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S A Kempson

Publications and source records attributed to S A Kempson.

At least 37 records · Page 2Linked to original sources
PubMed1997

A light microscopic and ultrastructural examination of calcified dental tissues on horses: 4. Cement and the amelocemental junction.

Ultrastructural examinations showed the diameter of cement lacunae to be greater in infundibular cement than in peripheral cement of upper cheek teeth, which in turn was greater than in the peripheral cement of the lower cheek teeth. However, numbers of lacunae/unit area remained similar in these 3 dentinal region. Two types of cemental hypoplasia were found in equine cheek teeth. The first type was termed central infundibular cemental hypoplasia and was confined to the central region of infundibular cement. The cement adjacent to these frequently large defects was very porous and contained large vascular channels. In recently erupted cheek teeth, these central infundibular cemental defects were filled with connective tissue. The size of these cemental defects, the relationships of such defects with the occlusal surface and the degree of porosity of cement surrounding these defects may be important in the development of cemental caries. The second type of cemental defect was found at the amelodentinal junction of both peripheral and infundibular cement and was termed junctional cemental hypoplasia and appeared as spaces varying from focal, to long narrow defects along the amelocemental junction with the adjacent cement of normal appearance. Peripheral cement was deposited both directly, i.e. on unresorbed or resorbed enamel surfaces or indirectly, where the cement was separated from enamel by a thin calcified layer. The surface of unresorbed enamel had a pitted appearance, with the bases of the pits formed by enamel prisms and the pit walls by interprismatic enamel. The cemental surface of resorbed enamel contained depressions of variable shapes and sizes. These depressions which are believed to be caused by the resorption of enamel by odontoclasts were both focal and diffuse and were more marked on the cemental surface of infundibular as compared to peripheral enamel.

Animals↗
PubMed1996

Renal brush border membrane Na/Pi-cotransport: molecular aspects in PTH-dependent and dietary regulation.

Inorganic phosphate (Pi) is reabsorbed in renal proximal tubules in a sodium (Na)-dependent manner involving brush border Na/Pi-cotransporter(s). Regulation of renal Pi-reabsorption, such as by parathyroid hormone (PTH) and/or by dietary Pi-deprivation, involves alterations in the rate of Na/Pi-cotransport. Two structurally different Na/Pi-cotransporters have been identified: type I-transporter and type II-transporter. The related mRNAs and proteins are located in the proximal tubule and in the brush border membrane. In heterologous expression systems type I and type II Na/Pi-cotransporters mediate Na/Pi-cotransport. Characterization of the transport properties suggested that the type II transporter is "responsible' for brush border membrane Na/Pi-cotransport (as observed in isolated vesicles). Administration of PTH to rats resulted in an inhibition of brush border membrane Na/Pi-cotransport (vesicles) and in a reduced brush border membrane content of the type II transporter. Feeding low Pi-diets resulted in an up-regulation of Na/Pi-cotransport (vesicles) and of type II transporter content; only after a prolonged exposure to low Pi-diets (more than 4 hr) was an increase in specific mRNA content observed. Refeeding high Pi diets had the opposite effects on Na/Pi-cotransport activity and on type II transporter protein. It is currently the task of future experiments to define the specific mechanisms leading to protein-synthesis-independent (PTH, acute Pi-deprivation, Pi-refeeding) and to protein-synthesis-dependent (prolonged Pi-deprivation) regulation of the type II Na/Pi-cotransporter.

Animals↗
PubMed1996

Hypertonic activation and recovery of system A amino acid transport in renal MDCK cells.

Amino compounds are abundant within the renal inner medulla, but their possible role during hypertonic stress is not clear. Renal epithelial Madin-Darby canine kidney cells were used to examine the osmoregulation of system A transport, a major Na(+)-dependent process for neutral amino acid transport. System A activity was markedly increased after 6 h of hypertonic challenge, and intracellular alanine content increased more than twofold. The activation of system A was reversed after 24 h of hypertonic challenge. This downregulation was accompanied by the activation of betaine transport, as measured by gamma-aminobutyric acid uptake. Extracellular betaine prevented the early activation of system A. The hypertonic activation of system A was blocked by actinomycin D and cycloheximide. When cells were returned to isotonic medium after hypertonic activation, the recovery of system A transport also was partially inhibited by actinomycin D and puromycin. The results are consistent with the possibility that hypertonicity, by disrupting a repressor protein, leads to increased synthesis of a system A-related protein. The isotonic recovery may require synthesis of new repressor proteins.

