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Pharmacology Research & Perspectives Jun 2022Compounds that induce 5-aminolevulinic acid [ALA] synthase-1 and/or cytochromes P-450 may induce acute porphyric attacks in patients with the acute hepatic porphyrias...
Compounds that induce 5-aminolevulinic acid [ALA] synthase-1 and/or cytochromes P-450 may induce acute porphyric attacks in patients with the acute hepatic porphyrias [AHPs]. Currently, there is no simple, robust model used to assess and predict the porphyrogenicity of drugs and chemicals. Our aim was to develop a fluorescence-based in vitro assay for this purpose. We studied four different hepatic cell culture models: HepG2 cells, LMH cells, 3D HepG2 organoids, and 3D organoids of primary liver cells from people without known disease [normal human controls]. We took advantage of the fluorescent properties of protoporphyrin IX [PP], the last intermediate of the heme biosynthesis pathway, performing fluorescence spectrometry to measure the intensity of fluorescence emitted by these cells treated with selected compounds of importance to patients with AHPs. Among the four cell culture models, the LMH cells produced the highest fluorescence readings, suggesting that these cells retain more robust heme biosynthesis enzymes or that the other cell models may have lost their inducibility of ALA synthase-1 [ALAS-1]. Allyl isopropyl acetamide [AIA], a known potent porphyrogen and inducer of ALAS-1, was used as a positive control to help predict porphyrogenicity for tested compounds. Among the tested compounds (acetaminophen, acetylsalicylic acid, β-estradiol, hydroxychloroquine sulfate, alpha-methyldopa, D (-) norgestrel, phenobarbital, phenytoin, sulfamethoxazole, sulfisoxazole, sodium valproate, and valsartan), concentrations greater than 0.314 mM for norgestrel, phenobarbital, phenytoin, and sodium valproate produced fluorescence readings higher than the reading produced by the positive AIA control. Porphyrin accumulation was also measured by HPLC to confirm the validity of the assay. We conclude that LMH cell cultures in multi-well plates are an inexpensive, robust, and simple system to predict the porphyrogenicity of existing or novel compounds that may exacerbate the AHPs.
Topics: Cell Culture Techniques; Heme; Hepatocytes; Humans; Liver; Norgestrel; Phenobarbital; Phenytoin; Porphobilinogen Synthase; Porphyrias, Hepatic; Valproic Acid
PubMed: 35445802
DOI: 10.1002/prp2.951 -
Journal of Dairy Science Apr 2020Klebsiella spp. are important opportunistic pathogens commonly defined as environmental clinical mastitis agents. Despite Klebsiella mastitis being clinically impairing...
Klebsiella spp. are important opportunistic pathogens commonly defined as environmental clinical mastitis agents. Despite Klebsiella mastitis being clinically impairing in cows and costly to the industry, only a few studies describe Klebsiella isolated from mastitis cases. The aim of this work was to characterize species of Klebsiella involved in clinical mastitis cases in Canada. Klebsiella isolated from clinical mastitis cases (n = 53) were identified to the species level using a biochemical test panel and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The rpoB gene sequence was used as the gold standard method and identified Klebsiella pneumoniae (n = 40), Klebsiella oxytoca (n = 9), Raoultella ornithinolytica (n = 2), and Raoultella planticola (n = 2). Raoultella, a genus closely related to Klebsiella, was also accurately identified using mass spectrometry but not via biochemical testing. Using the disc diffusion technique, 31 (58%) isolates were found to be susceptible to all antimicrobials tested (n = 18). The remaining 22 (42%) isolates were resistant to 1 or more of the following antimicrobials: kanamycin (2%), streptomycin (38%), spectinomycin (13%), sulfisoxazole (13%), and tetracycline (19%). The following antimicrobial resistance genes were identified: tetA, tetB, sul1, strA/strB, and aadA. Random amplified polymorphic DNA revealed the majority of our isolates as unrelated and having different patterns, indicating environmental contamination as the primary source of infection. All isolates were shown to be biofilm producers. In conclusion, although antimicrobial resistance was low for both Klebsiella and Raoultella species, genetically related Klebsiella spp. isolates appeared to be more resistant.
