EVALUATION OF PASSIVE TRANSFER IN NINE SPECIES OF CERVIDAE.

The reliability of packed cell volumes (PCV), total solids (TS), blood glucose (BG), γ-glutamyl transferase (GGT), and glutaraldehyde test in determining passive transfer of colostral immunoglobulins was investigated in nine species of cervids: axis deer (Axis axis), hog deer (Axis porcinus), sika deer (Cervus nippon), tufted deer (Elaphodus cephalophus), Père David's deer (Elaphurus davidianus), pudu (Pudu puda), sambar deer (Rusa unicolor), barasinga deer (Rucervus duvaucelli), and Eld's deer (Rucervus eldii). Individually the parameters evaluated were significant though imperfect predictors of passive transfer status in cervids. Interpreted collectively as a panel along with neonate condition, these tests were clinically helpful in diagnosing failure of passive transfer (FPT). Collectively interpreting test results as a panel along with clinical assessment of the animal is recommended. Some species-specific variations in TS, GGT, and glutaraldehyde test results were identified.

Glutaraldehyde test for the rapid diagnosis of pulmonary and extra-pulmonary tuberculosis in an area with high tuberculosis incidence.

BACKGROUND: Tuberculosis (TB) remains one of the leading causes of morbidity and mortality worldwide. The primary method for controlling TB is the rapid and accurate identification of infected individuals. Immune response exploitation represents one of the main methods used for early TB diagnosis; however, few studies have reported that whole blood originating from TB-infected patients gels faster in the presence of aldehyde than blood originating from healthy subjects, which is the focus of the current study. OBJECTIVES: The study objectives are to determine the diagnostic value of a glutaraldehyde test (GT) in pulmonary tuberculosis (PTB) and extra-pulmonary tuberculosis (EPTB) and to assess its performance compared with light-emitting diode fluorescence microscopy (LED-FM). MATERIALS AND METHODS: This study included 272 specimens (176 suspected PTB specimens and 96 suspected EPTB specimens). Of the 272 patients, 98 patients had TB infection confirmed by culture (64 PTB cases and 34 EPTB cases), and 174 patients had no TB infection. The gold standard technique (culture) was used as reference to verify the GT's performance. RESULTS: The GT showed a high sensitivity (96.9%) and specificity (82.1%) for PTB with a good positive predictive value (PPV = 75.6%) and negative predictive value (NPV = 97.9%). For EPTB, the GT showed a sensitivity of 91.2% and a specificity of 77.4%, with PPV = 68.9% and NPV = 94.1%. LED-FM had lower sensitivities for PTB (65.6%) and EPTB (42.1%) and an excellent specificity of 100%, with PPV = 100% and NPV = 100%. CONCLUSION: We concluded that GT is rapid, easy, simple and cost-effective and does not require qualified personnel with a specific background or sophisticated equipment like molecular biology or mycobacterium-specific genotyping techniques. These qualities make the GT attractive for use in low- and high-income countries in addition to other conventional methods, particularly culture, which continues to be the gold standard.

Glutaraldehyde test for the rapid diagnosis of pulmonary and extra-pulmonary tuberculosis in an area with high tuberculosis incidence

BACKGROUND Tuberculosis (TB) remains one of the leading causes of morbidity and mortality worldwide. The primary method for controlling TB is the rapid and accurate identification of infected individuals. Immune response exploitation represents one of the main methods used for early TB diagnosis; however, few studies have reported that whole blood originating from TB-infected patients gels faster in the presence of aldehyde than blood originating from healthy subjects, which is the focus of the current study. OBJECTIVES The study objectives are to determine the diagnostic value of a glutaraldehyde test (GT) in pulmonary tuberculosis (PTB) and extra-pulmonary tuberculosis (EPTB) and to assess its performance compared with light-emitting diode fluorescence microscopy (LED-FM). MATERIALS AND METHODS This study included 272 specimens (176 suspected PTB specimens and 96 suspected EPTB specimens). Of the 272 patients, 98 patients had TB infection confirmed by culture (64 PTB cases and 34 EPTB cases), and 174 patients had no TB infection. The gold standard technique (culture) was used as reference to verify the GT's performance. RESULTS The GT showed a high sensitivity (96.9%) and specificity (82.1%) for PTB with a good positive predictive value (PPV = 75.6%) and negative predictive value (NPV = 97.9%). For EPTB, the GT showed a sensitivity of 91.2% and a specificity of 77.4%, with PPV = 68.9% and NPV = 94.1%. LED-FM had lower sensitivities for PTB (65.6%) and EPTB (42.1%) and an excellent specificity of 100%, with PPV = 100% and NPV = 100%. CONCLUSION We concluded that GT is rapid, easy, simple and cost-effective and does not require qualified personnel with a specific background or sophisticated equipment like molecular biology or mycobacterium-specific genotyping techniques. These qualities make the GT attractive for use in low- and high-income countries in addition to other conventional methods, particularly culture, which continues to be the gold standard.

