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Summary A model whereby a known number of coagulase-negative staphylococci were packed into capillary tubes and implanted into the peritoneal cavity of mice proved to be a satisfactory method for generating abscesses that could be easily removed free of extraneous host tissue, and that permitted measurement of the survival of the organisms and accumulation of lipid in the lesion. Strains of S. epidermidis, S. schleiferi and S. lugdunensis, differing in their ability to produce fatty acid modifying enzyme (FAME) and lipase, were packed into either glass or plastic capillary tubes and used to generate abscesses. Abscesses produced by S. aureus served as comparators. Lipids accumulated within the abscesses caused by S. aureus in the same manner as previously described for the organism inoculated without tubes.1 Lipids also accumulated within abscesses produced by all the coagulase-negative staphylococci, but the rate of accumulation was slower and the lipid droplets were smaller than seen with S. aureus. The mobilisation of lipid did not differ in response to cocci in plastic or glass tubes. Strains of S. epidermidis and S. schleiferi producing FAME and lipase were better able to survive within abscesses than strains unable to produce these enzymes. However, FAME and lipase production did not appear to be the sole determinants of survival within abscesses. Regardless of whether they produced FAME and lipase, the two S. epidermidis strains were significantly better able to survive within plastic tubes than in glass tubes. No such difference was seen with S. aureus between plastic and glass tubes.

Serotonin1A receptor agonists, 8-hydroxy-2-(di-n-propylamino)tetralin and 10-methyl-11-hydroxyaporphine, inhibited electrical stimulation-induced contraction of the guinea-pig ileum. These agonists also inhibited the pressor and tachycardiac responses to low frequency (0.25 Hz) but not to high frequency (2.0 Hz) electrical stimulation of the sympathetic nervous system in pithed rats. Serotonin1B receptor agonist RU 24969 inhibited pressor and tachycardiac responses to both low and high frequencies of stimulation in pithed rats. In the cat nictitating membrane, serotonin1A receptor agonists did not alter contractions elicited by electrical stimulation (0.1–3.0 Hz). Serotonin not only contracted the cat nicitating membrane but also facilitated contractile responses to low frequency (0.1 – 1.0 Hz) stimulation. The contractile effect of serotonin in the cat nictitating membrane was blunted by bretylium, methysergide, and ketanserin, but not by metoclopramide. The facilitatory effect of serotonin was antagonized by methysergide, but not by ketanserin, pindolol, propranolol, or metoclopramide. These results suggest that serotonin1A receptors modulate autonomic neurotransmission in the guinea-pig ileum and pithed rats, but not in the cat nictitating membrane. Serotonin contracts the cat nictitating membrane via serotonin2 subtypes, while facilitating stimulated contractile responses through the serotonin1-like receptors.Key words: guinea-pig ileum, pithed rats, nictitating membrane, serotonin receptors.

Immunization with recombinant S. pneumoniae neuraminidase NanA (rNanA) resulted in a significant reduction in pneumococcal colonization in the chinchilla model. The bacteria were eliminated from the nasopharynx 1 week earlier than that from the control cohort. Our data suggest that rNanA affords protection against pneumococcal nasopharyngeal colonization.

Streptococcus pneumoniae neuraminidase has been implicated as a virulence factor in the pathogenesis of pneumococcal otitis media. In this study, native neuraminidase was partially purified from cultures of S. pneumoniae by serial chromatography with DEAE-Sepharose and Sephacryl S-200. Recombinant neuraminidase, a 3,038-bp fragment of the neuraminidase A ( nanA ) gene, was cloned into the pET-28b vector and then expressed at high levels in Escherichia coli . Chinchillas were immunized subcutaneously with either the gel-purified native or recombinant neuraminidase, and all responded with elevated titers of antineuraminidase antibody in serum. Immunization with neuraminidase resulted in a significant reduction in nasopharyngeal colonization as well as in the incidence of otitis media with effusion. These data demonstrate for the first time that neuraminidase affords protection against S. pneumoniae nasopharyngeal colonization and experimental otitis media.

