Colony-forming Units Research Articles

Targeted metabolomics analysis approach to unravel the biofilm formation pathways of Enterococcus faecalis clinical isolates.

(i) To characterize Enterococcus faecalis biofilm formation pathways by semi-targeted metabolomics and targeted nitrogen panel analysis of strong (Ef63) and weak (Ef 64) biofilm forming E. faecalis clinical isolates and (ii) to validate the identified metabolic markers using targeted inhibitors. Previous proteomics profiling of E. faecalis clinical isolates with strong and weak biofilm formation revealed that differences in metabolic activity levels of small molecule, nucleotide and nitrogen compound metabolic processes and biosynthetic pathways, cofactor metabolic process, cellular amino acid and derivative metabolic process and lyase activity were associated with differences in biofilm formation. Hence, semi-targeted analysis of Ef 63, Ef 64 and ATC control strain Ef 29212 was performed by selecting metabolites that were part of both the previously identified pathways and a curated library with confirmed physical and chemical identity, followed by confirmatory targeted nitrogen panel analysis. Significantly regulated metabolites (p < .05) were selected based on fold change cut-offs of 1.2 and 0.8 for upregulation and downregulation, respectively, and subjected to pathway enrichment analysis. The identified metabolites and pathways were validated by minimum biofilm inhibitory concentration (MBIC) and colony forming unit (CFU) assays with targeted inhibitors. Metabolomics analysis showed upregulation of betaine, hypoxanthine, glycerophosphorylcholine, tyrosine, inosine, allantoin and citrulline in Ef 63 w.r.t Ef 64 and Ef 29212, and thesemetabolites mapped to purinemetabolism, urea cycle and aspartate metabolism pathways. MBIC and CFU assays using compounds against selected metabolites and metabolic pathways, namely glutathione against hypoxanthine and hydroxylamine against aspartate metabolism showed inhibitory effects against E. faecalis biofilm formation. The study demonstrated the importance of oxidative stress inducers such as hypoxanthine and aspartate metabolism pathway in E. faecalis biofilm formation. Targeted therapeutics against these metabolic markers can reduce the healthcare burden associated with E. faecalis infections.

Potassium iodide enhances the killing effect of methylene blue mediated photodynamic therapy against F. monophora

BackgroundChromoblastomycosis (CMB) is a chronic granulomatous fungal infection that affect the skin and subcutaneous tissues. It is clinically problematic due to limited treatment options, low cure rates, and high rates of relapse. This underscores the necessity for innovative treatment approaches. In this study, potassium iodide (KI) combined with Methylene Blue (MB) mediated antimicrobial photodynamic therapy (PDT) were assessed in the treatment of Fonsecaea monophora (F. monophora) both in vitro and in vivo. And the underlying mechanism that contributes to the efficacy of this treatment approach was investigated. MethodsIn vitro experiments were conducted using different combinations and concentrations of MB, KI, and 660 nm light (60 mW/cm2) to inhibit F. monophora. The study was carried out using colony-forming unit (CFU) counts and scanning electron microscopy (SEM). The production of singlet oxygen (1O2), free iodine (I2), hydrogen peroxide (H2O2), and superoxide anion during the KI combined MB-mediated antimicrobial PDT process was also detected. In vivo experiments were developed using a Balb/c mouse paw infection model with F. monophora and treated with PBS, 10 mM KI, 2 mM MB +100 J/cm² and 10 mM KI+2 mM MB +100 J/cm² respectively. Inflammatory swelling, fungal load and histopathological analyses of the mouse footpads were assessed. ResultsKI enhanced the killing effect of MB-mediated antimicrobial PDT on the conidial spores of F. monophora at the cell and infected animal model level. During the process, the main antimicrobial agents in KI combined with MB- mediated antimicrobial PDT could produce stronger toxic active species including free I2 and H2O2. ConclusionKI combined with MB-mediated antimicrobial PDT could be an effective adjunct therapy for treating CBM.