Hemoglobin-based oxygen carrier microparticles: synthesis, properties, and in vitro and in vivo investigations.

Bovine hemoglobin microparticles (Hb-MPs) as suitable oxygen carriers are fabricated easily by three key steps: coprecipitation of Hb and CaCO(3) to make Hb-CaCO(3)-microparticles (Hb-CaCO(3)-MPs), cross-linking by glutaraldehyde (GA) to polymerize the Hb and dissolution of CaCO(3) template to obtain pure Hb-MPs. The Hb entrapment efficiency ranged from 8 to 50% corresponding to a hemoglobin quantity per Hb-MP of at least one-third of that in one erythrocyte. The Hb-MPs are spherical, with an average diameter of 3.2 µm and high oxygen affinity. The methemoglobin level was increased after preparation, but can be reduced to less than 7% with ascorbic acid. Phagocytosis assays showed low immunogenicity of Hb-MPs if the particles were cross-linked with low concentration of GA and treated with sodium borohydride. Magnetite-loaded Hb-MPs circulated up to 4 days after intravenous application.

The interaction of DNR and glutaraldehyde with cell membrane proteins leads to morphological changes in erythrocytes.

In this study, the effects of DNR and glutaraldehyde on isolated erythrocyte membrane proteins were examined. For this purpose, SDS-gel electrophoresis was carried out. Additionally, analyses of the disturbances in erythrocyte shape and size, accompanied by the application of flow cytometry and microscopy examination, were undertaken. The amount of DNR linked to erythrocyte cell membranes was measured by a fluorimetric technique. It was observed that glutaraldehyde caused in concentration dependent manner an increase of percent of DNR linked to cell membrane proteins. After this incorporation, perturbations in the protein content of cell membranes were observed. The protein aggregates and changes in the level of spectrin, actin and band 3 protein were noted. Due to the changes in spectrin, which is mainly responsible for maintenance of the discocyte shape of erythrocytes, flow cytometry and microscopy techniques were used to control the size and shape of erythrocytes after treatment with DNR and glutaraldehyde. The disturbances in the shape and size of erythrocytes were observed for all tested concentrations of glutaraldehyde. For all tested concentrations of glutaraldehyde, the changes were statistically significant.

Synthesis and physicochemical characterization of a series of hemoglobin-based oxygen carriers: objective comparison between cellular and acellular types.

A series of hemoglobin (Hb)-based O(2) carriers, acellular and cellular types, were synthesized and their physicochemical characteristics were compared. The acellular type includes intramolecularly cross-linked Hb (XLHb), polyoxyethylene (POE)-conjugated pyridoxalated Hb (POE-PLP-Hb), hydroxyethylstarch-conjugated Hb (HES-XLHb), and glutaraldehyde-polymerized XLHb (Poly-XLHb). The cellular type is Hb-vesicles (HbV) of which the surface is modified with POE (POE-HbV). Their particle diameters are 7 +/- 2, 22 +/- 2, 47 +/- 17, 68 +/- 24, and 224 +/- 76 nm, respectively, thus all the materials penetrate across membrane filters with 0.4 microm pore size, though only the POE-HbV cannot penetrate across the filter with 0.2 microm pore size. These characteristics of permeability are important to consider an optimal particle size in microcirculation in vivo. POE-PLP-Hb ([Hb] = 5 g/dL) showed viscosity of 6.1 cP at 332 s(-1) and colloid osmotic pressure (COP) of 70.2 Torr, which are beyond the physiological conditions (human blood, viscosity = 3-4 cP, COP = ca. 25 Torr). XLHb and Poly-XLHb showed viscosities of 1.0 and 1.5 cp, respectively, which are significantly lower than that of blood. COP of POE-HbV is regulated to 20 Torr in 5% human serum albumin (HSA). HES-XLHb and POE-HbV/HSA showed comparable viscosity with human blood. Microscopic observation of human red blood cells (RBC) after mixing blood with POE-PLP-Hb or HES-XLHb disclosed aggregates of RBC, a kind of sludge, indicating a strong interaction with RBC, which is anticipated to modify peripheral blood flow in vivo. On the other hand, XLHb and POE-HbV showed no rouleaux or aggregates of RBC. The acellular Hbs (P(50) = 14-32 Torr) have their specific O(2) affinities determined by their structures, while that of the cellular POE-HbV is regulated by coencapsulating an appropriate amount of an allosteric effector (e.g., P(50) = 18, 32 Torr). These differences in physicochemical characteristics between the acellular and cellular types indicate the advantages of the cellular type from the physiological points of view.

