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Aminoquinolines (AQs) with diaminoalkane side chains (-HNRNEt2) shorter or longer than the isopentyl side chain [-HNCHMe(CH2)3NEt2] of chloroquine are active against both chloroquine-susceptible and -resistant Plasmodium falciparum. (De, D.; et al. Am. J. Trop. Med. Hyg. 1996, 55, 579-583). In the studies reported here, we examined structure-activity relationships (SARs) among AQs with different N, N-diethyldiaminoalkane side chains and different substituents at the 7-position occupied by Cl in chloroquine. 7-Iodo- and 7-bromo-AQs with diaminoalkane side chains [-HN(CH2)2NEt2, -HN(CH2)3NEt2, or -HNCHMeCH2NEt2] were as active as the corresponding 7-chloro-AQs against both chloroquine-susceptible and -resistant P. falciparum (IC50s of 3-12 nM). In contrast, with one exception, 7-fluoro-AQs and 7-trifluoromethyl-AQs were less active against chloroquine-susceptible P. falciparum (IC50s of 15-50 nM) and substantially less active against chloroquine-resistant P. falciparum (IC50s of 18-500 nM). Furthermore, most 7-OMe-AQs were inactive against both chloroquine-susceptible (IC50s of 17-150 nM) and -resistant P. falciparum (IC50s of 90-3000 nM).

Background: Measurements of cardiac output with the thermodilution technique add to data for clinical decision making and therefore must be valid and reliable. However, the results of studies on the accuracy of values obtained with room-temperature and iced injectates, especially in patients with high or low cardiac output, have been conflicting., Objective: To determine the effect of the temperature of the injectate (iced or room temperature) on cardiac output values obtained with the thermodilution technique in critically ill adults with known low cardiac output., Methods: A convenience sample of 50 subjects (41 men and 9 women) who had a cardiac index of less than 2.5 (calculated as cardiac output in liters per minute divided by body surface area in square meters) before the study had cardiac output measured by using a closed system and manual injections of room-temperature and iced injectates., Results: A paired t test indicated no significant difference between iced and room-temperature injectates for cardiac output (iced, 3.62 L/min; room temperature, 3.71 L/min; t = 0.99; P = .327) and cardiac index (iced, 1.95; room temperature, 1.99; t = 0.71; P = .482)., Conclusion: The findings support the practice of using room-temperature injectate to measure cardiac output in patients with low cardiac output.

Jack bean urease catalyzes the hydrolysis of thiourea with a second-order rate constant (kcat/Km) of 1.6 (+/- 0.2) x 10(-3) M-1 S-1 at pH7, 25 degrees C. This value is lower than that for urea by a factor of 3 x 10(8). The corresponding substitution of S for O in acetamide reduces the kcat/Km value by only a factor of 33. This greater reactivity of the oxo compounds than of the corresponding thiono compounds, and the tighter binding of urea (Ks = 2.9 mM) than of either the guanidinium ion (Ki = 30 mM) or thiourea (Ki = 70 mM), suggests that the substrate chalcogen (S or O) is more likely to be stabilized in the transition state by coordination to the enzyme via a neutral hydrogen-bond donor (i.e., Brønsted acid catalysis) than by coordination via one of the active-site nickel ions (i.e., Lewis acid catalysis).

A polysaccharide (MAR-10) was isolated from the aqueous extract of the plant Hyssop officinalis and examined for its activity against HIV-1 (SF strain) in HUT78 T cell line and primary cultures of peripheral blood mononuclear cells. MAR-10, in a concentration-dependent manner, inhibited HIV-1 replication as demonstrated by the inhibition of HIV-1 p24 antigen and syncytia formation. Furthermore, MAR-10 had no significant direct toxicity or effect on lymphocyte functions or CD4+ and CD8+ T cell counts. In addition, MAR-10 has broad spectrum anti-glycosidase activity. Our study demonstrates that MAR-10 contains strong anti-HIV-1 activity that may be useful in the treatment of patients with HIV-1 infection.

Medullipin I (Med I) is a vasodepressor prohormone which is continuously elaborated into the renal venous effluent (RVE) of isolated rat kidneys perfused under high pressure. We have improved the yield of Med I by substituting saline for the albumin perfusate previously reported; and considerably improved refinement by directly fractionating the crude lipid extract of the RVE with high pressure liquid chromatography. The results show that Med I, as defined by previous physiologic and pharmacologic criteria, is not a single molecule. The 3 Class I medullipins described here are distinguished by subtle or overt differences in polarity and biologic activity.

