Streptococcus pneumoniae is one of the major pathogens in community-acquired diseases, and it contains several factors that promote its pathogenesis, including pneumolysin (PLY). PLY is a member of the cholesterol-dependent cytolysin family, which attacks cholesterol-containing membranes, thereby forming ring-shaped pores. Thus, it is a major key target for vaccines against pneumococcal disease. We cloned the PLY gene from S. pneumoniae D39 and inserted it into the pQE-30 vector. Recombinant PLY (rPLY) was overexpressed in Escherichia coli M15 and purified by $Ni^{2+}$ affinity chromatography. Similarly, a PLY-EGFP fusion gene was produced by inserting the EGFP gene at the 3' end of the PLY gene in the same vector, and the recombinant protein was purified. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) showed that both recombinant proteins were purified. rPLY exhibited significant hemolytic activity against 1% human red blood cells (RBCs). Complete hemolysis was obtained at 500 ng/ml, and 50% hemolysis was found with a 240 ng/ml concentration. In contrast, rPLY-EGFP did not show hemolytic activity. However, rPLY-EGFP did bind the RBC membrane, indicating that rPLY-EGFP lost hemolytic activity via EGFP fusion, while retaining its membrane-binding ability. These data suggest that PLY's C terminus is important for its hemolytic activity. Therefore, these two recombinant proteins can be extremely useful for investigating the toxin mechanism of PLY and cell damage during pneumonia.
The diagnosis of iron deficiency rests upon the correct evaluation of body iron stores. Morphological interpretation of blood film and the red cell indices are not reliable and often absent in mild iron deficiency. Serum iron levels and iron-binding capacity are more sensitive indices of iron deficiency, but they are often normal in iron depletion and mild iron deficiency anemia. They are also subject to many variables which may introduce substantial errors and influenced by many pathologic and physiologic states. Examination of the bone marrow aspirate for stainable iron has been regarded as one of the most sensitive and reliable diagnostic method for detecting iron deficiency, but this also has limitations. Thus, there is still need for a more practical, but sensitive and reliable substitute as a screening test of iron deficiency. Pollack et al. (1965) observed that the intestinal absorption of cobalt was raised in iron-deficient rats and Valberg et al. (1969) found that cobalt absorption was elevated in patients with iron deficiency. A direct correlation was demonstrated between the amounts of radioiron and radiocobalt absorbed. Unlike iron, excess cobalt was excreted by the kidney, the percentage of radioactivity in the urine being directly related to the percentage absorbed from the gastrointestinal tract. Recently a test based on the urinary excretion of an oral dose of $^{57}Co$ has been proposed as a method for detecting iron deficiency. To assess the diagnostic value of urinary cobalt excretion test cobaltous chloride labelled with $1{\mu}Ci\;of\;^{58}Co$ was given by mouth and the percentage of the test dose excreted in the urine was measured by a gamma counter. The mean 24 hour urinary cobalt excretion in control subjects with normal iron stores was 6.1% ($1.9{\sim}15.2%$). Cobalt excretion was markedly increased in patients with iron deficiency and excreted more than 29% of the dose. In contrast, patients with anemia due to causes other than iron deficiency excreted less than 27%. Hence, 24 hour urinary cobalt excretion of 27% or less in a patient with anemia suggets that the primary cause of the anemia is not iron deficiency. A value greater than 27% in an anemic subject suggests that the anemia is caused by iron deficiency. The cobalt excretion test is a simple, sensitive and accurate method for the assessment of body iron stores. It may be particularly valuable in the epidemiological studies of iron deficiency and repeated evaluations of the body iron stores.
