Optical design and performance of the biological small-angle X-ray scattering beamline at the Taiwan Photon Source.

The optical design and performance of the recently opened 13A biological small-angle X-ray scattering (SAXS) beamline at the 3.0 GeV Taiwan Photon Source of the National Synchrotron Radiation Research Center are reported. The beamline is designed for studies of biological structures and kinetics in a wide range of length and time scales, from angstrom to micrometre and from microsecond to minutes. A 4 m IU24 undulator of the beamline provides high-flux X-rays in the energy range 4.0-23.0 keV. MoB4C double-multilayer and Si(111) double-crystal monochromators (DMM/DCM) are combined on the same rotating platform for a smooth rotation transition from a high-flux beam of ∼4 × 1014 photons s-1 to a high-energy-resolution beam of ΔE/E ≃ 1.5 × 10-4; both modes share a constant beam exit. With a set of Kirkpatrick-Baez (KB) mirrors, the X-ray beam is focused to the farthest SAXS detector position, 52 m from the source. A downstream four-bounce crystal collimator, comprising two sets of Si(311) double crystals arranged in a dispersive configuration, optionally collimate the DCM (vertically diffracted) beam in the horizontal direction for ultra-SAXS with a minimum scattering vector q down to 0.0004 Å-1, which allows resolving ordered d-spacing up to 1 µm. A microbeam, of 10-50 µm beam size, is tailored by a combined set of high-heat-load slits followed by micrometre-precision slits situated at the front-end 15.5 m position. The second set of KB mirrors then focus the beam to the 40 m sample position, with a demagnification ratio of ∼1.5. A detecting system comprising two in-vacuum X-ray pixel detectors is installed to perform synchronized small- and wide-angle X-ray scattering data collections. The observed beamline performance proves the feasibility of having compound features of high flux, microbeam and ultra-SAXS in one beamline.

Application of peripherally inserted central catheter in acute myeloid leukaemia patients undergoing induction chemotherapy.

Increasingly, peripherally inserted central catheters (PICC) are applied in patients with haematological malignancies. The feasibility and safety of PICC for induction chemotherapy in acute myeloid leukaemia (AML) remain unclear. Medical records of 89 newly diagnosed adult de novo AML patients, who achieved complete remission, were retrospectively reviewed (PICC group, n = 43; intravenous [IV] line group, n = 46). Patients' clinical characteristics and the number of blind punctures for blood sampling were compared between these two groups, and risk factors associated with bacteraemia were identified by univariate analysis. Patients in the PICC group experienced significantly fewer blind punctures than those in the IV line group (3.3 ± 3.6 vs. 14.4 ± 6.0; p = .000); 20.9% of PICC patients had bacteraemia, compared with 23.9% in the IV line group (p = .803). Most patients (76.7%) removed their PICC because treatment was completed. PICC increased the quality of life in AML patients undergoing chemotherapy induction by reducing the number of blind blood punctures required. Bacteraemia in PICC patients was comparable to that in IV line patients. PICC is, therefore, a feasible and safe central venous device for use in AML patients.

A home-based training programme improves family caregivers' oral care practices with stroke survivors: a randomized controlled trial.

