Growth Of Bacterial Cultures Research Articles

Microbial Isolates and Antimicrobial Resistance Patterns in Adults with Inborn Errors of Immunity: A Retrospective Longitudinal Analysis of Sputum Cultures

Introduction: Individuals with inborn errors of immunity (IEI) are at increased risk of respiratory infection and frequently receive prolonged broad-spectrum antibiotics, leading to antibiotic resistance. The aim of this study was to identify respiratory pathogens and antibiotic resistance patterns in IEI patients. Methods: We retrospectively studied 36 IEI patients with positive bacterial growth in sputum cultures between 2014 and 2023. Data covered hospitalizations, respiratory infections, yearly antibiotic prescriptions, past sputum cultures, and antibiotic sensitivities. Patients with primary ciliary dyskinesia (PCD) and bronchiectasis served as a control group. Results: A total of 314 sputum cultures were analyzed from patients with IEI, alongside 585 cultures from individuals with PCD and 113 cultures from patients with bronchiectasis. Patients with IEI had a median age of 23.5 years, with 61% male participants. The study compared the differences in bacterial isolates from sputum cultures and antibiotic resistance between patients with IEI and the control groups. The most common bacterial isolates across all groups were Haemophilus influenzae (159 isolates in IEI vs. 314 in PCD and 26 in bronchiectasis), Pseudomonas aeruginosa, and Streptococcus pneumoniae. In IEI patients, 992 symptomatic respiratory exacerbations and 43 pneumonia-related hospitalizations were recorded. Notably, H. influenzae in IEI patients showed high resistance rates to cefuroxime (82%), amoxicillin/clavulanic acid (66%), trimethoprim/sulfamethoxazole (59%), and ampicillin/sulbactam (49%). P. aeruginosa in IEI patients displayed significant resistance to ciprofloxacin (85%), ceftazidime (42%), and aminoglycosides (23–33%). Additionally, all S. pneumoniae isolates in IEI patients were tetracycline resistant, with high resistance rates to penicillin, clindamycin, and erythromycin. It is essential to highlight the substantial resistance of common pathogens to oral antibiotics. In contrast, the control groups exhibited lower resistance rates across all bacterial isolates. Conclusion: Antimicrobial resistance is a growing concern among vulnerable IEI patients. We suggest conducting similar investigations in other regions to address this issue. The findings should inform future infection management guidelines for IEIs.

Prevalence of Bacterial Urinary Tract Infection among Pregnant Women in Wasit Province in Iraq

Background: Urinary tract infections (UTIs) are the most common type of infection during pregnancy. UTI include a spectrum of disorders, ranging from those affecting the lower urinary tract, like asymptomatic bacteriuria and cystitis, to those affecting the kidney, such as pyelonephritis. In pregnancy UTI can lead to preterm labor, anemia and septicemia. Escherichia coli is the most common pathogenic microorganisms associated with UTI and counting up to 60–80% of all UTIs. Objective: The aim of this study is to determine the prevalence of urinary tract infection (UTI) among pregnant women in wasit province. Materials and Methods: Mid-stream urine samples from 100 pregnant women who attending to Al- Shaheed Fairuz hospital and private clinics in wasit province between the periods of March 2024 to July 2024 with the age range of (16-35) years. The urine samples were cultured on MacConkey and blood agars then examined by Macroscopic and Microscopic examination by Gram's stain. The colonies were examined by biochemical tests (Oxidase, catalase, manitol salt agar, novobiocin tests, IMViC) and confirmatory tests (API 20E and API Staph) for isolates. Results: The results showed that the percentage of bacterial growth culture among the pregnant women was 33%. Gram negative bacteria were the higher percentage with (73%). Eschirishia coli bacteria were the most frequently isolated organism (40%) while, the lower percentage for Streptococcus spp. (3%). UTI in pregnant women was commonest in the age group 20-24 years (42%). According to trimesters of pregnancy, the highest percentage of UTI was in third trimester (43%) followed by second trimester (39%), while the the first trimest was the the lowest percentage with (18%). Conclusions: Urinary tract infections (UTIs) are common during pregnancy. Untreated UTIs can lead to complications such as preterm labor, low birth weight and sepsis. E.coli is the predominant pathogen presently causing urinary tract infections at all stages of pregnancy. Urine culture and sensitivity tests are mandatory for all pregnant women during the different trimesters.