Phase variation in the colonial opacity phenotype of Streptococcus pneumoniae has been implicated as a factor in bacterial adherence, colonization, and invasion in the pathogenesis of pneumococcal otitis media (OM). The purpose of this study was to determine whether S. pneumoniae opacity variants influence the induction of gene expression for proinflammatory mediators in vivo using the rat model of OM. Both the opaque and transparent phenotype variants induced a significant up-regulation in gene expression for interleukin-1α (IL-1α), IL-1β, IL-6, IL-10, tumor necrosis factor alpha, and inducible nitric oxide synthase (iNOS) compared to saline sham-inoculated controls at both 4 and 24 h postinoculation ( P < 0.05 in all cases). Furthermore, whereas a significant difference in gene expression was evident for only IL-6 (greater following challenge with the opaque variant) and IL-1β (greater following challenge with the transparent variant) at 4 h, by 24 h the opaque variant cohort demonstrated a significant increase in gene expression for IL-1α, IL-1β, IL-6, IL-10, and iNOS relative to animals inoculated with the transparent phenotype variant ( P < 0.05 in all cases). Enzyme-linked immunosorbent assay results confirmed the gene expression data as determined by real-time PCR. Moreover, the concentrations of the opaque variant in the middle ear lavage fluid were a full log higher than those of the transparent variant. The aforementioned results indicate that the opaque phenotype variant is more efficient at survival and multiplication within the middle ear space, resulting in the accumulation of more inflammatory cells and the enhanced expression and production of inflammatory mediators. However, when the data were normalized to account for differences in middle ear bacterial titers, it became apparent that the transparent variant of S. pneumoniae is a more potent inducer of inflammation, triggering the accumulation of more inflammatory cells and substantially greater fold increases in the expression and production of inflammatory mediators. Data from this study indicate that S. pneumoniae opacity variants influence the temporal mRNA expression of inflammatory mediators within the middle ear.

Real-time PCR and enzyme-linked immunosorbent assay were used to evaluate the ability of influenza A virus and Streptococcus pneumoniae opacity variants, either alone or in combination, to induce cytokine and chemokine genes in primary cultures of human middle ear epithelial (HMEE) cells. Following treatment with influenza A virus, the induction of gene expression, which occurred in a dose- and time-dependent manner, was strong for macrophage inflammatory protein 1α (MIP-1α) and MIP-1β; moderate for tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and IL-8; and weak for IL-1β and monocyte chemotactic peptide 1 (MCP-1). Except for TNF-α, all the gene products were detected in the cell culture supernatants. In contrast, infection of HMEE cells with S. pneumoniae alone induced low levels of mRNA expression of MIP-1α and MIP-1β and did not significantly induce the transcription of the other cytokines and chemokines examined. However, both S. pneumoniae opacity variants increased mRNA expression of MIP-1α, MIP-1β, IL-6, and MCP-1 in HMEE cells activated by a prior influenza A virus infection compared to levels in cells treated with either agent alone. Up-regulation of IL-6, IL-8, and MCP-1 mRNA expression and production by the virus in combination with opaque S. pneumoniae was two- to threefold higher than that induced by the virus combined with the transparent S. pneumoniae variant. These data indicate that the activation of HMEE cells by influenza A virus enhances the induction of cytokine and chemokine gene transcripts by S. pneumoniae and that this effect appears to be most pronounced when S. pneumoniae is in the opaque phase.

The most common genetic aberration seen in nonpapillary renal cell carcinoma is believed to be the loss or inactivation of allelic material on the short arm of chromosome 3 (3p). Two patients underwent nephrectomy at our institution, each initially receiving a histologic diagnosis of renal cell carcinoma. Molecular analysis of these tissues revealed no genetic deletion on 3p, prompting further histologic and immunohistochemical evaluation of the original specimens. These additional studies confirmed alternate histologies other than renal cell carcinoma in each case. These data suggest that the loss of genetic material on the short arm of chromosome 3 may be specific for nonpapillary renal cell carcinoma. Molecular analysis may prove to be a useful adjunct in establishing the diagnosis in renal neoplasms with equivocal histologies.

Summary Eighty-six strains encompassing 11 species of coagulase-negative staphylococci were examined for the production of fatty acid modifying enzyme (FAME) and lipase. Staphylococcus schleiferi and S. saprophyticus most closely resembled S. aureus in that 80% of the strains produced both enzymes. In contrast, no strains of S. lugdunensis and S. haemolyticus tested produced these enzymes. S. simulans was unusual in that eight of 10 strains produced FAME, but only one produced lipase. Among the other species the proportion of strains producing both enzymes ranged from 10 to 60%. Generally there was a strong correlation between FAME and lipase production.

