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Biomedical subjects

R Calvert

Publications and source records attributed to R Calvert.

At least 37 records · Page 2Linked to original sources

Developmental expression of two antigens associated with mouse intestinal crypts.

Two monoclonal antibodies were prepared against the duodenal mucosa of four-day-old mice (MIM-1/39 and MIM 1/130). The expression of the antigens was associated with the crypts of the small and large intestine in the fetus and adult. MIM-1/39 was present in epithelial cells of the intervillous areas in the small intestine at 17 and 18 days of gestation; afterwards its expression was detected only in crypt cells from birth to adulthood. Transition from the mouth of the crypts to intestinal villi was abrupt. Expression of MIM-1/39 was first detected at time of birth in the colon: In the adult, only crypt cells expressed the antigen and goblet cells were negative. Antigen MIM-1/130 was detected from 16 to 18 days of gestation in the small intestine, in the mesenchymal matrix lying under the intervillous epithelium. After birth, it was present in the pericryptal mesenchymal matrix. This antigen was also expressed at birth in the colon and remained in the pericryptal matrix in the adult. In vivo, multiple injections of an organic extract of rat amniotic fluid to mothers, starting at 14 days of gestation, induced a profound modulation in the pattern of expression of both antigens at 17 days of gestation: The pattern of expression was comparable to that observed at least 5 days after birth in untreated animals. The expression of both antigens before crypt appearance may reflect some molecular differentiation in preparation for the formation of crypts, while their association with differentiated crypts may indicate that they have a role in the maintenance of crypt functional and/or morphological integrity.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Sweat gland response to exercise in the heat among pre-, mid-, and late-pubertal boys.

Sweating rate (SR) of boys is lower than that of men. To assess the association between the response of individual sweat glands and physical growth and maturation, three groups of circumpubertal boys cycled at 50% VO2max in a climatic chamber (42 degrees C, 20% relative humidity). Based on Tanner staging (pubic hair), 16 were classified as prepubertal (PP, Tanner 1), 15 as midpubertal (MP, Tanner 2-4), and five as late-pubertal (LP, Tanner 5). Population density (PD) of the heat-activated sweat glands, the mean area of sweat drops (DA), and the proportion of skin covered by sweat (%A) were measured by skin photography and computer-assisted imaging analysis. Other measurements included rectal and skin temperatures (Tre, Tsk, respectively), heart rate (HR), and total body SR. The rise in HR, Tre and Tsk did not differ among groups. Whole body SR was significantly higher in the LP group compared with PP (PP = 4.95 +/- 0.23, MP = 5.79 +/- 0.20, LP = 6.70 +/- 0.42 ml.min-1.m-2) (mean +/- SEM). PD was significantly higher in the PP group (PP = 128 +/- 8, MP = 97 +/- 9, LP = 74 +/- 9 glands.cm-2), while DA was higher in the LP group (PP = 5.47 +/- 0.59, MP = 6.92 +/- 0.47, LP = 12.83 +/- 1.41 microns2.10(4)). %A did not differ among groups. The calculated SR per gland was higher among the LP groups compared with the less mature ones (PP = 4.6 +/- 0.3, MP = 7.2 +/- 0.8, LP = 9.6 +/- 1.0 nl.min-1).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Measuring compliance in methadone maintenance patients: use of a pharmacologic indicator to "estimate" methadone plasma levels.

A quantitative indicator of compliance is not available for methadone--the drug of choice for the treatment of opioid addiction. We successfully used low-dose phenobarbital (a valid pharmacologic indicator) to measure compliance by incorporating the drug into the methadone medication of patients attending an addiction unit. Plasma phenobarbital and methadone concentrations were measured in 20 (11 clinic-based and 9 community-based) patients receiving long-term treatment with the phenobarbital level-to-dose ratio, together with interviews, to validate methadone measurements and to monitor compliance. Patients attending the unit on a daily basis and who consumed their medication in the clinic were substantially more compliant than community-based patients. Laboratory measurements of phenobarbital and methadone helped to identify the use of illicit methadone, as well as incorrect self-administration, such as the consumption of several days' dosage at one time.

