Oviducal Contribution to Alteration of the Vitelline Coat in the Frog, Rana japonica. An Electron Microscopic Study

The vitelline coat (VC) surrounding coelomic eggs of the frog, Rana japonica , comprises bundles of filaments running both parallel and perpendicular to the egg surface. The coat gives little or no staining reaction with PA-CrA-Silver methenamine. In contrast, in the VC of uterine eggs the filament bundles are less conspicuous. and the interstices between the filament bundles stain strongly for carbohydrate. This alteration occurs during passage of the eggs down the first 1/20 th of the oviduct, the pars recta. The epithelium of the p. recta contains secretory cells, which contain electron-dense granules distinct from those in the jelly-secreting cells in more caudal portions of the oviduct. Treatment of coelomic eggs with an extract of p. recta followed by exposure to a sperm suspension resulted in marked swelling and softening of the VC. These results indicate that the contents of the granules secreted from the epithelial cells in the p. recta are deposited in the VC to increase its susceptibility to a fertilizing sperm.     

Studies on nitrosamine metabolism I. Subcellular distribution of radioactivity in tumor-susceptible tissues of RFM mice following administration of [14C] dimethylnitrosamine

The distribution of radioactivity in tumor-susceptible (liver and lung) and non-tumor-susceptible (heart, forestomach, and esophagus) tissues of male RFM mice was investigated at timed intervals following a single intragastric administration of ¹⁴C-labeled DMN.³ The greatest amount of radioactivity was associated with the tumor-susceptible tissues—liver and lung. At 15 min, the relative amount of radioactivity in the homogenates of heart, forestomach, esophagus, livers and lung was 1, 2, 3, 10, and 70, respectively. The AS components of lung contained about six times as much radioactivity as the liver 15 min after administration; at 16 hr, the level of radioactivity had decreased and was equal in amount. The AI components of both tumor-susceptible tissues incorporated much less radioactivity than the AS components, indicating that only a small amount of methyl label is covalently bound to cellular macromolecules. The amount or radioactivity in the AI components ranged from 2–34% in the lung and from 11–33% in the liver. In the lung C-fraction the range of radioactivity was 75–89% for the AS components and 52–74% for the AI components. The radioactivity in the AS components of liver C-fraction ranged from 50–89%, and from 52–68% for the AI components. The results suggest differences in the affinity, transport, and/or metabolism of DMN between liver and lung, as well as between tumor-susceptible and non-tumor-susceptible tissues.     

Cell Wall Solubilization in Pedicel Abscission of Begonia Flower Buds

Effects of metabolic inhibitors and growth regulators on the course of abscission and on the activities of cell wall solubilizing enzymes were studied in pedicel explants of Begonia flower buds. Actinomycin D, chloramphenicol and 2,4-dinitrophenol slightly retarded abscission, whereas cycloheximide exerted a strong inhibition if applied until 10.5 h after explant excision. Indoleacetic acid retarded and ethylene promoted abscission and cell wall solubilization. However, the activities of cell wall solubilizing enzymes did not correspond with the course of abscission. No polygalacturonase and pectic acid and pectin transeliminases could be detected in the abscission zone during abscission, whereas a low pectin methylesterase activity did not change. Endo- and exocellulase activities did not increase until about 10 h after the onset of abscission, indicating that they are the result rather than the cause of abscission.     

Hormonal Regulation of Pedicel Abscission in Begonia Flower Buds

In order to analyse the hormonal regulation of flower bud shedding in Begonia, levels of indoleacetic acid (IAA), abscisic acid (ABA) and ethylene were determined in buds and pedicels. The translocation and metabolism of ¹⁴C-labeled IAA in pedicel segments were also studied. In a monoecious Begonia fuchsioides hybrid, abscising male flower buds contain about 1% of the IAA present in non-abscising female flowers. In a male Begonia davisii hybrid, the seasonal variation in bud drop coincides with changes in the IAA content of the buds, while also the release of IAA from the bud to the pedicel is hampered. Abscission zones of these pedicels always contain abscission promoting ethylene concentrations. The tissue is prevented from responding with abscission by IAA from the flower buds. The buds also contain ABA but without influencing abscission considerably. Pretreatment with ethylene or ABA does not affect IAA transport in pedicel segments. The rate of this transport is 4–6 mm × h^–1:; the capacity increases with the transverse area. In young segments, IAA is decarboxylated and also otherwise metabolized.     