Disinfection efficacy of slightly acidic electrolyzed water on microorganisms: Application in contaminated waste sorting rooms across varied scenarios

Various pathogenic microorganisms present in municipal solid waste (MSW) can pose significant hazards to both the environment and public health. Spraying Slightly Acidic Electrolyzed Water (SAEW) is an effective method for decontaminating microorganisms in the air and on surfaces in the waste sorting rooms. SAEW treatment, with an effective chlorine concentration (ECC) of 28.4–44.5 mg/L and a pH range of 5.0–6.5, was conducted in the waste sorting rooms of the old, high-rise, and villa residential communities in Shanghai, China. The efficacy of SAEW treatment was assessed by examining the microbial population in the air, on waste containers, on sanitation workers' hands, and the microbial community structure of MSW samples. SAEW treatment resulted in the total bacterial reduction ranging from 0.07 to 1.70 log10 colony-forming units (CFU)/cm2 on waste containers, with corresponding reduction efficiencies between 15.7% and 98.0%. Additionally, the positive rates of coliforms were decreased by 18.7%–95.2%. Furthermore, the application of SAEW demonstrated a significant impact on reducing airborne bacterial and fungal levels. SAEW was proven effective on object surfaces, in the air and on human skin, achieving disinfection efficiencies ranging from 22.7% to 54.3% on sanitation workers’ hands. And the most significant inhibition of microbial growth was observed in the waste sorting room of the villa residential community. The microbial diversity and structure of MSW samples exhibited noticeable differences before and after the SAEW application. These findings suggest that SAEW holds significant potential as an ideal, environmentally friendly broad-spectrum disinfectant for preventing and controlling microbial contamination in the waste sorting rooms.

Green Synthesis of Silver Nanoparticles Using Plant Extract Blends and Its Impact on Antibacterial and Biological Activity

There is a strong interest in using green resources for synthesizing nanoparticles (NPs) of industrial and biomedical utility in a way to maintain desired material properties throughout use while not inducing any harmful effects. The use of various plant extracts as reducing, capping, or stabilizing agents is widely attempted in green nanotechnology. However, very little has been explored about incorporating plant extract blends into green NP synthesis routes. Here, we used the combination of tea and olive leaf extracts for the synthesis of silver NPs and evaluated the advantages it provided over both chemical and single-plant-mediated synthesis routes. Four different reducing agents (tannic acid, black tea leaves extract, olive leaves extract and their blend) were used to synthesize silver NPs (Ag NP) from silver nitrate (AgNO[Formula: see text]. The synthesized Ag NP was characterized by scanning electron microscopy (SEM), dynamic light scattering (DLS), and ultraviolet-visible (US-Vis) spectroscopy. The antimicrobial properties of Ag NP were assessed against Escherichia coli (E. Coli) and Staphylococcus aureus (S. Aureus) using the colony-forming unit (CFU) assay and the minimum inhibitory concentration (MIC) assay. The cytotoxic potential of Ag NP on human colorectal adenocarcinoma (Caco-2) cells was assessed by the WST-1 assay. Results showed that Ag NP synthesized using plant extract mixtures had a primary particle size of 40[Formula: see text]nm and were very effective antibacterial agents, with the MIC values ranging from [Formula: see text]g/mL to 10[Formula: see text][Formula: see text]g/mL. While the particle size obtained in chemical synthesis was slightly lower, the resultant Ag NP did not serve as an effective antibacterial agents at low doses. Further understanding of how best to integrate extracts of different plants into green NP synthesis routes will enable wider and safer biomedical applications.

