NB Treatment Research Articles

Intravesical Botulin Toxin-A Injections for Neurogenic Bladder Dysfunction in Children: Summary Update on Last 10 Years of Research.

Neurogenic bladder dysfunction (NB) represents a challenge in pediatric urology. Intravesical botulin toxin-A (BTX-A) bladder injection is part of the armamentarium for the treatment of this condition, usually after failed first-line medical strategies and before the escalation to more invasive options such as neuromodulation or augmented cystoplasty in severe cases. However, there is still a lack of consensus about the appropriate treatment modality for the pediatric population. A review of the last 10 years' research was performed on the PubMed database by two authors. Articles doubly selected and meeting the inclusion criteria were collected and analyzed for their study type, demographics, neurological disease(s) at diagnosis, BTX-A treatment modality and duration, previous treatment, clinical and urodynamic parameters, adverse events, outcomes, and follow-ups. A total of 285 studies were initially selected, 16 of which matched the inclusion criteria. A cohort of 630 patients was treated with BTX-A at a median age of 9.7 years, 40% of which had a diagnosis of myelomeningocele. The results of the selected publications show the overall efficacy and safety of BTX-A injections in children and confirmed BTX-A as a valuable strategy for NB treatment in pediatric population. Nevertheless, up to now, the literature on this topic offers scarce uniformity among the published series and poor protocol standardization.

Anti-Inflammatory Efficacy of Nanobody Specific to β-Glucan on a Fungal Cell Wall in a Murine Model of Fungal Keratitis.

Purpose: to explore the anti-inflammatory effects of a nanobody (Nb) specific to β-glucan on fungal keratitis (FK). Methods: in order to verify the therapeutic and anti-inflammatory efficacy of Nb in FK, the severity of inflammation was assessed with inflammatory scores, hematoxylin-eosin (HE) staining, and myeloperoxidase (MPO) assays. In corneas of mice of FK model and human corneal epithelial cells stimulated by fungal hyphae, real-time reverse transcriptase polymerase chain reaction, Western blot, and enzyme-linked immunosorbent assay were used to detect the expression levels of inflammatory cytokines and pattern recognition receptors (PRRs). In vivo, macrophages and neutrophils infiltration in the cornea stroma was detected by immunofluorescence (IFS) staining. Results: In murine models infected with Aspergillus fumigatus (A. fumigatus), Nb treatment could reduce the inflammatory scores. HE staining and MPO results showed Nb significantly alleviated corneal edema and reduced inflammatory cell infiltration 3 days post-infection. In addition, the expression levels of LOX-1 and Dectin-1 were significantly decreased in the Nb group in vivo. The expression of chemokines CCL2 and CXCL2 also decreased in the Nb group. Compared with the PBS group, the number of macrophages and neutrophils in the Nb group was significantly decreased, which was shown in IFS results. Moreover, Nb attenuated the expression of Dectin-1, LOX-1, and inflammatory mediators, including IL-6 and IL-8 in vitro. Conclusion: our study showed that Nb could alleviate FK by downregulating the expression of PRRs and inflammatory factors as well as reducing the infiltration of macrophages and neutrophils.

One–third substitution of nitrogen with cow manure or biochar greatly reduced N2O emission and carbon footprint in saline–alkali soils

The rapid expansion of farmland and long−term excessive nitrogen (N) application have caused huge environmental risks in fragile ecosystems facing global warming. Partially substituting N fertilizer with organic fertilizers offers an alternative field management strategy to alleviate the pressure of the ecological environment. To this end, the influence of one−third substitution of N fertilizer with cow manure or biochar field experiment was conducted under maize in Tarim River Basin since 2019. Five treatments with three replications were applied: CK (Fallow); no fertilization (0 N); conventional N fertilizer (N; N: 300 kg N ha−1, organic fertilizer: 0 kg N ha−1); one−third substitution of N with biochar (NB; N: 200 kg N ha−1, Biochar: 100 kg N ha−1) and one−third substitution of N with cow manure (NM; N: 200 kg N ha−1, Cow manure: 100 kg N ha−1) under maize season in saline−alkali soils. The greenhouse gas (GHG) emissions, net ecosystem carbon budget (NECB), soil organic carbon (SOC), maize yield, carbon footprint (CF), and yield carbon footprint (CFy) were analyzed from 2021 to 2022. The results showed that NB treatment decreased the average cumulative CO2 emissions by 21 %, while NM treatment showed no difference compared to N treatment. NB and NM treatments reduced the average cumulative N2O emissions (−61 %, −49 %), CF (−68 %, −10 %), CFy (−66 %, −19 %) and increased maize yield (+3 %, +2 %), SOC storage (+43 %, +6 %), NECB (+80 %, +24 %), and agronomic N use efficiency (ANUE) (+5 %, +3 %), compared to N treatment. NB treatment had the lowest emission factors (EF) (0.19 %) and the highest sustainability index (1.58) compared to NM treatment (0.26 %, 0.61) and N treatment (0.53 %, 0.84). To sum up, substituting one−third of N fertilizer with biochar or manure in saline−alkali soils was proved to be a multi−benefit strategy to increase yields and reduce GHG emissions and CF.

