Upregulation of relaxin receptors in the PDL by biophysical force.

OBJECTIVES: We have previously reported that relaxin (Rln) expression from the ovary is upregulated by orthodontic tooth movement. This study was performed to test the hypothesis that Rln family peptides (Rxfps), the G-protein-coupled Rln receptor, is induced in periodontal ligament (PDL) cells to modulate the molecules involved in periodontal tissue remodeling while applying biophysical force. MATERIALS AND METHODS: Rats were implanted with orthodontic appliances to investigate changes to Rxfps in vivo. An in vitro biophysical force analysis was performed to measure the level of Rxfp 1 messenger RNA (mRNA) in primary human PDL cells. RESULTS: The levels of Rxfp 2 transcription and translation increased in a time-dependent manner during tooth movement. Rxfp 2 was localized in the PDL by immunofluorescence. In vitro analyses revealed that the level of Rxfp 1 mRNA in PDL cells increased significantly with both compression and tension force. The levels of matrix metalloproteinase (MMP)-1, MMP-2, interleukin-6, and vascular endothelial growth factor mRNA, which are important for periodontal tissue remodeling, also changed under force application and Rln treatment. CONCLUSIONS: PDL cells responded to Rln to modulate effector molecules for periodontal tissue remodeling by upregulating Rxfps expression under a biophysical force. CLINICAL RELEVANCE: Rln and Rxfps may serve as a PDL turnover molecule complex to control orthodontic tooth movement.

Responses of blood glucose, insulin, glucagon, and Fatty acids to intraruminal infusion of propionate in hanwoo.

This study was carried out to investigate the effects of intraruminal infusion of propionate on ruminal fermentation characteristics and blood hormones and metabolites in Hanwoo (Korean cattle) steers. Four Hanwoo steers (average body wt. 270 kg, 13 month of age) equipped with rumen cannula were infused into rumens with 0.0 M (Water, C), 0.5 M (37 g/L, T1), 1.0 M (74 g/L, T2) and 1.5 M (111 g/L, T3) of propionate for 1 hour per day and allotted by 4×4 Latin square design. On the 5th day of infusion, samples of rumen and blood were collected at 0, 60, 120, 180, and 300 min after intraruminal infusion of propionate. The concentrations of serum glucose and plasma glucagon were not affected (p>0.05) by intraruminal infusion of propionate. The serum insulin concentration at 60 min after infusion was significantly (p<0.05) higher in T3 than in C, while the concentration of non-esterified fatty acid (NEFA) at 60 and 180 min after infusion was significantly (p<0.05) lower in the propionate treatments than in C. Hence, intraruminal infusion of propionate stimulates the secretion of insulin, and decreases serum NEFA concentration rather than the change of serum glucose concentration.

Effects of Coconut Materials on In vitro Ruminal Methanogenesis and Fermentation Characteristics.

The objective of this study was to evaluate the in vitro effects of coconut materials on ruminal methanogenesis and fermentation characteristics, in particular their effectiveness for mitigating ruminal methanogenesis. Fistulated Holstein cows were used as the donor of rumen fluid. Coconut materials were added to an in vitro fermentation incubated with rumen fluid-buffer mixture and timothy substrate for 24 h incubation. Total gas production, gas profiles, total volatile fatty acids (tVFAs) and the ruminal methanogens diversity were measured. Although gas profiles in added coconut oil and coconut powder were not significantly different, in vitro ruminal methane production was decreased with the level of reduction between 15% and 19% as compared to control, respectively. Coconut oil and coconut powder also inhibited gas production. The tVFAs concentration was increased by coconut materials, but was not affected significantly as compared to control. Acetate concentration was significantly lower (p<0.05), while propionate was significantly higher (p<0.05) by addition of the coconut materials than that of the control. The acetate:propionate ratio was significantly lowered with addition of coconut oil and coconut powder (p<0.05). The methanogens and ciliate-associated methanogens in all added coconut materials were shown to decrease as compared with control. This study showed that ciliate-associated methanogens diversity was reduced by more than 50% in both coconut oil and coconut powder treatments. In conclusion, these results indicate that coconut powder is a potential agent for decreasing in vitro ruminal methane production and as effective as coconut oil.

The Effect of Plant Extracts on In-vitro Ruminal Fermentation, Methanogenesis and Methane-related Microbes in the Rumen.