OBJECTIVES: Stroke survivors experience poor oral health when discharged from the hospital to the community. The aim of this study was to evaluate the effectiveness of a home-based oral care training programme on knowledge, attitude, self-efficacy and practice behaviour of family caregivers. METHODS: A randomized controlled trial was conducted. The experimental group consisted of 48 family caregivers who received the home-based oral care training programme, and the control group consisted of 46 family caregivers who received routine oral care education. The outcomes were measured by the Knowledge of Oral Care, Attitude towards Oral Care, Self-Efficacy of Oral Care and Behaviour of Oral Care before the training programme, and at one and two months afterwards. The data were analysed using mixed model anova to determine differences in the outcomes between the two groups. RESULTS: The findings demonstrated that the intervention group had more knowledge (t = 8.80, P < 0. 001), greater self-efficacy (t = 3.53, P < 0.01) and better oral care behaviour (t = 11.93, P < 0.001) than the control group at one and two months, with statistically significant differences in oral care knowledge, self-efficacy and behaviour outcome over time. The attitude of the intervention group towards oral care practice was generally positive (mean of baseline and two month = 12.9 and 14.7), but no significant difference in attitude change between the control and intervention groups (t = 1.56, P = 0.12). The treatment interaction effect was significant for the family caregivers' behaviour of oral care at one and two months of the intervention for both groups. CONCLUSION: Our individualized home-based oral care education can achieve significant improvements in oral care knowledge and self-efficacy among family caregivers of stroke survivors, and it can sufficiently empower them to modify their oral care practices in a home-based healthcare environment.

Effect of family caregiver oral care training on stroke survivor oral and respiratory health in Taiwan: a randomised controlled trial.

OBJECTIVE: To evaluate the effectiveness of home-based oral care training programs on tongue coating (TC), dental plaque (DP), and symptoms of respiratory infection (SRI) in stroke survivors. METHODS: A single-blind, randomised, controlled trial conducted in a home-based setting over 2 months. Stroke survivors (n=48, experimental group) and their family caregivers received home-based oral care training programme while a control group of 46 stroke survivors and family caregivers received routine oral care education with swabs. TC, DP, and SRI were assessed at baseline and after one and two months, with results analysed using Mixed Model ANOVA. RESULTS: Poor oral hygiene and overall neglect of home oral care practices were observed at baseline. TC and DP scores were significantly reduced in the experimental group receiving the home-base oral care training program compared to the control group, who received only routine oral care education (P<0.001). The groupxtime interaction was significant, with decreased TC and DP scores for both groups at one month and at two months of additional care (when compared to baseline). The SRI scores were not significantly different between groups (P>0.05). The groupxtime interaction did not correlate with SRI for either group when compared to the baseline and to one month and two months of additional care. No adverse events were encountered and there was no external funding. CONCLUSIONS: Home-based oral care training programme had a beneficial effect on oral health as measured by TC and DP scores. The effect on SRI requires further longitudinal study.

Mutations in TWIST, a basic helix-loop-helix transcription factor, in Saethre-Chotzen syndrome.

Saethre-Chotzen syndrome is one of the most common autosomal dominant disorders of craniosynostosis in humans and is characterized by craniofacial and limb anomalies. The locus for Saethre-Chotzen syndrome maps to chromosome 7p21-p22. We have evaluated TWIST, a basic helix-loop-helix transcription factor, as a candidate gene for this condition because its expression pattern and mutant phenotypes in Drosophila and mouse are consistent with the Saethre-Chotzen phenotype. We mapped TWIST to human chromosome 7p21-p22 and mutational analysis reveals nonsense, missense, insertion and deletion mutations in patients. These mutations occur within the basic DNA binding, helix I and loop domains, or result in premature termination of the protein. Studies in Drosophila indicate that twist may affect the transcription of fibroblast growth factor receptors (FGFRs), another gene family implicated in human craniosynostosis. The emerging cascade of molecular components involved in craniofacial and limb development now includes TWIST, which may function as an upstream regulator of FGFRs.

Performance of the new biological small- and wide-angle X-ray scattering beamline 13A at the Taiwan Photon Source.

