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Bovine aortic endothelial cells (BAEs) were used as a model system to study the nature and origin of protein(s) in the extracellular matrix that bind to type 1 plasminogen activator inhibitor (PAI-1). Matrix samples were fractionated by SDS-PAGE and analyzed by PAI-1 ligand binding and by immunoblotting using antibodies to vitronectin (Vn). PAI-1 bound primarily to two Vn-related polypeptides of Mr 63,000 and 57,000, and both of these partially degraded polypeptides were present in the culture serum. Radiolabeling experiments failed to detect significant Vn biosynthesis by BAEs (less than 0.03% of total), or by human umbilical vein endothelial cells and HT 1080 cells. The binding of PAI-1 to Vn was relatively specific since direct binding studies failed to demonstrate significant interactions between PAI-1 and other matrix proteins (e.g., fibronectin, type IV collagen, laminin, or matrigel). Kinetic studies indicate that PAI-1 rapidly accumulates in the matrix when BAEs are plated on Vn, appearing in the conditioned medium only after a significant lag period (1-2 h). However, no PAI-1 was detected in the matrix when the cells were plated on fibronectin-coated dishes, and there was no lag period for PAI-1 accumulation in the medium. These results indicate that PAI-1 binds specifically to serum-derived Vn in the matrix, and suggest that the composition of both the matrix and serum itself may influence the pericellular distribution of this important inhibitor.

We compared the effect of topical 0.5% tetracaine, 1:2,000 epinephrine, and 11.8% cocaine (TAC) with 1% lidocaine infiltration on bacterial proliferation in experimental lacerations. Forty-eight lacerations were made on the backs of Hampshire pigs, inoculated by injection with infectious doses of Staphylococcus aureus and randomly anesthetized with either topical TAC or lidocaine infiltration. Wounds were sutured, and quantitative cultures were obtained by excision after 48 hours. The mean log10 bacteria per gram of tissue for wounds anesthetized with TAC was 6.818 (95% confidence interval [CI], 6.07 to 7.54) compared with 6.820 (95% CI, 5.91 to 7.75) for those treated with lidocaine; this difference was not significant (P less than .05 by paired two-tailed t test). The probability of failing to detect an intergroup difference of 0.5 log10 bacteria per gram was less than .0001. TAC does not increase bacterial proliferation more than lidocaine infiltration in contaminated experimental porcine lacerations.

A number of experiments was carried out to determine the sequence of events leading to death following administration of the euthanizing agent T61. Simultaneous recordings of the EMG, EEG, ECG and end-tidal CO2 (dogs only) were obtained in acutely instrumented rabbits and dogs. Results show that following T61 administration the loss of consciousness and loss of muscle activity occurred simultaneously. Vocalization and increased muscle movement occurred in the initial phase of the injection in 3 of 8 dogs, injected with T61 or butyramide. From this study it was concluded that the presence of the muscle relaxant does not pose an ethical problem for the use of T61 as an euthanizing agent, but our results suggest that the use of T61 may have some emotionally unpleasant side-effects.

Epithelial-cell function requires cellular polarity in which apical membrane surfaces have unique characteristics and cellular organelles are stratified. Physiological investigations of endometrial epithelial cells would be enhanced greatly by the ability of a method to polarize cells in culture. This study investigates the effects of different substrata on polarization of cultured bovine endometrial epithelial cells. Fetal bovine endometrial epithelial-cell lines were developed from explant outgrowth. Epithelial monolayers were subcultured onto amniotic membranes, Millicell-HA membranes, or Millicell-CM membranes coated with rat-tail collagen, Matrigel, laminin, Vitrogen, or fibronectin. Cultures on these substrata were maintained at the air/liquid interface. Cells grown on either collagen-coated or uncoated Millicell membranes also were maintained submerged in medium. Excellent polarized morphology was attained in cultures grown at the air/liquid interface on amniotic membranes and rat-tail collagen-coated membranes. Lectin-binding patterns to apical membranes of polarized epithelial cell cultures paralleled patterns of binding to bovine endometrial surfaces in vivo. Cultures on rat-tail collagen were maintained for several weeks. These methods provide a valuable system for studying the endometrium in vitro.

The vascular effects of a diuretic combination (spironolactone/altizide) were studied in 5 anuric patients undergoing dialysis by measuring changes in cardiovascular reactivity to norepinephrine (NE) and angiotensin II (AII) after infusion of incremental doses of these 2 vasopressor agents. There was a marked dose-response relation between NE and AII administration and mean (NE) or diastolic (AII) blood pressure (BP). Diuretic treatment moderated the increase in mean or diastolic BP induced by NE or AII. In addition, the pressor doses of NE and AII, which are also parameters of vascular reactivity, were significantly increased after diuretic treatment (NE, +101%; AII, +163%). Right atrial and pulmonary capillary wedge pressures increased along with BP in response to NE. Diuretic treatment also moderated the increase in right atrial and pulmonary capillary wedge pressures induced by NE. These results suggest that the antihypertensive action of spironolactone and altizide in combination is mainly due to a direct effect on resistance and capacitance vessels. The mechanisms by which diuretics modify the cardioreactivity remain poorly understood; however, they may modify electrolyte transport (sodium and calcium) across vascular smooth muscle cell membranes or stimulate prostaglandin release.

