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The present studies were undertaken to assess the effects of 5'-N-ethylcarboxamideadenosine (NECA), an adenosine analogue, on erythropoietin (Epo) production. NECA (0.05 and 0.1 mumol/kg i.v.) produced significant increases in serum Epo levels (368.8 +/- 56.1 and 384.6 +/- 45.9 mU/ml, respectively) in exhypoxic polycythemic mice after a four hour exposure to hypoxia when compared with hypoxia controls (133.2 +/- 18.2 mU/ml). The hypoxic kidney Epo levels were 46.4 +/- 13.4 mU/kg kidney which were significantly higher than normoxic kidney Ep levels (< 1.24 mU/kg kidney). Theophylline (20 mg/kg i.p.), an adenosine receptor antagonist, significantly inhibited the stimulatory effects of NECA on serum Epo levels. In vitro cultures of an Epo producing hepatocellular carcinoma (Hep3B) cell line with NECA (> or = 10(-6) M) for 20 hours under hypoxic conditions (1% O2) produced significant increases in medium levels of Epo when compared with hypoxia controls. Hepatocellular carcinoma cells treated with NECA at a concentration range of 10(-7) M to 5 x 10(-5) M for one hour in a hypoxic atmosphere also had significantly higher cAMP levels than that of hypoxia controls. Scatchard analyses of [3H]NECA binding to membrane preparations of hepatocellular carcinoma cells showed low affinity binding sites with a dissociation-constant (Kd) of 0.44 microM and a binding capacity of 863 fmol/mg protein. These findings suggest that the increase in Epo production in response to NECA under hypoxic conditions can be attributed, at least in part, to stimulation of adenosine A2 receptors which is coupled to adenylyl cyclase activation.

A procedure for the purification of a Peruvian isolate (C1) of sweet potato feathery mottle potyvirus (SPFMV) and infective RNA has been developed. The use of Hepes [N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid] buffer containing urea and sodium EDTA as a base for tissue extraction and virus suspension enabled good yields of virus (35-50 mg/100 g) to be obtained from Nicotiana benthamiana L. Domin. A short RNA isolation procedure yielded infectious RNA, from which ds cDNA of nearly genome size could be obtained. Sweet potato feathery mottle potyvirus, Purification, RNA isolation, cDNA synthesis.

The present studies were undertaken to characterize erythropoietin (Ep) production in an Ep-producing hepatocellular carcinoma (Hep3B) cell line. Hep3B cells which had been maintained in culture were transplanted under the renal capsule and subcutaneously in nude mice. The Hep3B xenograft doubling time is approximately 7 days. The mean hematocrit value of the Hep3B tumor-bearing nude mice was 33.2 +/- 1.1% (n = 8), which was significantly lower than that of control nongrafted nude mice (40.8 +/- 1.7%, n = 5). The Hep3B tumor-bearing nude mice showed significantly higher Ep levels in the sera (37.5 +/- 5.5 munits/ml, n = 8) than control nude mice (13.5 +/- 2.7 munits/ml, n = 5). Ep levels in the sera were correlated (R = 0.714) with the total Ep in the tumor extracts, whereas no Ep was detectable in any of the kidney extracts. On the other hand, an inverse linear relationship (R = -0.811) between the hematocrit values and Ep levels in the sera was demonstrated in the Hep3B tumor-bearing nude mice. The Hep3B cells recultured after growing in the nude mice were capable of enhancing Ep production in response to hypoxia, very similar to the original Hep3B cells which had been maintained in culture during the same time period. In addition, 15-methyl-prostaglandin E1 at a concentration range of 4-400 ng/ml produced significant increases in Ep secretion and cAMP accumulation in Hep3B cultures under hypoxic conditions (1% O2). The Ep produced by Hep3B cells expressed 3.7 times higher in vitro bioactivity than immunoactivity. The bioactivity of Hep3B Ep was completely neutralized by an antibody to highly purified human recombinant Ep. In contrast, the in vivo bioactivity of the Hep3B Ep was less than one tenth of its immunoactivity. These results indicate that the Hep3B tumor-bearing nude mice and the in vitro Hep3B culture system may provide a reproducible model system which should be useful for studies of the mechanism of Ep production.

