Predictors of general discomfort, limitations in activities of daily living and intention of a second donation in unrelated hematopoietic stem cell donation.

We performed a retrospective study of 1868 consecutive unrelated donors to predict the risk factors related to general discomfort, limitations in activities of daily living (ADLs) and intention of a second donation in hematopoietic stem cell (HSC) donation. General discomfort and limitations in ADLs were assessed by numerical measurement (scores of 0-10) and donor's intention of a second donation by yes or no reply. The post-donation questionnaires were completed within 48 h after HSC collection and at 1 week, 4 weeks, and 4 months thereafter. Predictors of general discomfort included female sex (P<0.0001), bone marrow (BM) collection (P<0.0001) or PBSC collection through a central line (CL; P=0.0349), 2-day collection (P=0.0150) and negative or undetermined intention of a second donation on day 1 (P<0.0001). Predictors of limitations in ADLs included age group of 30-39 years (P=0.0046), female sex (P<0.0001), BM collection (P<0.0001) or PBSC collection through a CL (P<0.0001) and negative or undetermined intention of a second donation on day 1 (P<0.0001). The only predictor of positive intention of a second donation was male sex (P=0.0007). Age, sex and collection method and period should be considered risk factors when unrelated HSC donation is performed.

[Heterologous allogeneic blood transfusion as a risk factor associated with the adverse course of the surgical wound in primary hip surgery]. / Hemotransfusión alogénica heteróloga como factor de riesgo asociado con la evolución adversa de la herida quirúrgica en cirugía primaria de cadera.

BACKGROUND: Due to its immunomodulating effects mediated by leukocytes and interleukins, heterologous allogeneic blood transfusion has been considered as a risk factor for both morbidity and mortality in patients undergoing orthopedic surgery, including hip surgery. This research analyzed whether heterologous allogeneic blood transfusion is a risk factor associated with the adverse course or complication of the surgical wound in patients undergoing primary hip surgery due to fracture at a general hospital in 2008-2009. MATERIAL AND METHODS: Forty-nine patients who had a complication (cases) and 207 with no complications (controls) were identified and both groups were compared with a bivariate and multivariate analysis, and demographic and clinical data, including having undergone blood transfusion or not. RESULTS: Not having received a blood transfusion was identified as a variable that reduced the risk of surgical wound complications (OR = 0.05, 95% confidence interval [CI 95%] 0.0067 to 0.16; chi2 with p < 0.001). The multivariate model excluded as clinically significant variables the duration of surgery (OR = 1.01, CI 95% 0.99 to 1.02; p = 0.12) and certain chronic conditions (OR = 0.54, CI 95% 0.13 to 2.24 for diabetes mellitus, OR = 1.16, CI 95% 0.29 to 4.60 for chronic hypertension, OR = 1.21, CI 95% 0.19 to 7.51 for various heart diseases). CONCLUSIONS: Not having received a blood transfusion reduced 95% the risk of surgi- cal wound complications. Neither the duration of surgery nor a specific comorbid condition were associated with the former event.

Maintenance and characterization of multipotent mesenchymal stem cells isolated from canine umbilical cord matrix by collagenase digestion.

Multipotent mesenchymal stem cells have been considered as a novel clinical approach for cell therapy and regenerative medicine. In this study, mesenchymal stem cells (MSCs) were successfully isolated from canine umbilical cord matrix (cUCM; also referred to as Wharton's Jelly) by collagenase digestion and further characterized for multipotent properties associated with MSCs. Our cUCM-derived MSCs (cUCM-MSCs) were plastic adherent, spindle-shaped and fibroblast-like cells, maintaining expression of pluripotency markers such as Oct3/4, Nanog, Sox-2 and SSEA-4 as well as normal chromosomal number during a long-term proliferative culture. The cells expressed MSCs-specific surface markers, including CD44, CD90, CD105, and CD184, but did not CD29, CD33, CD34, and CD45. More importantly, cUCM-MSCs could differentiate into mesodermal (adipocyte, osteocyte and chondrocyte) and ectodermal (neuronal cell) cell lineages. These results imply that collagenase digestion would be a highly effective way to isolate multipotent MSCs in abundant amounts.

