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The structural characterization and differentiation of four types of oligoubiquitin conjugates [linear (Met1)-, Lys11-, Lys48-, Lys63-linked di-, tri-, and tetraubiquitin chains] using ion mobility mass spectrometry are reported. A comparison of collision cross sections for the same linkage of di-, tri-, and tetraubiquitin chains shows differences in conformational elongation for higher charge states due to the interplay of linkage-derived structure and Coulombic repulsion. For di- and triubiquitin chains, this elongation results in a single narrow feature representing an elongated conformation type for multiple higher charge state species. In contrast, higher charge state tetraubiquitin species do not form a single conformer type as readily. A comparison of different linkages in tetraubiquitin chains reveals greater similarity in conformation type at lower charge states; with increasing charge state, the four linkage types diverge in the relative proportions of elongated conformer types with Met1- ≥ Lys11- > Lys63- > Lys48-linkage. These differences in conformational trends could be discussed with respect to biological functions of linkage-specific polyubiquitinated proteins.
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Espectrometría de Movilidad Iónica , Ubiquitina , Espectrometría de Movilidad Iónica/métodos , Ubiquitina/química , Conformación Proteica , Espectrometría de Masas/métodos , Modelos Moleculares , Lisina/químicaRESUMEN
The enhanced recovery after surgery (ERAS) protocol, including prokinetic medications, is commonly used to prevent postoperative ileus. Prospective studies evaluating the effectiveness of mosapride citrate, a prokinetic 5-hydroxytryptamine 4 receptor agonist, in patients undergoing gastrectomy within the ERAS framework are lacking. This double-blind randomized trial included patients who were scheduled for laparoscopic or robotic gastrectomy for gastric cancer. Participants were randomly assigned to either a control (placebo) or experimental (mosapride citrate) group, with drugs administered on postoperative days 1-5. Bowel motility was evaluated based on bowel transit time measured using radiopaque markers, first-flatus time, and amount of food intake. No significant differences were observed in baseline characteristics between the two groups. On postoperative day 3, no significant difference was observed in the number of radiopaque markers visible in the colon between the groups. All factors associated with bowel recovery, including the time of first flatus, length of hospital stay, amount of food intake, and severity of abdominal discomfort, were similar between the two groups. Mosapride citrate does not benefit the recovery of intestinal motility after minimally invasive gastrectomy in patients with gastric cancer. Therefore, routine postoperative use of mosapride citrate is not recommended in such patients.
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Benzamidas , Gastrectomía , Morfolinas , Neoplasias Gástricas , Humanos , Benzamidas/uso terapéutico , Flatulencia , Gastrectomía/efectos adversos , Gastrectomía/métodos , Laparoscopía/métodos , Tiempo de Internación , Morfolinas/uso terapéutico , Complicaciones Posoperatorias/prevención & control , Estudios Prospectivos , Neoplasias Gástricas/cirugía , Resultado del TratamientoRESUMEN
Intrapartum ultrasonography serves as a valuable tool for the objective evaluation of labor progression and effectively identifies underlying factors, such as asynclitism, in cases of abnormal labor progression. Among the various ultrasound measurement techniques, the angle of progression (AOP) demonstrates favorable reproducibility and accuracy in assessing fetal head descent. In the context of abnormal labor, interventions differ across different stages of labor, emphasizing the importance of investigating the utility of AOP according to labor stages in this review article. Pre-labor assessment of AOP can be beneficial in terms of counseling for the timing of induction of labor, while a wider AOP value during the prolonged first stage of labor has demonstrated a positive correlation with successful vaginal delivery and shorter time to delivery. In the second stage of labor, the AOP has exhibited efficacy in predicting the mode of delivery and complicated operative deliveries. Furthermore, it has assisted in predicting the duration of labor, thereby highlighting its potential as a decision-making model for labor progression. However, further research is needed to investigate aspects, such as the determination of cutoff values, of AOP, considering the multifaceted characteristics of labor progression, which are influenced by complex interactions among maternal, fetal, and other contributing factors.
