Neonatal β-endorphin and sexual behavior

β-Endorphin (β-END) has been found to decrease copulatory behavior in male rats (Meyerson & Terenius 1977). This effect appears within 10 min, lasts for about an hour and is also exerted after castration with subsequent testosterone substitution (Meyerson 1981). In addition, β-END is known to influence neuroendocrine functions (for ref., see van Vugt & Meites 1980). Observations hitherto suggest that β-END influences copulatory behavior rather by a direct central nervous action than as a secondary effect to altered endocrine secretion.     

β1-and β2-adrenoceptor-mediated secretion of amylase from incubated rat parotid gland

The present in vitro investigation was undertaken in an attempt to obtain further information on β-adrenoceptor specificity and action in the rat parotid gland, with regard to amylase secretion. The β₁-selective agonist prenalterol was roughly 800 times more potent than the β₂-agonist terbutaline, and about 5 times more effective than noradrenaline in evoking amylase release. Propranolol was the most effective inhibitor of amylase release in all experiments. The β₁-selective antagonist metoprolol and H104/08 were also effective blockers of maximal noradrenaline-and prenalterol-induced release. The inhibition curves displayed biphasic shapes when amylase secretion was induced by noradrenaline, but not when prenalterol was the secretagogue. The β₂-antagonist H35/25 was without effect on maximal noradrenaline-and prenalterol-stimulated secretion. The amylase release evoked by submaximal concentration of terbutaline was inhibited by the two antagonists H35/25 and IPS 339. In another series of experiments propranolol and metoprolol clearly shifted the noradrenaline concentration-response curve to the right, whereas H35/25 was without effect. The results further demonstrate the major importance of the β₁-adrenoceptor (noradrenaline-activated) in eliciting amylase release from the rat parotid gland. However, it is also suggested that the β₂-adrenoceptors (terbutaline-activated) may to some extent serve the same function.     

Tenuifolin,an extract derived from tenuigenin,inhibits amyloid‐β secretion in vitro

Aim: Previous studies have shown that tenuigenin, a crude extract of Polygala tenuifolia Willd. that is commonly used in traditional Chinese herbal medicine for memory loss, can reduce the secretion of Aβ from cultured cells. However, the mechanism underlying this effect and the active compound derived from tenuigenin is unknown. In this study, a purified component of tenuigenin, tenuifolin, was examined and revealed to be an effective compound in vitro. Methods: Aβ secretion from three sets of COS‐7 cells, each carrying a plasmid expressing a different form of APP was examined following the treatment with tenuifolin. Initially, tenuifolin was determined to have no inherent toxicity to either the transfected or wild type cells at the effective concentrations. Cells were then treated with 0.5–2.0 μg mL^?1 tenuifolin for 12 h and their media were examined via an ELISA for Aβ1‐40 and Aβ‐42. Results: We found that treatment with 2.0 μg mL^?1 tenuifolin significantly decreased Aβ secretion from COS‐7 cells without altering the ratio of Aβ1‐40 and Aβ‐42. This effect is most probably due to inhibition of the β‐site APP cleaving enzyme as Aβ secretion was not inhibited from cells expressing the C99 fragment. Conclusion: Tenuifolin is an effective compound from tenuigenin. We believe that this finding should lead the way for future experiments to determine the exact mechanism for tenuifolin’s effect on Aβ secretion.     

Immunoreactive β-endorphin and acth in the same neurons of the hypothalamic arcuate nucleus in the rat

Immunoreactive ACTH and β-endorphin (β-End) were localized in the brain and pituitaries of normal and colchicine-treated rats, using the immunoperoxidase method at the light microscopic level. On adjacent serial 5-μm paraffin sections of anterior pituitaries, both ACTH and β-End could be found in the same cells. On adjacent 5-μm paraffin sections of brains of colchicine-treated rats, both ACTH and β-End could be found in the same perikarya of hypothalamic arcuate nucleus neurons. It appeared that all perikarya containing β-End contained ACTH as well, suggesting that neurons producing β-End also produce ACTH. Pathways of ACTH fibers corresponded to pathways of β-End fibers. These findings suggest that the synthesis, and transport, of ACTH and β-End are linked in the brain as well as in the pituitary, possibly through a common precursor.     

