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Biomedical subjects

R E Chapman

Publications and source records attributed to R E Chapman.

At least 19 recordsLinked to original sources
PubMed2003

Genetic analysis of spatial foraging patterns and resource sharing in bumble bee pollinators.

Conservation biologists, evolutionary ecologists and agricultural biologists require an improved understanding of how pollinators utilize space and share resources. Using microsatellite markers, we conducted a genetic analysis of space use and resource sharing at several spatial scales among workers of two ecologically dissimilar bumble bee species (Bombus terrestris and B. pascuorum) foraging in an urban landscape (London, UK). At fine scales, the relatedness of workers visiting small patches of flowers did not differ significantly from zero. Therefore, colonies shared flower patches randomly with other colonies, suggesting that worker scent-marks deterring visits to unrewarding flowers have not evolved as signals benefiting nestmates. To investigate space use at intermediate scales, we developed a program based on Thomas & Hill's maximum likelihood sibship reconstruction method to estimate the number of colonies utilizing single sites. The average number of colonies (95% confidence limits) sending workers to forage at sites of approximately 1 ha in area was 96 colonies (84-118) in B. terrestris and 66 colonies (61-76) in B. pascuorum. These values are surprisingly high and suggested that workers travelled far from their colonies to visit the sites. At the landscape scale, there was little or no genetic differentiation between sites. We conclude that urban habitats support large bumble bee populations and are potentially valuable in terms of bumble bee conservation. In addition, bumble bee-mediated gene flow in plants is likely to occur over large distances and plant-bumble bee conservation requires landscape-scale action.

Alleles↗
PubMed1997

Translational attenuation mediated by an mRNA intron.

BACKGROUND: The unfolded protein response (UPR) is an intracellular signaling pathway that is activated by the accumulation of unfolded proteins in the endoplasmic reticulum (ER). The UPR results in an increase in transcription of ER-resident proteins that facilitate protein folding in the ER. A key regulatory step in UPR activation is the regulated splicing of HAC1 mRNA, which encodes Hac1p, a transcription factor dedicated to this pathway. Hac1p can be detected only when the spliced form of HAC1 mRNA (termed HAC1i mRNA, for induced) is produced; this was surprising because the unspliced HAC1u mRNA (HAC1u for uninduced) is equally stable in cells. RESULTS: We show that in contrast to most other unspliced pre-mRNAs, the HAC1u mRNA is transported from the nucleus into the cytosol. Although HAC1u mRNA is associated with polyribosomes, no detectable Hac1pu is produced unless the intron is removed, indicating that the presence of the intron prevents mRNA translation. When it is produced, Hac1pu has a stability similar to that of Hac1pi, the form of the Hac1p that is produced from the spliced mRNA and that differs from Hac1pu by a short carboxy-terminal tail sequence. Hac1pu, however, is differently modified and less active in activating transcription. Interestingly, when transplanted into the 3' untranslated region of a completely unrelated mRNA, the HAC1 intron is sufficient to attenuate translation of the preceding open reading frame. CONCLUSIONS: We have shown that the HAC1 mRNA intron is both necessary and sufficient to prevent complete translation of polyribosome-associated mRNAs. To our knowledge, this identifies a new way by which translation of a mRNA can be attenuated.

Basic-Leucine Zipper Transcription Factors↗
PubMed1997

The unfolded protein response coordinates the production of endoplasmic reticulum protein and endoplasmic reticulum membrane.

The endoplasmic reticulum (ER) is a multifunctional organelle responsible for production of both lumenal and membrane components of secretory pathway compartments. Secretory proteins are folded, processed, and sorted in the ER lumen and lipid synthesis occurs on the ER membrane itself. In the yeast Saccharomyces cerevisiae, synthesis of ER components is highly regulated: the ER-resident proteins by the unfolded protein response and membrane lipid synthesis by the inositol response. We demonstrate that these two responses are intimately linked, forming different branches of the same pathway. Furthermore, we present evidence indicating that this coordinate regulation plays a role in ER biogenesis.