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BACKGROUND: Iris Koreana NAKAI (IKN) is a flowering perennial plant that belongs to the Iridaceae family. In this study, we aimed to demonstrate the effects of IKN on osteoclast differentiation in vitro and in vivo. We also sought to verify the molecular mechanisms underlying its anti-osteoclastogenic effects. METHODS: Osteoclasts were formed by culturing mouse bone marrow macrophage (BMM) cells with macrophage colony-stimulating factor and receptor activator of nuclear factor-κB ligand (RANKL). Bone resorption assays were performed on dentin slices. mRNA expression levels were analyzed by quantitative polymerase chain reaction. Western blotting was performed to detect protein expression or activation. Lipopolysaccharide (LPS)-induced osteoclast formation was performed using a mouse calvarial model. RESULTS: In BMM cultures, an ethanol extract of the root part of IKN suppressed RANKL-induced osteoclast formation and bone resorptive activity. In contrast, an ethanol extract of the aerial parts of IKN had a minor effect on RANKL-induced osteoclast formation. Mechanistically, the root part of IKN suppressed RANKL-induced p38 mitogen-activated protein kinase (MAPK) activation, effectively abrogating the induction of c-Fos and nuclear factor of activated T cells 1 (NFATc1) expression. IKN administration decreased LPS-induced osteoclast formation in a calvarial osteolysis model in vivo. CONCLUSIONS: Our study suggested that the ethanol extract of the root part of IKN suppressed osteoclast differentiation and function partly by downregulating the p38 MAPK/c-Fos/NFATc1 signaling pathways. Thus, the root part.
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Ubiquitin, a conserved protein in eukaryotic cells, exists as a monomer or polyubiquitin chains known as isopeptide-linked polymers. These chains are attached to a substrate or other ubiquitin molecules through a covalent bond between the α-amino group of lysine in ubiquitin and glycine in the C-terminal of the subsequent ubiquitin unit. The choice of the specific lysine residue in ubiquitin for forming ubiquitin-ubiquitin chains determines its biochemical and biological function. A detailed chemical structure-function evaluation of the respective polyubiquitin chain is required. Interestingly, specific lysine linkage polyubiquitin chains become covalently bonded to many pathological inclusions seen in serious human disease states which appear to be resistant to normal degradation, so the interaction between polyubiquitin chains and ubiquitin antibodies is very useful. For example, the neurofibrillary tangles of Alzheimer's disease and the Lewy bodies seen in Parkinson's disease are heavily ubiquitinated and can be readily visualized using specific ubiquitin antibodies. This study utilized synthetic ubiquitin building block peptides that contained various lysine residues (K6, K11, K33, K48, and K63) linked to a Gly-Gly dipeptide, with the aim of exploring the recognition specificity of the Lys63-polyubiquitin antibody. The interaction studies between different ubiquitin building blocks and the specific Lys63-ubiquitin (K63-Ub) antibody were performed by affinity-mass spectrometry (Affinity-MS) and immunoblotting which enables direct protein identification from biological material with unprecedented selectivity. Affinity-MS and dot blot data proved the specific binding of the K63-Ub antibody to the ubiquitin peptides containing Lys6 or Lys63 residues. In epitope excision for mass spectrometric epitope identification, the ubiquitin building block with Lys63 residue bound to the immobilized K63-Ub antibody was proteolytically cleaved using pronase. The resulting epitope and non-epitope fractions were subjected to matrix-assisted laser desorption/ionization-time of flight analysis, revealing that the epitope is located within the sequence ubiquitin(60-66). Epitope extraction-MS consistently confirmed these findings.
