• The Plant Pathology Journal
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• 제24권2호
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• pp.101-111
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• 2008

The fungal genus Alternaria is comprised of many saprophytic and endophytic species, but is most well known as containing many notoriously destructive plant pathogens. There are over 4,000 Alternaria/host associations recorded in the USDA Fungal Host Index ranking the genus 10th among nearly 2,000 fungal genera based on the total number of host records. While few Alternaria species appear to have a sexual stage to their life cycles, the majority lack sexuality altogether. Many pathogenic species of Alternaria are prolific toxin producers, which facilitates their necrotrophic lifestyle. Necrotrophs must kill host cells prior to colonization, and thus these toxins are secreted to facilitate host cell death often by triggering genetically programmed apoptotic pathways or by directly causing cell damage resulting in necrosis. While many species of Alternaria produce toxins with rather broad host ranges, a closely-related group of agronomically important Alternaria species produce selective toxins with a very narrow range often to the cultivar level. Genes that code for and direct the biosynthesis of these host-specific toxins for the Alternaria alternata sensu lato lineages are often contained on small, mostly conditionally dispensable, chromosomes. Besides the role of toxins in Alternaria pathogenesis, relatively few genes and/or gene products have been identified that contribute to or are required for pathogenicity. Recently, the completion of the A. brassicicola genome sequencing project has facilitated the examination of a substantial subset of genes for their role in pathogenicity. In this review, we will highlight the role of toxins in Alternaria pathogenesis and the use of A. brassicicola as a model representative for basic virulence studies for the genus as a whole. The current status of these research efforts will be discussed.

• The Korean Journal of Physiology and Pharmacology
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• 제2권1호
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• pp.109-117
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• 1998

Role of $Ca^{2+}$/calmodulin complex in intracellular $Ca^{2+}$ mobilization in neutrophils has not been clearly elucidated. In this study, effects of chlorpromazine, trifluoperazine and imipramine on the intracellular $Ca^{2+}$ mobilization, including $Ca^{2+}$ influx, in C5a-activated neutrophils were investigated. Complement C5a- stimulated superoxide production and myeloperoxidase release in neutrophils were inhibited by chlorpromazine, trifluoperazine and imipramine, except no effect of imipramine on myeloperoxidase release. A C5a-elicited elevation of [$Ca^{2+}$]i in neutrophils was inhibited by chlopromazine, trifluoperazine, imipramine, staurosporine, genistein, EGTA, and verapamil but not affected by pertussis toxin. The intracellular $Ca^{2+}$ release in C5a-activated neutrophils was not affected by chlorpromazine and imipramine. Chlorpromazine and imipramine inhibited $Mn^{2+}$ influx by C5a-activated neutrophils. Thapsigargin-evoked $Ca^{2+}$ entry was inhibited by chlorpromazine, trifluoperazine, imipramine, genistein, EGTA and verapamil, while the effect of staurosporine was not detected. The results suggest that $Ca^{2+}$/calmodulin complex is involved in the activation process of neutrophils. The depressive action of calmodulin inhibitors on the elevation of cytosolic $Ca^{2+}$ level in C5a-activated neutrophils appears to be accomplished by inhibition of $Ca^{2+}$ influx from the extracellular medium.

• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.45-53
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• 2010

We have developed an efficient and precise method for genome manipulations in Bacillus subtilis that allows rapid alteration of a gene sequence or multiple gene sequences without altering the chromosome in any other way. In our approach, the Escherichia coli toxin gene mazF, which was used as a counter-selectable marker, was placed under the control of a xylose-inducible expression system and associated with an antibiotic resistance gene to create a "mazF-cassette". A polymerase chain reaction (PCR)-generated fragment, consisting of two homology regions joined to the mazF-cassette, was integrated into the chromosome at the target locus by homologous recombination, using positive selection for antibiotic resistance. Then, the excision of the mazF-cassette from the chromosome by a single-crossover event between two short directly repeated (DR) sequences, included in the design of the PCR products, was achieved by counter-selection of mazF. We used this method efficiently and precisely to deliver a point mutation, to inactivate a specific gene, to delete a large genomic region, and to generate the in-frame deletion with minimal polar effects in the same background.

• ALGAE
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• 제17권2호
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• pp.95-104
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• 2002

