• Proceedings of the Korean Society for Applied Microbiology Conference
• /
• 2001.06a
• /
• pp.141-145
• /
• 2001

Isopentenyl diphosphate (IPP) is the common, five-carbon building block in the biosynthesis of all carotenoids. IPP in Escherichia coli is synthesized through the non-mevalonate pathway. The first reaction of IPP biosynthesis in E. coli is the formation of l-deoxy-D-xylulose-5-phosphate (DXP), catalyzed by DXP synthase and encoded by dxs. The second reaction in the pathway is the reduction of DXP to 2-C-methyl-D-erythritol-4-phosphate, catalyzed by DXP reductoisomerase and encoded by dxr. To determine if one or more of the reactions in the non-mevalonate pathway controlled flux to IPP, dxs and dxr were placed on several expression vectors under the control of three different promoters and transformed into three E. coli strains (DH5$\alpha$, XL1-Blue, and JMl0l) that had been engineered to produce lycopene. Lycopene production was improved significantly in strains transformed with the dxs expression vectors. When the dxs gene was expressed from the arabinose-inducible araBAD promoter ( $P_{BAD}$) on a medium-copy plasmid, lycopene production was 2-fold higher than when dxs was expressed from the IPTG-inducible trc and lac promoters ( $P_{trc}$ and $P_{lac}$, respectively) on medium-copy and high-copy plasmids. Given the low final densities of cells expressing dxs from IPTG-inducible promoters, the low lycopene production was probably due to the metabolic burden of plasmid maintenance and an excessive drain of central metabolic intermediates. At arabinose concentrations between 0 and 1.33 roM, cells expressing both dxs and dxr from $P_{BAD}$ on a medium-copy plasmid produced 1.4 - 2.0 times more lycopene than cells expressing dxs only. However, at higher arabinose concentrations lycopene . production in cells expressing both dxs and dxr was lower than in cells expressing dxs only. A comparison of the three E. coli strains transformed with the arabinose-inducible dxs on a medium-copy plasmid revealed that lycopene production was highest in XLI-Blue.LI-Blue.

• Natural Product Sciences
• /
• v.26 no.1
• /
• pp.64-70
• /
• 2020

Citrus junos seeds (CS) have been traditionally used for the treatment of cancer and neuralgia. They are also used to manufacture edible oil and cosmetic perfume. A large amount of CS shells without oil (CSS) are discarded after the oil in CS is used as foods or herbal remedy. To efficiently utilize CSS as a by-products, it needs to be studied through chemical analysis. Therefore, we developed an ultra-performance liquid chromatography (UPLC)-diode array detection (DAD) method for simultaneous determination and quantitative analysis of five components (two flavonoids and threes limonoids) in CSS. A Waters Acquity UPLC HSS T3 column C18 (2.1 × 100 mm, 1.8 ㎛) was used for this separation. It was maintained at 40 ℃. The mobile phase used for the analysis was distilled water and acetonitrile with gradient elution. To identify the quantity of the five components, a mass spectrometer (MS) with an electrospray ionization (ESI) source was used. The regression equation showed great linearity, with correlation coefficient ≥ 0.9912. Limits of detection (LOD) and limits of quantification (LOQ) of the five compounds were 0.09 - 0.13 and 0.26 - 0.38 ㎍/mL, respectively. Recoveries of extraction ranged from 97.45% to 101.91%. Relative standard deviation (RSD) values of intra- and inter-day precision were 0.06 - 1.15% and 0.19 - 0.25%, respectively. This UPLC-DAD method can be validated to simultaneously analyze quantities of marker flavonoids and limonoids in CSS.

• Journal of Applied Biological Chemistry
• /
• v.63 no.1
• /
• pp.69-74
• /
• 2020

The fruit, leaves, and roots of Elaeagnus multiflora Thunb. have been used in traditional Chinese medicine to treat cough, diarrhea, and itching. However, the anti-fatigue effects of E. multiflora fruit (EMF) extract have not been studied in detail. The aim of this study was to examine the effect of EMF on fatigue and exercise performance in BALB/c mice. EMF was orally administered to mice at four doses (10, 50, 100, and 200 mg/kg/day) for 2 weeks. The anti-fatigue activity was evaluated by determining the exhaustive swimming time. Blood lactate and glucose levels and serum lactate levels after a 10 min swimming time, as well as ammonia, creatine kinase (CK), blood urea nitrogen (BUN), lactate dehydrogenase (LDH), and glycogen contents after exhaustive swimming time were measured. The exhaustive swimming time of the EMF 200 group was significantly increased (p <0.01). The EMF groups showed significantly low levels of CK, BUN, LDH, and lactate compared with the control group (p <0.05). Increased liver glycogen was observed in the EMF 200 group (p <0.05). These results suggest that EMF can be utilized as an efficacious natural resource for its anti-fatigue effects.

