We performed a propensity score (PS)-based analysis to report our real-world experience with long-term follow-up of this regimen for metastatic CRC. In this retrospective study, 170 patients with newly diagnosed metastatic CRC treated with first-line combination chemotherapy between January 2003 and March 2021 were identified. Cox proportional hazards regression analysis and PS-based approaches with the logistic regression model were adopted, and the results were compared. The hazard ratio for overall survival (OS) in the oxaliplatin- and irinotecan-based groups was 0.79 (95% confidence interval = 0.56-1.11), and the median OS times in these groups were 16.8 and 13.0 months, respectively. The median time to progression (TTP) for these regimens were 9.0 and 8.9 months, respectively. The objective response rates for the oxaliplatin- and irinotecan-based groups were 42.7% and 34.6%, respectively. OS and TTP did not differ between these regimens in all PS matching models. First-line treatment using fluoropyrimidine-based chemotherapy regimens in combination with oxaliplatin or irinotecan in patients with metastatic CRC provided comparable efficacy and tolerable toxicity profiles in a real-world setting with long-term follow-up.

Optimal examined lymph node number for accurate staging and long-term survival in rectal cancer: a population-based study.

Although the recommended minimal examined lymph node (ELN) number in rectal cancer (RC) is 12, this standard remains controversial because of insufficient evidence. We aimed to refine this definition by quantifying the relationship between ELN number, stage migration and long-term survival in RC. Data from a Chinese multi-institutional registry (2009-2018) and the Surveillance, Epidemiology, and End Results (SEER) database (2008-2017) on stages I-III resected RC were analysed to determine the relationship between ELN count, stage migration, and overall survival (OS) using multivariable models. The series of odds ratios (ORs) for negative-to-positive node stage migration and hazard ratios (HRs) for survival with more ELNs were fitted using a Locally Weighted Scatterplot Smoothing (LOWESS) smoother, and structural breakpoints were determined using the Chow test. The relationship between ELN and survival was evaluated on a continuous scale using restricted cubic splines (RCS). The distribution of ELN count between the Chinese registry ( n =7694) and SEER database ( n =21332) was similar. With increasing ELN count, both cohorts exhibited significant proportional increases from node-negative to node-positive disease (SEER, OR, 1.012, P <0.001; Chinese registry, OR, 1.016, P =0.014) and serial improvements in OS (SEER: HR, 0.982; Chinese registry: HR, 0.975; both P <0.001) after controlling for confounders. Cut-point analysis showed an optimal threshold ELN count of 15, which was validated in the two cohorts, with the ability to properly discriminate probabilities of survival. A higher ELN count is associated with more precise nodal staging and better survival. Our results robustly conclude that 15 ELNs are the optimal cut-off point for evaluating the quality of lymph node examination and stratification of prognosis.

Effect of water temperature of culture medium on the sex ratio and survival rate of fighting fish Betta dennisyongi larvae

Fighting fish Betta dennisyongi is a popular ornamental freshwater fish in Indonesia. Male fighting fish are preferred because the colour pattern and shape of the fins are more attractive so they are more expensive. This study aims to determine the optimum temperature of rearing media water on the male sex ratio and survival rate of fighting fish larvae. A completely randomized design method with three replications was used in this study. The tested rearing temperature water was: 26°C, 28°C, 30°C, and 32°C. Fish were kept for 40 days. The results of the ANOVA test showed that larval rearing temperature affected significantly the ratio of male and female fish larvae survival (P<0.05). The highest percentage of the male sex was found at a temperature of 32 °C (93.33%), this value was significantly different from other temperature treatments. Meanwhile, the highest larval survival was obtained at a rearing temperature of 28 °C (80.00%), where this value was significantly different from other treatments. Therefore, with economic considerations, it is concluded that the optimal larval rearing media temperature is 28 °C.

