Effects of purified bovine whey factors on cellular immune functions in ruminants

Retinoid X receptors (RXRs) form heterodimers with thyroid hormone receptors (TRs). RXRs increase DNA binding affinity of TRs and T3-mediated transactivation on positive T3 response elements (TREs). However, the role of RXRs on negative TREs, and the relation of RXRs to the dominant negative effect of mutant TRs, are not defined. To clarify the function of RXRs on negative TREs, we performed transient cotransfection studies using the rat glycoprotein hormone alpha promoter fused to luciferase gene (alphaLuc), and human TRH promoter fused to luciferase gene (TRH-Luc) as reporters. We found that the JEG-3 cell-alphaLuc system was very sensitive to TR regulation. Using TRbeta1 wild-type (WT) expression vector, 6.2 ng/well (170 ng/10 cm dish), and 0.2 ng/well (11 ng/10 cm dish) caused maximal, and half maximal, inhibition of Luc activities in the presence of 1 nM T3. A T3 dose dependent inhibition study was also performed. From these studies, we determined that the appropriate conditions in which to study alphaLuc transactivation, in a linear portion of the dose response curve, was using 0.8 ng/well TRbeta1 expression vector and 0.1 nM T3. Under these conditions, TRbeta1 mutant R316H (GH), but not G345R (Mf), showed a weak dominant negative effect at a 1:1 ratio in the presence of 0.1 nM T3 although neither mutant had detectable T3 binding affinity. Moreover this dominant negative effect of R316H on the alphaLuc reporter was enhanced in the presence of RXRgamma. Mutant G345R showed a stronger dominant negative effect than did R316H when using a double palindromic TRE fused to herpes simplex thymidine kinase-Luc reporter as a positive TRE. These results conform to the clinical features of R316H which is associated with apparent pituitary resistance of thyroid hormone (PRTH). Mutant R316H also showed a weak dominant negative effect with TRH-Luc at a 1:1 ratio in the absence or presence of RXRgamma. However RXRgamma did not enhance the dominant negative effect as it did using alphaLuc reporter gene. Electrophoretic gel mobility shift assay (EMSA) showed that RXR alpha augmented the DNA binding affinity of wild type and R316H TRs as heterodimers on the previously reported negative TREs of glycoprotein hormone alpha promoter, suggesting that RXR does not produce its response by removing TRs from these TREs. RXR alpha augmented DNA binding affinity of TRbeta1WT, and R316H showed a weaker heterodimer band than did the wild type in EMSA. Using the TRH-Luc reporter, basal activity was increased by wild type TRbeta1. However a TRbeta1 DNA binding domain mutant, (C127S) which can not bind to DNA, did not increase the basal activity. This indicates that DNA binding of the TR is required for increasing basal activity of TRH promoter. These results indicate that (1) RXR-TR heterodimers play a role in basal transactivation and T3 suppression of negatively regulated genes, and (2) RXRs increase the dominant negative effect of some mutant TRs on specific negative TREs. (3) This effect occurs without removing TRs from the TRE. (4) The differential dominant negative effect of mutant R316H (negative TRE > positive TRE) may explain, at least in part, the presentation of R316H as PRTH. (5) Augmentation of basal activity by wild type TRs on a negative TRE requires DNA binding.     

An orally administered growth factor extract derived from bovine whey suppresses breath ethane in colitic rats