Implementation and performance evaluation of mobile ad hoc network for Emergency Telemedicine System in disaster areas.

So far we have developed Emergency Telemedicine System (ETS) which is a robust system using heterogeneous networks. In disaster areas, however, ETS cannot be used if the primary network channel is disabled due to damages on the network infrastructures. Thus we designed network management software for disaster communication network by combination of Mobile Ad hoc Network (MANET) and Wireless LAN (WLAN). This software maintains routes to a Backbone Gateway Node in dynamic network topologies. In this paper, we introduce the proposed disaster communication network with management software, and evaluate its performance using ETS between Medical Center and simulated disaster areas. We also present the results of network performance analysis which identifies the possibility of actual Telemedicine Service in disaster areas via MANET and mobile network (e.g. HSDPA, WiBro).

Galactosylated chitosan-graft-polyethylenimine as a gene carrier for hepatocyte targeting.

Chitosans have been proposed as alternative, biocompatible cationic polymers for nonviral gene delivery. However, the low transfection efficiency and low specificity of chitosan need to be addressed before clinical application. We prepared galactosylated chitosan-graft-polyethylenimine (GC-g-PEI) copolymer by an imine reaction between periodate-oxidized GC and low-molecular-weight PEI. The molecular weight and composition were characterized using gel permeation chromatography column with multi-angle laser scattering and (1)H nuclear magnetic resonance, respectively. The copolymer was complexed with plasmid DNA in various copolymer/DNA (N/P) charge ratios, and the complexes were characterized. GC-g-PEI showed good DNA-binding ability and superior protection of DNA from nuclease attack and had low cytotoxicity compared to PEI 25K. GC-g-PEI/DNA complexes showed higher transfection efficiency than PEI 25K in both HepG2 and HeLa cell lines. Transfection efficiency into HepG2, which has asialoglycoprotein receptors, was higher than that into HeLa, which does not. GC-g-PEI/DNA complexes also transfected liver cells in vivo after intraperitoneal (i.p.) administration more effectively than PEI 25K. These results suggest that GC-g-PEI can be used in gene therapy to improve transfection efficiency and hepatocyte specificity in vitro and in vivo.

Receptor-mediated gene delivery using chemically modified chitosan.

Chitosan has been investigated as a non-viral vector because it has several advantages such as biocompatibility, biodegradability and low toxicity with high cationic potential. However, the low specificity and low transfection efficiency of chitosan need to be solved prior to clinical application. In this paper, we focused on the galactose or mannose ligand modification of chitosan for enhancement of cell specificity and transfection efficiency via receptor-mediated endocytosis in vitro and in vivo.

Partial characterization of farnesyl and geranylgeranyl diphosphatases induced in rice seedlings by UV-C irradiation.

This study reports the identification and characterization of allyl diphosphatases (EC 3.1.7.1) in plants by using rice seedlings. Two distinct Mg(2+)-independent allyl diphosphatases, which convert farnesyl diphosphate (FDP) and geranylgeranyl diphosphate (GGDP) into farnesol and geranylgeraniol, respectively, were induced in rice seedlings irradiated with UV-C. Farnesyl diphosphatase (FDPase) and geranylgeranyl diphosphatase (GGDPase) are located in the microsomal fraction. The relative specific activity of FDPase was much higher than the specific activity of GGDPase. FDPase activity was inhibited by GGDP (50% inhibition at 5 microM) and geranyl diphosphate (50% inhibition at 100 microM). In contrast, GGDPase activity was inhibited 50% by 50 microM isopentenyl diphosphate or 100 microM FDP. The optimal pH for FDPase was 6.3 and for GGDPase was 7.9.

Water-soluble and low molecular weight chitosan-based plasmid DNA delivery.