Topics: Animals; Anti-Bacterial Agents; Canada; Cattle; Drug Resistance, Bacterial; Female; Klebsiella; Mastitis, Bovine; Microbial Sensitivity Tests
PubMed: 32089315
DOI: 10.3168/jds.2019-17324 -
ACS Omega Dec 2023The search for novel drug scaffolds that can improve effectiveness and safety through drug conjugates is a promising approach. Consequently, drug conjugates constitute a...
Exploring the Potential of New Benzamide-Acetamide Pharmacophore Containing Sulfonamide as Urease Inhibitors: Structure-Activity Relationship, Kinetics Mechanism, and In Silico Studies.
The search for novel drug scaffolds that can improve effectiveness and safety through drug conjugates is a promising approach. Consequently, drug conjugates constitute a dynamic field of study and advancement within medicinal chemistry. This research demonstrates the conjugation of diclofenac and mefenamic acid with sulfa drugs and their screening for urease inhibition. These conjugates' structural confirmation was performed using elemental analysis and spectroscopic methods, including IR, H NMR, and C NMR. Diclofenac conjugated with sulfanilamide (4), sulfacetamide (10), and mefenamic acid conjugated with sulfanilamide (12), and sulfamethoxazole (17) was found potent and demonstrated urease inhibition competitively, with IC (μM) values 3.59 ± 0.07, 5.49 ± 0.34, 7.92 ± 0.27, and 8.35 ± 0.26, respectively. Diclofenac conjugated with sulfathiazole (6), sulfamerazine (8), and sulfaguanidine (11), while mefenamic acid conjugated with sulfisoxazole (13), sulfathiazole (14), and sulfadiazine (15) exhibited a mixed mode of urease inhibition. The IC (μM) values were 16.19 ± 0.21, 9.50 ± 0.28, 4.35 ± 0.23, 15.86 ± 0.25, 14.80 ± 0.27, and 7.92 ± 0.27, respectively. Furthermore, molecular docking studies were employed to predict the binding pose of competitive inhibitors at the urease active site. These conjugates generated stable complexes with the urease protein observed through molecular dynamics (MD) simulations, where no conformational changes occurred throughout the simulations. These results highlight the potential for approved therapeutic molecule conjugates to give rise to new categories of pharmacological agents for urease inhibition. The structural similarity of sulfonamides with urea allows them to compete with urea for binding to the active site of the urease enzyme. Sulfonamides and nonsteroidal anti-inflammatory drugs (NSAIDs) can interact hydrophobically with the active site of the urease enzyme, which may disturb its structure and catalytic activity. Therefore, these conjugates may be helpful in the development of novel pharmacological agents for the treatment of a variety of illnesses in which the urease enzyme is involved.
PubMed: 38075833
DOI: 10.1021/acsomega.3c07275 -
Veterinaria Italiana Dec 2021Salmonellosis is currently the second most common zoonosis in European Union but in the 6-years periods, between 2012 and 2017, there has been a significant decrease...
Salmonellosis is currently the second most common zoonosis in European Union but in the 6-years periods, between 2012 and 2017, there has been a significant decrease trend in the yearly number of infections caused by Salmonella. In Italy, S. Typhimurium and monophasic S. Typhimurium represent the most prevalent serotypes. In this paper, we investigated these two serovars isolated from 2012 to 2017 in Abruzzo and Molise regions. A set of 345 strains isolated from human sporadic cases, surface water, food and animals were collected and analyzed. Monophasic S. Typhimurium strains were found to be resistant to streptomycin, sulfisoxazole, ampicillin, tetracycline and nalidixic acid, while S. Typhimurium isolates showed high levels of resistance to tetracycline, sulfisoxazole and ampicillin. The 5-loci Multilocus Variable-Number Tandem Repeat Analysis (MLVA) identified 88 genotypes (GT), six of which were common for the two serovars. Several MLVA profiles were shared by human and non-human isolates. MLVA had sufficient typing resolution for epidemiological studies of S. Typhimurium but demonstrated poor discriminatory in trace-back study of monophasic S. Typhimurium.
