Comparison of Clinical Outcomes Between Modern Augmented ACL Repair and Autograft ACL Reconstruction: A Systematic Review and Meta-analysis of Studies With Minimum 2-Year Follow-up.

Background: Anterior cruciate ligament (ACL) reconstruction has been the gold standard for primary ACL rupture since the 1990s. In the past decade, ACL repair has received renewed attention and increased research. Purpose: To compare the clinical outcomes of modern augmented ACL repair versus autograft reconstruction for ACL ruptures. Study Design: Systematic review; Level of evidence, 3. Methods: A search of the PubMed, EMBASE, Scopus, Web of Science, and the Cochrane Library databases was conducted for prospective or retrospective comparative studies published between January 1, 2010, and January 3, 2023, with a minimum 2-year follow-up. Two independent reviewers performed data extraction and methodological quality assessment. Sensitivity analysis was performed to maintain the stability of results. Results: Nine studies were included (minimum follow-up period, 24-60 months). The total sample size was 833 patients (augmented repair group: 358 patients; autograft ACL reconstruction group: 475 patients). There were 4 randomized controlled trials (level 1), 1 prospective comparative study (level 2), 2 retrospective comparative studies (level 3), and 2 case-control studies (level 3). The augmented ACL repair group attained significantly higher Lysholm score (weighted mean difference [WMD] = 1.57; 95% confidence interval [CI], 0.14-3.01; P = .03) and hamstring strength (WMD = 36.69; 95% CI, 29.07-44.31; P < .01) but had higher rates of hardware removal (odds ratio [OR] = 6.30; 95% CI, 2.44-16.23; P = .0001), reoperation (OR = 1.87; 95% CI, 1.33-2.62; P = .0003), and failure (OR = 1.58; 95% CI, 1.03-2.43; P = .0003) compared with the autograft ACL reconstruction group. No significant differences were observed between the repair and reconstruction groups regarding postoperative International Knee Documentation Committee scores, Tegner scores, knee laxity, satisfaction, ACL revisions, complications, and reoperation rather than revision. Conclusion: Augmented ACL repair was associated with higher rates of reoperation, hardware removal, and failure compared with autograft ACL reconstruction in studies with minimum 2-year follow-up data. However, augmented ACL repair had higher Lysholm scores and hamstring strength versus autograft ACL reconstruction.

Characteristics and survival of primary urothelial carcinoma of the prostate: A multi-center retrospective study of 18 cases.

OBJECTIVES: To explore the features, treatment, and outcomes of primary urothelial carcinoma of the prostate (PUCP) in a multicenter study. METHODS: The clinical and imaging features, pathological findings, treatment, and outcomes of patients diagnosed with PUCP from January 2011 to April 2022 at three institutions were collected and analyzed. The Kaplan-Meier method and log-rank test were used to assess survival rates of the overall group and survival differences between groups according to TNM stage. RESULTS: The study cohort comprised 18 patients with PUCP of mean age 72.4±7.8 years. Dysuria and urinary frequency were the most common symptoms (77.8 %). Sixteen (88.9 %) patients had normal serum total PSA concentrations. Most patients showed abnormalities on urinalysis. MRI was the most accurate diagnostic imaging method (88.9 %). As to immunohistochemistry findings, GATA-3 (81.8 %) and P63 (84.6 %) were positive in most examined patients; however, no lesions were positive for PSA. Three (17.6 %) patients with T1N0M0 and T2N0M0 tumors underwent radical cystectomy. Eleven (64.7 %) patients which almost all had T4 tumors received systematic therapy, most of them receiving chemotherapy with gemcitabine and cisplatin, and radiotherapy. The median overall survival was 42 months, and the median progression-free survival 25 months, the latter being significantly longer in patients with T1-2 than in those with T3-4 disease (p=0.035). CONCLUSION: PUCP, a rare but highly aggressive type of prostate cancer, should be considered in men with abnormalities on MRI and normal serum PSA concentrations. Positive GATA-3, P63, and negative PSA are typical immunohistochemistry features. Radical cystectomy and systematic therapies can be effective.

Impact of BMI on the Survival of Renal Cell Carcinoma Patients Treated with Targeted Therapy: A Systematic Review and Meta-Analysis.