γδ T cells are secondary participants in acute graft-versus-host reactions initiated by CD4+ αβT cells

To examine the role of T cell subpopulations in an acute graft-versus-host (GVH) reaction, γδ T cells and αβ T cells expressing one of the two prototypic Vβ gene families were negatively isolated from adult blood samples and injected into allogeneic chick embryos. CD4⁺ αβ T cells expressing either Vβ1 or Vβ2 receptors were equally capable of inducing acute GVH reactions, consistent with the idea that αβ T cell alloreactivity is determined by CDR3 variability. By themselves, the γδ T cells were incapable of inducing GVH reactions. However, host γδ T cells were recruited into the donor αβ T cell-initiated lesions, where they were activated and induced to proliferate. The data suggest that γβ T cells may play a secondary role in GVH reactions.     

Immunoreactive β-endorphin in the plasma, pituitary and hypothalamus of young and old male rats

Immunoreactive beta-endorphin (IR-β-ENDO) was measured in the plasma, pituitary, and hypothalamus of young (3–5 mo.) and old (19–23 mo.) male Sprague-Dawley rats, using a specific radioimmunoassay. Plasma IR-β-ENDO in old male rats (3.44±10.54 ng/ml) was more than three times higher than values observed in young male rats (1.00±0.10 ng/ml). Pituitary content and concentration of IR-β-ENDO also were significantly greater in the old (5.85±0.51 μg/gland and 1.17±0.10 μg/mg protein) than in the young (3.53±0.29 μg/gland and 0.78±0.06 μg/mg protein) male rats. The content of IR-β-ENDO in the hypothalamus of old and young rats was nearly the same (43.45±2.47 and 49.88±6.35 ng/hypothalamus, respectively), whereas the concentration of IR-β-ENDO in the hypothalamus of the old male rats (3.89±0.25 ng/mg protein) was approximately 50% lower than that observed in the young male rats (7.80±0.85 ng/mg protein). These changes in plasma, pituitary, and hypothalamic IR-β-ENDO may contribute to the increase in prolactin and decrease in gonadotropins observed in old male rats, since β-ENDO administration is known to produce these effects on prolactin and gonadotropin secretion.     

Role of the thymus in the generation of skin-homing αβ and γδ virgin T cells

Current models of lymphocyte traffic suggest that homing specificities of T cells to tissues such as skin are generated outside the thymus as a result of activation of naive T cells by antigen in lymph nodes. Virgin T cells are thought to home to high endothelial venules in lymph nodes, but are thought to be unable to home to extra-lymphoid tissues such as skin. We used the technique of in situ labeling of the thymus with fluorescein isothiocyanate to examine the homing specificities of authentically naive T cells in vivo, immediately after their export from the thymus. We report that homing specificities for skin as well as lymph node are imprinted on T cells inside the thymus, independent of antigen. We also show that both αβ and γδ emigrant T cells exhibit homing patterns to skin and lymph nodes which are identical to those of mature T cells. Our findings demonstrate a key role for the thymus in the induction of skin-homing specificities on T cells indicating that skin-homing specificities of T cells are not generated solely outside the thymus as a result of the activation of virgin T cells by antigen. The migration of thymic emigrants to extra-lymphoid tissues within a few hours of leaving the thymus may have implications for mechanisms of peripheral self-tolerance. This pathway provides an opportunity for direct virgin T cell interactions with self components only expressed in the periphery at a time when emigrants may be more susceptible to tolerance induction than mature circulating T cells.     

