Enhanced adsorption of arsenic using calcined alginate bead containing alum sludge from water treatment facilities.

An adsorbent of bead type to remove arsenic (As) was developed by calcination of sodium alginate and polyvinyl alcohol containing a powder form of alum sludge. The adsorbent was evaluated in terms of adsorption kinetics, capacities in batch tests and by a column study. The calcination process created rough surface and increased the surface area of bead 100 times, which enhanced the adsorption kinetics of As onto the calcined adsorbent 3-21 times than un-calcined bead. However, the adsorption capacity decreased slightly compared to the un-calcined adsorbent. The column study showed similar adsorption capacity with commercial adsorbent and powder form of alum sludge considering the standard value of As for drinking water. The calcination process enhanced the adsorption kinetics of the adsorbent for As removal, one of major barrier of bead type adsorbent compared to powder type, which could reduce the bed volume of the reactor.

Discovery of a hyperalkaline liquid condensed phase: significance toward applications in carbon dioxide sequestration.

Bicarbonate ion-containing solutions such as seawater, natural brines, bovine serum and other mineralizing fluids have been found to contain hyperalkaline droplets of a separate, liquid condensed phase (LCP), that have higher concentrations of bicarbonate ion (HCO3 -) relative to the bulk solution in which they reside. The existence and unique composition of the LCP droplets have been characterized by nanoparticle tracking analysis, nuclear magnetic resonance spectroscopy, fourier transform infrared spectroscopy, dissolved inorganic carbon analysis and refractive index measurements. Carbon dioxide can be brought into solution through an aqueous reaction to form LCP droplets that can then be separated by established industrial membrane processes as a means of concentrating HCO3 -. Reaction of calcium with the LCP droplets results in calcium carbonate precipitation and mineral formation. The LCP phenomenon may bear on native mineralization reactions and has the potential to change fundamental approaches to carbon capture, sequestration and utilization.

Diverse and Synergistic Actions of Phytochemicals in a Plant-Based Multivitamin/Mineral Supplement against Oxidative Stress and Inflammation in Healthy Individuals: A Systems Biology Approach Based on a Randomized Clinical Trial.

Traditional clinical methodologies often fall short of revealing the complex interplay of multiple components and targets within the human body. This study was designed to explore the complex and synergistic effects of phytochemicals in a plant-based multivitamin/mineral supplement (PBS) on oxidative stress and inflammation in healthy individuals. Utilizing a systems biology framework, we integrated clinical with multi-omics analyses, including UPLC-Q-TOF-MS for 33 phytochemicals, qPCR for 42 differential transcripts, and GC-TOF-MS for 17 differential metabolites. A Gene Ontology analysis facilitated the identification of 367 biological processes linked to oxidative stress and inflammation. As a result, a comprehensive network was constructed consisting of 255 nodes and 1579 edges, featuring 10 phytochemicals, 26 targets, and 218 biological processes. Quercetin was identified as having the broadest target spectrum, succeeded by ellagic acid, hesperidin, chlorogenic acid, and quercitrin. Moreover, several phytochemicals were associated with key genes such as HMOX1, TNF, NFE2L2, CXCL8, and IL6, which play roles in the Toll-like receptor, NF-kappa B, adipocytokine, and C-type lectin receptor signaling pathways. This clinical data-driven network system approach has significantly advanced our comprehension of a PBS's effects by pinpointing pivotal phytochemicals and delineating their synergistic actions, thus illuminating potential molecular mechanisms.

Blood Microbiota Profile Is Associated with the Responsiveness of Postprandial Lipemia to Platycodi radix Beverage: A Randomized Controlled Trial in Healthy Subjects.

