Terpenoids with anti-influenza activity from the leaves of Euphorbia leucocephala.

Two rearranged terpenoids with a rare 3,4,5-trimethyl-cyclohexa-2,5-dien-1-one moiety, namely leucocephins A (1) and B (2), and a megastigmane, namely leucocephin C (3), as well as three known compounds, hollongdione (4), 3-acetoxy-lup-12,20(29)-diene (5), and ß-amyrin acetate (6) were isolated from the leaves of Euphorbia leucocephala. Their structures and absolute configurations were determined by spectroscopic methods and comparing with literature data. Compounds 4-6 exhibited potent anti-influenza A virus activity comparable to the positive control, betulinic acid.

Anti-Mouse CD83 Monoclonal Antibody Targeting Mature Dendritic Cells Provides Protection Against Collagen Induced Arthritis.

Antibodies targeting the activation marker CD83 can achieve immune suppression by targeting antigen-presenting mature dendritic cells (DC). This study investigated the immunosuppressive mechanisms of anti-CD83 antibody treatment in mice and tested its efficacy in a model of autoimmune rheumatoid arthritis. A rat anti-mouse CD83 IgG2a monoclonal antibody, DCR-5, was developed and functionally tested in mixed leukocyte reactions, demonstrating depletion of CD83+ conventional (c)DC, induction of regulatory DC (DCreg), and suppression of allogeneic T cell proliferation. DCR-5 injection into mice caused partial splenic cDC depletion for 2-4 days (mostly CD8+ and CD83+ cDC affected) with a concomitant increase in DCreg and regulatory T cells (Treg). Mice with collagen induced arthritis (CIA) treated with 2 or 6 mg/kg DCR-5 at baseline and every three days thereafter until euthanasia at day 36 exhibited significantly reduced arthritic paw scores and joint pathology compared to isotype control or untreated mice. While both doses reduced anti-collagen antibodies, only 6 mg/kg achieved significance. Treatment with 10 mg/kg DCR-5 was ineffective. Immunohistological staining of spleens at the end of CIA model with CD11c, CD83, and FoxP3 showed greater DC depletion and Treg induction in 6 mg/kg compared to 10 mg/kg DCR-5 treated mice. In conclusion, DCR-5 conferred protection from arthritis by targeting CD83, resulting in selective depletion of mature cDC and subsequent increases in DCreg and Treg. This highlights the potential for anti-CD83 antibodies as a targeted therapy for autoimmune diseases.

Abdominal Ultrasound in the Detection of an Incidental Paraganglioma.

Paraganglioma is a tumor that originates from neuroendocrine cells of the sympathetic or parasympathetic systems. Patients may suffer from headaches, palpitations, diaphoresis, and hypertension due to catecholamine excess or symptoms from the mass effect of the tumor. In the absence of typical symptoms of catecholamine excess, the diagnosis of a nonfunctional paraganglioma is often delayed. Herein, we report a case of a 63-year-old female patient with a nonfunctional paraganglioma which is an accidental finding during investigation of a fever. Abdominal ultrasonography incidentally detected this lesion as a complex, solid, cystic mass in the left suprarenal retroperitoneum.

Dihydrotestosterone (DHT) Enhances Wound Healing of Major Burn Injury by Accelerating Resolution of Inflammation in Mice.

Androgens have been known to inhibit cutaneous wound healing in men and male mice. However, in children with major burn injuries, a synthetic androgen was reported clinically to improve wound healing. The aim of this study is to investigate the role of dihydrotestosterone (DHT) as a new therapeutic approach in treating major burn injury. In the present study, mice received systemic androgen treatment post major burn injury. Wound healing rate and body weight were monitored over 21 days. The serum level of inflammatory cytokines/chemokines were measured using multiplex immunoassays. In addition, splenocyte enumeration was performed by flow cytometry. Healing phases of inflammation, re-epithelialization, cell proliferation and collagen deposition were also examined. In results, DHT treated mice lost less weight and displayed accelerated wound healing but has no impact on hypermetabolism. Mice, after burn injury, displayed acute systemic inflammatory responses over 21 days. DHT treatment shortened the systemic inflammatory response with reduced splenic weight and monocyte numbers on day 14 and 21. DHT treatment also reduced wound infiltrating macrophage numbers. In conclusion, DHT treatment facilitates local wound healing by accelerating the resolution of inflammation, but not through alterations of post-burn hypermetabolic response.