To investigate the mechanisms by which acidosis depresses cardiac function, a Langendorff isolated, perfused rat heart preparation was studied using 31P-magnetic resonance spectroscopy. Isolated hearts were subjected to normal perfusion conditions or moderate or severe metabolic acidosis. Acidosis resulted in dose-dependent decreases in O2 consumption (VO2), rate of developed ventricular pressure (dP/dt), intracellular pH (pHi), and the tissue concentration of creatine phosphate (PCr) after 30 min. Acidosis did not affect ATP concentration but resulted in significant increases in the concentrations of total and monovalent inorganic phosphate (Pi and Pim). When metabolic determinants of the change in dP/dt with acidosis were examined, changes in pHi, extracellular pH, and tissue concentrations of Pi and Pim all correlated significantly with the change in dP/dt. In a second set of experiments using the severe acidosis model, normal saline, 1 M sodium bicarbonate, and the experimental buffer Carbicarb (1 ml/kg body wt) were infused in a crossover manner at 20-min intervals. None had significant effects on tissue concentrations of ATP, PCr, Pi, or PCr/Pi. Administration of bicarbonate resulted in an increase in VO2 without significant effects on pHi or dP/dt. In contrast, Carbicarb effected little change in VO2 but resulted in sustained increases in pHi and decreases in Pim and marked increases in dP/dt. These data suggest that the decrease in cardiac function that occurs during acidosis may be related to the metabolic consequences of reductions in pHi and that a buffer that causes intracellular alkalinization may cause functional improvement in this model.

The effect of brain temperature and anesthesia on ischemic neuronal damage was studied in the hippocampal formation using the four vessel occlusion model in awake and anesthetized rats. Neuronal damage was assessed by immunocytochemistry and silver impregnation of tissue sections. The degree of ischemia was monitored by recording spontaneous and evoked electrical activity from the hippocampus and dentate gyrus in all animals. In addition, the hippocampal temperature and oxygen tension were also recorded using a chamber-type thin-film microelectrode in the anesthetized animals. Fifteen minutes ischemia in the awake animals caused greater neuronal damage and mortality of animals than 30 min ischemia in anesthetized rats. The temperature of the brain was found to drop by 4-6 degrees C during complete forebrain ischemia in the latter group. Neuronal damage was observed infrequently in the hippocampus of these animals. When the brain temperature was kept constant at the preischemic level during 30 min occlusion, all animals died within a day, while after 15 min occlusion the majority showed an almost complete degeneration of CA1 pyramidal cells and hilar somatostatin immunoreactive neurons. Following 15 min ischemia, the awake animals showed a similar cell loss in the CA1 region and the hilus. It is concluded that, in the anesthetized animals prepared for acute recording, the decreased temperature of the brain during ischemia is a major factor in protecting neurons from damage, but that Equithesin anesthesia also has a significant protective effect. Consistent ischemic degeneration occurs in awake animals by four vessel occlusion, if the brain temperature is controlled and the completeness of ischemia is monitored by recording spontaneous and evoked electrical activity with chronic electrodes.

The effects of anabolic implants, growth phase (growing vs finishing) and rate of growth on the priorities for protein and fat deposition were determined in yearling cattle. Santa Gertrudis crossbred yearling steers weighing 290 kg were individually fed diets varying in forage and grain content and either not implanted (n = 16) or implanted (90-d intervals) with Ralgro (n = 13) or Synovex-S (n = 12) implants. The cattle were fed toward a similar expected final empty BW (455 kg). Initial and interim empty body composition was measured via deuterium oxide dilution; final composition was determined by carcass specific gravity. During a 100-d growing phase, rates of protein gain were increased (P less than .12) to 118 and 131 g/d from 98 g/d for Ralgro and Synovex vs nonimplanted cattle, respectively. Concurrently, the fraction of protein in empty body growth was increased (P less than .09) from 17.5% for controls to 23.8 and 19.7% for Ralgro- and Synovex-implanted steers, respectively. This change in protein growth occurred concomitant with mobilization of fat and a reduction (P less than .04) in fat gain with Ralgro and Synovex implants. During the 136-d finishing phase, protein accretion was 115 and 132 vs 93 g/d for Ralgro- and Synovex-implanted cattle vs nonimplanted cattle; this represented a 24 and 42% increase (P less than .03) with Ralgro and Synovex, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

The authors report the advantages of a new anticoagulant-antiaggregant mixture that avoids the deleterious effects of ethylenediaminotetraacetic acid (EDTA) on mean platelet volume and also prevents EDTA-induced platelet clumping. It is suitable for routine cell counting and sizing with the Coulter Counter S-Plus STKR. The values of the common hematologic parameters agree well with those from EDTA-treated samples and are stable for at least eight hours after sampling.

The role of Na+ and Na(+)-H+ exchange in the stimulation of ornithine decarboxylase (ODC) activity has been investigated in a human colon adenocarcinoma cell line, LoVo. Asparagine (Asn; 10 mM) or 10% fetal bovine serum (FBS) increased ODC activity from undetectable levels to greater than 500 pmol CO2.mg protein-1.h-1 in 4 h. This increase could be reduced 50% by concentrations of Na(+)-H+ exchange inhibitors that did not reduce protein synthesis. (approximately 0.2 mM for amiloride and 0.05 mM for hexamethyleneamiloride). Asn was able to double the uptake of 22Na+, whether an ionic (choline chloride) or nonionic (D-mannitol) substance was substituted for Na+, and the substitution of these compounds as well as N-methyl-glucamine for Na+ largely prevented the stimulation of ODC by Asn. Another factor influencing ODC activity was extracellular pH (pHo). When pHo was lowered, intracellular pH (pHi) also fell, and ODC activity was reduced. When pHo was raised, pHi also rose, and ODC activity increased. The well-known correlation between increased pHi and Na+ uptake with the stimulation of growth may be due to their influence on ODC activity.