Analysis of a new phage, KZag1, infecting biofilm of Klebsiella pneumoniae: genome sequence and characterization

BackgroundThis study investigates the effectiveness of the bacteriophage KZag1 against drug-resistant Klebsiella pneumoniae, aiming to assess its potential as a therapeutic agent. The novelty lies in the characterization of KZag1, a Myovirus with specific efficacy against multidrug-resistant K. pneumoniae strains. This highlights the significance of exploring alternative strategies, particularly phage therapy, in addressing biofilm-associated infections.MethodsKZag1, characterized by a typical Myovirus structure with a 75 ± 5 nm diameter icosahedral head and a 15 ± 5 nm short tail, was evaluated in experimental trials against 15 strains of K. pneumoniae. The infection cycle duration was determined to be 50 min, resulting in an estimated burst size of approximately 83 plaque-forming units per colony-forming unit (PFU/CFU). Stability assessments were conducted within a pH range of 4 to 12 and temperatures ranging from 45°C to 60°C. Biofilm biomass reduction was observed, particularly at a multiplicity of infection (MOI) of 10.ResultsKZag1 demonstrated infection efficacy against 12 out of 15 tested K. pneumoniae strains. The phage exhibited stability across a broad pH range and at elevated temperatures. Notably, treatment with KZag1 significantly reduced K. pneumoniae biofilm biomass, emphasizing its potential in combating biofilm formation. Genomic analysis revealed a complete genome of 157,089 base pairs with a GC content of 46.38%, encompassing 203 open reading frames (ORFs) and a cysteine-specific tRNA sequence. Comparison with phage GP4 highlighted similarities, with KZag1 having a longer genome by approximately 4829 base pairs and a higher GC content by approximately 0.93%. Phylogenetic analysis classified KZag1 within the Myoviridae family.ConclusionThe efficacy of KZag1 against K. pneumoniae biofilm suggests its potential as a therapeutic candidate, especially for drug-resistant infections. Further clinical research is warranted to explore its synergy with other treatments, elucidate genomic traits, compare with Myoviridae phages, and understand its host interactions. These findings underscore the promising role of KZag1 in addressing drug-resistant bacterial infections.

Point-of-Care Electrochemical Immunosensor Applied against Nosocomial Infection: Staphylococcus aureus Detection in Human Hand Skin.

Staphylococcus aureus is an important pathogen that causes nosocomial infections, resulting in unacceptable morbidity and mortality rates. In this work, we proposed the construction of a nanostructured ZnO-based electrochemical immunosensor for qualitative and semiquantitative detection of S. aureus using simple methods for growing zinc oxide nanorods (ZnO NRs) on a sensor board and immobilizing the anti-S. aureus antibody on ZnO NRs through cystamine and glutaraldehyde. The immunosensor detected S. aureus in the 103-107 colony-forming unit (CFU) mL-1 range and showed a limit of detection (LoD) around 0.792 × 103 CFU mL-1. Beyond a satisfactory LoD, the developed immunosensor presented other advantages, such as high versatility for point-of-care assays and a suitable selective factor that admits the detection of the S. aureus concentration range in human hand skin after washing. Moreover, the immunosensor showed the potential to be an excellent device to control nosocomial infection by detecting the presence of S. aureus in human hand skin.

ANTIMICROBIAL EFFECT OF OZONIZED PHYSIOLOGICAL SOLUTION ON BACTERIA CAUSING CYSTITIS IN DOGS

Bacterial cystitis is a common clinical problem among dogs and is one of the reasons for the empirically administration of antimicrobials. This practice facilitates the selection of bacteria that are multidrug-resistant to antibiotics. In this context, it is urgent to understand and validate therapeutic modalities that complement antimicrobial treatment in cystitis cases. Ozone therapy has been proposed by scientists both in human and animal medicine. Thus, the objective of this study was evaluated the in vitro antimicrobial activity of ozonized saline solution over standard strains and resistant or multi-drug resistant isolates commonly associated with cystitis in dogs. The plating method was used to evaluate the antimicrobial activity of the exposure of 1 mL of phosphate buffer solution containing 108 colony forming units (CFU) mL-1 for 60 s to 4 mL of ozonized saline solution at 78 µg mL-1 over the standard strains (Staphylococcus aureus and Escherichia coli) and resistant or multi-resistant isolates (Proteus mirabillis, Klebsiella sp. e Enterococcus sp.), and the broth microdilution test to determine the minimum inhibitory and bactericidal concentration for S. aureus, E. coli and Pseudomonas aeruginosa strains from the same solution. The plating was the only assay in which treatment with ozonized saline resulted in bactericidal activity, promoting a reduction of more than 99% of colony-forming units for most of the tested strains, except for P. mirabillis. These results are promising, as these disease is the most common lower urinary tract disorder in dogs, with limited allopathic therapy due to increased bacterial resistance.

