bluuebunny/crossref_metadata_2025 huggingface dataset:
Detection of antibody IgG to HIV‐1 in urine by ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant p24 as antigen for diagnosis of HIV‐1 infection
Authors: Hashida Seiichi , Hashinaka Kazuya , Hirota Kouichi , Saitoh Atsushi , Nakata Atsuo , Shinagawa Hideo , Oka Shinichi , Shimada Kaoru , Mimaya Jun‐Ichi , Matsushita Shuzo , Ishikawa Eiji
Published: March 2007

Anti‐HIV‐1 IgG in urine was detected by an ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant p24 gag protein (p24) of HIV‐1 as antigen and β‐D‐galactosidase from Escherichia coli as label. Anti‐HIV‐1 IgG in urine was reacted simultaneously with 2, 4‐dinitrophenyl‐bovine serum albumin‐recombinant p24 conjugate and recombinant p24‐β‐D‐galactosidase conjugate. The complex formed, consisting of the three components, was trapped onto polystyrene balls coated with affinity‐purified (anti‐2, 4‐dinitrophenyl group) IgG, eluted with ϵN‐2, 4‐dinitrophenyl‐L‐lysine, and transferred to polystyrene balls coated with affinity‐purified (anti‐human IgG γ‐chain) IgG. Bound γ‐D‐galactosidase activity was assayed by fluorometry. This assay was at least 3, 000‐fold more sensitive than conventional methods. The lowest signal among 49 asymptomatic carriers was 3.1‐fold higher than the highest nonspecific signal among 100 seronegative subjects. The sensitivity and specificity were both 100%. The positivity could be confirmed by preincubation of urine samples with excess of the antigen. Thus, this assay would be a powerful tool for detecting IgG antibody to HIV‐1 in urine. © 1994 Wiley‐Liss, Inc.
Detection of candida enolase antibody in patients with candidiasis
Authors: Mitsutake Kotaro , Kohno Shigeru , Miyazaki Takashige , Miyazaki Haruko , Maesaki Shigefumi , Koga Hironobu
Published: March 2007

The serodiagnostic value of candidaenolase antibody was evaluated in 27 patients with systemic candidiasis. The glycolytic enzyme enolase was prepared from Candida albicans IFM 40009 and used as the antigen for immunoblot assays. IgG class antibody was found in the sera of 17 of 27 patients (62.9%) obtained during the early course of the infection. Serial sampling of sera increased the number of patients with positive Candida enolase antibody, which was detected in 25 of 27 patients (92.5%). None of the patients with localized candidiasis showed a positive IgG titer more than 1:100. The specificity of Candida enolase IgG antibody was 95%. This antibody was observed in various candida species: Candida albicans (6/7, 85.7%); C. parapsilosis (7/9, 77.9%); C. tropicalis (5/5, 100%); C. guilliermondii (4/4, 100%); and C. glabrata (1/3, 33.3%). © 1994 Wiley‐Liss, Inc.
Reactivity of chagasic sera with crude and highly purified glycosphingolipid fractions from Trypanosoma cruzi epimastigotes
Authors: Boas Maria Helena Villas , da Silva Maria Cleide Florentino , de Oliveira Tereza Gomes , Travassos Luiz Rodolpho , Bergter Eliana Barreto
Published: March 2007

The reactivities of sera from patients with Chagas disease or from T. cruzi‐immunized rabbits with two different lipid preparations of T. cruzi were assessed using epimastigote antigens. Serum reactivities were determined using a quantitative enzyme‐linked immunosorbent assay (ELISA). Antigen 1 represents the lower phase obtained from crude lipid extract after Folch partition (LCL). Antigen 2 is a highly purified glycosphingolipid fraction (GSL). The LCL antigen discriminated quite well the reactivities of Chagasic patients' sera and sera from healthy individuals, as well as between the serum from a T. cruzi‐immunized rabbit (TIRS) and normal rabbit serum (NRS). A strong reactivity with GSL was obtained with TIRS. Reactivity with GSL was also obtained with human Chagasic sera. Compared to a group of normal individuals, the reactions of antibodies directed against lipid antigens were considerably increased in sera of patients with Chagas disease. Chagasic sera did not differentiate between glycolipids with terminal β‐glucosyl or β‐galactosyl nonreducing units. They discriminated, however, glucosylceramides with differences in the ceramide structure. To determine the specificity of Chagasic sera, antibodies isolated on LCL‐immunosorbent (LCL‐Ch Abs) as well as on laminin‐immunosorbent (Lam‐Ch Abs) were tested against laminin and LCL antigens. We found that Lam‐Ch Abs reacted with murine laminin, whereas the reaction was negative with LCL. In contrast, the LCL‐Ch Abs reacted either with LCL antigens or with laminin. The reactivity with laminin was strong in comparison with LCL. Results suggest that although the glycolipids are recognized by Chagasic patients' sera, the reactive antibodies are not specific since they reacted quite well with murine laminin. Polyspecific antibodies from Chagasic sera have already been described using other unrelated antigens. © 1994 Wiley‐Liss, Inc.
Anti‐Glycosphingolipid autoantibodies in rheumatologic disorders
Authors: Zeballos Roberto S. , Fox Robert I. , Cheresh David A. , McPherson Richard A.
Published: March 2007

Antibodies directed against ganglioside GM1 or sulfatides are frequently associated with motor or sensorimotor neuropathies. To establish the prevalence of such antiglycosphingolipid autoantibodies in autoimmune disorders and to determine whether they contribute to neurologic symptoms in those individuals, we measured these antibodies by enzyme‐linked immunosorbent assay (ELISA) in serum samples from rheumatologic patients with and without peripheral neuropathies (PN). We tested 21 patients with systemic lupus erythematosus (9 with PN), 26 with Sjögren's syndrome (12 with PN), 34 with scleroderma (28 with PN), and 14 with rheumatoid arthritis (4 with PN).

