Role of Gut Microbiome in Neoadjuvant Chemotherapy Response in Urothelial Carcinoma: A Multi-Institutional Prospective Cohort Evaluation.

Neoadjuvant chemotherapy (NAC) is linked with clinical advantages in urothelial carcinoma for patients with muscle-invasive bladder cancer (MIBC). Despite comprehensive research into the influence of tumor mutation expression profiles and clinicopathological factors on chemotherapy response, the role of the gut microbiome (GM) in bladder cancer(BC) chemotherapy response remains poorly understood. This study examines the variance in the gut microbiome(GM) of BC patients compared to healthy adults, and investigates GM compositional differences between patients who respond to chemotherapy versus those who exhibit residual disease. Our study reveals distinct clustering, effectively separating the BC and healthy cohorts. However, no significant differences were observed between chemotherapy responders and non-responders within community subgroups. Machine Learning models based on responder status outperformed clinical variables in predicting complete response (AUC 0.88 vs AUC 0.50), although no single microbial species emerged as a fully reliable biomarker. The evaluation of short-chain fatty acid (SCFA) concentration in blood and stool revealed no correlation with responder status. Still, SCFA analysis showed a higher abundance of Akkermansia (rs = 0.51, p = 0.017) and Clostridia (rs = 0.52, p = 0.018), which correlated with increased levels of detectable fecal isobutyric acid. Higher levels of fecal Lactobacillus (rs = 0.49, p=0.02) and Enterobacteriaceae (rs = 0.52, p < 0.03) correlated with increased fecal propionic acid. In conclusion, our study constitutes the first large-scale, multi-center assessment of GM composition, suggesting the potential for a complex microbial signature to predict patients more likely to respond to NAC based on multiple taxa.

Tri-state logic computation by activating DNA origami chains.

The invention of DNA nanotechnology has enabled molecular computation as a promising substitute for traditional semiconductors which are limited to two-dimensional architectures and by heating problems resulting from densification. Current studies of logic gates achieved using DNA molecules are predominately focused on two-state operations (AND, OR, etc.); however, realizing tri-state logic (high impedance Z) in DNA computation is understudied. Here we actively fold DNA origami chain-like hinged rods to induce conformational changes that return tri-state logic signals. We use rigid six helix-bundle (6HB) DNA origami to self-assemble a linear trimer chain as a circuit platform with functional single-stranded (ss) DNA near each semi-flexible hinge. The presence or absence of ssDNA enable and input strands allows hybridization to take place at the hinges, activating one fold (0) or two folds (1) from the straight linear geometry (defined as High-Z) of the trimer chain. We design two different tri-state logic gate platforms, buffer and inverter, with corresponding enable/input ssDNA to unambiguously return tri-state signals, characterized by Atomic Force Microscopy (AFM) and/or agarose gel electrophoresis (GEL). Our work on tri-state logic significantly enhances DNA computation beyond the current two-state Boolean logic with both research and industrial applications, including cellular treatments and living matter utilizing the biocompatibility of DNA molecules.

The Response of the Urinary Microbiome to Botox.

INTRODUCTION AND HYPOTHESIS: Our objective was to evaluate if botox alters the urinary microbiome of patients with overactive bladder and whether this alteration is predictive of treatment response. METHODS: This multicenter prospective cohort study included 18-89-year-old patients undergoing treatment for overactive bladder with 100 units of botox. Urine samples were collected by straight catheterization on the day of the procedure (S1) and again 4 weeks later (S2). Participants completed the Patient Global Impression of Improvement form at their second visit for dichotomization into responders and nonresponders. The microbiome was sequenced using 16s rRNA sequencing. Wilcoxon signed rank and Wilcoxon rank sum were used to compare the microbiome, whereas chi-square, Wilcoxon rank sum, and the independent t-test were utilized for clinical data. RESULTS: Sixty-eight participants were included in the analysis. The mean relative abundance and prevalence of Beauveria bassiana, Xerocomus chrysenteron, Crinipellis zonata, and Micrococcus luteus were all found to increase between S1 and S2 in responders; whereas in nonresponders the mean relative abundance and prevalence of Pseudomonas fragi were found to decrease. The MRA and prevalence of Weissella cibaria, Acinetobacter johnsonii, and Acinetobacter schindleri were found to be greater in responders than nonresponders at the time of S1. Significant UM differences in the S1 of patients who did (n = 5) and did not go on to develop a post-treatment UTI were noted. CONCLUSIONS: Longitudinal urobiome differences may exist between patients who do and do not respond to botox.