Endoplasmic Reticulum↗
PubMed1994

Vacuolar sorting. Tracking down an elusive receptor.

The receptor that sorts carboxypeptidase Y for transport to the vacuole in yeast has recently been identified; how vacuolar sorting is regulated is far from understood, but an intriguing model has been proposed.

Amino Acid Sequence↗
PubMed1994

The functioning of the yeast Golgi apparatus requires an ER protein encoded by ANP1, a member of a new family of genes affecting the secretory pathway.

Mnt1p is an alpha 1.2-mannosyltransferase which resides in an early compartment of the Saccharomyces cerevisiae Golgi apparatus. We have shown that the signal-anchor region is sufficient, and the transmembrane domain necessary, for its normal Golgi localization. This is similar to the transmembrane domain-mediated retention of mammalian glycosyltransferases, and distinct from the tail-mediated recycling retention of certain mammalian and yeast trans-Golgi proteins. To examine the mechanism involved in transmembrane domain-mediated retention, we have isolated six classes of mutants which fail to retain Mnt1p-reporter fusions in the early Golgi. These mutants all show additional phenotypes which are consistent with alterations in Golgi function. We have called the mutant classes 'gem', for Golgi enzyme maintenance. GEM3 is identical to the previously cloned gene ANP1, and homologous to VAN1 and MNN9. Together, these define a new class of proteins involved in the organization and functioning of the secretory pathway. Interestingly, Anp1p is localized to the endoplasmic reticulum (ER), implying that some function of the ER is required to maintain a functional Golgi apparatus.

Amino Acid Sequence↗
PubMed1994

Retrieval of TGN proteins from the cell surface requires endosomal acidification.

TGN38 is a protein of unknown function located in the trans-Golgi network (TGN) of mammalian cells. Its intracellular distribution is maintained by it being continuously retrieved from the plasma membrane. In this paper we show that when cells are treated with agents such as chloroquine which neutralize acidic organelles, the movement of TGN38 along the endocytic pathway is blocked. The same effect is observed with a second TGN protein, the protease furin. We show that the cytoplasmic tail of furin is sufficient to confer a chloroquine-sensitive TGN localization on a heterologous protein. These results imply that the internal pH of endosomes affects sorting processes mediated by signals in the cytoplasmic portion of proteins and have implications for the role of acidification in endosomal function.

Acids↗
PubMed1994

The effect of zinc deficiency on wool growth and skin and wool follicle histology of male Merino lambs.

The aims of this work were to quantify the requirements of Zn for wool growth in growing male Merino lambs, and to describe the histological lesions of Zn deficiency in skin and wool follicles. Four groups of male Merino lambs (n 4) weighing 22 kg were fed ad lib. for 96 d on diets that contained 4 (basal diet), 10, 17 or 27 mg Zn/kg. Sheep in a fifth group were fed on the diet containing 27 mg Zn/kg, but were pair-fed to sheep on the 4 mg Zn/kg diet. Zn was added to the basal diet as ZnSO4 to give the respective treatment concentrations. Sheep fed on the diet containing 4 mg Zn/kg showed clinical signs of Zn deficiency and lower feed intakes and wool growth than sheep in the other groups. Their wool fibres were improperly keratinized and the wool follicles contained a higher proportion of apoptotic bodies than other groups. There was no evidence of parakeratosis and the rate of bulb-cell production was not affected. Sheep from other groups showed no clinical signs of Zn deficiency, and mean feed intakes and growth rates did not differ significantly between sheep fed on diets containing 10, 17 or 27 mg Zn/kg. However, wool growth was reduced in sheep fed on the diet containing 10 mg Zn/kg compared with those fed on diets containing 17 or 27 mg/kg. The mean concentration of Zn in the plasma at which wool growth was 90% of maximum was 0.5 mg/l. The equivalent value for the diet was 12 mg/kg, with 95% confidence intervals of 8 to 16 mg/kg. The results suggest that Zn deficiency reduces wool growth through a specific mechanism, perhaps involving impaired protein synthesis.

