Synthetic compounds that are used in the clinic to regulate skin hyperpigmentation, such as arbutin, hydroquinone, and kojic acid, are only moderately effective. But, their use is limited by side effects. As part of an effort to overcome the limitations, we developed resveratrol-enriched rice (RR) using genetic engineering technique. Each of resveratrol and rice has been reported to produce anti-melanogenic effects. Therefore, we hypothesized that RR would show more anti-melanogenic effects than those of resveratrol or rice alone. Anti-melanogenic effect of RR was done by using melan-a mouse melanocytes. The depigmenting efficacy was then observed following topical application of the RR to UVB-stimulated hyperpigmented dorsal skin of guinea pigs. Treatment with RR extract resulted a $21.4{\pm}0.7%$ decrease in tyrosinase expression at melan-a cells. Colorimetric analysis showed a significantly lower depigmenting value by day 9 following treatment with RR in UVB-irradiated guinea pigs the dorsal skin (p<0.01), indicating that RR produced a depigmentation effect. By staining with Fontana-Masson stain, we found that the RR-treated group had more effect histopathologically in epidermal melanin production than resveratrol or rice alone-treated group. RR was associated with reduction in the levels of microphthalmia-associated transcription factor (MITF), and downregulation of tyrosinase and tyrosinase-related protein (TRP-2) expression, leading to inhibit epidermal melanin production by western blot analysis. This study suggests that the resveratrol-enriched rice may be a promising candidate in regulating skin pigmentation with UVB exposure.
A 2-year-old castrated male, Cocker spaniel dog with a history of chronic productive cough for 2 to 3 months and with unsuccessful treatment was referred to Veterinary Medical Teaching Hospital, Seoul National University. On thoracic radiographs, there were alveolar infiltrations at left cranial and right caudal lung fields, and soft-tissue opacity round to oval images at overall lung field. The bronchi were dilated, tortuous and not tapered. Abnormal air was accumulated focally in the caudodorsal lung fields. To scrutinize the soft-tissue opacity image and accumulated air, computed tomography (CT) was done. On CT images, severe cylindrical or tubular bronchiectasis was confirmed. And the soft-tissue opacity images were found in the dilated bilated and thought to complexes of mucous plugs, inflammatory cells, necrotic and fibrotic tissue. The dog was dead next day to the CT scan, so necropsy and histopathologic examination were perfermed. On the histopathology, there were cylindrical bronhiectasis and severe diffuse chronic fibrinous necropurulent bronchitis and bronchopneumonia. In this case, it was difficult to diagnose the bronchiectasis only with radiography due to the concurrent lesions, such as pulmonary infiltrations and mucous plugs, which was identified by computed tomography. Thus, computed tomography is considered as a useful modality to confirm tile bronchiectasis camouflaged by the concurrent lesion.
The purpose of the present study was to produce and characterize a monoclonal antibody against human ${\beta}_2$-adrenergic receptor. Male BALB/c mice were immunized with glutathione S-transferase (GST) fusion protein of the C-terminal portion of the human ${\beta}_2$-adrenergic receptor which was expressed in E.Coli. The immunized splenocytes were fused with myeloma SP2/0-Agl4 cells. The resulting hybridomas were screened for the production of a monoclonal antibody which can recognize human ${\beta}_2$-adrenergic receptor, and then subcloned by limiting dilution. The resulting monoclonal antibody was named as mAb$\beta$CO2. The mono-clonal antibody $\beta$CO2 was determined as IgM subtype and then purified by anti-mouse IgM-agarose affinity chromatography. The results of ELISA, Western blot, and immunocytochemistry showed that mAb$\beta$CO2 recognized human ${\beta}_2$-adrenergic receptor in the ${\beta}_2$-adrenergic receptor-GST fusion protein and human spider-moid carcinoma cell line A431 with highly specific immunoreactivity, The monoclonal antibody $\beta$CO2 may provide useful tools for the study of the $\beta$-adrenergic receptor of human and other species including rats.
Recently, poly-γ-glutamic acid synthetase A (pgsA) has been applied to display exogenous proteins on the surface of Lactobacillus casei or Lactococcus lactis, which results in a surface-displayed component of bacteria. However, the ability of carrying genes encoded by plasmids and the expression efficiency of recombinant bacteria can be somewhat affected by the longer gene length of pgsA (1,143 bp); therefore, a truncated gene, pgsA, was generated based on the characteristics of pgsA by computational analysis. Using murine IL-10 as an exogenous gene, recombinant Lactobacillus plantarum was constructed and the capacity of the surface-displayed protein and functional differences between exogenous proteins expressed by these strains were evaluated. Surface expression of IL-10 on both recombinant bacteria with anchorins and the higher expression levels in L. plantarum-pgsA'-IL-10 were confirmed by western blot assay. Most importantly, up-regulation of IL-1β, IL-6, TNF-α, IFN-γ, and the nuclear transcription factor NF-κB p65 in RAW264.7 cells after stimulation with Poly(I:C) or LPS was exacerbated after co-culture with L. plantarum-pgsA. By contrast, IL-10 expressed by these recombinant strains could reduce these factors, and the expression of these factors was associated with recombinant strains that expressed anchorin (especially in L. plantarum-pgsA'-IL-10) and was significantly lower compared with the anchorin-free strains. These findings indicated that exogenous proteins could be successfully displayed on the surface of L. plantarum by pgsA or pgsA', and the expression of recombinant bacteria with pgsA' was superior compared with bacteria with pgsA.