The purpose of this study was to evaluate the histologic changes that occur in an area of rat skeletal muscle injected with a local anesthetic and steroid combination. Thirty adult male laboratory rats were assigned to one of six groups. Group 1 received bupivicaine injected into the right gastrocnemius muscle; Group 2 received procaine; Group 3 received bupivicaine and triamcinolone; Group 4 received procaine and triamcinolone; Group 5 received triamcinolone; and Group 6 received normal saline. Animals were sacrificed at 24 hours and at 1, 2, 3, and 4 weeks. The saline, steroid, and procaine groups showed minimal tissue reaction. The procaine/steroid group had focal areas of inflammation at 24 hours but none on subsequent evaluations. The bupivicaine group showed moderate localized necrosis of muscle fibers and a mild inflammatory cell response at 24 hours. Regeneration of muscle fibers was seen at 1 week, and at 3 weeks the tissue had returned to normal. The bupivicaine/steroid group showed extensive localized necrosis of muscle fibers with a heavy inflammatory cell response at 24 hours. These areas of necrosis persisted throughout the 4-week post-injection period, although some muscle fiber regeneration and fibrosis were also noted. This study shows that bupivicaine produces more tissue reaction than procaine and that the addition of steroid to bupivicaine increases the initial tissue damage and prolongs the healing phase.

It is frequently assumed that, in general, combinations of antibiotics and similar chemotherapeutic agents are more efficient than the single chemotherapeutic, and that they can be combined without disadvantages for the activity of the single component. However, the specific properties of the combined substances, such as their antimicrobial spectrum and resistance, their pharmacokinetics and side effects, as well as their physical properties and formulation, have an enormous impact on the activity of combined chemotherapeutics. They may lead to an advantageous, but also to an indifferent or diminished activity of the combination in patients. Some of the important requirements for an advantageous combination of antibiotics and similarly functioning compounds will be reviewed and ways of avoiding mistakes are described.

The consequences of using surgical bone wax are not well studied. We evaluated the infection-promoting potential of sterile bone wax in a rat model of chronic Staphylococcus aureus osteomyelitis. The addition of bone wax greatly reduced the quantitative bacterial inoculum (log colony-forming units) required to establish chronic osteomyelitis in 50% and 100% of challenged animals. The 50% infection rate was reduced from log 6.9 to 2.6 and the 100% infection rate from 8.2 to 4.4, respectively (p less than 0.015, t test for parallelism). Separate experiments were done 10 to 30 minutes after inoculation with only log 6.4 staphylococci. Tibiae of animals that received bone wax yielded more organisms than those that did not (log 2.76 +/- 0.68 versus 1.72 +/- 0.94, p less than 0.01). At 24 hours quantitative colony counts were not significantly different whether animals received wax or not (log 5.02 +/- 0.42 versus 4.43 +/- 0.65, p greater than 0.09). These studies suggest that the routine surgical use of bone wax should be reassessed.

In an effort to develop effective combination treatments for use with radiation against solid tumors, the cytotoxic effects of the addition of mitomycin C or porfiromycin on treatment with Fluosol-DA/carbogen (95% O2/5% CO2) breathing and radiation in the FSaIIC tumor system were studied. In vitro mitomycin C and porfiromycin were both preferentially cytotoxic toward hypoxic FSaIIC cells. After in vivo exposure, however, the cytotoxicity of mitomycin C toward single cell tumor suspensions obtained from whole tumors was exponential over the dose range studied, but for porfiromycin a plateau in cell killing was observed. With Fluosol-DA/carbogen breathing and single dose radiation, addition of either mitomycin C or porfiromycin increased the tumor cell kill achieved at 5 Gy by approximately 1.2 and 1.0 logs, respectively. Less effect was seen with addition of the drugs at the 10 and 15 Gy radiation doses. In tumor growth delay experiments, the addition of either mitomycin C or porfiromycin to Fluosol-DA/carbogen breathing and radiation resulted in primarily an additive increase in tumor growth delay. The survival of Hoechst 33342 dye-selected tumor cell subpopulations indicated that Fluosol-DA/carbogen breathing increased the cytotoxicity of radiation (10 Gy) more in the bright cell subpopulation (4-fold) than in the dim cell subpopulation (2-fold) resulting in an overall 4-fold sparing of the dim subpopulation. Mitomycin C and porfiromycin were both more toxic toward the dim cell subpopulations. Addition of mitomycin C or porfiromycin to Fluosol-DA/carbogen breathing and radiation (10 Gy) resulted in a primarily additive effect of the drugs and radiation killing in both tumor cell subpopulations. Thus, with mitomycin C/Fluosol-DA/carbogen and radiation there was a 2-fold sparing of dim cells and with porfiromycin in the combined treatment a 1.6-fold sparing of the dim cell population. Our results indicate that treatment strategies directed against both oxic and hypoxic tumor subpopulations can markedly increase the tumor cell kill achieved by radiation.

Ninety crossbred beef heifers averaging 260 kg were blocked by weight and allotted randomly to 15 pens of six heifers each, with three replicates per treatment. The treatments were spayed heifers (S); intact heifers (I); S + Synovex-H7 (SH); I + Synovex-H (IH); and S + Synovex-S7 (SS). The heifers were spayed vaginally with a Willis instrument; intact heifers were rectally palpated for reproductive soundness. Heifers were fed a growing diet (55 d), re-implanted and subsequently fed a finishing ration (73 or 101 d). All heifers were observed for estrus daily at approximately 0600 and 2000 until d 69. A jugular vein blood sample was obtained from each heifer on d 48, 55, 62 and 69 for blood progesterone analysis. Results of blood progesterone analysis and reproductive tract examination indicated that two spayed heifers were incompletely spayed, a 96% success rate. The SH and SS heifers continued to exhibit estrous behavior despite being successfully spayed. Heifers implanted with Synovex-H had greater (P less than .01) ADG and were more efficient (P less than .03) than nonimplanted heifers. The SS heifers had greater ADG (P less than .05) during the finishing phase than SH heifers, but no advantage of SS over SH was apparent during the growing phase. The overall ADG response to implantation was fourfold greater (P less than .07) in the spayed heifers than in the intact heifers (32 vs 8%). Heifers implanted with Synovex-H had greater adjusted hot carcass weight (P less than .02) and ribeye area (P less than .002) than nonimplanted heifers. For adjusted live weight and ribeye area, the response to implantation was approximately threefold greater in the spayed heifer than the intact heifer. These results demonstrate that spaying and implanting heifers can increase rates and efficiency of gain even though behavioral estrus is not eliminated.