PURPOSE: Chitosan, a natural cationic polysaccharide, is a candidate non-viral vector for gene delivery because of its high positive charges and low cytotoxicity. In this study, low molecular weight chitosan (LMWC, molecular weight of 22 kDa) was characterized and evaluated as a gene carrier. METHODS: Plasmid/LMWC complex was analyzed in 1% agarose gel electrophoresis. To confirm that the LMWC protected plasmids from nuclease. DNase I protection assays were performed. pSV-beta-galactosidase plasmid/LMWC complex was transfected into 293T cells and transfection efficiency was evaluated by beta-galactosidase assay. Cytotoxicity of LMWC was determined by MTT assay. RESULTS: Unlike high molecular weight chitosan (HMWC), LMWC is highly water soluble, and can form complex with plasmids in physiological buffer. The plasmid DNA was completely retarded at a weight ratio of 1:2 (plasmid:LMWC) in 1% agarose gel. DNase I protection assay showed that plasmids were protected from DNase-I over 60 min. The most efficient transfection was obtained at a weight ratio of 1:3 (plasmid:LMWC). The transfection efficiency of LMWC was significantly higher than naked DNA and higher than poly-L-lysine (PLL). MTT assay showed that LMWC was less cytotoxic than PLL. CONCLUSIONS: LMWC is non-toxic and has higher transfection efficiency than PLL. Therefore, LMWC will be useful in the development of safe gene carriers.

[Epidemiologic features of tetanus in adults form Yucatan, Mexico]. / Características epidemiológicas del tétanos en adultos en Yucatán, México.

OBJECTIVE: To describe epidemiologic features from tetanus in adult patients (TIAP) treated at hospital general O'Horan in Merida, Yucatan, Mexico and compare them with another Mexican series analyzed 25 years ago. MATERIAL AND METHODS: From 1985 to 1999, 121 TIAP cases aged 13 years or older were identified from which 112 were analyzed. Diagnosis of TIAP was made just in a clinical basis. To analyze information inferencial statistics were used. RESULTS: People affected by tetanus averaged 43 +/- 21, 95% CI 39 to 47. A 3:1 male to female ratio was documented. Eighty two (73%) patients come from the rural area; 43 (38%) did work as peasants. Sixty two cases (55%) were diagnosed during the Fall and Winter seasons. In 91 patients (81%) no anti-tetanus vaccination was documented. In 89 cases (79%) incubation period averaged 5.4 +/- 4 days, 95% CI 5 to 6. According to this 89 cases (79%) with incubation period < 10 days were graded as severe tetanus and 23 (21%) with incubation period > or = 10 days were graded as non-severe tetanus. Tetanus-prone wounds were documented in 95 (85%) cases, 59 (62%) of which (62%) were localized in the lower extremities. Final outcome dichotomized either as death patient (group one) or surviving patient (group two) was documented in 103 cases of whom 67 (65%) were in group one and 36 (35%) were in group two. By comparing them, differences were seen in mean age (P = 0.004, 95% CI 3.9 to 19.8), age categories (< 50 vs. > or = 50) (chi 2 P = 0.001, 95% CI 0.6 to 0.60), severity of tetanus (Fisher exact test P = 0.0009, 95% CI 2 to 53) and mean hospitalization time (mean difference 14.8, P = 0.0001, 95% CI 11 to 18) but not in sex (chi 2 0.69, P = 0.40). CONCLUSIONS: In the State of Yucatan, Peninsula de Yucatan, Mexico, TIAP is still an endemic process with high mortality rate specially among young productive people. Secondary preventive measures as routinely tetanus toxoid booster vaccination are still not enough, at least in adulthood.

Effect of solvent on the preparation of surfactant-free poly(DL-lactide-co-glycolide) nanoparticles and norfloxacin release characteristics.

