Nuclear factor-kappa B localization and function within intrauterine tissues from term and preterm labor and cultured fetal membranes

Background  

The objective of this study was to quantify the nuclear localization and DNA binding activity of p65, the major transactivating nuclear factor-kappa B (NF-kappaB) subunit, in full-thickness fetal membranes (FM) and myometrium in the absence or presence of term or preterm labor.     

Towards many colors in FISH on 3D-preserved interphase nuclei

The article reviews the existing methods of multicolor FISH on nuclear targets, first of all, interphase chromosomes. FISH proper and image acquisition are considered as two related components of a single process. We discuss (1) M-FISH (combinatorial labeling + deconvolution + wide-field microscopy); (2) multicolor labeling + SIM (structured illumination microscopy); (3) the standard approach to multicolor FISH + CLSM (confocal laser scanning microscopy; one fluorochrome - one color channel); (4) combinatorial labeling + CLSM; (5) non-combinatorial labeling + CLSM + linear unmixing. Two related issues, deconvolution of images acquired with CLSM and correction of data for chromatic Z-shift, are also discussed. All methods are illustrated with practical examples. Finally, several rules of thumb helping to choose an optimal labeling + microscopy combination for the planned experiment are suggested.     

A new method for quantifying encephalization in the growing individual

Here we describe a new method for quantifying encephalization in the growing individual and provide a worked example of the methods. The new method is based on the use of conditional SD scores derived from brain and body growth references. These encephalization SD scores control for age, sex and body size effects on brain size, and therefore, control for the confounds associated with allometry as well as growth differences between the brain and body and between the sexes. The methods also control for distribution skewness. Encephalization SD scores derived from pre- and post-natal data may be directly compared and changes in SD score over time assessed. These methods may be applied to a broad range of data where relative size during growth is to be quantified. Derived SD scores may also be applied to correlation and regression analyses where statistical relationships with other variables are of interest.     

A case-control follow-up study for disease-specific mortality

Case-control studies often rely on subjects to report their own screening or exposure information: this information is often obtained from cases after the event of interest has occurred. This is problematic for mortality outcomes, because dead subjects cannot report the desired information. To avoid this problem, Weiss and Lazovich (1996, American Journal of Epidemiology 143, 319-322) proposed obtaining exposure or screening information from potential cases, i.e., subjects diagnosed with disease, at the time of disease diagnosis, and also from a referent series. The design is best viewed as a new scheme for sampling from a cohort. I review estimation of the effects of time-varying screening or exposure in cohort studies, using a new factorization. I then show how this factorization, together with ignorability assumptions, allows valid estimation from these new designs. Even when the sampling fraction of nondiseased subjects is unknown, causal risk ratios are estimable if diagnosis is rare in the cohort. I illustrate and compare conventional and new methods with data from the Health Insurance Plan study.     

Ambulatory estimates of maximal aerobic power from foot -ground contact times and heart rates in running humans.

Seeking to develop a simple ambulatory test of maximal aerobic power (VO(2 max)), we hypothesized that the ratio of inverse foot-ground contact time (1/t(c)) to heart rate (HR) during steady-speed running would accurately predict VO(2 max). Given the direct relationship between 1/t(c) and mass-specific O(2) uptake during running, the ratio 1/t(c). HR should reflect mass-specific O(2) pulse and, in turn, aerobic power. We divided 36 volunteers into matched experimental and validation groups. VO(2 max) was determined by a treadmill test to volitional fatigue. Ambulatory monitors on the shoe and chest recorded foot-ground contact time (t(c)) and steady-state HR, respectively, at a series of submaximal running speeds. In the experimental group, aerobic fitness index (1/t(c). HR) was nearly constant across running speed and correlated with VO(2 max) (r = 0.90). The regression equation derived from data from the experimental group predicted VO(2 max) from the 1/t(c). HR values in the validation group within 8.3% and 4.7 ml O(2) x kg(-1) x min(-1) (r = 0.84) of measured values. We conclude that simultaneous measurements of foot-ground constant times and heart rates during level running at a freely chosen constant speed can provide accurate estimates of maximal aerobic power.     

