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L R Johnson

Publications and source records attributed to L R Johnson.

At least 109 records · Page 6Linked to original sources
PubMed1993

Transglutaminase in response to hypertonic NaCl-induced gastric mucosal injury in rats.

BACKGROUND: Polyamines serve as substitutes for transglutaminase-catalyzed protein cross-linking and are essential to the healing of gastric mucosal lesions. This study determines whether transglutaminase and protein cross-linking have a role in the healing of hypertonic NaCl-induced gastric lesions. METHODS: Rats were fasted 22 hours before given 1 mL 3.4 Mol/L NaCl intragastrically. Gastric mucosa was examined histologically and grossly, and transglutaminase activity was measured as the Ca(2+)-dependent covalent incorporation of [3H]putrescine into acid-precipitable protein. RESULTS: Transglutaminase activity increased significantly from 2 to 8 hours, peaking between 4 and 6 hours after NaCl administration. Lesions were significantly produced after 2 hours, and damage paralleled transglutaminase activity. Dansylcadaverine (200 mg/kg orally), a specific inhibitor of protein cross-linking, prevented the increases in transglutaminase activity and significantly delayed healing but had no effect on lesion formation. CONCLUSIONS: These results indicate that (1) hypertonic NaCl-induced gastric mucosal damage is associated with a significant increase in transglutaminase activity and (2) increased transglutaminase activity is involved in the mechanism of normal mucosal healing.

Animals↗
PubMed1993

Polyamines are necessary for cell migration by a small intestinal crypt cell line.

Studies from our laboratory have shown that polyamines are essential for the normal repair of duodenal erosions induced in vivo in a rat stress-ulcer model. In that model, the inhibition of ornithine decarboxylase, a rate-limiting enzyme of polyamine biosynthesis, with alpha-difluoromethylornithine (DFMO) almost entirely prevented healing. Healing could be restored by oral polyamines. In this paper, we have investigated whether the polyamines are required for the early stages of epithelial restitution using an IEC-6 cell culture model of cell migration. Treatment of the cells with DFMO for 4 days reduced cell migration 80%. Migration could be restored to normal by concomitant treatment with putrescine (PUT), spermidine (SPD), or spermine (SPM), but not by their addition during the migration period (6 h) only. If DFMO treatment was not begun until the migration period, it still reduced cell migration 20%, and this deficit could not be restored by concomitant addition of the polyamines. Intracellular polyamine levels at these times, i.e., 6 h or 4 days, were an important factor in these results. Only PUT was undetectable after 6 h of DFMO. SPD and SPM were still at normal levels at 6 h. SPD was undetectable at 4 days, but SPM was still at 40% of normal. These data give added importance to PUT because its absence reduced cell migration after only 6 h, while SPD and SPM were still present in normal amounts. Perhaps exogenous SPD and SPM restored cell migration when present with DFMO for 4 days treatment primarily because they contributed to intracellular PUT through the acetyltransferases.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Decreased expression of protooncogenes c-fos, c-myc, and c-jun following polyamine depletion in IEC-6 cells.

Direct exposure of small intestinal mucosal cells to luminal polyamines stimulates proliferation. This study tests the hypothesis that the protooncogenes c-fos, c-myc, c-jun, and junB are involved in the mechanism by which polyamines modulate mucosal growth. Studies were conducted in the IEC-6 cell line, derived from rat small intestinal crypt cells. Cells were grown in Dulbecco's minimal essential medium containing 5% dialyzed fetal bovine serum (dFBS) in the presence of absence of alpha-difluoromethylornithine (DFMO), a specific inhibitor of ornithine decarboxylase, which is the rate-limiting enzyme for polyamine synthesis. Cellular polyamine levels, cell growth, and relative abundance of c-fos, c-myc, c-jun, and junB mRNAs, were measured at 1, 2, 4, 6, 8, and 12 days after initial plating. The intracellular polyamines, spermidine and spermine, and their precursor, putrescine, in DFMO-treated cells decreased significantly at 2 days and remained depleted thereafter. Although DFMO profoundly decreased growth and final cell number, both control and DFMO-treated cells entered a plateau phase by 6 days. In control cells, c-myc and c-jun mRNA levels significantly increased on days 4-6 and then returned to a basal level of expression, which was maintained thereafter. c-fos mRNA in quiescent cells after 24 h serum deprivation was significantly stimulated by 5% dFBS, although a steady-state level of c-fos mRNA was undetectable in control cells. Treatment with DFMO not only prevented increased expression of c-myc and c-jun protooncogenes at 4 days, but also significantly reduced steady-state levels of c-myc and c-jun mRNA between 6 and 12 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Clinical pulmonary function testing in dogs and cats.