Changes in O2 tension such as those associated with hypoxic ischemia or hyperoxia may potentially modulate purine nucleotide turnover and production of associated catabolites. We used an isolated perfused rat lung preparation to evaluate the effect of O2 tension on pulmonary uric acid production. Three O2 concentrations (21%, normoxia; 95%, hyperoxia; 0%, hypoxia) were utilized for both pulmonary ventilation and equilibration of recirculating perfusate. All gas mixtures contained 5% CO2 and were balanced with N2. We used Certified Virus Free Sprague-Dawley male rats weighting 250-300 g, four to five rats in each exposure regimen. After a 10-min equilibration period, we measured uric acid levels at 0 and 60 min in lung perfusate and at 60 min in lung tissue. After 60 min of ventilation/perfusion, we observed significant uric acid accumulation in both lung tissue (25-60%) and perfusate (8- to 10-fold) for all three O2 regimens. However, hypoxia produced substantially greater net uric acid concentrations (net = the difference between zero and 60 min) than either normoxia or hyperoxia (1.5-fold in lung tissue, and 2-fold in perfusate, respectively). The data suggest that pulmonary hypoxia results in greater purine catabolism leading to increased uric acid production. Vascular space uric acid, as measured in the recirculating perfusate, was proportional to lung weight changes (r = 0.99) with hypoxia exhibiting the greatest values, possibly reflecting a linkage between tissue perturbation and uric acid release. Thus, measurement of uric acid may serve as a useful marker of adenine nucleotide turnover and lung injury.

The present study was undertaken to assess the adenosine receptor regulation of erythropoietin (Ep) secretion in hepatocellular carcinoma cells (Hep3B) in response to hypoxia. In vitro cultures of Hep3B cells with N6-cyclohexyladenosine (CHA) at a concentration of 10(-5)M produced significant increases in cyclic AMP accumulation (142.43 +/- 13.31 pmole/10(6) cells) after 1 hr and Ep secretion (29.83 +/- 1.69 mU/ml) after 20 hr when compared with their respective hypoxia controls (cAMP: 3.05 +/- 0.26 pmole/10(6) cells, Ep: 19.41 +/- 1.41 mU/ml). The stimulatory effects of CHA on both Ep secretion and cyclic AMP accumulation were significantly inhibited by 8-phenyltheophylline (8-PT) at a concentration of 5 x 10(-7). No significant change in cell growth was observed at the CHA and 8-PT concentrations used in these experiments employing a spectrophotometric method. Incubation with 8-bromo cyclic AMP (10(-4)M) in response to hypoxia also produced a significant enhancement of Ep secretion (30.74 +/- 0.50 mU/ml) when compared with hypoxia controls (22.69 +/- 0.23 mU/ml), whereas no significant increase occurred in a normoxic atmosphere. CHA inhibited specific binding of [3H]5'-N-ethylcarboxamideadenosine (100 nM) to Hep3B cell membrane preparations in a dose-dependent manner in the displacement experiments and IC50 was 7.72 x 10(-6)M. These results indicate that Ep secretion is stimulated by adenosine membrane A2 receptors which are linked to adenylyl cyclase activation.

Hypoxia is the fundamental stimulus for erythropoietin (EP) production. It is clear that hypoxia increases erythropoietin messenger RNA in a renal cell, which leads to the production of increased amounts of erythropoietin in the kidney. Hypoxia also increases external messenger substances that amplify the effects of hypoxia and increases erythropoietin messenger RNA to further accelerate erythropoietin production. Some of these messenger substances are adenosine, eicosanoids, oxygen-derived metabolites, and beta-2 adrenergic agonists that are postulated to act through the activation of cell membrane receptors and are coupled to an increase in a G stimulatory protein which activates adenylate cyclase. This leads to increased production of cyclic adenosine monophosphate (AMP) for the production of key phosphoproteins that are involved in the biosynthesis/secretion of erythropoietin. This paper considers the physicochemical properties of human erythropoietin, pharmacologic agents that increase and decrease erythropoietin production/secretion, serum erythropoietin levels in normal human subjects and in patients with several types of anemia, and a model for the role of adenosine and other external messenger substances in erythropoietin biosynthesis/secretion.

