Determination of gold-based antiarthritis drugs and their metabolites in urine by reversed-phase ion-pair chromatography with ICP-MS detection. A sensitive method for the determination of gold-based drugs auranofin, myochrysine, and their metabolites has been developed. These gold-containing compounds were separated by reversed-phase ion-pair chromatography with tetrabutylammonium chloride as the ion-pairing agent. Gold-specific on-line detection utilized inductively coupled plasma mass spectrometry (ICP-MS). The separation conditions of pH, methanol content, concentration of the ion-pairing agent and ionic strength have been investigated. The detection limit for auranofin, the last peak in the chromatogram, was 0.3 ng. These methods were applied to the analysis of gold-containing species in urine from arthritis patients on auranofin, myochrysine or solganol therapy. The recovery of the total gold-containing species from urine was greater than 90%. Dicyanogold(I) anion, [Au(CN)2]-, was detected in the urine of several patients.


Direct injection analysis of carbamazepine and its active 10,11-epoxide metabolite in plasma by use of a semipermeable surface (SPS) silica column in LC. A semipermeable surface (SPS) silica column was applied for the simultaneous determination of carbamazepine (CBZ) and its active 10,11-epoxide metabolite (EPO) in plasma following direct injection in LC. The SPS packing material consists of an ODS ligand as the hydrophobic inner phase and a polyoxyethylene network as the hydrophilic outer phase. When a 5-microliters portion of intact plasma was injected onto the column using a mobile phase of phosphate buffer (pH 7.1, ionic strength 0.1)-acetonitrile (4:1, v/v), the plasma proteins were size-excluded, whereas the drug and its metabolite were retained and separated both from each other and from other commonly co-administered drugs such as phenobarbital (PB) and phenytoin (DPH). The calibration graphs (peak area vs concentration) of CBZ, EPO and PB were linear over the therapeutic range of plasma concentration (r greater than 0.998) with good relative standard deviations (RSD less than 3.98%, n = 5). The recoveries from plasma were almost complete (greater than 96.6%). The analysis time was 17 min. The method as developed was applied in studies on the time course of plasma concentrations of unchanged CBZ and EPO after i.v. administration of CBZ to the rat.


Analysis of [14C]-saterinone and its metabolites in dog plasma, urine and bile by high-performance liquid chromatography with radioactivity and fluorescence detection and by mass spectrometry. An HPLC method was developed for the direct on-line radioactivity determination of [14C]-saterinone and its metabolites in biological canine fluids after intravenous and intraduodenal administration. After direct injection of 200 microliters of sample, the metabolites were separated on a semi-preparative reverse-phase column. The metabolites were identified by HPLC reference standards, enzymatic hydrolysis and mass spectrometry. Besides a small amount of unchanged saterinone, six metabolites could be detected, both in bile and urine. The main fraction (about 80-90% of the sum of detected metabolites) contained the phase II metabolites of saterinone, the sulphate and glucuronide. Ring hydroxylated saterinone and three metabolites that were not identified made up about 1-4% each. In plasma, only the major compounds could be detected because of the lower absolute concentrations. The metabolic pathway of saterinone in dogs was elucidated and compared with other species. Results from previous studies concerning a first-pass metabolism could be confirmed.


Rapid determination of conditional partition constants in an FIA system. This paper presents a simple and rapid method for the screening of substances for their conditional partition constants in the chloroform-water system. Samples are injected in either aqueous or organic solution into the corresponding phase. After equilibration in segmented flow the amount of sample remaining is measured photometrically. Performing the measurements at a series of phase volume ratios enables calculation of conditional partition constants knowing neither the amount injected nor the absorptivity of the substance under study. The time needed for a determination is about 15 min per substance, in many cases even shorter and the sample consumption is, in most cases, less than 1 mg. These characteristics make the method suitable for screening purposes. A series of compounds has been examined with this method, and acceptable results for the conditional partition constants were obtained in the range 0.2 less than D' less than 100 (-0.7 less than log D' less than 2.0).


Determination of thevetin B in serum by fluorescence polarization immunoassay. Thevetin B, a cardiac glycoside of Thevetia neriifolia Juss. seeds, was determined in serum by fluorescence polarization immunoassay. Anti-digitoxin antibody was used, thevetin B genin being structurally identical to digitoxigenin. Cross-reactivity of 94% was found by this method, for concentrations from 5 to 80 ng ml-1.


Colorimetric and titrimetric assay of isoniazid. Two methods are proposed for the determination of isoniazid in pure form or in tablets. In the first method chlorpromazine hydrochloride, when treated with 2-iodoxybenzoic acid as an oxidant in 50% w/v o-phosphoric acid solution, is oxidized to chlorpromazine free radical which absorbs at 530 nm. The red free radical is readily reduced quantitatively by isoniazid to the colourless chloropromazine. The addition of isoniazid to a red solution of chlorpromazine free radical results in a decrease in absorbance in direct proportion to the quantity of isoniazid. This forms the basis for the quantitative determination of micro-quantities of isoniazid (3-18 micrograms ml-1). The second method involves the titrimetric determination of isoniazid using N-bromophthalimide as a titrant. The end-point is determined either directly using methyl red or amaranth as indicator, or by a back titration method in which a known excess of N-bromophthalimide solution is added to isoniazid solution and then the residual unreacted reagent is determined iodometrically. The results by the proposed procedures were in good agreement with those obtained by the official methods.


Determination of 9-hydroxyellipticine by redox colorimetry. The chemical reactions involved in the decomposition of 9-hydroxyellipticine (9-OH-E), an anticancer agent, in polar solvents is explained. The reactions, which involve the formation of 9-oxo-ellipticine and the addition of a nucleophilic acid on the C10 site of the heterocyclic system, have been used to measure 9-OH-E quantitatively by colorimetry in solution and by reflection on paper surfaces. A method for the stabilization of 9-OH-E in polar solvents is proposed.


The identification of related substances in 9 alpha-fluoroprednisolone-21 acetate by means of high-performance liquid chromatography with diode array detector and mass spectrometry. A method for the analysis and identification of the principal related substances in 9 alpha-fluoroprednisolone acetate is described. This compound has been chosen as a model for the investigation of related substances which can be originated in the general procedure for introducing a fluorine substituent at position 9 of a corticosteroid molecule. HPLC procedures, both in reversed and in normal phase were used; a rapid scanning UV detector which allows direct spectrophotometric data to be obtained on chromatographic peaks, proved to be a tool of great importance. Thus, after reversed-phase chromatographic separation and observation of the UV spectra and their respective second derivatives, it was possible to characterize some of the principal effective and potential related substances such as 9 alpha-fluorohydrocortisone acetate, 9 alpha-bromoprednisolone acetate, 9 beta, 11 beta-epoxyprednisolone acetate and 9(11)-dehydroprednisolone acetate, emerging as chromatographic peaks. Identification of 9 alpha-bromoprednisolone acetate and of 9 alpha-fluorohydrocortisone acetate which proved to be the most significant impurities, was confirmed by means of an exhaustive study of the mass spectra of these substances conveniently isolated by normal-phase HPLC. The chromatographic, spectrophotometric and mass-spectrometric characteristics of the studied compounds are reported.


Liquid chromatographic resolution of the isomers of tipredane and phenylthioproline using urea-solubilized beta-cyclodextrin in the mobile phase. Two LC assays were developed using urea-solubilized beta-cyclodextrin (beta-CD) as a mobile phase additive in combination with reversed-phase columns. A methylsilane column gave optimal resolution of the steroid tipredane from its epimer. An investigation of the effects of beta-CD concentration, column temperature, column type, and addition of ethanol on the chromatographic separation is detailed. The enantiomer and diastereoisomers of L-cis-phenylthioproline were best resolved by urea-solubilized beta-cyclodextrin and a trimethylsilane column. The elution order of L-cis-phenylthioproline relative to its stereoisomers was reversed after adding ethanol to the beta-CD containing mobile phase or by changing from a beta-CD to an acetylated beta-CD column. The resolution factors for these separations obtained using the beta-CD mobile phase were larger than those obtained using beta-CD columns. Mobile phases containing up to 0.15 M beta-CD and 8 M urea were investigated. The separation of these isomers are dramatically affected by column polarity.


A competitive enzyme immunoassay for the direct determination of mometasone furoate (SCH 32088) in human plasma. Mometasone furoate (SCH 32088) is a synthetic corticosteroid which has a topical anti-inflammatory activity with a minimal potential for suppressing hypothalamic-pituitary-adrenocortical (HPA) axis. A sensitive competitive enzyme immunoassay (EIA) for measuring SCH 32088 in unextracted human plasma has been developed. The 3-(O-carboxymethyl)oxime (CMO) of SCH 32088 was synthesized and conjugated with bovine thyroglobulin, and the product was used for the production of antibodies in rabbits. SCH 32088-3-CMO was also conjugated with horseradish peroxidase, which was used as the tracer. The EIA thus developed can detect 1 pg SCH 32088 per assay or 25 pg per ml of human plasma. It can reliably quantitate SCH 32088 in human plasma from 50 pg ml-1 to 2.5 ng ml-1 with good linearity, accuracy and precision. The assay can be extended to measure SCH 32088 in plasma of laboratory animals. The availability of this sensitive assay makes it possible to evaluate the pharmacokinetics and toxicokinetics of SCH 32088 in laboratory animals and man.


Use of derivatization reactions with adsorptive stripping voltammetry for determining fotemustine in biological samples. A method is described for determining the new fotemustine antineoplastic in human serum. The method is based on the derivatization of the original molecule by means of diazotization and coupling reactions. The derivatization product is determined by means of adsorptive stripping voltammetry. The calibration graphs were linear over the range 6 x 10(-9)-8 x 10(-7) and 6 x 10(-10)-8 x 10(-8) M, according to whether the coupling reagent was 1-naphthylamine or 1-naphthol, respectively. At the same time, the detection limits are 4.1 x 10(-9) and 1.4 x 10(-10) M, respectively. The method was applied to determine this antineoplastic in human serum, after a liquid-liquid extraction process, from which recovery factors of 95.6% has been obtained.


