Background: Brucella infection induces brucellosis, a zoonotic disease. The intracellular circulation process and virulence of Brucella mainly depend on its type IV secretion system (T4SS) expressing secretory effectors. Secreted protein BspJ is a nucleomodulin of Brucella that invades the host cell nucleus. BspJ mediates host energy synthesis and apoptosis through interaction with proteins. However, the mechanism of BspJ as it affects the intracellular survival of Brucella remains to be clarified. Objectives: To verify the functions of nucleomodulin BspJ in Brucella's intracellular infection cycles. Methods: Constructed Brucella abortus BspJ gene deletion strain (B. abortus ∆BspJ) and complement strain (B. abortus pBspJ) and studied their roles in the proliferation of Brucella both in vivo and in vitro. Results: BspJ gene deletion reduced the survival and intracellular proliferation of Brucella at the replicating Brucella-containing vacuoles (rBCV) stage. Compared with the parent strain, the colonization ability of the bacteria in mice was significantly reduced, causing less inflammatory infiltration and pathological damage. We also found that the knockout of BspJ altered the secretion of cytokines (interleukin [IL]-6, IL-1β, IL-10, tumor necrosis factor-α, interferon-γ) in host cells and in mice to affect the intracellular survival of Brucella. Conclusions: BspJ is extremely important for the circulatory proliferation of Brucella in the host, and it may be involved in a previously unknown mechanism of Brucella's intracellular survival.
ATP in quantity co-stored with neurotransmitters in the secretory vesicles of neurons, by being co-released with the neurotransmitters, takes an important role to modulate the stimulus-secretion response of neurotransmitters. Here, in this study, the modulatory effect of ATP was studied in $Ca^{2+}$ channels of cultured rat adrenal chromaffin cells to investigate the physiological role of ATP in neurons. The $Ca^{2+}$ channel current was recorded in a whole-cell patch clamp configuration, which was modulated by ATP. In 10 mM $Ba^{2+}$ bath solution, ATP treatment (0.1 mM) decreased the $Ba^{2+}$ current by an average of $36{\pm}6%$ (n=8), showing a dose-dependency within the range of $10^{-4}{\sim}10^{-1}mM$. The current was recovered by ATP washout, demonstrating its reversible pattern. This current blockade effect of ATP was disinhibited by a large prepulse up to +80 mV, since the $Ba^{2+}$ current increment was larger when treated with ATP ($37{\pm}5%$, n=11) compared to the control ($25{\pm}3%$, n=12, without ATP). The $Ba^{2+}$ current was recorded with $GTP{\gamma}S$, the non-hydrolyzable GTP analogue, to determine if the blocking effect of ATP was mediated by G-protein. The $Ba^{2+}$ current decreased down to 45% of control with $GTP{\gamma}S$. With a large prepulse (+80 mV), the current increment was $34{\pm}4%$ (n=19), which $25{\pm}3%$ (n=12) under control condition (without $GTP{\gamma}S$). The $Ba^{2+}$ current waveform was well fitted to a single-exponential curve for the control, while a double-exponential curve best fitted the current signal with ATP or $GTP{\gamma}S$. In other words, a slow activation component appeared with ATP or $GTP{\gamma}S$, which suggested that both ATP and $GTP{\gamma}S$ caused slower activation of $Ca^{2+}$ channels via the same mechanism. The results suggest that ATP may block the $Ca^{2+}$ channels by G-protein and this $Ca^{2+}$ channel blocking effect of ATP is important in autocrine (or paracrine) inhibition of adrenaline secretion in chromaffin cell.