Standardisation of flow cytometry for whole blood immunophenotyping of islet transplant and transplant clinical trial recipients.

Understanding the immunological phenotype of transplant recipients is important to improve outcomes and develop new therapies. Immunophenotyping of whole peripheral blood (WPB) by flow cytometry is a rapid method to obtain large amounts of data relating to the outcomes of different transplant treatments with limited patient impact. Healthy individuals and patients with type 1 diabetes (T1D) enrolled in islet transplantation were recruited and WPB was collected. 46 fluorochrome-conjugated mouse-anti-human antibodies were used (43 of 46 antibodies were titrated). BD cytometer setup and tracking beads were used to characterize and adjust for cytometer performance. Antibody cocktails were pre-mixed <60 minutes before staining. Multicolour panels were designed based on fluorochrome brightness, antigen density, co-expression, and fluorochrome spillover into non-primary detectors in each panel on a 5 laser flow cytometer. WPB sample staining used 50-300 µl WPB for each panel and was performed within 2 hours of blood sample collection. Samples were acquired on a BD-LSRFortessa. The operating procedures, including specimen collection, antibody cocktails, staining protocol, flow-cytometer setup and data analysis, were standardized. The staining index of 43 antibodies and the spillover spreading matrix for each panel was calculated. The final concentrations for the 46 antibodies used was determined for staining of WPB samples. Absolute cell-count and 7 leukocyte profiling panels consisting of subsets and/or status of granulocytes, monocytes, dendritic, B, NK, and T cells including regulatory T cells (Tregs) and NKT were designed and established on a 5 laser BD-LSR Fortessa. 13 T1D patients, including 4 islet transplant recipients and 8 healthy controls, were evaluated. The ability to reproducibly measure immune subsets and immune-profiles of islet transplant patients up to 18 months post transplantation has been established as a tool to measure immune cell reconstitution after transplantation.

Protein-DNA complex-guided discovery of the antibacterial lead E1 for restoring the susceptibility of Klebsiella Pneumoniae to polymyxin B by targeting the response regulator PmrA.

A new antibacterial drug is urgently needed. We employed a protein-DNA complex-guided pharmacophore modeling approach to screen inhibitors against the response regulator PmrA of polymyxin B-resistant Klebsiella pneumoniae (KP). The identified lead, E1 (IC50 = 10.2 µM), targeted the DNA-binding domain of PmrA (KD = 1.7 µM), whose conserved residues R171, R198, K203, and Y214 have been shown to be hotspots for antimicrobial development. Treatment of E1 restored the susceptibility of KP to polymyxin B.

Novel augmentation by bufalin of protein kinase C-induced cyclooxygenase-2 and IL-8 production in human breast cancer cells.

Cyclooxygenase-2 (COX-2) and IL-8 are two inflammatory mediators induced by protein kinase C (PKC) via various stimuli. Both contribute significantly to cancer progression. Bufalin, a major active component of the traditional Chinese medicine Chan Su, is known to induce apoptosis in various cancer cells. This study clarifies the role and mechanism of bufalin action during PKC regulation of COX-2/IL-8 expression and investigates the associated impact on breast cancer. Using MB-231 breast cancer cells, bufalin augments PKC induction of COX-2/IL-8 at both the protein and mRNA levels, and the production of prostaglandin E2 (PGE2) and IL-8. The MAPK and NF-κB pathways are involved in both the PKC-mediated and bufalin-promoted PKC regulation of COX-2/IL-8 production. Bufalin increases PKC-induced MAPKs phosphorylation and NF-κB nuclear translocation. PGE2 stimulates the proliferation/migration of breast cancer cells. Furthermore, PKC-induced matrix metalloproteinase 3 expression is enhanced by bufalin. Bufalin significantly enhances breast cancer xenograft growth, which is accompanied by an elevation in COX-2/IL-8 expression. In conclusion, bufalin seems to promote the inflammatory response in vitro and in vivo, and this occurs, at least in part, by targeting the MAPK and NF-κB pathways, which then enhances the growth of breast cancer cells.

