Targeting Alpha-Ketoglutarate Disruption Overcomes Immunoevasion and Improves PD-1 Blockade Immunotherapy in Renal Cell Carcinoma.

The Warburg effect-related metabolic dysfunction of the tricarboxylic acid (TCA) cycle has emerged as a hallmark of various solid tumors, particularly renal cell carcinoma (RCC). RCC is characterized by high immune infiltration and thus recommended for immunotherapeutic interventions at an advanced stage in clinical guidelines. Nevertheless, limited benefits of immunotherapy have prompted investigations into underlying mechanisms, leading to the proposal of metabolic dysregulation-induced immunoevasion as a crucial contributor. In this study, a significant decrease is found in the abundance of alpha-ketoglutarate (αKG), a crucial intermediate metabolite in the TCA cycle, which is correlated with higher grades and a worse prognosis in clinical RCC samples. Elevated levels of αKG promote major histocompatibility complex-I (MHC-I) antigen processing and presentation, as well as the expression of ß2-microglobulin (B2M). While αKG modulates broad-spectrum demethylation activities of histone, the transcriptional upregulation of B2M is dependent on the demethylation of H3K4me1 in its promoter region. Furthermore, the combination of αKG supplementation and PD-1 blockade leads to improved therapeutic efficacy and prolongs survival in murine models when compared to monotherapy. Overall, the findings elucidate the mechanisms of immune evasion in anti-tumor immunotherapies and suggest a potential combinatorial treatment strategy in RCC.

Aerobic granular sludge formation and stability in enhanced biological phosphorus removal system under antibiotics pressure: Performance, granulation mechanism, and microbial successions.

Wastewater containing antibiotics can pose a significant threat to biological wastewater treatment processes. This study investigated the establishment and stable operation of enhanced biological phosphorus removal (EBPR) by aerobic granular sludge (AGS) under mixed stress conditions induced by tetracycline (TC), sulfamethoxazole (SMX), ofloxacin (OFL), and roxithromycin (ROX). The results show that the AGS system was efficient in removing TP (98.0%), COD (96.1%), and NH4+-N (99.6%). The average removal efficiencies of the four antibiotics were 79.17% (TC), 70.86% (SMX), 25.73% (OFL), and 88.93% (ROX), respectively. The microorganisms in the AGS system secreted more polysaccharides, which contributed to the reactor's tolerance to antibiotics and facilitated granulation by enhancing the production of protein, particularly loosely bound protein. Illumina MiSeq sequencing revealed that putative phosphate accumulating organisms (PAOs)-related genera (Pseudomonas and Flavobacterium) were enormously beneficial to the mature AGS for TP removal. Based on the analysis of extracellular polymeric substances, extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory, and microbial community, a three-stage granulation mechanism was proposed including adaption to the stress environment, formation of early aggregates and maturation of PAOs enriched microbial granules. Overall, the study demonstrated the stability of EBPR-AGS under mixed antibiotics pressure, providing insight into the granulation mechanism and the potential use of AGS for wastewater treatment containing antibiotics.

[Combined Remediation of Eutrophic Water by Phoslock® and Aerobic Denitrifying Bacteria].

Nitrogen and phosphorus are the leading causes of water eutrophication, and it is challenging to remove nitrogen and phosphorus effectively through a single water remediation method. In this study, an aerobic denitrifying bacterium (AD-19) isolated from eutrophic water was used to construct an immobilized biofilm and combined with Phoslock® to remove nitrogen and phosphorus from the water. The phosphorus control efficiency of Phoslock®, nitrogen removal performance of the denitrifying bacteria, and combined remediation performance for the eutrophic water were studied. The results demonstrated that the removal rate of PO43--P in the simulated eutrophic water reached 95% with a dosing ratio of 80 (mass ratio of Phoslock® to PO43--P), and phosphorus release from sediment was effectively inhibited at the same time. Strain AD-19, which was identified as Pseudomonas sp. Using the 16S rDNA method, had a good heterotrophic nitrification and aerobic denitrification ability, and more than 97% of the nitrogen was removed when NH4+-N or NO3--N was used as the nitrogen source. The feasibility of the combined remediation of the eutrophic water was demonstrated using a lake simulation device. Furthermore, this technique was used to restore a eutrophic pond in a park in Wuhan city. After 16 days of treatment, the water quality indices for nitrogen and phosphorus were improved from worse than Grade Ⅴ to Grade Ⅲ (GB 3838-2002, Ministry of Environmental Protection of China, 2002) and remained stable for more than 270 days, indicating that Phoslock® combined with the immobilized biofilm could quickly and effectively restore eutrophic water as well as maintain the water quality for long periods.

Simultaneous removal of nitrogen and phosphorus by a novel aerobic denitrifying phosphorus-accumulating bacterium, Pseudomonas stutzeri ADP-19.

