Short Doubling Time Research Articles

Effect on Growth Performance and Nutritive Value of Cultivated Azolla filiculoides As An Alternative Feedstuff for Ruminant.

Azolla filiculoides is a tiny, free-floating aquatic fern and has a potential alternative protein and fibre source for ruminants, was investigated for its cultivation optimisation and feedstuff suitability. Study 1 was conducted to investigate the influence of different fertiliser types (control, broiler manure, sheep manure, cow manure) and concentrations (0.25 g/L-1.25 g/L) on the growth performance (fresh weight, doubling time, relative growth rate) and nutrient composition (dry matter, ash, crude protein, crude fibre, crude fat) of A. filiculoides. The optimised type of fertiliser and concentration in Study 1 were further adopted in Study 2 to evaluate the effect of different fertiliser processing methods on the growth performance, nutritive value and in vitro rumen digestibility of A. filiculoides upon cultivation. The findings in Study 1 showed that cultivation of A. filiculoides using sheep manure at the concentration of 1.00 g/L is the best resulted in the shortest doubling time (3 to 5 days) and produced fresh weight (FW), relative growth rate (RGR), crude protein (CP) and crude fibre (CF) at 132.2 g/m2, 0.32 g/g/day, 21.2% DM-1 and 14.4% DM-1, respectively. Furthermore, unprocessed sheep manure (T3) exhibited superior (p < 0.05) fresh weight, relative growth rate, nutrient composition and fibre components compared to the burned manure treatment (T2). In vitro digestibility analysis discovered that T3 achieved a 24-hour accumulated gas production of 86.9 mL DM-1, with in vitro dry matter digestibility (IVDMD), in vitro organic matter digestibility (IVOMD) and metabolisable energy (ME) of 82.9%, 43.7% and 5.8 MJ/kg DM, respectively. These findings suggest that Azolla filiculoides cultivation can be economically optimised using 1.00 g/L unprocessed sheep manure (fresh manure), potentially serving as a self-produced, nutritious feedstuff for ruminants.

Pollution control and biodiesel production with microalgae: new perspectives on the use of flat panel photobioreactors regarding variation in volume application rate.

In the present study, the microalga Arthrospira platensis DHR 20 was cultivated in vertical flat-plate photobioreactors (FPBRs) to bioremediate anaerobically digested cattle wastewater (ACWW) and used as a growth substrate. The final objective was to evaluate the properties of the oil extracted from this biomass to determine its potential for biodiesel production. The process was divided into five phases, varying the volume of the applied substrate: 1 L (Phase I), 5 L (Phase II), 10 L (Phase III), 15 L (Phase IV), and 20 L (Phase V). Dry biomass reached a maximum of 5.7g L-1, and productivity peaked at 0.74g L-1d-1. The highest rate of CO2 biofixation was 1213.5mg L-1day-1, showing good potential for purifying the air. The highest specific maximum growth rate (μmax) and the shortest doubling time (Dt) were found during Phase I. The removal of pollutants and nutrients during the experimental phases ranged from 65.8% to 87.1% for chemical oxygen demand (COD), 82.2% to 85.8% for total organic carbon (TOC), 91% to 99% for phosphate (PO43-), 62.5% to 93% for nitrate (NO3-), 90.4% to 99.7% for ammoniacal nitrogen (NH4+), and 86.5% to 98.5% for total nitrogen (TN). The highest lipid production recorded was 0.172g L-1day-1. The average cetane number recorded in Phase IV of 51 suggests that the fuel will ignite efficiently and consistently, providing smooth operation and potentially reducing pollutant emissions. The analysis of fatty acids revealed that the produced biodiesel has the potential to be used as an additive for other low-explosive biocombustibles, representing an innovative and sustainable approach that simultaneously offers bioremediation and carbon sequestration.

Real-world outcomes following biochemical recurrence after definitive therapy with a short prostate-specific antigen doubling time: potential role of early secondary treatment.

The natural history of biochemical recurrence (BCR) managed with delayed hormonal therapy is well documented by data from Johns Hopkins. However, as many patients receive treatment prior to metastasis, we evaluated the natural history and role of prostate-specific antigen doubling time (PSADT) in a more contemporary cohort of BCR patients with nonmetastatic castration-sensitive prostate cancer (nmCSPC). Patients in the Veterans Health Administration (VHA; 01/01/06 to 06/22/20) with nmCSPC and BCR were divided into rapid ( ≤9 months) and less rapid ( >9 to ≤15 months) PSADT cohorts. Patients with PSADT >15 months were excluded as outcomes, even with delayed treatment, are excellent. Outcomes included time to first antineoplastic therapy after BCR, metastasis, metastasis-free survival (MFS), and overall survival (OS). Cox models adjusted for baseline demographics and clinical characteristics. Overall, 781 patients with BCR were identified (502 rapid; 279 less rapid PSADT). Rapid PSADT was associated with shorter time to first systemic antineoplastic therapy (median 11.4 vs. 28.3 months, adjusted hazard ratio [95% confidence interval] 2.17 [1.83-2.57]), metastasis (102.4 months vs. not reached, 1.79 [1.33-2.40]), MFS (76.1 vs. 106.3 months, 1.73 [1.33-2.24]), and OS (120.5 vs. 140.5 months, 1.76 [1.22-2.54]) versus less rapid PSADT. Most patients with rapid PSADT underwent secondary treatment within 1 year after BCR. More contemporary patients treated with early secondary treatment had better outcomes than historical data from patients who had delayed treatment. Whether these results reflect the benefits of early secondary treatment or overall improvements in prostate cancer outcomes over time requires further study.