Pulmonary function testing is a relatively new area in clinical veterinary medicine, with few individuals currently obtaining TBFVL and pulmonary mechanics on a clinical cases of respiratory disease in dogs and cats. ABG analysis is a technique available to all veterinarians through commercial laboratories or local hospitals. Information obtained from these tests will allow for a better understanding of the pathophysiology of naturally occurring diseases as well as assisting in the determination of the efficacy of various therapeutic agents (such as bronchodilators), which to date has been lacking in veterinary medicine.

Animals↗
PubMed1992

Role of transglutaminase and protein cross-linking in the repair of mucosal stress erosions.

We have recently demonstrated that polyamines are absolutely required for gastric and duodenal mucosal repair after stress. Polyamines act as substrates for transglutaminase and facilitate protein cross-linking. The current study tests whether transglutaminase and protein cross-linking are involved in the mechanism of mucosal healing. Rats were fasted 22 h, placed in restraint cages, and immersed in water to the xiphoid process for 6 h. Animals were killed immediately or 4, 12, or 24 h after stress. Gastric and duodenal mucosa were examined histologically and grossly, and transglutaminase activity was measured. Transglutaminase activity in gastric and duodenal mucosa was increased significantly from 0 to 8 h, peaking 4 h after the 6-h stress period. By 12 h, enzyme activity in duodenal mucosa had returned to control values while gastric mucosal transglutaminase did not decrease to control values until 24 h. Mucosal recovery from lesions produced by stress was evident 12 h after stress and was almost complete by 24 h. Dansylcadaverine (100 mg/kg, orally), a specific inhibitor of protein cross-linking, not only prevented the increases in transglutaminase but significantly decreased healing in both tissues. Oral administration of the polyamine spermidine (100 mg/kg) immediately after stress totally prevented inhibition of repair caused by blocking ornithine decarboxylase with difluoromethylornithine (DFMO, 500 mg/kg). Administration of dansylcadaverine, together with spermidine, significantly prevented the beneficial effect of spermidine on mucosal healing in the DFMO-treated animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Migration of IEC-6 cells: a model for mucosal healing.

Cell migration is the principal force behind the early restitution of erosions of the mucosa of the gastrointestinal tract. Despite the importance of cell migration to healing, no attempts to study the process in culture have been reported. We have attempted to standardize conditions for migration and test the migration responses of the small intestinal epithelial crypt cell line IEC-6 in some experimental situations already well known in vivo. We found good correspondence between in culture and in vivo on the following points: 1) migration was independent of DNA synthesis; 2) DNA synthesis was not concentrated at the wound edge; and 3) inhibition of actin polymerization stopped migration altogether. In addition, the presence of an extracellular matrix maximized migration. Protein inhibitors with different modes of action inhibited cell migration to different degrees, not always commensurate with their inhibition of protein synthesis. Cell surface proteoglycans were important; hyaluronic acid had an effect, but the secretion of a migration-stimulating substance by wounded cells was equivocal. Significantly, alpha-difluoromethylornithine (DFMO), which inhibits ornithine decarboxylase and polyamine synthesis, almost totally prevented cell migration. Because DFMO also prevents healing of mucosal erosions in vivo, we believe that this model can be used, keeping in mind its spatial limitations, to study the process of cell migration involved in the early restitution of mucosal erosions.

Cell Communication↗
PubMed1992

Effect of putrescine on S-adenosylmethionine decarboxylase in a small intestinal crypt cell line.

Two key enzymes in polyamine biosynthesis are ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMDC). SAMDC decarboxylates S-adenosylmethionine, which then donates aminopropyl groups for spermidine and spermine synthesis. The purpose of our study was to determine whether putrescine, taken up from medium or synthesized endogenously by ODC, alters SAMDC activity. Studies were conducted in the IEC-6 cell line derived from rat small intestinal crypt cells. Cells were grown in Dulbecco's minimal essential medium containing 5% dialyzed fetal bovine serum (dFBS). They were deprived of serum for 24 h before experiments. Basal SAMDC activity was increased significantly by > or = 10(-4) M of putrescine. Lower doses had no significant effect. The same doses of putrescine decreased ODC activity to near zero. Asparagine at 10 mM or 5% dFBS not only stimulated ODC activity and the intracellular putrescine levels but also increased significantly SAMDC activity as well. ODC activity peaked at 3 h, and the maximum level of SAMDC occurred 3-4 h after exposure to asparagine or serum. Treatment with DL-alpha-difluoromethylornithine (DFMO), a specific ODC inhibitor, prevented the increases in both cellular putrescine levels and SAMDC activity in asparagine- and serum-treated cells. In the presence of DFMO, exogenous putrescine returned SAMDC activity toward control levels but had no effect on ODC. A very slight increase of SAMDC half-life in IEC-6 cells grown in the presence of putrescine was not statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosylmethionine Decarboxylase↗
PubMed1992

Circadian rhythm of ornithine decarboxylase activity in small intestine of fasted rats.