Xiaohua, Jiang;Zhiwen, Lv;Wenjie, Ding;Ying, Zhang;Feng, Lin
Journal of Electrochemical Science and Technology
/
v.13
no.4
/
pp.431-437
/
2022
As one of the most hazardous and deadliest pathogens, Listeria monocytogenes (LM) posed various serious diseases to the human being, thus designing effective strategy for its detection is of great significance. In this work, by preparing Ti3C2Tx MXenes nanoribbon (Ti3C2TxR) as carrier and selecting thionine (Th) acted simultaneously as signal probe and functional monomer, a LM pathogen-imprinted polymers (PIP) integrated probe electrochemical sensor was design to monitor LM for the first time, that was carried out through the electropolymerization of Th on the Ti3C2TxR/GCE surface in the existence of LM. Upon eluting the templates from the LM imprinted cavities, the fabricated PIP/Ti3C2TxR/GCE sensor can rebound LM cells effectively. By recording the peak current of Th as the response signal, it can be weakened when LM cell was re-bound to the LM imprinted cavity on PIP/Ti3C2TxR/GCE, and the absolute values of peak current change increase with the increasement of LM concentrations. After optimizing three key parameters, a considerable low analytical limit (2 CFU mL-1) and wide linearity (10-108 CFU mL-1) for LM were achieved. In addition, the experiments demonstrated that the PIP/Ti3C2TxR sensor offers satisfactory selectivity, reproducibility and stability.
Rice Oryza sativa accelerated cell death and resistance 1 (OsACDR1) encodes a putative Raf-like mitogen-activated protein kinase kinase kinase (MAPKKK). We had previously reported upregulation of the OsACDR1 transcript by a range of environmental stimuli involved in eliciting defense-related pathways. Here we apply biochemical, gain and loss-of-function approaches to characterize OsACDR1 function in rice. The OsACDR1 protein showed autophosphorylation and possessed kinase activity. Rice plants overexpressing OsACDR1 exhibited spontaneous hypersensitive response (HR)-like lesions on leaves, upregulation of defense-related marker genes and accumulation of phenolic compounds and secondary metabolites (phytoalexins). These transgenic plants also acquired enhanced resistance to a fungal pathogen (Magnaporthe grisea) and showed inhibition of appressorial penetration on the leaf surface. In contrast, loss-of-function and RNA silenced OsACDR1 rice mutant plants showed downregulation of defense-related marker genes expressions and susceptibility to M. grisea. Furthermore, transient expression of an OsACDR1:GFP fusion protein in rice protoplast and onion epidermal cells revealed its localization to the nucleus. These results indicate that OsACDR1 plays an important role in the positive regulation of disease resistance in rice.
Choi, Won Sub;Cho, Jae Hyun;Hwang, Young Il;Jang, Seung Hun;Kim, Dong-Gyu;Jun, Sun-Young;Min, Kwangseon;Lee, In Jae;Lee, Jae Woong;Jung, Ki-Suck
Tuberculosis and Respiratory Diseases
/
v.62
no.5
/
pp.427-431
/
2007
The bronchus-asociated lymphoid tissue(BALT) lymphoma is a low-grade primary malignant lymphoma that originates from bronchus associated lymphoid tissue. A 67-year-old woman was admitted for evaluation of cough, sputum, rhinorrhea which had persisted for one month. Physical examination showed decreased breathing sound on the left upper lung field. High resolution chest computed tomography demonstrated consolidation which showed air-bronchogram and surrounding ground glass opacity in left upper lobe. These findings implicated inactive tuberculosis, organizing pneumonia, or bronchiolo-alveolar carcinoma. The histologic findings from percutaneous needle aspiration biopsy revealed aggregated atypical small lymphoid cells with lymphoepithelial lesions. With immunohistochemical staining, the atypical lymphoid cells reacted positively with CD 20 antibody and negatively with CD 3 antibody. Thus, we could diagnosed her as a patient with BALT lymphoma. After left upper lobectomy, she has been well without recurrence of the disease for 14 months. In this country of Republic of Korea, it was the 1st case of BALT lymphoma surgically treated when histological diagnosis had been done. Based on this case, we wanted to demonstrate the importance of early histological diagnosis and treatment of BALT lymphoma.
