Disrupted cross-laminar cortical processing in ß amyloid pathology precedes cell death.

Disruption of neuronal networks in the Alzheimer-afflicted brain is increasingly recognized as a key correlate of cognitive and memory decline in Alzheimer patients. We hypothesized that functional synaptic disconnections within cortical columnar microcircuits by pathological ß-amyloid accumulation, rather than cell death, initially causes the cognitive impairments. During development of cortical ß-amyloidosis with still few plaques in the transgenic 5xFAD mouse model single cell resolution mapping of neuronal thallium uptake revealed that electrical activity of pyramidal cells breaks down throughout infragranular cortical layer V long before cell death occurs. Treatment of 5xFAD mice with the glutaminyl cyclase inhibitor, PQ 529, partially prevented the decline of pyramidal cell activity, indicating pyroglutamate-modified forms, potentially mixed oligomers of Aß are contributing to neuronal impairment. Laminar investigation of cortical circuit dysfunction with current source density analysis identified an early loss of excitatory synaptic input in infragranular layers, linked to pathological recurrent activations in supragranular layers. This specific disruption of normal cross-laminar cortical processing coincided with a decline of contextual fear learning.

Effects of insulin-induced hypoglycemia on beta 2-adrenoceptor density and proliferative responses of human lymphocytes.

We studied the effects of insulin (0.1 IU/kg BW, iv)-induced hypoglycemia on lymphocyte beta 2-adrenoceptor function, lymphocyte subset distribution, and proliferative response to mitogen stimulation in 10 healthy volunteers. Thirty minutes after insulin injection plasma glucose levels were markedly decreased; concomitantly, plasma epinephrine levels had increased about 10-fold; plasma norepinephrine levels, however, increased only moderately. Lymphocyte beta 2-adrenoceptor density and the cAMP response to 10 mumol/L isoproterenol stimulation were elevated; lymphocyte Ts/c-cells had increased, whereas Th-cells had decreased, resulting in a decrease in the Th-/Ts/c-cell ratio from 1.7 to 1.0. These changes were accompanied by a significantly reduced lymphocyte proliferative response (measured as [3H]thymidine uptake) to mitogen stimulation. Two hours after insulin injection plasma catecholamines, lymphocyte beta 2-adrenoceptor function, lymphocyte subset distribution, and proliferative responses had returned to nearly preinsulin levels. We conclude that acute vigorous increases in endogenous epinephrine evoked by insulin-induced hypoglycemia are associated with increases in lymphocyte beta 2-adrenoceptor function, redistribution of lymphocyte subsets, and an (at least transiently) attenuated in vitro immune responsiveness.

Resistance to insulin-induced hyperphagia in fat-preferring rats.

Outbred male Sprague-Dawley rats were screened for their macronutrient (fat, carbohydrate and protein) preference profile and divided into two groups, the low- and high fat-preferring groups each deriving 23% and 72% of its total caloric intake from fat respectively. Subcutaneous administration of bovine insulin (6U/kg) resulted in an increase in total caloric intake in the low, but not the high fat-preferring group. Furthermore, the increased caloric intake in the low fat-preferring group was entirely due to increased carbohydrate consumption. These data suggest a defect in the action of insulin in fat-preferring rats.

Regulation of dietary fat preference: establishing a reproducible profile of dietary fat preference in rats.

The mechanism(s) underlying preference for individual macronutrients (particularly fat) in diet are poorly understood. The greatest obstacle in designing experiments to define neurochemical determinants of fat preference lies in our ability to clearly identify animals' macronutrient preference (MP) profile. To this end, we have defined the role of several variables and suggested ways to design better studies to examine the mechanism of macronutrient preference. The results of these studies show that the paradigm used for MP analysis, genetics and age of the animal could clearly affect the MP profile.

Nitrite and nitrate formation on model NOx storage materials: on the influence of particle size and composition.