Fundamental studies in reversed-phase liquid-solid extraction of basic drugs; III: Sample matrix effects. Using a set of 13 basic solutes (including a quarternary ammonium compound) with controlled pKa, log P and plasma protein binding, the factors which influence the extraction of these compounds from plasma by reversed-phase liquid-solid extraction have been studied. Direct analysis of plasma, avoiding deproteination, etc., is possible providing the sample is applied to the cartridge under the appropriate conditions. These conditions are dictated by the degree of plasma protein binding and are controlled and predicted from the lipophilicity of the compound. Plasma samples containing lipophilic compounds (log D(pH 7.4) > 2.03) require acidification and samples containing more polar compounds (log D(pH 7.4) < 0.9) should be applied untreated. Failure to use the appropriate conditions can result in significant losses from the cartridge on application or washing. In the intermediate lipophilicity range the optimal application conditions cannot be predicted. The effect of various aqueous-organic wash solvents have been investigated, and it was found that the cationic but not the proteinaceous components of plasma also affect the extraction process. This phenomenon which is probably due to an attenuation of the secondary cation exchange mechanism, results in more facile elution than when compounds are applied in water.


Relationships between the chromatographic retention data and the effects of nucleoside derivatives in highly metastatic 3LL cells. The effect of 21 nucleoside derivatives on the [3H]-thymidine cellular uptake and on the incorporation into DNA of highly metastatic 3LL (Lewis lung carcinoma) cells has been measured. Hydrophobic and hydrophilic molecular parameters (the adsorption capacity, specific adsorption surface, lipophilicity and specific hydrophobic surface area) have been determined by using thin-layer chromatography. Stepwise linear regression analysis and principal component analysis have been applied in order to reveal the relationships between the molecular parameters and the effect of the nucleoside derivatives on highly metastatic 3LL cells. The first principal component obtained from the measured activity data could be attributed to the change of [3H]-thymidine cellular uptake caused by the nucleoside, while the second principal component could be regarded as the measure of the effect on the DNA incorporation of [3H]-thymidine. The effect of the nucleosides on the [3H]-thymidine uptake could be explained by the specific hydrophobic and adsorption surface area of the nucleoside, on the other hand the effect on the DNA incorporation could be described by the adsorption characteristics (specific hydrophilic surface area and adsorption capacity) of the derivatives.


The enantiomeric resolution of ciprofibrate and related compounds by HPLC using chiral stationary phases. The attempted chromatographic resolution of the drug ciprofibrate and a range of related compounds, containing the chiral moiety 2,2-dichlorocyclopropylbenzene, using five different chiral stationary phases is described. Aqueous mobile phases were used throughout and the successful separation of nine out of 12 pairs of enantiomers was achieved. Structures remote from the chiral centre were seen to affect chiral recognition.


Ion-pair high-performance liquid chromatographic determination of morphine and pseudomorphine in injections. A stability-indicating HPLC procedure has been developed for the determination of morphine and its degradation product pseudomorphine in injection solutions. The reversed-phase chromatographic procedure uses an octadecyl-bonded column with a mobile phase of acetonitrile and aqueous buffer containing an ion-pair reagent. Chromatographic conditions are identified that enable the complete separation of morphine and pseudomorphine without peak deformation or splitting. The only sample preparation required is a simple dilution. The method is found to be precise and linear. The devised method has been applied to the determination of the content of morphine and pseudomorphine in injection solutions stored under normal conditions and after exposure to daylight or heating at 60 degrees C.


Determination of glutathione isopropyl ester in rat, dog and human blood by high-performance liquid chromatography with fluorescence detection. A HPLC method is developed for the determination of glutathione isopropyl ester, a drug for the treatment of cerebral vascular disease, in rat, dog and human blood. The blood is deproteinized with sulphosalicylic acid and the clear supernatant treated with a thiol-specific fluorogenic reagent, ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate (SBD-F) in borate buffer pH 7.5 at 30 degrees C. The derivatization of glutathione isopropyl ester with SBD-F is markedly enhanced by the addition of dimethyl sulphoxide, and is complete in 30 min. The fluorescent derivatives of glutathione isopropyl ester and the internal standard, glutathione ethyl ester are separated from those of endogenous thiols such as cysteine and glutathione on a reversed-phase column. The method is simple and selective with a detection limit of 0.05 micrograms ml-1. Blood concentrations of glutathione isopropyl ester in rats, dogs and humans after intravenous administration are determined using the method.


[A methodology for clinical trials in parenteral nutrition]. Recent and progressive knowledge on parenteral nutrition has not been accompanied by due attention to the methodology for clinical testing in this therapeutical field. Although many different laws, guidelines or norms do exist which can be applied to any clinical test, in the case of parenteral nutrition several characteristic aspects arise that should be taken into account in the design of clinical tests. This review places special emphasis on the definition of clinical testing, the role of the researcher and the ethical, methodological and design-related requirements to be considered when preparing a protocol for study. Not only is the importance of the selection and the size of the sample mentioned, but also the different variables normally used in parenteral nutrition. The correct follow-up, measurement, evaluation and statistical analysis of these variables makes it possible to reach valid conclusions, and provide the basic information for later study. The application of ethical norms and the control of adverse conditions guarantees the correct undertaking of this type of clinical research.


Small solute clearances and clinical outcomes in CAPD. The strong correlation of weekly clearances of urea and creatinine with PCR suggests that minimum targets of 50 L/week/1.73 m2 body surface area of creatinine clearance and a weekly kt/v urea of 1.7 are reasonable if protein intakes in excess of 0.8 g/kg normalized body weight are to be achieved in most CAPD patients. It would seem reasonable that this increases the chances for better nutrition. It would also seem reasonable that this should eventually improve long-term outcomes and survival in large population studies where the impact of dialysis dose could emerge as significant, even with the presence of all the other variables that are known to affect commonly monitored outcome parameters. These recommended targets are well below those achieved by hemodialysis; higher targets could possibly achieve even better long-term results, but this is unknown.


[The methodology of quality control in parenteral nutrition. II. The application of the program to a therapeutic plan]. Patients subjected to parenteral nutrition (PN) must be treated based on a therapeutical plan which has been adapted to their nutritional and clinical needs within a normalizing framework. Furthermore, for this therapeutical plan to be effective and safe, not only must it be suitable for the patient, but also "exact" with regard to daily routines. The quality control programme designed was applied retrospectively to 148 patients. Evaluation was made based on the following indicators: a) delay in initiating parenteral nutrition, b) average intake of macronutrients, c) administration of micronutrients, d) duration of the PN, e) temporary interruptions of the PN, f) errors in medication and g) return of nutrient units. The compliance percentage obtained was 100, 100, 28.7, 89.5, 63.2 100 and 100 respectively. The results obtained have made it possible to understand the quality profile of the therapeutical plan applied to patients on PN. It is also a useful method for the continued improvement of this therapy.


[The Iberian pig: an animal with fat with a high content of monounsaturated fatty acids]. We have analyzed the fatty acid composition of subcutaneous fat in the Iberian pork feed with acorns using gas chromatography. We have found a 59.1% content in oleic acid (18:1) and 62% of total monounsaturated fatty acids. This figures statistically does differs from the data about fat composition in pig taken from others food composition sources. We conclude that Iberian pork feed with acorns have a very high content in monounsaturated fatty acids and can no be considered as harmful as other animals fats.


[Changes in the biochemical parameters during parenteral nutrition. The experience in Hospital de Bellvitge]. The purpose of this study is to evaluate the modifications in biochemical parameters before and after the initiation of nutritional therapy, and to observe whether there is a relationship between the patient's development (exitus or improvement) and the presence of sepsis. The study was performed on 578 adults treated in our hospital from January 1988 to October 1989. The parameters analyzed were the following: glucose, triglycerides, total proteins, albumin, cholesterol, alkaline phosphatase, GOT, GPT, bilirubin, GGT, urea, urates, creatinine and electrolytes. The average initial values of each parameter were compared against those obtained after interrupting the PN by means of the Student t test. The results showed that within the parameters indicating the hepatic function, GGT and alkaline phosphatase were those that showed the most significant differences after ceasing the PN. Furthermore, the parameters indicating hepatic function and the electrolytes showed greater variations, regardless of the clinical evolution of the patient (improvement or exitus). The remainder of the parameters showed significant variations based on the clinical evolution.


Is total creatinine clearance a good predictor of clinical outcomes in continuous ambulatory peritoneal dialysis? The measurement of the adequacy of dialysis in continuous ambulatory peritoneal dialysis (CAPD) is controversial. The use of weekly total creatinine clearance (TCC) has been recommended, but not validated. We analyzed data from our recent urea kinetics in a CAPD study to investigate TCC and its relationship to patient outcomes. TCC was measured over 24 hours by adding residual renal and peritoneal creatinine clearance, correcting for 1.73 m2 surface area and converting to a weekly value. Seventy-six patients had 218 measurements, on starting CAPD and then at 6-month intervals, with mean follow-up of 20 months (range 1-57 months). The mean TCC was 73.62 +/- 32.11 L/week. Due mainly to the loss of residual renal function, the TCC decreased with time (r = -0.40, p < 0.0001), from 88.65 L/week initially to 66.11 at one year, 59.84 at two years, and 50.47 at three years. Dialysate-to-plasma creatinine concentration ratios (D/P Cr) increased with time (r = 0.28, p < 0.0001) from 0.62 initially to 0.66 at one year and 0.73 at two years. The TCC correlated significantly with serum levels of creatinine (r = -0.46, p < 0.0001), urea (r = -0.21, p < 0.001), potassium (r = 0.14, p < 0.05), phosphate (r = 0.25, p < 0.001), and hemoglobin (r = 0.16, p < 0.01), but not with serum albumin or with clinical outcomes including technique failure, hospital days, transfusions, peritonitis rate, nerve conduction velocity, or subjective indices of well-being, except for a weak correlation with the fatigue index (r = 0.19, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


[Percutaneous and surgical gastrostomies: our clinical experience in enteral nutrition]. Percutaneous gastrostomy (PG) was performed on 18 patients and 11 patients were subjected to surgical gastrostomy (SG) in the HCU of Zaragoza from October 1990 to November 1991. Involvement of the high digestive tract was the most common finding: 11 patients had CA of the oesophagus and stomach, 2 suffered from extrinsic pressure of the oesophagus due to metastatic adenopathy of a lung and breast tumour and 9 patients had CA of the larynx with total dysphagia. Another less common indication in our series, although still of great importance, was a severe involvement of the CMS, which made oral nutrition impossible due to functional dysphagia. Due to the long-term nature of this pathology, feeding by nasogastric tube was not considered. This was the case of 4 patients. The duration of enteral nutritional in the patients' home was 112.8 +/- 61 days with PG and 37 +/- 26 days with SG (p: 0.007). The incidence of complications was 33% in PG and 73% in SG, and for both techniques, the most common complication was blockage of the tube. Percutaneous gastrostomy is a simple technique, with no surgical risks involved, but with the benefits of traditional surgical techniques which, despite the small number of cases recorded, has very few complications both with regard to placing and afterwards, and also enables the patient to receive nutrients earlier than would be the case with traditional surgical techniques during prolonged periods at the patient's home.