Samples from 32 normal individuals were also tested. Patients with systemic lupus erythematosus and rheumatoid arthritis had elevated concentrations of GM1 antibodies and scleroderma patients had lower levels of sulfatide antibodies compared to healthy individuals. The presence of ganglioside or sulfatide antibodies did not correlate with the development of peripheral neuropathy in these patients. These findings suggest that relatively low‐titer glycosphingolipid antibodies may arise as part of a nonspecific polyclonal gammopathy in rheumatologic disorders but generally without clinical manifestation. © 1994 Wiley‐Liss, Inc.
Comparison of three commercially available enzyme immunoassays for the screening of autoantibodies to extractable nuclear antigens
Authors: Jaskowski Troy D. , Schroder Carl , Martins Thomas B. , Mouritsen Lars , Hill Harry R.
Published: March 2007

The initial screening test used in the diagnosis of connective tissue diseases is based on the detection of antinuclear antibodies (ANA) by indirect immunofluorescence (IFA). When the ANA screen is positive, it is often useful to determine the specificity of the autoantibody to a series of extractable nuclear antigens (ENA), a procedure that has been classically performed by double immunodiffusion. Testing large numbers of clinical specimens for autoantibodies to ENA by double diffusion techniques can be time‐consuming and expensive. ENA screening systems that employ enzyme immunoassay (EIA) technology have recently become commercially available. We compared three EIA ENA assays to classic Ouchterlony double diffusion techniques. Furthermore, the sensitivity of each antigen and methodology (including ANA immunofluorescence using HEp‐2 cells) was tested using ENA positive sera possessing single autoantibodies. Two of the three EIAs that detected immunoglobulin G type autoantibodies to Smith (Sm), ribonucleoprotein (RNP), Sjögren's syndrome‐associated antigens Ro (SSA) and La (SSB), were provided by INOVA and Sigma Diagnostics. A third EIA, which also included scleroderma‐associated antigen 70 (SCL‐70/DNA‐topoisomerase I) and histidyl‐tRNA synthetase (Jo‐1) in addition to the four previously stated antigens, was provided by Clark Laboratories. This latter ENA screen detected IgG, IgA, and IgM type autoantibodies. Included in the study were sera covering a wide variety of anti‐nuclear and other autoantibodies. Sensitivity was 100% for all EIA ENA screens when compared to Ouchterlony double diffusion and specificity exceeded 95% in each case. Sensitivity studies showed Ouchterlony to be less sensitive than EIA when detecting low levels of autoantibodies to ENA. After the detection of ANA antibody by IFA, EIAs utilizing combined antigens can be used to screen large numbers of clinical specimens for autoantibodies against SSA, SSB, Sm, RNP/SCL‐70, and Jo‐1 prior to confirmation by Ouchterlony double diffusion.©1995 wiley‐Liss, inc.
Criteria for consistent and high sensitivity of dna in situ hybridization on paraffin sections: Optimal proteolytic enzyme digestion
Authors: Lu Qi Long , Lawson Paul , Alero Thomas J.
Published: March 2007

It is technically challenging for the detection of target DNA in low abundancy, such as viral DNA sequences in latently infected cells by nonisotopic in situ hybridization (ISH). Consistent result is even more difficult to achieve on routine paraffin sections. Proteolytic enzyme digestion is most critical for both consistency and sensitivity of the technique. We here have investigated the effect of enzyme digestion on cell morphology, protein and DNA reduction, and hybridization efficiency. The results demonstrated that enzyme digestion improves efficiency of ISH through a process involving partial DNA purification on sections. There is a clear relationship between proteolytic enzyme digestion, morphology changes, and hybridization efficiency. Although detection of DNA sequences in abundance can be achieved within a relatively wide range of digestion levels, maximum hybridization efficiency was always related to the cells, which showed morphology of nuclear swollen, weak homogeneous chromatin staining with hematoxylin and loss of visible nuclear membrane. Detection of viral DNA in low copy number critically depends on the creation of the morphologic changes by enzyme digestion. The morphological changes would therefore serve as important criteria for optimal digestion, result interpretation, and comparison.
Comparison of elisa and hplc for the determination of desmosine or isodesmosine in aortic tissue elastin
Authors: Osakabe Toru , Seyama Yoshiyuki , Yamashita Saburo
Published: March 2007

We developed a rapid and simple method for estimating tissue elastin content by measuring desmosine (D) in tissue hydrolysates by competitive ELISA. We compared the ELISA previously reported HPLC methods. When D or isodesmosine (ID) in hydrolysate of the same elastin preparation were measured by the two different methods, a good linear relationship was obtained (r = 0.854 for human aorta or r = 0.938 for rabbit aorta, respectively). The ELISA method can detect as little as 6 pmol/ml and it may be useful in monitoring elastin metabolism in patients with various connective tissue diseases.
Dot‐Elisa for the rapid detection of gentamicin in milk
Authors: Ara Jahan , Gans Zeev , Sweeney Ray , Wolf Benjamin
Published: March 2007