Evaluation of the synbiotic effects of Saccharomyces cerevisiae and mushroom extract on the growth performance, digestive enzyme activity, and immune status of zebrafish danio rerio

BackgroundThe quest for candidate probiotics and prebiotics to develop novel synbiotics for sustainable and profitable fish farming remains a major focus for various stakeholders. In this study, we examined the effects of combining two fungal probiotics, Saccharomyces cerevisiae and Aspergillus niger with extracts of Jerusalem artichoke and white button mushroom to develop a synbiotic formulation to improve the growth and health status of zebrafish (Danio rerio). An initial in vitro study determined the most effective synbiotic combination, which was then tested in a 60-day in vivo nutritional trial using zebrafish (80 ± 1.0 mg) as a model animal. Four experimental diets were prepared: a control diet (basal diet), a prebiotic diet with 100% selected mushroom extract, a probiotic diet with 107 CFU of S. cerevisiae/g of diet, and a synbiotic diet with 107 CFU of S. cerevisiae/g of diet and 100% mushroom extract. As readouts, growth performance, survival, digestive enzyme activity and innate immune responses were evaluated.ResultsIn vitro results showed that the S. cerevisiae cultured in a medium containing 100% mushroom extract exhibited the maximum specific growth rate and shortest doubling time. In the in vivo test with zebrafish, feeding them with a synbiotic diet, developed with S. cerevisiae and mushroom extract, led to a significant improvement in the growth performance of zebrafish (P < 0.05). The group of zebrafish fed with the synbiotic diet showed significantly higher levels of digestive enzyme activity and immune responses compared to the control group (P < 0.05).ConclusionTaken together, these results indicated that the combination of S. cerevisiae and mushroom extract forms an effective synbiotic, capable of enhancing growth performance and immune response in zebrafish.

Predictors of Progression to Castration-resistant Prostate Cancer After Radical Prostatectomy in High-risk Prostate Cancer Patients.

To examine the specific time frame and identify associated risk factors from commencement of hormonal therapy to the onset of castration-resistant prostate cancer among patients who have developed biochemical recurrence following radical prostatectomy. We retrospectively reviewed the records of 92 patients who developed biochemical recurrence and received hormonal therapy as initial salvage treatment after radical prostatectomy for high-risk localized prostate cancer from 2005 to 2021. The castration-resistant prostate cancer-free survival rates from the commencement of salvage hormonal therapy were analyzed using log-rank methods. Cox proportional hazard regression was performed to analyze the risk factors associated with acquiring castration resistance. The patients were stratified based on those risk factors. During a median follow-up duration of 57 months, 24 (26.1%) patients developed castration-resistant prostate cancer. The 5- and 10-year castration-resistant prostate cancer-free survival rates were 73.6% and 54.5%, respectively. A multivariate analysis showed that Grade Group of 5 and prostate-specific antigen doubling time at biochemical recurrence of ≤3 months were independent predictors of castration-resistant prostate cancer. The 5-year castration-resistant prostate cancer-free survival rates in the low- and high-risk groups, stratified according to the aforementioned factors, were 85.4% and 47.6%, respectively. Patients in high Grade Group and short prostate-specific antigen doubling time after radical prostatectomy are more likely to develop resistance to salvage hormonal therapy.

