Quantitative determination of FtsA at different growth rates in Escherichia coli using monoclonal antibodies. FtsA is an essential cell division protein in Escherichia coli. Its synthesis in low amounts makes the investigation of its functions difficult. Partially purified FtsA protein was obtained by solubilizing cellular inclusion bodies after overexpression of the ftsA gene for the purpose of raising monoclonal antibodies. Mice were immunized with this FtsA protein fraction and their spleen cells were fused to Sp2/0-AG14 mouse myeloma cells. Hybrid cells were screened and two clones were positively identified as FtsA monoclonal antibody producers by enzyme-linked immunosorbent assay and Western blotting. A quantitative assay using these monoclonal antibodies indicated that the average number of FtsA molecules per cell to be between 50 and 200. However, the concentration of FtsA protein normalized to total cell protein was constant over a wide range of growth rates. This finding is in agreement with the hypothesized role of FtsA protein as a stoichiometric component of the septum.


RecA protein of Escherichia coli and chromosome partitioning. Escherichia coli cells deficient in RecA protein frequently contain an abnormal number of chromosomes after completion of ongoing rounds of DNA replication. This suggests that RecA protein may be required for correct timing of initiation of DNA replication; however, we show here that initiation of DNA replication is properly timed in recA mutants. We also find that more than 10% of recA mutant cells contain no DNA. These anucleate cells appear to arise from partitioning of all the DNA into one daughter cell and no DNA into the other daughter cell. Based on these and previously published results, we propose that RecA protein is required for equal partitioning of chromosomes into the two daughter cells.


Chlamydia trachomatis Mip-like protein.

Chlamydia trachomatis Mip-like protein. A 27 kDa Chlamydia trachomatis Mip-like protein with homology of a 175-amino-acid C-terminal fragment to the surface-exposed Legionella pneumophila mip-gene product has previously been described. In this paper the entire chlamydia Mip-like sequence of C. trachomatis serovar L2 (lymphogranuloma venereum (LGV) biovar) is presented. The sequence shows high similarity to the legionella Mip protein and its C-terminal region, like that of the legionella Mip, has high amino acid similarity to eukaryotic and prokaryotic FK506-binding proteins. The chlamydial mip-like gene was detected by polymerase chain reaction (PCR) in other C. trachomatis serovars and by sequencing of the mip-like genes of serovars B and E (trachoma biovar) was shown to be highly conserved within the two major biovars of C. trachomatis. Monoclonal and polyclonal antibodies raised against the recombinant Mip-like protein failed to demonstrate surface-exposed epitopes on infectious elementary bodies or reproductive reticulate body forms either by immunofluorescence or immuno-gold electron microscopy. However, a complement-dependent inhibition of up to 91% of infectivity for cell cultures was observed with antibodies to the N-terminal fragment of the Mip-like protein suggesting that antibody-accessible epitopes are present on infectious EBs.


Partial care in mental health organizations: United States and each state, 1988. Partial care is a relatively new and rapidly-growing form of mental health care consisting of a planned program of mental health treatment services generally provided in visits of 3 or more hours to groups of patients/clients. In 1970, only one-fourth of U.S. mental health organizations provided partial care services, and patient additions to partial care programs were 56 per 100,000 civilian population. By 1988, close to half of all mental health organizations provided partial care services, and the rate of additions was more than 5 times as high. However, large increases in the number or organizations providing partial care between 1984 and 1988 are due in part to changes in definitions that are elaborated on later in the report. In 1988, multiservice mental health organizations were the most numerous providers of partial care with 1,230 of 1,310 (94 percent) providing this program. This was followed by general hospital mental health services with 332 of 1,489 (22 percent) providing partial care and private psychiatric hospitals with 236 of 447 (53 percent) providing this program. Mental health organizations providing partial care were most numerous in populous States, particularly in California (with 187 mental health organizations providing partial care), New York (174 organizations), and Pennsylvania (131 organizations). There were 212,196 patients on the rolls of partial care programs of mental health organizations in the United States (including Territories) at the beginning of 1988, and 286,715 patients were added to these programs during the year. Multiservice mental health organizations were responsible for a majority of both patients on the rolls and patient additions during the year.(ABSTRACT TRUNCATED AT 250 WORDS)


Increase in cardiac muscle fructose content in streptozotocin-induced diabetic rats. To evaluate the activation of the sorbitol pathway in cardiac muscle in diabetic rats, we measured sorbitol, fructose, and myo-inositol content in cardiac tissue obtained from control and streptozotocin-diabetic rats, with or without an 8-week insulin treatment, using gas chromatography-mass spectrometry (GC-MS). Cardiac fructose and sorbitol content in 10-week diabetic rats increased by 60-fold and 3.9-fold of those of control rats, respectively (P less than .001). In contrast, cardiac myo-inositol content in 10-week diabetic rats decreased to 56% (P less than .025) of the control value. The abnormalities in cardiac fructose, sorbitol, and myo-inositol content were completely normalized by the 8-week insulin treatment, which was initiated 2 weeks after the induction of diabetes. There was no difference in cardiac aldose reductase activity between control and diabetic rats. However, cardiac sorbitol dehydrogenase activity in diabetic rats was 151% (P less than .005) higher than that of control rats, although hepatic sorbitol dehydrogenase activity was not different between the two groups. These results indicate that the sorbitol pathway is significantly activated in cardiac tissue obtained from streptozotocin-induced diabetic rats, which results in the marked cardiac accumulation of fructose.


Effect of insulin on impaired antioxidant activities in aortic endothelial cells from diabetic rabbits. The defense system of aortic endothelial cells against oxidative stress was studied in alloxan-induced diabetic rabbits, and the effect of insulin on the antioxidant activities was estimated. Endothelial cells were prepared from 10 diabetic rabbits, 18 diabetic rabbits treated with insulin, and 10 age-matched controls after 17 days of diabetes. These cells were used for the estimation of glutathione (GSH) levels and its related enzyme activities. The antioxidant activities in these endothelial cells from diabetic rabbits were compared with those from control subjects. The concentration of GSH decreased in diabetic rabbits (1.6 +/- 0.2 nmol/mg protein [mean +/- SD] v 3.7 +/- 0.6 nmol/mg protein). Decreases in the activities of Cu, Zn-superoxide dismutase (Cu,Zn-SOD) (62.7 +/- 11.0 U/mg protein v 172.9 +/- 20.2 U/mg protein), catalase (7.6 +/- 2.1 U/mg protein v 12.3 +/- 3.2 U/mg protein), and GSH peroxidase (134.0 +/- 27.0 mU/mg protein v 179.1 +/- 26.2 mU/mg protein) were observed. The activities of other GSH-related enzymes such as GSH S-transferase or GSH reductase did not change in endothelial cells from diabetic rabbits. Most of these antioxidant activities were prevented when diabetic rabbits were treated with insulin (1 to 2 U/kg/d). These antioxidant activities were also determined in the diabetic liver and kidney. Similar decreases in the cellular defense activities and prevention of the decrease in activities by insulin were observed in the diabetic liver, while these antioxidant enzyme activities in the kidney were resistant to diabetic conditions.(ABSTRACT TRUNCATED AT 250 WORDS)


Composition of weight loss in severely obese women: a new look at old methods. Seven severely obese, outpatient dieters lost weight (mean +/- SEM, 14 +/- 1 kg), and the composition of weight lost was determined by six different models. Total body water (TBW), total body potassium (TBK), and body density, bone mineral content, and fat as determined by dual photon absorptiometry (DPA) were measured while subjects were weight-stable, before and after weight loss. Fat loss was calculated by three two-compartment models (2C-TBW, 2C-TBK, and hydrodensitometry [2C-HD]), one three-compartment model (HD with correction for water content of fat-free mass [FFM], 3C), and one four-compartment model (HD with correction for water and mineral content of FFM, 4C), and was measured directly by DPA. Mean composition of weight loss was similar for all models (mean weight lost as fat: 89% for DPA, 91.5% for 4C, 89% for 3C, 88.6% for 2C-HD, and 87% for 2C-TBW) except 2C-TBK (weight lost as fat, 66%). There was a much wider range of individual values for the 2C-TBW and 2C-TBK models (17% to 138% and 18% to 93%, respectively) than for the multicompartment models (63% to 112%) and DPA (76% to 107%). Almost opposite results were obtained for the same individual when using the 2C-TBK and 2C-TBW models. The discrepancy between these models was due to the inverse relationship between changes in TBW and TBK in the group as a whole (r = -.34, NS). In addition, TBK loss was found to be dependent on the initial level of hyperinsulinemia, calculated as the area under the 2-hour oral glucose tolerance curve.(ABSTRACT TRUNCATED AT 250 WORDS)


Manganese + 2 exhibits dynamic binding to multiple ligands in human plasma. Plasma from fasted adult male subjects was labeled in vitro with 54MnCl2 and then fractionated using several techniques. Molecular sieve chromatography showed that the major 54Mn-containing peak had a very low molecular weight (VLMW), although four other significant peaks, one of which corresponded to the mass of transferrin (Tf), were also observed. The 54Mn content of the Tf peak increased with increasing incubation time in vitro, suggesting the oxidation of Mn+2 to Mn+3 before its association with Tf. This time-dependent effect was verified using affinity chromatography consisting of immobilized anti-Tf. Electrophoretic analyses of plasma yielded equivocal results, indicating a limited value of this method for investigating plasma manganese localization. The above findings are discussed in the context of factors that influence the oxidation and metabolism of Mn2+ in human plasma.


Treatment of primary chylomicronemia due to familial hypertriglyceridemia by omega-3 fatty acids. Primary familial forms of chylomicronemia can lead to acute life-threatening complications, especially acute pancreatitis. The main aim of therapy is to avoid this so-called chylomicronemia syndrome. In 12 patients with primary chylomicronemia due to familial hypertriglyceridemia, the addition of 2.16 g omega-3 fatty acids over 4 weeks and 4.32 g for 8 weeks resulted in a decrease of serum triglyceride levels from 1,624 +/- 333 to 894 +/- 241 mg/dL after 12 weeks. Cholesterol and triglyceride levels in the chylomicron fraction were reduced concomitantly, the apolipoprotein B-100/B-48 ratio increased, very--low-density lipoprotein (VLDL) triglycerides, VLDL cholesterol, and total cholesterol levels decreased, and low-density lipoprotein (LDL) cholesterol showed a tendency to increase, but this finding did not reach significance. High-density lipoprotein (HDL) cholesterol levels remained unchanged, as did the levels of apolipoproteins A-I, A-II, and E, and lipoprotein(a). Apolipoprotein B levels decreased significantly. The decrease of triglyceride levels to still-elevated concentrations was accompanied by a substantial decrease in plasma and whole-blood viscosity and erythrocyte aggregation, which reached normal values. As in chylomicronemia, complications usually occur at triglyceride levels higher than 1,500 mg/dL; patients can still profit from treatment with omega-3 fatty acids, even though triglyceride levels are still substantially elevated. No clinically relevant side effects occurred, with the exception of the manifestation of diabetes mellitus in one patient, which could be reversed after discontinuation of treatment.