Recent developments in the instrumentation and data analysis of synchrotron small-angle X-ray scattering (SAXS) on biomolecules in solution have made biological SAXS (BioSAXS) a mature and popular tool in structural biology. This article reports on an advanced endstation developed at beamline 13A of the 3.0 GeV Taiwan Photon Source for biological small- and wide-angle X-ray scattering (SAXS-WAXS or SWAXS). The endstation features an in-vacuum SWAXS detection system comprising two mobile area detectors (Eiger X 9M/1M) and an online size-exclusion chromatography system incorporating several optical probes including a UV-Vis absorption spectrometer and refractometer. The instrumentation and automation allow simultaneous SAXS-WAXS data collection and data reduction for high-throughput biomolecular conformation and composition determinations. The performance of the endstation is illustrated with the SWAXS data collected for several model proteins in solution, covering a scattering vector magnitude q across three orders of magnitude. The crystal-model fittings to the data in the q range ∼0.005-2.0 Å-1 indicate high similarity of the solution structures of the proteins to their crystalline forms, except for some subtle hydration-dependent local details. These results open up new horizons of SWAXS in studying correlated local and global structures of biomolecules in solution.

Effects of suramin-related and other clinically therapeutic polyanions on protein kinase C activity.

The mechanism of the antineoplastic effects of suramin may involve interference with signal transduction, but in general is not well understood. We examined several polyanions to determine their effects on the kinase activity of the protein kinase C (PKC) beta1 and other PKC isoforms. Similar to suramin, a phosphorothioate oligodeoxynucleotide 28-mer homopolymer of cytidine (SdC28) inhibited the phosphatidylserine and Ca2+-dependent phosphorylation of an epidermal growth factor receptor octapeptide substrate. The inhibition by suramin was mixed competitive/noncompetitive with respect to ATP, but uncompetitive with respect to substrate. In contrast, the inhibition by SdC28 was competitive with respect to substrate (Ki = 5.4 microM) and not competitive with respect to ATP. The PKC alpha and beta1 isoforms were inhibited to the same extent with SdC28, while PKC epsilon was not inhibited. SdC28, in the absence of lipid cofactor, stimulated substrate phosphorylation, and in the absence of substrate induced PKC beta1 autophosphorylation. Similar behavior was seen with another polyanion, the polysulfated carbohydrate pentosan polysulfate (polyxylyl hydrogen sulfate). H4, a bis-naphthalene disulfonate tetraanion structurally related to suramin, also inhibited kinase activity but was not competitive with respect to ATP. Dianions closely related to H4 failed to inhibit PKC beta1, suggesting that multiple (>2) negative charges are required. The interactions of polyanions with PKC are complex, and are dependent on the molecular structure of the polyanion, the presence of cofactors, and the PKC isoform.

Ligand interactions with E-selectin. Identification of a new binding site for recognition of N-acyl aromatic glucosamine substituents of sialyl Lewis X.

Several N-acylglucosamine derivatives of sialyl Lewis X (1-3) were prepared using a combined chemical enzymatic approach and evaluated as an inhibitor of E-selectin-mediated cellular adhesion. Compounds with aromatic functionality, 1 and 2, were found to be 3-10 times more potent than the N-acetyl derivative (14) in an ELISA E-selectin cell adhesion assay. Conformational analysis with NMR indicated that the sialyl Lewis x domain of 1 retained the conformation of the N-acetyl derivative (14) despite the presence of the N-naphthamido group. The dramatic order of magnitude increase in potency of these monovalent structures can be utilized to design more potent selectin-based cell adhesion inhibitors.

Radionuclide demonstration of urinary extravasation with ureteral obstruction.

Two cases of urinary extravasation with ureteral obstruction demonstrated by the radionuclide studies are reported. The value of radionuclide studies in patients with renal transplantation has been reported previously, but studies in patients without transplantation have rarely been described in the literature. Ureteral obstruction may cause urinary extravasation, which may be demonstrated by radiouclide studies even when radiologic studies are inconclusive. In one case, urinary extravasation was detected in the sitting position but not in the supine position. Renal imaging should probably be performed not only with multiple projections but also in different positions.

A xanthine-based KMUP-1 with cyclic GMP enhancing and K(+) channels opening activities in rat aortic smooth muscle.

