bluuebunny/crossref_metadata_2025_split huggingface dataset:
Application of checkerboard immunoblotting (CBIB) to the detection of anti‐viral IgG in human serum
Authors: Craig Wendy Y. , Poulin Sue E. , Dorsett Preston H. , Ledue Thomas B. , Ritchie Robert F. 
Published: March 2007

AbstractIn the present study, we have begun to investigate the possibility of using checkerboard immunoblotting (CBIB) as a semi‐quantitative screening tool for detecting human serum IgG against specific viral antigens. The viral antigens studied were Epstein‐Barr, herpes simplex I and II, cytomegalovirus, varicella zoster, rubella, rubeola, and mumps. Western immunoblotting experiments using these partially purified preparations demonstrated that there were apparently no interactions between IgG from non‐immune sera and the respective viral antigen preparations. The CBIB assay was evaluated using sera of known positive or negative immune status for the viral antigens. There was excellent agreement between the results of CBIB and the results of alternative methods for evaluating immune status; all discrepancies (1/18 sera for mumps, 3/18 sera for rubeola, and 1/28 sera for rubella) involved sera with borderline results, either by CBIB or by the alternative method. Therefore, although further work is required to define the method in terms of sensitivity and clinical specificity, and to refine positive/negative cutpoint criteria for certain antigen components, our preliminary experience suggests that CBIB has considerable potential in the efficient and inexpensive screening of sera for the presence of IgG against a panel of viral antigens, so as to identify subjects at risk for infection. © 1993 Wiley‐Liss, Inc.
Peroxidase/monoclonal antibody conjugates for the study of hemoglobinopathies
Authors: Moscoso Hugo , Kiefer Charles R. , Shyamala M. , Garver Fred A. 
Published: March 2007

AbstractMonoclonal antibodies (mAbs) to normal human hemoglobins (Hbs) A and F and to variant Hbs C and G‐Philadelphia were conjugated to horseradish peroxidase (HRP) and used in qualitative or quantitative enzymelinked immunosorbent assays (ELISAs). Conjugates with output molar HRP/IgG ratios close to 2.0 had higher avidity for the cognate antigens than those with ratios above or below 2.0. The analytical sensitivities of the conjugates ranged from 0.2 to 4 ng of hemolysate containing the target hemoglobin, and it was not related to the input or the output HRP/IgG ratios. The overall imprecision for the qualitative ELISA was below 8%, and the accuracy for the identification of Hbs C and G‐Philadelphia was 100% as compared with established methods. Quantitative determinations of HbA based upon direct dose‐response curves showed an analytical sensitivity of 1% and an imprecision ≪ 11%. The most significant application of the HbA assay was in the differential diagnosis of hemoglobinopathies associated with partial or total suppression of HbA synthesis. Competitive dose‐response curves for the HRP/ anti‐γ conjugate allowed the quantification of HbF in the clinically significant range of 0.5‐10%, with an imprecision ≪12%. It is concluded that the incorporation of HRP/mAb conjugates into the ELISA technique offers a simpler, more rapid, yet specific alternative for the measurement of hemoglobins. © 1993 Wiley‐Liss, Inc.
Correlation between human sperm hypo‐osmotic swelling test and other semen parameters
Authors: Al‐Mogazy Hassan M. , Al‐Khadra Samih , Kenawy Gaafar , Al‐Zeftawy Ali , Mostafa Taymour , Attia Ahmed , Attia Abdalla M. 
Published: March 2007

AbstractNinety semen samples from male patients were studied. They were classified into 4 different groups; normospermic fertile, oligospermia, asthenospermia, and oligoasthenospermia. In addition to routine semen analysis, the hypo‐osmotic swelling test was done for all these samples, measuring total protein and specific gravity. A significant positive correlation was observed between percent sperm swelling and its motility, its total protein, and specific gravity. Also a significant negative correlation was noticed between percent sperm swelling and its count. Although there were overlapping ranges, different groups showed significant relations of percent sperm swelling among themselves.It is postulated that the sperm quality and not its quantity is the cornerstone of the hypoosmotic test and that the physico‐chemical properties of the sperm influences its swelling. © 1993 Wiley‐Liss, Inc.
Glycoconjugates of Trypanosoma cruzi: A 74 kD antigen of trypomastigotes specifically reacts with lytic anti‐α‐galactosyl antibodies from patients with chronic Chagas disease
Authors: Almeida Igor C. , Krautz Greice M. , Krettli Antoniana U. , Travassos Luiz R. 
Published: March 2007