It is unclear whether obese renal cell carcinoma (RCC) patients treated with targeted therapy have better survival. We conducted this meta-analysis to assess the prognostic significance of body mass index (BMI) in RCC patients treated with targeted therapy. We systematically searched PubMed, Embase, Cochrane Library, and Web of Science by November 17, 2021. We calculated effect outcomes using random-effects and fixed-effects models. Fifteen articles were identified. We found that RCC patients treated with targeted therapy with BMI over 25 obtained better overall survival (OS) (hazard ratio [HR] = 0.69, 95% confidence interval [CI] = 0.58-0.82, I2 = 75.5%, p < 0.001) and progression-free survival (PFS) (HR = 0.71, 95%CI = 0.55-0.92, I2 = 69.7%, p = 0.006) than patients with BMI below 25. Obese (BMI over 30) patients had remarkably better OS (HR = 0.77, 95%CI = 0.70-0.85, I2 = 0.0%, p = 0.439) and PFS (HR = 0.86, 95%CI = 0.77-0.97, I2 = 0.0%, p = 0.934) than patients with BMI below 25. Overweight (BMI over 25 but below 30) patients also had better OS (HR = 0.86, 95%CI = 0.79-0.93, I2 = 17.7%, p = 0.295) and PFS (HR = 0.82, 95%CI = 0.74-0.90, I2 = 0.0%, p = 0.904) than patients with BMI below 25. When using BMI as continuous variable, patients with high BMI also obtained significantly better OS (HR = 0.92, 95%CI = 0.88-0.96, I2 = 0.0%, p = 0.806). Therefore, higher BMI was associated with greater OS and PFS in RCC patients treated with targeted therapy.

Development and validation of a preoperative nomogram to predict lymph node metastasis in patients with bladder urothelial carcinoma.

PURPOSE: Predicting lymph node metastasis (LNM) in patients with bladder urothelial carcinoma (BUC) before radical cystectomy aids clinical decision making. Here, we aimed to develop and validate a nomogram to preoperatively predict LNM in BUC patients. METHODS: Patients with histologically confirmed BUC, who underwent radical cystectomy and bilateral lymphadenectomy, were retrospectively recruited from two institutions. Patients from one institution were enrolled in the primary cohort, while those from the other were enrolled in the external validation cohort. Patient demographic, pathological (using transurethral resection of the bladder tumor specimens), imaging, and laboratory data were recorded. Univariate and multivariate logistic regression analyses were performed to explore the independent preoperative risk factors and develop the nomogram. Internal and external validation was conducted to assess nomogram performance. RESULTS: 522 and 215 BUC patients were enrolled in the primary and external validation cohorts, respectively. We identified tumor grade, infiltration, extravesical invasion, LNM on imaging, tumor size, and serum creatinine levels as independent preoperative risk factors, which were subsequently used to develop the nomogram. The nomogram showed a good predictive accuracy, with area under the receiver operator characteristic curve values of 0.817 and 0.825 for the primary and external validation cohorts, respectively. The corrected C-indexes, calibration curves (after 1000 bootstrap resampling), decision curve analysis results, and clinical impact curves demonstrated that the nomogram performed well in both cohorts and was highly clinically applicable. CONCLUSION: We developed a nomogram to preoperatively predict LNM in BUC, which was highly accurate, reliable, and clinically applicable.

Localization on a-priori information of plane extraction.

Localization constitutes a critical challenge for autonomous mobile robots, with flattened walls serving as a fundamental reference for indoor localization. In numerous scenarios, prior knowledge of a wall's surface plane is available, such as planes in building information modeling (BIM) systems. This article presents a localization technique based on a-priori plane point cloud extraction. The position and pose of the mobile robot are estimated through real-time multi-plane constraints. An extended image coordinate system is proposed to represent any planes in space and establish correspondences between visible planes and those in the world coordinate system. Potentially visible points representing the constrained plane in the real-time point cloud are filtered using the filter region of interest (ROI), derived from the theoretical visible plane region within the extended image coordinate system. The number of points representing the plane influences the calculation weight in the multi-plane localization approach. Experimental validation of the proposed localization method demonstrates its allowance for redundancy in initial position and pose error.

