bluuebunny/crossref_metadata_2025 huggingface dataset:
Successful use of two rapid HCV assays in a high prevalence romanian population
Authors: Constantine Niel T. , Holm‐Hansen Carol , Skaug Nils , Vasilescu Florica
Published: March 2007

Hepatitis C virus (HCV) infection is of significant concern for recipients of blood and for patients who share resources such as hemodialysis machines. In many developing countries and in small peripheral laboratories, facilities and capabilities to perform ELISA assays may not be available. We evaluated two newly marketed rapid HCV serologic assays to determine their ability and suitability to detect antibodies to HCV in hemodialysis patients in Bucharest, Romania. Results indicated that both the Rapid HCV Ab assay (Clonatec, Paris, France) and the HCV‐SPOT (Diagnostic Biotechnology, Singapore) detected HCV antibody in 23 of 27 patients. All 23 samples were also reactive by a routine HCV ELISA and were confirmed using a supplemental assay (HCV blot, Diagnostic Biotechnology, Singapore). Only one sample produced equivocal result by the Rapid HCV Ab assay. In this high prevalence population (90%), the use of recombinant or synthetic peptide rapid HCV assays has been successful in detecting confirmed positive cases of HCV and has shown excellent correlation with an ELISA screening assay. The tests are simple to perform, can be performed by individuals with minimal training, and have built‐in quality control measures. We conclude that these tests may have important applicability in certain testing situations. © 1994 Wiley‐Liss, Inc.
Current status of available standards for quality improvement of assays for detection of autoantibodies to nuclear and intracellular antigens
Authors: Nakamura Robert M. , Bylund David J. , Tan Eng M.
Published: March 2007

Considerable progress has been made in the improvement of clinical assays for the detection of autoantibodies to nuclear and intracellular antigens with the use of available World Health Organization (WHO) and Arthritis Foundation/Centers for Disease Control (AF/CDC) standards. The ultimate goal of standardization is for various clinical laboratory test results to be interchangeable and for an exchange of data to be done with confidence.

This report discusses the available standards. In addition, significant technical problems and variations in methodologies for the detection of autoantibodies to intracellular antigens noted during a 4‐year study by a European Consensus Study Group are detailed.

Currently, there is a need for a future generation of reference preparations and standards that will show specific antibody reactivity on sensitive enzymes and immunoblotting assays. Standardization efforts should be done to characterize specific nuclear and cellular antigen preparations that may be of natural or of recombinant technology origin. © 1994 Wiley‐Liss, Inc.
Chemiluminescent immunoassays: Discrimination between the reactivities of natural and human patient antibodies with antigens from eukaryotic pathogens, Trypanosoma cruzi and Paracoccidioides brasiliensis
Authors: Almeida Igor C. , Rodrigues Elaine G. , Travassos Luiz R.
Published: March 2007

Quantitative chemiluminescent enzymelinked immunosorbent assay (ELISA) and dot‐blotting procedures were developed to evaluate the reactivity of human antibodies with crude antigens and purified molecules of parasites and fungi, mainly Trypanosoma cruzi and Paracoccidioides brasiliensis. Reproducible, highly sensitive, and strictly doseresponding results were obtained, with the specificity depending on the kind of antigen used. Mixed antigens (epimastigote membrane and HIV‐1 heptapeptide) applied in dots could be independently recognized by specific sera. Purified antigens (T. cruzi F2/3 and P. brasiliensis gp43) at very small concentrations gave specific reactions with patients' sera diluted ≥1:1,000 and were very poorly reactive or unreactive with natural antibodies using the chemiluminescent immunoassays. P. brasiliensis crude antigen Fava Netto polysaccharide antigen (FNPA) contained peptide epitopes recognized by natural antibodies and carbohydrate epitopes reactive with sera from histoplasmosis patients. It is very important that sensitive chemiluminescence immunoassays be used with purified antigenic molecules to ensure specificity for the diagnosis and follow‐up of parasitic and fungal infections. © 1994 Wiley‐Liss, Inc.
Enzymatic fluorometric assay for tissue cAMP
Authors: Sugiyama Atsushi , Wiegn Phi , McKnite Scott , Lurie Keith G.
Published: March 2007

cAMP is commonly measured using either immunoassay or high‐performance liquid chromatography. The current methods are sensitive but may lack versatility and be expensive; also, radioactivity is potentially harmful to the operator and environment. Given these concerns, we developed a highly sensitive enzymatic fluorometric assay for cAMP. The method consists of five steps: (1) destruction of interfering compounds with apyrase, 5′ nucleotidase, adenosine deaminase, and alkaline phosphatase; (2) conversion of cAMP to AMP; (3) conversion ofAMP to ATP; (4) amplification ofATP by ATP‐ADP cycling; and (5) fluorometric measurement of resultant NADPH. cAMP was measured in male Sprague Dawley rats anesthetized with pentobarbital. Stimulated rats (n = 4) received isoproterenol (16 μg/kg, s.q.) and aminophylline (20 mg/kg, s.q.), whereas controls (n = 4) received no additional drug. With the enzymatic fluorometric assay, cAMP content in heart, liver, and kidney (pmol/mg wet wt, mean ± SEM) was 0.34 ± 0.03, 0.33 ± 0.03, and 0.92 ± 0.11 in the control group and 0.77 ± 0.10, 0.66 ± 0.04, and 1.53 ± 0.12 in the stimulated group, respectively. The total assay duration including sample reading procedure varied at 4.5–9.5 hr, depending on its sensitivity. cAMP from the same samples was measured using a commercially available enzyme immunoassay kit and was found to be very similar to the enzymatic fluorometric assay. We conclude that this new assay is sensitive, safe, versatile, and inexpensive and can be used to measure cAMP in multiple types of tissue, including biopsy samples weighing <200 μg. © 1994 Wiley‐Liss, Inc.
Rapid method for purification of CD56+ natural killer cells with preferential enrichment of the CD56bright+ subset
Authors: Handgretinger Rupert , Welte Birgitta , Dopfer Roland , Niethammer Dietrich
Published: March 2007

