Analysis of Risk Factors for Recurrence after Transurethral Resection of Bladder Tumor in Patients with Non-Muscle Invasive Bladder Cancer: 2-Year Follow-Up Outcomes.

INTRODUCTION: The aim was to investigate the risk factors for recurrence after transurethral resection of bladder tumor (TURBT) in patients with non-muscle invasive bladder cancer (NMIBC) and to provide a basis for clinical prevention of recurrence of NMIBC. METHODS: From January 2012 to December 2020, 592 patients with NMIBC who underwent TURBT attending the Second Affiliated Hospital of Xi'an Jiaotong University were retrospectively included in this study. Patients were divided into relapse and relapse-free groups according to whether relapse occurred within 2 years. Ultimately, 72 patients were included in the relapse group and 350 patients were included in the relapse-free group. Observation indicators included age, sex, smoking, underlying disease (hypertension, diabetes, coronary heart disease), two or more lesions, tumor size, hematuria, pathology grading (low, medium, high), staging (Ta, T1), muscular invasion in initial pathology, tumor base (sessile, pedunculated), use of intravesical drug (pirarubicin, bacillus Calmette-Guerin [BCG], mitomycin, hydroxycamptothecin, gemcitabine). RESULTS: In this study, the 2-year recurrence rate of NMIBC patients after TURBT was 17.06%. There were significant differences in comparison of pirarubicin, BCG, and mitomycin treatment between the two groups (p < 0.05). To avoid missing risk factors for recurrence, factors with p < 0.1 were analyzed. The results of univariate logistic regression analysis showed that NMIBC patients with BCG treatment (OR = 5.088, 95% CI = 1.444-17.73, p = 0.012), high pathology grading (OR = 0.415, 95% CI = 0.197-0.880, p = 0.023), T1 stage (OR = 2.097, 95% CI = 0.996-4.618, p = 0.059), mitomycin treatment (OR = 5.029, 95% CI = 1.149-21.77, p = 0.031), and pirarubicin treatment (OR = 1.794, 95% CI = 1.079-3.030, p = 0.024) had significantly higher risk of recurrence within 2 years after TURBT. The results of multivariate logistic regression analysis showed that NMIBC patients with high pathology grading (OR = 0.4030, 95% CI = 0.1702-0.8426, p = 0.0241), pirarubicin treatment (OR = 1.961, 95% CI = 1.159-3.348, p = 0.0125), and BCG treatment (OR = 6.201, 95% CI = 1.275-29.73, p = 0.0190) had significantly higher risk of recurrence within 2 years after TURBT. CONCLUSION: Our study highlights the importance of postoperative surveillance and individualized treatment for patients with NMIBC. Our findings show that high pathology grading, pirarubicin treatment, and BCG treatment are independent risk factors for recurrence after TURBT in patients with NMIBC. However, caution is warranted when interpreting our findings due to the small sample size and the need for further research to confirm the negative impact of mitomycin and BCG on recurrence rates.

qPTMplants: an integrative database of quantitative post-translational modifications in plants.

As a crucial molecular mechanism, post-translational modifications (PTMs) play critical roles in a wide range of biological processes in plants. Recent advances in mass spectrometry-based proteomic technologies have greatly accelerated the profiling and quantification of plant PTM events. Although several databases have been constructed to store plant PTM data, a resource including more plant species and more PTM types with quantitative dynamics still remains to be developed. In this paper, we present an integrative database of quantitative PTMs in plants named qPTMplants (http://qptmplants.omicsbio.info), which hosts 1 242 365 experimentally identified PTM events for 429 821 nonredundant sites on 123 551 proteins under 583 conditions for 23 PTM types in 43 plant species from 293 published studies, with 620 509 quantification events for 136 700 PTM sites on 55 361 proteins under 354 conditions. Moreover, the experimental details, such as conditions, samples, instruments and methods, were manually curated, while a variety of annotations, including the sequence and structural characteristics, were integrated into qPTMplants. Then, various search and browse functions were implemented to access the qPTMplants data in a user-friendly manner. Overall, we anticipate that the qPTMplants database will be a valuable resource for further research on PTMs in plants.

iCysMod: an integrative database for protein cysteine modifications in eukaryotes.