Granulomatous prostatitis after bacille Calmette-Guérin instillation resembles prostate carcinoma: A case report and review of the literature.

BACKGROUND: Bacille Calmette-Guérin (BCG) instillation is recommended in patients with non-muscle-invasive bladder cancer who have intermediate-risk and high-risk tumors. However, granulomatous prostatitis is a rare complication induced by BCG instillation, which can easily be misdiagnosed as prostate cancer. Here, we report a case of granulomatous prostatitis that resembled prostate cancer. CASE SUMMARY: A 64-year-old Chinese man with bladder cancer received BCG instillation. Three days later, he stopped BCG instillation and received anti-infective therapy due to the urinary tract infection. Three months after BCG restart, he had rising total prostate-specific antigen (PSA) (9.14 ng/mL) and decreasing free PSA/total PSA (0.09). T2-weighted images of magnetic resonance imaging (MRI) showed a 28 mm × 20 mm diffuse low signal abnormality in the right peripheral zone, which was markedly hyperintense on high b-value diffusion-weighted MRI and hypointense on apparent diffusion coefficient map images. Considering Prostate Imaging Reporting and Data System score of 5 and possibility of prostate cancer, a prostate biopsy was conducted. Histopathology showed typical features of granulomatous prostatitis. The nucleic acid test for tuberculosis was positive. He was finally diagnosed with BCG-induced granulomatous prostatitis. Thereafter, he stopped BCG instillation and received anti-tuberculosis treatment. During 10 mo follow-up, he had no evidence of tumor recurrence or symptoms of tuberculosis. CONCLUSION: Temporarily elevated PSA and high followed by low signal abnormality on diffusion-weighted MRI are important indicators of BCG-induced granulomatous prostatitis.

Precision-Driven Multi-Target Path Planning and Fine Position Error Estimation on a Dual-Movement-Mode Mobile Robot Using a Three-Parameter Error Model.

The multi-target path planning problem is a universal problem to mobile robots and mobile manipulators. The two movement modes of forward movement and rotation are universally implemented in integrated, commercially accessible mobile platforms used in logistics robots, construction robots, etc. Localization error in multi-target path tracking is one of the crucial measures in mobile robot applications. In this article, a precision-driven multi-target path planning is first proposed. According to the path's odometry error evaluation function, the precision-optimized path can be discovered. Then, a three-parameter odometry error model is proposed based on the dual movement mode. The error model describes localization errors in terms of the theoretical motion command values issued to the mobile robot, the forward moving distances, and the rotation angles. It appears that the three error parameters follow the normal distribution. The error model is finally validated using a mobile robot prototype. The error parameters can be identified by analyzing the actual moving trajectory of arbitrary movements. The experimental localization error is compared to the simulated localization error in order to validate the proposed error model and the precision-driven path planning method. The OptiTrack motion capture device was used to capture the prototype mobile robot's pose and position data.

GHSR1a deficiency suppresses inhibitory drive on dCA1 pyramidal neurons and contributes to memory reinforcement.

Growth hormone secretagogue receptor 1a (GHSR1a)-the receptor for orexigenic hormone ghrelin-is a G protein-coupled receptor that is widely distributed in the brain, including the hippocampus. Studies have demonstrated that genetic deletion of GHSR1a affects memory, suggesting the importance of ghrelin/GHSR1a signaling in cognitive control. However, current reports are controversial, and the mechanism underlying GHSR1a modulation of memory is uncertain. Here, we first report that global GHSR1a knockout enhances hippocampus-dependent memory, facilitates initial LTP in dorsal hippocampal Schaffer Collateral-CA1 synapses, and downregulates Akt activity in the hippocampus. Moreover, we show that the intrinsic excitability of GAD67+ interneurons-rather than neighboring pyramidal neurons in the dCA1-is suppressed by GHSR1a deletion, an effect that is antagonized by acute application of the Akt activator SC79. In addition, the inhibitory postsynaptic currents (IPSCs) on dCA1 pyramidal neurons are selectively reduced in mice with a GHSR1a deficiency. Finally, we demonstrate that selectively increasing the excitability of parvalbumin-expressing interneurons by hM3Dq-DREADDs increases IPSCs on dCA1 pyramidal neurons and normalizes memory in Ghsr1a KO mice. Our findings thus reveal a novel mechanism underlying memory enhancement of GHSR1a deficiency and herein support an adverse effect of GHSR1a signaling in hippocampus-dependent memory processes.

