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Production of Poly (3-Hydroxybutyrate-co-3-Hydroxyvalerate) by Bacillus sp. EMK-5020 Using Makgeolli Lees Enzymatic Hydrolysate and Propionic Acid as Carbon Sources (막걸리 주박 가수분해 산물과 propionic acid를 탄소원으로 이용한 Bacillus sp. EML-5020 균주로부터 poly (3-hydroxybutyrate-co-3-hydroxyvalerate) 생합성)

  • Kwon, Kyungjin;Kim, Jong-Sik;Chung, Chung-Wook
    • Journal of Life Science
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    • v.32 no.7
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    • pp.510-522
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    • 2022
  • In this study, to biosynthesize PHA with properties more similar to polypropylene, a Bacillus sp. EMK-5020 strain that biosynthesized poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) was isolated from soil. Bacillus sp. EMK-5020 strain biosynthesized PHBV containing 1.3% 3-hydroxyvalerate (3HV) using reducing sugar contained in Makgeolli lees enzymatic hydrolysate (MLEH) as a single carbon source. As the amount of propionic acid, which was added as a second carbon source, increased, the content of 3HV also increased. PHBV containing up to 48.6% of 3HV was synthesized when 1.0 g/l of propionic acid was added. Based on these results, the strain was cultured for 72 hr in a 3 l fermenter using reducing sugar in MLEH (20 g/l) and propionic acid (1 g/l) as the main and secondary carbon sources, respectively. As a result, 6.4 g/l DCW and 50 wt% of PHBV (MLEH-PHBV) containing 8.9% 3HV were biosynthesized. Through gel permeation chromatography and thermogravimetric analysis, it was confirmed that the average molecular weight and the decomposition temperature of MLEH-PHBV were 152 kDa and 273℃, respectively. In conclusion, the Bacillus sp. EMK-5020 strain could biosynthesize PHBV containing various 3HV fractions when MLEH and propionic acid were used as carbon sources, and PHBV-MLEH containing 8.9% 3HV was confirmed to have higher thermal stability than standard PHBV (8% 3HV).

Grapevine Growth and Berry Development under the Agrivoltaic Solar Panels in the Vineyards (영농형 태양광 시설 설치에 따른 포도나무 생육 및 과실 특성 변화 비교)

  • Ahn, Soon Young;Lee, Dan Bi;Lee, Hae In;Myint, Zar Le;Min, Sang Yoon;Kim, Bo Myung;Oh, Wook;Jung, Jae Hak;Yun, Hae Keun
    • Journal of Bio-Environment Control
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    • v.31 no.4
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    • pp.356-365
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    • 2022
  • Agrivoltaic systems, also called solar sharing, stated from an idea that utilizes sunlight above the light saturation point of crops for power generation using solar panels. The agrivoltaic systems are expected to reduce the incident solar radiation, the consequent surface cooling effect, and evapotranspiration, and bring additional income to farms through solar power generation by combining crops with solar photovoltaics. In this study, to evaluate if agrivoltaic systems are suitable for viticulture, we investigated the microclimatic change, the growth of vines and the characteristics of grape grown under solar panels set by planting lines compared with ones in open vineyards. There was high reduction of wind speed during over-wintering season, and low soil temperature under solar panel compared to those in the open field. There was not significant difference in total carbohydrates and bud burst in bearing mother branches between plots. Despite high content of chlorophyll in vines grown under panels, there is no significant difference in shoot growth of vines, berry weight, cluster weight, total soluble solid content and acidity of berries, and anthocyanin content of berry skins in harvested grapes in vineyards under panels and open vineyards. It was observed that harvesting season was delayed by 7-10 days due to late skin coloration in grapes grown in vineyards under panels compared to ones grown in open vineyards. The results from this study would be used as data required in development of viticulture system under panel in the future and further study for evaluating the influence of agrivoltaic system on production of crops including grapes.