Ultrastructural changes in the cells of the corpora allata of the pine moth, Dendrolimus spectabilis Butler, were studied by electron microscope to know the structural correlates of hormone production by the gland during the larval-pupal-adult transformations. Mitochondria are in active phases from the overwintered to the last instar larvae and from the pupae just after pupation to the 20-day old pupae, while they are in inactive phases from the making cocoon stage to the prepupae just before pupation. The peripheral allatum cells have electron dense granules in the intracellular vacuoles of smooth-surfaced endoplasmic reticulum in the larval life, particularly in the overwintered larvae and in the early adults but the swollen smooth-surfaced intracytoplasmic vacuoles made by expansion of an end of the tubular rough endoplasmic reticulum, some of which contain fibrous proteins, are observed in addition to the vacuoles in the intercellular spaces in which the vacuoles grow by fusing each other from the mature larvae to the prepupae, both of them disappearing during just before pupation. After pupation the cytolasmic vacuoles develop again in the allatum cells so that they seem to begin the secretory activity. The fact that the neurosecretory granules stored within the axons terminated in the corpus allatum are visible only from the 20-day old pupa about two days before abult emergence to the 5-day old adult means that the secretion from the allatum cells is under the control of the brain from the late pupal stage, while the secretion during from the larval to the early pupal life has no relation with the brain, because such granules are not observed within the axons. It is, therefore, suggested that at least two kinds of hormone are released with the ages as far as concerned with the production and secretion mechanisms of the allatum hormone: juvenile hormone is released until the last instar larvae without any direct stimlation of the brain and gonadotropic hormone is secreted from the late pupa to the adult by getting brain's stimulation and that the secretory phases observed from the mature larvae to prepupae are presumably concerned with the biosynthesis of protein owing to the ecdysone and those from the early pupal stage in uncontrolled condition of the brain with the prothoracotropic activity.
The study was conducted to find out the concentrations of sex hormones and the contents of metabolites in serum, and the changes of weights and tissues on ovary, thyroid gland and adrenal gland according to gestation period in rabbit. The results were summarized as follows: 1. The ovary weights were increased significantly with the time e lapse after gestation and recovered normally at 5 days after parturition. In the histological changes of ovary, the lutein cells were hypertrophied and the secretory granules were increased actively until 3 weeks after gestation, and then a trophied thereafter. 2. The thyroid gland weights at all observation times were higher than those in control group, and the significance was recognized at 1, 3 and 4 weeks after gestation. The histological features of the secretory epithelium were the hyper trophic and columnar condition stimulating the functional state from 1 week after gestation. 3. The adrenal gland weights in experimental group were recognized significantly at 4 weeks after gestation, but showed higher than those in control group at all observation times. The zona fasciculata and zona reticularis of the gland showed the slight hypertrophic condition, but the zona glomeerulosa and adrenal medulla did not find out any particular changes. 4. The serum concentrations of progesterone and LH reached a peak level at 2 weeks and 1 week after geestation respectively, and rapidly began to decline thereafter. 5. The serum concentrations of estradiol-$17{\beta}$ and FSH were not detected below 20.0 pg/ml and 1.3 mIU/ml respectively. 6. The contents of total protein and non-protein nitrogen nitrogen were decreased gradually with the time elapse after gestation, but the significant differences were recognized from 3 weeks. 7. The total lipids were not changed markedly until 3 weeks, but increased significantly at 4 weeks after gestation and at 5 days after parturition. 8. The serum cholesterol tended to be decreased until 3 weeks, but increased at 4 weeks after gestation and at 5 days after parturition. 9. The serum calcium showed a continuous decrease during the gestation period, but the significant differences were recognized at 3 and 4 weeks. The serum phosphorus also had a significant decrease at 4 weeks after gestation.
Park, J.A.;Kang, H.K.;Chae, E.J.;Seo, K.S.;Kim, S.H.;Yun, C.H.;Moon, Y.S.