The surfactant-free nanoparticles of poly(DL-lactide-co-glycolide) (PLGA) were prepared by dialysis method without surfactant and physicochemical properties such as particle size and drug contents were investigated against used initial solvent. The size of PLGA nanoparticles and drug contents were significantly changed by used initial solvent. The size of PLGA nanoparticles prepared from dimethylacetamide (DMAc), dimethylformamide (DMF), and dimethylsulfoxide (DMSO) as a initial used solvent was smaller than that of acetone. Selected initial solvent used to dissolve the copolymer significantly affects the size of nanoparticles and drug contents. It was shown that PLGA nanoparticles have spherical shapes from the results of scanning electron microscopy (SEM) and transmission electron microscopy (TEM) observations. It was thought that surfactant-free nanoparticles of PLGA entrapping norfloxacin (NFX) has nice drug loading capacity without free-drug on the surface of nanoparticles through the analysis of X-ray powder diffraction. From these results, it was showed the potential that the PLGA nanoparticles could be formed successively by dialysis method without surfactant. Release kinetics of NFX used as a model drug was governed by not only drug contents but also particle size parameter. The higher the drug contents and the larger the particle size resulted in slower the drug release.

Effect of ginsenosides, active components of ginseng, on capsaicin-induced pain-related behavior.

Our recent study demonstrated that ginsenosides had antinociceptive effects by reducing some types of pain-related behavior in mice (Yoon et al., 1998. Ginsenosides induce differential antinociception and inhibit substance P-induced nociceptive response in mice. Life Science 62, PL319-PL325). In the present study we further investigated whether ginsenosides produce antinociceptive effects through an action at central or peripheral site(s) and whether these effects are mediated by the opioid system. Intraperitoneally injected ginsenosides suppressed in a dose-dependent manner the pain-related behavior produced by capsaicin injection into the plantar surface of the hind paw; the ED(50) was 49 mg/kg [26-92 mg/kg, 95% confidence interval (C.I.)]. Intrathecally or intracerebroventricularly administered ginsenosides also suppressed the capsaicin-induced pain-related behavior in a dose-dependent manner; the ED(50)s were 1.72 mg/kg (0.8-3.72 mg/kg, 95% C.I.) and 1. 48 mg/kg (0.8-2.6 mg/kg, 95% C.I.), respectively. On the other hand, subcutaneously injected ginsenosides to the plantar surface prior to the capsaicin injection did not alter the pain-related behavior. Naloxone pretreatment was without effect in blocking the antinociceptive effect of intrathecally administered ginsenosides. Intraperitoneally injected ginsenosides also did not significantly affect the motor response of animals. These results suggest that ginsenosides produce antinociceptive effects through their action at the spinal and/or supraspinal site(s), not at nociceptors in the periphery. In addition, the results suggest that the antinociceptive effects are not mediated by opioid receptors.

In vivo antioxidant effect of green tea.

OBJECTIVE: The object of this study was to investigate the in vivo antioxidant effect of green tea and dosage effect of green tea on antioxidant effect. DESIGN: We tested 10 healthy subjects (aged 23-25 y, five women and five men) with overnight fasting. The total antioxidant capacity of plasma was measured at baseline and 60 min and 120 min after ingestion of 150 ml green tea. Green tea was prepared by infusing 2.5 g of dried green tea leaves for 2 min at 80 degrees C in 150 ml of water. In the second week, they took 300 ml of tea (5.0 g of green tea leaves) and, in the third week, 450 ml of tea (7.5 g of green tea leaves). The total antioxidant capacities of plasma were determined with a Total Antioxidant Kit (Randox Laboratories Ltd, UK) using a Cobas Mira analyser (Roche Diagnostic Systems Inc., Switzerland). The mean intra-assay coefficient of variation was 1.2%. RESULTS: The total antioxidant capacity of plasma increased by 1.1% at 60 min and 2.1% at 120 min over baseline value in subjects consuming 150 ml of green tea, which was statistically not significant. However, total antioxidant capacity of plasma after consuming 300 ml of green tea showed a significant increase of 7.0% after 60 min and 6.2% after 120 min (P<0.0001), and after consuming 450 ml 12.0% after 60 min and 12.7% after 120 min over baseline value (P<0.0001). CONCLUSIONS: Total antioxidant capacity of plasma was significantly increased after taking green tea in amounts of 300 and 450 ml. A positive increment according to green tea dosage was also observed. SPONSORSHIP: This work was funded by the Pacific Corporation (Korea).

Cloning of a sesquiterpene cyclase and its functional expression by domain swapping strategy.

