Response of late-instar Blattella germanica (Dictyoptera: Blattellidae) to dietary insect growth regulators

Late-instar German cockroaches, Blattella germanica (L.), were used to evaluate the relative effects of single treatments and combinations of three insect growth regulators (IGRs): pyriproxyfen, fenoxycarb, and diflubenzuron. Groups of 15 males or 15 females were held for 2 wk on food treated with varying amounts of IGRs. After removal, newly mature adults were placed with untreated adults of the opposite sex. Mortality, adult phenotype (normal wings, divergent wings, curly wings, and nymphoids), and reproduction were assessed. Mortality occurred largely in the nymphal stage except when all three IGRs were combined. Neither the average number of nymphs per egg case nor hatch of egg cases from phenotypically normal cockroaches (normal wings, occasional darkening of the body) was affected in single treatments or in combinations of two IGRs, but nymphal numbers were reduced when the three IGRs were combined. Hatch from matings of insects with divergent wings varied. At low concentrations (3-10 ppm), hatch was generally normal; at high concentrations, most mating tests were unproductive. Mating tests of cockroaches with curly wings were almost always unproductive. When males with curly wings were mated, females dropped either unfertilized egg cases (no mating) or partially fertilized egg cases. Curly-wing females either dropped unfertilized egg cases or failed to form egg cases because of deleterious effects on ovarian development. Nymphoids did not mate. Diflubenzuron at 100 ppm had no effect other than causing the appearance of a few insects with divergent wings. Effects on phenotype and reproduction began at 3 ppm of both pyriproxyfen and fenoxycarb and at a comparable concentration in the combination of pyriproxyfen + fenoxycarb (1 ppm each). Female sterility was complete at 100 ppm of pyriproxyfen and fenoxycarb. When pyriproxyfen or fenoxycarb was combined with equal amounts of diflubenzuron, the number of productive matings was not reduced at 6 ppm (3 ppm per each IGR). At 20 ppm (10 ppm per each IGR), a reduction in productive matings coincided with the appearance of curly wings. Complete female sterility occurred only at 600 ppm (300 ppm per IGR). The most severe effects occurred in the experiment with equal amounts of pyriproxyfen, fenoxycarb, and diflubenzuron. In addition to reduced hatch from normal phenotypes, this experiment caused complete male sterility (300 ppm; 100 ppm of each IGR). Female sterility was complete at greater than or equal to 90 ppm (30 ppm of each IGR).     

Prefreezing and post-thaw semen characteristics of five ram breeds collected by electroejaculation

Rams representing five breeds were electroejaculated twice weekly, during a three-week collection period. Ejaculates were evaluated for volume and concentration before freezing and for rate of motility and percentages of motile and abnormal cells both before and after freezing. Interactions between breed and collection period were evident (P<0.05) for semen volume and post-thaw values for rate of motility and percentage motile cells. Breeds differed (P<0.05) in these traits during some periods. In contrast, pre-freezing observations of rate of motility, percentage motile and abnormal cells and post-thaw percentage abnormal cells did not differ (P>0.15) among breeds. Sperm concentration per ejaculate tended to vary (P=0.11) among breeds. Semen characteristics frequently varied across collection periods. Rams within a breed differed (P<0.01) in all semen traits except post-thaw rate of motility and percentage motile cells. Semen was negatively affected by the freezing and thawing procedure. Ram within a breed and ejaculate within ram should be considered when selecting electroejaculated semen for freezing and subsequent use in artificial insemination.     

The E3 ubiquitin ligase,HECTD1, is involved in ABCA1-mediated cholesterol export from macrophages

The ABC lipid transporters, ABCA1 and ABCG1, are essential for maintaining lipid homeostasis in cells such as macrophages by exporting excess cholesterol to extracellular acceptors. These transporters are highly regulated at the post-translational level, including protein ubiquitination. Our aim was to investigate the role of the E3 ubiquitin ligase HECTD1, recently identified as associated with ABCG1, on ABCG1 and ABCA1 protein levels and cholesterol export function. Here, we show that HECTD1 protein is widely expressed in a range of human and murine primary cells and cell lines, including macrophages, neuronal cells and insulin secreting β-cells. siRNA knockdown of HECTD1 unexpectedly decreased overexpressed ABCG1 protein levels and cell growth, but increased native ABCA1 protein in CHO-K1 cells. Knockdown of HECTD1 in unloaded THP-1 macrophages did not affect ABCG1 but significantly increased ABCA1 protein levels, in wild-type as well as THP-1 cells that do not express ABCG1. Cholesterol export from macrophages to apoA-I over time was increased after knockdown of HECTD1, however these effects were not sustained in cholesterol-loaded cells. In conclusion, we have identified a new candidate, the E3 ubiquitin ligase HECTD1, that may be involved in the regulation of ABCA1-mediated cholesterol export from unloaded macrophages to apoA-I. The exact mechanism by which this ligase affects this pathway remains to be elucidated.     