Prolonged postprandial hyperlipidemia may cause the development of cardiovascular diseases. This study explored whether postprandial triglyceride-rich lipoprotein (TRL) clearance responsiveness to Platycodi radix beverage (PR) is associated with changes in blood microbiota profiles. We conducted an 8-week randomized controlled clinical trial involving normolipidemic adults with low fruit and vegetable intakes. Participants underwent an oral fat tolerance test and 16S amplicon sequencing analysis of blood microbiota. Using the Qualitative Interaction Trees, we identified responders as those with higher baseline dietary fat intake (>38.5 g/day) and lipoprotein lipase levels (>150.6 ng/mL), who showed significant reductions in AUC for triglyceride (TG) and chylomicron-TG after the oral fat tolerance test. The LEfSe analysis showed differentially abundant blood microbiota between responders and non-responders. A penalized logistic regression algorithm was employed to predict the responsiveness to intervention on the TRL clearance based on the background characteristics, including the blood microbiome. Our findings suggest that PR intake can modulate postprandial TRL clearance in adults consuming higher fat intake over 38.5 g/day and low fruit and vegetable intake through shared links to systemic microbial signatures.

Analgesic effects of dexmedetomidine in vincristine-evoked painful neuropathic rats.

Dexmedetomidine, which is a selective α2-adrenoceptor agonist, was recently introduced into clinical practice for its analgesic properties. The purpose of this study was to evaluate the effects of dexmedetomidine in a vincristine-evoked neuropathic rat models. Sprague-Dawley rats were injected intraperitoneally with vincristine or saline (0.1 mg/kg/day) using a 5-day-on, 2-day-off schedule for 2 weeks. Saline and dexmedetomidine (12.5, 25, 50, and 100 µg/kg) were injected to rats developed allodynia 14 days after vincristine injection, respectively. We evaluated allodynia at before, 15, 30, 60, 90, 120, 180, and 240 min, and 24 hr after intraperitoneal drug (normal saline or dexmedetomidine) injection. Saline treatment did not show any differences for all the allodynia. Maximal paw withdrawal thresholds to mechanical stimuli were 3.0 ± 0.4, 9.1 ± 1.9, 13.0 ± 3.6, 16.6 ± 2.4, and 24.4 ± 1.6 g at saline, 12.5, 25, 50, and 100 µg/kg dexmedetomidine injection, respectively. Minimal withdrawal frequency to cold stimuli were 73.3 ± 4.2, 57.1 ± 6.8, 34.3 ± 5.7, 20.0 ± 6.2, and 14.3 ± 9.5 g at saline, 12.5, 25, 50, and 100 µg/kg dexmedetomidine injection, respectively. Dexmedetomidine shows a dose-dependent antiallodynic effect on mechanical and cold stimuli in vincristine-evoked neuropathic rat models (P < 0.05).

Meta-Analysis of Randomized Clinical Trials Evaluating Effectiveness of a Multivitamin Supplementation against Oxidative Stress in Healthy Subjects.

A meta-analysis has been widely applied to draw general conclusions using a set of studies with similar purposes and designs. This study aimed to perform a meta-analysis of six randomized placebo-controlled trials, independently conducted for the relationship between a plant-based multivitamin/mineral supplementation (PMS) and oxidative stress for 6 to 8 weeks, to provide overall estimates of those effects. In detail, linear mixed model analysis was first conducted on each study to obtain individual estimates; then, two types of meta-analysis were applied to combine the individual estimates from all available studies (overall meta-analysis) and region-specific studies (subgroup meta-analysis). In the meta-analysis, we selected 19 biomarker variables that overlapped in at least two studies and found 6 variables significant in at least one meta-analysis. The overall estimates of beta coefficients were 0.17 for vitamin C, 0.80 for vitamin B6, 0.46 for vitamin B12, 0.81 for folate, 0.36 for ß-carotene, and -0.17 for oxidized LDL (ox-LDL). Subsequent association analysis revealed significant negative correlations between plasma free radical scavenging nutrients and plasma ox-LDL levels, indicating a general benefit of PMS in alleviating oxidative stress by providing exogenous oxidant scavengers.

Yellow Yeast Rice Prepared Using Aspergillus terreus DSMK01 Lowers Cholesterol Levels by Stimulating Bile Salt Export Pump in Subjects with Mild-to-Moderate Hypercholesterolemia: A Randomized Controlled Trial.

