Constancy of DNA content in adrenal medulla nuclei of cold-treated rats.

Reports of changes in DNA content of certain types of cells following exposure to conditions of stress has led to the suggestion that two kinds of DNA may be present. One is genetic DNA, and the other is called "metabolic" DNA. In a further attempt to investigate the possibility of this phenomenon, determinations of DNA content were made on Feulgen-stained nuclei of adrenal glands and kidneys in cold-treated rats. Feulgen-stained nuclei were measured by two-wavelength microspectrophotometry. Particular attention was given to the handling of the smears in hydrolysis and staining. Mean values of Feulgen-DNA contents in a total of 720 nuclei demonstrated (a) a constancy of DNA content within 2% in individual nuclei both in adrenal medulla and kidney cortex, (b) no more than an average of 2% difference in DNA content between control and experimental nuclei, and (c) no more than an average of 1.5% difference in DNA content between normal kidney cortex nuclei and normal adrenal medulla nuclei. These results confirm the view that the more precise the measurement, the more accurately the constancy rule is obeyed. Moreover, there is no support for the concept of a metabolic DNA in the rat adrenal medulla.

Body image, mood and quality of life in young burn survivors.

This study looks at the body image, mood and quality of life of a group of 36 young people aged between 11 and 19 years who had burns as children, compared with an age-matched control group of 41 young people who had not had these injuries. Participants completed the Body Esteem Scale (BES), the Satisfaction With Appearance Scale (SWAP), the Beck Depression Inventory-II (BDI-II) and the Youth Quality of Life Questionnaire (YQOL). It was hypothesised that young burn survivors would report more dissatisfaction with their appearance, a lower mood and a lower quality of life compared with non-injured controls. However, young burn survivors reported significantly more positive evaluations of how others view their appearance (p=0.018), more positive weight satisfaction (p=0.001) and a higher quality of life (p=0.005) than the control group. They also reported more positive general feelings about their appearance, although this was just below the level for statistical significance (p=0.067) and a similar mood to the school sample (p=0.824). The data suggest that young burn survivors appear to be coping well in comparison to their peers, and in some areas may be coping better, in spite of living with the physical, psychological and social consequences of burns.

Induction of a Pea Cell-Wall Invertase Gene by Wounding and Its Localized Expression in Phloem.

A full-length cell-wall invertase cDNA obtained from pea (Pisum sativum L.) seedlings was cloned previously. The gene exhibits tissue-specific expression, and both its transcript and enzyme activities are abundant only in root tissues. Mechanical wounding dramatically induced the accumulation of the cell-wall invertase mRNA in detached or intact leaves, stems, and roots. In both detached and intact tissues mRNA started to accumulate 3 h after wounding and in detached tissues (except root tissues) was much stronger and lasted longer compared with that in intact pea plants. The induction of cell-wall invertase by wounding was not systemic, since no significant increase of transcript was found in the unwounded tissues remote from the site of wounding. Accumulation of this cell-wall invertase was induced by abscisic or jasmonic acid, and in situ hybridization studies show that this invertase mRNA is differentially localized in wounded plant tissue, being most abundant in the phloem. mRNA accumulation was limited mainly to the wounded area; no significant increase was detected in the unwounded portions of the wounded stem segments. The results suggest that, as part of the wounding response, this cell-wall invertase may provide energy through hexose availability to companion cells in the phloem.

Acute effects of aminoglutethimide on testicular steroidogenesis in normal men.