Formulation of Dual-Functional Nonionic Cetomacrogol Creams Incorporated with Bacteriophage and Human Platelet Lysate for Effective Targeting of MDR P. aeruginosa and Enhanced Wound Healing.

Successful development of phage-based therapeutics and their utility predominantly depend on the mode and route of phage administration. Topical and site-directed phage application evokes minimal immune clearance and allows more phage-host adsorption, thereby ensuring higher phage efficacy. However, a notable drawback of conventional topical phage applications is the absence of sustained release. Occlusive emollients guarantee the controlled release of active pharmaceutical ingredients (APIs), thereby facilitating administration, preventing moisture loss, and acting as a skin barrier. In this study, we developed phage and human platelet lysate (h-PL) incorporated cetomacrogol-based creams for combined phage therapy and wound healing. The base material for phage immobilization was formulated by emulsifying paraffin and sterile water with cetomacrogol (emulsifying agent). Specifically, we incorporated a Pseudomonas aeruginosa-infecting lytic phage vB_PaeM_M12PA in the formulation and characterized its genome in this study. Cetomacrogol, a nonionic PEG (polyethylene glycol) based ether, rendered phage stability and allowed initial burst release followed by continuous controlled release of phages from the embedding matrix in the initial 6-8 h. Rheological studies showed that the material has elastic properties with storage moduli (G') values ranging from 109.51 ± 2.10 to 126.02 ± 3.13 kPa, indicating frequency-independent deformation. Platelet lysates in the cream acted as wound healing agents, and in vitro evaluation of cell migration and wound healing capacity of h-PL showed a significant enhancement by the sixth hour compared to untreated groups. The phage-incorporated cream showed sustained phage release in solid media and a significant reduction in bacterial growth in liquid cultures. In vivo wound healing studies in 6-week-old Wistar rats with full-thickness excision wounds and subsequent histopathological studies showed that the formulation enhanced wound healing and tissue restoration efficiency. In conclusion, the study unveils a promising approach for integrated phage therapy and wound healing strategies.

FBS-based cryoprotective compositions for effective cryopreservation of gut microbiota and key intestinal microorganisms

ObjectiveThe need for innovative techniques to preserve microbiota for extended periods, while maintaining the species composition and quantitative balance of the bacterial community, is becoming increasingly important. To address this need, we propose an efficient approach to cryopreserve human gut microbiota using a two-component cryoprotective composition comprising fetal bovine serum (FBS) and 5% dimethyl sulfoxide (DMSO). Fetal serum is a commonly utilized component in the freezing media for eukaryotic cells, however, its effects on prokaryotic cells have not been extensively researched.ResultsIn our study, we demonstrated the high efficiency of using a two-component cryoprotective medium, FBS + 5% DMSO, for cryopreservation of human gut microbiota using three different methods. According to the obtained results, the intact donor microbiota was preserved at a level of 85 ± 4% of the initial composition based on fluorescent analysis using the LIVE/DEAD test. No differences in survival were observed when comparing with pure DMSO and FBS media. The photometric measurement method for growth of aerobic bacteria (A. johnsoni), facultative anaerobes (E. coli, E. faecalis), microaerophilic (L. plantarum), and obligate anaerobic bacterial cultures (E. barkeri, B. breve) also demonstrated high viability rates of 94–98% in the two-component protective medium, reaching intact control levels. However, for anaerobic microflora representatives, serum proved to be a more suitable cryoprotectant. Also, we demonstrated that using cryoprotective media with 50–75% FBS content is enough to preserve a significant level of bacterial cell viability, from an economic standpoint.

Predictive Significance of Laboratory Tests in Bacteremic Brucellosis.