Green Synthesis of Novel Antimicrobial ZnO Nanorods

Zinc oxide (ZnO) nanoparticles (NPs) are increasingly used in the diagnosis and prevention of various human infections and diseases. The multi-functional nature of these nanoparticles together with characteristics such as low toxicity and biodegradability, allow their use in a slew of products and applications ranging from biomedical, food and supplement industries, therapeutics and biosensors. ZnO exhibits biomimetic properties enabling biomedical applications, including use as alternatives to pre-existing antibiotics. This study discusses a simple, cost-effective and environmentally sustainable technique for the synthesis of crystalline ZnO nanorods from egg-white or albumin. Single phase nature of the of the engineered nanocrystals was determined using X-ray diffraction (XRD) and selected area electron diffraction (SAED) technique. The morphology of the ZnO nanorods was studied using transmission electron microscopy (TEM) and field emission scanning electron microscopy (FESEM). The diameter of synthesized ZnO nanorods was determined to be in the range of 20–30 nm. The crystal structure of wurtzite ZnO was discovered by the use of Raman, FTIR, while the surface area (12.8 m2/g) was analyzed by using Brunauer-Emmett-Teller (BET) surface area study. Both gram-positive and gram-negative bacterial strains were used to test the ZnO nanorods’ antibacterial abilities without the need of artificial UV activation. Findings showed that different disease-causing bacteria present in community and hospital settings respond differently to ZnO nanorods’ antibacterial activity. When the concentration of nanorods powder increased, the ratio of bacterial survival dropped, showing an increase in antibacterial activity. The gram-negative strain Pseudomonas aeruginosa was shown to have the maximum antibacterial activity of the ZnO nanorods as compared to other gram-negative and positive bacteria. Eco-friendly and green synthesis of ZnO NPs produce vital multifunctional nanomaterials which possess promising antibacterial properties with more extensive studies may develop drugs for multidrug resistance bacteria.

Association of B-Type Natriuretic Peptide Level with Clinical Outcome in Out-of-Hospital Cardiac Arrest in Emergency Department Patients.

B-type natriuretic peptide (BNP) is used for outcome assessment of various diseases. We designed this study to investigate whether BNP, which has been proven useful in the risk stratification of sudden cardiac arrest (SCA) of cardiac etiology, can also prove to be a valuable prognostic tool for SCA also included with non-cardiac etiology. In this study, we aim to investigate the relationship between measured BNP levels and clinical outcomes in SCA, regardless of the cause of SCA. This retrospective multicenter observational study was performed in two tertiary university hospitals and one general hospital between January 2015 and December 2020. The total number of SCA patients was 1625. The patients with out-of-hospital cardiac arrest over 19 years old and acquired laboratory data, including BNP at emergency department (ED) arrival, were included. BNP was measured during advanced Cardiovascular Life Support (ACLS). The exclusion criteria were age under 18 years, traumatic arrest, and without BNP. The median BNP was 171.8 (range; 5-5000) pg/mL in the return of Spontaneous Circulation (ROSC), higher than No-ROSC (p = 0.007). The median BNP concentration was 99.7 (range; 5-3040.68) pg/mL in the survival to discharge, which was significantly lower than the death group (p = 0.012). The odds ratio of survival to discharge decreased proportionally to the BNP level. The odds ratio of neurologic outcome was not correlated with the BNP level. In patients with SCA of all origins, low BNP concentration measured during ACLS correlated with an increased ratio of survival to discharge. However, BNP measured during ACLS was not found to be an independent factor.

Outcomes for Colorectal Cancer Cases With Peritoneal Metastases Treated With Cytoreductive Surgery With or Without Hyperthermic Intraperitoneal Chemotherapy: A Comparative Analysis of Survival Between Peritoneal Carcinomatosis Scores - PCI in a Single Centre.

Peritoneal cancer index (PCI) has been a strong indicator of prognosis for patients receiving cytoreductive surgery. The aim of this single institution study was to compare the survival of peritoneal carcinoma cases treated with cytoreduction surgery arising from colorectal cancer grouped by PCI scores. A retrospective study of a prospective dataset maintained from 2000 till September 2022, for peritoneal metastases from colorectal cancers was carried out. Of the total of 1,625 peritoneal metastases cases, 415 were identified with colorectal cancer and considered for analysis. Survival was followed for 60 months since the index-peritonectomy for cases in this study. Hazard ratio for 5-year survival using the Cox regression analysis over time (t) with a Log-rank (Mantel-Cox) test for significance between the groups indicated, <15 vs. 15 (HR=2.121, p=0.0338), <15 vs. 16-20 (HR=2.748, p<0.0001), <15 vs. >20 (HR=3.158, p<0.0001), 15 vs. 16-20 (HR=1.262, p=0.5658), 15 vs. >20 (HR=1.566, p=0.2771) and for PCI category 16-20 vs. >20 (HR=1.204, p=0.5355) for survival. Median survival for the categories of PCI <15, PCI-15, PCI-16-20, and PCI >15 was 43.967 (95%CI=28.31-59.63), 20.67 (95%CI=5.01-36.33), 19.50 (95%CI=3.84-35.16), and 14.30 (95%CI=1.36-29.96), respectively. A correlation of PCI with survival was confirmed in this study reinforcing the need for assessment of PCI at surgery to help prognostication. Detecting synchronous peritoneal metastases early and prompt treatment can help prevent recurrence and increase survival.