Assessing olaparib efficacy in U.S. Veterans with metastatic prostate cancer utilizing a time-indifferent g-rate method ideal for real-world analyses

We aimed to assess real-world efficacy of the PARP inhibitor, olaparib, in US Veterans with metastatic prostate cancer (mPC) by leveraging the national data repository and evaluate a novel approach to assess treatment efficacy in tumors considered rare or harboring rare mutations. Included Veterans had 1) mPC with somatic or germline alterations/mutations in genes involved in homologous recombination repair (HRR), 2) received olaparib monotherapy as well as a novel hormonal therapy/androgen receptor pathway inhibitors (NHT/ARPI), and/or chemotherapy, and 3) estimable rates of tumor growth (g-rate) using PSA values obtained while receiving treatment. Previous work has shown an excellent inverse correlation of g-rate with survival. Using g-rate, we determined tumor doubling time (DT) and DT ratios (DT on olaparib/DT on prior medication). We postulated that a DT ratio≥ 1 was associated with benefit. We identified 139 Veterans, including 42 Black males with tumors harboring mutations/alterations in HRR genes who received olaparib: BRCA2 (50), ATM (32), BRCA1 (10), other mutations (47). 62/139 (45%) of all and 21/42 (50%) of Black Veterans had DT ratios ≥1, including 31, 10, 2, and 19 with BRCA2, ATM, BRCA1, and other mutations, respectively (p=0.006). Median survival with DT ratios ≥1 was superior, being 24.5 vs. 11.4 months for DT ratio <1 (p=0.01, HR 0.50, 95% CI 0.29-0.85). Benefit from olaparib, defined as DT ratio ≥1, was not observed for germline status, starting PSA value, number of prior therapies, or immediate prior therapy. Compared to matched cohorts, tumors in the olaparib cohort had shorter DTs with enzalutamide in first line (367 vs. 884 days; p=0.0043). Using equations indifferent to timing of assessments ideal for real-world efficacy analyses, we showed DT ratio ≥1 representing slower tumor growth on olaparib relative to the prior therapy correlates with improved survival. Olaparib efficacy in Veterans with mPC harboring mutations/alterations in HRR genes emulates clinical trial results. Black men had comparable results. Compared to matched cohorts, in first line, enzalutamide was less efficacious in tumors harboring mutations/alterations in HRR genes. American Society of Clinical Oncology Conquer Cancer Foundation (ASCO CCF), the Blavatnik Family Foundation and the Prostate Cancer Foundation (PCF).

Establishment of porcine embryonic stem cells in simplified serum free media and feeder free expansion

BackgroundThe establishment of stable porcine embryonic stem cells (pESCs) can contribute to basic and biomedical research, including comparative developmental biology, as well as assessing the safety of stem cell-based therapies. Despite these advantages, most pESCs obtained from in vitro blastocysts require complex media and feeder layers, making routine use, genetic modification, and differentiation into specific cell types difficult. We aimed to establish pESCs with a single cell-passage ability, high proliferative potency, and stable in long-term culture from in vitro-derived blastocysts using a simplified serum-free medium.MethodsWe evaluated the establishment efficiency of pESCs from in vitro blastocysts using various basal media (DMEM/F10 (1:1), DMEM/F12, and a-MEM) and factors (FGF2, IWR-1, CHIR99021, and WH-4-023). The pluripotency and self-renewal capacity of the established pESCs were analyzed under feeder or feeder-free conditions. Ultimately, we developed a simplified culture medium (FIW) composed of FGF2, IWR-1, and WH-4-023 under serum-free conditions.ResultsThe pESC-FIW lines were capable of single-cell passaging with short cell doubling times and expressed the pluripotency markers POU5F1, SOX2, and NANOG, as well as cell surface markers SSEA1, SSEA4, and TRA-1-60. pESC-FIW showed a stable proliferation rate and normal karyotype, even after 50 passages. Transcriptome analysis revealed that pESC-FIW were similar to reported pESC maintained in complex media and showed gastrulating epiblast cell characteristics. pESC-FIW were maintained for multiple passages under feeder-free conditions on fibronectin-coated plates using mTeSR™, a commercial medium used for feeder-free culture, exhibiting characteristics similar to those observed under feeder conditions.ConclusionsThese results indicated that inhibition of WNT and SRC was sufficient to establish pESCs capable of single-cell passaging and feeder-free expansion under serum-free conditions. The easy maintenance of pESCs facilitates their application in gene editing technology for agriculture and biomedicine, as well as lineage commitment studies.

Producing recombinant proteins in Vibrio natriegens

The diversity of chemical and structural attributes of proteins makes it inherently difficult to produce a wide range of proteins in a single recombinant protein production system. The nature of the target proteins themselves, along with cost, ease of use, and speed, are typically cited as major factors to consider in production. Despite a wide variety of alternative expression systems, most recombinant proteins for research and therapeutics are produced in a limited number of systems: Escherichia coli, yeast, insect cells, and the mammalian cell lines HEK293 and CHO. Recent interest in Vibrio natriegens as a new bacterial recombinant protein expression host is due in part to its short doubling time of ≤ 10 min but also stems from the promise of compatibility with techniques and genetic systems developed for E. coli. We successfully incorporated V. natriegens as an additional bacterial expression system for recombinant protein production and report improvements to published protocols as well as new protocols that expand the versatility of the system. While not all proteins benefit from production in V. natriegens, we successfully produced several proteins that were difficult or impossible to produce in E. coli. We also show that in some cases, the increased yield is due to higher levels of properly folded protein. Additionally, we were able to adapt our enhanced isotope incorporation methods for use with V. natriegens. Taken together, these observations and improvements allowed production of proteins for structural biology, biochemistry, assay development, and structure-based drug design in V. natriegens that were impossible and/or unaffordable to produce in E. coli.