The effect on methanogens attached to the surface of rumen ciliate protozoa by the addition of plant extracts (pine needles and ginkgo leaves) was studied with particular reference to their effectiveness for decreasing methane emission. The plant extracts (pine needles and ginkgo leaves) were added to an in vitro fermentation incubated with rumen fluid. The microbial population including bacteria, ciliated-associated methanogen, four different groups of methanogens and Fibrobacter succinogenes were quantified by using the real-time PCR. Gas profiles including methane, carbon dioxide and hydrogen, and runinal fermentation characteristics were observed in vitro. The methane emission from samples with an addition of individual juices from pine needles, ginkgo leaves and 70% ethanol extract from ginko leaves was significantly lower (p<0.05, 27.1, 28.1 and 28.1 vs 34.0 ml/g DM) than that of the control, respectively. Total VFAs in samples with an addition of any of the plant extracts were significantly lower than that of the control (p<0.05) as well. The order Methanococcales and the order Methanosarcinales were not detected by using PCR in any incubated mixtures. The ciliate-associated methanogens population decreased from 25% to 49% in the plant extacts as compared to control. We speculate that the supplementation of juice from pine needles and ginkgo leaves extract (70% ethanol extract) decreased the protozoa population resulting in a reduction of methane emission in the rumen and thus inhibiting methanogenesis. The order Methanobacteriales community was affected by addition of all plant extracts and decreased to less than the control, while the order Methanomicrobiales population showed an increase to more than that of the control. The F. succinogenes, the major fibrolytic microorganism, population in all added plant extracts was increased to greater than that of the control. In conclusion, pine needles and ginkgo leaves extracts appear to have properties that decrease methanogenesis by inhibiting protozoa species and may have a potential for use as additives for ruminants.

Effects of Supplementing Microbially-fermented Spent Mushroom Substrates on Growth Performance and Carcass Characteristics of Hanwoo Steers (a Field Study).

This study evaluated the effect of dietary supplementation of microbially-fermented spent mushroom substrates (MFSMS) on weight gain, carcass characteristics, and economic efficiency of Hanwoo steers. Highly cellulolytic bacteria (Enterobacter spp. and Bacillus spp.) isolated from spent mushroom substrates (SMS) stacks were inoculated (1% v/v) into the SMS, which was anaerobically fermented and fed to the steers for 12.6 months during the growing and fattening periods. Growing Hanwoo steers were assigned to the control group without supplementation of Microbially-fermented SMS (MFSMS), to a treatment group with 50% of MFSMS (1/2 of the ad libitum group), and to a treatment group with ad libitum access to SMS (the ad libitum group). All the groups were fed the formulated feed and rice straw. The voluntary intake (DM basis) of MFSMS was 1.6 kg/d during the growing period and 1.4 kg/d during the fattening period. The voluntary rice straw intake decreased by 6 to 11%, but the total voluntary DMI increased by 7 to 15% with MFSMS fed. The increased DMI with MFSMS supplementation resulted in a tendency of increased (p = 0.055) live weight gain by 8 to 12% compared with the control group. At slaughtering, the supplementation of MFSMS increased (p<0.05) the ribeye area by an average of 10 cm(2). In conclusion, feeding MFSMS improved growth performance and carcass traits of Hanwoo steers and could successfully replace a part of conventional roughage such as rice straw commonly used in Asian countries.

Inhibitory effect of mast cell-mediated immediate-type allergic reactions in rats by spirulina.

We investigated the effect of spirulina on mast cell-mediated immediate-type allergic reactions. Spirulina dose-dependently inhibited the systemic allergic reaction induced by compound 48/80 in rats. Spirulina inhibited compound 48/80-induced allergic reaction 100% with doses of 100-1000 microg/g body weight, i.p. Spirulina (10-1000 microg/g body weight, i.p.) also significantly inhibited local allergic reaction activated by anti-dinitrophenyl (DNP) IgE. When rats were pretreated with spirulina at a concentration ranging from 0.01 to 1000 microg/g body weight, i.p., the serum histamine levels were reduced in a dose-dependent manner. Spirulina (0.001 to 10 microg/mL) dose-dependently inhibited histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. The level of cyclic AMP in RPMC, when spirulina (10 microg/mL) was added, transiently and significantly increased about 70-fold at 10 sec compared with that of control cells. Moreover, spirulina (10 microg/mL) had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha production. These results indicate that spirulina inhibits mast cell-mediated immediate-type allergic reactions in vivo and in vitro.

Expressed sequence tags of radish flower buds and characterization of a CONSTANS LIKE 1 gene.