Antibacterial and Synergistic Activity of Pentacyclic Triterpenoids Isolated from Alstonia scholaris.

(1) BACKGROUND: Alstonia scholaris (Apocynaceae) is an important medicinal plant that has been historically used in "Dai" ethnopharmacy to treat infectious diseases in China. Although various pharmacological activities have been reported, the antimicrobial constitutes of A. scholaris have not yet been identified. The objective of this study is to evaluate the antibacterial constitutes from the leaf extract of A. scholaris and to assess the synergistic effects of isolated compounds with antibiotics against bacterial pathogens.; (2) METHODS: The chemical constitutes isolated from the leaf extract of A. scholaris were structurally identified by NMR. The antibacterial and synergistic effect of compounds was assessed by calculating the minimal inhibitory concentration (MIC), checkerboard dilution test, and time-kill assay.; (3) RESULTS: Six pentacyclic triterpenoids were structurally identified as (1) lupeol, (2) betulin, (3) 3-hydroxy-11-ursen-28,13-olide, (4) betulinic acid, (5) oleanolic acid and (6) ursolic acid. Both oleanolic and ursolic acid showed antibacterial activity but were limited to Gram-positive bacteria. Ursolic acid showed a synergistic effect with ampicillin and tetracycline against both Bacillus cereus and S. aureus.; (4) CONCLUSION: These findings reflect that pentacyclic triterpenoids are the antibacterial chemicals in A. scholaris. The ability of ursolic acid to enhance the activity of antibiotics can constitute a valuable group of therapeutic agents in the future.

A CT-scan database for the facial soft tissue thickness of Taiwan adults.

Facial reconstruction is a branch of forensic anthropology used to assist in the identification of skeletal remains. The majority of facial reconstruction techniques use facial soft tissue depth chart data to recreate facial tissue on a skull or a model of a skull through the use of modeling clay. This study relied on 193 subjects selected from the Taiwanese population on the basis of age and gender to determine the average values of 32 landmarks, include midline and bilateral measures, by means of CT scans. The mean age of the subjects was 46.9±16.4 years, with a mean age of 43.8±16.6 for males and 49.9±15.8 for females respectively. There were 16 landmarks with statistically significant differences between male and female subjects, namely S, G, N, Na, Ph, Sd and Id in the midline portion, FE, LO, ZA and Sub M2 in the bilateral-right and left portion, and IM point in the bilateral-left portion (abbreviations adapted from Karen T. Taylor's work). The mean soft tissue depth was greater in males than in females, and there was significant difference between the right and left sides of the face in Za point. This study's findings were compared with those of Bulut et al.

The role of pentacyclic triterpenoids in the allelopathic effects of Alstonia scholaris.

Alstonia scholaris is a tropical evergreen tree native to South and Southeast Asia. Alstonia forests frequently lack understory species. However, potential mechanisms-particularly the allelochemicals involved-remain unclear. In the present study, we identified allelochemicals of A. scholaris, and clarified the role of allelopathic substances from A. scholaris in interactions with neighboring plants. We showed that the leaves, litter, and soil from A. scholaris inhibited growth of Bidens pilosa-a weed found growing abundantly near A. scholaris forests. The allelochemicals were identified as pentacyclic triterpenoids, including betulinic acid, oleanolic acid, and ursolic acid by using (1)H and (13)C-NMR spectroscopy. The half-maximal inhibitory concentration (IC50) for radicle growth of B. pilosa and Lactuca sativa ranged from 78.8 µM to 735.2 µM, and ursolic acid inhibited seed germination of B. pilosa. The triterpenoid concentrations in the leaves, litter, and soil were quantified with liquid chromatography-electrospray ionization/tandem mass spectrometry. Ursolic acid was present in forest soil at a concentration of 3,095 µg/g, i.e., exceeding the IC50. In the field, ursolic acid accumulated abundantly in the soil in A. scholaris forests, and suppressed weed growth during summer and winter. Our results indicate that A. scholaris pentacyclic triterpenoids influence the growth of neighboring weeds by inhibiting seed germination, radicle growth, and functioning of photosystem II.