AbstractProtective, lytic antibodies are believed to be correlated with active Typanosoma cruzi infection. In patients with chronic infection, antibodies lysing trypomastigote forms recognize chiefly α‐galactosyl structures at the parasite surface. The target molecules on cell‐derived trypomastigotes that react with anti‐α‐galactosyl antibodies (anti‐Gal) from patients with chronic Chagas disease were investigated. Glycoconjugates were isolated from trypomastigotes and shown to absorb purified Chagasic (Ch) anti‐Gal effectively as well as lytic antibodies from Ch sera. Active fractions were F2 (74 kD and 95.6 kD) and F3 (120–200 kD). A differential reactivity with antibodies from untreated Ch patients (trypanolytic) and from treated, presumably cured, individuals (not trypanolytic) was evident using F2 and F3 antigenic fractions. No cross‐reactivity with heterologous sera (other infections) was observed. The F2 glycoconjugate (mostly 74 kD) can be used in the diagnosis of active Chagas infection, replacing the quantitative determination of complementmediated lysis. With the present sample of patients' sera and normal human sera, it showed 100% sensitivity and specificity. © 1993 Wiley‐Liss, Inc.
Protein 1: Its purification and application in clinical medicine
Authors: Itoh Yoshihisa , Ishii Shuichi , Okutani Ryuta , Asano Yasushi , Kawai Tadashi 
Published: March 2007

AbstractProtein 1 (P1) is a low‐molecular‐weight protein recently isolated from the urine of patients with chronic renal failure. Its molecular weight is 14 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis and pl 4.7 on isoelectric focusing. We purified this protein, characterized its physicochemical properties, and analyzed its amino acid sequences to show that it is probably identical to human lung Clara cell 10 kDa protein. Its monoclonal antibody was prepared, and a reliable enzyme‐linked immunosorbent assay employing the sandwich method was developed and used to investigate distribution and variation in concentration of P1 in various body fluids under an array of physiologic and pathologic conditions. Clinical studies indicated that, as is the case with other proteins of low molecular weight, the main catabolic site of P1 of plasma origin is the kidney: P1 is filtered by the renal glomeruli and reabsorbed by the renal tubules. Unabsorbed P1 is thus excreted into the urine. This protein is also synthesized in the genital tissues of males, however, from which it is also excreted into the urine. Clinical data obtained in our study of this protein is summarized here, and an attempt is made to determine the potential value of this protein in laboratory medicine. © 1993 Wiley‐Liss, Inc.
Ectopic production of prostate specific antigen by a breast tumor metastatic to the ovary
Authors: Yu H. , Diamandis E. P. , Levesque M. , Sismondi P. , Zola P. , Katsaros D. 
Published: March 2007

AbstractWe have recently reported that about 30% of breast tumors produce prostate specific antigen (PSA). We examined here, 99 primary ovarian cancer tumors and found relatively low levels of PSA in only three tumors. One patient with metastatic ovarian cancer from a primary breast tumor, produced relatively high levels of PSA and responded well to antiestrogen treatment although she was steroid receptor‐negative. Another patient with metastatic ovarian cancer from a primary breast tumor did not produce PSA and did not respond to treatment although she was steroid hormone receptor‐positive. This data describe for the first time, ectopic PSA production by a breast tumor at the ovarian metastatic site and further support the view that PSA is a favourable prognostic indicator in breast cancer. © 1994 Wiley‐Liss, Inc.
Rapid fluorometric assay for mitochondrial proton adenosine triphosphatase activity for assessment of viability of liver graft tissue
Authors: Seya Kazuhiko , Ohkohchi Nobuhiro , Watanabe Nobuyuki , Shibuya Hiroshi , Taguchi Yoshio , Mori Shozo 
Published: March 2007

AbstractWe developed an improved determination method of mitochondrial proton adenosine triphosphatase (ATPase) activity in the liver. The activity was measured fluorometrically with a 3,3′‐dipropylthiodicarbocyanine iodide (diS‐C3(5)), which is excited at 625 nm and emits fluorescence at 670 nm. This dye transmits the electric potential across the inner mitochondrial membrane. The fluorescence intensity of diS‐C3(5) with mitochondria (100 μl, 4–16 mg/ml protein) in a 2 ml potassium buffer (pH 7.4) was regarded as a standard electric potential. After confirming the activity of the mitochondrial electron transport chain by succinic acid (9 μmol), we inhibited the chain by antimycin A (1.25 μg). Fluorescence intensity decreased by adenosine 5′‐triphosphate (ATP) (2 μmol) and oligomycin (25 μg) inhibited this depression. The value of mitochondrial proton ATPase activity was calculated as a percentage of the fluorescence intensity change by ATP per the standard electric potential. The activity of mitochondrial proton ATPase in the normal fresh rat livers was 50.3 ± 2.2%. Good correlation (r2 = 0.807) between two methods for mitochondrial proton ATPase activity, our newly developed method and a conventional colorimetric method, was obtained in the rat livers with various conditions. This method has advantages that the proton ATPase activity can be measured in intact mitochondria, and all procedures can be completed within 40 min. It is suitable for the determination of mitochondrial viability of liver graft in the hepatic resections and transplantations. © 1994 Wiley‐Liss, Inc.
Apolipoprotein e genotyping methods for the clinical laboratory
Authors: Maekawa Baku , Cole Thomas G. , Seip Richard L. , Bylund David 
Published: March 2007