As important post-translational modifications, protein cysteine modifications (PCMs) occurring at cysteine thiol group play critical roles in the regulation of various biological processes in eukaryotes. Due to the rapid advancement of high-throughput proteomics technologies, a large number of PCM events have been identified but remain to be curated. Thus, an integrated resource of eukaryotic PCMs will be useful for the research community. In this work, we developed an integrative database for protein cysteine modifications in eukaryotes (iCysMod), which curated and hosted 108 030 PCM events for 85 747 experimentally identified sites on 31 483 proteins from 48 eukaryotes for 8 types of PCMs, including oxidation, S-nitrosylation (-SNO), S-glutathionylation (-SSG), disulfide formation (-SSR), S-sulfhydration (-SSH), S-sulfenylation (-SOH), S-sulfinylation (-SO2H) and S-palmitoylation (-S-palm). Then, browse and search options were provided for accessing the dataset, while various detailed information about the PCM events was well organized for visualization. With human dataset in iCysMod, the sequence features around the cysteine modification sites for each PCM type were analyzed, and the results indicated that various types of PCMs presented distinct sequence recognition preferences. Moreover, different PCMs can crosstalk with each other to synergistically orchestrate specific biological processes, and 37 841 PCM events involved in 119 types of PCM co-occurrences at the same cysteine residues were finally obtained. Taken together, we anticipate that the database of iCysMod would provide a useful resource for eukaryotic PCMs to facilitate related researches, while the online service is freely available at http://icysmod.omicsbio.info.

Effects of dietary supplementation of bovine lactoferrin on growth performance, immune function and intestinal health in weaning piglets.

Weaning is a crucial period in the pig's life cycle, which is frequently followed by gastrointestinal (GI) infections, diarrhea and even death. This study focused on the impact of bovine lactoferrin (bLF) supplementation on the intestinal health of weaning piglets. Weaning piglets (Duroc × Landrace × Yorkshire, 23 days) were randomly allocated into four groups, which included negative control group (CON): basic diet; positive control group (ANT): basic diet + 20 mg/kg flavomycin + 100 mg/kg aureomycin; treatment group bLF-A: basic diet + 1 g/kg bLF; treatment group bLF-B: basic diet + 3 g/kg bLF. The result showed that dietary supplementation of bLF can improve growth performance and reduce diarrhea, which exhibits dose-dependency (P < 0.05). Compared with CON group, supplementation with bLF significantly improved immunity, and increased villus height and ratio of villus height/crypt depth at the small intestinal mucosa (P < 0.05). The mRNA expression of claudin-1, occludin and ZO-1 was greatly increased in the ileum of bLF group on days 7 and 14 (P < 0.05). Furthermore, the supplementation of bLF increased the abundance of Lactobacillus and Bifidobacterium and decreased the abundance of Escherichia coli in the cecum on day 7 (P < 0.05). The dietary supplementation of bLF enhanced the growth performance, reduced diarrhea rate in weaning piglets by improving intestinal immunity, morphology and barrier function, balancing intestinal microbiota. And bLF can be a promising feed additive in relieving stress situation of weaning piglets.

Changes in gene expression and enzyme activity related to glucose metabolism in the livers of Brandt's voles (Lasiopodomys brandtii) exposed to hypoxia.