Modeling ozone mass transfer in reclaimed wastewater.

Ozone mass transfer in reclaimed water was evaluated at pilot scale to determine mass-transfer characteristics and reaction kinetics and to assess the use of oxygen as a surrogate to measure this process. Tests were conducted in a 40-L/min pilot plant over a 3-year period. Nonsteady-state mass-transfer analyses for both oxygen and ozone were performed for superficial gas flow rates ranging from 0.13m/min to 0.40m/min. The psi factor, which is the ratio of volumetric mass-transfer coefficients of ozone to oxygen, was determined. The decrease in oxygen transfer rate caused by contaminants in reclaimed water was only 10 to 15% compared to tap water. A simple mathematical model was developed to describe transfer rate and steady state ozone concentration. Ozone decay was modeled accurately as a pseudo first-order reaction between ozone and ozone-demanding materials.

Enhancement of cellulose pellicle production by constitutively expressing vitreoscilla hemoglobin in Acetobacter xylinum.

Vitreoscilla hemoglobin (VHb) gene driven by the constitutive bla promoter was expressed in the cellulose-producing Acetobacter xylinum. The expressed VHb was biochemically active and could enhance cell growth in a shaken culture containing cellulase. VHb-expressing A. xylinum (VHb+) exhibited a specific growth rate 50% higher than that of the host strain (VHb-). Probably because of its faster growth rate, the size of tentacled cellulose beads produced by VHb+ was about 20% of that produced by VHb- after 2 days cultivation in a shake-flask. When cultured statically, the amount of cellulose pellicle produced by VHb+ could be 2-fold that produced by VHb-. Cellulose pellicle concentration of 11 g/L was obtained for VHb+, whereas 6 g/L was obtained for VHb- after 6 days of microaerobic incubation.

Toxic keratopathy associated with abuse of low-dose anesthetic: a case report.

OBJECTIVE: To describe the clinical course and treatment of toxic keratopathy associated with abuse of topical anesthetic at a very low concentration, 0.05%. METHOD: Case report. RESULT: A 47-year-old female systemic lupus erythematosus (SLE) patient with blurred vision and irritated right eye was referred to the ophthalmology department of our hospital. Under slit-lamp microscope, a 5.5 x 4.5 mm central corneal epithelial defect with underlying infiltrative and opaque stroma was noted in her right eye. Two weeks before, a corneal ulcer was diagnosed, and oxybuprocaine 0.05% (Lacrimin, Santen, Osaka, Japan) eye drops were prescribed 4 times daily but used every 5 to 10 minutes because the right eye was severely irritated. She was admitted immediately under the impression of toxic corneal ulcer. Preservative-free lubricants and prophylactic topical antibiotics 4 times daily were applied. Therapeutic soft contact lens was started after no infective agents were detected. Two weeks later, the stromal infiltration subsided, and the corneal epithelium was slowly healing, but superficial punctate epithelial defects at the lesion site persisted for another 6 months. The vision of her right eye improved from finger-counting at a 30-cm distance to 20/1200 with correction. CONCLUSION: Toxic keratopathy may result from abuse of topically administered anesthetics even at a very low concentration, 0.05%. Because this SLE patient has tear problems, we suggest that topical anesthetics must be used very cautiously and never prescribed to patients with dry eyes where the integrity of ocular surface is altered.

Design and implementation of a real-time clinical alerting system for intensive care unit.

Nowadays many hospitals use clinical information systems (CIS) to improve patient health care and information management. CIS retrieves and manages patient information such as patient administration, laboratory data and medications. Specialized CIS, especially Intensive Care Unit (ICU) CIS, needs to handle more additional information. How to process and make use of this enormous amount of physiologic data generated by ICU is a significant topic in CIS design. We have designed and implemented a real-time clinical alerting system for ICU, which executes alert algorithms on the database records of 63 ICU physiologic parameters. The user can create various alert rules with our rule editor to supervise the values or trends of these parameters. When an alert condition on a specific patient is detected, the system notices the clinicians in charge with mobile phones or pagers. With this functionality, up-to-date patient information is provided for clinicians inverted exclamation mark | use in judging the patient inverted exclamation mark |s condition and making treatment decisions.