Effect of embryonic 19-nortestosterone treatment and surgical bursectomy on plasma concentrations of reproductive hormones, on inhibin content in adrenals and gonads and on the histological appearance of the gonads in the young chicken.

Four-day-old chick embryos were hormonally treated with 19-nortestosterone in order to inhibit bursa development. At days 1, 4, 8, 15, 22, 29, and 36 of age, plasma, adrenals, and gonads from intact and hormonal treated chicks were collected. In embryonic nortestosterone treated males the appearance of a left 'ovotestis-like' gonad was observed. The occurrence of this ovotestis-like left gonad in the 19-nortestosterone treated male is probably a secondary effect of the in ovo treatment since surgically bursectomised chicks did not show the testicular morphology and histological changes as observed in 19-nortestosterone treated chicks. Additionally, both male and female hormonally or surgically treated chicks showed relatively enlarged adrenal glands. Hormonal bursectomy affected organ inhibin contents and plasma inhibin, testosterone, and FSH levels in males. Male hormonal treated chicks showed lower levels of plasma inhibin (p=0.0001), testosterone (p=0.01), and FSH (p=0.004), and a lower total testes inhibin content (p=0.0003) compared to intact chicks. However, none of these were significantly different between female intact and hormonal treated chicks, again indicating that the observed hormonal changes in males are not the result of the disappearance of the bursa but of the hormonal 19-NT treatment. The total adrenal inhibin content as well as the adrenal inhibin concentration were significantly higher in hormonally treated chicks than in intact chicks (p=0.0001), regardless of the sex.

Early and temporary quantitative food restriction of broiler chickens. 1. Effects on performance characteristics, mortality and meat quality.

1. An experiment was conducted with broiler chickens to determine the effects of different early food restrictions, strain (Ross 508 and Hybro G) and sex on performance, mortality carcase composition and meat characteristics. 2. Birds were restricted to 80% or 90% of ad libitum intake for 4 d (80%-4 d-group and 90%-4 d-group, respectively) or 80% for 8 d (80%-8 d-group). All restrictions started on d 4. After the periods of restriction, all birds were fed ad libitum. 3. Only the 90%-4 d-group reached a final body weight not significantly different from, but lower than, the ad libitum group. The other restrictions were too severe to allow a sufficient 'catch-up'. 4. No significant differences in food conversion and total carcase fat content between groups were observed. Abdominal fat showed a tendency to increase due to the restrictions induced. 5. There was a slight trend towards a reduced mortality and of 'sudden death syndrome' but no clear effect of food restriction on number of chickens removed with leg problems. There was no significant decrease in uniformity of the flocks due to restriction. 6. The group 80%-8 d had a significant lower yield percentage. Cut-up parts and meat quality were not changed by restriction. Ross birds had a significantly higher proportion of breast meat than Hybro chickens. Meat of female chickens seemed to be paler than that of males, possibly because of the higher proportion of carcase fat. 7. Food restriction did not always give good results. However, a mild restriction (90% for 4 d) may offer some economic advantages over an ad libitum feeding regimen, mainly by reducing mortality.

Early and temporary quantitative food restriction of broiler chickens. 2. Effects on allometric growth and growth hormone secretion.

1. An experiment was conducted with male broiler chickens to investigate the effect of different early food restriction programmes on proportional weights and allometric growth coefficients of carcase parts and on pulsatile growth hormone secretion. 2. At 4 d of age the animals were restricted to 80% or 90% of the ad libitum intake for 4 d or 80% for 8 d. When the restriction was finished, all birds received ad libitum food again. 3. On d 1, 8, 11, 14, 21, 28, 35 and 42, from each group 8 birds were weighed, stunned and killed and the weights of the stomachs, liver, heart, sternum, right thigh, right drumstick, breast and abdominal fat pad were determined. On d 29, 30, 31, 32 serial blood samples of 2 birds from each group were taken to determine the plasma growth hormone (GH) content. 4. Only the most severe restriction resulted in a tendency for the stomachs to mature earlier and the thighs to mature later. No other significant effect on the allometric coefficients could be observed. 5. It is concluded that during the restriction the birds give priority to the development of the supply organs such as the stomach, which are more important during early development at the expense of the demand tissues like breast and thigh. 6. No significant differences concerning the GH secretory parameters could be observed.

Monitoring embryo development in chicken eggs using acoustic resonance analysis.