A motorized and computer-interfaced phoropter was used to track the development of cycloplegia and recovery of accommodation over a 60-min period, after the topical application of a phenylephrine 5%-tropicamide 0.8% drug combination (Phenyltrope). Phenyltrope was introduced into the Canadian market about 2 years ago (Compendium of Pharmaceuticals and Specialties, 1987), and advertised as a fast acting cycloplegic and mydriatic drug. Here we report the results of our investigation of the depth of action and the temporal aspects of cycloplegia for this drug combination as a function of iris color. We also compare the action spectrum of Phenyltrope to that of tropicamide 1% under similar test conditions. Our results indicate that the latency for cycloplegia was shorter for tropicamide, the maximum rate of accommodative loss similar for both drugs, and the resultant cycloplegia at 20 min deeper for Phenyltrope. Recovery from induced cycloplegia was greater for tropicamide 60 min after drug administration. For both Phenyltrope and tropicamide, no significant differences in any of the parameters investigated were observed as a function of iris color. We conclude that even though Phenyltrope induced a measurably deeper cycloplegia than did tropicamide, the amount of residual accommodation present at 20 min (about 38%) is insufficient for most refractive purposes.

In order to demonstrate an immunomodulating effect of cotrimoxazole, we investigated its influence on some macrophage (M phi) functions in culture: P815 tumor cells killing, Toxoplasma gondii killing, production of free oxygen radicals by luminol-dependent chemiluminescence, prostaglandins and leukotrienes secretion evaluated after incorporation of tritiated arachidonic acid. In vitro, cotrimoxazole inhibited in a dose-dependent fashion the chemiluminescence of murine resident peritoneal or guinea pig alveolar M phi. Production of prostaglandin (PG) 6-keto-F1 alpha, PGF2 alpha, and 5-hydroxyeicosatetraenoic acid by resident peritoneal M phi was also inhibited. However, PGD2 synthesis by alveolar M phi was enhanced. A second study was performed on peritoneal M phi, resident or elicited in vivo by one intra-peritoneal injection of an extract from Mycobacterium Tuberculosis membranes and obtained from mice pretreated or not by cotrimoxazole per os. Resident M phi from cotrimoxale-treated animals showed increased production of leucotriene B4 compared to M phi from controls. 6-keto-PGF1 alpha and free oxygen radicals production by elicited M phi was greatly enhanced by cotrimoxazole whereas thromboxane B2 was reduced. Finally cotrimoxazole enhanced intracellular killing of Toxoplasma gondii and cytotoxicity for tumor cells P815 by resident but not by elicited M phi. It is concluded that cotrimoxazole can modulate MO activation and some M phi functions involved in immune homeostasis. This data could help to understand why an antibiotic such as cotrimoxazole, which is known to be frequently used in immunocompromised hosts, is also efficient in Wegener's granulomatosis.

The analgesic effect of topical application of local anaesthetics on the gingival mucosa and the absorption of the local anaesthetics into the blood were investigated in healthy volunteers by using a 5% eutectic mixture of the local anaesthetics lignocaine (100 mg) and prilocaine (100 mg) plus emulsifier (EMLA) or 10% lignocaine (200 mg) spray (Xylocain). The pain threshold on labial gingiva was measured by using the stimulator of an EMG-apparatus and a pair of stimulating electrodes, specially constructed for this purpose. There were no differences between the two methods in producing analgesia which was at its maximum in 13 to 14 minutes, on average. Sensation of the gingiva (pain thresholds), had usually returned to normal within 30 minutes. In the dosages used, the absorption of the local anaesthetics was more rapid after the mixture application than after spray application. No toxic reactions occurred.

A longitudinal controlled study on the effect of Diane and Microgynon 50 (Micro 50) contraceptive pills on the haemostatic mechanisms in 59 Chinese subjects was carried out. Diane, containing the potent anti-androgen progestagen, cyproterone acetate was used as a contraceptive and in subjects requiring anti-androgenic treatment and fertility control. Enhanced fibrinolytic activity with raised plasminogen and alpha 1-antitrypsin (a1-AT) was observed in all subjects after pill treatment. The changes observed after Diane treatment suggests a predispostion to hypercoagulation as evidenced by increases in Fibrinogen, Factors VII and X with accelerated Prothrombin Time (PT), Activated Partial Thromboplastin Time (APTT) and Reaction and Clotting Time (r + k). Maximum elasticity of the clot was raised after Diane 50 treatment when compared with pre-treatment levels. More changes in haemostasis were seen after treatment with Diane containing 35 ug and 50 ug ethinyl estradiol (EE) formulation than during Micro 50 treatment. The results from this study show that Diane even with 35 ug EE formulation caused coagulation changes which were more pronounced than the 50 ug EE Microgynon. The finding is at variance with others that ascribe haemostatic changes observed to increased oestrogen content in a given pill formulation and so merits confirmation in a larger study.

We have observed the influence of Radix Salviae Miltiorrhizae and Ligustrazini Hydrochloridum on the elastase activity and the protective effects of the two medicines on the elastic fibers. The results showed that Radix Salviae Miltiorrhizae and Ligustrazini Hydrochloridum can inhibit the activity of porcine pancreatic elastase and human sputum elastase. These two drugs can protect arterial and pulmonary elastic fibers from the destruction of elastase. Since these traditional Chinese herbs have little toxic effect and no antigenic character, they can be used in the preventing emphysema. The study suggest that it is useful Radix Salviae Miltiorrhizae and Ligustrazini Hydrochloridum are as favourable elastase inhibiting agents, which may be used in the prevention of emphysema extensively.