[Enteral nutrition in burn patients].

[Enteral nutrition in burn patients]. Nutritional support plays an important role in the treatment of patients with burns. Due to the severe hypercatabolism that develops in these patients, oral support is insufficient in most cases, and this makes it essential to initiate artificial nutritional support (either enteral or parenteral). Enteral nutrition is more physiological than parenteral, and data exist which show that in patients with burns, enteral nutrition exercises a protective effect on the intestine and may even reduce the hypermetabolic response in these patients. The purpose of the study was to evaluate the effectiveness and tolerance of enteral nutritional support with a hypercaloric, hyperproteic diet with a high content of branched amino acids in the nutritional support of patients suffering from burns. The study included 12 patients (8 males and 4 females), admitted to the Burns Unit. Average age was 35 +/- 17 years (range: 21-85 years). The percentage of body surface affected by the burns was 10% in two cases, between 10-30% in three cases, between 30-50% in five cases and over 50% in two cases. Initiation of the enteral nutrition was between twenty-four hours and seven days after the burn. The patients were kept in the unit until they were discharged, and the average time spent in the unit was 31.5 days (range: 17-63 days). Total energetic requirements were calculated based on Harris-Benedict, with a variable aggression factor depending on the body surface burned, which varied from 2,000 and 4,000 cal day. Nitrogenous balance was determined on a daily basis, and plasmatic levels of total proteins, albumin and prealbumin on a weekly basis. There was a significant difference between the prealbumin values at the initiation and finalization of the enteral nutrition (9.6 +/- 2.24 mg/dl compared with 19.75 +/- 5.48 mg/dl; p < 0.001). The nitrogenous balance improved, changing from -5.4 in the second week to positive values by the fourth and fifth weeks of treatment. Tolerance to the enteral diet was very good, and only mild complications such as diarrhoea developed in two patients. Enteral nutrition is a suitable nutritional support method for patients with burns, which maintains the nitrogenous balance positive and improves the visceral protein parameters in these patients at an early stage, with very few complications.


Intraperitoneal administration of phosphatidylcholine improves ultrafiltration in continuous ambulatory peritoneal dialysis patients. Reports in the literature have linked a low phosphatidylcholine content in continuous ambulatory peritoneal dialysis (CAPD) effluent to ultrafiltration loss. Clinical evidence suggests that adding phosphatidylcholine to the dialysis solution enhances ultrafiltration. A clinical study has been designed to clarify the effect of phosphatidylcholine on ultrafiltration in CAPD patients with normal ultrafiltration. A weekly measurement of the peritoneal equilibration test was conducted per patient in the hospital. A comparison between the control dialysis solution (three-week period) and the phosphatidylcholine premixed solution (three-week period) was performed on a total of 12 patients. This study shows that a phosphatidylcholine premixed dialysis solution significantly enhances ultrafiltration. Since ultrafiltration per osmotic driving force (mL/g glucose) is enhanced, the patient's glucose load per day is reduced to achieve equal ultrafiltration. In the presence of phosphatidylcholine, peritoneal permeability remained unchanged, as indicated by membrane transport characteristics. No side effects were observed.


Dialysis at home in the west of Scotland: a comparison of hemodialysis and continuous ambulatory peritoneal dialysis in age- and sex-matched controls. To audit the outcome of patients treated at home by hemodialysis and continuous ambulatory peritoneal dialysis (CAPD). Retrospective comparison of nondiabetic hemodialysis patients with age- and sex-matched nondiabetic patients treated by CAPD. Renal Units, Stobhill General Hospital and Western Infirmary, Glasgow, providing the home dialysis service for the West of Scotland. Between 1982 and 1988, 139 hemodialysis patients starting treatment at home, compared with 139 matched patients starting CAPD over the same time period. Patient characteristics and cardiovascular risk factors at the start of home treatment. Patient and technique survival with both forms of dialysis. Patients selected for home hemodialysis were less likely to be smokers (p < 0.02) and to have electrocardiographic evidence of ischemia or left ventricular hypertrophy (p < 0.05) than patients treated by CAPD. Patient survival and technique survival (excluding death and renal transplantation) at 3 years were 93.8% versus 86.2% (p < 0.05) and 94.2% versus 80.8% (p < 0.04) for hemodialysis and CAPD, respectively. Cardiovascular events were responsible for the majority of deaths in both groups, but there was a greater proportion of deaths from other causes in patients treated by CAPD. There was no significant difference in the transplantation rate between the two treatment groups. Home dialysis is an effective method of renal replacement treatment for patients with end-stage renal disease. The results of hemodialysis are superior to CAPD, but this may be partly due to selection bias.


The pharmacokinetics of intraperitoneal erythropoietin administered undiluted or diluted in dialysate. To compare the bioavailability of intraperitoneal erythropoietin (EPO) administered undiluted versus diluted in 2 L of dialysis fluid. Group 1 patients received one dose of EPO, 400 U/kg BW given with vehicle only. This dwelled for 8 hours after which 2 L of dialysate were infused. Group 2 patients received the same dose of EPO diluted in 2 L of dialysate which dwelled for 8 hours. Both groups resumed their CAPD regimen after the first 8 hours. Blood levels of EPO were measured for 24 hours in both groups. The Home Peritoneal Dialysis Unit, Toronto Hospital, Western Division. The participants were on CAPD for at least three months, free of peritonitis, and had no abnormalities of peritoneal transport. Three patients took part in both arms of the study, and there were 6 patients altogether in each group. When EPO was administered undiluted, there was a greater than ninefold increase in bioavailability of the hormone as measured by the area under the curve (AUC), compared to when the same dose was diluted in 2 L of dialysis fluid. The previous studies that reported low bioavailability of intraperitoneal EPO used the hormone diluted in dialysate. The current findings suggest that if EPO is given in the dry peritoneal cavity, the bioavailability is greatly improved and may be clinically effective. Intraperitoneal instillation may prove to be an alternative route for EPO in the peritoneal dialysis patient unable or unwilling to receive subcutaneous injections. We are currently studying the effectiveness of undiluted intraperitoneal EPO in CAPD patients.


Effect of erythropoietin in continuous ambulatory peritoneal dialysis patients: comparison between intravenous and intraperitoneal administration. The administration of recombinant human erythropoietin (rHuEPO) in CAPD patients is usually done subcutaneously. Only a few authors have reported on its intraperitoneal (IP) administration. We compared the effect of IP administration of rHuEPO in CAPD patients to that of intravenous (IV) administration. Ten anemic CAPD patients injected rHuEPO into their dialysis bag once a day, 3 times a week, for 18 weeks. The initial dose was 12,000 U. The dwell time of the exchanges with rHuEPO was about 6 hours. Nine other anemic CAPD patients were treated with IV rHuEPO once a week for 18 weeks. The initial dose was 6000 U. In the IP group the hematocrit rose from 24.04 +/- 2.7% to 33.3 +/- 3.8% (mean +/- SD). In the IV group 2 patients were excluded from the efficiency evaluation. In 7 of the 9 patients in the IV group, the hematocrit rose from 23.27 +/- 2.6% to 32 +/- 5.5% (mean +/- SD). The intraperitoneal administration of rHuEPO in CAPD patients is sufficient in improving anemia, although it requires a much larger dosage to yield the same level of improvement as the one obtained with the intravenous administration. However, in patients on continuous cycling peritoneal dialysis or IP dialysis, a smaller dosage during the prolonged dwell time may be effective.


Intraperitoneal administration of recombinant human erythropoietin. To determine the efficacy and safety of intraperitoneal administration of recombinant human erythropoietin (rHuEPO) in continuous ambulatory peritoneal dialysis (CAPD) patients compared to subcutaneous rHuEPO. Prospective analysis of an open, nonrandomized investigation. Outpatient CAPD clinics in two university hospitals. Nine adult CAPD patients receiving rHuEPO intraperitoneally and 8 patients receiving rHuEPO subcutaneously. One hundred units of rHuEPO per kilogram of body weight were administered three times a week for 8 weeks or until the target hematocrit of 35% was reached. Thereafter, dosages of rHuEPO were adjusted for response. Intraperitoneal rHuEPO was administered in 1 L of dialysis solution during the night. Efficacy was assessed by measuring the increase in hemoglobin. Tolerance was assessed by monitoring side effects. In the first 8 weeks of treatment hemoglobin concentration increased from 64.5 +/- 12.9 g/L to 98.3 +/- 16.1 g/L (p < 0.0005) in the intraperitoneally treated group. In the subcutaneously treated group hemoglobin increased significantly faster (p < 0.05) from 72.5 +/- 4.8 g/L to 119.2 +/- 11.3 g/L (p < 0.0005) in the same period. Antihypertensive medication had to be increased or instituted in most of the patients in both groups. The incidence of peritonitis in the intraperitoneally treated group was not increased when compared to the pretreatment incidence. Subcutaneously administered rHuEPO is superior to intraperitoneally administered rHuEPO with regard to the required dosages. However, the results of this study show that intraperitoneal administration of rHuEPO might be a convenient and safe alternative when subcutaneous administration is undesirable.


Six-year experience with swan neck catheters. From the beginning of our continuous ambulatory peritoneal dialysis (CAPD) program in January 1977 until June 1985, we used Tenckhoff and Toronto Western Hospital catheters. Throughout these years catheter survival probabilities of about 30% at three years persisted unchanged and were similar to survival probabilities reported by the National CAPD Registry special survey for these catheters. The first improvement in catheter results regarding leaks was noted after the adoption of lateral catheter insertion. Malfunction was less using swan neck prototypes from August 1985 to April 1986. The latter catheters were made of 80 degrees arc angle tubing between 8.5 cm spaced cuffs and were inserted in a reversed U-shape tunnel with the incision at the top of the tunnel. The use of these catheters was abandoned because of high cuff extrusion and exit infection rates. The next generation of swan neck catheters, the swan neck Missouri 2 and 3 catheters with straight intraperitoneal segments, improved the results dramatically. These catheters were made of 180 degrees arc angle tubing between 5 or 3 cm spaced cuffs. The estimated survival probability of 61% at three years more than doubled compared to previously used catheters. Recently we modified the intraperitoneal segment of the catheters, replacing the straight segment with a coiled one. These modified catheters, the swan neck Missouri coiled catheters, have been used exclusively since February 1990. In addition to an acceptable survival probability of 88% at one year, there are two major advantages of these catheters, the same as for other coiled catheters: elimination of infusion pain due to a jet effect and pain related to straight catheter tip pressure on the peritoneum experienced by some patients.


