Abstract We report a rare case of disseminated carcinomatosis of the bone marrow from rectal cancer with disseminated intravascular coagulation (DIC). A 65-year-old man was admitted with melena and low back pain at rest. X-ray examination showed rectal cancer with multiple bone metastases. Laboratory examination showed severe anemia and DIC. Histologic examination showed disseminated carcinomatosis of the bone marrow. The DIC was considered to be caused by disseminated carcinomatosis of the bone marrow from rectal cancer, and we immediately started treatment with anti-DIC therapy and anticancer chemotherapy with the modified FOLFOX6 regimen (mFOLFOX6). After some response to therapy, the patient's general condition deteriorated, and he died 128 days after admission. This is the first English report showing disseminated carcinomatosis of the bone marrow from colorectal cancer treated with mFOLFOX6.
Hypertension is a risk factor for renal impairment. While treatment of hypertension provides significant renal protection, there is still an unmet need requiring further exploration of additional pathogenetic mechanisms. We have found that the hypertension-related, calcium-regulated gene (HCaRG/COMMD5) is involved in renal repair. HCaRG is a small intracellular protein of 225 amino acids and its gene expression is negatively regulated by extracellular calcium concentrations. HCaRG is mostly expressed in the kidneys, with higher levels found in the spontaneously hypertensive rat than in normotensive rats. In an acute kidney injury model, HCaRG expression decreases immediately after injury but increases above baseline during the repair phase. In cell cultures, HCaRG has been shown to facilitate differentiation and to inhibit cell proliferation via p21 transactivation through the p53-independent signaling pathway. Induction of p21 independently of p53 is also observed in transgenic mice overexpressing HCaRG during the repair phase after ischemia/reperfusion injury, resulting in their improved renal function and survival with rapid re-differentiation of proximal tubular epithelial cells. In addition, transgenic mice recover rapidly from the inflammatory burst most likely as a result of maintenance of the tubular epithelial barrier. Recent studies indicate that facilitating re-differentiation and cell cycle regulation in injured renal proximal tubules might be important functions of HCaRG. We have proposed that HCaRG is a component of differential genetic susceptibility to renal impairment in response to hypertension.
Takasago has been devoted to producing l-menthol since 1954, and our long history of manufacturing this important aroma chemical is reviewed here. The current asymmetric catalytic process had its 30th anniversary in 2013. Our l-menthol process is considered carbon-neutral, and, therefore, 'green' and sustainable. It uses renewable myrcene obtained from gum rosin as a starting material. In addition, the Rh-BINAP (=2,2'-bis(diphenylphosphino)-1,1'-binaphthyl) catalytic system is highly efficient. This pathway not only leads l-menthol, but a variety of 100% biobased aroma chemical products as well. By measuring the (14) C levels in a material, one can determine the percentage of carbon that is biobased. This biobased assay, described as the ratio plant-derived C/fossil-derived C, can clarify how renewable a product really is. This will be highlighted for several of Takasago's key aroma chemicals.
Although many people have expressed alarm that we are witnessing a mass extinction, few projections have been quantified, owing to limited availability of time-series data on threatened organisms, especially plants. To quantify the risk of extinction, we need to monitor changes in population size over time for as many species as possible. Here, we present the world's first quantitative projection of plant species loss at a national level, with stochastic simulations based on the results of population censuses of 1618 threatened plant taxa in 3574 map cells of ca. 100 km2. More than 500 lay botanists helped monitor those taxa in 1994-1995 and in 2003-2004. We projected that between 370 and 561 vascular plant taxa will go extinct in Japan during the next century if past trends of population decline continue. This extinction rate is approximately two to three times the global rate. Using time-series data, we show that existing national protected areas (PAs) covering ca. 7% of Japan will not adequately prevent population declines: even core PAs can protect at best <60% of local populations from decline. Thus, the Aichi Biodiversity Target to expand PAs to 17% of land (and inland water) areas, as committed to by many national governments, is not enough: only 29.2% of currently threatened species will become non-threatened under the assumption that probability of protection success by PAs is 0.5, which our assessment shows is realistic. In countries where volunteers can be organized to monitor threatened taxa, censuses using our method should be able to quantify how fast we are losing species and to assess how effective current conservation measures such as PAs are in preventing species extinction.
Gene amplification and/or overexpression of the transcription factor c-MYC, which binds to the E-box sequence (5'-CACGTG-3'), has been observed in many human tumors. In this study, we have designed 5 pyrrole-imidazole (PI) polyamides recognizing E-box, and found that, among them, Myc-6 significantly suppresses malignant phenotypes of human osteosarcoma MG63 cells both in vitro and in vivo. Intriguingly, knockdown of the putative Myc-6 target MALAT1 encoding long noncoding RNA remarkably impaired cell growth of MG63 cells. Collectively, our present findings strongly suggest that Myc-6 exerts its tumor-suppressive ability at least in part through the specific down-regulation of MALAT1.
Adenosquamous carcinoma of the stomach is very rare; at present, there are only seven published reports. We report here an eighth case involving a 77-year-old Japanese man who was diagnosed with gastric cancer by upper endoscopy and computed tomography (CT). He underwent laparoscopic-assisted distal gastrectomy for early gastric cancer and the resected specimen was diagnosed as adenosquamous carcinoma limited to the submucosal layer. Only one lymph node metastasis was noted. Seven months later, liver metastasis (3 tumors, 15 mm maximum in diameter) was detected by abdominal CT. He was started on chemotherapy with S-1 and cisplatin (CDDP) and is alive 14 months after surgery. Almost all cases of adenosquamous carcinoma of the stomach are diagnosed in advanced stages and carry a very poor prognosis. Most patients with early adenosquamous carcinoma of the stomach survive for 2 or more years without recurrence, however our patient experienced recurrence 7 months after surgery. Therefore, future treatment for recurrent adenosquamous carcinoma of the stomach should be considered.
A questionnaire survey on postoperative adjuvant chemotherapy for gastric cancer was conducted for 76 hospitals affiliated with the Hiroshima Oncology Group of Gastric Cancer in Hiroshima prefecture in January 2011. Responses were obtained from 29 hospitals, including 12 core cancer treatment hospitals, and the following results were obtained. The percentage of patients completing 1 year of oral S-1 was >70%, affecting approximately 75% of the entire hospital cohort. Dose reduction was conducted in approximately 30% of patients because of age, poor PS, and renal insufficiency. The standard S-1 regimen (4 weeks of S-1 treatment followed by 2 weeks of rest)was adopted in almost half of the patients, whereas the rest of the patients received another treatment schedule such as 2 weeks of treatment followed by 1 week of rest. Dose reduction and withdrawal of S-1 due to adverse events were conducted more frequently in hospitals with low completion rates of 1-year S- 1 treatment than those with a high completion rate. S-1 was most commonly discontinued because of subjective adverse events and patient request, although the discontinuation rate according to objective adverse events such as bone marrow depression was not very high. The fact that some hospitals had high completion rates suggested the importance of supplementary tools for patient IC.