• Journal of Ginseng Research
• /
• v.37 no.2
• /
• pp.210-218
• /
• 2013

Numerous studies have suggested that Korean red ginseng (KRG) extract has various immune modulatory activities both in vivo and in vitro. In this study, we used a mouse model to examine the effects of orally administered KRG extract on immunity against herpes simplex virus (HSV). Balb/c mice were administered with 100, 200, and 400 mg/kg oral doses of KRG extract for 10 d and then vaginally infected with HSV. We found that KRG extract rendered recipients more resistant against HSV vaginal infection and further systemic infection, including decreased clinical severity, increased survival rate, and accelerated viral clearance. Such results appeared to be mediated by increased vaginal IFN-${\gamma}$ secretion. Moreover, increased mRNA expression of IFN-${\gamma}$, granzyme B, and Fas-ligand was identified in the iliac lymph node and vaginal tracts of KRG extract treated groups (200 and 400 mg/kg). These results suggest that the activities of local natural killer cells were promoted by KRG extract consumption and that KRG may be an attractive immune stimulator for helping hosts overcome HSV infection.

• IMMUNE NETWORK
• /
• v.2 no.1
• /
• pp.49-52
• /
• 2002

Background: The possibility that G-CSF recruits leukemic cells from the G0 to S phase, which may lead to a greater susceptibility to cytotoxic drugs, such as ara-C, has been presented in Harada's study. Methods: In this study, we referred to the protocol of Harada et al 1 to try G-CSF combined marrow-ablative chemotherapy and autologous PBSCT, for the treatment of AML patients in CR1 status. Between January 1997 and March 1998, six AML patients (3: children, 3: adults) in CR1 status were autografted and followed up to 3 years. Results: The major regimen related toxicity was composed of mucositis and diarrhea without death. The time of ANC recovery to 500/L and 1,000/L was 11~48 and 16~81 days, respectively. The mean time of platelet recovery to 20,000/L and 50,000/L was 21~233 and 35~370 days, respectively. The platelet recovery time to 50,000/L was markedly prolonged for more than 100 days in four patients (66.7%). Moreover, four patients (66.7%) experienced a relapse of leukemia after transplantation, with a mean interval of 147.5 days after PBSCT. Two patients were in CR status for 53 and 51 months after PBSCT, respectively. Conclusion: The G-CSF combined marrow-ablative chemotherapy and autologous PBSCT resulted in a markedly delayed platelet recovery and no advantages for decreasing the relapse rate of AML. But, further studies will be warranted.

• Journal of Microbiology and Biotechnology
• /
• v.21 no.11
• /
• pp.1151-1158
• /
• 2011

In this study, a galactooligosaccharide (GOS) was synthesized using active ${\beta}$-galactosidase (${\beta}$-gal) inclusion bodies (IBs)-containing Escherichia coli (E. coli) cells. Analysis by MALDI-TOF (matrix-assisted laser desorption/ionization-time of flight) mass spectrometry revealed that a trisaccharide was the major constituent of the synthesized GOS mixture. Additionally, the optimal pH, lactose concentration, amounts of E. coli ${\beta}$-gal IBs, and temperature for GOS synthesis were 7.5, 500 g/l, 3.2 U/ml, and $37^{\circ}C$, respectively. The total GOS yield from 500 g/l of lactose under these optimal conditions was about 32%, which corresponded to 160.4 g/l of GOS. Western blot analyses revealed that ${\beta}$-gal IBs were gradually destroyed during the reaction. In addition, when both the reaction mixture and E. coli ${\beta}$-gal hydrolysate were analyzed by high-performance thin-layer chromatography (HP-TLC), the trisaccharide was determined to be galactosyl lactose, indicating that a galactose moiety was most likely transferred to a lactose molecule during GOS synthesis. This GOS synthesis system might be useful for the synthesis of galactosylated drugs, which have recently received significant attention owing to the ability of the galactose molecules to improve the drugs solubility while decreasing their toxicity. ${\beta}$-Gal IB utilization is potentially a more convenient and economic approach to enzymatic GOS synthesis, since no enzyme purification steps after the transgalactosylation reaction would be required.

• Journal of the Korean Society for Precision Engineering
• /
• v.29 no.7
• /
• pp.711-716
• /
• 2012

The purpose of this study was to investigate how the modulation method affects the effectiveness of eliciting tactile sensations by electrical stimulation. Two methods were employed and the results were compared and analyzed; pulse amplitude modulation (PAM) and pulse width modulation (PWM). Thirty-five healthy subjects participated in the experiments to measure the stimulation intensity that began to elicit a tactile sensation - activation threshold (AT). Constant-current monophasic rectangular pulse trains were employed, and the stimulation intensity was varied from zero until the subject felt any uncomfortable sensation. The step size of the stimulation intensity was 100nC/pulse. After each experiment, the subject described the sensation both quantitatively and qualitatively. The two modulation methods did not make a significant difference as far as the AT values were concerned, but most of the subjects showed 'intra-individual' consistency. Also, it was confirmed that our range of the stimulation parameters enabled us to obtain three major tactile sensations; tickling, pressure and vibration. The results suggested that the stimulation parameters and the modulation type should be selected for each individual and that selective electrical stimulation of the mechanoreceptors needs more diversified researches on the electrode design, multi-channel stimulation protocol, waveforms of the pulse train, etc.