Amino Acids↗
PubMed1995

Parathyroid hormone inhibits plasma membrane Pi transport without changing endocytic activity in opossum kidney cells.

Parathyroid hormone (PTH) inhibits Na(+)-dependent Pi uptake in renal epithelial cells from opossum kidney (OK). This requires an intact endocytic pathway, suggesting that one action of PTH may be to promote endocytic removal of Na+/Pi cotransporters from the cell membrane. The present study tested if PTH, at a dose that inhibited membrane Pi transport, also produced an increase in endocytic activity. Pi transport was measured in isolated plasma membrane vesicles. Endocytosis was measured by allowing cells to take up horseradish peroxidase (HRP) followed by assay of triton-sensitive (latent) HRP activity in subcellular fractions isolated by density gradient centrifugation. Incubation of OK cells with 10(-7) M PTH for 3 h decreased Na+/Pi cotransport by membrane vesicles to 328 +/- 54 pmol/mg/min compared to 448 +/- 67 pmol/mg/min (mean +/- S.E., P < 0.03) in controls. Latent HRP content of endosomal fractions was dependent on the time and temperature used to load cells with HRP and on the concentration of HRP. However, incubation of OK cells with 10(-7) M PTH for either 1 or 3 h produced no change in latent HRP activity. Thus the action of PTH on the Na+/Pi cotransporter in the plasma membrane of OK cells does not require a change in the rate of endocytosis.

Animals↗
PubMed1995

Protein synthesis in tissues cultured from the bovine hoof.

The mechanical strength of the bovine hoof depends on keratinization of cells in the germinal layers of the epidermis. Histological examination of hoof tissues in calves and young heifers has identified disturbances in this keratinization process which would result in ineffective hoof development and could precipitate lameness. Short-term culture of bovine hoof tissue was used to investigate epidermal keratinization. Cell function remains viable in these cultures. The rate of protein synthesis, measured by [35S]-methionine incorporation, continued for at least 3 h in culture. Radiolabelled proteins in tissue homogenates were separated by SDS-polyacrylamide gel electrophoresis and characterised by fluorography and were representative of the proteins found in hoof tissue. Three prominent radiolabelled bands were identified as keratins and actin by Western blotting. Immunohistochemistry showed that keratin was localised principally in the epidermal layers, and microautoradiography indicated that this was the major site of protein synthesis. Hoof tissues cultured under these conditions provide a useful system for studying the acute regulation of epidermal keratinization.

Actins↗
PubMed1995

Microtubule disruption stimulates system A transport in cultured vascular smooth muscle cells.

This study has focused on the possible influence of microtubules for the regulation of Na(+)-dependent system A neutral amino acid transport in A10 cells, a cultured cell line derived from rat aortic vascular smooth muscle. When microtubules were disrupted by incubating cells for 5 h in serum-free medium containing colchicine, nocodazole, or vinblastine, there was a twofold increase in system A transport (Vmax change). The dose for the disruption of microtubules by colchicine was similar to the dose required for the stimulation of system A. The time course showed that system A stimulation did not occur until widespread disruption of microtubules was established. The stimulation was specific for system A; there were no changes in glucose transport and Na(+)-dependent transport of phosphate and glutamate. Serum refeeding of quiescent cells from 2 days of serum starvation led to stimulation of system A, glucose, and phosphate transport. However, only system A was activated when colchicine was added to the serum-free medium. Addition of colchicine during serum refeeding had no additive effect for the stimulation of system A. The stimulation by both colchicine and serum was blocked by cycloheximide and actinomycin D. These findings suggest that microtubule disruption may activate system A gene expression.

Animals↗
PubMed1995

Parathyroid hormone action on phosphate transporter mRNA and protein in rat renal proximal tubules.

The inhibitory action of parathyroid hormone (PTH) on Pi reabsorption in the renal proximal tubule is accompanied by a specific decrease in Na-Pi cotransport at the apical brush-border membrane (BBM). It is not known whether this decrease represents decreased activity of Na-Pi cotransporters already present in the BBM or whether the number of cotransporters is decreased. The present study of the molecular mechanism of PTH action made use of a specific cDNA probe and antiserum to a rat renal Na-Pi cotransporter (NaPi-2). Three groups of rats were used: intact controls, chronically parathyroidectomized (PTX), and PTX rats treated acutely (2 h) with bovine PTH-(1--34). Na-Pi cotransport by isolated renal BBM vesicles was increased to 1,315 +/- 44 in PTX rats, compared with 721 +/- 94 pmol.mg-1.10 s-1 in controls (P < 0.002), and was returned to control levels by PTH. Western blots of these BBM showed that PTX caused a 2.8-fold increase in NaPi-2 protein content, which was reduced to control levels by PTH. Immunohistochemistry of perfusion-fixed kidneys showed NaPi-2-specific immunofluorescence exclusively in apical BBM of proximal tubules. Expression of NaPi-2 protein at these sites was increased in PTX rats and decreased after PTH treatment. Northern analysis of total RNA showed that the abundance of NaPi-2-specific mRNA was not changed by PTX but there was a small decrease in response to PTH. The data indicate that PTH regulation of renal Na-Pi cotransport is determined by changes in expression of NaPi-2 protein in the renal BBM.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Osmoregulation of neutral amino acid transport.