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Poliubiquitina , Ubiquitina , Humanos , Ubiquitina/química , Ubiquitina/metabolismo , Poliubiquitina/química , Poliubiquitina/metabolismo , Lisina/química , Lisina/metabolismo , Anticuerpos , PéptidosRESUMEN
Most familial adenomatous polyposis (FAP) patients undergo total colectomy, but duodenal polyposis develops in up to 90% of patients with FAP and a 4% to 18% risk of duodenal and ampullary cancer remains. Laparoscopic pancreas-preserving near total duodenectomy is thought to be a potential option and can be an effective approach to preserve the pancreas. A 48-year-old male patient, who underwent laparoscopic total colectomy with end ileostomy because of FAP with colorectal cancer, was diagnosed with a 20 mm-sized duodenal adenoma in the second to the third portion. The operation was performed on December 27, 2021. Near total duodenectomy was done and type II Billroth gastrojejunostomy was done. Laparoscopic pancreas-sparing duodenectomy is shown to be safe, with favorable short-term oncologic outcome compared to laparoscopic pancreatoduodenectomy in terms of less blood loss, faster recovery time, and much less total cost.
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Background: Ptosis and volume atrophy of the breasts are common symptoms for various ages of women and may induce a considerable amount of stress in daily life. Periareolar augmentation mastopexy is an effective procedure for such conditions, and planning the new nipple position is very important. Aim: To provide a simple, straightforward planning and walkthrough of this operation in a journey to find the ideal level of breast lifting for natural upper fullness. Materials and methods: From January 2019 to December 2021, a total of 193 patients with volume deflation and ptosis of the breast received periareolar augmentation mastopexy in our institute. We retrospectively reviewed data on demographics, surgical procedures, outcomes, and complications. Results: All operations were done with periareolar incisions, and the mean follow-up period was 29.48 ± 9.11 months. The Likert scale of outcome satisfaction scored 9.02 ± 0.61. Complications were minimal, and no symmastia or bottoming out occurred. Conclusion: We present our basic strategies of periareolar augmentation mastopexy with a slight modification of the design. We believe that lifting the nipple to 3-4 cm above the inframammary fold (IMF) (making the top of the areola about 5-6 cm above the IMF) yields satisfactory aesthetic results.
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Li-free all-solid-state batteries can achieve high energy density and safety. However, separation of the current collector/solid electrolyte interface during Li deposition increases interfacial resistance, which deteriorates safety and reversibility. In this study, a reversible 3D porous anode is designed based on Li deposition behavior that depends on the pore size of the anode. More Li deposits are accommodated within the smaller pores of the Li hosting anode composed of Ni particles with a granular piling structure; this implies the Li movement into the anode is achieved via diffusional Coble creep. Surface modification of Ni with a carbon coating layer and Ag nanoparticles further increases the Li hosting capacity and enables Li deposition without anode/solid electrolyte interface separation. A Li-free all-solid-state full cell with a LiNi0.8 Mn0.1 Co0.1 O2 cathode shows an areal capacity of 2 mAh cm-2 for retaining a Coulombic efficiency of 99.46% for 100 cycles at 30 °C.
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Transduodenal ampullectomy (TDA) is the treatment of choice for large premalignant lesions of the ampulla of Vater (AoV). With the development of surgical techniques, various methods, including the open, laparoscopic, and robotic approaches, for performing TDA have emerged. Herein, we report four consecutive cases treated with open, laparoscopic, and robotic TDA, with technical pitfalls and future perspectives of TDA in treating premalignant lesions of the AoV. The surgical techniques and principles for TDA were the same regardless of the surgical approaches. After surgery, none of the patients showed any abnormal findings or complications, except for digestive problems. All these surgical approaches are appropriate for patients requiring TDA; however, minimally invasive TDA, particularly the robotic approach is ideal. Considering the surgical complexity of TDA, the robotic approach is considered to be effective.
