The authors made a clinical study of 80 cases of empyema who were diagnosed and treated at department of chest surgery, St. Mary`s Hospital, Chatholic Medical College, during the period of May.l964 through April.1969 and compared the empyema of infant and children with that of adults. 1. In age and sex ditribution, infant was 6 cases, childhood 22 cases and adult 52 cases. The ratio of male to female was 2.2:1. There`s a little difference in infant-childhood but prominence of males over females in adults was being 3. 3:1, in its ratio. 2. The cardinal symptoms were cough [61.3%], fever [60.0%] and dyspnea [52.8%]. The leukocytosis were observed in 83.7% of all cases, 96.2% of infant-childhood and 76.9% of adults. The hemoglobin level showed subnormal in 82.1% of infant-childhood and in 55.8% of adults. 3. Most frequent lesion to predisposing factor of empyema was pneumonia [43.7%],being prominent in infants children [64.3%] to that of adult 4. The Pathogenic organism by culture in 75 cases of empyema were staphylococuss [48%], streptococuss[9.3%], Gram[-] bacilli [9.3%], Klebsiella[2.7%], pneumococcus[4.0%], E. coli [5.4%] and no growth 21.3% in over all. Among the cases of empyema. staphlocal origin was 62.9% in infant-childfood and 39.6% adults. 5. Staphylococci were most susceptible to erythromycin [86. 1%], Kanamycin [75.0%], albamycin [61.7%] and neomycin [52.8%] but most resistant to penicillin, Chtoramphenicol and terramycin. 6. In the treatment of empyema, of 53 cases were closed thoracotomy drainage and the remainder of cases by open thoracotomy, decortication, thoracoplasty and pleuropneumonectomy. we could attain favourable results by only the closed thoracotomy in infant-childhood, 28 cases. 7. The mortality rate was 6.3% in over all; adult 3 cases, infant and children 2 cases. 3 cases of these, were due to staphylococcal infection.
The cell growth inhibitor effect of cervical cancer cells was investigated by liposome mediated transfection (pRcCMVp53/lipofectin) and by transfection using adenovirus (AdCMVp57). The papilloma virus cancer cell lines we used in this study were HPV16 positive, having inhibiter gene, wild p53 gene, CaSki, SiHa, HPV18 positive HeLa, HeLaS3 and HPV negative C33A, HT3. LacZ gene of E.coli was used as the marker gene for the transfection efficiency. The effect on the inhibition of tumor cell growth was measured by cell count and cell viability though ELISA analysis and MTT assay. The inhibition of tumor cell growth was confirmed by measuring each assay for six days, comparing with the normal control cell growth. The cell growth of cervical cancer calls by transfection was significantly reduced and showed tittle differences among the cell lines. To eliminate the potential problem of Ad(adenovirus) contamination during rAAV production, rAAV can be produced by a triple transfection of vector plasmic, packaging plasmid, and adenovirus helper plasmid. To examine the helper functions of Ad plasmids on the production of rAAV vector, we carried out cotransfection of three plasmids, AAV vector, packaging construct, and Ad helper plasmids. The optimized transfection condition for calcium phosphate method is 25ug of total DNA per 10-cm-diameter plate of 293 cell. We found that rAAV yields peaked at 48hr after Ad infection. The titer of rAAV was measured by the dot blot analysis to measure the number of particles/ml based on the quantification of viral DNA. Recent1y, Kombucha(fungi) was identified as a very potent antileukefic agent. In the present study, effect of natural toxin(plankton) and Kombucha is PSP(GTXI-3, neoSTX), on various MTT assay cervical cancer cell line. Toxin(GTX 1-3, neoSTX) also inhibited the proliferation in primary cervical cancer calls in a dose-dependent toxin concentration. These results showed that toxin was very potent in inhibiting the proliferation of cervical cancer calls in vitro. Toxins and Kombuoha exhibited a dose dependent inhibition of cellular proliferation in cancer cell line.