Brucellosis is one of the most common zoonotic infections. Although culture is the gold standard diagnostic method, bacterial growth in blood cultures may not always occur due to various factors. We aimed to investigate demographic, clinical, and laboratory findings that may have predictive significance for bacteremia in brucellosis. Patients older than 18 years of age followed up with a diagnosis of brucellosis between 2012 and 2022 were included in this retrospective multicenter study. They were divided into two main subgroups according to their Brucella species reproductive status as bacteremic and non-bacteremic. A total of 743 patients, 370 (49.80%) bacteremic and 373 (50.20%) non-bacteremic brucellosis patients, were enrolled. The mean age of the bacteremic group (36.74 years) was lower than the non-bacteremic group (43.18 yr). High fever, chills/cold, sweating, nausea, vomiting, and weight loss were more common in the bacteremic group. In the bacteremic group, white blood cell count, platelet count, hemoglobin level, mean platelet volume, eosinophil, and neutrophil counts were lower, and lymphocyte, erythrocyte sedimentation rate, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and ferritin levels were higher. According to the receiver operating characteristic (ROC) analysis, when the cut-off value of ferritin was considered 67, it was the parameter with the strongest predictive significance in Brucella bacteremia. High ferritin level, low eosinophil count, and increased erythrocyte sedimentation rate were determined as the most critical laboratory findings in predicting bacteremia in brucellosis.

Mortality in hospitalized SARS-CoV-2 patients with contemporaneous bacterial and fungal infections.

The interplay between SARS-CoV-2 and contemporaneous bacterial or fungal culture growth may have crucial implications for clinical outcomes of hospitalized patients. This study aimed to quantify the effect of microbiological culture positivity on mortality among hospitalized patients with SARS-CoV-2. In this retrospective cohort study, we included adult hospitalized patients from OPTUM COVID-19 specific data set, who tested positive for SARS-CoV-2 within 14 days of hospitalization between 01/20/2020 and 01/20/2022. We examined outcomes of individuals with organisms growing on cultures from the bloodstream infections (BSIs), urinary tract, and respiratory tract, and a composite of the three sites. We used propensity score matching on covariates included demographics, comorbidities, and hospitalization clinical parameters. In a sensitivity analysis, we included same covariates but excluded critical care variables such as length of stay, intensive care unit stays, mechanical ventilation, and extracorporeal membrane oxygenation. The cohort included 104,560 SARS-CoV-2 positive adult hospitalized patients across the United States. The unadjusted mortality odds increased significantly with BSIs (98.7%) and with growth on respiratory cultures (RC) (176.6%), but not with growth on urinary cultures (UC). Adjusted analyses showed that BSIs and positive RC independently contribute to mortality, even after accounting for critical care variables. In SARS-CoV-2-positive hospitalized patients, positive bacterial and fungal microbiological cultures, especially BSIs and RC, are associated with an increased risk of mortality even after accounting for critical care variables associated with disease severity. These findings underscore the importance of stringent infection control and the effective management of secondary infections to improve patient outcomes.

Макроміцети Національного природного парку «Гуцульщина» та їх антибактеріальні властивості

The ability of ethanol extracts of 20 species of macromycetes to inhibit the growth of bacterial cultures was studied. Their influence on indicators of superoxide dismutase and catalase activities in relation to three types of test cultures of microorganisms was established. Extracts of Cantharellus cibarius, Grifola frondosa Laetiporus sulphureus, Leccinum scabrum, Mycena leptocephala, Russula rosea, Strobilomyces strobilaceus showed the largest diameters of growth retardation of microorganisms of test cultures. The ability to counteract bacteria is a species-specific feature and does not depend on the trophic timing of macromycetes, the same applies to the ability to suppress the catalase activity of microorganisms. Extracts of macromycetes, which provoked inhibition of the growth of cultures, led to changes in the SOD activity of the studied bacteria: during incubation of M. luteus culture with mushroom extracts, which showed an inhibitory effect on the growth of microorganisms with a diameter of 8-12 mm, superoxide dismutase activity increased 2-3 times, during incubation of M. luteus bacteria with extracts that inhibited growth with a diameter greater than 12 mm, the enzymatic activity increased 8-10 times. The SOD activity of B. spizizeni increased by 40-80% at the 24th hour of the experiment with extracts, the use of which is accompanied by the formation of a 10 mm zone of growth inhibition, but already after 48 hours of the experiment, the indicators of the SOD activity of B. spizizeni decreased to the control values. SOD activity of E. coli increased on the 24th hour of the experiment by 1.4-1.6 times relative to the control values, even with the use of extracts that provoked inhibition of the growth of cultures with a diameter of 8 mm and remained high on the 48th hour of the experiment (in 1.2-1.3 times).

