Effects of the composition of the basal diet on the evaluation of mineral phosphorus sources and interactions with phytate hydrolysis in broilers.

The objectives of this study were to determine the availability of P from mineral phosphate sources by using different basal diets and measurement of P retention and prececal (pc) P digestibility as well as pc myo-inositol phosphate (InsP) degradation in broilers. Semi-synthetic and corn-soybean meal-based basal diets were used in experiment 1, and corn-based and wheat-based basal diets were used in experiment 2. Anhydrous monosodium phosphate (MSPa) or monocalcium phosphate monohydrate (MCPh) was supplemented to increment the P concentration by 0.05, 0.10, and 0.15% or by 0.075 and 0.150% in experiments 1 and 2, respectively. Titanium dioxide was used as an indigestible marker. Diets were pelleted through a 3-mm screen. In experiment 1, retention was measured based on total excreta collection from 20 to 24 d of age using 7 replicated birds per diet. In experiment 2, digesta from the terminal ileum was collected from 22-d-old broilers penned in groups of 19 with 5 replicated pens per diet. The P retention response to supplemented MSPa did not differ between the 2 basal diets in experiment 1. The response in pc P digestibility to MCPh supplements also did not differ between the 2 basal diets in experiment 2, as calculated by linear regression analysis. Hydrolysis of InsP6 measured on both the excreta and pc levels was high in the basal diets without a mineral P supplement. Mineral P supplementation significantly decreased (P < 0.05) InsP6 hydrolysis from the InsP-containing diets in both experiments. Thus, the choice of the basal diet did not affect the evaluation of the supplemented mineral P source. However, calculated values for mineral P sources need to be adjusted for the decline in hydrolysis of InsP contained in the basal diet that results from the P supplement.

Transgenic mice expressing a truncated Peromyscus leucopus TNF-alpha gene manifest an arthritis resembling ankylosing spondylitis.

Several studies have implicated tumor necrosis factor-alpha (TNF-alpha) in autoimmune diseases, such as rheumatoid arthritis (RA). To elucidate further the role of TNF-alpha in inflammatory arthritis, we generated transgenic mice harboring a truncated Peromyscus leucopus TNF-alpha (Pe-TNF) gene. An arthritic phenotype closely resembling human ankylosing spondylitis was observed only in transgenic lines expressing the Pe-TNF transgene at the mRNA level. We characterized the arthritic phenotype in detail by radiographic and histologic techniques. It consisted of severe axial skeletal kyphosis and ankylosis, accompanied by an inflammatory and fibrotic process at the end plates and enthesis. Peripheral joint lesions were absent in mice expressing the P. leucopus TNF-alpha gene, in contrast to the RA-like phenotype described in transgenic mice expressing a truncated human TNF-alpha gene. The Pe-TNF transgenic mouse model provides a unique opportunity to explore potential mechanisms whereby TNF-alpha may initiate an autoimmune arthritis resembling ankylosing spondylitis.

A warm-water perfusion plaque for hyperthermic treatment of intraocular tumors.

This paper describes the design and performance of a plaque aimed at delivering hot-water hyperthermia to intraocular tumors. The plaque transfers heat into the tissues via conduction. Maximum temperature cannot exceed inflow water temperature. The high water flow rate provided by the hyperthermia unit guarantees an excessive power output at a constant temperature level, resulting in a homogeneous temperature distribution in the tissue. Heat distribution over the plaque surface was shown to be uniform. System performance has been studied on enucleated human as well as on living rabbit eyes. Thermal mapping was done in two and one planes, respectively, and the spatial distribution was calculated. Results promise therapeutic temperature levels to a depth exceeding 10 mm.

Major histocompatibility complex class I genes of Peromyscus leucopus.

