Purpose: The purpose of this study is to induce artificial arthritis on rabbit TMJ by injecting collagenase. Materials and Methods: An experimental animal model of arthritis induced by surgical method or intraarticular injection of chemical agent like LDH, papain, ketorolac. Surgical method is complex and needs a long time in inducing arthritis. Intra-articular injection of chemical agent like LDH, papain, ketorolac is simple. But chemical agent like LDH, papain, ketololac needs multiple injections to induce arthritis and mechanism inducing arthritis was known. Collagenase destroys helical domain of type II collagen in extracellular matrix produced by chondrocyte and then induces arthritis. We injected collagenase (0.5, 1.0, 2.0 mg) into the temporomandibular joint of rabbit. In the control group saline was intra-articularly injected. The condylar cartilage, disk and synovia were histologically examined at 1, 2, 4, 6 weeks after the initiation of collagenase injections. Results: Four weeks after injection of 2.0 mg collagenase, we could see histologic change like arthritis. In other groups, we couldn't see arthritis-like change. Conclusion: In our study, we produce arthritis on temporomandibular joint of rabbit by using injection of collagenase in temporomandibular joint of rabbit. And this experimental osteoarthritis is a useful animal model.
Chi, Won-Jae;Youn, Young Sang;Kim, Jong-Hee;Hong, Soon-Kwang
Microbiology and Biotechnology Letters
/
v.43
no.2
/
pp.134-141
/
2015
An agarolytic marine bacterium (H9) was isolated from the coastal seawater of the West Sea, South Korea. The isolate, H9, was gram-negative and rod-shaped with a smooth surface and polar flagellum. Cells grew at 20-30℃, between pH 5.0 and 9.0, and in ASW-YP (Artificial Sea Water-Yeast extract, Peptone) media containing 1-5% (w/v) NaCl. The G+C content was 41.56 mol%. The predominant isoprenoid quinone in strain H9 was ubiquinone-8. The major fatty acids (>10%) were C16:1ω7c (34.3%), C16:0 (23.72%), and C18:1ω7c (13.64%). Based on 16S rRNA gene sequencing, and biochemical and chemotaxonomic characterization, the strain was designated as Pseudoalteromonas sp. H9 (=KCTC23887). In liquid culture supplemented with 0.2% agar, the cell density and agarase activity reached a maximum level of OD = 4.32 (48 h) and OD = 3.87 (24 h), respectively. The optimum pH and temperature for the extracellular crude agarases of H9 were 7.0 and 40℃, respectively. Thin-layer chromatography analysis of the agarase hydrolysis products revealed that the crude agarases hydrolyze agarose into neoagarotetraose and neoagarohexaose. Therefore, the new agar-degrading strain, H9, can be applicable for the production of valuable neoagarooligosaccharides and for the complete degradation of agar in bio-industries.