Adult↗

Sweat lactate in exercising children and adolescents of varying physical maturity.

This study attempts to explain some of the individual variability in sweating pattern by comparing prepubescents and pubescents. Sweating rate and muscular anaerobic capacity are higher in adults than in children; thus we hypothesized that sweat gland anaerobic metabolism, as reflected by lactate excretion, might be higher with advanced physical maturity (PM). Lactate concentration in sweat ([LAC]sw) was measured at various stages of PM in boys who exercised in the heat. The subjects were divided into three groups on the basis of Tanner staging: prepubertal (PP, n = 16), midpubertal (MP, n = 15), and late pubertal (LP, n = 5). Subjects cycled at 50% of maximal O2 uptake for three 20-min bouts, with 10-min rest periods, in 42 degrees C and 18% relative humidity. Sweat samples were harvested, and population density of activated sweat glands was determined after each exercise bout. [LAC]sw during bout 1 was higher in PP than in LP [PP = 22.2 +/- 2.2, MP = 19.5 +/- 1.4, LP = 14.3 +/- 1.3 (SE) mmol/l]. In all groups, [LAC]sw decreased during subsequent bouts, and there were no intergroup differences in [LAC]sw during bout 3 (PP = 11.2 +/- 0.4, MP = 10.6 +/- 0.5, LP = 9.7 +/- 0.2 mmol/l). [LAC]sw was inversely related to sweating rate. Lactate excretion rate per gland was greater with the increase in PM (PP = 61.0 +/- 8.2, MP = 79.1 +/- 11.3, LP = 99.9 +/- 11.0 pmol/min; P = 0.08).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Migration of fetal intestinal intervillous cells in neonatal mice.

The migration of intestinal intervillous epithelial cells labeled in the fetus was followed in neonatal mice. At 17 days of gestation, a first group of pregnant mice received three intraperitoneal injections of 3H-thymidine (150 microCi/injection) administered at 30 min intervals. Two mothers were sacrificed 3 hours after the first injection. Mice from different litters were also sacrificed on days 0, 2, 4, 8, 12, 14, and 16 after birth. A second group of pregnant mice was injected at 18 1/2 days of gestation and offspring were sacrificed on days 6, 8, 10, 12, 14, and 16 after birth. Segments of duodenum and ileum were fixed in glutaraldehyde, postfixed in osmium tetroxide, dehydrated, and embedded in Epon. Sections were stained with aldehyde fuchsin and processed for radioautography. By following the leading front and trailing edge of labeled cells in the longest villi of the duodenum and ileum, we observed that 1) extrusion zones become active immediately after birth and 2) the longest villi do not elongate until 10 days after birth in the duodenum and 14 days in the ileum, that is, when all labeled epithelial cells originally present in the fetus have been extruded. Moreover, by measuring the distance between the internal limit of the inner circular layer of smooth muscle and the intervillous epithelium at 17 days of gestation (12.95 +/- 1.18 microns) or the bottom of the crypts at day 3 (14.81 +/- 0.91 microns), we propose that crypts do not develop as downgrowths: rather the intervillous epithelium is reshaped and the crypt-villus junction moves upward, away from the muscularis externa.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The influence of uncertainty and premovement visual information on manual aiming.

Target-aiming studies in which premovement visual information is manipulated suggest that when vision is occluded, a brief visual representation of the target environment may be used to guide movement. The purpose of this work was to determine if the internal representation contains information about the whole movement environment or just specific information about the position of a single target goal. Two experiments were conducted in which we manipulated both target uncertainty and the visual information available before and during a target-aiming movement. Radial error differences between visual conditions and the independence of the vision and uncertainty manipulations support the hypothesis that subjects form a representation of the overall movement environment.