We have demonstrated that mesenchymal cells from spontaneously hypertensive rats genetically express complement 3 (C3). Mature tubular epithelial cells can undergo epithelial-to-mesenchymal transition (EMT) that is linked to the pathogenesis of renal fibrosis and injury. In this study, we investigated the contribution of C3 in EMT and in the renal renin-angiotensin (RA) systems associated with hypertension. C3a induced EMT in mouse TCMK-1 epithelial cells, which displayed increased expression of renin and Krüppel-like factor 5 (KLF5) and nuclear localization of liver X receptor ? (LXR?). C3 and renin were strongly stained in the degenerated nephrotubulus and colocalized with LXR? and prorenin receptor in unilateral ureteral obstruction (UUO) kidneys from wild-type mice. In C3-deficient mice, hydronephrus and EMT were suppressed, with no expression of renin and C3. After UUO, systolic blood pressure was increased in wild-type but not C3-deficient mice. In wild-type mice, intrarenal angiotensin II (ANG II) levels were markedly higher in UUO kidneys than normal kidneys and decreased with aliskiren. There were no increases in intrarenal ANG II levels after UUO in C3-deficient mice. Thus C3 induces EMT and dedifferentiation of epithelial cells, which produce renin through induction of LXR?. These data indicate for the first time that C3 may be a primary factor to activate the renal RA systems to induce hypertension.
Pyrrole-imidazole (PI) polyamides are a novel group of gene-silencing compounds, which bind to a minor groove of double stranded (ds)DNA in a sequence-specific manner. To explore the RNA binding properties of PI polyamides targeting rat transforming growth factor-?1 (TGF-?1 Polyamide) and influenza A virus (PA polyamide), we designed dsRNAs with an identical sequence to the target DNA and analyzed RNA binding properties of the polyamide. Biacore assay showed fast binding of TGF-?1 Polyamide to the dsRNA, whereas mismatch polyamide did not bind to the dsRNA. Dissociation equilibrium constant (KD) value was 6.7×10(-7) of the target dsRNA. These results indicate that PI polyamide could bind to RNA with a 2 log lower binding affinity than its DNA-binding affinity. We designed a PI polyamide targeting the panhandle stem region of influenza A virus. KD value of the PI polyamide to dsRNA targeting influenza A virus was 4.6×10(-7). Gel-shift assay showed that TGF-?1 and PA polyamides bound to the appropriate dsDNA, whereas these PI polyamides did not show obvious gel-shift with the appropriate dsRNA. Structural modeling suggests that PI polyamide binds to the appropriate B-form dsDNA in the minor groove, whereas it does not fit in the minor groove to dsRNA. Thus PI polyamides have a lower binding affinity with target dsRNA than they do with dsDNA. The distinct binding properties of PI polyamides to dsRNA and dsDNA may be associated with differences of secondary structure and chemical binding properties between target RNA and DNA.
Tooth tissue engineering offers very attractive perspectives for elaboration of regenerative treatments, which enables to cure tooth loss and restore quality of life of the patients. To elaborate such treatment, isolation and culture of dental pulp cell must be achieved as a key element. In this article, we report the establishment of a stable cell line from GFP transgenic rat dental pulp, named TGC (Tooth Matrix-forming, GFP Rat-derived Cell). TGCs have exhibited odontoblastic feature both in vitro and in vivo. In vitro, TGC exposed to osteogenic medium demonstrated collagen fiber synthesis with matrix vesicle and mineralization and formed a sheet-like substrate on the cell culture dish. Increased ALP activity and elevated transcription level of various genes involved in calcification and dentin formation were also observed. In vivo, transplanted TGC in SCID mice with ?-TCP particles formed dentin-like and pulp-like structure with lining odontoblast. Notably, even after up to 80 passages, TGCs retain their morphological features and differentiation ability. To our knowledge, this is the first report of a dental pulp-derived cell with such stable odontoblastic characteristics. TGC could be a very useful model for further study on dental pulp cell.
We investigate the effects of species interactions on the robustness of feedback control of the harvesting of prey species. We consider the consequences of feedback control of fishing effort. If a prey species is exploited, increasing fishing effort decreases predator abundance more than it does the prey abundance. Feedback control of fishing effort may cause the extinction of the predator, even if the prey population is well controlled. Even when fishing effort is controlled by predator density, it is difficult for the fishery and the predator to coexist, and, if they do so, the system exhibits complex dynamic behaviors. If the predator and fishery coexist, feedback control of fishing effort converges to a stable equilibrium, a synchronous cycle, or an asynchronous cycle. In the last case, the system undergoes more complex cycling with a longer period than that when the fishing effort is kept constant. These analyses suggest that there is no effective strategy that is robust against measurement errors, process errors and complex interactions in ecosystem dynamics.
The Pacific salmonid species Oncorhynchus mykiss is separated into a migratory form (steelhead trout) and a non-migratory form (rainbow trout). A decrease in water temperature is likely a cue triggering downstream behavior in the migratory form, and testosterone inhibits onset of this behavior. To elucidate differences in sensitivity to water temperature decreases between the migratory and non-migratory forms and effect of testosterone on the sensitivity, we examined two experiments. In experiment 1, we compared changes in body temperature during a short-term decrease in water temperature between both live and dead steelhead and rainbow trout. In experiment 2, we investigated effects of testosterone on body temperature decrease in steelhead trout. Water temperature was decreased by 3°C in 30min. The body temperature of the steelhead decreased faster than that of the rainbow trout. In contrast, there was no significant difference in the decrease in body temperature between dead steelhead and rainbow trout specimens. The body temperature of the testosterone-treated steelhead trout decreased more slowly than that of control fish. Our results suggest that the migratory form is more sensitive to decreases in water temperature than the non-migratory form. Moreover, testosterone might play an inhibitory role in sensitivity to such decreases.
Recent studies have established that, in benign tumors, a large number of cancer stem cells are present, which have great implications in tumor development. However, in ameloblastoma, a highly aggressive, locally invasive tumor with a high recurrence rate, whether or not cancer stem-like cells are present remains undetermined. Therefore, in this study we analyzed the protein expression of three candidate stem cell markers in ameloblastoma. Immunohistochemical staining for cancer stem cell (CSC) markers (CD133, CD44 and ABCG2) and for the proliferation marker Ki-67 was performed using 23 ameloblastoma cases. In all 23 samples, CD133, CD44 and ABCG2 were expressed. Nine (39.13%) cases showed high expression and 14 cases (60.87%) showed low expression for CD133. Twelve of the 23 cases (52.17%) showed high expression and 11 cases (47.83%) showed low expression for both CD44 and ABCG2, respectively. Ki-67 was mainly expressed in peripheral ameloblast-like cells, suggesting that these cells have a higher degree of differentiation and, therefore, are less likely to contain cancer stem-like cells. On the other hand, cells positive for CSC markers situated at the close proximity to peripheral cells were devoid of Ki-67 and may have the potential to be cancer stem-like cells. After analyzing the correlation between expression of three CSC markers with clinicopathological factors and Ki-67 expression, only CD44 expression was correlated with tumor recurrence (P=0.0391). In conclusion, this study showed various expression patterns of different types of cancer stem cell markers and the presence of candidate CSC-like cells in ameloblastoma, which are possibly involved in cell proliferation, tumor progression and recurrence.