Rumen microbiology research has undergone several evolutionary steps: the isolation and nutritional characterization of readily cultivated microbes; followed by the cloning and sequence analysis of individual genes relevant to key digestive processes; through to the use of small subunit ribosomal RNA (SSU rRNA) sequences for a cultivation-independent examination of microbial diversity. Our knowledge of rumen microbiology has expanded as a result, but the translation of this information into productive alterations of ruminal function has been rather limited. For instance, the cloning and characterization of cellulase genes in Escherichia coli has yielded some valuable information about this complex enzyme system in ruminal bacteria. SSU rRNA analyses have also confirmed that a considerable amount of the microbial diversity in the rumen is not represented in existing culture collections. However, we still have little idea of whether the key, and potentially rate-limiting, gene products and (or) microbial interactions have been identified. Technologies allowing high throughput nucleotide and protein sequence analysis have led to the emergence of two new fields of investigation, genomics and proteomics. Both disciplines can be further subdivided into functional and comparative lines of investigation. The massive accumulation of microbial DNA and protein sequence data, including complete genome sequences, is revolutionizing the way we examine microbial physiology and diversity. We describe here some examples of our use of genomics- and proteomics-based methods, to analyze the cellulase system of Ruminococcus flavefaciens FD-1 and explore the genome of Ruminococcus albus 8. At Illinois, we are using bacterial artificial chromosome (BAC) vectors to create libraries containing large (>75 kbases), contiguous segments of DNA from R. flavefaciens FD-1. Considering that every bacterium is not a candidate for whole genome sequencing, BAC libraries offer an attractive, alternative method to perform physical and functional analyses of a bacterium's genome. Our first plan is to use these BAC clones to determine whether or not cellulases and accessory genes in R. flavefaciens exist in clusters of orthologous genes (COGs). Proteomics is also being used to complement the BAC library/DNA sequencing approach. Proteins differentially expressed in response to carbon source are being identified by 2-D SDS-PAGE, followed by in-gel-digests and peptide mass mapping by MALDI-TOF Mass Spectrometry, as well as peptide sequencing by Edman degradation. At Ohio State, we have used a combination of functional proteomics, mutational analysis and differential display RT-PCR to obtain evidence suggesting that in addition to a cellulosome-like mechanism, R. albus 8 possesses other mechanisms for adhesion to plant surfaces. Genome walking on either side of these differentially expressed transcripts has also resulted in two interesting observations: i) a relatively large number of genes with no matches in the current databases and; ii) the identification of genes with a high level of sequence identity to those identified, until now, in the archaebacteria. Genomics and proteomics will also accelerate our understanding of microbial interactions, and allow a greater degree of in situ analyses in the future. The challenge is to utilize genomics and proteomics to improve our fundamental understanding of microbial physiology, diversity and ecology, and overcome constraints to ruminal function.
One bacterium, which showed strong antagonistic activity against H. pylori KCCM 41756, was isolated from kimchi. The strain NO1 was designated as Lactobacillus plantarum NO1 based on Gram staining, biochemical properties, and 16S rRNA gene sequencing. The culture medium $(2{\sim}4{\mu}g/ml)$ of Lb. plantarum NO1 reduced $(40{\sim}60%)$ the urease activity of H. pylori KCCM 41756. Lb. plantarum NO1 inhibited the binding of H. pylori to human gastric cancer cell line, AGS cells, by more than 33%. Lb. plantarum NO1 exhibited high viability (maintained initial viable cell count of $10^9CFU/ml$) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastricjuice for 2 h and in 0.3%, 0.5% oxgall for 24 h. Hemolysis phenomena did not observed when Lb. plantarum NO1 was incubated in the blood agar media. We concluded that Lb. plantarum NO1 can be a good candidate as a probiotic, harboring anti-H. pylori activity.
Pogostemonis and Agastachis Herba are the whole of Pogostemon cablin (BLANCO) BENTH or Agastache rugosa (FISCHER et MEYER) O. KUNTZE (family Labiatae) which is produced in all part of Korea and China. This drug is used for removing dampness by means of aromatics in oriental medicine. The standard formula of this drug is important from the viewpoint of the quality control. A characteristic discrimination of internal and external morphological standard in original plants and herbal states of Pogostemonis and Agastachis Herba are as follows. 1. The external characteristics: Pogostemon cablin has hairs and brown-like in stem, elliptical fruit. In the other hand, Agastache rugosa has no hairs and red-like in stem, obovatic trigone fruit. 2. The physical characteristics: Pogostemon cablin is gray in whole, has hairs in stem and numerous hairs of ash in leaf. In the other hand, Agastache rugosa is yellow-green in whole, has no hairs in stem. Specially the latter has deep-green colour and numerous hairs presenting mostly at lower epidermis in leaf. 3. The physical characteristics in currents: Pogostemon cablin is brown, has hairs and round-like stem. In the other hand, Agastache rugosa is green or yellow-green, has no hairs and tetragon in stem. 4. The internal characteristics: Pogostemon cablin has progressed spongy tissue in epidermal cell of leaf and many rank of epidermal cell in stem. In other hand, Agastache rugosa has 1 rank palisade tissue in leaf and few rank of epidermal cell in stem. In the external shape, it was possible that herbs were distinguished according to artificial classification and that same genus-degree of relatedness among herbs could be distinguished by more precise and active observation. In the shape of real herbs, I compared current herbs in market with original herbs which were just collected or were on the course of drying. In addition, it was possible that the internal shape could be identified by using microscope after butanol series. Though it was impossible to make distinction of herbs which are not current in my search contents, this search contents will be a standard for applying herbs in the future. An Additional standard establishment including physiochemical reaction and gene research is required in order to supplement the fault of this search.