Copper ($CuSO_4$) has been widely used to control algae and pathogens in fish culture ponds. However, its toxic effects on fish depend not only on its concentration in water but also on water quality. The susceptibility of the rockfish, Sebastes schlegeli to copper was evaluated at three water-temperatures (WT; 18, 23 and $28^{\circ}C$) for 4 days. After the exposure of two copper concentrations (100 and $200{\mu}g/L$), a hematological effect was exerted on rockfish, by causing changes in red blood cell count and hematocrit value at $28^{\circ}C$. Total protein levels of the fish showed a tendency of co-increase with glucose depend on the WT, after copper exposure. However, the plasma calcium and magnesium levels were significantly increased at $200{\mu}g/L$ copper, regardless of the WT. Enzymes activities including ALT and LDH in serum were also significantly increased depend upon the copper treatment only. This indicates that inorganic components and enzymes activities were sensitive indexes to stress by toxicant such as copper. The cortisol levels were significantly elevated by both WT rising and copper treatment in serum of rock fish. In conclusion, these changes can be seen as an initial response to temperature stress and as a sustaining response to copper exposure. The present findings suggest that a simultaneous stress by temperature change and copper exposure could accelerate an alteration of hematological and plasma biological parameters in the rockfish.
This study has been performed to investigate the changes in serum lipid and lipoprotein profiles of the national female soccer players during summer training season. Twenty five Korean national soccer players participated in the study. The mean age of the subjects was 23.3$\pm$2.5 years old. Their mean carrier as soccer player was 11.0 $\pm$2.6 years. Mean height was 164.4$\pm$5.7 cm and mean weight was 57.4$\pm$4.6 kg. Mean body fat percent was 23.9$\pm$3.0% and body mass index was 22.4$\pm$6.0. Mean serum triglyc eride concentration was normal (81.0$\pm$26.3 mg $dl^{-1}$), but total cholesterol (224.3$\pm$58.3 mg $dl^{-1}$) and LDL-cholesterol (162.2$\pm$59.0 mg $dl^{-1}$ were higher than normal range. Hematocrit, hemoglobin and total iron binding capacity were low, but in a normal range. Serum triglyceride concentration was negatively correlated with red blood cell counts (r=-0.448, p<0.05). Meanwhile, HDL-cholesterol was positively correlated with hematocrit percent (r=0.408, p<0.05). Therefore, better iron status was related with better serum lipid profiles of the subjects. Four weeks of iron supplementation had no effect on serum lipid and lipoprotein cholesterol of the subjects.
A compact soft x-ray microscope operated in the 'water window' wavelength region ($2.3{\sim}4.4nm$) was used for observing cells with nano-scale spatial resolution. To obtain cellular imaging captured with colloidal gold nanoparticles using a compact soft x-ray microscope. The colloidal gold nanoparticles showed higher contrast and lower transmission more than 7 times than that of cellular protein on the soft x-ray wavelength region. The structure and thickness of the cell membrane of the Coscinodiscus oculoides (diatome) and red blood cells were seen clearly. The gold nanoparticles within the HT1080 and MDA-MB 231 cells were seen clearly on the soft x-ray microscopy. The gold nanoparticles were aggregated within vesicles by endocytosis.
Kim, Suhee;Yu, Do-Hyeon;Kang, Sung-Woo;Chae, Jeong-Byoung;Choi, Kyoung-Seong;Kim, Hyeon-Cheol;Park, Bae-Keun;Chae, Joon-Seok;Park, Jinho
Parasites, Hosts and Diseases
/
v.55
no.5
/
pp.481-489
/
2017
Tick-borne pathogens can cause serious problems in grazing cattle. However, little information is available on tick-mediated diseases in cattle grazing on mountains. Thus, this study aimed to understand the potential problems related to tick-borne diseases in grazing cattle through the investigation of prevalent tick-transmitted infections, and their associated hematological changes, in terms of season and grazing type in Korean indigenous cattle (=Hanwoo). Hanwoo cattle from 3 regions of the Republic of Korea (=Korea) were either maintained indoors or placed on grassy mountains from spring to fall of 2014 and 2015. Cattle that grazed in mountainous areas showed a greater prevalence of tick-borne infections with an increased Theileria orientalis infection rate (54.7%) compared to that in non-grazing cattle (16.3%) (P<0.001). Accordingly, the red blood cell (RBC) count and hematocrit (HCT) values of grazing cattle were significantly lower than those of non-grazing cattle throughout the season (P<0.05). Moreover, RBC, hemoglobin (Hb), and HCT of T. orientalis-positive group were significantly lower than those of T. orientalis-negative group (P<0.05). T. orientalis is a widespread tick-borne pathogen in Korea. Grazing of cattle in mountainous areas is closely associated with an increase in T. orientalis infection (RR=3.4, P<0.001), and with consequent decreases in RBC count and HCT. Thus, these findings suggest that the Hanwoo cattle in mountainous areas of Korea are at a high risk of infection by T. orientalis, which can lead to hematological alterations. This study highlights the necessity of preventive strategies that target T. orientalis infection.