• Asian Pacific Journal of Cancer Prevention
• /
• v.17 no.12
• /
• pp.5251-5256
• /
• 2016

Background: Diagnostic karyotyping analysis is routinely used in acute myeloid leukemia (AML) clinics. Categorization of patients into risk stratified groups (favorable, intermediate and adverse) according to cytogenetic findings can serve as a valuable independent prognostic factor. Method and Material: A retrospective descriptive study was conducted based on the patient records of newly diagnosed non-M3 AML young adult cases undergoing standard 3+7 i.e, Daunorubicin and Ara-C (DA) as remission induction chemotherapy. Diagnostic cytogenetic analysis reports were analyzed to classify the patients into risk stratified groups according to South West Oncology Group criteria and prognostic significance was measured with reference to achievement of haematological remission after 1st induction chemotherapy. Results:A normal karyotype was commonly expressed, found in 47.2% of patients, while 65% (n=39) appeared to have intermediate risk cytogenetics, and 13.3% (n=8) adverse or unclassified findings. Favourable cytogenetics was least frequent in the patient cohort, accounting for only 8.3 % (n=5).The impact of cytogenetic risk groups on achievement of haematological remission was evaluated by applying Pearson Chi-square, and was found to be non-significant (df=12, p=0.256) but when the outcomes of favourable risk groups with intermediate, adverse and unclassified findings compared, results were highly significant (df=6, p=0.000) for each comparison. In patients of the favourable cytogenetic risk group, HR?? was reported in 40% (n=2/5), as compared to 62.2% (n=23/37) in the intermediate cytogenetic risk group, 57.1% (n=4/7) in the adverse cytogenetic risk group and 28.6% (n=2/7) in hte unclassified cytogenetic risk group. Conclusion: Cytogenetic risk stratification for AML cases following criteria provided by international guidelines did not produce conclusive results in our Pakistani patients. However, we cannot preclude an importance as the literature clearly supports the use of pretreatment karyotyping analysis as a significant predictive marker for clinical outcomes. The apparent differences between Pakistani and Western studies indicate an urgent need to develop risk stratification guidelines according to the specific cytogenetic makeup of South Asian populations.

• BMB Reports
• /
• v.54 no.3
• /
• pp.164-169
• /
• 2021

Neuronal growth regulator 1 (NEGR1) is a GPI-anchored membrane protein that is involved in neural cell adhesion and communication. Multiple genome wide association studies have found that NEGR1 is a generic risk factor for multiple human diseases, including obesity, autism, and depression. Recently, we reported that Negr1-/- mice showed a highly increased fat mass and affective behavior. In the present study, we identified Na/K-ATPase, beta1-subunit (ATP1B1) as an NEGR1 binding partner by yeast two-hybrid screening. NEGR1 and ATP1B1 were found to form a relatively stable complex in cells, at least partially co-localizing in membrane lipid rafts. We found that NEGR1 binds with ATP1B1 at its C-terminus, away from the binding site for the alpha subunit, and may contribute to intercellular interactions. Collectively, we report ATP1B1 as a novel NEGR1-interacting protein, which may help deciphering molecular networks underlying NEGR1-associated human diseases.

• CELLMED
• /
• v.11 no.3
• /
• pp.14.1-14.9
• /
• 2021

Background: Cuscuta reflexa Roxb is a member of the Cuscutaceae family, and in Unani medicine, it is known as Aftimoon. It is a parasitic plant that can be found growing abundantly on various host plants in India up to 3000 metres in altitude during the rainy season. Unani physicians have been using it for years to cure a variety of illnesses, including psychiatric illnesses like melancholia, schizophrenia, and epilepsy. It has been used to cure hepatitis, palpitations, and skin disorders, among other things. Objective of the study: To evaluate anti-anxiety effect of Cuscuta reflexa Roxb in Swiss Albino mice of either sex. Materials and Methods: A total of 24 Swiss Albino mice weighing 25-35 g were used in this study. Animals were chosen at random and held in their cages for at least 7 days in a standard setting. Group A was given regular saline as a vehicle, Group B was given a hydro alcoholic extract of the lower dose of the test drug, Group C was given a hydro alcoholic extract of the higher dose of the test drug, and Group D was given the standard drug Diazepam 5 mg/kg orally. Aftimoon as hydro alcoholic extract (200 mg/kg and 400 mg/kg body wt.) was given in single and double doses and observed for 7 days. Results: For each parameter in each category, mean and standard deviations were computed. For multiple group comparisons, a one-way ANOVA was used, followed by Turkey's post hoc test. (p<0.05) was used as the significance standard. Conclusion: These results advocate that the Aftimoon as double dose (400 mg/kg body wt.) revealed anti-anxiety effect similar to standard drug.