Effect of short-term fasting on nocturnal melatonin secretion in obesity. To investigate whether the function of pinealocytes is altered in obesity, nocturnal melatonin (MT) secretion was determined in nine healthy subjects and compared with that of eight obese individuals. Serum MT levels were measured every second hour between 6:00 PM and 8:00 AM, and total nocturnal MT secretion (as reflected by the MT incremental area), MT peak time, and nocturnal urinary MT excretion were determined. None of these parameters differed significantly in the two groups. The obese subjects were reinvestigated after 2 days of complete fasting. This caused a decrease in body weight and basal blood glucose levels of 2.6 +/- 0.2 kg (mean +/- SEM, P less than .001) and 1.5 +/- 0.2 mmol/L (P less than .001), respectively, whereas serum cortisol levels remained unchanged. Short-term fasting reduced nocturnal MT secretion, as evidenced by MT incremental areas, which were reduced from 2.01 +/- 0.26 before fasting to 1.64 +/- 0.26 nmol/L.h after fasting (P less than .02). MT secretion peaks were reached simultaneously, and urinary MT excretion values did not change significantly in fasting. To see whether glucose supplementation during short-term fasting would normalize nocturnal MT secretion, we gave an additional seven obese subjects eight small oral doses of glucose (each dose, 0.5 g/kg body weight) at regular intervals during a 2-day fast. Their body weight decreased by 2.8 +/- 0.4 kg (P less than .001), but blood glucose and cortisol concentrations were similar before and after the glucose-supplemented fast, as was the nocturnal MT secretion.(ABSTRACT TRUNCATED AT 250 WORDS)


Role of impaired intracellular glucose metabolism in the insulin resistance of aging. The insulin resistance of aging is characterized by both reduced glucose uptake and impaired intracellular glucose metabolism. The aim of this study was to determine whether impaired intracellular glucose metabolism contributes to insulin resistance in the elderly independent of reduced glucose uptake. To address this question, glucose uptake in non-obese elderly males was matched to controls using the glucose clamp technique, and intracellular glucose metabolism was assessed in vivo by indirect calorimetry and in vitro by skeletal muscle biopsy for glycogen synthase activity. When elderly subjects were compared with controls at an equivalent basal glucose uptake of approximately 2.5 mg/kg fat-free mass (FFM)/min, muscle glycogen synthase activity was similar (fractional velocity of glycogen synthase at 0.1 mmol/L glucose-6-phosphate [FV0.1], 0.06 +/- 0.1 and 0.07 +/- 0.1), but whole-body rates of glucose oxidation were reduced (1.36 +/- 0.12 v 1.90 +/- 0.11 mg/kg FFM/min, P less than .05). During 40-mU/m2/min hyperinsulinemic clamps at matched rates of glucose uptake (approximately 10.7 mg/kg FFM/min in both groups), glycogen synthase activity was again similar (FV0.1, 0.15 +/- 0.02 and 0.14 +/- 0.02), and glucose oxidation remained reduced in the elderly (4.18 +/- 0.25 v 4.77 +/- 0.17 mg/kg FFM/min, P less than .05). Only during clamps in the maximal range of glucose uptake (approximately 29.5 mg/kg FFM/min) was glucose oxidation between the groups comparable (5.97 +/- 0.50 and 5.75 +/- 0.31 mg/kg FFM/min). Plasma free fatty acid (FFA) concentrations, fat oxidation, and protein oxidation were similar under all study conditions.(ABSTRACT TRUNCATED AT 250 WORDS)


Low and marginal copper intake by postweanling rats: effects on copper status and resistance to carbon tetrachloride hepatotoxicity. Copper feeding studies in rats are generally initiated at weaning. This study examined whether a 6-week feeding of low or marginal Cu levels (0.2 or 2.5 ppm) to rats initially weighing 135 g produced deleterious effects. Controls were fed 8 ppm Cu. Liver Cu-Zn superoxide dismutase activities paralleled Cu intake. Plasma ceruloplasmin activities were very low for both low and marginal Cu consumption. Low but not marginal Cu intake caused a low body weight, high plasma cholesterol level, anemia, cardiac hypertrophy, and a high degree of hepatic plasma membrane injury 24 hours after CCl4 injection (150 microL/kg intraperitoneally [IP]). In summary, low and marginal Cu intakes produced low Cu enzyme activities, while low Cu intake produced pathological symptoms and poor resistance to an oxidative stress.


Genetically obese rats with (SHR/N-cp) and without diabetes (LA/N-cp) share abnormal islet responses to glucose. To assess the effect of hyperglycemia on the function of islets obtained from obese rats, the behavior of isolated islets from LA/N-corpulent (nondiabetic obese) and SHR/N-corpulent (diabetic obese) male rats was examined and compared. Islets from both genetic models showed a left-shifted glucose dose-response curve for insulin release (concentrations for half-maximal release, 5 to 6 mmol/L v 12 to 13 mmol/L in LA/N lean littermates and 3 mmol/L v 10 mmol/L in lean SHR/N). When insulin release was expressed per unit islet volume, the fourfold to fivefold enlarged islets from both obese diabetic and obese nondiabetic rats showed decreased insulin secretory response in high (16.5 to 28 mmol/L) glucose concentrations, although the decrease was more severe in the diabetic rats. Glucose-stimulated insulin release by islets from both models was relatively resistant to inhibition by 1.2 mmol/L mannoheptulose (eg, 82% +/- 3% inhibition in LA/N lean v 16% +/- 8% in LA/N obese), although nearly complete inhibition was observed with 16 mmol/L mannoheptulose (96% v 85%, NS). Islets of obese diabetic rats were also resistant to the calcium-channel blocker, verapamil, suggesting an abnormal pathway of stimulus-secretion coupling for glucose. Glucose oxidation to carbon dioxide was increased in both obese models at all glucose concentrations when expressed per islet. In data expressed per unit volume, the larger islets from the obese-nondiabetic rats showed a left-shifted dose-response curve with an unchanged maximum rate of glucose oxidation at high (16.5 mmol/L) glucose concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)


The influence of genetic background on the expression of mutations at the diabetes (db) locus in the mouse. VI: Hepatic malic enzyme activity is associated with diabetes severity. The severity of diabetes produced by the mutation diabetes (db) in the mouse is markedly strain-dependent. When the db mutation is maintained in the C57BL/KsJ (BKs) and DBA/2J strains, severe diabetes is observed, whereas when it is maintained in the C57BL/J (B6) inbred background, a mild, well-compensated diabetes is observed. Our studies on the regulation of malic enzyme activity showed that both the BKs and DBA/2J strains carried the b allele at the malic enzyme regulatory (Modlr) locus and had low enzyme activity, while the B6 strains carried the a allele and had malic enzyme activity two to three times that seen in the BKs and DBA/2J strains. To assess any role of malic enzyme activity in modulating diabetes severity, we produced an F2 generation of diabetic mice using BKs-db/+ and B6-db/+ mice as progenitors. Male diabetic mice of the F2 generation segregated into three groups with respect to diabetes severity. The concordance observed between diabetes severity and malic enzyme activity seen in these three groups suggests a major role for a gene (or a closely linked gene) regulating malic enzyme activity to be responsible for much of the genetic background effects observed with the db mutation in the mouse.


Metabolic response to cottage cheese or egg white protein, with or without glucose, in type II diabetic subjects. Test meals with 25 g protein in the form of cottage cheese or egg white were given with or without 50 g glucose to male subjects with mild to moderately severe, untreated, type II diabetes. Water was given as a control meal. The glucose, insulin, C-peptide, alpha amino nitrogen (AAN), glucagon, plasma urea nitrogen (PUN), nonesterified fatty acid (NEFA), and triglyceride area responses were determined using the water meal as a baseline. The glucose area responses following ingestion of cottage cheese or egg white were very small compared with those of the glucose meal, and were not significantly different from one another. The serum insulin area response was 3.6-fold greater following ingestion of cottage cheese compared with egg white (309 v 86 pmol/L.h). The simultaneous ingestion of glucose with cottage cheese or egg white protein decreased the glucose area response to glucose by 11% and 20%, respectively. When either protein was ingested with glucose, the insulin area response was greater than the sum of the individual responses, indicating a synergistic effect (glucose alone, 732 pmol/L.h; glucose with cottage cheese, 1,637 pmol/L.h; glucose with egg white, 1,213 pmol/L.h). The C-peptide area response was similar to the insulin area response. The AAN area response was approximately twofold greater following ingestion of cottage cheese compared with egg white. Following ingestion of glucose, it was negative. When protein was ingested with glucose, the AAN area responses were additive. The glucagon area response was similar following ingestion of cottage cheese or egg white protein. Following glucose ingestion, the glucagon area response was negative.(ABSTRACT TRUNCATED AT 250 WORDS)


Epinephrine directly antagonizes insulin-mediated activation of glucose uptake and inhibition of free fatty acid release in forearm tissues. To determine whether the anti-insulin effect of epinephrine is due to a direct antagonism on target tissues or is mediated by indirect mechanisms (systemic substrate and/or hormone changes), insulin and epinephrine were infused intrabrachially in five normal volunteers using the forearm perfusion technique. Insulin (2.5 mU/min) was infused alone for 90 minutes and in combination with epinephrine (25 ng/min) for an additional 90 minutes, so as to increase the local concentrations of these hormones to physiological levels (60 to 75 microU/mL and 200 to 250 pg/mL for insulin and epinephrine, respectively). Systemic plasma glucose and free fatty acids (FFA) concentrations remained stable at their basal values during local hormone infusion. Forearm glucose uptake (FGU) increased in response to insulin alone from 0.8 +/- 0.2 mg.L-1.min-1 to 4.3 +/- 0.8. Addition of epinephrine completely abolished the insulin effect on FGU, which returned to its preinfusion value (0.7 +/- 0.2). Forearm lactate release was slightly increased by insulin alone, but rose markedly on addition of epinephrine (from 5.2 +/- 0.8 mumol.L-1.min-1 to 17 +/- 2; P less than .02). During infusion of insulin alone, forearm FFA release (FFR) decreased significantly from the postabsorptive value of 1.76 +/- 0.25 mumol.L-1.min-1 to 1.05 +/- 0.11 (P less than .01). Epinephrine addition reverted insulin suppression of FFR, which returned to values slightly above baseline (2.06 +/- 0.47 mumol.L-1.min-1; P less than .05 v insulin alone). The data demonstrate that epinephrine is able to antagonize directly insulin action on forearm tissues with respect to both stimulation of glucose uptake and inhibition of FFA mobilization.