Immunoreactive β-endorphin in the plasma, pituitary and hypothalamus of young and old male rats

Immunoreactive beta-endorphin (IR-β-ENDO) was measured in the plasma, pituitary, and hypothalamus of young (3–5 mo.) and old (19–23 mo.) male Sprague-Dawley rats, using a specific radioimmunoassay. Plasma IR-β-ENDO in old male rats (3.44±10.54 ng/ml) was more than three times higher than values observed in young male rats (1.00±0.10 ng/ml). Pituitary content and concentration of IR-β-ENDO also were significantly greater in the old (5.85±0.51 μg/gland and 1.17±0.10 μg/mg protein) than in the young (3.53±0.29 μg/gland and 0.78±0.06 μg/mg protein) male rats. The content of IR-β-ENDO in the hypothalamus of old and young rats was nearly the same (43.45±2.47 and 49.88±6.35 ng/hypothalamus, respectively), whereas the concentration of IR-β-ENDO in the hypothalamus of the old male rats (3.89±0.25 ng/mg protein) was approximately 50% lower than that observed in the young male rats (7.80±0.85 ng/mg protein). These changes in plasma, pituitary, and hypothalamic IR-β-ENDO may contribute to the increase in prolactin and decrease in gonadotropins observed in old male rats, since β-ENDO administration is known to produce these effects on prolactin and gonadotropin secretion.     

Patterns of lymphokine secretion amongst mouse γδ T cell clones

Although the patterns of lymphokine (LK) secretion by CD4 and CD8 αβ T cells have been extensively studied, the question of whether γδ T cells display patterns of restricted LK production and whether these patterns are the same as seen in conventional αδ T cells has not been previously addressed. In this study we generated panels of γδ T cell clones from normal C57BL/6 and BALB/c mice using a lectin-driven system and compared their patterns of secretion of nine LK with those of CD4 and CD8 αβ T cell clones generated in the same system. The results showed that γδ T cell clones displayed nonrandom patterns of highly restricted LK production with a strong bias towards the production of type 1 LK. The dominant pattern was one of high level secretion of interferon-γ and tumor necrosis factor (TNF), with variable production of interleukin (IL)-2, and little or none of the type 2 LK IL-4, IL-5, IL-6, and IL-10. This pattern differed significantly from that of CD4 Th1 clones in that γδ clones showed a striking deficiency in the production of IL-3 and granulocyte/macrophage colony-stimulating factor. A small subset of γδ clones displayed a novel pattern, in which the only LK produced in substantial quantity were TNF and variable amounts of IL-2. The bias of γδ T cells towards type 1 LK production was not an artefact associated with cloning because bulk populations of splenic γδ T cells behaved in the same way, even when activated in the presence of high concentrations of IL-4.     

CD45RA expression on γδ and αβ T cells emigrating from the fetal and postnatal thymus

We have compared the expression of CD45RA on αβ and γδ T cells emigrating from the fetal and postnatal thymus. The fetal and postnatal thymus export both CD45RA⁺ and CD45RA^- T cells. The number of γδ⁺CD45RA⁺ T cells was remarkably constant regardless of stage of ontogeny or T cell maturity. Around 5--8% of γδ thymic emigrants, thymocytes and peripheral blood lymphocytes expressed CD45RA in both fetal and postnatal animals. In contrast to γδ T cells, up to one quarter of both fetal and postnatal αβ emigrants expressed CD45RA. Post-thymic maturation of CD45RA expression on αβ emigrants, which occurred both before and after birth, appeared to be antigen independent.     

αβ Lineage-committed thymocytes can be rescued by the γδ T cell receptor (TCR) in the absence of TCR β chain

Commitment of the αβ and γδ T cell lineages within the thymus has been studied in T cell receptor (TCR)-transgenic and TCR mutant murine strains. TCRγδ-transgenic or TCRβ knockout mice, both of which are unable to generate TCRαβ-positive T cells, develop phenotypically αβ-like thymocytes in significant proportions. We provide evidence that in the absence of functional TCRβ protein, the γδTCR can promote the development of αβ-like thymocytes, which, however, do not expand significantly and do not mature into γδ T cells. These results show that commitment to the αβ lineage can be determined independently of the isotype of the TCR, and suggest that αβ versus γδ T cell lineage commitment is principally regulated by mechanisms distinct from TCR-mediated selection. To accommodate our data and those reported previously on the effect of TCRγ and δ gene rearrangements on αβ T cell development, we propose a model in which lineage commitment occurs independently of TCR gene rearrangement.     