Animals↗
PubMed1994

A comparison of stress in surgically and non-surgically mulesed sheep.

A comparison has been made in 9- to 10-month-old castrated male Merino sheep of the changes in plasma total cortisol concentration and behaviour after being treated by either the modified Mules operation or by topical application of a quaternary ammonium compound to achieve non-surgical mulesing. After surgical mulesing, plasma total cortisol concentration increased immediately and rapidly and reached a peak value in 15 minutes, whereas after non-surgical treatment an immediate rise did not occur, but a similar peak value was observed in blood samples collected 24 hours after treatment. The concentrations were lower in both groups at 48 hours. Likewise postural changes indicative of discomfort were immediately apparent in the surgically treated sheep, but not until 3 to 4 hours later in those treated non-surgically. Arena testing revealed that a lasting aversion to the person who restrained them during treatment developed in the surgically mulesed sheep, but not in those treated non-surgically. The non-surgical procedure did not create large open wounds, as did the surgical operation, but still achieved similar enlargement of the bare area on the breech, and healing was quicker in the non-surgically treated sheep.

Animals↗
PubMed1993

Positive identification in a case of intentional extreme fragmentation.

The investigation of the first of the murders of young males to which Jeffrey Dahmer confessed led to systematic survey of two acres of semirural property in Bath Township, Ohio. The survey revealed the fragmentary skeleton of a young adult male, as well as bones of several species of animals. Through archaeobiological analysis, the animal bones were identified and taphonomic modifications documented. The human bones were dry and weathered, and many were rodent-gnawed, indicating that they had been exposed for many years. The human bones displayed an extreme degree of splintering, warping, bending, and spiral breakage. The only relatively complete bone was a cervical vertebra. Dismemberment and breakage had been accomplished by cutting, blunt force, and, in the case of the ribs, manual bending. Even in so extreme a case of intentional fragmentation, however, forensic anthropological analysis resulted in positive identification, with the primary criteria based on matching osteological features evident in premortem and postmortem radiographs of a cervical vertebra.

Adolescent↗
PubMed1989

Characterization and localization of receptors for epidermal growth factor in ovine skin.

Specific receptor sites for murine epidermal growth factor (EGF) have been characterized and their distribution determined in ovine skin. Binding of 125I-labelled EGF to skin membrane particles was temperature- and time-dependent, with equilibrium being reached within 1 h at 23 degrees C. Analysis of skin biopsies collected from ten castrated Merino sheep demonstrated the presence of a single class of saturable, high-affinity binding sites with a dissociation constant of 64 +/- 4 (S.E.M.) pmol/l and a binding capacity of 33.8 +/- 4.5 fmol/mg protein. Skin particle binding of 125I-labelled EGF was inhibited equipotently by mouse salivary gland EGF, EGF produced by recombinant DNA procedures and urogastrone. The EGF peptides 1-48, 6-53 and 7-53, derived from the native molecule by enzymatic cleavage, were much less potent. The relative binding potency of these molecules was correlated with their ability to induce precocious eyelid opening in mice and to inhibit wool follicle activity. Synthetic fragments representing the major structural domains of the EGF molecule (EGF(29-44), EGF(33-42) and EGF(3-31] were inactive in both the receptor and bioassays. Autoradiography of skin sections incubated with 125I-labelled EGF in vitro or of sections from skin which was perfused with 125I-labelled EGF in vivo demonstrated that EGF receptors were localized in undifferentiated cells of the epidermis and sebaceous glands, the inner and outer root sheath and bulb of wool follicles and in dermal arterioles. Differences in receptor concentration were observed between follicles following in-vivo perfusion of 125I-labelled EGF but not when the in-vitro labelling technique was used. The presence of receptors in these regions is consistent with the morphological changes in sheep skin in response to EGF administration which have been reported previously.

Animals↗
PubMed1988

Effects of intradermally injected and topically applied mouse epidermal growth factor on wool growth, skin and wool follicles of merino sheep.