The evaluation of low-dose cytarabine in the treatment of myelodysplastic syndromes: a phase-III intergroup study. One hundred and forty one patients were treated in a combined Eastern Cooperative Oncology Group and Southwest Oncology Group phase-III study evaluating low-dose cytarabine (LDAC) versus supportive therapy for the treatment of myelodysplastic syndrome (MDS). Patients were randomized to either cytarabine 10 mg/m2 subcutaneously BID or supportive therapy. Central pathology review was required. All patients were classified according to the FAB criteria for MDS. The overall concordance rate for the MDS subtype was 52%, and 25 patients were pathology exclusions, including 20 with AML. The overall response rate to a single cycle of LDAC was 32%, with 11% complete and 21% partial responses. The median duration of response was 5.9 months, with a range of 1.4-33.5 months. Responses were seen in all subtypes. Infections were more common in the LDAC arm. There was no difference in the time to progression or the overall survival for patients treated with LDAC or supportive therapy. The incidence of leukemic transformation was similar in both arms at 15%, but it differed according to the MDS subtype. Patients receiving LDAC had a decreased transfusion requirement after 3 months. There was a significant correlation between the degree of cytoreduction after receiving a single cycle of LDAC and survival. This survival difference was most marked in patients with the RAEB and RAEB-T subtypes. Although LDAC produced responses in all subtypes of the MDS, there was no effect on overall survival or transformation to AML. However, selected patients benefited from a single cycle of LDAC with durable responses. A cytoreductive effect appears to be required for a durable response. Future studies should include pathology review and must address the clinical and biological heterogeneity of MDS.


Flow-cytometric enumeration of reticulocytes with the new fluorochrome 1,3'-diethyl-4,2'-quinolylthiacyanine (correction of 1',3'-diethyl-4,2'-quinolylthiacyanine). Several flow-cytometric methods for reticulocyte enumeration in whole blood have been developed, with different degrees of practical use. Recently, a new fluorochrome, 1,3'-diethyl-4,2'-quinolylthiacyanine (corrected) (DEQTC) was proposed in a brief report, as an alternative to thiazole orange for reticulocyte counting. We have evaluated the usefulness of this fluorescent stain by assessing the optimal conditions for the flow-cytometric analysis, and by comparing in double-blind assays the quantitative results of this technique with those obtained by manual counting with brilliant cresyl blue. Our results show that flow cytometry with DEQTC is highly correlated to the manual method (r = 0.95-0.99), supporting the interest of this particular stain and of flow cytometry for routine laboratory work in hematology.


A major concanavalin-A-binding cell surface protein from normal and leukemic granulocytes: isolation and characterization. This paper reports the isolation and biochemical characterization of a major concanavalin A (Con A)-binding cell surface protein (protein 2, M(r) 75-85 kD) from normal and chronic myeloid leukemic (CML) granulocytes. Our studies show that protein 2 has two differentially glycosylated forms, protein 2a (M(r) 75-85 kD), which does not bind the lectin RCA, and protein 2b (M(r) 80-90 kD), which does. Both molecules show identical retention times on reverse-phase HPLC, irrespective of the cell source. By the procedure used the amount of 2a obtained is about 2.4 times more than that of 2b in normal cells and about 2.6 times more in CML cells. Furthermore, both are approximately 2.4-fold more in CML granulocytes. A polyclonal antibody to protein 2a also immunostains protein 2b. The antibody to protein 2a does not prevent Con A binding but inhibits its internalization. Similarity of the molecules from both the cell types and their increased amounts in CML granulocytes suggest that factors/components other than its structure and amount are responsible for the known defective internalization of Con A by CML granulocytes.


Rapid molecular characterization of mutations leading to unstable hemoglobin beta-chain variants. Characterization of unstable hemoglobins by protein analysis is often difficult. However, it is facilitated by DNA analysis, especially in the case of hyperunstable beta-chain variants, which produce a beta-thalassemia phenotype. We have applied an efficient strategy to the detection of such variants at the DNA level, based on computer-designed denaturing gradient gel electrophoresis (DGGE) of amplified DNA fragments. This approach makes it possible to detect any anomaly in the beta-globin gene. We describe the use of the DGGE method for rapid characterization of beta-chain variants and report a new missense mutation in the beta-globin gene third exon, beta 127 CAG-CGG/Gln-Arg, which is responsible for the synthesis of a highly unstable hemoglobin.


A definition of bias founded on the concept of the study base. We propose a definition of bias founded on the concept of the study base. The definition is unrelated to the study design, thereby permitting a clear comparison of validity among different study types. Also, it facilitates assessment of bias because it allows bias to be subdivided under mutually exclusive categories. In relation to the study base, the epidemiologic study can be viewed as involving three distinct steps. First, the person-time (study base) is specified for the study. Second, the investigators collect data on disease events and person-time among the exposed and unexposed from the study base. Third, they calculate an estimate of the effect of the exposure using the data. Categories pertaining to each of these three steps will create mutually exclusive categories of bias. We suggest the following terminology: the first step generates confounding, the second results in misclassification and misrepresentation, and the third produces analysis deviation. The total bias is the sum of the confounding, misclassification, misrepresentation, and analysis deviation. We illustrate our proposed epidemiologic theory with examples from various biological settings.


Renal cell cancer among paperboard printing workers. A physician's alert prompted us to investigate workers' cancer risk at a paperboard printing manufacturer. We conducted a retrospective cohort mortality study of all 2,050 persons who had worked at the facility for more than 1 day, calculated standardized incidence ratios (SIRs) for bladder and renal cell cancer, and conducted a nested case-control study for renal cell cancer. Standardized mortality ratios (SMRs) from all causes [SMR = 1.0, 95% confidence interval (CI) = 0.9-1.2] and all cancers (SMR = 0.6, 95% CI = 0.3-1.0) were not greater than expected. One bladder cancer and one renal cell cancer were included in the mortality analysis. Six incident renal cell cancers were observed, however, compared with less than two renal cell cancers expected (SIR = 3.7, 95% CI = 1.4-8.1). Based on a nested case-control analysis, the risk of renal cell cancer was associated with overall length of employment but was not limited to any single department or work process. Although pigments containing congeners of dichlorobenzidine and o-toluidine had been used at the plant, environmental sampling could not confirm any current exposure. Several limitations and a potential selection bias limit the inferences that can be drawn.


Evaluation of three nutritional assessment methods in a group of women. We conducted a reproducibility study of four 24-hour dietary recalls (N = 224) and four biochemical assessments of nutritional status (N = 265) in a group of women in Alabama. For 24-hour recalls, the variance component ratios were all greater than 1, and the intraclass correlation coefficients ranged from 0.16 to 0.27 for macronutrients, and from 0.09 to 0.37 for vitamins and minerals. The intraclass correlation coefficients for biochemical assessments ranged between 0.39 and 0.74 with corresponding variance component ratios of 1 or below for most nutrients. The correlation coefficients between the food frequency questionnaire on the usual dietary intake during the year preceding the beginning of study and the mean values of four 24-hour dietary recalls administered at the initial visit and again after 2, 4, and 6 months ranged from 0.3 to 0.4 for most nutrients. We found plasma beta-carotene levels to be moderately correlated with dietary vitamin A (r = 0.20) and beta-carotene (r = 0.22).


Assessing consumption of high-fat foods: the effect of grouping foods into single questions. Questionnaires on the frequency of consumption of foods are commonly used to measure dietary intake in epidemiologic research. To reduce the burden on respondents, questionnaires are often shortened by combining inquiries on similar foods into a single question. The effect of this practice on the reporting of dietary intake has never been investigated, however. To address this issue, we used two food frequency questionnaires in a telephone survey designed to rank adult residents of Alabama by their intake of dietary fat. One questionnaire included 29 questions about separate high-fat foods, whereas the other grouped these same foods into 14 questions. Compared with the 443 respondents interviewed using the 29-item separated-foods questionnaire, the 465 respondents responding to the 14-item grouped-foods questionnaire reported lower average intakes of the foods. In addition, a substantially higher percentage of respondents to the grouped-foods questionnaire reported never consuming the foods.


Passive smoking in Germany and Poland: comparison of exposure levels, sources of exposure, validity, and perception. Nonsmoking females age 35-65 years from Bremen, Germany (91 women), and Opole, Poland (98 women), were interviewed about their recent passive smoking exposure. We obtained urine samples at the time of interview and determined the concentration of cotinine as an indicator of tobacco smoke exposure. In Poland and in Germany, the vast majority of nonsmoking women are involuntarily exposed to environmental tobacco smoke (ETS). Polish women had slightly higher exposure levels than German women, with overall means of 9.93 and 8.65 ng cotinine/mg creatinine, respectively. Smoking by the husband was the major source of exposure in both study groups. In the Polish group, the work place was also an important source of ETS exposure. The validity of self-reported passive smoking exposure was found to be generally good; it was somewhat better in the German study group. A negative attitude toward tobacco smoke was slightly stronger among the German women. The percentage of women misreporting their active smoking status was low.


Hospitalization with vertebral fracture among the aged: a national population-based study, 1986-1989. Whereas fractures related to osteoporosis have become a pressing public health concern, relatively few epidemiologic studies have focused on vertebral fractures. To shed further light on the occurrence of this injury, we collected data from the Health Care Financing Administration on 151,986 discharges listing a diagnosis of vertebral fracture over a 4-year period. After adjusting for age, white women experienced the highest rates of discharge, at 17.1 per 10,000 per year, followed by white men (9.9 per 10,000), black women (3.7 per 10,000), and black men (2.5 per 10,000). Among white women, discharge rates rose exponentially from 5.3 discharges per 10,000 population at age 65 to nearly 47.8 per 10,000 at age 90. White men, black women, and black men experienced less dramatic age-related increases in discharge rates. The similarity of these patterns to discharge rates for hip fracture suggests that the race-sex differences in vertebral fracture discharge rates may be due to differences in the incidence of vertebral fracture.