Antifungal Activity of Efinaconazole Compared to Fluconazole, Itraconazole, and Terbinafine against Terbinafine- and Itraconazole-Resistant and -Susceptible Clinical Isolates of Dermatophytes, Candida, and Mold.

BACKGROUND: Recently, an increasing number of resistant-to-terbinafine dermatophytosis cases have been reported. Thus, identifying an alternative antifungal agent that possesses a broad-spectrum activity, including against resistant strains, is needed. METHODS: In this study, we compared the antifungal activity of efinaconazole to fluconazole, itraconazole, and terbinafine against clinical isolates of dermatophyte, Candida, and molds using in vitro assays. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of each antifungal was quantified and compared. Both susceptible and resistant clinical isolates of Trichophyton mentagrophytes (n=16), T. rubrum (n=43), T. tonsurans (n=18), T. violaceum (n=4), Candida albicans (n=55), C. auris (n=30), Fusarium sp., Scedosporium sp., and Scopulariopsis sp. (n=15 for each) were tested. RESULTS: Our data shows that efinaconazole was the most active antifungal, compared to the other agents tested, against dermatophytes with MIC50 and MIC90 (Concentration that inhibited 50% and 90% of strains tested, respectively) values of 0.002 and 0.03 µg/ml, respectively. Fluconazole, itraconazole and terbinafine showed MIC50 and MIC90 values of 1 and 8 µg/ml, 0.03 and 0.25 µg/ml, and 0.031 and 16 µg/ml, respectively. Against Candida isolates, efinaconazole MIC50 and MIC90 values were 0.016 and 0.25 µg/ml, respectively, whereas fluconazole, itraconazole and terbinafine had MIC50 and the MIC90 values of 1 and 16 µg/ml, 0.25 and 0.5 µg/ml, and 2 and 8 µg/ml, respectively. Against various mold species, efinaconazole MIC values ranged from 0.016 and 2 µg/ml, compared to 0.5 to greater than 64 µg/ml for the comparators. CONCLUSIONS: efinaconazole showed superior potent activity against a broad panel of susceptible and resistant dermatophyte, Candida, and mold isolates.

Role of Gut Microbiome in Neoadjuvant Chemotherapy Response in Urothelial Carcinoma: A Multi-Institutional Prospective Cohort Evaluation.

Treatment with neoadjuvant chemotherapy (NAC) in muscle invasive bladder cancer (MIBC) is associated with clinical benefit in urothelial carcinoma. While extensive research evaluating role of tumor mutational expression profiles and clinicopathologic factors into chemoresponse has been published, the role of gut microbiome (GM) in bladder cancer in chemoresponse has not been thoroughly evaluated. A working knowledge of the microbiome and its effect on all forms of cancer therapy in BC is critical. Here we examine gut microbiome of bladder cancer patients undergoing NAC. Overall, there was no significant difference in alpha and beta diversity by responder status. However, analysis of fecal microbiome samples showed that a higher abundance of Bacteroides within both institutional cohorts during NAC was associated with residual disease at the time of radical cystectomy regardless of chemotherapy regimen. Group community analysis revealed presence of favorable microbial subtypes in complete responders. Finally, fecal microbial composition outperformed clinical variables in prediction of complete response (AUC 0.88 vs AUC 0.50), however, no single microbial species could be regarded as a fully consistent biomarker. Microbiome-based community signature as compared to single microbial species is more likely to be associated as the link between bacterial composition and NAC response.

Carbapenem Resistance in Gram-Negative Bacteria: A Hospital-Based Study in Egypt.