Class I genes of the Peromyscus leucopus major histocompatibility complex (MhcPele) were examined by Southern blot hybridization, genomic cloning, and DNA sequencing. At least three distinct subtypes of Pele class I genes were discerned, which we have designated Pele-A, B, and C. The nucleotide sequences of exon 5-containing regions (encoding the transmembrane domain) suggested that Pele-A genes are homologs of mouse H-2K, D, L, and Q genes and that Pele-B genes correspond to mouse Tla genes. The Pele-C genes appeared similar to mouse M1 genes. The number of unique genes in each subtype cloned from an individual P. leucopus were 20 for Pele-A, 13 for Pele-B, and 2 for Pele-C. Three genomic clones showed cross-hybridization to both Pele-A and Pele-B gene-specific probes. Six genomic clones remained unclassified as they did not cross-hybridize to exon 5-containing probes from Pele-A, B, or C genes. The homology between the transmembrane domains of Pele class I gene subtypes was found to be similar to that observed between the transmembrane domains of H-2 subtypes (or groups). Interspecific similarity of exon 5 was found to be 81%-88% between Pele class I genes and their H-2 counterparts.

A human-human hybridoma secreting anti-HLA class II antibody.

In this report, we describe the production and characterization of the first human-human hybridoma secreting antibody to HLA Class II determinants. The hybridoma (GMEC101), which has been stable in tissue culture for greater than 20 mo, secretes 10 to 50 micrograms/ml of IgM-kappa antibody. This antibody binds to a wide range of human cell lines, but not to the HLA-A,B,C, and DR-negative K562 cell line. Functionally, GMEC101 strongly inhibits a unidirectional mixed lymphocyte reaction (MLR) at the level of the stimulator cell. Neither the cellular ELISA binding nor the MLR inhibition is lost after a triple platelet absorption (which removes Class I but not Class II activity). Because the binding and MLR blocking show no correlation with the known DR or DQ specificities, we suggest that GMEC101 may be detecting a novel HLA Class II determinant.

Detection of antibodies to cell surface antigens by a simplified cellular ELISA (CELISA).

An improved method for screening human hybridoma antibodies to cell surface antigens is described. The following modifications have been developed: rapid expansion of desired screening cell types by EBV transformation; use of only 5 X 10(4) cells/well; elimination of the need for glutaraldehyde fixation; elimination of the requirement for PLL to attach cells to microplates; preparation of a large number of plates which can be stored at 4 degrees for 3 months; Protein A-peroxidase ELISA assay yielding excellent replicates, low background "noise", and high OD readings for positive wells. The techniques we have developed should greatly simplify and shorten the assay procedures for detecting human antibodies to a variety of cell surface antigens.

Results of a phase II study of the antidepressant effect of rolipram.

The antidepressant effect of rolipram, believed to be based on a new mechanism of action, was investigated in an open phase II study in 10 depressive patients, most of whom had been refractory to previous antidepressant therapy. Five patients displayed a good to very good improvement of their depressive condition. Four patients failed to show any substantial improvement, and therapy had to be withdrawn in one case due to deterioration of the condition. In most cases, the antidepressant effect of the trial preparation became noticeable after 2-4 days of treatment only. Basing on the presented cases, the tolerance can be described as excellent compared to other antidepressants. Although, on the whole the trial yielded very encouraging results, only double-blind trials can determine the true antidepressant efficacy of rolipram.

Influence of age, sex and osteogenesis imperfecta on count, protein content, and monoamine oxidase activity of human thrombocytes.

We determined the effect of count, age (2 to 24 year), sex, and osteogenesis imperfecta (OI) on the protein content and monoamine oxidase activity in human platelets. The reaction rate in presence of paramethoxybenzylamine was assessed in a sensitive and continuously recording spectrophotometric system. Platelets harvested from control subjects and OI patients displayed significant inverse linear correlations between count and protein content; there was near-constancy of the products of the two variables. The effects of age, sex, and osteogenesis imperfecta on protein content, count, and MAO activity were assessed by multivariate analysis of variance. It was found that, with increasing age, the count increased linearly and the protein content decreased. In patients with OI the protein content was depressed and monoamine oxidase activity elevated regardless of whether the latter was calculated on the basis of pellet protein or of count. The data suggest that, in osteogenesis imperfecta, thrombocytic monoamine and protein metabolism deviate from that of controls.