A dipstick dot‐ELISA for the detection of gentamicin in milk of dairy cattle is reported for the first time. The test is based on a sandwich ELISA using high affinity monoclonal antibodies to gentamicin. Antibodies were adsorbed to nitrocellulose filters, blocked, dried, and stored for several weeks before use. The dipstick ELISA detected gentamicin at a concentration of 0.1 m̈g/ml and produced strongly positive results at 0.2 m̈g–0.3 m̈g/ml. This ELISA is highly specific and no false positives were detected when tested against various aminoglycoside analogs including streptomycin, kanamycin, bekanamycin, amikacin, neomycin, and tobramycin. Further, the elimination in cow milk of gentamicin residues following intramammary administration of the drug was studied in two dairy cattle using dot‐ELISA. Milk gentamicin levels were detected at post injection hours up to 120 hr in each of the two dairy cattle. It therefore, appears that gentamicin residues can still be detected in milk after 5 days using dot‐ELISA. Based on the simplicity of performance and the economical nature of the test system, dipstick is recommended as a suitable method for wide scale use in field studies and diagnostic laboratories.
A combined indirect elisa and immunoblotting for the detection of intrathecal herpes simplex virus igg antibody synthesis in patients with herpes simplex virus encephalitis
Authors: Markoulatos Panayotis , Labropoulou Vassiliki , Kordossi Aphroditi , Krikelis Vassilis , Spyrou Niki , Moncany Maurice L. J.
Published: March 2007

A combined indirect ELISA and immuno‐blotting assay was used for the detection of intrathecal synthesis of IgG antibodies to herpes simplex virus (HSV) in patients with HSV encephalitis (HSVE). By using these two assays as well as three markers for blood‐brain barrier, leakage can be easily excluded. A total of 21 sera and 24 cerebrospinal fluid (CSF) samples from 11 patients with HSVE were examined. For seven patients more than one pair of serum and CSF were available. For one patient IgG antibodies began to be detectable in CSF after the sixth day from the onset of the disease. In the other 10 patients the intrathecal synthesis of HSV IgG antibodies was detected later than the sixth day and reached high optical density (OD) values after the 10th day from the onset of disease, at the earliest. In contrast, intrathecal HSV antibody synthesis was not found in specimens taken from 20 patients with acute meningitis who composed our negative control group. The use of a combined indirect ELISA and of an immunoblotting assay on a single dilution of serum and CSF for HSV IgG synthesis in the central nervous system (CNS) allowed the diagnosis of HSVE after the first week of disease.
Expression of urokinase‐type plasminogen activator and plasminogen activator inhibitor in colon disease
Authors: Miseljic Slobodan , Galandiuk Susan , Myers Stephen D. , Wittliff James L.
Published: March 2007

Surveillance colonoscopy and biopsy are inaccurate methods of predicting the likelihood of ulcerative colitis patients to develop colon carcinoma. We examined uPA and PAI‐1 as potential markers for assessing these patients and those with familial polyposis who are at risk of developing colon cancer. For comparison, biopsies of normal colon and Crohn's disease were evaluated. We examined 77 colonic mucosa specimens taken from patients undergoing elective resection for benign and malignant colonic disease. uPA and PAI‐1 were measured using a monoclonal antibody‐based ELISA kit (American Diagnostica, Greenwich, CT) and expressed as ng/mg extract protein. Intra‐ and interassay controls of uPA gave CV = 3–4% and CV = 8–9%, respectively, while those for PAI‐1 were 6–7% and 10–11%, respectively. The Mann‐Whitney test showed that both uPA and PAI‐1 expression were significantly higher in colon cancer, chronic ulcerative colitis, and Crohn's disease than in normal colon. uPA in familial polyposis samples was similar to that of normal colon, while PAI‐1 was much lower than in normal colon. Neither patient age nor sex appeared to influence the expression of these potential markers in any tissue. The pattern of uPA and PAI‐1 expression in normal, benign and malignant colon suggests these proteins deserve further consideration as markers for assessing colon carcinoma risk.
Expression pattern of the water channel aquaporin‐4 in human gliomas is associated with blood–brain barrier disturbance but not with patient survival
Authors: Warth Arne , Simon Perikles , Capper David , Goeppert Benjamin , Tabatabai Ghazaleh , Herzog Hajo , Dietz Klaus , Stubenvoll Florian , Ajaaj Rami , Becker Ralf , Weller Michael , Meyermann Richard , Wolburg Hartwig , Mittelbronn Michel
Published: March 2007

Aquaporin‐4 (AQP4), the most prominent CNS water channel, is restricted to the glia limitans and astrocytic endfeet. We previously showed the loss of spatial AQP4 expression in glioblastomas and a redistribution across the cell surface. However, opposing AQP4 functions have been described: protective in vasogenic but detrimental in cytotoxic brain edema. Thus, specific AQP4 induction to prevent or reduce vasogenic edema is suggested. To elucidate the AQP4 role in brain tumors, we investigated 189 WHO grade I–IV gliomas by immunohistochemistry and the prognostic significance for patients' survival. In gliomas, a remarkable de novo AQP4 redistribution was observed in comparison with normal CNS tissue. Surprisingly, the highest membraneous staining levels were seen in pilocytic astrocytomas WHO grade I and grade IV glioblastomas, both significantly higher than in WHO grade II astrocytomas. AQP4 up‐regulation was associated with brain edema formation; however, no association between survival and WHO grade‐dependent AQP4 expression was seen. Hence, AQP4 redistribution may go along with other tumor properties, such as vascular proliferation and resulting blood–brain barrier disturbance, features usually prominent in pilocytic astrocytomas WHO I and glioblastomas WHO grade IV. In summary, our findings question the protective role of AQP4 in vasogenic brain edema. Although AQP4 was associated with brain edema formation, one has to question the suitability of AQP4 induction as a promising approach in vasogenic brain edema prevention and treatment. In addition, our results provide unexpectedly high AQP4 levels in pilocytic astrocytomas and present AQP4 as tumor progression marker in WHO grade II–IV astrocytomas. © 2007 Wiley‐Liss, Inc.
Transplantation of post‐mitotic human neuroteratocarcinoma‐overexpressing Nurr1 cells provides therapeutic benefits in experimental stroke: In vitro evidence of expedited neuronal differentiation and GDNF secretion
Authors: Hara Koichi , Matsukawa Noriyuki , Yasuhara Takao , Xu Lin , Yu Guolong , Maki Mina , Kawase Takeshi , Hess David C. , Kim Seung U. , Borlongan Cesar V.
Published: March 2007