Aminoglutethimide (AG), a known adrenal inhibitor, was administered acutely to normal men in order to study its effects on testicular steroidogenesis. Sixteen subjects between the ages of 21--30 yr received either placebo or 1250 mg AG in divided doses during a 24-h period. To reduce the contribution of adrenal steroids, adrenal function was inhibited in both groups of men by the administration of dexamethasone (2 mg) on the night of the experiment. As a result, mean morning plasma cortisol levels fell to less than 2 micrograms/100 ml. AG blunted the normal diurnal rise in testosterone, which was observed in men receiving placebo, and significantly suppressed the levels of this androgen at 0700 and 0900 h. Estradiol concentrations fell to a greater extent than those of testosterone. The differences between the placebo and drug treatment groups were significant at all time points except 1900 h. A pattern similar to that of estradiol was observed for plasma estrone. When the overall effect of AG administration was examined by analysis of variance, the differences in the levels of all three steroids produced by treatment were highly significant (P less than 0.01 to less than 0.001). After the inhibition of testosterone and estradiol levels, LH and FSH concentrations were significantly (P less than 0.01 and P less than 0.001, respectively) higher in men receiving AG than in those given placebo. However, the data exhibited a large variance due to pulsatile gonadotropin secretion. These observations suggested that AG induces an acute inhibition of testicular steroidogenesis and probably also of the aromatization of testosterone to estradiol.

Power spectral analysis and cortical coupling of EEG for young and old normal adults.

Power spectral analysis and cortical coupling were computed on eyes closed EEG recorded from 80 normal volunteers: 20 females and 20 males aged 25-35 years and 20 females and 20 males aged 55-70 years. Recordings were from Fz, Cz, C3, Pz and Oz areas. The results indicate that with increased age there was a greater uniformity of EEG activity across the brain. Magnitude of EEG power was significantly less variable across recording sites for the old than for the young. In addition, cortical coupling values were reliably higher for the older subjects indicating a greater congruity among EEG patterns. The EEG power and cortical coupling measures were significantly correlated and thus may indicate an underlying mechanism common to both. The results support a theory of decreased central inhibitory function in old age and may reflect an age-related breakdown of functional autonomy of cortical areas. Gender had no significant effect on either EEG power or cortical coupling.

3'-Amino-2',3'-dideoxyribonucleosides of some pyrimidines: synthesis and biological activities.

3'-Amino-2',3'-dideoxyribonucleosides of thymine, uracil, and 5-iodouracil (1-3) were synthesized from the corresponding 2'-deoxyribonucleosides via the threo-3'-chloro and the erythro-3'-azido derivatives. Corresponding aminonucleosides of 5-bromouracil, 5-chlorouracil, and 5-fluorouracil (4-6) were synthesized enzymatically with 3'-amino-2',3'-dideoxythymidine as the aminopentosyl donor and thymidine phosphorylase (EC 2.4.2.4) as the catalyst. 3'-Amino-2',3'-dideoxycytidine (7) was synthesized by amination of the 3'-azido precursor of 3'-amino-2',3'-dideoxyuridine. The biological activity of 3'-amino-2',3'-dideoxy-5-fluorouridine (6) was notable among this group of aminonucleosides. It had an ED50 of 10 microM against adenovirus and was not appreciably cytotoxic to mammalian cells in culture. It also had activity against some Gram-positive bacteria but not against a variety of Gram-negative bacteria. The other aminonucleosides (1-5 and 7) lacked or exhibited weak antiviral and antibacterial activities. The only compounds in this group that were appreciably toxic to mammalian cells in culture were the thymidine and deoxycytidine analogues (1 and 7).

Synthesis and biological activity of an acyclic analogue of 5,6,7,8-tetrahydrofolic acid, N-[4-[[3-(2,4-diamino-1,6-dihydro-6-oxo-5- pyrimidinyl)propyl]amino]-benzoyl]-L-glutamic acid.