Limosilactobacillus reuteri 6475 and Prevention of Early Postmenopausal Bone Loss

Daily supplementation with the probiotic Limosilactobacillus reuteri ATCC PTA 6475 (L reuteri) vs placebo has previously been demonstrated to reduce bone loss in an estrogen deficiency mice model and older women, although the magnitude of the effect was small. We hypothesized that long-term treatment with L reuteri could result in clinically relevant skeletal benefits in postmenopausal osteoporosis. To evaluate whether daily supplementation with L reuteri vs placebo could reduce early postmenopausal bone loss and whether the effects remained or increased over time during 2 years of treatment. A double-blind, randomized, placebo-controlled clinical trial was conducted between December 4, 2019, and October 6, 2022, at a single center in Gothenburg, southwestern Sweden. Participants were recruited by online advertisements, and letters were sent to 10 062 women aged 50 to 60 years. Responding women (n = 752) underwent telephone screening, resulting in 292 women being invited to a screening visit. Of those who were screened, 239 women met all inclusion criteria and had no exclusion criteria. Capsules with L reuteri in 2 doses, 5 × 108 (low dose) or 5 × 109 (high dose) colony-forming units, taken twice daily or placebo were administered. All capsules also included cholecalciferol, 200 IU. The primary outcome was the relative change in tibia total volumetric bone mineral density (vBMD) over 2 years. Secondary outcomes included relative change in areal BMD of the lumbar spine and total hip, bone turnover markers C-terminal telopeptide cross-links of collagen type I and type I procollagen intact N-terminal propeptide, as well as tibia trabecular bone volume fraction and cortical vBMD. Both intention-to-treat and per-protocol analyses were conducted. A total of 239 postmenopausal women (median age, 55 [IQR, 53-56] years) were included. Tibia vBMD (primary outcome), hip and spine vBMD, and tibia cortical area and BMD decreased significantly in all groups, with no group-to-group differences (percent change tibia vBMD high dose vs placebo least-squares means, -0.08 [95 CI, -0.85 to 0.69] and low dose vs placebo least-squares means, -0.22 [95% CI, -0.99 to 0.55]). There were no significant treatment effects on any other predefined outcomes. A prespecified sensitivity analysis found a significant interaction between body mass index (BMI) and treatment effect at 2 years. No significant adverse effects were observed. In this randomized clinical trial of 239 early postmenopausal women, supplementation with L reuteri had no effect on bone loss or bone turnover over 2 years. The observed interaction between BMI and treatment effect warrants further investigation. ClinicalTrials.gov Identifier: NCT04169789.

Enhancing clinical decision-making: Sysmex UF-5000 as a screening tool for bacterial urinary tract infection in children.