The aim of this study was to determine whether the circadian changes in ornithine decarboxylase (ODC) activity of different segments of the small intestine were governed by factors other than food intake. First, the effects of fasting on mucosal ODC activity were examined. The results indicate that mucosal ODC activity in 24 hr and 48 hr fasted rats decreased significantly compared with ad libitum-fed rats. Second, the circadian rhythm of mucosal ODC activity was characterized by measuring mucosal ODC activity in fasted rats at four time points (09:00, 15:00, 21:00, and 03:00 hr; light period: 06:00-18:00 hr). The results from this study indicate that there is a detectable baseline ODC activity in different segments of fasting intestine. In duodenum, mucosal ODC activity was highest at 15:00 hr (light period), a time at which the rat was normally not eating. In jejunum and ileum, mucosal ODC activity increased between 21:00 and 03:00 hr (dark period). The observation that small intestine exhibits a distinct circadian rhythm of ODC activity in fasted rats suggests that not only food but also intrinsic factors can modulate physiologic oscillations in mucosal ODC activity.

Animals↗
PubMed1992

Ornithine decarboxylase in large bowel mucosa: regulation by gastrin, secretin and EGF.

Rats were fasted 48 h and then injected once with either saline, pentagastrin, EGF, secretin or combinations of secretin and pentagastrin or EGF. Another group of rats was fasted and refed. Animals were killed 4 h later and ODC assayed in mucosa of the cecum, proximal colon, and distal colon. EGF significantly increased ODC activity in all 3 tissues. Secretin had no effect by itself on ODC or ODC stimulated by EGF. Pentagastrin significantly increased ODC of the cecum, and secretin completely inhibited the effect of pentagastrin. Refeeding fasted rats significantly induced activity in all three tissues. Immunocytochemistry using a highly specific polyclonal ODC antibody showed that ODC was confined to the crypt cells of the proximal colon. Antibody dilution techniques demonstrated that gastrin, EGF and refeeding increased the level of enzyme in these cells. Refeeding in addition caused the appearance of enzyme in surface epithelial cells. These results showed that colonic mucosal ODC is present in proliferative cells and is regulated by the same peptides known to regulate growth in this tissue. Colonic mucosal ODC also responds the same way as it does in the oxyntic gland and small bowel mucosa.

Animals↗
PubMed1992

Luminal polyamines substitute for tissue polyamines in duodenal mucosal repair after stress in rats.

The purpose of this study was to examine whether luminal polyamines administered exogenously accelerate the repair of stress-induced intestinal mucosal damage in rats. Rats were fasted for 22 hours, placed in restraint cages, and immersed in water to the xiphoid process for 6 hours. Animals were killed either immediately after the period of stress or at 4, 12, and 24 hours thereafter. Duodenal mucosa was examined histologically, and ornithine decarboxylase activity and polyamine levels were measured. Repair of duodenal mucosa after stress was extensively delayed by administering 500 mg/kg DL-alpha-difluoromethylornithine (DFMO) IP. DFMO also inhibited ornithine decarboxylase activity and prevented increases in duodenal mucosal polyamine content. Intragastric administration of the polyamines, putrescine, spermidine, and spermine (100 mg/kg), immediately after stress significantly prevented the decreased rate of repair caused by DFMO. Spermidine or spermine accelerated healing better than putrescine in the DFMO-treated rats. Spermine also significantly increased the normal rate of repair of stress-induced damage. The delayed recovery of mucosal DNA, RNA, and protein content following stress in the DFMO-treated rats was prevented by exogenous polyamines. The reduced levels of duodenal mucosal spermidine and spermine in stressed rats treated with DFMO returned toward control levels after administration of exogenous spermidine. These results indicate that (a) luminal polyamines effectively substitute for endogenously synthesized polyamines in the repair process of the duodenal mucosa, (b) luminal polyamines can increase the normal healing rate and, (c) polyamines accelerate healing by increasing both an early phase and a later phase dependent on cell renewal.

Animals↗
PubMed1991

Retrogasserian rhizotomy causes expression of nerve growth factor receptor-immunoreactive protein in motoneurons within the adult feline trigeminal motor nucleus.