In order to provide fundamental data on utilization of dehumidifier in greenhouses, a condensing type dehumidifier using ground water as a coolant was developed and tested dehumidification performance. The developed dehumidifier was applied to greenhouse with fog cooling system and effect of dehumidification on improvement of evaporative cooling efficiency was analyzed. Results of the dehumidifier performance test showed that dehumidification using ground water as a coolant was sufficiently possible in fog cooling greenhouse. When the set point temperature of greenhouse cooling was $32^{\circ}C$ and as temperatures of ground water rose from $15^{\circ}C\;to\;18^{\circ}C,\;21^{\circ}C\;and\;24^{\circ}C$, dehumidification rates decreased by $17.7\%,\;35.4\%\;and\;52.8\%$, respectively. As flow rates of ground water reduced to $75\%\;and\;50\%$, dehumidification rates decreased by $12.1\%\;and\;30.5\%$, respectively. Cooling efficiency of greenhouse equipped with fog system was distinctly improved by artificial dehumidification. When the ventilation rate was 0.7 air exchanges per minute, dehumidification rates of the fog cooling greenhouse caused by natural ventilation were 53.9%-74.4% and they rose up to 75.4%-95.9% by operating the dehumidifier. In case of using the ground water of $18^{\circ}C$ and flow rate of design condition, it was analyzed that whole fog spraying water can be dehumidified even if the ventilation rate is 0.36 exchanges per minute. As a utilization of dehumidifier, it is possible to improve cooling efficiency of fog system in naturally ventilated greenhouses.
The purpose of this study was to investigate the probiotic characteristics and immune activities of selected lactic acid bacterial (LAB) strains as feed additives in livestock. 301 LAB strains isolated from traditional fermented foods were first assessed for their antibacterial activity potential. Of the 301 isolates, five showed antibacterial activity against five livestock pathogens (Esherichia coli KCCM11234, Listeria monocytogens KCTC3710, Salmonella Typhimurium KCTC1926, Staphylococcus aureus KCCM11593, and Shigella flexneri KCTC2517). The probiotic characteristics of the five selected strains were also investigated by antioxidative activity, hemolysis, bile salt hydrolase, acid resistance and bile tolerance. The SRCM102607 strain was found to have superior probiotic properties and was selected for further experimentation. 16S rRNA gene sequencing showed that SRCM102607 is Pediococcus acidilactici, which was labeled as P. acidilactici SRCM102607 (KCCM 12246P). The survival characteristics of P. acidilactici SRCM102607 in artificial gastrointestinal conditions were assessed under exposed acidic (pH 2.0) and bile (0.5% and 1.0%) conditions. P. acidilactici SRCM102607 was also confirmed to have resistance to various antibiotics, including amikacin, gentamicin, vancomycin, and etc. The TNF-α production by P. acidilactici SRCM102607 was 171.86±4.00 ng/ml. These results show that P. acidilactici RCM102607 has excellent potential for use as a probiotic livestock feed additive.