#848 Influence of nutrition and other factors on preterm newborns’ kidney development

Abstract Background and Aims According to WHO, in 2020, preterm newborns represented around 10% of all newborns in the world. Premature birth is deeply connected with the risk of developing a chronic kidney disease later in life. This is caused by defects in kidney development (primarily the lower number of nephrons) that are often difficult to identify in the complicated clinical picture of premature babies. Our study aims at providing knowledge on the relationship between preterm infant kidney development and morbidities, nutrition, and blood biomarkers. Method Creatinine was measured in 130 extremely preterm newborns (less than 28 weeks of gestation) enrolled in a randomized controlled trial (Mega Donna Mega) in two Swedish neonatal intensive care units. Creatinine was measured in whole blood or serum with the enzymatic method. Individuals with 4 or more serum creatinine measurements (n = 92) were shortlisted for eGFR calculation. eGFR was calculated with the Wilhelm-Bals weight-based equation due to the lack of length measurements. The patients were divided in 3 groups based upon their kidney development status. This was indirectly assessed using the exponential regression curve describing the eGFR trends as model and the doubling time as discriminating parameter. Data about morbidity, treatment, nutrition, blood lipidomics and proteomics were also collected during the MegaDonnaMega trial and has been used to investigate the differences between the three groups. A linear model adjusted for sex and gestational age was used to analyse lipidomics and proteomics data in relation to eGFR. The omics analysis used average data from the first month of age. Data analysis was performed in R, SPSS, SAS and GraphPad statistical programs. Results Subjects were divided in 3 groups from best to worst kidney function development. The parameter used, doubling time, is inversely correlated to eGFR and kidney development. Subjects in group 1 reached the normal at-term birth eGFR (40 ml/min/1.73 m2) at around 50 days of age, whereas the other two groups were delayed (66 days for group 2, 78 for group 3). Subjects in group 2 and 3 were characterized by longer stay at the NICU, lower weight and length in respect to infants in group 1. Subjects in groups 2 and 3 also showed more co-morbidities in respect to group 1. Regarding nutrition, subjects administered with maternal milk had shorter doubling times when compared to those administered with donor or mixed milk. Six serum fatty acids correlated with the eGFR trends: heptadecanoic (FA 17:0) showed a negative correlation, while palmitoleic (16:1n-7), gamma-linolenic (18:3n-6), erucic (22:1n-9) docosapentaenoic (22:5n-6) and tricosanoic (23:0) acid showed positive correlations. 45 serum proteins correlated with the eGFR trends, in particular, inflammatory and proliferative pathways showed an inverse correlation with eGFR. Conclusion We have detected differences in the eGFR trends of preterm newborns that could be traced back to nutrition and other factors. Moreover, we identified several serum fatty acids and proteins that correlated with eGFR and therefore kidney development. The data indicates that postnatal kidney development may be important to consider in the clinical care of preterm newborns.

Rationally designed chromosome fusion does not prevent rapid growth of Vibrio natriegens

DNA replication is essential for the proliferation of all cells. Bacterial chromosomes are replicated bidirectionally from a single origin of replication, with replication proceeding at about 1000 bp per second. For the model organism, Escherichia coli, this translates into a replication time of about 40 min for its 4.6 Mb chromosome. Nevertheless, E. coli can propagate by overlapping replication cycles with a maximum short doubling time of 20 min. The fastest growing bacterium known, Vibrio natriegens, is able to replicate with a generation time of less than 10 min. It has a bipartite genome with chromosome sizes of 3.2 and 1.9 Mb. Is simultaneous replication from two origins a prerequisite for its rapid growth? We fused the two chromosomes of V. natriegens to create a strain carrying one chromosome with a single origin of replication. Compared to the parental, this strain showed no significant deviation in growth rate. This suggests that the split genome is not a prerequisite for rapid growth.