This study was undertaken to determine the production of tumor necrosis factor (TNF) and interferon-gamma (IFN-gamma) by biological response modifier (BRM) in patients with renal cell carcinoma (RCC). Peripheral blood mononuclear cells (PBMC), which were donated from thirteen patients with RCC and five healthy controls, were cultured with streptococcal preparation, OK432, and/or macrophage-colony stimulating factor (M-CSF), and the TNF levels and IFN-gamma levels in the supernatant were evaluated. TNF activities were assayed by cytotoxicity to L929 cells and IFN-gamma activities were measured by inhibition of the cytopathic effects of sindbis virus on FL cells. The highest levels of TNF in the supernatant were 235.4 +/- 96.0 U/ml/1 x 10(4) cells in patients with renal cell carcinoma and 251.6 +/- 71.8 U/ml/1 x 10(4) cells in healthy controls, which were noted at 12 hours of incubation with the concentration of OK432 adjusted to 0.05 KE/ml. There was no statistically significant difference between the TNF activities induced by in vitro culture of PBMC obtained from patients with renal cell carcinoma and those from healthy controls. The production of TNF by in vitro culture of PBMC with OK432 of 0.05 KE/ml was augmented by adding 100 U/ml M-CSF especially at 48 and 72 hours of incubation, whereas M-CSF alone did not stimulated TNF production. The medium levels of IFN-gamma in six different cultures of PBMC with 0.05 KE/ml OK432 at 12, 24 and 48 hours of incubation were 1.15 +/- 0.34 U/ml/l x 10(4) cells, 2.23 +/- 0.93 U/ml/l x 10(4) cells, and 7.83 +/- 4.00 U/ml/l x 10(4) cells, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

This study reports the effects of cyclic adenosine 3'-5' monophosphate (cAMP) and hypoxia on erythropoietin biosynthesis in an erythropoietin-producing hepatocellular carcinoma cell line (Hep3B). Erythropoietin levels in the medium and cell extracts of low-density Hep3B cells after 20-hour incubation under hypoxic conditions (1% O2) were 25.33 +/- 1.50 mU/ml/10(7) cells and 3.60 +/- 0.50 mU/10(7) cells, respectively. These levels were significantly higher than in the respective normoxic controls (medium, 2.51 +/- 0.31 mU/ml/10(7) cells; cell extracts, undetectable [less than 0.31 mU/10(7) cells]). Cobalt also produced a significant increase in medium and cell erythropoietin levels. However, hypoxia and cobalt alone failed to produce an increase in cAMP accumulation in the cell cultures. Erythropoietin levels in the medium and cell extracts from cells exposed to 8-bromo cAMP (1 x 10(-4) mol/L) and forskolin (4 x 10(-6) mol/L) in a hypoxic atmosphere were significantly (p less than 0.05) higher than in the respective hypoxic controls. In addition, forskolin produced a significant (p less than 0.05) increase in cAMP accumulation (180 +/- 11.5 pmol/10(6) cells) under hypoxic conditions compared with the hypoxic controls (cAMP, 2.27 +/- 0.33 pmol/10(6) cells). These results suggest that cAMP elevation is not required in vitro in Hep3B cells for the increase in medium and cell levels of erythropoietin after hypoxia, but may be involved indirectly in erythropoietin biosynthesis, secretion, or both in vivo through some synergistic action with hypoxia.