This study examined the effects of SSI argumentation program on the preservice biology teachers' decision-making types and communication ability. The SSI argumentation program was developed based on 'Social Decision-Making & Problem-Solving strategy' and Toulmin's argumentation pattern. The preservice teachers had opportunities of SSI argumentation through small group discussions. They were asked to identify the issues regarding SSI, think of solutions, and make a decision along with claims, warrants, data, and rebuttals. The preservice biology teachers experienced four SSI topics of abortion, euthanasia, gene manipulation, artificial intelligence. The results indicated that the preservice biology teachers significantly improved the communication ability after the intervention, but they did not change their types of decision-making. In addition, after the intervention, the Pearson correlation results indicated that 'the logical type' of decision-making significantly relates to the communication ability(p<.01). The preservice biology teachers mentioned that they improved their ability of considering warrants, data, background information, context, and rebuttals. Further, the preserivce biology teachers mentioned that they became take an interest in socioscientific issues and improved their ability of accepting criticism from others as well as caring about others when they argue each other. This study implicated that the SSI argumentation program has effects on improving personality education in school science.
Milk-related traits (milk yield, fat and protein) have been crucial to selection of Holstein. It is essential to find the current selection trends of Holstein. Despite this, uncovering the current trends of selection have been ignored in previous studies. We suggest a new formula to detect the current selection trends based on single nucleotide polymorphisms (SNP). This suggestion is based on the best linear unbiased prediction (BLUP) and the Fisher's fundamental theorem of natural selection both of which are trait-dependent. Fisher's theorem links the additive genetic variance to the selection coefficient. For Holstein milk production traits, we estimated the additive genetic variance using SNP effect from BLUP and selection coefficients based on genetic variance to search highly selective SNPs. Through these processes, we identified significantly selective SNPs. The number of genes containing highly selective SNPs with p-value <0.01 (nearly top 1% SNPs) in all traits and p-value <0.001 (nearly top 0.1%) in any traits was 14. They are phosphodiesterase 4B (PDE4B), serine/threonine kinase 40 (STK40), collagen, type XI, alpha 1 (COL11A1), ephrin-A1 (EFNA1), netrin 4 (NTN4), neuron specific gene family member 1 (NSG1), estrogen receptor 1 (ESR1), neurexin 3 (NRXN3), spectrin, beta, non-erythrocytic 1 (SPTBN1), ADP-ribosylation factor interacting protein 1 (ARFIP1), mutL homolog 1 (MLH1), transmembrane channel-like 7 (TMC7), carboxypeptidase X, member 2 (CPXM2) and ADAM metallopeptidase domain 12 (ADAM12). These genes may be important for future artificial selection trends. Also, we found that the SNP effect predicted from BLUP was the key factor to determine the expected current selection coefficient of SNP. Under Hardy-Weinberg equilibrium of SNP markers in current generation, the selection coefficient is equivalent to $2^*SNP$ effect.
To investigate the effects of elevated $CO_2$ on the denitrifying bacterial community structure in a wetland soil, dynamics of bacterial community structure was explored in an artificial wetland ecosystem with one of three plant species (T. latifolia, S. lacustris, and 1. effusus) under two levels of $CO_2$(370 ppm or 740 ppm) after 110day incubation. For the analysis of bacterial community structure, functional genes such as nitrite reductase genes (nirS) were PCR-amplified followed by cloning of PCR products and screening by restriction fragment length polymorphism (RFLP). nirS gene fragments were amplified in all analyzed soil samples. Species richness estimated by the number of distinct phylotypes were 83 and 95 in the ambient $CO_2$ treatment and the elevated treatment, respectively. Two phylotypes (type 1 and type 2) were dominant in both of the treatments. Elevated $CO_2$ treatment increased species richness of denitrifying as well as changed a large proportion of denitrifier phylotypes compared to those of the ambient treatment. Overall, the data in this study suggested that the denitrifying communities in the wetland soil are diverse and that the richness of denitrifying bacterial community might be affected by elevated $CO_2$ treatment.
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