One hundred and sixty Taiwan country chickens (d-old chicks) were randomly assigned into four groups with four replicates and equal sex. Basal diets were supplemented with 0, 0.5, 1 and 2% of Bazhen powder, a traditional Chinese herbal medicine complex. The study was conducted for 14 wks. Experimental results indicated that Bazhen supplement did not influence feed intake, body weight gain and feed:gain ratio. Compared with control group, the percentage of serum HDL (high-density lipoprotein) linearly increased (p<0.03) and that of VLDL+LDL (very low-density+low-density lipoprotein) linearly decreased (p<0.03) in Bazhen supplemented groups, that 2% Bazhen was significantly different with control group (p<0.05). Chickens fed diets containing 2% Bazhen displayed reduced (p<0.05) serum GOT (glutamic-oxaloacetic transaminase) levels. The IgG, ${\gamma}$-globulin levels and PHA (phytohemagglutinin) skin challenge results in 1% Bazhan supplemented group were higher (p<0.05) than in the control group, the SRBC (sheep red blood cell) and ND (newcastle disease) titers in Bazhen supplemented groups were linear higher (p<0.05) than in the control group. The liver catalase activity and the capacity of scavenging DPPH (${\alpha}$-${\alpha}$-diphenyl-${\beta}$-picrylhydrazyl) radical were linearly increased (p<0.03) in Bazhen supplemented groups, and the 1 and 2% groups were different from the control group (p<0.05). Liver TBARS (thiobarbituric acid-reactive substances) levels in all Bazhen supplemented groups and total glutathione level in the 2% group were reduced (p<0.05) compared to the control group and displayed a linear response (p<0.05). The TBA (thiobarbituric acid) and pH value of the breast muscle after 24 h post-mortem in the Bazhen supplemented groups was linear lower (p<0.05) than in the control group. Results from this study demonstrated that Bazhen supplement in chicken had several beneficial effects, including increased SRBC and ND titers, HDL and IgG, ${\gamma}$-globulin levels, PHA skin challenge result, decreased VLDL+LDL and GOT levels, and displayed antioxidation effects in serum and carcass meat parameters.
Objective: This study aimed to examine the effect of sodium butyrate (SB) on growth performance, immune status, organs weights, and microarchitecture of lymphoid organs and small intestine. Methods: A total of 120, 1-d-old broiler chicks were distributed into the following four treatment groups: corn-soy based basal diet (BD) without supplement (control), or the same BD supplemented with 0.1 g/kg zinc bacitracin (ZnB), 0.5 g/kg SB (SB-0.5), or 1.0 g/kg SB (SB-1), respectively. Six birds/group were killed on d-21 and d-35, and samples were collected. Results: Cell-mediated immune response at 48 h post-Phytohemagglutinin-P injection, and antibody titer against Newcastle disease vaccine and sheep red blood cells on d-35 was noted higher (p<0.05) in SB-1 compared to ZnB and control. Lower (p<0.05) feed conversion ratio (FCR) was attained by the supplemented groups. Thymus and spleen weighed more (p<0.05) in SB-1, and bursa registered more (p<0.05) weight in both SB groups compared to control. On d-21, areas of thymus medulla and spleen germinal centers were noted higher (p<0.05) in SB-1 group. The villus height and villus surface area increased (p<0.05) in duodenum and jejunum in both SB groups on d-21, and in SB-1 on d-35, respectively compared to ZnB and control. On d-21, number of goblet cells containing mucins of acidic nature increased (p<0.05) in all the segments of small intestines in SB-1 group compared to control, and on d-35 in ileum compared to other groups. Conclusion: In conclusion, SB improved growth performance and immunity as well as modulated morphology of lymphoid organs and gut mucosa in broiler chickens.