A bacterial artificial chromosome library of Pectobacterium carotovorum 35 was constructed to characterize the genome and to sequence its hrp region. The hrp cluster of P. carotovorum 35 consisted of 26 open reading frames in five operons. A promoter-based green fluorescent protein technology was used to identify the genes regulated by the alternative sigma factor, HrpL, in P. carotovorum 35. The majority of the selected clones contained the hrpJ operon promoter sequence, which harbors a hrp box, but no putative hrp boxes were detected within the promoter sequences of two other hrpL-regulated genes encoding for pectate lyase and large repetitive protein. Although the promoters of five other hrp operons also contained hrp boxes, their expression was not HrpL-dependent in the promoter-based selection in E. coli. However, transcriptional analysis showed that expression from all operons harboring hrp boxes, except for the hrpN operon, was reduced significantly in the hrpL mutant. The severity of soft-rot symptoms when the hrpL mutant was applied to the surface of tobacco leaves, mimicking natural infection, was greatly attenuated. These results indicate that the hrpL gene of P. carotovorum 35 may be involved in the development of soft-rot symptoms.
A new extended-spectrum ${\beta}-lactamase$ with an isoelectric point (pl) of 6.2 was detected in Klebsiella pneumoniae Fl 61 that was isolated from a patient with infection. This strain was highly resistant to the third or fourth generation cephalosporins such as cceftazidime ceftriaxone, cefoperzaone, and cefpirome. Analysis of this strain by the double disk diffusion test showed synergies between amoxicillin-clavulanate (AMX-CA) and cefotaxime, and AMX-CA and aztreonam, which suggested that this strain produced a extended-spectrum ${\beta}-lactamase$ (ESBL). Cenetic analysis revealed that the resistance was due to the presence of a 9.4-kb plasmic, designated as pkpl 61, encoding for new ${\beta}-lactamase$ gene (bla). Sequence analysis showed that a new bla gene of pkpl 61 differed from $bla_{TEM-1}$ by three mutations leading to the following amino acid substitutions: $Val_{84}{\rightarrow}lie,{\;}Ala_{184}{\rightarrow}Val,{\;}and{\;}Gly_{238}{\rightarrow}Ser$. These mutations have not been reported previously in the TIM type ${\beta}-lactamases$ produced by clinical strains. The novel ${\beta}-lactamase$ was overexpressed in E. coli and purified by ion exchange chromatography on Q-Sepharose and CM-Sepharose, and then further purified by gel filtration on Sehadex G-200. The catalytic activity of th8 purified ${\beta}-lactamase$ was confirmed by the nitrocefin disk.
Yang, Kyoung Hee;Yun, Bohyun;Choi, Hye Jin;Ryu, Sangdon;Lee, Woong Ji;Oh, Mi-Hwa;Song, Min-Ho;Kim, Jong Nam;Oh, Sangnam;Kim, Younghoon;Kim, Young Jun
Food Science of Animal Resources
/
v.39
no.1
/
pp.84-92
/
2019
Listeria monocytogenes is a major cause of serious foodborne illness in the dairy foods. Although Caenorhabditis elegans model is well established as a virulence model of pathogenic bacteria, its application on L. monocytogenes is critically unclear. The objective of this study was to carry out an evaluation of L. monocytogenes toxicity using C. elegans nematode as a simple host model. We found that C. elegans nematodes have high susceptibility to L. monocytogenes infection, as a consequence of accumulation of bacteria in the worms' intestine. However, L. innocua, which is known to be non-toxic, is not accumulate in the intestine of worms and is not toxic similarly to Escherichia coli OP50 known as the normal feed source of C. elegans. Importantly, immune-associated genes of C. elegans were intensely upregulated more than 3.0-fold when they exposed to L. monocytogenes. In conclusion, we established that C. elegans is an effective model for studying the toxicity of L. monocytogenes and we anticipate that this system will result in the discovery of many potential anti-listeria agents for dairy foods.
Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, has been implicated in T7 DNA replication, recombination, and repair. Purified gene 2.5 protein has been shown to interact with the phage encoded gene 5 protein (DNA polymerase) and gene 4 proteins (helicase and primase) and stimulates their activities. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth. T7 phage that contain null mutants of gene 2.5 were constructed by homologous recombination. These mutant phage $(T7\Delta2.5)$ cannot grow in Escherichia coli. After infection of E. coli with $T7\Delta2.5$, host DNA synthesis is shut off, and $T7\Delta2.5$ DNA synthesis is reduced to less than $1\%$ of wild-type phage DNA synthesis (Kim and Richardson, 1993, Proc. Natl. Aca. Sci. USA, 90, 10173-10177). A truncated gene 2.5 protein $(GP2.5-\Delta21C)$ deleted the 21 carboxyl terminal amino acids was constructed by in vitro mutagenesis. $GP2.5-\Delta21C$ cannot substitute for wild-type gene 2.5 protein in vivo; the phage are not viable and exhibit less than $1\%$ of the DNA synthesis observed in wild-type phage-infected cells. $GP2.5-\Delta21C$ has been purified to apparent homogeneity from cells overexpressing its cloned gene. Purified $GP2.5-\Delta21C$ does not physically into「act with T1 gene 4 protein as measured by affinity chromatography and immunoblot analysis. The mutant protein cannot stimulate T7 gene 4 protein activity on RNA-primed DNA synthesis and primer synthesis. These results suggest that C-terminal domain of gene 2.5 protein is essential for protein-protein interactions.
Park, Yong-Woo;Lim, Sun-Teak;Kang, Kyu-Young;Yun, Han-Dae
Applied Biological Chemistry
/
v.38
no.4
/
pp.313-319
/
1995
The involvement of the cell-wall degrading enzymes in Rhizobium has long been an unsolved question about the infection process in the formation of root nodule. To assess the contribution of the cellulase to the nodulation of rhzobia, here we report the production of cellulase from R. meliloti TAL1372 which degrade carboxymethylcellulose (CMC) model substrate with CMC-plate method. We constructed a genomic library by cloning Sau3A-digested genomic DNA from R. meliloti TAL1372 into the BamHI site of the cosmid vector pLAFR3. Out of more than one thousand transductants of E. coli, one clone (pRC8-71) had CM-cellulase activity and contained pLAFR3 cosmid with 30 kb insert of R. meliloti DNA The product of CM-cellulase gene was analyzed by native PAGE. About 45 kD protein was considered to be a product of the gene. Tn5 mutagenesis reveals that the structural gene located in a ca. 3 kb KpnI fragment. The cellulase-minus mutants of R. meliloti TAL1372 were obtained by Tn5 mutagenesis of pRC8-71 and marker exchange techniques. Analyses of the nodulation ability of these Tn5 mutants showed that the CM-cellulase gene of R. meliloti TAL1372 may be involved in early nodulation development on alfalfa (Medicago satiua).
This study was aimed to examine the antibacterial and antioxidative properties of seven edible plants from Thailand to develop alternative antibiotics as feed additives. The plants include Citrus aurantifolia Swingle (Lime) fruits and its leaves, Sesbania grandiflora L. (Agati sesbania) leaves, Piper sarmentosum Roxb (Wild betal) leaves, Curcuma domestica Valeton (Turmeric) roots, Morinda citrifolia L. (Beach mulberry) leaves, Cassia siamea britt (Siamea cassia) leaves, and Cocos nucifera L. (Coconut) peels. The plants were extracted by methanol, n-hexane, chloroform, ethyl acetate, butanol and water. Antibacterial activities with minimum inhibitory concentration (MIC) were determined by agar diffusion assay against Escherichia coli, Burkholderia sp., Haemopilus somnus, Haemopilus parasuis, and Clostridium perfringens that were considered pathogenic strains in livestock infection. Methanol extracts of C. aurantifolia Swingle fruits and leaves showed the broadest spectrum of antibacterial activities except for C. perfringens. Butanol extract of S. grandiflora L. leaves showed the strongest activity against Burkholderia sp. with MIC, $135{\mu}g/mL$. P. sarmentosum Roxb leaves showed antibacterial activities against E. coli, Burkholderia sp. and H. parasuis. Ethyl acetate and water extracts from C. domesitca Valeton roots showed MIC of $306{\mu}g/mL$ and $183{\mu}g/mL$, respectively against only C. perfringens. Antioxidative activity was determined by 2-diphenyl-2-picryl hydrazyl photometric assay. The methanol extracts of C. aurantifolia Swingle fruits and P. sarmentosum Roxb leaves showed the highest antioxidant activity among all the extracts with 3.46 mg/mL and 2.70 mg/mL effective concentration 50% ($EC_{50}$) values, respectively. Total contents of phenolics and flavonoids were measured from the plant extracts. Methanol extracts of S. grandiflora L. and chloroform extracts of C. domestica Valeton were found to have the highest amount of total phenolics, 41.7 and $47.8{\mu}g/mL$, respectively. Flavonoid content of methanol extracts in S. grandiflora L. T was $22.5{\mu}g/mL$ and the highest among plant extracts tested. These results indicated that C. aurantifolia Swingle, S. grandiflora L., P. sarmentosum Roxb, and C. domestica Valeton have antibacterial and antioxidant activities and can be used as alternative antibiotics or potential feed additives for the control of animal pathogenic bacteria.