Scutellariae Radix (Scu.), one of the immune-regulatory substances, is recognized to play the role in the metabolic process of inflammation, allergy and immunity. It has been traditionally used in the Oriental medicine to treat inflammatory bowel diseases (IBD). The purpose of this study was to evaluate the effects of water extracts of Scutellariae Radix on the spleen lymphocyte immune function in the Balb/c female mice treated with dextran sodium sulfate (DSS) to induce colitis. Water extract of Scutellariae Radix (100 mg/kg) and sulfasalazine (50 mg/kg) were administrated orally for 2 weeks of experimental period. Mice were divided into three experimental groups randomly: DSS group (5% DSS was ad libitum for 5 days) as control group, DSS + Scu. (water extracts of Scutellariae Radix for 2 weeks after 5% DSS was ad libitum for 5 days) as experimental group, and DSS + Sulfasalazine group (Sulfasalazine for 2 weeks after 5% DSS was ad libitum for 5 days) as positive control group. Levels of Ig A, Ig E, CD4$^{+}$, CD8$^{+}$, TNF-$\alpha$ and other cytokines were measured. Treatment of DSS for 5 days induced bowel inflammation and the treatment with Scu. water exteract and sulfasalazine significantly recovered the damage. The length of intestine of DSS group was significantly shorter than that of other groups. The serum and fecal concentration of Ig A of SS + Scu group was higher than those of DSS group. The contents of CD4$^{+}$ T cells was higher in the DSS + Scu. group than the other groups and CD8$^{+}$ T cells was the lowest in DSS + Sulfasalazine group. The Ig A level of cultured supernatant of spleen lymphocyte was the highest, while the Ig E level was the lowest in SS + Scu group. The concentration of TNF-$\alpha$, cytokine secreted from the Th1 cell in the supernatant spleen lymphocyte, was the highest in the DSS group and the lowest in the DSS + Scu. group. The concentration of IFN-${\gamma}$ and ll...-12 was lower in the DSS + Scu. group than those of the other groups. The concentration of IL-4 in the supernatant of spleen lymphocyte was the lowest in the DSS + Scu. group but IL-10 was not significantly different. Based on these findings, water extract of Scutellariae Radix exhibited the inhibitory effect via IL-4 production thereby inhibited the production of Ig E and strengthened immune system, and alleviated injury in DSS- induced colitis mice model.
Kim, Ha-Won;Shim, Mi-Ja;Choi, Eung-Chil;Kim, Byoung-Kak
Archives of Pharmacal Research
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v.20
no.5
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pp.425-431
/
1997
To examine components of Ganoderma lucidum for anti-human immunodeficiency virus (HIV) activity, the aqueous extracts of its basidiocarps were separated into high-molecular-weight (HMF) and low-molecular-weight (LMF) fractions. These fractions were used in XTT [2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide] antiviral assay which can quantitatively measure cytopathic effects of HIV-1 on CEM, human T lymphoblastoid cell line. The CEM cell line added with serial diluted HMF or LMF was cultured in the absence or presence of HIV-1. The results showed that the LMF of the aqueous extract strongly inhibited cytopathic effect of the target cell induced by HIV-1. When two-fold serially diluted LMF ranging from $40.97{\mu}g/ml$4 to 125.00 .mu.g/ml was added to the virus-free culture system, no toxicity on the target cells was detected in all the concentrations tested. However, when it was added to the HIV-infected culture system, the viabilities of the target cell reached a plateau recovering its viabilities to 71.7% and 82.5% in experiment-1 and -2 at 15.60 .mu.g/ml, respectively. The cell viabilities were then gradually decreased but maintained at more than 50% above 31.20 .mu.g/ml concentration. On the contrary, HMF did not prevent any HIV-induced cytopathic effect at any concentrations tested on this cell line. From these results, negligible toxicities were observed by both HMF and LMF of G. luciolum, and recovery of cell viability in HIV infected target cell was induced only by LMF of the carpophores.
Kim, Kyung-Ah;Choi, Seong-Su;Lee, Sang-Bong;Kim, Kyoung-Oak;Park, Kyung-Soon;Shin, Eun-Young;Kim, Wun-Jae;Cha, Eun-Jong
Journal of Sensor Science and Technology
/
v.18
no.6
/
pp.423-431
/
2009
Standard uroflowmetry measures the urine weight using single load cell to evaluate the urinary flow rate. Impact noise should be introduced due to gravity when the urine stream falls down into the container upon the load cell. The present study placed three load cells on the three vertices of a regular triangle and the three signals were ensemble averaged to enhance the signal-to-noise ratio(SNR) regardless of how the urination was made. Simulated urination experiment was performed with three different urine collection methods. In all three methods, SNR of the averaged signal was much higher than each load cell signals. With no urine collection device, the present signal averaging technique resulted in SNR values higher by 10~15 dB than when dual funnels or upper funnel were used to guide the urine stream. Therefore, it was demonstrated that the three point measurement followed by with ensemble averaging could enable accurate uroflowmetric test without any specially made urine collection devices.
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