The Protective Role of Gleditsiae fructus against Streptococcus pneumoniae (폐렴 구균에 대한 조협의 보호 역할 연구)

  • Jun-ki Lee;Se-Hui Lee;Dong Ju Seo;Kang-Hee Lee;Sojung Park;Sun Park;Taekyung Kim;Jin-Young Yang
    • Journal of Life Science
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    • v.33 no.2
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    • pp.158-168
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    • 2023
  • Natural products have been used to mitigate the effects of cancer and infectious diseases, as they feature diverse bioactivities, such as antioxidant, antibacterial, anti-inflammatory, and immunomodulatory effects. Here, we chose 10 natural products that are well-known as pulmonary enhancers and investigated their bactericidal effects on Streptococcus pneumoniae. In the disk diffusion assay, the growth of S. pneumoniae was significantly regulated by G. fructus treatment regardless of extraction method used. We first adopted spraying as a novel delivery method for G. fructus. Interestingly, mice exposed to G. fructus three times a day for 2 weeks were resistant to S. pneumoniae intranasal infection (shown both through body weight loss and survival rates compared to the control group). Moreover, we confirmed that exposure to G. fructus regulated the colonization of the bacteria despite the sustained inflammation in the lung after exposure to S. pneumoniae, indicating that migrated inflammatory immune cells may involve a host defense mechanism against pulmonary infectious diseases. While a similar number of granulocytes (CD11b+Ly6C+Ly6G+), neutrophils (CD11b+Ly6CintLy6G+), and monocytes (CD11b+Ly6CintLy6G-) were found between groups, a significantly increased number of alveolar macrophages (CD11b+CD11chiF4/80+) was detected in BAL fluids of mice pre-exposed to G. fructus at 5 days after S. pneumonia infection. Taken together, our data suggest that this usage of G. fructus can induce protective immunity against bacterial infection, indicating that facial spray may be helpful in enhancing the defense mechanism against pulmonary inflammation and in evaluating the efficacy of natural products as immune enhancers against respiratory diseases.

Preselection of Bovine Blastocysts Expressing Exogeneous Gene Following Microinjection (외래유전자를 주입한 소 수정란에서 형질전환가능 수정란의 선발)

  • 공일근
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.167-176
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    • 1997
  • This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

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Studies on the Changes of Reproductive Organs, Serum Sex Hormones and Metabolites according to the Gestation Period in Rabbit (가토(家兎)의 임신(姙娠)에 따른 생식기관(生殖器官), 혈중(血中) 성(性)Hormone 및 대사물질(代謝物質)의 변화(變化)에 관(關)한 연구(硏究))

  • Lee, Kyu Seung;Han, Sung Wook;Park, Chang Sik
    • Korean Journal of Agricultural Science
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    • v.9 no.1
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    • pp.303-313
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    • 1982
  • The study was conducted to find out the concentrations of sex hormones and the contents of metabolites in serum, and the changes of weights and tissues on ovary, thyroid gland and adrenal gland according to gestation period in rabbit. The results were summarized as follows: 1. The ovary weights were increased significantly with the time e lapse after gestation and recovered normally at 5 days after parturition. In the histological changes of ovary, the lutein cells were hypertrophied and the secretory granules were increased actively until 3 weeks after gestation, and then a trophied thereafter. 2. The thyroid gland weights at all observation times were higher than those in control group, and the significance was recognized at 1, 3 and 4 weeks after gestation. The histological features of the secretory epithelium were the hyper trophic and columnar condition stimulating the functional state from 1 week after gestation. 3. The adrenal gland weights in experimental group were recognized significantly at 4 weeks after gestation, but showed higher than those in control group at all observation times. The zona fasciculata and zona reticularis of the gland showed the slight hypertrophic condition, but the zona glomeerulosa and adrenal medulla did not find out any particular changes. 4. The serum concentrations of progesterone and LH reached a peak level at 2 weeks and 1 week after geestation respectively, and rapidly began to decline thereafter. 5. The serum concentrations of estradiol-$17{\beta}$ and FSH were not detected below 20.0 pg/ml and 1.3 mIU/ml respectively. 6. The contents of total protein and non-protein nitrogen nitrogen were decreased gradually with the time elapse after gestation, but the significant differences were recognized from 3 weeks. 7. The total lipids were not changed markedly until 3 weeks, but increased significantly at 4 weeks after gestation and at 5 days after parturition. 8. The serum cholesterol tended to be decreased until 3 weeks, but increased at 4 weeks after gestation and at 5 days after parturition. 9. The serum calcium showed a continuous decrease during the gestation period, but the significant differences were recognized at 3 and 4 weeks. The serum phosphorus also had a significant decrease at 4 weeks after gestation.