Pyrrole-imidazole (PI) polyamide can bind to specific sequences in the minor groove of double-helical DNA and inhibit transcription of the genes. We designed and synthesized a PI polyamide to target the human connective tissue growth factor (hCTGF) promoter region adjacent to the Smads binding site. Among coupling activators that yield PI polyamides, 1-[bis(dimethylamino)methylene]-5-chloro-1H-benzotriazolium 3-oxide hexafluorophosphate (HCTU) was most effective in total yields of PI polyamides. A gel shift assay showed that a PI polyamide designed specifically for hCTGF (PI polyamide to hCTGF) bound the appropriate double-stranded oligonucleotide. A fluorescein isothiocyanate (FITC)-conjugated PI polyamide to CTGF permeated cell membranes and accumulated in the nuclei of cultured human mesangial cells (HMCs) and remained there for 48 h. The PI polyamide to hCTGF significantly decreased phorbol 12-myristate acetate (PMA)- or transforming growth factor-?1 (TGF-?1)-stimulated luciferase activity of the hCTGF promoter in cultured HMCs. The PI polyamide to hCTGF significantly decreased PMA- or TGF-?1-stimulated expression of hCTGF mRNA in a dose-dependent manner. The PI polyamide to hCTGF significantly decreased PMA- or TGF-?1-stimulated levels of hCTGF protein in HMCs. These results indicate that the developed synthetic PI polyamide to hCTGF could be a novel gene silencer for fibrotic diseases.
The repair of the kidney after ischemia/reperfusion injury involves proliferation of proximal tubular epithelial cells as well as cell migration and differentiation. Immediately after reperfusion, expression of hypertension-related calcium-regulated gene (HCaRG/COMMD5) decreases, but its expression increases even higher than baseline during repair. HCaRG inhibits proliferation and accelerates wound healing and differentiation in cultured cells, but whether HCaRG can stimulate renal repair after ischemia/reperfusion injury is unknown. Here, transgenic mice overexpressing human HCaRG survived longer and recovered renal function faster than littermate controls after ischemia/reperfusion (64% versus 25% survival at 7 days). Proliferation of proximal tubular epithelial cells stopped earlier after ischemia/reperfusion injury, E-cadherin levels recovered more rapidly, and vimentin induction abated faster in transgenic mice. HCaRG overexpression also reduced macrophage infiltration and inflammation after injury. Taken together, these data suggest that HCaRG accelerates repair of renal proximal tubules by modulating cell proliferation of resident tubular epithelial cells and by facilitating redifferentiation.
We formulated a full lifecycle bioenergetic model for bluefin tuna relying on the principles of Dynamic Energy Budget theory. Traditional bioenergetic models in fish research deduce energy input and utilization from observed growth and reproduction. In contrast, our model predicts growth and reproduction from food availability and temperature in the environment. We calibrated the model to emulate physiological characteristics of Pacific bluefin tuna (Thunnus orientalis, hereafter PBT), a species which has received considerable scientific attention due to its high economic value. Computer simulations suggest that (i) the main cause of different growth rates between cultivated and wild PBT is the difference in average body temperature of approximately 6.5°C, (ii) a well-fed PBT individual can spawn an average number of 9 batches per spawning season, (iii) food abundance experienced by wild PBT is rather constant and sufficiently high to provide energy for yearly reproductive cycle, (iv) energy in reserve is exceptionally small, causing the weight-length relationship of cultivated and wild PBT to be practically indistinguishable and suggesting that these fish are poorly equipped to deal with starvation, (v) accelerated growth rate of PBT larvae is connected to morphological changes prior to metamorphosis, while (vi) deceleration of growth rate in the early juvenile stage is related to efficiency of internal heat production. Based on these results, we discuss a number of physiological and ecological traits of PBT, including the reasons for high Feed Conversion Ratio recorded in bluefin tuna aquaculture.
Synthetic pyrrole-imidazole (PI) polyamides bind to the minor groove of double-helical DNA with high affinity and specificity, and inhibit the transcription of corresponding genes. We examined the effects of a transforming growth factor (TGF)-?1-targeted PI polyamide (Polyamide) on hypertrophic skin scars in rats. Hypertrophic scars were created dorsally in rats by incisions. FITC-labeled Polyamide was injected to investigate its distribution in the skin. Expression of TGF-?1, connective tissue growth factor (CTGF), collagen type1, and fibronectin mRNAs was evaluated by reverse transcription PCR analysis. The extent of fibrosis and the expression of TGF-?1 were evaluated histologically and immunohistochemically. Polyamide was distributed in almost all nuclei of skin cells. Expression of TGF-?1 mRNA reached a peak at 3 days after skin incision. Expression of CTGF and extracellular matrix mRNAs was increased continuously even after the peak induction of TGF-?1 mRNA. Injection of Polyamide completely inhibited both the development of scars and the induction of growth factors and extracellular matrix mRNAs. The treatment also markedly inhibited fibrotic changes and reduced the numbers of vimentin-positive spindle-shaped fibroblasts. Injection of Polyamide also reduced established hypertrophic scars in rats. Thus, TGF-?1-targeted PI polyamide should be a feasible gene silencer for hypertrophic scars and keloids.
The management programs for invasive species have been proposed and implemented in many regions of the world. However, practitioners and scientists have not reached a consensus on how to control them yet. One reason is the presence of various uncertainties associated with the management. To give some guidance on this issue, we characterize the optimal strategy by developing a dynamic model of invasive species management under uncertainties. In particular, focusing on (i) growth uncertainty and (ii) measurement uncertainty, we identify how these uncertainties affect optimal strategies and value functions. Our results suggest that a rise in growth uncertainty causes the optimal strategy to involve more restrained removals and the corresponding value function to shift up. Furthermore, we also find that a rise in measurement uncertainty affects optimal policies in a highly complex manner, but their corresponding value functions generally shift down as measurement uncertainty rises. Overall, a rise in growth uncertainty can be beneficial, while a rise in measurement uncertainty brings about an adverse effect, which implies the potential gain of precisely identifying the current stock size of invasive species.
Induced pluripotent stem cells (iPSCs) are novel stem cells derived from adult mouse and human tissues by reprogramming. Elucidation of mechanisms and exploration of efficient methods for their differentiation to functional cardiomyocytes are essential for developing cardiac cell models and future regenerative therapies. We previously established a novel mouse embryonic stem cell (ESC) and iPSC differentiation system in which cardiovascular cells can be systematically induced from Flk1(+) common progenitor cells, and identified highly cardiogenic progenitors as Flk1(+)/CXCR4(+)/VE-cadherin(-) (FCV) cells. We have also reported that cyclosporin-A (CSA) drastically increases FCV progenitor and cardiomyocyte induction from mouse ESCs. Here, we combined these technologies and extended them to mouse and human iPSCs. Co-culture of purified mouse iPSC-derived Flk1(+) cells with OP9 stroma cells induced cardiomyocyte differentiation whilst addition of CSA to Flk1(+) cells dramatically increased both cardiomyocyte and FCV progenitor cell differentiation. Spontaneously beating colonies were obtained from human iPSCs by co-culture with END-2 visceral endoderm-like cells. Appearance of beating colonies from human iPSCs was increased approximately 4.3 times by addition of CSA at mesoderm stage. CSA-expanded human iPSC-derived cardiomyocytes showed various cardiac marker expressions, synchronized calcium transients, cardiomyocyte-like action potentials, pharmacological reactions, and ultra-structural features as cardiomyocytes. These results provide a technological basis to obtain functional cardiomyocytes from iPSCs.
