Despite the natural ability of the immune system to recognize cancer and, in some patients, even to eliminate it, cancer cells have acquired numerous evading mechanisms. With the increasing knowledge and focus shifting from targeting rapidly proliferating cells with chemotherapy to modulating the immune system, there have been recent efforts to integrate (e.g., simultaneously or sequentially) various therapeutic approaches. Combining the oncolytic activity of some chemotherapeutics with immunostimulatory molecules, so-called chemoimmunotherapy, is an attractive strategy. An example of such an immunostimulatory molecule is polyinosinic:polycytidylic acid [Poly(I:C)], a synthetic analogue of double-stranded RNA characterized by rapid nuclease degradation hampering its biological activity. This study investigated the possible interactions of tetracycline and anthracycline chemotherapeutics with different commercial Poly(I:C) molecules and protection against nuclease degradation. Fluorescence spectroscopy and circular dichroism revealed an interaction of all of the selected chemotherapeutics with Poly(I:C)s and the ability of doxycycline and minocycline to prolong the resistance to RNase cleavage, respectively. The partial protection was observed in vitro as well.

• Publication type
• Journal Article MeSH

OBJECTIVES: The aim of this study is to determine the incidence of Mycoplasma pneumoniae pneumonia (MPP) in children and adolescents in Prague, Czech Republic, between January and July 2024, and to compare the findings with data from the preceding period. METHODS: A retrospective analysis of data of paediatric patients at our single tertiary care facility was conducted. Two distinct patient cohorts were subjected to analysis: the first comprising individuals who had been hospitalised between January 2019 and July 2024, and the second consisting of outpatients who had been treated during the periods of January to July 2023 and January to July 2024. RESULTS: A 12.3-fold increase in the number of outpatients diagnosed with MPP was observed between January and July 2024 in comparison to the same period in 2023, with 111 cases reported in 2024 versus 9 cases in 2023. A total of 23 patients were hospitalised with MPP between January 2019 and July 2024, with 15 of these hospitalisations having occurred between January and July 2024. The median age was 12 years, with an age range of 1 to 17 years. The majority of cases presented with a high fever, chest pain, and required oxygen support. A failure of the clarithromycin treatment was observed, resulting in 19.48% of doxycycline prescriptions being issued due to a prior failure of clarithromycin treatment. During the monitoring period, no cases of treatment failure with doxycycline were documented. CONCLUSION: The present study demonstrates an emerging trend of increased incidence of Mycoplasma pneumoniae pneumonia in the paediatric population during the initial seven months of 2024 in the Czech Republic. Doxycycline has been demonstrated to be the optimal antibiotic for the treatment of MPP and in accordance with the prevailing practice in other states it should be included in the therapeutic regimen even in children under the age of eight. The authors put forward recommendations for the implementation of measures aimed at reducing the negative impact of MPP on public health.

• Keywords
• Mycoplasma pneumoniae, Mycoplasma pneumoniae pneumonia, atypical pneumonia, children, doxycycline, incidence, paediatric population,
• MeSH
• Anti-Bacterial Agents * therapeutic use   MeSH
• Tertiary Care Centers MeSH
• Child MeSH
• Hospitalization statistics & numerical data   MeSH
• Incidence MeSH
• Infant MeSH
• Humans MeSH
• Adolescent MeSH
• Mycoplasma pneumoniae MeSH
• Pneumonia, Mycoplasma * epidemiology   drug therapy   MeSH
• Child, Preschool MeSH
• Retrospective Studies MeSH
• Check Tag
• Child MeSH
• Infant MeSH
• Humans MeSH
• Adolescent MeSH
• Male MeSH
• Child, Preschool MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Czech Republic epidemiology   MeSH
• Names of Substances
• Anti-Bacterial Agents * MeSH