Brandt's vole (Lasiopodomys brandtii) is a hypoxia-tolerant species, and the metabolic characteristics of hypoxia-tolerant species have become a focus of recent research. However, insights into the anaerobic and aerobic metabolism of the livers of Brandt's voles under hypoxia remain limited. In this study, Brandt's voles and hypoxia-intolerant Kunming mice (Mus musculus, control species) were exposed to hypoxia conditions (Brandt's voles, 10% and 7.5% O2; Kunming mice, 10% O2) for 24 h, and changes in gene expression and enzyme activity related to anaerobic and aerobic metabolism in the livers were evaluated. Phosphofructokinase 1 (PFK1), phosphofructokinase 2 (PFK2), pyruvate kinase muscle (PKM), hexokinase 2 (HK2), and lactate dehydrogenase (LDH) related to anaerobic metabolism in the livers of Brandt's voles were increased under 7.5% O2. Regarding gene expression and enzyme activity for aerobic metabolism in Brandt's voles under 7.5% and 10% O2, pyruvate dehydrogenase kinase 1 (PDK1) expression was up-regulated, and succinate dehydrogenase (SDH) activity was decreased. In the livers of Kunming mice, gene expression related to anaerobic and aerobic metabolism was increased at the late stage of 10% O2, and SDH activity was enhanced at 6 h and reduced at 18 h. In addition, PFK1,PKM, PDK1 expression and SDH activity in Brandt's voles were significantly correlated with HIF-1a expression. PFK1, PKM, LDHand PDK1 expression in Kunming mice were significantly correlated with HIF-1a expression. These findings indicate that the livers of Brandt's voles have a certain tolerance to hypoxia, and metabolic changes play important roles in hypoxia tolerance.

Resistance response to Arenicin derivatives in Escherichia coli.

The rising prevalence of antibiotic resistance poses the greatest health threats. Antimicrobial peptides (AMPs) are regarded as the potentially effective therapy. To avoid current crisis of antibiotic resistance, a comprehensive understanding of AMP resistance is necessary before clinical application. In this study, the development of resistance to the anti-Gram-negative bacteria peptide N6NH2 (21 residues, ß-sheet) was characterized in E. coli ATCC25922. Three N6NH2-resistant E. coli mutants with 32-fold increase in MIC were isolated by serially passaging bacterial lineages in progressively increasing concentrations of N6NH2 and we mainly focus on the phenotype of N6NH2-resistant bacteria different from sensitive bacteria. The results showed that the resistance mechanism was attributed to synergy effect of multiple mechanisms: (i) increase biofilm formation capacity (3 ~ 4-fold); (ii) weaken the affinity of lipopolysaccharide (LPS) with N6NH2 (3 ~ 8-fold); and (iii) change the cell membrane permeability and potential. Interestingly, a chimeric peptide-G6, also a N6NH2 analog, which keep the same antibacterial activity to both wild-type and resistant clones (MIC value: 16 µg/mL), could curb N6NH2-resistant mutants by stronger inhibition of biofilm formation, stronger affinity with LPS, and stronger membrane permeability and depolarization than that of N6NH2.

The remarkable hypoxia tolerance in Brandt's voles (Lasiopodomys brandtii).

Oxygen (O2 ) is essential for the survival of most animal species; however, O2 levels are lower than normal in certain spatiotemporal circumstances. Therefore, animals have to develop some strategies to face and adapt to this plight. This study compared the differences in survival, behavior, and hippocampal neurons under hypoxic conditions between Brandt's voles (Lasiopodomys brandtii) and Kunming mice (Mus musculus). Brandt's voles had longer survival times than did Kunming mice in 5%-6.5% O2 levels. Significant differences were observed in mortality between Brandt's voles and Kunming mice after 6 hr of exposure to 5%-7% O2 levels. The novel object recognition test (NORT) and neuronal nuclei staining revealed the presence of species-dependent learning deficits and neuron loss under hypoxic conditions. NORT and neuronal nuclei staining showed that the Kunming mice exposed to 7.5% O2 levels had significant learning deficits and neuronal loss compared with normoxia Kunming mice. Conversely, Brandt's voles exposed to 7.5% O2 levels had no significant learning deficits and neuronal loss compared with the normoxia group. Therefore, hypoxia showed significant effects on survival, cognitive behavior, and neuronal loss of the two species. Moreover, Brandt's voles showed stronger hypoxia tolerance than Kunming mice, and Brandt's voles hypoxia tolerance may root in their adaptive evolution to highland habitat environment. This research is meaningful for studying hypoxic adaptation in animals, and will promote the development of therapies for some illnesses such as stroke and cardiac arrest.

The prognostic value of circulating tumour cells (CTCs) and CTC white blood cell clusters in patients with renal cell carcinoma.