The effect of metabolic inhibitors on norepinephrine (NE), epinephrine (E), tyramine (T), serotonin (5-HT), potassium chloride (KCl), and barium chloride (BaCl2) induced contraction was evaluated with the superfused canine dorsal metatarsal veins. ATP content was evaluated in the absence and presence of dinitrophenol, cyanide, azide, fluoroacetate, and iodoacetate. Contractile responses to vasoactive stimuli were measured before and during continuous superfusion with these inhibitors. Each of the metabolic inhibitors decreased tone and ATP. Dinitrophenol reduced the contractile responses of tibial arteries to NE, E, 5-HT, and KCl but abolished the contractile response to T and BaCl2. Cyanide and fluoroacetic acid did not significantly alter the contractile responses of tibial arteries to KCl or E but reduced the contractile responses to T, 5-HT, NE, and BaCl2. Azide reduced the responses to KCl, 5-HT, and T but not to NE, E, or BaCl2. Fluoroacetic acid reduced the responses to tyramine and serotonin but not to NE, E, KCl, or BaCl2. These findings are consistent with the conclusions that (1) vasoactive stimuli utilized different metabolic pathways to sustain their contractile response, (2) the adrenergic neuronal uptake mechanism or release process for catecholamines is more sensitive to metabolic inhibition than the contractile process, and (3) myogenic tone is more dependent on a functional oxidative metabolic pathway than the contractile responses to vasoactive stimuli.

Forty-eight patients with histologically proven hepatic cirrhosis were reviewed. Alcohol was the commonest aetiological factor. During the period of study the incidence of alcohol related cirrosis increased and the disease was seen more frequently in younger patients. This study raises important medicosocial issues.

IN a review of eleven patients with aspergilloma attending St. Vincent’s Hospital between 1973 and 1984 sarcoid osis was the commonest predisposing disease (six patients) whereas cavitary tuberculosis, previously the most frequently associated condition, was present in only two patients. All patients had a combined restrictive-obstructive pulmonary deficit with mean values for FVC, FEV, and DLCO being 49%, 35% and 46% of predicted respectively. Life threatening haemoptysis was the most significant complication, occurring in six of eleven patients. In only two of these patients was lung function sufficiently adequate to permit surgical resection and both are alive 5 and 9 years respectively following the procedure. Four patients precluded from surgery because of prohibitive lung function had emergency bronchial arteriography performed with a vitw to arterial embolisation. This was successfully achieved in three patients with prompt cessation of active major haemoptysis, but could not be performed in a fourth because of an arterial anomaly. However there were no long term survivors in this latter non-surgical group. Finally, in five untreated patients observed over a mean period of three years, no major complications developed, despite enlargement of the fungus ball in three individuals. This study demonstrates a changing pattern of underlying disease in patients with aspergilloma and illutrates the usefulness of emergency arterial embolisation in the short term management of life threatening haemoptysis.

In the anesthetized, open-chest dog, intravenous infusion 8– 13 microgram . kg-1. min-1) of a new dopamine analog 5,6-dihydroxy-2-methylaminotetralin hydrobromide (M-8) demonstrated potent coronary vasodilator properties. Blood flow to normal myocardium was increased an average of 53% above control flow values and this was associated with a decrease in coronary vascular resistance of 50%. The increase in blood flow was distributed uniformly to all portions of the myocardium (left ventricular free wall, right ventricular free wall, and septum). Subendocardial/subepicardial (endo/epi) flow ratios of the left ventricle, right ventricle, and septum were not significantly changed from control. In dog hearts subjected to acute, sudden occlusion of portions of the left anterior branch of the left coronary artery, M-8 produced a 93% increase in flow to the whole heart over postocclusion flow values. Flow to tissues of the heart made ischemic by the occlusive procedure in creased by an average of 94% during M-8 administration despite the fact that the occlusive ligatures remained in place. The vasodilation produced by M-8 was accompanied by a 22–24% increase in myocardial capillary surface area available for exchange of lipid-insoluble substances, fructose and sucrose. Hemodynamic changes associated with M-8 include a transient decrease in aortic pressure, but no change in heart rate, left ventricular (dP/dt)/P, or central venous pressure. Propranolol blocked the vasodilator activity of M-8. It was concluded that M-8 is a beta2-adrenergic receptor agonist having potent coronary vasodilator properties which also has the ability to open preexisting collateral blood flow channels and provide nutritive flow to ischemic, marginally ischemic, and normal myocardium. The cardiovascular actions of M-8 increase blood flow and oxygen delivery to the myocardium while decreasing the work and oxygen utilization by the heart, suggesting that it may have important antianginal characteristics.