The synthesis and biological evaluation of N-[4-[[3-(2,4-diamino-1,6-dihydro-6-oxo-5-pyrimidinyl)propyl]amino]- benzoyl]-L-glutamic acid (1) (5-DACTHF, 543U76), an acyclic analogue of 5,6,7,8-tetrahydrofolic acid (THFA), are described. The key intermediate, hemiaminal 8, was prepared in four stages from 3-chloropropionaldehyde diethyl acetal. Reaction of 8 with dimethyl N-(4-aminobenzoyl)-L-glutamate gave the 2,4-bis(acetylamino) derivative 11, which was hydrolyzed with 1 N sodium hydroxide to give 1; the glycine analogue 16 was prepared in a similar manner. The N-methyl analogue 2 and N-formyl analogue 3 were prepared from 11 and 1, respectively. Compounds 1-3 inhibited growth of Detroit 98 and L cells in cell culture, with IC50s ranging from 2 to 0.018 microM. Cell culture toxicity reversal studies and enzyme inhibition tests showed that 1 was cytotoxic but not by the mechanism of the dihydrofolate reductase inhibitor aminopterin. Compound 1 and its polyglutamylated homologues inhibited glycinamide ribonucleotide transformylase (GAR-TFase) and aminoimidazole ribonucleotide transformylase (AICAR-TFase), the folate-dependent enzymes in de novo purine biosynthesis; and 1 was an effective substrate for mammalian folyl-polyglutamate synthetase. The compound inhibited (IC50 = 20 nM) the conversion of [14C]formate to [14C]-formylglycinamide ribonucleotide by MOLT-4 cells in culture. These data suggest that the site of action of 1 is inhibition of purine de novo biosynthesis. Moderate activity was observed against P388 leukemia in vivo.

Pyrazolo[3,4-d]pyrimidine ribonucleosides as anticoccidials. 2. Synthesis and activity of some nucleosides of 4-(alkylamino)-1H-pyrazolo[3, 4-d]pyrimidines.

A series of 4-(alkylamino)-1-beta-D-ribofuranosyl-1H-pyrazolo[3, 4-d]pyrimidines was synthesized by enzymatic and chemical methods. On the basis of the previous finding that 4-(alkylthio)-1-beta-D-ribofuranosyl-1H-pyrazolo[3,4-d]pyrimidines were effective anticoccidial agents, this series was examined for efficacy against Eimera tenella in chicks. The most active anticoccidial agent in the present study was the 4-cyclopentylamino derivative (8), which cleared chicks of the parasite at 200 ppm in the diet. Some members of this series were toxic to embryonic chick liver cells, mouse cells, and human cells in vitro. The 4-diethylamino derivative (16), which was not toxic in vitro, appeared to be toxic in chicks.

Imidazo[4,5-c]pyridines (3-deazapurines) and their nucleosides as immunosuppressive and antiinflammatory agents.

A variety of imidazo[4,5-c]pyridines (3-deazapurines) were synthesized. With use of these aglycons as pentosyl acceptors, the corresponding ribonucleosides and 2'-deoxyribonucleosides were prepared by an enzymatic method involving transfer of the pentosyl moiety from appropriate pyrimidine nucleosides. With most of the imidazo[4,5-c]pyridines, the products obtained from the enzyme-catalyzed reactions were pentosylated exclusively in the 1-position. However, some 3-pentosylation occurred with aglycons that had H or N3 in the 4-position. In addition to the 2'-deoxy congener of the ribonucleoside of 4-amino-1H-imidazo[4,5-c]pyridine, the 5'-deoxy and 2',5'-dideoxy congeners were synthesized. All of the aglycons and their nucleosides were tested for toxicity to mammalian cells in culture. None were markedly cytotoxic. These compounds were also evaluated for their ability to inhibit lymphocyte-mediated cytolysis in vitro. 3-Deazaadenosine (23) and its 2'-deoxy congener (38) were the most potent inhibitors (ED50 = 20 microM). In addition to these two in vitro tests, in vivo inhibition of the inflammatory response in the rat carregeenan pleurisy model was determined. 3-Deazaadenosine (23) was the most potent compound (ED50 = 3 mg/kg) in this in vivo test.

High-resolution elasticity imaging for tissue engineering.