PURPOSE: Circulating tumour cell (CTC) and CTC-white blood cell (CTC-WBC) clusters are related to the prognosis of tumour patients. However, the relationship between CTC-WBC clusters and prognosis in renal cell carcinoma (RCC) patients is not clear. We evaluated the prognostic value of CTC-WBC clusters using metastasis-free survival (MFS) and overall survival (OS) in patients with RCC. MATERIALS AND METHODS: The baseline, survival, and CTC data of patients with RCC were statistically analysed by R. RESULTS: The Cox risk proportional regression model suggests that the total CTCs, pathology type, and CTC-WBC clusters can be used as prognostic indicators for the MFS of RCC patients. Total CTCs and solid tumour diameter can be used as prognostic indicators for the OS of RCC patients. Using Kaplan-Meier survival analysis, we found that patients with total CTCs, pathology, and CTC-WBC clusters greater than the cut-off value had a worse MFS, and patients with total CTCs greater than the cut-off value had a worse OS. CONCLUSION: The analysis of the clinical sample data in patients with RCC shows that CTC-WBC clusters play an important role in monitoring the prognosis of RCC. Among them, total CTCs, pathology, and CTC-WBC clusters were combined as prognostic factors for the MFS of RCC patients. Total CTCs and solid tumour diameter can be combined as prognostic factors for the OS of RCC patients. These prognostic factors provide more convenient and accurate condition monitoring for renal cancer patients and can be used to actively improve the prognosis of patients.

Numerical Calculation of Apparent Contact Angles on the Hierarchical Surface with Array Microstructures by Wire Electrical Discharge Machining.

It is necessary to theoretically research wettability in superhydrophobic surface fabrication. Here, a numerical calculation approach is proposed for determining the contact angle of the water droplets on array micropillars by wire electrical discharge machining (WEDM). A hierarchical model is employed for these array microstructures, including mechanical analysis for a water droplet placed on a smooth array and wettability evaluation on the morphology of the WEDM surface. On pillars, equations are listed to solve the apparent contact angle according to force balance of gravity, tension, and pressure. As for the WEDM morphology, temperature simulation and measurement are carried out, and then the effect of roughness on surface wettability is studied. Constructed formulas predict the contact angle, and then the effect of geometric dimensions is obtained. In order to verify the assumption, array micropillars with different cross-profiles are prepared using high-speed WEDM on the Al alloy surface. Through the results of contact angle determination, the numerical calculation is carried out. This theoretical prediction is beneficial for improving the fabrication of the superhydrophobic surface by WEDM.

Potent intracellular antibacterial activity of a marine peptide-N6NH2 and its D-enantiomer against multidrug-resistant Aeromonas veronii.

Aeromonas veronii can cause a variety of diseases such as sepsis in humans and animals. However, there has been no effective way to eradicate A. veronii. In this study, the intracellular antibacterial activities of the C-terminal aminated marine peptide N6 (N6NH2) and its D-enantiomer (DN6NH2) against A. veronii were investigated in macrophages and in mice, respectively. The result showed that DN6NH2 with the minimum inhibitory concentration (MIC) of 1.62 µM is more resistant to cathepsin B than N6NH2 (3.23 µM). The penetration percentages of the cells treated with 4-200 µg/mL fluorescein isothiocyanate (FITC)-DN6NH2 were 52.5-99.6%, higher than those of FITC-N6NH2 (27.0-99.1%). Both N6NH2 and DN6NH2 entered macrophages by macropinocytosis and an energy-dependent manner. DN6NH2 reduced intracellular A. veronii by 34.57%, superior to N6NH2 (19.52%). After treatment with 100 µg/mL DN6NH2, the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1ß were reduced by 53.45%, 58.54%, and 44.62%, respectively, lower than those of N6NH2 (15.65%, 12.88%, and 14.10%, respectively); DN6NH2 increased the IL-10 level (42.94%), higher than N6NH2 (7.67%). In the mice peritonitis model, 5 µmol/kg DN6NH2 reduced intracellular A. veronii colonization by 73.22%, which was superior to N6NH2 (32.45%) or ciprofloxacin (45.67%). This suggests that DN6NH2 may be used as the candidate for treating intracellular multidrug-resistant (MDR) A. veronii. KEY POINTS: • DN6NH2 improved intracellular antibacterial activity against MDR A. veronii. • DN6NH2 entered macrophages by micropinocytosis and enhanced the internalization rates. • DN6NH2 effectively protected the mice from infection with A. veronii.