Topics: Ampicillin; Animals; Anti-Bacterial Agents; Italy; Salmonella typhimurium; Sulfisoxazole; Tetracyclines
PubMed: 35593494
DOI: 10.12834/VetIt.2060.12083.1 -
Biology Jul 2021Clinical management of cancer-associated cachexia, a multi-organ wasting syndrome, has been challenging without effective treatment strategies. An effective treatment...
Clinical management of cancer-associated cachexia, a multi-organ wasting syndrome, has been challenging without effective treatment strategies. An effective treatment that directly targets cancer-induced wasting is desperately needed to improve the quality of life and the survival of cancer patients. Recently, an antibiotic SFX was shown to have anti-tumour and anti-metastatic effects in mouse models of breast cancer. Hence, in this study, we examined the efficacy of SFX in the treatment of cancer-induced cachexia. C26 cachexic mice models were administered with SFX, and the tumour volume and body weight were regularly measured. Blood glucose, skeletal muscles, and adipose tissue were examined at the endpoint. Contrary to a previous study, SFX did not reduce the tumour volume in mice bearing C26 cells. Administration of SFX neither revealed any survival benefit nor rescued C26 cachectic mice from muscle wasting. Interestingly, SFX administration partially rescued (~10%) tumour-induced weight loss by preserving both the subcutaneous and intestinal fat mass. Together, these results suggest that the administration of SFX could partially rescue cancer-induced weight loss by inhibiting lipolysis. As anti-cachexia therapies are scarce, the results could facilitate the design of combinatorial therapies involving SFX, standard-of-care chemotherapeutics, and drugs that inhibit muscle atrophy for the treatment of cancer cachexia.
PubMed: 34439933
DOI: 10.3390/biology10080700 -
Frontiers in Microbiology 2018This study aimed to investigate the prevalence, serotype distribution, and antibiotic resistance, and to characterize the extended spectrum β-lactamases (ESBLs)...
This study aimed to investigate the prevalence, serotype distribution, and antibiotic resistance, and to characterize the extended spectrum β-lactamases (ESBLs) producing isolates from chicken and pork meats from retail markets in Guangdong province, China. A total of 903 retail meat samples (475 chicken and 428 pork meats) were obtained from six cities (Guangzhou, Shenzhen, Heyuan, Shaoguan, Foshan, and Yunfu) of Guangdong province between May 2016 and April 2017. High levels of contamination were detected in chicken (302/475, 63.6%) and pork (313/428, 73.1%). Thirty-eight serotypes were identified in 615 detected , and the serotypes varied greatly between chicken and pork samples. Agona (55/302, 18.2%), Corvallis (45/302, 14.9%), Kentucky (38/302, 12.6%), Mbandaka (32/302, 10.6%) was the dominant serotypes in chicken samples. However, Typhimurium (78/313, 24.9%), Rissen (67/313, 24.1%), Derby (66/313, 21.1%), and London (48, 15.3%) were the most common in pork samples. High rates of antibiotic resistance were found to sulfisoxazole (468/615, 76.1%), tetracycline (463/615, 75.3%), ampicillin (295/615, 48.0%), and ofloxacin (275/615, 44.7%). Notably, antimicrobial susceptibility tests identified resistance to polymyxin B (12/615, 2.0%) and imipenem (3/615, 0.5%). Multidrug-resistance (MDR) was detected in isolated from chicken (245/302, 81.1%) and pork (229/313, 73.2%). The resistance rate of different serotypes varied widely. Especially, isolates such as Typhimurium, Agona, orvallis and Kentucky exhibited highly resistance to antibiotics. The MDR rate of isolates from chicken was significantly higher than that from pork isolates ( < 0.05). Twenty-one isolates were identified as ESBLs-producing, covering six serotypes and displaying different pulse field gel electrophoresis (PFGE) genotypes. was the dominant ESBLs gene (9/21, 42.9%), followed by (5/21, 23.8%). This study indicated that was widespread in chicken and pork from retail markets in Guangdong province and the isolates showed high multidrug-resistance, especially the known multidrug-resistant serotypes. Therefore, it is important to focus on serotypes and strengthen the long-term monitoring of MDR serotypes in animal-derived foods.
