Plasma Testosterone and Testosterone Binding Affinities in Men with Impotence,Oligospermia, Azoospermia,and Hypogonadism

Mean plasma testosterone levels (± S.D.), using Sephadex LH-20 and competitive protein binding, were 629 ± 160 ng/100 ml for a group of 27 normal adult men, 650 ± 205 ng/100 ml for 27 impotent men with normal secondary sex characteristics, 644 ± 178 ng/100 ml for 20 men with oligospermia, and 563 ± 125 ng/100 ml for 16 azoospermic men. None of these values differ significantly. For 21 men with clinical evidence of hypogonadism the mean plasma testosterone (± S.D.), at 177 ± 122 ng/100 ml, differed significantly (P < 0·001) from that of the normal men.The mean testosterone binding affinities (as measured by the reciprocal of the quantity of plasma needed to bind 50% of ³H-testosterone tracer) were similar for normal, impotent, and oligospermic men. Though lower for azoospermic men the difference was not significant (P >0·1). For 12 of the 16 hypogonadal males the testosterone binding affinity was normal, but raised binding affinities, similar to those found in normal adult females or prepubertal boys (about twice normal adult male levels), were found in four cases of delayed puberty. These findings help to explain why androgen therapy is usually useless in the treatment of impotence.     

Secretory Activity Is Rapidly Induced in Stigmatic Papillae by Compatible Pollen,but Inhibited for Self-Incompatible Pollen in the Brassicaceae

[In the Brassicaceae, targeted exocytosis to the stigmatic papillar plasma membrane under the compatible pollen grain is hypothesized to be essential for pollen hydration and pollen tube penetration. In contrast, polarized secretion is proposed to be inhibited in the stigmatic papillae during the rejection of self-incompatible pollen. Using transmission electron microscopy (TEM), we performed a detailed time-course of post-pollination events to view the cytological responses of the stigmatic papillae to compatible and self-incompatible pollinations. For compatible pollinations in Arabidopsis thaliana and Arabidopsis lyrata, vesicle secretion was observed at the stigmatic papillar plasma membrane under the pollen grain while Brassica napus stigmatic papillae appeared to use multivesicular bodies (MVBs) for secretion. Exo70A1, a component of the exocyst complex, has been previously implicated in the compatible pollen responses, and disruption of Exo70A1 in both A. thaliana and B. napus resulted in a loss of secretory vesicles/MVBs at the stigmatic papillar plasma membrane. Similarly, for self-incompatible pollinations, secretory vesicles/MVBs were absent from the stigmatic papillar plasma membrane in A. lyrata and B. napus; and furthermore, autophagy appeared to be induced to direct vesicles/MVBs to the vacuole for degradation. Thus, these findings support a model where the basal pollen recognition pathway in the stigmatic papilla promotes exocytosis to accept compatible pollen, and the basal pollen recognition pathway is overridden by the self-incompatibility pathway to prevent exocytosis and reject self-pollen.     

Effects of Burning and Grazing on a Coastal California Grassland

We tested the effects of fall burning and protection from livestock grazing as management to enhance native grasses on a coastal grassland in central California. Plants from the Mediterranean, introduced beginning in the late 1700s, have invaded and now dominate most of California's grasslands. Coastal grasslands are generally less degraded than those inland and have higher potential for restoration and conservation. Productivity of the experimental plots varied annually and declined over the course of the study because of rainfall patterns. Foliar cover of the native Danthonia californica (California oatgrass) increased more under grazing than grazing exclusion and did not respond to burning. Two other natives, Nassella pulchra (purple needlegrass) and Nassella lepida (foothill needlegrass), responded variably to treatments. The response of N. pulchra differed from that reported on more inland sites in California. Restoring these grasslands is complicated by differing responses of target species to protection from grazing and burning. The current practice of managing to enhance single species of native plants (e.g., N. pulchra) may be detrimental to other equally important native species.     

Secondary (iso)BAs cooperate with endogenous ligands to activate FXR under physiological and pathological conditions