Nurr1 has been implicated as a transcription factor mediating the endogenous neuroprotective mechanism against stroke. We examined the in vivo and in vitro properties of a new human embryonic carcinoma Ntera‐2 cell line carrying the human Nurr1 gene (NT2N.Nurr1). Adult Sprague‐Dawley rats underwent experimental stroke initially and 14 days later were assigned randomly to receive stereotaxic transplantation of NT2N.Nurr1 cells or infusion of vehicle into their ischemic striatum. Transplantation of NT2N.Nurr1 cells promoted significant attenuation of behavioral impairments over a 56‐day period after stroke, characterized by decreased hyperactivity, biased swing activity, and neurologic deficits, as well as significant reduction in ischemic striatal cell loss compared to vehicle‐infused stroke animals. Transplanted NT2N.Nurr1 cells survived and expressed neuronal phenotypic markers in the ischemic striatum. In vitro results showed that cultured NT2.Nurr1 cells were already negative for nestin even before retinoic acid treatment, despite strong nestin immunoreactivity in NT2 cells. This indicates Nurr1 triggered a rapid commitment of NT2 cells into a neuronal lineage. Indeed, NT2.Nurr1 cells, at 4 weeks into RA treatment, displayed more abundant tyrosine hydroxylase positive cells than NT2 cells. Parallel ELISA studies showed further that cultured NT2N.Nurr1, but not NT2N cells, secreted glial cell derived neurotrophic factor. The present study shows efficacy of NT2N.Nurr1 cell grafts in ischemic stroke, with in vitro evidence suggesting the cells' excellent neuronal differentiation capability and ability to secrete GDNF as likely mechanisms mediating the observed therapeutic benefits. © 2007 Wiley‐Liss, Inc.
Neuritin (cpg15) enhances the differentiating effect of NGF on neuronal PC12 cells
Authors: Cappelletti Graziella , Galbiati Mariarita , Ronchi Cristina , Maggioni Maria Grazia , Onesto Elisa , Poletti Angelo
Published: March 2007

Neuritin is a small, highly conserved GPI‐anchored protein involved in neurite outgrowth. We have analyzed the involvement of neuritin in NGF‐induced differentiation of PC12 cells by investigating the time‐course of neuritin expression, the effects of its overexpression or silencing, and the possible mechanisms of its regulation and action. Real‐time PCR analysis has shown that neuritin gene is upregulated by NGF in PC12 cells hours before neurite outgrowth becomes appreciable. PC12 cells transfected with a plasmid expressing neuritin display a significant increase in the response to NGF: 1) in the levels of SMI312 positive phosphorylated neurofilament proteins (markers for axonal processes) and tyrosine hydroxylase; 2) in the percentage of cells bearing neurites; as well as 3) in the average length of neurites when compared to control cells. On the contrary, neuritin silencing significantly reduces neurite outgrowth. These data suggest that neuritin is a modulator of NGF‐induced neurite extension in PC12 cells. We also showed that neuritin potentiated the NGF‐induced differentiation of PC12 cells without affecting TrkA or EGF receptor mRNAs expression. Moreover, the S‐methylisothiourea (MIU), a potent inhibitor of inducible nitric oxide synthases, partially counteracts the NGF‐mediated neuritin induction. These data suggest that NGF regulates neuritin expression in PC12 cells via the signaling pathway triggered by NO. This study reports the first evidence that neuritin plays a role in modulating neurite outgrowth during the progression of NGF‐induced differentiation of PC12 cells. PC12 cells could be considered a valuable model to unravel the mechanism of action of neuritin on neurite outgrowth. © 2007 Wiley‐Liss, Inc.
Determination of the protein content of wheat and barley by direct alkaline distillation
Authors: Ronalds John A.
Published: March 2007

An improved alkaline distillation method is described for determining the protein content of wheat or barley. A complete estimation can be performed in about 15 minutes. The sample is distilled with a solution containing 10 % sodium hydroxide, barium chloride and antifoam and a specified volume of distillate collected and titrated with acid. Equations are given for calculating the protein content of the sample from the titration value. Different calibration equations are necessary for bread wheats, durum wheats and barley. Excellent agreement with the Kjeldahl method was obtained for analyses of the three grain types with correlation coefficients varying from 0.993 to 0.998 and standard errors ranging from 0.17 to 0.19% protein.
Effects of supranutritional dietary vitamin E levels on subcellular deposition of α‐tocopherol in the muscle and on pork quality
Authors: Asghar Ali , Gray J Ian , Booren Alden M. , Gomaa E A , Abouzied Mohamed M , Miller Elwyn R , Buckley D Joseph
Published: March 2007

The influence of three levels of vitamin E in the diet of pigs on the subcellular deposition of α‐tocopherol in the muscle and on selected quality characteristics of pork meat (oxidative stability of lipids, colour, drip loss, microbial growth) was studied.

