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PubMed2005

Selenium extractability from a contaminated fine soil fraction: implication on soil cleanup.

Two batches of fine soil fraction of an acidic soil were deliberately contaminated with selenite (Se(IV)) and selenate (Se(VI)), respectively, and aged for more than 220 days. Speciation analysis using continuous flow-through hydride generation atomic absorption spectrometry (HGAAS) indicated that the species were predominant in their respective aged soils. A selective sequential extraction scheme was employed to fractionate the Se retained in the soils into six fractions of varying retentions. Abilities of various chemical reagents in extracting the Se in the two soil batches were then evaluated. The reagents investigated were sodium salts such as sodium chloride (NaCl), sodium sulfate (Na2SO4), sodium carbonate (Na2CO3), and sodium phosphate (Na3PO4), and two oxidants, namely, hydrogen peroxide (H2O2) and potassium permanganate (KMnO4). It was found that NaCl, Na2SO4, and Na2CO3 could only extract the exchangeable fraction of Se, while Na3PO4 could extract the exchangeable and strongly-bound fractions. Selenate was extracted more than Se(IV) by the salts. The kinetics of Se(IV) extraction by Na3PO4 could be best described by the Elovich model, while the Ritchie second-order model was the most appropriate to describe Se(VI) extraction. Efficiencies of the oxidants in Se(IV) extraction highly depended on their applied dosages. Both H2O2 and KMnO4 were able to extract greater than 93% of total Se, and therefore were significantly more effective than the salts in Se(IV) extraction.

Chemical Fractionation↗

PubMed2000

Applications of video mixing and digital overlay to neuroethology.

Neuroethological experiments often require video images of animal behavior and recordings of physiological data to be acquired simultaneously, synchronized with each other, stored, and analyzed together. The use of inexpensive multimedia computers offers new possibilities for mixing video images, analog voltages, and computer data, storing these combined signals to videotape, and extracting quantitative data for analysis. In this paper, we summarize methods for mixing images from multiple video cameras and a Macintosh computer display to facilitate manipulation of data generated during our neurophysiological and behavioral research. These technologies enhance accuracy, speed, and flexibility during experiments, and facilitate selecting and extracting quantitative data from the videotape for further analysis. Three applications are presented: (A) we used an analog video mixer to synchronize neurophysiological recordings with ongoing behaviors of freely moving rats; (B) we used a chroma keyed digital overlay to generate positional data for the rat's face during drinking behavior; and (C) we combined a computer model of a rat's head and whiskers with videos of exploratory behaviors to better track and quantify movements in three dimensions. Although the applications described here are specific to our neuroethological work, these methods will be useful to anyone wishing to combine the signals from multiple video sources into a single image or to extract series of positional or movement data from video frames without frame grabbing.

Animals↗

PubMed1989

Monoclonal antibodies reactive with components in serum-free conditioned medium from a human breast cancer cell line (MCF-7).

Approximately 10,000 primary hybridomas were generated after immunization with either serum-free conditioned medium (SFCM) or extracts from the human breast cancer cell line MCF-7. A total of 11 different monoclonal antibodies (MAbs; 8 generated against SFCM and 3 generated against cell extracts) were selected on the basis of high specificity in cell-binding ELISA for human breast cancer cell lines. The 8 different MAbs obtained by immunization with SFCM all reacted with secreted components in SFCM from MCF-7 cells and 4 of these MAbs reacted with glycolipids extracted from MCF-7 cells. 1 of these MAbs (S2) also recognized a secreted glycoprotein of approximately 77 kilodaltons (kDa). The remaining 4 MAbs did not show specificity solely for carbohydrate determinants. 1 of these MAbs (S7) recognized a secreted protein of approximately 41 kDa. The 3 MAbs raised against cell extracts from breast cancer cells reacted with cytoplasmic antigens in immunofluorescence but also reacted with a secreted component in SFCM from MCF-7 cells. Immunoblotting experiments with proteins from cell extracts and with proteins in SFCM showed that these antibodies all reacted with a protein of a molecular weight of approximately 40 kDa. Our results suggest that this component is cytokeratin 19 or proteolytically processed cytokeratin 19.