Modeling DNA structures: molecular mechanics and molecular dynamics. Model building studies may be used to supplement structurally low resolution experimental data with detailed three-dimensional hypothetical atomic models. Because of the strong relation between structure and function in biological molecules such models may give a consistent, integral view of a wealth of experimental data. In most cases such models will predict the outcome of certain experiments. The outcome of these experiments will often either confirm the model may be used for further refinement or even demand a major revision of the model. Coordinates obtained from X-ray fiber diffraction data or in special cases single-crystal data may provide the elements for DNA or RNA model building. Local and nonlocal optimization may be used to refine these structures and to evaluate their statistical significance as estimated by a chosen force field. Appreciable progress using nonlocal optimization procedures can only be expected if the dimensionality of the problem can be reduced sufficiently to the relevant degrees of freedom. Taking advantage of structural symmetries may critically improve the convergence while refining the target molecule or its building blocks. Monte Carlo and molecular dynamics methods allow one to calculate averaged quantities. In addition, molecular dynamics provides time evolutions of certain averages. During the simulation of certain physical properties of molecules a huge amount of data will be generated. They will provide many answers, but these answers may not always apply to the original question. So what type of questions will be reliably answered by a force field? Relatively safe answers concern the local geometry of the molecules. If a conformation leads to strong distortions of bond distances or angles or to close van der Waals contacts, this conformation can safely be rejected. Optimizing such unfavorable structures energetically may lead to structures showing how to avoid such distortions. More difficult are energetical questions: which of two conformers is more stable, or what is the free energy of the substrate in the active site? One cannot always be sure that the force field provides the correct answer. Therefore, one should pose only those questions which can be checked experimentally. Because of the many possible answers, the experiment may benefit by starting with a choice proposed by the simulation. The application of this procedure to curved DNA and the DNA four-way junction was successful.


Solution behavior of DNA studied with magnetically induced birefringence. Magnetically induced birefringence can provide useful information on the physical properties of nucleic acids in a variety of forms. The technique is particularly sensitive to structural changes. Difficulties arise with drawing precise structural conclusions principally because of doubts about the relative contributions of the form and intrinsic components to the optical anisotropy. Nevertheless results so far suggest that this might not be a severe limitation. The induced birefringence is very sensitive to cooperative behavior; consequently the transition from an isotropic to a liquid crystalline phase can be followed in detail. Finally the high degree of magnetic orientation often achieved with liquid crystals assists their study with other physical techniques.


Automated online transition from the medical record to the psychiatric literature. Psych Topix is a knowledge-based program which guides the clinician from an on-line clinical report to a search of the psychiatric literature or of other relevant databases. It provides this guidance by using an outline of key topics in a clinical field to provide "concept-based" links. Each topic is augmented with an activation expression to signal when that topic is potentially relevant to a case, and with database search expressions to allow focused retrieval of information. The bibliographic retrieval component of Psych Topix is currently operational as part of the daily, routine operation of a psychiatric consultation service. The system is also implemented in a demonstration mode to provide retrieval from three additional textual databases. The current Psych Topix system provides a working demonstration of the clinical feasibility of using concept-based links to facilitate the focused, automated transition between on-line medical databases.


Validation of the AI/RHEUM knowledge base with data from consecutive rheumatological outpatients. The diagnostic accuracy of AI/RHEUM, an experimental expert system for support in the diagnosis of rheumatic diseases, was assessed using a collection of data in a cohort of 1,570 consecutive outpatients of a Dutch rheumatological clinic. Computer diagnoses based on these data and diagnostic predictions made by rheumatologists were compared with reference diagnoses that had been obtained by consensus of rheumatologists after 6-12 months follow-up. Performance of the tested version of the AI/RHEUM knowledge base is presented by various methods. Sensitivity varied between 29% and 100% for different rheumatological diseases. Average sensitivity and specificity for all 26 diagnoses present in the knowledge base were 67% and 98%, respectively. Performance according to the level of confidence indicated that 78% of the "definite", 65% of the "probable", and 33% of the "possible" conclusions made by AI/RHEUM were in agreement with the reference diagnoses. These results approximated the predictions made by rheumatologists after a single, initial examination. The system was less accurate than it had appeared in previous evaluation studies with complex clinical cases. The AI/RHEUM knowledge base needed refining to diagnose early rheumatic complaints. This study further illustrates the need for objective and informative parameters for expressing accuracy of diagnostic support systems.


Towards an essential data set: applicability in the domain of maternal health services. There is a need for consensus on the quantity of data that must be available in a computer-based information system of a health care organization. In this paper we take up the issue of defining the data content of an information system and introduce the concept of Essential Data Sets with an explicit methodology which was applied to define a data set for the Maternal Health Services program. A key step in the method was a recognized technique used in systems development process called data modelling, in this case infological modelling, by an interdisciplinary group. A preliminary set of 86 data elements was identified and it provided the foundation for development of an application software for discussion and a real-world testing framework. The acceptability of the data set was tested in a laboratory perspective by retrospective data entry from records of 94 pregnant women registered at a maternal health care center in Sweden. Data from a total of 1,318 prenatal visits, an outcome visit, and a postnatal visit for each woman was entered into a computer using the software, with no loss of information. Thus, in a short-term perspective the acceptability of the data set was demonstrated. The software has since been implemented for pilot prospective studies at sites in India and Sweden. The use of a common data protocol is an essential foundation for patient outcome research, especially as the trend of health care management has changed from a "process of care" orientation to an "outcome of care" orientation.


An intelligent computer-assisted instruction system designed for rural health workers in developing countries. This paper describes an intelligent computer-assisted instruction system that was designed for rural health workers in developing countries. This system, called Consult-EAO, includes an expert module and a coaching module. The expert module, which is derived from the knowledge-based decision support system Tropicaid, covers most of medical practice in developing countries. It allows for the creation of outpatient simulations without the help of a teacher. The student may practice his knowledge by solving problems with these simulations. The system gives some initial facts and controls the simulation during the session by guiding the student toward the most efficient decisions. All student answers are analyzed and, if necessary, criticized. The messages are adapted to the situation due to the pedagogical rules of the coaching module. This system runs on PC-compatible computer.


Dilemmas at a primary health care center: a baseline study for computer-supported cooperative health care work. For the development of computer-supported cooperative health care work this study investigated, based upon activity theory, daily dilemmas encountered by the members of interprofessional primary health care work groups. The entire staff at four Swedish primary health care centers were surveyed, 199 personal interviews being conducted by the Critical Incident Technique. Medical dilemmas were mainly reported by general practitioners and nurses, organizational dilemmas by laboratory staff, nurses' aides, and secretaries, and dilemmas in the patient-provider relation by nurses, nurses' aides, and secretaries. Organizational and communication dilemmas reported by nurses, nurses' aides, and secretaries often had their cause outside the control of the individual professional. These dilemmas were often "caused" by other group members (general practitioners or nurses), e.g., by not keeping appointment times or by not sharing information with patients. The implication for computer-supported cooperative health care work is that computer support should be planned on two levels. Collective work activity as a whole should benefit from individual clinical decision support for general practitioners and nurses. However, since most patient communication and organizational problems occurred at group level, group process support is required in these areas.


Application of adjusted survival curves to renal transplant data. An important means in the analysis of survival time data is the estimation and graphical representation of survival probabilities. In this paper unifactorial parametric and non-parametric survival curve estimators and two types of adjusted survival curves based on a parametric multifactorial approach are applied to renal transplant data. It is shown that the resulting survival curves can differ substantially. The unifactorial survival curves yield biased results in case of serious disequilibrium in the data. This drawback of the unifactorial methods has been overcome by the use of adjusted survival curves which take possible distortions in the data set into account. The benefits of adjusted survival curves in assessing potentially prognostic factors are elucidated by the application to data from renal transplantation.


Maximum likelihood estimation and testing of a poisson regression model. A Poisson regression model is proposed for the analysis of incidence rates presented in a two-way table classified by two categorical variables. It is shown that the likelihood function is the same as that using Glasser's exponential covariate model. An algorithm is given to solve the maximum likelihood estimates of the regression parameters. The model is evaluated via deviance and the method is illustrated with an example. Some extensions of the model are discussed.


Adhesion of bacteria on pectin casted films. Pectin and non-pectin degrading bacteria were tested for their ability to adhere to a film casted of low methoxylated pectin (polygalacturonic acid). Klebsiella oxytoca and a newly isolated strain of Escherichia coli adhered to the film, whereas only K. oxytoca was able to utilize pectin as a sole carbon source. Other E. coli strains, containing plasmids with pectinolytic enzymes, did not adhere to the solid pectin film. Therefore, the ability of the bacteria to adhere to the films was not correlated with their ability to degrade pectin. When the solubilization (dissolution) of pectin matrices (tablets) was analysed with and without K. oxytoca, a significant retardation in the dissolution rate was observed in the presence of K. oxytoca, suggesting the formation of a biofilm on the matrix or sedimentation of insoluble pectin salts.


Immunomodulating activities of Corynebacterium xerosis cell-wall fractions. Corynebacterium xerosis cell-wall fractions were studied by electron microscopy and analysed for immunomodulating activity. Dramatic splenomegaly occurred following the injection of whole cells or a purified cell-wall fraction (PF), but not with a further purified peptidoglycan (PEP) fraction. Both PF and PEP acted as B-cell mitogens and had adjuvant capabilities comparable to commercial adjuvants. Only the PF fraction enhanced peritoneal natural killer cell (NK) activity, paralleling the splenomegaly response. When spleens from mice injected with PF or PEP were analysed for their abilities to respond to mitogens and for the presence of suppressor cells, reduced mitogenic responses occurred only in PF-injected mice during the peak of splenomegaly. Spleens from both PF- and PEP-injected mice contained suppressor cell activity which peaked 2 weeks post-injection. This activity was primarily directed at B-cell responses to lipopolysaccharide (LPS). C. xerosis cell-wall fractions thus offer great potential as a new immunomodulator.


Origins of BCG surface charge: effect of ionic strength and chemical modifications on zeta potential of Mycobacterium bovis BCG, Tice substrain, cells. The zeta potential of washed Tice substrain BCG organisms was measured over a range of ionic strengths from I = 0.005 to 0.1 M. No change in the isoelectric point of 3.4-3.7 was evident. Proteolytic enzymes (trypsin/chymotrypsin, pepsin, papain and pronase) and fluorodinitrobenzene abolished the cationic charge, suggesting that this is substantially due to amino groups associated with protein. Neither hot HCI nor cold trichloroacetic acid affected the charge, indicating that ionic groups are not associated with extractable polysaccharides. Methanolysis, treatment with HF and carbodiimide, and cationic detergent (cetyltrimethylammonium bromide) binding indicated that the negative charge was provided by carboxylic acids, phosphoesters and strong acidic groups, possibly sulphates. Standardless quantitative X-ray microanalysis revealed the presence of phosphorus and sulphur on the surface of actively growing BCG colonies.