A NEW ROLE FOR KYNURENINE METABOLISM IN THE AGE-ASSOCIATED DECLINE OF THE BACTERIAL IMMUNE RESPONSE

Abstract Tryptophan metabolism through the kynurenine pathway becomes dysregulated during normal aging and is implicated in age-associate disease, including chronic inflammation, atherosclerosis, neurodegeneration, and cancer. Kynurenine pathway enzymes and metabolites influence a range of molecular processes critical to healthy aging, including regulation of inflammatory and immune responses. Kynurenine metabolism is active in immune cells and activated in response to proinflammatory cytokine signaling. We previously determined that elevating physiological levels of the kynurenine pathway metabolite 3-hydroxyanthranilic acid (3HAA) via either direct supplementation or inhibition of the enzyme that degrades 3HAA, 3HAA dioxygenase (HAAO), extends lifespan and delays age-associated health decline in Caenorhabditis elegans. Published work suggests that 3HAA is anti-inflammatory in mammals, for example by reducing the ratio of activated to regulatory T cells and inhibiting pathological activation of macrophages. In recent work, we find that elevating physiological 3HAA can beneficially enhance the immune response of C. elegans to bacterial pathogens during aging. 3HAA is sufficient to kill bacterial and inhibit bacterial growth in culture. When HAAO is inhibited in C. elegans, 3HAA accumulates in lysosome related organelles (LROs) in the intestinal cells, the same subcellular compartment that contains engulfed bacteria. LROs are a cellular repository for both iron and zinc, and we further find that iron chelation or zinc supplementation dramatically enhances the bactericidal properties of 3HAA. Here we present a mechanistic model in which age-dependent accumulation of 3HAA in intestinal LROs combines with zinc to enhance bacterial resistance with age in C. elegans.

A phenomenological model of non-genomic variability of luminescent bacterial cells

The light emitted by a luminescent bacterium serves as a unique native channel of information regarding the intracellular processes within the individual cell. In the presence of highly sensitive equipment, it is possible to obtain the distribution of bacterial culture cells by the intensity of light emission, which correlates with the amount of luciferase in the cells. When growing on rich media, the luminescence intensity of individual cells of brightly luminous strains of the luminescent bacteria Photobacterium leiognathi and Ph. phosporeum reaches 104–105 quanta/s. The signal of such intensity can be registered using sensitive photometric equipment. All experiments were carried out with bacterial clones (genetically homogeneous populations). A typical dynamics of luminous bacterial cells distributions with respect to intensity of light emission at various stages of batch culture growth in a liquid medium was obtained. To describe experimental distributions, a phenomenological model that links the light of a bacterial cell with the history of events at the molecular level was constructed. The proposed phenomenological model with a minimum number of fitting parameters (1.5) provides a satisfactory description of the complex process of formation of cell distributions by luminescence intensity at different stages of bacterial culture growth. This may be an indication that the structure of the model describes some essential processes of the real system. Since in the process of division all cells go through the stage of release of all regulatory molecules from the DNA molecule, the resulting distributions can be attributed not only to luciferase, but also to other proteins of constitutive (and not only) synthesis.

Renal Function Impairment in Children With Congenital Cytomegalovirus Infection: A Cross-sectional Study.