Three experiments were carried out to find the optimal concentration of DMSO and glycerol in BF5 diluent for freezing rabbit spermatozoa. Semen was diluted 1:1 with diluent A (BF5 + DMSO) at 25 degrees C and diluted further 1:1 with diluent B (diluent A + glycerol) after cooling down to 5 degrees C. Diluted semen was frozen immediately and stored in liquid nitrogen. Maximum percentages for motility and normal acrosomes were obtained in the presence of 12% DMSO (as expressed in diluent A) and 3% glycerol (final concentration) after thawing.

The antibacterial activity of BIPP and its constituents against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa was measured by growth inhibition tests. BIPP was found to have negligible antibacterial activity. In addition, no release of iodine from BIPP was detected over a 4-week period. It is proposed that much of the evident antibacterial activity of BIPP may be a reflection of the meticulous surgical debridement that accompanies its use. In addition, BIPP makes the impregnated gauze impervious to blood and body fluids ensuring little nutrition for bacteria to thrive in its interstices.

An assessment was made of the effects of topical application of a eutectic mixture of local anaesthetics (EMLA cream) in a number of species of laboratory animals. Application of EMLA cream enabled percutaneous insertion of catheters into the cephalic vein in dogs and cats and the marginal ear vein in rabbits without causing any detectable pain or discomfort. Application to the tail in rats prior to percutaneous cannulation of the lateral tail vein did not produce a significant reduction in the behavioural responses to venepuncture. EMLA cream represents a useful refinement of current techniques for intravenous injection in some species, and is especially valuable when the procedure is to be undertaken by an inexperienced operator.

The effect of polyphytochol representing a combination of vegetable agents was studied on models of acute toxic hepatitis caused by CCl4 and cholecystitis. The efficiency of the drug in pathology of the hepatobiliary system was shown: it decreases cholestasis acting as a choleretic and bile-secreting drug, acts on the main pathogenetic links in hepatitis, exerts a marked membrane-stabilizing effect, influences favourably the processes of synthesis in the liver.

The pharmacokinetics of FCE 22101 were studied in eight healthy male subjects who received FCE 22101 intravenously alone or together with imipenem/cilastatin which was given to inhibit dehydropeptidase-I, a renal enzyme metabolizing penem and carbapenem antibiotics. The kinetics of FCE 22101 were also studied following oral administration of its acetoxymethyl ester, FCE 22891. For comparative purposes, the kinetics of imipenem and cilastatin, given alone or together with FCE 22101, were calculated. Intravenously administered FCE 22101 at a dose of 250 mg gave peak plasma concentrations of about 12 mg/l and the plasma half-life was about 60 min. Co-administration of FCE 22101 with imipenem/cilastatin did not affect the plasma kinetics of FCE 22101, nor did FCE 22101 influence the kinetics of imipenem or cilastatin. Cilastatin increased the urinary recovery of FCE 22101 from 17.5% to 53.0% with FCE 22101 alone to 73.2% to 91.8% when it was given with cilastatin. There was a high correlation between the urinary recovery of FCE 22101 in this study and that of imipenem given alone to the same subjects in previous studies; subjects who were high metabolizers of imipenem were also high metabolizers of FCE 22101. When FCE 22891 was given orally at a dose of 500 mg (corresponding to 400 mg of FCE 22101 free acid), peak concentrations of 2.2 to 6.1 mg/l were found. The absorption was rapid with peak concentrations achieved 20 to 80 min after administration. In comparison with imipenem, FCE 22101 seems to undergo less non-renal metabolism.

The hemorheologic alterations were studied in experimental acute hemorrhagic necrotizing pancreatitis (AHNP) which was induced by injection of 5% sodium taurocholate into the main pancreatic duct. The results showed that during the early stage of AHNP, all of hemorheologic parameters were significantly elevated, and continually increased at the rest intervals. The authors suggested that the blood viscosity and hematocrit increase, red blood cells tend to aggregate, and erythrocyte deformation decrease are the causation of pancreatic microcirculatory disturbances and promoting pancreatic progressive necrosis. After using low molecular dextran and Salvia miltiorrhizae, the hemorheologic alterations in AHNP were recovered, and the histologic observation improved. Our data indicate the hemorheologic disturbances is one of the key factors in the pathogenesis of AHNP.

The antihypertensive effect of a fixed dosage combination of the cardioselective beta-adrenoceptor blocker, atenolol, and the oral thiazide-like diuretic, chlorthalidone (Tenoretic) was studied in 24 hypertensive Nigerians in a double-blind, cross-over comparison with three other treatments. These were atenolol alone, 100 mg daily, chlorthalidone alone, 25 mg daily, and atenolol (100 mg) plus chlorthalidone (25 mg) daily taken as separate formulations. Tenoretic was taken as a once-daily tablet containing 100 mg atenolol plus 25 mg chlorthalidone. The order of administration of the drugs was randomized. Each drug was taken for 4 weeks. The results showed that atenolol and chlorthalidone lowered blood pressure to the same extent. Combination of the two drugs whether taken separately or in fixed-dosage combination was better than either product singly. The drugs were well tolerated.

Antacids containing Al(OH)3 have been shown to stimulate the protective and reparative processes in the gastric mucosa exposed to various ulcerogens but the mechanisms of these effects are unknown. This study was designed to determine the role of intragastric pH and mucosal formation of prostaglandins (PG) in antacid-induced protection of gastric mucosa against ethanol or aspirin damage. Maalox 70 and its active component, Al(OH)3, were used alone at the original pH (7.5 and 6.0, respectively) and lower pH or were combined with ranitidine or histamine before intragastric administration of 100% ethanol or acidified aspirin to induce acute mucosal lesions. Maalox and Al(OH)3 were relatively more effective to protect against the damage due to ethanol, and to some extent also due to aspirin, when given at a pH (pH 2) lower than at their original pH. Pretreatment with ranitidine, which itself did not change ethanol-induced damage, greatly reduced the protection afforded by Maalox or Al(OH)3, whereas pretreatment with histamine enhanced this protection. Maalox and Al(OH)3 at their original or low pH did not alter significantly the capability of gastric mucosa to generate PG but resulted in a significant increase in luminal release of PG. Pretreatment with indomethacin to reduce mucosal generation of PG failed to affect the protective effects of Maalox or Al(OH)3 at their original or lower pH. We conclude that Maalox 70 and Al(OH)3 protect the gastric mucosa against ethanol damage but that this protection requires the presence of acid and may not depend entirely upon the mucosal production of PG.