Adult↗

A visual representation and the control of manual aiming movements.

Three experiments were conducted to determine if a representation of the movement environment is functional in the organization and control of limb movements, when direct visual contact with the environment is prevented. In Experiment 1, a visual rearrangement procedure was employed to show that a representation of the environment that provides inaccurate information about the spatial location of a target can disrupt manual target aiming. In Experiment 2, we demonstrated that spatial information about the position of a target can be destroyed by a visual pattern mask, supporting our claim that the representation is visual. A target-cuing procedure was used in Experiment 3 to show that representation of target position can be useful for premovement organization in a target-aiming task. Together our findings suggest that a short-lived visual representation of the movement environment may serve a useful role in the organization and control of limb movements.

Journal Article↗

Microtubule-associated proteins MAP5 and MAP1x: closely related components of the neuronal cytoskeleton with different cytoplasmic distributions in the developing brain.

Monoclonal antibodies were used to explore the relationship between two similarly sized microtubule-associated proteins (MAPs), MAP1x and MAP5. Although the proteins detected by anti-MAP1x and anti-MAP5 co-migrate in SDS-polyacrylamide gels, the patterns of antigenic proteolytic fragments (epitope maps) derived from them were completely different. The results suggest either that MAP1x is more stable than MAP5 or that the MAP1x epitope is situated close to one end of the molecule and gives rise to a very short proteolytic fragment. Immunoprecipitation from brain supernatants with either antibody brought down protein that cross-reacted with the other antibody, indicating that individual molecules bearing both epitopes exist in brain. Peptide maps of the proteins immunoprecipitated with the two antibodies showed that they are closely similar. Despite these similarities, the two antibodies gave different staining patterns on sections of developing rat brain, anti-MAP5 staining both axons and dendrites whereas anti-MAP1x stained only axons. We conclude that the MAP5 and MAP1x molecules are very similar, and possibly identical. The difference in staining patterns with the two antibodies could be because there are two proteins present in brain, one in immature axons bearing both the MAP5 and MAP1x epitopes and another with a wider distribution bearing only the MAP5 epitope. Alternatively, there may be a single protein bearing both epitopes, with the MAP1x epitope being masked in neuronal dendrites and mature axons by covalent modification or inter-molecular binding.

Aging↗

Monoclonal antibody G10 against microtubule-associated protein 1x distinguishes between growing and regenerating axons.

Monoclonal antibody G10 binds to an epitope on a microtubule-associated protein and specifically labels elongating axons in the developing rat central nervous system. In this paper the expression of this microtubule-associated protein epitope has been studied in developing and regenerating rat peripheral neurons and sprouting and transplanted neurons in the central nervous system. During development bright immunofluorescent fibres were seen from embryonic day 12 (the earliest age studied) to birth, but immunoreactivity declined postnatally and was only barely detectable in adult sciatic nerves. In contrast to the situation in developing axons, immunostaining could not be demonstrated in axons regenerating after crushing of the sciatic nerve nor in axons growing from grafts of septal tissue into the adult hippocampus, despite good evidence for extensive reinnervation in both cases. Immunostaining was only seen at early stages within the septal grafts, and not as axons grew out to reinnervate the host. No staining was seen in areas of deafferented hippocampus where collateral sprouting takes place. These results suggest a molecular difference between the cytoskeletons of developing and regenerating or sprouting nerves.

Animals↗

Development of the cerebellum with particular reference to cellular differentiation in the external granular layer.

Immunocytochemical evidence of differentiation in developing human cerebellum is presented in this study. Antibodies to neuron specific enolase, neurofilament protein, glial fibrillary acidic protein, vimentin, cytokeratin, epithelial membrane antigen and lymphoid markers, DLC and Leu 7 were used. The external granular layer showed positivity with neuronal markers between 27 weeks gestation and 4 months postnatal, but was negative for all other markers including glial fibrillary acidic protein. Characteristic staining reactions were noted in the other cerebellar layers. Monoclonal antibodies, UJ13A (pan-neuroectodermal marker) and G10 (localising microtubule-associated protein MAP1x) were also used in a limited number of cryostat sections and were positive and negative, respectively, in the external granular layer. The results of this study are discussed in relation to the theory that the external granular layer may be one source of medulloblastomas.