Twice daily intradermal (ID) injections of mouse epidermal growth factor (mEGF) in sterile saline for 1-4 days into delineated areas of skin of Merino sheep produced dose-dependent changes in wool follicles and fibres, ranging from slight reduction in follicle bulb size and transient disturbance of cuticle formation on some fibres to the induction of catagen of follicles and shedding of fibres with distorted, tapered ends. Regeneration of follicles commenced by day 7. By contrast, ID injections of saline did not affect follicle activity. The epidermis became thicker and more parakeratotic after multiple injections of mEGF than after injection of saline, but was almost normal again by day 14. Persistent small increases in sebaceous gland size, additional to those induced by ID injections of saline, and delayed small increases in sweat gland size also occurred after multiple injections of mEGF. Daily topical applications of mEGF in 50% (v/v) aqueous propylene glycol 5 days each week for 4 weeks did not affect wool growth or the follicles and other skin components. The only effect observed, due to application of the aqueous propylene glycol, was an increase in the number of layers of cornified cells in the stratum corneum of the epidermis, with the cells arranged in clearly discernible stacks. The effects produced by ID injections of mEGF indicate that mEGF acts directly on the pilosebaceous and epidermal components of skin.

Administration, Topical↗
PubMed1987

Apoptosis in wool follicles during mouse epidermal growth factor (mEGF)-induced catagen regression.

Depilatory infusions of mouse epidermal growth factor (mEGF) induced regressive involution (catagen) in the wool follicles of Merino sheep. The follicles were examined by transmission electron microscopy prior to infusion and at intervals during catagen regression in order to determine the mechanism(s) involved in follicle involution. Cell deletion by apoptosis occurred in all cell types in the proximal region of catagen follicles between 12 h and 6 days after the beginning of infusion. Apoptosis also occurred in the basal layer of involuting sebaceous glands at 2 and 3 days, following earlier mEGF-induced proliferation. This process involved nuclear chromatin condensation and margination in single, scattered cells which subsequently fragmented and were ultimately phagocytosed and degraded by adjacent unaffected cells. It was concluded that during mEGF-induced catagen, wool follicle involution was accomplished largely through cell death and deletion by apoptosis.

Animals↗
PubMed1985

Effects of zinc deficiency on the wool growth, skin and wool follicles of pre-ruminant lambs.

Two groups of 1-month-old pre-ruminant lambs of similar mean liveweights were fed identical liquid milk-replacer diets except that the zinc contents were either 5 micrograms (deficient diet) or 32 micrograms per gram of dry matter (control diet). These diets were fed for 4 weeks, after which all the lambs received the control diet for 2 weeks. In the lambs fed the deficient diet plasma zinc concentration decreased markedly during the first 2 weeks and skin lesions developed around their mouths. Autophagic vacuoles also developed in most follicle bulbs along with a variety of defects in the wool fibres and progressive inhibition of wool growth. Food intake and liveweight increase were not significantly depressed until the third and fourth weeks of feeding the deficient diet. During this period the wool was shed from the zinc-deficient lambs as a result of the fibres being degraded and distorted within thickened outer root sheaths in the distal (upper) parts of the follicles. In addition, the epidermis of the wool-bearing skin became slightly acanthotic and hyperkeratotic, although not parakeratotic. When the deficient lambs were fed the control diet for 2 weeks, their food intake, liveweight gain and plasma zinc concentration increased to almost those of the control lambs, but their rate of wool growth was still low and the epidermis had not returned to normal. Compared with previous studies the findings of this study suggest that pre-ruminant lambs may be more susceptible to the effects of zinc deficiency than ruminant lambs.

Aging↗
PubMed1984

Ultrastructural changes in the oviductal epithelium of Merino ewes during the estrous cycle.