The content of α‐tocopherol in adipose tissue and L. dorsi muscle as well as in mitochondrial and microsomal fractions of the muscle significantly increased (P < 0.05) with increasing levels of dietary vitamin E. The differences in the concentrations of α‐tocopherol in the subcellular fractions were evident in the enhanced stability of the membranes when exposed to metmyoglobin/hydrogen peroxide.

The beneficial effect of dietary vitamin E on the oxidative stability of pork lipids during the storage of pork chops and ground pork was also demonstrated. Even though lipid oxidation increased in all cases during storage, the pork products from the pigs receiving the highest level of vitamin E (200 IU kg−1 feed) exhibited the smallest increase in thiobarbituric acid reactive substances. In addition, increased colour stability and decreased drip loss were observed on keeping pork chops, which had been previously frozen for three months, at 4°C under fluoresent light for 10 days. The possible effect of α‐tocopherol on membrane fluidity in this context is discussed.
Influence of must composition on phenolic oxidation kinetics
Authors: Rigaud Jacques , Cheynier Véronique , Souquet Jean‐Marc , Moutounet Michel
Published: March 2007

White grape musts with added sulphur dioxide, ascorbic acid, glutathione, copper or flavanols were prepared and oxidised using known amounts of oxygen. Sulphur dioxide and ascorbic acid delayed caffeoyltartaric acid oxidation, 2‐S‐glutathionyl caffeoyltartaric acid (GRP) formation and browning development. The glutathione content determined the quantity of GRP formed; browning was prevented when all the caffeoyltartaric acid quinones generated by enzymic oxidation could be trapped as GRP. Cupric ions, which lower the concentration of available glutathione, enhanced must browning. Coupled oxidation of flavanols by the caffeoyltartaric acid quinones also led to increased discoloration.
Rumen microbial degradation of allyl cyanide as a possible explanation for the tolerance of sheep to brassica‐derived glucosinolates
Authors: Duncan Alan J , Milne John A
Published: March 2007

Previous studies have shown that sheep are relatively tolerant to the prop‐2‐enyl glucosinolate breakdown product, allyl cyanide (ACN). In order to examine the extent to which this is due to degradation of the toxin in the rumen, the in‐vitro stability of ACN in sheep rumen fluid was determined. Gas chromatography of the volatiles in rumen fluid sequentially sampled following addition of ACN revealed a steady reduction in ACN concentration in the rumen fluid of cabbage‐fed sheep (1.0 μmol ml−1 to 0.36 μmol ml−1 in 100 min) but not in that from sheep fed dried grass (1.0 μmol ml−1 unchanged after 160 min). In a further experiment the time taken to develop an adapted rumen microbial population was determined by switching sheep abruptly from a grass to a cabbage diet and measuring the ‘ACN‐degrading potential’ of rumen fluid over 29 days. Adaptation was rapid (< 6 days) and could confer tolerance to the effects of ACN to sheep consuming forage brassica crops.
Heat stability of trypsin inhibitors in tropical root crops and rice and its significance for nutrition
Authors: Bradbury J Howard , Hammer Brendon C , Sugani Inge
Published: March 2007

The heat stability of trypsin inhibitors from rice (Oryza sativa L), sweet potato (Ipomoea batatas (L) Lam), taro (Colocasia esculenta var esculenta (L) Schott), giant taro (Alocasia macrorrhiza (L) G Don) and giant swamp taro (Cyrtosperma chamissonis (Schott) Merr) was studied. Rice trypsin inhibitor (TI) occurred only in the germ, so it is absent from white rice. Taro, giant taro and giant swamp taro (but not rice or sweet potato) showed an initial large increase (one‐ to tenfold) in the amount of TI present on heating at about 80°C. The heat release mechanism, which was accompanied by a change to a rubbery texture in the food, amounted to a breakdown of the cellular structure making TI available for extraction. Such heat release of TI would cause partially cooked food to be deleterious for human consumption. Brown rice, sweet potato, taro and giant swamp taro contained moderate amounts of TI, which was fully inactivated when the boiled or baked food was soft enough to eat. Giant taro, however, contained 10–100 times as much TI (which in this case also inhibited chymotrypsin) as the other foods, and a boiling time of at least 30 min was needed for its inactivation.
Protein content variation in a pure line faba bean (Vicia faba) collection
Authors: El‐Sherbeeny Mohammed H , Robertson Larry D
Published: March 2007

A set of 840 pure line faba beans (Vicia faba L) derived by a process of cyclic single plant selection from 1979 to 1985 was used for this study. Mean content of protein was 240 g kg−1 with a standard deviation of 18 g kg−1 and a range of 180 to 310 g kg−1. For eight subregional groups (Nile Valley, West Asia, Northern Europe, Southern Europe, Eastern Europe, North Africa, Ethiopia and the Indian subcontinent) and for a large and small grouping, the means and ranges of protein content were similar. There were no significant correlations between protein content and seed yield or its components. This resulted in a strong correlation (significant at P < 0.01) between seed yield and protein yield (r = 0‐98), supporting the hypothesis that the optimal way to increase sustainable protein yield per hectare is to increase and stabilise seed yield.
Storage of fresh and ensiled forages by freezing affects fibre and crude protein fractions
Authors: Kohn Richard A , Allen Michael S
Published: March 2007