We aimed to determine the prevalence and severity of glomerular and tubular renal dysfunction by means of urinalysis in infants and toddlers with congenital cytomegalovirus infection (cCMV) and their association with cCMV disease, viruria and antiviral treatment. This cross-sectional study was done using the Spanish Registry of Congenital Cytomegalovirus Infection. First-morning urine samples were collected from January 2016 to December 2018 from patients <5 years old enrolled in Spanish Registry of Congenital Cytomegalovirus Infection. Samples were excluded in case of fever or other signs or symptoms consistent with acute infection, bacteriuria or bacterial growth in urine culture. Urinary protein/creatinine and albumin/creatinine ratios, urinary beta-2-microglobulin levels, hematuria and CMV viruria were determined. A 0.4 cutoff in the urinary albumin/protein ratio was used to define tubular (<0.4) or glomerular (>0.4) proteinuria. Signs and symptoms of cCMV at birth, the use of antivirals and cCMV-associated sequelae at last available follow-up were obtained from Spanish Registry of Congenital Cytomegalovirus Infection. Seventy-seven patients (37 females, 48.1%; median [interquartile range] age: 14.0 [4.4-36.2] months) were included. Symptom-free elevated urinary protein/creatinine and albumin/creatinine ratios were observed in 37.5% and 41.9% of patients, respectively, with tubular proteinuria prevailing (88.3%) over glomerular proteinuria (11.6%). Proteinuria in the nephrotic range was not observed in any patients. In multivariate analysis, female gender was the only risk factor for tubular proteinuria (adjusted odds ratio = 3.339, 95% confidence interval: 1.086-10.268; P = 0.035). cCMV disease at birth, long-term sequelae, viruria or the use of antivirals were not associated with urinalysis findings. Mild nonsymptomatic tubular proteinuria affects approximately 40% of infants and toddlers with mostly symptomatic cCMV in the first 5 years of life.

Mathematical modeling of quorum sensing dynamics in batch culture of luminescent bacterium Photobacterium phosphoreum 1889.

At the beginning of the paper, the level of necessary phenomenology of complex models is discussed. When working with complex systems, which of course include living organisms and ecological systems, it is necessary to use a phenomenological description. An illustration of the phenomenological approach is given, which captures the most significant general principles or patterns of interactions; the specific values of the parameters cannot be calculated from the first principles, but are determined empirically. An appropriate interpretation is also chosen empirically and pragmatically. However, in order to simulate a wider range of situations, it becomes necessary to lower the level of phenomenology, switch to a more detailed description of the system, introducing interaction between selected elements of the system. The requirements for a system model combining ecological, metabolic and genetic levels of cell culture description are formulated. A mathematical model of quorum sensing dynamics during the growth of batch culture of luminescent bacteria at different concentrations of the nutrient substrate has been developed. The model contains four blocks describing ecological, energy, quorum and luminescent aspects of bacterial culture growth. The model demonstrated good agreement with the experimental data obtained. When analyzing the model, three oddities in the behavior of the culture were noted, which presumably can change the idea of some processes taking place during the development of a culture of luminescent bacteria. The results obtained suggest the presence of some additional control system for the luminescent reaction via the synthesis pathways of FMN · Н2 or aliphatic aldehyde. In this case, the generalized description of the contribution of energy metabolism to luminescence only through ATP is too strong a simplification. As a result of comparing the model dynamics with the experiment, a discrepancy arose between the concentration of the substrate (peptone) measured in the experiment and its effective influence on the bacterial population growth. This discrepancy seems to indicate peptone is not the leading substrate, and growth is limited by nutrients contained in the yeast extract, the concentration of which did not change in these experiments. The discrepancies noted between the expectations and the results of experimental data processing, together with the assumptions about the causes of these discrepancies, set the direction for further experimental and theoretical studies of quorum sensing mechanisms in a culture of luminescent bacteria.

The role of microbiota in tumorigenesis, progression and treatment of bladder cancer.

For decades, the urinary system was regarded as a sterile environment due to the absence of any bacterial growth in clinical standard urine cultures from healthy individuals. However, a diverse array of microbes colonizes the urinary system in small quantities, exhibiting a variable compositional signature influenced by differences in sex, age, and pathological state. Increasing pieces of evidence suggest microbiota exists in tumor tissue and plays a crucial role in tumor microenvironment based on research in multiple cancer models. Current studies about microbiota and bladder cancer have preliminarily characterized the bladder cancer-related microbiota, but how the microbiota influences the biological behavior of bladder cancer remains unclarified. This review summarizes the characteristics of microbiota in bladder cancer, aims to propose possible mechanisms that microbiota acts in tumorigenesis and progression of bladder cancer based on advances in gut microbiota, and discusses the potential clinical application of microbiota in bladder cancer.