The purpose of this research is the synthesis and biological evaluation of analogs of hemicholinium (HC-3). These are agents which decrease the ability of cholinergic neurones to synthesize acetylcholine. The long-range goal of this research is to develop compounds which can be used to antidote excess acetylcholine within a cholinergic synapse. Some possible approaches are 1) decrease the content of acetylcholine within the cholinergic neurone by interfering with synthesis, 2) desensitizing cholinergic receptors at post- synaptic sites, 3) decreasing the release of acetylcholine from the neurone by stabilizing the membrane or via pre-synaptic receptors which when activated will diminish the amount of acetylcholine released into the synapse. Thus far 3 agents have been prepared and evaluated for activity. Two of the agents, which are quarternary amines, approximate HC-3 in activity and the tertiary amine derivative is approximately 1/500th as active as HC-3. The latter agent is the first active non-quarternary amine to be reported. The biological assay procedures have been developed to evaluate these agents. Keywords: High performance liquid chromatography; Incubation; Caudate nucleus; Rats; Rabbits; Nerve-gastrochemisumuscle; In viro analysis; Neuro-muscular blocking; Phrenic nerve-diaphragm muscle; In vitro analysis; and Neuromuscular transmission.

Alterations in propylthiouracii (PTU) disposition after long-term use in patients treated for alcoholic liver disease (ALD) have not been reported. We gave PTU (3.3 mg / kg / day intraperitoneally) for 24 days to naive male Sprague-Dawley rats (n = 10). Clearance of PTU in these animals (1.11 ± 0.04 ml / min / kg, x ± SEM) was slightly less than in controls (1.28 ± 0.05 ml/min/kg, n = 8, P = 0.02) as a consequence of drug-induced hypothyroidism (thyroid weight in PTU-treated rats = 78.0 ± 7.2; controls = 29.6 ± 2.1 mg/kg, P < 0.01). PTU clearance in thyroidectomized rats (0.74 ± 0.04 ml/min/kg, n = 12; thyroid-stimulating hormone [TSH] = 3.3 ± 0.2; μIU/ml) was less (P < 0.01) than in sham-operated controls (1.47 ± 0.12 ml/min/kg; n = 9; TSH = 1.9 ± 0.2 μIU/ml). Neither plasma free fraction (25.0 ± 0.8%) nor apparent volume of distribution (0.72 ± 0.02 l/kg) in thyroidectomized rats differed from control values (26.4 ± 1.4%, 0.73 ± 0.02 l/kg). In ALD patients (four men; two women) tested with PTU (300 mg IV) before and after PTU therapy (150 mg b.i.d. orally) for 28 days, TSH rose from 3.6 ± 1.0 to 7.0 ± 2.6 μIU/ml (P = 0.03), but free PTU clearances rose in all patients (range 2.2% to 48.4%, P = 0.03). Free clearance before treatment was 1047 ± 131 and after treatment it was 1223 ± 139 ml/min; in 14 healthy subjects it was 1454 ± 86 ml/min. The alteration in clearance correlated positively (r = 0.91, P < 0.02) with the severity of disease as assessed by a composite clinical and laboratory index; in the more severely diseased patients there was the most alteration in both clearance and severity of ALD. There are two opposing events that determine net PTU disposition; biochemical hypothyroidism reduces PTU clearance, but improvement in hepatic function overcomes such an effect. The net increase in PTU free clearance reduces efficacy during long-term dosing. Clinical Pharmacology and Therapeutics (1982) 31, 559–563; doi:10.1038/clpt.1982.78

