bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

AbstractIt 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

AbstractWe 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

AbstractA 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

AbstractA 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

AbstractSurveillance 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

AbstractAquaporin‐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

AbstractNurr1 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

AbstractNeuritin 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

AbstractAn 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

AbstractThe 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

AbstractWhite 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

AbstractPrevious 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

AbstractThe 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

AbstractA 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

AbstractThe 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

AbstractSix 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

AbstractA 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.