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Peripheral Neutrophil Count and Respiratory Failure in Preterm Infant (조산아에서 말초혈액 중성구수와 호흡 부전증과의 연관성)

  • Lee, Kum Joo;Yun, Soo Young;Lee, Ran;Hean, Jae Ho;Jung, Ghee Young;Park, Jin Hee;Park, Young Sun
    • Clinical and Experimental Pediatrics
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    • v.45 no.5
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    • pp.596-602
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    • 2002
  • Purpose : The purpose of this study was to analyze the association of peripheral neutrophil count with the development of respiratory failure in preterm infants. Methods : A retrospective study was conducted from January 1993 to December 1999 on 44 preterm infants, who were admitted to the neonatal intensive care unit of St. Francisco hospital. Preterm infants(birth weight 500 to 1,350 gm) who had a complete blood count obtained within 2 hours after delivery. Patients in the lowest of neutrophil count(early neutropenia, < $1.0{\times}10^9/L$) were compared with patients in the remaining group. Results : Low neutrophil count were transient in early neutropenia group. The concentration the circulating neutrophil count rose from $0.85{\pm}0.11{\times}10^9/L$ at average of 2 hours after delivery to $5.3{\pm}2.7{\times}10^9/L$ at 24 hours after delivery in the early neutropenia group and from $3.6{\pm}1.6{\times}10^9/L$ to $5.8{\pm}3.2{\times}10^9/L$ in the non-neutropenia group during the same time period. Compare to the non-neutropenia group, the neutropenia group had a lower birth weight($1,046.50{\pm}180.76gm$ Vs $1,156.70{\pm}124.99gm$), a lower Apgar score(1 min : $3.41{\pm}1.18$ Vs $4.30{\pm}1.46$, 5 min : $5.41{\pm}0.87$ Vs $6.15{\pm}0.95$), and a higher incidence of bronchopulmonary dysplasia(27.27% Vs 7.0%). Patients who had early neutropenia were more likely to require mechanical ventilation, supplemental oxygen and hospital stay. Also, main effect factors for the two groups were birth weight(Odds ratio=5.457, 95% CI=1.551-27.525), initial peripheral blood white cells(odds ratio=8.308, 95% CI=2.054-52.699), and bronchopulmonary dysplasia(odds ratio=0.099, 95% CI=0.017-0.397). Conclusion : A low count of neutrophil in the systemic circulation of premature infants within 2 hours of birth is associated with more severe respiratory distress.

Pulmonary Mycoses in Immunocompromised Hosts (면역기능저하 환자에서 폐진균증에 대한 임상적 고찰)

  • Suh, Gee-Young;Park, Sang-Joon;Kang, Kyeong-Woo;Koh, Young-Min;Kim, Tae-Sung;Chung, Man-Pyo;Kim, Ho-Joong;Han, Jong-Ho;Choi, Dong-Chull;Song, Jae-Hoon;Kwon, O-Jung;Rhee, Chong-H.
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.6
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    • pp.1199-1213
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    • 1998
  • Background : The number of immunocompromised hosts has been increasing steadily and a new pulmonary infiltrate in these patients is a potentially lethal condition which needs rapid diagnosis and treatment. In this study we sought to examine the clinical manifestations, radiologic findings, and therapeutic outcomes of pulmonary mycoses presenting as a new pulmonary infiltrate in immunocompromised hosts. Method : All cases presenting as a new pulmonary infiltrate in immunocompromised hosts and confirmed to be pulmonary mycoses by pathologic examination or by positive culture from a sterile site between October of 1996 and April of 1998 were included in the study and their chart and radiologic findings were retrospectively reviewed. Results : In all, 14 cases of pulmonary mycoses from 13 patients(male : female ratio = 8 : 5, median age 47 yr) were found. Twelve cases were diagnosed as aspergillosis while two were diagnosed as mucormycosis. Major risk factors for fungal infections were chemotherapy for hematologic malignancy(10 cases) and organ transplant recipients(4 cases). Three cases were receiving empirical amphotericin B at the time of appearance of new lung infiltrates. Cases in the hematologic malignancy group had more prominent symptoms : fever(9/10), cough(6/10), sputum(5/10), dyspnea(4/10), chest pain(5/10). Patients in the organ transplant group had minimal symptoms(p<0.05). On simple chest films, all of the cases presented as single or multiple nodules(6/14) or consolidations(8/14). High resolution computed tomograph showed peri-lesional ground glass opacities(14/14), pleural effusions(5/14), and cavitary changes(7/14). Definitive diagnostic methods were as follows : 10 cases underwent minithoracotomy, 2 underwent video-assisted thoracoscopic surgery, 1 underwent percutaneous needle aspiration and 1 case was diagnosed by culture of abscess fluid. All cases received treatment with amphotericin B with 1 case each being treated with liposomal amphotericin B and itraconazole due to renal toxicity. Lung lesion improved in 12 of 14 patient but 4 patients died before completing therapy. Conclusion : When a new lung infiltrate develops presenting either as a nodule or consolidation in a neutropenic patient with hematologic malignancy or in a transplant recipient, you should always consider pulmonary mycoses as one of the differential diagnosis. By performing aggressive work up and early treatment, we may improve prognosis of these patients.