Testis and epididymis-unusual sites of metastatic gastric cancer: A case report and review of the literature.

BACKGROUND: Although gastric cancer is one of the most prevalent cancers worldwide, cases of gastric cancer metastasis to the male reproductive system are rare. Here, we report a case involving testicular and epididymal gastric cancer metastases. CASE SUMMARY: A 75-year-old Chinese man complained of experiencing a palpable painful mass in the right scrotum for 6 mo. He had undergone distal gastrectomy with chemotherapy for pT3N3aMx poorly differentiated gastric adenocarcinoma 9 mo before. Physical examination revealed a moderate right hydrocele and a painful mass in the right testis and epididymis. Serum tumor biomarkers were all normal except for elevated beta-human chorionic gonadotropin levels. Computed tomography urography and B-ultrasound imaging revealed a moderate right hydrocele and a mixed solid-cystic mass in the right testicular and epididymal area. Thus, the patient underwent right radical orchiectomy. Immunohistochemical analysis revealed that the tumor cells were positive for pancytokeratins and caudal related homeodomain transcription 2. Metastatic, poorly differentiated gastric adenocarcinoma of the testis and epididymis was confirmed by pathology. He continued to undergo chemotherapy at the department of oncology of our hospital. Mesenteric lymph node metastases were found at the postoperative 1-mo follow-up. CONCLUSION: Palpable, painful scrotal mass, history of gastric cancer, and imaging features may indicate testicular and epididymal metastatic gastric cancer.

Bioaugmentation for low C/N ratio wastewater treatment by combining endogenous partial denitrification (EPD) and denitrifying phosphorous removal (DPR) in the continuous A2/O - MBBR system.

Endogenous partial denitrification (EPD) and denitrifying phosphorous removal (DPR) were combined in a novel A2/O - MBBR (Anaerobic Anoxic Oxic - Moving Bed Biofilm Reactor) system for low carbon/nitrogen (C/N) ratio wastewater treatment. The DPR performance was compared and the nutrient metabolism was elucidated based on the optimization of hydraulic retention time (HRT, 4-12 h) and nitrate recycling (R, 200%-600%). In the continuous-flow, the nitrate (NO3-) denitrification accompanied by nitrite (NO2-, via EPD) accumulation with the nitrate-to-nitrite transformation ratio (NTR) of 35.87%-43.31% in the anoxic zones. At HRT of 12 h with R of 500%, batch test initially revealed the DPR mechanism using both NO3- and NO2- as electron acceptor, where denitrifying phosphorus accumulation organisms (DPAOs) and denitrifying glycogen accumulation organisms (DGAOs) were the main contributors for EPD with incomplete denitrification (NO3- → NO2-). Furthermore, stoichiometry-based functional bacteria analysis displayed that higher bioactivity of DPAOs (NO2-→N2, 57.30%; NO3-→N2, 35.85%) over DGAOs (NO3-→N2, 6.85%) facilitated the anoxic NO3- reduction. Microbial community analysis suggested that Cluster I of Defluviicoccus-GAO group (∼4%) was responsible for stable NO2- accumulation performance via EPD, while increased Accumulibacter-PAO group (by ∼15%) contributed to the advanced nutrient removal. Based on the achievement of NO2- accumulation, the application feasibility of integrated EPD - DPR - Anammox for deep-level nutrient removal was discussed.

Elucidating nitrifying performance, nitrite accumulation and microbial community in a three-stage plug flow moving bed biofilm reactor (PF - MBBR).