Oxidant-mediated epithelial injury and repair processes may promote the development of pulmonary fibrosis. The authors examined this hypothesis by inducing oxidant injury in hamsters with intratracheally instilled mixtures of glucose, glucose oxidase (GO) and lactoperoxidase at weekly intervals. Solutions containing denatured GO (DE) served as a control treatment. One and six days after each treatment, anesthetized animals were sacrificed and lavaged, and their lungs and plasma were preserved for further study. Although DE-treatment consistently evoked a transient, neutrophil-rich inflammatory response, no significant biochemical or morphologic changes were detected at the ensuing 6-day time points. In contrast, repeated GO treatments prolonged inflammation and injured the alveolar epithelium, evidenced by significantly greater levels of neutrophils and macrophages in bronchoalveolar lavage fluid (BALF) and increased BALF levels of protein, beta-glucuronidase and lactic dehydrogenase activities. Active GO also altered BALF lymphocytes and monocytes, but no discernable pattern emerged. Fibrotic, consolidated parenchyma appeared after the second and third GO exposures, coinciding with increased levels of total collagen, prolyl hydroxylase activity, and anti-oxidant enzyme activities. Although alveolitis and type II cell hyperplasia were observed after the initial treatment, polyplike nodules covered by hyperplastic, undifferentiated epithelium were evident after the third treatment. After each exposure, GO-treated animals had larger volumes of parenchymal lesion than DE-treated hamsters. These data indicate that normal alveolar epithelial repair processes were greatly disrupted by repeated oxidant injury and suggest that repeated and/or continued epithelial injury may contribute to the development of pulmonary fibrosis.

The present studies were undertaken to assess the direct effects of N6-cyclohexyladenosine (CHA), a stable adenosine analogue, on erythropoietin (Ep) secretion in hepatocellular carcinoma cells (Hep 3B). Ep levels in the medium of low density Hep 3B cells treated with CHA in concentrations of 10(-5) and 5 x 10(-5) M for 20 h under hypoxic conditions (1% O2) were significantly higher than that of hypoxic controls. In addition, CHA at the same concentrations produced significant increases in adenosine 3',5'-cyclic monophosphate (cAMP) levels in Hep 3B cells after 1-h incubation under hypoxic conditions when compared with hypoxic controls. Dibutyryl cAMP (10(-5), 10(-4) M) also caused significant increases in Ep secretion when compared with control hypoxic cells. On the other hand, 8-phenyltheophylline, an adenosine receptor antagonist, significantly inhibited the stimulatory effects of CHA on both Ep secretion and cAMP accumulation in the Hep 3B cell cultures in response to hypoxia. These data suggest that Ep secretion may be regulated by adenosine receptor-coupled activation of adenylyl cyclase and the generation of cAMP.

Oxidant-induced inflammatory and morphologic changes evoked by intratracheally instilled glucose, active glucose oxidase, and lactoperoxidase in hamsters are described. Similar solutions containing heat-denatured (DE) glucose oxidase served as a control treatment. During the 7-week post-treatment period, animals were sacrificed and lavaged, and their lungs were preserved for later study. Although a neutrophil-rich alveolitis was observed initially in both groups, the leukocytic influx and mild morphologic changes in DE-treated animals resolved within 7 days. In contrast, glucose oxidase mixtures caused necrotic alveolar epithelium, fibrin-like exudate, and significantly higher neutrophil and protein levels in bronchoalveolar lavage fluid (BALF) after 1 day. At 4 days, type II cell hyperplasia was extensive and BALF levels of all inflammatory cell types were significantly greater than BALF counts in DE animals. By 7 days, BALF macrophage and lymphocyte levels remained significantly elevated, and epithelial hyperplasia coincided with marked increases in lung hydroxyproline and interstitial cells. BALF parameters suggested that inflammation induced by glucose oxidase resolved within 15 days, yet total hydroxyproline levels remained elevated at 15 days and epithelial lesions were still detectable at 29 days. Oxidant-induced inflammation and epithelial injury were transient and preceded moderate collagen deposition.