A case-control study of smoking and gallbladder disease: importance of examining time relations. We report a case-control study of gallbladder disease and cigarette smoking. Subjects were 252 incident cases and 233 age-sex-residence-matched community controls. Three quarters of the subjects were women. The age-matched risk of gallbladder disease was increased in current smokers vs never-smokers [odds ratio (OR) = 1.6 and 1.3 in women and men, respectively] and in past smokers (OR = 1.5 for each sex). In light of earlier research, showing that, in women, certain risk factors for gallbladder disease may act maximally soon after first exposure, we examined time relations. In women, multivariable analysis showed that the risk increase was especially high in current-smoking women less than 35 years of age [OR = 3.5, 95% confidence interval (CI) = 1.2-9.8] and in women who had been smoking for 1-8 years (OR = 2.8, 95% CI = 1.1-7.1). These results indicate that, in women, the smoking-related risk of gallbladder disease is greatest soon after first exposure--as has also been reported for oral contraceptive use. The findings highlight the potential for mis-estimating risk in case-control studies when late-occurring cases have a different relation to the exposure factor than do the early-occurring cases within the original, albeit undocumented, exposure cohort.


Mammary gland mass and breast cancer risk. Evidence indicates that early life events and conditions, possibly extending to the intrauterine stages of life, and including energy restriction in early life, affect the risk of breast cancer. The mechanism of this effect is likely to be through a reduction in mammary gland mass and, inferentially, the total number of ductal stem cells. The evidence derives from epidemiologic and animal studies. It can explain certain epidemiologic findings that cannot be accounted for by more established breast cancer risk factors, including the more frequent occurrence of breast cancer in the left breast and the higher incidence of this disease among caucasian women than among Asian women in Asia.


Lactation history and bone mineral density among perimenopausal women. To examine the relation between lactation and bone mineral density, we conducted a cross-sectional study among white, nonsmoking, perimenopausal women age 40-54. Three hundred fifty-two women completed a questionnaire covering medical and reproductive histories, physical activity, and diet. We measured the bone density of lumbar vertebrae 2-4 with dual photon absorptiometry, and the midradius and distal radius with single photon absorptiometry. Women with a history of lactation had 41 mg per cm2 higher lumbar bone mineral density than women with no lactation (95% confidence interval = 8-74 mg per cm2), controlling for parity, body mass, physical activity, and menopausal status. We found no important increase at mid- or distal radius.


Reliability of sexual behavior data among high school students in Rome. We compared findings on sexual behavior from a self-administered questionnaire and a face-to-face interview, separated by 2-4 weeks, directed at 383 Italian secondary school students. The correlation coefficients or kappa values for reported prevalence of coital experience, age at first intercourse, and age of first partner were all greater than 0.7. The interview underreported coitus and overreported condom use, when administered before the questionnaire. The students thought that 18.4% of their classmates had ever had coitus, compared with 23.5% on self-report. We conclude that despite an overall high reproducibility of sexual behavior measures, distortions due to perceptions of social desirability might occur.


The reliability of self-reported alcohol consumption in the remote past. We examined the reliability of self-reported alcohol consumption in past age periods of women's lives. As part of a case-control study of breast cancer conducted in Massachusetts and Wisconsin in 1988-1991, the same questionnaire was administered for a second time to 211 controls (mean age = 54 years) after an interval of 6-12 months. The Spearman correlation coefficients between the average number of grams of alcohol consumed daily reported in the two interviews, by age period of consumption, were: 16-19 years, r = 0.81; 20-29 years, r = 0.84; 30-39 years, r = 0.75; and for recent consumption, r = 0.77. Self-reported alcohol consumption throughout adult life was reported with precision sufficient to make the ranking of subjects' intake consistent between interviews.


A research-based risk assessment tool as the cornerstone of a pressure ulcer prevention program. By highlighting theories of knowledge utilization, we consider some of the factors and issues in planning research utilization projects. By our use of a research utilization model in developing a research-based pressure ulcer prevention program, we help to bridge the gap between research and practice.


Fibronectin in acute and chronic wounds. Fibronectin, a fibrillar glycoprotein, is the major adhesive protein deposited at the wound interface during acute wound healing. Its abilities to interact with both cells and matrix are responsible for its description and characterization as a multifunctional adhesive glycoprotein. Fibronectin functions to promote cell adhesion, cell shape and spreading, organization of the cytoskeleton, hemostasis and thrombosis, cell migration, and phagocytosis. Both in vitro and in vivo studies indicate that fibronectin interacts with cells important to wound healing, including platelets, neutrophils, monocytes, fibroblasts, endothelial cells, and epidermal cells. It promotes adhesion between cells and between cell and matrix, cell spreading, migration, chemotaxis, phagocytosis, and matrix and basement membrane organization. Fibronectin is consequently active during each stage of wound healing. Its importance in acute wound healing is now widely recognized. More recently, investigators have begun to study fibronectin's role in chronic wounds. This article briefly reviews fibronectin, outlines its role in wound healing, and discusses current research on fibronectin in chronic wounds.


Air support therapy: ethical considerations. Pressure ulcers are a major health problem for the elderly population in the United States. Billion of health care dollars are spent for hospitalization, air support therapy, wound care products, and nursing care. In addition, the cost in human suffering is priceless. Many patients with pressure ulcers have several chronic diseases that make wound healing difficult. Nurses must begin to examine the ethical principles that guide their care of these patients. This article focuses on some ethical issues related to the care of patients with pressure ulcers and the difficulties encountered with pressure ulcer care. Careful analysis of the goals of pressure ulcer treatment may help nurses develop ethical guidelines to direct them in the care of these patients.


Trends in Indian adolescent drug and alcohol use. Trends in overall drug use among Indian and non-Indian youth have followed similar patterns, increasing from 1975 to the early 1980s and, for the most-used drugs, declining since then. At every point in time more reservation Indian youth are involved with drugs than are non-Indian youth. Rates for cocaine and hallucinogen use by Indian youth increase until 1990. The decline in overall drug use has occurred because a considerable number of moderate users have shifted to non-use. There has been no decrease in the proportion of high-risk users; since 1980, it has stayed between 17% and 20%. Societal changes and prevention programs are reaching casual drug users but not those susceptible to heavy drug involvement.


Comparison of drug use rates for reservation Indian, non-reservation Indian and Anglo youth. Rates of drug use and involvement were compared for three groups: Indian youth living on reservations, Indian youth living off reservations and Anglo youth. A consistent pattern emerged, showing the lowest rates of use among Anglo youth, higher rates among non-reservation Indian youth, and the highest rates among Indian youth on reservations. Rates of tobacco use, both smoked and smokeless, and marijuana use are especially high for Indian youth. Indian youth also show a pattern of earlier initiation to drug use. Gender differences reveal slightly higher rates of use for males, although the differences are not great enough to suggest that prevention efforts for males should have a higher priority.


Clinical trial of the Braden Scale on an acute care medical unit. Pressure ulcers in hospitalized patients represent a significant challenge to the health professionals who care for these patients. This challenge extends to the health care system as a whole because of the staggering costs associated with pressure ulcer resolution. Although the relative benefits of various treatment methods have long been debated, intervention before ulcer formation clearly stands as the most efficacious approach to this problem. The Braden Scale for Predicting Pressure Sore Risk, a recently developed paper tool, was evaluated in our acute care medical unit for its ability to predict pressure ulcer occurrence. The sensitivity of the Braden Scale in this evaluation was much lower than that reported in other published studies. The Braden Scale was less effective than the nurse judgment prediction method, the current prevailing prediction method at this institution. The gravity of these results, however, is unclear because of the questionable incidence of ulcers in this study, which may be artificially high due to permissive criteria used in the identification of stage 1 ulcers. We determined that the lack of unequivocally defined criteria for the identification of stage 1 ulcers may contribute to variance between studies. We also found that intervention measures were used only 27% of the time after the identification of patients at risk.


The consequences of drug and alcohol use for Indian youth. Indian youth have higher rates of using alcohol and drugs in ways that increase their risk--getting very drunk, drinking while driving, and using drugs and alcohol together. The highest rate is found among reservation youth, a lower rate among non-reservation Indians, and the lowest rate among non-Indian youth. Frequency of self-reported consequences from alcohol and drug use follow the same order, with 15% of reservation seniors involved in an alcohol-related accident. The most frequent consequences involve relationship problems. Drug injection is rare in all groups.


Attitudes about drugs and the drug use of Indian youth. Alcohol, marijuana, and inhalants are the easiest drugs to obtain, but all drugs are available to some students. Younger students felt that inhalants were easier to get than marijuana. Availability does not have a major effect on use; if there is motivation to use, drugs are available. Perceived harm is linked to use, and 8th-grade reservation youth show the lowest belief that drugs are harmful; only 51% believe that using marijuana regularly will lead to "a lot" of harm. In general, non-Indian youth show higher rates of perceived harm, congruent with their lower rates of drug use.


Drug use of friends: a comparison of reservation and non-reservation Indian youth. There is not much drug use at school, but reservation youth are most likely, non-reservation Indian youth next most likely, and non-Indian youth least likely to have used drugs at school. Alcohol and drugs are used most frequently at weekend parties and at night with friends. Reservation youth are twice as likely as the others to have used alcohol while driving around and three times more likely to have used drugs while driving around. Drugs are used with friends. Heavy drug users are more likely to have friends who use drugs and friends who encourage drug use and are less likely to have friends who would try to stop them from using drugs.


Characteristics of Indian youth and drug use. The overall high rates of drug use found among Indian youth may be accounted for in part by lack of educational and employment opportunity and other endemic problems of Indian reservations. Individual drug involvement is most highly related to membership in drug-using peer clusters; but because of physical isolation, links between drug use and close friends are weaker for Indian youth, and family influence is felt more strongly. Anxiety, depression, and low self-esteem are not related to drug involvement, but angry youth are more likely to have drug-involved peers. Risk factors for Indian youth are low family caring, age first drunk, poor school adjustment, weak family sanctions against drugs, positive attitudes toward alcohol use, risk of school dropout, father not at home, and poor religious identification.


