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P Grant

Publications and source records attributed to P Grant.

At least 19 recordsLinked to original sources

Spatio-temporal patterns of retinal ganglion cell death during Xenopus development.

During development of the retina in mammals and birds, most retinal ganglion cells (RGC) that are produced are eliminated later in development by cell death. In lower vertebrates, however, such massive cell death has not been observed; total ganglion cell number increases linearly during most of development. Using 3H-thymidine or 5-bromodeoxyuridine labeling of retinal cell nuclei, we have been able to identify postmitotic RGC populations in Xenopus central retina at different developmental stages and follow their fate during development to postmetamorphic stages. RGC populations that become postmitotic between embryonic stages 32 and 49, during the initial stages of retinal growth, lose 40-77% of their cells during metamorphosis (approximately 4,000-5,000 cells). Twenty percent of the RGC present at stage 54, which later disappear, represent the same population of dying RGC that were present at stage 49. This suggests that the ganglion cells that became postmitotic between stage 49 and 53/54 show no apparent decline in numbers during metamorphosis. Since thyroxine is known to stimulate an increase in RGC number as well as the extent of fiber projection on the tectum, we suggest that this reduction in RGC numbers is not due to thyroxine-induced neuronal cell death. After stage 54, however, binocular vision develops in Xenopus (Keating, '74) and ipsilateral fibers begin to grow into thalamic visual neuropils (Hoskins and Grobstein, '85). We suggest, therefore, that as in mammals, in which RGC elimination correlates with binocular segregation of contralateral and ipsilateral retinal axons in visual centers, a similar process may occur in the frog among those RGC projecting to thalamic visual neuropils.

Animals

Development of the tectum and diencephalon in relation to the time of arrival of the earliest optic fibres in Xenopus.

The development of the tectum and diencephalon in Xenopus has been investigated in relation to recent descriptions of the establishment of the retinotectal projection. Tritiated thymidine autoradiography and bromodeoxyuridine immunohistology were used to identify the stages at which cells became postmitotic. Cells in the diencephalon were found to become postmitotic before cells in the tectum. At the time of arrival of the first optic fibres (stage 37/38) no postmitotic cells appeared to be present in the tectal precursor region. The first postmitotic cells which could be definitely assigned to the tectum appeared between stages 41 and 45. The results suggest that the initial retinotopic ordering of optic fibres observed from stage 37/38 relates to the position of fibres in the optic tract and not the tectum.

Animals

Perception without attention: results of a new method.

Having found by the use of a new method for examining perception without attention that grouping and texture segregation do not seem to occur (see Mack, Tang, Tuma, Kahn, & Rock (1992) Cognitive Psychology, 24, we go on to ask what is perceived without attention using this new method. Our subjects receive only one inattention trial in a sequence of trials involving a visual distraction task. In addition to the distraction task in the inattention trial, subjects received a stimulus of which they had no prior knowledge or expectation and were questioned or tested directly afterward for their perception of that stimulus. Two subsequent trials containing test stimuli serve as within-subject controls. The results of a series of experiments indicate that the presence of one or more stimulus objects and their locations are preattentively perceived, as is their color, but shape is not. Because individual items are detected without attention, we conclude that perceptual organization is initially based on a principle in which connected regions of uniform stimulation are inferred to be discrete units (the principle of uniform connectedness). One striking, unexpected finding is that without attention many subjects have no awareness at all of the stimulus object, an effect we call inattentional blindness.

Attention

Inhibition of axonal development after injection of neurofilament antibodies into a Xenopus laevis embryo.

The ability to target specific cytoskeletal components in axons for disruption within intact developing embryos would provide a valuable tool for studying neuronal development. Neurofilaments are an attractive target for such an approach, because they are neuron specific and are expressed late in embryogenesis principally beginning during axon outgrowth. No pharmacological agents are currently available that disrupt neurofilaments without also affecting general development. One approach that has been used successfully to affect proteins in vivo is to inject specific antibodies into living cells. We employed this approach in Xenopus laevis embryos by injecting two antibodies directed against the middle molecular weight neurofilament protein (NF-M) into a single blastomere of a two-cell stage embryo. Injected antibodies could be detected for as long as 3.5 days in cells descended from the injected blastomere. Only cell bodies of neurons descended from anti-NF-M-injected blastomeres contained abnormal accumulations of intermediate filament proteins, and peripheral nerve development was unilaterally retarded in these neurofilament antibody-injected tadpoles. Such accumulations and peripheral nerve defects were not seen in neurons derived from uninjected blastomeres or from blastomeres injected with control antibodies. These data demonstrate the usefulness of specific antibodies to perturb neuronal development in intact frog embryos and, in addition, suggest a role for neurofilaments in axon elongation.

Animals

Principal neurofilament-associated protein kinase in squid axoplasm is related to casein kinase I.