AbstractTo select the best method for detecting apolipoprotein E (apo E) genotypes determined by the three common alleles ϵ2, ϵ3 and ϵ4, we compared the radiolabeled allele‐specific oligonucleotide (ASO) probe assay and the nonisotopic restriction isotyping assay. Leukocytic DNA was extracted from the blood of 93 patients after which the region containing two mutation points coding amino acid residues 112 and 158 was amplified by using the polymerase chain reaction (PCR). Amplified DNA fragments were spotted on nylon membranes, then hybridized for the ASO probe assay. The amplified DNA fragments were also digested with restriction endonuclease HhaI, followed by polyacrylamide gel electrophoresis for the restriction isotyping assay. The apo E genotypes determined by both methods for every specimen studied were in complete agreement. Although the radiolabeled ASO probe method was 10 times more sensitive than restriction isotyping on polyacrylamide gel, the two were comparable in accuracy. Additionally, because it is simpler to perform, is less time consuming, and is less expensive, we conclude that the restriction isotyping assay is the more suitable of these two methods for use in a clinical laboratory.©1995 wiley‐Liss, inc.
Bioassay vs. immunoassay for quantification of interleukin‐6 in biological fluids
Authors: Papadopoulos Nikolaos G. , Dedoussis George V. Z. , Baxevanis Constantin N. , Papamichail Michail 
Published: March 2007

AbstractAs several possible prognostic and therapeutic applications of interleukin‐6 are currently under trial, the available methods for its quantification in biological fluids should be evaluated. In this report, the 7TD1 hybridoma bioassay is compared to an enzyme immunoassay for the determination of interleukin‐6 in serum and plasma of normal subjects and patients with cancer, sepsis, and systemic lupus erythematosus, as well as in malignant pleural effusions and culture supernatants. The results show a good correlation between the two methods in all cases. Mean values of the examined groups were statistically different between the assays only in the case of septic patients. This may be attributed either to the influence of other molecules on the assays or to the nonlinearity of the dose‐response curves. Since immunoassays are easier to perform, it seems that they are more suitable for routine use, the bioassay being preferable in cases where increased sensitivity is required.
Analysis of whole‐blood cyclosporin g by liquid chromatography in renal transplant recipients
Authors: McBride James H. , Kim Steaven S. 
Published: March 2007

AbstractCyclosporin G (CsG) is less nephrotoxic than Cyclosporin A (CsA) and is undergoing clinical trials for use as an immunosuppressive agent after renal transplantation. In this study, CsG was measured by a rapid high‐performance liquid chromatography (HPLC) technique in blood samples (n=107) received from renal transplant recipients. The HPLC assay proved to be analytically suitable in that it was sensitive, linear, and precise and had high recovery (102%). However, interference was observed from some potentially co‐administered drugs such as calcitriol, ferrous sulfate, hydrazaline, and minoxidil. The HPLC assay for CsG correlated well with a FPIA (Abbott TDx), FPIA = 0.964 (HPLC) + 33.59, r = 0.9819, Sy/x = 36.66 for patients receiving a low dose of CsG (5 mg/kg/day) and a high dose (10 mg/kg/day). Furthermore, the HPLC technique was capable of measuring predictable CsG concentrations when the drug was tapered to lower doses at various stages of the 16 week clinical trial. The HPLC for CsG has the further advantage that the same system and mobile phase can be used to measure CsA while using CsC as the interval standard.
Significance of aneuploidy in non‐hodgkin lymphomas
Authors: Vučković J. , Forenpoher G. , Marušić M. , Užarević B. , Sabioncello A. , Štula N. , Dubravčć M. 
Published: March 2007