IsoBAs, stereoisomers of primary and secondary BAs, are found in feces and plasma of human individuals. BA signaling via the nuclear receptor FXR is crucial for regulation of hepatic and intestinal physiology/pathophysiology. Aim: Investigate the ability of BA-stereoisomers to bind and modulate FXR under physiological/pathological conditions. Methods: Expression-profiling, luciferase-assays, fluorescence-based coactivator-association assays, administration of (iso)-BAs to WT and cholestatic mice. Results: Compared to CDCA/isoCDCA, administration of DCA/isoDCA, UDCA/isoUDCA only slightly increased mRNA expression of FXR target genes; the induction was more evident looking at pre-mRNAs. Notably, almost 50% of isoBAs were metabolized to 3-oxo-BAs within 4 h in cell-based assays, making it difficult to study their actions. FRET-based real-time monitoring of FXR activity revealed that isoCDCA>CDCA stimulated FXR, and isoDCA and isoUDCA allowed fully activated FXR to be re-stimulated by a second dose of GW4064. In vivo co-administration of a single dose of isoBAs followed by GW4064 cooperatively activated FXR, as did feeding of UDCA in a background of endogenous FXR ligands. However, in animals with biliary obstruction and concomitant loss of intestinal BAs, UDCA was unable to increase intestinal Fgf15. In contrast, mice with an impaired enterohepatic circulation of BAs (Asbt?/?, Ostα?/?), administration of UDCA was still able to induce ileal Fgf15 and repress hepatic BA-synthesis, arguing that UDCA is only effective in the presence of endogenous FXR ligands. Conclusion: Secondary (iso)BAs cooperatively activate FXR in the presence of endogenous BAs, which is important to consider in diseases linked to disturbances in BA enterohepatic cycling.     

Generation of an immortal differentiated lung type-II epithelial cell line from the adult H-2KbtsA58 transgenic mouse

Summary This paper describes a new fully differentiated Type-II alveolar epithelial cell line designated T₇, derived from transgenic H-2K^b-tsA58 mice, capable of being passaged as an immortalized cloned cell line in culture. H-2K^b-tsA58 mice harbor a temperature-sensitive (ts) mutant of the simian virus 40 (SV40) large tumor antigen (T antigen) under the control of the γ-interferon (INF)-inducible mouse major histocompatibility complex H-2K^b promoter. When cultured under permissive conditions (33°C and in the presence of γ-INF) cells isolated from H-2K^b-tsA58 mice express the large T antigen, which drives the cells to proliferate. However, upon withdrawal of the γ-INF and transfer of the cells to a higher temperature (39°C), T antigen expression is turned off, the cells stop proliferating and differentiate. The T₇ cell line is a clonal cell line originally derived from a Type-II cell-rich fraction isolated from lungs of H-2K^b-tsA58 mice. The T₇ cells form confluent monolayers, and have a polarized epithelial cell morphology with tight junctions and apical microvilli. In addition, the T₇ cells have distinct cytoplasmic lamellar bodies, which become more numerous and pronounced when the cells are grown under nonpermissive conditions. The T₇ cells synthesize and secrete phosphatidylcholine and the three surfactant proteins, SP-A, SP-B, and SP-C. The T₇ cell line is unique in that it is the first non-tumor-derived Type-II cell line capable of synthesizing and secreting the major components of surfactant. Based on the criteria studied, the T₇ cell line is phenotypically very similar to normal Type-II cells. The T₇ cell line, therefore, should prove a valuable experimental system to advance the study of the cell biology/physiology of surfactant metabolism and secretion as well as serve as a model for other studies of Type-II cell physiology.     

Detection of Biosignatures by Geomatrix-Assisted Laser Desorption/Ionization (GALDI) Mass Spectrometry

Identification of mineral-associated biosignatures is of significance for retrieving biochemical information from geological records here on Earth and for detecting signs of life on other planets, such as Mars. An investigation using laser desorption Fourier transform mass spectrometry was conducted to determine whether geomatrix-assisted laser desorption/ionization (GALDI) can be used to detect amino acids (e.g., histidine, threonine, and cysteine) and small proteins (e.g., gramicidin S) associated with mineral phases and whether the geomatrix impacts detection. Iron oxide (Fe₂ O ₃ ) and sodium chloride (NaCl) were investigated as clean chemical analogues of hematite and halite, respectively, which have both been detected on the surface of Mars. Samples were prepared by 2 methods: (1) application of analyte solution to the geomatrix surface with subsequent drying; and (2) physical mixing of analyte and geomatrix. Amino acids incorporated within NaCl by physical mixing yielded a better signal-to-noise ratio than those that were applied to the surface of a NaCl pellet. The composition of the geomatrix had an influence on the detection of biomolecules. Peaks corresponding to the cation-attached biomolecular ions were observed for the NaCl prepared samples. However, no biomolecular ion species were observed in samples using Fe ₂ O ₃ as geomatrix. Instead, only minor peaks that may correspond to ions derived from fragments of the biomolecules were obtained.     