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A recently developed treatment strategy for lung cancer that combines immune checkpoint inhibitors with chemotherapy has been applied as a standard treatment for lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC), and it has improved the outcomes of chemotherapy. Maintenance treatment with anti-PD-1 antibody (aPD-1) enhances the effect of immunochemical combination therapy and improves therapeutic efficacy, which contributes toward a significant improvement in patient survival rates. The AXL receptor tyrosine kinase (AXL), which is expressed in tumor cells, plays an essential role in the resistance of cancers to chemotherapy and immunotherapy, and stimulates signaling associated with epithelial-mesenchymal transition (EMT) in metastatic cancer. AXL is thus an attractive target for controlling resistance to anti-tumor therapies. In this study, we examined the effect of AXL inhibitors on immune activation and tumor growth in TC1 and C3PQ mouse tumor models, in the context of clinical immunotherapy/chemotherapy and maintenance treatment, using an aPD-1 with/without pemetrexed. To determine the optimal timing for administration of SKI-G-801, an AXL inhibitor, we investigated its anti-tumor effects based on inclusion at the immunochemotherapy and maintenance therapy stages. We also performed flow cytometry-based immune profiling of myeloid cells and lymphoid cells at different points in the treatment schedule, to investigate the immune activation and anti-tumor effects of the AXL inhibitor. The addition of SKI-G-801 to the immune checkpoint inhibitor and chemotherapy stage, as well as the maintenance therapy stage, produced the best anti-tumor results, and significant tumor growth inhibition was observed in both the TC1 and C3PQ models. Both models also exhibited increased proportion of effector memory helper T cells and increased expression of CD86+ macrophages. Especially, regulatory T cells were significantly reduced in the TC1 tumor model and there was an increase in central memory cytotoxic T cell infiltration and an increased proportion of macrophages with high CD80 expression in the C3PQ tumor model. These results suggest increased infiltration of T cells, consistent with previous studies using AXL inhibitors. It is expected that the results from this study will serve as a stepping stone for clinical research to improve the existing standard of care.
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As the demand for rechargeable lithium-ion batteries (LIBs) with higher energy density increases, the interest in lithium-rich oxide (LRO) with extraordinarily high capacities is surging. The capacity of LRO cathodes exceeds that of conventional layered oxides. This has been attributed to the redox contribution from both cations and anions, either sequentially or simultaneously. However, LROs with notable anion redox suffer from capacity loss and voltage decay during cycling. Therefore, a fundamental understanding of their electrochemical behaviors and related structural evolution is a prerequisite for the successful development of high-capacity LRO cathodes with anion redox activity. However, there is still controversy over their electrochemical behavior and principles of operation. In addition, complicated redox mechanisms and the lack of sufficient analytical tools render the basic study difficult. In this review, we aim to introduce theoretical insights into the anion redox mechanism andin situanalytical instruments that can be used to prove the mechanism and behavior of cathodes with anion redox activity. We summarized the anion redox phenomenon, suggested mechanisms, and discussed the history of development for anion redox in cathode materials of LIBs. Finally, we review the recent progress in identification of reaction mechanisms in LROs and validation of engineering strategies to improve cathode performance based on anion redox through various analytical tools, particularly,in situcharacterization techniques. Because unexpected phenomena may occur during cycling, it is crucial to study the kinetic properties of materialsin situunder operating conditions, especially for this newly investigated anion redox phenomenon. This review provides a comprehensive perspective on the future direction of studies on materials with anion redox activity.
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In this paper, we present a new regularized image reconstruction method for positron emission tomography (PET), where an adaptive weighted median regularizer is used in the context of a penalized-likelihood framework. The motivation of our work is to overcome the limitation of the conventional median regularizer, which has proven useful for tomographic reconstruction but suffers from the negative effect of removing fine details in the underlying image when the edges occupy less than half of the window elements. The crux of our method is inspired by the well-known non-local means denoising approach, which exploits the measure of similarity between the image patches for weighted smoothing. However, our method is different from the non-local means denoising approach in that the similarity measure between the patches is used for the median weights rather than for the smoothing weights. As the median weights, in this case, are spatially variant, they provide adaptive median regularization achieving high-quality reconstructions. The experimental results indicate that our similarity-driven median regularization method not only improves the reconstruction accuracy, but also has great potential for super-resolution reconstruction for PET.