SCOPE: Aspergillus terreus is an industrial microorganism used in the brewing and sauce industries. It produces monacolin K, a natural statin. The study conducted an 8-week randomized controlled trial with hypercholesterolemic subjects to examine the hypocholesterolemic effects and mechanisms of supplementation with yellow yeast rice (YYR) prepared by growing Aspergillus fungi on steamed rice. METHODS AND RESULTS: YYR supplementation markedly reduced total cholesterol, LDL, and apolipoprotein B100 levels in plasma compared with the placebo. In addition, YYR induced a significantly increased ATP binding cassette subfamily B member 11 (ABCB11) gene expression compared with the placebo, indicating the role of YYR in lowering intrahepatic cholesterol availability by stimulating the bile salt export pump. Upregulation of LDL receptor (LDLR) and 3-methylglutaryl-CoA reductase (HMGCR) gene expressions provided additional evidence to support the role of YYR in reducing hepatic cholesterol availability. Plasma metabolomic profiling revealed the possibility of diminishing bile acid absorption. Finally, Spearman rank analysis showed correlations of plasma cholesterol profiles with HMGCR and LDLR gene expressions (negative) and plasma bile acids (positive). Plasma bile acids also correlated with ABCB11 (negative) and LDLR (positive) gene expressions. CONCLUSION: These findings suggest that daily YYR supplementation exerted hypocholesterolemic effects in mild-to-moderate hypercholesterolemic subjects by reducing intrahepatic cholesterol availability through stimulating bile salt export pumps and inhibiting cholesterol biosynthesis.

Blood-derived iron mediates free radical production and neuronal death in the hippocampal CA1 area following transient forebrain ischemia in rat.

Abnormal brain iron homeostasis has been proposed as a pathological event leading to oxidative stress and neuronal injury under pathological conditions. We examined the possibility that neuronal iron overload would mediate free radical production and delayed neuronal death (DND) in hippocampal CA1 area after transient forebrain ischemia (TFI). Mitochondrial free radicals (MFR) were biphasically generated in CA1 neurons 0.5-8 and 48-60 h after TFI. Treatment with Neu2000, a potent spin trapping molecule, as well as trolox, a vitamin E analogue, blocked the biphasic MFR production and attenuated DND in the CA1, regardless of whether it was administered immediately or even 24 h after reperfusion. The late increase in MFR was accompanied by iron accumulation and blocked by the administration of deferoxamine-an iron chelator. Iron accumulation was attributable to prolonged upregulation of the transferrin receptor and to increased uptake of peripheral iron through a leaky blood-brain barrier. Infiltration of iron-containing cells and iron accumulation were attenuated by depletion of circulating blood cells through X-ray irradiation of the whole body except the head. The present findings suggest that excessive iron transported from blood mediates slowly evolving oxidative stress and neuronal death in CA1 after TFI, and that targeting iron-mediated oxidative stress holds extended therapeutic time window against an ischemic event.

Exfoliated and reorganized graphite oxide on titania nanoparticles as an auxiliary co-catalyst for photocatalytic solar conversion.

The hybrid of graphite oxide (GO)/TiO(2) was prepared through the spontaneous exfoliation of bulky graphite oxide and reorganization with TiO(2) nanoparticles as a solar conversion and hydrogen-generating photocatalyst. GO/TiO(2) showed enhanced activities for both photocurrent generation (in an electrode form) and hydrogen production (in a slurry form) than those of bare TiO(2) under UV light irradiation. The enhanced photocatalytic activity of GO/TiO(2) is ascribed to the ability of graphitic layers in accepting and transporting electrons from excited TiO(2), promoting the charge separation. When GO was hybridized with platinized TiO(2) (Pt/TiO(2)), it showed a marked synergistic effect for the photocatalytic hydrogen production compared with GO/TiO(2) and Pt/TiO(2). This indicates that the cheap and abundant carbon material can be a good candidate for an electron attracting reservoir and an auxiliary co-catalyst for the photocatalytic hydrogen production.

Differential Proximity of Perisynaptic Astrocytic Best1 at the Excitatory and Inhibitory Tripartite Synapses in APP/PS1 and MAOB-KO Mice Revealed by Lattice Structured Illumination Microscopy.