Influence of NUCB/Nesfatin-1 Polymorphism on Treatment Response to Naltrexone/Bupropion SR in Binge Eating Disorder and Obesity.

Background and Objectives: The NUCB2 gene and its polymorphisms were identified as novel players in the regulation of food intake, potentially leading to obesity (OBE) and altered eating behaviors. Naltrexone/bupropion SR (NB) showed good efficacy and tolerability for treating OBE and altered eating behaviors associated with binge eating disorder (BED). This prospective study investigates the influence of NUCB2 gene polymorphism on NB treatment response in OBE and BED. Materials and Methods: Body mass index (BMI), eating (EDE-Q, BES, NEQ, GQ, Y-FAS 2.0) and general psychopathology (BDI, STAI-S) were evaluated at baseline (t0) and after 16 weeks (t1) of NB treatment in patients with OBE and BED (Group 1; N = 22) vs. patients with OBE without BED (Group 2; N = 20). Differences were evaluated according to the rs757081 NUCB2 gene polymorphism. Results: NUCB2 polymorphism was equally distributed between groups. Although weight at t0 was higher in Group 1, weight loss was similar at t1 in both groups. BMI was not influenced by NUCB2 polymorphism. In Group 1, the CG-genotype reported significant improvement in eating psychopathology while the GG-genotype reported improvement only for FA. No differences were observed in Group 2. Conclusions: Patients diagnosed with BED and treated with NB exhibited a more favorable treatment response within the CG-genotype of the NUCB2 polymorphism.

N-Butanol fraction of moringa seed attenuates arsenic intoxication by regulating the uterine inflammatory and apoptotic pathways.

The adverse effects of arsenic-chelating drugs make it essential to replace invasive chelating therapy with non-invasive oral therapy for arsenic poisoning. The goal of the current investigation was to determine whether the uterine damage caused by arsenization could be repaired by the n-butanol fraction of Moringa oleifera seed (NB). The rats were orally administered with arsenic (10mg/kg BW) for the initial 8days, followed by NB (50mg/kg) for the next 8days without arsenic. The probable existence of different components in NB was evaluated by HPLC-MS. Pro and anti-inflammatory indicators were assessed by RT-PCR and western blot. ESR-α was detected via immunostaining. Arsenic-exposed rats had significantly increased lipid peroxidation and decreased antioxidant enzyme activity, which were markedly reduced after NB treatment. Weaker ESR-α expression and distorted uterine histomorphology following arsenication were retrieved significantly by NB. Meaningful restoration by NB was also achieved for altered mRNA and protein expression of various inflammatory and apoptotic indicators. Molecular interaction predicted that glucomoringin and methyl glucosinolate of moringa interact with the catalytic site of caspase-3 in a way that limits its activity. However, NB was successful in restoring the arsenic-mediated uterine hypofunction. The glucomoringin and methyl glucosinolate present in n-butanol fraction may play a critical role in limiting apoptotic event in the arsenicated uterus.

The preparation and therapeutic effects of β-glucan-specific nanobodies and nanobody-natamycin conjugates in fungal keratitis.

Fungal keratitis (FK) is a severe infectious corneal disease. Since traditional eye drops exhibit poor dissolution and high corneal toxicity, the efficacy of current treatments for FK remains limited. It is needed to develop new approaches to control the cornea damage from FK. In this study, a nanobody (Nb) specific to β-glucan in the fungal cell wall was prepared. The conjugate of the Nb with natamycin (NAT), a traditional antifungal drug, was synthesized. Firstly, we found the Nb specific to β-glucan inhibited fungal growth by disrupting cell wall and biofilm formation.. In addition, the content of β-glucan in the fungal cell wall decreased after Nb treatment. The Nb also reduced the adhesion ability of fungal conidia to human corneal epithelial cells (HCECs). Further, we examined the difference between NAT and Nb-NAT in antifungal growth. Nb-NAT showed better antifungal effects than NAT which was caused by the interaction between Nb and β-glucan. Moreover, Nb concentration below 0.5 mg/mL did not affect the viability of HCECs. Nb-NAT had less cytotoxicity and ocular surface irritation than NAT. Nb specific to β-glucan attenuated Aspergillus fumigatus (A. fumigatus) virulence and relieved inflammatory responses in FK. Nb-NAT treatment of the cornea improved therapeutic effects compared with NAT. It decreased clinical scores and expression level of inflammatory factors. To our knowledge, this study is the first to report a Nb specific to β-glucan and Nb-NAT for the treatment of FK. These unique functions of the Nb specific to β-glucan and Nb-NAT would render it as an alternative molecule to control fungal infections including FK. STATEMENT OF SIGNIFICANCE: Fungal keratitis is a corneal disease with a high rate of blindness. Due to the poor dissolution and high corneal toxicity exhibited by traditional eye drops, the efficacy of current therapeutic treatments for fungal keratitis (FK) remains limited. To enhance the therapeutic effect of natamycin in treating fungal keratitis, this study developed an innovative approach by preparing a β-glucan-specific nanobody and loading it with the antifungal drug natamycin. The β-glucan-specific nanobody has the ability to control both fungal pathogen invasion and inflammation, which can cause damage to the cornea in FK. The conjugation with the β-glucan-specific nanobody significantly increased the antifungal capacity of natamycin and reduced its toxicity. The further application of natamycin conjugated with the β-glucan-specific nanobody could be expanded to other diseases caused by fungal pathogen infections.