Background and Objectives: The global spread of carbapenem resistance and the resulting increase in mortality forced the World Health Organization (WHO) to claim carbapenem-resistant enterobacteriaceae (CRE) as global priority pathogens. Our study aimed to determine the prevalence of carbapenemase-encoding genes and major plasmid incompatibility groups among Gram-negative hospital-based isolates in Egypt. Material and Methods: This cross-sectional study was carried out at Mansoura University Hospitals over 12 months, from January to December 2019. All the isolates were tested for carbapenem resistance. The selected isolates were screened by conventional polymerase chain reaction (PCR) for the presence of carbapenemase genes, namely blaKPC, blaIMP, blaVIM, and blaNDM-1. PCR-based plasmid replicon typing was performed using the commercial PBRT kit. Results: Out of 150 isolates, only 30 (20.0%) demonstrated carbapenem resistance. Klebsiella pneumoniae was the most resistant of all isolated bacteria, and blaNDM was the predominant carbapenemases gene, while the most prevalent plasmid replicons were the F replicon combination (FIA, FIB, and FII) and A/C. Plasmids were detected only in Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, and Pseudomonas aeruginosa. Remarkably, we found a statistically significant association between carbapenemase genes and plasmid replicons, including blaNDM, IncA/C, and IncX. Conclusions: Our study demonstrated an alarming rise of plasmid-mediated carbapenem-resistant bacteria in our locality. The coexistence of resistance genes and plasmids highlights the importance of a targeted antibiotic surveillance program and the development of alternative therapeutic options at the local and international levels. Based on our results, we suggest a large-scale study with more Enterobacteriaceae isolates, testing other carbapenemase-encoding genes, and comparing the replicon typing method with other plasmid detection methods. We also recommend a national action plan to control the irrational use of antibiotics in Egypt.

Ziziphus spina-christi extract-stabilized novel silver nanoparticle synthesis for combating Fusarium oxysporum-causing pepper wilt disease: in vitro and in vivo studies.

The novelty of the present study is studying the ability of aqueous Ziziphus spina-christi leaves' extract (ZSCE) to produce eco-friendly and cost-effective silver nanoparticles (Ag NPs) against Fusarium wilt disease. Phytochemical screening of ZSCE by HPLC showed that they contain important antimicrobial substances such as Rutin, Naringin, Myricetin, Quercetin, Kaempferol, Hesperidin, Syringeic, Eugenol, Pyrogallol, Gallic and Ferulic. Characterization methods reveal a stable Ag NPs with a crystalline structure, spherical in shape with average particle size about 11.25 nm. ZSCE and Ag NPs showed antifungal potential against F. oxysporum at different concentrations with MIC of Ag NPs as 0.125 mM. Ag NPs treatment was the most effective, as it gave the least disease severity (20.8%) and the highest protection rate (75%). The application of ZSCE or Ag NPs showed a clear recovery, and its effectiveness was not limited for improving growth and metabolic characteristics only, but also inducing substances responsible for defense against pathogens and activating plant immunity (such as increasing phenols and strong expression of peroxidase and polyphenol oxidase as well as isozymes). Owing to beneficial properties such as antifungal activity, and the eco-friendly approach of cost and safety, they can be applied in agricultural field as novel therapeutic nutrients.

Efficacy of chlorhexidine in advanced performance technology formulation in decolonizing the skin using Candida auris skin colonization mouse model.

The incidence of Candida auris, an emerging multidrug resistant fungal species, is increasing. The ability of this yeast to colonize the human skin could lead to infections. Identifying agents to reduce the skin fungal burden is critical. Chlorhexidine formulated in a new Advanced Performance Technology formulation (APT-CH) was significantly more effective than untreated controls. Additional studies are warranted.

Analysis of Gut Microbiome Using Fecal Samples.

With the gut microbiome being implicated in many diseases and even offering the potential for future therapies, its study has been gaining interest over the years. However, to gain more in-depth insight and understanding of the impact of the microbiome on our health, it is critical to have accurate microbial data that is reproducible and comparable across studies. Variation in experimental techniques, particularly DNA extraction and sequencing methods, may result in variability and the inability to compare results. In this chapter, we describe in detail our microbiome analysis methodology focusing on stool samples.

The Mycobiome: Cancer Pathogenesis, Diagnosis, and Therapy.

Cancer is among the leading causes of death globally. Despite advances in cancer research, a full understanding of the exact cause has not been established. Recent data have shown that the microbiome has an important relationship with cancer on various levels, including cancer pathogenesis, diagnosis and prognosis, and treatment. Since most studies have focused only on the role of bacteria in this process, in this article we review the role of fungi-another important group of the microbiome, the totality of which is referred to as the "mycobiome"-in the development of cancer and how it can impact responses to anticancer medications. Furthermore, we provide recent evidence that shows how the different microbial communities interact and affect each other at gastrointestinal and non-gastrointestinal sites, including the skin, thereby emphasizing the importance of investigating the microbiome beyond bacteria.