Purpose: This study was to determine the efficacy and safety of pemetrexed based chemotherapy in treating patients with metastatic gastric cancer who failed to respond to first and (or) second line chemotherapy. Patients and Methods: Metastatic gastric cancer patients who failed first and (or) second line chemotherapy, were enrolled. All patients were recruited from Jiangsu Cancer Hospital & Research Institute, and were treated with pemetrexed $500mg/m2$ (intravenous; on day 1), and a platinum (or irinotecan) every 3 weeks until disease progression, or intolerable toxicity. Evaluation on efficacy was conducted after two cycles of chemotherapy using the Response Evaluation Criteria for Solid Tumors. Toxicity was recorded according to National Cancer Institute Common Terminology Criteria for Adverse Events version 3.0. Results: From Jun 2011 to May 2013, 23 patients were enrolled. All eligible 23 patients completed at least 2 cycles of chemotherapy with pemetrexed based chemotherapy, and were evaluable. Their median age was 55 years (range 40 to 78 years). Seventeen patients were male and 6 female. Three patients (13%) achieved partial response, five patients (22%) stable, 15 patients (65%) with disease progression, and none with complete response. Grade 2 neutrophil suppression occurred in 4.3%, grade 3 in 13% of patients, and no grade 4 was reported. Thrombocytopenia was encountered as follows: 4.3% grade 2, 4.3% grade 3 and 4.3% grade 4. Incidence of anemia was 34.8% in grade 2, 8.7% grade 3 and 0% grade 4. Only 4.3% of patients required packed red blood cell infusion. Elevated transaminase were 4.3% in grade 2 and 0% in grade 3 or 4. Other toxicity included oral mucositis. Conclusions: Pemetrexed based chemotherapy is mildly effective in treating patients with metastatic gastric cancer with tolerable toxicity.
The effects of dried leaf powders and water and ethanol extracts of persimmon and green tea on lipid metabolism, lipid peroxidation and antioxidative enzyme activity were investigated in 12-month-old rats. Forty-nine male Sprague-Dawley rats weighing 520$\pm$19g were blocked into seven groups according to body weight. Rats were raised for four weeks with control(no tea leaf powder or extracts) and experimental diets containing either 5%(w/w) dried leaf powders of persimmon(Diospyros kaki Thunb) or green tea(Camellia sinensis O. Ktze), or water or ethanol extract from equal amounts of each dried tea powder. Food intakes of all tea diet groups were higher than that of control. Weight gains and food efficiency ratios of all tea diet groups were not significantly different from those of control. All tea diets decreased plasma triglyceride level, especially, green tea powder and persimmon ethanol diets were more effective than other diet. All the tea diet groups showed decrease in liver triglyceride level, and persimmon powder and ethanol extract increased fecal triglyceride excretion. Plasma cholesterol levels of all the tea diet groups were not significantly different from the control, but control. Fecal cholesterliver cholesterol concegroups were significantlntrations of all tea y lower than that of ol excretions of persimmon powder, green tea ethanol extract, persommon ethanol extract and green tea ethanol extract groups were significantly higher than that of control. Plasma and liver thiobarbituric acid reactive substance(TBARS) concentrations of all the tea diet groups were lower than that of control. Especially, plasma TBARS concentrations of green tea powder and persimmon ethanol extract groups were sinificantly low. Red blood cell(RBC) superoxide dismutase(SOD) activities of persimmon ethanol extract and green tea water extract groups were increased, and RBC catalase activities of all experimental groups were not significantly different. RBC glutathione peroxidase(GSH-px) activities of persimmon ethanol extract, persimmon water extract and green tea powder groups were increased. Liver SOD activities of all the tea diet groups except green tea ethanol extract group were higher than that of control. Liver catalase activities of all experimental groups were not significantly different, and liver GSH-px activity of green tea powder group was significantly higher than that of control. In conclusion, dried leaf powders, and water and ethanol extracts of persimmon and green tea were effective in lowering lipid level, inhibiting lipid peroxidation and increasing antioxdative enzyme activities in 12-month-old rat. Green tea leaf powder with high contents of flavonoids and water soluble dietar fiber was most effective in lowering plasma triglyceride, cholesterol and TBARS level. (Korean J Nutrition 34(3) : 285~298, 2001)
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