Population dynamics of Anabaena and the anatoxin-a concentration were monitored with physicochemical parameters at 3 sites in the lower Naktong River from May to September in 2000. Total 4 species of Anabaena (A. flosaquae, A. smithii, A. ucrainica and A. mucosa) were identified with morphological characterisitcs. Anabaena flos-aquae was most abundant among the populations. The standing crop of Anabaena ranged from 10 to 11,220 cells · $ml^{-1}$ and biomass of Anabaena more 1,000 cells · $ml^{-1}$ was obseved once at St. Mulgeum and St. Seonam, twice at St. Hagueon out of total 9 samplings. There were not significant correlations between the standing crop of Anabaena and other physicochemical parameters such as temperature, nitrate, total nitrogen, phosphate, total phophorus and N/P ratios. The frequency of trichomes with akinetes was low and ranged from 0 to 4% in the total Anabaena population and A. smithii showed highest frequency of 2.8% among all species. The population at St. Seonam showed highest frequency of 1.4% among all sampling sties. The population in September showed the highest frequency of 3.0% among all sampling period. The frequency of trichomes with heterocysts was low and ranged from 1 to 87% inthe total Anabaena population and A. smithii showed highest frequency of 55.1% among all species. The population at St. Mulgeum showed highest frequency of 17.6% among all sampling sites. The population in August showed the highest frequency of 21.4% among all sampling period. The frequency of trichomes with akinetes and/or heterocysts was not related to all the physicochemical parameters of temperature, nitrate, total nitrogen, phosphate, total phosphorus and N/P ratios. The anatoxin-a concentations were determined in algal materials dominated by Microcystis and Anabaena from June though August by derivatization using 7-fluoro-4-nitro-2, 1,3-benzoxadiazole (NBD-F) and HPLC analysis with fluorimetric detection. All the concentrations were below the detection limit of 0.1 ㎍ · $l^{-1}$ in the present study.

• Molecules and Cells
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• 제38권3호
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• pp.243-250
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• 2015

Patterning of the polar axis during the early leaf developmental stage is established by cell-to-cell communication between the shoot apical meristem (SAM) and the leaf primordia. In a previous study, we showed that the DRL1 gene, which encodes a homolog of the Elongator-associated protein KTI12 of yeast, acts as a positive regulator of adaxial leaf patterning and shoot meristem activity. To determine the evolutionally conserved functions of DRL1, we performed a comparison of the deduced amino acid sequence of DRL1 and its yeast homolog, KTI12, and found that while overall homology was low, well-conserved domains were presented. DRL1 contained two conserved plant-specific domains. Expression of the DRL1 gene in a yeast KTI12-deficient yeast mutant suppressed the growth retardation phenotype, but did not rescue the caffeine sensitivity, indicating that the role of Arabidopsis Elongator-associated protein is partially conserved with yeast KTI12, but may have changed between yeast and plants in response to caffeine during the course of evolution. In addition, elevated expression of DRL1 gene triggered zymocin sensitivity, while overexpression of KTI12 maintained zymocin resistance, indicating that the function of Arabidopsis DRL1 may not overlap with yeast KTI12 with regards to toxin sensitivity. In this study, expression analysis showed that class-I KNOX genes were downregulated in the shoot apex, and that YAB and KAN were upregulated in leaves of the Arabidopsis drl1- 101 mutant. Our results provide insight into the communication network between the SAM and leaf primordia required for the establishment of leaf polarity by mediating histone acetylation or through other mechanisms.

• Asian Pacific Journal of Cancer Prevention
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• 제13권10호
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• pp.5063-5067
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• 2012

New molecular markers of cancer had emerged with novel applications in cancer prevention and therapeutics, including for breast cancer of unknown causes, which has a high impact on the health of women worldwide. The purpose of this research was to detemine protein and mRNA expression of synaptic vesicle 2 (SV2) isoforms A, B and C in breast cancer cell lines. Cultured cell lines MDA-MB-231, SKBR3, T47D were lysed and their protein and mRNA expression analyzed by real-time PCR and western blot technique, respectively. SV2A, B proteins were identified in non-tumor (MCF-10A) and tumor cell lines (MDA-MB-231 and T47D) while SV2C only was found in the T47D cell line. Furthermore, the genomic expression was consistent with protein expression for a such cell line, but in MDA-MB-231 there was no SV2B genomic expression, and the SV2C mRNA and protein were not found in the non tumoral cell line. These findings suggest a possible cellular transdifferentiation to neural character in breast cancer, of possible relevance to cancer development, and point to possible use of SV2 as molecular marker and a vehicle for cancer treatment with botulinum toxin.

• 한국동물위생학회지
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• 제19권2호
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• pp.126-138
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• 1996

Lungs from 109 slaughter pigs with gross lesions indicating enzootic Pneumonia of pigs(EPP) and 16 grossly normal lungs, all originating from seven different herds, were subjected to microbiological examinations. The microbiological studies were included both bacterial and mycoplasmal culture. From lungs of 125 slaughter pigs, 87.2% pigs were pneumonia lesions alone or complexly Mycoplasma spp., pasteurella multocidu(p. multocida), Streptococcus spp., and Actinobacillus pleuropneumoniue(A. pleuropneumoniae) were detected in 39.4%, 42.2%, 13.8%, and 3.7% of the pneumonic lungs, respectively. P. multocida was the most frequently isolated organism in pneumonic lungs. Mycoplasmas not isolated organism in 33.9% the pneumonic lungs even If [here are gross lessions mycoplasmas. The amounts of pneumonia in lungs with Mycoplasma spp. alone, a concurrence of Mycoplasma spp. and P. multocida, p. multocida alone, a concurrence of P. multocida and A. pleuropneumoniae, and a concurrence of Mycoplasma spp. and A pleurdpneumoniae were 10.1%, 22.7%, 18.7%, 25%, and 30%, respectively These findings indicated that p. multocida might be involved in the pathogenesis of pneumonia in slaughter pigs. Mycoplasma spp. was also, in this study, associated with higher frequency of pneumonia. The frequency of pigs snout lesion grade 0∼5 inclusive were 27.2%, 28%, 19.2%, 16%, 6.4%, and 3.2% from 125 slaughter pigs. 32(25.6%) pigs were positive and 13~30% in the pigs from seven herds were found to be infected with atrophic rhintis (AR). A total of 46 P. multocida strains In pneumonic lungs were further characterized by capsular serotyping and testing for production of dermonecrotic toxin. 42(91.3%) of strains were capsular A and 4(8.7%) were type D. Out of the type A and type D strains, 86% and 75% were toxigenic, respectively.