Expressed sequence tag (EST) analysis was conducted for young flower buds of radish plants. Among a total of 66 ESTs examined, 40 showed a significant similarity to previously identified genes. Twenty-eight ESTs were similar to proteins identified in other plants, 11 were similar to eukaryotic proteins other than plants, and one was similar to a prokaryotic protein. Four clones were selected for further studies. EST clone 81, which showed a homology to germin-like proteins was expressed more abundantly in leaves and roots as compared to flower buds. Clone 105 was highly homologous to the translation inhibitor protein and was expressed in all three organs, but the expression level was higher in flower buds and roots. Another EST clone, 133, which shared a significant similarity with the Ran-binding protein, hybridized to two different size transcripts that were detectable only in flower buds. Clone 39 was a homolog of CONSTANS, which is a gene involved in controlling the flowering time in Arabidopsis. The cDNA clone of EST clone 39 containing the entire open reading frame was obtained and designated as RsCOL1 (Raphanus sativus CONSTANS LIKE 1). It was 1049 bp long and contained an open reading frame of 307 amino acid residues (calculated molecular mass = 33.1 kDa). The RsCOL1 protein contained two putative zinc finger motifs in the amino terminal region which were 59% identical to the corresponding region of the Arabidopsis CO protein. The radish protein also contained a predicted nuclear localization domain in the carboxyl terminal region which was 87% identical to the corresponding region of CO. DNA blot analysis revealed that the radish genome contained several genes similar to RsCOL1. RNA blot analysis showed that RsCOL1 was strongly expressed in flower buds at the early bolting stage, and the expression level declined as the flower bud matured. The transcript was also detectable in leaves and roots. In mature flowers, the RsCOL1 transcript was present primarily in carpels.

Expression of a cDNA encoding Phytolacca insularis antiviral protein confers virus resistance on transgenic potato plants.

To develop an antiviral agent and virus-resistant plants, a cDNA clone encoding Phytolacca insularis antiviral protein (PIP) was isolated from a cDNA library constructed with poly(A)+ RNA purified from leaves of P. insularis. The PIP cDNA contains an open reading frame encoding 307 amino acids. The deduced amino acid sequence includes a putative signal sequence of 22 amino acids at the N-terminus. The amino acid sequence of PIP shares 84% homology with that of the pokeweed antiviral protein (PAP). In addition, the mature PIP exhibits the conserved putative active site found in other ribosome-inactivating proteins (RIPs). Recombinant PIP (rPIP) synthesized in Escherichia coli inhibits protein synthesis in vitro in rabbit reticulocyte lysate through the N-glycosidase activity in a similar manner with other RIPs. Local lesion assays with purified rPIP revealed that it inhibits infection of various viruses to plants. Transgenic potato plants expressing the PIP cDNA under the control of the cauliflower mosaic virus 35S promoter are resistant to viruses, such as potato virus X, potato virus Y, and potato leafroll virus. These results suggest that the PIP cDNA could be used for the development of an antiviral agent and transgenic plants resistant against a broad spectrum of plant viruses infecting through both mechanical and aphid transmission.

Characterization of MADS box genes from hot pepper.

The cDNA clone, CanMADS1, was isolated from young flower buds of the hot pepper (Capsicum annuum L.) by screening a cDNA library using the OsMADS1 rice MADS-box gene as a probe. We used a yeast two-hybrid screening method to investigate interaction partners of the protein product of CanMADS1. A MADS-box gene, CanMADS6, was isolated from young flower buds using the region containing the K domain and 15 amino acid residues of the C-terminal region of CanMADS1 as a bait. CanMADS1 and CanMADS6 showed high amino acid sequence similarities to members of the AGL2 subfamily and the SQUA subfamily, respectively. CanMADS1 transcript was expressed in flower buds and fruits, and the transcription signal was the strongest in the stage of the fruit set (2 d after anthesis). CanMADS6 showed the same expression pattern as CanMADS1. CanMADS1 and CanMADS6 were not expressed in leaves. These results suggest that a regulatory role for flower and fruit development of the hot pepper may be accomplished through an interaction of the protein products of the two MADS-box genes, CanMADS1 and CanMADS6.

Structure and expression of the AWI 31 gene specifically induced by wounding in Arabidopsis thaliana.

Differential screening of an Arabidopsis cDNA library constructed from the plant tissues harvested 1 h after wounding resulted in the isolation of wound-inducible cDNA clones (Kim et al., 1994). The cDNA clones could be broadly classified into two groups according to the expression time of their transcripts. Nine clones from the 10 different wound-inducible cDNAs were rapidly induced, reaching a maximum level in approximately 1-1.5 h and then were progressively reduced after wounding. The cDNA clone AWI 31 showed steady accumulation of the transcripts and reached the maximum value at a later time point of 2.5 h and then started to decline. The corresponding gene of the AWI 31 in which the coding region was interrupted by an intron, had an open reading frame that predicted a protein of 386 amino acids. However, the gene product did not show any significant homology to other known proteins in the database. Northern hybridization study using the cDNA probe revealed that the gene was not regulated by other environmental stresses such as drought, high salt, low temperature, or a DPE herbicide treatment, indicating that the cDNA clone AWI 31 was specifically induced by wounding.