Receptor binding of PDGF-AA and PDGF-BB,and the modulation of PDGF receptors by TGF-β, in human periodontal ligament cells

The growth factors PDGF-AA and PDGF-BB have previously been shown to be potent mitogens for human periodontal ligament (hPDL) cells in vitro. Additionally, the mitogenic response to PDGF-AA has been shown to be specifically inhibited by TGF-β. The purpose of the present investigation was to examine the binding of PDGF-AA and PDGF-BB, and the modulation of PDGF binding by TGF-β, in hPDL cells. Scatchard analysis identified an average of 32,000 PDGF-AA high-affinity binding sites per cell with a dissociation constant (K_d) of 0.66 nM and an average of 36,000 PDGF-BB binding sites per cell with a dissociation constant (k_d) of 0.44 nM. After treatment with TGF-β, the receptor number for PDGF-AA was found to specifically decrease by approximately 50%, with no change in binding affinity. This reduced number of binding sites was shown to correlate with both a decrease in levels of receptor tyrosine phosphorylation and a decreased number of α receptor subunits. Northern blot analysis identified the TGF-β-mediated decrease in PDGF α receptor subunit mRNA levels. PDGF-BB showed little change in the number of binding sites or in the binding affinity with TGF-β treatment, and the data were consistent with an increase in the number of β receptor subunits. These results demonstrate nearly equivalent numbers of receptors for both PDGF-AA and PDGF-BB in hPDL cells. Also, modulation of PDGF binding, by TFG-β, was shown to result in a reduced number of α receptor subunits with an increase in the number of β receptor subunits. © 1995 Wiley-Liss, Inc.     

Ultrastructural localization of the high molecular weight proteins associated with in vitro-assembled brain microtubules

Microtubules isolated from brain extracts by in vitro assembly (1, 19, 23) are composed principally of two tubulins and two high molecular weight proteins (microtubule-associated proteins [MAPS] 1 and 2) (2,5,7,20). Recently, it was demonstrated that in vitro-assembled brain microtubules (neurotubules) are coated with filaments (5, 7) which are similar to the filaments attached to neurotubules in situ (4, 15, 21, 24, 25), and it was suggested that the filaments are composed of the higher molecular weight MAPs (5, 7, 12). In this study, microtubules were assembled in the presence and absence of the MAPs, and thin sections of the microtubules were examined by electron microscopy. The results show that the filaments only occur on microtubules assembled in the presence of the MAPs and it is therefore concluded that the filaments are composed of the high molecular weight MAP's.     

Stage-specific embryonic antigens (SSEA-3 and -4) are epitopes of a unique globo-series ganglioside isolated from human teratocarcinoma cells.

Two monoclonal antibodies (MC631 and MC813-70) raised against 4- to 8-cell stage mouse embryos and a human teratocarcinoma cell line, respectively, detect the stage-specific embryonic antigens, the previously defined SSEA-3 and SSEA-4, described herein. These antibodies were both reactive with a unique globo-series ganglioside with the structure shown below: (formula; see text) The antibodies were found to recognize sequential regions of this ganglioside, i.e., MC813-70 recognizes the terminal 'a' structure whereas antibody MC631 recognizes the internal 'b' structure. Thus, a set of two antibodies defines this unique embryonic antigen. During differentiation of human teratocarcinoma 2102Ep cells, the globo-series glycolipids defined by these antibodies decrease and the lacto-series glycolipids, reacting with the SSEA-1 antibody appear. This antigenic conversion suggests that a shift of glycolipid synthesis from globo-series to lacto-series glycolipids occurs during differentiation of human teratocarcinoma and perhaps of pre-implantation mouse embryos.     

Experimental Scrapie in the Mouse: Electrophoretic and Sedimentation Properties of the Partially Purified Agent

Abstract: High resolution purification procedures for the separation and characterization of neuropeptides are described. The methods, which are complementary, are based on ion exchange chromatography and reverse phase high pressure liquid chromatography and are designed to be used in conjunction with radioimmunoassays and bioassays for known peptides or structural identification in the case of peptides of unknown sequence. The high pressure liquid chromatographic method is versatile, highly reproducible and applicable to picomolar quantities of peptides and it gives very high resolution between most of the neuropeptides of present-day interest. The applicability of the method to the study of crude brain extracts is demonstrated.     

Dimethyl sulfoxide: reversible inhibitor of pollen tube growth

Five percent dimethyl sulfoxide (DMSO) completely inhibited tube initiation, stopped tube growth and suppressed the high respiration associated with tube growth of lily pollen. The effect of DMSO on respiration was indirect because uncoupling concentrations of 2,4-dinitrophenol abolished the inhibition of respiration. Five percent DMSO did not inhibit rapid starch synthesis during the first 30 minutes of incubation, nor did DMSO inhibit the period of high respiration associated with rapid starch synthesis. DMSO did not cause permanent damage to the cells since normal pollen tube growth occurred after its removal. Dimethyl sulfoxide is not a general inhibitor of pollen metabolism, but it may be a specific inhibitor of a process required for tube growth.     

An Infrared Spectrophotometric Study of Some Strains of Staphylococcus aureus

A method of obtaining expanded differential spectra of staphylococcal strains for comparative studies is described. Some differences found through the comparison of a large number of strains isolated from humans, turkeys, and other hosts are shown. No spectral characteristics that would associate a particular strain with its host were discerned. Differences in the strength of a band at 11.6 to 11.7 μ allowed division of the strains examined into two groups.