Excess type I interferon signaling in the mouse seminiferous tubules leads to germ cell loss and sterility

Type I (α and β) interferons (IFNs) elicit antiproliferative and antiviral activities via the surface receptor IFNAR. Serendipitous observations in transgenic mice in 1988 strongly suggested that IFNα/β overexpression in the testis disrupts spermatogenesis. Here, we compare a new mouse strain transgenic for IFNβ (Tg10) and a sister strain lacking the IFNAR1 subunit of IFNAR (Tg10-Ifnar1(-/-)), both strains expressing the transgene in the testis. The main source of IFNβ RNA was the spermatid population. Importantly, the Tg10 mice, but not the double mutant Tg10-Ifnar1(-/-), showed altered spermatogenesis. The first IFNAR-dependent histological alteration was a higher apoptosis index in all germ cell categories apart from non-dividing spermatogonia. This occurred 3 weeks after the onset of IFNβ production at postnatal day 20 and in the absence of somatic cell defects in terms of cell number, expression of specific cell markers, and hormonal activities. Several known interferon-stimulated genes were up-regulated in Tg10 Sertoli cells and prepachytene germ cells but not in pachytene spermatocytes and spermatids. In concordance with this, pachytene spermatocytes and spermatids isolated from wild-type testes did not display measurable amounts of IFNAR1 and phosphorylated STAT1 upon IFNβ challenge in vitro, suggesting hyporesponsiveness of these cell types to IFN. At day 60, Tg10 males were sterile, and Sertoli cells showed increased amounts of anti-Mullerian hormone and decreased production of inhibin B, both probably attributable to the massive germ cell loss. Type I interferon signaling may lead to idiopathic infertilities by affecting the interplay between germ cells and Sertoli cells.     

Endocytosis promotes rapid dopaminergic signaling

D(1) dopamine receptors are primary mediators of dopaminergic signaling in the CNS. These receptors internalize rapidly following agonist-induced activation, but the functional significance of this process is unknown. We investigated D(1) receptor endocytosis and signaling in HEK293 cells and cultured striatal neurons using real-time fluorescence imaging and cAMP biosensor technology. Agonist-induced activation of D(1) receptors promoted endocytosis of receptors with a time course overlapping that of acute cAMP accumulation. Inhibiting receptor endocytosis blunted acute D(1) receptor-mediated signaling in both dissociated cells and striatal slice preparations. Although endocytic inhibition markedly attenuated acute cAMP accumulation, inhibiting the subsequent recycling of receptors had no effect. Further, D(1) receptors localized in close proximity to endomembrane-associated trimeric G protein and adenylyl cyclase immediately after endocytosis. Together, these results suggest a previously unanticipated role of endocytosis, and the early endocytic pathway, in supporting rapid dopaminergic neurotransmission.     

Comparison of physicochemical properties and effects of heating regimes on stored Apis mellifera and Apis florea honey

Many of the components, which render honey its specific aroma, flavor, and biological activity, are unstable over time and thermolabile. This study was aimed to compare the chemical composition, effect of heating as well as the time of heat exposure, and storage period on the quality of honey samples from Apis mellifera (A.m.) and Apis florea (A.f.). Methods of the Association of the Official Analytical Chemists (AOAC) were used in this study. The mean values for both A.m. and A.f. honeys were, respectively: moisture (18.5, 13.7%); glucose (35.2, 36.3%); fructose (33.7, 33.8%); sucrose (7.3, 2.9%); invert sugar (68.9, 70.4%); ash (0.26, 1.1%); acidity (51.8, 98.4 meq/kg); pH (3.6, 4.4) and Hydroxy methyl furfural (HMF) (3.78, 3.17 mg/100 g). Honey from A. florea contained less moisture, have higher acidity and ash contents than A. mellifera honey. Significant alterations (P < 0.05) in glucose, fructose, sucrose, and acidity were noticed after six months. Honeys exposed to heating for 15 and 30 min at 50 and 80 °C have shown increased thermo-generated HMF after 15, 30, and 45 days. HMF reached 16.30 ± 1.1 in A. mellifera and 7.41 ± 1.4 mg/100 g in A. florea honeys that exposed for 30 min at 80 °C. Honey from A. florea showed more heat tolerance to thermo-generation of HMF than honey from A. mellifera.     