Objective: To determine whether secretion of medullipin I by the kidney is dependent on the cytochrome P-450 enzyme system in Sprague-Dawley and spontaneously hypertensive rats (SHR)., Methods: Isolated kidneys from Sprague-Dawley rats were perfused with 5% human albumin gassed with 95% O2 and 5% CO2 at 185 mmHg. The resultant renal venous effluent was tested in the SHR for medullipin I-type vasodepressor activity. The kidneys were then treated with ketoconazole, and inhibitor of the cytochrome P-450 enzyme system. After rinsing with 50 ml saline, the last 10 ml of which was saved for a control test (see below), the kidneys were reperfused with 50 ml human albumin and the resultant renal venous effluent was tested for vasodepressor activity. One milliliter of the saline rinse was administered to the SHR and the preketoconazole renal venous effluent was administered after 15 min. The medullipin I-type vasodepression occurred. Thus, inhibition of vasodepression after ketoconazole treatment was not due to residual ketoconazole in the post-treatment renal venous effluent., Results: Treatment of isolated kidneys with ketoconazole prevented secretion of medullipin I which had been induced by 5% human albumin., Conclusion: The cytochrome P-450 enzyme system is involved in two major metabolic steps of the medullipin system: synthesis of medullipin I by the kidney and conversion of medullipin I to medullipin II by the liver as shown previously.

Previously we reported a case of persistent hypotension associated with hypermedullipinemia (Blood Pressure 1992; 1:138-148). The hypermedullipinemia appeared to result from the autonomous secretion of medullipin I (Med I) by renomedullary interstitial cells (RIC's) in the patient's remaining endstage kidney. The patient subsequently died. At autopsy, the remaining kidney contained a yellow mass (1 x 1 x 0.5 cm) consisting of adipocytes and RIC's, termed a lipomedullipinoma. This mass was extracted and chromatographed by procedures known to yield Med I. Med I was identified following these procedures. Renal tissue outside the yellow mass failed to yield Med I. It appears that the hypermedullipinemia of this case resulted from autonomous, hypersecretion of Med I by the lipomedullipinoma.

Several bisphosphonates were examined as inhibitors of yeast GPD (glyceraldehyde-3-phosphate dehydrogenase, EC 1.2.1.12) and PGK (phosphoglycerate kinase, EC 2.7.2.3). The phosphonomethyl analog of 2-deoxy-1,3-bisphosphoglycerate (i.e., 2-oxo-1,5-bisphosphonopentane, 2-oxo-PC5P) is a good inhibitor of PGK (Ki = 0.2 +/- 0.08 mM at pH 8.5, 27 degrees C) and a poor inhibitor of GPD (Ki = 20 +/- 1 mM, pH 8.5). The shorter, butane, analog (2-oxo-PC4P) binds more tightly to PGK (Ki = 84 +/- 6 microM), and about equally well to GPD, as does 2-oxo-PC5P. The 2-oxo-bisphosphonates bind to PGK more tightly (by approx. 4 kJ/mol) than do the corresponding non-carbonyl analogues (1,4-bisphosphonobutane and 1,5-bisphosphonopentane).

A new syndrome is described in a patient with advanced renal insufficiency. This consists of severe and persistent hypotension causing weakness but associated with a clear mental status. Also present is evidence for decreased vascular reactivity. The hypotension was not orthostatic. The hypotension was associated with a circulating vasodepressor substance having the characteristics of medullipin 1. The medullipin appears to have been derived from the remaining right kidney. Hypotension existed despite the presence of major prohypertensive mechanisms, including an endstage kidney, hyperreninemia and hyperaldosteronemia. It is likely that hypotension due to hypermedullipinemia is an entity occurring in the human being.