The aim of the present study was to assess the in vivo error of the method as well as the effect of thresholding when obtaining and evaluating standardized periapical radiographs for computer-assisted densitometric image analysis (CADIA). Twenty healthy volunteers participated in an experimental gingivitis study in which neither mechanical nor chemical plaque control was performed for 21 days. Two pairs of standardized periapical radiographs were taken at days 0 (baseline) and 21 (follow-up), one from a maxillary area (15 volunteers) and one from a mandibular molar/premolar area (17 volunteers). Each baseline radiograph was digitized and its image displayed on a monitor. The follow-up radiograph was then superimposed and digitized as well. After gray level correction, subtraction radiographic images were produced. The difference in gray level between the baseline and the follow-up image was calculated within each region of interest (ROI) at each picture point (pixel). In bone ROI, changes in density reflected the amount of change due to methodological errors plus the basic bone remodeling over 3 weeks. For gingival ROI, changes in density reflected the methodological error plus a possible change in soft tissue density during the experimental gingivitis. Within all of the ROI, some pixels indicated a change in gray level. A change in gray level was then thresholded; i.e., only changes >5 and then >10 gray levels were registered and used for calculation of the CADIA values. With a threshold of 5, 44/45 maxillary bone ROI and 60/66 mandibular bone ROI showed a change in density, while 41/45 maxillary gingiva ROI and 26/66 mandibular gingiva ROI indicated a change in density. With a threshold of 10, 16/45 maxillary bone ROI and 12/66 mandibular bone ROI indicated a change in density, while 13/45 maxillary gingiva ROI and 1/66 mandibular gingiva ROI indicated a change. The amounts of changes in density calculated in the various ROI were low even when applying no threshold, ranging from -0.279 to 0.621. Applying a threshold of 5, the CADIA values ranged from -0.234 to 0.727. With a threshold of 10, the changes in density ranged from -0.318 to 0.133. In vivo, CADIA of standardized radiographs indicated change in density due to methodological errors. Application of thresholds may avoid false-positive diagnoses. When applying CADIA in clinical research, the range of change to be expected due to methodological limitations as well as the threshold for true change should be evaluated. These thresholds may differ in various areas of the mouth, i.e., bone or gingival, maxillary/mandibular, anterior/posterior ROI.     

Biocompatibility of cellulose sponge with bone

The purpose of this study was to investigate the biocompatibility of viscose cellulose sponge (VCS) with bone. Twenty-five Sprague-Dawley rats were used for the study. After curettage of the bone marrow from both femoral cavities, VCS (15 x 1 x 1 mm) was implanted into one femur, leaving the contralateral side empty as a control. The rats were killed 1-6 weeks after curettage, and bone formation inside the sponge was assessed by light-microscopic examination and histomorphometric assessment. Whereas normal bone formation in rat femoral cavity took place in 2 weeks after curettage, 4 weeks were needed for bone formation in the cellulose sponge. VCS is a compatible matrix for osseous tissue ingrowth and it may be useful as a scaffold for bone tissue engineering in experiments and possibly also in clinical practice.     

Combined inhibitory effect of bovine immune whey and peroxidase-generated hypothiocyanite against glucose uptake by Streptococcus mutans

A case of chronic ulcer of the lower lip, which persisted for three years and resisted all locals treatments, is presented. Recovery was achieved with excision. The specimen was examined macroscopically, and showed a prominent inferior labial artery or calibre-persistent artery, an uncommon entity. Patient-literature was reviewed.     

Contribution to phosphorus behaviour in steelmaking with sponge iron

Experiments were conducted in a 70 t UHP electric arc furnace to study the phosphorus behaviour in steelmaking with sponge iron. The proportion of sponge iron in the metallic feed was varied from 52 to 95.4 wt.%. The results are used to show the effect of the sponge iron proportion in the charge on the phosphorus content of the bath. They are also utilized to illustrate the effects of the composition, temperature and theoretical optical basicity of the slag on the phosphate capacity and to investigate the kinetics of dephosphorization. It is found that both initial and final phosphorus contents of the bath decrease almost linearly with increasing sponge iron proportion in the charge. Using regression analysis, linear equations were obtained which show the change of the phosphate capacity of a slag, at a given temperature, with the total lime content as well as with the theoretical optical basicity. Based on these equations, a functional relationship was derived which shows the effect of simultaneous variation of total lime content and temperature of a slag on its phosphate capacity. A similar equation is given for the influence of simultaneous change of theoretical optical basicity and temperature on the phosphate capacity of a slag. The effect of the compositions of the slag and metal on the phosphorus distribution ratio is expressed by an equation. The kinetics of dephosphorization are investigated, assuming that the mass transfer of phosphorus in the metal is the rate controlling step. In the present investigation the activation energy of dephosphorization is equal to 38 kJ/mol and the mass transfer coefficient of phosphorus at 1600°C is 0.0135 cm/s.     