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Algoritmos , Procesamiento de Imagen Asistido por Computador , Procesamiento de Imagen Asistido por Computador/métodos , Fantasmas de Imagen , Tomografía de Emisión de Positrones/métodos , Tomografía Computarizada por Rayos XRESUMEN
Non-coding microRNAs (miRNAs) have been proposed to play diverse roles in cancer biology, including epithelial-mesenchymal transition (EMT) crucial for cancer progression. Previous comparative studies revealed distinct expression profiles of miRNAs relevant to tumorigenesis and progression of oral cancer. With putative targets of these miRNAs mostly validated in vitro, it remains unclear whether similar miRNA-target relationships exist in vivo. In this study, we employed a hybrid approach, utilizing both Drosophila melanogaster and human oral cancer cells, to validate projected miRNA-target relationships relevant to EMT. Notably, overexpression of dme-miR-133 resulted in significant tissue growth in Drosophila larval wing discs. The RT-PCR analysis successfully validated a subset of its putative targets, including Pde1c. Subsequent experiments performed in oral cancer cells confirmed conserved targeting of human PDE1C by hsa-miR-133. Furthermore, the elevated level of miR-133 and its targeting of PDE1C was positively correlated with enhanced migrative ability of oral cancer cells treated with LPS, along with the molecular signature of a facilitated EMT process induced by LPS and TGF-ß. The analysis on the RNAseq data also revealed a negative correlation between the expression level of hsa-miR-133 and the survival of oral cancer patients. Taken together, our mammal-to-Drosophila-to-mammal approach successfully validates targeting of PDE1C by miR-133 both in vivo and in vitro, underlying the promoted EMT phenotypes and potentially influencing the prognosis of oral cancer patients. This hybrid approach will further aid to widen our scope in investigation of intractable human malignancies, including oral cancer.
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BACKGROUD/OBJECTIVES: Alzheimer's disease (AD) is the most common cause of dementia in the elderly. Due to the increased incidence of dementia, there is a corresponding increase concerning the importance of AD. In this study, we investigated the protective effects conferred by Zizyphus jujuba (Zj) and Zizyphus jujuba fermented by yeast (Zj-Y), on cognitive impairment in an AD mouse model. MATERIALS/METHODS: AD was induced by injecting amyloid beta25-35 (Aß25-35) in ICR mice, and subsequently 200 mg/kg Zj or Zj-Y was administered daily for 14 days. The cognitive ability of AD mice was observed through behavioral experiments in T-maze, novel object recognition, and Morris water maze tests. We subsequently measured the levels of malondialdehyde (MDA), nitric oxide (NO), aspartate aminotransferase, and alanine aminotransferase in either tissues or serum. RESULTS: In behavioral tests, deterioration was revealed in the short- and long-term learning and memory functions in the Aß25-35-injected control group compared to the normal group, indicating that Aß25-35 injection impairs cognitive functions. However, administration of Zj and Zj-Y improved cognitive function in mice, as compared to the Aß25-35-injected control mice. In addition, the Aß25-35 induced elevations of MDA and NO in the brain, kidney, and liver were suppressed after exposure to Zj and Zj-Y. Especially, Zj-Y showed stronger scavenging effect against MDA and NO, as compared to Zj. CONCLUSIONS: Results of the present study indicate that Zj-Y exerts a protective effect on cognitive impairment and memory dysfunction, which is exerted by attenuating the oxidative stress induced by Aß25-35.
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Recent comparative studies have indicated distinct expression profiles of short, non-coding microRNAs (miRNAs) in various types of cancer, including oral squamous cell carcinoma (OSCC). In this study, we employed a hybrid approach using Drosophila melanogaster as well as OSCC cell lines to validate putative targets of oral cancer-related miRNAs both in vivo and in vitro. Following overexpression of Drosophila miR-31, we found a significant decrease in the size of the imaginal wing discs and downregulation of a subset of putative targets, including wntless (wls), an important regulator of the Wnt signaling pathway. Parallel experiments performed in OSCC cells have also confirmed a similar miR-31-dependent regulation of human WLS that was not initially predicted as targets of human miR-31. Furthermore, we found subsequent downregulation of cyclin D1 and c-MYC, two of the main transcriptional targets of Wnt signaling, suggesting a potential role of miR-31 in regulating the cell cycle and proliferation of OSCC cells. Taken together, our Drosophila-based in vivo system in conjunction with the human in vitro platform will thus provide a novel insight into a mammal-to-Drosophila-to-mammal approach to validate putative targets of human miRNA and to better understand the miRNA-target relationships that play an important role in the pathophysiology of oral cancer.