Objective: To test the hypothesis that the secretion of medullipin I by the kidney involves an oxidative step., Design: Medullipin I is secreted by kidney renomedullary interstitial cells and is converted to medullipin II by the liver. Medullipin I can be derived from the kidney in the renal venous effluent by perfusing normal rat kidneys with 95% O2- 5% CO2 at an elevated pressure (180 mmHg). To evaluate whether the secretion of medullipin I involves an oxidative step normal rat kidneys were perfused at an elevated pressure in the presence of O2, in the absence of O2 and after treatment of the kidneys with a powerful antioxidant., Methods: Normal rat kidneys were perfused with 5% albumin bubbled with O2-CO2 at 180 mmHg. This was the control procedure for each of the three approaches. In approach (1), the kidneys were perfused with 5% albumin bubbled with N2. In approach (2), the kidneys were perfused with 'blood' treated with carbon monoxide. In approach (3), the kidneys were treated with the antioxidant butylated hydroxytoluene then perfused with 5% albumin bubbled with O2-CO2. Each perfusate was tested for medullipin I activity by rapid intravenous injection into the SHR., Results: All three approaches, which exclude the action of molecular O2, prevented the secretion of medullipin I by the kidneys., Conclusion: The secretion of medullipin I by the kidneys involves an oxidative step.

Vasodepressor prostaglandins (PGs), PGE2, PGI2, and medullipin I (Med I) are synthesized in the kidney. These vasodilator substances are thought to be involved in the antihypertensive function of the kidney. At issue is whether there are biologic differences between the vasodilator PGs and Med I. Two separate studies have shown that Med I's vasodepressor action is inhibited by four procedures: mixing with Tween 20; treatment with n-butyl boronic acid; treatment of the assay animal with SKF 525A, an inhibitor of cytochrome P-450; and removing the liver from the circulation. These same procedures were applied to the vasodilator PGs. All four failed to inhibit the vasodepressor action of the PG's. It is concluded that Med I and vasodilator PGs of the kidney are separate and distinct biologic entities.

Pseudomyxoma peritonei is a clinical diagnosis of massive abdominal swelling by a gelatinous material, produced usually from an ovarian or appendiceal primary. It is a rare entity that is usually histologically benign but behaves clinically in a malignant fashion with recurrent growth, although not demonstrating histological stromal invasion. The disease remains localized to the peritoneal cavity and the clinical course is one of repeated episodes of intestinal obstruction caused by extrinsic compression that seem only to be relieved by surgical debulking. Variable responses have been obtained with adjuvant chemo-, radio- and immunotherapy, but these isolated responses are unable to be reproduced and so there is no accepted adjuvant treatment for this disease.

A recently developed theory of the light scattering by random coils undergoing random scission is applied to the digestion of hyaluronate by hyaluronidase. The time dependence of the scattered light from solutions undergoing digestion was monitored. Working at a high angle with high molecular weight hyaluronate allowed the use of a powerful approximation for determining initial velocities and the Henri-Michaelis-menten coefficients, without explicit knowledge of the hyaluronate molecular weight, radius of gyration, second virial coefficient, or polydispersity. Effects due to a molecular weight dependent second virial coefficient and to non-Gaussian behavior are briefly considered. Assays were performed over nearly two orders of magnitude in substrate concentration. The initial velocities are compared with those obtained by a standard reducing sugar assay, which was performed on identical samples. The main advantages of the light scattering assay procedure over the more traditional assays are that many relatively high-precision data points can be quickly and automatically collected with simple apparatus, and that the technique is most sensitive for the initial period of digestion, where the other assays are least sensitive. The shapes of the scattering curves also provide evidence that hyaluronate in these solutions is not a stable double strand and that the hyaluronidase cleaves bond randomly. The curves also indicate that enzyme deactivation occurs, which accounts for the lower velocities yielded by the slower reductimetric assay, which is measured over longer initial periods.

The experimental evidence reviewed in this article suggests that the kidneys may have an additional function in regulating blood pressure besides their role in controlling both blood volume by urine formation and the relative state of vasoconstriction by the renin-angiotensin system. That is, the kidneys may have an additional influence upon the vasculature of a hormonal vasodilating system. The interstitial cells of the renal medulla appear to be mediating this activity and lipid compounds have been extracted from the renal medulla which display depressor activity. One such compound, the antihypertensive polar renomedullary lipid (APRL), has been demonstrated to consist of specific alkyl ether analogues of phosphatidylcholine. The vascular responses to these compounds include vasodilation of both arterioles and venules, rapid lowering of arterial blood pressure with little or no tachycardia, increased depressor activity in hypertensive animals, and blockade of vascular smooth muscle alpha 1-adrenergic receptors. Most recently, APRL and a synthetic analogue, 1-0-octadecyl-2-acetyl-sn-glycero-3-phosphorylcholine, have been used to demonstrate alpha-adrenergic receptor blockade on a smooth muscle cell line (DDT1) by radioligand assays. This action may be due to the insertion of these compounds into cell membranes causing subsequent steric interactions and blockade of the alpha-adrenergic receptor.