PubMed: 30258419
DOI: 10.3389/fmicb.2018.02104 -
Nature Communications Mar 2019Inhibitors of the secretion of cancer exosomes, which promote cancer progression and metastasis, may not only accelerate exosome biology research but also offer...
Inhibitors of the secretion of cancer exosomes, which promote cancer progression and metastasis, may not only accelerate exosome biology research but also offer therapeutic benefits for cancer patients. Here we identify sulfisoxazole (SFX) as an inhibitor of small extracellular vesicles (sEV) secretion from breast cancer cells through interference with endothelin receptor A (ETA). SFX, an FDA-approved oral antibiotic, showed significant anti-tumor and anti-metastatic effects in mouse models of breast cancer xenografts, the reduced expression of proteins involved in biogenesis and secretion of sEV, and triggered co-localization of multivesicular endosomes with lysosomes for degradation. We demonstrate the important role of ETA, as target of SFX, by gain- and loss-of-function studies of the ETA protein, through a direct binding assay, and pharmacological and genetic approaches. These findings may provide a foundation for sEV-targeted cancer therapies and the mechanistic studies on sEV biology.
Topics: Animals; Anti-Infective Agents; Breast Neoplasms; Cell Line, Tumor; Endosomes; Extracellular Vesicles; Female; Humans; Lysosomes; MCF-7 Cells; Male; Mice; Mice, Nude; Neoplasm Metastasis; Organelle Biogenesis; Receptor, Endothelin A; Sulfisoxazole; Xenograft Model Antitumor Assays
PubMed: 30918259
DOI: 10.1038/s41467-019-09387-4 -
MSphere 2016The effects of animal agriculture on the spread of antibiotic resistance (AR) are cross-cutting and thus require a multidisciplinary perspective. Here we use ecological,...
The effects of animal agriculture on the spread of antibiotic resistance (AR) are cross-cutting and thus require a multidisciplinary perspective. Here we use ecological, epidemiological, and ethnographic methods to examine populations of Escherichia coli circulating in the production poultry farming environment versus the domestic environment in rural Ecuador, where small-scale poultry production employing nontherapeutic antibiotics is increasingly common. We sampled 262 "production birds" (commercially raised broiler chickens and laying hens) and 455 "household birds" (raised for domestic use) and household and coop environmental samples from 17 villages between 2010 and 2013. We analyzed data on zones of inhibition from Kirby-Bauer tests, rather than established clinical breakpoints for AR, to distinguish between populations of organisms. We saw significantly higher levels of AR in bacteria from production versus household birds; resistance to either amoxicillin-clavulanate, cephalothin, cefotaxime, and gentamicin was found in 52.8% of production bird isolates and 16% of household ones. A strain jointly resistant to the 4 drugs was exclusive to a subset of isolates from production birds (7.6%) and coop surfaces (6.5%) and was associated with a particular purchase site. The prevalence of AR in production birds declined with bird age (P < 0.01 for all antibiotics tested except tetracycline, sulfisoxazole, and trimethoprim-sulfamethoxazole). Farming status did not impact AR in domestic environments at the household or village level. Our results suggest that AR associated with small-scale poultry farming is present in the immediate production environment and likely originates from sources outside the study area. These outside sources might be a better place to target control efforts than local management practices. IMPORTANCE In developing countries, small-scale poultry farming employing antibiotics as growth promoters is being advanced as an inexpensive source of protein and income. Here, we present the results of a large ecoepidemiological study examining patterns of antibiotic resistance (AR) in E. coli isolates from small-scale poultry production environments versus domestic environments in rural Ecuador, where such backyard poultry operations have become established over the past decade. Our previous research in the region suggests that introduction of AR bacteria through travel and commerce may be an important source of AR in villages of this region. This report extends the prior analysis by examining small-scale production chicken farming as a potential source of resistant strains. Our results suggest that AR strains associated with poultry production likely originate from sources outside the study area and that these outside sources might be a better place to target control efforts than local management practices.