Sesquiterpene cyclase, the first committed step enzyme from the general isoprenoid building block farnesyl pyrophosphate (FPP) for the synthesis of phytoalexin capsidiol, was isolated from the UV-C treated leaves of Capsicum annuum. This sesquiterpene cyclase, termed as CASC2 showing 77% amino acid identity with the previously cloned sesquiterpene cyclase CASC1, was composed of 560 amino acids with a calculated molecular mass of 64,907. The mRNA expression pattern of CASC2 was very similar to that of CASC1 during the time course of UV-C irradiated leaves of pepper on RNA blot analysis by using each specific probe. The heterologous expression in Escherichia coli using the CASC2 full length failed; however the chimeric construct of CASC2 in which the amino terminal 164 amino acid substituted by the equivalent portion of either CASC1 or tobacco sesquiterpene cyclase was capable of expressing the functional sesquiterpene cyclase activities. The radio-labeled enzymatic products catalyzed by the partially purified chimeric CASC2 were comigrated with authentic radio-labeled sesquiterpene on thin layer chromatography.

Clonazepam release from core-shell type nanoparticles of poly(epsilon-caprolactone)/poly(ethylene glycol)/poly(epsilon-caprolactone) triblock copolymers.

The triblock copolymer based on poly(epsilon-caprolactone) (PCL) as hydrophobic part and poly(ethylene glycol) (PEG) as hydrophilic one was synthesized and characterized. Core-shell type nanoparticles of poly(epsilon-caprolactone)/poly(ethylene glycol)/poly(epsilon-caprolactone) (CEC) block copolymer were prepared by a dialysis technique. According to the amphiphilic characters, CEC block copolymer can self-associate at certain concentration and their critical association concentration (CAC) was determined by fluorescence probe technique. CAC value of the CEC-2 block copolymer was evaluated as 0.0030 g/l. CAC values of CEC block copolymer decreased with the increase of PCL chain length, i.e. the shorter the PCL chain length, the higher the CAC values. From the observation of transmission electron microscopy (TEM), the morphologies of CEC-2 core-shell type nanoparticles were spherical shapes. Particle size of CEC-2 nanoparticles was 32.3+/-17.3 nm as a monomodal and narrow distribution. Particle size, drug loading, and drug release rate of CEC-2 nanoparticles were changed by the initial solvents and the molecular weight of CEC. The degradation behavior of CEC-2 nanoparticles was observed by 1H NMR spectroscopy. It was suggested that clonazepam (CNZ) release kinetics were dominantly governed by diffusion mechanism.

Ginsenosides inhibit capsaicin-activated channel in rat sensory neurons.

Ginsenosides isolated from ginseng are biologically active components. In this study, whole-cell and inside-out configurations of patch clamp technique had been used to test the effect of ginsenosides on the capsaicin-activated channels in cultured small diameter sensory neurons of young rat. Ginsenosides (100 microg/ml) decreased the amplitude of capsaicin-activated currents by 78.2% in whole cell mode. Similarly, ginsenosides decreased capsaicin-activated single-channel activities in a dose-dependent manner in inside-out patches. These results indicate that ginsenosides might directly block capsaicin-activated channels, resulting in attenuation of the currents in rat sensory neurons.

Preparation of monoclonal antibody against ginsenoside Rf and its enzyme immunoassay.

A rapid and sensitive indirect competitive enzyme immunoassay method has been developed for quantitating ginsenoside Rf (Rf) in crude total Panax ginseng saponins and in rat plasma using high titer mouse monoclonal antibody (mAb) raised against a conjugate of Rf and bovine serum albumin (BSA). The isotype of mAb against Rf was IgG3 with a K chain. The presence of Rf inhibited the binding of the mouse anti-Rf mAb to a Rf-BSA solid phase coating antigen. The working range was 0.01-10 ng/assay and detection limits were 20 pg in various ginseng extract fractions or 34 pg in rat plasma per assay. The anti-Rf mAb cross-reacted with ginsenoside Rg2 by 57.5%, but not with other ginsenosides. However, this anti-Rf mAb did not cross-react with BSA or cellubiose, which is a carbohydrate component of Rf. Using this standard curve, we could measure the amount of Rf in ginseng total extract, ginseng total saponins, protopanaxadiol saponins, and propanaxatriol saponins. We could also measure the amount of Rf in rat plasma after the oral administration of Rf and found that Rf reached a maximum level in rat plasma after 16 h. These results indicate that the anti-Rf mAb could be useful for the quantitation of Rf in crude ginseng fractions and in body fluids.