Antibodies, Monoclonal↗

PubMed1994

Item-level and scale-level factor structures of the MMPI-A.

The Minnesota Multiphasic Personality Inventory-Adolescent (MMPI-A) is a 478-item test that represents a substantial revision of the original form of the MMPI. This investigation sought to identify the item-level factor structure of the MMPI-A and also examined the scale-level factor structure of the 69 scales and subscales of this instrument. The study utilized the 1,620 normal adolescents (805 boys and 815 girls) of the normative sample for the MMPI-A. These adolescents ranged in age from 14 to 18, inclusive, with a mean age of 15.54 for boys and 15.60 for girls. A principal factor analysis of item-level responses resulted in extraction of 14 factors that were subjected to promax (oblique) rotation procedures. These 14 factors incorporated 81% of the total MMPI-A item pool and accounted for 44% of the total item-level response variance. For the scale-level analysis, 8 factors were selected for extraction and submitted to promax rotation procedures. These eight factors accounted for a total of 93.5% of the total variance in MMPI-A scale and subscale raw scores. Item-level results were discussed in terms of areas of congruence and dissimilarities from previously reported MMPI factor analyses in adolescent and adult samples, and scale-level factor results were presented in terms of clinical implications for the interpretation of MMPI-A scales and subscales.

Adaptation, Psychological↗

PubMed2000

Is pseudophakic astigmatism a desirable goal?

PURPOSE: To determine whether pseudophakic astigmatism is a desirable goal, and if so, which one is better: against-the-rule (ATR) or with-the-rule (WTR). METHOD: Eyes were included only if they had an uncorrected vision > or = 6/18 and N/18. Three groups, of 40 patients each were evaluated: group 1, pseudophakes with neutral astigmatism; group 2, with ATR and group 3, with WTR astigmatism Unaided distance and near visual acuity was recorded. Statistical analysis was performed using the chi-square test for independence. RESULTS: Unaided distance vision of > or = 6/7.5 was achieved in 19 eyes (47.5%) of group 1 (neutral), 12 eyes (30%) in group 2 (ATR), and 5 eyes (12.5%) in group 3 (WTR) (p = 0.0133, significant). Unaided near vision of > or = N/9 was achieved in 17 eyes (42.5%) in group 1 (neutral), 34 eyes (85%) in group 2 (ATR), and 10 eyes (25%) in group 3 (WTR) (P < 0.001, significant). Group 1 (neutral) fared the best for unaided distance visual acuity. Group 2 (ATR) was better than in group 3 (WTR) for distant vision. Group 2 (ATR) fared the best for unaided near vision. CONCLUSION: ATR astigmatism could be a desirable goal after cataract extraction in selected populations because the largest proportion of these cases achieved good unaided near vision with acceptable distant vision.

Astigmatism↗

PubMed2005

Transcription factors as targets of the anti-inflammatory treatment. A cell culture study with extracts from some Mediterranean diet plants.

During the inflammatory response at least 2 transcription factors, NF-kappaB and AP-1, are involved in the altered profile of gene expression. We used human hepatoma (HepG2) and human umbilical vein endothelial cells (HUVEC) as a model system: NF-kappaB and AP-1 were activated by the proinflammatory cytokine IL-1 in the absence or presence of 21 selected plant extracts and the effect was evaluated by the electrophoretic mobility shift assay (EMSA). In both types of cells activation of NF-kappaB by IL-1 was significantly inhibited by extracts from Scandix australis and Artemisia alba, whereas extracts from Amaranthus sp., Eryngium campestre, Thymus pulegioides and Reichardia picroides elicited cell-type dependent response. The IL-1-induced AP-1 activation was diminished by extracts from Scandix australis, Amaranthus sp. and Artemisia alba more potently in HUVEC, while extracts from Urospermum picroides and Scandix pecten-veneris in HepG2 cells. Inhibitory activities of plant extracts towards cytokine activated NF-kappaB and AP-1 depend to some extent on the order of addition of IL-1 and plant extract to the cell culture, but the mechanism of action of extract components is not clear: although plant polyphenols may participate they are unlikely to be the only mediators, and MAP kinases were found generally not involved in down-regulation of transcription factors activity by plant extracts.