A rapid method for the purification of CD56+ natural killer (NK) cells with the preferential enrichment of the CD56bright+ subset is described. The method is based on the adsorption of CD56+ cells indirectly stained with a biotinylated antibody on an avidin‐coated column. The adsorbed CD56+ cells can then be squeezed out mechanically without damaging the viability or the function of the cells. Starting from peripheral mononuclear cells from adult donors, the recovery of the CD56+ cells was 11.9 ± 9.0% (n = 8), the purity 93.0 ± 4.9% (n = 10), and the enrichment factor 7.5 ± 1.7 (n = 8). Further phenotypic classification of the CD56+ cells into CD56dim+ and CD56bright+ cells showed a preferential enrichment of the CD56bright+ phenotype with a recovery of 40.5 ± 19.6% (n = 8), a purity of 30.3 ± 13.2% (n = 8), and an enrichment factor of 29.8 ± 7.2 (n = 8). In conclusion, the described method allows the rapid purification of CD56+ NK cells with the preferential enrichment of the CD56bright+ subset. © 1994 Wiley‐Liss, Inc.
Laboratory study on occult gastrointestinal bleedings: Differential diagnostic method for the parallel study of intact and fragmented hemoglobin
Authors: Ottó Szabolcs , Németh Mária , Kocsis György
Published: March 2007

The authors produced various degradation products of human hemoglobin. With the aid of immune sera raised against native hemoglobin and their fragments, we examined the antigenicity of the intact molecule and fragments as well as the specificity and reactivity of the immune sera used. Since conventional guaiac‐type reactions make no distinction between the different hemoglobin molecules and the methods described in recent years' publications only with immune sera raised against native intact hemoglobin, the demonstration of occult fecal blood has not reached the desired rate of efficacy in clinical practice. The authors recommend the introduction of a simple differential diagnostic method suitable for the parallel examination of intact and fragmented hemoglobin.©1995 wiley‐Liss, inc.
Decreased cytoplasmic immunoglobulin a production during epstein‐barr virus immortalization on lymphocytes from patients with ataxia‐telangiectasia
Authors: Okano Motohiko , Sakiyama Yukio , Matsumoto Shuzo , Mizuno Fumio , Osato Toyoro
Published: March 2007

Previously, we have reported significantly lower immunoglobulin (Ig) A production in supernatants of cultured lymphoblastoid cells using enzyme‐linked immunosorbent assay from patients with ataxia‐telangiectasia (AT) when compared to that of age‐ and sex‐matched healthy individuals. Here, we further assess the degree of cytoplasmic Ig production in these cells and also analyze it during the early phase of Epstein‐Barr virus immortalization. All classes of cytoplasmic IgM, IgG, and IgA productions were demonstrated in cells from healthy controls. In contrast, cells from patients with AT showed only cytoplasmic IgM and IgG with low or nondetectable levels of IgA during and after the immortalizing process. These results suggest B lymphocytes bearing IgA are functionally immature and/or defective in patients with AT.©1995 wiley‐Liss, inc.
Quantification of mitogen induced human lymphocyte proliferation: Comparison of alamarbluetm assay to 3h‐thymidine incorporation assay
Authors: De Fries Ricarda , Mitsuhashi Masato
Published: March 2007

Proliferation of human lymphocytes in response to various stimuli has traditionally been assessed by measuring uptake of radiolabeled nucleotides such as 3H‐thymidine. We have evaluated a fluorometric assay, which uses the commercially available reagent, alamarBlue, as a potential substitute for the 3H‐thymidine assay in measuring proliferation of human lymphocytes. In this assay, alamarBlueTM is added to a population of cells where it is reduced by mitochondrial enzyme activity. The reduced form of the reagent is fluorescent and can be quantitatively detected. The safety and convenience of the alamarBlueTM assay make it very attractive for use in the clinical laboratory.

In this study peripheral blood mononuclear cells (PBMC) from healthy donors were stimulated using the mitogen Concanavalin A, and proliferation was assessed using either the 3H‐thymidine or the alamarBlueTM assay. The alamarBlueTM assay reliably detects human PBMC and we found that the linear range of detection was 104 cells/well (96‐well plate) to 5 × 105 cells/well. Detection of human PBMC is highly reproducible and the alamarBlue assay may be suitable in a number of applications where detection or relative quantitation of human PBMC is required. The alamarBlue assay also detected mitogen induced proliferation of PBMC although with a significantly lower level of sensitivity than the standard 3H‐thymidine assay.©1995 wiley‐Liss, inc.
Detection of (1–3)‐β‐D‐glucan in a rat model of aspergillosis
Authors: Mitsutake Kotaro , Kohno Shigeru , Miyazaki Takashige , Yamamoto Yoshihiro , Yanagihara Katsunori , Kakeya Hiroshi , Hashimoto Atsuro , Koga Hironobu , Hara Kohei
Published: March 2007