Enzyme histochemistry of the olfactory and vomeronasal sensory epithelia in the golden hamster. Histochemical activities of several enzymes were investigated in the olfactory epithelium (OE) and vomeronasal organ (VNO) of the golden hamster. Activities of adenosine triphosphatase, lactate dehydrogenase and succinate dehydrogenase were intense in the OE, and the sensory (VSE) and respiratory epithelium (VRE) of the VNO. The activity of acid phosphatase was intense in both the OE and the VSE, while that of non-specific esterase was intense in the VSE alone. The activity of alkaline phosphatase was detectable only in the VRE. Activities of monoamine oxidase and acetylcholine esterase were negative in all of the OE, VSE and VRE. These similarities and differences in the histochemical distribution of enzymes between OE and VSE may reflect the common olfactory function and/or functional specialization in these epithelia. On the other hand, the VRE was considerably different from the OE and VSE in the enzymatic distribution. This may reflect the non-olfactory function of this epithelium.


Effects of pulsing electromagnetic fields on the ligament healing in rabbits. Effects of pulsing electromagnetic fields (PEMFs) on ligament healing were investigated using 80 rabbits. All animals received square resection (4 mm x 4 mm) of both patellar ligaments in full thickness at their center. They were divided into 4 groups of 20 rabbits each and stimulated with different electromagnetic intensity of 0 (control), 2, 10, and 50 gauss (G) for 6 hr daily. Pulse frequency and pulse width were 10 Hz and 25 microseconds, respectively. After PEMFs stimulations for 1, 2, 3 and 4 weeks, 5 animals of each group were euthanized and the regenerated tissue at the defective portion was investigated histologically and biomechanically. Histologically, the tissue stimulated by PEMFs showed an earlier increase in capillaries and fibroblasts and more matured, prominent longitudinal orientation of collagen fibers than those of control groups. Among the rabbits stimulated electromagnetically, those stimulated at 50 G revealed the earliest ligament healing. Tensile strength of regenerated ligament tissues of any PEMFs groups increased significantly at 1 and 2 weeks after operation, however, at 3 and 4 weeks after operation, there were no significant differences between groups. Among these values, those of 50 G group were the highest consistently during most of the experimental period. From the above results, PEMFs enhanced the earlier stage of ligament healings and 50 G gauss seemed to be the most effective among the 3 field intensities used. This promoting effect may potentiate the earlier recovery of the function after the ligament injury.


Production and some properties of cytotoxins produced by Campylobacter species isolated from proliferative enteropathy in swine. Cytotoxin production by Campylobacter species isolated from proliferative enteropathy in swine was examined. Twenty-one of 29 strains of C. hyointestinalis, 10 of 27 strains of C. mucosalis and 10 of 10 strains of C. coli were cytotoxin positive. By the gel filtration chromatography of C. hyointestinalis culture filtrate, cytotoxin activities were observed in two peaks (fraction I and fraction II). Most of the cytotoxic activities lay in fraction I, which is heat-labile, trypsin-sensitive and the molecular weight was estimated at 40,000. On the other hand, fraction II cytotoxin was heat-stable, trypsin-insusceptible and molecular weight was approximately 1,000.


Histochemical studies on the dependence of secretory function of the major vestibular gland (Bartholin's gland) on ovarian steroid hormones in the cat. The dependency of the secretory function of the feline major vestibular gland (Bartholin's gland) on the ovarian steroid hormones was studied histochemically. After estrogen (estradiol-17 beta or diethylstilbestrol), progesterone or a combination of these was administered to cats which were previously ovariohysterectomized, the major vestibular glands were removed, embedded in paraffin, sectioned, and stained by alcian blue, periodic acid-Schiff or peroxidase-labeled lectin (peanut agglutinin and wheat germ agglutinin). The vividly positive reactions to the stainings applied were observed in the secretory epithelial cells of the major vestibular glands in the estrogen treated animals. The dependency of the secretory function of the major vestibular glands on the estrogen was demonstrated in this study.


Evaluation of the anaerobic method for the analysis of fecal microflora of beagle dogs. The plate-in-bottle method of Mitsuoka et al. (1969) for the counting of fecal bacteria in beagle dogs was superior to an anaerobic jar method. Comparisons of three nonselective media, such as medium 10 supplemented with 10% cecal extract of dogs (designated as M10C), M10 with 10% fecal extract of dogs (M10F), and M10, by the plate-in-bottle method indicated that the visible bacterial counts for M10C were higher than those for M10 and M10F. The high percentage (18.4%) of numbers of the extremely oxygen-sensitive anaerobes to the fecal total counts by using the plate-in-bottle method with M10C was also observed.


In vitro study on the increase of trypsin resistance in the rat testicular sperm head. The alteration of trypsin resistance in rat sperm head was investigated after the sperm were incubated in the culture media (199-Earele, 1% BSA) containing the inhibitors of protein synthesis. The head of non-incubated testicular sperm was easily digested by trypsin (1 mg/ml) while the head of the incubated sperm showed resistance to trypsin digestion after 3 hr-incubation. Trypsin resistance in epididymal sperm head did not change after the sperm was incubated. Neither cycloheximide (100 micrograms/ml) nor chloramphenicol (100 micrograms/ml) affected the trypsin resistance in the testicular sperm head.


Activities of certain cytoplasmic enzymes in bovine T lymphoma. Activities of certain cytoplasmic enzymes were measured in bovine T lymphoma (BTL-PC3 cells). The activities of hexokinase, pyruvate kinase and glucose-6-phosphate dehydrogenase in PC3 cells were elevated as much as 2 or 3-fold of those in bovine thymocyte. The high activities of these enzymes derived from activation of glucose metabolism may reflect the high growth potential of PC3 cells.


Effect of cobra venom factor on experimental infection of mice against Clostridium chauvoei. The effect of cobra venom factor (CoVF) treatment was examined to clarify the mechanism of resistance of mice to Clostridium chauvoei infection. In CoVF-treated mice inoculated with spores of C. chauvoei, no death occurred and the organisms in the infected muscle progressively decreased, similar to that of non-treated control mice. These results indicated that C3 did not play a significant role in the resistance of mice against C. chauvoei infection.


Surgery of hepatic cysts in a cat.

Surgery of hepatic cysts in a cat. A 2-year-old female Japanese domestic cat weighing 3.6 kg was presented due to marked abdominal distention and was diagnosed as hepatic cyst. The bile acid concentration of the hepatic cyst was as low as the serum bile acid concentration. The membrane of the cysts was excised from the edge of the compressed hepatic tissue and no other surgical manipulation was performed. This surgical procedure is simple and quick compared to hepatic lobectomy, however, it is necessary to evaluate the fluid of the cyst before performing this procedure.


Enhanced resistance to bacterial infections in mice by oral administration of an active egg white product. We found that the oral administration of active egg white product (AEWP) enhances host resistance to infections. The prophylactic administration of AEWP enhanced nonspecific resistance in mice to systemic infection with E. coli. Moreover, the therapeutic administration of AEWP was effective against S. aureus infection in mice.


Comparison of the inhibitory effect of sulfamonomethoxine and other sulfonamides on capsule formation of Bordetella bronchiseptica. The inhibitory effect of sulfamonomethoxine and other sulfonamides on the capsule formation of sulfonamide-resistant Bordetella bronchiseptica was investigated. All the sulfonamides having MeO(-OCH3) groups inhibited the capsule formation of B. bronchiseptica. Strong inhibition was obtained with sulfamonomethoxine. Inhibition was not seen with sulfonamides having no MeO groups.


Visual evoked potentials in guinea pigs with brain lesion. Visual evoked potentials (VEPs) were recorded in 10 adult male guinea pigs with brain lesion. Lesions were produced in 5 animals by superficial suction of the occipital lobe. The other 5 animals were orally administered with hexachlorophene (about 35 mg/kg/day) for 28 days. In the VEP following the ablation of the occipital lobe, the peaks P10, N20, P55, N75, N140 and P200 disappeared in many cases. The amplitude of the peak N40 decreased to approximately one half its control VEP. In the VEP obtained from the animals administered with hexachlorophene, the peak latencies of N20, P30, P55, N75 and P100 were slightly prolonged after the 7th day following the first administration. On the other hand, there was no change in the latency of N40 during the whole period of administration. The peak-to-peak amplitude showed some variability in different peaks. Histologically, diffuse status spongiosis were found in the white matter of the cerebrum, cerebellum, and brain stem. As described above, the ablation of the occipital lobe caused markedly depressed VEPs, however, the responses to the photic stimulation persisted after the injury. On the other hand, the VEPs of animals administered with hexachlorophene showed a high probability of peak appearance, and a decrease in amplitude was not marked.


Comparison of antimicrobial resistant Escherichia coli in wild and captive Japanese serows. The fecal Escherichia coli isolated from wild Japanese serows living in mountainous areas away from humans and those from captive serows kept in human areas were examined for antimicrobial resistance and the possession of transferable R plasmids. Of 874 E. coli strains isolated from 283 wild serows in 1980-1981, only 11 (1.3%) were resistant to at least one of 6 antimicrobial drugs; ampicillin, streptomycin, tetracycline, chloramphenicol, kanamycin and sulfadimethoxin. Seven (2.5%) individuals were found to carry resistant E. coli. To heighten the isolation frequency of drug-resistant strains, fecal samples of 244 wild serows in 1983-1984 were cultured directly onto drug-supplemented media. Only 12 (4.9%) serows were shown to have drug-resistant E. coli. No transferable R plasmid was detected among a total of 87 resistant strains from wild serows. In contrast, all 33 captive serows except one which was kept only one day after capture, showed resistant E. coli and 20 (60.6%) serows were excreting R plasmid-carrying E. coli. Of 161 drug-resistant strains from captive serows, 50 (31.1%) were found to carry R plasmids. Wild serows seemed to readily change to harbor resistant E. coli almost as soon they were reared in human areas without direct exposure to drugs. These results lead to the conclusion that drug-resistant E. coli can probably be used as microbial indicator for natural environmental pollution.


Anatomically corrected malposition (ACM) with subpulmonary infundibulum in a calf: clinico-morphologic case report. A Japanese black calf with cyanosis, tachycardia, tachypnea and systolic murmur died of hypoxemia and cardiac insufficiency on the 38th day after birth. We could not establish the diagnosis during it's life. However, anatomically corrected malposition (ACM) with ventricular septal defect was confirmed at autopsy. There was situs solitus of the viscera and atria with atrio-ventricular discordance and ventriculo-arterial concordance. The ventricles demonstrated l-loop, i.e. on the right-sided ventricle there was a markedly enlarged morphologic left ventricle, and on the left-sided ventricle there was a hypoplastic morphologic right ventricle with a stenotic tricuspid valve and Ebstein-like deformity. The right posterior aorta originated from the left ventricle. The pulmonary artery arose from the left-sided right ventricle via infundibulum. There was a fibrous continuity between the aortic and mitral valve. We considered that this is the first reported case of bovine ACM.