We conducted field surveys at 25 sites in three Japanese catchments to provide conservative estimates of the safe concentration of zinc (Zn) for the protection of riverine macroinvertebrate diversity. The relationships between the Zn concentration and six macroinvertebrate metrics for taxon richness were determined by using regression analysis; this included a piecewise regression model, where two lines are joined at an unknown point. For each metric the piecewise regression model with a zero slope below a threshold concentration was selected as the best model to explain the influence of Zn. Under the assumption that macroinvertebrate diversity reductions of <10% are acceptable, the safe concentrations of Zn were estimated to be 84, 115, 84, 80, 85, and 70 µg/L for total taxon richness, Ephemeroptera, Plecoptera, and Trichoptera (EPT) richness, mayfly richness, caddisfly richness, chironomid richness, and estimated total taxon richness at the riffle scale, respectively. These concentrations are more than twice the water quality standard for Zn in Japan (30 µg/L), suggesting that the standard is likely overprotective for macroinvertebrate diversity. Field studies are useful for evaluating the level of protectiveness of safe concentrations (water quality standards) based on individual-level effects from laboratory toxicity tests, and this evaluation process will have a crucial role in implementing more purpose-driven ecological risk managements that aim to protect natural populations and communities.
To establish whether the rates of surgical site infection (SSI) in gastrointestinal surgery are affected by the type of intra-abdominal suturing: sutureless, absorbable material (polyglactin: Vicryl), and silk.
Pyrrole-imidazole (PI) polyamides are small synthetic molecules that recognize and attach to the minor groove of DNA, thereby inhibiting gene transcription by blocking transcription factor binding. These derivatives can act as gene silencers inhibiting target gene expression under stimulatory conditions such as disease. To evaluate PI polyamides as treatments for the progression of renal diseases, we examined morphological effects, pharmacological properties, and the specificity of PI polyamides targeted to the transforming growth factor (TGF)-?1 promoter during salt-induced hypertensive nephrosclerosis in Dahl salt-sensitive rats. The targeted PI polyamide markedly reduced glomerulosclerosis and interstitial fibrosis without side effects. PI polyamide significantly decreased expression of TGF-?1 and extracellular matrix in the renal cortex. Microarray analysis found that only 3% of the transcripts were affected by PI polyamide, but this included decreased expression of extracellular matrix, TGF-?1-related cytokines, angiogenic, and cell stabilizing factors, proteinases, and renal injury-related factors. Thus, targeted PI polyamides are potential gene silencers for diseases not treatable by current remedies.
In this study, we develop a bioeconomic model of human alveolar echinococcosis (HAE) and formulate the optimal strategies for managing the infection risks in humans by applying optimal control theory. The model has the following novel features: (i) the complex transmission cycle of HAE has been tractably incorporated into the framework of optimal control problems and (ii) the volume of vermifuge spreading to manage the risk is considered a control variable. With this model, we first obtain the stability conditions for the transmission dynamics under the condition of constant control. Second, we explicitly introduce a control variable of vermifuge spreading into the analysis by considering the associated control costs. In this optimal control problem, we have successfully derived a set of conditions for a bang-bang control and singular control, which are mainly characterized by the prevalence of infection in voles and foxes and the remaining time of control. The analytical results are demonstrated by numerical analysis and we discuss the effects of the parameter values on the optimal strategy and the transmission cycle. We find that when the prevalence of infection in foxes is low and the prevalence of infection in voles is sufficiently high, the optimal strategy is to expend no effort in vermifuge spreading.
Invariant NKT cells (iNKT cells) play a pivotal role in the development of allergen-induced airway hyperresponsiveness (AHR) and inflammation. However, it is unclear what role they play in the initiation (sensitization) phase as opposed to the effector (challenge) phase. The role of iNKT cells during sensitization was examined by determining the response of mice to intratracheal transfer of OVA-pulsed or OVA-alpha-galactosylceramide (OVA/alphaGalCer)-pulsed bone marrow-derived dendritic cells (BMDCs) prior to allergen challenge. Wild-type (WT) recipients of OVA-BMDCs developed AHR, increased airway eosinophilia, and increased levels of Th2 cytokines in bronchoalveolar lavage fluid, whereas recipients of OVA/alphaGalCer BMDCs failed to do so. In contrast, transfer of these same OVA/alphaGalCer BMDCs into IFN-gamma-deficient (IFN-gamma(-/-)) mice enhanced the development of these lung allergic responses, which was reversed by exogenous IFN-gamma treatment following OVA-BMDC transfer. Further, Jalpha18-deficient recipients, which lack iNKT cells, developed the full spectrum of lung allergic responses following reconstitution with highly purified WT liver or spleen iNKT cells and transfer of OVA-BMDCs, whereas reconstituted recipients of OVA/alphaGalCer BMDCs failed to do so. Transfer of iNKT cells from IFN-gamma(-/-) mice restored the development of these responses in Jalpha18-deficient recipients following OVA-BMDC transfer; the responses were enhanced following OVA/alphaGalCer BMDC transfer. iNKT cells from these IFN-gamma(-/-) mice produced higher levels of IL-13 in vitro compared with WT iNKT cells. These data identify IFN-gamma as playing a critical role in dictating the consequences of iNKT cell activation in the initiation phase of the development of AHR and airway inflammation.
Pneumatosis intestinalis (PI) is a rare condition of unclear etiology affecting the intestinal tract; possible complications include obstruction, intussusception, volvulus, hemorrhage, and intestinal perforation. Pneumatosis intestinalis can occur as a primary idiopathic disease or as a condition secondary to various gastrointestinal and pulmonary diseases. This report presents an uncommon case of PI complicated with intussusception of the ascending colon in a male patient who underwent surgery for suspected lipomatosis that was histologically diagnosed as pneumatosis coli. Since the patient suffered from no other disorder, he was finally diagnosed as having primary PI. Physician awareness of this rare condition is essential in order to avoid a misdiagnosis and unnecessary surgical treatment.
To elucidate the roles of hyperpolarization-activated current (I(f)) in sinoatrial node (SAN) pacemaking, we theoretically investigated 1) the effects of I(f) on stability and bifurcation during hyperpolarization of SAN cells; 2) combined effects of I(f) and the sustained inward current (I(st)) or Na(+) channel current (I(Na)) on robustness of pacemaking against hyperpolarization; and 3) whether blocking I(f) abolishes pacemaker activity under certain conditions. Bifurcation analyses were performed for mathematical models of rabbit SAN cells; equilibrium points (EPs), periodic orbits, and their stability were determined as functions of parameters. Unstable steady-state potential region determined with applications of constant bias currents shrunk as I(f) density increased. In the central SAN cell, the critical acetylcholine concentration at which bifurcations, to yield a stable EP and quiescence, occur was increased by smaller I(f), but decreased by larger I(f). In contrast, the critical acetylcholine concentration and conductance of gap junctions between SAN and atrial cells at bifurcations progressively increased with enhancing I(f) in the peripheral SAN cell. These effects of I(f) were significantly attenuated by eliminating I(st) or I(Na), or by accelerating their inactivation. Under hyperpolarized conditions, blocking I(f) abolished SAN pacemaking via bifurcations. These results suggest that 1) I(f) itself cannot destabilize EPs; 2) I(f) improves SAN cell robustness against parasympathetic stimulation via preventing bifurcations in the presence of I(st) or I(Na); 3) I(f) dramatically enhances peripheral cell robustness against electrotonic loads of the atrium in combination with I(Na); and 4) pacemaker activity of hyperpolarized SAN cells could be abolished by blocking I(f).
