The giant mealworm beetle, Zophobas atratus (Coleoptera: Tenebrionidae) has been used as a protein source for small pets and mammals. Recently, it was temporarily registered in the list of the Food Code. We previously performed an in silico analysis of the Zophobas atratus transcriptome to identify putative antimicrobial peptides and identified several antimicrobial peptide candidates. Among them, we assessed the antimicrobial and anti-inflammatory activities of zophobacin 1 that was selected bio-informatically based on its physicochemical properties against microorganisms and mouse macrophage Raw264.7 cells. Zophobacin 1 showed antimicrobial activities against microorganisms without inducing hemolysis and decreased the nitric oxide production of the lipopolysaccharide-induced Raw264.7 cells. Moreover, ELISA and Western blot analysis revealed that zophobacin 1 reduced expression levels of pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). We also investigated expression of pro-inflammatory cytokines (interleukin-6 and interleukin-1β) production through quantitative real time-PCR and ELISA. Zophobacin 1 markedly reduced the expression level of cytokines through the regulation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) signaling. We confirmed that zophobacin 1 bound to bacterial cell membranes via a specific interaction with lipopolysaccharides. These data suggest that zophobacin 1 could be promising molecules for development as antimicrobial and anti-inflammatory therapeutic agents.
In this article, we investigate into the structural changes of liposome to design its functional membranes by the synthesis of two types of liposomes, DPPC liposome and DTAB of hydrocarbon substance/DPPC liposome. The changes of membrane structures are evaluated by the CF fluorescent intensity measured above and below the phase transition temperature of the membrane, $t_c=41^{\circ}C$. CF fluorescent intensities are enhanced by the CF leakage from DPPC liposome at $45^{\circ}C$, while no changes are observed at $20^{\circ}C$. Under the same conditions, it is observed that the intensity enhanced by CF leakage from DPPC/DTAB liposome is larger than that of DPPC liposome alone, which suggests that DPPC/DTAB liposome has irregular arrangement. Under the presence of $Ca^{2+}$, Quin 2 fluorescent intensity in either DPPC liposome or DPPC/DTAB liposome is significantly increasing at $45^{\circ}C$, while almost none of the changes are observed at $20^{\circ}C$. The fluorescent intensity of DPPC liposome turns out to be larger than that of DPPC/DTAB liposome, which suggests that the DPPC/DTAB liposome is structurally more stable than the DPPC liposome. Additionally, when the analysis is done to observe changes in the shapes of membrane surfaces with ANS fluorescent, ANS fluorescent under DPPC or DPPC/DTAB liposome shows each of different appearances at $45^{\circ}C$ and $20^{\circ}C$ respectively. This result indicates that its respective membrane fluidity is changing above and below of the designated temperatures in phase transition. As to the magnitude of change of its membrane fluidity, DPPC liposome is much larger than DPPC/DTAB liposome. As far as the temperature in phase transition measured by DSC are concerned, it is $41^{\circ}C$ and $32^{\circ}C$ for DPPC and DPPC/DTAB liposome respectively, which suggests that DPPC/DTAB liposome has an irregular molecular arrangement in its structure. That is, it is summed up that DPPC/DTAB turns out to be structurally stable, even so, its structure is irregularly arranged.
Purpose : Thymus size can be affected by several factors and perinatal diseases can be estimated by its size. The purpose of this study was to search for a relationship between cardiothymic/thoracic(CT/T) ratio and perinatal diseases such as neonatal respiratory distress syndrome(RDS) and intrauterine growth retardation(IUGR) by measuring the width of the cardiothymic shadow at the level of the carina and dividing it by the width of the thorax at the costophrenic angles. Methods : A clinical study was conducted on newborn infants with RDS(n=51), IUGR(n=27), and premature rupture of membranes(PROM, n=48), who were admitted at NICU of Catholic University of Daegu from June 2000 to Oct. 2001. CT/T ratio was measured within six hrs of age, at 2-3 days of age, and at 5-7 days of age. Results : CT/T ratios of RDS group, IUGR group, and PROM group were $0.46{\pm}0.07$, $0.32{\pm}0.04$. $0.36{\pm}0.06$, respectively. CT/T ratios of RDS group within 6hrs of age, at 2-3 days of age, and at 5-7 days of age were $0.43{\pm}0.07$, $0.34{\pm}0.06$, $0.25{\pm}0.04$, respectively. There were statistically significances among the RDS group, the IUGR group, and the PROM group and in the RDS group at different times. Regression for gestational age among three groups was not statistically significant but correlation for gestational age in the entire groups was statistically significant. CT/T ratio between normal spontaneous vaginal delivery and c-section among three groups was not statistically significant. CT/T ratios with dexamethasone-treated group and untreated group was not statistically significant. Conclusion : We concluded that thymus size differed significantly in the perinatal diseases such as RDS and IUGR, and so can be used as an early diagnostic tool for perinatal diseases.