Blocking P2X7 by intracerebroventricular injection of P2X7-specific nanobodies reduces stroke lesions

BackgroundPrevious studies have demonstrated that purinergic receptors could be therapeutic targets to modulate the inflammatory response in multiple models of brain diseases. However, tools for the selective and efficient targeting of these receptors are lacking. The development of new P2X7-specific nanobodies (nbs) has enabled us to effectively block the P2X7 channel.MethodsTemporary middle cerebral artery occlusion (tMCAO) in wild-type (wt) and P2X7 transgenic (tg) mice was used to model ischemic stroke. Adenosine triphosphate (ATP) release was assessed in transgenic ATP sensor mice. Stroke size was measured after P2X7-specific nbs were injected intravenously (iv) and intracerebroventricularly (icv) directly before tMCAO surgery. In vitro cultured microglia were used to investigate calcium influx, pore formation via 4,6-diamidino-2-phenylindole (DAPI) uptake, caspase 1 activation and interleukin (IL)-1β release after incubation with the P2X7-specific nbs.ResultsTransgenic ATP sensor mice showed an increase in ATP release in the ischemic hemisphere compared to the contralateral hemisphere or the sham-treated mice up to 24 h after stroke. P2X7-overexpressing mice had a significantly greater stroke size 24 h after tMCAO surgery. In vitro experiments with primary microglial cells demonstrated that P2X7-specific nbs could inhibit ATP-triggered calcium influx and the formation of membrane pores, as measured by Fluo4 fluorescence or DAPI uptake. In microglia, we found lower caspase 1 activity and subsequently lower IL-1β release after P2X7-specific nb treatment. The intravenous injection of P2X7-specific nbs compared to isotype controls before tMCAO surgery did not result in a smaller stroke size. As demonstrated by fluorescence-activated cell sorting (FACS), after stroke, iv injected nbs bound to brain-infiltrated macrophages but not to brain resident microglia, indicating insufficient crossing of the blood–brain barrier of the nbs. Therefore, we directly icv injected the P2X7-specific nbs or the isotype nbs. After icv injection of 30 µg of P2X7 specific nbs, P2X7 specific nbs bound sufficiently to microglia and reduced stroke size.ConclusionMechanistically, we can show that there is a substantial increase of ATP locally after stroke and that blockage of the ATP receptor P2X7 by icv injected P2X7-specific nbs can reduce ischemic tissue damage.

Antibiotic therapy of neuroborreliosis: A survey among infectious disease specialists and neurologists in Norway, Sweden, and Denmark

IntroductionNeuroborreliosis (NB) is a prevalent tick-borne neuroinfection in Europe. To delineate current practice in antimicrobial management of adults with NB and to prioritize future trials needed to optimize treatment recommendations, a questionnaire-based survey was performed. MethodsA self-administered Internet-based survey of NB treatment practices among specialists in infectious diseases and neurology based in Norway, Sweden, and Denmark was carried out between October 2021 and February 2022. The participants were also asked to prioritize four pre-defined research questions for randomized controlled trials (RCTs) on therapy for NB. ResultsIn total, 290 physicians (45% female) from Norway (30%), Sweden (40%), and Denmark (30%) participated in the survey. Of the responders, 230 (79%) were infectious disease specialists and 56 (19%) were neurologists.The preferred antibiotic treatment for patients with early NB was oral doxycycline (n = 225, 78%). Intravenous (IV) penicillin, ceftriaxone, or cefotaxime for the full treatment course was favored by 12%. A preferred treatment duration of 10–14 days for patients with NB was reported by 245 respondents (85%), most common among participants from Sweden (97%).A total of 170 (59%) responders reported having local hospital guidelines on the treatment of NB, most often with recommendation of oral doxycycline (92%) for 10–14 days (90%) as first line treatment.The prioritization score for future RCTs was highest for adjunctive prednisone therapy in NB patients with facial palsy (median 5; IQR 4–6) and for placebo versus repeated antibiotics in patients with persistent symptoms after completed antibiotic therapy for NB (median 5, IQR 3–6). ConclusionIn Sweden, all respondents preferred treating NB with oral doxycycline for 10–14 days, whereas 5% in Norway and 19% in Denmark still treat NB with IV antibiotics for the entire treatment course. RCTs to define the role of adjunctive prednisolone in NB patients with facial palsy and repeated antibiotics in patients with persistent symptoms are prioritized for future research.