Design of Cinnamaldehyde- and Gentamicin-Loaded Double-Layer Corneal Nanofiber Patches with Antibiofilm and Antimicrobial Effects.

In this study, two-layer poly(vinyl alcohol)/gelatin (PVA/GEL) nanofiber patches containing cinnamaldehyde (CA) in the first layer and gentamicin (GEN) in the second layer were produced by the electrospinning method. The morphology, chemical structures, and thermal temperatures of the produced pure (PVA/GEL), CA-loaded (PVA/GEL/CA), GEN-loaded (PVA/GEL/GEN), and combined drug-loaded (PVA/GEL/CA/GEN) nanofiber patches were determined by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy, and differential scanning calorimetry, respectively. Their mechanical properties, swelling and degradation behavior, and drug release kinetics were investigated. SEM images showed that both drug-free and drug-loaded nanofiber patches possess smooth and monodisperse structures, and nanofiber size increase occurred as the amount of drug increased. The tensile test results showed that the mechanical strength decreased as the drug was loaded. According to the drug release results, CA release ended at the 96th hour, while GEN release continued until the 264th hour. The antibacterial and antibiofilm activities of PVA/GEL, PVA/GEL/CA, PVA/GEL/GEN, and PVA/GEL/CA/GEN nanofiber patches against Pseudomonas aeruginosa and Staphylococcus aureus were evaluated. Results showed that PVA/GEL/GEN and PVA/GEL/CA/GEN nanofiber patches have excellent antibacterial and antibiofilm activities. Moreover, all materials were biocompatible, with no cytotoxic effects in the mammalian cell model for 8 days. PVA/GEL/GEN nanofiber patches were the most promising material for a high cell survival ratio, which was confirmed by SEM images. This research aims to develop an alternative method to stop and treat the rapid progression of bacterial keratitis.

Generation of a human haploid neural stem cell line for genome-wide genetic screening.

Haploid embryonic stem cells (haESCs) have been established in many species. Differentiated haploid cell line types in mammals are lacking due to spontaneous diploidization during differentiation that compromises lineage-specific screens. To derive human haploid neural stem cells (haNSCs) to carry out lineage-specific screens. Human haNSCs were differentiated from human extended haESCs with the help of Y27632 (ROCK signaling pathway inhibitor) and a series of cytokines to reduce diploidization. Neuronal differentiation of haNSCs was performed to examine their neural differentiation potency. Global gene expression analysis was con-ducted to compare haNSCs with diploid NSCs and haESCs. Fluorescence activated cell sorting was performed to assess the diploidization rate of extended haESCs and haNSCs. Genetic manipulation and screening were utilized to evaluate the significance of human haNSCs as genetic screening tools. Human haESCs in extended pluripotent culture medium showed more compact and smaller colonies, a higher efficiency in neural differentiation, a higher cell survival ratio and higher stability in haploidy maintenance. These characteristics effectively facilitated the derivation of human haNSCs. These human haNSCs can be generated by differentiation and maintain haploidy and multipotency to neurons and glia in the long term in vitro. After PiggyBac transfection, there were multiple insertion sites in the human haNSCs' genome, and the insertion sites were evenly spread across all chromosomes. In addition, after the cells were treated with manganese, we were able to generate a list of manganese-induced toxicity genes, demonstrating their utility as genetic screening tools. This is the first report of a generated human haploid somatic cell line with a complete genome, proliferative ability and neural differentiation potential that provides cell resources for recessive inheritance and drug targeted screening.

The Restrictive Red Blood Cell Transfusion Strategy for Critically Injured Patients (RESTRIC) trial: a cluster-randomized, crossover, non-inferiority multicenter trial of restrictive transfusion in trauma.