Evaluation of a whole blood glutaraldehyde coagulation test for the detection of failure of passive transfer in calves.

The accuracy of a commercially available whole blood glutaraldehyde clot test in the detection of failure of passive transfer (serum immunoglobulin [Ig]G1 < 1,000 mg/dL) in neonatal calves was evaluated. Serum samples were obtained from 242 calves ranging in age from 1 to 8 days, and comparisons were made with serum IgG1 concentrations determined by radial immunodiffusion. Both the sensitivity and specificity of the currently marketed whole blood glutaraldehyde clot test are inadequate for routine diagnostic use. Concerns regarding test sensitivity are the most problematic. Sensitivity varied from 0.41 to 0.00, depending on the choice of test endpoint. Specificity varied from 0.85 to 1.00, depending on the choice of test endpoint. Regression analysis demonstrated that the relationship between serum IgG1 concentration and the glutaraldehyde clot results, although significant (P < .10), was of negligible biological relevance (r2 = .034).

Radioimmunoassay for ceronapril, a new angiotensin-converting enzyme inhibitor, and its application to a pharmacokinetic study in healthy male volunteers.

Ceronapril is a member of a new chemical class of angiotensin-converting enzyme inhibitors being developed by The Bristol-Myers Squibb Pharmaceutical Research Institute. A radioimmunoassay (RIA) has been developed for the measurement of ceronapril in biological fluids. The RIA has a range of 0 to 500 ng/ml and has the sensitivity to detect 1.0 ng/ml of ceronapril. Satisfactory zero binding and sensitivity were obtained after a 2-h incubation at room temperature or overnight at 4 degrees C. Separation of the antibody-bound and free radiolabel was achieved by employing polyethylene glycol-goat anti-rabbit gamma-globulin separant. A quantitative recovery of the exogenous analyte was obtained at all concentrations of ceronapril tested. Intraassay coefficients of variance (CV's) were 3.9% and 4.6% for the low and medium controls, respectively. A highly significant statistical correlation between RIA and [14C]TLRC was observed for both plasma and urine samples. Clinical samples from the ascending dosage studies have been analyzed by the ceronapril RIA. The maximum concentration and the area under the plasma concentration-time curve did not increase in a dose-proportional manner for doses above 100 mg.

Distribution, metabolism, and excretion of [14C] glutaraldehyde.

Radiolabeled glutaraldehyde (GA) was infused into rats in order to determine its distribution between cellular and humoral fractions of the blood, its potential metabolism by RBC's, its rate of excretion, and the nature of its urinary products. This study demonstrated that [14C]GA was distributed between the RBC's and plasma at a ratio greater than 1. Although the absolute counts of both fractions dropped 80% over 3 days, the percentage bound or incorporated increased over time. Despite the extensive uptake by RBC's these cells were unable to metabolize GA to CO2. Urinary excretion of the radiolabel was rapid; the predominant form in the urine was less than 1 kDa in size. All evidence suggested that it was not native GA. We conclude that the RBC's can incorporate GA, but can not metabolize it completely to CO2. Nevertheless, much of the infused GA was rapidly converted to nonreactive metabolites and eliminated by the kidney.