Cloning of a cDNA encoding phospholipase D from Pimpinella brachycarpa.

Phospholipase D (PLD, EC 3.1.4.4) has been known to be related to various cellular processes in plants. To gain an understanding of the property of the enzyme in Pimpinella brachycarpa, the cDNA of the enzyme was isolated by PCR with degenerate primers, cDNA library screening, and 5' RACE. The full-length PLD cDNA is 2859 bp long and contains an open reading frame of 2424 bp coding for a polypeptide of 808 amino acids. The deduced enzyme has a calculated molecular mass of 91.7 kDa and pI of 5.86. The percent identity and similarity values of P. brachycarpa PLD with those of other PLDs in plants are 70 approximately 78 and 84 approximately 95, respectively. It was identified that PLD from P. brachycarpa has HQKIVVVD and HAKMMIVD sequences which were homologous with a duplicated HXKXXXXD motif that has been conserved in PLDs from plants, animals, and yeast. Based on the analysis of amino acid similarity, it is believed that PLD from P. brachycarpa is an alpha form which is distinct from PLD beta reported recently. The N-terminus is homologous to the C2 domain which is present in a number of different proteins involved in signal transduction and membrane trafficking in animals. Southern and northern blot analyses indicated that PLD was expressed from one copy of PLD gene in the genome of P. brachycarpa.

Co-administration of toluene and xylene antagonized the testicular toxicity but not the hematopoietic toxicity caused by ethylene glycol monoethyl ether in Sprague-Dawley rats.

Occupational painters are exposed to ethylene glycol monoethyl ether (EGEE), a widely used emulsifying solvent known to cause testicular degeneration and bone marrow depression, together with toluene (TOL) and xylene (XYL) as a mixture. In the previous study (Chung et al., Tox. Lett. 104:143, 1999), testicular atrophy caused by EGEE (200 mg/kg) was shown to be antagonized by co-administration of TOL (250 mg/kg) and XYL (500 mg/kg). This study was conducted to provide histological support for the previously observed antagonistic protective effect of TOL + XYL on EGEE inducible testicular toxicity and to determine whether a similar antagonistic effect can be demonstrated against the EGEE derived hematopoietic toxicity. Compared to the extent of seminiferous tubule degeneration caused by EGEE (150 mg/kg, six times per week for 4 weeks), testes of rats given co-administration of TOL (250 mg/kg) + XYL (500 mg/kg) showed dramatically reduced tubular degeneration. Hyperplasia of Leydig cells in the interstitium was observed in both EGEE and EGEE + TOL + XYL-treated rats. Although a minimal dose of EGEE causing testicular atrophy was used, WBC and platelet counts were decreased significantly. In the TOL + XYL-treated control group, the WBC and platelet counts were not decreased. However, the bone marrow depression caused by EGEE was not reversed by the combined administration of TOL + XYL. In all experimental groups (EGEE alone, TOL + XYL, EGEE + TOL + XYL), plasma levels of creatinine and alkaline phosphatase were significantly decreased. In addition to the marked testicular atrophy, EGEE also decreased the weights of adrenal glands and epididymis. In conclusion, while the testicular degeneration caused by EGEE was antagonized by TOL + XYL, the EGEE derived hematopoietic suppression was not reversed.

Reproductive toxicity of 2-bromopropane in Sprague Dawley rats.

OBJECTIVES: In an effort to clarify the mass intoxication of workers at an electronic company in Korea, the possible causative chemical for reproductive toxicity, 2-bromopropane (2BP), was investigated. METHODS: 2BP was tested through the use of repeated dose experiments among male Sprague Dawley rats. Ten rats were assigned to each treatment group. Vehicle control olive oil and 2BP concentrations of 125 mg/kg, 250 mg/kg, and 500 mg/kg were injected into the intraperitoneum on 28 consecutive days. RESULTS: The rats showed significant decreases in body weight depending on the 2BP dose. The right and left testes showed typical weight loss depending on the dose of 2BP. The red blood cells, hemoglobin, and hematocrit showed some degree of decline with the high dose. The amount of hemoglobin, the mean platelet volume, the number of white blood cells, and the number of lymphocytes decreased significantly with the high dose, while the number of granulocytes and monocytes had a tendency to decrease depending on the dose of 2BP. The histopathology of the testes treated with the middle and high 2BP dose showed a typical patch appearance with severely depleted atrophic tubules, exhibiting germ cell necrosis of spermatogonia and spermatocytes in the seminiferous tubules. Leydig cell hyperplasia or hypertrophy in the interstitial tissue was also noticeable. The epididymis showed some degree of atrophy with vacuolization of the epididymal epithelium. CONCLUSIONS: The testes are the main target organs tested for 2BP toxicity. 2BP also affect the hematopoietic system and thus induces leukopenia and normocytic anemia. Besides the reproductive organs and the hematopoietic system, no significant toxicity has been found.