We aimed to study kinetics of modulation by intracellular Mg(2+) of cardiac gap junction (Mg(2+) gate). Paired myocytes of guinea-pig ventricle were superfused with solutions containing various concentrations of Mg(2+). In order to rapidly apply Mg(2+) to one aspect of the gap junction, the non-junctional membrane of one of the pair was perforated at nearly the connecting site by pulses of nitrogen laser beam. The gap junction conductance (G(j)) was measured by clamping the membrane potential of the other cell using two-electrode voltage clamp method. The laser perforation immediately increased G(j), followed by slow G(j) change with time constant of 3.5 s at 10 mM Mg(2+). Mg(2+) more than 1.0 mM attenuated dose-dependently the gap junction conductance and lower Mg(2+) (0.6 mM) increased G(j) with a Hill coefficient of 3.4 and a half-maximum effective concentration of 0.6 mM. The time course of G(j) changes was fitted by single exponential function, and the relationship between the reciprocal of time constant and Mg(2+) concentration was almost linear. Based on the experimental data, a mathematical model of Mg(2+) gate with one open state and three closed states well reproduced experimental results. One-dimensional cable model of thirty ventricular myocytes connected to the Mg(2+) gate model suggested a pivotal role of the Mg(2+) gate of gap junction under pathological conditions.
Interstitial pneumonitis has sporadically been reported as a toxic effect of taxanes such as docetaxel and paclitaxel. This report describes 2 patients who developed interstitial pneumonitis after receiving chemotherapy including taxanes, and both cases grew serious enough to require respiratory support. The first case was a 57-year-old man with gastric cancer treated with docetaxel biweekly and S-1 for 2 weeks as adjuvant chemotherapy. After 4 courses of docetaxel, he presented acute dyspnea. The second case was a 66-year-old woman with breast cancer and postoperative pleural recurrence treated with weekly paclitaxel as fourth-line chemotherapy. She developed a dry cough, high fever, and dyspnea after 1 course of paclitaxel. In both cases, computed tomography (CT) showed extensive bilateral areas of ground-glass attenuation. They developed progressive interstitial infiltrates and respiratory failure that required mechanical ventilation. Taxane-induced interstitial pneumonitis was diagnosed to exclude other causes. From previous reports, intubation is associated with the survival of patients with taxane-induced interstitial pneumonitis. However, corticosteroid therapy was dramatically effective and resolved the interstitial pneumonitis in both our patients. Clinicians should be aware of this occasional complication during the course of chemotherapy with taxanes and initiate treatment, including respiratory support, as soon as possible.
This article reports development of a practical laboratory hard X-ray photoelectron spectroscopy (HXPS) system by combining a focused monochromatic Cr K(alpha) X-ray source, a wide angle acceptance objective lens and a high kinetic energy electron analyzer. The Cr K(alpha) source consists of a Cr target, a 15 kV focused electron gun, and a compact bent crystal monochromator. The X-ray spot size is variable from 10 microm (1.25 W) to 200 microm (50 W). A wide acceptance angle objective lens is installed in front of a hemispherical analyzer. The total resolution of 0.53 eV was obtained by Au Fermi-edge measurements. Angle acceptance of +/-35 degrees with angle resolution of 0.5 degrees was achieved by measuring Au 3d(5/2) peak through a hemicylinder multi-slit on an Au thin strip, in angle resolution mode. As the examples, silicon based multilayer thin films were used for showing the possibilities of deeper (larger) detection depth with the designed system.
Thrombosis is a major cause of morbidity and mortality, and thrombin is a major inducer of thrombus formation. Thus, several antithrombotic agents targeting thrombin have been developed. We previously reported on a thrombin derivative prepared by dual chemical modifications designated as M-anhydrothrombin, which possessed both anticoagulant and antiplatelet properties. In order to obtain a more potent antithrombotic thrombin derivative, we prepared a recombinant thrombin mutant and its chemically-modified derivative, and examined their antithrombotic efficacies.
Masting is synchronous intermittent production of seeds in perennial plant populations. Some self-compatible monoecious Quercus species, such as oaks, exhibit sex ratio dimorphism and produce a certain proportion of male flowers, even in a year when no seed set occurs. To investigate sex ratio dimorphism in masting trees, we introduced sexual allocation as an evolutionary trait into the Resource Budget Model and examined the evolution of the sex ratio. Analytical and numerical findings show that (1) perfectly synchronous intermittent reproduction does not evolve; (2) if the fruiting cost of female flowers R(c) is sufficiently large and the pollen limitation beta is intermediate, annual reproduction does not evolve; (3) under conditions (2), sex ratio dimorphism can evolve across a wide region of parameter space; (4) after dimorphism is established, individuals with a female-biased sex ratio receive much more pollen supply from male-biased individuals and tend to show intermittent reproduction with or without synchrony; and (5) dimorphism is maintained with irregular and nearly discontinuous changes of sex ratio. These results suggest that sex ratio dimorphism contributes to improving pollen availability and causes resource depletion and the occurrence of intermittent reproduction in female-biased individuals.
A number of studies have identified transforming growth factor-beta (TGF-beta) as a critical factor in renal diseases such as glomerulosclerosis and mesangioproliferative glomerulonephritis. TGF-beta stimulates proliferation of mesangial cells, production of extracellular matrix components and induces epithelial-mesenchymal transformation in renal tissue, which plays a critical role in the pathogenesis of renal injury. Thus, TGF-beta is a treatment target in renal diseases. However, progressive renal diseases cannot be cured with present medical technologies. We have developed ribozymes and a novel gene silencer pyrrole-imidazole polyamide targeted to TGF-beta that effectively ameliorate renal injury in hypertensive rats.
Corneal alkali burns are a serious clinical problem that often leads to permanent visual impairment. In this process, transforming growth factor (Tgf)-beta1 is upregulated and involved in the response to corneal injury and the process of corneal stromal scarring. To develop an efficient compound to inhibit Tgf-beta1 in the cornea, we designed GB1201, a pyrrole-imidazole (PI) polyamide targeting rat Tgf-beta1 gene promoter to the activator protein-1 (AP-1) binding site. GB1201 showed a high binding affinity to the target DNA sequence in the gel mobility shift and Biacore assays. GB1201 significantly inhibited the rat Tgf-beta1 gene promoter activity in HEK (human embryonic kidney) 293 cells in a concentration-dependent manner. Topically administrated GB1201 was distributed immediately to the nuclei of all cell layers of the cornea and remained for 24 hours. A corneal alkali burn model in rats was used to evaluate the therapeutic efficacy of GB1201. GB1201 suppressed the upregulation of Tgf-beta1 in the burned cornea, both in the mRNA and protein levels. Moreover, daily treatment with GB1201 for a week significantly improved the corneal tissue wound healing, reduced corneal stromal scarring, and prevented corneal haze formation. Our data suggest that PI polyamide may open new opportunities for therapeutic intervention in the treatment of chemically burned corneas.