In this transdisciplinary study, we investigated, using genomic tools and physico-chemical parameters, the effect of Moringa oleifera seed (MOS) on the removal of microorganisms and pharmaceutic residues (antibiotics), and also the development of antibiotic-resistant genes (ARGs) in water samples from a domestic wastewater treatment plant (WWTP) prototype. Water samples were analyzed with and without the addition of powder of MOS. The results showed that MOS addition reduced the total bacterial load from 1.73 × 1010 ± 3.21 × 109 to 6.67 × 106 ± 5.77 × 106 CFU/L, while fecal coliforms and Escherichia coli were removed with efficiencies of 99% and 57%, respectively. Furthermore, MOS treatment resulted in a reduction in fecal coliforms and E. coli resistant to ampicillin by about 100% and 96%, respectively. The results indicated that ciprofloxacin removal efficiencies at 29 °C were over 93% (fecal coliforms) and 68% (E. coli) with doxycycline. Adding MOS significantly reduced the copy number of the 16S rRNA gene and the genes conferring resistance to β-lactam (blaCTX-M, blaSHV, and blaTEM). However, MOS does not reveal real effectiveness on removal of pollutants (phosphorus and nitrates) contrary to what was expected. Additional studies are needed for confirmation from our observations. The findings of this study, whatever the functioning conditions (not optimal) of the prototype followed over 4 years, confirmed that MOS is potentially an effective natural and environmentally friendly coagulant that could be applied to wastewater treatment in low-income countries to remove or minimize multiple pollutants and control ARG spread. To promote sustainable development, this small-scale study provides guidance for designing infrastructure in resource-limited locations to take advantage of MOS effects in wastewater treatments.

• Keywords
• Moringa oleifera seeds, Antibiotic resistant, Genomic, Natural coagulant, Pharmaceutic residues, Pollutants, Prototype, Resistant genes, Wastewater,
• MeSH
• Anti-Bacterial Agents pharmacology   MeSH
• Water Purification methods   MeSH
• Escherichia coli * MeSH
• Moringa oleifera * MeSH
• Waste Disposal, Fluid methods   MeSH
• Wastewater * chemistry   MeSH
• Seeds * MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Wastewater * MeSH

OBJECTIVES: To evaluate the efficacy and tolerability of a single dose of oral cefixime 800 mg plus oral doxycycline 100 mg twice a day for 7 days, compared with a recommended single dose of ceftriaxone plus single dose of oral azithromycin, for treatment of uncomplicated urogenital, rectal, or pharyngeal gonorrhoea. METHODS: A noninferiority, open-label, multicentre randomized controlled trial was conducted in Prague, Czech Republic. Some 161 patients, 18-65 years of age diagnosed with uncomplicated urogenital, rectal, or pharyngeal gonorrhoea by nucleic acid amplification test (NAAT) were randomized to treatment with single dose of cefixime 800 mg plus doxycycline 100 mg twice a day for 1 week or a single dose of ceftriaxone 1 g intramuscularly plus single dose of azithromycin 2 g. The primary outcome was the number of participants with negative culture and NAAT at 1 week and 3 weeks, respectively, after treatment initiation. RESULTS: In all, 161 patients were randomized and 152 were included in per-protocol analyses. All 76 (100%; 95% CI, 0.95-1.00) patients treated with ceftriaxone plus azithromycin achieved negative cultures and NAAT after treatment. In the cefixime plus doxycycline arm at week 1, culture was negative in all 76 (100%) patients; at week 3, culture was negative in 70 of the 76 patients (92%; 95% CI, 0.84-0.97) and NAAT negative in 66 of the 76 patients (87%; 95% CI, 0.77-0.94). At week 3, culture and NAAT were negative in 65 of the 76 patients (86%; 95% CI, 0.76-0.93). Per-protocol risk difference was 14.5%; 95% CI, 6.56-22.38. All treatment failures observed in the cefixime arm were pharyngeal gonorrhoea cases. DISCUSSION: The combination of cefixime and doxycycline did not achieve noninferiority to ceftriaxone and azithromycin for treatment of gonorrhoea when including pharyngeal gonorrhoea. It did, however, show high efficacy for urogenital and rectal gonorrhoea.

• Keywords
• Azithromycin, Cefixime, Ceftriaxone, Doxycycline, Neisseria gonorrhoeae, Treatment,
• MeSH
• Anti-Bacterial Agents therapeutic use   MeSH
• Azithromycin therapeutic use   MeSH
• Cefixime therapeutic use   MeSH
• Ceftriaxone * MeSH
• Adult MeSH
• Doxycycline therapeutic use   MeSH
• Gonorrhea * drug therapy   microbiology   MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Young Adult MeSH
• Neisseria gonorrhoeae MeSH
• Aged MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Young Adult MeSH
• Aged MeSH
• Publication type
• Journal Article MeSH
• Equivalence Trial MeSH
• Multicenter Study MeSH
• Randomized Controlled Trial MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Azithromycin MeSH
• Cefixime MeSH
• Ceftriaxone * MeSH
• Doxycycline MeSH