Tetrabenazine (Nitoman) therapy of chronic spontaneous oral dyskinesia. A video- and EMG-controlled study.

Therapeutic results achieved with tetrabenazine in six patients with spontaneous oral dyskinesia were evaluated by clinical rating as well as time-blind video and EMG assessment. Dramatic improvement of symptoms was observed in five of our six patients and good to satisfactory improvement in the remaining patient. The correspondence among clinical findings, EMG data, and video analysis was good. Medication was started at low doses and slowly increased. Adverse reactions (i.e., rigidity, akinesia, vasodepression) were minimal.

Strong HLA-DR expression in T cell cultures after activation is necessary for IL-2-dependent proliferation.

We have examined the appearance of DR antigens on human T cells activated by PHA, using a monoclonal anti-DR framework antibody and a large panel of human HLA typing sera. Strong DR expression within the culture by day 8 was associated with the ability of cells to grow for long periods in IL-2 containing medium, whereas weak or absent DR expression was predictive of poor in vitro growth. All the cells responded equivalently to initial stimulation with PHA. These data support the hypothesis proposed by Moretta et al. to explain blocking of IL-2-dependent proliferation by an anti-DR antibody--that DR molecules may be involved in the transmission of signals by IL-2.

[Spontaneous oral dyskinesia. Successful treatment with tetrabenazine (author's transl)]. / Spontane orale Dyskinesien. Erfolgreiche Therapie mit Tetrabenazin (Nitoman).

Tetrabenazine (50-150 mg/day) was a moderate to excellent efficacy in each of six patients with severe spontaneous oral dyskinesia. Slight akinesia, rigidity, and transient disorientation were occasional side effects that could easily be controlled and were tolerated by the patient in view of the lasting amelioration of this debilitating symptom. When tetrabenazine was administered for blepharospasm, only two of six cases showed partial improvement with more severe side effects.

Protein content and monoamine oxidase activity in platelets.

Platelets of healthy subjects and patients suffering from various disorders were assayed for their monoamine oxidase (MAO) activity and protein content. The latter tended to be lower with increasing platelet count. A linear correlation with negative slope between count and protein content was found to exist in platelets obtained from schizophrenic, parkinsonian, and (specific development) dyslexia patients. MAO activities appeared to vary significantly with respect to age and sex. In schizophrenic patients, a significant depression of MAO activity occurred which was more marked in chronic than in acute cases. Even larger activity reductions were seen in platelets of insulin-dependent diabetics while the MAO was enhanced in male dyslexic boys. When MAO activity was assessed with different substrates and methods, the results correlated well with each other. Small, but consistent discrepancies, however, arose in the schizophrenia data when compared with the control values.

Reduced monoamine oxidase activity in blood platelets from insulin-dependent diabetic subjects.

Insulin-dependent diabetics show a significant reduction in blood platelets' monoamine oxidase (MAO) activity when compared with age-matched and sex-matched controls. This does not appear to be the case for diabetics either on diet control or receiving oral hypoglycemic drugs. The important role of this enzyme in the regulation of the circulating levels of a number of monoamines known to have an inhibitory effect on insulin secretion may indicate the possible inclusion of the MAO/monoamine system in the pathophysiology of diabetes. The possibility of using platelets' MAO activity as a biologic marker for a subgroup of insulin-dependent diabetics is also discussed.

Structure activity relationships between catecholamines and the alpha-adrenergic receptor responsible for the aggregation of human platelets by epinephrine.