1. KMUP-1 (1, 3, 5 mg kg(-1), i.v.), a xanthine derivative, produced dose-dependent sustained hypotensive and short-acting bradycardiac effects in anaesthetized rats. This hypotensive effect was inhibited by pretreatment with glibenclamide (5 mg kg(-1), i.v.). 2. In endothelium-intact or denuded aortic rings preconstricted with phenylephrine, KMUP-1 caused a concentration-dependent relaxation. This relaxation was reduced by endothelium removal, the presence of NOS inhibitor L-NAME (100 microM) and sGC inhibitors methylene blue (10 microM) and ODQ (1 microM). 3. The vasorelaxant effects of KMUP-1 was attenuated by pretreatment with various K(+) channel blockers TEA (10 mM), glibenclamide (1 microM), 4-AP (100 microM), apamin (1 microM) and charybdotoxin (ChTX, 0.1 microM). 4. Increased extracellular potassium levels (30 - 80 mM) caused a concentration-related reduction of KMUP-1-induced vasorelaxations. Preincubation with KMUP-1 (1, 10, 100 nM) increased the ACh-induced maximal vasorelaxations mediated by endogenous NO release, and enhanced the potency of exogenous NO-donor SNP. 5. The vasorelaxant responses of KMUP-1 (0.01, 0.05, 0.1 microM) together with a PDE inhibitor IBMX (0.5 microM) had an additive action. Additionally, KMUP-1 (100 microM) affected cyclic GMP metabolism since it inhibited the activity of PDE in human platelets. 6. KMUP-1 induced a dose-related increase in intracellular cyclic GMP levels in rat A10 vascular smooth muscle (VSM) cells, but not cyclic AMP. The increase in cyclic GMP content of KMUP-1 (0.1 - 100 microM) was almost completely abolished in the presence of methylene blue (10 microM), ODQ (10 microM), and L-NAME (100 microM). 7. In conclusion, these results indicate that KMUP-1 possesses the following merits: (1) stimulation of NO/sGC/cyclic GMP pathway and subsequent elevation of cyclic GMP, (2) K(+) channels opening, and (3) inhibition of PDE or cyclic GMP breakdown. Increased cyclic GMP display a prominent role in KMUP-1-induced VSM relaxations.

Antiviral activity of Plantago major extracts and related compounds in vitro.

Plantago major L., a popular traditional Chinese medicine, has long been used for treating various diseases varying from cold to viral hepatitis. The aim of present study was to examine the antiviral activity of aqueous extract and pure compounds of P. major. Studies were conducted on a series of viruses, namely herpesviruses (HSV-1, HSV-2) and adenoviruses (ADV-3, ADV-8, ADV-11). The antiviral activity of EC50 was defined as the concentration achieved 50% cyto-protection against virus infection and the selectivity index (SI) was determined by the ratio of CC50 (concentration of 50% cellular cytotoxicity) to EC50. Results showed that aqueous extract of P. major possessed only a slight anti-herpes virus activity. In contrast, certain pure compounds belonging to the five different classes of chemicals found in extracts of this plant exhibited potent antiviral activity. Among them, caffeic acid exhibited the strongest activity against HSV-1 (EC50=15.3 microg/ml, SI=671), HSV-2 (EC50=87.3 microg/ml, SI=118) and ADV-3 (EC50=14.2 microg/ml, SI=727), whereas chlorogenic acid possessed the strongest anti-ADV-11 (EC50=13.3 microg/ml, SI=301) activity. The present study concludes that pure compounds of P. major, which possess antiviral activities are mainly derived from the phenolic compounds, especially caffeic acid. Its mode of action against HSV-2 and ADV-3 was found to be at multiplication stages (postinfection of HSV-1: 0-12 h; ADV-3: 0-2 h), and with SI values greater than 400, suggesting the potential use of this compound for treatment of the infection by these two viruses.

The expression of cytokines by an established basal cell carcinoma cell line (BCC-1/KMC) compared with cultured normal keratinocytes.