The biocompatibility was studied of 3 hemostatic agents (Astroplast, Hemastogen and bone wax) implanted into experimental bone cavities together with their clearance from the implantation sites. The 3 agents had an acceptable hemostatic effect during the surgical procedure. However, marked differences regarding tissue reactions in the experimental bone cavities, as well as at the internal bone surfaces, were found between them. Astroplast and Hemastogen appear to provoke more adverse tissue reactions, including foreign body giant cell formation, compared with bone wax. All 3 hemostatic implants showed markedly delayed bone healing. It was concluded that their biocompatibility was not ideal but acceptable in view of the clinical advantages.

Rings of rat aorta cultured in Matrigel, a reconstituted gel composed of basement membrane molecules, gave rise to three-dimensional networks composed of solid cellular cords and occasional microvessels with slitlike lumina. Immunohistochemical and ultrastructural studies showed that the solid cords were composed of endothelial sprouts surrounded by nonendothelial mesenchymal cells. The angiogenic response of the aortic rings in Matrigel was compared to that obtained in interstitial collagen, fibrin, or plasma clot. Morphometric analysis demonstrated that the mean luminal area of the microvascular sprouts and channels was significantly smaller in Matrigel than in collagen, fibrin, or plasma clot. The percentage of patent microvessels in Matrigel was also markedly reduced. Autoradiographic studies of 3H-thymidine-labeled cultures showed reduced DNA synthesis by developing microvessels in Matrigel. The overall number of solid endothelial cords and microvessels was lower in Matrigel than in fibrin or plasma clot. A mixed cell population isolated from Matrigel cultures formed a monolayer in collagen or fibrin-coated dishes but rapidly reorganized into a polygonal network when plated on Matrigel. The observation that gels composed of basement membrane molecules modulate the canalization, proliferation, and organization into networks of vasoformative endothelial cells in three-dimensional cultures supports the hypothesis that the basement membrane is a potent regulator of microvascular growth and morphogenesis.

Extracellular microelectrode method and stereotaxic technique of brain were used to investigate the effects of Radix Salviae Miltiorrhizae on the discharges of visceral pain in the posterior nucleus of thalamus(po)in cats. Experiments showed that Radix Salvia Miltiorrhizae inhibited the discharges of visceral pain in po, Radix Salviae Miltiorrhizae acts directly on the peripheral nerve trunk and cannot block its exciting conduction. This suggests that the analgesic effect of Radix Salviae Miltiorrhizae is exerted via central nervous system.

Myocardial salvage after reperfusion may be limited by neutrophil-mediated microvascular damage. The effect of the perfluorochemical, Fluosol-DA, and its various components on neutrophil adherence, cytotoxicity, and proteolytic enzyme release was examined on sheep large and small vessel endothelial cells in vitro. Cells were studied under normoxic (N) and anoxic conditions (A). Various concentrations of Fluosol (10%, 25%, and 50%) significantly reduced neutrophil adherence under both experimental conditions [mean 22 +/- 3.25% versus 7 +/- 0.8% (N) and 20 +/- 3.2% versus 7.5 +/- 0.9% (A); P less than 0.01]. The perfluorocarbons, perfluorodecalin (PFD), and perfluoro-tripropylamine (PFTP) in a 50 volume/percent concentration exhibited profound effects on adherence, particularly on cells subjected to anoxia (51% and 69% reduction in adherence, respectively; P less than 0.01). No effect on adherence was observed with other components, including the detergent, pluronic F68. A 25% reduction (P less than 0.02) in endothelial cytotoxicity was noted when neutrophils were preincubated with Fluosol. However, pretreatment of endothelial cells with Fluosol did not inhibit neutrophil adherence. Neutrophils stimulated with cytochalasin B and FMLP showed a significant reduction in lysozyme release after incubation with Fluosol (28 +/- 5% versus 17 +/- 4%; P less than 0.01). This study demonstrates that Fluosol significantly attenuates neutrophil adherence, cytotoxicity, and enzyme release in an in vitro model of microvascular injury. It also suggests that prevention of neutrophil-mediated microvascular damage may be an important mechanism whereby Fluosol enhances myocardial salvage after ischemia and reperfusion.

The role of splenocytes and erythrocytes in showing an extracorporal action by terrilytin and lysozyme was studied. The extracorporal effect of terrilytin was to a greater extent mediated by the spleen cells adhering to the plastic while the extracorporal effect of lysozyme was mainly mediated by the heavy ("old") erythrocytes. The heat treatment at a temperature of 42 degrees C for 15 minutes did not abolish the terrilytin extracorporal effect mediated by the erythrocytes but completely abolish the similar effect induced by lysozyme which bound to the erythrocyte membrane. After exposure of the erythrocytes to terrilytin, the strength of the lysozyme binding increased and there was a respective increase in the immunostimulating activity of the erythrocytes.