The G test containing factor G, fractioned from the Limulus lysate, was used to detect (1–3)‐β‐D‐glucan in a rat model of aspergillosis. Aspergillus fumigatus strain MF‐13, 1 × 104 conidia, were inoculated transtracheally into rats treated with cortisone acetate (100 mg/kg) and fed a low‐protein (8%) diet. Increased serum (1–3)‐β‐D‐glucan was found on the sixth day after inoculation in concentrations of 370 ± 178 pg/ml (mean ±SD) in untreated controls, and 154 ± 43 pg/ml in rats treated with 0.5 mg/kg of amphotericin B. On day 11 (1–3)‐β‐D‐glucan concentrations were 2,590 ± 2,940 pg/ml and 448 ± 442 pg/ml, respectively. The elevation in levels of (1–3)‐β‐D‐glucan increased in correlation with the elevation of galactomannan antigen titers; (1–3)‐β‐D‐glucan is thus measurable during experimental aspergillosis in rats.©1995 wiley‐Liss, inc.
Prostate‐Specific antigen expression by various tumors
Authors: Levesque Michael , Hu He , Diamandis Eleftherios P. , D'Costa Mario
Published: March 2007

There is a growing body of evidence indicating that prostate‐specific antigen (PSA) may be present in many steroid hormone‐stimulated epithelial tissues other than that of the prostate. In particular, breast tumor cell lines treated with steroid hormone receptor agonists, breast tumors, and normal human breast have recently been found by our group to contain PSA. To investigate whether PSA may also be present in other human tumors, we employed a highly sensitive immunofluorometric assay technique to quantify PSA immunoreactivity in tumor extracts. Using a PSA‐positivity cutoff value of 0.005 ng per mg of protein, 23 of 43 diverse tumors tested positive for PSA protein. Confirmatory analyses for PSA by a commercially available method (IMx) on six samples demonstrated a high degree of concordance between the two methods. To establish the molecular weight of the immunoreactive species, the most highly positive tumor extracts of each tumor type were fractionated by high performance liquid chromatography. Whereas the majority of tumors had immuoreactivity eluting at both 100 KDa and 33 KDa, corresponding to PSA bound to α1‐antichymotrypsin and free PSA, respectively, the colon and parotid tumors displayed immunoreactivity only at the 33 KDa fraction. We conclude that in addition to breast tumors and normal breast, colon, ovarian, liver, kidney, adrenal, and parotid tumors can also produce PSA. The physiological role of PSA in these tumors is currently under investigation.©1995 wiley‐Liss, inc.
Evaluation of a new automated system for the determination of ck‐mb isoforms
Authors: Secchiero Sandra , Altinier Sara , Zaninotto Martina , Lachin Mattia , Plebani Mario
Published: March 2007

We evaluated a new analyzer (Cardio REP) specifically designed for cardiac CK‐MB isoenzyme and isoforms activity, with a performance time of 24 minutes. Ten AMI patients, with times elapsed between the onset of chest pain and admission to hospital ranging from 30 minutes to 4 hours, were monitored every 3–4 hours until the 16th hour of hospitalization. In each serum sample, in addition to total CK‐MB and CK‐MB isoforms measured by the Cardio REP analyzer, we also assayed total CK activity, CK‐MB activity by immunoinhibition method, CK‐MB mass concentration, CK‐MB isoforms by REP method, troponin T, and myoglobin. The precision study demonstrated acceptable within assay and between assay CVs% for total CK‐MB (8.1 and 10.4), MB1 (9.1 and 14.2), and MB2 (9.1 and 8.2) isoforms. The method was found to be linear up to 371 U/L for MB2 isoform fraction and up to 516 U/L for total CK‐MB. Results for CK‐MB obtained with the Cardio REP correlated well with those for CK‐MB activity obtained with the immunoinhibition method (r = 0.869) and those of CK‐MB mass concentration (r = 0.923). The sensitivity of the Cardio REP CK isoforms method was found to be greater than that of the REP CK isoforms method. Time to first increased value of MB2/MB1 ratio and MB2 isoform was earlier in comparison to that for CK‐MB mass concentrations and similar to that for myoglobin, a marker that, however, lacks specificity. The diagnostic efficiency of CK‐MB isoforms and the availability of a real‐time, fully automated method for their measurement suggest the utilization of this biochemical marker in emergency for the early diagnosis of AMI.
Utilization of monoclonal‐antibody‐based assay (hemocardtm) in screening for and differentiating between genotypes of sickle cell disease and other hemoglobinopathies
Authors: Schultz John C.
Published: March 2007

Sickle cell disease covers a group of conditions in which pathology may be attributed to the presence of sickle hemoglobin (HbS). The identification of HbS and other variants including those in combination with HbS is commonly achieved by cellulose acetate electrophoresis at alkaline pH. Because many hemoglobin variants with similar charges have similar electrophoretic migration patterns, they are difficult to differentiate by electrophoresis. The HemoCardTM assays address this concern through the use of monoclonal antibodies capable of specifically recognizing the unique amino acid sub‐stitution in the variant hemoglobin. The panel of HemoCard monoclonal antibodies confirms the absence and presence of HbA, HbC, HbE, HbS, and other sickling hemoglobin variants. The combination of alkaline cellulose acetate electrophoresis and HemoCard assays allows the technologist to reach a final confirmation of both common and much less common sickle cell disease genotypes, combinations of HbS with other hemoglobins that ordinarily do not produce sickle cell disease, and other clinically important hemoglobinopathies including HbE/ß‐thalassemia and hemoglobin C disease.
Multilayer peroxidase‐labeled antibody method: Comparison with labeled streptavidin‐biotin method, avidin‐biotin‐peroxidase complex method, and peroxidase‐antiperoxidase method
Authors: Shin Masashi , Izumi Shin‐Ichi , Nakane Paul K.
Published: March 2007