Enhanced production of withaferin A from the hairy root culture of Withania somnifera via synergistic effect of Methyl jasmonate and β-cyclodextrin

Due to low amounts of withanolides produced in some plants and high demand for various applications, their biotechnological production is widely researched. The effects of two explant types (i.e., leaf and stem from the in vitro seedlings of three genotypes of Withania somnifera) and four Rhizobium strains (i.e., LBA 9402, A4, ATCC 15834, and C58C1) to improve hairy root formation efficiency was studied. Furthermore, the combined effects of β-cyclodextrin (β-CD) and methyl jasmonate (MeJA) on withaferin A production after 48 h exposure time was examined. Four hairy roots having the maximum percentage of induced roots and mean number of induced roots to analyze their growth kinetics and identified G3/ATCC/LEAF culture having the maximum specific growth rate (μ = 0.036 day−1) and growth index (GI = 9.18), and the shortest doubling time (Td = 18.82 day) were selected. After 48 h exposure of G3/ATCC/LEAF culture to different elicitation conditions, maximum amounts of withaferin A were produced in samples co-treated with 0.5 mM β-CD + 100 μM MeJA (9.57 mg/g DW) and 5.0 mM β-CD + 100 μM MeJA (17.45 mg/g DW). These outcomes represented a 6.8-fold and 12.5-fold increase, respectively, compared to the control. Similarly, combined β-CD/MeJA elicitation increased gene expression levels of HMGR, SQS, SMT-1, and SDS/CYP710A involved in withanolides biosynthetic pathway, of which just SMT-1 had significant correlation with withaferin A production. These results demonstrated the superiority of G1-leaf explant and ATCC 15834 for hairy root induction, and revealed synergistic effect of MeJA and β-CD on withaferin A production.

Addition of Chidamide to a Sandwich Protocol, Pegaspargase, Gemcitabine and Oxaliplatin Chemotherapy Combined with Radiotherapy, As First-Line Treatment in Early Stage NK/T-Cell Lymphoma: Real World Outcomes in a Retrospective Study

Background: Extranodal NK/T-cell lymphoma, nasal type (ENKTL-NT) is a highly invasive tumor with a short doubling time and poor prognosis. Clinically, it usually involves the upper aerodigestive tract and mainly presents with early-stage. Initially, intensity modulated radiation therapy (IMRT) has been considered the preferred first-line therapy for early-stage ENKTL due to its high response rate. In recent years, more and more studies show that adding chemotherapy, especially asparaginase (ASP)-based regimens, provide survival benefit in early-stage patients. However, the optimal chemotherapy regimen has not been fully defined. Epigenetic regulatory mechanisms related to histone deacetylase (HDAC) have been found to play a role in the evolution and progression of ENKTL. Chidamide, a new HDAC inhibitor, has been confirmed to be effective and tolerable to treat lymphoma. Recently, Xue et al showed that chidamide in combination with cisplatin, etoposide or gemcitabine showed synergistic effect and reversed the chemotherapy resistance in the relapsed/refractory B-cell lymphoma. Here, we investigate the safety and efficacy of adding chidamide to a sandwich protocol, P-GEMOX (pegaspargase/gemcitabine/oxaliplatin) regimen (CP-GEMOX) combined with IMRT as first-line treatment in early-stage ENKTL-NT patients. Methods: We retrospectively reviewed data of early-stage ENKTL-NT patients in the first affiliated hospital of Soochow University from March 2020 to April 2023. A total of 21 patients diagnosed with early-stage (stage Ⅰ or Ⅱ) ENKTL-NT were enrolled and treated with 2 cycles of CP-GEMOX followed by IMRT and 1 or 2 cycles of CP-GEMOX sequentially as consolidation. After consolidation, chidamide was administrated as a maintenance therapy for 1 year. In the induction therapy phase, chidamide was administered orally at a dose of 20 mg twice a week for two weeks, with a one-week discontinuation period; During the maintenance treatment phase, the original dose of chidamide was still maintained and used without pause. The baseline clinical features for the 21 patients were summarized in Table 1. Results: Among 21 patients, 14 were males and 7 were females, and their median age was 57 (range, 41-75) years. Elevated lactate dehydrogenase, elevated β2-microglobulin, stage II disease and primary tumor invasion were present in 4.8% (1/21), 19.0% (4/21), 61.9% (13/21) and 61.9% (13/21) patients. According to the early stage-adjusted nomogram-revised risk index, the majority of patients (76.2%, 16/21) were classified into intermediate- (1-2) and high (≥3) risk groups. The baseline clinical features for the 21 patients were summarized in Table 1. During treatment, all patients were well-tolerated and the most common adverse events were hematological toxicities with neutropenia, thrombocytopenia and anemia. Among non-hematological adverse events, nausea, vomiting and reduced appetite were most common. Other side effects included coagulation dysfunction, skin reaction, diarrhea, serum amylase and alanine aminotransferas /aspartate aminotransferase /bilirubin elevation. No anaphylactic reaction to pegaspargase, hemophagocytic syndrome, pancreatitis or sepsis were observed during chemotherapy. Some patients had mucositis and xerostomia when receiving local radiotherapy. All side effects could be reversed with proper treatment, and there were no therapy related mortality. All patients completed 2 cycles of CP-GEMOX, 19 patients achieved complete remission (CR) and 2 patients achieved partial remission (PR) with a CR rate of 90.5% (19/21) and overall response rate (ORR) of 100.0% (21/21) after 2 cycles of CP-GEMOX. Among the 21 patients, 18 patients completed treatment and CR rate was improved to 100.0% (18/18), giving an ORR of 100% (18/18). After a median follow-up of 25.6(range, 2.6-39) months , all patients were alive except one patient relapsed 20 months after treatment and died of COVID-19 related pneumonia 1 month later. The 2-year progression-free survival and overall survival were all 91.5% (Figure 1). Conclusions: This study demonstrates that sandwich CP-GEMOX chemoradiotherapy is a safe and promising approach to managing patients with early-stage ENKTL-NT. Keywords: Extranodal NK/T-cell lymphoma; chidamide; pegaspargase; gemcitabine; radiation therapy.