Acetylcysteine↗

PubMed1998

A review of journal clubs in postgraduate medical education.

OBJECTIVE: To review the goals, organization, and teaching methods of journal clubs, summarize elements of successful clubs, and evaluate their effect on reading habits, and effectiveness in meeting teaching goals. Examples of clubs that utilize principles of adult learning are reviewed. DATA SOURCES: English language articles identified through a MEDLINE search (1966-1997) using the MeSH terms "internship" and "residency," and text words "journal club" and "critical appraisal." STUDY SELECTION: Articles on learning goals and organization were included if they represented national or regional surveys with a response rate of 65% or greater. Articles that evaluated teaching effectiveness were included if they used a controlled, educational design, or if they exemplified important adult learning principles. DATA EXTRACTION: Data were manually extracted from selected studies and reviews. DATA SYNTHESIS: A major goal for most clubs is to teach critical appraisal skills. Clubs with high attendance and longevity are characterized by mandatory attendance, availability of food, and perceived importance by the program director. Residents who are taught critical appraisal report paying more attention to the methods and are more skeptical of the conclusions, and have increased knowledge of clinical epidemiology and biostatistics, but studies have failed to demonstrate that these residents read more, or read more critically. Reading guidelines may be useful for teaching critical appraisal skills, and may be associated with increased resident satisfaction. CONCLUSIONS: Journal club formats are educationally diverse, can incorporate adult learning principles, and are an adaptable format for teaching the "new basic sciences."

Adult↗

PubMedDate not supplied

Comparison of preoperative ibuprofen, acetaminophen, and placebo administration on the parental report of postextraction pain in children.

Previous investigations demonstrated that administering a preoperative analgesic can reduce postextraction pain in adults. Studies also have shown that ibuprofen was superior in alleviating postextraction pain when compared with acetaminophen. The purpose of this study was to compare the efficacy of the preoperative administration of ibuprofen, acetaminophen, and placebo in reducing postextraction pain in children. Sixty children, aged 2-10 years, requiring primary teeth extractions, were selected to participate in the study. Patients were assigned in a blind, random fashion to one of the three pretreatment drug groups. Parental report of their child's pain and use of analgesics during the immediate 7-hr postoperative period was elicited by telephone the day after the extraction procedure. Thirty-five percent of the patients were reported by their parents to experience postextraction pain. Of those patients, 52% received postoperative analgesics for pain relief (18% of the total). Although there was a trend toward reduced postextraction pain reported by the parents, the preoperative administration of neither analgesic was found to be statistically superior by chi-square analysis to placebo administration.

Acetaminophen↗

PubMed1995

Biochemical, antigenic and allergenic characterization of crude extracts of Drechslera (Helminthosporium) monoceras.

In a previous study with airborne mould extracts we verified that Drechslera (Helminthosporium) monoceras presented stronger reactions than those presented by 42 other moulds isolated in São Paulo city. In the present study, we evaluated the biochemical composition and the antigenicity of crude extracts obtained from vegetative and conidial stage of D. monoceras using Czapeck broth (CB) modified and tris-HCl for extraction. The maximum values of total proteins and lipids were verified in the crude extract obtained in the 28th day of growth, and maximum values of carbohydrates were observed in the extracts of the 16th, 22nd and 26th days. The fractionated proteins by SDS-PAGE presented bands with molecular weights between 14.4 to 67 Kd; the 28th day extract showed a larger number of bands. The carbohydrates and amino acids were characterized by thin-layer chromatography. The antigenicity of the crude extracts was verified by immunodiffusion reaction in agar against rabbit hyperimmune sera. Precipitation lines were observed in all studied extracts and common antigenic molecular populations. Based on the above results, the 28th day extract was selected to verify the induction of IgE antibody responses in immunizations of Balb/c and cAF-1 mice, and titer by passive cutaneous anaphylaxis test using Wistar rats. The maximum titers obtained were 160 in cAF-1 mice and 1.280 in Balb/c mice. The results suggest that the 28th day extract contains allergenic fractions and should be chosen for future studies related to fractionation, characterization and standardization in diagnostic methods and immunotherapy.

