• IMMUNE NETWORK
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• v.20 no.3
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• pp.25.1-25.13
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• 2020

Acinetobacter baumannii is known for its multidrug antibiotic resistance. New approaches to treating drug-resistant bacterial infections are urgently required. Cathelicidin-related antimicrobial peptide (CRAMP) is a murine antimicrobial peptide that exerts diverse immune functions, including both direct bacterial cell killing and immunomodulatory effects. In this study, we sought to identify the role of CRAMP in the host immune response to multidrug-resistant Acinetobacter baumannii. Wild-type (WT) and CRAMP knockout mice were infected intranasally with the bacteria. CRAMP^-/- mice exhibited increased bacterial colony-forming units (CFUs) in bronchoalveolar lavage (BAL) fluid after A. baumannii infection compared to WT mice. The loss of CRAMP expression resulted in a significant decrease in the recruitment of immune cells, primarily neutrophils. The levels of IL-6 and CXCL1 were lower, whereas the levels of IL-10 were significantly higher in the BAL fluid of CRAMP^-/- mice compared to WT mice 1 day after infection. In an in vitro assay using thioglycollate-induced peritoneal neutrophils, the ability of bacterial phagocytosis and killing was impaired in CRAMP^-/- neutrophils compared to the WT cells. CRAMP was also essential for the production of cytokines and chemokines in response to A. baumannii in neutrophils. In addition, the A. baumannii-induced inhibitor of κB-α degradation and phosphorylation of p38 MAPK were impaired in CRAMP^-/- neutrophils, whereas ERK and JNK phosphorylation was upregulated. Our results indicate that CRAMP plays an important role in the host defense against pulmonary infection with A. baumannii by promoting the antibacterial activity of neutrophils and regulating the innate immune responses.

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.154-161
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• 1998

To investigate whether the introduced genetic marker is useful to detect the survivalship and activity of the natural bacteria under the stressed conditions, one Gram-negative isolate, KP964 was transformed to the luminous phenotype by transferring luxAB gene. Under the starvation-stress this luminous bacterial culturability (determined by colony-forming-units [CFU] on agar plate) decreased rapidly below the detection limit by 37 days, while its total cell number (determined by AODC) remained almost the same as its initial inocular size. At that time period, the viable cell number was estimated to be 1400 times higher than its CFU number. The bioiuminescence (determined by relative light units [RLU]) produced under the same condition was also monitored and found to decrease more rapidly than the culturability by 5-fold. Under the other stresses, e.g., osmotic shocks, acid shock, and exposure to toxic chemicals, this bacterial strain did not show the reliable correlation between CFU and RLU. These results might not suggest the direct estimation of bioiuminescence from the stressed bacteria be an index of both the survivalship and its activity. However, when the stressed bacterial cells were incubated under the favorable condition by relieving from the existing stress, the potential bioiuminescence (the lag periods before the increase of bioiuminescence, the increase rates of bioiuminescence, and the maximal levels of bioiuminescence) was shown to be highly dependent upon the strengths of the stresses exposed to the bacterial cells. Therefore, analysis of the potential bioiuminescence from the stressed bacteria revealed good relationships with survival as well as activity.

• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1401-1407
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• 2009

Many Lactobacillus strains have been promoted as good probiotics for the prevention and treatment of diseases. We engineered recombinant Lactobacillus casei, producing biologically active canine granulocyte macrophage colony stimulating factor (cGM-CSF), and investigated its possibility as a good probiotic agent for dogs. Expression of the cGM-CSF protein in the recombinant Lactobacillus was confirmed by SDS-PAGE and Western blotting methods. For the in vivo study, 18 Beagle puppies of 7 weeks of age were divided into three groups; the control group was fed only on a regular diet and the two treatment groups were fed on a diet supplemented with either $1\times10^9$ colony forming units (CFU)/day of L. casei or L. casei expressing cGM-CSF protein for 7 weeks. Body weight was measured, and fecal and blood samples were collected from the dogs during the experiment for the measurement of hematology, fecal immunoglobulin (Ig)A and IgG, circulating IgA and IgG, and canine corona virus (CCV)-specific IgG. There were no differences in body weights among the groups, but monocyte counts in hematology and serum IgA were higher in the group receiving L. casei expressing cGM-CSF than in the other two groups. After the administration of CCV vaccine, CCV-specific IgG in serum increased more in the group supplemented with L. casei expressing cGM-CSF than the other two groups. This study shows that a dietary L. casei expressing cGM-CSF enhances specific immune functions at both the mucosal and systemic levels in puppies.

• Animal Bioscience
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• v.37 no.8
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• pp.1418-1427
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• 2024

Objective: This study aimed to investigate the efficacy of Bacillus-based probiotics supplemented at two different levels to modulate the productive performance, egg quality, tibia traits, and specific cecal bacteria counts of Hy-Line Brown layers from 25 to 37 weeks of age. Methods: A total of 216 twenty-five-week-old hens were randomly distributed into 3 experimental diets with 12 replicates of 6 birds per cage. Diets included basal diet supplemented with 0 (CON), 3×10⁸ (PRO1), or 3×10⁹ (PRO2) colony-forming unit (CFU) of the test probiotic containing Bacillus subtilis PB6, Bacillus subtilis FXA, and Bacillus licheniformis G3 per kilogram of feed. Results: Improved egg weights and mass at 29 weeks; and feed intake at 31 weeks (p<0.10) were noticed with the probiotic-supplemented PRO1 and PRO2 diets. Considering egg quality, the shell thickness, Haugh units, and yolk color were improved; but yolk cholesterol was lowered (p<0.05) with PRO1 and PRO2 diets at 29 weeks. At both 33 and 37 weeks, the egg-breaking strength, shell color and thickness, albumen height, Haugh units, and yolk color were improved; but yolk cholesterol was similarly lowered (p<0.05) with the PRO1 and PRO2 diets. Improved tibia Ca, ash, weights, and density; and raised cecal counts of Bifidobacteria and Lactobacilli (p<0.05) were noticed with PRO1 and PRO2 diets. Improved tibia P but reduced Clostridia counts (p<0.10) were also observed with the PRO1 and PRO2 diets. Conclusion: Probiotic supplementation of Bacillus subtilis PB6, Bacillus subtilis FXA, and Bacillus licheniformis G3 at 3×10⁸ CFU/kg of feed is adequate to significantly improve egg quality, lower yolk cholesterol, enhance several tibia traits, and raise the populations of beneficial cecal bacteria. Modest improvements in several productive parameters and tibia P but reduced Clostridia were also observed; and could warrant further investigation of probiotic effects beyond the current test period.

• Clinical and Experimental Pediatrics
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• v.48 no.9
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• pp.986-990
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• 2005

Purpose : Certain strains of lactobacilli are known to accelerate recovery from acute diarrhea. Lactobacillus reuteri is isolated from human breast milk and a commonly occurring Lactobacillus species with therapeutic potential in acute diarrhea. The purpose of the present study was to investigate the therapeutic effect of L. reuteri in acute diarrhea in young children. Methods : Fifty patients between 6 and 36 months of age hospitalized with acute diarrhea (rotavirus in 40 percent) were randomized into two groups to receive either $10^8$ colony-forming units of L. reuteri or a matching placebo, twice a day for their length of hospitalization, or for up to 5 days. Antidiarrheal drugs were not prescribed to either group. The clinical outcome of diarrhea was evaluated. Results : The mean duration of watery diarrhea after initiation of treatment was 2.3 days for the L. leuteri group(n=25) vs. 2.9 days for the placebo group(n=25)(P=0.072). By the second day of treatment, watery diarrhea persisted in 64 percent of patients receiving L. reuteri, compared to 84 percent of those receiving placebo(P=0.006). On the second day, the mean frequency of watery diarrhea was 1.9 in the L. leuteri group and 3.4 in the placebo(P=0.046). Also, vomiting continued to the second day in 16 percent of patients receiving L. reuteri and 40 percent of those recieving placebo(P=0.031). Conclusion : L. reuteri is effective as a therapeutic agent in acute diarrhea in children.

• Tuberculosis and Respiratory Diseases
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• v.55 no.1
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• pp.59-68
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• 2003

Background : In intubated patients, cultures of endotracheal aspirates (EA) are apt to contamination throughout the endotracheal tube. Therefore, the identification of etiologic agents via conventional EA cultures is not always reliable. In order to differentiate a pulmonary infection from a non-infectious disease, and to identify the true etiologic agent of acute pulmonary infection, blinded protected specimen brushing (PSB) was used, and its efficacy evaluated. Methods : In 51 intubated patients, with suspected pneumonia, blind PSB were performed, and the results compared with blood and EA cultures. A protected specimen brush was introduced through the endotracheal tube, and settled at the affected large bronchus. A specimen brush was introduced to the expected region using the blind method. The tip of the brush was introduced with an aseptic technique after vigorously mixed for 1 minute in $1cm^3$ of Ringer's lactate solution. The specimens were submitted for quantitative culture within 15 minutes, with a culture being regarded as positive if the colony forming units were above $10^3/ml$. Results : Of the 51 patients, 15 (29.4%) had community-acquired pneumonia (CAP), 27 (52.9%) hospital-acquired pneumonia (HAP) and 9 (17.6%) non-infectious diseases. The sensitivity and specificity of the quantitative PSB culture for the diagnosis of pneumonia were 52.4 and 88.9%, respectively. The sensitivity and specificity of EA were 78.6 and 77.8%, respectively. The blind PSB was superior to the EA for the identification of true etiologic agents. Of 53 episodes of 27 HAP patients, MRSA (Methicillin-resistant staphylococcus aureus) (41.5%) was the most common causative agent followed by Pseudomonas aeruginosa (15.1%), Klebsiella sp. (7.5%) and Acinetobacter sp. (7.5%). Conclusions : As a simple, non-invasive diagnostic modality, the blind PSB is a useful method for the differentiation of a pulmonary infection from non-infectious diseases and to identify the etiologic agents in intubated patients. A blind PSB can be performed without bronchoscopy, so is safer, more convenient and cost-effectiveness for patients where bronchoscopy can not be performed.

• Journal of Dental Rehabilitation and Applied Science
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• v.40 no.2
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• pp.55-63
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• 2024

Purpose: This study aimed to assess the antimicrobial efficacy of an 810-nm infrared diode laser with indocyanine green (ICG) against Staphylococcus aureus on sandblasted, large grit, and acid-etched (SLA) titanium surfaces, comparing its effectiveness with alternative chemical decontamination modalities. Materials and Methods: Biofilms of S. aureus ATCC 25923 were cultured on SLA titanium disks for 48 hours. The biofilms were divided into five treatment groups: control, chlorhexidine gluconate (CHX), tetracycline (TC), ICG, and 810-nm infrared diode laser with ICG (ICG-PDT). After treatment, colony-forming units were quantified to assess surviving bacteria, and viability was confirmed through confocal laser-scanning microscope (CLSM) imaging. Results: All treated groups exhibited a statistically significant reduction in S. aureus (P < 0.05), with notable efficacy in the CHX, TC, and ICG-PDT groups (P < 0.01). While no statistical difference was observed between TC and CHX, the ICG-PDT group demonstrated superior bacterial reduction. CLSM images revealed a higher proportion of dead bacteria stained in red within the ICG-PDT groups. Conclusion: Within the limitations, ICG-PDT effectively reduced S. aureus biofilms on SLA titanium surfaces. Further investigations into alternative decontamination methods and the clinical impact of ICG-PDT on peri-implant diseases are warranted.

• Restorative Dentistry and Endodontics
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• v.42 no.4
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• pp.316-323
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• 2017

Objectives: This study compared the amount of apically extruded bacteria during the glide-path preparation by using multi-file and single-file glide-path establishing nickel-titanium (NiTi) rotary systems. Materials and Methods: Sixty mandibular first molar teeth were used to prepare the test apparatus. They were decoronated, blocked into glass vials, sterilized in ethylene oxide gas, infected with a pure culture of Enterococcus faecalis, randomly assigned to 5 experimental groups, and then prepared using manual stainless-steel files (group KF) and glide-path establishing NiTi rotary files (group PF with PathFiles, group GF with G-Files, group PG with ProGlider, and group OG with One G). At the end of canal preparation, 0.01 mL NaCl solution was taken from the experimental vials. The suspension was plated on brain heart infusion agar and colonies of bacteria were counted, and the results were given as number of colony-forming units (CFU). Results: The manual instrumentation technique tested in group KF extruded the highest number of bacteria compared to the other 4 groups (p < 0.05). The 4 groups using rotary glide-path establishing instruments extruded similar amounts of bacteria. Conclusions: All glide-path establishment instrument systems tested caused a measurable apical extrusion of bacteria. The manual glide-path preparation showed the highest number of bacteria extruded compared to the other NiTi glide-path establishing instruments.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.9
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• pp.1340-1349
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• 2017

Objective: This study aimed to evaluate the microbiological and cellular milk profile for the diagnosis of subclinical mastitis in female buffaloes and to assess risk factors for predisposition of the disease. Methods: Analyses were carried out by standard plate count (SPC), identification of species and antibiotic resistance, somatic cell count (SCC), electrical electrical conductivity of milk (ECM), and lactoferrin content in milk. Teat cups were swabbed to evaluate risk factors, observing hyperkeratosis, milking vacuum pressure and cleanliness of the site. Hence, 30 female buffaloes were randomly selected (15 from a group in early lactation and 15 in late lactation). Results: The most common bacteria in the microbiological examination were Staphylococcus spp., Streptococcus spp. and Corynebacterium sp. In the antibiotic sensitivity test, 10 (58.82%) of the 17 antibiotics tested were sensitive to all isolates, and resistant bacteria were Streptococcus uberis, Streptococcus dysgalactiae, Streptococcus haemolyticus, and Escherichia coli. It was observed that positive samples in the microbiological examination showed total bacterial count between $9.10{\times}10^3$ to $6.94{\times}10^6$ colony forming units/mL, SCC between 42,000 to 4,320,000 cells/mL and ECM ranging from 1.85 to 7.40 mS/cm. It was also found that the teat cups had high microbial counts indicating poor hygiene, and even faults in the cleanliness of the animals' waiting room were observed. It is concluded that values of SCC above 537,000 cells/mL and ECM above 3.0 mS/mL are indications of mammary gland infection for this herd; however, the association of these values with a microbiological analysis is necessary to more accurately evaluate the health status of mammary glands with subclinical mastitis. Conclusion: Through phenotypic characterization of bacteria involved in the samples, the genera Staphylococcus spp., Streptococcus spp., and Corynebacterimum bovis were the most prevalent in this study. Faults in environment and equipment hygienization are factors that are directly associated with mastitis.

• Journal of Periodontal and Implant Science
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• v.39 no.3
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• pp.303-310
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• 2009

Purpose: The aerosol generated by ultrasonic scaler can contain bacteria or virus which can penetrate into body through respiratory systems of dentists, dental hygienist or patients. The aim of this study is to evaluate the effect of chlorhexidine digluconate as preoperative mouthrinse or lavage for ultrasonic scaler on the reduction of viable organisms in aerosol produced during periodontal treatment using ultrasonic scaler. Methods: 30 patients with moderate chronic periodontitis were included and divided into 3 groups: Control (no preoperative mouthrinse and tap water as lavage), CHG (preoperative mouthrinse with 0.1% chlorhexidine digluconate and tap water as lavage), CHL (no preoperative mouthrinse and 0.1% chlorhexidine digluconate as lavage). Each patient received scaling or subgingival curettage for 30 min. In CHG group, mouthrinse with chlorhexidine digluconate was performed for 1 min. before treatment. Before, during and after scaling or subgingival curettage, air sampling was performed for 7 min. each (1000 L/7 min.) with trypticase-soy agar plate. Agar plates were incubated in $37^{\circ}C$ aerobically. The numbers of colony-forming units (CFU) were counted and compared. Results: The numbers of CFUs of the samples obtained during treatment were $97{\pm}14.0$ in control, $73.1{\pm}14.9$ in CHG group and $44.5{\pm}9.0$ in CHL group. The difference among the 3 groups was determined to be statistically significant (one-way ANOVA with Bonferroni's correction, p-value: 0.0003). In contrast, the numbers of CFU of samples obtained before and after treatment were not significantly different among the groups. Conclusions: Chlorhexidine digluconate used as preoperative mouthrinse or lavage for ultrasonic scaler can reduce the microorganisms in aerosol produced during periodontal treatment using ultrasonic scaler. Less number of microorganisms were detected when chlorhexidine was used as lavage for ultrasonic scaler.