Contrasting effects of different field-aged biochars on potential methane oxidation between acidic and saline paddy soils

There is recognition that biochar addition is an appropriate measure to mitigate methane (CH4) emissions by promoting potential methane oxidation (PMO) in the field. However, the mechanism for different field-aged biochars and effective duration after field application are not well documented. Based on a long-term field experiment, biochar was field aged and separated from two contrasting acidic (Ba) and saline (Bs) paddy fields. Then, the effects of different aged biochars on PMO in acidic and saline paddy soils were explored by incubation experiment. There were five treatments for each soil group: soil without biochar (CK), biochar-enriched paddy soil (2 or 6 years) (NB), fresh biochar amendment (Bf), aged biochar separated from acidic paddy soil amendment (Ba), and aged biochar separated from saline paddy soil amendment (Bs). Results showed that saline paddy soils had a significantly higher PMO than acidic paddy soils under treatment without biochar, and that PMO in acidic paddy soil was enhanced by various biochar amendments, whereas those biochar amendments had no significant effects on PMO in saline paddy soil. PMO was positively correlated with pmoA abundance, N consumption rate and pH of soil-biochar mixture. Aged biochar separated from different fields had conflicting influences on soil pH, N consumption rate and PMO. Ba lost its initial effect on changing PMO as compared to Bf treatment when added back into acidic paddy soil. To the contrary, the acidic paddy soil NB treatment containing biochar added six years before possessed the highest value of PMO among all ten treatments. This study suggested that acidic paddy soil with biochar amendment could mitigate CH4 emissions by promoting PMO for a prolonged period, though aged biochar separated from the same field had a limited impact on reducing CH4 emissions.

Abstract 5719: Defining TCF4’s role as a novel key regulator in mediating neuroblastoma cell identity

Abstract Neuroblastomas (NB) are embryonal childhood tumors that derive from the multipotent neural crest cells (NCCs) of the peripheral nervous system. NB accounts for more than 15% of all childhood cancer-related deaths. Despite the most intensive multimodal therapy, more than 50% of patients with high-risk NB relapse with often fatal, resistant disease. Finding novel treatments, especially for relapse disease, is desperately needed for high-risk NB. In cancers, distinct transcription factors TFs networks forming core regulatory circuitries (CRCs) control gene expression programs that drive cell identity. Recent studies reported the presence of two types of identity states in NB tumors: one establishing a more proliferative adrenergic (ADRN) cell state and a second establishing a more invasive, therapy-resistant mesenchymal (MES) cell state. While most studies are investigating the CRC members for targeting MYCN amplified NB (ADRN) or the MES/NCCs cell states, most primary tumors are heterogeneous, comprised of cells with both identities. Thus, our work is focused on identifying universal factors shared across different lineage states to determine whether targeting both identities could be a valuable strategy for NB treatment. NB was identified from a screen of 673 cancer cell lines as highly sensitive to the BET-inhibitor JQ1. In our work, we identified the class I basic helix-loop-helix transcription factor (bHLH) TCF4 as a critical target of JQ1 mediated cell death in NB. TCF4 functions as a transcriptional hub that heterodimerizes with class II (bHLH) TFs including the proneural gene ASCL1 (ADRN), and TWIST1 a master regulator of the MES state. Heterodimers formed between TCF4 and ASCL1, and TWIST1, have been demonstrated to provide lineage-specific differentiation from embryonic stem cells. Here, we hypothesize that TCF4 is a cell dependency gene in NB that is crucial for determining NB identity. Using RNA seq analysis of TCF4 dox-inducible shRNA stable cell lines, we found that the gene expression changes in TCF4-depleted cells indicate a role for TCF4 in differentiation, cell signaling, and neurodevelopment. Interestingly, gene set enrichment analysis (GSEA) indicates that both identity states are suppressed after silencing of TCF4. Our data also shows that inducible decrease of TCF4 protein resulted in a significant inhibition of NB cell growth, and impaired tumor growth in vivo. Furthermore, we identified TCF4 targets in NB by combined analysis of TCF4 ChIP-seq data and gene expression changes following TCF4 knockdown. ENRICHR analysis suggests that differentially expressed genes in TCF4-knockdown cells that also have a TCF4 ChIP-seq peak are targets of the TFs TWIST1 (MES) and HAND1 (ADRN) that interact directly with TCF4. In conclusion, our preliminary data supports the hypothesis that TCF4 is a master transcriptional regulator of NB oncogenic program that is crucial for maintaining NB identity states. Citation Format: Nour Aljouda, Dewan Shrestha, Megan Walker, Satyanarayana Alleboina, Evan Glazer, Yong Cheng, Kevin Freeman. Defining TCF4’s role as a novel key regulator in mediating neuroblastoma cell identity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5719.