Effect of stress and dexamethasone on immunoreactive β-endorphin levels in rat hypothalamus and pineal

In response to mild stress the levels of immunoreactive β-endorphin in rat anterior pituitary, hypothalamus and pineal fell within 10 minutes from 210 to 129 pmol/lobe, 1.47 to 0.89 pmol/mg protein and 2.53 to 0.41 pmol/gland, respectively. No alterations were found to take place in β-endorphin levels in posterior pituitary or plasma. Dexamethasone pre-treatment given 18 h prior to stress resulted in significantly greater reduction of β-endorphin levels in hypothalamus and pineal than stress alone—hypothalamic levels fell to 0.73 pmol/mg protein and pineal to 0.07 pmol/gland. Plasma β-endorphin levels in dexamethasone pretreated stressed rats were significantly lower than in intact rats (42 fmol/ml vs. 98 fmol/ml). The almost complete disappearance of β-endorphin from the pineal in response to stress and dexamethasone suggests that pineal does not itself synthesize the hormone but only utilizes and/or stores it. Gel filtration analysis of the β-endorphin im-munoreactivity in tissue extracts and plasma showed that anterior pituitary and plasma contain three immunoreactive components, eluting like β-endorphin,βP-Epotropin and pro-opiocortin, whereas only β-endorphin-like material was detected in posterior pituitary, hypothalamus and pineal.     

Developmental expression of the αIELβ7 integrin on T cell receptor γδ and T cell receptor αβ T cells

A novel monoclonal antibody, 2E7, was shown by immunoprecipitation to be reactive with the α_IELβ7 integrin and was employed to analyze the expression of this integrin in lymphocyte subsets and during T cell ontogeny. In adult lymph nodes, α_IEL was expressed at low levels by 40–70% of CD8⁺ T cells and < 5% of CD4⁺ T cells. However, virtually all intestinal intraepithelial lymphocytes and ?20% of lamina propria CD4⁺ T cells were 2E7⁺, indicating a preferential expression of this integrin on mucosal T cells. Examination of α_IEL integrin expression during thymus ontogeny revealed that ?3–5% of fetal or adult thymocytes were 2E7⁺. Interestingly, early in fetal thymus ontogeny, ?40% of 2E7⁺ cells expressed T cell receptor (TcR)-γδ and this subset persisted through birth. A developmental switch occurred such that 2E7⁺ TcR^? CD4^?8⁺ cells detected on fetal day 19 were followed by 2E7⁺ TcR-αβ CD4^?8⁺ cells in the neonatal thymus. The latter population persisted throughout thymus ontogeny into adulthood. Interestingly, a subset of TcR-γδ Vγ3⁺ day 16 fetal thymocyte dendritic epidermal cell (DEC) precursors were 2E7⁺, but all mature DEC expressed high levels of α_IEL integrin, suggesting that the α_IEL integrin was acquired late in DEC maturation. This possibility was strenghthened by immunohistochemical localization of the majority of 2E7⁺ γδ and αβ T cells to the medullary regions of the thymus. Overall, the results demonstrate a developmentally ordered expression pattern of the α_IELβ₇ integrin that suggests a common function for this integrin during TcR-γδ and -αβ CD4^?8⁺ T cell thymocyte development or perhaps in effector functions for these subsets.     

Effects of α- and β-adrenoceptor stimulation on 45Ca2+ efflux and insulin secretion from perfused rat islets