The effect of freezing on fibre and crude protein fractions of forages was determined. Fresh and ensiled lucerne and fresh bromegrass were processed immediately after collection or stored at −25°C for 1 day or 1, 6 or 12 months before drying at 55°C. Samples were frozen quickly by submersion in liquid nitrogen or slowly at −25°C. Samples which were not frozen were processed immediately or after 1 h delay at room temperature. All treatments were replicated (n = 3). Samples were analysed for crude protein (CP), trichloroacetic acid soluble CP (TSCP), phosphate buffer soluble CP (BSCP), neutral detergent insoluble CP (NDCP), acid detergent insoluble CP (ADCP), neutral detergent fibre (NDF), acid detergent fibre (ADF), acid detergent sulphuric acid lignin (lignin) and ash. Freezing decreased BSCP and increased NDCP by more than 40% for bromegrass. Freezing also changed NDF, ADF, lignin, ash, CP and ADCP in different ways depending on forage type and length of time frozen. No significant effects were observed for method of freezing or a 1‐h delay in processing. An additional experiment showed that freeze‐drying resulted in less insoluble protein than ovendrying. Prior freezing of forages appeared inconsistently to change the extent of gaseous loss during drying, and resulted in precipitation of protein. These changes also affected fibre estimates. Fresh forages should not be frozen and thawed before analysis of protein or fibre fractions.
Effect of chloride salts on protein extraction and interfacial protein film formation in meat batters
Authors: Gordon Andre , Barbut Shai
Published: March 2007

Six comminuted chicken breast meat mixes and six meat batters were made with isoionic NaCl (25 g kg−1), MgCl2, CaCl2, KCl, LiCl (IS = 0.43) and 15 g kg−1 NaCl (IS = 0.26). The quantity and type of proteins extracted and used for interfacial protein film (IPF) formation was determined and related to batter stability. The monovalent salts produced IPF which differed in individual protein content between salts but which all contained significantly larger amounts of protein (P < 0.01) than those using divalent salts. MgCl2 extracted more protein than CaCl2 and produced a different protein profile in the IPF. However, MgCl2 formed unstable raw batters whereas CaCl2 did not. In addition, a simple, rapid method for extracting and quantifying proteins from meat batters was developed to assist in direct determination of the actual soluble protein uptake by the fat phase during comminution.
Control of wheat α‐amylase using inhibitors from cereals
Authors: Henry R. J. , Battershell V G , Brennan P S , Oono K
Published: March 2007

A survey of 46 varieties of cereals and related species (including 27 different species from the Poaceae) indicated the presence of a strong inhibitor of wheat α‐amylase in all seven Hordeum species tested. Rye contained a lower level of inhibitor activity, but the other species contained insignificant amounts of wheat α‐amylase inhibitor activity. The partially purified barley inhibitor was most effective in inhibiting wheat α‐amylase activity at high pH. The addition of chromosome 2 of barley to wheat (Chinese Spring addition line 2H) resulted in an apparent increase in the molecular weight of the α‐amylase produced during germination. This was probably due to the formation of a complex between the inhibitor encoded by the asi gene on chromosome 2 of barley and wheat α‐amylase 2. Breeding of wheat with the barley inhibitor gene may reduce the impact of the high α‐amylase levels that result from pre‐harvest sprouting in wheat.
Low trans hydrogenation of edible oils
Authors: Beers Annemarie E.W. Beers
Published: March 2007

Hydrogenation of edible oils is an important process in the food industry to produce fats and oils with desirable melting properties and an improved shelf life. However, beside the desired hydrogenation reaction trans fatty acids are formed as well. As several studies indicate a negative health effect of trans fatty acids, consumer demands will urge the food producers to lower the content of trans fatty acids in their products. This article describes the option to reduce the trans levels in the hydrogenation of an edible oil by changing process conditions and by applying alternative low trans heterogeneous catalysts.
The response to antioxidants in base oils of different degrees of refining
Authors: Wiklund Per
Published: March 2007

Base oils with different compositions and different degrees of refining have been subjected to three types of oxidation stability tests in the presence of antioxidants. A higher degree of refining of base oils depletes the natural antioxidants present in oils of lower degree of refining. However, the results of the study show that oxidation stability of highly refined oils, after addition of suitable antioxidant packages, by far outperforms that of the oils of lower degree of refining. This is true regardless of the dominating hydrocarbon type in the oil, or its feedstock origin. The results also show that in general, the viscosity index of a base oil, or any other simple physical property, has no direct correlation to oxidation stability or response to antioxidants.

To analyse the response to antioxidants, a concept called oxidation stability response is introduced. A novel method for determining the oxidation onset temperature in differential scanning calorimetry measurements by inverse derivatives is introduced. Copyright © 2007 John Wiley & Sons, Ltd.
Improved techniques for kidney transplantation in mice
Authors: Zhang Zheng , Schlachta Christopher , Duff John , Stiller Calvin , Grant David , Zhong Robert
Published: March 2007

Improved microsurgical techniques for kidney transplantation in the mouse are described. Left renal transplantation is performed with end‐to‐side anastomoses of the donor renal vein to the inferior vena cava and the donor aortic cuff to the aorta. Urinary tract reconstruction is accomplished by suturing the donor ureter with a bladder patch to the recipient bladder. With these modifications, it is possible to achieve success rates as high as 90%. This stable and reproducible model provides a useful tool to study the immunological mechanisms of kidney allograft rejection at the molecular level. © 1995 Wiley‐Liss, Inc.
Hemodynamic study of different angled end‐to‐side anastomoses
Authors: Zhang Ling , Moskovitz Martin , Piscatelli Sandra , Longaker Michael T. , Siebert John W.
Published: March 2007

