Cytochemistry and freeze-fracture of membranes isolated from the electrocyte of Electrophorus electricus (L.)

Summary Membranes were isolated from the main electric organ of Electrophorus electricus and studied by means of cytochemistry and freezefracture. The membrane fractions consisted of vesicles inside-in as determined by localization of anionic sites using colloidal iron and cationized ferritin particles. The anionic sites were not homogeneously distributed on the surface of the vesicle. Freeze-fracture showed the presence of intramembranous particles associated with either protoplasmic (P) or extracellular (E) faces of the membrane. Regions of the membrane without particles were observed. The results are discussed in relation to the existence of association between intramembranous particles and membrane receptors.For all correspondence     

Compound Autosomes in DROSOPHILA MELANOGASTER: The Meiotic Behavior of Compound Thirds

Studies of the meiotic distribution of compound-3 chromosomes in males and females of Drosophila melanogaster provided the following results. (1) From females homozygous for the standard arrangement of all chromosomes other than C(3L) and C(3R), less than 5% of the gametes recovered were nullosomic or disomic for compound-3 chromosomes. The frequency of nonsegregation differed between strains, but within a given strain it remained relatively constant. (2) According to egg-hatch frequencies, C(3L) and C(3R) segregate independently during spermatogenesis. (3) In females, structurally heterozygous second chromosomes occasion a marked increase in the recovery of nonsegregational progeny; in males, rearranged seconds have no apparent influence on the distribution of compound thirds. (4) The highest frequencies of nonsegregational progeny were recovered from C(3L);C(3R) females carrying compound-X (plus free Y) chromosomes. (5) In comparing the recovery of nonsegregating compound thirds to the recovery of rearranged heterologs, a definite nonrandom distribution was realized in several crosses. These results are examined in reference to the concepts of distributive pairing (Grell 1962). Moreover, considering the structural nature of compound autosomes, we propose that nonhomologous (distributive) pairing is a property of the centromeric region and suggest that rearrangements involving breaks in this region possibly alter the effectiveness of distributive pairing forces.     

An improved method for staining cells of the endocrine polypeptide (APUD) series by masked metachromasia: application of the principle of ''fixation by excluded volume''.

Synopsis Masked metachromasia is demonstrated by staining with a metachromatic basic dye, after acid hydrolysis of suitably fixed tissue. We report that the addition of 20% Carbowax 20M (an inert polymer, mol. wt. about 20000) to the hydrolysis mixture improved the reaction. The improved method gives increased metachromasia, greater tolerance to variations in hydrolysis conditions, and demonstrates a greater proportion of cells — presumably due to a lower threshold of sensitivity. Lower molecular weight polymers (Carbowax 1000, Carbowax 6000) are less effective.     

Prevalence of Edwards' syndrome. Clustering and seasonal variation?

Summary The incidence of Edwards' syndrome was found to be 1 per 4857 newborn children of 34000 consecutively newborn children in two Danish counties. Six of the 7 cases were born during the months of February through April.The incidence was high compared with the expected incidence of Edwards' syndrome of approximately 1 per 10000. This might be due to clustering in the area studied during the period 1967 to 1973.The finding of variations in incidence of children with Edwards' syndrome in different parts of the world, as well as the finding of seasonal variation in birth of such children, indicates that some of the etiological factors of nondisjunction of chromosome 18 are of an environmental nature.     

Cryptic phenology in plants: Case studies,implications, and recommendations

Plant phenology—the timing of cyclic or recurrent biological events in plants—offers insight into the ecology, evolution, and seasonality of plant‐mediated ecosystem processes. Traditionally studied phenologies are readily apparent, such as flowering events, germination timing, and season‐initiating budbreak. However, a broad range of phenologies that are fundamental to the ecology and evolution of plants, and to global biogeochemical cycles and climate change predictions, have been neglected because they are “cryptic”—that is, hidden from view (e.g., root production) or difficult to distinguish and interpret based on common measurements at typical scales of examination (e.g., leaf turnover in evergreen forests). We illustrate how capturing cryptic phenology can advance scientific understanding with two case studies: wood phenology in a deciduous forest of the northeastern USA and leaf phenology in tropical evergreen forests of Amazonia. Drawing on these case studies and other literature, we argue that conceptualizing and characterizing cryptic plant phenology is needed for understanding and accurate prediction at many scales from organisms to ecosystems. We recommend avenues of empirical and modeling research to accelerate discovery of cryptic phenological patterns, to understand their causes and consequences, and to represent these processes in terrestrial biosphere models.     