Isolation of an Enterobacter agglomerans strain with inhibitory activity on Pseudomonas aeruginosa. Antagonism between Enterobacter agglomerans and Pseudomonas aeruginosa was due to an extracellular substance produced by E. agglomerans which accumulates in the culture medium. This substance was also toxic to Escherichia coli and Staphylococcus albus and its MIC for P. aeruginosa ranged between 7.8 and 3.9 micrograms/ml. However, when the extracts from the culture medium were irradiated from 2 h with white light (15 mW/cm2), the MIC was lower (3.9-1.9 micrograms/ml) suggesting that the active substance was a phototoxin.


Mechanism of action of chlorhexidine diacetate and phenoxyethanol singly and in combination against gram-negative bacteria. Chlorhexidine diacetate and the aromatic alcohol, phenoxyethanol in combination had an enhanced bacteriostatic action against Escherichia coli and Pseudomonas aeruginosa strains. Investigations of potassium (K+) ion leakage by means of a potassium electrode and a radioactive method, employing 86Rb, indicated that the combination accelerated the rate of leakage from the cell. Leakage of pentose was also found to be enhanced in the presence of the combination compared with either drug alone.


Quantitative determination of Escherichia coli from coliforms and faecal coliforms in sea water. Escherichia coli concentration in sea water was determined by the MUG test after primary growth on membrane filters used to determine total coliforms or faecal coliforms. A good correlation (r = 0.86) was found between E. coli obtained from coliforms versus those from faecal coliforms. Verification procedures showed that all the MUG-positive colonies obtained on both media were E. coli. Evaluation of this data and the literature indicated that this technique for estimation of E. coli in sea water is a useful addition to laboratory procedures without generally increasing the time and the expense of the analysis of recreational water.


Photosensitizing activity of water- and lipid-soluble phthalocyanines on prokaryotic and eukaryotic microbial cells. The photosensitizing activity of lipophilic zinc-phthalocyanine (Zn-Pc) and its water-soluble sulphonated derivative (Zn-PcS) towards Streptococcus faecium and Candida albicans was studied and correlated with the amount of cell-bound photosensitizer. With both micro-organisms Zn-PcS was more tightly bound in larger amounts than Zn-Pc in the protoplasts of the cytoplasmic membrane. As a consequence, the photoinduced damage in S. faecium initially involved membrane proteins, while DNA was modified only upon prolonged irradiation. For C. albicans only Zn-PcS showed a preferential affinity for the spheroplasts and the decrease in cell survival was not accompanied by detectable modifications of the electrophoretic pattern of membrane proteins. The photoinduced ultrastructural alteration of both micro-organisms suggests damage at membrane level. This would indicate the involvement of different targets in bacteria and yeast for phthalocyanine photosensitization.


Cell surface saccharide differences in drug-susceptible and drug-resistant strains of Trichomonas vaginalis. Surface carbohydrates of drug-resistant and drug-susceptible strains of Trichomonas vaginalis were analysed using lectins. The presence of D-GalNAc, D-Gal and mannose-like residues was detected in T. vaginalis. Marked differences in exposed surface carbohydrates were documented, e.g. wheat germ agglutinin (WGA) selectively agglutinated the drug-susceptible strain whereas drug-resistant parasites reacted preferentially with concanavalin A (Con A). In drug-resistant, but not in drug-susceptible strains, trypsinization induced the appearance of soybean agglutinin. Binding studies using fluorescein-labelled WGA and Con A essentially confirmed the agglutination experiments. Both the intense cell agglutination and the fluorescent WGA-binding displayed by a drug-susceptible strain, were completely nullified by neuraminidase treatment, suggesting the presence of an exposed sialic acid moiety on the T. vaginalis surface.


Structure activity relationships of stendomycin, a lipopeptide antibiotic from Streptomyces. A strain of Streptomyces which produced stendomycin, a lipopeptide antibiotic, was grown in culture media containing various amino acids as nitrogen substrates. The nature of the fatty acid component of stendomycin was dependent on the nature of the amino acid present in the medium, but this did not affect antibiotic activity. Modifications in the peptide moiety resulted in a loss of antifungal activity.


Molecular cloning and nucleotide sequencing of a novel aminoglycoside 6'-N-acetyltransferase gene from an R-plasmid of Salmonella typhimurium S24 isolated in Taiwan. A conjugative aminoglycoside resistance plasmid pST2 has been isolated from Escherichia coli K-12 14R525, which was mated with a clinical isolate of Salmonella typhimurium S24. A novel resistance gene of aminoglycoside 6'-N-acetyltransferase[AAC(6')] was cloned from plasmid pST2 on a 1,393 kilobase (kb) of SphI-SalI fragment into vector pACYC184 and pUC18. This novel AAC(6') gene in plasmid pST2 acetylated kanamycin, amikacin, dibekacin, tobramycin, gentamicin, netilmicin, and sisomicin. The complete nucleotide sequence of the novel AAC(6') gene and its neighboring sequences were also determined. Minicell experiments detected only one protein of 24.7 kilodaltons (kDa) translated from an open reading frame of the 618 base pairs (bp) gene.


Bacteriostatic and bactericidal activity of antituberculosis drugs against Mycobacterium tuberculosis, Mycobacterium avium-Mycobacterium intracellulare complex and Mycobacterium kansasii in different growth phases. Bacteriostatic and bactericidal activities of rifampicin, isoniazid, streptomycin, enviomycin and ethambutol against Mycobacterium tuberculosis, Mycobacterium avium--M. intracellulare complex and Mycobacterium kansasii were studied in different growth phases. Bacteriostatic activities of the drugs were similar in different growth phases, except isoniazid. M. tuberculosis was much less susceptible to isoniazid in the lag phase than in the log and the stationary phases. In contrast, bactericidal activity was influenced by the growth phase. M. tuberculosis was killed by isoniazid, streptomycin and rifampicin. The bactericidal activity of isoniazid was strongest. The bactericidal activity of isoniazid and streptomycin was most marked in the log phase. M. avium complex and M. kansasii resisted the bactericidal activity, but some strains of M. avium complex were killed by streptomycin and enviomycin, and the activities of these two drugs were most marked in the lag phase.


Bactericidal activities of rat defensins and synthetic rabbit defensins on Staphylococci, Klebsiella pneumoniae (Chedid, 277, and 8N3), Pseudomonas aeruginosa (mucoid and nonmucoid strains), Salmonella typhimurium (Ra, Rc, Rd, and Re of LPS mutants) and Escherichia coli. Rat defensins were purified and tested for in vitro bactericidal assay against gram-positive and gram-negative bacteria. Staphylococcus aureus (209P, Cowan I, Smith diffuse and Smith compact) were resistant to defensins, whereas Staphylococcus epidermidis, Staphylococcus saprophyticus, Micrococcus lysodeikticus and Bacillus subtilis were less sensitive. Gram-negative bacteria, such as Pseudomonas aeruginosa (mucoid and K) and Klebsiella pneumoniae (Chedid, 277, and 8N3 which were heavily capsulated, moderately capsulated and noncapsulated, respectively) were all very sensitive to defensins and killed within 20 min. Escherichia coli was moderately sensitive and the rough mutants of lipopolysaccharide (LPS) of Salmonella typhimurium LT2, such as Ra, Rc, Rd, and Re were equally sensitive to defensins, being killed within 40 min. Lysozyme did not show any bactericidal activity except against M. lysodeikticus and B. subtilis, whereas it enhanced the bactericidal activity of defensins against P. aeruginosa, E. coli, and K. pneumoniae and suppressed the killing activity of defensins against S. typhimurium and S. aureus. With regard to the three synthetic rabbit defensins, NP1, NP4, and NP5, NP1 showed strong bactericidal activity against K. pneumoniae 277, comparable to that of rat defensins. Neither NP4 nor NP5 showed any bactericidal activity, while NP5 rather enhanced the bactericidal activity of NP1 against K. pneumoniae 277.


Detection of leishmania antigen in kala azar patients using monoclonal antibodies. Twenty-one monoclonal antibodies were produced against promastigote antigens of Leishmania donovani. Five monoclonal antibodies (Hyb.17, 6, 5, 4 and 2) identifying molecules associated with various L. donovani antigenic determinants ranging from 42-116 kDa were selected as 'capture antibodies' and compared with specific anti-leishmania antisera for detection of circulating leishmania antigens in kala azar patients' sera in a competitive-enzyme-linked immunosorbent assay system (ELISA). The anti-leishmania antisera could detect circulating antigen in 30% of kala azar cases while out of the five monoclonals, Hyb.17 could effectively detect circulating leishmania antigen in 85.4%. The efficacy of Hyb.6 was however low (31.7%). The antigens recognized by these monoclonal antibodies in the western blot assay could possibly represent the ones circulating in sera of patients suffering from kala azar. A cocktail of these monoclonal antibodies may be more useful than the conventional polyclonal antisera in detection of circulating antigen for clinical diagnosis of kala azar.


Establishment of hybridoma cells with natural killer(NK)-like activity against syngenic tumor cells. The 4D1D4 hybridoma cells were derived from the fusion of spleen cells from BALB/c nude mice with NS-1 mouse myeloma cells. The surface phenotypes of 4D1D4 hybridoma cells were Thy-1.2+, L3T4 (CD4)-, Lyt-2 (CD8)-, Asialo GM1+ and p-55 interleukin-2 (IL-2) receptor (CD25)-. This phenotypic pattern was consistent with the surface phenotype of NK cells. The 4D1D4 cells showed the definite killer activity against a syngenic tumor cell line, RL male-1, but not against an allogenic YAC-1 line. The killer activity of the 4D1D4 cells was not affected by the addition of exogenous IL-2. It was, therefore, suggested that 4D1D4 cells might be representative of resting NK cells with expression of no functional IL-2 receptors. The hybridoma technology might be useful for establishment of the cloned NK cells.