AbstractThe incidence of aneuploidy was determined by flow cytometric analysis in paraffin‐embedded tumor samples of 125 patients with non‐Hodgkin lymphoma (NHL). There were 48 low‐grade (LG) and 77 high‐grade (HG) tumors. Aneuploidy was found in 34 (27%) NHL, 15 (31%) LG, and 19 (25%) HG tumors. The analysis of patient survival showed a significantly better survival of patients with LG than HG NHL, but the presence of aneuploidy did not influence the survival on the level of all patients, patients with LG or patients with HG NHL. In patients with LG NHL a tendency of diploid vs. aneuploid patients to survive longer was observed, but only at P = 0.056 significance level.
Immunoreactive prostate‐specific antigen in lung tumors
Authors: Levesque Michael , Yu He , D'Costa Mario , Tadross Latif , Diamandis Eleftherios P. 
Published: March 2007

AbstractProstate‐specific antigen (PSA) is a glycoprotein produced by the epithelial cells of the prostate. PSA is currently used clinically to diagnose and monitor prostate carcinoma. In previous work we have demonstrated that 30% of breast tumors and, more rarely other tumors, contain significant amounts of PSA. PSA appears to be a favorable prognostic indicator in breast cancer. Here, using a sensitive assay, we demonstrated for the first time that lung adenocarcinomas and squamous cell carcinomas also contain PSA. PSA in lung tumor extracts was present mainly in its 33 KDa form (free PSA), at levels measurable by commercial methods. The presence of PSA was associated more closely with male patients and adenocarcinomas. The physiological role of PSA in lung tissue and the prognostic significance of PSA in lung cancer remain to be determined. These and our previous data as well as reports by other groups support the view that PSA is a ubiquitous biochemical marker of steroid hormone action.
Simultaneous age‐related depolarization of mitochondrial membrane potential and increased mitochondrial reactive oxygen species production correlate with age‐related glutamate excitotoxicity in rat hippocampal neurons
Authors: Parihar Mordhwaj S. , Brewer Gregory J. 
Published: March 2007

AbstractMitochondria are implicated in glutamate excitotoxicity by causing bioenergetic collapse, loss of Ca2+homeostasis, and generation of reactive oxygen species (ROS), all of which become increasingly important clinically with age. Little is known about how aging affects the relative importance of mitochondrial membrane potential (ΔΨm) and ROS production. To determine aging affects on ΔΨmand ROS production in individual somal and axonal/dendritic mitochondria, we compared ROS production while simultaneously monitoring ΔΨmbefore and after glutamate treatment of live neurons from embryonic (day 18), middle‐aged (9–12 months), and old (24 months) rats. At rest, old neuronal mitochondria 1) showed a higher rate of ROS production that was particularly strong in axonal/dendritic mitochondria relative to that in middle‐age neurons, 2) were more depolarized in comparison with neurons of other ages, and 3) showed no differences in ROS or ΔΨmas a function of distance from the nucleus. All ΔΨmgrouped into three classes of high (less than –120 mV), medium (–85 to –120 mV), and low (greater than –85 mV) polarization that shifted toward the lower classes with age at rest. Glutamate exposure dramatically depolarized the ΔΨmin parallel with greatly increased ROS production, with a surprising absence of an effect of age or distance from the nucleus on these mitochondrial parameters. These data suggest that old neurons are more susceptible to glutamate excitotoxicity because of an insidious depolarization of ΔΨmand rate of ROS generation at rest that lead to catastrophic failure of phosphorylative and reductive energy supplies under stress. © 2007 Wiley‐Liss, Inc.
Comparison of methods for the determination of available lysine value in animal protein concentrations
Authors: Hussein Laila A. 
Published: March 2007

AbstractA comparison was made between Carpenter's, Erbersdobler's and Rao's methods for the evaluation of available lysine in nine different protein concentrates. The use of methanol is not recommended as a solvent for the FDNB reagent. Measurement at 360 nm gives erroneously higher results. The separation of εDNP‐lysine was best on column chromatography.
The fatty acid composition of the main phospholipid fractions of the rumen and abomasum tissues of foetal and adult sheep
Authors: Body Denis R. , Shorland F. Brian 
Published: March 2007