Jagged1, a Notch ligand, and Notch have been implicated in Th2 differentiation, but their role in initiating IL-4 production and Th2 differentiation in vivo and the development of allergic airway responses has not been defined. In this study, we show that Jagged1 is up-regulated on bone marrow-derived dendritic cells (BMDCs) pulsed with allergen and that the transfer of these BMDCs before allergen challenge induces airway hyperresponsiveness (AHR) and eosinophilic airway inflammation. Treatment of CD4(+) T cells with a gamma-secretase inhibitor (GSI), which inhibits Notch signaling, resulted in decreased cytokine production when the cells were cocultured with allergen-pulsed, Jagged1-expressing BMDCs and, after the transfer of allergen-pulsed BMDCs, IL-4-deficient (IL-4(-/-)) recipients of GSI-treated naive CD4(+) T cells developed lower levels of AHR, reduced numbers of eosinophils, and lower Th2 cytokine levels when challenged with allergen. In vivo treatment of wild-type mice with Jagged1-Fc enhanced AHR and airway inflammation, whereas the transfer of BMDC transfected with Jagged1 small interfering RNA (siRNA) cells into WT or IL-4(-/-) mice before transfer of CD4(+) T cells resulted in decreased AHR, inflammation, and Th2 cytokines, indicating the critical role for Jagged1 expression on APCs. These data identify the essential role of the interactions between Notch on CD4(+) T cells and Jagged1 on APCs in the initiation of IL-4 production and Th2 differentiation for the development of AHR and allergic airway inflammation.
A 25-year-old man was referred to our hospital because of cough and an abnormal shadow on chest X-ray film showing bilateral hilar lymphadenopathy accompanied by multiple nodules in both lung fields. A transbronchial lung biopsy demonstrated non-caseating epithelioid cell granulomas, and we diagnosed sarcoidosis. He was observed without medication for 18 months, however, his chest X-ray film findings gradually worsened, and bilateral pleural effusion appeared. The pleural effusion consisted of exudative fluid with prominent lymphocytes, and ADA level was elevated to 57.0U/l. Thoracoscopy demonstrated multiple whitish granulations on the parietal and visceral pleura. The pleural biopsy specimens exhibited non-caseating epithelioid cell granulomas, and there was no evidence of acid-fast bacilli. Based on these findings, pleural sarcoidosis was diagnosed. He was treated with 30 mg oral prednisolone daily, and both pleural effusion and nodules of lung fields on chest X-ray film subsided. Sarcoidosis with bilateral pleural effusions is rare, and we discuss this condition in relation to the pertinent literature.
Naturally occurring Foxp3(+)CD4(+)CD25(+) T cells isolated from lungs of naive mice regulate lung allergic airway hyperresponsiveness, inflammation, levels of Th2 cytokines, and mucus production. OVA-specific (alphabetaTCR(+)) CD4(+)CD25(+) T cells suppressed ragweed-induced airway hyperresponsiveness and inflammation as did anti-TCR-treated OVA-specific CD4(+)CD25(+) T cells, suggesting that Ag-specificity was not required for expression of regulatory activities. Suppression was associated with increased levels of IL-10 and TGF-beta; decreased levels of IL-4, IL-5, and IL-13 in bronchoalveolar lavage fluid; and reduced recruitment and activation of CD8(+) T cells in the airways. Following intratracheal administration, OVA-specific CD4(+)CD25(+) T cells were identified in both the airway lumens and lung parenchyma, and in some instances in close proximity to host CD8(+) T cells. These results demonstrate that the regulatory activities of naturally occurring Foxp3(+)CD4(+)CD25(+) T cells on lung allergic responses are Ag-nonspecific and thus, independent of Ag-specific recognition.
Previous studies have shown that leukotriene B4 (LTB4), a proinflammatory lipid mediator, is linked to the development of airway hyperresponsiveness through the accumulation of IL-13-producing CD8+ T cells, which express a high affinity receptor for LTB4, BLT1 (Miyahara et al., Am J Respir Crit Care Med 2005;172:161-167; J Immunol 2005;174:4979-4984). By using leukotriene A4 hydrolase-deficient (LTA4H-/-) mice, which fail to synthesize LTB4, we determined the role of this lipid mediator in allergen-induced airway responses. Two approaches were used. In the first, LTA4H-/- mice and wild-type (LTA4H+/+) mice were systemically sensitized and challenged via the airways to ovalbumin. In the second, mice were passively sensitized with anti-ovalbumin IgE and exposed to ovalbumin via the airways. Mast cells were generated from bone marrow of LTA4H+/+ mice or LTA4H-/- mice. After active sensitization and challenge, LTA4H-/- mice showed significantly lower airway hyperresponsiveness compared with LTA4H+/+ mice, and eosinophil numbers and IL-13 levels in the bronchoalveoloar lavage of LTA4H-/- mice were also significantly lower. LTA4H-/- mice also showed decreased airway reactivity after passive sensitization and challenge. After LTA4H+/+ mast cell transfer, LTA4H-/- mice showed increased airway reactivity after passive sensitization and challenge, but not after systemic sensitization and challenge. These data confirm the important role for LTB4 in the development of altered airway responses and suggest that LTB4 secretion from mast cells is critical to eliciting increased airway reactivity after passive sensitization with allergen-specific IgE.
We examined the influence of heavy metal pollution from an abandoned mine on benthic macroinvertebrates, at population and community levels, and the potential amount of food available for drift-feeding fish in northern Japanese streams. We studied multiple polluted and unpolluted sites with similar longitudinal positions to avoid problems caused by upstream-downstream comparisons. The ranges of zinc, copper, cadmium, and lead concentrations among the study sites were 5 to 812 microg/L, less than 0.12 to 5.2 microg/L, less than 0.0026 to 4.9 microg/L, and 0.1 to 18.6 microg/L, respectively. The abundance of several populations and community metrics showed a significant negative response to heavy metal pollution. Mayfly diversity and abundance was relatively sensitive to heavy metal pollution. In addition, the biomass of groups of macroinvertebrate taxa that are highly available for salmonids were significantly reduced at metal-polluted sites; this decrease in the most highly available group was noticeable (99% at the heavily polluted upper sites and 69% at the moderately polluted lower sites in spring). These results suggest that we should consider the indirect effect of pollution on food availability for the conservation of fish populations that depend on drifting macroinvertebrates.