Antibiotic administration during surgery may be associated with hazards not normally encountered in unanesthetized subjects. We submit this contention on the bases of recent evidences in the literature and our laboratory investigations stimulated by clinical experiences. The potential dangers are not those usually associated with prolonged or repeated administration of these drugs, such as allergic responses, blood dyscrasias, nephro- and ototoxic phenomena, but rather acute states of respiratory or circulatory depression. A drug may be therapeutic under usual circumstances of administration and toxic when altered conditions prevail. The patient during surgery is indeed different from an unanesthetized subject. A number of factors could conceivably affect his response to a drug. Anesthesia poses the possibility of drug interaction with a variety of agents used for that purpose. Changes in concentrations of circulating hormones in response to surgical stress and anesthesia may modify the effect of exogenous agents. Hemodynamic changes such as

We examined the effect of ethanol on propylthiouracil (PTU) disposition in normal subjects and a rabbit. The drug is metabolized by conjugation. In six normal subjects 19.2 gm oral ethanol, taken either with or two hr after 300 mg PTU, did not change maximum concentration, time to maximum concentration, or total or free AUC of PTU. Pretreatment with ethanol and supplementation to keep blood ethanol concentration above 800 mg/l for 8 hr also did not alter PTU disposition. In the rabbit, the infusion of ethanol (8.81 mg /min) 4 hr after the beginning of an intravenous infusion of PTU 0.05 mg/min did not alter the plasma concentration profile. These results indicate that short-term ethanol dosing does not affect PTU disposition and therefore dosage adjustment is not necessary in patients who drink alcohol. Clinical Pharmacology and Therapeutics (1983) 33, 663–667; doi:10.1038/clpt.1983.90

IN spite of the simplicity of its chemical structure, details of the mode of action of acetylcholine (ACh) remain unknown. We have considered stereochemical attachment of ACh at its nicotinic and its muscarinic receptor sites, and compared these two stereochemical attachments. A logical working hypothesis, proposed by many in the past (and so far not proved), is that the flexible ACh molecule assumes different conformations and hence different molecular shapes and different interatomic distances at the nicotinic and muscarinic receptors. These conformations are imposed on the ACh molecule by the chemical and the steric nature of the receptor area.

The second volume of this overdue series continues to follow the high standards of Volume 1. The publication begins with a review of the status and recent advancements made by pharmacology in India, China, and Europe. The remainder of the volume reviews in detail major advances made in various sections of pharmacology. The 2 general areas of pharmacology, biochemical and physiological, seem to be well balanced. This balanced presentation is evidenced by the following topics: biochemical mechanism, drug absorption and excretion, comparative pharmacology, chemotherapy, hormones, central nervous system, cardiorenal, anesthesia, toxicology, and advances in biochemical assay techniques. This volume is highly recommended for the physician in practice or research. Reviews of this type offer an excellent opportunity to keep abreast with this rapidly developing basic science.

SUMMARY. The inoculation treatment of vulvovaginitis apparently shortens the stay of patients in hospital. Whether or not it actually lessens the total duration of the disease, as compared with other methods of treatment, future investigations must determine. Old strains are more effective than fresh ones. It is desirable, though apparently not absolutely necessary, to take the index while pursuing the treatment. There is no correspondence between the index and the vaginal discharge. Our thanks are due to our colleagues on the staff of Cook County Hospital, who kindly allowed us to inoculate their patients, and to the Visiting Nurses' Association for their help in tracing patients. We also are under special obligation to Dr. Hektoen and Dr. Hamilton for much valuable help and suggestion in the opsonic work. 439 North State Street. 26 Junior Terrace. A clinical experience of ten years with institution epidemics of gonorrhea, occurring at not infrequent

Among the bacterial infections in which local antiseptic treatment does not always prove satisfactory, gonorrheal vulvovaginitis may be well included. It appears clear that whatever organisms are on the surface of the mucosa in such an infection will be disposed of, in great part at least, by the action of antiseptics, but those at a deeper level will be inaccessible to their action. Many such organisms so located may be taken care of by being washed to the surface by lymph. A great task, however, will still remain for the body to accomplish, namely: Destruction of gonococci beyond the reach of local medication through its immunizing mechanism. As the immunizing apparatus is ordinarily slow in effecting this, we attempted to call on it in a number of cases for increased labor through the stimulus of vaccine therapy, withholding at the same time all other treatment, and we present below our