A new, extremely sensitive, “multilayer peroxidase‐labeled antibody method” is introduced. This method utilizes peroxidase‐labeled antibodies that immuno‐react with their antigen, but also can be an antigen to other peroxidase‐labeled antibodies. An example of this method is: use rabbit IgG as the first antibody, peroxidase‐labeled goat antirabbit IgG as the second antibody, peroxidase‐labeled rabbit antigoat IgG as the third antibody, peroxidase‐labeled goat anti‐rabbit IgG as the fourth antibody, and peroxidase‐labeled rabbit antigoat IgG as the fifth antibody in succession. We compared the sensitivities of this multilayer peroxidase‐labeled antibody method with that of commercially available immuno‐peroxidase staining kits, i.e., avidin‐biotin‐complex method and peroxidase‐antiperoxidase method, both by immunostaining and enzyme‐linked immunosorbent assay. The generated signal and background in immunostained sections were determined by a computer‐assisted image analyzer and by enzyme‐linked immunosorbent assay spectrophotometrically. It was found that a maximum signal/background ratio was obtained with those sections reacted with the first four antibodies in succession by the multilayer peroxidase‐labeled antibody method, and those reacted with the five antibodies in succession by the multilayer peroxidase‐labeled antibody method in the enzyme‐linked immunosorbent assay. Thus, the multilayer peroxidase‐labeled antibody method is more sensitive than the avidin‐biotin‐complex method and the peroxidase‐antiperoxidase method.
Dynamic MR imaging of the gastroesophageal junction in healthy volunteers during bolus passage
Authors: Kulinna‐Cosentini C. , Schima W. , Cosentini E.P.
Published: March 2007

Purpose

To evaluate the feasibility of noninvasive dynamic fast magnetic resonance imaging (MRI) during swallowing in healthy volunteers, and to determine esophageal function at the gastroesophageal junction during swallowing.

Materials and Methods

A total of 20 healthy volunteers underwent MRI while swallowing in the supine position. Dynamic gradient‐echo (GRE) sequences (balanced fast field echo [B‐FFE]) were employed in three planes on a 1.5T unit using a phased‐array body coil. Buttermilk spiked with gadolinium (Gd) chelate (40:1) for bolus passage was used as an oral contrast agent. We evaluated visualization of esophageal bolus transit, bolus transit time (BTT), peristalsis, identification of the gastroesophageal junction, and reflux during the Valsalva maneuver.

Results

The mean visible length of the esophagus was 16.2 ± 5.3 cm in the sagittal view, and 13.8 ± 4.9 cm in the coronal view. In the sagittal view the BTT was defined in 15 of 20 volunteers and was 7.6 ± 1.4 seconds. The BTT in the coronal view was measured in seven of 20 volunteers and was 8 ± 1.3 seconds on average. The axial view yielded higher scores (2.25) than the coronal (1.98) and sagittal (1.78) views for identification of the cardia and during the Valsalva maneuver. Bolus contrast was better displayed in the sagittal (2.2) view than in the coronal (2.08) or axial (1.73) planes. In six volunteers, gastroesophageal abnormalities, such as axial hernia, reflux, and nonperistaltic contractions, were identified. For statistical analysis we used the Friedman test and a one‐way analysis of variance (ANOVA).

Conclusion

The results indicate that dynamic MR swallowing is a feasible and reproducible technique that warrants further studies in patients. J. Magn. Reson. Imaging 2007. © 2007 Wiley‐Liss, Inc.
Embryonic stem cells produce neurotrophins in response to cerebral tissue extract: Cell line‐dependent differences
Authors: Bentz Kristine , Molcanyi Marek , Riess Peter , Elbers Andrea , Pohl Esther , Sachinidis Agapios , Hescheler Jürgen , Neugebauer Edmund , Schäfer Ute
Published: March 2007

In the present study, we compare the capacity of two different embryonic stem (ES) cell lines to secrete neurotrophins in response to cerebral tissue extract derived from healthy or injured rat brains. The intrinsic capacity of the embryonic cell lines BAC7 (feeder cell‐dependent cultivation) to release brain‐derived neurotrophic factor (BDNF) or neurotrophin‐3 (NT‐3) exceeded the release of these factors by CGR8 cells (feeder cell‐free growth) by factors of 10 and 4, respectively. Nerve growth factor (NGF) was secreted only by BAC7 cells. Conditioning of cell lines with cerebral tissue extract derived from healthy or fluid percussion‐injured rat brains resulted in a significant time‐dependent increase in BDNF release in both cell lines. The increase in BDNF release by BAC7 cells was more pronounced when cells were incubated with brain extract derived from injured brain. However, differences in neurotrophin release associated with the origin of brain extract were at no time statistically significant. Neutrophin‐3 and NGF release was inhibited when cell lines were exposed to cerebral tissue extract. The magnitude of the response to cerebral tissue extract was dependent on the intrinsic capacity of the cell lines to release neurotrophins. Our results clearly demonstrate significant variations in the intrinsic capability of different stem cell lines to produce neurotrophic factors. Furthermore, a significant modulation of neurotrophic factor release was observed following conditioning of cell lines with tissue extract derived from rat brains. A significant modulation of neurotrophin release dependent on the source of cerebral tissue extract used was not observed. © 2007 Wiley‐Liss, Inc.
Increased levels of fucosyltransferase IX and carbohydrate Lewisx adhesion determinant in human NT2N neurons
Authors: Brito Catarina , Escrevente Cristina , Reis Celso A. , Lee Virginia M.‐Y. , Trojanowski John Q. , Costa Júlia
Published: March 2007