A three-stage plug flow moving bed biofilm reactor (PF - MBBR, consisting of three identical chambers of N1, N2 and N3) was proposed for nitrifier enrichment using synthetic wastewater. During the stable operation, the average NH4+-N effluent was 0.67 mg/L and NH4+-N removal was as high as 97.19% with the nitrite accumulation ratio (NAR) of 54.23%, although the biofilm thickness and biomass both presented downward trends from N1 (296 µm, 2280 mg/L), N2 (248 µm, 1850 mg/L) to N3 (198 µm, 1545 mg/L). Particularly, the comparative results of three stages revealed that N2 showed the optimum NH4+-N removal (77.27%) and NAR (75.21%) in the continuous-flow, while NAR of N3 unexpectedly maintained a high level of 65.83% in the batch test, suggesting that ammonia oxidizing bacteria (AOB) accounted for absolute advantage over nitrite oxidizing bacteria (NOB). High-throughput sequencing initially verified different distribution of bacterial community structure, where N2 was far away from N1 and N3 with the lowest community richness and community diversity (operational taxonomic units (OTUs): 454(N2)<527(N3)<621(N1)). Proteobacteria (77.60%-83.09%), Bacteroidetes (1.66%-3.66%), Acidobacteria (2.28%-4.67%), and Planctomycetes (1.19%-6.63%) were the major phyla. At the genus level, AOB (mainly Nitrosomonas) accounted for 5.08% (N1), 20.74% (N2) and 14.24% (N3) while NOB (mainly Nitrospira) increased from 0.14% (N1), 7.06% (N2) to 4.91% (N3) with the total percentages of 5.22%, 27.80% and 19.15%. Finally, the application feasibility of MBBR optimization linked with nitrite (NO2--N) accumulation for deep-level nutrient removal was discussed.

Efficacy and adverse reactions of intra-arterial chemotherapy in patients with bladder cancer: A systematic review and meta-analysis.

This meta-analysis investigated the efficacies of intra-arterial chemotherapy (IAC) plus intravesical chemotherapy (IVC) versus IVC alone in patients with non-muscle-invasive bladder cancer (NMIBC), and preoperative IAC versus preoperative intravenous chemotherapy (IV) in patients with bladder cancer. We also assessed the adverse reactions (ARs) of IAC. We searched the PubMed, Embase, Cochrane Library, and Web of Science databases for English articles published before April 2021. The qualities of cohort studies and randomized controlled trials were analyzed using the Newcastle-Ottawa Scale and Cochrane risk-of-bias tool, respectively. Effect outcomes were computed by random-effects and fixed-effects models. Statistical analyses were conducted using Stata 16.0 and RevMan v5.3.0. A total of seven articles were included. The analysis revealed that IAC plus IVC significantly prolonged recurrence-free survival (RFS) (hazard ratio [HR] = 0.55, 95% confidence interval [CI] = 0.40-0.76, I2 = 0%) and progression-free survival (PFS) (HR = 0.59, 95% CI = 0.37-0.97, I2 = 0%) compared with IVC alone in NMIBC patients after transurethral resection of bladder tumor (TURBT), but had no effect on overall survival (OS), tumor recurrence interval, or tumor-specific death rate. Preoperative IAC had no significant OS benefit compared with preoperative IV in bladder cancer patients. Regarding ARs, patients treated with IAC were significantly more likely to develop grade 1-2 ARs, including nausea/vomiting (odds ratio [OR] = 26.38, 95% CI = 1.88-370.79, I2 = 78%), neutropenia (OR = 10.15, 95% CI = 3.01-34.24, I2 = 0%), hypoleukemia (OR = 5.49, 95% CI = 1.38-21.82, I2 = 26%), and increased alanine aminotransferase (OR = 12.28, 95% CI = 2.24-67.43, I2 = 0%), but there was no significant difference between grade 1-2 ARs and grade 3-4 ARs in terms of increased creatinine in patients treated with IAC. Therefore, administration of IAC plus IVC after TURBT improved RFS and PFS compared with IAC alone in patients with NMIBC. IAC was associated with mild ARs and was well tolerated by most patients.

Multi-omics integrative analysis and survival risk model construction of non-small cell lung cancer based on The Cancer Genome Atlas datasets.