Porous-type tracheal prosthesis sealed with collagen sponge

BACKGROUND: Reconstruction of a long section of the trachea is clinically problematic. Tracheal reconstructions using prostheses have met with limited success due to local infection, hemorrhage, luminal stenosis and prosthesis dislocation. METHODS: We have designed a porous type of tracheal prosthesis in which the mesh is sealed with collagen sponge. We used this prosthesis (50 mm in length) to reconstruct the cervical trachea in 10 mongrel dogs and evaluated its efficacy. RESULTS: One dog died due to an accident with anesthesia at 6 weeks and 1 of suffocation at 10 weeks. The other 8 dogs had an uneventful postoperative course until they were killed between 6 and 24 months after implantation. At sacrifice, all the prostheses had become completely incorporated into the host. Microscopic examination revealed advanced formation of a new epithelial lining in 1 dog at 6 months, and a confluent epithelial lining was observed in another dog at 12 months. Central stenosis was not significant in any of the animals. CONCLUSIONS: This tracheal prosthesis gives good results in canine tracheal reconstruction, and appears very promising for the clinical repair of tracheal defects.     

Primary culture and expression of cytokine mRNAs by lipopolysaccharide in bovine Kupffer cells

To clarify the mechanisms of the antiepileptic activity of phenytoin (PHI), the effects of PHT on extracellular and total levels of monoamines (dopamine and serotonin), in rat striatum and hippocampus were studied. The plasma concentrations of PHT associated with therapeutic activity did not affect striatal and hippocampal extracellular levels of monoamines, whereas supratherapeutic concentrations of PHT decreased striatal and hippocampal extracellular levels of monoamines, in a concentration dependent manner. Toxic concentrations of PHT produced generalized seizures 'paradoxical intoxication' and an initial drastic decrease in striatal and hippocampal extracellular levels of monoamines before seizure onset, whereas the extracellular monoamines levels increased after seizures. In addition, the therapeutic concentrations of PHT did not affect monoamine turnover, whereas supratherapeutic concentrations of PHT inhibited monoamine turnover. These results suggest that monoaminergic transmission may not be involved in the antiepileptic mechanism of action of PHT, and that dysfunction of monoaminergic transmission can produce generalized tonic-clonic convulsions. Thus, the present study suggests that 'Paradoxical Intoxication' induced by toxic concentrations of PHT, at least partially, can be mediated by hypo-monoaminergic function in the brain.     

Procollagen and collagen produced by normal bovine corneal stroma fibroblasts in cell culture

Procollagen and collagen were isolated from the culture medium of normal bovine corneal stromal fibroblasts. DEAE-cellulose chromatography was used to separate the collagen molecules from the different procollagens present. One collagen and four procollagen peaks were isolated and biochemically characterized. All the procollagen fractions and the collagen fraction yielded, after limited pepsin or chymotrypsin digestion followed by CNBr digestion, molecules that correspond to (alpha 1)2 alpha 2 exclusively. Thus only type I collagen is found in the growth medium of of bovine corneal stromal fibroblast cultures. Each of the individual procollagen peaks contained pro-alpha chains having molecular weights of 120,000, 150,000, 165,000, 180,000, and 190,000 daltons, according to their elution position on DEAE-cellulose. The presence of type I procollagen molecules having pro-alpha chains of 165,000, 180,000, and 190,000 daltons has not previously been reported and probably represents higher-molecular-weight precursor intermediates. The amino acid compositions of the different procollagen fractions are unique, and each contains relatively large amounts of cysteine and tryptophan. Carbohydrate analysis, cyanogen bromide peptide analysis, electron microscopy of SLS-crystallities, and SDS-polyacrylamide gel electrophoresis were used to further characterize the procollagen and collagen molecules.     