The second-order rate constants (kcat/Km) for the beta-glucosidase-catalyzed hydrolysis of aryl beta-D-glucopyranosides show a bell-shaped dependence of pH. The pKas that characterize this dependence are 4.4 (delta Hion approximately equal to 0) and 6.7 (delta Hion approximately equal to 0). In D2O these pKas are increased by 0.5 (+/- 0.1) unit, but there is no solvent isotope effect on the pH-independent second-order rate constant. Nath and Rydon [Nath, R. L., & Rydon, H. N. (1954) Biochem. J. 57, 1-10] examined the kinetics of the beta-glucosidase-catalyzed hydrolysis of a series of substituted phenyl glucosides. We have extended this study to include glucosides with phenol leaving groups of pKa less than 7. Brønsted plots for this extended series were nonlinear for both kcat/Km and kcat. Brønsted coefficients for those compounds with leaving groups of pKa greater than 7 (for kcat/Km) or pKa greater than 8.5 (for kcat) were nearly equal to -1.0, indicating substantial negative charge buildup on the leaving group in the transition state. The nonlinearity indicates an intermediate in the reaction. This was confirmed by partitioning experiments in the presence of methanol as a competing glucose acceptor. A constant product ratio, [methyl glucoside]/[glucose], was found with aryl glucoside substrates varying over 16,000-fold in reactivity (V/K), indicative of a common intermediate. Viscosity variation (in sucrose-containing buffers) was used to probe the extent to which the beta-glucosidase reactions are diffusion-controlled.(ABSTRACT TRUNCATED AT 250 WORDS)

The ANRL was derived from the renal venous effluent as the kidney exerted its nonexcretory antihypertensive function. This was made possible by three developments: (1) improvement in the extraction of ANRL from fresh renal medulla; (2) the fact that purified ANRL caused an acute vasodepressor effect (acted as a vasodilator); and (3) experience with unclipping the one-kidney, one-clip hypertensive rat. Unclipping after an anastomosis between the ureter and the vena cava caused the MAP to return to normal levels in an average of 20 hr. At an average of 5 hr, when the MAP had dropped an average of 34 mm Hg (from approximately 190), an exchange infusion was started and blood was collected from the renal vein. The plasma was separated, lyophilized, and extracted for total lipids. The lipids were subjected to two TLC procedures and tested for vasodepressor activity. Renal venous effluent, under those conditions, yielded a considerable amount of vasodepressor lipid that was similar to that derived from fresh renal medulla. Controls (normal, nephrectomized, and hypertensive animals) yielded little or no such lipid. Indomethacin did not interfere with the derivation of the vasodepressor lipid. As the MAP was lowered and the ANRL-like lipid appeared in the renal venous blood, the RICs degranulated. The RICs appear to be the source of the antihypertensive lipid.

The growth inhibitory effect of 6-hydroxydopa, a cytotoxic analog of L-dopa, was studied in melanotic and amelanotic Cloudman melanoma, mouse fibroblast L929 and Chinese hamster ovary cells. A marked sensitivity of pigmented cells to 6-hydroxydopa was observed with a 10-fold increase in sensitivity of pigmented cells. Sensitivity correlated with the capacity of cells to incorporate radiolabeled exogenous L-dopa. The drug affected primarily DNA and RNA synthesis with greater inhibition observed in pigmented cells than the nonpigmented control cells. The mechanism of action may involve interaction with the melanocyte specific enzyme, tyrosinase, as a false substrate.