The expression of the fucosylated carbohydrate Lewisx (Lex) determinant (Gal(β1–4)[Fuc(α1–3)]GlcNAc‐R) has been found in glycoproteins, proteoglycans, and glycolipids from the nervous system. Evidence suggests its association with cell–cell recognition, neurite outgrowth, and neuronal migration during central nervous system development. In the present work, we detected increased levels of Lex in differentiated human NT2N neurons cultured in vitro. To identify which fucosyltransferase (FUT) synthesized the Lex in NT2N neurons, RT‐PCR, FUT substrate specificity and Western blot analysis were carried out. Strong activity toward acceptors Galβ4GlcNAc‐O‐R and Fucα2Galβ4GlcNAc‐O‐R [R = ‐(CH2)3NHCO(CH2)5NH‐biotin], together with strong FUT9 detection by Western blot and presence of transcripts showed that FUT9 was the enzyme associated with Lex biosynthesis in NT2N neurons. Lex was detected at the plasma membrane of NT2N neurons, in lysosomes marked with lysosomal‐associated membrane protein 1 (LAMP‐1), and it was found for the first time to colocalize with the tetanus neurotoxin‐insensitive vesicle‐associated membrane protein (TI‐VAMP) that defines the TI‐VAMP exocytic compartment that is involved in neurite outgrowth. Furthermore, incubation with anti‐Lex monoclonal antibody L5 led to impaired adhesion of NT2N neurons to the surface matrix and inhibited neurite initiation. In conclusion, FUT9 and its product Lex are detected specifically in human NT2N neurons and our results indicate that they underlie cell differentiation, cell adhesion, and initiation of neurite outgrowth in those neurons. © 2007 Wiley‐Liss, Inc.
Plasma concentrations and dietary requirements of leucine, isoleucine and valine: Studies with the young chick
Authors: D'Mello J. P. Felix
Published: March 2007

The minimal requirements of the chick for leucine, isoleucine and valine have been assessed using diets based on wheat, groundnut meal and crystalline amino acids. The results of these studies clearly indicate that a relatively low isoleucine concentration of 0.52% allows satisfactory growth rate and efficiency of food conversion with the dietary levels of leucine and valine maintained at 0.98 and 0.63%, respectively. No beneficial response occurs to further supplementation with isoleucine. However, a higher isoleucine level (0.76 %) augments the requirements of leucine and valine to 1.10 and 0.75%, respectively, as judged by growth and efficiency of food conversion responses. These patterns of interactions are reflected in the plasma concentrations of leucine, isoleucine and valine.
Studies on nucleotides and related compounds in plants. V. Isolation and identification of bracken nucleotides
Authors: Duncan H. J.
Published: March 2007

The presence of acid‐soluble nucleotides was demonstrated in bracken rhizomes. Cold trichloroacetic acid extracts of bracken had to be submitted to some eight purification steps prior to analysis. Of the nucleotide material held by an anion exchange resin uridine derivatives (UMP, UDP‐glucose, UDP‐galactose, UDP‐xylose, UDP‐arabinose, UDP‐glucuronic acid and a trace of UDP) accounted for some 80% of the total. The remainder comprised guanosine (12%; GMP, GDP‐mannose and GDP‐fucose) and adenosine (8%; mainly ATP) compounds.

A comparison of the sugar composition of the nucleotide material with that of associated soluble carbohydrates present in the rhizome (mucilage, glycoside and sucrose) is also included.
Testicular development, androstenone levels and androstenone odour of untreated and trenbolone implanted boars
Authors: Ventanas Jesús , Sancho Gabriel , Garcia‐Regueiro Jose A , Antequera Teresa , Martinez Margarita , López‐Bote Clemente
Published: March 2007

Twelve boars were implanted at 50 kg live weight with 300 mg of trenbolone acetate (TBA), and 12 male litter mates were held as controls. All animals were slaughtered at 175 days of age and ∼ 100 kg live weight. TBA administration led to a decrease in testicle size (P<0.001) and testosterone levels (P<0.05). Histological evidence of modifications of the intertubular area (Leydig cells) in the testicle was found. Decrease of weight and endocrine secretion in the testes reduced the accumulation of 5α‐androstenone in adipose tissue from a mean ± SEM value of 0–52 ± 0–16 μg g−1 fat for untreated boars to 0–22±0–04 μg g−1 fat for implanted animals. Sensory evaluation of androstenone odour using the hot iron method demonstrated that the percentage of unfavourable opinions was high for control boars whereas none of the implanted males exhibited detectable androstenone odour. These induced modifications of testicular secretion by androgen administration in pigs—in contrast to castration—have the advantage of reducing the androstenone odour without decreasing the productive performance of entire males.
A sandwich enzyme‐linked immunosorbent assay for κ‐carrageenan determination
Authors: Arakawa Shoichi , Ishihara Hideko , Nishio Osamu , Isomura Shin
Published: March 2007

High titre antisera to κ‐carrageenan were produced by immunisation of rabbits and guinea pigs with K‐carrageenan. These antisera, rabbit serum as a capture antibody and guinea pig serum as a detector antibody, were used in a sandwich enzyme‐linked immunosorbent assay. The range of determination of K‐carrageenan was from 16 to 256 ng ml−1 in the assay. The antisera showed no cross‐reactivity with six thickeners. The assay was developed for application to foods such as jelly and custard pudding and demonstrated many advantages including high sensitivity, technical simplicity and the minimal requirement for food sample preparation before assay.
Development of a rapid colorimetric method for the determination of chlorogenic acid in freeze‐dried potato tubers
Authors: Griffiths D Wynne , Bain H , Dale M F B
Published: March 2007

After‐cooking blackening (ACB) in potato cultivars results from the formation of complex compounds of chlorogenic acid and iron ions. At present the estimation of chlorogenic acid content represents the most reliable method for predicting the extent of ACB in material in a potato breeding programme to allow subsequent selection. A number of methods for determining chlorogenic acid levels, including high performance liquid chromatography, sodium nitrite and other colorimetric and chromatographic assays, are evaluated. A rapid screening method for the direct determination of chlorogenic acid levels is described, and its application within a breeding programme and the results obtained from a number of cultivars are discussed.
Purification and partial characterisation of trypsin inhibitors from seeds of Clitoria ternatea
Authors: Macedo Maria Ligia R , Xavier‐Filho José
Published: March 2007