Bestrophin-1 (Best1) is a GABA- and glutamate-permeable, Ca2+-activated Cl- channel, which is mainly expressed in astrocytes and localized at the microdomain or perisynaptic junction of the tripartite synapse. Distribution of Best1 is dramatically changed in pathological conditions such as Alzheimer's disease. However, it is still unknown whether Best1 is located at the glutamatergic or GABAergic tripartite synapses. Here, we utilized the Lattice structured illumination microscopy (Lattice SIM) to visualize Best1 expression at the perisynaptic junctions of the tripartite synapses in CA1 of mouse hippocampus. We performed co-labeling with antibodies against 1) Best1 and vesicular glutamate transporter-2 (vGLUT2) or 2) Best1 and vesicular GABA transporter (vGAT) to measure the proximity of Best1-containing perisynapse to glutamatergic or GABAergic presynapse, respectively. In addition, we examined two transgenic mouse lines of 1) APP/PS1 mouse showing high astrocytic MAOB activity and cytosolic GABA and 2) MAOB-KO mouse showing low astrocytic GABA. Lattice SIM images were further processed by Imaris, which allowed 3D-rendering and spot identification. We found that astrocytic Best1 was distributed closer to the glutamatergic synapses than GABAergic synapses in the wild-type mice. In APP/PS1 mice, Best1 distribution was significantly changed by moving away from the glutamatergic synapses while moving closer to the GABAergic synapses. On the contrary, in MAOB-KO mice, the Best1 distribution was dramatically changed by moving closer to the glutamatergic synapses and moving far away from the GABAergic synapses. Our findings propose that the proximity of Best1-containing perisynapses to presynapses dynamically changes according to the level of astrocytic cytosolic GABA.

Spore-forming Bacillus coagulans SNZ 1969 improved intestinal motility and constipation perception mediated by microbial alterations in healthy adults with mild intermittent constipation: A randomized controlled trial.

The spore-forming Bacillus coagulans has attracted attention for their therapeutic action in the colon. However, the mechanism of this action remains unclear. In this study, healthy subjects with mild intermittent constipation were supplemented with B. coagulans SNZ 1969 (BC) or the placebo for 8 weeks (n = 80). Then, we assessed colonic transit time (CTT), weekly complete spontaneous bowel movement (CSBM) scores, bowel discomfort symptom (BDS) scores, and 16S rRNA fecal microbiome profiles. The association between the critically altered gut microbiome and clinical outcomes was analyzed using redundancy analysis (RDA) and validated by receiver operating characteristic (ROC) curves. BC supplementation significantly improved CTT (p = 0.031), CSBM at weeks 2 (p = 0.045) and 9 (p = 0.038), and BDS at weeks 3 (p = 0.019) and 6 (p = 0.029) compared with the placebo, while altering the community composition of the gut microbiota. We also confirmed that BC was effectively delivered to the gut. Finally, the multivariate redundancy analysis concluded that BC-induced enrichment of Lactobacillales and diminishment of Synergistales were related to CTT improvements. This study provides important new data on how spore-forming B. coagulans SNZ 1969 contributes to improving gut motility and presents evidence supporting the use of B. coagulans SNZ 1969 in adults with mild intermittent constipation and habitual low intake of fruit and vegetables.

Bronchoalveolar lavage findings of radiation induced lung damage in rats.

Radiation induced lung damage is a main dose limiting factor when irradiating the thorax. Although Bronchoalveolar lavage (BAL) is a valuable tool for studying the mechanisms in pulmonary disorders, there are only a few studies about the BAL findings of radiation-induced lung damage. We evaluate the BAL findings for the evaluation of radiation-induced lung damage. Sprague-Dawley rats received 20 Gy of radiation to the right lung and control group were sham irradiated. BAL was performed for the right and left lungs separately 3, 7, 14, 28, and 56 days after radiation. The cells in the BAL fluid were counted and the concentrations of protein, NO, and TGF-beta in the BAL fluid were measured. Lung tissues were removed after BAL and stained with hematoxylin-eosin (H-E) and trichrome. From 2 weeks, histological findings showed definite lung damage. The protein level and TGF-beta in BAL fluid from the irradiated lung peaked at 4 and 8 weeks, respectively, after radiation. Total cell count in BAL fluid from both sides of lungs was increased from 2 weeks and continued to increase at 8 weeks after irradiation. NO in BAL fluid from both sides of lungs peaked at 4 weeks after irradiation. The protein level and TGF-beta were increased in BAL fluid from irradiated lungs. However, alveolar cells and NO increased in BAL fluid from both irradiated and non-irradiated lungs. BAL is a valuable tool for the evaluation of radiation induced lung damage.