The specificity of induced conformational changes and of the probes used to detect them has been investigated in yeast glyceraldehyde-3-phosphate dehydrogenase. Cyanylation of the active-site SH groups in two of the four identical subunits of glyceraldehyde-3-phosphate dehydrogenase has no effect on reactivity of the unmodified SH groups toward the cyanylating reagent (2-nitro-5-thiocyanogenzoic acid, NTCB) but results in total loss of catalytic activity. Cyanylation of the dicarboxamidomethylated enzyme was four orders of magnitude slower than with the unmodified enzyme in contrast to cyanylation of the dicyanylated enzyme. Cyanylation by NTCB as well as alkylation by iodoacetate and acylation with beta-(2-furyl)acryloyl phosphate are enhanced in the presence of NAD+ while alkylation by iodoacetamide is inhibited by NAD+. In the absence of NAD+, hydrolysis of the acylated enzyme is faster than phosphorolysis while the reverse is true in the presence of NAD+. NAD+ accelerates hydrolysis of the 3-phosphoglyceroylated enzyme about 60-fold but decreases the rate of hydrolysis of the furylacryloylated enzyme by a factor of 17. Other examples of the specificity of the induced conformational changes and the probes are described. The conformational changes induced by NAD+ make the protein specifically reactive toward its physiological substrates and less reactive toward extraneous competing compounds.

Chorioangiomas are usually small and found by careful sectioning of the placenta. In recent years ultrasonography has been used in antenatal diagnosis. Multiple and large tumors (greater than 5 cm) are associated with increases in fetal morbidity and mortality. We treated a woman who had severe polyhydramnios and a large chorioangioma with vascular channels in the tumor confirmed by Doppler studies. Daily fetal breathing was used to monitor fetal well-being.

A variety of reversible inhibitors of sweet almond beta-glucosidase were examined. These included simple sugars and sugar derivatives, amines and phenols. With respect to the sugar inhibitors and, indeed, the various glycoside substrates, the enzyme has what can be considered a "relaxed specificity". No single substituent on glucose, for example, is essential for binding. Replacement of a hydroxyl group with an anionic substituent reduces the affinity while substitution with a cationic (amine) substituent enhances the affinity. Amines, in general, are good inhibitors, binding more tightly than the corresponding alcohols: pKiRNH3+ = 0.645pKiROH + 1.77 (n = 9, r = 0.97). The affinity of a series of 10 primary amines was found to be strongly influenced by substituent hydrophobicity: pKi = 0.52 pi + 1.32 (r = 0.95). The major binding determinant of the glycoside substrates is the aglycon moiety. Thus, the Ki values of phenols are similar in magnitude to the Ks values of the corresponding aryl beta-glucoside. The pH dependence for the inhibition by various phenols indicates that it is the un-ionized phenol which binds to the enzyme when an enzymic group of pKa = 6.8 (+/- 0.1) is protonated. The affinity of the phenol inhibitor is dependent on its basicity with a Brønsted coefficient for binding of beta = -0.26 (n = 14, r = 0.98). The pH dependence of the binding of two particularly potent beta-glucosidase inhibitors was also examined. 1-Deoxynojirimycin (1,5-dideoxy-1,5-imino-D-glucitol) has a pH-corrected Ki = 6.5 microM, and D-glucono-1,5-lactam has a pH-corrected Ki = 29 microM.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of pure synthetic PAF (1-0-alkyl-2-acetyl-sn-glycero-3-phosphorylcholine) was studied in human platelets. PAF (0.2--2.0 micrograms/ml) produced a dose-dependent aggregation in human platelet-rich plasma (PRP) or platelet suspension obtained by gel-filtration (GFP). In addition, PAF (0.8 microgram/ml) induced secretion of 14C-serotonin (45% +/- 10%; mean +/- SD, n = 9) and platelet factor 4 (PF4) (12.89 +/- 3.81 micrograms/10(9) platelets; n = 9) in PRP. Similar results were obtained in GFP. Aggregation and release of 14C-serotonin and PF4 were inhibited by the metabolic inhibitors 2-deoxyglucose (16.7 mM) and antimycin-A (8.3 micrograms/ml), by the membrane-active drugs mepacrine (10 microM) and chlorpromazine (0.025 mM), by PGI2 (5.34 nM), which elevates intracellular c-AMP, by indomethacin (10 microM) or aspirin (100 microM). The ADP scavengers, creatine phosphate and creatine phosphokinase (CP/CPK), inhibited the second wave of aggregation but not secretion. These data suggest that the major effect of PAF on human platelets is mediated through the cyclo-oxygenase pathway and not through a third pathway.