The present studies were undertaken to assess the effects of atrial natriuretic factor (ANF) on erythropoietin (Ep) secretion in Ep-producing renal carcinoma (RC) cells using a sensitive radioimmunoassay for Ep. Human ANF produced a significant dose-related increase in Ep secretion at concentrations of 10(-7) and 10(-6) M when compared with vehicle controls. ANF (greater than or equal to 10(-9) M) also significantly increased the intracellular guanosine 3',5'-cyclic monophosphate (cGMP) concentration after 5-min incubation with the RC cells. Scatchard analysis of the human 125I-labeled ANF binding data indicated that the RC cells contain a single class of binding sites with a dissociation constant (Kd) of 93 +/- 1 pM and a binding capacity of 2,190 +/- 750 sites/cell. Incubation of the RC cells with 8-bromo-cGMP in concentrations of 10(-7)-10(-5) M also produced a significant dose-related enhancement of Ep secretion. These findings suggest that the increase in Ep secretion in response to ANF can be attributed, at least in part, to activation of guanylate cyclase, which is coupled to specific ANF receptors on the RC cell.

Bleomycin (BLM)-induced lung fibrosis has been shown to be accompanied by the activation of poly(ADP-ribose) polymerase and depletion of nicotinamide adenine dinucleotide (NAD) in the lung. Niacin, a precursor of NAD, was used in the present study to investigate its possible ameliorating effect on BLM-induced pulmonary fibrosis in hamsters. Niacin (500 mg/kg IP) or saline (IP) was injected daily for 16 or 23 days. On day 3, hamsters were treated with BLM (7.5 U/5 mL/kg) or an equivalent volume of saline intratracheally. BLM alone significantly increased lung hydroxyproline levels, bronchoalveolar lavage fluid protein concentration, and various inflammatory cell counts in the lavage in both experiments. In addition, BLM alone elevated prolyl hydroxylase and poly(adenosine-5'-diphosphate [ADP]-ribose) polymerase activities in the 3-week study. Niacin treatment significantly decreased BLM-elevated lung hydroxyproline, prolyl hydroxylase, and poly(ADP-ribose) polymerase activities. Histopathology revealed that niacin treatment attenuated BLM-induced thickened alveolar septa, foci of fibrotic consolidation, and accumulations of inflammatory cells in the parenchyma and air spaces. The ability of niacin to attenuate BLM-induced lung fibrosis in hamsters suggests that it may have potential as an antifibrotic agent in humans.

Blood sugar and plasma free fatty acids (FFA), immunoreactive insulin (IRI), and growth hormone (GH) responses to a protein-glucose meal were determined in normal infants and those with marasmus and marasmic kwashiorkor. Among the normal subjects, fasting blood sugar (BS), peak BS and IRI, and the IRI/BS ratio tended to decrease as age increased. Peak IRI was at least 13muU/ml above fasting in 21 of 24 infants. Fasting GH levels were high 38.5 +/- 13.6 (SD) and 26.3 +/- 14.0 ng/ml, in the two youngest groups (under 1 year) and were comparable with those of the late newborn period. They were slightly lower, 20.8 +/- 22.1, in those 12.5-18.5 months of age. Suppressions of FFA and subsequent rebounds were in close temporal relation to BS and IRI peaks and lows, but not the GH levels. GH was promptly suppressed by the meal, and in most infants secondary elevations were seen. Untreated marasmic infants had normal or low BS, correspondingly normal or low IRI, markedly elevated FFA (1,821 +/- 588 muEq/liter), and GH levels comparable with those of the control subjects. There was some delay in BS elevation and disappearance and poor insulin release after the meal, with only two of nine having elevations of at least 13 muU /ml The BS elevations and IRI responses, however, were adequate to block FFA release. GH levels were poorly suppressed by the meal but some infants had further elevations, possibly in response to protein. After partial rehabilitation, fasting BS and FFA and BS elevations after the meal were normal. A slight improvement in insulin release was apparent. Fasting GH levels and responses to the meal were normal. Fasting, minimally treated children with marasmic kwashiorkor (MK) had normal or low BS, normal or low IRI, normal FFA, and probably normal GH levels. There was considerable delay in BS elevation, moderately delayed glucose disappearance, and very poor or unmeasurable insulin release after the test meal; FFA and GH were poorly suppressed. After partial rehabilitation, fasting BS was normal, FFA levels were (630 +/- 163 muEq/l), IRI was still low, BS elevations and disappearance improved. IRI responses modestly improved, and GH responses were normal.