Biomarkers↗

On the presence of intermediate cells in the small intestine.

In the small intestine, the presence of transitional cells or cells intermediate between Paneth cells and goblet cells has been reported frequently for 100 years. Light microscopy and, more recently, fine structural studies have indicated that secretory granules observed in intermediate cells share some morphologic characteristics with those of granular goblet cells and of Paneth cells. In order to verify if intermediate cells in the jejunum and ileum of the adult mouse have functional similarities with either granular goblet or Paneth cells, we have studied the incorporation of sulfur-35 by radioautography and the localization of lysozyme by immunocytochemistry. After radioautography, goblet cells and, to a lesser extent, granular goblet cells had incorporated sulfur-35, whereas Paneth cells and intermediate cells were completely negative. Immunolocalization of lysozyme was done by using rabbit anti-rat lysozyme and protein A-peroxidase. After demonstration of peroxidase activity, only Paneth cells were stained and intermediate cells were negative. Therefore, intermediate cells do not contain sulfomucin or lysozyme, and they are functionally different from goblet and Paneth cells. Their function remains unknown.

Animals↗

Disposition of ifosfamide in patients receiving ifosfamide infusion therapy for the treatment of cervical carcinoma.

The disposition of ifosfamide was investigated in nine patients receiving a total of 23 72-h infusions. There was no linear relationship between steady-state plasma concentrations and either vomiting or CNS toxicity. The steady-state plasma concentrations were reproducible within patients, but there was a wide variation between patients. The progress of the disease did not affect the disposition of ifosfamide.

Female↗

Hormonal regulation of epithelial cell proliferation in the fetal mouse duodenum in vitro.

In adult rodents, radioautographic studies have shown that cell proliferation in the small intestinal epithelium is confined to the crypts. However, before birth DNA synthesis occurs also on villi. The role of extrinsic factors in the establishment of the adult pattern of enterocyte proliferation was studied in 17-day fetal mouse duodenal explants cultured with a completely synthetic medium. The effect of dexamethasone (30 and 300 ng/ml), thyroxine (10 nM), and insulin (125 mU/ml) on labelling indices was determined after 48 hours of culture. At the onset of the culture period close to 60% of the cells located in the first five cell positions in the intervillous areas became labelled after 4 hours of culture in presence of 3H-thymidine; 8% of the other epithelial cells found higher in the intervillous areas and on villi were also labelled. After 48 hours of culture, well differentiated crypts have developed in all the explants cultured with or without the hormones. The addition of thyroxine or insulin did not modify the labelling indices. However, after 48 hours of culture, the presence of dexamethasone (both concentrations) significantly reduced the labelling index of the cells found over cell position 5 but had no effect on the labelling index of cells located at the bottom of the crypts. When labelling index was determined for each cell position in the crypts at the end of the culture period, it appeared that dexamethasone (300 ng/ml) had no effect thereon for the first five cell positions but had a significant influence on the transitional zone from cell position 6 through 15.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Low-dose phenobarbitone as an indicator of compliance with drug therapy.