Effects of biochar combined with nitrification/urease inhibitors on soil active nitrogen emissions from subtropical paddy soils

We examined the effects of biochar and urease inhibitors/nitrification inhibitors on nitrification process, ammonia and N2O emission in subtropical soil, and determined the best combination of biochar with nitrification and urease inhibitors. This work could provide a theoretical basis for the mitigation of the negative environmental risk caused by reactive nitrogen gas in the application of nitrogen fertilizer. A indoor aerobic culture test was conducted with seven treatments [urea+biochar (NB), urea+nitrification inhibitor (N+NI), urea+urease inhibitor (N+UI), urea+nitrification inhibitor+urease inhibitor (N+NIUI), urea+nitrification inhibitor+biochar (NB+NI), urea+urease inhibitor+biochar (NB+UI), urea+nitrification inhibitor+urease inhibitor+biochar (NB+NIUI)] and urea (N) as the control. The dynamics of soil inorganic nitrogen content, N2O emission and the volatility of ammonia volatilization were observed under combined application of biochar with urease inhibitor (NBPT)/nitrification inhibitor (DMPP). The results showed that:1)Compared to the control (5.11 mg N·kg-1·d-1) during the incubation period, NB treatment significantly increased therate constant of nitrification by 33.9%, and N+NI treatment significantly reduced the nitrification rate constant by 22.9%. NB treatment significantly increased the abundance of ammonia oxidizing bacteria (AOB) by 56.0%. 2) Compared with N treatment, N+NI and NB+NI treatments signi-ficantly enhanced the cumulative emission of NH3 by 49%. The N+UI treatment reduced the cumulative loss of NH3. The inhibition effect of NB+UI treatment was more significant. 3) The emission rate of N2O was highest in the first 10 days after fertilization. The N2O emission under NB treatment was the earliest, and that of N treatment was the highest (5.87 μg·kg-1·h-1). The combined application of DMPP and NBPT performed the best in reducing soil N2O emission. We estimated global warming potential (GWP) of the direct N2O and indirect N2O (NH3) emissions. Compared with N treatments, N+NI and NB+NI treatments increased the GWP by 34.8% and 40.9%, respectively. While the NB and NB+UI treatments significantly reduced the GWP by 45.9% and 60.5%, the combination of biochar and urease inhibitor had the best effect on reduction of GWP of soil active nitrogen emissions.

Integrated analysis of the functions and prognostic values of RNA-binding proteins in neuroblastoma.

BackgroundNeuroblastoma (NB) is the most common solid tumor in children. NB treatment has made significant progress; however, given the high degree of heterogeneity, basic research findings and their clinical application to NB still face challenges. Herein, we identify novel prognostic models for NB.MethodsWe obtained RNA expression data of NB and normal nervous tissue from TARGET and GTEx databases and determined the differential expression patterns of RNA binding protein (RBP) genes between normal and cancerous tissues. Lasso regression and Cox regression analyses identified the five most important differentially expressed genes and were used to construct a new prognostic model. The function and prognostic value of these RBPs were systematically studied and the predictive accuracy verified in an independent dataset.ResultsIn total, 348 differentially expressed RBPs were identified. Of these, 166 were up-regulated and 182 down-regulated RBPs. Two hubs RBPs (CPEB3 and CTU1) were identified as prognostic-related genes and were chosen to build the prognostic risk score models. Multivariate Cox analysis was performed on genes from univariate Cox regression and Lasso regression analysis using proportional hazards regression model. A five gene prognostic model: Risk score = (-0.60901*expCPEB3)+(0.851637*expCTU1) was built. Based on this model, the overall survival of patients in the high-risk subgroup was lower (P = 2.152e-04). The area under the curve (AUC) of the receiver-operator characteristic curve of the prognostic model was 0.720 in the TARGET cohort. There were significant differences in the survival rate of patients in the high and low-risk subgroups in the validation data set GSE85047 (P = 0.1237e-08), with the AUC 0.730. The risk model was also regarded as an independent predictor of prognosis (HR = 1.535, 95% CI = 1.368–1.722, P = 2.69E-13).ConclusionsThis study identified a potential risk model for prognosis in NB using Cox regression analysis. RNA binding proteins (CPEB3 and CTU1) can be used as molecular markers of NB.