Lung cancer is a major cause of cancer-associated mortality worldwide. However, the association between multi-omics data and survival in lung cancer is not fully understood. The present study investigated the performance of the methylation survival risk model in multi-platform integrative molecular subtypes and aimed to identify copy number (CN) variations and mutations that are associated with survival risk. The present study analyzed 439 lung adenocarcinoma cases based on DNA methylation, RNA, microRNA (miRNA), DNA copy number and mutations from The Cancer Genome Atlas datasets. First, six cancer subtypes were identified using integrating DNA methylation, RNA, miRNA and DNA copy number data. The least absolute shrinkage and selection operator (LASSO) regression algorithm was used to extract methylation sites of survival model and calculate the methylation-based survival risk indices for all patients. Survival for patients in the high-risk group was significantly lower compared with that for patients in the low-risk group (P<0.05). The present study also assessed methylation-based survival risks of the six subtypes and analyzed the association between survival risk and non-silent mutation rate, number of segments, fraction of segments altered, aneuploidy score, number of segments with loss of heterozygosity (LOH), fraction of segments with LOH and homologous repair deficiency. Finally, the specific copy number regions and mutant genes associated with the different subtypes were identified (P<0.01). Chromosome regions 17q24.3 and 11p15.5 were identified as those with the most survival risk-associated copy number variation regions, while a total of 29 mutant genes were significantly associated with survival (P<0.01).

Angelica Essential Oil Loaded Electrospun Gelatin Nanofibers for Active Food Packaging Application.

The development of food packaging possessing bioactivities which could extend the shelf life of food has gained increased interest in recent years. In this study, gelatin nanofibers with encapsulated angelica essential oil (AEO) were fabricated via electrospinning. The morphology of gelatin/AEO nanofibers was examined by scanning electron microscopy (SEM) and the addition of AEO resulted in the increase of fiber diameter. The proton nuclear magnetic resonance (1H-NMR) spectra were measured to confirm the presence of AEO in nanofibers. The hydrophobic property of gelatin nanofibers was also found to be improved with the addition of AEO. The nanofibers incorporated with AEO showed significant antioxidant activity and inhibitory effect against both Gram-negative and Gram-positive bacteria in a concentration dependent manner. Furthermore, the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay demonstrated that the developed gelatin/AEO nanofibers revealed no cytotoxicity effect. Thus, gelatin nanofibers incorporated with AEO can be used as potential food packaging.

3D-QSAR and Molecular Docking Studies on Design Anti-Prostate Cancer Curcumin Analogues.

BACKGROUND: Prostate cancer is one of the most common tumors in the world and the fifth leading cause of male cancer death. Although the treatment of localized androgen-dependent prostate cancer has been successful, the efficacy of androgen-independent metastatic disease is limited. Curcumin, a natural product, has been found to inhibit the proliferation of prostate cancer cells. OBJECTIVE: To design curcumin analogs with higher biological activity and lower toxicity and side effects for the treatment of prostate cancer. METHODS: In this study, the three dimensional-quantitative structure activity relationship (3DQSAR) and molecular docking studies were performed on 34 curcumin analogs as anti-prostate cancer compounds. We introduced OSIRIS Property Explorer to predict drug-related properties of newly designed compounds. RESULTS: The optimum CoMSIA model exhibited statistically significant results: the cross-validated correlation coefficient q2 is 0.540 and non-cross-validated R2 value is 0.984. The external predictive correlation coefficient Rext 2 is 0.792. The information of structure-activity relationship can be obtained from the CoMSIA contour maps. In addition, the molecular docking study of the compounds for 3ZK6 as the protein target revealed important interactions between active compounds and amino acids. CONCLUSION: Compound 28i may be a new type of anti-prostate cancer drug with higher biological activity and more promising development.

Electrospun Gelatin Nanofibers Encapsulated with Peppermint and Chamomile Essential Oils as Potential Edible Packaging.

Natural and edible materials have attracted increasing attention in food packaging, which could overcome the serious environmental issues caused by conventional non-biodegradable synthetic packaging. In this work, gelatin nanofibers incorporated with two kinds of essential oil (EO), peppermint essential oil (PO) and chamomile essential oil (CO), were fabricated by electrospinning for potential edible packaging application. Electron microscopy showed that smooth and uniform morphology of the gelatin/EOs was obtained, and the diameter of nanofibers was mostly enlarged with the increase of the EO content. The proton nuclear magnetic resonance spectrum confirmed the existence of PO and CO in nanofibers after electrospinning. The addition of EOs led to an enhancement of the water contact angle of nanofibers. The antioxidant activity was significantly improved for the nanofibers loaded with CO, while the antibacteria activity against Escherichia coli and Staphylococcus aureus was better for the fibers with PO addition. The combination of half PO and half CO in nanofibers compensated for their respective limitations and exhibited optimum bioactivities. Finally, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay with NIH-3T3 fibroblasts demonstrated the absence of cytotoxicity of the gelatin/EO nanofibers. Thus, our studies suggest that the developed gelatin/PO/CO nanofiber could be a promising candidate for edible packaging.