A simple purification method of Vibrio cholerae non-O1 hemagglutinin/protease by immunoaffinity column chromatography using a monoclonal antibody. A new simple purification method (I) for Vibrio cholerae non-O1 hemagglutinin/protease (NAG-HA/P) was developed. The method (I) requires only an immunoaffinity column chromatography using a monoclonal antibody against NAG-HA/P. The method (I) is much simpler than previously reported purification method (II) (Honda, T. et al, Infection and Immunity 57: 2799-2803, 1989) which required four or more complicated chromatographic procedures. Method (I) also gave an improved recovery rate (about 27%) compared with (II). The molecular weight of NAG-HA/P purified by method (I) was mainly 34 kilodaltons (kDa) with a little of 32 kDa, whereas that of NAG-HA/P purified by (II) was usually 32 kDa. Immunological analysis by the Ouchterlony double gel diffusion test and Western blotting test using polyclonal antibody against 32 kDa protein revealed that the 34 and 32 kDa proteins are immunologically indistinguishable and thus it is supposed that 34 K protein is an isoform or a preform of the 32 K protein.


Oligo-2',5'-adenylate synthetase activity in cells persistently infected with human T-lymphotropic virus type I (HTLV-I). Spontaneous production of interferon-gamma (IFN-gamma) was shown in several T-lymphoblastoid cell lines persistently infected with human T-lymphotropic virus (HTLV-1). However, the produced IFN-gamma was not always associated with the induction of the antivirus state. The induction of oligo-2',5'-adenylate synthetase (2-5AS) by IFN was studied in five human T-cell lines persistently infected with HTLV-I (MT-1, MT-2, SMT-1, HUT 102 and OKM-2). Four cell lines are able to produce IFN-gamma spontaneously, while the OKM-2 cell line is not. Poor induction of 2-5AS was recognized in three (MT-1, MT-2 and SMT-1) of the four cell lines producing IFN-gamma, though the poor induction was improved after long-term cultivation of cells with IFN-alpha. On the contrary, in the OKM-2 cell line, significant activity of the enzyme was induced by IFN-alpha. Induction of 2-5AS was not correlated with cell growth inhibition, but with the antivirus state. Furthermore, an inverse relationship between IFN-gamma production and 2-5AS induction was demonstrated in these cell lines with the exception of HUT 102 cells.


Proposal of a new scheme for the serological typing of Enterococcus faecalis strains. No systematic study on serotyping of Enterococcus faecalis has been reported since 1964 when M.E. Sharpe conducted serotyping of group D streptococcus in U.K. So, we attempted to re-evaluate serotyping of E. faecalis. For this purpose, we received 42 Sharpe's strains and first examined for their biochemical characteristics as E. faecalis. Only 9 of the 42 strains were identified as E. faecalis. We raised rabbit antisera against a large number of E. faecalis strains, including the 9 Sharpe's strains, 2 strains obtained from CDC in U.S.A. and 36 strains isolated from patients hospitalized in different cities of Japan. From the results of cross-agglutination tests and absorption tests performed on these antisera using a large number of E. faecalis strains, we were able to classify 21 distinct serotype strains and to prepare 21 monospecific typing antisera by absorption of the antisera to the type strains with appropriate cross-agglutinating strains. When 832 E. faecalis strains were serotyped with the 21 typing antisera, 638 strains (76.7%) were typable. Thus, we propose a provisional scheme of 21 distinct serovars in E. faecalis.


Impact of Bifidobacterium longum on human fecal microflora. The effects of Bifidobacterium longum feedings for five weeks on the fecal microflora, water contents, pH values, ammonia concentration, and beta-glucuronidase activity were investigated in five healthy human volunteers. Although numbers of major bacterial groups of the fecal microflora were not changed by the bifidobacteria feedings, a remarkably decreasing number of lecithinase-negative clostridia was observed. The percentage of lecithinase-negative clostridia and bacteroides to the total bacterial numbers isolated were decreased during the feedings and numbers of C. paraputrificum and C. innocuum were reduced. A significant reduction of fecal pH values for the last week of the feeding was observed. Ammonia concentration and beta-glucuronidase activity in the feces during the feedings were significantly lower than those before or after the feedings. The oral supplement of B. longum may be introduced to improve the fecal properties such as fecal ammonia concentration and beta-glucuronidase activity, but not the composition of fecal flora.


Cloning and nucleotide sequence analysis of a chloramphenicol acetyltransferase gene from Vibrio anguillarum. The chloramphenicol resistant gene (cat) encoding chloramphenicol acetyltransferase (CAT) in a transferable R plasmid (pJA7324) isolated from the fish pathogen Vibrio anguillarum strain PT24 was cloned into the plasmid vector pUC19. The nucleotide sequence analysis of 1,348 base pair DNA identified an open reading frame encoding a protein of 216 amino acid residues with a calculated molecular mass of 25,471 daltons. The predicted amino acid sequences for this cat gene are 37-69% homologous with other CAT proteins of both Gram-negative and -positive bacteria. Colony hybridization performed with a PvuII-BamHI fragment including this cat gene as a probe, revealed that the same or similar chloramphenicol resistance genes existed among V. anguillarum isolates.


Focus formation by the human immunodeficiency virus (HIV) in the immobilized MT-4 cell culture and its application to the evaluation of anti-HIV agents. Immunofluorescence studies were performed on the infection of monolayer cultures of immobilized MT-4 cells with human immunodeficiency virus type 1 (HIV-1). By using the anti-viral p24 monoclonal antibody, we could observe formation of foci of p24 antigen-positive cells within 3 to 4 days when the infection was initiated with a relatively small amount of the virus. Frequency of the focus formation was in proportion to the dose of input virus (ranging from 0.001 to 0.1 PFU/cell), which allowed us to apply this phenomenon to the assay of anti-HIV agents as well as to the estimation of relative infectivity of the virus stocks. When antiviral agents were added to the infected cultures, number of foci as well as the size of each focus was reduced in a concentration-dependent manner. The dose required for reducing the number of foci by 50% was calculated to be 6 ng/ml and 8 ng/ml for tunicamycin (TM) and azidothymidine (AZT), respectively. These values are comparable to those obtained by other current assay methods. In addition, focus reduction assay is also useful in searching for such antiviral agents that would inhibit or block the early step of viral replication cycle.


An enzyme-linked immunosorbent assay using a chaotropic agent (sodium thiocyanate) for serotype specific reaction between crude dengue viral antigen and anti-dengue mouse antibody. An enzyme-linked immunosorbent assay (ELISA) has been developed to detect serotype specific reaction between crude dengue viral antigen and anti-dengue mouse hyperimmunized antibody under the stringent condition in the presence of a Chaotropic agent, sodium thiocyanate (NaSCN), in the reaction mixture of antigen and antibody. Rapidly sedimenting hemagglutinin (RHA) derived from type 2 dengue virus-infected mosquito cell culture fluid reacted to the antibody for both type 2 and type 3 dengue viruses in the ELISA. In contrast, its reactivity was reduced after the addition of NaSCN in the ELISA. Soluble complement-fixing antigen (SCF) derived from type 2 dengue virus-infected mosquito cell culture fluid reacted serotype specifically to anti-dengue type 2 antibody, and was relatively stable for the NaSCN treatment in the ELISA. Anti-type 2 RHA mouse antibody reacted to both type 1 and type 2 dengue viral antigens and its reactivity was reduced after the addition of NaSCN in the ELISA. Anti-type 2 SCF antibody reacted serotype specifically to type 2 dengue viral antigen with and without NaSCN in the ELISA.


A phage in Bartonella bacilliformis.

A phage in Bartonella bacilliformis. Bacteriophage-like particles were found in Bartonella bacilliformis culture. The particles consisted of head (icosahedral), 40 nm in diameter, and tail, 16 nm in length.


Structural similarity of the HLA-DQ region in DQ3 and DQ4 haplotypes and structural diversity of the HLA-DQ region in HLA-DR7 haplotypes. Genomic DNA obtained from a B lymphoblastoid cell line was digested with appropriate restriction endonuclease and hybridized with several probes specific for genes encoding HLA-DQ. Southern hybridization with a DQA1 3'untranslated (UT) region probe showed DQ2-type hybridization pattern in DR7DQ3 haplotype. On the contrary, DQB1 3'UT probe showed DQ3-type pattern in the same haplotype. Gene cloning and DNA sequencing analysis revealed a repetitive sequence, (TG)19, between DQA1 and DQB1 gene in the DR7DQ3 haplotype. These results suggest that a recombination event has occurred near this potential Z-DNA structure in the haplotype, DR7DQ3. The 3'UT region probes of DQA1 and DQB1 genes failed to detect restriction fragment length polymorphism (RFLP) differences between DR4DQ3 and DR4DQ4 haplotypes in this experiment, suggesting that the gene structure between DQA1 and DQB1 is conserved in these haplotypes.


Comparison of immunological effects of cholera toxin on autoimmune MRL/lpr and BXSB mice. MRL/lpr and BXSB mice were treated weekly or biweekly with cholera toxin (CT) in intravenous dose of 2 micrograms/mouse. CT treatment notably alleviated proteinuria in MRL/lpr mice, but did not influence the course of lupus nephritis in BXSB male mice. Flow cytometric analysis showed that anomalous B220+ T cells in spleen and thymus were reduced in CT-treated MRL/lpr mice while no significant change in lymphocyte populations was induced in BXSB male mice by this treatment. The suppressive effect of CT treatment on Con A response and the augmentative action on LPS response were observed in MRL/lpr mice. The latter may reflect increased B cells in relative number in the peripheral lymphoid organs. Mitogenic responses in CT-treated BXSB male mice remained unchanged in comparison with those of untreated group. Increased production of IL-6 by spleen cells was demonstrated in MRL/lpr mice treated with CT while in BXSB mice the level of IL-6 was not changed by the treatment with CT. Production of IFN gamma was suppressed by CT treatment in both strains of mice. This may be attributed to the inhibitory effect of CT on IFN gamma-producing Th1 cells as reported previously (Munoz et al, J. Exp. Med. 172: 95-103, 1990). However, CT treatment did not inhibit anti-DNA antibody production in BXSB mice, whereas the autoantibodies were markedly decreased in MRL/lpr mice treated with CT.


Further evaluation of the pregnancy-linked down-regulation of the paternal antigen-specific splenic cytotoxic T lymphocyte activity in allogeneically pregnant mice. Cytotoxic T lymphocyte (CTL) activity directed against paternal alloantigen was examined in allogeneically pregnant mice using various allogeneic combinations. The spleen cells from pregnant C57BL/6 (H-2b) mice mated with BALB/c (H-2d) male mice generated less anti-H-2d CTL after in vitro sensitization than those from unpregnant or syngeneically mated C57BL/6 mice. Different allogeneic combinations including the incompatibility at only D region of H-2 or minor histocompatibility loci were effective for downregulating the anti-paternal CTL activity in pregnancy. The downregulation of anti-paternal CTL activity induced by allogeneic pregnancy occurred at day 10 to day 18 of pregnancy, most extensively at day 14. The allogeneic pregnancy also downregulated the allogeneic CTL activities that had been amplified by injecting alloantigens before mating.


