• Journal of Life Science
• /
• v.18 no.3
• /
• pp.304-310
• /
• 2008

Heat shock protein 70 (HSP70) is known as molecular chaperone, the fundamental protein participating in various processes, from nascent protein synthesis to protection of proteins during abiotic stresses and developmental programs. However, their biological functions in plants are not yet well known. Here, NtHSP70-1 (AY372069), HSP70 of Nicotiana tabacum induced by heat stress was investigated. To analyze the protective role of NtHSP70-1, transgenic tobacco plants, which constitutively overexpressed NtHSP70-1 as well as contained either the vector alone or having NtHSP70-1 in the antisense orientation, were constructed. The altered NtHSP70-1 levels in plants were confirmed by western blotting and transgenic sense lines exhibited tolerance to heat stress. Seedlings with the constitutively expressed NtHSP70-1 grew as green or healthy plants after heat stress. In contrast, transgenic vector or antisense lines exhibited yellowing of leaves or some delay in growth, which finally led to death. Evaluation of chlorophyll contents of heat-shocked transgenic tobacco seedlings indicated that NtHSP70-1 contributes to thermotolerance by preventing chlorophyll synthesis in plants.

• Plant Biotechnology Reports
• /
• v.4 no.3
• /
• pp.185-192
• /
• 2010

The Arabidopsis ${\omega}$-3 fatty acid desaturase (AtFAD7) catalyzes the synthesis of trienoic fatty acids (TA). A transgenic tobacco line, T15, was produced by a sense AtFAD7 construct and showed a cosuppression-like phenotype, namely extremely low TA levels. The sequence similarity between AtFAD7 and a tobacco ortholog gene, NtFAD7, was moderate (about 69%) in the coding sequences. AtFAD7 siRNAs accumulated at a high level, and both AtFAD7 and NtFAD7 mRNAs are degraded in T15 plants. The low-TA phenotype in T15 was dependent on a tobacco RNA-dependent RNA polymerase6 (NtRDR6). We also produced tobacco RNAi plants targeting AtFAD7 gene sequences. The AtFAD7 siRNA level was trace, which was associated with a slight reduction in leaf TA level. Unexpectedly, this RNAi plant showed an increased NtFAD7 transcript level. To investigate the effect of translational inhibition on stability of the NtFAD7 mRNAs, leaves of the wild-type tobacco plants were treated with a translational inhibitor, cycloheximide. The level of NtFAD7 mRNAs significantly increased after cycloheximde treatment. These results suggest that the translational inhibition by low levels of AtFAD7 siRNAs or by cycloheximide increased stability of NtFAD7 mRNA. The degree of silencing by an RNAi construct targeting the AtFAD7 gene was increased by co-existence of the AtFAD7 transgene, where NtRDR6-dependent amplification of siRNAs occurred. These results indicate that NtRDR6 can emphasize silencing effects in both cosuppression and RNAi.

• Reproductive and Developmental Biology
• /
• v.35 no.4
• /
• pp.475-478
• /
• 2011

In the present study, the effect of cysteine and NT or bisphenol A(BP) on in vitro aturation(IVM) of porcine oocytes were examined. COCs was cultured in NCSU-23 medium supplement with 10% FCS which had previously been covered with mineral oil and equilibrated in a humidified atmosphere of 5% $CO_2$ and 95% air at $38^{\circ}C$. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 0.5~10.0 mM cysteine were $34.0{\pm}3.2%$, $36.0{\pm}3.5%$, $48.0{\pm}3.8%$, $22.0{\pm}3.2%$, respectively. The IVM rate of oocytes cultured in NCSU-23 medium supplement with 0.5~5.0mM NT for 48 hrs were $24.0{\pm}4.2%$, $18.0{\pm}4.9%$, $8.0{\pm}2.2%$, respectively. NT affects oocyte in vitro maturation rate in a dose-dependent. This result were significantly lower than the control group. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 1.0 mM NT+5.0 mM cysteine($38.0{\pm}4.3%$) were significantly higher than that of NT treatment. The IVM rate of oocytes cultured in NCSU-23 medium supplement with 0.05~5.0 mM BP for 48 hrs were $20.0{\pm}4.7%$, $10.0{\pm}5.3%$, $6.0{\pm}3.2%$, respectively. The IVM rate of oocytes cultured in NCSU-23 medium supplement with BP was significantly lower cultured non supplement of BP ($44.0{\pm}3.5%$). BP affects porcine oocyte maturation rate in a dose-dependent manner. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 1.0 mM BP+5.0 mM cycteine ($32.0{\pm}3.2%$) were increased than that of BP treatment.