Multivitamin and Mineral Supplementation Containing Phytonutrients Scavenges Reactive Oxygen Species in Healthy Subjects: A Randomized, Double-Blinded, Placebo-Controlled Trial.

Phytonutrients and vitamin and mineral supplementation have been reported to provide increased antioxidant capacity in humans; however, there is still controversy. In the current clinical trial, we examined the antioxidant and DNA protection capacity of a plant-based, multi-vitamin/mineral, and phytonutrient (PMP) supplementation in healthy adults who were habitually low in the consumption of fruits and vegetables. This study was an eight-week, double-blind, randomized, parallel-arm, and placebo-controlled trial. PMP supplementation for eight weeks reduced reactive oxygen species (ROS) and prevented DNA damage without altering endogenous antioxidant system. Plasma vitamins and phytonutrients were significantly correlated with ROS scavenging and DNA damage. In addition, gene expression analysis in PBMC showed subtle changes in superoxide metabolic processes. In this study, we showed that supplementation with a PMP significantly improved ROS scavenging activity and prevented DNA damage. However, additional research is still needed to further identify mechanisms of actions and the role of circulating phytonutrient metabolites.

Differential neutrophil infiltration contributes to regional differences in brain inflammation in the substantia nigra pars compacta and cortex.

Brain inflammation is a suggested risk factor for neurodegenerative disease. Interestingly, severe inflammation in the substantia nigra pars compacta (SNpc) accelerates the onset and progression of Parkinson's disease. In this study, we examined the underlying mechanisms of severe inflammation in the SNpc by comparing the inflammatory process with that in the cortex. In intact brain, the densities of CD11b(+) microglia were similar in the SNpc and cortex. However, lipopolysaccharide injection enhanced the CD11b(+) cell number in the SNpc, but not in the cortex. Previously, we reported that CD11b and myeloperoxidase (MPO) double-positive neutrophils infiltrate the SNpc following LPS injection (GLIA 55:1577-88). Notably, the MPO(+) neutrophil number increased dramatically in the SNpc, but only slightly in the cortex. The extent of neutrophil infiltration appeared to correlate with neuronal damage. We confirmed that loss of neurons in the SNpc was significantly reduced in neutropenic rats versus normal rats following LPS injection. In addition, the densities of astrocytes were much lower in the intact SNpc, compared with the cortex. Furthermore, after LPS injection, damage of endothelial cells and astrocytes, and blood-brain barrier (BBB) permeability was more pronounced in the SNpc. These results collectively suggest that excessive neutrophil infiltration and environmental factors, such as lower astrocyte density and higher BBB permeability, contribute to severe inflammation and neuronal death in the SNpc.

Low expression of Bax predicts poor prognosis in patients with locally advanced esophageal cancer treated with definitive chemoradiotherapy.

PURPOSE: The present study evaluated the prognostic significance of apoptosis-related proteins, p53, Bcl-2, Bax, and galectin-3 in patients with locally advanced esophageal cancer treated with definitive chemoradiotherapy. EXPERIMENTAL DESIGN: A total of 63 patients with locally advanced esophageal cancer (squamous cell carcinoma: 62; adenocarcinoma: 1; stages II-IV) were treated with definitive chemoradiotherapy using 5-fluorouracil and cisplatin combined with radiotherapy. Pretreatment tumor biopsy specimens were analyzed for p53, Bcl-2, Bax, and galectin-3 expression by immunohistochemistry. RESULTS: High expression of Bax, p53, Bcl-2, and galectin-3 was observed in 67%, 47%, 24%, and 29% of patients, respectively. The median overall survival (OS) of total patients was 14 months with 16% of 3-year OS. High expression of p53, Bcl-2, and galectin-3 did not show correlation with clinicopathologic characteristics, including patient outcome. Low expression of Bax was significantly correlated with lack of clinical complete response (P=0.023). Low expression of Bax was also associated with poor OS (median, 8 months versus 16 months; P=0.0008) in univariate analysis. In multivariate analysis, low expression of Bax was the most significant independent predictor of poor OS (P=0.009), followed by low dose intensity of cisplatin and lack of clinical complete response. CONCLUSIONS: Low expression of Bax was significantly associated with the poor survival of patients with locally advanced esophageal cancer treated with chemoradiotherapy using 5-fluorouracil and cisplatin. Immunohistochemical staining for Bax with a pretreatment biopsy specimen might be useful to select the optimal treatment options for these patients.