A reno-hepatic axis of blood pressure (BP) control has been proposed primarily by two developments: the lag period between the injection of the antihypertensive neutral renomedullary lipid (ANRL) into the vena cava and the beginning of the drop of the BP is significantly reduced by injection of this lipid into the portal vein; and a reno-portal shunt (RPS) accelerates markedly the drop of the BP following unclipping the Goldblatt hypertensive rat. In the present studies, three maneuvers were performed: perfusion of the isolated, blood-drained rat liver with plasma containing ANRL and delivery of the plasma into the jugular vein of a hypertensive rat; injection of captopril, establishment of a RPS of a hypertensive rat (one-kidney, one-clip) followed by unclipping; and establishment of a RPS in a normal rat. The blood-drained liver was capable of decreasing significantly the lag period of ANRL. Captopril did not potentiate the antihypertensive action of the kidney when RPS was coupled with unclipping. Following the RPS, the BP of the normal rat dropped modestly over a 3-hour period. These results further support the concept of reno-hepatic axis of BP control.

The glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase catalyzes the oxidative phosphorylation of D-glyceraldehyde 3-phosphate. A variety of phosphonates have been shown to substitute for phosphate in this reaction [Gardner, J. H., & Byers, L. D., (1977) J. Biol. Chem. 252, 5925--5927]. The dependence of the logarithm of the equilibrium constant for the reaction on the pKa2 value of the phosphonate is characterized by a Brłnsted coefficient, betaeq, of approximately 1. This represents the sensitivity of the transfer of the phosphoglyceroyl group between the active-site sulfhydryl residue (in the acyl-enzyme intermediate) and the acyl acceptor on the basicity of the acyl acceptor. Molybdate (MoO42-) can also serve as an acyl acceptor in the glyceraldehyde-3-phosphate dehydrogenase catalyzed reaction. The second-order rate constant for the reaction with molybdate is only approximately 12 times lower than the reaction with phosphate even though the pKa2 of molybdate is 3.1 units lower than the pKa2 of phosphate. The immediate product of the molybdate reaction is the acyl molybdate, 1-molybdo-3-phosphoglycerate. The acyl molybdate, like the acyl arsenate (the immediate product of the reaction when arsenate is the acyl acceptor), is kinetically unstable. At pH 7.3 (25 degrees C), the half-life for hydrolysis of the acyl molybdate, or the acyl arsenate, is less than 2.5 s. Thus, hydrolysis of 1-molybdo- and 1-arseno-3-phosphoglycerate is at least 2000 times faster than hydrolysis of 1,3-diphosphoglycerate under the same conditions. Glyceraldehyde-3-phosphate dehydrogenase has a fairly broad specificity for acyl acceptors. Most tetrahedral oxy anions tested are substrates for the enzyme (except SO4(2-) and SeO4(2-)). Tetrahedral monoanions such as ReO4- and GeO(OH)3- are not substrates but do bind to the enzyme. These results suggest the requirement of at least one anionic site on the acyl acceptor required for binding and another anionic group on the acyl receptor required for nucleophilic attack on the acyl enzyme.

Bone marrow from patients with acute myelogenous leukemia (AML), acute myelomonocytic leukemia (AMML), chronic myelogenous leukemia (CML), preleukemia, and from healthy volunteers was cultured using a recently developed liquid diffusion technique. Differential and viable cell counts and 3H-thymidine labeling indices were performed at intervals up to 30 days. Differentiation was assessed morphologically by light and electron microscopy, histochemically, and by functional tests for phagocytosis and the presence of surface receptors for IgG. Colony-stimulating activity (CSA) was assayed against normal human bone marrow by the agar colony technique. In acute leukemia cultures, viable cell counts usually fell within the normal range. However, most AML cells failed to demonstrate significant maturation in vitro, and did not produce detectable CSA. In AMML cultures, maturation was defective but some differentiated macrophages were observed and the cells produced high concentrations of CSA. Preleukemic cultures demonstrated normal growth but maturation was impaired as evidenced by a high percentage of immature cells during the first 7 days. CML cultures showed abnormally high growth capacity resulting in viable cell counts 2-3 times normal. In the chronic phase of CML, maturation was qualitatively normal and the cells produced CSA. With the onset of blast transformation, maturation became abnormal but growth remained high. These studies support a concept of AML as a primary defect in cellular maturation and of CML as a primary abnormality of proliferation. The production of CSA by neoplastic cells relates to the degree of monocyte-macrophage differentiation within the leukemic population. Human preleukemia is characterized by a failure of normal maturation in vitro.