Hepatitis E virus (HEV) genotype 3, which usually causes asymptomatic infection in Japan, induced severe hepatitis in 8 patients. To better understand genetic features of HEV associated with increased virulence, we determined the complete or near-complete nucleotide sequences of HEV from these 8 patients and from 5 swine infected with genotype 3 strain swJ19. Phylogenetic analysis showed that the isolates from the 8 patients and the 5 swine grouped separately from the other genotype 3 isolates to create a unique cluster, designated JIO. The human JIO-related viruses encoded 18 amino acids different from those of the other HEV genotype 3 strains. One substitution common to almost all human HEV strains in the JIO cluster was located in the helicase domain (V239A) and may be associated with increased virulence. A zoonotic origin of JIO-related viruses is suspected because the isolates from the 5 swine also possessed the signature V239A substitution in helicase.
gammadelta T cells regulate airway reactivity, but their role in ozone (O3)-induced airway hyperresponsiveness (AHR) is not known. Our objective was to determine the role of gammadelta T cells in O3-induced AHR. Different strains of mice, including those that were genetically manipulated or antibody-depleted to render them deficient in total gammadelta T cells or specific subsets of gammadelta T cells, were exposed to 2.0 ppm of O3 for 3 hours. Airway reactivity to inhaled methacholine, airway inflammation, and epithelial cell damage were monitored. Exposure of C57BL/6 mice to O3 resulted in a transient increase in airway reactivity, neutrophilia, and increased numbers of epithelial cells in the lavage fluid. TCR-delta(-/-) mice did not develop AHR, although they exhibited an increase in neutrophils and epithelial cells in the lavage fluid. Similarly, depletion of gammadelta T cells in wild-type mice suppressed O3-induced AHR without influencing airway inflammation or epithelial damage. Depletion of Vgamma1+, but not of Vgamma4+ T cells, reduced O3-induced AHR, and transfer of total gammadelta T cells or Vgamma1+ T cells to TCR-delta(-/-) mice restored AHR. After transfer of Vgamma1+ cells to TCR-delta(-/-) mice, restoration of AHR after O3 exposure was blocked by anti-TNF-alpha. However, AHR could be restored in TCR-delta(-/-)mice by transfer of gammadelta T cells from TNF-alpha-deficient mice, indicating that another cell type was the source of TNF-alpha. These results demonstrate that TNF-alpha and activation of Vgamma1+ gammadelta T cells are required for the development of AHR after O3 exposure.
Pyrrole-imidazole polyamide can be combined in antiparallel side-by-side dimeric complexes along the minor groove of DNA in a sequence-specific manner. Pyrrole-imidazole polyamides are effective inhibitors of transcription factors as well as viral repressors and transactivators. Recently, lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) was reported to be a major factor contributing to the pathogenesis of coronary atherosclerosis. In this study, we designed a pyrrole-imidazole polyamide specific for the LOX-1 gene and evaluated its effect on LOX-1 gene transcription. A pyrrole-imidazole polyamide was designed to target the AP-1 binding site of the LOX-1 gene and synthesized by solid phase methods. This pyrrole-imidazole polyamide significantly inhibited LOX-1 promoter activity in HEK293 cells, determined by the luciferase assay. LOX-1 mRNA expression was also inhibited by the pyrrole-imidazole polyamide at a concentration of 10-9 mol/l in human umbilical vein endothelial cells (HUVEC), determined by the real-time PCR method. HUVEC were treated by pyrrole-imidazole polyamide targeting the LOX-1 gene, and apoptosis was assessed using Hoechst stain, terminal deoxy nucleotidyl transferase-mediated UTP end labeling method, and dye-uptake bioassay. Treatment of HUVEC for 72 h with LOX-1 targeted pyrrole-imidazole polyamide decreased apoptosis induced by angiotensin II and oxidized low-density lipoprotein (ox-LDL) loading in all assays. This novel therapeutic agent, pyrrole-imidazole polyamide, could specifically inhibit LOX-1 gene expression by reducing the promoter activity of the gene. Pyrrole-imidazole polyamide seems to be a powerful promising new agent that can be used to explore therapies based on inhibition of transcription. Molecular recognition of DNA by small molecules could provide insight into the development of new human medicines.
The problem of controlling invasive species has emerged as a global issue. In response to invasive species threats, governments often propose eradication. This article challenges the eradication view by studying optimal strategies for controlling invasive species in a simple dynamic model. The analysis mainly focuses on deriving policy implications of catchability in a situation where a series of controlling actions incurs operational costs that derive from the fact that catchability depends on the current stock size of invasive species. We analytically demonstrate that the optimal policy changes drastically, depending on the sensitivity of catchability in response to a change in the stock size, as well as on the initial stock. If the sensitivity of catchability is sufficiently high, the constant escapement policy with some interior target level is optimal. In contrast, if the sensitivity of catchability is sufficiently low, there could exist a threshold of the initial stock which differentiates the optimal action between immediate eradication and giving-up without any control. In the intermediate range, immediate eradication, giving-up without any control, or more complex policies may be optimal. Numerical analysis is employed to present economic intuitions and insights in both analytically tractable and intractable cases.
We report two cases of pneumonitis caused by Seishinrenshiin. A 54-year-old woman and a 80-year-old man had taken Seishinrenshiin for cystitis and benign prostatic hypertrophy. Their chest radiograph showed diffuse ground-glass shadows in the whole lung fields and chest CT showed diffuse ground-glass-opacities predominantly in the lower lung fields of both lungs. Biochemical tests revealed mild liver dysfunction and inflammatory reactions. Their abnormal chest shadows disappeared after discontinuation of Seishinrenshiin. We should be aware that Seishinrenshiin, as well as other Chinese herbal medicine, could be cause of drug-induced pneumonitis.
The diagnosis of Pneumocystis pneumonia (PCP) is based on microscopic examination of respiratory specimens. PCP patients without AIDS have a lower burden of P. jiroveci than those with AIDS, which leads to difficulty in detecting the organisms. Although conventional PCR (c-PCR) has been used to detect the DNA, it is frequently positive in patients with colonization. Real-time PCR (r-PCR), a method to detect the DNA quantitatively, might be helpful in distinguishing between infection and colonization. We investigated the utility of real-time PCR in the diagnosis of PCP in non-AIDS patients.
A 67-year-old woman was admitted to our hospital for weakness in her right hand. MRI showed multiple cerebral infarctions and ultrasonic cardiography revealed vegetation on her aortic valve, so embolic stroke was diagnosed. Though she was afebrile and her vital signs were normal, chest CT revealed several enlarged mediastinal lymph nodes and a small nodule in the left lower lobe of the lung. Stage III adenocarcinoma of the lung was diagnosed, and the cause of her cerebral infarctions was found to be nonbacterial thrombotic endocarditis (NBTE). NBTE is known as the cause of embolic stroke among patients with advanced cancer, particularly adenocarcinoma. Prompt initiation of continuous heparin administration is required to interrupt the progress of cerebral thromboembolism in NBTE. In cases of coexisting cancer and embolic stroke, we should consider the possibility of NBTE.
Though cardiac progenitor cells should be a suitable material for cardiac regeneration, efficient ways to induce cardiac progenitors from embryonic stem (ES) cells have not been established. Extending our systematic cardiovascular differentiation method of ES cells, here we show efficient and specific expansion of cardiomyocytes and highly cardiogenic progenitors from ES cells. An immunosuppressant, cyclosporin-A (CSA), showed a novel effect specifically acting on mesoderm cells to drastically increase cardiac progenitors as well as cardiomyocytes by 10-20 times. Approximately 200 cardiomyocytes could be induced from one mouse ES cell using this method. Expanded progenitors successfully integrated into scar tissue of infracted heart as cardiomyocytes after cell transplantation to rat myocardial infarction model. CSA elicited specific induction of cardiac lineage from mesoderm in a novel mesoderm-specific, NFAT independent fashion. This simple but efficient differentiation technology would be extended to induce pluripotent stem (iPS) cells and broadly contribute to cardiac regeneration.