Remoción de nitrógeno y fósforo acoplada a la fijación de CO2 de dos microalgas verdes, Chlorella sp. y Quadrigula sp., nativas de la costa de Chiapas, México

Microalgae remove nitrogen and phosphorus from water for their growth and biomass production, wich has several biotechnological applications. This study evaluated the growth, NO3−, NO2−, NH4+, urea, PO43− and total nitrogen removal capacity, as well as the biomass production and the CO2 fixation of two new microalgae isolates, Chlorella sp. and Quadrigula sp., from the coast of Chiapas, México. Chlorella sp. had a higher specific growth rate (0.74/day) and a shorter doubling time (0.92 days) than Quadrigula sp. Removal per unit volume (mg/L) of NO3−, NH4+ and total nitrogen by Chlorella sp. was higher (26.88%, 10.58% and 16.63%, respectively) than removal by Quadrigula sp. On the other hand, daily removal (mg/L/d) of NO3−; NH4+; NO2−; urea and total nitrogen by Chlorella sp. was also higher (88%, 33%, 89%,15% and 50%, respectively) than daily removal by Quadrigula sp. The biomass productivity (mg/L/d) and the CO2 fixation rate (mg/L/d) were similar among both microalgae. The results of this study suggest that Chlorella sp. and Quadrigula sp. have potential for nutrient removal, biomass production and CO2 fixation.

Effective CO2 capture by the fed-batch culture of Chlorella vulgaris

Microalgae are promising candidates for CO2 capture and concomitant biomass production. Chlorella vulgaris was grown in the batch and fed-batch cultures to develop a culture strategy targeting maximum CO2 capture and conversion to biomass. Growth at five different dissolved CO2 (dCO2) concentrations in the feeding media (atmospheric, 1.55, 1.62, 1.75 and 1.88 g L−1) was tested to assess the effect of dCO2 concentration on the growth, biomass productivity, and nutrients removal efficiency of microalgae. Results suggest that fed-batch culture outperformed “standard” batch cultivation with a higher algal growth rate (2.3 times). As expected, the algal growth was limited at low (atmospheric) dCO2 concentrations and inhibited at high levels (1.75 and 1.88 g L−1). C. vulgaris grown with medium containing 1.62 g L−1 dCO2 showed the highest growth rate (0.094 h−1) with the shortest doubling time (7.4 h), maximum biomass productivity, nitrogen and phosphorus uptake rates (222, 7.5 and 1.6 mg L−1 d−1, respectively). By pre-dissolving CO2 into the feeding media and using the fed-batch culture mode, a high CO2 removal efficiency could be achieved.

New methods to increase microalgae biomass in anaerobic cattle wastewater and the effects on lipids production

In the present study, the microalgae Spirulina platensis DRH 20 was cultivated in 4 photobioreactors with lighting intensity of 330 (±13.2) μmol m−2 s−1, 24 h d−1. No additives were added in PBR 1, while CO2 and phosphoric acid (pH control) were added in PBR 2, phosphoric acid was added in PBR 3, and CO2 was added in PBR 4. Dry biomass reached a maximum of 20.7 g L−1 and productivity reached a maximum of 4.3 g L−1 day−1 in PBR 2. The highest value of CO2 biofixation rate was also found in PBR 2, with 7.758 mg L−1 d−1. The highest μmax and the shortest doubling time were found in PBR 4. Regarding bioremediation, percentages between 76.9% and 79% of COD, 90.9 and 99.3% of NH4+, 85% and 92%. NO3−, were removed. The highest lipid production was recorded in PBR 4 (0.581 g L−1 d−1), which could generate 52.800 gallons of crude oil per year for each hectare cultivated and a biodiesel production yield of 121.7 g kg−1. From the analyses of fatty acids, it was possible to observe that producing biodiesel with the biomass generated in PBR 4 can promote better yields in lipid production and better quality.