Allergens↗

PubMed2006

Expression of avian reovirus sigmaC protein in transgenic plants.

Avian reovirus (ARV) structural protein, sigmaC encoded by S1 genome segment, is the prime candidate to become a vaccine against ARV infection. Two plant nuclear expression vectors with expression of sigmaC-encoding gene driven by CaMV 35S promoter and rice actin promoter were constructed, respectively. Agrobacterium containing the S1 expression constructs were used to transform alfalfa, and transformants were selected using hygromysin. The integration of S1 transgene in alfalfa chromosome was confirmed by PCR and histochemical GUS staining. Western blot analysis using antiserum against sigmaC was carried out to determine the expression of sigmaC protein in transgenic alfalfa cells. The highest expression levels of sigmaC protein in the cellular extracts of selected p35S-S1 and pAct1-S1 transgenic alfalfa lines were 0.008% and 0.007% of the total soluble protein, respectively. The transgenic alfalfa cells with expression of sigmaC protein pave the way for the development of edible vaccine.

Capsid Proteins↗

PubMed2003

Development and evaluation of an isotope dilution LC/MS method for the determination of total homocysteine in human plasma.

Elevated plasma homocysteine has been identified as a strong and independent risk factor for cardiovascular diseases, and recently, it has been associated with the development of dementia in older adults. Selected ion-monitoring isotope-dilution LC/MS (electrospray) has been developed and evaluated as a reference method for the accurate determination of total homocysteine in human plasma. Homocysteine is quantitatively isolated from plasma via the use of anion-exchange resins and then detected and quantified in stabilized plasma extracts with selected ion-monitoring LC/MS. This method is shown to be highly comparable to LC/MS/MS determinations in terms of its analytical accuracy and precision, yet this alternative measurement approach does not necessitate the enhanced instrumentation or added expense required of tandem MS/MS determinations. LC/MS detection of homocysteine was linear (standard error of the estimate for the regression line was 0.0323) over 3 orders of magnitude, and the calculated limits of detection and quantification were 0.06 micromol/L (0.12 ng on column) and 0.6 micromol/L (1.2 ng on column), respectively. Independent calibration curves showed excellent linearity (r2 > or = 0.996) between 0 and 25 micromol/L homocysteine over a 3-day period. The accuracy and precision of total homocysteine measurements for patient samples and quality control pools using LC/MS were compared to total homocysteine measurements using LC/MS/MS, GC/MS, FPIA, and LC-FD. LC/MS performed well in relation to the other homocysteine methods in terms of its capability to accurately quantify plasma homocysteine over the normal range (5-15 micromol/L).

Blood Chemical Analysis↗

PubMed2006

Accurate mass-driven analysis for the characterization of protein phosphorylation. Study of the human Chk2 protein kinase.