Evolutionary optimization of fluorescent proteins for intracellular FRET

Fluorescent proteins that exhibit Forster resonance energy transfer (FRET) have made a strong impact as they enable measurement of molecular-scale distances through changes in fluorescence. FRET-based approaches have enabled otherwise intractable measurements of molecular concentrations, binding interactions and catalytic activity, but are limited by the dynamic range and sensitivity of the donor-acceptor pair. To address this problem, we applied a quantitative evolutionary strategy using fluorescence-activated cell sorting to optimize a cyan-yellow fluorescent protein pair for FRET. The resulting pair, CyPet-YPet, exhibited a 20-fold ratiometric FRET signal change, as compared to threefold for the parental pair. The optimized FRET pair enabled high-throughput flow cytometric screening of cells undergoing caspase-3-dependent apoptosis. The CyPet-YPet energy transfer pair provides substantially improved sensitivity and dynamic range for a broad range of molecular imaging and screening applications.     

Factors associated with Diaphorina citri immigration into commercial citrus orchards in São Paulo State,Brazil

Seasonality often influences multiple aspects of vector-borne plant diseases. With respect to the management of plant disease vectors, an understanding of the factors driving seasonal changes in vector flight activity may improve management outcomes by facilitating more proactive application of control measures. One of the most challenging issues for citrus Huanglongbing management is to forecast when disease primary spread by bacteriliferous Diaphorina citri will occur. We monitored the temporal patterns of immigrating psyllids, using upwards of 1,200 traps spread among 9 citrus farms and checked on a weekly basis over 4 years. This dataset was analysed with a set of hierarchical models to estimate the effect of climatic variables on citrus foliage production (i.e. flush), and effects of climatic variables and citrus foliage dynamics on D. citri catches over the season. The results showed substantial seasonal variability in immigrating D. citri abundance, with the critical dispersal/migration period occurring between end of winter and spring. During this period, 65% of the total psyllids were collected on sticky traps. Seasonality of immigrating D. citri coincided with changes in certain climate variables, with negative effects of humidity and daily maximum temperatures, and a positive effect of rainfall amount in prior weeks. Maximum temperature and both daily minimum temperature and rainfall during prior weeks were also associated with new citrus flush production, which itself was positively related to immigrating D. citri abundance. Based on these results, citrus growers should be aware and intensify the frequency of psyllid control tactics (i.e. insecticide and/or kaolin sprays) during this period in order to prevent Ca. L. asiaticus transmission. These results are an important step towards developing the predictive framework needed to refine D. citri and huanglongbing management.     

PD123319 Augments Angiotensin II-Induced Abdominal Aortic Aneurysms through an AT2 Receptor-Independent Mechanism

Background

AT2 receptors have an unclear function on development of abdominal aortic aneurysms (AAAs), although a pharmacological approach using the AT2 receptor antagonist PD123319 has implicated a role. The purpose of the present study was to determine the role of AT2 receptors in AngII-induced AAAs using a combination of genetic and pharmacological approaches. We also defined effects of AT2 receptors in AngII-induced atherosclerosis and thoracic aortic aneurysms.