Deep learning enhanced multiplex detection of viable foodborne pathogens in digital microfluidic chip

Culture plating is worldwide accepted as the gold standard for quantifying viable foodborne pathogens. However, it is time-consuming (1–2 days) and requires specialized laboratory and personnel. This study reported a deep learning enhanced digital microfluidic platform for multiplex detection of viable foodborne pathogens. The new method used a Time-Lapse images driven EfficientNet-Transformer Network (TLENTNet) to type and quantify the bacteria through spatiotemporal features of bacterial growth and digital enumeration of bacterial culture. First, the bacterial sample was prepared with LB medium and injected into a pre-vacuumed microfluidic chip with an array of 800 microwells to encapsulate at most one bacterium in each well. Then, a programmed sliding microscopic platform was used to scan all microwells every 15 min, capturing time-lapse images of bacterial growth within each microwell. Finally, the TLENTNet was used to facilitate bacterial typing and quantification. Under optimal conditions, this platform was able to detect four bacterial species (S.typhimurium, E. coli O157:H7, S. aureus and B. cereus) with an average accuracy of 97.72% and a detection limit of 63 CFU/mL in 7 h.

Thermal adaptation of soil microbial growth traits in response to chronic warming.

Soils are the largest terrestrial carbon sink and the foundation of our food, fiber, and fuel systems. Healthy soils are carbon sinks, storing more carbon than they release. This reduces the amount of carbon dioxide released into the atmosphere and buffers against climate change. Soil microbes drive biogeochemical cycling and contribute to soil health through organic matter breakdown, plant growth promotion, and nutrient distribution. In this study, we determined how soil microbial growth traits respond to long-term soil warming. We found that bacterial isolates from warmed plots showed evidence of adaptation of optimum growth temperature. This suggests that increased microbial biomass and growth in a warming world could result in greater carbon storage. As temperatures increase, greater microbial activity may help reduce the soil carbon feedback loop. Our results provide insight on how atmospheric carbon cycling and soil health may respond in a warming world.

Study of the prevalence and antibiotics susceptibility patterns in clinically significant isolates among urine sample in a tertiary care centre

Urinary tract infections (UTIs) are prevalent bacterial infections. Gram-negative bacilli are frequently isolated as the causative pathogens in UTIs, and antibiotics are commonly used to treat them. However, the overuse of antibiotics has led to the development of resistance in some uropathogens. : To find the prevalence and antimicrobial susceptibility patterns in clinically significant isolates among urine sample in a tertiary care centre.: Fresh mid-stream urine samples were collected aseptically in sterile containers. The samples were inoculated on 5% sheep blood agar and MacConkey agar. The isolates were identified to the species level using biochemical tests/Vitek 2 compact.: A total of 5847 specimens were examined for significant bacteriuria, out of which 1160 showed bacterial growth in urine cultures. The most commonly isolated pathogens were (57.67%) and (18.63%). The antibiotics aminoglycosides (84%), carbapenems (79%), and nitrofurantoin (63%) displayed a high rate of sensitivity against these pathogens.: Due to the variability of drug sensitivity among bacterial pathogens over time, regular surveillance and monitoring are essential to provide physicians with updated information for the most effective empirical treatment of UTIs.

Purification of linearized template plasmid DNA decreases double-stranded RNA formation during IVT reaction.