We describe the data-dependent analysis of protein phosphorylation using rapid-acquisition nano-LC-linear quadrupole ion trap Fourier transform ion cyclotron resonance mass spectrometry (nano-LC-FTMS). The accurate m/z values of singly, doubly, and triply charged species calculated from the theoretical protonated masses of peptides phosphorylated at all Ser, Thr, or Tyr residues of the human checkpoint 2 (Chk2) protein kinase were used for selected ion extraction and chromatographic analysis. Using a kinase-inactive Chk2 mutant as a control, accurate mass measurements from FTMS and collision-induced dissociation spectra, 11 novel Chk2 autophosphorylation sites were assigned. Additionally, the presence of additional Chk2 phosphorylation sites in two unique peptides was deduced from accurate mass measurements. Selected ion chromatograms of all Chk2 phosphopeptides gave single peaks except in three cases in which two closely eluting species were observed. These pairs of phosphopeptides were determined to be positional isomers from MS/MS analysis. In this study, it was also found that ions due to the neutral loss of phosphoric acid from the parent peptide ion were not prominent in 18 of 36 MS/MS spectra of O-linked Chk2 phosphopeptides. Thus, accurate mass-driven analysis and rapid parallel MS/MS acquisition is a useful method for the discovery of new phosphorylation sites that is independent of the signature losses from phosphorylated amino acid residues.

Amino Acid Sequence↗

PubMed2005

RP-HPLC analysis of the phenolic compounds of plant extracts. investigation of their antioxidant capacity and antimicrobial activity.

Extracts of aromatic plants of Greek origin were examined as potential sources of phenolic compounds. RP-HPLC with UV detection was employed for the identification and quantification of the phenolic antioxidants, present in methanolic extracts. The most abundant phenolic acids were ferulic acid (1.1-280 mg/100 g of dry sample) and caffeic acid (1.2-60 mg/100 g of dry sample). (+)-Catechin and quercetin were the most abundant flavonoids. Apigenin and luteolin were detected in high amounts in Menta pulegium and Thymus vulgaris, respectively. The antioxidant capacity was determined, in dried ground plants and in their methanol extracts, with the Rancimat test using sunflower oil as substrate. Both pulverized plants and extracts showed antioxidant capacity. Total phenolic content in the extracts was determined spectrometrically according to the Folin-Ciocalteu assay and ranged from 1 to 21 mg of gallic acid/100 g of dry sample. Antimicrobial activity of the extracts against selected microbes was also conducted in this study.

Anti-Bacterial Agents↗

PubMed2002

Chemical sequential extraction for metal partitioning in environmental solid samples.

A comprehensive review of the sequential extraction schemes for metal fractionation in environmental samples (ie., sediment, soil, sewage sludge, fly ash, etc.) is presented. The review contains more than 400 references and covers principally the literature published over the last decade. The use of each reagent involved in these schemes is looked at critically, and guidelines for their selectivity and extraction capacity are given. The operational character of these schemes is emphasised. Topics such as comparability between sequential extraction schemes of widespread use, harmonisation, acceleration, validation, etc. are addressed and future developments outlined.

Chemistry Techniques, Analytical↗

PubMed2006

The influence of haemostatic agents on healing of healthy human dental pulp tissue capped with calcium hydroxide.

AIM: To investigate the hypothesis that different haemostatic agents could impair the histological response of human pulps capped with calcium hydroxide. METHODOLOGY: Forty-five third molars scheduled for extraction were selected. Class I cavities with pulp exposures were prepared. Three agents were used to control bleeding: 0.9% saline solution (control, n = 14), 5.25% sodium hypochlorite (n = 16) and 2% chlorhexidine digluconate (n = 15). The pulps were dressed with hard-setting calcium hydroxide cement. After 7, 30 or 90 days, teeth were extracted, formalin-fixed and prepared for histochemical techniques. The biological response was categorized using the following criteria: inflammatory response, soft tissue organization, reactionary dentine and reparative dentine. Data were submitted to statistical analysis, using nonparametric Kruskal-Wallis one-way analysis of variance on ranks. Differences amongst groups were detected using Dunn's method. RESULTS: The statistical analysis disclosed that whilst inflammatory response decreased over time, reactionary dentine deposition and reparative dentine formation increased in the latter periods of evaluation (P < 0.05). The three agents had similar performances for all criteria evaluated. The conventional pulp response to calcium hydroxide was observed over time, and complete pulp healing was observed in 88% of the specimens after 90 days. CONCLUSION: The three haemostatic agents did not impair the healing process following pulp exposure and capping with calcium hydroxide at different time intervals investigated.