Medullipin I causes a delayed onset depressor response when injected intravenously into rats. The glyceryl compounds selachyl alcohol (SA) and monoolein (MO) cause similar vasodepression. The neutral lipid 1-O-hexadecyl-2-acetyl-sn-glycerol (HAG) was suggested by Blank et al to be medullipin I (Med I, formerly ANRL). Biologic comparisons were made between Med I and various glyceryl compounds, including SA, MO, HAG, alkyl glyceryl ethers of phosphatidyl choline (termed APRL by us), diacylated SA, and the n-butyl boronic acid derivative of SA and MO. The n-butyl boronic acid derivative of Med I also was evaluated. The delay in onset of the depressor response to Med I was reduced by the injection of Med I into the portal vein; that of SA and MO was not. Med I, SA, and MO were activated by the liver, while APRL and HAG were not. Tween 20 inhibited Med I, SA, and MO, but not APRL and HAG. Proadifen (SKF 525A) inhibited Med I, but not SA and MO. The n-butyl boronic acid derivatives of SA, MO, and Med I were inactive. Med I, like SA and MO, appeared to have two hydroxyl groups in close proximity. It was concluded that Med I is neither HAG, APRL, SA, nor MO.

In a study of the effect of L-dopa, an intermediate in the biosynthesis of the pigment melanin, on the growth of human and murine melanoma cells a highly selective inhibition of growth was observed for pigmented cell lines (S91A and human melanoma) as compared to the nonpigmented control cells (amelanotic melanoma S91B, mouse fibroblast L929, and Chinese hamster ovary). There was a correlation between toxicity and the extent of incorporation of radioactively labeled L-dopa by each line.

Selachyl alcohol (SA) is a mono-oleyl glyceryl ether. It has certain biologic activities similar to those of the antihypertensive neutral renomedullary lipid (ANRL) derived from the renal papilla and its renomedullary interstitial cells (RIC). These include a vaso-depressor effect following bolus injection and a requirement for hepatic activation for the development of biological activity. In view of this similarity to ANRL, it appeared worthwhile to test the antihypertensive action of SA when given via the GI tract. Accordingly, pure SA was given either by gavage or by tube into the stomach or duodenum of one-kidney, one-clip hypertensive rats (5-10 mg per dose). The role of hepatic activation was demonstrated by comparing the BP response to bolus injection of SA and ANRL with and without the presence of an intact circulation to the liver. Administration of SA via the GI tract resulted in a significant decline in BP without tachycardia or weight loss. In the absence of a circulation to the liver, neither SA nor ANRL was active. SA appears to be an effective antihypertensive agent when given via the GI tract.

Transplants and lipid extracts of the same monolayer tissue culture of renomedullary interstitial cells from murine renal medulla exerted a similar antihypertensive action in rats having hypertension of the sodium-volume-dependent-type. The antihypertensive action resembled that caused by lipid extracts of rabbit renal medulla and extracts of lapine renomedullary interstitial cells grown in tissue culture. The recession of the arterial pressure of the hypertensive animals usually occurred slowly and steadily to a maximum within 6 to 12 hours. On occasions, a substantial acute depressor effect preceded the slow and steady decline of the pressure. As the pressure was lowered, there was either minimal or no change in the pulse rate. The lowering of the hypertensive pressure before there was vascularization of the transplant appears to support the view that the transplanted cells secreted and/or liberated an antihypertensive substance(s) that seeped out and was absorbed by nearby capillaries and/or lymphatics and circulated and acted in the manner of a hormone. The extracted and purified lipid from the same cells as used for transplantation is proposed as a candidate for such hormonal action. Evidence is presented that minimizes the possibility of the classic renomedullary prostaglandins as this antihypertensive lipid. The findings add support to the concept that the kidney exerts a hormonal antihypertensive action that opposes the well known hormonal prohypertensive renal actions.