Recent studies have demonstrated an increase in carbon dioxide (CO2) tension (PCO2) in both mixed venous and coronary vein blood early in the course of cardiac arrest and cardiopulmonary resuscitation. Because increased PCO2 in the myocardium correlates with both ischemic injury and depression of contractile function, the effects of hypertonic solutions of either the CO2-"generating" sodium bicarbonate (NaHCO3) buffer, a mixture of sodium carbonate (Na2CO3) and sodium bicarbonate (carbicarb) acting as a CO2-"consuming" buffer, or saline placebo (NaCl) were compared during cardiopulmonary resuscitation in 25 healthy minipigs. Both buffer agents significantly increased the pH and HCO3- of arterial, mixed venous and coronary vein blood. Bicarbonate increased whereas carbicarb reduced blood PCO2 in the systemic circuit as anticipated. However, neither the PCO2 nor the lactate content of coronary vein blood was favorably altered by buffer therapy. Four of eight animals treated with bicarbonate, five of eight treated with carbicarb and six of nine placebo-treated animals were successfully resuscitated and had a comparable 24 h survival rate. Coronary perfusion pressure during precordial compression, a critical determinant of resuscitability, was transiently decreased by each of the hypertonic solutions. Accordingly, neither CO2-generating nor CO2-consuming buffers mitigated increases in coronary vein PCO2 or improved the outcome of cardiopulmonary resuscitation under these experimental conditions.

We have determined the depth and duration of analgesia to needle insertion after topical application of EMLA cream (Eutectic Mixture of Local Analgesics). EMLA was applied for 30, 60, 90 and 120 min and the sensory and pain threshold depths were determined before analgesia (1.0 and 1.9 mm, respectively) and up to 4 h after the cream was removed from the skin. The maximal depth of analgesia (approx. 5 mm) was observed 30 min after a 90-min application and during the 60-min period after a 120-min application of EMLA cream, for both sensory and pain thresholds. For application times shorter than 120 min, the depth of analgesia increased during the period after removal of the cream. This suggests new guidelines for the use of this topical analgesic.

This investigation evaluated the potential effect of gadolinium (Gd)-DTPA-dimeglumine on synovial membrane and joint cartilage, using macroscopic, microscopic, and x-ray fluorescent spectroscopic techniques. Thirteen New Zealand white rabbits (26 knees) were used in this study, ten receiving 500 micromolar injections of Gd-DTPA-dimeglumine in their right knees; the remainder of the knees served as controls. One injected knee had minimal joint effusion and one had mild hyperemia. Microscopically four knees exhibited mild focal hyperplasia of the synovium, another three minimal focal mononuclear cell infiltration. X-ray fluorescent spectroscopy demonstrated no evidence of Gd-DTPA-dimeglumine in the synovium or articular cartilage. Neither macroscopic nor microscopic evaluation detected any Gd-DTPA-dimeglumine related effects. Gd-DTPA-dimeglumine was found to be safe for intra-articular injection in this animal model.

Contrast media occasionally produce bradyarrhythmias defined as a 25% decrease in heart rate and/or developing atrioventricular block during coronary angiography. Twelve left coronary angiographies and seven right coronary angiographies were performed with 10 ml of diatrizoate meglumine and diatrizoate sodium (Renografin-76 [R76] or Hypaque [H76]) or iopamidol (Isovue 370 [ISO]) in a blinded randomized fashion. Heart rate decreased significantly from 135 +/- 5 to 120 +/- 5 beats/min (p less than 0.001) with R76, to 127 +/- 7 beats/min (p less than 0.01) with H76, and to 130 +/- 6 beats/min (p less than 0.05) with ISO in left coronary angiographies; more profound decrease was observed in right coronary angiographies from 134 +/- 4 to 87 +/- 18 beats/min (p less than 0.001) with R76, to 99 +/- 14 beats/min (p less than 0.001) with H76, and to 125 +/- 7 beats/min (p less than 0.01) with ISO. In 12 left coronary angiographies bradyarrhythmia was observed in five cases with R76, two with H76, and none with ISO. In seven right coronary angiographies it occurred in six with R76, three with H76, and none with ISO. The differences in the incidence of bradyarrhythmia between R76 and ISO were significant during left and right coronary angiographies (p less than 0.05 and p less than 0.01, respectively). Thus ionic contrast media produced more marked bradyarrhythmias than nonionic contrast media in coronary angiography, especially R76. Right coronary angiography resulted in more profound bradyarrhythmias than left coronary angiographies. This study suggested that nonionic contrast media (ISO) might be preferable to ionic contrast media (R76 or H76) for coronary arteriography.

In the present study we observed the absolute number of NK cells and the NK activity in 10 patients with cervical cancer. NK activity was determined before and after stimulation with Active Lipids. We reported a significant decrease of basal NK activity, but after AL stimulation NK activity was increased with statistical significance. Therefore our findings show that the reduction of NK activity in neoplastic patients is not an irreversible phenomenon since it can be restored by "in vitro" AL administration. This is important for the adoptive immunotherapy by which we could give LAK cells to the patients.

We have measured plasma concentrations of local anaesthetics, and the substance fraction of methaemoglobin (MetHb), in infants less than 3 months of age, after application of a lignocaine-prilocaine cream (EMLA). A total of EMLA 2 g was applied over four different skin areas, totalling 16 cm2, for 4 h before anaesthesia for a minor surgical procedure. Sampling was carried out before and 4, 8 and 12 h after application. Maximum MetHb values (median = 2.24%) were obtained usually at 8 h and were significantly (P less than 0.001) higher than before application (median 1.32%). The plasma concentrations of local anaesthetics were low (maximum values: prilocaine 78 ng ml-1, lignocaine 412 ng ml-1). The activity of erythrocyte MetHb reductase (cytochrome b5 reductase) was analysed. Data from a previously studied group of infants aged 3-12 months were included also. Enzyme activity did not reach adult levels until after the age of 3 months. It showed a good inverse correlation with the maximum MetHb values after application of EMLA. Although the MetHb concentrations in the infants younger than 3 months were small, the enzyme capacity may be overloaded when EMLA is administered at the same time as other MetHb-inducing agents. It is concluded that the use of EMLA should be restricted in this age group.