[Acute pesticide poisoning]. / Intoxicación aguda por plaguicidas.

OBJECTIVE: To describe the epidemiologic pattern of acute pesticide poisoning (APP) in a general hospital in Merida, Yucatan, Mexico. MATERIAL AND METHODS: From 1994 to 1998, 33 patients 13 years of age or older with diagnosis of APP were studied. Descriptive statistics were used to analyze information. RESULTS: Males were frequently affected (82%), specially those coming from rural areas (60%). The mean age of the group was 34 +/- 15.8 years. In 79% of the cases, pesticides were used to commit suicide and 33% of poisoning cases were due to organophospate pesticides. The mortality rate was 12%. CONCLUSIONS: In this small sample, acute poisoning from pesticides in the agricultural setting may be underestimated, since it was less frequent than in the general population. APP was more commonly used by indigent people to commit suicide.

Characterization and expression of a novel alternatively spliced human angiopoietin-2.

Angiopoietin-2 (Ang2) is a naturally occurring antagonist of angiopoietin-1 (Ang1) that competes for binding to the Tie2 receptor and blocks Ang1-induced Tie2 autophosphorylation during vasculogenesis. Using the polymerase chain reaction, we isolated a cDNA encoding a novel shorter form of Ang2 from human umbilical vein endothelial cell cDNA and have designated it angiopoietin-2(443) (Ang2(443)), because it contains 443 amino acids. Part of the coiled-coil domain (amino acids 96-148) is absent in Ang2(443) because of alternative splicing of the gene. Like Ang2, recombinant Ang2(443) expressed in COS-7 cells is secreted as a glycosylated homodimeric protein. Recombinant Ang2(443) binds to the Tie2 receptor but does not induce Tie2 phosphorylation. Pre-occupation of Ang2(443) on Tie2 inhibits Ang1 or Ang2 binding and inhibits Ang1-induced phosphorylation. Expression of Ang2(443) mRNA is detectable in primary endothelial cells, several nonendothelial tumor cell lines, and primary tumor tissues. Interestingly, two cervical carcinoma cell lines express relatively moderate levels of Ang2(443) mRNA and protein. Macrophages express mainly Ang2 mRNA, but the expression of Ang2(443) mRNA is temporarily up-regulated during macrophage differentiation. These results suggest that Ang2(443) is a functional antagonist of Ang1 and could be an important regulator of angiogenesis during some tumorigenic and inflammatory processes.

Distribution of haptoglobin phenotypes in a Korean population, using the semi-automated PhastSystem.

We have established a new phenotyping method for haptoglobin, based on sodium dodecyl sulphate-polyacrylamide gel electrophoresis using the PhastSystem (Pharmacia Biotech, Uppsala, Sweden), followed by immunoblotting for detection. We measured haptoglobin concentrations and determined the haptoglobin phenotypes of 316 healthy Koreans using this method: 31 (9.8%) were of Hp 1-1 type, 140 (44.3%) of Hp 2-1 type and 145 (45.9%) of Hp 2-2 type. The haptoglobin allele frequencies were calculated to be 0.32 for Hp1 and 0.68 for Hp2. We were able to visualize up to 12 bands from the human Hp 2-2 polymeric series, with molecular weights in the range 171.9 x 10(3) to 802.2 x 10(3). The reference range of serum haptoglobin concentrations obtained by the IFCC (International Federation of Clinical Chemistry) standard method was 0.27-2.14 g/L. The serum haptoglobin concentration in Koreans was similar to that of Caucasians, but the Hp1 allele frequency was lower in Koreans. Our method could be used in clinical laboratories as a simple and practical method of haptoglobin phenotyping. In addition, the Hp 2-2 polymeric series could be used as high molecular weight standards.