• The Korean Journal of Physiology and Pharmacology
• /
• v.4 no.3
• /
• pp.227-234
• /
• 2000

Many reports suggest that neurotensin (NT) in the gastrointestinal tract may play a possible role as a neurotransmitter, a circulating hormone, or a modulator of motor activity. NT exerts various actions in the intestine; it produces contractile and relaxant responses in intestinal smooth muscle. This study was designed to investigate the effect of NT on motility of antral circular muscle strips in guinea-pig stomach. To assess the role of $Ca^{2+}$ influx in underlying mechanism, slow waves were simultaneously recorded with spontaneous contractions using conventional intracellular microelectrode technique. At the concentration of $10^{-7}$ M, where NT showed maximum response, NT enhanced the magnitude $(863{\pm}198%,\;mean\;SEM,\;n=13)$ and the frequency $(154{\pm}10.3%,\;n=11)$ of spontaneous contractions. NT evoked a slight hyperpolarization of membrane potential, tall and steep slow waves with abortive spikes $(278{\pm}50%,\;n=4).$ These effects were not affected by atropine $(2\;{\mu}M),$ guanethidine $(2\;{\mu}M)$ and tetrodotoxin (0.2μM). NT-induced contractile responses were abolished in $Ca^{2+}-free$ solution and reduced greatly to near abolition by $10\;{\mu}M$ of verapamil or 0.2 mM of $CdCl_2.$ Verapamil attenuated the effects of NT on frequency and amplitude of the slow waves. Taken together, these results indicate that NT enhances contractility in guinea-pig gastric antral circular muscle and $Ca^{2+}$ influx through the voltage-operated $Ca^{2+}$ channel appears to play an important role in the NT-induced contractile mechanism.

• Molecules and Cells
• /
• v.21 no.1
• /
• pp.141-146
• /
• 2006

We previously showed that NtCDPK1, a tobacco calcium-dependent protein kinase, interacts with and phosphorylates the Rpn3 regulatory subunit of the 26S proteasome, and that both NtCDPK1 and Rpn3 are mainly expressed in rapidly proliferating tissues, including shoot and root meristem. In this study, we examined NtCDPK1 expression in roots using GUS expression in transgenic Arabidopsis plants, and investigated its function in root development by generating transgenic tobacco plants carrying a sense NtCDPK1 transgene. GUS activity was first detected in roots two days after sowing. In later stages, strong GUS expression was detected in the root meristem and elongation zone, as well as the initiation sites and branch points of lateral roots. Transgenic tobacco plants in which NtCDPK1 expression was suppressed were smaller, and their root development was abnormal, with reduced lateral root formation and less elongation. These results suggest that NtCDPK1 plays a role in a signaling pathway regulating root development in tobacco.

• Journal of Veterinary Clinics
• /
• v.38 no.5
• /
• pp.215-220
• /
• 2021

The purpose of this study was to investigate the correlation between vertebral left atrial size (VLAS) and NT-proBNP levels. Sixty-three dogs with myxomatous mitral valve disease (MMVD) were recruited for this study. The MMVD group was classified according to the ACVIM stage based on medical history, clinical signs, and thoracic radiography and echocardiography findings. The N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels were measured using fluorescent immunoassay. VLAS and VHS values were measured using a digital caliper in the right lateral view. There were significant differences in VLAS for each American college of veterinary internal medicine (ACVIM) stage (p < 0.01). As the ACVIM stage increased, it tended to increase. Vertebral heart score (VHS) also showed a tendency to increase with the ACVIM stage (p < 0.01). VLAS was divided into three groups: VLAS <2.6, 2.6≤ VLAS <3.1, and VLAS ≥3.1. NT-proBNP values were significantly different between the three groups (p < 0.01), and VLAS and NT-proBNP values showed a strong positive correlation (r = 0.756, p < 0.01). In conclusion, there were significant differences among ACVIM stages B1 and B2, B1 and C-D, and B2 and C-D groups regarding VLAS. Furthermore, NT-proBNP increased proportionally as VLAS increased. Measurements of VLAS may be helpful in the diagnosis of MMVD in addition to the existing radiological and echocardiographic methods.

• Reproductive and Developmental Biology
• /
• v.29 no.4
• /
• pp.229-234
• /
• 2005

Methods for activation of reconstructed oocytes were examined for the production of nuclear transfer (NT) rat embryos using fetal neural stem cells as donor. Neural stem cells were isolated from Day 14.5 rat fetuses, and the oocytes for recipient cytoplasm were recovered from 4-week old Sprague Dawley rats. After enucleation and nuclear injection, the reconstructed oocytes were immediately exposed to activation medium consisting of 10 mM $SrCl_2$ for 4 h (immediate activation after injection; IAI), or cultured in vitro for $2\~3$ h before activation treatment (injection before activation; IBA). Pre-activated oocytes were also used for NT to test reprogramming potential of artificially activated oocytes. The oocytes were grouped as IIA (immediate injection after activation) and ABI (activation $2\~3$ h before injection). Following NT, the oocytes were cultured in vitro. Development of the NT embryos was monitored at 44 and 119 h after activation. The embryos in groups IAI, mA, and IIA were cleaved to the 2-cell stage at the rates of $36.6\%\;(15/41),\;39.5\%\;(17/43)\;and\;46.3\%$ (25/54), respectively. However, in the ABI group, only one embryo ($1.8\%$, 1/55) was cleaved after activation. After in vitro culture, two NT embryos from IAI group had developed to the morula stage $(4.9\%\cdot2/41)$. However, no morula or blastocyst was obtained in the other groups. These results suggest that immediate activation after injection (IAI) method may be used for the production of rat somatic cell NT embryos.

