Studies of the mechanism of tolerance induced by short-term immunosuppression with cyclosporine in high-risk corneal allograft recipients. I. Analysis of CTL precursor frequencies

Transplantation of unmatched allogeneic corneas into highly vascularized recipient eyes under the cover of short-term immunosuppression with cyclosporine enables permanent engraftment. The aim of this study was to further elucidate the mechanism(s) underlying this tolerant state. In eight "high-risk" cornea recipients the clone sizes of donor-specific and third-party reactive cytotoxic T cell precursors were assessed by limiting dilution analyses before and at three and six months after transplantation. Acquired allograft tolerance in these patients was not accompanied by clonal reduction of donor-specific CTL-p, whereas in the case of an irreversible rejection the donor-specific CTL pool size was significantly enlarged. This donor-specific CTL-p increase could already be seen two months before clinical manifestation. These patterns differed from that of tolerant renal transplant patients, in whom marked and donor-specific reduction of CTL-p was observed. During rejection identical patterns with increasing donor-specific CTL-p frequencies were seen in both groups of patients. We conclude that induction of tolerance by short-term CsA to unmatched cornea grafts is not caused by clonal reduction of the effector precursor cell pool.     

Altered responsiveness to stress and NMDA following prenatal exposure to cocaine

Pregnant Sprague--Dawley rats were treated once daily with 40-mg/kg cocaine or saline from gestation days (GD) 12 to 21. A third group of pregnant dams was used as a pairfed control. Male and female offspring were examined for stress endurance response as determined by the cold-water swim test on postnatal days (PND) 21, 30, 40, and 60. Male and female offspring exposed to cocaine in utero were found to have diminished tolerance and altered hormonal response to stress. Moreover, prenatal cocaine exposure has been associated with significant increases in severity of N-methyl-D-aspartate (NMDA; 35 mg/kg) behavioral responses (tail twitches, wetdog shaking, and convulsion) as compared to control. Examining the experimental groups for pain sensitivity using the tail-flick and the hot-plate methods indicated that prenatal cocaine exposure altered pain sensitivity. NMDA receptor binding studies showed an increase in receptor density in the hippocampus and hypothalamus of the cocaine-treated group. These results indicate that gestational cocaine exposure is associated with long-term alterations in response to stress, NMDA receptor, and pain sensitivity in the rat offspring.     

Electrophysiologic studies on the pallido- and cerebellothalamic projections in squirrel monkeys (Saimiri sciureus)

Summary Thalamic projections of the pallidum and the deep cerebellar nuclei were studied by unitary recordings as well as field potential analysis in the thalamus of squirrel monkeys (Saimiri sciureus) under sodium pentobarbital anesthesia.Stimulation of the pallidum produced a positive field potential preceded by incoming afferent fiber volleys in the thalamus. Spontaneous discharges of thalamic neurons were suppressed during this positive potential, and intracellular recordings from the thalamic neurons revealed that the time course of this field potential corresponded to that of the hyperpolarizing potential. The hyperpolarization was presumed to be a monosynaptic inhibitory postsynaptic potential by the short synaptic delay (about 0.5–0.7 ms) and responsiveness to high frequency stimulation (over 150 Hz). The positive field potential on stimulation of the external pallidal segment was distributed in L.po (VA) and the reticular thalamic nucleus around L.po, whereas that on stimulation of the internal segment was in V.o.a (the anterior basal part of VL) and in Z.o (upper part of VL). The projection of the external segment appeared to be less dense than that of the internal segment.The projection of deep cerebellar nuclei was situated in V.o.a, V.o.p (posterior part of basal part of VL), V.o.i (VLm), the intralaminar nucleus (CL), and some part of V. im (the rostral part of VPLo). Projections of the interpositus and dentate nuclei were distributed in a more anterior part than those of the fastigial nucleus. A certain topographical arrangement of the projections of these three nuclei was found in V.o.p, V.o.i and V.im. No significant overlap was detected between projections of the pallidum and the deep cerebellar nuclei within the thalamus.     

Hormone replacement therapy and vigilance: double-blind,placebo-controlled EEG-mapping studies with an estrogen-progestogen combination (Climodien,Lafamme) versus estrogen alone in menopausal syndrome patients

The aim of the double-blind, placebo-controlled study was to investigate the effects of a continuous combined estrogen-progestogen treatment (Climodien, Lafamme) as compared with estrogen alone on vigilance in insomniac postmenopausal syndrome patients, objectified by EEG mapping. METHODS: In a 3-arm, 2-month parallel group design phase, patients received a combination of estradiol valerate 2 mg and the novel progestogen dienogest 3 mg (Climodien 2/3) or estradiol valerate 2 mg alone or placebo. In a subsequent open-label phase, all patients received estradiol valerate 2 mg+dienogest 2 mg (Climodien 2/2). EEG mapping was carried out before and after the 2-month double-blind phase as well as after the 2-month open-label treatment. RESULTS: As compared with placebo, Climodien 2/3 induced a marked and highly significant increase in absolute power in all frequency bands, specifically in alpha-2 activity. Moreover, a significant increase in relative alpha-2 power, a decrease in relative delta and beta power as well as an acceleration of the dominant frequency and of the delta and alpha centroids suggested a marked improvement in vigilance. In contrast, under estradiol valerate 2 mg alone, only a slight augmentation of alpha and attenuation of relative delta and beta power occurred, suggesting only a slight vigilance improvement as compared with placebo. Thus, dienogest 2 mg increased the estrogen effect, which was also confirmed by a statistical evaluation of the differences between Climodien 2/3 and estradiol valerate alone (augmentation of alpha-2, attenuation of relative beta, acceleration of the dominant frequency). Moreover, Climodien 2/2 also markedly increased alpha-2 power, decreased relative beta-2 power and accelerated the alpha centroid. Finally, comparing Climodien 2/3 with Climodien 2/2, there was even a dose-efficacy relation. CONCLUSIONS: Estradiol valerate 2 mg improves vigilance slightly, thereby confirming previous findings. The additional administration of dienogest does not minimize the effect of estrogen, but on the contrary increases it, which makes the combination superior to both placebo and estradiol valerate alone. Vigilance improvement may be of great therapeutic benefit to menopausal syndrome patients at a time when increased adaptability is needed to adjust to increasing sexual, marital, occupational and social difficulties known to occur specifically in this period of life.     