Astragali Radix elicits anti-inflammation via activation of MKP-1, concomitant with attenuation of p38 and Erk.

Although Astragali Radix (Astragalus, AR), the root of Astragalus membranaceus (Fisch) Bunge, is widely used in oriental medicine for tonifying the immune response and improving circulation, the underlying mechanism(s) by which these effects are induced remains unclear. Here, we report that AR displays anti-inflammatory effects in zymosan air-pouch mice by reducing the expression of iNOS, COX-2, IL-6, IL-1beta and TNF-alpha and by decreasing the production of nitric oxide (NO). In a similar manner, AR reduces the expression of IL-6, iNOS, and COX-2 in lipopolysaccharide (LPS)-treated Raw 264.7 cells. We further demonstrate that AR attenuates the activity of p38 and Erk1/2 and stimulates mitogen-activated protein kinase phosphatase-1 (MKP-1) in LPS-treated Raw 264.7 cells. Additionally, AR interferes with the translocation of NFkappaB to the nucleus, subsequently resulting in NFkappaB-dependent transcriptional repression. Taken together, these data reveal that AR has an anti-inflammatory effect that is mediated by the MKP-1-dependent inactivation of p38 and Erk1/2 and inhibition of NFkappaB-mediated transcription. These results imply that the AR herb has a potential anti-inflammatory activity.

Antitumor effect of 2-methoxyestradiol in a rat orthotopic brain tumor model.

Grade 4 malignant glioma (GBM) is a fatal disease despite aggressive surgical and adjuvant therapies. The hallmark of GBM tumors is the presence of pseudopalisading necrosis and microvascular proliferation. These tumor cells are hypoxic and express hypoxia-inducible factor-1 (HIF-1), a prosurvival transcription factor that promotes formation of neovasculature through activation of target genes, such as vascular endothelial growth factor. Here, we evaluated whether 2-methoxyestradiol, a microtubule and HIF-1 inhibitor, would have therapeutic potential for this disease in a 9L rat orthotopic gliosarcoma model using a combination of noninvasive imaging methods: magnetic resonance imaging to measure the tumor volume and bioluminescence imaging for HIF-1 activity. After imaging, histologic data were subsequently evaluated to elucidate the drug action mechanism in vivo. Treatment with 2-methoxyestradiol (60-600 mg/kg/d) resulted in a dose-dependent inhibition of tumor growth. This effect was also associated with improved tumor oxygenation as assessed by pimonidazole staining, decreased HIF-1alpha protein levels, and microtubule destabilization as assessed by deacetylation. Our results indicate that 2-methoxyestradiol may be a promising chemotherapeutic agent for the treatment of malignant gliomas, with significant growth inhibition. Further studies are needed to assess the effect of low or intermediate doses of 2-methoxyestradiol in combination with chemotherapeutic agents in clinical studies focused on malignant gliomas. In addition to showing tumor growth inhibition, we identified three potential surrogate biomarkers to determine the efficacy of 2-methoxyestradiol therapy: decreased HIF-1alpha levels, alpha-tubulin acetylation, and degree of hypoxia as determined by pimonidazole staining.

Effects of elm bark (Ulmus davidiana var. japonica) extracts on the modulation of immunocompetence in mice.