The antagonistic activities against animal entero-pathogenic bacteria were investigated with 444 natural substances fermented by various probiotics. A white rice product fermented (FWR) by Clostridium butyricum IDCC 9207 with a high growth inhibition of Salmonella typhimurium KCTC 2054 and Escherichia coli O157:H7 was selected. Also, a FWR was shown to suppress 8 among 21 pathogenic bacteria. In a mouse model with salmonella (${\times}10^9$ CFU/mouse) infection, 5 samples (200 ${\mu}{\ell}$/mouse/day) were fed to mice (n = 25) for 18 days. A fermented white rice containing C. butyricum IDCC 9207 (FWRCb9207) among 5 samples significantly inhibited the growth of salmonella, while in the control group (PBS, tetracycline) the number of salmonella increased. And the treatment with FWRCb9207 was found to increase the secretory immunoglobulin A (sIgA) level in the feces of salmonella-infected mice. The results obtained in this study suggest that a FWRCb9207 might be utilized as a feed additive in pigs and poultry diets.
Urinary tract infection (UTI) in children has been known to be a cause of renal damage, leading to scar formation, hypertension and renal failure. And vesico-ureteral reflex (VUR), frequently accompanying UTI in young children, has been incriminated as the main factor causing scar formation. This retrospective study has been undertaken to see the relationship among UTI, VUR and renal scar formation. Study population consisted of 291 children (boy 134, girl 42) with UTI, who have been admitted to the Pediatric Department of Kyungpook University Hospital during 6 1/2 year period from January 1990 to June 1996. VUR was diagnosed by VCUG and renal scar by ultrasonogram, DMSA scan (or DMSA SPECT) and IVP. The following result were obtained. Sexual difference showed male predominance (male to female, 134:42) below 1 year of age, and female predominance (male to female, 11:35) over 5 years of age were rioted. VUR has been found in 64 children (22%) and the degree of reflux, classfied by the method proposed by 'International Reflux Study in Children', were as follows ; Grade I : 4.0%, Grade II : 3.0%, Grade III : 2.7%, Grade IV : 5.8% and Grade V : 6.2%. There was no sexual difference E.coli was the most predominant infecting agent occurring in 167 children (57%), and end-stage renal failure was diagnosed at the time of first admission in 5 children with Grade V VUR. Renal scar has been noted in 49 out of 582 kidneys (8.4%), and the incidence of scar foramation according to the degree of VUR were as follow ; Grade 0 (No reflux) : 1.2%, Grade I : 6.7%, Grade II 27.3%, Grade III 29.4%, Grade IV : 57.1%, and Grade V : 100%. In summary, present study shows that renal scar formation in UTI has close correlation with the severity of VUR occurring more frequently in severe reflux, so that early diagnosis and proper treatment of UTI and VUR is of paramount importance in preventing renal damage in children with UTI.
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