• Bulletin of the Korean Chemical Society
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• 제22권12호
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• pp.1361-1365
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• 2001

Recombinant immunotoxins are hybrid cytotoxic proteins designed to selectively kill cancer cells. A divalent immunotoxins, [B3(FabH1)-PE38]2, was constructed by recombining Fab domain of B3 antibody as a cell-targeting domain and Pseudo monas exotoxin A (PE) as a cytotoxic domain. Monoclonal antibody, B3, is the murine antibody (IgG1k) directed against Lewis Y-related carbohydrate antigens, which are abundant on the surface of many carcinomas. Fab fragment of this antibody was used in this study with the modified hinge sequence where last two cysteines out of three were mutated to serine. PE is a 66 kDa bacterial toxin that kills eukaryotic cells by inhibiting protein synthesis with ADP ribosylation of ribosomal elongation factor 2 (EF2). Fc region of B3 antibody was substituted with the truncated form of PE (38 kDa, PE38) on DNA level. [B3(FabH1)-PE38]2 was formed by disulfide bond between cysteines in the modified hinge region of B3(FabH1)-PE38. Each polypeptide for recombinant immunotoxins was overexpressed in Escherichia coli and collected as inclusion bodies. Each inclusion body was solubilized and refolded, and cytotoxic effects were measured. Divalent immunotoxins, [B3(FabH1)-PE38]2, had ID50 values of about 10 ng/mL on A431 cell lines and about 4 ng/mL on CRL1739 cell lines. Control immunotoxins, B3(scFv)-PE40, had ID50 values of about 28 ng/mL on A431 cell lines and about 41 ng/mL on CRL1739 cell lines. Divalent immunotoxins, [B3(FabH1)-PE38]2, had higher cytotoxic effects than B3(scFv)-PE40 control immunotoxins.

• 해양환경안전학회지
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• 제27권1호
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• pp.119-126
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• 2021

본 연구에서는 실내에서 다양한 수온과 염분 조건에서 유독와편모조류 Alexandrium catenella(Group I)의 생장과 함께 마비성패독의 함량을 조사하였다. A. catenella의 수온과 염분 최적생장은 각각 20~30℃과 20~30 psu였으며, 광온성과 광염성종으로 나타났다. A. catenella는 낮은 수온 구간(10℃와 15℃)에서 독함량과 독성이 높게 나타났으며, 염분에 따라서는 차이가 크지 않았다. 따라서 15℃이하의 수온에서 본 종주와 같은 생리적인 특성을 가진 유독와편모조류가 우점한다면, 이매패류는 빠르게 독화될 가능성이 있다. 향후 A. catenella의 출현에 따른 마비성패독 예찰·예보를 위해 다양한 환경변화에 따른 A. catenella의 다른 종주와 상업적인 이매패류 독화에 대한 연구가 더 필요한 것으로 판단된다.

• BMB Reports
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• 제39권3호
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• pp.284-291
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• 2006

In this study, the cDNA of a new peptide from the venom of the scorpion, Buthotus saulcyi, was cloned and sequenced. It codes for a 64 residues peptide (Bsaul1) which shares high sequence similarity with depressant insect toxins of scorpions. The differences between them mainly appear in the loop1 which connects the $\beta$-strand1 to the $\alpha$-helix and seems to be functionally important in long chain scorpion neurotoxins. This loop is three amino acids longer in Bsaul1 compared to other depressant toxins. A comparative amino acid sequence analysis done on Bsaul1 and some of $\alpha$-, $\beta$-, excitatory and depressant toxins of scorpions showed that Bsaul1 contains all the residues which are highly conserved among long chain scorpion neurotoxins. Structural model of Bsaul1 was generated using Ts1 (a $\beta$-toxin that competes with the depressant insect toxins for binding to $Na^+$ channels) as template. According to the molecular model of Bsaul1, the folding of the polypeptide chain is being composed of an anti-parallel three-stranded $\beta$-sheet and a stretch of $\alpha$-helix, tightly bound by a set of four disulfide bridges. A striking similarity in the spatial arrangement of some critical residues was shown by superposition of the backbone conformation of Bsaul1 and Ts1.