Dendritic cells (DCs) are considered to be the most efficient antigen-presenting cells. Intratracheal administration of allergen-pulsed bone marrow-derived dendritic cells (BMDCs) before allergen challenge induces airway hyperresponsiveness (AHR) and inflammation. Ovalbumin (OVA)-pulsed BMDCs from wild-type (WT) mice were transferred into naive WT, CD4(-/-), CD8(-/-), or IL-13(-/-) mice. Two days (short protocol) or 10 days (long protocol) after BMDC transfer, mice were challenged with 1% OVA for 3 days and assayed 2 days later. Transfer of OVA-primed BMDCs into BALB/c or C57BL/6 mice elicited AHR in both protocols. Airway eosinophilia, Th2 cytokines, or goblet cell metaplasia were increased in the long but not short protocol. Lung T cells from both protocols produced Th2 cytokines in response to OVA in vitro. Carboxyfluorescein diacetate succinimidylester-labeled BMDCs were observed in bronchoalveolar lavage (BAL) fluid and lung parenchyma at early time points, and were detected in draining lymph nodes 48 hours after transfer. CD8(-/-) mice developed AHR comparable to WT mice in the short protocol, but decreased levels of AHR, airway eosinophilia, Th2 cytokines in BAL fluid, and goblet cell metaplasia compared with WT mice in the long protocol. CD4(-/-) or IL-13(-/-) mice did not develop AHR or airway inflammation in either protocol. These data suggest that allergen-pulsed BMDCs initiate development of AHR that is dependent initially on CD4(+) T cells, and at later time periods on CD8+ T cells and IL-13. Thus, the timing of allergen challenge after transfer of allergen-pulsed BMDC affects the development of AHR and airway inflammation.
Although the use of drug-eluting stents (DESs) has been shown to limit neointima hyperplasia, currently available DESs may adversely affect re-endothelialization. To evaluate whether a novel gene silencer pyrrole-imidazole (PI) polyamide targeting transforming growth factor (TGF)-beta1 is a candidate agent for the DESs, we examined the effects of PI polyamide targeting the TGF-beta1 promoter on neointimal formation in rat carotid artery after balloon injury.
The effect of bacterial cell-surface hydrophobicity on the bioconversion of water-immiscible chemicals in an aqueous-organic (A/O) two-liquid-phase culture system was investigated. Escherichia coli JM109 and Rhodococcus opacus B-4 were used as hydrophilic and hydrophobic whole-cell catalysts, respectively. Hydroxylation reactions of monoaromatics, including toluene (log P(ow)=2.9), ethylbenzene (3.1), n-propylbenzene (3.4), and sec-butylbenzene (3.7), were employed as model conversions. When the todC1C2BA genes encoding Pseudomonas putida toluene dioxygenase were expressed in E. coli JM109, the yield of hydroxylated monoaromatics decreased with increasing substrate hydrophobicity. By contrast, R. opacus transformants, which expressed the todC1C2BA genes, showed high performance in the hydroxylation of monoaromatics, irrespective of substrate hydrophobicity. When the R. opacus transformants were examined for their ability to hydroxylate monoaromatics in an aqueous single-liquid-phase culture, the reaction velocity was markedly lower than that observed in the A/O two-liquid-phase culture. These results suggested that R. opacus B-4 accessed the hydrophobic substrates in the oil phase, thus making it more effective for the bioconversion reactions.
Fc? receptors I and III are thought to be involved in the development of lupus nephritis. Expression of Fc receptor common gamma chain (FcR?) is necessary for the stable expression of Fc? receptors I and III. The aim of this study was to develop a novel agent for the treatment of immune complex related renal disease using a gene regulator, pyrrole(Py)-imidazole(Im) (PI) polyamide, targeting the mouse FcR? gene promoter. Two PI polyamides targeting FcR? promoters were designed and synthesized. The effect of the PI polyamides on FcR? mRNA expression was evaluated in J774.A cells by real-time polymerase chain reaction (PCR), and CD16/32 protein expression was determined by immunocytochemical analysis and flow cytometry. The effects of these polyamides on FcR? gene expression and CD16/32 protein expression were evaluated in mouse peripheral blood mononuclear cells (PBMCs). One milligram per kilogram body weight of PI polyamide was injected via the tail vein every 2 d for 1 week and PBMCs were collected and analyzed. PI polyamide showed a specific binding to the target DNA in a gel mobility shift assay. Treatment of J774.A cells with 1.0 µM PI polyamide 1 significantly reduced FcR? mRNA expression and CD16/32 surface protein expression in J774.A cells. Similarly, PI polyamide significantly decreased expression of FcR? mRNA and CD16/32 in the PBMCs of C57B6 mice. PI polyamide designed to bind the FcR? promoter decreased FcR? gene and CD16/32 protein expression. PI polyamide targeting the FcR? gene may be a novel gene regulator for the prevention of lupus nephritis or other immune complex-related disease.
Thymoma is an epithelial neoplasm of the thymus, which commonly lies in the anterior mediastinum and, therefore, an intrathoracic origin is considered to be rare. This report presents a case of giant thymoma arising in the thoracic cavity. A 61-year-old male presented with a chronic cough and breathlessness. Chest CT revealed a well enhanced giant mass approximately 18?cm in diameter in the right thoracic cavity. FDG-PET showed that the SUVmax of the tumor was 5.0 in the center and almost 2.5 in the surrounding area. A trans- bronchial needle biopsy was performed to find atypical cells. Surgery was scheduled based on the radiological and histological findings. A well-defined giant mass in the thoracic cavity, measuring 18?×?14.5?×?11?cm had undergone expansive growth without apparent invasion. The tumor was completely resected without combined resection of the other organs. The weight of the tumor was 1350?g. The tumor was histologically diagnosed to be type AB thymoma according to the World Health Organization classification and Masaoka stage IIB.
The purpose of this study was to determine the poor prognostic factors after repeat hepatectomy (Hx) in patients with recurrent hepatocellular carcinoma (HCC). Overall survival rates and clinicopathological variables in 112 patients with repeat Hx from 1992 to 2010 were compared with those in 531 patients who underwent a primary Hx. To clarify the poor prognosis factors after repeat Hx, survival data among 112 patients were univariately and multivariately analyzed. Overall survival after repeat Hx was similar for that of the patients who underwent a primary Hx. The mean age of repeat Hx group was significantly higher, and a well-preserved liver function was recognized than the primary Hx group. Multivariate analysis revealed that: 1) indocyanine green retention rate at 15 minutes; 2) disease-free interval; 3) tumor size; 4) portal vein invasion at primary Hx; 5) gender; and 6) estimated blood loss to be an independent and significant poor prognostic factors. The overall 3-year postrecurrence overall survival rates were 100, 91.3, 59.6, and 0 per cent at risk number (R) R0, R1/2, R3, R4, respectively (P < 0.05). Repeat Hx provided a good compatible prognosis with primary Hx. In our findings, five risk factors to predict poor outcomes after repeat Hx were useful. Patients with recurrent HCC do not have universally poor outcomes, and our simple scoring system using five poor prognostic factors could serve to advise the prognosis and the potential benefit for patient selection about repeat Hx.
The effectiveness of subcuticular absorbable suture with subcutaneous drainage to decrease the risk of postoperative incisional surgical site infection (SSI) in hepatocellular carcinoma (HCC) patients was evaluated.
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