A rapid screening test for urinary tract infections (UTIs) in children is needed to avoid unnecessary cultures and provide prompt reports to make appropriate clinical decisions. We have evaluated for the first time the performance of the Sysmex UF-5000 flow cytometer as a screening tool for UTIs in children. This study included 4445 pediatric patients, with urinary sediment and urine culture data collected from January 2020 to September 2023. The Sysmex UF-5000 analyzer was utilized to measure urine white blood cell (WBC) and bacteria (BACT), with the findings being compared to the culture results. At ≥ 104 colony-forming unit (CFU)/mL, 513 samples were culture-positive (400 samples presented 104-105 CFU/mL, and 113 demonstrated ≥ 105 CFU/mL bacterial growth). Optimal indicators for positive cultures were BACT counts of 92.2/μL (AUC: 0.944) and WBC counts of 40.8/μL (AUC:0.863). False negative rate were 0.9% when using a 7.8 bacteria/μL cut-off and avoiding unnecessary cultures in 28.1%. The UF-5000 has a higher consistency rate for Gram-negative (GN) bacteria (90.3%) than Gram-positive (GP) bacteria (86.8%). For samples with 105 CFU/mL, UF-5000's Bacteria -Information flags showed superior concordance for samples with 104-105 CFU/mL bacteria. Screening pediatric urine cultures with the UF-5000 showed potential application value in identifying negative cultures and significant bacterial growth, although performance may vary depending on the study population. Furthermore, detecting Gram typing aids in guiding early clinical empirical medication, particularly for UTIs caused by GN bacteria.

Antifungal efficacy and immunomodulatory effect of PLGA nanoparticle-encapsulated itraconazole in histoplasmosis in vivo model

IntroductionHistoplasma capsulatum is the etiological agent of histoplasmosis, the most common endemic pulmonary mycosis. Itraconazole (ITZ) is the choice for mild disease and a step-down therapy in severe and disseminated clinical presentations. Drug encapsulation into nanoparticles (NPs) is an alternative to improve drug solubility and bioavailability, reducing undesirable interactions and drug degradation and reaching the specific therapeutic target with lower doses. Objectiveevaluate the antifungal and immunomodulatory effect of ITZ encapsulated into poly(lactic-co-glycolic acid) (PLGA) NPs, administrated orally and intraperitoneally in an in vivo histoplasmosis model. ResultsAfter intranasal infection and treatment of animals with encapsulated ITZ by intraperitoneal and oral route, fungal burden control, biodistribution, immune response, and histopathology were evaluated. The results showed that the intraperitoneal administered and encapsulated ITZ has an effective antifungal effect, significantly reducing the Colony-Forming-Units (CFU) after the first doses and controlling the infection dissemination, with a higher concentration in the liver, spleen, and lung compared to the oral treatment. In addition, it produced a substantial immunomodulatory effect on pro- and anti-inflammatory cytokines and immune cell infiltrates confirmed by histopathology. ConclusionsOverall, results suggest a synergistic effect of the encapsulated drug and the immunomodulatory effect contributing to infection control, preventing their dissemination.

Bioremediation alternatives for total petroleum hydrocarbon removal in agricultural soil

Objective: The purpose of the present study is to highlight the importance of assessing bioremediation and total petroleum hydrocarbon removal by bioaugmentation and biostimulation on the rhizosphere. Design/methodology/approach: An 89-day experiment was established with treatments considering plant (corn) establishment–crude petroleum (25,000 mg kg-1)–bacteria and hydrocarbonoclastic fungi–adding nitrogen and phosphorus to agricultural soil. At the end of the experiment, hydrocarbonoclastic fungal and bacterial populations and total petroleum hydrocarbon removal were assessed. Results: Both microbial groups increased in number and time. The treatment with 120 kg nitrogen ha-1 and 12.5 kg phosphorus ha-1 allowed the highest population (227 x 103 g-1 of colony forming units (CFU) of hydrocarbonoclastic bacteria). A total of 83% petroleum hydrocarbon removal was obtained as established in 89 days. Limitations on study/implications: The effectiveness of bioremediation can vary significantly in real environments due to factors, such as soil variability, climate. Findings/conclusions: The previous results highlight the importance of using these bioremediation techniques to eliminate hydrocarbons in contaminated agricultural soils.