A high-throughput method for liquid chromatography–tandem mass spectrometry determination of plasma alkylresorcinols,biomarkers of whole grain wheat and rye intake

Plasma alkylresorcinols are increasingly analyzed in cohort studies to improve estimates of whole grain intake and their relationship with disease incidence. Current methods require large volumes of solvent (>10 ml/sample) and have relatively low daily sample throughput. We tested five different supported extraction methods for extracting alkylresorcinols from plasma and improved a normal-phase liquid chromatography coupled to a tandem mass spectrometer method to reduce sample analysis time. The method was validated and compared with gas chromatography–mass spectrometry analysis. Sample preparation with HybridSPE supported extraction was most effective for alkylresorcinol extraction, with recoveries of 77–82% from 100 μl of plasma. The use of 96-well plates allowed extraction of 160 samples per day. Using a 5-cm NH₂ column and heptane reduced run times to 3 min. The new method had a limit of detection and limit of quantification equivalent to 1.1–1.8 nmol/L and 3.5–6.1 nmol/L plasma, respectively, for the different alkylresorcinol homologues. Accuracy was 93–105%, and intra- and inter-batch precision values were 4–18% across different plasma concentrations. This method makes it possible to quantify plasma alkylresorcinols in 100 μl of plasma at a rate of at least 160 samples per day without the need for large volumes of organic solvents.     

Studying the uptake of cell-penetrating peptides

More than a decade ago, it was discovered that cationic peptides could traverse the cellular plasma membrane without specific transporter proteins or membrane damage. Subsequently, it was found that these peptides, known as cell-penetrating peptides (CPPs), were also capable of delivering cargos into cells, hence the great potential of these vectors was acknowledged. Today, many different research groups are working with CPPs, which necessitates efforts to develop unified assays enabling the comparison of data. Here we contribute three protocols for evaluation of CPPs which, if used in conjunction, provide complementary data about the amount and mechanism of uptake (fluorometric analysis and confocal microscopy, respectively), as well as the extent of degradation (HPLC analysis of cell lysates). All three protocols are based on the use of fluorescently labeled peptides and can be performed on the same workday.     

The return of an experimentally N-saturated boreal forest to an N-limited state: observations on the soil microbial community structure,biotic N retention capacity and gross N mineralisation

Background and aims

To find out how N-saturated forests can return to an N-limited state, we examined the recovery of biotic N sinks under decreasing N supply.

Methods

. We studied a 40-year-old experiment in Pinus sylvestris forest, with control plots, N0, three N treatments, N1-N3, of which N3 was stopped after 20 years, allowing observation of recovery.

Results

In N3, the N concentration in foliage was still slightly elevated, but the N uptake capacity of ectomycorrhizal (ECM) roots in N3 was no longer lower than in N0. Per area the amount of a biomarker for fungi, here mainly attributed ECM, was higher in N3 and N0 than in N1 and N2. Retention of labeled ¹⁵NH₄ ⁺ by the soil was greater in the control (99 %) and N3 (86 %), than in N1 (45 %) and N2 (29 %); we ascribe these differences to biotic retention because cation exchange capacity did not vary. Gross N mineralisation and retention of N correlated, negatively and positively, respectively, with abundance of ECM fungal biomarker.

Conclusions

. The results suggest a key role for ECM fungi in regulating the N cycle. We propose, in accordance with plant C allocation theory, that recovery is driven by increased tree below-ground C allocation to ECM roots and fungi.     

Economic impact of biofouling on a naval surface ship

In the present study, the overall economic impact of hull fouling on a mid-sized naval surface ship (Arleigh Burke-class destroyer DDG-51) has been analyzed. A range of costs associated with hull fouling was examined, including expenditures for fuel, hull coatings, hull coating application and removal, and hull cleaning. The results indicate that the primary cost associated with fouling is due to increased fuel consumption attributable to increased frictional drag. The costs related to hull cleaning and painting are much lower than the fuel costs. The overall cost associated with hull fouling for the Navy's present coating, cleaning, and fouling level is estimated to be $56M per year for the entire DDG-51 class or $1B over 15 years. The results of this study provide guidance as to the amount of money that can be reasonably spent for research, development, acquisition, and implementation of new technologies or management strategies to combat hull fouling.     

The protein-A plaque assay: a new system for the detection of cells secreting a given idiotype

With a modification of the protein-A plaque assay, cells secreting a given idiotype could be detected. Different anti-idiotypic antisera raised against the M-component of macroglobulinemia Waldenstr?m patients were used. The antibodies did not cross-react with cell clones of other patients or normal controls. Spontaneous plaques, without prior cultivation, were mainly shown to be of IgM class, the majority being idiotype specific, in blood and bone marrow lymphocytes from these patients. This is in contrast to the predominance of Ig or IgA found in normal blood donors. A low but recognizable stimulation of the malignant clone may be observed when cells were stimulated by certain polyclonal B cell activators.