A novel denitrifying phosphorus-accumulating bacterium was isolated from contaminated sediment and identified as Pseudomonas stutzeri ADP-19. Bio-safety assays demonstrated that the strain was γ-hemolytic, antibiotic-sensitive, and had no decarboxylase activity. It removed 96.5% of NH4+-N and 73.3% of PO43--P (at initial concentrations of 100 mg/L and 20 mg/L) under aerobic conditions, and the corresponding maximum removal rates were 3.44 and 0.41 mg/L/h, respectively. Nitrogen removal was achieved through a fully nitrification-denitrification pathway [NH4+-N â†’ NH2OH â†’ NO2--N â†’ NO3--N â†’ NO2--N → (NxO) â†’ N2], while phosphorus removal mainly depended on the phosphate assimilation and the excessive poly-P accumulation. Strain ADP-19 also showed a strong salt tolerance within a wide salinity range of 0-5%. The enhanced biological treatment of anaerobic-digested wastewater in a sequencing batch reactor (SBR) indicated that the strain improved the microbial diversity of the activated sludge and significantly enhanced the nitrogen and phosphorus removal efficiency.

Early outcomes of thulium laser versus transurethral resection of the prostate for managing benign prostatic hyperplasia: a systematic review and meta-analysis of comparative studies.

PURPOSE: To assess the efficacy and safety of thulium laser resection of the prostate (TmLRP) vs transurethral resection of the prostate (TURP) for treating patients with benign prostatic hyperplasia (BPH). METHODS: A systematic search of the electronic databases, including Medline, Scopus, China National Knowledge Infrastructure, and the Cochrane Library was performed up to May 1, 2013. The pooled outcomes of interest assessing the two techniques included demographic and clinical baseline characteristics, perioperative variables, complications, and postoperative efficacy including maximum flow rate (Qmax), postvoid residual (PVR), quality of life (QoL) and International Prostate Symptom Score (IPSS). RESULTS: Nine trials assessing TmLRP vs. TURP were considered suitable for meta-analysis including three randomized controlled trials (RCTs), two prospective, and four retrospective studies. Compared with TURP, although TmLRP needed a longer operative time (weighted mean difference [WMD]: 9.00 min; 95% confidence interval [CI], 2.53-15.47; P=0.006), patients having TmLRP might benefit from significantly less serum sodium decreased (-3.58 mmol/L; 95% CI, -4.04 to -3.12; P<0.001), less serum hemoglobin decreased (WMD: -0.94 mmol/L; 95% CI, -1.44 to -0.44; P<0.001), shorter time of catheterization (WMD: -2.07 days; 95% CI, -2.66 to -1.49; P<0.001), shorter length of hospital stay (WMD: -1.87 days; 95% CI, -2.41 to -1.33; P<0.001), and fewer total complications (odds ratio [OR]: 0.29; 95% CI, 0.20-0.41; P<0.001). During the 1, 3, 6, and 12 months of postoperative follow-up, the procedures did not demonstrate a significant difference in Qmax, IPSS, PVR, and QoL. CONCLUSIONS: Our data suggest that as a promising minimally invasive technique, TmLRP appears to be a safe, feasible, and efficient alternative to TURP for treating patients with BPH with reliable perioperative safety, fewer complications, and comparable efficacy in relation to Qmax, PVR, QoL, and IPSS. Because of the inherent limitations of the included studies, further large sample prospective, multicentric, long-term follow-up studies and RCTs should be undertaken to confirm our findings.

Ammonium nitrogen removal in batch cultures treating digested piggery wastewater with microalgae Oedogonium sp.

Due to the nutrient characteristics of the high concentration of available ammonium in digested piggery wastewater (DPW), microalgae can be used to treat DPW before its final discharge. Four green microalgae (Hydrodictyaceae reticulatum Lag, Scenedesmus obliquus, Oedogonium sp. and Chlorella pyrenoidosa) and three blue-green algae (Anabaena flos-aquae, Oscillatoria amoena Gom and Spirulina platensis) were used to remove the nutrients (N, P, C), especially ammonium nitrogen (NH4(+)-N), from diluted DPW with 300 mg/L algae density in batch tests. The microalgae with the best NH4(+)-N nutrient removal was then selected for further optimization of the variables to improve NH4(+)-N removal efficiency using a central composite design (CCD) experiment. Taking into account the nutrient removal efficiency, Oedogonium sp. showed the best performance (reduction of 95.9% NH4(+)-N, 92.9% total phosphorus (TP) and 62.5% chemical oxygen demand (COD)) based on the results of the batch tests. The CCD results suggested that the optimal values of variables were initial Oedogonium sp. density of 399.2 mg/L and DPW diluted by 16.3, while the predicted value of NH4(+)-N removal efficiency obtained was 97.0%.

Generation and functional characterization of the anti-transferrin receptor single-chain antibody-GAL4 (TfRscFv-GAL4) fusion protein.