Optimizing Tomato Growth and Productivity Using Nitrogen and Irrigation Application Timing

Soil nutrients and water management practices according to the concept of 4R nutrient stewardship (right rate, right timing, right placement, and right source) can have significant benefits on crop productivity and reduce the negative effects of agricultural practices on the environment. Therefore, this present study evaluated the effects of nitrogen (N) application timing under different irrigation regimes on open-field, fresh-market tomato production in Florida. In this study, 2 N application timings applied at 25% pre-plant with 75% fertigation (BM), and 0 pre-plant with 100% fertigation (NB), were evaluated. The two N application methods were evaluates using three irrigation regimes: full irrigation (FI, 100% ETc), deficit irrigation (DI, 66% ETc), and regulated deficit irrigation (RDI, 66% ETc during the first 4 weeks after transplanting and 100% ETc afterward). The results showed that BM treatment significantly improved early-season tomato growth compared to NB treatment. The results also indicated that under RDI and DI irrigation conditions, tomato root length was lowest (average value of 13%) within the first 15 cm compared to 40% within 15–30 cm and 47% at 30–40 cm soil depths. Similar to plant growth, BM treatment significantly increased tomato yield (average valued 56.00 Mg ha−1) compared to the NB (average value 40.23 Mg ha−1). The application of DI throughout the growing season reduced tomato yield; however, there were no differences in yield under the RDI and FI irrigation regimes. Therefore, based on the results from this study, it can be concluded that, under Florida growing conditions, pre-plant N application is essential for tomato growth and productivity. Additionally, irrigation application using the RDI method could be successfully adopted in Florida tomato production for improved water savings without any negative effects on tomato growth and productivity.

Abstract 1544: Generation of ALK CAR-T cells for neuroblastoma therapy

Abstract Neuroblastoma (NB) is the most deadly cancer in children with dismal survival in high-risk patients. The majority of NB express the full length anaplastic lymphoma kinase (ALK) receptor, that typically acts as driver oncogene together with MYCN. In contrast to ALK-driven lung cancer or lymphoma, targeted therapies with ALK tyrosine kinase inhibitors (TKIs), despite encouraging, induce only partial responses in NB. Therefore, additional tools to improve NB treatment are strongly needed. To specifically target NB cells, we developed a series of ALK chimeric antigen receptor (CAR) T constructs from antibodies that recognize both human and mouse ALK. Murine ALK CAR-T cells are able to control the growth of ALK+ leukemia and NB in syngeneic tumor models without detectable toxicity. From the leading candidate, we generated fully humanized ALK CAR-T cells that showed potent in vitro killing activity against a large panel of human NB lines, with activity comparable to GD2 CAR-T cells that are currently in clinical trials for NB patients. Remarkably, ALK CAR-T treatment synergized in vivo with the ALK inhibitor lorlatinib. Mechanistically, lorlatinib not only reduced tumor growth, but enhanced ALK expression on the surface of tumor cells, thereby facilitating ALK CAR-T targeting. Combination of ALK CAR-T cells with lorlatinib resulted in enhanced killing of NB cells and cure or markedly increased survival in mouse models of human metastatic NB even with low ALK expression. These findings support the clinical development of ALK CAR-T cells for NB therapy. Citation Format: Elisa Bergaggio, Wei-Tien Tai, Andrea Aroldi, Ineês Mota, Diego Alvarado, Elisa Landoni, Jasna Metovic, Manuel Nüesch, Rafael Blasco-Patiño, Mauro Papotti, Gianpietro Dotti, Roberto Chiarle. Generation of ALK CAR-T cells for neuroblastoma therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1544.

Prognostic Signature of Immune Genes and Immune-Related LncRNAs in Neuroblastoma: A Study Based on GEO and TARGET Datasets.