A cytoskeletal extract of pure axoplasm, highly enriched with neurofilaments (ANF), was prepared from the giant axon of the squid. This ANF preparation also contained potent kinase activities which phosphorylated the Mr greater than 400,000 (high molecular weight) and Mr 220,000 squid neurofilament protein subunits. High salt (1 M) extraction of this ANF preparation solubilized most of the neurofilament proteins and kinase activities and gel filtration on an AcA 44 column separated these two components. The neurofilaments eluted in the void volume of the column while the kinase activities eluted in the 17-44-kDa range of the column. Two major kinase activities were measured in this peak of activity. One of these strongly phosphorylated the phosphate acceptor peptide Leu-Arg-Arg-Ala-Ser-Leu-Gly (Kemptide) and was completely inhibited by the selective inhibitor of cAMP-dependent kinase Thr-Thr-Tyr-Ala-Asp-Phe-Ile-Ala-Ser-Gly-Arg-Thr-Gly-Arg-Arg-Asn-Ala-Ile- NH2 (Wiptide). Since addition of cAMP did not stimulate activity, this suggested that this kinase was a free catalytic subunit of cAMP-dependent kinase associated with the neurofilaments. The second kinase activity most effectively phosphorylated alpha-casein, and this activity was not affected by Wiptide. The alpha-casein phosphorylating activity (ANF kinase) was the principal activity responsible for neurofilament protein phosphorylation, and was not inhibited by various inhibitors against second messenger regulated kinases, suggesting it was related to the casein kinase family. Four lines of evidence indicate ANF kinase was similar to casein kinase I. These were: 1) the apparent molecular weight determined by gel filtration and the chromatographic elution profile on phosphocellulose column corresponded to casein kinase I; 2) heparin, an inhibitor of casein kinase II at 2-5 micrograms/ml, stimulated both ANF kinase and purified casein kinase I at these concentrations, while CKI-7, a relatively selective inhibitor of casein kinase I, inhibited ANF kinase in a comparable dose-response fashion; 3) purified casein kinase I strongly phosphorylated both ANF protein subunits (like ANF kinase) whereas casein kinase II was relatively ineffective; and 4) tryptic peptide maps of the HMW and Mr 220,000 neurofilament proteins after phosphorylation by ANF kinase or purified casein kinase I showed similar 32P-peptide patterns.

Animals

Unusual variants of vaginal adenosis: a challenge for diagnosis and treatment.

Two unusual cases of vaginal adenosis in non-diethylstilbestrol (DES)-exposed patients are presented. These cases created an initial difficulty in histological classification and exclusion of the diagnosis of adenocarcinoma. The first case presented a problem of atypical columnar epithelium with simple gland architecture, while the second showed a pseudoinfiltrative pattern of small glands, but without cytological atypia. A diagnosis of glandular dysplasia (atypical columnar epithelium) was finally made in the first case and vaginal adenosis with unusual architectural features in the second. The first patient was treated by excision and the second expectantly. Subsquently, neither patient has developed carcinoma. The spectrum of glandular changes in vaginal adenosis appears analogous to that of the cervix. Until the natural history of sufficient numbers of these variants of vaginal adenosis have been studied, the analogous cervical condition may serve as a guide to prognosis. The diagnosis of invasive adenocarcinoma should be made cautiously unless there are both architectural and cytological features of malignancy.

Adenocarcinoma

Cigarette smoking: an independent risk factor for atherosclerosis in the hypogastric-cavernous arterial bed of men with arteriogenic impotence.

We investigated the relationship between cigarette smoking and atherosclerosis of the hypogastric-cavernous arterial bed by evaluating arteriograms of young impotent men referred for selective pudendal angiography. Those patients with hemodynamically significant atherosclerosis had smoked more pack-years than had patients without arterial disease. These differences were statistically significant (p less than 0.05) for the common penile artery (32.8 pack-years, 40 patients versus 22.3 pack-years 57 patients) and the dorsal artery (31.3 pack-years, 48 patients versus 22.0 pack-years, 49 patients). The effect of cigarette smoking as an independent risk factor for atherosclerotic disease in the hypogastric-cavernous arterial bed was evaluated as well. When controlled for age, trauma history, hypertension and diabetes, cigarette smoking was independently associated with atherosclerosis in the internal pudendal artery (p less than 0.05). The relative risk (and 95% confidence interval) of developing internal pudendal artery atherosclerosis for each 10 pack-years smoked was 1.31 (1.05 to 1.64). A third analysis investigated the potential interactive effects of cigarette smoking and pelvic or perineal trauma. A significantly higher incidence (p less than 0.05) of cavernous artery atherosclerosis was found among smokers with a history of chronic perineal trauma (33 patients) compared to nonsmokers with a similar history (25 patients). The findings of this study indicate that cigarette smoking is an independent risk factor in the development of atherosclerotic lesions in the internal pudendal and common penile arteries of young impotent men. Cigarette smoking appears to predispose these patients to early atherosclerotic lesions in the cavernous artery following chronic perineal trauma.

Adolescent

Relaxin and relaxin c-peptide levels in human reproductive tissues.