A fast fractal decoding algorithm based on the selection of an initial image.

In this correspondence, we show that the number of iterations required for the convergence of the fractal image decoding algorithm can be reduced by selecting a suitable initial image. We consider fractal decoding to have two components, namely, "DC" and "AC". Our initial image is an approximation of the attractor of DC decoding, which is a good estimation of the "range-averaged" image, i.e., the image obtained by replacing all pixel intensities in a given block by the mean value of the block. From the simulation, it is demonstrated that the decoding speed is greatly improved.

Inhibition of stem cell factor- and nerve growth factor-induced morphological change by wortmannin in mast cells.

Recombinant murine stem cell factor (rmSCF) or recombinant murine nerve growth factor (rmNGF) induced the morphological change of large numbers of rat peritoneal mast cells (RPMC). We investigated the role of phosphatidylinositol 3'-kinase (PI3-kinase) in receptors-mediated morphological change in RPMC. Exposure of RPMC to PI3-kinase inhibitor, wortmannin, before the addition of rmSCF and rmNGF antagonized those factors-induced morphological change. These results suggest that the Pl3-kinase is involved in the signal transduction pathway responsible for morphological change following stimulation of rmSCF and rmNGF and that wortmannin blocks these responses.

Effects of 2-bromopropane on the female reproductive function in Sprague-Dawley rats.

2-Bromopropane (2BP) has been implicated to be the reason for the mass intoxication of workers at an electronic company in Korea. A case series study indicated that 2BP was the possible causative chemical for reproductive toxicity, causing severe anemia accompanied by amenorrhea among female workers, and azoospermia or oligospermia among male workers. To clarify the effect of 2BP on the female reproductive function, repeated doses of 2BP were tested on female Sprague-Dawley rats for 21 days. Ten rats were assigned to each treatment group. The rats were maintained in a 12 hr: 12 hr light-dark cycle and vaginal smears were monitored daily for 3 cycles. After the rats had completed 3 estrous cycles, vehicle control olive oil, 300 mg, 600 mg, and 900 mg/kg of 2BP were injected into intraperitoneum for 14 days. The female rats were then mated with male rats on a 1:1 ratio basis. The treatment continued for an additional 7 days. Rats treated with 2BP experienced a significant decrease in body weight gain depending on the dose of 2BP. The estrous cycles of the rats continued at a normal duration of time before the initiation of treatment, showing 4.32 days for the control group, 4.79 days for the low dose, 4.63 days for the middle dose, and 4.75 days of estrous cycle for the high dose group. 2BP treatment, however, induced a significant delay of the estrous cycle in the high dose treated group, showing 11.1 +/- 3.82 days of the estrous cycle. 2BP decreased the fertility and tended to decrease in the number of pups born, depending on the dose. A 900 mg/kg treatment of 2BP decreased ovarian weight, but 2BP did not affect the length of gestation. Our results indicated that 2BP seemed to be the causative agent for amenorrhea observed in female workers.

A false presence of bifid mandibular canals in panoramic radiographs.

OBJECTIVES: This study was performed in order to verify bifid mandibular canals revealed from panoramic radiographic results. METHODS: 1000 panoramic radiographs from dental patients and the panorama, cone beam CT (CBCT) and micro-CT from 40 dry mandibles were examined for bifid mandibular canals. The results were confirmed by a stereoscopic and histological examination of the cross-sectioned mandibles. RESULTS: The prevalence of bifid canals detected from the panoramic radiographs was 0.038. The panoramic radiographs from one dry mandible showed two separate radiolucent mandibular canal-like structures delineated by radio-opaque lines. However, a stereoscopic and histological examination of a cross-section of the mandible showed that only one canal was a true canal containing neurovascular bundles: the other was false, reflecting merely a bony trabecular pattern. CONCLUSIONS: The presence of bifid mandibular canals determined by panoramic radiography should be judged with great caution in relation to dental surgery.