A well-defined model-catalyst approach has been utilized to study the formation and decomposition of nitrite and nitrate species on a model NO(x) storage material. The model system comprises BaAl(2x)O(1+3x) particles of different size and stoichiometry, prepared under ultrahigh-vacuum (UHV) conditions on Al(2)O(3)/NiAl(110). Adsorption and reaction of NO(2) has been investigated by molecular beam (MB) methods and time-resolved IR reflection absorption spectroscopy (TR-IRAS) in combination with structural characterization by scanning tunneling microscopy (STM). The growth behavior and chemical composition of the BaAl(2x)O(1+3x) particles has been investigated previously. In this work we focus on the effect of particle size and stoichiometry on the reaction with NO(2). Particles of different size and of different Ba(2+) : Al(3+) surface ion ratio are prepared by varying the preparation conditions. It is shown that at 300 K the reaction mechanism is independent of particle size and composition, involving initial nitrite formation and subsequent transformation of nitrites into surface nitrates. The coordination geometry of the surface nitrates, however, changes characteristically with particle size. For small BaAl(2x)O(1+3x) particles high temperature (800 K) oxygen treatment gives rise to particle ripening, which has a minor effect on the NO(2) uptake behavior, however. STM shows that the morphology of the particle system is largely conserved during NO(2) exposure at 300 K. The reaction is limited to the formation of surface nitrites and nitrates, which are characterized by low thermal stability and completely decompose below 500 K. As no further sintering occurs before decomposition, NO(2) uptake and release is a fully reversible process. For large BaAl(2x)O(1+3x) particles, aggregates with different Ba(2+) : Al(3+) surface ion ratio were prepared. It was shown that the stoichiometry has a major effect on the kinetics of NO(2) uptake. For barium-aluminate-like particles with high Al(3+) concentration, the formation of nitrites and nitrates on the BaAl(2x)O(1+3x) particles at 300 K is slow, and kinetically restricted to the formation of surface species. Only at elevated temperature (500 K) are surface nitrates converted into well-defined bulk Ba(NO(3))(2). This bulk Ba(NO(3))(2) exhibits substantially higher thermal stability and undergoes restructuring and sintering before it decomposes at 700 K. For Ba(2+)-rich BaAl(2x)O(1+3x) particles, on the other hand, nitrate formation occurs at a much higher rate than for the barium-aluminate-like particles. Furthermore, nitrate formation is not limited to the surface, but NO(2) exposure gives rise to the formation of amorphous bulk Ba(NO(3))(2) particles even at 300 K.

Expanded TCR Vbeta subsets of CD8(+) T-cells in late-onset myasthenia gravis: novel parallels with thymoma patients.

Little is known about pathogenesis -- and especially about involvement of CD8(+) T-cells -- in late-onset myasthenia gravis (LOMG). Remarkably, outstanding CD8(+) TCRVbeta-subset expansions were found in 64% and 72% of recent onset LOMG or thymoma-associated MG (vs. 16% with early-onset MG (p<0.0002); 21% in older controls (p<0.001)). In LOMG, ~25% of the expanded cells initially showed a naïve CD62L(+hi)/CD45RA(+) recent thymic emigrant (RTE)-like phenotype. These expansions associated significantly with IgG antibodies against cytomegalovirus (p<0.036), IL-12 and/ or IFN-alpha2 (p<0.03). The CD8(+) TCRVbeta expansions were stable over 5years, but RTE markers declined.

Recognition of monocyte-associated antigens in the dog.

Canine pregnant sera were investigated for monocyte-specific cytotoxic antibodies against a cell panel from 50 unrelated dogs. Contaminating DLA and DLB reactivity was removed by absorption on a random pool of purified lymphocytes. Two monocyte-associated antigens were recognized by two independent clusters of antisera; the existence of further antigens is suggested. In preliminary segregation studies, the inheritance of these determinants in linkage with DLA could not be observed.

Comparison between CO2 microsurgical laser and conventional meniscectomies in sheep: a preliminary report.

Temporomandibular discectomies were performed in sheep using a CO2 microsurgical laser as the experimental variable and conventional scissors and scalpels as the control. Histologic sections at 1, 3, 7, 14, and 30 days failed to reveal any significant difference between groups, nor was there any significant difference noted in the subjective evaluation of intraoperative bleeding. These findings suggest that when performing discectomies, the laser offers no advantage over conventional techniques.