Development of in vitro derived bovine embryos following pronuclear transplantation and in vitro culture

This study was designed to evaluate the survival and development of in vitro derived bovine embryos following pronuclear transplantation and in vitro embryo culture. Bovine zygotes were produced by in vitro maturation and in vitro fertilization. Pronuclei were removed by micromanipulation and either transferred back to the same cell (Group 1) or into a previously enucleated zygote (Group 2) by electrofusion. Micromanipulated and non-micromanipulated (Group 3, control) zygotes were co-cultured with oviductal cells in a sealed modular chamber filled with 5% CO2, 5% O2 and 90% N2 at 39 degrees C for 7-8 days. Fusion rates were similar for Groups 1 and 2 (90.7 and 85.1%, respectively, P > 0.05). The percentage of embryos that cleaved was not different for Groups 1 (82.0%), 2 (90.0%) and 3 (76.9%, P > 0.05). Also, the percentage of embryos developing to the compact morula or blastocyst stage was similar (25.6, 22.5 and 22.3%, respectively, for Groups 1, 2 and 3, P > 0.05). The results of this experiment are the first to demonstrate that pronuclear transfer can be carried out successfully using bovine embryos derived from in vitro oocyte maturation and in vitro fertilization. In addition, pronuclei can be transferred from one bovine embryo to another and the reconstructed embryos develop to the compact morula and blastocyst stage in vitro. This technique, used in combination with oocyte retrieval by ultrasound-guided follicular aspiration and embryo transfer, offers the potential to study cytoplasmic inheritance in cattle directly, and to evaluate the effect of cytoplasmic inheritance on traits of economic importance.     

The whey proteins of the milk of red deer (Cervus elaphus L.). A homologue of bovine beta-lactoglobulin

Increasing plasma free fatty acids decreased the degree of glycogen depletion, and increased the citrate concentration, in slow-red (soleus) and fast-red (deep portion of vastus lateralis) muscle during exercise (approx. 50% depletion of glycogen, as against 75% in control animals). There was no effect in fast-white muscle (superficial portion of vastus lateralis). Glycogen concentration in the liver decreased by 83% in controls, but only by 23% in animals with increased free fatty acids during exercise. The decreased glycogen depletion may be partly explained by the findings that (a) plasma-insulin concentration was two- to three-fold higher in animals with increased plasma free fatty acids and (b) the exercise-induced increase in plasma glucagon was lessened by increased free fatty acids. Blood glucose was higher in the animals with increased free fatty acids after the exercise. The rats with increased plasma free fatty acids utilized approx. 50% as much carbohydrate as did the controls during the exercise.     

Thermodynamic stability of bovine alpha-crystallin in its interactions with other bovine crystallins

Light scattering measurements were performed on dilute solutions of alpha-crystallin mixed with different combinations of beta H, beta L and gamma-fractions of bovine lens crystallins. Light scattering intensities were obtained as a function of scattering angle, concentration and temperature. The temperature dependence of the second virial coefficients was used to obtain partial molar enthalpy and end entropy of solutions. The difference between the thermodynamic parameters of the crystallin mixtures and those of the weighted averages of the individual components yielded the excess enthalpy and entropy functions of the solutions. Both the excess enthalpy and entropy functions indicated that thermodynamic stability of alpha-crystallin is progressively enhanced by its interactions with gamma [symbol: see text] (beta H + gamma) [symbol: see text] (beta H + beta L + gamma) crystallins. The last two combinations showed negative values both for excess enthalpy as well for excess entropy of solutions. Other combinations demonstrated increasing positive values. This implies that the combination of all four crystallins in the vertebrate lens enables the best solvation property as well as the best packing as opposed to any other single or combinatorial arrangements of crystallins. Similar conclusions have been obtained in the past from water and other vapor sorption studies.     