The Significance of Detection of Circulating Tumor Cells and Beclin1 in Peripheral Blood of Patients with Renal Cell Carcinoma.

AIM: In this study, the correlation between the circulating tumor cells (CTCs), clinical and pathological status, Beclin1 expression, and the prognosis of patients with renal cell carcinoma were studied. METHODS: The patients with renal cancer were tested every 3 months for 2 years, and once every 6 months after 2 years by using CanPatrol-ITMCTCs detection technology. The expression of Beclin1 in different types of CTCs was detected. The study investigated the correlation between Beclin1 expression of patients with different gender, age, tumor pathological stage, clinical stage, and postoperative metastasis. RESULTS: A total of 199 renal cancer patients were included in this study, and the patients underwent CTCs testing ranging from 1 to 10 times. There are 936 epithelial CTCs, 2,884 mixed CTCs, and 1,218 interstitial CTCs were detected. The results show that there are statistical differences between the three subtypes (P = 0.001). The cell count of the Beclin1 negative group was statistically significantly higher than that of the positive group among the three subtypes from the first to the fourth test (P < 0.05). The first test results showed that age was negatively correlated with the number of CTCs (r = -0.204, P = 0.004). There are no differences in the overall survival (OS) and disease-free survival (DFS) between the different numbers of CTCs and Beclin1 expression (all P values were > 0.05). CONCLUSION: The expression of Beclin1 in epithelial and mesenchymal renal cell carcinoma reduces the number of CTCs produced. The age of the patient may affect the levels of CTCs in renal cell carcinoma.

Transcriptome analysis of the response provided by Lasiopodomys mandarinus to severe hypoxia includes enhancing DNA repair and damage prevention.

BACKGROUND: Severe hypoxia induces a series of stress responses in mammals; however, subterranean rodents have evolved several adaptation mechanisms of energy metabolisms and O2 utilization for hypoxia. Mammalian brains show extreme aerobic metabolism. Following hypoxia exposure, mammals usually experience irreversible brain damage and can even develop serious diseases, such as hypoxic ischemic encephalopathy and brain edema. To investigate mechanisms underlying the responses of subterranean rodents to severe hypoxia, we performed a cross-species brain transcriptomic analysis using RNA sequencing and identified differentially expressed genes (DEGs) between the subterranean rodent Lasiopodomys mandarinus and its closely related aboveground species L. brandtii under severe hypoxia (5.0% O2, 6 h) and normoxia (20.9% O2, 6 h). RESULTS: We obtained 361 million clean reads, including 69,611 unigenes in L. mandarinus and 69,360 in L. brandtii. We identified 359 and 515 DEGs by comparing the hypoxic and normoxia groups of L. mandarinus and L. brandtii, respectively. Gene Ontology (GO) analysis showed that upregulated DEGs in both species displayed similar terms in response to severe hypoxia; the main difference is that GO terms of L. brandtii were enriched in the immune system. However, in the downregulated DEGs, GO terms of L. mandarinus were enriched in cell proliferation and protein transport and those of L. brandtii were enriched in nuclease and hydrolase activities, particularly in terms of developmental functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that upregulated DEGs in L. mandarinus were associated with DNA repair and damage prevention as well as angiogenesis and metastasis inhibition, whereas downregulated DEGs were associated with neuronal synaptic transmission and tumor-associated metabolic pathways. In L. brandtii, upregulated KEGG pathways were enriched in the immune, endocrine, and cardiovascular systems and particularly in cancer-related pathways, whereas downregulated DEGs were associated with environmental information processing and misregulation in cancers. CONCLUSIONS: L. mandarinus has evolved hypoxia adaptation by enhancing DNA repair, damage prevention, and augmenting sensing, whereas L. brandtii showed a higher risk of tumorigenesis and promoted innate immunity toward severe hypoxia. These results reveal the hypoxic mechanisms of L. mandarinus to severe hypoxia, which may provide research clues for hypoxic diseases.

Clinical application of drug sensitive gene detection in postoperative instillation for non-muscle invasive bladder cancer.