Early embryonic development was monitored using acoustic resonance frequency analysis. Based on the course of the resonance frequency (RF) of a chicken egg during incubation, detection of viable and infertile eggs or eggs in which the embryo died during the first days of incubation was possible at the beginning of day 5. A sudden decrease of the RF of viable eggs was noticed after 96-104 h of incubation, whereas infertile eggs or eggs in which the embryo died during the first days of incubation showed a smooth, continuous increase of RF during the entire period of observation. Furthermore, the interruption of the embryonic development by cold storage after 24 or 84 h of incubation delayed the moment of decreasing RF. This proves that the sudden decrease of RF corresponds to a certain stage of development. This application of acoustic resonance frequency analysis offers the potential of early, automatic detection of inviable embryo's in eggs during incubation.

Ovarian and extraovarian sources of immunoreactive inhibin in the chicken: effects of dexamethasone.

The present study investigates whether besides the ovary, extragonadal sources contribute to the total amount of immunoreactive inhibin in the plasma of the domestic hen. A comparison of the inhibin content of different organ shows that, expressed per milligram of tissue, the adrenal ranks second only to the ovarian granulosa layer. To explore the contribution of the adrenals to plasma inhibin, dexamethasone (100 micrograms/kg BW) was injected i.v. into intact, ovariectomized, and sham-operated hens. Control animals of each experimental group were injected with saline (0.9% (w/v) NaCI). Dexamethasone significantly (P < 0.05) decreased plasma inhibin concentrations in the three groups. The suppressive effect of dexamethasone in intact hens, however, was caused by a direct effect of this synthetic glucocorticoid on the gonads. Indeed, dexamethasone decreased the production of inhibin by granulosa cells in vitro and also lowered the immunoreactive inhibin concentration in ovariectomized animals. The decreased plasma inhibin concentration in ovariectomized animals is probably due to a direct effect of dexamethasone on the adrenals. Adrenal cells produced immunoreactive inhibin in vitro. The inhibin secretion by adrenal cells was significantly (P < 0.05) depressed by dexamethasone. In conclusion, the ovary is the major source of plasma immunoreactive inhibin in the laying hen. The presence of substantial amounts of immunoreactive inhibin in the adrenal, the secretion of inhibin by cultured adrenal cells, and the decreased immunoreactive inhibin in ovariectomized animals treated with dexamethasone indicate that the adrenal is a likely source of extragonadal inhibin. The nature and the role of this adrenal inhibin remain to be investigated.

IgG antibodies to SS-B (La), RNP/Sm and DNA are produced by PWM-stimulated normal human lymphocytes in culture.

Peripheral blood lymphocytes from five healthy subjects and three patients with Sjögrens syndrome and systemic lupus erythematosus were stimulated with pokeweed mitogen to examine the effects of polyclonal activation on the secretion of autoantibodies in health and disease. Antibodies to SS-B (La), RNP/Sm and DNA were detected in supernatants from cultures from healthy controls, in some cases approaching levels secreted by the patients. All secreted autoantibodies were of IgG class and the antigen specificity of the secreted anti-SS-B was proven by cross-adsorption experiments. Our results extend the range of defined specificities of autoreactive B cells in healthy individuals. These data argue against a case for physiological deletion of autoreactive B cell clones and support theories of their active recruitment in autoimmunity.

In vitro immunoglobulin synthesis by lymphocytes from patients with rheumatoid arthritis. I. Effect of monocyte depletion and demonstration of an increased proportion of lymphocytes forming rosettes with mouse erythrocytes.

We have investigated B cell function in nine patients with rheumatoid arthritis (RA) compared to sex and age matched controls in a pokeweed mitogen driven system. Levels of IgG and IgM synthesized in the supernatant were measured by a competition ELISA. We have found that cultured mononuclear cells from RA patients showed a defective Ig synthesis when depleted of monocytes. In contrast RA mononuclear cells not depleted of monocytes produced substantial levels of Ig after stimulation by the mitogen. The percentages of T and B lymphocytes in the peripheral blood of RA patients were normal; however, an increased number of lymphocytes formed rosettes with mouse erythrocytes indicating an abnormality in the B cell pool. These results demonstrate defective in vitro immunoglobulin synthesis by RA lymphocytes and show the importance of monocytes in this culture system.

Quantitation and detection of isotypes of anti-SS-B antibodies by ELISA and Farr assays using affinity purified antigens. An approach to the investigation of Sjögren's syndrome and systemic lupus erythematosus.

Affinity purified SS-B was characterized as a protein with immunoreactive polypeptides of 40K and 29K. A modified Farr assay and an enzyme linked immunosorbent assay (ELISA) were 100- to 1,000-fold more sensitive than immunodiffusion and showed an association with the systemic manifestations of primary sicca syndrome and Sjögren's syndrome with systemic lupus erythematosus. The ELISA was sufficiently sensitive to detect class specific antibodies in saliva and lymphocyte culture supernatants.

B-lymphocyte subpopulation which forms rosettes with mouse erythrocytes increased in rheumatoid arthritis.