超声溶解电位滴定法测定海绵钛中氯

使用超声震荡辅助氢氟酸、硝酸溶解海绵钛试样,加入硼酸生成氟硼酸掩蔽氢氟酸,用硝酸银溶液进行电位滴定,测定了海绵钛中微量氯。由于采用快速的等间隔滴定和在滴定完成后40 s记录电位值,因此减少残余电位漂移对测定的影响。滴定终点采用电位二阶导数为零的精确方法判断,测定误差小,结果更可靠。试验表明,基体钛对氯的测定设有干扰,试样中S2-,Br-,I-,CN-对测定有干扰,但存在量很低,影响可以忽略。用本法测定2个海绵钛样中氯,结果与国家标准方法(硫化银分光光度法)相符,相对标准偏差分别为1.48%和2.59%(n     

On sulphur behaviour in steelmaking with sponge iron in an electric arc furnace

The sulphur behaviour in steelmaking by melting of charges containing from 38.7 to 95.4 wt.% sponge iron in the metallic input in a 70 t UHP electric arc furnace is investigated. The effect of both slag composition and temperature on the sulphide capacity of the slag and on the sulphur distribution ratio between slag and metal is studied. The kinetics of desulphurization are also treated. Regression analysis shows that the logarithm of sulphide capacity varies linearly with total lime equivalent, theoretical optical basicity and logarithm of slag basicity. Equations relating the sulphide capacity to these parameters as well as to the temperature are given. Based on these equations, functional relationships for the calculation of sulphur distribution between slag and metal are derived. Metallurgical reactions for metal desulphurization by slag and equilibrium relationships are given from which the slag-metal sulphur partition ratio can be calculated. In all cases, the results of the calculations are in line with the experimental data. The rate of desulphurization is expressed by an equation based on the boundary layer diffusion theory. Under the conditions of the present investigation, the activation energy of desulphurization is found to be 39 kJ/mol and the mass transfer coefficient of sulphur at 1600°C is equal to 0.014 cm/s.     

Some metallurgical aspects of steelmaking with sponge iron in electric arc furnaces

Test charges containing sponge iron proportions varying from 38.7–95.4 wt% of the metallic input were melted in a UHP electric arc furnace with a design capacity of 70 tons per heat. After melting started, samples of molten metal and the corresponding slag were taken simultaneously at different time intervals and analysed. The time dependence of the chemical composition of the metal and slag as well as the variation of the temperature of the melt with time are given. Thermodynamic calculations using different methods for finding the activity of ferrous oxide show that there is a linear relationship between a_(FeO) and the total ferrous oxide content in the slag. It is also found that the total ferrous oxide content in the slag and the oxygen concentration in the metal vary linearly with the iron proportion from sponge iron in the metallic input. The activity of ferrous oxide decreases with increasing slag basicity due to the formation of calcium ferrites. The relationship between the oxygen concentration and the reciprocal of the carbon content in liquid steel is almost linear. A formula showing the influence of some important factors on the oxygen content in molten steel is given. The effect of sponge iron on the sulphur concentration in the steel is also investigated. The present results indicate that the sulphur content in the steel can be reduced from 0.02 to 0.004 wt% by increasing the iron proportion from sponge iron in the metallic input from 35 to 95 wt %. Using the boundary layer diffusion model, it is found that the rate of decarburization of a steel bath is enhanced by increasing the sponge iron proportion in the metallic input. The activation energy of the decarburization reaction is found to be 56.9 kJ/mol and the mass transfer coefficient of carbon has a value of 0.0161 cm/s at 1 600°C. In a way similar to that used for decarburization, it is also found that the rate of oxidation of manganese dissolved in the bath is enhanced by increasing the sponge iron proportion in the metallic input.     

