Micropropagation of Cowania subintegra and Cowania stansburiana

Both Cowania subintegra Kearney and C. stansburiana Torr. were successfully propagated in vitro. Shoot proliferation occurred from shoot tips of green-house grown C. subintegra using a modified Murashige and Skoog medium supplemented with 4.4 M 6-benzyladenine and 0.5 M indole butyric acid. Excised microshoots (1.5–3.0 cm long) of both species were rooted using a two-step process in which they were cultured for 3 days in a root initiation medium with 2.7 M naphthaleneacetic acid and then transferred to a low nitrogen root elongation medium without auxin. Plantlets were successfully transferred to soilless potting mix.     

BRCA1 maps proximal to D17S579 on chromosome 17q21 by genetic analysis

Previous studies have demonstrated linkage between early-onset breast cancer and ovarian cancer and genetic markers on chromosome 17q21. These markers define the location of a gene (BRCA1) which appears to be inherited as an autosomal dominant susceptibility allele. We analyzed five families with multiple affected individuals for evidence of linkage to the BRCA1 region. Two of the five families appear to be linked to BRCA1. One apparently linked family contains critical recombinants, suggesting that the gene is proximal to the marker D17S579 (Mfd188). These findings are consistent with the maximum-likelihood position estimated by the Breast Cancer Linkage Consortium and with recombination events detected in other linked families. Linkage analysis was greatly aided by PCR-based analysis of paraffin-embedded normal breast tissue from deceased family members, demonstrating the feasibility and importance of this approach. One of the two families with evidence of linkage between breast cancer and genetic markers flanking BRCA1 represents the first such family of African-American descent to be reported in detail.     

Topography of short portal vessels in the rat pituitary gland: a scanning electron-microscopic and morphometric study of corrosion cast replicas

We applied scanning electron microscopy combined with imaging and morphometric techniques to analyze the dorsal topography and morphology of short portal vessels linking the capillary beds of the pituitary neural and anterior lobes in adult male albino rats. The pituitary microvasculature was replicated by intracarotid injection of Batson's No. 17 compound producing plastic casts that were advantageous for comprehensive morphometric analyses using an imaging device. The analysis revealed the existence of two types of portal vessels having quantitatively different morphological properties. The bilateral venular plexus of 3–4 vessels located at the base of the infundibular stalk (each venule measuring 300 m in length and 32 m in diameter) appears to be the major part of the short portal system in the dorsum of the rat pituitary gland. Narrower capillary-like shunt vessels (6.8 m in diameter), of about the same length as the venules, were situated throughout other subregions of the intermediate lobe cleft. The short portal vessels of both types made direct anastomoses with the capillary networks in the neural and anterior lobes. The neural lobe capillaries were twice as numerous (1324 per mm²), and only half as wide (6.2 m), as the sinusoidal capillaries in the anterior lobe (density of 637 per mm²; diameter of 13.7 m). The topographical position of the portal venular system suggests that the caudolateral subregions of the pituitary neural and anterior lobes have a functional relationship dependent on rapid interlobe transfer of neurohumoral factors such as hormones via the portal blood. This process appears to be supplemented throughout the rest of the cleft between the two lobes by a small number of capillary shunts that supply the epithelial cell lobules of the intermediate lobe in situ. The findings collectively indicate that this portal system provides a constant stream of neurohumoral information that is shared moment-by-moment between the pituitary neural and anterior lobes.     

Latent group detection in functional partially linear regression models

In this paper, we propose a functional partially linear regression model with latent group structures to accommodate the heterogeneous relationship between a scalar response and functional covariates. The proposed model is motivated by a salinity tolerance study of barley families, whose main objective is to detect salinity tolerant barley plants. Our model is flexible, allowing for heterogeneous functional coefficients while being efficient by pooling information within a group for estimation. We develop an algorithm in the spirit of the K-means clustering to identify latent groups of the subjects under study. We establish the consistency of the proposed estimator, derive the convergence rate and the asymptotic distribution, and develop inference procedures. We show by simulation studies that the proposed method has higher accuracy for recovering latent groups and for estimating the functional coefficients than existing methods. The analysis of the barley data shows that the proposed method can help identify groups of barley families with different salinity tolerant abilities.     