• Restorative Dentistry and Endodontics
• /
• v.25 no.3
• /
• pp.381-389
• /
• 2000

Lateral condensation with gutta-percha and sealer has been shown to provide an excellent apical seal; however, the lateral condensation technique has demonstrated less favorable apical leakage results in curved canals when compared with straight canals. Placement of endodontic spreaders to within 1 to 2mm of the root canal working length has been advocated for optimum gutta-percha obturation. Due to their stiffness, stainless-steel(SS) spreaders will often fail to achieve this position in curved canals. Newly marketed nickel-titanium(NT) spreaders may offer an advantage in this regard due to the increased flexibility of these instruments. The purpose of this study was to evaluate the effect of NT finger spreader on the sealing ability in lateral condensation technique, compared with conventional SS finger spreader. Twenty four standardized resin models simulating curved canals(30 degree) were randomly placed into 2 groups and instrumented to a #30 master apical file size with Ni-Ti Profile .04 taper series using step down technique. Each groups was obturated with standardized gutta-percha cone by standard lateral condensation technique using SS finger spreader, NT finger spreader. And then, each model was sectioned horizontally with microtome at 1, 2, 3, 4, 5mm levels from the apex. At each of 5 levels, ratio of the area of gutta-percha was obtained by calculating the area of gutta-percha to the total area of the canal. The data collected were then analyzed statistically using a t test for independent samples. The results as follows ; 1. The total mean ratio of area of gutta-percha was 89.20${\pm}$7.00(%) for SS spreader group. 92.20${\pm}$5.17(%) for NT spreader group. There was statistically significant difference between each group(p<0.05). 2. At 3mm level, the mean ratio of area of gutta-percha was 88.32${\pm}$5.41(%) for SS spreader group, 95.25${\pm}$2.60(%) for NT spreader group. There was statistically significant difference between each group(p<0.05). At 1,2,4mm levels, NT spreader group showed greater mean ratio of area of gutta-percha than SS spreader group, too. But there was no statistically significant difference. 3. At 5mm level, the mean ratio of area of gutta-percha was 91.83${\pm}$3.42(%) for SS spreader group, 87.91${\pm}$3.68(%) for NT spreader group. There was statistically significant difference between each group(p<0.05). This study concluded that the NT spreader demonstrated somewhat favorable apical sealing effect than SS spreader in prepared curved canals. The clinical use of NT spreaders may enhance our ability to create better apical seals in curved canals, but further studies in this area will help clarify some of the remaining areas with which practitioners are concerned, such as compaction forces exerted by NT spreaders.

• Journal of Veterinary Clinics
• /
• v.28 no.3
• /
• pp.273-279
• /
• 2011

Botulinum toxin type A (BoNT/A) has been proven to be a safe and effective treatment for wrinkles in human. As well as the anti-wrinkle effects, the anti-pruritic effect of BoNT/A has been revealed from several researches for new therapy. The aim of this study was to investigate the anti-pruritic effect of BoNT/A against histamine-induced pruritus on canine skin. Five clinically healthy beagles were included in the study. All dogs were received 0.05 ml (5 Unit) of BoNT/A on the right dorsal thoracic region as an experiment and the same volume of saline solution was injected on the left dorsal thoracic region as a control, respectively. Intradermal histamine injections were performed four times (before treatment and days 1, 3 and 7 after BoNT/A injection). The severity of pruritus, the diameter and thickness of wheal, the erythema index and cutaneous temperature were assessed. The severity of pruritus was reduced on BoNT/A treated sides, compared with saline treated sides (p < 0.05). BoNT/A decreased wheal size, in both diameter and thickness (p < 0.05). Although, erythema index of both sides were increased after first histamine injection, BoNT/A treated sides showed the low-value as compared with saline treated sides. Cutaneous temperature was decreased significantly on BoNT/A treated sides. These results indicates that BoNT/A reduce histamine-induced pruritus on canine skin and suggested a possibility of application of BoNT/A for local intractable dermatologic problem in dogs.

• Korean Journal of Biological Psychiatry
• /
• v.4 no.2
• /
• pp.234-236
• /
• 1997

Though initial report from Japan showed positive association of schizophrenia with dinucleotide repeat polymorphism in the NT-3 gene, subsequent studies showed mixed results. Therefore we conducted a replication study with Korean schizophrenics and matched controls who share similar ethnic background with Japanese population. The frequency of allele of dinucleotide repeat at 147 base pairs in the NT-3 gene was slightly increased, however, failed to reach statistical significance(${\chi}^2$=1.884, df=1, p<0.170) between the two groups. These findings do not support an association of NT-3 gene polymorphism with schizophrenia in Korean sample.