1 To assess the potential value of low-dose phenobarbitone (PB) as a marker of compliance we studied the relationship between plasma level of PB and dose (2-16 mg daily) following 3 or 4 weeks treatment in healthy volunteers (n = 26) and in-patient volunteers (n = 7). 2 Also, to simulate poor compliance, PB levels were measured in some volunteers following alternate-day (n = 6) or short-term (n = 5) treatment with similar doses. These levels, expressed as the level: dose ratios (LDRs), did not overlap with those obtained following 3 or 4 weeks of daily PB intake. 3 To evaluate the efficacy of this marker in patients taking other drugs we gave a group of out-patients (n = 24) compound tablets containing B vitamins and a small dose (16 mg) of PB; their compliance over 2-5 weeks was assessed both by measuring plasma levels of PB and residual tablet counting. 4 In the latter study, as well as providing absolute evidence of good compliance by many patients, the plasma levels of PB proved particularly valuable when non-compliant individuals 'forgot' to bring their residual tablets. 5 We suggest that phenobarbitone, in doses low enough to be non-sedative and non-enzyme inducing, is potentially useful as a pharmacological indicator of compliance with drug therapy.

Adult↗

Alzheimer's paired helical filaments share epitopes with neurofilament side arms.

A panel of monoclonal antibodies to neurofilaments have been investigated with regard to the location of their respective epitopes on neurofilament polypeptides and their ability to label the neurofibrillary tangles and paired helical filaments (PHF) which are characteristic of Alzheimer's disease. All of the neurofilament monoclonal antibodies that label tangles and PHF are directed against epitopes in the side arm domains of the two larger neurofilament polypeptides, NF-H and NF-M, and do not recognise the alpha-helical rod domains of these proteins. Immuno-electron microscopy demonstrates that the neurofilament antibodies label the constituent PHF per se and do not simply stain neurofilaments that might be admixed with PHF. These neurofilament epitopes are differentially retained by PHF, following isolation. Thus, antibody labelling of PHF is not simply due to the presence of normal neurofilament polypeptides. We propose that in tangle-bearing neurons, neurofilaments are degraded by proteases and that it is fragments of the side arms which contribute to the composition of PHF.

Alzheimer Disease↗

Role of thyroxine and insulin on the development of the fetal mouse duodenum in organ culture.

The role of thyroxine and insulin in the regulation of proliferation and differentiation of the immature duodenal epithelium of the fetal mouse was investigated using an organ culture method with a serum-free medium. Thyroxine (10 nM) stimulates specifically the activity of maltase. Insulin (125 mU/mL) remains without effect on the maturation of all hydrolytic functions studied. Each hormone significantly increases the percentages of brush border enzyme activities liberated in the medium and reduces the amount of glucose released in the medium. In the presence of dexamethasone (76 nM) the effect of thyroxine on maltase activity is still observed. Finally, thyroxine and insulin do not modify the labelling index in the duodenal crypts of the explants in the presence or absence of dexamethasone. These findings indicate that thyroxine and insulin can act directly on the development of the fetal mouse duodenum at the end of gestation. Nevertheless, their implication in prenatal development of the gut functions appears to be of minor importance.

Animals↗

A microtubule-associated protein (MAP1) which is expressed at elevated levels during development of the rat cerebellum.

Monoclonal antibody (MAb) G10 labels a single high mol. wt. (HMW) band on Western blots of microtubule preparations from 2 day old rat brain. The G10 antigen is thermolabile and co-migrates with microtubule-associated protein (MAP)1 from young rat brain on low percentage (5%) polyacrylamide-SDS gels. The G10 antigen decreases by about five times from birth to adulthood in the rat cerebellum. The same single band is labelled on Western blots of homogenates of whole neonatal rat brain but no labelling is found using neonatal or adult kidney, lung or liver. We have therefore identified a brain-specific MAP1, designated MAP1(x). Immunofluorescence microscopy using MAb G10 on parasagittal sections of rat cerebella shows labelling of the newly formed molecular layer in 6 day old rats. Only a narrow band close to the pial surface is labelled in 18 day old animals, which disappears in the adult. Labelling of the cerebellar white matter found in young rats also disappears. Neurones but not flat cells in cerebellar cultures label with MAb G10. All staining patterns are consistent with an axonal distribution of the antigen. MAP1(x) may be part of a developmentally regulated microtubule structure.

Animals↗