THE PHYSICAL AND CHEMICAL ENVIRONMENT OF THE DEVELOPING EMBRYO OF PINUS RESINOSA

The relationship between changes in soluble protein, hexose sugar, total lipid concentration, and osmotic potential occurring in gametophytic supernatant of Pinus resinosa Ait. during in vivo embryogenesis was measured. The effects of varying sucrose levels of culture medium on in vitro embryo and gametophyte development were examined. Increases in embryo volume, and fresh and dry weight of the female gametophyte during in vivo embryogenesis coincide with increasing levels of soluble protein, hexose sugar, and total lipid in the gametophytic supernatant. In contrast, osmotic potential of the supernatant increased only slightly between the zygote and proembryo stages of embryo development, and remained constant thereafter. Gametophytes plus embryos grown in vitro achieved dry weights approaching those of in ovulo gametophytes on media containing levels of sucrose up to 21%. Gametophytes on media with sucrose concentrations up to 21% also resembled normal in ovulo gametophytes in appearance. However, embryo development appeared to be suspended on treatment media containing from 9% to 21% sucrose, while embryos degenerated on media with constant sucrose levels of 3% and 6%. A treatment medium containing approximately 12% sucrose would provide an osmotic environment that duplicates that found in ovulo. While greater sucrose levels promoted more normal gametophyte development in Pinus resinosa, we failed to achieve complete development of the embryo in vitro. Conclusions and implications drawn from these results are discussed.     

Respiratory function of velopharyngeal constrictor muscles during wakefulness in normal adults

Launois, Sandrine H., Judy Tsui, and J. Woodrow Weiss.Respiratory function of velopharyngeal constrictor muscles during wakefulness in normal adults. J. Appl.Physiol. 82(2): 584-591, 1997.The levator velipalatini (LVP) and the superior pharyngeal constrictor (SPC) influencevelopharyngeal patency and soft palate position, but their behaviorduring respiration is incompletely characterized. To further clarifytheir respiratory function, we recorded electromyographic activity(EMG) in the LVP and the SPC in awake normal subjects breathing orally.EMG data were obtained in six subjects for the LVP and in nine subjectsfor the SPC. EMG activity and timing and ventilation were measuredduring isocapnic hypoxia and hyperoxic hypercapnia. Phasic EMG activitywas inconsistently present during unstimulated oral breathing. Timingof EMG phasic activity was variable for both muscles. Peak LVP activitywas mainly or exclusively expiratory in three of six subjects. Peak SPCactivity was mainly or exclusively expiratory in five of nine subjects.With chemostimulation, recruitment of phasic activity was observed inthe LVP in four of six subjects and in the SPC in five of ninesubjects. Tonic activity increased in four of six subjects for the LVPand in three of nine subjects for the SPC. However, the response wasalinear, and intersubject as well as breath-to-breath variability wassubstantial. In conclusion, LVP and SPC are characterized by the highinter- and intrasubject variability of EMG activity, timing ofactivation, and response to chemostimulation.

     

Amino acid concentrations and selected enzyme activities in rat auditory,olfactory, and visual systems

Homogenates of specific brain regions of three sensory systems (auditory, olfactory, and visual) were prepared from pigmented Long-Evans Hooded rats and assayed for amino acid concentrations and activities of glutaminase, aspartate aminotransferase (total, cytosolic, and, by difference, mitochondrial), malate dehydrogenase, lactate dehydrogenase, and choline acetyltransferase. Comparing the quantitative distributions among regions revealed significant correlations between AAT and aspartate, between glutaminase and glutamate, between glutamate and glutamine, and between AAT plus glutaminase, or glutaminase alone, and the sum of aspartate, glutamate, and GABA, suggesting a metabolic pathway involving the synthesis of a glutamate pool as precursor to aspartate and GABA. Of the inhibitory transmitter amino acids, GABA concentrations routinely exceeded those of glycine, but glycine concentrations were relatively high in brainstem auditory structures.     

Junction formation in aggregated embryonal carcinoma cells

Under the action of supplemental calcium, H6 mouse embryonal carcinoma cell aggregates undergo compaction, a morphological phenomenon similar to mouse embryonic compaction. Formation of various types of cell junctions, especially gap junctions, is associated with compaction of the embryo and we sought to analyze the pattern of junction formation during aggregation and compaction of H6 cells. At 24 hr of aggregation, gap junctions were abundant in both uncompacted and compacted aggregates but quantitative analysis of freeze fracture replicas of these junctions showed a 20-fold increase in the size of the largest gap junctions in compacted aggregates. Such a difference in size could even be detected at 12 hr of aggregation. Tight junctions were not normally formed in 12 hr aggregates but initial stages of tight junction formation could be noticed in 12 hr compacted aggregates. More definitive tight junctions and desmosomes were evident only after 48 hr of aggregation. Thus we have observed that both uncompacted and compacted aggregates can form gap junctions at similar frequencies, suggesting that cell flattening, which contributes to the compacted morphology, is not a requisite for gap junctions. Likewise, generation of the compacted morphology seems to be independent of gap junction formation. This supports the idea that compaction in embryonal carcinoma cells results from calcium-induced cell flattening, probably through the mobilization of cytoskeletal elements. Calcium-dependent features of H6 cell aggregation and compaction enables the independent analysis of separate steps in compaction.