An elasticity microscope provides high resolution images of tissue elasticity. With this instrument, it may be possible to monitor cell growth and tissue development in tissue engineering. To test this hypothesis, elasticity micrographs were obtained in two model systems commonly used for tissue engineering. In the first, strain images of a tissue-engineered smooth muscle sample clearly identified a several hundred micron thick cell layer from its supporting matrix. Because a one-dimensional mechanical model was appropriate for this system, strain images alone were sufficient to image the elastic properties. In contrast, a second system was investigated in which a simple one-dimensional mechanical model was inadequate. Uncultured collagen microspheres embedded in an otherwise homogeneous gel were imaged with the elasticity microscope. Strain images alone did not clearly depict the elastic properties of the hard spherical cell carriers. However, reconstructed elasticity images could differentiate the hard inclusion from the background gel. These results strongly suggest that the elasticity microscope may be a valuable tool for tissue engineering and other applications requiring the elastic properties of soft tissue at high spatial resolution (75 microm or less).

[Pharmacokinetics of propofol as an induction agents in adults]. / Pharmacocinétique du propofol comme agent d'induction chez l'adulte.

The disposition kinetics of Propofol have been determined in 7 patients (3 men, 4 women) receiving 2.5 mg/kg for induction of anaesthesia. Peripheral venous samples were collected up to 12 hours after injection of the drug. Data analysis indicates a 3 compartment model with a terminal half-life of 480 +/- 141 min, clearance of 0.0352 +/- 0.0014 l/min/kg and volume of distribution of 24.2 +/- 6.2 l/kg.

Immunocytochemical localization of proteins in differentiating tissues of Pisum sativum.

Although there may be documented morphological changes during development, it is obvious that important changes in protein content occur in a vascular plant during the several stages of differentiation. In the absence of the latter information, an approach has been established for the localization of antigenic proteins in developing tissues of Pisum sativum. Monoclonal antibodies were raised to proteins extracted from pea internode tissue, and employed for the localization of three proteins in tissue sections. One of the proteins has two polypeptide subunits with molecular weights of 25,000 and 40,000 daltons, and the antibody binds to both of them. The three monoclonal antibodies produce different patterns of cellular localization in the tissue sections, as visualized by indirect immunocytochemical labeling. In another series of analyses, quantitative and qualitative differences in the protein contents of apical shoot tissue and mature internode shoot tissue have been found. These studies were based on the use of Western blots with both polyclonal (rabbit) antibodies and monoclonal (mouse) antibodies.

Rotational relaxation times of 1,6-diphenyl-1,3,5-hexatriene in phospholipids isolated from LM cell membranes. Effects of phospholipid polar head-group and fatty acid composition.

Phospholipids were isolated from mitochondrial, microsomal, and plasma membranes of LM cells and fractionated into individual phospholipid classes on silicic acid columns. The fatty acid composition and the rotational relaxation time of 1,6-diphenyl-1,3,5-hexatriene (DPH) were determined for each phospholipid class. Sphingomyelin was the only phospholipid isolated from LM cell membranes that showed a phase transition within the temperature range investigated, 5-40 degrees C. The rotational relaxation times for DPH were lowest in phosphatidylcholine in all the membrane fractions. Phosphatidylcholine isolated from the three membrane fractions of choline-supplemented cells had similar rotational relaxation times and phosphatidylcholine isolated from microsomal membranes of linoleate-supplemented cells had lower rotational relaxation times. The results indicate that the differences in the rotational relaxation times of DPH between mitochondrial, microsomal, and plasma membrane phospholipids could be explained primarily by differences in the polar head-group composition, while differences in the fatty acid composition had only a minor effect. This provides a basis for understanding how the different lipid components in these cells contribute to membrane fluidity.

The effect of age, sex and interstimulus interval on augmenting and reducing of occipital VEPs.