Neutral amino acids are important organic osmolytes found in most osmotically stressed cells. During prolonged hypertonic stress, accumulation of neutral amino acids contributes to the cell volume recovery and may protect cellular proteins against salt denaturation. Hypertonicity activates system A transport, a major neutral amino acid transporter in mammalian cells, by a microtubule-dependent mechanism. Similar to the regulation of amino acid efflux induced by hypotonic-swelling, influx via system A may be under the control of intracellular ionic strength. An osmotically sensitive repressor may negatively regulate the gene expression of a regulatory protein which activates the preexisting system A transporter protein. Hypertonically activated protein kinases may be involved in this osmoregulation of amino acid transport.

Amino Acids↗
PubMed1994

Influence of the beta-agonist salbutamol on claw horn lesions and walking soundness in finishing pigs.

The effects of feeding the beta-adrenergic agonist salbutamol to pigs at levels between 0.5 and 5.0 mg/kg feed for different periods were studied in six experiments. The feet and ability to walk of pigs weighing 30 to 100 kg were examined at intervals. False sand-crack, white-line and heel erosion lesions were classified as mild or severe, and corns and wall haemorrhages were also recorded. At each examination the pig was given a grade for the overall severity of its foot lesions. Salbutamol fed at 1.0 to 5.0 mg/kg feed, for as little as 21 to 28 days, increased the frequency (P < 0.05 to 0.001) and often the severity (P < 0.05) of the foot lesions, the higher doses tending to produce more severe lesions, and the overall foot grades deteriorated (P < 0.001). The effects on both sexes were similar. No changes were observed when 0.5 mg/kg was fed for 56 days. Despite the severity of many of the foot lesions, the pigs became lame in only one experiment. Electron microscopy indicated that salbutamol was interfering with horn production, but light microscopy revealed no changes in skin sections. These findings suggested that salbutamol was not directly affecting the function of keratinocytes. Supplementing the diet of the pigs with biotin and methionine did not delay, or prevent, the effects of salbutamol.

Albuterol↗
PubMed1994

Increase of Na/Pi-cotransport encoding mRNA in response to low Pi diet in rat kidney cortex.

Renal proximal Na/Pi-cotransport is increased in response to low dietary Pi intake. Recently, a cDNA (NaPi-2) related to the rat renal brush membrane Na/Pi-cotransporter has been cloned. In the present study, we used rats fed for 6 days with either a low Pi diet (LPD) or a high Pi diet (HPD), respectively. In parallel to an increased renal brush-border membrane Na/Pi-cotransport in LPD rats, there was also an increased content of NaPi-2 mRNA in renal cortex. After injection into Xenopus laevis oocytes, mRNA isolated from LPD rats induced a greater increase in Na/Pi-cotransport compared to mRNA from HPD rats. Hybrid depletion experiments suggested that mRNA-induced Na/Pi-cotransport is related to NaPi-2. We conclude that chronic Pi deprivation leads to an increased brush-border membrane Na/Pi-cotransport via an increase in the level of (NaPi-2) mRNA.

Animals↗
PubMed1994

A comparative study of single- and double-bundle ACL reconstructions in sheep.

Work in vitro has previously shown superior restoration of knee stability using a double-bundle anterior cruciate ligament (ACL) reconstruction compared with single bundles taken through the condyle or 'over the top'. This paper describes an animal study designed to compare the three ACL reconstructions in vivo, in order to collect data that could support the clinical use of a double-bundle reconstruction. The three methods were compared in three groups of eight sheep, the ovine ACL having a distinct double-bundle structure. Biomechanically matched polyester fibre implants were used, with 6 months in vivo. The three methods led to similar intra-articular fibrous tissue integration of the implants and no evidence of implant damage, and biomechanical testing found greater laxity than normal for all three groups. The double-bundle group, however, had more joint surface degeneration than the other groups. It was concluded that clinical use of the double-bundle reconstruction was not indicated by the results of this experiment, in view of the more complex surgery and lack of superior performance.