Over 25 nitrogen-containing heterocycles were tested as inhibitors of sweet almond beta-glucosidase (EC 3.2.1.21). Among the most potent of these are some imidazole derivatives. The pH dependence indicates that the unprotonated inhibitor binds most tightly to the catalytically active species of the enzyme. This is analogous to the situation with 1-deoxynojirimycin where the permanently cationic species, N,N-dimethyl-1-deoxynojirimycin, binds at least two orders of magnitude less tightly to the enzyme than does the unprotonated 1-deoxynojirimycin. The binding of imidazole derivatives show a general tendency of increasing affinity with increasing basicity (beta approximately 0.4). One derivative which shows a significant positive deviation from this correlation (- log Ki vs. pKa) is 4-phenylimidazole. 4-Phenylimidazole is one of the most potent reversible inhibitors of beta-glucosidase with a pH-independent Ki = 0.8 microM. It is also fairly specific for beta-glucosidase, binding at least three orders of magnitude less tightly to any of the other exoglycosidases tested. This inhibitor combines, in a mono-molecular species, the binding affinities of benzene, which binds at the hydrophobic aglycone binding site, and imidazole, which binds at the sugar binding site of beta-glucosidase. The binding energy of 4-phenylimidazole can be attributed to the sum of the intrinsic binding energies of the phenyl and imidazole moieties. Thus, there is no significant entropic advantage of combining the component parts of phenylimidazole in a single species. This indicates that there is no significant uncompensated entropy loss upon binding of either benzene or imidazole to the enzyme. Nevertheless, the additivity of binding energy, even in the absence of an entropic advantage, results in the most powerful known inhibitor of the enzyme.

The spontaneous inactivation of yeast glyceraldehyde-3-phosphate dehydrogenase was found to fit a simple two-state model at pH 8.5 and 25 degrees. The first step is a relatively rapid dissociation of the tetramer to dimers with the equilibrium largely in favor of the tetramer. In the absence of NAD+ the dimer inactivates irreversibly. The apoenzyme is quite stable with a half-life for complete activity loss proportional to the square root of the enzyme concentration. Perturbances of the protein structure (by pH, ionic strength, and specific salts), which have no effect on the tetrameric state of the molecule, result in an alteration of the cooperativity of NAD+ binding, the reactivity of the active-site sulfhydryl group, and the catalytic activity of the enzyme. Covalent modification of two of the four active-site sulfhydryl groups has profound effects on the enzymic activity which are mediated by changes in the subunit interactions. Sedimentation analysis and hybridization studies indicate that the interaction between subunits remains strong after covalent modification. Under normal physiological and equilibrium dialysis conditions the protein is a tetramer. Equilibrium dialysis studies of NAD+ binding to the enzyme at pH 8.5 and 25 degrees reveal a mixed cooperativity pattern. A model consistent with these observations and the observed half-of-the-sites reactivity is that of ligand induced sequential conformational changes which are transferred across strongly interacting subunit domains. Methods for distinguishing negatively cooperative binding patterns from mixtures of denatured enzyme and multiple species are discussed.

1. Allogenic transplants of cultured renomedullary interstitial cells exert a powerful anti-hypertensive action. The blood pressure of hypertensive animals usually drops slowly over 8-12 h whereas the pulse is unchanged or reduced. 2. Lipids derived from the cultured cells exert a similar anti-hypertensive action. 3. The anti-hypertensive action of transplanted cultured cells almost certainly results from the secretion of a substance(s) that acts in the manner of a hormone. The tissue culture lipid is a prime candidate hormone. 4. The relationship of the kidney to the hypertensive state is considered to entail pro- and anti-hypertensive actions. The pro-hypertensive actions include (a) activation of the renal pressor system (mainly renin-angiotensin), (b) failure to prevent sodium and fluid overloading because of either an injured or absent kidney or the excessive action of mineralocorticoids (mainly aldosterone). The antihypertensive actions of the kidney include (a) the relief of sodium and fluid overloading through diuresis-natriuresis and (b) the action of the reno-medullary interstitial cell hormone (the antihypertensive renomedullary hormone).

Four types of lipids with potential antihypertensive properties have recently been derived from the kidney. These consist of prostaglandins (PG), a renin inhibitor, a neutral lipid, and alkyl ether analogs of phosphatidylcholine. PGI2, mostly renocortical, and PGE2, mostly renomedullary, may aid the antihypertensive function of the kidney by decreasing renal vascular resistance and shunting blood toward the juxtamedullary zone and the renal papilla, where the renomedullary interstitial cells (RIC) are located. Several analogs of the renin inhibitor are available. The neutral lipid is a natural product derived from fresh renal medulla and from RIC grown as monolayer tissue culture. The alkyl ether analogs of phosphatidylcholine (formerly designated as the antihypertensive polar renomedullary lipid or APRL) are orally active vasodilators. They cause a prolonged depressor effect due to a decrease in peripheral vascular resistance. The latter may be partly due to alpha-adrenergic antagonism.