The effect of lesions on nerve growth factor receptor (NGFr) immunoreactivity (IR) in motoneurons within the mature feline trigeminal motor nucleus was investigated. Ten days following complete unilateral retrogasserian trigeminal rhizotomy including transection of the trigeminal motor root, motoneurons within the ipsilateral trigeminal motor nucleus showed NGFr-IR. In contrast, motoneurons within the contralateral trigeminal motor nucleus and within both trigeminal motor nuclei in unoperated control subjects did not show NGFr-IR. It is suggested that the appearance of NGFr-IR in motoneurons within the ipsilateral trigeminal motor nucleus after rhizotomy represents an attempts towards recovery and may be associated with the regrowth of its cut axons.

Animals↗
PubMed1991

Putrescine uptake and release by a normal rat small intestine crypt cell line, IEC-6.

IEC-6 cells were cultured on permeable filter inserts with separate access to the apical and basolateral sides. [3H]Putrescine uptake favored the apical side and its release (in Earle's balanced salt solution containing 0.1% bovine serum albumin) was six times greater in the apical-to-basolateral than in the basolateral-to-apical direction. Release in DMEM did not show this preference. The uptake of [3H]putrescine was stimulated approximately 1.3 times the basal level by 10 mM asparagine (ASN) or 5% dialyzed fetal bovine serum whether the [3H]putrescine was added at a concentration of 1 or 100 nM. The increased uptake was maintained for up to 6 h. When [3H]putrescine was removed after 4 h of uptake, the cells continued to release it into the medium on both sides for up to 4 h. Stimulated cells released only 50% as much as unstimulated cells. Unlabeled putrescine reduced the uptake of [3H]putrescine with an IC50 of 1.81 x 10(-6) M (r = 0.9476) and 1.02 x 10(-6) M (r = 0.9967) for unstimulated and ASN-stimulated cells, respectively. When the intracellular putrescine was reduced by difluoromethylornithine, the uptake of [3H]-putrescine was not changed, but its release was inhibited. Sodium was not required for [3H]putrescine uptake or release. Although the stimulated cells attained intracellular levels of [3H]putrescine which, if expressed as concentration based on cell volume, were up to 500 times the original extracellular concentration, a true concentration gradient could not be proven because 85% of the [3H]putrescine was probably bound to polyanions as shown by butanol extraction.

Animals↗
PubMed1991

Polyamines and ornithine decarboxylase during repair of duodenal mucosa after stress in rats.

This investigation shows whether polyamines and ornithine decarboxylase have a role in duodenal mucosal repair following stress-induced microscopic damage. Rats were fasted for 22 hours, placed in restraint cages, and immersed in water to the xiphoid process for 6 hours. Animals were killed either immediately after the period of stress or at 2-hour intervals up to 24 hours thereafter. Duodenal mucosa was examined histologically, and ornithine decarboxylase and polyamine levels were measured. Ornithine decarboxylase activity was increased significantly up to 6 hours following stress, peaking at 4 hours at a level 10 times the prestress control. By 8 hours, enzyme activity had returned to near normal. Increases in mucosal putrescine, spermidine, and spermine content paralleled the changes in ornithine decarboxylase activity and peaked 4 hours after stress. Stress resulted in microscopic damage evidenced by a nearly complete absence of villi. Significant macroscopic lesions were not present following stress. Mucosal repair was evident 12 hours after stress and almost complete by 24 hours, although the restituted villi were short and blunted. The decreases in mucosal DNA, RNA, and protein content caused by stress were restored and reached near-normal levels 12 hours after the period of stress. In animals given the specific inhibitor of ornithine decarboxylase, alpha-difluoromethylornithine, increases in duodenal mucosal ornithine decarboxylase activity and polyamine levels were inhibited and mucosal repair was almost completely prevented following stress. alpha-Difluoromethylornithine also prevented the recovery of DNA, RNA, and protein content of the duodenal mucosa. These results indicate that duodenal mucosal damage following stress is repaired rapidly; the repair process is accompanied by significant increases in ornithine decarboxylase activity and polyamine levels; and the increases in ornithine decarboxylase and polyamines are absolutely required for the normal repair of the mucosa.

Animals↗
PubMed1991

Pulmonary health risks among northwest loggers.