The role of 36 megadalton plasmid of Salmonella enteritidis for the pathogenesis in mice. The pathogenesis associated with the 36 Megadalton (Md) plasmid of Salmonella choleraesuis subsp. choleraesuis serovar Enteritidis (S. Enteritidis) was assessed by using the plasmid-containing strain AL1190, plasmid-cured strain AL1192, and plasmid-reintroduced strain AL1193. After oral inoculation of strain AL1190 or AL1192 to C57BL/6 mice, mesenteric lymph nodes, spleens, and livers were examined for the numbers of viable bacteria and for the histopathological changes. The numbers of bacteria were greater and histopathological changes were severer in these organs of mice inoculated with strain AL1190, than in those with strain AL1192. Strains AL1190, AL1192, and AL1193 showed the equivalent survival rate to sera of guinea pig, calf, and pig, and grew equally well under the iron-limiting conditions. These results suggested that the 36 Md plasmid of S. Enteritidis contribute to the spread of the infection beyond small intestines to mesenteric lymph nodes, spleens, and livers, but not to serum resistance or acquisition of iron.


Chemotactic activity of hemocytes derived from a brackish-water clam, Corbicula japonica, to Vibrio parahaemolyticus and Escherichia coli strains. Hemocytes from adult and juvenile specimens of a brackish-water clam, Corbicula japonica, were attracted chemotactically to live cells of Vibrio parahaemolyticus and Escherichia coli strains in a balanced salt solution, which was enhanced significantly in the presence of respective C. japonica plasma. Chemotactic attractions of adult's and juvenile's hemocytes were seen also in artificial seawater at a similar level to those in the balanced salt solution. Chemotactic attractions of juvenile's hemocytes to these strains were lower in level than those of adult's hemocytes. C. japonica plasma seems to facilitate for C. japonica hemocytes to recognize these organisms.


Immunohistochemical findings of juxtaglomerular cells on acute phase in two-kidney Goldblatt hypertensive rats and spontaneously hypertensive rats. Juxtaglomerular (JG) cells on the acute phase in two-kidney Goldblatt hypertensive (2KGH) rats and spontaneously hypertensive rats (SHR) were examined immunohistochemically. JG cells in 2KGH rats and SHR were positively stained with anti-renin serum and anti-angiotensin II (A II) serum. In 2KGH rats, the number of renin and AII immunoreactive JG cells in the clipped kidneys increased throughout the observation period. The number of renin and AII immunoreactive JG cells in the unclipped kidneys was almost the same as that in control rats, although immunoreactivity of these cells was weak and they were small in size. These changes in the unclipped kidneys became obvious with the time course after operation. We did not see any changes in these cells in SHR. In 2KGH rats treated with captopril, the number of renin immunoreactive JG cells in the clipped kidneys increased, whereas that of AII immunoreactive JG cells in the bilateral kidney decreased. When captopril was administered to SHR, the number of renin immunoreactive JG cells in the bilateral kidney increased, whereas that of AII immunoreactive JG cells in the bilateral kidney decreased. These results suggested that the JG cell in the bilateral kidney was closely related to the development of hypertension in 2KGH rats, but not in SHR. The increase of renin immunoreactive JG cells in 2KGH rats and SHR treated with captopril was probably due to the removal of negative feedback inhibition of AII on JG cells.


Monoclonal antibodies to feline lymphocyte membranes recognize the leukocyte-common antigen (CD45R). By immunization of BALB/c mice with a feline T lymphoblastoid cell line, MYA-1 cells, two types of lymphocyte-specific monoclonal antibodies (mAbs) were obtained. The 220/205/190 kd protein defined by 2F11 mAb is highly expressed on the surface of MYA-1 cells and another feline T lymphoma cell line, FL74 cells. The protein is also expressed on normal feline thymocytes, splenocytes and feline peripheral blood mononuclear cells (PBMCs). Another mAb, 17B10, caused similar results as those of 2F11 except for its low reactivity with FL74 cells. The second type of mAb, 15B3, defined the 220 kd protein. The reactivities of this mAb with MYA-1 cells, FL74 cells, PBMCs and feline splenocytes were lower than the former two mAbs, and did not react to feline thymocytes. On the other hand, 17B10 and 15B3 defined partial populations of MYA-1 and FL74 cells recognized by 2F11. The cells defined by the 2F11 and 17B10 are all leukocytes in spleen and lymph node. In contrast, 15B3 defined most of the cells in B cell area and partially in T cell area. These results suggested that 2F11 and 17B10 recognized the specific antigen of 220/205/190 kd of the leukocyte-common antigen (L-CA) family, CD45R, with different epitopes, and that 15B3 defined the distinct antigen of 220 kd on CD45R.


Clinicopathology of meningoventriculitis due to Streptococcus bovis infection in neonatal calves. Three neonatal calves ranging in age from 4 to 14 days were examined pathologically and bacteriologically. The calves showed depression, anorexia, pyrexia, and difficulty or inability to stand followed by cloudiness of the ocular aqueous humor or cornea. Autopsy revealed congestion, petechiae, and cloudy areas in the meninges. Histologically, the central nervous system (CNS) lesions were prominent and limited to the meninges where fibrinous exudate and infiltrations of neutrophils, macrophages, and lymphocytes were present. There were mild or slight degrees of choroid plexitis and ependymitis. Endophthalmitis was seen as a concurrent lesion in all cases. Fibrinous or fibrinopurulent changes were found in the peritoneum and epicardium as well as in several other organs. Numerous Gram-positive cocci were detected in affected areas of the whole body. Bacteriologically, Streptococcus bovis was isolated from all examined materials consisting of the brain, cerebrospinal fluid, ocular aqueous humor, and several other organs. These results suggest that the lesions were associated with infection of the organism and that the present cases were in the process of septicemia.


Non-specific natural cytotoxic factor released from bovine peripheral blood lymphocytes. Natural cytotoxicity against bovine leukemia cells (PC-3 cells) was found in bovine peripheral blood lymphocytes (PBL), and in non-adherent cells but not in adherent cells to nylon-wool column. Natural cytotoxic cells (NCC), which have natural cytotoxic activity, are found in T cell-rich fraction. When NCC were cocultured with PC-3 cells, natural cytotoxic factor (NCF) was released rapidly from NCC, and dose-response curve for NCF was almost linear induction. Cytotoxicity against PC-3 cells by NCC or NCF was increased with an increment of incubation period. Cytotoxicity against K562 cells, CL-1 cells, M1 cells or EL-4 cells by NCF was almost the same level as that against PC-3 cells, but that against those cell lines by NCC was not found. NCF activity in culture fluid from NCC cocultured with K562 cells or CL-1 cells was lower than that from NCC cocultured with PC-3 cells.


Immunocytochemical and morphometric studies on the effects of dehydration on vasopressin-secreting cells in the hypothalamus of the Mongolian gerbils. Effects of dehydration on vasopressin-secreting cells (VP cells) of the supraoptic nucleus (SON) and the paraventricular nucleus (PVN) in the Mongolian gerbils were studied immunocytochemically and morphometrically. The plasma osmolality was measured at the time of sacrifice of individual animals and the body weight was measured every day during dehydration. The plasma osmolality increased significantly on day 3 of dehydration, followed by a gradual increase to reach nearly its equibilium state on day 10. The body weight decreased rapidly until day 10, followed by a gradual decrease thereafter. The area of VP cells increased significantly in both the SON and PVN on day 1 of dehydration, the level being nearly the same until days 3 to 5 and going up on day 7 to reach the plateau after day 15. These findings seem to reflect a compensation mechanism between the volume of body fluid and the plasma osmolality and to reflect responses of VP cells to the osmotic stimuli. Electron microscopic observation revealed that, at the beginning and late stages of dehydration, the increase in the area of VP cells was in parallel with the expansion of the Golgi area and with the distension of cisternae of the rough endoplasmic reticulum.


Experimental Tonsillophilus suis granuloma in rats and guinea pigs. Rats and guinea pigs were experimentally infected by intradermal and intraperitoneal inoculation of T. suis. These animals were observed for pathologic changes at various stages. Macroscopic observation disclosed visible grayish white soft nodules at the injection site in those inoculated intradermally and many grayish white nodules in the peritoneum of those inoculated intraperitoneally. Histopathological examination revealed the presence of characteristic microbial elements in the centre of the lesions. Various stages of the lesions were recognized; the early stage being abscess, the advanced stage being pus-forming granuloma and the final stage being residual granuloma. The microbial elements were composed of spores, thalli and multilocular tubers. Clubs were formed around microbial elements in rats inoculated intradermally and in guinea pigs inoculated intraperitoneally. The characteristic T. suis lesions in pigs were reproduced by experimental infection of this organisms in rats and guinea pigs, and thus the pathogenicity of T. suis was confirmed.


Relationship between pulmonary arterial pressure and pulmonary thromboembolism associated with dead worms in canine heartworm disease. To examine effects of thromboemboli due to dead worms on pulmonary arterial pressure (PAP), 20 to 50 dead heartworms were inserted into the pulmonary arteries of 4 heartworm uninfected dogs (uninfected group) and 11 dogs infected with heartworms (infected group). In the uninfected group, the mean PAP rose 1 week after worm insertion (10.9 to 166. mmHg), but it recovered by the 4th week. Clinical signs, hemodynamics and blood gas findings also deteriorated at the 1st week, but recovered at the 4th week. Angiographic and pathological findings indicated that blood flow recovered through the spaces between thromboemboli and vessel walls at the 4th week. The infected dogs were divided into three groups. In the infected-I group (5 dogs), the intimal lesions of the pulmonary arteries were slight, and clinical and laboratory findings showed changes similar to those of the uninfected group. In the infected-II group (4 dogs), the pulmonary arterial lesions were severe and the mean PAP was higher (25.7 mmHg) than in the uninfected group before worm insertion. An increase in PAP (34.1 mmHg) and worsening of clinical and laboratory findings were noticed till the 4th week. Thromboemboli adhered extensively to the vessel walls. Two dogs in the infected-III group died of severe dyspnea on the 9th and 10th day, and the mean PAP rose remarkably at the 1st week (from 19.4 to 28.2 mmHg). Severe pulmonary parenchymal lesions with edema or perforation were observed. From the above results, it was clarified that effects of dead worms on PAP and clinical signs depended on the severity of pulmonary arterial lesions before worm insertion.(ABSTRACT TRUNCATED AT 250 WORDS)


The complete amino acid sequence of the Clostridium botulinum type D neurotoxin, deduced by nucleotide sequence analysis of the encoding phage d-16 phi genome. The complete nucleotide sequence of Clostridium botulinum type D strain CB16 neurotoxin was determined and the deduced amino acid sequence is reported here for the first time. The structure and function of botulinum type D neurotoxin is discussed from a molecular biological viewpoint. DNA was extracted from toxin-converting phage d-16 phi of C. botulinum type D strain CB16, and a fragment (about 10 kbp) coding for the neurotoxin was cloned into Escherichia coli using lambda gt11. A 21-mer oligonucleotide which corresponds to Phe7 to Val13 of the partial amino acid sequence near the N-terminus of the type D neurotoxin was synthesized and used as a probe to identify the gene encoding type D neurotoxin. The nucleotide sequence contained a single open reading frame coding for 1,275 amino acids (molecular weight of 146,785) and the deduced amino acid sequence corresponded exactly to the partial amino acid sequences determined by direct microsequencing of the neurotoxin fragments. In the dichain molecule of the neurotoxin, Thr2 and Asn443 formed the N-termini of the light chain (M.W. 50,410) and heavy chain (M.W. 96,394) respectively, and these two chains were linked with a disulfide bond between Cys437 on the light chain and Cys450 on the heavy chain. The nucleotide sequence of the D-CB16 neurotoxin differed from that previously reported for type D neurotoxin by three nucleotides.