Exposure to the β₂-adrenoceptor agonist terbutaline resulted in a transient stimulation of ⁴⁵Ca²⁺ efflux from ⁴⁵Ca²⁺ preloaded rat islets perfused in 2 mm Ca²⁺ and 8.3 mm glucose. Concomitantly, an increase in insulin secretion occurred. Under the same experimental conditions, the α-adrenoceptor agonist noradrenaline promptly inhibited insulin release without any apparent influence on ⁴⁵Ca²⁺ efflux. In contrast, in a medium containing 2 mm Ca²⁺ and a low glucose concentration (2.8 mm), terbutaline stimulated insulin secretion without any apparent effects on ⁴⁵Ca²⁺ efflux. Noradrenaline had no effect on insulin secretion or ⁴⁵Ca²⁺ efflux in this medium. When islets were perfused with 8.3 mm glucose in a Ca²⁺ deficient medium, with or without addition of the chelating agent EGTA, terbutaline induced a marginal stimulation of insulin secretion and a negligible stimulation of ⁴⁵Ca²⁺ efflux. On the contrary, noradrenaline stimulated to an immediate and notable ⁴⁵Ca²⁺ efflux in these Ca²⁺ deficient media. Noradrenaline also clearly inhibited insulin secretion, though less markedly and with a slower onset than in islets perfused in 2 mm Ca²⁺. When the islets were perfused in a Ca²⁺ deficient medium with 2.8 mm glucose, terbutaline had a slight insulin releasing effect, but stimulated ⁴⁵Ca²⁺ efflux potently. Noradrenaline had no influence on insulin secretion but a weak stimulatory effect on ⁴⁵Ca²⁺ efflux. The data suggest that the β₂-adrenoceptor agonist terbutaline has the ability to stimulate insulin secretion in perfused rat islets, requiring extracellular Ca²⁺ for the full expression of its effects. These effects may be exerted through a Ca²⁺-Ca²⁺ exchange over the cell membrane and/or through cAMP and intracellular Ca²⁺ perturbations. Moreover, terbutaline directly stimulates ⁴⁵Ca²⁺ efflux, an effect inhibited by glucose. Further, the α-adrenoceptor agonist noradrenaline can inhibit insulin secretion in the absence of extracellular Ca²⁺, but the full expression of its inhibitory action is dependent on extracellular Ca²⁺ and glucose. In addition, noradrenaline stimulates ⁴⁵Ca²⁺ efflux in a Ca²⁺ deficient medium, an effect which appears independent of the glucose concentration.     

In vivo and in vitro studies on effects of β-endorphin and naloxone on sex steroids in the water frog,Rana esculenta

The effects of β-endorphin and its receptor antagonist, naloxone, on progesterone, androgens, and oestradiol-17β release in male and female Rana esculenta were studied in vivo and in vitro. In the in vivo experiments the frogs underwent hypophysectomy, gonadectomy or both, or were left intact; the animals were injected with β-endorphin or naloxone and killed after 15, 30, 90 and 240 min. In the in vitro experiments inter-renal, testis and ovary, all with and without added pituitary, were incubated with β-endorphin or naloxone for 10, 20, 40 and 80 min. The in vivo and in vitro data from males and females were in agreement. In vivo β-endorphin increased progesterone in all experimental groups and oestradiol in intact and hypophysectomized frogs, while it decreased androgens in all experimental groups. In vitro β-endorphin increased progesterone in inter-renal and gonadal tissue, and oestradiol in gonads only, while it decreased androgens in inter-renals and gonads. In vivo and in vitro naloxone induced opposite effects to β-endorphin. These data suggest that in Rana esculenta, opioids are involved in the modulation of hypothalamo-pituitary-inter-renal and gonadal axes. In particular, the data indicate a direct effect of opioids on inter-renal and gonadal sex steroid production.     

Copolymerization of γ-butyrolactone and β-butyrolactone

The copolymerization of a 4-membered β-butyrolactone (βBL) and a thermodynamically stable 5-membered γ-butyrolactone (γBL) proceeds in the bulk state with BF₃ · OEt₂ as a catalyst at room temperature to give poly[(3-hydroxybutyrate)-co-(4-hydroxybutyrate)] (P(3HB-co-4HB)) whose structure is identical with that of a polyester formed by microorganisms. The copolymer structure was confirmed by ¹H and ¹³C NMR spectroscopy. The monomer reactivity ratios were r_(γBL) = 0.48 and r_(βBL) = 0.58, respectively, and the unit composition of 4HB increases to 56% at high γBL to βBL ratio in the feed. End group analysis of the copolymer suggests the presence of a hydroxyl group and a carboxyl group at the ends of each polymer molecule. It was postulated that the monomers activated by BF₃ react with the hydroxy group derived from the water contaminant in the monomers.     