We studied the peripheral blood lymphocytes of 22 patients with rheumatoid arthritis (RA) for the presence of a subpopulation of cells which form rosettes with mouse erythrocytes. In normal subjects these cells have been characterised as immature B cells which are non-responsive to pokeweed mitogen. The mean percentage of mouse rosette-forming cells (MRFC) in the rheumatoid group was 13 +/- 10(mean +/- 2 SD), a significantly higher value than the control mean of 5% +/- 4% (P less than 0.001). The T- and B-cell percentages in the rheumatoid patients were normal. The ratio of MRFC: B cells derived from these results was 3:4 in RA and 1:4 in normal subjects. Pre-incubation of rheumatoid peripheral blood lymphocytes at 4 degrees C gave higher values of MRFC (19% +/- 10%) than pre-incubation at 37 degrees C (13% +/- 10%, P less than 0.02), but no such temperature effect was found in the control group. There was no correlation between MRFC and rheumatoid disease activity or the patients' drug regimens. We conclude that the threefold increase in mean MRFC in patients with rheumatoid arthritis indicates an abnormality in the circulating B-cell pool.

Transsynovial distribution of ibuprofen in arthritic patients.

Free and total ibuprofen levels in serum and synovial fluid (SF) were determined in one male and 14 female arthritic patients (mean age 56 yr, range 19 to 77) after 400 mg three times daily for 2 days. Free drug was separated by ultrafiltration and assayed by a new highly sensitive and precise gas chromatographic-mass spectrometric technique. Total protein and albumin were assayed by automatic analyzer, and the routine serum biochemical profile obtained. Serum and SF nonesterified fatty acids (NEFAs) were methylated and assayed by gas chromatography. While both total protein and albumin levels were lower (P less than 0.01) in SF, NEFA levels did not differ (p greater than 0.05). Ibuprofen readily partitions into and binds strongly to joint fluid, the total level being about half that in serum. The ratio of total ibuprofen in SF to that in serum (x = 0.41 +/- 0.17) correlated with the albumin concentration ratio (r = 0.886) indicating that albumin binding is an important determinant of ibuprofen transsynovial distribution. A low total ibuprofen SF/serum ratio, apparent for three patients on concomitant aspirin therapy, is consistent with reports of an ibuprofen-salicylate interaction. Free ibuprofen in SF (0.192 +/- 0.086 microgram ml-1) did not differ (p greater than 0.05) from serum free levels (x = 0.251 +/- 0.139 microgram ml-1), supporting the concept of the synovial cavity as a compartment readily accessible to unbound drug species. Multiple regression analyses revealed relative independence of free concentration on total levels, and the small positive and negative influences of NEFAs and albumin concentrations. Bilirubin, uric acid, cholesterol, triglycerides, and chloride ion did not correlate with serum free concentration.

Effect of histamine on the mitogenic response of human lymphocytes and its modification by cimetidine and levamisole.

The phytohaemagglutinin (PHA) mitogenic response of human blood lymphocytes demonstrated a dose-dependent inhibition by histamine (in the range 5 X 10(-7) M). Using suboptimal mitogenic doses of PHA the suppression was more pronounced. Stimulation using different doses of pokeweed mitogen (PWM) was not altered by histamine. Purified T cells and a lymphocyte population depleted of histamine-receptor-bearing cells were less sensitive to the histamine effect. In the supernatants of histamine-stimulated cultures, suppressor factor activity was found. Cimetidine could reverse the effect of histamine in a dose-dependent manner. Adding levamisole to the PHA cultures also produced some antihistaminic effect.

The inhibitory effect of D-penicillamine on human lymphocyte cultures stimulated by phytohaemagglutinin, the antagonistic action of L-cysteine and synergistic inhibition by copper sulphate.

The inhibitory effects of D-penicillamine (D-Pen) on lymphocyte activation by PHA are found to be dose-dependent, showing significant effects above a concentration of 50 microgram/ml. The cells show a diminished sensitivity to D-Pen added after 24 hours in culture, although the dose-response relationship is maintained. L-cysteine is usually found to have an insignificant effect on PHA stimulation, although enhancement and inhibition are also sometimes observed. The supplementation of culture medium with L-cysteine abolishes the inhibitory effect of D-Pen, whilst copper sulphate synergistically inhibits PHA-induced transformation. The balance between the availability of naturally occurring thiols, copper ions and administered D-Pen may be critical in determining the eventual effect of the drug on the lymphoid system. Lymphocyte suppressive soluble factors appear in the sera of patients with rheumatoid arthritis and are modified by treatment with D-Pen. Although our experiments suggest possible modes of action of the drug on the immune system as a therapeutic agent as well as a potent inducer of autoimmunity in its own right, further work is required to delineate its precise mechanism of action.