Adherence and Effectiveness of HBV Vaccination among Healthcare Workers in Egypt.

CONTEXT: Healthcare workers (HCWs) face a potential risk of acquiring different bloodborne pathogens, by occupational exposure to contaminated blood and body fluids. Hepatitis B vaccine is a safe, effective method of conferring long-term protection against HBV infection. AIMS: The study aimed to assess the adherence and effectiveness of HBV vaccination among HCWs at Gastrointestinal Surgical Center, Mansoura University, Egypt. SETTINGS AND DESIGN: A prospective descriptive study was carried out between June 2019 and December 2019 at Gastrointestinal Surgical Center, Mansoura University, Egypt. METHODS AND MATERIAL: All HCWs with anti-HBs levels below 10 mIU/mL were advised to receive 3 doses of recombinant HBV vaccine, at 0, 1, and 6 months. The anti-HBs levels were checked 3 months after the third dose of the HBV vaccine. STATISTICAL ANALYSIS USED: Data was analyzed using the Statistical Package of Social Science (SPSS) program for windows (version 16). RESULTS: A total of 442 healthcare providers were included. Most of them completed the 3 doses of the vaccine (81.7%), 10.2% refused the vaccine, while 0.9% and 7.2% received 1 and 2 doses, respectively. Odds of vaccination were the highest (88.1%, 273/310) among nurses (OR, 4.7; 95% CI, 2.6-5.2; P ≤ 0.001). The overall anti-HBs positivity of 97% (350/361) was observed. The main reasons for not being vaccinated included the fear of vaccine side effects 25/81 (30.9%) and lack of trust in the vaccine effectiveness 18/81 (22.2%). CONCLUSIONS: The outcome of the present study emphasizes the need to apply alternative and innovative measures to build a positive attitude toward the HBV vaccine among HCWs.

Synthesis and Antiproliferative Evaluation of 2-Deoxy-N-glycosylbenzotriazoles/imidazoles.

A series of 2-deoxy-2-iodo-α-d-mannopyranosylbenzotriazoles was synthesized using the benzyl, 4,6-benzylidene and acetyl protected D-glucal in the presence of N-iodosuccinimide (NIS). Subsequent removal of the iodine at the C-2 position using tributyltin hydride under free radical conditions afforded the 2-deoxy-α-d-glucopyranosylbenzotriazoles in moderate to high yields. This method was extended to the preparation of substituted 2-deoxy-ß-d-glucopyranosylimidazoles as well. The stereoselectivity of the addition reaction and the effect of the protecting group and temperature on anomer distribution of the benzotriazole series were also investigated. The anticancer properties of the newly synthesized compounds were evaluated in a series of viability studies using HeLa (human cervical adenocarcinoma), human breast and lung cancer cell lines. The N-[3,4,6-tri-O-benzyl-2-deoxy-α-d-glucopyranosyl]-1H-benzotriazole and the N-[3,4,6-tri-O-acetyl-2-deoxy-α-d-glucopyranosyl]-2H-benzotriazole were found to be the most potent cancer cell inhibitors at 20 µM concentrations across all four cell lines.

Mechanistic insights into acetaminophen removal on cashew nut shell biomass-derived activated carbons.

Activated carbons prepared from cashew nut shells by chemical activation with phosphoric acid were tested for the removal of acetaminophen. It was found that an increase in carbonization temperature resulted in increased pore volume and decreased amount of surface functional groups. Potentiometric titration analysis indicated that the majority of surface groups on carbons are acidic. Detailed surface characterization by FT-IR, XPS, and thermal analyses indicated the involvement of surface functional groups in the removal of acetaminophen either via hydrogen bonding or by acid hydrolysis. The carbon obtained at 600 °C, which contains high amount of carboxylic groups and high pore volume, exhibited the highest adsorption capacity. For this carbon, the removal of acetaminophen took place mostly via acid hydrolysis with the formation of p-aminophenol and acetic acid adsorbed on the surface. Carbon obtained at 400 °C was found to have the highest density of acidic functional groups, which resulted in dimerization reactions and pore blockage. No direct correlation was observed between the adsorption capacities of carbons and their textural or surface characteristics. This suggests the complexity of acetaminophen removal by the cashew nut shell-derived activated carbons, governed by their surface chemistry and supported by high surface area accessible via micro/mesopores.