The efficacies of fresh frozen plasma and coagulation factor transfusion have been widely evaluated in trauma-induced coagulopathy management during the acute post-injury phase. However, the efficacy of red blood cell transfusion has not been adequately investigated in patients with severe trauma, and the optimal hemoglobin target level during the acute post-injury and resuscitation phases remains unclear. Therefore, this study aimed to examine whether a restrictive transfusion strategy was clinically non-inferior to a liberal transfusion strategy during the acute post-injury phase. This cluster-randomized, crossover, non-inferiority multicenter trial was conducted at 22 tertiary emergency medical institutions in Japan and included adult patients with severe trauma at risk of major bleeding. The institutions were allocated a restrictive or liberal transfusion strategy (target hemoglobin levels: 7-9 or 10-12g/dL, respectively). The strategies were applied to patients immediately after arrival at the emergency department. The primary outcome was 28-day survival after arrival at the emergency department. Secondary outcomes included transfusion volume, complication rates, and event-free days. The non-inferiority margin was set at 3%. The 28-day survival rates of patients in the restrictive (n = 216) and liberal (n = 195) strategy groups were 92.1% and 91.3%, respectively. The adjusted odds ratio for 28-day survival in the restrictive versus liberal strategy group was 1.02 (95% confidence interval: 0.49-2.13). Significant non-inferiority was not observed. Transfusion volumes and hemoglobin levels were lower in the restrictive strategy group than in the liberal strategy group. No between-group differences were noted in complication rates or event-free days. Although non-inferiority of the restrictive versus liberal transfusion strategy for 28-day survival was not statistically significant, the mortality and complication rates were similar between the groups. The restrictive transfusion strategy results in a lower transfusion volume. umin.ac.jp/ctr: UMIN000034405, registration date: 8 October 2018.

Selection of optimal quantile protein biomarkers based on cell-level immunohistochemistry data

BackgroundProtein biomarkers of cancer progression and response to therapy are increasingly important for improving personalized medicine. Advanced quantitative pathology platforms enable measurement of protein expression in tissues at the single-cell level. However, this rich quantitative cell-by-cell biomarker information is most often not exploited. Instead, it is reduced to a single mean across the cells of interest or converted into a simple proportion of binary biomarker-positive or -negative cells.ResultsWe investigated the utility of retaining all quantitative information at the single-cell level by considering the values of the quantile function (inverse of the cumulative distribution function) estimated from a sample of cell signal intensity levels in a tumor tissue. An algorithm was developed for selecting optimal cutoffs for dichotomizing cell signal intensity distribution quantiles as predictors of continuous, categorical or survival outcomes. The proposed algorithm was used to select optimal quantile biomarkers of breast cancer progression based on cancer cells’ cell signal intensity levels of nuclear protein Ki-67, Proliferating cell nuclear antigen, Programmed cell death 1 ligand 2, and Progesterone receptor. The performance of the resulting optimal quantile biomarkers was validated and compared to the standard cancer compartment mean signal intensity markers using an independent external validation cohort. For Ki-67, the optimal quantile biomarker was also compared to established biomarkers based on percentages of Ki67-positive cells. For proteins significantly associated with PFS in the external validation cohort, the optimal quantile biomarkers yielded either larger or similar effect size (hazard ratio for progression-free survival) as compared to cancer compartment mean signal intensity biomarkers.ConclusionThe optimal quantile protein biomarkers yield generally improved prognostic value as compared to the standard protein expression markers. The proposed methodology has a broad application to single-cell data from genomics, transcriptomics, proteomics, or metabolomics studies at the single cell level.

Targeting the epithelial-mesenchymal transition (EMT) pathway with combination of Wnt inhibitor and chalcone complexes in lung cancer cells.