Stereospecificity of Enoylreductase Domains from Modular Polyketide Synthases.

An enoylreductase (ER) domain of a polyketide synthase module recruiting a methylmalonate extender unit sets the C2 methyl branch to either the S or R configuration during processing of a polyketide intermediate carried by an acyl carrier protein (ACP) domain. In the present study, pantetheine- and ACP-bound trans-2-methylcrotonyl substrate surrogates were used to scrutinize the stereospecificity of the ER domains. The pantetheine-bound thioester was reduced to mixtures of both 2 R and 2 S products, whereas the expected 2 S epimer was almost exclusively generated when the cognate ACP-bound substrate surrogate was utilized. The analogous incubation of an ER with the substrate surrogate carried by a noncognate ACP significantly increased the generation of the unexpected 2 R epimer, highlighting the dependence of stereospecificity on proper protein-protein interactions between ER and ACP domains. The ER mutant assays revealed the involvement of the conserved tyrosine and lysine in stereocontrol. Taken together, these results expand the current understanding of the ER stereochemistry and help in the engineering of modular PKSs.

[Heterologous expression and characterization of a thermostable acylCoA synthetase].

Objective: An acyl-CoA synthetase (ACS) was characterized by in vitro biochemical assays to get a potential biocatalyst for the generation of polyketide precursors such as propionyl-CoA and butyryl-CoA. Methods: An ACS was identified in Caldicellulosiruptor owensensis OL genome by BLAST program using malonyl-CoA synthetase as the query sequence. The enzyme was heterologously expressed in Escherichia coli cells and purified to homogeneity by affinity purification. In vitro biochemical assays were used to reveal the substrate profile, the optimum reaction condition, the stability and the kinetics of the enzyme while the site-directed mutagenesis was used to assay the function of active site residues. Results: The enzyme was promiscuous enough to accept propionic acid, butyric acid, 2-methyl-propionic acid, pentanoic acid, 2-methyl-butyric acid, 3-methyl-butyric acid and cyclohexanecarboxylic acid as the substrates. It was most active at 30℃ and pH 7.0 and relatively stable because 45% activity remained after 8 hours incubation at 70℃. Interestingly, the substrate specificity of the enzyme could be switched by engineering three residues of active site. Conclusion: The ACS from C. owensensis OL was a potential biocatalyst for the generation of polyketide precursors.

ScbR- and ScbR2-mediated signal transduction networks coordinate complex physiological responses in Streptomyces coelicolor.

In model organism Streptomyces coelicolor, γ-butyrolactones (GBLs) and antibiotics were recognized as signalling molecules playing fundamental roles in intra- and interspecies communications. To dissect the GBL and antibiotic signalling networks systematically, the in vivo targets of their respective receptors ScbR and ScbR2 were identified on a genome scale by ChIP-seq. These identified targets encompass many that are known to play important roles in diverse cellular processes (e.g. gap1, pyk2, afsK, nagE2, cdaR, cprA, cprB, absA1, actII-orf4, redZ, atrA, rpsL and sigR), and they formed regulatory cascades, sub-networks and feedforward loops to elaborately control key metabolite processes, including primary and secondary metabolism, morphological differentiation and stress response. Moreover, interplay among ScbR, ScbR2 and other regulators revealed intricate cross talks between signalling pathways triggered by GBLs, antibiotics, nutrient availability and stress. Our work provides a global view on the specific responses that could be triggered by GBL and antibiotic signals in S. coelicolor, among which the main echo was the change of production profile of endogenous antibiotics and antibiotic signals manifested a role to enhance bacterial stress tolerance as well, shedding new light on GBL and antibiotic signalling networks widespread among streptomycetes.