Type IV and interstitial collagenolytic activities were compared in human malignant and normal trophoblast cells cultured on plastic, in presence or absence of laminin in solution, on matrigel (a gel of basement membrane components) and on type I collagen gel. Laminin highly stimulated the type IV collagenolytic activity but not the interstitial collagenolytic activity, in malignant trophoblast cells. This glycoprotein had no effect on the interstitial collagenolytic activity and doubled the type IV collagenolytic activity in normal trophoblast cells. Thus malignant trophoblast cells produce preferentially the enzyme able to degrade basement membrane when in contact with laminin, and the enzyme able to degrade interstitial collagen fibers when cultured on type I collagen. On the contrary, type I collagen gel and matrigel equally increased both type IV and interstitial collagenolytic activities by normal trophoblast cells. Interactions of tumor trophoblast or normal trophoblast cells with the extracellular matrix result thus in distinct stimulations of collagenolytic activities. Increased production of type IV collagenolytic activity upon exposure to laminin appears to be specific of the metastatic phenotype.

In vitro EDTA-induced platelet aggregation is a fairly rare event but can have serious clinical consequences producing pseudothrombocytopenia and pseudoleukocytosis. Sixteen specimens with previously recognized EDTA-induced platelet aggregation were collected in a new anticoagulant-antiaggregant mixture containing trisodium citrate 17 mmol/l, pyridoxal 5'-phosphate 11.3 mmol/l and Tris 24.76 mmol/l (CPT mixture) and analyzed at various times after venepuncture with four hematological instruments: Coulter Counter S-Plus STKR, Technicon H6000, Technicon H1 and Ortho ELT-8. In CPT-anticoagulated specimens the signals and instrumental flags of platelet clumping were absent, and the platelet number correlated very well with a microscopic count from a finger stick drawn into Unopette. The complete blood count was very similar in "normal" hematological specimens either collected in K3. EDTA or in CPT, although Technicon H1 and Ortho3 ELT-8 required a suitable calibration for MCV and hematocrit in the latter mixture. Mean platelet volume was stable for up to 24 h only in CPT-collected specimens, if it was measured on a Coulter Counter S-Plus STKR. In routine hematological practice CPT can be an alternative anticoagulant to K3. EDTA, most suitable for automated complete blood count and useful in avoiding EDTA-induced platelet clumping.

The effects of single doses of a combination of 20 mg frusemide and 2.5 mg amiloride, of 20 mg frusemide and of placebo on plasma and urinary electrolytes were assessed in healthy volunteers using a randomized, crossover study. The combination significantly (P less than 0.001) increased the urinary volume compared with frusemide alone or placebo. Mean total urinary sodium was also significantly (P less than 0.001) increased by frusemide/amiloride compared with both frusemide and placebo. Mean total potassium excretion was reduced significantly (P less than 0.01) by frusemide/amiloride but total magnesium excretion was not significantly affected. The effects of frusemide/amiloride on plasma sodium, magnesium and potassium were similar to those of placebo. It is concluded that frusemide/amiloride has a greater diuretic and natriuretic effect than frusemide alone and does not increase potassium loss.

Myocardial 45Ca sequestration was studied in dogs during 60 minutes of global ischemia and 30 minutes of reperfusion using cardiopulmonary bypass (CPB) and myocardial function was studied before and after CPB. Group A (n = 5), as a control, received cold hyperkalemic cardioplegic solution, Group B (n = 5) received the same solution plus verapamil (150 micrograms/kg/L) and Group C plus specific activity (TSA = DPM x 10(4)/g) and plasma specific activity (PSA = DPM x 10(4)/ml) were determined by biopsy before and after release of the cross-clamp The results showed that myocardial function in Group B and C were better than that in Group A (P less than 0.01). It is suggested that verapamil and Salvia miltiorrhiza Bunge effectively control myocardial calcium sequestration during early reperfusion and reduce myocardial reperfusion injury As to myocardial protection, cardioplegia with verapamil and Salvia miltiorrhiza Bunge were superior to hyperkalemic alone. Verapamil cardioplegia was still better than Salvia miltiorrhiza Bunge.

The ascorbic acid pretreated beagle dog was used to demonstrate the phenomenon of ion-trapping and the influence of three different antacid treatments on the serum and gastric fluid concentration of meperidine: pretreatment with magnesium-aluminum hydroxide (MA), pretreatment with cimetidine (C), and periodic total aspiration of gastric fluid (ASP). A significant correlation between pH and the meperidine concentration in gastric fluid was found; the lower the pH the higher is the secretion of meperidine into gastric fluid. The gastric fluid meperidine concentrations were 8-30 times as high as the respective serum concentrations. No significant differences between the meperidine concentration in the gastric fluid were found among the three treatments, MA, C, or ASP, versus control. Meperidine serum concentrations differed significantly versus control on two times for MA and on one time for ASP. The elimination half-life [h] and the mean residence time [h] differed significantly in the MA treatment (1.15 +/- 0.02 h and 1.23 +/- 0.24 h) versus control (1.90 +/- 0.19 h and 2.00 +/- 0.15 h).(ABSTRACT TRUNCATED AT 250 WORDS)

To study the disinfecting effect of the lubricant Instillagel for the urethra in children, bacteriologic tests of the urethra were performed immediately before and after cystoscopy in 100 children. In addition urine cultures were taken during and after cystoscopy. The culture of the urethra before cystoscopy was positive in 75 children. For 46 of these children the second culture after cystoscopy plus Instillagel appeared to be sterile while in 20 children a considerable reduction in the number of bacteria was found. Out of 92 children with an initially sterile urine culture, the culture of the urine voided immediately after the cystoscopy appeared to be contaminated in 2 while several weeks later an infected urine was found in 5 children. From this study we may conclude that during cystoscopy Instillagel has a good disinfecting effect for the urethra in children.