Unordered arrangement of chromosomes in the nuclei of chicken spermatozoa

The arrangement of chromosomes in the elongated sperm nuclei of chicken was studied using fluorescence in situ hybridization with probes specific for telomeres of all chromosomes, a microchromosome, the long arm of chromosome 6, the large heterochromatic block on the Z-chromosome, and the same heterochromatic block plus subtelomeric sites on macrochromosomes 1–4. The positions of all probes vary from one sperm to another. No order in chromosome arrangement is apparent. It is suggested that large chromosome size and small chromosome number correlate with constant positions of chromosomes and vice versa. Based on the known quantity of repetitive units of the repeat on the Z-chromosome, the degree of compaction of chromatin in the chicken sperm nucleus is estimated as ca 0.7 Mb/μm. As judged from the length of the heterochromatic region of the Z-chromosome at the lampbrush stage, the total length of the Z-chromosome in mature sperm is 2.5–4 times that of the sperm nucleus. Received: 15 December 1997; in revised form: 24 March 1998 / Accepted: 14 April 1998     

The GAIF-Positive Population of Neurons in the Evolution of the Temnocephalida

The nervous system of three species of the Temnocephalida has been studied using the GAIF method (which in small flatworms mostly reveals sensory catecholaminergic neurons). These species represent the evolution from the primitive “turbellarian-like” temnocephalids to the most specialised ones with tentacles and a sucker. The numbers and positions of GAIF-positive neurons are invariant within each species and do not change from hatching to full maturity. A characteristic unpaired neuron contributing to the innervation of the anterior margin of the body is present in all species: such a cell has previously been found only in marine Thalassovortex tyrrhenicus (Dalyellidae) which confirms close relationships between these taxa. Our series of species shows (i) a reduction in number of GAIF-positive perikarya associated with the lateral cords and reduction of GAIF-positive innervation on the ventral side of the body, which is probably related to the loss of ciliary locomotion (the shift to passive hunting and looping locomotion) and (ii) reinforcement of the GAIF-positive innervation of the anterior end of the body which begins to play an important role in capturing the prey. The retention of the medial unpaired neuron and nearly identical sets of GAIF-positive neurons in Diceratocephala boschmai and Craspedella pedum (rather different in morphology) give the first indication (in the Plathelminthes) of persistence of homologous neurons through significant evolutionary transformations of the organs they innervate.     

Sensory cells in the sucker of Craspedella pedum (Plathelminthes, Temnocephalida): in vivo staining with DiO and SEM and TEM observations

 Two pairs of identified sensory neurons innervating the sucker of Craspedella pedum (designated ADS1, ADS2) were found to accumulate DiO (3,3′-dioctadecyloxacarbocyanine perchlorate) in vivo. The number, position and morphology of these neurons do not change throughout the postembryonic period of life. The axons of the ADS cells run forward within the ventral cords and their dendrites are parallel. They enter the sucker, ramify and terminate in numerous sensory endings in a wide peripheral zone of the disc. SEM reveals a single type of sensilla: small spot-like structures with several short cilia. They are scattered within the zone accommodating the openings of the adhesive glands and their distribution corresponds to that of the stained terminals. TEM observations (including about 20 full reconstructions from serial ultrathin sections) show five types of sensory endings on the disc with the following structures: (1) a short cilium and thin rootlet, (2) aciliary with a normal rootlet and a club-shaped apical portion, (3) aciliary with a club-shaped apical portion and a body similar to the apical part of the rootlet, (4) aciliary with large apical granule and (5) aciliary with small apical granules. Type 2–5 receptors form a morphological series suggesting that they are stages of formation of the common type 4 receptor. Not fully formed type 1 receptors have been found within the epidermis in adult animals. This suggests that, although ADS perikarya persist throughout the life of the animal, the nerve endings they form might be constantly renewed. Judging from the morphological and behavioural data, the functions of the ADS neurons might include: (1) monitoring of the close contact between the surface of the sucker and the substratum prior to adhesion and (2) checking the viscosity of the adhesive secretion prior to release of the sucker. Accepted: 16 December 1997