Extracellular localization of a nonfimbrial hemagglutinin of Yersinia pseudotuberculosis. A nonfimbrial hemagglutinin (HA) of Yersinia pseudotuberculosis was observed by immunoelectron microscopy using monospecific HA antiserum and protein A-gold conjugate. The HA, an amorphous but morphologically identifiable entity, was located in a region distal to or detached from the outer edge of bacteria.


[Current methods for the laboratory diagnosis of campylobacteriosis]. An analytical review of recent publications of home and foreign authors on the problem of laboratory diagnosis of campylobacteriosis is presented. The commercial nutrient media, methods of creation of the microaerophilic conditions for cultivation of campylobacter are presented. The filtration method is preferable for isolation of these agents from the studied material highly contaminated by accompanying microflora. A special attention is paid to immunodiagnosis of campylobacteriosis: agglutination reaction, coagglutination reaction, passive hemagglutination reaction, immunoenzyme and radioimmune analyses. Seroepidemiological examination of the staff at one of meat-packing factories in the Republic carried out by the method of indirect immunoenzyme analysis has revealed high levels of anticampylobacteriosis antibodies in 17.9% of examinees. The promising trends in perfection of the methods for laboratory diagnosis of campylobacteriosis are outlined.


[The effect of physicochemical factors on the activity of DNA-dependent DNA polymerases in Acholeplasma laidlawii PG-8]. The biological and physico-chemical properties of DNA-dependent DNA-polymerases of Acholeplasma laidlawii PG-8 have been studied. The optimal parameters of maximal enzymatic activity are determined. It is stated that N-ethylmaleimide in concentration of 1 mM activated DNA-polymerase I by 52%, whereas DNA-polymerase II with reagent concentration of 0.5 mM demonstrated the peak of activity exceeding the control only by 10%. Spermidine in concentration of 1.5 mM for the first form of DNA-polymerase and 0.15 mM-for the second one increased the ability of both forms of polymerases to synthesize DNA by 10%. Aphidicolin added to the reaction medium up to concentration of 10 mg/ml decreased activity of forms I and II of enzymes by 83 and 68%, respectively. The presence of 0.6 mM of EDTA in the medium also negatively affected the activity of polymerases inhibiting it by 83% in form I and by 77%-in form II.


[The RNAse activity in mycoplasmas of different taxonomic positions]. Dynamics of the RNAase accumulation in cells and culture fluid of strains of Acholeplasma and Mycoplasma genera has been studied. In certain strains of two genera of mollicutes some differences in dynamics of RNAase formation by them are determined.


[The effect of physicochemical factors on the growth of Pseudomonas putida BS-2 on a medium with diethylene glycol]. The physicochemical factors of medium have been studied for their effect on the physiological indices of growth of Pseudomonas putida BS-2 culture utilizing diethylenglycol as the only source of carbon. Action of the supraoptimal temperature on the growth process of P. putida BS-2 is accompanied by a decrease (more than twice) in economic coefficient of substrate and specific growth rate as compared with their maximal values. Dependences of specific growth rate of P. putida BS-2 in the medium with diethylenglycol on the presence of NaCl in it within the range of its concentrations from 0 to 4% and methanol in the concentration range of 0-20 g/l follow the noncompetitive inhibition equation. When NaCl concentration in the medium is more than 4%, complete separation of constructive and energy metabolism processes is observed.


[The immunological shifts in the blood sera of persons inoculated with influenza vaccines]. The dynamics of antihemagglutinin content in blood serum of students inoculated with different types of influenza virus vaccines has been traced out in the process of long-term observation. It is shown that this dynamics depends on the type of used vaccines, method of their introduction and epidemiological situation. It is stated that in the process of annual vaccination the highest titres of antihemagglutinins are observed in twice inoculated patients; after the third vaccination these titres decrease and after four-fold vaccination--they increase again.


[The intestinal microflora of persons subjected to a radiation lesion]. The content of large intestine has been studied in persons exposed to radiation injury in consequence of the accident at the Chernobyl Atomic Power Plant. It is stated that bifidobacteria (10(7)-10(10) cells in 1 g of feces) prevailed (as in healthy people), Bifidobacterium indicum being a dominating species. Amount of lactic-acid bacteria in 1 g of defecations of examined patients was within the range of 10(6)-10(9) cells and in certain persons it reached 10(10) cells (primarily fecal Enterococci). A considerable amount of patients with acute radiation sickness of the 3d degree had in their intestine 10(9)/g of lactic-acid bacteria, Lactobacillus casei and L. plantarum prevailing there. The frequency of yeast isolation from defecations of patients constituted 83%, while the number of cells in 1 g of feces--from 10 to 10(4). Yeast of the Candida genus, mainly Candida parapsilosis, prevailed. The species composition of isolated microorganisms has no substantial differences from microcenosis of healthy people. The content of intestine of persons suffered from radiation is characterized only by greater amount of lactic-acid bacteria and enterococci as compared with healthy adults.


[Bacteria of the genus Aeromonas and their role in aquaculture]. Bacteria of genus Aeromonas are constant components of microbiota of fresh reservoirs where they, together with other microorganisms, play the part of natural biofilter and promote water self-purification. They are necessarily present in normal microflora of hydrobionts inhabiting fresh reservoirs. The greatest attention is paid by the researchers to Aeromonas and biotrophs in connection with epizootics in aquaculture which have become more frequent, in particular, under fish breeding. That is why the review is, to more extent, concerned in the works of this trend made by the foreign and home researchers for the last decade.


Community prevalence survey of children's blood lead levels and environmental lead contamination in inner Sydney. To determine the distribution of blood lead levels in preschool children in inner Sydney and identify possible sources of environmental lead. Cross sectional community based prevalence survey of children and the houses in which they live, and a survey of volunteer children. Mort Bay and Summer Hill, residential localities in inner Sydney. Ninety-five children aged 9-48 months able to be identified in a defined geographic area and 63 children aged 9-48 months volunteered by their parents. Concentrations of lead in venous blood of all children and in samples from the home environment of Mort Bay children. Four of the children (2.5%) had blood lead levels > or = 1.21 mumol/L (25 micrograms/dL, the current Australian threshold of concern), 27 (17.1%) had levels > or = 0.72 mumol/L (15 micrograms/dL, the new US threshold for individual intervention) and 80 (50.6%) had levels > or = 0.48 mumol/L (10 micrograms/dL, the new US threshold for community intervention). Blood lead concentrations were significantly correlated with concentrations of lead in "sink" soil (r = 0.555, P = 0.026), play area soil (r = 0.492, P = 0.016) and dust from vacuum cleaners (r = 0.428, P = 0.05), and with age of child (r = -0.182, P = 0.023). The presence of the child during house renovation was a strong predictor of having a blood lead level above 0.72 mumol/L (15 micrograms/dL) (odds ratio, 4.6; 95% confidence interval, 1.8-11.7, P = 0.001). Lead in soil and in household dust in older areas of Sydney is likely to represent a significant health hazard to young children. Many thousands of children may be affected in Sydney and other Australian cities. There is an urgent need for expanded prevalence surveys, public education and the development of strategies for the abatement of lead in urban environments.


The use of portable blood glucose monitors by trained lay operators. To evaluate the use of Reflolux II blood glucose reflectance meters by trained lay operators in a community survey. A random household survey involving assessment of participants' blood glucose levels by Reflolux II. For a random subsample of participants an additional sample of blood was placed onto filter paper strips and stored for later examination and assay. The survey was conducted in the Newcastle and Lake Macquarie areas of New South Wales. Of the 1275 participants 15 years and over, 1229 consented to have their blood glucose assessed and 163 of these also had blood stored on filter paper strips. Data were collected by 25 interviewers. Application of blood spots to filter paper assessed the interviewers' capacity to collect samples of minimal acceptability for Reflolux II operation. Samples were considered adequate if they covered at least a 6 mm soaked disk and there was no evidence that they had been smeared or applied from both sides of the paper. The Reflolux II readings obtained by the interviewers were compared with the blood glucose values from assay of the filter paper samples. Only 63% of the 163 samples collected on filter paper were of acceptable quality for adequate Reflolux II operation. The overall correlation between Reflolux II blood glucose values and those determined from the blood collected on filter paper was good (r = 0.893). However, three of the eight interviewers who had five or more readings for comparison achieved correlation coefficients of < r = 0.20. Of the survey participants identified as having elevated blood glucose levels (> 8 mmol/L) by the filter paper assay, 58% (7 of 12) were not detected by the lay operated reflectance meters. Twenty-two per cent of the individuals assessed by the reflectance meter, compared with 0.5% of a population sample surveyed by the National Heart Foundation, had blood glucose values of less than 3.5 mmol/L. It would appear that there may be a considerable error rate in blood glucose values obtained by lay operators using reflectance meters, resulting in underestimation of blood glucose levels. The findings have implications for community awareness programs for diabetes and highlight the need for careful training and monitoring of lay operators in their obtaining of finger-prick blood samples and their use of reflectance meters.


Regional variation in coronary mortality within Tasmania. To measure regional rates of mortality from ischaemic heart disease (IHD) within Tasmania and to analyse factors associated with regional differences. Descriptive epidemiological study. Community-based study. Male residents of Tasmania aged 30-69 years dying from IHD between 1986 and 1989. Coronary death as coded by the Australian Bureau of Statistics and place of death validated by hospital and community data. This study identifies substantial differences in coronary death rates among the three Health Regions of Tasmania. These differences are real and are not caused by variations in diagnostic or coding practices between regions. The two northern Health Regions, which represent approximately 52% of the total population, account for 98% of the excess mortality from IHD in Tasmania compared with the national rate. More detailed analysis of these differences suggests that variation in the number of deaths occurring in hospital contributes significantly to the regional differences in death rate from IHD. Rates of coronary mortality in Tasmania have been significantly higher than in all other Australian States for much of the past decade because of a higher death rate within the population of the northern half of Tasmania. Differences in mortality rates between regions in Tasmania provide a focus for further study into the causes of the unacceptably high rates of death from IHD in Tasmania and underline the need for the funding of a coronary register in Tasmania.