Revised classification of the DLA loci by serological studies.

Peripheral blood mononuclear cells from DLA typed dogs were treated with rabbit-anti-dog-beta 2-microglobulin and subsequently with goat-anti-rabbit-immunoglobulin in order to aggregate the DLA class I molecules on the cell membrane (lysostrip). Utilizing a panel of 70 defined DLA-A and DLA-B antisera, lymphocytes treated in this way showed resistance to complement dependent lysis with monospecific DLA-A sera only, whereas reactivity of DLA-B antisera was not blocked; on the contrary, complete lympholysis with each DLA-B antiserum was recognized. Thus, the DLA-B antigens, evidently not associated with beta 2-microglobulin, are designated as candidates for class II gene products. The different reactions of DLA-C antisera after lysostrip did not allow a precise assignment of this antigen series as yet.

Animal studies on bone ingrowth kinetics of ceramic material under dynamic stress.

Two of the primary problems of implants in use today result from the materials used in their construction (metal and polyethylene) and from the necessary additional fixation with bone cement. In order to alleviate these well known difficulties, we studied ceramic material (99.7% Al2O3 with 0.25% MgO) which exhibits several advantages in these areas, but has a diminished bending strength. To take advantage of the ceramic material, we tested a cement-free implantation which should allow unhindered growth of bone tissue to or into the material. In particular, the interface relationship under load bearing was investigated. The course of the tissue differentiation and ingrowth on the surface of the ceramic implants under dynamic stress, was studied by a specifically disegned distance-spacer. These were tested on the femur of foxhounds and sheep, as well as by use of temporary immobilization. Histological investigations in intervals of 4 weeks showed the course of the development of the interface tissue. After removal of the binding materials, the behavior of the implant support is studied in 4-week intervals with free load bearing. Radiological and histological development studies are shown. The results we obtained implicate the use of ceremic impants. But the application in humans still seems to be problematic, as there is a need for designs which are loaded by pressure only.

Catecholamines increase lymphocyte beta 2-adrenergic receptors via a beta 2-adrenergic, spleen-dependent process.

We investigated the mechanisms underlying the increase in mononuclear leukocyte (MNL) beta 2-adrenergic receptor (AR) number and responsiveness after acute infusion of catecholamines. Infusion of isoproterenol and epinephrine, but not of norepinephrine, acutely increased MNL beta-AR density, and this was blocked by the beta 2-selective antagonist ICI 118,551 but not by the beta 1-selective antagonist bisoprolol, suggesting a beta 2-AR-mediated effect. Infusion of isoproterenol but not of norepinephrine also induced a lymphocytosis, with an increase in the number of circulating suppressor/cytolytic T (Ts/c)- and natural killer (NK)-cells but a decrease in helper T (Th)-cells, leading to a decreased Th-Ts/c-cell ratio. beta-AR density was higher in Ts/c-cells than in Th-cells. After isoproterenol infusion, beta-AR density was elevated in all lymphocyte subsets but not in monocytes or platelets, suggesting a lymphocyte-specific phenomenon. Infusion of isoproterenol in splenectomized patients did not alter lymphocyte subset composition and only slightly increased beta 2-AR density. In healthy subjects lymphocyte proliferation in response to various mitogens was attenuated after infusion of isoproterenol but not of norepinephrine; this effect was abolished in splenectomized patients. We conclude that the elevated MNL beta-AR density after acute exposure to beta-adrenergic agonists is caused by a release of lymphocyte subsets from the spleen into the circulation and/or by an exchange of lymphocyte subsets between the spleen and the circulation, whereby freshly released splenic lymphocytes appear to carry more beta-AR than those found in the circulation. This appears to impair immune responsiveness in a dual manner, by decreasing the Th-/Ts/c-cell ratio and by rendering lymphocytes more sensitive to the antiproliferative effects of catecholamines via a higher beta-AR density.