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Proteínas de Drosophila/genética , Péptidos y Proteínas de Señalización Intracelular/genética , MicroARNs/genética , Neoplasias de la Boca/genética , Receptores Acoplados a Proteínas G/genética , Carcinoma de Células Escamosas de Cabeza y Cuello/genética , Animales , Apoptosis/genética , Línea Celular Tumoral , Proliferación Celular/genética , Drosophila melanogaster/genética , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Neoplasias de la Boca/patología , Carcinoma de Células Escamosas de Cabeza y Cuello/patología , Vía de Señalización Wnt/genéticaRESUMEN
We recently implemented highly sensitive detection systems for photo-sensitizing potassium ions (K+) based on two-step Förster resonance energy transfer (FRET). As a successive study for quantitative understanding of energy transfer processes in terms of the exciton population, we investigated the fluorescence decay dynamics in conjugated polymers and an aptamer-based 6-carboxyfluorescein (6-FAM)/6-carboxytetramethylrhodamine (TAMRA) complex. In the presence of K+ ions, the Guanine-rich aptamer enabled efficient two-step resonance energy transfer from conjugated polymers to dyed pairs of 6-FAM and TAMRA through the G-quadruplex phase. Although the fluorescence decay time of TAMRA barely changed, the fluorescence intensity was significantly increased. We also found that 6-FAM showed a decreased exciton population due the compensation of energy transfer to TAMRA by FRET from conjugated polymers, but a fluorescence quenching also occurred concomitantly. Consequently, the fluorescence intensity of TAMRA showed a 4-fold enhancement, where the initial transfer efficiency (~300%) rapidly saturated within ~0.5 ns and the plateau of transfer efficiency (~230%) remained afterward.
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This study was conducted to understand alcohol kinetics for Koreans and to determine whether an individual is in absorption phase or elimination phase at the time of blood collection by analyzing of ethyl glucuronide and ethyl sulfate in blood. A total of 50 healthy adults was selected and assigned to drink 1g of ethanol per kg body weight of individual within 1h. Blood samples were then collected every 15min for the first 3h, 30min next 3h, and 1h last 9h. Urine samples were also collected from the individual, but not under the controlled environment. All samples were then analyzed by gas chromatography with a flame ionization detector (GC-FID) for alcohol and liquid chromatography-mass/mass spectrometry (LC-MS/MS) for EtG and EtS. The maximum BAC (Cmax) was 0.138% (g/100mL) in average under the controlled experimental condition. Alcohol elimination rates (ß) in average were 0.020% for male and 0.024% for female, respectively. It was found that the ratio of UAC and BAC was less than 1 in the absorption phase and the average ratio of UAC and BAC was 1.47 in the elimination phase. The comparison of BAC (g/L) and EtG (mg/L) absorption and elimination curves showed that the intersection time was 3.9h in average. It is shown that the ratio of EtG (mg/L)/BAC (g/L) is higher than 1, the individual would be in elimination phase of BAC. At the time of Cmax, the ratio of EtG (mg/L)/BAC (g/L) was 0.255±0.132 (SD) in average.