Isthmic and ampullary oviductal epithelia sampled from Merino ewes at days -1, 1, 3, and 10 of the estrous cycle (estrus = day 0) were studied by scanning and transmission electron microscopy after fixation by vascular perfusion. Secretory cells, ciliated cells, and lymphocytelike basal cells were observed in both isthmic and ampullary epithelium at all stages of the estrous cycle studied and their ultrastructural features were analyzed. Synthesis of lamellated secretory granules occurred in the ampullary secretory cells during the follicular and early luteal phases, and their contents were released by exocytosis into the oviductal lumen during the luteal phase. Granule release was associated with nucleated apical protrusion of these cells into the oviductal lumen. No such secretory activity was displayed by isthmic secretory cells even though a few cells contained nonlamellated granules. Apocrine release of apical vesicles and accompanying cytoplasmic material from apical protrusions of ciliated cells occurred in the isthmus around estrus but not in the ampulla. This unexpected feature has not previously been reported in any other mammal. Dendritic basal cells were distinguished in the lower part of the epithelium by their heterochromatic nuclei, electron-lucent cytoplasm, and lack of attachment zones. No migration of basal cells was observed, and their ultrastructural features were similar in the ampulla and isthmus and at all stages of the estrous cycle examined. The function of these lymphocytelike cells in the epithelium is uncertain, but the presence of phagocytic bodies and lysosomes in 20% of them may indicate a phagocytic role.

Animals↗
PubMed1983

Morphological changes in the skin and wool fibres of Merino sheep infused with mouse epidermal growth factor.

Intravenous infusion of 4.5-4.7 mg of mouse epidermal growth factor (mEGF) into nine castrated male Merino sheep for 26 h resulted in complete casting of the fleeces 6-8 days later. The morphological changes which occurred in the skin were studied in skin samples taken before infusion and at intervals between 1 h and 42 days after the infusion had begun. Wool fibres from the shed fleeces were examined with the scanning electron microscope. Increased cell proliferation occurred in the epidermis and sebaceous glands, whereas the wool follicles regressed. Transient dermal haemorrhages occurred during the first 3 h of infusion. The fibre and inner root sheath in the keratogenous zone of 30-40% of the follicles were partially disrupted within the first 6 h of mEGF infusion; catagen began in all follicle bulbs within 24 h. Fibre and inner root sheath production, although markedly reduced, continued in about 60% of follicles which had partially regressed, but production ceased in the remainder in which tapered ends formed on the fibres prior to shedding. Follicles began to regenerate asynchronously 4-8 days after the beginning of infusion and completed their development during the next 3 weeks. The follicle regression and fleece casting induced by mEGF infusion, and subsequent follicle regeneration were completed more rapidly than observed previously with other depilatory agents, and, except for prolonged epidermal thickening, there was no lasting cutaneous abnormality.

Animals↗
PubMed1982

Effects of experimentally induced fleece-rot on the structure of the skin of Merino sheep.

Four medium- and six strong-wool Merino sheep were exposed to continuous simulated rain (4 x 5 mm/h) for periods ranging from 3 to 9 days. In response, the sheep developed characteristics typical of fleece-rot lesions such as exudative encrustation and bacterial discoloration of the fleece. Skin samples were taken periodically for histological examination from the trunk region of the sheep prior to, during, and after wetting. An inflammatory response to wetting occurred in the upper layers of the dermis and this caused the release of cells which began invading the epidermis after only 6 h of wetting. An associated increased in vascular permeability was observed macroscopically following intravenous injection of Pontamine Blue dye. The incidence and extent of the inflammation increased during subsequent wetting while epidermal acanthosis and hyperkeratosis became increasingly evident. Epidermal thickness returned to pretreatment levels by 13 days after cessation of wetting when inflammatory regions were absent from most of the sheep. Cellular lysis occurred in the suprabulbar and keratogenous regions of a small proportion of follicles (1-13%) during wetting, resulting in the interruption of fibre growth and keratinization in these follicles and the occlusion of follicle hair canals with inner root sheath material ('plugs') which persisted after wetting ceased. These morphological changes in the epidermis, follicles and fleece are considered to occur as a consequence of the initial rapid inflammatory response to wetting during the developing of fleece-rot lesions.

Animals↗