The immunomodulative effects of elm bark extract were studied in vitro by the proliferation of splenocytes and the production capacity of three kinds of cytokines [interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF)-alpha] by mouse peritoneal macrophages cultured with various fractions (methanol, hexane, chloroform, ethyl acetate, butanol, and water) of elm bark extract. Splenocyte proliferation and cell viability of peritoneal macrophages were increased with concentrations of polar fractions, such as butanol and water, in the range of 1-500 microg/mL. Significantly higher levels of the production of all three cytokines were detected with supplementation of methanol extract compared with other fractions. In order to elucidate its effect in vivo, elm bark water extract was orally administrated every other day for 2 weeks. Proliferation of splenocytes and the production capacity of cytokines (IL-1beta, IL-6, and TNF-alpha) by mouse peritoneal macrophages were used as indices for immune activity. Splenocyte proliferation induced by elm bark with lipopolysaccharide or concanavalin A stimulation was enhanced at 500 mg/kg of body weight concentrations compared to that of the control group. In the case of cytokines, the highest production of IL-6 and TNF was detected at 500 mg/kg of body weight concentrations. In conclusion, this study suggests through in vitro and in vivo experiments that Ulmus davidiana var. japonica (elm bark) extracts may enhance the immunocompetent properties such as splenocyte proliferation and cytokine production capacity by activated macrophages and have a protective effect in mice.

A novel regulatory element (E77) isolated from CHO-K1 genomic DNA enhances stable gene expression in Chinese hamster ovary cells.

Vectors flanked by regulatory DNA elements have been used to generate stable cell lines with high productivity and transgene stability; however, regulatory elements in Chinese hamster ovary (CHO) cells, which are the most widely used mammalian cells in biopharmaceutical production, are still poorly understood. We isolated a novel gene regulatory element from CHO-K1 cells, designated E77, which was found to enhance the stable expression of a transgene. A genomic library was constructed by combining CHO-K1 genomic DNA fragments with a CMV promoter-driven GFP expression vector, and the E77 element was isolated by screening. The incorporation of the E77 regulatory element resulted in the generation of an increased number of clones with high expression, thereby enhancing the expression level of the transgene in the stable transfectant cell pool. Interestingly, the E77 element was found to consist of two distinct fragments derived from different locations in the CHO genome shotgun sequence. High and stable transgene expression was obtained in transfected CHO cells by combining these fragments. Additionally, the function of E77 was found to be dependent on its site of insertion and specific orientation in the vector construct. Our findings demonstrate that stable gene expression mediated by the CMV promoter in CHO cells may be improved by the isolated novel gene regulatory element E77 identified in the present study.

Rectal bleeding and its management after irradiation for uterine cervical cancer.

PURPOSE: To present an evaluation of the risk factors of radiation-induced rectal bleeding and discuss its optimal management in patients with cancer of the cervix. METHODS AND MATERIALS: A total of 213 patients treated with radiotherapy (RT) alone were included. All patients underwent external beam radiotherapy with high-dose-rate brachytherapy. The rectal dose was calculated by both the International Commission on Radiation Units and Measurements (ICRU) Report 38 guidelines and the anterior rectal wall point on radiographs. Rectal bleeding was scored using the late effects normal tissue-subjective, objective, management, and analytic criteria. RESULTS: The incidence was 12.7% (27 of 213; Grade 1 in 9, Grade 2 in 16, and Grade 3 in 2). Most (92.6%) developed rectal bleeding within 2 years after RT completion (median 16 months). In univariate analysis, three significant factors were found: ICRU cumulative rectal biologically equivalent dose (CRBED) >100 Gy (19.7% vs. 4.2%), external beam RT dose to the parametrium >55 Gy (22.1% vs. 5.1%), and advanced stage (III-IV; 31.8% vs. 10.5%). In multivariate analysis, the ICRU-CRBED was the only significant factor. Six patients with Grade 1 bleeding experienced immediate relief with sucralfate enema for 1 month. For Grade 2 bleeding, sucralfate enema and/or coagulation were effective. Grade 3 bleeding lasted for 1 year despite frequent transfusions and coagulation. CONCLUSION: Grade 2 and 3 rectal bleeding occurred in 8.5% of patients. The most significant risk factor was the ICRU-CRBED. Prompt treatment with a combination of sucralfate enema and coagulation was effective in controlling Grade 1 and 2 rectal bleeding without the development of fistula or stricture.