Non-small cell lung cancer (NSCLC) is the most common type of the lung cancer. Despite development in treatment options in NSCLC, the overall survival ratios is still poor due to epithelial and mesenchymal transition (EMT) feature and associated metastasis event. Thereby there is a need to develop strategy to increase antitumor response against the NSCLC cells by targeting EMT pathway with combination drugs. Niclosamide and chalcone complexes are both affect cancer cell signaling pathways and therefore inhibit the EMT pathway. In this study, it was aimed to increase antitumor response and suppress EMT pathway in NSCLC cells by combining niclosamide and chalcone complexes. SRB cell viability assay was performed to investigate the anticancer activity of drugs. The drugs were tested on both NSCLC cells (A549 and H1299) and normal lung bronchial cells (BEAS-2B). Then the two drugs were combined and their effects on cancer cells were evaluated. Fluorescence imaging and enzyme-linked immunosorbent assay were performed on treated cells to observe the cell death manner. Wound healing assay, real-time quantitative polymerase chain reaction, and western blot analysis were performed to measure EMT pathway activity. Our results showed that niclosamide and chalcone complexes combination kill cancer cells more than normal lung bronchial cells. Compared to single drug administration, the combination of both drugs killed NSCLC cells more effectively by increasing apoptotic activity. In addition, the combination of niclosamide and chalcone complexes decreased multidrug resistance and EMT activity by lowering their gene expressions and protein levels. These results showed that niclosamide and chalcone complexes combination could be a new drug combination for the treatment of NSCLC.

Establishment of picodroplet-based co-culture system to improve erythritol production in Yarrowia lipolytica

Fluorescence-activated droplet sorting (FADS) combined with transcription factor (TF)-based biosensors allows for the high-throughput isolation of clones that overproduce extracellular metabolites. Here, we developed a FADS-TF coculture pipeline for improving erythritol production in Yarrowia lipolytica. The pipeline comprises three droplet operation steps: the generation and incubation of droplets that encapsulate yeast mutants with single-cell resolution, the pico-injection of fluorescence-based erythritol-biosensing Escherichia coli, and the sorting of the highest fluorescence droplets with an approximately 1 ‰ survival ratio. With the picodroplet-based workflow, we initially carried out a two-stage temperature and pH control strategy on the droplets to separate the erythritol production and detection processes. We then eliminated the background expression of biosensors by utilizing pH-sensitive erythromycin, finally establishing a complete version of the coculture screening system. The optimal strain, Yarrowia lipolytica S4-9, contributed 231.2 g/L erythritol after a 114-h fermentation and was successfully screened from the fourth round of the mutant library. Compared to the parent, the erythritol yield and production rate of Yarrowia lipolytica S4-9 increased by 16.97 % and 26.09 %, respectively, in a 5-L bioreactor. Our results indicated that FADS-TFs have great potential for the high-throughput screening of extracellular products as additional biosensors become available.

First-line immunotherapy combinations for recurrent or metastatic nasopharyngeal carcinoma: An updated network meta-analysis and cost-effectiveness analysis.

Recently updated results of randomized clinical trials (RCTs) have confirmed that toripalimab, camrelizumab, and tislelizumab plus chemotherapy (TOGP, CAGP, and TIGP) significantly prolonged survival compared to placebo plus chemotherapy (PLGP) in the first-line treatment for recurrent or metastatic nasopharyngeal carcinoma (R/M-NPC). However, the high cost of immunotherapies imposes a huge financial burden on patients and health care systems. RCTs estimating immunotherapies for R/M-NPC were searched. A Bayesian network meta-analysis (NMA) was carried out; the main outcomes were hazard ratios (HRs) of overall survival (OS) and progression-free survival (PFS). The cost and efficacy of four first-line therapies were evaluated using the Markov model. The main outcome in the cost-effectiveness analysis (CEA) was incremental cost-utility ratios (ICURs). The model robustness was assessed by one-way, three-way, and probabilistic sensitivity analyses. Three RCTs (JUPITER-02, CAPTAIN-1st, and RATIONALE-309) involving 815 patients were included in the NMA. Compared with PLGP, chemo-immunotherapies have significantly longer PFS and OS. Compared to the PLGP group, TOGP, CAGP, and TIGP groups resulted in additional costs of $48 339, $22 900, and $23 162, with additional 1.89, 0.73, and 0.960 QALYs, respectively, leading to the ICURs of $25 576/QALY, $31 370/QALY, and $31 729/QALY. Pairwise comparisons showed TOGP was the most cost-effective option among chemo-immunotherapy groups. From the Chinese payers' perspective, first-line immunotherapy combination therapies provided significant survival and cost-effectiveness superiority over chemotherapy alone for patients with R/M-NPC at the WTP of $38 029/QALY. Among the three chemo-immunotherapy groups, TOGP was the most cost-effective option.

Stem Cell Therapy in Critical Limb Ischemia.