A basal cell carcinoma (BCC) cell line (BCC-1/KMC) has recently been successfully established from a patient. The production of interleukin-1 alpha (IL-1 alpha), IL-1 beta, IL-6 and IL-8 was assessed in comparison with that of cultured normal keratinocytes. The mRNA expression of these cytokines was measured by a reverse transcriptase-polymerase chain reaction (RT-PCR) method and the protein production by an ELISA. The cultured BCC cells spontaneously secreted more IL-6 and IL-8 but less IL-1 than the keratinocytes after culture for 24 h at 37 degrees C. It is suggested that the increased expression of IL-6 and IL-8 may indicate the transformation of normal keratinocytes to locally aggressive BCC.

Diuretic, natriuretic and hypotensive effects produced by Allium sativum (garlic) in anaesthetized dogs.

Gastric administration of encapsuled garlic powder to anaesthetized dogs induced dose-dependent (2.5 to 15 mg/kg) natriuretic and diuretic responses which reached maximum 30-40 min after garlic administration and decreased to basal levels after 100-150 min. A simultaneous decrease in arterial blood pressure was observed which continued past the 250 min-mark. High garlic doses (15 and 20 mg/kg) provoked bradycardia and T-wave inversion during the first 10-15 min of the experiment with recordings returning to normal and staying normal throughout the remainder of the experiment.

Inhibitory effect of garlic (Allium sativum) on sodium transport in isolated toad skin.

The effect of an aqueous fraction from the bulbs of Allium sativum (GE) was investigated in toad skin. When added to the inner (serosal) solution, GE caused a maximal reversible reduction of the transepithelial potential difference and short circuit current of 38% and 45%, respectively. When added to the outer (mucosal) solution, the effect was only partially reversible. Isaacson's amiloride test showed that GE decreased sodium potential (ENa.) and sodium conductance (GNa.). The net Na+ flux decreased due principally to a fall in Na+ flux in the active direction. GE decreased Na(+)-K+ ATPase activity in vitro. Partial replacement of sodium by choline in the outer solution reduced the effect of GE on the skin and substitution of normal Ringer's solution with isethionate Ringer's solution in the outer solution significantly enhanced the effect of GE on the skin. These results indicate that GE decreases active Na transport in the toad skin.

Characterization of an immortalized human cell line derived from neonatal foreskin diploid fibroblasts.

A new human skin cell line, designated as CCFS-1/KMC, immortalized from human neonatal foreskin diploid fibroblast cells, has been subcultured successfully in vitro for more than 500 passages. This anchorage-dependent cell line possesses many common features of transformation such as morphological and cytoskeletal changes, hypotriploidy, infinite lifespan, increasing plating efficiency and saturation density, and decreasing serum requirement and population doubling time. Human papillomavirus (HPV) type 18 DNA was detected in the cell line before and after immortalization by the polymerase chain reaction (PCR) method. Tumorigenicity, however, was not demonstrated in vivo. The isoenzyme activity of the cell line shows activation of a placental form of alkaline phosphatase and a changing lactate dehydrogenase isoenzyme pattern that is different from transformation by carcinogens. Class I HLA and class II HLA antigens are constitutively expressed on this skin cell line. Here we report that these immortalized human fibroblasts derived from neonatal HPV-18-DNA-contained diploid fibroblasts possess double minute chromosomes (DMs), a karyotypic aberration usually found in cancer cells.

Effects of VEGF121 and/or VEGF165 gene transfection on collateral circulation development.