Three intensities of patterned and unpatterned flashes were used to elicit occipital visually evoked potentials (VEPs) from 4 groups of 15 subjects: boys and girls aged 7-9 years and boys and 15 girls aged 13-15 years. Interstimulus intervals (ISIs) of 1, 2 and 4 sec were used. Amplitude/intensity (A/I) slope was computed for each ISI and flash type condition and an estimate was made of the ability to differentiate patterned from unpatterned stimuli. The latter was inferred from coefficients of correlation which reflected the degree of similarity between VEP wave forms elicited by the two types of flashes. Responses of younger children attenuated to increasing stimulus intensity (negative A/I slope), while VEP amplitude of older subjects augmented to brighter flashes. VEP amplitude reduction may be the response of an immature brain to overstimulation. Amplitude reducing by 7-9-year-old children does not appear to be related to insufficient time between flashes for visual system recovery, since ISI did not differentially affect A/I slope of the younger and older subjects. Longer ISIs did, however, tend to produce more positive A/I slopes for all subjects, but only for patterned stimuli, suggesting that pattern detectors do not fully recover with ISIs of 1 and 2 sec. The visual system of young children was relatively less efficient than that of older children at differentiating patterned from unpatterned stimuli, indicating a differential sensitivity of pattern detectors as a function of age. Gender was not a factor in either magnitude of A/I slope or similarity of VEP wave forms.

Age-related decrements in Stroop Color Test performance.

Administered a modified Stroop Color-Word Test to 80 healthy males aged 21-90 years as part of a study of neuropsychological functioning. No age differences were demonstrated on simple reading tasks; however, significant age effects were observed for the color naming and interference tasks (ps less than .001). Ss in the oldest age groups (61-70 and 71-90 years) performed more slowly than younger individuals on both of these. Of a variety of tests administered, performance on the Stroop Test was most affected by age. Because impairment on Stroop color naming and interference tests appears to be a concomitant of normal aging, caution should be exercised when one is interpreting Stroop test results from older patients with suspected cerebral dysfunction.

Quantitative changes in the degree of chromatin condensation during the cell cycle in differentiating Pisum sativum vascular tissue.

Fixed tissue from the base of the fourth internode above the cotyledons of Pisum sativum plants were Feulgen-stained for DNA. In squash preparations three cell types: xylem vessel elements, phloem fibres and phloem sieve tube elements were identified. Measurement of Feulgen absorbance values in 0.5 X 0.5 microns measuring points across each nucleus were obtained with a scanning cytophotometer. Condensed chromatin was defined as any measuring point having an absorbance value greater than 0.32. Calculating 'percent condensed chromatin' as the percentage of the total nuclear DNA contained in those densely stained points, the pattern of change in this parameter during interphase in each cell type was observed. A generally similar pattern occurs in all three cell types. Percent condensed chromatin decreased from about 45% to 12% during G1, increased rapidly to the end of S, decreased during G2 and the following G1 to increase again during the subsequent S phase.

Pattern reversal evoked potentials: age, sex and hemispheric asymmetry.

Binocular pattern reversal evoked potentials (PREPs) and EEG power were recorded bilaterally from occipital scalp of 50 boys and 50 girls aged 5-14. PREP latency was unaffected by either age or gender. Amplitudes, however, were consistently larger for girls than boys, with differences diminishing by adolescence. Significant electrophysiological asymmetries were also found; larger PREP amplitudes were measured at the right occipital electrode than the left for the two waves investigated. These asymmetries were apparent in both girls and boys across all ages, although they tended to diminish with maturation. EEG alpha asymmetry did not correlate with PREP amplitude asymmetry. PREPs elicited by half-field stimulation of 8 girls aged 9-10 years determined that 'paradoxical lateralization' of the half-field response was the same for both left and right half-fields, thus failing to account for the asymmetry of the full-field binocular response.

Amplitudes of visually evoked potentials to patterned stimuli: age and sex comparisons.

Electrophysiological age and sex differences in visual pattern responsivity were investigated. Pattern reversal evoked potentials (PREPs) and visual evoked potentials (VEPs) to patterned and unpatterned flashes were recorded from 20 normal subjects in each of 4 groups: young females and males aged 25-35 years and older females and males aged 55-70 years. PREP waves N70-P100 and P100-N150 from the older women were significantly larger than those from subjects in the other groups; mean amplitudes for the young females, young males and older males were not different. A similar effect, unusually large potentials for the older women, was obtained for VEPs, but only for VEPs elicited by patterned flashes and recorded from occipital scalp, i.e., an area overlying visual cortex which is sensitive to lines and edges. Our findings suggest that the visual system of older females is unusually responsive to patterned stimuli.