Localization of gamma-seminoprotein (gamma-Sm) was examined using horseradish peroxidase-labeled anti-gamma-Sm antibody (Chugai Corp. Ltd., Tokyo, Japan) by the enzyme-labeled antibody method in paraffin embedded specimens of 18 benign prostatic hyperplasias and 32 untreated prostatic cancers. The level of serum gamma-Sm was also determined by enzyme immunoassay in 10 untreated patients with prostatic cancer and 18 with benign prostatic hyperplasia. Specific gamma-Sm staining was detected in prostatic glandular epithelial cells and prostatic secretion of all specimens of benign prostatic hyperplasias and a half of the specimens of prostatic cancers. Specific gamma-Sm staining was shown to correlate with histological differentiation of the prostatic cancer, but no correlation was found between specific gamma-Sm staining and the level of serum gamma-Sm.

Four groups of hamsters were assigned as saline + saline, taurine + saline (TS), saline + bleomycin (SB), and taurine + bleomycin (TB). The animals were treated with either saline or taurine (500 mg/kg ip) for 1 week and just prior to intratracheal instillation of bleomycin (7.5 units/kg) or saline on the eighth day. Thereafter, taurine administration was continued ip (250 mg/kg) and in drinking water (1%) for another 14 days. Bleomycin-induced increases in lung collagen were significantly inhibited in TB hamsters. Plasma taurine concentration in the TS group was significantly higher than that of the other groups. Lung lavage (bronchoalveolar lavage fluid) taurine in the SB group was significantly higher than the saline + saline and TS groups. Bronchoalveolar lavage fluid supernatant protein and acid phosphatase levels in the SB and TB groups were significantly increased over the saline + saline and TS groups. Although the total number of cells recovered in bronchoalveolar lavage fluid was not different among the four groups, there were significantly fewer neutrophils in the TB as compared with SB hamsters. Morphometric analysis revealed less than half as much lesion (diffuse mononuclear alveolitis and multifocal fibroplasia) in TB as compared with SB hamsters. Also, consolidated foci were less frequent and smaller in TB as compared with SB hamsters. Taurine may attenuate bleomycin-induced inflammation and fibrosis by scavenging reactive oxygen metabolites.

Reactive oxygen species (ROS) have been closely associated with a number of pathological disorders, including interstitial pulmonary fibrosis. While models of ROS-induced fibrosis offer advantages over chemically-induced fibrosis, the biochemical and morphological features of ROS-induced fibrosis have yet to be extensively documented. In this study, we evaluated the effect of initial ROS dose on lung injury and repair. Male hamsters received a single dose of glucose, glucose oxidase and lactoperoxidase via the intratracheal route. From 3 to 14 days post-treatment, a significant dose-related body weight loss was observed. There was a trend towards greater mortality with increasing dose. After 2 weeks, we noted significant, dose-related increases in lung levels of collagen, lipid peroxidation products, nucleic acids, and protein. Similarly, total lung catalase, lactic dehydrogenase and glutathione reductase activities were also elevated significantly above control values in a dose-related fashion. A concurrent, dose-dependent thickening of alveolar septa in ROS-treated lungs was composed of epithelial hyperplasia, hyperemia, edema and accumulations of interstitial fibers and macrophages. Interstitial and alveolar macrophages in ROS-induced lesions were enlarged and contained numerous primary and secondary lysosomes. These results demonstrate that, in the hamster lung, injury induced by enzyme-generated ROS can initiate dose-dependent fibroproliferative changes which eventuate into interstitial fibrosis.