The importance of hypertension in the aetiology of infarctive and haemorrhagic stroke. The Lower Hunter Stroke Study. To determine the importance of hypertension in the aetiology of infarctive and haemorrhagic stroke in persons aged 35-69 years. A population-based case-control study. Lower Hunter Region community. One hundred and ninety patients with a first stroke were identified from a register, including all hospital admissions and death certificates in the Region, and compared with 496 control subjects obtained from a random population sample of the same community. First event of stroke (fatal or non-fatal). Twenty-seven per cent of those with a haemorrhagic stroke, compared with 2% of those with infarctive stroke, died before hospital admission; the in-hospital mortality was 15% and 9%, respectively. Twenty-one per cent of control subjects, compared with 51% of those with stroke, were currently receiving treatment for hypertension. By logistic regression analysis the odds ratio for receiving current treatment for hypertension in those with haemorrhagic stroke was 5.5 (95% confidence interval [CI], 2.36-12.8), compared with 2.53 (95% CI, 1.48-4.34) in those with infarctive stroke. Other differences between haemorrhagic and infarctive stroke included no excess risk in men for haemorrhagic stroke but an odds ratio of 3.51 (95% CI, 1.83-6.74) for infarctive stroke; and a steep risk gradient for obesity in haemorrhagic but not in infarctive stroke. Cigarette smoking carried a non-significant odds ratio of around 1.5, with no difference between stroke type. Among those who had ever been told that they had hypertension, 75% and 71% of patients with infarctive stroke and haemorrhagic stroke, respectively, and 61% of control subjects, were currently receiving treatment for hypertension. In those stroke patients who were currently being treated for hypertension, 63% had a pre-admission diastolic blood pressure of 90 mmHg or more. The mean diastolic blood pressure levels on admission were 10 mmHg higher than the latest recorded pre-hospital measurements and fell to 10 mmHg lower than the levels recorded before hospital admission by the time of discharge. Hypertension is important in the aetiology of both infarctive and haemorrhagic strokes, although it may be more important in haemorrhagic stroke, and there appear to be other aetiological differences between stroke types. Most of the patients with a history of hypertension were currently receiving treatment for hypertension, although blood pressure control before admission was not optimal.


Successful management of primary hypercholesterolaemia with simvastatin and low-dose colestipol. To examine whether a small dose of bile acid sequestrant used in combination with a hydroxymethylglutaryl coenzyme A reductase inhibitor is more effective in reducing serum and low-density lipoprotein (LDL) cholesterol levels than inhibitor used alone. A randomised, double-blind study. Subjects receiving tertiary care at a hospital lipid clinic. Subjects with severe primary hypercholesterolaemia (types IIa and IIb), already stabilised on a cholesterol-lowering diet, with serum cholesterol levels of 7.0 mmol/L or more and triglyceride levels of 6.0 mmol/L or less. Sixty-four subjects were randomly assigned to the treatment groups; three withdrew before any outcome observations; 61 completed the trial and their results were analysed. Subjects were randomly assigned to receive either colestipol placebo or colestipol 5 g or 10 g each morning in fixed dosage for 18 weeks. They simultaneously received incremental doses of simvastatin: placebo for six weeks, then 20 mg/night for six weeks, then 40 mg/night for a final six weeks. Lipids, lipoproteins, and haematological and biochemical safety parameters were measured at the end of each treatment period. Adverse events were monitored. Respective maximum reductions (95% confidence intervals) in serum cholesterol, LDL cholesterol and apolipoprotein B (apo-B) values in subjects taking combination therapy were 41% (38%-45%), 50% (46%-53%) and 43% (39%-46%), compared with lesser reductions of 32% (26%-37%), 38% (31%-45%) and 37% (32%-41%) in those taking simvastatin monotherapy. The percentage changes in LDL cholesterol with combination therapy were independent of baseline cholesterol level or lipid phenotype. Combination therapy reduced serum triglyceride levels by up to 24% (15%-32%) and increased high-density lipoprotein (HDL) cholesterol levels by up to 9% (3%-15%). Three subjects withdrew within a few weeks because of severe gastrointestinal side effects related to colestipol; 19 experienced milder gastrointestinal side effects, 15 were taking combination therapy. A combination of low-dose colestipol and simvastatin was found to be more effective in reducing serum and LDL cholesterol than simvastatin used alone. Such combination therapy offers the possibility of improved cholesterol lowering without the need for full dosage of either drug.


Adverse reaction to a Chinese herbal remedy. To report a multisystem illness in a patient after ingestion of Chinese herbal medicines, together with an analysis of the herbal medicine and to alert physicians to the growing clinical importance of adverse reactions to herbal remedies. A 61-year-old Chinese Vietnamese man with a history of atrial fibrillation and left cerebral hemisphere infarction three months previously presented after a three-day illness with acute hepatorenal failure, multiple pulmonary emboli, peripheral arterial thromboembolism and laboratory features of consumptive coagulopathy. For three days before presentation, he drank a daily infusion of herbal medicine. Intensive supportive medical care including haemodiafiltration resulted in recovery of renal function, liver function and return of coagulation parameters to normal over a one-month period. Toxicological analysis of an infusion prepared from a sample of the patient's herbal medicine revealed the potentially toxic compounds benzaldehyde, cinnamoyl alcohol and ephedrine. The ingestion of herbal medicine caused or contributed to the patient's illness.


Behavioural changes due to pontine and extrapontine myelinolysis. To report a case of central and extrapontine myelinolysis with unusual behavioural manifestations that developed after slow (not more than 0.5 mmol sodium per litre per hour) correction of hyponatraemia. A 51-year-old Caucasian woman with hyponatraemia caused by psychogenic polydipsia experienced a delayed onset of behavioural changes with some ataxia, but no pyramidal signs, after correction of the hyponatraemia. The hyponatraemia was corrected with a combination of hypertonic saline and fluid restriction at the currently recommended rate of not more than 0.5 mmol of sodium per litre per hour. Central and extrapontine myelinolysis may develop after slow correction of hyponatraemia. Behavioural manifestations may be the most prominent clinical feature.


Will computers replace or complement the diabetes educator? Computer programs are playing an increasingly important role in the management of diabetes and fall into various categories. Instructional programs are used to teach patients about diabetes. They cover the principles of blood glucose monitoring, diet, exercise, foot care and so on. Statistical and graphical analysis programs are used in diabetes clinics to help the physician, diabetes educator and patient detect patterns and trends in the patient's home blood glucose readings. Hand-held insulin dosage computers have been used by patients to advise them on insulin dosage adjustment on a day-to-day basis. The diabetes simulator we have recently developed allows patients to practise, and gain experience with insulin dosage adjustment; the patient decides on alterations to insulin dose, diet or the amount of exercise and the computer program demonstrates the resulting effect on blood glucose levels. Overall, computers complement and enhance, rather than replace the functions of the diabetes educator.


Treatment of primary hypercholesterolaemia with pravastatin: efficacy and safety over three years. To assess the efficacy, safety and tolerability of pravastatin over three years of treatment. An open, multicentre randomised study. Subjects receiving tertiary care at three hospital lipid clinics. Subjects with primary hypercholesterolaemia (type IIa) or combined hyperlipidaemia (type IIb), already stabilised on a cholesterol-lowering diet, with low density lipoprotein (LDL) cholesterol levels of greater than 4.7 mmol/L and triglyceride levels of less than 4.5 mmol/L. Sixty-one subjects were randomly assigned to the treatment groups: 60 completed 12 weeks and 46 completed 30-36 months of treatment. Subjects were randomly assigned to receive either pravastatin 20 mg/day, pravastatin 40 mg/day or cholestyramine 16 g/day for a period of 12 weeks. Subsequently, dose titration of pravastatin up to 40 mg/day was permitted, if required, and all groups received supplementary therapy with other lipid-lowering drugs. Lipids, lipoproteins, haematological and biochemical safety parameters were measured at regular intervals. Adverse events were monitored. There were significant reductions in total and LDL cholesterol levels with all treatments over 12 weeks (P < 0.001). The mean reductions (+/- SD) in LDL cholesterol were 26% +/- 14% in the group taking pravastatin 20 mg/day (n = 21), 30% +/- 8% in the group taking pravastatin 40 mg/day (n = 21) and 34% +/- 13% in the group taking resin (n = 18). The percentage changes in LDL cholesterol were independent of age, baseline cholesterol level or lipid phenotype. High density lipoprotein (HDL) cholesterol levels were significantly increased, by 8%-18% with all treatments (P < 0.001). Triglyceride levels were reduced by high-dose pravastatin only (7% +/- 29%), but were found to increase with resin (45% +/- 63%). During long-term treatment over 36 months, still greater reductions in total and LDL cholesterol were found in patients taking pravastatin (n = 35), but not in those taking resin (n = 11). There was an apparent decrease in effect beyond 18 months in both groups, possibly related to reduced compliance with diet or cholestyramine intake. Eight subjects allocated to pravastatin and seven allocated to resin withdrew (one and two subjects respectively because of drug-induced adverse events). Adverse events during 12 weeks' monotherapy with pravastatin included central nervous system (CNS) symptoms (12%), gastrointestinal (GIT) symptoms (7%) and an acute hepatitic reaction (one subject). Of those in the resin therapy group, 22% developed GIT symptoms. Myalgia occurred in three subjects using a combination of pravastatin and clofibrate, but this resolved fully upon clofibrate withdrawal. Pravastatin was found to be a relatively effective, safe and well tolerated lipid-lowering drug. Still greater LDL reduction was achieved with pravastatin combination therapy and this was essentially maintained over three years.


Pulmonary angiography in a non-teaching hospital over a 12-year period. To report the safety of pulmonary angiography in a non-teaching hospital and discuss the place of this investigation in the diagnosis of pulmonary embolism. A retrospective review of all patients undergoing pulmonary angiography over a 12-year period, November 1979 to October 1991, at Dandenong Hospital, Melbourne. A protocol was established whereby each procedure was performed with the assistance of staff and equipment from the intensive care unit to provide haemodynamic monitoring and measurement of pulmonary artery pressures. During the review period 114 patients underwent pulmonary angiography, most (108) for suspected pulmonary embolism. One hundred and fifteen pulmonary angiograms were performed, and no deaths related to the procedure occurred. Complications included perforation of the right atrium in three patients, with no sequelae. Cardiac arrhythmias were common but self-limiting. In our series, pulmonary angiography was a safe procedure if undertaken with the appropriate monitoring measures. A protocol which includes the participation of the intensive care unit has proved helpful. In addition, it was found that clinical symptoms and signs were unreliable and in 24 patients who also had ventilation-perfusion (V/Q) scans, only seven scans were accurate. A review of the literature indicates that V/Q scans lack specificity and that pulmonary angiography may be performed safely. The latter investigation should be more frequently performed.