A radioimmunoassay for a representative portion of the c-peptide of human relaxins (H1 and H2) was developed and validated. Relaxin c-peptide is present in preprorelaxin and prorelaxin, and exists free after the maturation of relaxin is completed. The aim of the study was to identify possible production and storage sites of human relaxin by comparing c-peptide and relaxin levels in various human reproductive tissues. c-Peptide immunoreactivity was present in the corpus luteum, amniochorion, decidua and seminal plasma; this indicates that these tissues may be relaxin production sites. Relaxin was detected in the corpus luteum, amniochorion, decidua, trophoblast, seminal plasma, myometrium and fibroids. This suggests that these tissues are storage and/or target sites for relaxin. The highest concentration of relaxin was detected in decidua at term. This level decreased after labour. The concentrations of c-peptide and relaxin were approximately equimolar in serum during pregnancy. This study lends support to the concept that relaxin is produced and stored at extra-luteal sites; these sites of hormone production support a paracrine role for relaxin during pregnancy.

Amino Acid Sequence

Gynecomastia.

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Gynecomastia

Human relaxin. In vitro response of human and pig myometrium.

Human myometrial strips were excised at hysterectomy and cesarean section and allowed to contract spontaneously in a water bath. Porcine myometrial strips from midpregnancy were also collected. Recombinant human relaxin completely inhibited spontaneous myometrial activity in the pig at a concentration of 0.6 micrograms/mL, but human relaxin had much less of an effect or no effect on human myometrium at concentrations up to 7.5 micrograms/mL. Any effect of human relaxin on human myometrium was seen only in estrogen-primed human tissues; that effect was never greater than a 5% reduction in amplitude and 50% reduction in frequency. When contractions were stimulated with oxytocin or prostaglandin F2 alpha, relaxin had no inhibitory effect on either porcine or human myometrium at doses up to eight times the concentration of relaxin that had attenuated spontaneous contractions. Pretreatment with progesterone did not enhance the action of relaxin on human myometrium. The limited effect of human relaxin on human myometrium as compared to the marked inhibitory action of both porcine and human relaxin on porcine myometrial activity suggests that the species specificity does not lie with the relaxins but with the target tissues. Human relaxin H2 might not play a major role in the control of myometrial activity in the human.

Animals

Anti-anginal efficacy of intravenous amlodipine assessed by means of atrial pacing.

The anti-anginal efficacy of amlodipine was investigated in patients with angina pectoris by means of atrial pacing during cardiac catheterization. Intravenous administration of amlodipine significantly increased the time to pacing-induced angina and reduced the area of ST-segment change for an equivalent pacing rate, suggesting an improvement in this functional marker of ischaemia. In addition, a significant reduction in the double product suggests that amlodipine reduced myocardial oxygen consumption.

Amlodipine

Arteriogenic impotence: findings in 195 impotent men examined with selective internal pudendal angiography. Young Investigator's Award.

Selective internal pudendal angiography was performed in 195 men (average age, 35.4 years +/- 10.3) who were suspected of having arteriogenic impotence. In the majority of patients, disease was localized to the cavernosal arteries. A previous series that involved older patients had demonstrated significant disease in the hypogastric and internal pudendal arteries. When controlled for trauma, the data revealed no significant difference (X2 test, P greater than .10) in the distribution of hemodynamically significant penile arterial disease. However, in patients who had sustained major pelvic trauma, the common penile artery was frequently hemodynamically compromised. There is a great deal of variation in the origin of the internal pudendal artery. An accessory pudendal artery was demonstrated in 7% of the patients. If a selective internal pudendal artery injection fails to demonstrate the penile arterial anatomy, a less selective injection should be performed. Bilateral injections should always be performed, as unilateral arterial disease was present in 15% of the patients.

Adolescent

Pharmacodynamics of amlodipine: hemodynamic effects and antianginal efficacy after atrial pacing.

The hemodynamic effects and antianginal efficacy of 10 mg amlodipine administered intravenously were assessed for 45 minutes in 18 subjects with stable angina pectoris. After amlodipine the heart rate was increased from 75 +/- 12 beats/min to 80 +/- 15 beats/min (p less than 0.05) for at least 15 minutes, with a decrease in systemic vascular resistance of 1091 +/- 205 to 815 +/- 390 dynes/sec/cm5 and a decrease in mean arterial pressure at 30 minutes from 99 +/- 11 to 91 +/- 10 (p less than 0.05). There was no change in dp/dt or dp/dt/IP or in cardiac output, wedge pressure, or pulmonary artery pressure. In the parallel placebo group (n = 8) there was no change in any of the hemodynamic parameters. Time to pacing-induced angina was increased in the treated group (n = 12) from 6 +/- 3.2 minutes before the dose to 8.2 +/- 4 minutes after the dose (p less than 0.01) compared to the control subjects who were given saline solution, in whom the time increased from 7 +/- 1.5 minutes before the dose to 7.5 +/- 2.2 minutes after the dose (n = 9). The double product at an equivalent pacing time to the initial onset of angina was reduced after therapy from 15,590 +/- 1490 to 14,100 +/- 1193 with a reduction in ST segment shift from 11.9 +/- 9.4 mm2 to 6.2 +/- 5.6 mm2 (p less than 0.05). Amlodipine after intravenous use has a vasodilator effect and also increases the anginal threshold without deleterious negative inotropic effects.

Amlodipine