BACKGROUND: Bladder cancer is the most common malignant tumor of the urinary system. One of the biological characteristics of NMIBC is the high recurrence rate after surgery. The implementation of this project aimed to investigate the role of pharmacogenomic testing-guided intravesical perfusion of chemotherapeutic agents in the postoperative perfusion therapy for non-muscle invasive bladder cancer. METHOD: From January 2015 to December 2016, 298 patients with non-muscle-invasive bladder cancer were enrolled in this prospective study. These patients received chemotherapy drugs after electrotherapy. According to the presence or absence of tumor susceptibility gene detection after surgery, they were divided into two groups, including the drug sensitive group(N = 44) and the control group(N = 254). The drug sensitive group received bladder infusion therapy with sensitive chemotherapy drugs based on drug sensitivity gene detection results. The control group received intravesical instillation of pirarubicin. The preoperative general data and tumor grade of patients were recorded. Cystoscopy was performed before and every 3 months after surgery. The chest CT, upper abdomen CT, renal function, and urinary routine tests were performed. Tumor recurrence, metastasis and tumor-related death were recorded and evaluated during follow-up. RESULTS: The drug sensitive group, which selected high-sensitivity drugs for intravesical instillation therapy based on gene expression, has a significantly lower relapse rate (11.36% vs 37.40%, P < 0.05) and a significantly longer time to relapse (17.80 ± 7.20 month vs11.20 ± 6.10 month, P < 0.05) compared with the control group. There were no significant differences in the time of mortality and death time between two groups. CONCLUSION: The pharmacogenomic testing-directed bladder instillation of chemotherapeutic drugs may be more effective than empiric drug administration in reducing the recurrence rate of non-muscle-invasive bladder cancer.

Living near the limits: Effects of interannual variation in food availability on diet and reproduction in a temperate primate, the Taihangshan macaque (Macaca mulatta tcheliensis).

Nutrient intake of animals is influenced by an interplay of external and internal factors, such as food availability and reproductive state, respectively. We used the nutritional geometry framework to analyze individual-based data on energy and nutrient intake in relation to reproductive state in a population of rhesus macaques (Macaca mulatta tcheliensis), which live in a harsh high-latitude habitat, the Taihangshan mountains of China, and exhibit strong reproductive seasonality. We combined data over a 3-year period on food availability, diets, reproductive output, and components of maternal investment to understand how Taihangshan macaques respond to variation in food availability and nutrition in reproduction. Our results show there was high interannual variation in availability of an important staple source of fat and carbohydrates (nonprotein energy), seeds of oak (Quercus spp). Despite this variability in seed availability skewing the dietary macronutrient ratios considerably (from 12.96% to 30.12% dietary energy from protein), total metabolizable energy intake was maintained across years during pregnancy. Lactating females had higher mean daily energy intakes than pregnant females. As in pregnant females, energy intake was maintained constant across years, but only when seed availability enabled the contribution of available protein to energy intake to be maintained between 15.32% (2013) and 17.97% (2015). In 2014, when seeds were scarce, lactating females had a shortfall in energy intake compared with 2013 and 2015. This corresponded with a reduction in the number of females giving birth (11 out of 23), but there was no interannual difference in survival rates. Compared to 2013 and 2015, in 2014 females had greater weight loss (drew on body reserves), moved less, and spent more time nursing their offspring. We discuss implications of these results for range limitation in Taihangshan macaques.

Improved Stability and Activity of a Marine Peptide-N6NH2 against Edwardsiella tarda and Its Preliminary Application in Fish.

Edwardsiella tarda can cause fatal gastro-/extraintestinal diseases in fish and humans. Overuse of antibiotics has led to antibiotic resistance and contamination in the environment, which highlights the need to find new antimicrobial agents. In this study, the marine peptide-N6 was amidated at its C-terminus to generate N6NH2. The antibacterial activity of N6 and N6NH2 against E. tarda was evaluated in vitro and in vivo; their stability, toxicity and mode of action were also determined. Minimal inhibitory concentrations (MICs) of N6 and N6NH2 against E. tarda were 1.29-3.2 µM. Both N6 and N6NH2 killed bacteria by destroying the cell membrane of E. tarda and binding to lipopolysaccharide (LPS) and genomic DNA. In contrast with N6, N6NH2 improved the stability toward trypsin, reduced hemolysis (by 0.19% at a concentration of 256 µg/mL) and enhanced the ability to penetrate the bacterial outer and inner membrane. In the model of fish peritonitis caused by E. tarda, superior to norfloxacin, N6NH2 improved the survival rate of fish, reduced the bacterial load on the organs, alleviated the organ injury and regulated the immunity of the liver and kidney. These data suggest that the marine peptide N6NH2 may be a candidate for novel antimicrobial agents against E. tarda infections.