Methods and Results

Male AT2 receptor wild type (AT2 +/y) and deficient (AT2 -/y) mice in an LDL receptor −/− background were fed a saturated-fat enriched diet, and infused with either saline or AngII (500 ng/kg/min). AT2 receptor deficiency had no significant effect on systolic blood pressure during AngII-infusion. While AngII infusion induced AAAs, AT2 receptor deficiency did not significantly affect either maximal width of the suprarenal aorta or incidence of AAAs. The AT2 receptor antagonist PD123319 (3 mg/kg/day) and AngII were co-infused into male LDL receptor −/− mice that were either AT2 +/y or −/y. PD123319 had no significant effect on systolic blood pressure in either wild type or AT2 receptor deficient mice. Consistent with our previous findings, PD123319 increased AngII-induced AAAs. However, this effect of PD123319 occurred irrespective of AT2 receptor genotype. Neither AT2 receptor deficiency nor PD123319 had any significant effect on AngII-induced thoracic aortic aneurysms or atherosclerosis.

Conclusions

AT2 receptor deficiency does not affect AngII-induced AAAs, thoracic aortic aneurysms and atherosclerosis. PD123319 augments AngII-induced AAAs through an AT2 receptor-independent mechanism.     

A Smartphone Ecological Momentary Assessment/Intervention “App” for Collecting Real-Time Data and Promoting Self-Awareness

We have designed a flexible ecological momentary assessment/intervention smartphone (EMA/EMI) “app”. We examine the utility of this app for collecting real-time data, and assessing intra-subject variability, by using it to assess how freshman undergraduates spend their time. We also explore whether its use can promote greater self-awareness. Participants were randomly divided into an experimental group, who used the app, and a control group, who did not. We used the app to collect both randomized in-the-moment data as well as end-of-day data to assess time use. Using a posttest survey we asked participants questions about how they spent time throughout the school semester. We also asked the experimental group about their experience with the app. Among other findings, 80.49% participants indicated that they became more aware of how they spent their time using the app. Corroborating this report, among the experimental group, end-of-semester self-assessment of time spent wasted, and time spent using electronics recreationally, predicted semester GPA at a strength comparable to high school GPA and ACT score (two of the best single predictors for first semester college GPA), but had no correlation among controls. We discuss the advantages and limitations of using apps, such as ours, for EMA and/or EMI.     

Citrullus lanatus ‘sentinel’ (watermelon) extract reduces atherosclerosis in LDL receptor-deficient mice

Watermelon (Citrullus lanatus or C. lanatus) has many potentially bioactive compounds including citrulline, which may influence atherosclerosis. In this study, we determined the effects of C. lanatus, provided as an extract of the cultivar ‘sentinel,’ on hypercholesterolemia-induced atherosclerosis in mice. Male low-density lipoprotein receptor-deficient mice at 8 weeks old were given either C. lanatus ‘sentinel’ extract (2% vol/vol; n= 10) or a mixture of matching carbohydrates (2% vol/vol; n= 8) as the control in drinking water while being fed a saturated fat-enriched diet for 12 weeks ad libitum. Mice consuming C. lanatus ‘sentinel’ extract had significantly increased plasma citrulline concentrations. Systolic blood pressure was comparable between the two groups. Consumption of C. lanatus ‘sentinel’ extract led to lower body weight and fat mass without influencing lean mass. There were no differences in food and water intake and in urine output between the two groups. C. lanatus ‘sentinel’ extract administration decreased plasma cholesterol concentrations that were attributed to reductions of intermediate-/low-density lipoprotein cholesterol. Plasma concentrations of monocyte chemoattractant protein-1 and interferon-gamma were decreased and those of interleukin-10 were increased in mice consuming C. lanatus ‘sentinel’ extract. Intake of C. lanatus ‘sentinel’ extract resulted in reductions of atherosclerosis in both aortic arch and thoracic regions. In conclusion, consumption of C. lanatus ‘sentinel’ extract led to reduced body weight gain, decreased plasma cholesterol concentrations, improved homeostasis of pro- and anti-inflammatory cytokines, and attenuated development of atherosclerosis without affecting systolic blood pressure in hypercholesterolemic mice.