Journal of Animal Science and Technology
/
v.49
no.5
/
pp.577-584
/
2007
Adipocyte-specific secretory factor(ADSF)/resistin, an hormone, is a small cysteine-rich protein secreted from adipose tissue and ADSF/resistin has been implicated in modulating adipogenesis in human and rodents. Although the exact role of ADSF/resistin in bovine has not been identified, it may have directly or indirectly involved in adipocyte differentiation. The objective of this study was to investigate its DNA polymorphism associated with carcass traits in Korean Native Cattle(Hanwoo). To investigate DNA polymorphism in Hanwoo ADSF/resistin gene, blood samples were taken from 295 Hanwoo steers belonging to progeny testing at Hanwoo Improvement Center in Korea. Seven single nucleotide polymorphisms(SNPs) were found in intron regions but not in any other regions including promoter (1.7kb) and 4 exons. The highest frequency among SNPs was C186A(0.16/0.84) following G964A (0.156/0.884). The significant correlation(P<0.05) between the SNPs and economic traits was found on 764Ains associated with marbling but not from any other SNPs determined. A computer simulation was also conducted to assess the efficiency of marker assisted selection(MAS) versus the conventional breeding scheme. Results revealed that MAS was more efficient as a breeding tool compared to the conventional. In conclusion, ADSF/Resistin gene is one of candidate genes to evaluate the quality, especially marbling score, in Hanwoo.
In order to observe the antigenic localization in the tissues of Metngonimus yokogawai in growth stages, immunogoldlabeling method was applied to using serum of the cat which Infected with isolated metacercariae from Plecoglossus aztivelis. The sectioned worm tissues from each growth stages were embedded in Lowicryl HM 20 medium, stained with infected serum IgG and protein A gold complect (particle size: 12 nm) and observed by electron microscopy. In the worm tissues of all experimental groups, the geld particles were specifically concentrated on the tegumental synch- tium and cytoplasm of the tegumental cell as well as the secretory granules in the parenchymal tissue. In the 16th and 20th week grown worm tissues, the gold particles were specifically concentrated on the vesicles in the tegumental syncytium and cl·toplasm of the tegumental cell. The gold particles were specifically concentrated on the caecal epithelia of the 4th, 8th and 12th week growth groups but slightly concentrated on those of the 16th and 20th week.
This study was designed to investigate the effects of basal diets supplemented with a clay product consisting of zeolite and attapulgite (ZA) at 1:1 ratio on growth performance, digestibility of feed nutrients, activities of digestive enzymes in small intestine and intestinal health in broiler chickens. In experiment 1, 112 one-day-old male chickens were randomly divided into 2 groups with 8 replicates of 7 chickens each. In experiment 2, 84 one-day-old male chickens were randomly allocated into 2 groups consisting 6 replicates of 7 chickens each. The experimental diets both consisted of a maize-soybean basal control diet supplemented with 0% or 2% ZA. The diets were fed from 1 to 42 days of age. The results showed that ZA supplementation could increase body weight gain (BWG) and feed intake (FI), but had no significant effect on feed conversion ratio. The apparent digestibility values of crude protein and gross energy were significantly increased (p<0.05) by ZA from 14 to 16 d and 35 to 37 d. Dietary ZA treatment significantly increased (p<0.05) the activities of amylase, lipase and trypsin in jejunal digesta and the activities of maltase and sucrase in jejunal mucosa on days 21 and 42. The ZA supplementation also significantly increased (p<0.05) the catalase activity, reduced (p<0.05) the malondialdehyde concentration in the jejunal mucosa. In addition, a decrease of serum diamine oxidase activity and an increase (p<0.05) in concentration of secretory immunoglobulin A in jejunal mucosa were observed in birds treated with ZA on 21 and 42 days. It is concluded that ZA supplementation (2%) could partially improve the growth performance by increasing BWG and FI. This improvement was achieved through increasing the secretion of digestive enzymes, enhancing the digestibilites of nutrients, promoting intestinal health of broiler chickens.