After the COVID-19 pandemic, messenger RNA (mRNA) has revolutionized traditional vaccine manufacturing. With the increasing number of RNA-based therapeutics, valuable new scientific insights into these molecules have emerged. One fascinating area of study is the formation of double-stranded RNA (dsRNA) during in vitro transcription (IVT) which is considered a significant impurity, as it has been identified as a major trigger in the cellular immune response pathway. Therefore, there is a growing importance placed to develop and optimize purification processes for the removal of this by-product. Traditionally, efforts have primarily focused on mRNA purification after IVT through chromatographic separations, with anion exchange and reverse phase chromatography emerging as effective tools for this purpose. However, to the best of our knowledge, the influence and significance of the quality of the linearized plasmid have not been thoroughly investigated. Plasmids production involves the growth of bacterial cultures, bacterial harvesting and lysis, and multiple filtration steps for plasmid DNA purification. The inherent complexity of these molecules, along with the multitude of purification steps involved in their processing, including the subsequent linearization and the less-developed purification techniques for linearized plasmids, often result in inconsistent batches with limited control over by-products such as dsRNA. This study aims to demonstrate how the purification process employed for linearized plasmids can impact the formation of dsRNA. Several techniques for the purification of linearized plasmids based on both, resin filtration and chromatographic separations, have been studied. As a result of that, we have optimized a chromatographic method for purifying linearized plasmids using monolithic columns with C4 chemistry (butyl chains located in the surface of the particles), which has proven successful for mRNAs of various sizes. This chromatographic separation facilitates the generation of homogeneous linearized plasmids, leading to mRNA batches with lower levels of dsRNA during subsequent IVT processes. This finding reveals that dsRNA formation is influenced not only by RNA polymerase and IVT conditions but also by the quality of the linearized template. The results suggest that plasmid impurities may contribute to the production of dsRNA by providing additional templates that can be transcribed into sequences that anneal with the mRNA molecules. This highlights the importance of considering the quality of plasmid purification in relation to dsRNA generation during transcription. Further investigation is needed to fully understand the mechanisms and implications of plasmid-derived dsRNA. This discovery could shift the focus in mRNA vaccine production, placing more emphasis on the purification of linearized plasmids and potentially saving, in some instances, a purification step for mRNA following IVT.

Patients with Cerebral Palsy Who are Admitted to a Chest Diseases Hospital may have Higher Mortality Rates Serebral Palsili Hastaların Göğüs Hastalıkları Hastanesine Yatışları Mortal Seyredebilir

Objective: Epilepsy, pneumonia, gastritis, esophagitis, malnutrition, dehydration, fractures, and non-pneumonic lung disorders (asthma, pleural fluid, respiratory failure) are the leading causes of hospitalization among cerebral palsy (CP) patients. This study aimed to determine respiratory tract diseases and mortality rates in patients with CP admitted to the chest diseases hospital. &#x0D; Materials and Methods: Patients diagnosed with CP and hospitalized in Chest Diseases Hospital between January 2010 and December 2017 were included in the study and analyzed retrospectively. As the endpoint, hospitalized patients' respiratory tract infection-related 30-days mortality were examined.&#x0D; Results: Forty-three hospitalizations of 23 patients were evaluated. The mean length of hospital stay was 15.5 (1-62) days. The reasons for hospitalization in patients with CP were mainly composed of respiratory tract infections (pneumonia %88,4, lung abscess %7). The most frequently isolated pathogenic microorganisms were gram-negative bacteria. Wheezing, bacterial culture growth, vasopressor and invasive mechanical ventilation were all factors that increased 30-day mortality (respectively p=0,016; p=0,024; p=0,003; p=0,003). The 30-day mortality rate was %21,7. While 2 patients died at first hospitalization, 4 patients died on recurrent hospitalizations. &#x0D; Conclusion: Patients with CP are often hospitalized due to respiratory tract infections. Recurrent hospitalizations may increase mortality. This study emphasizes the importance of close follow-up with these patients considering the high mortality rate among CP patients hospitalized for respiratory tract infections.

Single-cell mass distributions reveal simple rules for achieving steady-state growth.