Mitochondria are central for cancer responses to therapy-induced stress signals. Refractory tumors often show attenuated sensitivity to apoptotic signaling, yet clinically relevant molecular actors to target mitochondria-mediated resistance remain elusive. Here, we show that MYC-driven neuroblastoma cells rely on intact mitochondrial ribosome (mitoribosome) processivity and undergo cell death following pharmacological inhibition of mitochondrial translation, regardless of their multidrug/mitochondrial resistance and stem-like phenotypes. Mechanistically, inhibiting mitoribosomes induced the mitochondrial stress-activated integrated stress response (ISR), leading to downregulation of c-MYC/N-MYC proteins prior to neuroblastoma cell death, which could be both rescued by the ISR inhibitor ISRIB. The ISR blocks global protein synthesis and shifted the c-MYC/N-MYC turnover toward proteasomal degradation. Comparing models of various neuroectodermal tumors and normal fibroblasts revealed overexpression of MYC proteins phosphorylated at the degradation-promoting site T58 as a factor that predetermines vulnerability of MYC-driven neuroblastoma to mitoribosome inhibition. Reducing N-MYC levels in a neuroblastoma model with tunable MYCN expression mitigated cell death induction upon inhibition of mitochondrial translation and functionally validated the propensity of neuroblastoma cells for MYC-dependent cell death in response to the mitochondrial ISR. Notably, neuroblastoma cells failed to develop significant resistance to the mitoribosomal inhibitor doxycycline over a long-term repeated (pulsed) selection. Collectively, we identify mitochondrial translation machinery as a novel synthetic lethality target for multidrug-resistant MYC-driven tumors.

• MeSH
• Apoptosis MeSH
• Humans MeSH
• Cell Line, Tumor MeSH
• Neuroblastoma * drug therapy   genetics   metabolism   MeSH
• N-Myc Proto-Oncogene Protein genetics   metabolism   MeSH
• Proto-Oncogene Proteins c-myc genetics   metabolism   MeSH
• Signal Transduction MeSH
• Synthetic Lethal Mutations * MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• N-Myc Proto-Oncogene Protein MeSH
• Proto-Oncogene Proteins c-myc MeSH

The aim of this work was to describe the pathotypes of Escherichia coli strains isolated from one-day-old chickens, as well as the occurrence of resistance and multidrug resistance (MDR) in these strains. A total of 429 mixed swabs from 4290 one-day-old chicks were examined between August 2021 and July 2023 (24 months) during routine point-of-destination inspections at 12 poultry farms in the Czech Republic. All samples were processed via cultivation methods using meat-peptone blood agar and Mc Conkey agar under aerobic conditions at 37 ± 1 °C for 18-24 h. The identification of the strains was performed using MALDI-TOF mass spectrometry. All confirmed strains of E. coli were screened via single or multiplex PCRs for the presence of genes encoding the virulence-associated factors iroN, cvaC, iss, felA, iutA, frz and tsh. Antimicrobial susceptibility tests were performed using the minimal inhibitory concentration (MIC) method, focusing on ampicillin, cefotaxime, tetracycline, doxycycline, enrofloxacin, florfenicol, amoxicillin with clavulanic acid and trimethoprim with sulfamethoxazole. A total of 321 E. coli strains (prevalence of 74.8%) were isolated, and 300 isolates were defined as avian pathogenic strains of E. coli (APEC) via multiplex PCR. Based on the defined virulence genes, the isolates were classified into 31 pathotypes. A total of 15.9% of the tested isolates were susceptible to all the tested antimicrobials. On the other hand, 20.5% of the isolates were identified as multidrug-resistant (8.7% of isolates were resistant to three antimicrobials, 7.3% to four antimicrobials, 3.6% to five antimicrobials and 0.9% to six antimicrobials). Monitoring pathogenic strains of E. coli in different animals and in the environment makes it possible to understand their spread in animal and human populations and, at the same time, reveal the sources of virulence and resistance genes.