Dual Antibacterial Activities and Biofilm Eradication of a Marine Peptide-N6NH2 and Its Analogs against Multidrug-Resistant Aeromonas veronii.

Aeromonas veronii is one of the main pathogens causing various diseases in humans and animals. It is currently difficult to eradicate drug-resistant A. veronii due to the biofilm formation by conventional antibiotic treatments. In this study, a marine peptide-N6NH2 and its analogs were generated by introducing Orn or replacing with D-amino acids, Val and Pro; their enzymic stability and antibacterial/antibiofilm ability against multi-drug resistant (MDR) A. veronii ACCC61732 were detected in vitro and in vivo, respectively. The results showed that DN6NH2 more rapidly killed A. veronii ACCC61732 and had higher stability in trypsin, simulated gastric/intestinal fluid, proteinase K, and mouse serum than the parent peptide-N6NH2. DN6NH2 and other analogs significantly improved the ability of N6NH2 to penetrate the outer membrane of A. veronii ACCC61732. DN6NH2, N6PNH2 and V112N6NH2 protected mice from catheter-associated biofilm infection with MDR A. veronii ACCC61732, superior to N6NH2 and CIP. DN6NH2 had more potent efficacy at a dose of 5 µmol/kg (100% survival) in a mouse peritonitis model than other analogs (50-66.67%) and CIP (83.33%), and it inhibited the bacterial translocation, downregulated pro-inflammatory cytokines, upregulated the anti-inflammatory cytokine, and ameliorated multiple-organ injuries (including the liver, spleen, lung, and kidney). These data suggest that the analogs of N6NH2 may be a candidate for novel antimicrobial and antibiofilm agents against MDR A. veronii infections.

Microtopography-Guided Radial Gradient Circle Array Film with Nanoscale Resolution.

Distribution of multimaterials at arbitrary positions with nanoscale resolution and over a large area substrate is essential to future advances in functional graded materials. Such stringent requirements are highly beyond the reach of current techniques, although newly developed 3D printing technologies are addressed. Here, a radial gradient circle array film with the distribution accuracy up to ≈18 nm is fabricated by using microtopographic substrate. A mathematical model is developed to guide the distribution of position, size, shape, and type of materials on an arbitrary section for the given morphology of substrate. The periodic electrical and mechanical properties of the radial gradient circle film are identified, which can be beneficial for further functionalization and applications, such as gradient refractive index lenses, microcoils, and microantennas.

A recombinant fungal defensin-like peptide-P2 combats multidrug-resistant Staphylococcus aureus and biofilms.

There is an urgent need to discover new active drugs to combat methicillin-resistant Staphylococcus aureus, which is a serious threat to humans and animals and incompletely eliminated by antibiotics due to its intracellular accumulation in host cells, production of biofilms, and persisters. Fungal defensin-like peptides (DLPs) are emerging as a potential source of new antibacterial drugs due to their potent antibacterial activity. In this study, nine novel fungal DLPs were firstly identified by querying against UniProt databases and expressed in Pichia pastoris, and their antibacterial and anti-biofilm ability were tested against multidrug-resistant (MDR) S. aureus. Results showed that among them, P2, the highest activity and expression level, showed low toxicity, no resistance, and high stability. Minimal inhibitory concentrations (MICs) of P2 against Gram-positive bacteria were < 2 µg/mL. P2 exhibited the potent activity against intracellular MDR S. aureus (bacterial reduction in 80-97%) in RAW264.7 macrophages. P2 bound to/disrupted bacterial DNA, wrinkled outer membranes and permeabilized cytoplasmic membranes, but maintained the integrity of bacterial cells. P2 inhibited/eradicated the biofilm and killed 99% persister bacteria, which were resistant to 100× MIC vancomycin. P2 upregulated the anti-inflammatory cytokine (IL-10) and downregulated pro-inflammatory cytokines (TNF-α/IL-1ß) and chemokine (MCP-1) levels in RAW 264.7 macrophages and in mice, respectively. Five milligram per kilogram P2 enhanced the survival of S. aureus-infected mice (100%), superior to vancomycin (30 mg/kg), inhibited the bacterial translocation, and alleviated multiple-organ injuries (liver, spleen, kidney, and lung). These data suggest that P2 may be a candidate for novel antimicrobial agents against MDR staphylococcal infections.