The purpose of this study was to analyze the cellular and noncellular components of bronchoalveolar lavage fluid (BALF) at varying times during the development of pulmonary fibrosis induced by bleomycin. Hamsters were killed and lavaged in situ following the administration of a single intratracheal injection of 1 unit of bleomycin or an equivalent volume of sterile isotonic saline. The results show that the total cell counts in the BALF of bleomycin-treated hamsters, as compared with controls, were increased 7.7, 4.4, 2.4, 1.6, and 1.9-fold at 2, 4, 7, 14, and 21 days after treatment, respectively. The predominant cell types in the BALF of control animals were macrophages which constituted 84% of the total cells, followed by lymphocytes, 11%. The predominant cell types in the BALF of bleomycin-treated animals were polymorphonuclear leukocytes (PMN) which constituted 65% at two days and approximately 50% of the total at 4, 7, and 14 days; at 21 days macrophages were the predominant cell type constituting 50%, followed by lymphocytes at 30%. However, the total number of lymphocytes was not increased at 21 days compared to previous times. The noncellular protein content of BALF from bleomycin-treated hamsters, an index of pulmonary vascular permeability, was increased to 224, 559, 637, and 270% of control (2.7 mg/lung) at 2, 4, 7, and 14 days after treatment, respectively, and returned to control levels at 21 days. The acid phosphatase activity in the supernatant of BALF of bleomycin-treated animals was significantly increased to 181, 181, 199, 176, and 125% of control (258 units/lung) at 2, 4, 7, 14, and 21 days, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

In the present study, the effects of intratracheal administration of bleomycin have been examined on daily food intake and on plasma levels of glucose, cortisol, and insulin, and on lung levels of calmodulin, calcium, and collagen in hamsters. Since bleomycin treatment caused nutritional deficiency leading to loss of body weight, we have included pair-fed and control-fed as control groups in order to rule out the nutritional deficiency-related effects on these measurements. Bleomycin-treated animals showed a dramatic decrease in daily food intake and body weight as compared to control-fed animals. Bleomycin-treated animals were hyperglycemic when compared to nutritionally comparable pair-fed animals and had plasma glucose levels similar to those of control-fed animals. Plasma cortisol levels in bleomycin-treated and pair-fed animals showed a time-dependent increasing trend, whereas plasma insulin levels in both groups tended to decrease. The lung levels of calmodulin and calcium in bleomycin-treated animals were significantly increased when compared with the pair-fed or control-fed group. Bleomycin-treated animals had significantly higher levels of lung collagen than pair-fed or control-fed at 7 and 13 days after treatment. The lung collagen content in pair-fed animals was significantly less than that of control-fed at 13 days. It was concluded that a disturbance in carbohydrate metabolism and increased lung levels of calmodulin and calcium might be somehow involved in fibroproliferative changes of the lung in bleomycin-treated animals.

Serum gamma-seminoprotein (gamma-Sm) in patients with prostatic disease was determined by enzyme immunoassay. A total of 136 patients including 13 untreated and 40 treated patients with prostatic cancer, 45 patients with benign prostatic hyperplasia (BPH) and 38 patients with other urological diseases were analyzed. The mean +/- SD of serum gamma-Sm in the 13 patients with untreated prostatic cancer and the 45 patients with BPH was 31.7 +/- 46.1 and 3.7 +/- 6.6 ng/ml, respectively, there being a statistically significant difference between the two groups. All patients with untreated stage A or B prostatic cancer had a serum gamma-Sm of less than 4 ng/ml (cut off value). The mean level of serum gamma-Sm was 5.1 +/- 1.9 ng/ml for all patients with untreated stage C prostatic cancer; 66% of them had a value above the cut off value. However, it was 55.9 +/- 52.6 ng/ml in all patients with untreated stage D prostatic cancer; 87.5% of them had a value above the cut-off value. These results suggest that gamma-Sm may be a useful tumor marker in the management of patients with prostatic cancer.