A syringe-driven PicoArray device for rapid isolation and quantification of foodborne Salmonella

Rapid and accurate detection of pathogenic Salmonella in food and environmental sources is of paramount importance to protect public health from salmonellosis. However, most current assays are labor-intensive, time-consuming, and require expensive instruments and experienced operators. In this work, we developed a PicoArray-based colony-forming unit (CFU) quantification system for rapid and ultrasensitive Salmonella detection. To enable on-site detection, we employed syringe-vacuum-assisted sample loading and oil-induced sample digitization to stochastically confine single bacteria in the picoliter-level wells of the PicoArray device (containing 40,233 wells). Such a single bacterial confinement enables the isolation and characterization of individual bacteria from a large population. In addition, the confinement of bacteria in small volumes allows rapid accumulation of metabolites, thereby eliminating the pre-incubation step and reducing the time required to detect bacteria. By combining it with a selective medium for the isolation of Salmonella and a fluorescent redox indicator (e.g., resazurin) to facilitate the detection of individual microcolonies, the PicoArray device can rapidly and precisely detect Salmonella. As a proof-of-concept, we demonstrated the ability of our platform for rapid, sensitive, and specific detection of Salmonella in spiked milk samples. Our platform offers the possibility of close monitoring of pathogens in food or drinks at the consumer end because it allows a non-trained operator to detect foodborne pathogens in the clinically relevant range in the home setting.

Hair removal with a clipper and microbial colonisation prior to knee arthroplasty: a randomised controlled trial

Despite the widely reported success of knee arthroplasty, studies show that 1.6-3 % of patients undergo revision within the first postoperative year predominantly due to infection. Preoperative skin preparation may potentially decrease the bacterial load and consequently, the risk of periprosthetic joint infections. The effects of hair removal on prosthetic joint infection are inconsistent. Our primary aim was to investigate if hair removal with a clipper influenced skin colonisation and bacterial composition. Forty Caucasian male participants who were planned to undergo knee arthroplasty, (mean age 63.8 years), were included. Patients were randomised to hair removal in a within-person study design. As a control, the opposite leg of the patient was used. Swabs were collected prior to hair removal (baseline), immediately after hair removal (Day 0), and with follow-up after one and seven days. The intervention showed significant decrease in mean log colony-forming units per. cm2 from baseline 2.97 to 2.67 (P<0.01) immediately after hair removal and sustained at Day 1 (P=0.01). At Day 7, the mean was non-significant compared to baseline. The control group did not show any decrease of skin microbiota at follow-up on Day 0, 1 or 7.No significant differences within the bacterial composition were found between the intervention and control leg at baseline among the six most prevalent detected bacterialspecies: Staphylococcus epidermidis, Micrococcus luteus, S. hominis, S. capitis, S. haemolyticus and S. aureus. The study did not find any changes in the bacterial composition over time. Hair removal with a clipper within 24 hours prior to surgery causes a significant non-selective reduction in skin colonisation.

Staphylococcus aureus persistence in osteocytes: weathering the storm of antibiotics and autophagy/xenophagy.

Staphylococcus aureus is a major causative pathogen of osteomyelitis. Intracellular infections of resident bone cells including osteocytes can persist despite gold-standard clinical intervention. The mechanisms by which intracellular S. aureus evades antibiotic therapy are unknown. In this study, we utilised an in vitro S. aureus infection model of human osteocytes to investigate whether antibiotic-mediated dysregulation of autophagy contributes to this phenomenon. Infected or non-infected osteocyte-like cells were exposed to combinations of rifampicin, vancomycin, and modulators of autophagy. Intracellular bacterial growth characteristics were assessed using colony-forming unit (CFU) analysis, viable bacterial DNA abundance, and the rate of escape into antibiotic-free medium, together with measures of autophagic flux. Rifampicin, alone or in combination with vancomycin, caused a rapid decrease in the culturability of intracellular bacteria, concomitant with stable or increased absolute bacterial DNA levels. Both antibiotics significantly inhibited autophagic flux. However, modulation of autophagic flux did not affect viable bacterial DNA levels. In summary, autophagy was shown to be a factor in the host-pathogen relationship in this model, as its modulation affected the growth state of intracellular S. aureus with respect to both their culturability and propensity to escape the intracellular niche. While rifampicin and vancomycin treatments moderately suppressed autophagic flux acutely, this did not explain the paradoxical response of antibiotic treatment in decreasing S. aureus culturability whilst failing to clear bacterial DNA and hence intracellular bacterial load. Thus, off-target effects of rifampicin and vancomycin on autophagic flux in osteocyte-like cells could not explain the persistent S. aureus infection in these cells.