BackgroundThe prognostic value of immune-related genes and lncRNAs in neuroblastoma has not been elucidated, especially in subgroups with different outcomes. This study aimed to explore immune-related prognostic signatures.Materials and MethodsImmune-related prognostic genes and lncRNAs were identified by univariate Cox regression analysis in the training set. The top 20 C-index genes and 17 immune-related lncRNAs were included in prognostic model construction, and random forest and the Least Absolute Shrinkage and Selection Operator (LASSO) regression algorithms were employed to select features. The risk score model was constructed and assessed using the Kaplan-Meier plot and the receiver operating characteristic curve. Functional enrichment analysis of the immune-related lncRNAs was conducted using the STRING database.ResultsIn GSE49710, five immune genes (CDK4, PIK3R1, THRA, MAP2K2, and ULBP2) were included in the risk score five genes (RS5_G) signature, and eleven immune-related lncRNAs (LINC00260, FAM13A1OS, AGPAT4-IT1, DUBR, MIAT, TSC22D1-AS1, DANCR, MIR137HG, ERC2-IT1, LINC01184, LINC00667) were brought into risk score LncRNAs (RS_Lnc) signature. Patients were divided into high/low-risk score groups by the median. Overall survival and event/progression-free survival time were shortened in patients with high scores, both in training and validation cohorts. The same results were found in subgroups. In grouping ability assessment, the area under the curves (AUCs) in distinguishing different groups ranged from 0.737 to 0.94, better in discriminating MYCN status and high risk in training cohort (higher than 0.9). Multivariate Cox analysis demonstrated that RS5_G and RS_Lnc were the independent risk factors for overall and event/progression-free survival (all p-values <0.001). Correlation analysis showed that RS5_G and RS_Lnc were negatively associated with aDC, CD8+ T cells, but positively correlated with Th2 cells. Functional enrichment analyzes demonstrated that immune-related lncRNAs are mainly enriched in cancer-related pathways and immune-related pathways.ConclusionWe identified the immune-related prognostic signature RS5_G and RS_Lnc. The predicting and grouping ability is close to being even better than those reported in other studies, especially in subgroups. This study provided prognostic signatures that may help clinicians to choose optimal treatment strategies and showed a new insight for NB treatment. These results need further biological experiments and clinical validation.

Comparison of microstructure and heat treatment distortion of gear steels with and without Nb addition

In order to research the effect of microalloying Nb, the microstructure and heat treatment distortion of case carburizing steels with and without Nb addition are compared. Results show that a uniform and fine prior austenite grain size was obtained for the steel with the addition of 0.03 wt.% Nb even after being carburized at 980 °C for 37 h, resulting in a very deep hardened layer of about 4 mm. Nb is an effective microalloying element to hinder austenite grain growth of gears during high-temperature carburizing. Theoretical calculation and experimental observation of NbC precipitates indicate that fine NbC precipitates have not evidently dissolved at 980 °C, and thus they can act as grain refiners due to pinning effect. Heat treatment distortion of Nb-added steel is much lower than that of the steel without Nb addition. It may be contributed to its fine and uniform grain size, which presumably influences stress during martensitic transformation.

Yield components of soybean cover crop regard to seed pre-treatment with bacteria and mycorrhiza

The aim of this study was to describe the importance of bacterization of soybean seeds and the use of preparations of mycorrhizal fungi in the sowing of soybeans on the family farm "Alduk" in 2020. Post- sowing of soybeans was done on 26th June 2020 as cover crop. Two very early varieties (00 maturity group): Korana (Agricultural Institute Osijek) and Merkur (NS seeds, Serbia) were used. Before sowing, the seeds were bacterized (Nitrobacterin – Faculty of Agrobiotechnical Sciences Osijek) or the addition of preparations of mycorrhizal fungi (VAM + Azotobacter chroococcum – Faculty of Agrobiotechnical Sciences Osijek). At harvest, yield components of soybeans in 2020 were determined. To determine the yield components from each treatment, 20 plants were selected and analysed separately. A total of 120 individual plants were analysed, and the following were determined: plant height (cm) and height to the first pod, number of fertile levels per plant from the central stem and per plant, number of pods per plant and seed mass of one plant (g), 1000 grain mass and at final, seed yield (t/ha). The height of the plants up to the first fertile pod was on average 7 cm, and varied from 5 cm (Mercury variety with VAM + AC treatment), to 9 cm (Korana on NB treatment). The number of fertile levels per plant averaged 11 on the main, central stem, while the total number of fertile levels per plant was 16. The number of pods per plant in this study averaged 42, with the seed weight of one plant being 10.48 g per plant. The highest mass of seeds per plant (g) had the Korana variety on the control treatment (14.95 g per plant). The Korana variety also had the lowest seed mass per plant (7.07 g per plant) with the application of Nitrobacterin. According to the results, Korana variety had the highest yield on the control treatment (1.19 t/ha), followed by the treatment with VAM + Azotobacter chroococcum (1.04 t/ha) and the lowest with the application of Nitrobacterin (0.84 t/ha). Merkur variety had the lowest soybean yield on the control treatment (0.69 t/ha), while with Nitrobacterin and VAM + Azotobacter chroococcum soybean yield increased by about 19% with Nitrobacterin and about 27% with VAM + Azotobacter chroococcum.