Spirometry, respiratory symptom questionnaires, and chest radiographs were obtained from 688 loggers in Oregon and Washington. These were compared against previously published National Institute for Occupational Safety and Health studies of nonexposed blue-collar workers to determine if these predictions fit our population. The loggers forced expiratory volume in 1 second and forced vital capacity values were significantly greater than predicted, and their forced expiratory volume in 1 second/forced vital capacity values were less than predicted. The only consistent difference in symptoms between the sample and reference populations was for recent chest illnesses, which were more prevalent in the loggers than in the reference population. The chest radiographs showed a small excess of pleural thickening that we believe is most likely due to chest trauma. We conclude that the National Institute for Occupational Safety and Health studies spirometry prediction equations may not be generalized to other blue-collar populations.

Aged↗
PubMed1991

Mucosal ornithine decarboxylase, polyamines, and hyperplasia in infected intestine.

Ornithine decarboxylase (ODC), through the regulation of polyamine biosynthesis, influences normal mucosal growth and cell proliferation. The purpose of our study was to determine whether mucosal ODC activity and polyamines play a role in the dramatic increase in mucosal mass and crypt elongation associated with parasite-induced inflammation in the small intestine. Rats were inoculated orally with L1 larvae of the parasitic nematode Trichinella spiralis and killed at different times during the enteric phase of infection. Duodenal and jejunal mucosal ODC activities increased significantly from 2 to 14 days, peaking 7 days postinfection (PI). By 21 days PI, enzyme activity had returned to near normal values. In the ileal mucosa, ODC activity was increased only at 7 days PI. Increases in ODC activity were paralleled by increases in mucosal putrescine, spermidine, and spermine content. Infection with T. spiralis induced DNA synthesis and evoked a significant rise in DNA, RNA, and protein content in the mucosa. Increases in nucleic acid and protein levels were most prominent in the proximal half of the small intestine where the majority of worms reside. Treatment of rats with alpha-difluoromethylornithine (DFMO) prevented the infection-induced elevations in mucosal ODC activity, polyamine levels, DNA synthesis, and DNA, RNA, and protein content without influencing the development of inflammation or the parasite's life cycle. These results suggest that mucosal hyperplasia caused by infection may be regulated, in part, by the growth-promoting effects of ODC, presumably through the stimulation of crypt cell proliferation. Thus ODC may be an important determinant of the intestinal response to infection.

Animals↗
PubMed1991

Role of polyamines in gastrointestinal mucosal growth.

The polyamines have been under active investigation for nearly three decades. There is a great deal of evidence that they play an important role in gastrointestinal mucosal growth, but the mechanisms through which this role is carried out are still not fully explained. This review examines the role of the polyamines in the regulation of mucosal growth, the control of intracellular polyamine levels, the biosynthesis of the polyamines, and some known mechanisms of their action. Finally, we propose a model of polyamine action that reconciles the effects of various trophic agents and situations in which growth is stimulated along with concomitant changes in polyamine levels. It accounts for both humoral and gradient-oriented features of various adaptive responses of the gastrointestinal mucosa and is intended to provide a framework for the future investigation of the role of the polyamines in this tissue.

Animals↗
PubMed1991

Stimulation of proximal small intestinal mucosal growth by luminal polyamines.

The purpose of this study was to determine whether luminal polyamines stimulate intestinal mucosal growth in vivo. Rats received 2% alpha-difluoromethylornithine (DFMO) added to their drinking water throughout the experiment. The polyamines spermidine and spermine (3 mg each/100 g body wt) were given intragastrically in combined doses once at 9:30 A.M. and again at 5:30 P.M. Duodenal and jejunal mucosal ornithine decarboxylase (ODC) activity in the DFMO-treated rats was inhibited significantly for the duration of the study. DFMO also markedly decreased the rate of [3H]thymidine incorporation into DNA of duodenal and jejunal mucosa. The decrease in [3H]thymidine incorporation was significant 4 days and maximal 6 and 8 days after beginning treatment with DFMO. Decreased ODC activity and DNA synthesis were paralleled by decreases in total mucosal DNA, RNA, and protein content. Administration of the polyamines significantly reversed the effects of DFMO except the inhibition of ODC. In fact, there were no significant differences in mucosal growth parameters between the controls (without DFMO) and those treated with DFMO plus polyamines. Oral administration of spermidine and spermine at a dose of 4.5 mg each/100 g body wt for 6 days to rats not treated with DFMO increased the normal rate of mucosal growth in the duodenum and jejunum as well. Polyamine accumulation in IEC-6 cells was measured to determine whether it was altered by DFMO. IEC-6 cells took up [3H]putrescine and [3H]spermidine from their surrounding environment and the uptake was stimulated by serum. DFMO (5 mM) totally inhibited the increase in ODC activity but had no effect on the cellular uptake of polyamines in the presence of putrescine.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