Antimicrobial Efficacy of Saline, Chlorhexidine, and Zataria Multiflora and Mentha Piperita Essential Oils in Root Canal Irrigation of Primary Molars.

Objectives: This study aimed to compare the antimicrobial efficacy of saline, 0.5% and 2% Zataria multiflora (Z. multiflora) essential oil, 0.5% and 2% Mentha piperita (M. piperita) essential oil, and 0.2% chlorhexidine (CHX) as root canal irrigants for primary molar teeth. Materials and Methods: A total of 64 primary molars were used in this in vitro study. The teeth were randomly assigned to six groups (N=10). The root canals were prepared up to file #35, and all teeth were sterilized before contamination with Enterococcus faecalis (E. faecalis; ATCC 29212) suspension. After 48 hours of incubation, the root canals in each group were irrigated with the respective irrigants. Sterile paper points were then used to collect microbial samples from the root canals. A colony counter was used to count the number of colony-forming units (CFUs). Data were analyzed by SPSS version 20 (alpha=0.05). Results: The colony count was significantly different among the groups (P<0.001), and 2% M. piperita (P=0.009), 0.5% Z. multiflora (P=0.021), and 0.2% CHX (P=0.002) were significantly more effective than saline in elimination of E. faecalis. The ascending order of microbial count after irrigation was as follows: saline > 0.5% M. piperita > 0.2% CHX > 2% M. piperita > 0.5% Z. multiflora. Conclusion: The current study showed the optimal antibacterial activity of 0.5% Z. multiflora essential oil and 2% M. piperita essential oil against E. faecalis, and indicated their possible efficacy for use as an irrigant for root canal irrigation of primary molars.

The expression of Galectin-9 correlates with mTOR and AMPK in murine colony-forming erythroid progenitors.

Galectin-9 (Gal-9) is an immune checkpoint ligand for T-cell immunoglobulin and mucin domain 3. Although the roles of Gal-9 in regulating immune responses have been well investigated, their biological roles have yet to be fully documented. This study aimed to analyse the expression of Gal-9 bone marrow (BM) cells in C57BL/6J (B6) mice. Furthermore, the co-expression of Gal-9 with the mammalian target of rapamycin (mTOR) and AMP-activated protein kinase (AMPK) was investigated. The BM cells in adult C57BL/6J (B6) mice were collected and analysed in vitro. In a flow cytometric analysis of BM cells, Gal-9 was highly expressed in c-KithiSca-1-CD34-CD71+ erythroid progenitors (EPs), whereas it was downregulated in more differentiated c-KitloCD71+TER119+ cells. Subsequently, a negative selection of CD3-B220-Sca-1-CD34-CD41-CD16/32- EPs was performed. This resulted in substantial enrichment of KithiCD71+Gal-9+ cells and erythroid colony-forming units (CFU-Es), suggesting that the colony-forming subset of EPs are included in the KithiCD71+Gal-9+ population. Furthermore, we found that EPs had lower mTOR and AMPK expression levels in Gal-9 knockout B6 mice than in wild-type B6 mice. These results may stimulate further investigation of the role of Gal-9 in haematopoiesis.