Sequential adaptation to serum-free medium for Vero cells cultivation on ultraviolet/ozone (UVO) treated microcarrier

The adoption of serum-free media (SFM) in cell culture has gained prominence as an alternative to serum-supplemented media (SSM) owing to concerns regarding serum variability and quality assurance. Vero cells, widely utilized in viral vaccine production for their susceptibility to various viruses, were the focus of this study. Our primary objective was to adapt Vero cells from SSM to SFM and subsequently upscale the culture from T-flasks to spinner vessels using ultraviolet/ozone (UVO) treated polystyrene (PS) and polycaprolactone (PCL) microcarriers. Cell counting was conducted via haemocytometer and inverted phase microscopy. The results indicated that SFM-adapted Vero cells achieved higher maximum cell concentrations compared to those cultured in SSM, along with a shorter doubling time. However, during cultivation in spinner vessels, Vero cells exhibited lower maximum cell concentrations compared to T-flask cultures, attributed to factors such as cell inoculum size and microcarrier hydrophilicity. This study highlights the successful adaptation of Vero cells to SFM and the potential benefits in terms of cell density and growth kinetics. Furthermore, it sheds light on the challenges encountered during culture upscale, offering valuable insights for optimizing future large-scale production processes. The findings contribute to the advancement of serum-free cell culture systems and their applicability in the production of viral vaccines.

Pengaruh Perbedaan Jenis dan Konsentrasi Sumber Nitrogen (NaNO3 dan urea) terhadap Produksi Biomasa Spirulina Platensis

The goal of this study was to investigate the effect of the concentration and the type of nitrogen sources in the cultivation medium on the production of biomass and biochemical content (carbohydrates, proteins, fats) in the Spirulina platensis biomass. Types of nitrogen sources included NaNO3 and urea, where the concentrations were varied from 0.5 to 3.5 g/L. The results showed that the use of urea tended to produce more S. platensis biomass than the use of NaNO3. The best concentrations that produced the highest biomass were NaNO3 2.5 g/L (biomass 0.6745 g/L) and urea 0.5 g/L (biomass 0.7158 g/L). The use of urea also tended to produce a higher specific growth rate and a shorter doubling time than the use of NaNO3. The high concentration of nitrogen sources caused the pH of the medium to increase more rapidly due to the accumulation of ammonium in the medium. Using NaNO3 can produce biomass with a higher protein content (28.34-36.98%) than using urea (25.86-33.52%).

Methanol bioconversion in Methylotuvimicrobium buryatense 5GB1C through self-cycling fermentation

Methanol is an abundant and low-cost next-generation carbon source. While many species of methanotrophic bacteria can convert methanol into valuable bioproducts in bioreactors, Methylotuvimicrobium buryatense 5GB1C stands out as one of the most promising strains for industrialization. It has a short doubling time compared to most methanotrophs, remarkable resilience against contamination, and a suite of tools enabling genetic engineering. When approaching industrial applications, growing M. buryatense 5GB1C on methanol using common batch reactor operation has important limitations; for example methanol toxicity leads to mediocre biomass productivity. Advanced bioreactor operation strategies, such as fed-batch and self-cycling fermentation, have the potential to greatly improve the industrial prospects of methanotrophs growing on methanol. Herein, implementation of fed-batch operation led to a 26-fold increase in biomass density, while two different self-cycling fermentation (SCF) strategies led to 3-fold and 10-fold increases in volumetric biomass productivity. Interestingly, while synchronization is a typical trait of microbial populations undergoing SCF, M. buryatense 5GB1C cultures growing under this mode of operation led to stable, reproducible cycles but no significant synchronization.

Identification of Six Novel Prognostic Gene Signatures as Potential Biomarkers in Small Cell Lung Cancer.

As a subgroup of lung cancer, small cell lung cancer (SCLC) is characterized by a short tumor doubling time, high rates of early occurred distant cancer spread and poor outcomes. Our study aimed to identify novel molecular markers associated with SCLC prognosis. Microarray data from the Gene Expression Omnibus (GEO) database of SCLC tumors and paired normal tissues were obtained. In the dataset, Differentially expressed genes (DEGs) which were identified by comparing gene expression between normal lung and SCLC samples, were screened using the R language. The STRING database was used to map protein-protein interaction (PPI) networks, and these were visualized with the Cytoscape software. Go enrichment analysis and prediction were performed using the Metascape database and the results were visualized. Autophagy-related prognostic genes were identified by univariate COX regression analysis. Subsequently, stepwise model selection using the Akaike information criterion (AIC) and multivariate COX regression model was performed to construct DEGs signature. Survival receiver operating characteristic (ROC) analysis was used to assess the performance of survival prediction. At last, we evaluated the differences in drug sensitivity of the two groups of patients to common chemotherapeutic drugs and small-molecule targeted drugs. A total of 441 identified DE genes, including 412 downregulated and 29 upregulated genes were identified. GO enrichment analyses showed that DEGs were significantly enriched in the collagen-containing extracellular matrix and extracellular matrix organization. 16 genes were individually associated with OS in univariate analyses. The high expression of 6 genes (HIST1H4L, RP11-16O9.2, SNORA71A, SELV, FAM66A and BRWD1-AS1)) was associated with the poor prognosis of SCLC patients. To predict patients' outcomes, we developed an individual's risk score model based on the 6 genes. We found that SCLC patients with a low-risk score had significantly better survival than those with a high-risk score. What's more, association analysis between clinicopathological factors and gene signature showed the risk score was higher in patients with higher clinical stage or T stage. What's more, the patients in the high-risk score group had better treatment effects for etoposide and docetaxel. This suggests that our model can guide clinical treatment decisions. A novel six-gene signature was determined for prognostic prediction in SCLC. Our findings may provide new insights into the precise treatment and prognosis prediction of SCLC.