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Consumo de Bebidas Alcohólicas/sangre , Depresores del Sistema Nervioso Central/farmacocinética , Etanol/farmacocinética , Glucuronatos/sangre , Ésteres del Ácido Sulfúrico/sangre , Adulto , Pueblo Asiatico , Biomarcadores/sangre , Depresores del Sistema Nervioso Central/sangre , Depresores del Sistema Nervioso Central/orina , Cromatografía de Gases , Cromatografía Liquida , Etanol/sangre , Etanol/orina , Femenino , Humanos , Masculino , Espectrometría de Masas , Persona de Mediana Edad , República de Corea , Adulto JovenRESUMEN
The relationship between sugar level and development of dental caries has long been a main topic in dentistry. However, as a ubiquitous component of the modern diet, sucrose is mainly derived from three meals a day, rather than a long time exposure. In this study, various concentrations of sucrose were provided to Streptococcus mutans biofilms for 1 h per exposure (three times per day) to imitate a human meal pattern. And then the relationship between sucrose concentration and changes in the treated biofilms was determined. The results indicated that the components and acid production of the treated biofilms changed in a second-order polynomial curve pattern with sucrose concentration increase, which were confirmed by CLSM and SEM analyses. However, gene expression related to extracellular polysaccharides (EPS) formation, acid production and tolerance was up-regulated with sucrose concentration increase, which might have been due to compensation for the decrease in EPS formation and acid production by the biofilms at higher concentrations of sucrose. These findings suggest that sucrose in the range of 1%-5% can support the highest acid production and accumulation of S. mutans biofilms, which may further increase its cariogenic potential. However, additional studies are required to confirm the relationships in human cariogenic biofilms.
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Biopelículas/crecimiento & desarrollo , Caries Dental/patología , Polisacáridos Bacterianos/biosíntesis , Streptococcus mutans/metabolismo , Sacarosa/metabolismo , Ácidos/metabolismo , Caries Dental/microbiología , Humanos , Microscopía Confocal , Microscopía Electrónica de RastreoRESUMEN
The aim of this study was to investigate the difference in composition and virulence of Streptococcus mutans biofilms according to substratum direction. S. mutans biofilms (46-h-old) were formed on three different saliva-coated hydroxyapatite (sHA) disc direction groups: downward (discs placed in the direction of gravity), vertical (discs placed parallel to gravity direction), and upward (discs placed opposite to gravity). The 46-h-old biofilms on sHA discs in the upward direction showed the highest biofilm accumulation, colony forming unit (CFU) count, and extracellular polysaccharide (EPS) amount, followed by those in the vertical and downward directions. In the confocal laser scanning microscopy (CLSM) study, the biofilms in the upward direction also showed the highest bacterial count (live or dead cells) and EPS biovolume. Scanning electron microscopy (SEM) analysis confirmed the microbiological and biochemical results. In addition, biofilm density and acid production were higher in the upward direction than those in the other directions. Our findings suggest that substratum direction, which might be related to gravity, strongly influences the formation and virulence of cariogenic biofilms and subsequent initiation of dental caries. Collectively, the differences in the formation and virulence of cariogenic biofilms are related to the direction of tooth surface (occlusal surfaces of mandibular teeth > proximal surfaces > occlusal surfaces of maxillary teeth).
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Biopelículas/crecimiento & desarrollo , Caries Dental/microbiología , Virulencia , Durapatita/química , Microscopía Electrónica de Rastreo , Streptococcus mutans , Propiedades de SuperficieRESUMEN
The present study investigated the effect of periodic 1-min fluoride treatments on Streptococcus mutans biofilms and then determined the relationship between anti-biofilm activity, treatment frequency, and fluoride concentration using a linear-fitting procedure. S. mutans biofilms were periodically treated (1-min/treatment) with fluoride during biofilm formation and analyzed using microbiological methods, confocal microscopy, and real-time PCR. The results indicated that reductions in the dry weight and acidogenicity of biofilms due to periodic fluoride treatment occurred in a concentration dependent manner. The reduction in dry weight without affecting bacterial cell viability was observed mainly due to the inhibitory effect of fluoride on gtfB and gtfC gene expression, which suppresses EPS production and avoids reduction of the pH below the critical point on the tooth surface. This study suggests that brief periodic exposure to appropriate fluoride concentrations through mouthwashes and toothpastes may affect the virulence and composition of cariogenic biofilms and subsequently prevent dental caries.