BACKGROUND AND PURPOSE: Angiogenesis is regulated by various factors, including vascular endothelial growth factor (VEGF). Five isoforms of VEGF have been discovered: VEGF121, VEGF145, VEGF165, VEGF189, and VEGF206. The teleologic basis for the various VEGF isoforms remains unclear, but different VEGF isoforms may mediate distinct endothelial cell functions such as angiogenesis, vascular permeability, and differentiation. We sought to determine the effects of various VEGF isoforms on angiogenesis under ischemic conditions in rabbits. METHODS: The effects of VEGF121 and/or VEGF165 gene transfection on collateral circulation development in ischemic rabbit hindlimb muscles were investigated by using naked plasmids encoding VEGF121 or VEGF165 (pVEGF121 or pVEGF165), either individually or in combination. pCMV beta was used as the control plasmid. The femoral artery on one side of New Zealand White rabbits was ligated. Ten days later, the ischemic muscles received direct intramuscular injection of pVEGF121 (500 micrograms), pVEGF165 (500 micrograms), or pVEGF121 (250 micrograms) + pVEGF165 (250 micrograms) in experimental groups, while pCMV beta (500 micrograms) was used in the control group. Therapeutic effects were evaluated 30 days later by anatomic and physiologic analysis. RESULTS: Internal iliac angiography showed strong development of collateral circulation in all of the pVEGF-treated groups. In contrast, collateral arteries developed weakly in the control group. Combination treatment with both pVEGF121 and pVEGF165 did not result in additional improvement compared with pVEGF121 or pVEGF165 treatment alone (angiographic scores: pVEGF121 = 0.85 +/- 0.10; pVEGF165 = 0.81 +/- 0.11; pVEGF121 + pVEGF165 = 0.83 +/- 0.09; control = 0.53 +/- 0.09; p < 0.01). A favorable response in the development of circulation at the capillary level with pVEGF121 and/or pVEGF165 versus pCMV beta was also found. Blood pressure measurement and regional blood flow measurement using colored microspheres revealed similar results. CONCLUSIONS: Our results show that direct intramuscular injection of naked DNA encoding VEGF121 or VEGF165, individually or in combination, is an effective method for gene transfer in an animal model of ischemic limbs and results in augmented collateral vascular development and tissue perfusion.

Antileukemic activity of Bidens pilosa L. var. minor (Blume) Sherff and Houttuynia cordata Thunb.

To evaluate the anti-leukemic activity of Bidens pilosa L. var. minor (Blume) Sherff and Houttuynia cordata Thunb., cytotoxicity tests with an XTT-based colorimetric assay were used. Five leukemic cell lines, namely L1210, U937, K562, Raji and P3HR1, were cultured with hot water extracts of B. pilosa var. minor or H. cordata. Hot water extracts of B. pilosa var. minor inhibited these five leukemic cells with IC50s between 145 microg/ml and 586 microg/ml. The effect was greatest on four cell lines, namely L1210, P3MR1, Raji and K562, with IC50s below 200 microg/ml and a selective index of more than 5. Hot water extract of H. cordata inhibited these five leukemic cells with IC50s between 478 microg/ml and 662 microg/ml. The selective index was between 1.5 and 2.1. B. pilosa var. minor was more effective than H. cordata in inhibiting most of the leukemic cells in our study. We suggest that B. pilosa L. var. minor (Blume) Sherff may prove to be a useful medicinal plant for treating leukemia.

Characterization of a new human transitional cell carcinoma cell line from the renal pelvis, RTCC-1/KMC.

Tumors of the renal pelvis and ureter are relatively common malignancies in Taiwan. Studying the genetic or biochemical aberrations is a feasible pursuit that has great potential to further our understanding of urothelial cancer and may provide clinically valuable information. We now report a new long-term culture (RTCC-1/KMC) of human TCC derived from the renal pelvis, which is aimed to be used as a target for those studying in this field. The cultured cells exhibited anchorage independence and loss of contact inhibition. Chromosomal analysis revealed an aneuploidy line with a modal number of 50. Population doubling time was about 36 hours at the third passage. Expression of keratin proteins confirmed its epithelial origin. The genetic markers of the RTCC -1/KMC cell line were HLA-A11, B46, B60, Cw1, Cw7, DRw12 and DRw16. The human papillomavirus and herpes simplex virus genomes were not found in this cell line.