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STUDIES ON AVIAN VISCERAL LYMPHOMATOSIS I. THE INCREASED INCIDENSE AMONG CHICKEN FLOCKS AND PATHOLOGIC PICTURES (장기형임파종증(臟器型淋巴腫症)에 관(關)한 연구(硏究) 1. 계군(鷄群)에서의 임파종증(淋巴腫症)의 발생(發生) 및 병리학적소견(病理學的所見))

  • Kim, Uh Ho;Lim, Chang Hyeong
    • Korean Journal of Veterinary Research
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    • v.4 no.1
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    • pp.35-42
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    • 1964
  • 1). An nanlysis was made of 3,500 postmortem diagnoses for the three years 1961 through 1963 to determine whether there was any actual incidence of avian visceral lymphomntosis in the field. Chickens autopsied, which showed gross alterations were 7.6 percent or 266 cases. The diminished incidence of the disease in second and third years seemed due to decreased total numbers of chicken flocks year by year for the reason of difficult feed supply. 2). Because chickens autopsied in this study were not clearly known of their breeds and lines, no distinct data on the incidence in various breeds were made. Some exact breeds were in too small numbers to have any statistical significance. Inconceivably, no other types of avian leukosis than visceral lymphomatosis had been observed in any appreciable number in this analysis. 3). Pathologic analysis for affected organs was made grossly and microscopically. In the gross pictures, liver, spleen, kidney, ovary, and in some case, intestine principally showed lesions, but its manifestation was variable in different organs. In such organs, livers were affected more frequently, and spleens followed next. The organs were classified and arranged according to the gross alterations, and among their distribution one-half of livers were in diffuse variety; one-fourths in nodular; about one-sevenths in mixed; and granular variety followed next. In the spleen samples, two-thirds were in diffuse variety; one-fourths in nodular; and follicular only in three cases. Ovaries almost showed follicular lesions, the diffused were less than one-fifths of total specimens. Kidneys were occurred almost in diffuse variety. And intestine showed only nodular tomors. Microscopically, 42 cases of visceral lymphomatosis composed of 24 livers, 10 spleens, 3 kidneys, 3 intestines and 2 ovaries were examined. The tumor cells were lymphoid cells showing various component in size, shape and stainability. Mitotic figures were usually present. The proportion of the component cells were various in all cases and there were variations in the distribution of the tumor cells. The types of distribution were classified according to the standard proposed by Horiuchi as nodular, infiltrative and diffuse proliferation. In cases of visceral lymphomatosis of the livers and the spleens the types of infiltrative, nodular and diffuse proliferation could be classified. In the cases of the kidneys the types of diffuse and nodular proliferation were observed. In the cases of the intestines and the ovaries the types of infiltrative and diffuse proliferation were observed respectively.

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Multiplication of Infectious Flacherie and Densonucleosis Viruses in the Silkworm, Bombyx mori (가잠의 전염성 연화병 및 농핵병 바이러스 증식에 관한 연구)

  • 김근영;강석권
    • Journal of Sericultural and Entomological Science
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    • v.25 no.2
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    • pp.1-31
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    • 1984
  • Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.

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