Enhanced resistance to HIV-1 replication in U937 cells stably transfected with the human IFN-beta gene behind an MHC promoter fragment

Cells of the monocyte lineage act as a major reservoir for HIV, and ways of enhancing the resistance of mononuclear phagocytes to HIV replication would be useful for delaying the onset of AIDS in infected individuals. Seif et al. (J. Virol. 65:664, 1991) have recently shown the possibility of obtaining stable antiviral expression (SAVE), directed against three nonretroviral RNA viruses, and normal cell viability in a significant percentage of murine BALB/c 3T3 cells transformed with an IFN-beta expression plasmid under the control of the 0.6-kb XhoII-NruI promoter region of the murine H-2Kb MHC gene. In the present paper, we show that it is possible to establish SAVE in human promonocytic cells. Cells of the human promonocytic U937 line were stably transfected with a human IFN-beta expression plasmid carrying the neo- and human IFN-beta-coding sequences under the control of the H-2Kb promoter fragment previously used in murine cells. After selection with G418, two transformed clones were isolated that released small amounts of human IFN-beta into the culture medium, without affecting the expression of CD4 and leucocyte function-associated Ag-1 differentiation Ag. The presence of construct-derived IFN-beta mRNA was demonstrated by polymerase chain reaction amplification of cDNA, and the level of 2-5A synthetase, one of the major IFN-induced antiviral proteins, was shown to be constitutively increased. These clones were less permissive for HIV-1 than control clones transformed with the neo gene only. The antiviral state could be modulated by anti-IFN-beta antibodies, in that the continuous presence of antibodies in the culture medium abolished the enhanced resistance to HIV-1 replication, whereas the withdrawal of the antiserum restored the antiviral state, indicating that it did indeed result from the constitutive synthesis of human IFN-beta. These results demonstrate the possibility of restricting HIV-1 replication in human promonocytic cells by establishing SAVE. Further exploration of this method as a possible approach to somatic cell gene therapy of HIV infection appears worthwhile.     

Characteristics of behavior of knockout mice with genetic monoamine oxidase A deficiency

The effect of deletion of monoamine oxidase A (MAO A) in the gene encoding on behavior of transgenic Tg8 mice was studied. A decrease in the amplitude of acoustic startle reflex rather than the prepulse inhibition was found in lacking MAO A Tg8 mice, as compared with the control C3H strain. The exploratory activity in the hole-board test in Tg8 was decreased as well as the number of crossed lines in the light-dark test. Tg8 mice showed decreased latency and increased intensity of intermale aggression. At the same time, no difference was found between Tg8 and C3H mice in locomotor activity, in the expression of sexual motivation, and in the behavior in the elevated plus-maze test. No predisposition to catalepsy was shown.     

Spleen tyrosine kinase (Syk), a novel target of curcumin, is required for B lymphoma growth

Curcumin (diferuloylmethane), a component of dietary spice turmeric (Curcuma longa), has been shown in recent studies to have therapeutic potential in the treatment of cancer, diabetes, arthritis, and osteoporosis. We investigated the ability of curcumin to modulate the growth of B lymphomas. Curcumin inhibited the growth of both murine and human B lymphoma in vitro and murine B lymphoma in vivo. We also demonstrate that curcumin-mediated growth inhibition of B lymphoma is through inhibition of the survival kinase Akt and its key target Bad. However, in vitro kinase assays show that Akt is not a direct target of curcumin. We identified a novel target for curcumin in B lymphoma viz spleen tyrosine kinase (Syk). Syk is constitutively activated in primary tumors and B lymphoma cell lines and curcumin down-modulates Syk activity accompanied by down-regulation of Akt activation. Moreover, we show that overexpression of Akt, a target of Syk, or Bcl-x(L), a target of Akt can overcome curcumin-induced apoptosis of B lymphoma cells. These observations suggest a novel growth promoting role for Syk in lymphoma cells.