PEA is a potent inhibitor (Ki approximately 13 microM) of human platelet aggregation induced by epinephrine. This led us to perform an SAR study of a congeneric series of compounds in an effort to identify the molecular components of epinephrine critical to its aggregating effect upon human platelets. Phenylethanolamine was similar to PEA in inhibitory potency. However, hydroxylation of the phenyl ring diminished the inhibitory effect (Ki tyramine approximately 87 microM; Ki octopamine approximately 88 microM). Dopamine, the weakest inhibitor (Ki approximately 150 microM), was a partial agonist capable of inducing platelet aggregation in some samples of PRP. The order of potency of catecholamines as aggregating agents was epinephrine greater than norepinephrine greater than Epinine greater than dopamine. Phenylephrine, the prototype alpha-agonist, did not induce aggregation but was a potent inhibitor (Ki approximately 12 microM) of the aggregation induced by epinephrine. Isoproterenol, the prototype beta-agonist, was neither an aggregant nor an inhibitor of epinephrine-induced platelet aggregation. These findings suggest that the binding of epinephrine to the alpha-adrenergic receptor responsible for platelet aggregation is accomplished by the N-methyl amino group whereas intrinsic aggregating activity is a function of the catechol moiety.

Activation of classical complement pathway by naturally occurring heteroantibodies in normal rabbit serum. Effect on subpopulations of human lymphoid cells.

Heteroantibodies present in normal rabbit serum (NRS) are toxic to human B lymphocytes, T lymphocytes, and monocytes. Even NRS, which exhibits little back ground cytotoxicity for human lymphoid cells in conventional HLA or B-cell lymphocytotoxic assays, can be shown to contain considerable activity by making two modifications in usual procedures: by washing cells in saline or balanced salt solutions devoid of protein or sugar substances, and by increasing incubation time for 1 h to 3--4 h. Using such modifications, the cytotoxic activity of NRS towards human lymphoid cells was investigated and was found to involve activation of the classical complement pathway rather than activation of the alternate complement pathway. Residual unwanted background cytotoxicity of NRS toward human lymphoid cells can be decreased without loss of desired complement activity either by heating NRS for 15 min at 50 degrees C or by mixing NRS with small amounts of normal human serum.

Association of HLA-B8, DRw3, and anti-acetylcholine receptor antibodies in myasthenia gravis.

Twenty-eight patients with myasthenia gravis (MG), five with and 23 without thymoma, and 47 normal controls were typed for serologically defined HLA-A, B, C, and DRw antigens. Sera from all patients were titered for antibodies to acetylcholine receptors (AChR). The frequency of HLA-B8 and DRw3 in the non-thymoma MG patients was significantly higher than in the normal population. Most of the non-thymoma patients with AChR titers higher than the average level were positive for HLA-B8 and/or DRw3, while the majority of the HLA-B8(-) and/or DRw3(-) non-thymoma patients demonstrated AChR titers below average. These findings support the possibility of the existence of immune response genes in the HLA-B, DRw segment of the major histocompatibility complex which are concerned in the response to or recognition of autoantigens.

Platelet monoamine oxidase and epinephrine-induced platelet aggregation.

Platelet MAO activity and the aggregation response to epinephrine, ADP, and collagen were measured in normal subjects. There was a direct correlation between the amount of platelet MAO activity and the per cent aggregation induced by 1 and 2 micrometer epinephrine. There were lesser correlations between platelet MAO and ADP or collagen-induced aggregation. These findings suggest that platelet MAO may play a role in determining the response of human platelets to epinephrine.

Mescaline: its effects on learning rate and dopamine metabolism in goldfish (Carassius auratus).

The pharmacological action of mescaline on goldfish was studied with the Bitterman-Agranoff shock-avoidance test. In short term experiments with high mescaline doses an increase in learning rates was observed. Similar results were obtained with apomorphine and L-dopa. However, when the fish were exposed to smaller mescaline doses (or to fluphenazine) for 3 days, their ability to avoid electric shock was reduced. Apparently, mescaline induced a release of dopamine which stimulated central dopaminergic systems. Subsequently, MAO destroys the liberated dopamine. Thus, the ensuing dopamine deficit appears to be responsible for the marked changes in behavior in the chronic experiment.