Animals↗
PubMed1994

Hypertonic upregulation of amino acid transport system A in vascular smooth muscle cells.

The A10 line of vascular smooth muscle cells has Na+ dependent transport systems for alanine, proline, and Pi, whereas uptake of leucine, myo-inositol and D-glucose is Na+ independent. When A10 cells were incubated for 4 h in medium made hypertonic by addition of sucrose, there was a marked increase in Na(+)-dependent transport of alanine and proline but no change in Na(+)-dependent Pi uptake or Na(+)-independent uptake of leucine and inositol. Intracellular alanine content was increased 61% by the hypertonic treatment. Other nonpenetrating solutes, such as cellobiose and mannitol, reproduced the effect of sucrose, but urea, a penetrating solute, did not. Studies with 2-(methylamino)-isobutyric acid revealed that the upregulation by hypertonicity involved only system A. Increases in alanine and proline uptake also occurred after incubating the cells in isotonic medium containing 0.1 mM ouabain, suggesting that an increase in intracellular Na+ may be part of the intracellular signal for upregulation of system A. Hypertonic upregulation of Na(+)-dependent alanine transport occurred also in primary cultures of vascular smooth muscle cells. The response was blocked by actinomycin D and cycloheximide, indicating that gene transcription and protein synthesis play important roles in the mechanism leading to increased alanine uptake. We conclude that vascular smooth muscle cells, during prolonged hypertonic stress, activate system A and accumulate specific neutral amino acids which may act as organic osmolytes to help maintain normal cell volume.

Alanine↗
PubMed1994

Vanadate action on renal phosphate transport.

Insulin stimulates reabsorption of phosphate (Pi) in the renal proximal tubule. Previous studies have shown that vanadate can mimic the action of insulin on various tissues. In the present study, we tested the action of vanadate on renal Pi transport both in control rats and in rats made diabetic by injection of streptozotocin. Vanadate was administered orally for 4 days by inclusion in drinking water (0.7 mg/ml). By the 4th day, vanadate treatment of control rats did not change acid-base status, plasma glucose or the filtered load of Pi, but the urinary excretion of Pi was reduced to 2.5 +/- 0.9 compared with 17.6 +/- 3.5 mumol/mg creatine (P < 0.02) in untreated control rats. However, Na+/Pi cotransport by isolated brush border membrane vesicles was not different between the two groups. Findings in parathyroidectomized rats were similar. By the 4th day of vanadate treatment of diabetic rats, there was reversal of polyuria, polydipsia and hyperglycemia with no change in acid-base status. The filtered load of Pi was decreased by vanadate, and urinary Pi excretion also tended to decrease but not significantly. The values for Pi excretion were 21.4 +/- 7.6 in vanadate treated diabetics and 36.1 +/- 4.5 mumol/mg creatinine in untreated diabetics. In contrast to vanadate, daily injections of insulin did not change the filtered load of Pi but reduced urinary Pi excretion in diabetic rats to 15.6 +/- 2.2 mumol/mg creatinine (P < 0.02). These findings suggest that vanadate stimulated tubular Pi reabsorption in control rats but not in diabetic rats. Vanadate treatment of diabetic rats may tend to decrease tubular Pi reabsorption in contrast to the action of insulin.

Acid-Base Equilibrium↗
PubMed1993

Phosphate deprivation inhibits NH4+ transport in OK cells.

Deprivation of dietary phosphate leads to a decrease in urinary excretion of ammonium in rats which may be mediated through an alteration in ammonium transport in the proximal tubule. In the present study the OK renal epithelial cell line, a model for the proximal tubule, was used to determine if NH4+ transport was changed during acute phosphate deprivation. The intracellular pH after perfusion with NH4Cl solution was used for calculation of intracellular NH4+ concentration. Intracellular [NH4+] increased linearly during the first 2 min of acidification with NH4Cl. NH4+ transport, defined by the initial rate of the increase in intracellular [NH4+], was decreased by 32% (P < 0.005) in phosphate-deprived cells. This transport process was inhibited by barium chloride, but not by DIDS, amiloride or ouabain, suggesting that the NH4+ transport pathway may utilize K+ channels. Acute phosphate deprivation may inhibit NH4+ transport in OK cells by decreasing membrane K+ permeability.

Ammonia↗