Functional β1 - and β2-adrenoceptors in the human myocardium

Functional β-adrenoceptor populations in the human heart were studied in vitro in electrically-paced strips of the right auricular and ventricular myocardium. The relative potency of selected agonists in producing inotropic responses (T_max, T′_max) in the presence of blockers for neuronal and extraneuronal uptakes was found to be as follows: isoprenaline > noradrenaline = adrenaline = salbutamol > dobutamine. Prenalterol had a negative inotropic effect in these preparations. The selective β₁ -(practoloI) and β₂-(H 35/25) blockers reduced inotropic responses to adrenaline (T_max, T′_max) and noradrenaline (T′_max) in the auricular strips. These results indicate the participation of β₂-adrenoceptors in inotropic responses in the human auricular and ventricular myocardium. For comparison, inotropic responses of electrically-paced rat myocardium to β-adrenergic agonists in the presence of blockers for neuronal and extraneuronal uptakes were likewise studied. The relative potencies for T_max were: noradrenaline = adrenaline > prenalterol > dobutamine = salbutamol. Given the high relative potency of salbutamol in the human myocardial strips (analogous to that previous shown in the β₂-dominated atria of the frog and trout) and the low relative potency of salbutamol in the rat tissue, these findings indicate a greater population of functionally active β₂-adrenoceptors in the human than in the rat myocardium.     

Transendothelial chemotaxis of human αβ and γδ T lymphocytes to chemokines

Two subpopulations of human T lymphocytes expressing different antigen receptors, α / β and γ / δ, emigrate into inflamed tissues in distinctive patterns. We compared the transmigration of α / β and γ / δ T cells to C-C and C-X-C chemokines using an in vitro transendothelial chemotaxis assay. The C-C chemokines monocyte chemoattractant protein (MCP)-1, RANTES, macrophage inflammatory protein (MIP)-1α and MIP-1β stimulated similar, dose-dependent chemotaxis of purified γ / δ T cells, whereas MCP-1, RANTES, and MIP-1α pro duced greater chemotaxis of purified α / β T cells than MIP-1β. In contrast, the C-X-C chemokines interleukin (IL)-8 and interferon-γ inducible protein-10 (IP-10) did not promote chemotaxis of either α / β or γ / δ T cells. Three γ / δ T cell clones with differing CD4 and CD8 phenotypes also migrated exclusively to C-C chemokines. Phenotypic analysis of mononuclear cells that transmigrated from an input population of unfractionated peripheral blood mono nuclear cells confirmed the results with purified γ / δ T cells. Our data demonstrate that human peripheral blood α / β and γ / δ T cells can transmigrate to MCP-1, RANTES, MIP-1α, and MIP-1β, and suggest that both T lymphocyte subpopulations share the capacity to emigrate in response to C-C chemokines during inflammation.     

Human β‐defensin 3 has immunosuppressive activity in vitro and in vivo

β‐defensins are antimicrobial peptides with an essential role in the innate immune response. In addition β‐defensins can also chemoattract cells involved in adaptive immunity. Until now, based on evidence from dendritic cell stimulation, human β defensin‐3 (hBD3) was considered pro‐inflammatory. We present evidence here that hBD3 lacks pro‐inflammatory activity in human and mouse primary M?. In addition, in the presence of LPS, hBD3 and the murine orthologue Defb14 (but not hBD2), effectively inhibit TNF‐α and IL‐6 accumulation implying an anti‐inflammatory function. hBD3 also inhibits CD40/IFN‐γ stimulation of M? and in vivo, hBD3 significantly reduces the LPS‐induced TNF‐α level in serum. Recent work has revealed that hBD3 binds melanocortin receptors but we provide evidence that these are not involved in hBD3 immunomodulatory activity. This implies a dual role for hBD3 in antimicrobial activity and resolution of inflammation.