Medullary sponge kidney (MSK) in association with glomerulonephritis (GNP)]

BACKGROUND: Medullary sponge kidney (MSK) is a congenital, non-hereditary anomaly characterized by dilation of the precalix ducts. It is often associated with other diseases, and its symptoms are linked to frequent complications, such as sepsis of the urinary system and the renal colic of nephrolithiasis. However, MSK is rare in association with glomerulonephritis (GNP), as can be seen from medical literature data that shows only one case (reported in 1974) in which MSK with a concomitant focal sclerosing glomerulonephritis was noted. In this report, we describe a case of MSK that was found after a routine renal echograph and biopsy of a mesangial GNP previously diagnosed at another center. The echographic results showing typical MSK was confirmed by means of RX urography and renal TC. The MSK-glomerulonephritis combination appears to be completely random, because the involved structures (the collecting duct and the glomerule) have different embryologic origin. Therefore, a pathogenic hypothesis common to the two anomalies seems rather unlikely. CONCLUSIONS: Although MSK is a usually benign disease, because a compromise of glomerular filtration that occurs in approximately 10% of cases, proper diagnosis is important to allow prophylactic intervention aimed at the reduction of complications that could reduce overall renal function when associated with other nephropathies. Renal echography is an important diagnostic tool that may reveal more cases of MSK than previously reported.     

The role of commercial bovine serum albumin preparations in the culture of one-cell rabbit embryos to blastocysts

Normal bovine serum albumin (BSA) in a complete medium without energy substrates promoted growth of 1-cell embryos to hatched blastocysts. Defatted charcoal-treated BSA did not promote growth to the blastocyst stage but the addition of pyruvate or palmitic and oleic acids allowed blastocyst growth but not blastocyst hatching. Sodium dodecyl sulphate-gel electrophoresis showed that both the normal and defatted BSA samples were heavily contaminated by proteins other than albumin. Fractionation of the normal BSA on Sephadex G-200 indicated that the property of promoting complete blastocyst hatching was not due to the albumin but was associated with the higher mol. wt fraction of the BSA. Extraction of normal BSA with chloroform appeared to destroy the hatching-promoting ability as neither the residue after extraction nor defatted BSA to which the organic extractate had been added promoted complete blastocyst hatching. It is concluded that commercial BSA may have at least two effects on blastocyst growth: (1) energy provision via albumin-bound fatty acids, and (2) promotion of blastocyst hatching by a non-albumin component.     

Purification and characterization of novel whey glycoprotein WGP-88 which binds to a monoclonal antibody to PAS-4 glycoprotein in the bovine milk fat globule membrane

A monoclonal antibody to the PAS-4 glycoprotein (78 kDa) of the bovine milk fat globule membrane (MFGM) specifically recognized PAS-4, and was named KAS4. A component recognized by KAS4 was found in whey protein, this being a glycoprotein of 88 kDa by SDS-PAGE and named WGP-88. WGP-88 was purified and characterized in comparison with PAS-4. WGP-88 had apparent pI values of 6.45 and 6.39, while those of PAS-4 were 7.39 and 7.35. Neuraminidase digestion shifted the pI values of WGP-88 to 6.57 and of PAS-4 to 7.52. WGP-88 was rich in polar amino residues (44.9 mol%), while PAS-4 was abundant in nonpolar amino acid residues (48.7 mol%). WGP-88 contained 17.1% of carbohydrate and PAS-4 had 7.2%. The results of reductive hydrolysis, N-glycanase digestion, and a lectin blot analysis suggested that N- and O-linked sugar chains were contained in both glycoproteins. WGP-88 and PAS-4 had a different N-terminal amino acid sequence. WGP-88 and PAS-4 respectively inhibited competitively the binding of KAS4 to PAS-4 and WGP-88. Our studies revealed WGP-88 recognized by KAS4 mAb to be a novel whey protein and to have different biochemical properties from those of PAS-4.