• Keywords
• avian pathogenic E. coli, multidrug resistance, pathogenicity, poultry, prevention, virulence,
• Publication type
• Journal Article MeSH

Pituitary hormones play a central role in shaping vertebrate life history events, including growth, reproduction, metabolism, and aging. The regulation of these traits often requires precise control of hormone levels across diverse timescales. However, fine tuning circulating hormones in-vivo has traditionally been experimentally challenging. Here, using the naturally short-lived turquoise killifish (N. furzeri), we describe a high-throughput platform that combines loss- and gain-of-function of peptide hormones. Mutation of three primary pituitary hormones, growth hormone (gh1), follicle stimulating hormone (fshb), and thyroid stimulating hormone (tshb), alters somatic growth and reproduction. Thus, suggesting that while the killifish undergoes extremely rapid growth and maturity, it still relies on vertebrate-conserved genetic networks. As the next stage, we developed a gain-of-function vector system in which a hormone is tagged using a self-cleavable fluorescent reporter, and ectopically expressed in-vivo through intramuscular electroporation. Following a single electroporation, phenotypes, such as reproduction, are stably rescued for several months. Notably, we demonstrate the versatility of this approach by using multiplexing, dose-dependent, and doxycycline-inducible systems to achieve tunable and reversible expression. In summary, this method is relatively high-throughput, and facilitates large-scale interrogation of life-history strategies in fish. Ultimately, this approach could be adapted for modifying aquaculture species and exploring pro-longevity interventions.

In humans and other vertebrates, a pea-size gland at the base of the brain called the pituitary gland, produces many hormones that regulate how individuals grow, reproduce, and age. Three of the most prominent hormones are known as the growth hormone, the follicle-stimulating hormone, and the thyroid-stimulating hormone. It is important that the body precisely controls the levels of these hormones throughout an individual’s life. One way researchers can investigate how hormones and other molecules work is to artificially alter the levels of the molecules in living animals. However, this has proved to be technically challenging and time-consuming for pituitary gland hormones. Moses et al. studied the growth hormone, follicle-stimulating hormone, and thyroid-stimulating hormone in the turquoise killifish, a small fish that grows and matures more rapidly than any other vertebrate research model. The experiments revealed that mutant fish lacking one of the three primary pituitary hormones were smaller, took longer to reach maturity, or were completely sterile. This suggests these three hormones play a similar role in killifish as they do in other vertebrates. The team then developed a new experimental platform to precisely control the levels of the three hormones in killifish. Genes encoding individual hormones were expressed in the muscles of the mutant fish, effectively making the muscles a ‘factory’ for producing that hormone. Treating mutant fish this way once was enough to restore growth and to fully return reproduction to normal levels for several months. Moses et al. also demonstrated that it is possible to use this platform to express more than one hormone gene at a time and to use drugs to switch hormone production on and off in a reversible manner. For example, this reversible approach made it possible to effectively adjust fertility levels. The new platform developed in this work could be adapted for modifying a variety of traits in animals to explore how they impact health and longevity. In the future, it may also have other applications, such as optimizing how farmed fish grow and reproduce and regulating hormone levels in human patients with hormone imbalances.

• Keywords
• CRISPR, aging, aquaculture, developmental biology, genome editing, nothobranchius furzeri, peptide hormones, reproduction, somatic growth,
• MeSH
• Longevity MeSH
• Fundulidae * MeSH
• Pituitary Hormones MeSH
• Peptide Hormones * MeSH
• Growth Hormone metabolism   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Pituitary Hormones MeSH
• Peptide Hormones * MeSH
• Growth Hormone MeSH

While the prudent and reasonable use of veterinary antimicrobial agents in food-producing animals is necessary, researchers over the decades have shown that these antimicrobial agents can spread into the environment through livestock manure and wastewater. The analysis of the occurrence of antimicrobial compounds in soil samples is of a great importance to determine potential impacts on human and animal health and the environment. In this study, an affordable, rugged and simple analytical method has been developed for the determination of twenty-nine antimicrobial compounds from five different classes (tetracyclines, fluoro(quinolones), macrolides, sulfonamides and diaminopirimidines). Liquid-liquid extraction (LLE) with extract filtration combined with ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was the best strategy for the simultaneous determination of all analytes. The developed method was validated according to the Commission Implementing Regulation (EU) 2021/808. The limit of detections (LODs) ranged from 0.5 to 2.0 µg/kg, while the limit of quantitation (LOQ) was established at 1.0 to 20.0 µg/kg. The developed method was successfully applied for the determination of antimicrobial residues in one hundred and eighteen soil samples obtained from four European countries (Austria, Czech Republic, Estonia and Portugal). Doxycycline in the concentration levels of 9.07 µg/kg-20.6 µg/kg was detected in eight of the analysed samples. Samples were collected from areas where natural fertilizers (swine or cow manure) were applied. Our method can be efficiently used to monitor anti-microbial compounds in soil samples.