Microbiologists have watched clear liquid turn cloudy for over 100 years. While the cloudiness of a culture is proportional to its total biomass, growth rates from optical density measurements are challenging to interpret when cells change size. Many bacteria adjust their size at different steady-state growth rates, but also when shifting between starvation and growth. Optical density cannot disentangle how mass is distributed among cells. Here, we use single-cell mass measurements to demonstrate that a population of cells in batch culture achieves a stable mass distribution for only a short period of time. Achieving steady-state growth in rich medium requires low initial biomass concentrations and enough time for individual cell mass accumulation and cell number increase via cell division to balance out. Steady-state growth is important for reliable cell mass distributions and experimental reproducibility. We discuss how mass variation outside of steady-state can impact physiology, ecology, and evolution experiments.

Whole-Genome Sequencing and Biotechnological Potential Assessment of Two Bacterial Strains Isolated from Poultry Farms in Belgorod, Russia.

Bacteria, designated as A1.1 and A1.2, were isolated from poultry waste based on the ability to form ammonia on LB nutrient medium. Whole genome sequencing identified the studied strains as Peribacillus frigoritolerans VKM B-3700D (A1.1) and Bacillus subtilis VKM B-3701D (A1.2) with genome sizes of 5462638 and 4158287 bp, respectively. In the genome of B. subtilis VKM B-3701D, gene clusters of secondary metabolites of bacillin, subtilisin, bacilisin, surfactin, bacilliacin, fengycin, sactipeptide, and ratipeptide (spore killing factor) with potential antimicrobial activity were identified. Clusters of coronimine and peninodin production genes were found in P. frigoritolerans VKM B-3700D. Information on coronimine in bacteria is extremely limited. The study of the individual properties of the strains showed that the cultures are capable of biosynthesis of a number of enzymes, including amylases. The B. subtilis VKM V-3701D inhibited the growth of bacterial test cultures and reduced the growth rate of the mold fungus Aspergillus unguis VKM F-1754 by 70% relative to the control. The antimicrobial activity of P. frigoritolerans VKM V-3700D was insignificant. At the same time, a mixture of cultures P. frigoritolerans VKM B-3700D/B. subtilis VKM B-3701D reduced the growth rate of A. unguis VKM F-1754 by 24.5%. It has been shown that strain A1.1 is able to use nitrogen compounds for assimilation processes. It can be assumed that P. frigoritolerans VKM V-3700D belongs to the group of nitrifying or denitrifying microorganisms, which may be important in developing methods for reducing nitrogen load and eutrophication.

Therapeutic-oligonucleotides activated by nucleases (TOUCAN): A nanocarrier system for the specific delivery of clinical nucleoside analogues

Nucleoside analogues have been in clinical use since 1960s and they are still used as the first therapeutic option for several cancers and viral infections, due to their high therapeutic efficacy. However, their wide clinical acceptance has been limited due to their high toxicity and severe side effects to patients. Herein, we report on a nanocarrier system that delivers nucleosides analogues in a target-specific manner, making nucleoside-based therapeutics safer and with the possibility to be used in other human conditions. This system, named, Therapeutic OligonUCleotides Activated by Nucleases” (TOUCAN) combines: i) the recognition power of oligonucleotides as substrates, ii) the use of nucleases as enzymatic biomarkers and iii) the clinical efficacy of nucleoside analogues, in a single approach. As a proof-of-concept, we report on a TOUCAN that is activated by a specific nuclease produced by bacteria and releases a therapeutic nucleoside, floxuridine. We demonstrate, for the first time, that, by incorporating a therapeutic nucleoside analogue into oligonucleotide probes, we can specifically inhibit bacterial growth in cultures. In this study, Staphylococcus aureus was selected as the targeted bacteria and the TOUCAN strategy successfully inhibited its growth with minimal inhibitory concentration (MIC) values ranging from 0.62 to 40 mg/L across all tested strains. Moreover, our results indicate that the intravenous administration of TOUCANs at a dose of 20 mg/kg over a 24-h period is a highly effective method for treating bacterial infections in a mouse model of pyomyositis. Importantly, no signs of toxicity were observed in our in vitro and in vivo studies. This work can significantly impact the current management of bacterial infections, laying the grounds for the development of a different class of antibiotics. Furthermore, it can provide a safer delivery platform for clinical nucleoside therapeutics in any human conditions, such as cancer and viral infection, where specific nuclease activity has been reported.