Extended models of the ventilatory response to sustained isocapnic hypoxia in humans

Liang, Pei-Ji, Daphne A. Bascom, and Peter A. Robbins.Extended models of the ventilatory response to sustained isocapnic hypoxia in humans. J. Appl. Physiol. 82(2): 667-677, 1997.The purpose of this study was to examine extensions of a modelof hypoxic ventilatory decline (HVD) in humans. In the original model (model I) devised by R. Painter, S. Khamnei, and P. Robbins(J. Appl. Physiol. 74: 2007-2015, 1993), HVD is modeledentirely by a modulation of peripheral chemoreflex sensitivity. In thefirst extension (model II), a more complicated dynamic is usedfor the change in peripheral chemoreflex sensitivity. In the secondextension (model III), HVD is modeled as a combination ofboth the mechanism of Painter et al. and a component that isindependent of peripheral chemoreflex sensitivity. In all cases, aparallel noise structure was incorporated to describe the stochasticproperties of the ventilatory behavior to remove the correlation of theresiduals. Data came from six subjects from a study by D. A. Bascom, J. J. Pandit, I. D. Clement, and P. A. Robbins (Respir. Physiol.88: 299-312, 1992). For model II, there was a significantimprovement in fit for two out of six subjects. The reasons for thiswere not entirely clear. For model III, the fit was againsignificantly improved in two subjects, but in this case the subjectswere those who had the most marked undershoot and recovery ofventilation at the relief of hypoxia. In these two subjects, thechemoreflex-independent component contributed ~50% to total HVD.In the other four subjects, the chemoreflex-independent componentcontributed ~10% to total HVD. It is concluded that in somesubjects, but not in others, there may be a component of HVD thatis independent of peripheral chemoreflex sensitivity.

     

IPS-1 Is Essential for the Control of West Nile Virus Infection and Immunity

The innate immune response is essential for controlling West Nile virus (WNV) infection but how this response is propagated and regulates adaptive immunity in vivo are not defined. Herein, we show that IPS-1, the central adaptor protein to RIG-I-like receptor (RLR) signaling, is essential for triggering of innate immunity and for effective development and regulation of adaptive immunity against pathogenic WNV. IPS-1^−/− mice exhibited increased susceptibility to WNV infection marked by enhanced viral replication and dissemination with early viral entry into the CNS. Infection of cultured bone-marrow (BM) derived dendritic cells (DCs), macrophages (Macs), and primary cortical neurons showed that the IPS-1-dependent RLR signaling was essential for triggering IFN defenses and controlling virus replication in these key target cells of infection. Intriguingly, infected IPS-1^−/− mice displayed uncontrolled inflammation that included elevated systemic type I IFN, proinflammatory cytokine and chemokine responses, increased numbers of inflammatory DCs, enhanced humoral responses marked by complete loss of virus neutralization activity, and increased numbers of virus-specific CD8+ T cells and non-specific immune cell proliferation in the periphery and in the CNS. This uncontrolled inflammatory response was associated with a lack of regulatory T cell expansion that normally occurs during acute WNV infection. Thus, the enhanced inflammatory response in the absence of IPS-1 was coupled with a failure to protect against WNV infection. Our data define an innate/adaptive immune interface mediated through IPS-1-dependent RLR signaling that regulates the quantity, quality, and balance of the immune response to WNV infection.     

Latitudinal cline of requirement for far‐red light for the photoperiodic control of budset and extension growth in Picea abies (Norway spruce)

To test for the effects of far‐red light on preventing budset in Picea abies , seedlings of six populations originating from latitudes between 67°N and 47°N were grown for 4–8 weeks in continuous incandescent (metal halogen) light at 300 µmol m⁻² s⁻¹ and 20°C and then transferred, at the same temperature, to a daily regime of 8 h incandescent light (300 µmol m⁻² s⁻¹) followed by 16 h cool white fluorescent light (40 µmol m⁻² s⁻¹). (Cool white lamps are deficient in far‐red light, with a R/FR ratio of 7.5 compared with 2.0 for the incandescent lamps.) All the seedlings from 67° and 80% of those from 64° stopped extension growth and set terminal buds within 28 days of the change of regime. The seedlings from 61° and further south continued growing, as did control seedlings from 67° grown as above but with incandescent light at 20 µmol m⁻² s⁻¹ replacing cool white illumination. To distinguish between a clinal and ecotypic pattern of variation, the interval between 64° and 59° was investigated by growing populations originating from that area in the same regimes as before. After 28 days in the cool white day‐extension regime, the percentage budset was 86 for the population from 64°, 0 for the population from 59° and 25–50 for the intermediate populations; i.e. the populations showed a clinal variation in requirement for far‐red light according to latitude. Thus northern populations of Picea abies appear to behave as 'light‐dominant' plants for the photoperiodic control of extension growth and budset, whereas the more southern populations behave as 'dark‐dominant' plants.