BACKGROUND: The development of vectors for cell-specific gene delivery is a major goal of gene therapeutic strategies. Transferrin receptor (TfR) is an endocytic receptor and identified as tumor relative specific due to its overexpression on most tumor cells or tissues, and TfR binds and intakes of transferrin-iron complex. We have previously generated an anti-TfR single-chain variable fragments of immunoglobulin (scFv) which were cloned from hybridoma cell line producing antibody against TfR linked with a 20 aa-long linker sequence (G4S)4. In the present study, the anti-TfR single-chain antibody (TfRscFv) was fused to DNA-binding domain of the yeast transcription factor GAL4. The recombinant fusion protein, designated as TfRscFv-GAL4, is expected to mediate the entry of DNA-protein complex into targeted tumor cells. RESULTS: Fusion protein TfRscFv-GAL4 was expressed in an E. coli bacterial expression system and was recovered from inclusion bodies with subsequent purification by metal-chelate chromatography. The resulting proteins were predominantly monomeric and, upon refolding, became a soluble biologically active bifunctional protein. In biological assays, the antigen-binding activity of the re-natured protein, TfRscFv-GAL4, was confirmed by specific binding to different cancer cells and tumor tissues. The cell binding rates, as indicated by flow cytometry (FCM) analysis, ranged from 54.11% to 8.23% in seven different human carcinoma cell lines. It showed similar affinity and binding potency as those of parent full-length mouse anti-TfR antibody. The positive binding rates to tumor tissues by tissue microarrays (TMA) assays were 75.32% and 63.25%, but it showed weakly binding with hepatic tissue in 5 cases, and normal tissues such as heart, spleen, adrenal cortex blood vessel and stomach. In addition, the re-natured fusion protein TfRscFv-GAL4 was used in an ELISA with rabbit anti-GAL4 antibody. The GAL4-DNA functional assay through the GAL4 complementary conjugation with the GAL4rec-GFP-pGes plasmid to verify the GLA4 activity and GAL4rec-recognized specificity functions. It also shows the complex, TfRscFv-GAL4-GAL4rec-GFP-pGes, could be taken into endochylema to express the green fluorescent protein (GFP) with 8 to 10-fold transfection efficiency. CONCLUSIONS: Results of our study demonstrated that the biofunctianality of genetically engineered fusion protein, TfRscFv-GAL4, was retained, as the fusion protein could both carry the plasmid of GAL4rec-pGes and bind TfR on tumour cells. This product was able to transfect target cells effectively in an immuno-specific manner, resulting in transient gene expression. This protein that can be applied as an effective therapeutic and diagnostic delivery to the tumor using endogenous membrane transport system with potential widespread utility.

Allothermal gasification of biomass using micron size biomass as external heat source.

An allothermal biomass gasification system using biomass micron fuel (BMF) as external heat source was developed. In this system, heat supplied to gasifier was generated from combustion of BMF. Biomass feedstock was gasified with steam and then tar in the produced gas was decomposed in a catalytic bed with NiO/γ-Al(2)O(3) catalyst. Finally the production gas was employed as a substitute for civil fuel gas. An overall energy analysis of the system was also investigated. The results showed that the lower heating value of the product gas reached more than 12 MJ/Nm(3). The combusted BMF accounted for 26.8% of the total energy input. Allothermal gasification based on the substituted BMF for conventional energy was an efficient and economical technology to obtain bioenergy.

VEGF pathway-targeted therapy for advanced renal cell carcinoma: a meta-analysis of randomized controlled trials.

Immunotherapy which has been in practice for more than 20 years proves effective for the treatment of metastatic renal cell carcinoma (mRCC). Anti-angiogenesis-targeted therapy has recently been identified as a promising therapeutic strategy for mRCC. This study was aimed to evaluate the effectiveness of vascular endothelial growth factor (VEGF) pathway-targeted therapy for mRCC by comparing its effectiveness with that of immunotherapy. The electronic databases were searched. Randomized controlled trials (RCTs) on comparison of VEGF inhibiting drugs (sorafenib, sunitinib and bevacizumab) with interferon (IFN) or placebo for mRCC treatment were included. Data were pooled to meta-analyze. A total of 7 RCTs with 3451 patients were involved. The results showed that anti-VEGF agents improved progression-free survival (PFS) and offered substantial clinical benefits to patients with mRCC. Among them, sunitinib had a higher overall response rate (ORR) than IFN (47% versus 12%, P<0.000001). Bevacizumab plus IFN produced a superior PFS [risk ratio (RR): 0.86, 95% confidence interval (CI): 0.76-0.97; P=0.01] and ORR (RR: 2.19; 95% CI: 1.72-2.78; P<0.00001) in patients with mRCC over IFN, but it yielded an increase by 31% in the risk of serious toxic effects (RR: 1.31; 95% CI: 1.20-1.43; P<0.00001) as compared with IFN. The overall survival (OS) was extended by sorafenib (17.8 months) and sunitinib (26.4 months) as compared with IFN (13 months). It was concluded that compared with IFN therapy, VEGF pathway-targeted therapies improved PFS and achieved significant therapeutic benefits in mRCC. However, the risk to benefit ratio of these agents needs to be further evaluated.