This study assessed the postoperative microvascular hemodynamics in end‐to‐side arterial anastomoses of varied angles in order to determine whether the angle of the anastomosis affects arterial flow. Thirty male rats were divided into three groups of ten. Carotid to carotid end‐to‐side anastomoses were performed with anastomotic angles of 45, 90, and 135°. Postoperative flow was assessed using high frequency pulsed Doppler ultrasound (HFPDU) at 30 and 120 min postoperatively. Measurements at 30 min demonstrated significant differences in flow, with the 45° grafted vessels utilizing 56.5% of total carotid flow, and the 90° and 135° anastomoses appropriating 46.5% and 43.2% of flow, respectively (comparing 45° to both 90°, P < 0.05, and 135°, P<0.002). The change from baseline in 45° and 90° groups dissipated over a 2 hr postoperative period, but flow in the grafted vessels in the 135° group continued well below 50% at 39.9% (comparing to both 45° and 90°, P<0.001), thereby displaying a significant difference in the postoperative arterial flow of varied angled microanastomoses. © 1995 Wiley‐Liss, Inc.
Vein conduits for repair of nerves with a prolonged gap or in unfavorable conditions: An analysis of three failed cases
Authors: Tang Jin‐Bo , Shi De , Zhou Hua
Published: March 2007

Clinical failure of vein conduits for repair of four nerves in three cases is reported. Two digital nerves with gaps of 5.0 cm and 5.8 cm, respectively, and two median nerves with gaps of 4.0 cm and 5.0 cm were repaired with vein conduits. The digital nerves were repaired secondarily with insertion of nerve tissue slices. The median nerve lacerations were associated with compound soft tissue injuries and were repaired primarily by interfascicular vein conduits. There was no detectable recovery of sensibility in autonomous areas of these nerves and no sign of recovery of the innervated muscles during follow‐up. Re‐exploration revealed that the vein conduits used for repair of the median nerves were constricted by surrounding scar tissue and axon regeneration was precluded. The critical length for nerve regeneration in human beings and wound conditions unfavorable to nerve regeneration are discussed. This report suggests that vein conduits are not indicated in nerve gaps over 5.0 cm or in primary repair of nerves with compound injuries. © 1995 Wiley‐Liss, Inc.
Can varying flow velocity across an arterial anastomosis prevent thromboembolic injury?
Authors: Barker John H. , Andresen Dorthe M. , Anderson Gary L. , Schuschke Dale , Gu Jin‐Mei , Gupta Subhas , Hjortdal Vibeke E. , Derr John W. , Banis Joseph C. , Acland Robert D.
Published: March 2007

In this study, simulated “poor” repairs applied to transverse incisions in the iliac arteries of 40 rats were the basis for comparing the effect of variations in blood flow on throm‐boembolism. Using vital microscopy and digital image processing, we performed 2 experiments. In the first experiment (n = 20), the reduction of post‐repair blood flow by approximately 50% resulted in an 83% reduction in the total number of emboli appearing in the microcirculation of the cremaster muscle distal to the repair. In the second experiment (n = 20), the same reduction in blood flow typically resulted in larger repair‐site thrombi which required significantly more time to grow to their maximum size. We conclude that reducing pedicle artery blood flow to approximately half in our rat model during reperfusion can protect the downstream micro‐circulation from embolic injury without increasing the incidence of thrombotic occlusion. © 1995 Wiley‐Liss, Inc.
Free flap failures
Authors: Suominen Sinikka , Asko‐Seljavaara Sirpa
Published: March 2007

A retrospective analysis of 75 consecutive free flap patients, operated on during 1989–1990, was performed to find out more about factors associated with free flap failure or immediate vascular complications. The overall failure rate was 9.3% (7/75) and the immediate vascular complication rate 22.7% (17/75). Sixteen patients required explorative surgery during the first postoperative day. The results were statistically analysed to find factors promoting either failure or vascular complications. Pre‐operative infection of the recipient site or prolonged operation time correlated with flap failure. The use of a vein graft or long per‐operative ischaemia correlated with immediate vascular complications. It is interesting that many factors often blamed for failure (age, body mass, history of cardiovascular disease, smoking, or previous irradiation of the recipient site) were not significant in this study. © 1995 Wiley‐Liss, Inc.
Forearm arterial loop as an expedient source for inflow to upper extremity free flaps
Authors: Hallock Geoffrey G.
Published: March 2007

Challenging wounds of the upper extremity requiring free tissue transfers are relatively infrequent. As a direct consequence, methods to access recipient vessels or acceptable alternatives may not be as familiar as in the lower extremity. In rare circumstances, creation of an arterial loop by elevation of a portion of either major forearm artery provides an easily accessible, pristine source of inflow for a planned free flap matching vessels of comparable caliber. Such a maneuver may also be important to ensure preservation of optimal hand function by avoiding any dissection in the vicinity of essential musculotendinous and neurovascular structures that might reside within or adjacent to the zone of injury. © 1995 Wiley‐Liss, Inc.
Mandibular reconstruction using the double barrel fibular graft
Authors: Horiuchi Katsuhiro , Hattori Akinobu , Inada Ikuhisa , Kamibayashi Toyohiko , Sugimura Masahito , Yajima Hiroshi , Tamai Susumu
Published: March 2007