Three trypsin inhibitors of 20, 12 and 7 kDa were isolated in pure form from seeds of the vine Clitoria ternatea L of the tribe Glycineae of the Papilionoideae. The 20‐kDa inhibitor is a one‐chain molecule with arginine in the reactive site. The 12‐kDa one is also a one‐chain molecule and has lysine in the reactive site. It is also an inhibitor of chymotrypsin. They were tentatively assigned to the Kunitz and Bowman‐Birk families, respectively. The small molecular weight inhibitor (7 kDa) also has an arginine in the reactive site and is probably of the Bowman‐Birk type.
The mechanism of drip production: Formation of two compartments of extracellular space in muscle Post mortem
Authors: Offer Gerald , Cousins Tony
Published: March 2007

The structural changes occurring post mortem at 10°C in beef sternomandibularis muscle have been studied by fixing muscles at various times post mortem and examining transverse sections by light microscopy, or transversely cut surfaces by scanning electron microscopy.

At 2 h post mortem, the muscle fibres filled the endomysial network and the fibre bundles filled the perimysial network. At 4 to 6 h post mortem, presumably as a result of fibre shrinkage, gaps appeared between the fibre bundles and the perimysial network but at this stage the fibres still filled the endomysial network. Gaps between fibres did not appear until about 24 to 48 h post mortem.

It is concluded that at rigor onset, when myofibrils shrink owing to pH fall and the attachment of myosin heads to actin filaments, the fluid expelled accumulates in these two types of extracellular space. Such fluid, especially that accumulating between fibre bundles, is the likely source of drip.
Comparative studies of the degradation of non‐starchy polysaccharides by sorghums and barleys during malting
Authors: Etokakpan Okokon U
Published: March 2007

The β‐D‐glucan of whole grains of four sorghum (Sorghum bicolor (L) Moench) varieties grown in Nigeria, unlike those of barley, is not significantly reduced during malting. Pentosan level, while enhanced in barley (Hordeum rulgare L) during the same period, dropped in sorghum. Apparently sorghums have very low β‐D‐glucan degrading endo β 1–3,1‐4 glucanase activity as observed from their failure to hydrolyse their isolated endosperm cell walls and barley β‐D‐glucan extracted in water at 40°C.
Flavonoids in three olive (Olea europaea) fruit varieties during maturation
Authors: Vlahov Giovanna
Published: March 2007

The objective of the present paper was the characterisation of olive fruit varieties through the formation and variation during maturation of their flavonols, flavones and anthocyanins. An olive variety with a constant cyanidin‐3‐rutinoside:cyanidin‐3‐glucoside ratio of 1 exhibited lower contents of luteolin‐7‐glucoside and quercetin‐3‐rutinoside than varieties in which the only anthocyanin detected was cyanidin‐3‐rutinoside.
Particle size distribution and solubility of dietary fibre in swede‐ (Brassica napus) based and wheat‐bran‐based diets during gastrointestinal transit in the pig
Authors: Robertson James A , Murison Sandra D , Chesson Andrew
Published: March 2007

Changes in fibre solubility, particle size and distribution and microbial activity during gut transit in pigs fed either a swede‐based or wheat‐bran‐based diet were monitored. Digesta samples recovered from the stomach, ileum and caecum, together with feed and faecal samples, were examined for changes in the chemical and physical properties of the fibre fraction. Solubility of swede fibre was increased in the upper gut where a fourfold reduction in particle size and change in particle size distribution also occurred. This was associated with a loss of pectic polysaccharide. Little change in bran properties was noted in the foregut although the shape of bran particles was modified in the large intestine. Overall loss of the fibre fraction during gut transit was 41‐5% for bran and 87‐8% for swede. Microbial colonisation of the upper gut was significant on both diets. Chemical conditions during transit were thought to predispose pectic polysaccharides to solubilisation, and to encourage tissue maceration and hence particle size reduction. Lack of pectic polysaccharides in wheat bran and/or lignification maintained particle integrity in this diet. These properties may be important in influencing the physiological response to fibre in the diet.
Sterols of seed oils of Vernonia galamensis, Amaranthus cruentus, Amaranthus caudatus, Amaranthus hybridus and Amaranthus hypochondriacus grown in the humid tropics
Authors: Ologunde M O , Ayorinde F O , Shepard R L , Afolabi O A , Oke O L
Published: March 2007

Analysis of sterols of seed oils from Vernonia galamensis, Amaranthus cruentus, A caudatus, A hybridus and A hypochondriacus, the last four being exotic breeds planted in the humid tropics of Africa, is presented in this report. Identifiable sterols in all the seed oil samples include campesterol, stigmasterol, ß‐sitosterol, Δ5‐avenasterol and Δ7‐avenasterol except for Vernonia galamensis where cholesterol was detected (63.61 mg per 100 g oil).
Modelling the aqueous extraction of soluble substances from ground roasted coffee
Authors: Zanoni Bruno , Pagliarini Ella , Peri Claudio
Published: March 2007

This work aims to study and model the extraction of soluble substances from coffee grounds having different particle sizes. The results demonstrate that coffee brewing cannot be considered a simple diffusion mechanism as reported in the literature. The phenomenon has two phases: a washing phase where surface soluble substances instantaneously dissolve and are transferred to the extract by a convection mechanism, and a diffusion phase involving the solubles within the porous particles. A model was set up to calculate extraction yield as the addition of the two phases.