Bicycle helmet ventilation and comfort angle dependence

Five modern bicycle helmets were studied to elucidate some of the variations in ventilation performance, using both a heated manikin headform and human subjects (n=7). Wind speed and head angle were varied to test their influence on the measured steady-state heat exchange (cooling power) in the skull section of the headform. The cooling power transmitted by the helmets varied from about 60% to over 90% of that of the nude headform, illustrating the range of present manufacturer designs. Angling the head forward by 30° was found to provide better cooling power to the skull (up to 25%) for three of the helmets and almost equal cooling power in the remaining two cases. Comparisons of skull ventilation at these angles with human subjects strongly supported the headform results.     

A duplication/paracentric inversion associated with familial X-linked deafness (DFN3) suggests the presence of a regulatory element more than 400 kb upstream of the POU3F4 gene

X-linked deafness with stapes fixation (DFN3) is caused by mutationsin the POU3F4 gene at Xq21.1. By employing pulsed field gelelectrophoresis (PFGE) we identified a chromosomal aberrationin the DNA of a DFN3 patient who did not show alterations inthe open reading frame (ORF) of POU3F4. Southern blot analysisindicated that a DNA segment of 150 kb, located 170 kb proximalto the POU3F4 gene, was duplicated. Fluorescence in situ hybridization(FISH) analysis, PFGE, and detailed Southern analysis revealedthat this duplication is part of a more complex rearrangementincluding a paracentric inversion involving the Xq21.1 region,and presumably the Xq21.3 region. Since at least two DFN3-associatedminideletions are situated proximal to the duplicated segment,the inversion most likely disconnects the POU3F4 gene from aregulatory element which is located at a distance of at least400 kb upstream of the POU3F4 gene.     

Reactivity of microhemagglutination, fluorescent treponemal antibody absorption, Venereal Disease Research Laboratory, and rapid plasma reagin tests in primary syphilis.

Seroreactivity of sera from 109 patients with first-infection primary syphilis was 98.2% in the fluorescent treponemal antibody absorption test, 92.7% in the rapid plasma reagin 18-mm circle card test, 72.5% in the microhemagglutination test (MHA-TP), and 72.5% in the Venereal Disease Research Laboratory test. Seroreactivity of sera from 18 patients with primary syphilis with documented previous infection(s) was 100% in the fluorescent treponemal antibody absorption test, the rapid plasma reagin 18-mm circle card test, and the MHA-TP test and 88.9% in the Venereal Disease Research Laboratory test. The MHA-TP test failed to confirm reactivity in 13 of 79 sera which were reactive in the Venereal Disease Research Laboratory test and in 24 of 101 sera which were reactive in the rapid plasma reagin 18-mm circle card test. Testing another production lot of MHA-TP reagents resulted in even poorer correlation. The reactivity of the MHA-TP test in primary syphilis appeared to vary with the sensitivity of the production lot of reagents.     

Cell-mediated immune responses between HLA-identical siblings: recognition of antigenic changes associated with acute myelogenous leukaemia.

Cell-mediated immune reactions between a patient suffering from acute myelogenous leukaemia (AML) and an HLA-identical sibling were studied in order to characterize the in vitro reactions in MLC and CML prior to bone marrow transplantation. Our results indicated that antigenic differences were detectable between the blasts and the remission lymphocytes. While the normal sibling did not respond in MLC to her HLA-identical sister's remission lymphocytes, there was an anti-blast response. This proliferative response, however, did not lead to the development of detectable cytotoxic cells capable of destroying blast cells. Unrelated individuals, on the other hand, responded strongly both in MLC and CML to the allogeneic tumour blasts and remission lymphocytes of the patient and the lymphocytes of the healthy sibling. The kinetics and magnitude of the MLC response to blast cells was different from that to remission lymphocytes. This response indicated that the blast cells expressed antigenic differences which were recognized in MLC by both the HLA-identical sibling and unrelated individuals. Furthermore, these tumour cells were capable of sensitizing allogeneic, but not syngeneic lymphocytes to become cytotoxic, though they seemed to be more resistant to destruction in CML than normal cells.     

A simple rosette assay for demonstration of complement receptor sites using complement-coated zymosan beads.

In the present article we describe a simple rosette assay for detection of C3 receptor-bearing B lymphocytes with complement (C)-coated zymosan (Zy) beads. Zy coated with murine or human C bound to a distinct population of human and mouse lymphocytes as well as to the majority of lymphoblastoid cells of several human established cell lines. Rosette formation was also observed with human red blood cells, with human monocytes and neutrophils. Experiments with anti-immunoglobulin sera, with other B and T cell markers and with mouse thymocytes proved that the capacity to bind C-coated Zy is primarily a feature of B lymphocytes. The following findings suggested that C-coated Zy is bound via receptor sites for the activated components of C3: (a) Zy coated with C3-deficient serum failed to bind, (b) comparable percentages of various lymphoid and nonlymphoid cells formed rosettes with C-coated Zy as well as with antibody and C-coated sheep red blood cells, and (c) antibodies against human or murine C3 inhibited the binding of C-coated Zy.