Kim, Myung-Suk;Yoon, Shin-Hee;Hahn, Sang-June;Kim, Mie-Hye
The Korean Journal of Physiology
/
v.22
no.2
/
pp.273-280
/
1988
This study was undertaken to investigate the effect of the medial amygdaloid nucleus on the pancreatic exocrine secretion and plasma secretin concentration in 44 male albino rats. Twenty-three rats in which the medial amygdaloid nucleus was damaged bilaterally by radio frequency a.c. through stereotaxically inserted electrodes (medical amygdaloid group, MA) and twenty-one rats which received the same operation without damage (operated control, OC), were prepared. Under urethan anesthesia, 0.01 N hydrochloric acid (HCl) or physiological saline (0.9% NaCl) was infused at a rate of 0.18 ml/min into the duodenum for 20 minutes. Pancreatic jucie was collected for the 20 min infusion period. After collection of pancreatic juice, blood was sampled from the abdominal aorta for the radioimmunoassay of plasma secretin concentration. In the MA group, the exocrine pancreatic secretory response to 0.01 N HCI as well as saline infusion was significantly inhibited compared with that in the OC group. The pancreatic protein output of the MA group significantly decreased after the saline infusion and tended to decrease after the 0.01 N HCI infusion, compared with that of the OC group. However, there was no significant difference in plasma secretin concentration between the two groups. Therefore it is strongly suggested that the rat medial amygdaloid nucleus has a facilitatory influence on both basal and acid-stimulated pancreatic exocrine secretion, but the releasing mechanism of secretin appears not to be involved in the influence.
Exposure of skin cells, particularly keratinocytes to various nuclear factor-kappaB ($\textrm{NF}_{-{\kappa}}\textrm{B}$) activators [e.g. tumor necrosis factor-$\alpha$, interleukin-1, lipopolysaccharides, and ultraviolet light] leads to phosphorylation and degradation of the inhibitory protein, $\textrm{I}_{{\kappa}}\textrm{B}$. Liberated $\textrm{NF}_{-{\kappa}}\textrm{B}$ is translocated into the nucleus where it can change or alter expression of target genes, resulting in the secretion of extracellular signaling molecules including melanotrophic factors affecting melanocyte. In order to demonstrate the possible role of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation on the synthesis of melanotrophic factors from the keratinocytes, the activities of $\textrm{NF}_{-{\kappa}}\textrm{B}$ induced by melanogenic inhibitors (MIs) were determined in human HaCaT keratinocytes transfected with $\textrm{pNF}_{-{\kappa}}\textrm{B}$-SEAP-NPT plasmid. Transfectant cells released the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the $\textrm{NF}_{-{\kappa}}\textrm{B}$ activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selection marker for geneticin resistance. MIs such as niacinamide, kojic acid, hydroquinone, resorcinol, arbutin, and glycolic acid were preincubated with transfectant HaCaT cells for 3 h and then ultraviolet B (UVB) was irradiated. $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation was measured with the SEAP reporter gene assay using a fluorescence detection method. Of the Mis tested, kojic acid ($IC_{50}$/ = 60 $\mu$M) was found to be the most potent inhibitor of UVB-upregulating $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in transfectant HaCaT cells, which is followed by niacinamide ($IC_{50}$/= 540 $\mu$M). Pretreatment of the transfectant HaCaT cells with the Mis, especially kojic acid and niacinamide, effectively lowered $\textrm{NF}_{-{\kappa}}\textrm{B}$ binding measured by electrophoretic mobility shift assay. Furthermore, these two inhibitors remarkably reduced the secretion level of IL-6, one of melanotrophic factors, triggered by UV-radiation of the HaCaT cells. These observations suggest that Mis working at the in vivo level might act partially through the modulation of the synthesis of melanotrophic factors in keratinocyte.
Tuberculosis (TB) remains an enormous global health problem, and a new vaccine against TB more potent than the current inadequate BCG vaccine is urgently needed. We constructed three recombinant Mycobacterium bovis BCG (rBCG) strains over-expressing antigen (Ag) 85A, Ag85B, or both of M. tuberculosis using their own promoter and secretory sequence, or hsp60 promoter. SDS-PAGE analysis of rBCG proteins showed overexpression of Ag85A and Ag85B proteins in higher level than of those in their parental strain of BCG. In addition, rBCG(rBCG/B.FA) over-expressing Ag85A and Ag85B induced strong IFN-${\gamma}$ production in splenocytes. However, there was no significant difference in protective efficacy between rBCG and their parental BCG strain. In this study, therefore, rBCG over-expressing Ag85A, Ag85B, or both failed to show enhanced protection against M. tuberculosis infection in a mouse model.
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