Five patients underwent mandibular reconstruction using the double barrel fibular graft from 1989 to 1994. Bony defects ranged from 7 to 14 cm. In three patients, two skin flaps were taken with the fibular graft for composite reconstruction. In order to overcome the main disadvantage of the fibular graft, i.e., small circumference of the bone, a harvested fibula was osteotomized into several portions, folded into two parallel lengths, and fixed along the inferior border of the mandible and the alveolar ridge. The double barrel fibular graft provided more than 4‐cm alveolar height without damaging bone viability. In Orientals, a fibula is approximately 1.5 cm thick, and using a single fibular strut for mandibular reconstruction may result in subsequent difficulty in wearing conventional dentures or osseointegrated implants. All patients acquired good mandibular contour and enough thickness of the alveolar ridge, and could wear a conventional denture and eat a solid diet. This procedure seems to be superior to the iliac bone graft for major mandibular reconstruction because of its length, the possibility of three‐dimensional composite reconstruction, increased bone thickness, and minimal donor‐site morbidity. © 1995 Wiley‐Liss, Inc.
Effects of nerve growth factor on crushed sciatic nerve regeneration in rats
Authors: Chen Zhong‐Wei , Wang Min‐Sheng
Published: March 2007

The effects of nerve growth factor (NGF) on crushed sciatic nerve regeneration were studied in 30 rats, with 60 bilateral nerves. The nerves were crushed at a site 6 mm distal to the sciatic notch by the standard technique and 3 mm wide crush injuries were created. Then 2.1 μl of normal saline in the control groups and an equal volume of NGF solution (containing 1 μg of NGF) in the NGF‐treated groups was injected into the crush sites and followed for 12, 28, and 56 days, respectively. At the end of the observation, electrophysiological evaluation was carried out; then samples 10 mm distal to the crush site were removed and prepared for histological and morphometric studies. Evoked muscle action potential (MAP) was recorded in 50% of the NGF‐treated group at 12 days but not in the control group; the difference was statistically significant (P<0.05). The motor nerve conduction velocity (MNCV) was increased in NGF‐treated groups compared with control groups at 28 and 56 days (P<0.05). Morphometrically, significantly more regenerated myelinated fibers (RMFs) were seen at 12 days, and larger diameter RMFs were found at 12, 28, and 56 days in NGF‐treated groups than in control groups. These results indicate that topically applied NGF stimulates nerve regeneration and promotes function recovery in crushed rat sciatic nerves. © 1995 Wiley‐Liss, Inc.
Delayed sensory reeducation after toe‐to‐hand transfer
Authors: Wei Fu‐Chan , Ma Hae‐Shya
Published: March 2007

The effect of a delayed program of sensory rehabilitation was evaluated in 13 patients with a total of 22 toe‐to‐hand transfers. Each patient was begun on a protocol of sensory reeducation at a mean of 38 months (range 13–98 months) after the transfer. Home rehabilitation, employing a Disk‐Criminator (TM) program, was monitored at monthly intervals by the therapist for a mean of 3.3 months, at which time a final evaluation of sensibility was done. Each of the 22 toe transfers improved by an average of 7 mm in static and 6 mm in moving two‐point discrimination. The improvement in two‐point discrimination following delayed implementation of sensory reeducation was statistically significant at the P<0.0001 level. © 1995 Wiley‐Liss, Inc.
Convergence of coercive approximations for strictly monotone quasistatic models in inelastic deformation theory
Authors: Chełmiński Krzysztof , Gwiazda Piotr
Published: March 2007

This article studies coercive approximation procedures in the infinitesimal inelastic deformation theory. For quasistatic, strictly monotone, viscoplastic models using the energy method and the Young measures approach a convergence theorem in generalized Orlicz spaces is proved. The main step in the proof is a characterization of the weak limit of non‐linear terms by the convergence in measure. Copyright © 2007 John Wiley & Sons, Ltd.
Novel oxidative in vitro metabolites of the mycotoxins alternariol and alternariol methyl ether
Authors: Pfeiffer Erika , Schebb Nils H. , Podlech Joachim , Metzler Manfred
Published: March 2007

The Alternaria toxins alternariol (AOH; 3,7,9‐trihydroxy‐1‐methyl‐6H‐benzo[c]chromen‐6‐one) and alternariol methyl ether (AME, 3,7‐dihydroxy‐9‐methoxy‐1‐methyl‐6H‐benzo[c]chromen‐6‐one) are common contaminants of food and feed, but their oxidative metabolism in mammals is as yet unknown. We have therefore incubated AME and AOH with microsomes from rat, human, and porcine liver and analyzed the microsomal metabolites with HPLC and GC‐MS/MS. Seven oxidative metabolites of AME and five of AOH were detected. Their chemical structures were derived from their mass spectra using deuterated trimethylsilyl (TMS) derivatives, and from the information obtained from enzymatic methylation. Several of the metabolites were identified by comparison with synthetic reference compounds. AME as well as AOH were monohydroxylated at each of the four possible aromatic carbon atoms and also at the methyl group. In addition, AME was demethylated to AOH and dihydroxylated to a small extent. As the four metabolites arising through aromatic hydroxylation of AME and AOH are either catechols or hydroquinones, the oxidative metabolism of these mycotoxins may be of toxicological significance.
Natural‐abundance 13C NMR of brain
Authors: Bárány Michael , Arús Carlos , Chang Yen‐Chung
Published: March 2007

In natural‐abundance 13C NMR spectrum of excised rat brain 55 resonances were resolved. The chemical shifts of most of the resonances were well correlated with those of pure brain metabolites, determined under identical experimental conditions. These resonances were also found in the cytosol fraction and perchloric acid extract of the brain. Some brain resonances were not observed in the perchloric acid extract but only in the microsomes or cytosol.