This study was designed to investigate the effects of a butanol (BuOH) fraction from an extract of pine (Pinus densiflora Sieb et Zucc.) needles, on oxygen radicals and their scavenger enzymes in the liver membranes of rats. Twenty-eight male Sprague-Dawley (SD) rats were divided into four groups over a 45 days study period: the control group on a basic diet, and three experimental groups on three different dietary levels of the butanol fraction, specifically 25 mg (BuOH-25), 50 mg (BuOH-50), and 100 mg (BuOH-100) butanol fraction/kg body weight/day, thereby 0.025%, 0.05%, 0.1% of butanol extract of pine needles was added to basil diet respectively. At the end of the experimental period, body weights and food intakes were not different among the four groups. The results showed that cholesterol accumulation in the mitochondria and microsomes of liver cells was significantly inhibited in the BuOH-50 and BuOH-100 groups: by 11.6% and 20.1% in the mitochondria of the BuOH-50 and BuOH-100 groups, respectively; and by 10.5%, and 13.5% in the microsomes of the BuOH-50 and BuOH-100 groups, respectively, compared with the control group. The levels of hydroxyl radicals (.OH) were significantly) lower in the liver mitochondria of the BuOH-50 and BuOH-100 groups (by 13.3% and 18.5%, respectively), while OH radicals were significantly lower in the microsomes or all three experimental groups (by 15.7% in the BuOH-25 group, 20.0% in the BuOH-50 group, and 20.6% in the BuOH-100group), compared with the control group. Superoxide radical (O$_2$) formation was also significantly inhibited in the liver cytosol of both BuOH-50 and BuOH-100 groups; the levels of these radicals were 8.0% lower for the BuOH-50 group and 11.1% lower for the BuOH-100 group, compared to the control group. Copper/Zinc - superoxide dismutase (Cu/Zn-SOD) activities were significantly increased (by 10.3% and 15.9%, respectively) in the liver cytosols of the BuOH-50 and BuOH-100 groups, but Mn-SOD activities were almost identical in the three RuOH groups, compared with the control group. Glutathione peroxidase (GPx) activities were significantly increased in the three experimental groups (by 9.0% in the BuOH-25 group, 19.4% in the BuOH-50 group, and by 25.6% in the BuOH-100 group), compared with the control group. These results suggest that the butanol extract of pine needles may play an effective role in attenuating oxygen radicals and activating scavenger enzymes; consequently, aging may be very effectively modulated and/or inhibited.
The purpose of this study was to determine the effect of energy supplement on responses of plasma insulin-like growth factor (IGF)-1 and IGF binding proteins (IGFBPs) to growth hormone-releasing peptide-2 (GHRP-2) administration in normal protein-fed wethers, and to observe the effect of GHRP-2 treatment on hepatic growth hormone (GH) receptor in well-fed wethers. Plasma IGF-1 and 39-42 kDa IGFBP-3 during the HENP (CP, crude protein 0.34 and TDN, total digestible nutrients 1.83 kg/day DM, dry matter intake) treatment period were higher than in the LENP (CP 0.32 kg and TDN 0.87 kg/day DM intake) period (P<0.05). The response of GH was stimulated by GHRP-2 ($12.5\;{\mu}g/kg$ body weight/day) administration during both of the feed treatment periods (P<0.05). The area under curve (AUC) increment and average concentration of GH (0-180 min) with GHRP-2 administration was higher during HENP treatment than LENP treatment (P<0.01). During the HENP treatment period from day 1 to day 7 of twice daily GHRP-2 treatment, the plasma IGF-1 increment was increased on days 2, 6 and 7 of GHRP-2 administration (P<0.05). On the basis of ligand blotting, the proportions of plasma 39-43 kDa IGFBP-3 during the HENP treatment period only showed a significant difference on days 6 and 7 with GHRP-2 administration. No significant difference in the specific binding of $^{125}I-labeled$ oGH to hepatic membranes was detected between the saline and GHRP-2 treatments of the HENP-fed wethers. These results suggest that the nutritional balance between energy and protein may affect the endogenous GH / IGF-1 axis as well as plasma IGFBP-3 levels.
The goal of periodontal treatment is regeneration of the periodontium. Bone graft and absorbable PLA/PGA membrane have been used for this purpose. In this study, 4${\times}$4mm 1-wall intrabony defects were surgically created bilaterally in the mandible of five male beagles. The control group went through a conventional flap operation, while the experimental group I was treated with absorbable PLA/PGA membranes only, group II was treated with absorbable membrane and calcium phosphate. The results are the following : 1. The defect height was 4.82${\pm}$0.45mm in the control group, 4.93${\pm}$0.79mm in the experimental I group, and 4.92${\pm}$0.62mm in the experimental II group. There was no statistically significant difference among 3 groups(P <0.05). 2. The amount of junctional epithelium migration was 30.90${\pm}$9.92% of the defect height in the control group, 39.16${\pm}$7.51% in the experimental I group, and 38.68${\pm}$12.22% in the experimental II group. There was no statistically significant difference among 3 groups(P <0.05). 3. The amount of connective tissue adhesion was 36.38${\pm}$9.03% in the control group, 14.73${\pm}$3.93% in experimental I group, and 27.87${\pm}$9.70% experimental II group. Experimental group I was a statistically significantly different from control group(P <0.05). 4. The amount of new cementum regeneration was 32.92${\pm}$10.51%, 50.04${\pm}$7.61%, and 39.62${\pm}$12.14% for the control, experimental I, and experimental II group respectively. Experimental group I was a statistically significantly different from control group(P<0.05). 5. The amount of new alveolar bone regeneration was 27.24${\pm}$7.49%, 40.75${\pm}$8.03%, and 36.47${\pm}$15.11% for the control, experimental I, and experimental II group respectively. Experimental group I was a statistically significantly different from control group(P <0.05). The results suggest that the use of PLA/PGA membrane in 1-wall intrabony defect of beagle dogs may promote periodontal regeneration. Further studies are required to determine their regeneration effects.