Analysis of treatment cost for neuroblastoma to the family: a single-center cross-sectional study in China

BackgroundNeuroblastoma (NB) is notorious in childhood cancer because of its high incidence and poor prognosis. The Children’s Oncology Group reported that the 3-year OS in the high-risk (HR) group is 50%, and the HR-NB with bone marrow metastasis in our center is 43.1%. Thousands of families in China suffer from the cost of NB, but the true costs of therapy are unknown; to date, no study has ever performed a detailed therapy costs analysis for NB. The objective of this study was to assess the economic burden of NB treatment in children to the family, to ultimately reduce related expenses for patients and promote the establishment of NB management policy.Materials and methodsData in this cross-sectional study were collected via questionnaires completed by parents at the outpatient clinic and was verified via a computer system. Therapy costs of children with NB of differing risks were analyzed through descriptive statistics (1 CNY ≈ 0.1412 USD).ResultsMedian direct medical costs of low risk (LR), middle risk (MR), and HR NB during treatment were 180.0 (120.0, 300.0), 200.0 (166.0, 300.0), and 650.0 (415.5, 850.0) thousand Chinese yuan (CNY), respectively. Direct non-medical costs including transportation, food, and accommodation were 60.0 (37.0, 100.0), 80.0 (60.0, 120.0), and 100.0 (80.0, 157.5) thousand CNY in the LR, MR, and HR groups, respectively. Additionally, parents accrued work absences to attend treatment, and lost a total of 100.0 (50.0, 150.0) thousand CNY in indirect costs.LimitationsFamilies whose children had relapsed or died were excluded from this analysis and therefore limited the conclusions drawn. Parents were asked to recall costs since initial diagnosis (1–6 years in the past), but this extended time period may have introduced recall bias.ConclusionsDirect non-medical and indirect costs play an important role in the total treatment costs of NB. Children with NB treated in local hospitals and followed up in hospital specialized in childhood oncology may save many unnecessary expenses. China's healthcare system should establish mechanisms and provide financial support for children with NB.

Hard and wear-resistant niobium, molybdenum carbide layered coatings on WC-Co tools produced by ion bombardment and cathodic vacuum arc deposition

Phase and element composition, microhardness, adhesive strength, wear resistance and microstructure of the coatings and underlying layers formed on WC-Co hard alloy using Nb and Mo vacuum-arc sources were studied. It is shown that the influence of ion beams with energy of 1 keV created by Nb and Mo vacuum-arc sources for at least 1–2 min allows the formation of carbide coatings based on embedded metals and alloy elements with (Nb,W)C0.7, Mo2C and Co6W6C2 phases. The synthesis of carbide layers occurred by ion exposure by Nb or Mo at a surface temperature of the hard alloy of at least 1300 °C, controlled by an optical thermograph. Reducing of Nb and Mo ion exposure time down to 30 s does not allow heating of samples surface to temperatures above 700 °C, as a result, the formation of carbide layers on the alloy surface does not occur. The plasma deposition of NbC or MoC coatings on sublayers created by preliminary Nb or Mo ion treatment accompanied by heating of the sample surface up to 1300°С forms carbide layered system with hardness of 60 GPa, adhesive strength of 120–140 N and 2–3-fold lower volume wear compared to NbC and MoC plasma coatings deposited on a hard alloy without preliminary treatment.

Long-Term Fire Regime Modifies Carbon and Nutrient Dynamics in Decomposing Eucalyptus pilularis Leaf Litter

The changes in fire regimes expected under climate change are likely to disrupt the biogeochemical cycling of carbon (C) and nutrients in forest ecosystems. Plant litter decomposition is a critical step in the terrestrial biogeochemical cycle, and is an important determinant of fire fuel load and forest C balance. We conducted a 277-day leaf litter decomposition experiment in an Australian eucalypt forest to test whether three contrasting, long-term fire regimes (no burning [NB], 4-yearly burning, and 2-yearly burning) were associated with different C and nutrient dynamics during litter decomposition. Fire regime had strong effects on many litter properties, including overall rates of decomposition and C loss, which were greatest in the NB treatment, suggesting that fire regime can modify the rate at which C is returned from litter to soil or the atmosphere. This has potentially important implications for soil C storage and atmospheric CO2 concentrations under a changing climate. High-frequency fire was associated with litter nutrient depletion and high microbial nutrient demand, but did not affect nutrient loss rates from decomposing litter, suggesting conservative use and retention of nutrients by the litter microbial biomass. These effects differed qualitatively between 2- and 4-yearly burning regimes, and they show how decadal-scale increases in fire frequency might contribute to soil nutrient depletion by disrupting decomposition. Many effects of fire regime on litter properties throughout decomposition were sensitive to litter bag re-collection date, suggesting that seasonal factors moderate the effects of fire regime, and that the role of fire regime-altered litter chemistry in shaping decomposition may be secondary to that of fire regime-altered environmental variables. Together, our findings highlight the potential consequences of long-term increases in prescribed fire frequency for litter decomposition and the storage and cycling of C and nutrients in eucalypt forests, and reveal the specific importance of average burn frequency in this context.