• Keywords
• UHPLC-MS/MS, antimicrobial agents, environment, soil,
• MeSH
• Anti-Bacterial Agents analysis   MeSH
• Anti-Infective Agents * MeSH
• Solid Phase Extraction MeSH
• Manure analysis   MeSH
• Humans MeSH
• Swine MeSH
• Soil MeSH
• Cattle MeSH
• Tandem Mass Spectrometry * methods   MeSH
• Chromatography, High Pressure Liquid MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Cattle MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Anti-Infective Agents * MeSH
• Manure MeSH
• Soil MeSH

A 2-year-old, spayed female, Bichon Frise dog was presented with reluctance to exercise, back pain, and frequent sitting down. Multiple osteolysis, periosteal proliferation, and sclerosis of the vertebral endplates of T11-13 were observed in the radiography, computed tomography, and magnetic resonance imaging. The bacterial culture of the urine specimen, the polymerase chain reaction (PCR) of the blood, and the antibody tests were positive for Brucella canis. Accordingly, discospondylitis caused by B. canis was diagnosed and doxycycline was administered. The clinical signs resolved and the culture and PCR results were negative afterwards. Doxycycline was discontinued after 6 months. The clinical signs recurred 2 weeks later, and the combination treatment of doxycycline and enrofloxacin was initiated. Though no clinical signs were observed after 9 months and the bacterial cultures and PCR were negative, the antibody titre remained at 1 : 200 or more. The dog will continue taking antibiotics until the antibody titre drops. To the best of our knowledge, this is the first case report of a clinical infection of B. canis associated with canine discospondylitis in the Republic of Korea. Although the clinical signs of brucellosis might improve with antibiotic treatment, the disease cannot be cured due to Brucella's various strategies to evade host immune systems. Specifically, it can proliferate and replicate within the host cells, resulting in an environment that makes treatment less effective. Furthermore, owing to its zoonotic potential, owners and veterinarians should consider lifelong management or euthanasia.

• Keywords
• Brucella canis, discospondylitis, doxycycline, enrofloxacin, zoonotic disease,
• Publication type
• Journal Article MeSH
• Case Reports MeSH

INTRODUCTION: Surgical treatment is associated with an unwanted response of the organism to the so-called surgical trauma. This response is called surgical stress. Ischaemia-reperfusion injury is one of essential causes of tissue damage. It comprises functional and structural changes in tissue that occur after the restoration of circulation, after an episode of ischaemia. Necrosis of irreversibly changed cells and endothelial and mitochondrial-induced tissue swelling occur. METHODS: Physiology, pathophysiology of endothelial glycocalyx: Endothelial glycocalyx is a 0.2 to 5 micrometres thin heteropolysaccharide layer that covers the endothelium on its intraluminal side. Backbone molecules of the glycocalyx include proteoglycans, glycoproteins, and glycosaminoglycans. Damage of the endothelial glycocalyx was described in trauma patients, in patients with septic shock, in ischemia and reperfusion injury, and during extensive surgical procedures. Approaches to prevent endothelial glycocalyx damage: Remote ischemic preconditioning was tested as a method of ischemia and reperfusion injury prevention during and after surgery. Nevertheless, the expected effect was not confirmed in performed meta-analyses. Endothelial glycocalyx damage can be prevented pharmacologically with a broad spectrum of substances, such as antithrombin III, doxycycline, hydrocortisone, etanercept, or nitric oxide donors. Hydrogen inhalation or albumin affects glycocalyx positively. Sulodexide provides a positive effect on the protection and reparation of endothelial glycocalyx. This proteoglycan with antithrombotic, fibrinolytic, hypofibrinogenemic, and lipolytic function is used for the treatment of venous diseases, ischaemic heart disease, and peripheral arterial disease. A positive effect of sulodexide on renal dysfunction was documented in a model of ischaemia and reperfusion injury. Equally, a positive effect of sulodexide was described on endothelium repair after its mechanical damage. CONCLUSION: Further research needs to be performed to evaluate the effect of endothelium-protectives on glycocalyx damage prevention and repair in ischaemia and reperfusion models involving large laboratory animals or in clinical trials in patients undergoing surgical revascularisation procedures.

• MeSH
• Endothelium, Vascular MeSH
• Glycocalyx * physiology   MeSH
• Ischemia MeSH
• Humans MeSH
• Reperfusion Injury * MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH