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Prevalence and Molecular Characterization of Methicillin-Resistant Staphylococcus aureus from Nasal Specimens: Overcoming MRSA with Silver Nanoparticles and Their Applications

  • Aly E. Abo-Amer;Sanaa M. F. Gad El-Rab;Eman M. Halawani;Ameen M. Niaz;Mohammed S. Bamaga
    • Journal of Microbiology and Biotechnology
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    • v.32 no.12
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    • pp.1537-1546
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    • 2022
  • Staphylococcus aureus is a cause of high mortality in humans and therefore it is necessary to prevent its transmission and reduce infections. Our goals in this research were to investigate the frequency of methicillin-resistant S. aureus (MRSA) in Taif, Saudi Arabia, and assess the relationship between the phenotypic antimicrobial sensitivity patterns and the genes responsible for resistance. In addition, we examined the antimicrobial efficiency and application of silver nanoparticles (AgNPs) against MRSA isolates. Seventy-two nasal swabs were taken from patients; MRSA was cultivated on Mannitol Salt Agar supplemented with methicillin, and 16S rRNA sequencing was conducted in addition to morphological and biochemical identification. Specific resistance genes such as ermAC, aacA-aphD, tetKM, vatABC and mecA were PCR-amplified and resistance plasmids were also investigated. The MRSA incidence was ~49 % among the 72 S. aureus isolates and all MRSA strains were resistant to oxacillin, penicillin, and cefoxitin. However, vancomycin, linezolid, teicoplanin, mupirocin, and rifampicin were effective against 100% of MRSA strains. About 61% of MRSA strains exhibited multidrug resistance and were resistant to 3-12 antimicrobial medications (MDR). Methicillin resistance gene mecA was presented in all MDR-MRSA strains. Most MDR-MRSA contained a plasmid of > 10 kb. To overcome bacterial resistance, AgNPs were applied and displayed high antimicrobial activity and synergistic effect with penicillin. Our findings may help establish programs to control bacterial spread in communities as AgNPs appeared to exert a synergistic effect with penicillin to control bacterial resistance.

Studies on the Isolation, Culture and Fusion of Protoplasts from Plant Mesophyll and Cells Cultured in vitro (식물원형질체의 분리, 배양 및 융합에 관한 연구)

  • Choi, S.J.;Son, S.H.;Chang, W.C.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.27 no.2
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    • pp.147-154
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    • 1982
  • This study was conducted to investigate an effective method of protoplast isolation, the plating efficiency for cell division, and fusion of plant protoplasts by polyethylene glycol for somatic hybridzation. The effectiveness of protoplast isolation was different with the various enzyme concentrations, but, in the protoplast isolation from tobacco mesophyll, the enzyme solution with 0.5% macerozyme and 2.0% cellulase was very effective. The protoplast isolation from callus cultured in vitro for a long period was not obtained in any of the enzyme solution used. Protoplasts divided actively at cell densities above $10^44/ml and at $25^{\circ}C$ under 12hr illumination by inflorecient light (l50 Lux), regardless of presence of agar. The highest frequency of protoplast fusion was obtained after treatment with a solution of 0.33 M polyethylene glycol 1500.

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Clinical Outcomes Associated with Degree of Hypernatremia in Neurocritically Ill Patients

  • Yun Im, Lee;Joonghyun, Ahn;Jeong-Am, Ryu
    • Journal of Korean Neurosurgical Society
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    • v.66 no.1
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    • pp.95-104
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    • 2023
  • Objective : Hypernatremia is a common complication encountered during the treatment of neurocritically ill patients. However, it is unclear whether clinical outcomes correlate with the severity of hypernatremia in such patients. Therefore, we investigated the impact of hypernatremia on mortality of these patients, depending on the degree of hypernatremia. Methods : Among neurosurgical patients admitted to the intensive care unit (ICU) in a tertiary hospital from January 2013 to December 2019, patients who were hospitalized in the ICU for more than 5 days and whose serum sodium levels were obtained during ICU admission were included. Hypernatremia was defined as the highest serum sodium level exceeding 150 mEq/L observed. We classified the patients into four subgroups according to the severity of hypernatremia and performed propensity score matching analysis. Results : Among 1146 patients, 353 patients (30.8%) showed hypernatremia. Based on propensity score matching, 290 pairs were included in the analysis. The hypernatremia group had higher rates of in-hospital mortality and 28-day mortality in both overall and matched population (both p<0.001 and p=0.001, respectively). In multivariable analysis of propensity score-matched population, moderate and severe hypernatremia were significantly associated with in-hospital mortality (adjusted odds ratio [OR], 4.58; 95% confidence interval [CI], 2.15-9.75 and adjusted OR, 6.93; 95% CI, 3.46-13.90, respectively) and 28-day mortality (adjusted OR, 3.51; 95% CI, 1.54-7.98 and adjusted OR, 10.60; 95% CI, 5.10-21.90, respectively) compared with the absence of hypernatremia. However, clinical outcomes, including in-hospital mortality and 28-day mortality, were not significantly different between the group without hypernatremia and the group with mild hypernatremia (p=0.720 and p=0.690, respectively). The mortality rates of patients with moderate and severe hypernatremia were significantly higher in both overall and matched population. Interestingly, the mild hypernatremia group of matched population showed the best survival rate. Conclusion : Moderate and severe hypernatremia were associated with poor clinical outcomes in neurocritically ill patients. However, the prognosis of patients with mild hypernatremia was similar with that of patients without hypernatremia. Therefore, mild hypernatremia may be allowed during treatment of intracranial hypertension using hyperosmolar therapy.

Comparison of the metabolic profile of the mycelia and fruiting bodies of artificially cultured Cordyceps militaris

  • Ha, Si Young;Jung, Ji Young;Park, Han Min;Yang, Jae-Kyung
    • Journal of Mushroom
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    • v.20 no.1
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    • pp.13-21
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    • 2022
  • Cordyceps militaris, a well-known traditional Chinese medicine, has multiple health-promoting effects. It is used as a herbal remedy and health food in Asian countries. Cultured mycelia are often used as a substitute for natural C. militaris. In the present study, the mycelia and fruiting bodies of artificially cultured C. militaris were analyzed using a metabolomics approach. The protein and crude fat contents of the mycelia were substantially higher than those of the fruiting bodies. The top three abundant amino acids in the mycelia were proline (3.9 g/100 g), aspartic acid (2.9 g/100 g), and glutamic acid (2.7 g/100 g). The carbohydrate content was similar in the fruiting bodies and mycelia. Analysis revealed that both the fruiting bodies and mycelia are rich in phenolic compounds and exhibit antioxidant activity. Further, six metabolites were significantly different between the mycelia and fruiting bodies. The levels of Ca, glucose, Mg, and Se were higher in the mycelia than in the fruiting bodies. In contrast, mannitol and Zn were more abundant in the fruiting bodies. The current study provides a comprehensive metabolic profile of the mycelia and fruiting bodies of artificially cultured C. militaris. Such an exercise is potentially important for understanding the metabolism of C. militaris and facilitating the use of cultured mycelia as a supplement to C. militaris fruiting bodies in traditional Chinese medicine.

Effect of black chokeberry on skeletal muscle damage and neuronal cell death

  • Kim, Jisu;Lee, Kang Pa;Beak, Suji;Kang, Hye Ra;Kim, Yong Kyun;Lim, Kiwon
    • Korean Journal of Exercise Nutrition
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    • v.23 no.4
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    • pp.26-31
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    • 2019
  • [Purpose] Numerous epidemiological studies have shown that it is possible to prescribe exercise for neurodegenerative disease, such as Alzheimer's disease and Parkinson's disease. However, despite the availability of diverse scientific knowledge, the effects of exercise in this regard are still unclear. Therefore, this study attempted to investigate a substance, such as black chokeberry (Aronia melanocapa L.) that could improve the ability of the treatment and enhance the benefits of exercising in neurodegenerative diseases. [Methods] The cell viability was tested with 2,3-bis[2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolim-5-carboxanilide and the cells were stained with ethidium homodimer-1 solution. The mRNA expression levels were evaluated by microarray. The active compounds of black chokeberry ethanolic extract (BCE) were analyzed by gas chromatography. The chemical shift analysis in the brain was performed using magnetic resonance spectroscopy. [Results] BCE treatment decreased hydrogen peroxide-induced L6 cell death and beta amyloid induced primary neuronal cell death. Furthermore, BCE treatment significantly reduced the mRNA levels of the inflammatory factors, such as IL-1α, Cxcl13, IL36rn, Itgb2, Epha2, Slamf8, Itgb6, Kdm6b, Acvr1, Cd6, Adora3, Cd27, Gata3, Tnfrsf25, Cd40lg, Clec10a, and Slc11a1, in the primary neuronal cells. Next, we identified 16 active compounds from BCE, including D-mannitol. In vivo, BCE (administered orally at a dosage of 50 mg/kg) significantly regulated chemical shift in the brain. [Conclusion] Our findings suggest that BCE can serve as a candidate for neurodegenerative disease therapy owing to its cyto-protective and anti-inflammatory effects. Therefore, BCE treatment is expected to prevent damage to the muscles and neurons of the athletes who continue high intensity exercise. In future studies, it would be necessary to elucidate the effects of combined BCE intake and exercise.

Production of Exopolysaccharides and İndole Acetic Acid (IAA) by Rhizobacteria and Their Potential against Drought Stress in Upland Rice

  • Tetty Marta Linda;Jusinta Aliska;Nita Feronika;Ineiga Melisa;Erwina Juliantari
    • Journal of Microbiology and Biotechnology
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    • v.34 no.6
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    • pp.1239-1248
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    • 2024
  • Peatlands are marginal agricultural lands due to highly acidic soil conditions and poor drainage systems. Drought stress is a big problem in peatlands as it can affect plants through poor root development, so technological innovations are needed to increase the productivity and sustainability of upland rice on peatlands. Rhizobacteria can overcome the effects of drought stress by altering root morphology, regulating stress-responsive genes, and producing exopolysaccharides and indole acetic acid (IAA). This study aimed to determine the ability of rhizobacteria in upland rice to produce exopolysaccharides and IAA, identify potential isolates using molecular markers, and prove the effect of rhizobacteria on viability and vigor index in upland rice. Rhizobacterial isolates were grown on yeast extract mannitol broth (YEMB) medium for exopolysaccharides production testing and Nutrient Broth (NB)+L-tryptophan medium for IAA production testing. The selected isolates identify using sequence 16S rRNA. The variables observed in testing the effect of rhizobacteria were germination ability, vigour index, and growth uniformity. EPS-1 isolate is the best production of exopolysaccharides (41.6 mg/ml) and IAA (60.83 ppm). The isolate EPS-1 was identified as Klebsiella variicola using 16S rRNA sequencing and phylogenetic analysis. The isolate EPS-1 can increase the viability and vigor of upland rice seeds. K. variicola is more adaptive and has several functional properties that can be developed as a potential bioagent or biofertilizer to improve soil nutrition, moisture and enhance plant growth. The use of rhizobacteria can reduce dependence on the use of synthetic materials with sustainable agriculture.

In vitro Development Potential Following Nuclear Transfer of Porcine Interspecies Clone Embryo by Goat Somatic Cells (유산양 체세포를 이용한 돼지 난자의 이종간 핵이식 후 배발달에 관한 연구)

  • Chang, Suk-Min;Naruse, Kenji;Shin, Young-Min;Park, Chang-Sik;Jin, Dong-Il
    • Korean Journal of Agricultural Science
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    • v.33 no.1
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    • pp.35-41
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    • 2006
  • This study was conducted to investigate the developmental ability of interspecies cloned embryos after nuclear transfer of goat fetal fibroblast cells into porcien oocytes. Recipient porcine and goat oocytes were obtained from slaughterhouse and matured in vitro according to established protocols. Enucleation was accomplished by aspirating the first polar body and cytoplasm and a single donor cell was individually microinjected into vitelline space of the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3M mannitol fusion medium. After electro-fusion, interspecies reconstituted embryos were cultured in PZM-3 for 7 days. In porcine interspecies nuclear transfer with goat fetal fibroblast cells, the cleavage rate of reconstituted embryos were 58.9% which was no significant different from that in porcine nuclear transfer embryos (67.4%). However, the developmental rate into blastocyst stage was 5.4% in interspecies nuclear transfer which was significantly lower than that in porcine intraspecies nuclear transfer (13.6%). When the developmental ability of porcine interspecies nuclear transfer with goat cells was compared with goat intraspecies nuclear transfer, the cleavage rate of embryos were 59.2% and the developmental rate into morular and blastocyst stage was 13.6% in interspecies nuclear transfer which were significantly lower than those in intraspecies nuclear transfer embryos. This result indicated that porcine interspecies nuclear transfer with goat fetal fibroblast cells showed the developmental potential in vitro with lower cleavage and developmental rate compared with intraspecies nuclear transfer.

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Comparison of Boiling Point and Distillaiion Ranige, Melting Range, and Identification Methods of Various Organizations on Synthetic Food Additives (식품첨가물에 대한 여러 기관의 비점 및 유분측정법, 융점측정법 및 확인시험법 비교)

  • Shin Dong-Hwa;Kim Yong-Suk;Lee Young-Hwan;Bang Jeong-Ho;Om Ae-Son;Shin Jae-Wook;Lee Tal-Soo;Jang Young-Mi;Hong Ki-Hyoung;Park Sung-Kwan;Kwon Yong-Kwan;Park Jae-Seok
    • Journal of Food Hygiene and Safety
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    • v.20 no.3
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    • pp.134-140
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    • 2005
  • Boiling point and distillation range, melting range, and identification methods in general test method of Korea, Japan, Joint FAO/WHO Expert Committee of Food Additives (JECFA), and USA on chemical food additives were compared. Boiling point of propylene glycol was indicated as boiling point in Korea, distillate in Japan, distillation range in JECFA and USA, and its value was up to the standard. Distillation range of propionic acid was indicated as distillate in Korea and Japan, distillation range in JECFA and USA, and its value was up to the standard. There is no standard on distillation range of isopropyl alcohol in Japanese method. Test method of melting range on synthetic food additives was identical in all organizations, and there are 28 items to which this test method applies in Korean Food Additives Code. The standards on molting range of D-mannitol were different in various organizations, and in USA method there are no standards to which L-ascorbic acid, calciferol, and fumaric acid apply. Synthetic food additives performing the identification test were 251 items in Korean Food Additives Code, but there are no items to which manganese, glycerophosphate, bromate, thiosulfate, and bromide apply. Calcium benzoate was dissolved by heating in benzoate test and we could not identify the citrate in ferric citrate by method (2) of Korea and Japan. Identification test methods for ammonium, lactate, magnesium, copper, sulfate, phosphate, and zinc were identical in all organizations, and these could be identifed by current identification methods.

Mathematical Models to Predict Staphylococcus aureus Growth on Processed Cheeses

  • Kim, Kyungmi;Lee, Heeyoung;Moon, Jinsan;Kim, Youngjo;Heo, Eunjeong;Park, Hyunjung;Yoon, Yohan
    • Journal of Food Hygiene and Safety
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    • v.28 no.3
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    • pp.217-221
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    • 2013
  • This study developed predictive models for the kinetic behavior of Staphylococcus aureus on processed cheeses. Mozzarella slice cheese and cheddar slice cheese were inoculated with 0.1 ml of a S. aureus strain mixture (ATCC13565, ATCC14458, ATCC23235, ATCC27664, and NCCP10826). The inoculated samples were then stored at $4^{\circ}C$ (1440 h), $15^{\circ}C$ (288 h), $25^{\circ}C$ (72 h), and $30^{\circ}C$ (48 h), and the growth of all bacteria and of S. aureus were enumerated on tryptic soy agar and mannitol salt agar, respectively. The Baranyi model was fitted to the growth data of S. aureus to calculate growth rate (${\mu}_{max}$; ${\log}CFU{\cdot}g^{-1}{\cdot}h^{-1}$), lag phase duration (LPD; h), lower asymptote (log CFU/g), and upper asymptote (log CFU/g). The growth parameters were further analyzed using the square root model as a function of temperature. The model performance was validated with observed data, and the root mean square error (RMSE) was calculated. At $4^{\circ}C$, S. aureus cell growth was not observed on either processed cheese, but S. aureus growth on the mozzarella and cheddar cheeses was observed at $15^{\circ}C$, $25^{\circ}C$, and $30^{\circ}C$. The ${\mu}_{max}$ values increased, but LPD values decreased as storage temperature increased. In addition, the developed models showed acceptable performance (RMSE = 0.3500-0.5344). This result indicates that the developed kinetic model should be useful in describing the growth pattern of S. aureus in processed cheeses.

Biological Activities and Cultural Characteristics of an Entomogenous fungus, Paecilomyces tenuipes (Peck) Samson (눈꽃동충하초의 배양적 특성 및 생리활성)

  • Ha, Nam-Gyu;Kim, Seung-Yul;Kang, Jin-Ho;Kang, Pil-Don;Sung, Gyoo-Byung;Hong, In-Pyo
    • Journal of Sericultural and Entomological Science
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    • v.47 no.1
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    • pp.12-17
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    • 2005
  • To develop technique for the production of P. tenuipes stromata on a large scale, the infection of P. tenuipes and the growth of stroma were investigated by silkworm (Bombyx mori) variety. Also, studied about biological activites of fruiting body formed on silkworm. Infection rate of the 5th instar larvae of the silkworm with P. tenuipes was the highest in Yangwonjam, followed by Hachojam, Baegokjam and Chilbojam in that order. Also, as the inoculation times was increased, infection rate tended to be raised. The rate of fruiting body formation of the silkworm pupae infected with P. tenuipes was the highest in Baegokjam, followed by Yangwonjam and Chilbojam in the order. But, actually the fruiting body formation of the 5th instar larvae of the silkworm tested was good in Chilbojam, followed by Yangwonjam and Baegokjam in that order in 3 times spraying inoculation. The fruiting bodies of Yangwonjam and Chilbojam infected with P. tenuipes had high amount of Mannitol, but Baegokjam and Hachojam had high concentration of Glucose on a dry weight basis. The mean content of total amino acid in the fruiting bodies of P. tenuipes was 1.03 ${\mu}mole/g$. The distribution rate of amino acid components decreased in the order of Arginine (12.2%)>Glycine (10.5%)> Proline (9.6)>Tyrosine (8.9%)>Serine>Leucine>Threonine. The most abundant amino acid in the fruiting bodies of the Baegokjam, Chilbojam and Hachojam infected with P. tenuipes was arginine, while Yangwonjam was Glycine. The most abundant fatty acid in P.tenuipes was Oleic acid on a dry weight basis. The unsaturated fatty acids such as Oleic acid, Linoleic acid and Linolenic acid accounted for more than 78% of the total fatty acids.