Investigation of Freezing and Freeze-Drying for Preserving and Re-Using a Whole Microbial Cheese Community.

Preserving microbial ecosystems obtained from traditional cheese-making processes is crucial to safeguarding the biodiversity of microbial cheese communities and thus ensuring that the high flavor quality of traditional cheeses is maintained. Few protocols have been proposed for the long-term storage of microbial consortia. This work aimed to develop preservation methods to stabilize the entire microbial community in smear-ripened cheese without multiplication or isolation. A simplified microbial community, capable of reproducing the metabolic pattern of cheese maturation, was used in three independent cheese productions. Cheese samples were taken before and after the ripening step, mixed with maltodextrin or saline solution, and subjected to different stabilization conditions including freezing and freeze-drying, followed by 1 month of storage. Microbial survival was quantified using the colony-forming unit assay. Differential scanning calorimetry was used to relate the physical events occurring within the samples to the microbial storage stability. Freezing at -80 °C resulted in the lowest loss of culturability (<0.8 log unit), followed by freezing at -20 °C and freeze-drying. The ripening bacteria appeared as the most sensitive microorganisms within the community. Moreover, a successful cheese production using the best-stabilized community showed the possibility of preserving and re-using an entire microbial community of interest.

Preventive effect of prenatal maternal oral probiotic supplementation on neonatal jaundice (POPS Study): A protocol for the randomised double-blind placebo-controlled clinical trial

IntroductionNeonatal jaundice is a common and life-threatening health problem in neonates due to overaccumulation of circulating unconjugated bilirubin. Gut flora has a potential influence on bilirubin metabolism. The infant gut...

Improved automated spot counting and modeling with bias correction

ABSTRACT A complete workflow was presented for estimating the concentration of microorganisms in biological samples by automatically counting spots that represent viral plaque forming units (PFU) bacterial colony forming units (CFU), or spot forming units (SFU) in images, and modeling the counts. The workflow was designed for processing images from dilution series but can also be applied to stand-alone images. The accuracy of the methods was greatly improved by adding a newly developed bias correction method. When the spots in images are densely populated, the probability of spot overlapping increases, leading to systematic undercounting. In this paper, this undercount issue was addressed in an empirical way. The proposed empirical bias correction method utilized synthetic images with known spot sizes and counts as a training set, enabling the development of an effective bias correction function using a thin-plate spline model. Its application focused on the bias correction for the automated spot counting algorithm LoST proposed by Lin et al. Simulation results demonstrated that the empirical bias correction significantly improved spot counts, reducing bias for both fixed and random spot sizes and counts.

Injectable Platelet-Rich Fibrin and Advanced Platelet-Rich Fibrin Demonstrate Enhanced Anti-Biofilm Effect Compared to Enamel Matrix Derivatives on Decontaminated Titanium Surfaces.

The search for effective antimicrobial agents to mitigate peri-implant infections remains a crucial aspect of implant dentistry. This study aimed to evaluate and compare the antimicrobial efficacy of i-PRF, A-PRF+, and enamel matrix derivative (EMD) on decontaminated rough and smooth titanium (Ti) discs. Rough and smooth Ti discs were coated with multispecies biofilm and thoroughly debrided using a chitosan-bristled brush. Subsequently, i-PRF, A-PRF+, and EMD were applied. Untreated discs served as control. Residual adherent bacteria present on the treated Ti discs were visualized by SEM and quantified using culture technique, and colony-forming units (CFUs) were measured after 48 h and 7 days. i-PRF demonstrated better antimicrobial effectiveness on both smooth and rough implant surfaces as compared to A-PRF+ and EMD (p < 0.001). In all the experimental groups, smooth Ti discs displayed a greater reduction in microbes compared to rough Ti discs when treated with the biologics. The major reduction in CFU values was determined after seven days. i-PRF as a regenerative material may also be suitable for decontaminating implant surfaces, which could influence tissue healing and regenerative outcomes positively.