PubMed: 27303705
DOI: 10.1128/mSphere.00021-15 -
Canadian Journal of Veterinary Research... Apr 2019The objectives of this study were to describe the frequency of antimicrobial resistance (AMR) in and spp. isolates in fecal samples from beef cow-calf herds and to...
The objectives of this study were to describe the frequency of antimicrobial resistance (AMR) in and spp. isolates in fecal samples from beef cow-calf herds and to examine the associations between herd management practices, reported antimicrobial use, and AMR. Baseline prevalence data are needed to evaluate the effectiveness of antimicrobial stewardship programs. A pooled fecal sample, representing 20 cows, was collected from each of 107 herds during pregnancy testing. In the 305 recovered isolates (maximum 3 per herd), resistance to ≥ 1 antimicrobial was identified in 12 isolates [4%, 95% confidence interval (CI): 2% to 7%] from 105 herds (11%, 95% CI: 7% to 19%). The most common resistances identified in isolates were to tetracycline (3%) and to both streptomycin and sulfisoxazole (3%). Only 1 isolate was resistant to an antimicrobial of very high importance to human health - amoxicillin/clavulanic acid. However, 2 isolates had intermediate susceptibility to ciprofloxacin. Resistance to 1 antimicrobial was identified in 16 of 87 spp. isolates (18%, 95% CI: 11% to 28%) from 87 herds. Resistance to tetracycline was reported in 15% of spp. isolates and to nalidixic acid in 3.4%. Herds in which cows were treated with florfenicol were more likely to have resistance to ≥ 2 antimicrobials (OR 7.1, 95% CI: 1.1 to 57, = 0.03). Herds with calf mortality of > 5% were more likely to have with resistance to streptomycin and sulfisoxazole [odds ratio (OR): 7.8, = 0.03]. The results of this study are consistent with previous reports from western Canada and provide a starting point for designing an ongoing antimicrobial surveillance program.
Topics: Alberta; Animals; Anti-Bacterial Agents; Cattle; Drug Resistance, Bacterial; Drug Utilization; Escherichia coli; Feces; Female; Manitoba; Microbial Sensitivity Tests; Saskatchewan
PubMed: 31097869
DOI: No ID Found -
Revue Scientifique Et Technique... Dec 2019Antibiotics used in animals may be found in food of animal origin and pose a risk to human health. The aim of this study was to screen for antibiotic residues in broiler...
Antibiotics used in animals may be found in food of animal origin and pose a risk to human health. The aim of this study was to screen for antibiotic residues in broiler chickens and milk. Two hundred and twenty-one samples were collected (71 samples of chicken breast meat, 117 samples of raw cow's milk and 33 samples of raw goat's milk). The chicken meat samples underwent a microbiological analysis, followed by a physical/chemical analysis using high-performance liquid chromatography (HPLC). The milk samples were screened using a commercial microbiological test, followed by a further test for residues of beta-lactams and tetracyclines. For chicken meat, 32.39% of the samples were positive, with 56.52% of these samples containing aminoglycosides, 52.17% containing sulphonamides, 30.43% containing beta-lactams and/or tetracyclines and 21.73% containing macrolides. The concentrations of amoxicillin, penicillin G, erythromycin and sulfisoxazole exceeded the maximum residue limits laid down in European regulations in 28.57%, 85.71%, 80% and 91.66% of samples, respectively. The results of the test on milk showed that 12.6% of samples were contaminated by inhibiting substances. Betalactams and tetracyclines were present in 26.32% and 15.79% of the samples analysed, respectively. These results indicate that the contamination of chicken meat and milk is due to non-compliance with administration procedures and inappropiate use of antibiotics.
Topics: Algeria; Animals; Anti-Bacterial Agents; Cattle; Chickens; Drug Residues; Female; Food Contamination; Humans; Meat; Milk
PubMed: 32286562
DOI: 10.20506/rst.38.3.3031