It was also demonstrated that the effect of washing shows a linear correlation with the external surface area of particles and that the composition of the extracts does not depend on extraction time.
Novel volatile thermal degradation products of thiamine
Authors: Ames Jennifer M , Hincelin Odile , Apriyantono Anton
Published: March 2007

Thirteen compounds (nine thiophenes, two alicyclic sulphur compounds and two aliphatic hydroxyketones) are reported as novel thermal degradation products of thiamine. Most possessed meaty and/or onion‐like aromas.
Chromatographic and non‐chromatographic characterization of poly‐α‐olefins
Authors: Voelkel Adam , Fall Jacek
Published: March 2007

Both chromatographic and non‐chromatographic techniques were used in the characterization and identification of poly‐α‐olefins. Synthetic base oils, although produced from the same raw material, exhibit different physico‐chemical properties. Their mutual miscibility and behaviour in final engine oils may be predicted from data collected through classical physico‐chemical analysis, simulated distillation chromatography as well as inverse gas chromatography. Copyright © 2007 John Wiley & Sons, Ltd.
Conversion coating treatment for AZ91 magnesium alloys by a permanganate‐REMS bath
Authors: Liang C.H. , Zheng R.F.
Published: March 2007

A permanganate‐rare earth metal salt (REMS) chemical conversion bath was applied to a sample of AZ91 magnesium alloy in this study, a red‐brown conversion coating formed subsequently on the sample surface. The test results of this coating with a scanning electron microscope (SEM) showed that there existed net‐like cracks on the surface of the treated magnesium alloy. With the analyses of X‐ray Diffraction (XRD) and X‐ray Photoelectron Spectroscopy (XPS), a further study of this coating indicated that the coating was structurally amorphous and mainly composed of CeO2, MnO, MnO2, MgO, Mg(OH)2 and MgAl2O4. Furthermore, the electrochemical polarization tests showed that compared with the samples treated by the chrome‐based method, the open‐circuit potential of the magnesium alloy coated in permanganate‐REMS bath moved from − 1.34 VSCE to − 1.28 VSCE and the anodic current density of the alloy, at the same potential, decreased evidently in simulated sweat fluid. The cracks in the chemical conversion coating should be caused by the phase structure of the magnesium alloy. During the chemical conversion process, the localized corrosion micro‐cell led to the formation of the net‐like cracks on the surface. Simultaneously, the dehydration of the surface coating after treatment also accelerated the formation of the cracks at the coating surface.
Vascularized fibular graft for bone reconstruction of the extremities after tumor resection in limb‐saving procedures
Authors: Minami Akio , Kutsumi Keiji , Takeda Naoki , Kaneda Kiyoshi
Published: March 2007

We treated 18 patients (8 males and 10 females) with bone or soft tissue tumors in the extremities by vascularized fibular grafts (VFGs). The average age was 29 years. Two had malignant soft tissue tumors in the forearm (angiosarcoma and synovial sarcoma) and the other 16 had bone tumors [osteosarcoma (8), chondrosarcoma (3), angiosarcoma (1), adamantinoma (1), and giant cell tumor (3)]. Affected sites of the bone tumor cases were tibia (5), femur (5), humerus (3), radius (2), and ulna (1). According to the surgical staging system of Enneking et al. 1 patient was in stage III, 12 in IIB, 2 in IB, and 3 in IA. The surgical margin was curative in 5 patients, wide in 9, marginal in 2, and intralesional in 2. One patient died due to lung metastasis although bone union was obtained by the VFGs. Functional results of the bone reconstruction in the remaining 17 patients were evaluated according to the modified scale of Enneking et al. The VFGs resulted in substantial bone unions in all patients except 1. Bone unions in all 17 patients occurred in less than 10 months. Overall clinical results were satisfactory. Based on a 30‐point scale overall clinical results ranged from 10 to 30 points with an average of 21 points. Functional evaluation in the upper extremity were, generally, superior to those in the lower extremity. Postoperative complications were 5 metastases (4 in the lung and 1 in the mediastinum) in 4 patients, 1 recurrence, and 2 fractures after obtaining bone union. One patient died from metastasis. These results suggest that the VFG is effective for salvaging limb function where a massive bone defect results from wide resection of tumor in the extremities. © 1995 Wiley‐Liss, Inc.
The rat brachial plexus and its terminal branches: An experimental model for the study of peripheral nerve regeneration
Authors: Bertelli Jayme Augusto , Taleb Madgid , Saadi Assia , Mira Jean‐Claude , Pecot‐Dechavassine Monique
Published: March 2007

Despite the introduction of microsurgical techniques into clinical practice, the results of surgical procedures involving the brachial plexus and peripheral nerves are still far from spectacular. We therefore studied the rat brachial plexus and its terminal branches in 203 rats. Detailed anatomic and morphologic analyses of the biceps brachii and musculocutaneous nerve, finger flexors, flexor carpi radialis, and the median nerve were performed. Various sources of conventional and vascularized nerve grafts were explored. After musculocutaneous nerve section or median nerve section, there were no articular contractures or automutilations, which constitutes an advantage for these experimental models over the sciatic nerve model. The brachial plexus and its terminal branches provide a good experimental model which can be used to assess the development and normal control of muscle function, examine the mechanisms underlying functional recovery, and test the effects of treatments to enhance recovery. © 1995 Wiley‐Liss, Inc.
Experimental microvenous thrombosis following anastomosis with a knotted suture
Authors: Tonken Heidi P. , Zhang Feng , Yim Kenneth K. , Gencosmanoglu Reha , Sudekum Anthony E. , Campagna‐Pinto Dante , Newlin Leonard , Buncke Harry J. , Lineaweaver William C.
Published: March 2007