Dietary diversity of an ecological and macronutritional generalist primate in a harsh high-latitude habitat, the Taihangshan macaque (Macaca mulatta tcheliensis).

Recent advances in niche theory have stressed the importance of understanding dietary generalism at multiple levels, including the range of habitat and foods exploited by a species, foods exploited within populations, and patterns of nutrient intake. Here we apply this framework to examine the dietary strategy of the Macaca mulatta, a primate species that is second only to humans in their breadth of geographical distribution, and occupy diverse ecological habitats from cold temperate to tropical latitudes. A recent study showed that the Taihangshan subspecies ( M. mulatta tcheliensis) in China, which is found at the northern latitudinal limit of the species range, respond to ecologically constrained interannual variation in the macronutrient ratios of the spring diet in a way that theory predicts should be associated with ecological generalism. Here we further extend this study, examining the relationships between seasonal variation in food availability and the patterns of food selection across a full year. We found that, despite the ecological and macronutritional generalism of the species, Taihangshan macaques subsist on a relatively small range of foods (57 different foods, spread across 8 categories comprising seeds, fruits, buds, flowers, leaves, herbs, young bark, and twigs), but face considerable seasonal variation in the combinations available. In spring and summer, when seeds were scarce, leaves accounted for 60.3 ± 13.8% of their diet (dry matter [DM] %), and herbs contributed 31.7 ± 22.2%. However, in autumn and winter, when seeds were abundant, they contributed 68.5 ± 22.7% of the diet while herbs accounted for 18.9 ± 12.9% on a DM. Although young bark and twigs were available in all seasons, the macaques only fed on them in winter. We present comparative data from the literature on the diets of M. mulatta and other Macaca species, to interpret this pattern of resource use within the framework of multilevel niche theory.

Sympatric speciation of spiny mice, Acomys, unfolded transcriptomically at Evolution Canyon, Israel.

Spiny mice, Acomys cahirinus, colonized Israel 30,000 y ago from dry tropical Africa and inhabited rocky habitats across Israel. Earlier, we had shown by mtDNA that A. cahirinus incipiently sympatrically speciates at Evolution Canyon I (EC I) in Mount Carmel, Israel because of microclimatic interslope divergence. The EC I microsite consists of a dry and hot savannoid "African" slope (AS) and an abutting humid and cool-forested "European" slope (ES). Here, we substantiate incipient SS in A. cahirinus at EC I based on the entire transcriptome, showing that multiple slope-specific adaptive complexes across the transcriptome result in two divergent clusters. Tajima's D distribution of the abutting Acomys interslope populations shows that the ES population is under stronger positive selection, whereas the AS population is under balancing selection, harboring higher genetic polymorphisms. Considerable sites of the two populations were differentiated with a coefficient of FST = 0.25-0.75. Remarkably, 24 and 37 putatively adaptively selected genes were detected in the AS and ES populations, respectively. The AS genes involved DNA repair, growth arrest, neural cell differentiation, and heat-shock proteins adapting to the local AS stresses of high solar radiation, drought, and high temperature. In contrast, the ES genes involved high ATP associated with energetics stress. The sharp ecological interslope divergence led to strong slope-specific selection overruling the interslope gene flow. Earlier tests suggested slope-specific mate choice. Habitat interslope-adaptive selection across the transcriptome and mate choice substantiate sympatric speciation (SS), suggesting its prevalence at EC I and commonality in nature.