Journal of the Korean Society of Food Science and Nutrition
/
v.32
no.4
/
pp.506-512
/
2003
Peach wine was fermented at $25^{\circ}C$ for 2 weeks using Saccharomyces cerevisiae KCCM 12224, aged at 15$^{\circ}C$ for 14 weeks, and its physicochemical and microbiological changes were investigated. The viable bacterial cell numbers, 1.4$\times$10$^3$ CFU/mL at the beginning of fermentation, increased to 2.8$\times$10$^{6}$ CFU/mL after 2 weeks, but decreased to 7.0$\times$10$^3$ CFU/mL after 14 weeks. The viable yeast cell numbers were changed from 3.4$\times$10$^2$ CFU/mL to 2.4$\times$10$^{7}$ CFU/mL during fermentation, and decreased to 4.0$\times$10$^4$ CFU/mL after aging. Turbidity total sugar content, reducing sugar content, solid content and b value of peach wine decreased during fermentation but acidity, alcohol content, L and a value increased. Most physicochemical properties except alcohol content and reducing sugar content were not changed significantly during aging. When peach wine was filtered through 0.45 ${\mu}{\textrm}{m}$ nitrocellulose membrane followed by various ultrafiltration membranes with different molecular weight cut-off values, Biomax 100K membrane, with 79 liter/$m^2$/h (LMH) of initial flux, was suitable for ultrafiltration process of peach wine. These membrane filtration treatments resulted in complete removal of microorganisms and decrease in turbidity and alcohol content without changes in other chemical properties. The physicochemical properties of peach wine were not changed and any microorganisms were not found during the storage at 3$0^{\circ}C$ for 12 Weeks.
The lipids of milk provide energy and many essential nutrients for the newborn animal. They also have distinctive physical properties that affect the processing of dairy products. Milk fat globules mainly consist of neutral lipids like triacylglycerols, whereas the globule membranes contain the complex lipids mostly, Phospholipids are a small but important fraction of the milk lipids and are found mainly in the milk fat globule membrane and other membranous material in the skim-milk phase. The milk fats of ruminant animals are characterized by the presence of relatively high concentrations of short-chain fatty acids, especially butyric and hexanoic acids, which are rarely found in milks of non-ruminants. The fatty acids of milk lipids arise from de novo synthesis in the mammary gland and uptake from the circulating blood. The fatty acid compositions of milks are usually complex and distinctive, depending on the nature of the fatty acids synthesized de novo in the mammary gland and those received from the diet in each species. The content and composition of milks from different species vary widely; presumably, these are evolutionary adaptations to differing environments. The actual process by which these globules are formed is unkonwn, but there are indications that triglyceride-containing vesicles which bleb from endoplasmic reticulum may serve as nucleation sites for globules. Recent studies on milk have centred on the manipulation of milk lipids to increase specific fatty acids, i.e. 20-carbon omega-3 fatty acids (eicosapentaenoic acid 20:5n3, decosahexaenoic acid 22:6n3) from marine sources because the fatty acids are closely associated with a decreased risk of coronary heart disease.
Streptococcus mutans (S. mutans), which plays a major role in the etiology of human dental caries, is able to tolerate exposure to acid shock in addition to its acidogenicity. We investigated the effects of pH stress on membrane fluidity, activities and expression levels of F-ATPase, and proton permeability in S. mutans. Using 1,6-diphenyl-1,3,5-hexatriene, we observed membrane ordering at pH 4.8 and pH 8.8. The ordering effects were larger at pH 4.8 in cytoplasmic membranes isolated from S. mutans (CMSM). Increasing pH resulted in a decrease in the activities and expression levels of F-ATPase. The proton permeability was decreased at both acidic and alkaline pHs, and the lowest permeability was observed at pH 4.8. The lower permeability at pH 8.8 than pH 6.8 is likely to be caused by the decreased proton influx due to the decreased CMSM fluidity. In addition, it seems to be evident that extremely low permeability at pH 4.8 was caused by the decreased proton influx due to the decreased CMSM fluidity as well as the increased proton efflux due to the increased activity and expression level of F-ATPase. It is likely that CMSM fluidity and F-ATPase activity are two major key factors that determine proton permeability in S. mutans. We suggest that CMSM fluidity plays an important role in the determination of proton permeability, which sheds light on the possibility of using nonspecific membrane fluidizers, e.g., ethanol, for anti-caries purposes.
Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.
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