PD16-12 REAL-WORLD OUTCOMES AMONG MEN WITH NONMETASTATIC CASTRATION-SENSITIVE BIOCHEMICAL RECURRENCE AFTER DEFINITIVE THERAPY WITH SHORT PSA DOUBLING TIME RECEIVING EARLY SECONDARY TREATMENT

You have accessJournal of UrologyCME1 Apr 2023PD16-12 REAL-WORLD OUTCOMES AMONG MEN WITH NONMETASTATIC CASTRATION-SENSITIVE BIOCHEMICAL RECURRENCE AFTER DEFINITIVE THERAPY WITH SHORT PSA DOUBLING TIME RECEIVING EARLY SECONDARY TREATMENT Stephen J Freedland, Wei Gao, Angela Lax, Hongbo Yang, Krishnan Ramaswamy, David Russell, Agnes Hong, and Jasmine Ivanova Stephen J FreedlandStephen J Freedland More articles by this author , Wei GaoWei Gao More articles by this author , Angela LaxAngela Lax More articles by this author , Hongbo YangHongbo Yang More articles by this author , Krishnan RamaswamyKrishnan Ramaswamy More articles by this author , David RussellDavid Russell More articles by this author , Agnes HongAgnes Hong More articles by this author , and Jasmine IvanovaJasmine Ivanova More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003271.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: There is no consensus on treatment (tx) for men with nonmetastatic castration-sensitive prostate cancer (nmCSPC) with biochemical recurrence (BCR) after definitive tx (radical prostatectomy or radiotherapy). We assessed outcomes in men with nmCSPC and short prostate-specific antigen doubling time (PSADT) following BCR after definitive tx. METHODS: We reviewed the Veterans Health Administration (VHA) database between January 1, 2006 and June 22, 2020 for men with nmCSPC and BCR (index date) after definitive tx. Men were required to have PSADT ≤15 months after index date and were categorized into rapid (≤9 months) and less rapid (>9 but ≤15 months) PSADT. Outcomes included time to first antineoplastic tx after BCR and metastasis-free survival (MFS; time to metastasis or death). Cox proportional models compared the rapid and less rapid PSADT cohorts. Adjusted models controlled for baseline demographic and clinical characteristics. RESULTS: 753 men with nmCSPC were included in the analysis (rapid, n=485; less rapid, n=268). The rapid PSADT men were younger, more likely to be White, and had shorter median time from definitive tx to index (Table 1). Versus less rapid PSADT, rapid PSADT men had shorter median time to first antineoplastic tx (11.3 vs. 29.3 months), and MFS (94.1 vs. not reached) after BCR. CONCLUSIONS: Most rapid PSADT men underwent secondary tx within 1 year of BCR compared with less rapid PSADT men (53.2% vs 16.2%), whereas most less rapid PSADT men remained tx free for >2 years. Importantly, median MFS in men with rapid PSADT was 94.1 months, closely approximating the Pound data from Hopkins, wherein median time to metastases after BCR was 96 months (8 years). However, the Pound data included all BCR patients, not just those with rapid PSADT. Also, in the Pound data, no patients received early hormonal tx, whereas most VHA patients received secondary tx within 1 year of BCR. While circumstantial, the fact that rapid PSADT VHA patients heavily treated with secondary tx had similar outcomes to all BCR patients in the Pound series where none received aggressive secondary tx suggests the potential benefit of aggressive early tx of men with rapid PSADT after BCR. Source of Funding: Pfizer © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e493 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Stephen J Freedland More articles by this author Wei Gao More articles by this author Angela Lax More articles by this author Hongbo Yang More articles by this author Krishnan Ramaswamy More articles by this author David Russell More articles by this author Agnes Hong More articles by this author Jasmine Ivanova More articles by this author Expand All Advertisement PDF downloadLoading ...