AbstractThe fatty acid composition has been determined on phosphatidyl choline, phos‐phatidyl ethanolamine and sphingomyelin fractions earlier isolated from the rumen and abomasum tissues of foetal and of adult Romney sheep.The major proportion of polyunsaturated fatty acids was found in the phosphatidyl ethanolamine (17 to 43%) fraction and this was reduced in the phosphatidyl choline (7 to 25%) and sphingomyelin (1 to 4%) fractions. These features are in keeping with the results for mammalian tissues generally. The phosphatidyl ethanolamine fractions were further characterised by the low content of palmitic acid (<8%) compared with 25 to 30 % in the phosphatidyl choline fractions and 29 to 52% in the sphingomyelin fractions and by the occurrence of cyclopropane fatty acids. Consistent with the findings of other workers on mammalian tissues, the sphingomyelin fractions contained a relatively high content (16 to 27%) of higher w‐saturated fatty acids including 22:0,23:0,24:0 and 25:0 and of tetracos‐14‐enoic (24:1 ω9) acid (5 to 16%). The total amounts of acids above C20 tended to vary inversely with the levels of palmitic acid whereas the levels of stearic acid were relatively constant at 13 to 17%.Changes in fatty acid composition with age were generally not marked but the tissues of the foetus were distinguished from those of the foetus were distinguished from those of the adult by their substantial amount of eicosa‐5,8,11‐trienoic (20:3 ω9) acid together with relatively low contents of linoleic (18:2 ω6) and linolenic (18:3 ω3) acids and to a leser extent by reduced level of acids of the ω3 series. This was particularly reflected by the ratios of ω6/ω3 C20 + C22 acids in the phosphatidyl ethanolamine fractions, the valucs for the foetal rumen and abomasum tissues being 1.03 and 1.07, respectively, compared with corresponding values of 0.78 and 0.72 found in adult sheep.The results are consistent with a requirement for C20 and C22 polyunsaturated acids of the ω3 and ω6 series and some penetration of maternal fatty acids through the placenta. The resemblance between the fatty acid make‐up and composition of foetal and maternal phospholipids suggests the possibiligy of transference of intact or lyso‐phospholipids from the mother to the foetus through the placenta. However, such a possibility is counter‐indicated by consideration of previous work using labelled intermediates and by the mechanism of conversion of linoleic and linolenic acids requiring their CoA derivatives in the formation of the corresponding polyunsaturated C20 + C22 acids. Nevertheless, the sharp cut‐off of exogenous maternal fatty acids from the foetal triglycerides and their inclusion in the foetal phospholipids are not readily explainable.
Characteristics of pectinesterase isolated from bramley apple waste
Authors: King Karen 
Published: March 2007

AbstractPectinesterase was isolated from Bramley apple waste (peel, cores and offcuts) and its characteristics were determined. Optimum pectinesterase activity was obtained between pH 7.0 and 9.5 and activity was still detectable at 5.0. The enzyme was stable over a wide pH range in the presence of substrate, with over 90% of the activity retained after 5 min at pH 2.5. Pectinesterase activity increased with temperature up to 60°. Activation energy was calculated at 32 kJ mol−1 K−1 between 20 and 50°. Optimum activity was obtained in sodium chloride concentrations between 0.025 and 0.20 M with activity declining at higher salt concentrations. A Km of 0.044% apple pectin and a Vmax of 9.52 units g−1 DM were obtained for one specific enzyme extract.
Interrelationship of nitrogen nutrition with maize (Zea mays) grain yield, nitrogen use efficiency and grain quality
Authors: Tsai C Y , Dweikat I , Huber D M , Warren H L 
Published: March 2007

AbstractEight maize (Zea mays L) hybrids were grown under five N levels with or without the nitrification inhibitor, nitrapyrin (2‐chloro‐[6‐trichloro‐methyl] pyridine), to evaluate N interactions relative to yield performance, N use efficiency, grain protein concentration, and kernel texture. Results indicate that maize hybrids can be grouped into three categories based on grain yield: (1) low N‐responsive types which reach their maximum yield with 134 kg ha−1 of applied N; (2) intermediate types that respond to moderate N levels (134 to 201 kg N ha−1); and (3) high N‐responsive types that respond to higher levels of N (201 kg N ha−1). High N‐responsive types, in general, increased yield with nitrapyrin treatment at all levels of N. Crop N utilisation efficiency for high N‐responsive hybrids decreased, but was static for low N‐responsive hybrids as N fertiliser increased. In general, as the grain yield of a hybrid increased in response to N, the concentration of protein in the kernel increased; although grain yields and protein concentration are negatively correlated among hybrids. Increased kernel translucence, an indicator of kernel hardness induced by N fertiliser, correlated highly positive with zein proteins. Isoelectric focusing analysis showed that increases in zein were primarily due to a quantitative increase in α‐ and γ‐zein polypeptides. This study indicates that hybrids are different in their N requirements for maximum yield. Low N conditions not only restrict grain yield but also affect kernel textural quality.