Erythropoietin (Ep) levels in spent culture media of a Hep G2 human hepatoblastoma cell line were measured by radioimmunoassay (RIA), fetal mouse liver erythroid colony formation (FMLC), and the exhypoxic polycythemic mouse assay (EHPCMA). The Hep G2 cells at high density produced approximately 700 mU/ml Ep when measured with the RIA. On the other hand, the Ep levels when assayed in EHPCMA and FMLC were 50 and 2,600 mU/ml, respectively. The bioactivity in FMLC was completely neutralized by an antibody to purified human recombinant Ep, indicating that the erythropoietic activity in the Hep G2 spent culture medium was immunologically equivalent to Ep. Ep levels in the medium from low-density Hep G2 cells in 5% O2 and 1% O2 were 2.5- and 4-fold greater, respectively, than that of 20% O2. In contrast, hyperoxia (40% O2) significantly inhibited Ep production. A significant increase in Ep secretion was also observed when the cells were incubated with cobaltous chloride (2 X 10(-6) -2.5 X 10(-4) M). Tunicamycin (0.5 micrograms/ml), which inhibits N-linked glycosylation, significantly reduced the enhancement of Ep secretion induced by hypoxia (1% O2) without affecting cell growth. Forskolin and cholera toxin, each of which increased the levels of cyclic AMP in the Hep G2 cells by 40-fold, produced a significant (P less than 0.05) further increase in Ep secretion in the presence of hypoxia.(ABSTRACT TRUNCATED AT 250 WORDS)

Previous studies have shown that bleomycin-induced pulmonary fibrosis is accompanied by elevated levels of calcium and calmodulin, which are important in the regulation of many biologic processes. The authors have further extended these observations and assessed the effect of a calmodulin inhibitor, trifluoperazine, on bleomycin-induced lung damage with biochemical, morphometric, and bronchoalveolar lavage techniques. The cumulative mortality due to bleomycin was not significantly reduced in animals receiving trifluoperazine. Trifluoperazine had no apparent effect on lung levels of collagen and DNA elevated by bleomycin. However, morphometric studies showed that the volume density of the lesion, the volume of amorphous material and interstitial inflammation, and the number of monocytes within lesions were less in the lungs of bleomycin-treated hamsters receiving trifluoperazine daily. When compared with hamsters treated with bleomycin alone, animals treated with both bleomycin and trifluoperazine had significantly fewer lymphocytes in their bronchoalveolar lavage fluid. The data suggest that trifluoperazine reduced the acute inflammation which accompanies bleomycin pneumotoxicity but did not affect the subsequent development of pulmonary fibrosis. It has been postulated that the observed antiinflammatory action of trifluoperazine may be due to inhibition of calmodulin-dependent leukocyte functions.

To control post-operative bleeding from the resected cavity in transurethral resection of the prostate (TUR-P), 20,000 units of thrombin dissolved in 5-20 ml of physiological saline was injected into the cavity in one shot with a new type of 3-way Foley catheter. The catheter has two holes close to the balloon, and the holes provide outlets through which the solution is directly injected into the cavity. Out of 23 patients between 58 and 87 years old, 11 were given thrombin and 12 were control cases. In each case, urine blood volume (ml/hr) was determined before (a) and after (b) injection. The hemostatic effect was assessed with a "hemostatic index" of b/a. There was a statistically significant difference in the hemostatic indices between thrombin group and the control. (p less than 0.02) This new hemostatic method proved to be a very useful easy bed-side technique to control bleeding even in sudden bleeding after TUR-P. No serious side effects were found during the course of this study.

To characterize mouth to anus transit of intestinal contents, polyethylene glycol (PEG) was given as an intestinal marker to 11 healthy infants, and daily fecal collections were analyzed for PEG concentration per unit of dry stool weight for 9-15 d. Fecal PEG excretion followed first-order kinetics. Thus, half-life (t1/2) and volume of distribution (Vd) of PEG in the gut could be computed for each of seven infants who received continuous daily PEG doses and t1/2 only for four infants who received a single PEG dose. t1/2 of PEG in the gut was 0.99 +/- 0.48 d (means +/- SD). Vd for PEG in the gut was 18.74 +/- 15.38 g of fecal dry weight. We propose that whole-gut transit be expressed in terms of t1/2 and Vd of intestinal contents because these may better characterize the changes in intestinal transit that occur with disease or dietary modifications.