An experimental model for producing venous thrombosis was developed in end‐to‐end anastomoses of femoral veins in rats. The anastomoses were performed using a suture (9–0 suture‐70 μm needle) with a knot 1 cm from the needle. The knot was formed by making either six or eight half‐hitches in one throw of the suture. Vessel patency was assessed through the direct “milking test” at 20 min and 24 hrs. The incidence of thrombosis when using one knot with six half‐hitches ranged from 20% to 40% and with eight half‐hitches, from 50% to 70%. The incidence of femoral vein thrombosis varied directly with the presence and size of the knot. In this model, thrombosis was induced by exaggerating vessel injuries that may occur when performing routine microvascular anastomoses. This study demonstrates a reproducible thrombogenic model which mimics clinical practice and may be used to study the effects of local and systemic antithrombogenic agents. © 1995 Wiley‐Liss, Inc.
Development of a mouse aortic transplant model of chronic rejection
Authors: Koulack Joshua , McAlister Vivian C. , Giacomantonio Carman A. , Bitter‐Suermann Hinrich , McAdonald Allan S. , Lee Timothy D. G.
Published: March 2007

Chronic rejection is the most common cause of late graft failure after solid organ transplantation. A model of chronic rejection, the rat aortic allograft, has histologic features that parallel those in the vessels of human transplanted organs. However, the molecular tools required to dissect the immunology of chronic rejection are unavailable in the rat. We developed aortic transplantation in the mouse as a new model of chronic rejection. This will allow the use of the diversity of recombinant cytokines and monoclonal antibodies available for the mouse and its well‐defined genetics to investigate chronic rejection in greater detail. We describe the perioperative care and surgical technique for the model in which a 1 cm segment of donor thoracic aorta was used to replace a section of recipient abdominal aorta below the renal arteries and above the aortic bifurcation. Mortality rates were initially high (70%) due to thrombosis and shock. Changes in technique and operator facility resulted in a high rate of success (75%). After 192 operations, the current success rate is >80%. Mice free from complications at 12 hrs postop had indefinite survival, and after 2 months the typical vascular lesion of chronic rejection was present. This new model of chronic rejection will be a valuable tool to study the molecular immunology and genetics of chronic rejection. © 1995 Wiley‐Liss, Inc.
Clinical use of microvascular PTFE grafts
Authors: Lanzetta Marco
Published: March 2007

This paper reports on the first clinical case where a polytetrafluoroethylene (PTFE) graft of less than 3 mm in diameter was implanted in the arterial system of a patient to bridge a vascular defect. A 1.5 cm long, 1.5 mm diameter prosthesis was interposed in the superficial palmar arch of a man who sustained a laceration of the palm of his dominant hand. The graft was implanted by means of the 3M Precise Microvascular Anastomotic System on one end, and by conventional microsurgical technique on the other end. There were no postoperative complications. The patient resumed his preinjury activities 9 weeks after trauma. Serial Doppler ultrasound examinations showed normal blood flow and complete patency of the graft. An angiogram performed 12 weeks postoperatively confirmed the patency of the prosthesis. Twelve months post‐operatively the patient is free from complications. The use of small diameter PTFE prostheses may be considered when planning grafting procedures for microvascular defects. © 1995 Wiley‐Liss, Inc.
Comparison of nylon and polypropylene sutures in a microvenous thrombosis model
Authors: Üstüner Tuncay E. , Zhang Feng , Lineaweaver William C.
Published: March 2007

Thrombosis rates of femoral end‐to‐end microvenous anastomoses with nylon and polypropylene sutures (9–0 suture, 70 μ needle) were compared in a microvenous thrombosis model. The vessel injuries were produced during anastomosis by using a suture with a knot 1 cm from the needle. Anastomotic thrombosis rates were assessed by visual inspection and strip test at 24 hr postoperatively. Low thrombosis rates (0% and 20%, respectively) of anastomoses with both unknotted nylon and unknotted polypropylene sutures were obtained. Anastomoses with knotted nylon and polypropylene sutures resulted in 65% and 45% thrombosis rates, respectively. Statistical analyses showed that there were no significant differences between thrombosis rates, both in anastomosis performance with unknotted nylon and polypropylene sutures (P > 0.05) and with knotted nylon and polypropylene sutures (P > 0.05), while there were significant differences between the thrombosis rate using unknotted sutures and knotted sutures (P<0.01). It was concluded that vascular injury, not suture material, is the main factor leading to thrombosis in this model. © 1995 Wiley‐Liss, Inc.
Application of sciatic functional index in nerve functional assessment
Authors: Shen Ninjiang , Zhu Jiakai
Published: March 2007

In order to confirm the reliability of the sciatic functional index (SFI) in the rat, SFI, muscle strength, electrophysiological, and morphometric assessments were carried out from the 10th day to the sixth month after nerve injury or repair. The results showed that the SFI has a positive correlation with all tested indices of muscle strength, electrophysiology and morphology (r = 0.925–0.996, P<0.01 or P<0.001). These results indicate that the SFI is a reliable index for evaluating rat sciatic nerve regeneration and can be widely used. © 1995 Wiley‐Liss, Inc.
Comparison of the telescoping anastomotic technique with the end‐to‐end technique utilizing vein grafts for venous defects: Short‐ and long‐term results
Authors: Saitoh Satoru , Kitagawa Eiji , Nakatsuchi Yukio
Published: March 2007

The telescoping anastomotic technique was used for both anastomoses of vein grafts placed in the rat left epigastric vein in an effort to make the anastomosis easier and faster. Vein grafting in the right epigastric vein was completed using the conventional technique. The 20 rats with 20 left and 13 right vein grafts, which were patent at 3 weeks, were seen again 3 months postoperatively to compare the two techniques with regard to long‐term stenosis and histological changes. Patency of the grafts 2 hr postoperatively was also compared between the two techniques in another 25 rats to eliminate the influence of recanalization on patency. All the vein grafts, which had been patent 3 weeks postoperatively, remained patent after 3 months and no statistically significant difference was found between the telescoping and conventional technique in the degree of stenosis. The vein grafts 2 hr postoperatively were all patent in both groups. © 1995 Wiley‐Liss, Inc.