Exposure to blood borne infections in health care workers. To determine the incidence and nature of occupational exposures to blood and body fluids in health care workers. 332 reports of occupational exposure were analysed and are presented. A major teaching hospital. All staff at Royal Perth Hospital who reported an occupational exposure to blood or body fluids to the Department of Clinical Immunology between 1 January 1990 and 31 August 1991. The rate of reported occupational exposure according to staff category, nature of exposure, HIV status of source patient, activity at the time of exposure and compliance with infection control measures. 332 reports from 323 health care workers were received, giving an overall incidence of 6.1 per 100 full time equivalent (FTE) years. Nursing staff (9.4/100 FTE years) and medical staff (9.0/100 FTE years) reported exposure more frequently than housekeeping staff (2.5/100 FTE years) or paramedical staff (2.3/100 FTE years) (P < 0.001). The rate of exposure to HIV antibody positive patients was only 0.24/100 FTE years. Needlestick or other blood contaminated sharps injuries accounted for 83.4% (277/332) of reports and failure to observe universal precautions for 34.0% of reports. Insertion and operation of parenteral lines (24%) and performing operations (15.4%) were the activities most often associated with occupational exposure. No occupationally acquired infections were observed. Despite the immediate availability of zidovudine, acceptance by health care workers with high risk occupational exposure was low (18.8%). Occupational exposure to blood and body fluids is common among health care workers but most exposures confer a low risk of blood borne infection. The introduction of an occupational exposure assessment program has many benefits, including optimal management of injuries and acquisition of data on infection control measures, and may protect health care institutions from false claims for compensation.


Mycobacterium marinum infection: epidemiology and presentation in Queensland 1971-1990. Evaluation of the clinical and epidemiological features of Mycobacterium marinum infection in Queensland. Laboratory identification and in-vitro susceptibility tests of 29 isolates from the Queensland Health Department Tuberculosis Reference Laboratory were retrospectively gathered and followed up by contacting referring practitioners and obtaining clinical details of patients involved. 29 patients from whom M. marinum was isolated, with a male:female ratio of 3.1:1, and a mean age of 47.4 years. Of 26 patients for whom adequate information was available, 12 had evidence of involvement of deep tissues (including two cases of arthritis) and five suffered sporotrichoid spread of infection. The delay between onset of symptoms and consultation with a medical practitioner was five months (range, two weeks to two years), with a further mean delay to definitive diagnosis of 4.4 weeks. Cure was apparent in 22 of 23 cases. Chemotherapy alone was adequate in 11 cases, as was surgical intervention in three, while a combination approach was successful in eight cases. Trimethoprim/sulfamethoxazole was successful in seven of nine cases and combination rifampicin and ethambutol in six of seven. Tetracyclines were employed as single-agent therapy in nine patients and were effective in seven. Synovitis was a common presenting feature of M. marinum infection in Queensland patients. Occupational and recreational exposure to salt or fresh water was common, and although this history was available to practitioners a mean delay to definitive diagnosis of 4.4 weeks still occurred. The data suggest that chemotherapy alone is often adequate, even with deep tissue involvement. Combinations of conventional antimycobacterial drugs may be the therapy of choice, especially for serious infections, although success was recorded with trimethoprim/sulfamethoxazole alone.


The economic impact of chronic fatigue syndrome. To estimate the economic impact of chronic fatigue syndrome (CFS) on the individual, the government, and the community. The financial burden produced by CFS was studied by calculating the direct and indirect costs arising from the disorder. Data regarding use of health resources, income and employment were obtained by questionnaire from patients with CFS. In addition, aggregate Medicare data on the incidence and fees charged for each Schedule item for these patients was obtained. The Richmond Valley, New South Wales. Forty-two patients with CFS identified in our population-based prevalence study. The conservative estimate of the per annum costs of CFS in the Richmond Valley, with a prevalence of 37.1 cases per 100,000, was $396,000. If extrapolated to the Australian population, we estimate CFS would generate an annual cost of at least $59 million. This disorder constitutes a large but neglected area of health resource utilisation and economic burden.


Modern methods of searching the medical literature. To review the methods of electronic retrieval of biomedical journal articles; to demonstrate CD-ROM and online searching techniques; and to compare major biomedical databases. A CD-ROM MEDLINE search was made by a medical practitioner, with the assistance of an experienced librarian. An online MEDLINE/Excerpta Medica search was made by the same medical practitioner after completion of a two-day course in search techniques. The search example covered the period 1983 to 1992, and the topic was "whether steroid use for asthma/airways disease leads to osteoporosis". Search duration, article yield and search cost were assessed. The CD-ROM MEDLINE literature search took 60 minutes to find 31 articles (43% of the possible articles). This compared with an online search to the Bibliographic Retrieval Services database in Chicago, which produced the same articles when MEDLINE was searched (but took only 16 minutes) and which also found 47 articles (with an overlap of only seven) when Excerpta Medica was searched. We estimated that the combined MEDLINE and Excerpta Medica search detected 93% of all known relevant articles, based upon an assumed gold standard. Online searching is a fast and powerful alternative to CD-ROM searching. Formal training is necessary for quick and cheap reference finding. Other benefits such as direct transfer from online searching to your own computer bibliography database make online searching of biomedical databases essential to consider.


Breaking bad news.

Breaking bad news. To investigate the importance of a sympathetic approach to breaking bad news in the doctor-patient relationship. Worldwide surveys of views on truth disclosure by health care professionals, the public, and patients with cancer and other diseases. Surveys using a Medline Computer Search were identified. Twenty-two questionnaire studies. The data obtained demonstrate trends of an increasing wish since 1950 on the part of patients, the public and doctors for more openness. The skills necessary for breaking bad news well can be acquired through organised undergraduate and postgraduate education which emphasises a good working doctor-patient relationship.


Transplantation of the endocrine pancreas in 1992. In 1992, transplantation of the endocrine pancreas is placed at the interface between laboratory research and clinical medicine. Unlike other forms of transplantation, it is not life saving, and yet its potential to treat all patients with insulin dependent diabetes is limited only by the availability of donated organs. Arguably, the long term glucose homoeostasis provided by vascularised pancreas transplantation is the standard by which other forms of endocrine pancreas transplantation (islet cell, fetal islet, and xenogeneic transplantation) must be judged. For the small number of type I diabetic Australians who develop end-stage renal failure and would otherwise require a renal transplant, the combined transplant procedure is a technically viable treatment option and, with some justification, the treatment of choice.


Why some people do not drown. Hypothermia versus the diving response. To examine our present state of knowledge regarding the remarkable survival of some victims from prolonged submersion for an hour or more. Debate continues on the relative importance of the two explanatory theories--diving response and hypothermia. A wide range of physiological, pathophysiological and clinical papers relating to the diving response, hypothermia and near-drowning, with emphasis on the period 1981-1991. The normothermic human brain suffers irreversible damage if subjected to acute asphyxia for longer than 10 minutes. Significant resistance of brain tissue to hypoxia occurs only after its temperature has fallen from 37 degrees C to 30 degrees C or less. Body surface cooling depresses core temperature by only one-third of this drop in 10 minutes. Hence an additional factor, other than hypothermia, is required to explain survival from near-drowning. The idea that ingestion and aspiration of large amounts of cold water produce such a temperature drop lacks quantitative evidence. The diving response seen in marine mammals also occurs in humans but to a lesser extent; however, about 15% of volunteers tested exhibit a profound response. This response which starts immediately upon submersion prevents aspiration of water, redistributes oxygen stores to heart and brain, slows cardiac oxygen use and initiates a hypometabolic state. The possible influence of alcohol on these processes is considered. Survival from prolonged near-drowning appears to depend upon a specific temporal interplay between the diving response and hypothermia, resulting in a protective state of hypometabolism.


Long-term neonatal rat aortic smooth muscle cell cultures: a model for the tunica media of a blood vessel. Cultured neonatal rat aortic smooth muscle cells have been used to study the synthesis and accumulation of extracellular matrix components in many laboratories. These cells are capable of accumulating large amounts of insoluble elastin in the extracellular matrix and can be maintained in culture for long periods of time without subcultivation. This study examined the elastin and collagen contents of such cells in culture for 5, 21 and 43 weeks. The percent elastin and collagen observed in the 43-week cultures were strikingly similar to that seen in the intact neonatal rat aorta. It should be noted that the percent collagen varied significantly between 5 weeks and 43 weeks, whereas that for elastin remained relatively constant throughout the same time course. Histological examination demonstrated that the elastin fibers in the extracellular matrix of the cultures were arranged in a pattern similar to the elastic lamellae of the aortic tunica media. Data presented here suggest that these cells in culture mimic the donor tissue from which they were derived with respect to elastin and collagen content as well as elastic fiber arrangement, and possibly represent an organotypic culture of the medial layer of a blood vessel.


Production, purification and characterization of canine prostromelysin. One of the best studied animal models of osteoarthritis is a dog model in which the anterior cruciate ligament of the hind limb stifle joint is transected (Pond-Nuki model). To determine whether stromelysin might play a role in this model, it was necessary to purify the enzyme for production of suitable probes. In the present study, dog synovial fibroblasts were stimulated to express a metalloproteinase that was demonstrated to be canine prostromelysin by Northern blot, protein purification and amino-terminal sequence analyses. Unlike rabbit synoviocytes, passaged dog synoviocytes did not express stromelysin mRNA in response to recombinant human IL-1, but expressed stromelysin mRNA only upon treatment with dog monocyte-conditioned medium (dMCM). The aminophenylmercuric acetate (APMA)-activatable metalloproteinase present in the culture supernatants of stimulated dog synoviocytes was purified using a combination of ion-exchange and dye matrix affinity chromatography. The purified canine metalloproteinase co-migrated on reducing SDS-PAGE with recombinant human prostromelysin-1 as a doublet with apparent molecular masses of 54 and 56 kDa. Similar to APMA-activated human prostromelysin-1, the APMA-activated canine metalloproteinase was inhibited by 1,10-phenanthroline or recombinant human tissue inhibitor of metalloproteinase (TIMP). The amino-terminal sequences of the canine pro- and APMA-activated enzymes were compared with those of human, rabbit and rat stromelysin. The striking homologies among the sequences demonstrated that the purified canine metalloproteinase was indeed canine prostromelysin. A rabbit anti-canine prostromelysin polyclonal antiserum was generated and used to localize the enzyme within cultured dog synoviocytes and articular cartilage stimulated with dMCM. The reagents developed in this study should be useful for examining the expression and distribution of prostromelysin in canine models of osteoarthritis.


Dentin phosphophoryn binding to collagen fibrils. The interaction of rat incisor phosphophoryn with native turkey tendon collagen fibers has been examined by electron microscopy. The binding of phosphophoryn to the tendon fibril surfaces is quite selective. The phosphophoryn is seen as positively or negatively stained globular particles predominantly at the "e" band in the collagen gap region in transmission electron micrographs of the phosphophoryn-reacted fibrils. The selectivity of binding to the fibrils was obtained in the presence of calcium ions, which bind avidly to phosphophoryn. The specific association of phosphophoryn at the "e" band suggests a possible regulation of mineral deposition within the gap regions of the collagen fibrils.