Adult↗

PubMed1993

A plasmid system to monitor gene conversion and reciprocal recombination in vitro.

A plasmid system allowing for the detection of recombinagenic activities in cell-free extracts is described. Two truncated alleles of the bacterial neomycin resistance gene (neo), differing from each other at a polymorphic restriction site, were constructed. Recombinations involving both alleles mediated by Drosophila embryo nuclear protein extracts or Drosophila larva whole cell protein extracts were selected by their ability to confer kanamycin resistance to E. coli. Restriction analysis of plasmids recovered from E. coli transformants allowed the monitoring of the two molecular mechanisms which can lead to functional neo genes, gene conversion and reciprocal recombination. A dose dependent increase in the recombination frequency with increasing amounts of cell extract was observed. Recombination was further increased by linearizing one of the two substrate plasmids. The Drosophila cell extracts catalyzed recombination in vitro since after incubation a recombination product could be identified by polymerase chain reaction (PCR) technology. The recombination was absolutely dependent on the presence of an active cell extract, since no diagnostic PCR product was detected in a reaction where extract was omitted. Analysis of a representative number of recombinant plasmids by restriction analysis revealed that in the absence of an exogenous recombinational system less than 2% of kanamycin resistant recombinant plasmids occurred by gene conversion upon transformation into E. coli. In contrast, recombinants exhibiting restriction patterns diagnostic for gene conversion were observed at frequencies between 5.1% and 9.8% after incubation with Drosophila larva cell extracts. These results strongly argued that gene conversion is a prominent mechanism of recombination in Drosophila mitotic cells.

Animals↗

PubMed2003

Sperm immobilization activity of Allium sativum L. and other plant extracts.

AIM: To identify possible spermicidal agents through screening a number of edible medicinal plants with antimicrobial activity. METHODS: Initial screening was made on the basis of ram cauda epididymal sperm immobilization immediately after addition of extracts. The most potent extract was selected and was evaluated on both ram and human spermatozoa. To unravel its mode of action several sperm functional tests were carried out, namely viability of cells, hypo-osmotic swelling test for membrane integrity and assays of membrane-bound enzyme 5'-nucleotidase and acrosomal marker enzyme acrosin. RESULTS: The crude aqueous extract of the bulb of Allium sativum L. showed the most promising results by instant immobilization of the ram epididymal sperm at 0.25 g/mL and human ejaculated sperm at 0.5 g/mL. Sperm immobilizing effects were irreversible and the factor of the extract responsible for immobilization was thermostable up to 90 deg. On boiling at 100 deg for 10 minutes, this activity was markedly reduced. Moreover, this extract was able to cause aggregation of ram sperms into small clusters after 30 minutes of incubation at 37 deg. However this property was not found in human spermatozoa. More than 50 % reduction in sperm viability and hypo-osmotic swelling occurred in treated sperm as compared with the controls, indicating the possibility of plasma membrane disintegration which was further supported by the significant reduction in the activity of membrane bound 5'-nucleotidase and acrosomal acrosin. CONCLUSION: The crude aqueous extract of A. sativum bulb possesses spermicidal activity in vitro.

5'-Nucleotidase↗

PubMed1980

Isoenzymes of creatine kinase in extracts of various parts and regions of the human central nervous system.

The distribution of isoenzymes of creatine kinase (EC 2.7.3.2.) was investigated by agarose gel electrophoresis of extracts of selected parts and regions of the human central nervous system. Besides the major brain isoenzyme BB, we demonstrated the presence of three other creatine kinase isoenzyme forms. The distribution of creatine kinase isoenzymes depending strongly on the region from which the biopsy was taken. We found substantial amounts of the MB isoenzyme in extracts of the dura from the cauda equina of the two adults examined.

Adult↗