Critical limb ischemia (CLI), a serious outcome of peripheral artery disease, is frequently associated with morbid outcomes. The available treatment modalities do not provide satisfactory results, leading to marked morbidities such as joint contracture and amputations, resulting in a high economic burden. The peripheral vascular disease tends to cause more morbidity in patients with diabetes and atherosclerosis, given the pre-existing compromised perfusion of medium and small vessels in diabetic patients. With surgical procedures, the chance of vascular compromise further increases, inducing a significantly greater rate of amputation. Hence, the need for nonsurgical treatment modalities such as stem cell therapy (SCT), which promotes angiogenesis, is warranted. In CLI, SCT acts through neovascularization and the development of collateral arteries, which increases blood supply to the soft tissues of the ischemic limb, providing satisfactory outcomes. An electronic database search was performed in PubMed, SCOPUS, EMBASE, and ScienceDirect to identify published clinical trial data, research studies, and review articles on stem cell therapy in critical limb ischemia. The search resulted in a total of 2391 results. Duplicate articles screening resulted in 565 articles. In-depth screening of abstracts and research titles excluded 520 articles, yielding 45 articles suitable for full-text review. On review of full text, articles with overlapping and similar results were filtered, ending in 25 articles. SCT promotes arteriogenesis, and bone marrow-derived mesenchymal stromal cells produce significant effects like reduced morbidity, improved amputation-free survival (AFS ) rate, and improved distal perfusion even in "no-option" CLI patients. SCT is a promising treatment modality for CLI patients, even in those in whom endovascular and revascularization procedures are impossible. SCT assures a prolonged AFS rate, improved distal perfusion, improved walking distances, reduced amputation rates, and increased survival ratio, and is well-tolerated.

Modeling survival curves of Anisakis L3 after isothermal heat treatments at lethal temperatures

Even though anisakiasis is considered, nowadays, a significant threat to public health all over the world, no attempt has been made up to date to mathematically describe the thermal susceptibility of Anisakis larvae in the third stage (L3). To fill this gap, in this paper, more than 10,000 free (non-encysted) Anisakis L3 were individually heat treated in a thermal cycler at temperatures between 44 °C and 61 °C for different exposure times. After heat exposition, viability was assessed in each larva, survival curves at isothermal conditions were derived, and the effectiveness of four kinetic models (fundamental kinetic model, Mafart model, and probit and logit models) in describing these curves was tested. Evaluation of larvae viability after heat exposition revealed sigmoidal survival curves that increased their steepness with temperature. Of the four models tested, the Mafart model was the one that best fitted the data only differing from the observed survival ratios by 0.12 units on average. Validation experiments performed at temperatures different to those used to create the model corroborated its predictive capacity. Future efforts should be focused in predicting larvae viability at non-isothermal conditions as those occurring during fish cooking.

Cost-effectiveness of encorafenib with binimetinib in unresectable or metastatic BRAF-mutant melanoma.

The objective of this study was to determine the cost-effectiveness of encorafenib with binimetinib (EncoBini) as compared to other targeted double combination therapies, namely dabrafenib with trametinib (DabraTrame) and vemurafenib with cobimetinib (VemuCobi), for the treatment of BRAF V600-mutant unresectable or metastatic melanoma (MM) from the French payer perspective. A partitioned survival model was developed considering a lifetime horizon. The model structure simulated the clinical pathway of patients with BRAF V600-mutant MM. Clinical effectiveness and safety inputs were sourced from the COLUMBUS trial, a network meta-analysis and published literature. Costs, resource use, and the quality of life inputs were obtained from the literature and appropriate French sources. Over a lifetime horizon, EncoBini was associated, on average, with reduced costs and increased quality adjusted life years (QALYs), dominating both targeted double combination therapies. For a willingness-to-pay threshold of €90,000 per QALY, the probability of EncoBini being cost-effective against either comparator remained above 80%. The most influential model parameters were the hazard ratios for the overall survival of EncoBini vs DabraTrame and VemuCobi, the pre- and post-progression utility values, as well as treatment dosages and the relative dose intensity of all interventions. EncoBini is associated with reduced costs and increased QALYs, dominating other targeted double combination therapies (DabraTrame, VemuCobi) for patients with BRAF V600-mutant MM in France. EncoBini is a highly cost-effective intervention in MM.