Overshoot and high equilibrium in body temperature responses of rats to ambient heat: relation to thermoregulatory ability and improvement in estimation of survival time in a hot environment. Physiological significance of body temperature responses to ambient heat (BTRAH) with high overshoot or a high equilibrium phase was studied in relation to thermoregulatory ability. Subsequently, an attempt was made to predict survival time (ST) by measuring the period required to attain a colonic temperature (Tco) of 42.0 degrees C (t42.0), whereas ST hitherto having been calculated on the basis of a period until Tco attains 42.5 degrees C (t42.5). Tco of rat was monitored continuously with a Cu-Co thermocouple during exposure to an ambient temperature (Ta) of 42.5 degrees C. The lower the overshoot temperature (Tos) or the equilibrium temperature (Teq), the longer the rats survived in a hot environment. The present findings further suggest that these two types of BTRAH are immature variations of the typical triphasic BTRAH which is characteristic of heat resistant individuals. A new regression line of ST (Y) as a function of the t42.0 (X) was obtained for most rats as follows: Y = 0.963X + 43.85 (male); and Y = 0.973X + 39.10 (female). This equation enabled to calculate ST without thermal death. However, the former approach based on t42.5 must be applied yet in small number of rats which showed the irregular BTRAHs with Tos or Teq higher than 42.0 degrees C. Delayed influence on survival and fecundity at a Ta of 24 degrees C were not found during one year following this hyperthermia up to 42.5 degrees C.


Theoretical epidemiology on bovine ephemeral fever outbreaks in Tanegashima Island, Kagoshima Prefecture of Japan in 1988. From the end of September to November 1988, a compact scale of bovine ephemeral fever (BEF) outbreaks occurred suddenly in Tanegashima island of Kagoshima Prefecture, southern part of Kyusyu island of Japan. The BEF outbreak pattern showed epidemical characteristics as follows; (1) outbreak spread from few foci to zone during one month, and (2) the disease might be transmitted in farms with a fixed probability of adequate contact. By using the above aspects, we attempted to analyze the disease theoretically with the application of Poisson distribution and Reed-Frost model. The BEF incidence in farms was in well accord with the Poisson distribution. As the very rare event occurred in unit time or in unit area in this epidemic, the cattle population at risk were equivalently susceptible to BEF virus in this island, due to the influence of no vaccination to BEF control before the first outbreak. Similarly, the epidemic curve of the Reed-Frost model was proved to fit well the incidence observed in a farm, and the probability of adequate contact was induced as p = 0.226. If the cattle population is less than 5 in this farm, the outbreak would not occur in the first instance.


Detection and characterization of Theileria sergenti proteinases. The lysate of Theileria sergenti piroplasms was tested for proteinases using sodium dodecyl sulfate-polyacrylamide gel electrophoresis in which substrate was included in gel matrix. Six proteinases of molecular weight 330, 125, 98, 94, 67 and 58 kilodalton (kDa) were detected. From the results of the Triton X-114 phase partition, 330, 125 and 58 kDa proteinases were partitioned into aqueous phase, which indicated that they were not associated with parasite membranes. All these three enzymes were classified into metalloproteinase family because of their sensitivities to metal-ion chelating compounds, ethylenediaminetetraacetic acid (EDTA) and 1,10-phenanthroline. On the other hand, 98 and 94 kDa proteinases were membrane-associated metalloproteinases which were preferentially inhibited by 1,10-phenanthroline. Another metalloproteinase of 67 kDa which was inhibited by EDTA and 1,10-phenanthroline was not associated with parasite membranes. Proteinases of 98 and 94 kDa degraded heat-denatured hemoglobin.


Pregnancy and lactation affect the microvasculature of the mammary gland in mice. Microvascular changes in the mammary gland in mice during pregnancy and lactation were investigated by scanning electron microscopy (SEM) with a corrosion cast method, transmission electron microscopy (TEM) and morphometry. By SEM, duct-associated capillary plexuses were sparsely distributed to branch into adipocytes during virgin period. With advance in pregnancy, both branches from the capillary plexuses and branches from the vessels surrounding adipocytes extended further to form capillary networks. The basket-like architecture was completed by day 18 of pregnancy. These findings may indicate that angiogenesis occurs frequently during this period. During lactation, the basket-like architecture still remained and the capillaries surrounding alveoli meandered. After weaning, the regression of microvasculature followed the degeneration of alveoli. By TEM and morphometry, the density of pinocytotic vesicles (PVs) (number of PVs per microns 2 of endothelium cytoplasm) increased twofold from day 18 of pregnancy to day 5 of lactation, furthermore increased threefold from days 10 to 20 of lactation, and subsequently decreased after weaning. Marginal folds and microvillous processes gradually increased in length with advance in pregnancy, reached the maximum from days 5 to 15 of lactation, and thereafter decreased. In addition, the capillaries with thinner walls were in close contact with alveoli during the late stage of pregnancy and during lactation. Furthermore, the alveolar epithelial cells had well-developed basal infoldings during lactation. These findings suggest that the capillaries play an important role in transporting materials necessary for milk production.


Changes in blastogenic responses of lymphocytes and delayed type hypersensitivity responses after vaccination in dogs. To clarify the immunologic effects of vaccination in dogs, we monitored total leukocyte and lymphocyte counts, humoral antibody responses, blastogenic responses of lymphocyte, and delayed type hypersensitivity (DTH) responses after vaccination. Mixed vaccines were administered on day 0 except for canine parvovirus (CPV) vaccine which was readministered on day 21. The puppy and adult dogs had a significant decrease in leukocyte and lymphocyte counts on day 7. The puppies showed a significant increase in the blastogenesis of lymphocytes after each vaccination, whereas the adult dogs had no significant changes. However, the adult dogs were divided into two groups, high responders and low responders in blastogenesis of lymphocytes. The dogs with higher or lower response in SI values on day 0 tended to show decrease or increase after the first vaccination, respectively. Since almost all dogs developed high titers of humoral antibody, it is considered that vaccination acts in an immunomodulative fashion. DTH responses to phytohemagglutinin (PHA) and CPV vaccine monitored at 0, 3, and 8 weeks after the first vaccination produced strong reactions, in particular those to CPV vaccine rose significantly after vaccination and maintained the higher responses for at least 2 months. These results suggest that DTH responses to PHA and CPV vaccine are helpful to monitoring non-specific and specific immune functions in vivo, therefore, DTH could be used as simple and rapid immunologic tests in canine practice.


Detection of uremic peaks in dogs by anion exchange high performance liquid chromatography. Sera of dogs with gentamicin-induced uremia were analyzed by high performance liquid chromatography system with strongly basic macroreticular anion exchange resin. Satisfactory separation of peaks was achieved with good reproducibility after deproteinization of sera with trichloroacetic acid at a final concentration of 3%, confirming that the system was suitable for qualitative analysis of uremic serum. The chromatograms showed that the number of peaks and the peak area had relation to concentrations of serum urea nitrogen or creatinine and severity of uremia. Four peaks were selected as suspected canine uremic peaks with high correlation to serum creatinine concentrations which were hardly influenced by extrarenal factors. The results suggested that these four fractions might contain uremic toxins.


Field trials on a live bovine respiratory syncytial virus vaccine in calves. Field trials were carried out in calves using a live bovine respiratory syncytial (BRS) virus vaccine prepared from the attenuated BRS virus, strain rs-52. Two hundred seventy-five and 353 calves were vaccinated intranasally and intramuscularly, respectively. No undesirable postvaccinal reactions were observed in the vaccinated calves. Of the serum neutralizing (SN) antibody negative calves 89.7% (26/29) and 92.8% (90/97) developed SN antibody 1 month after intranasal and intramuscular vaccination, respectively. Most of the calves having SN antibody titers of 1:1 or 1:2 at the time of vaccination showed a significant increase in SN antibody titer. About 70% and 90% of the calves vaccinated intranasally and intramuscularly, respectively, maintained SN antibody for 6 months after vaccination. In a field trial, a natural BRS virus infection occurred about 5 months after the start of the trial. Ten of the 16 unvaccinated control calves showed respiratory symptoms due to BRS virus infection. On the contrary, all of the 68 vaccinated calves exhibited no symptoms at all, indicating efficacy of the vaccine.


Glomerulopathy with IgA deposition in the dog. In 100 dogs autopsied, glomerular IgA deposition was examined by the immunofluorescence technique and the histopathological features of glomeruli with IgA deposition were examined by light and electron microscopy. The incidence of the IgA deposition was age-related but there were no sex and breed predisposition. Deposition of IgA was observed mainly in mesangial areas in approximately a half (47%) of dogs examined. IgG, IgM and C3 often co-deposited. Histopathology of the glomeruli with IgA deposition indicated increase of mesangial cells, crescent formation, hemispherical deposits in paramesangial areas and glomerular sclerosis. Ultrastructurally electron dense substances positive for IgA deposited in mesangial and paramesangial areas. The examination to know the relation between the severity of IgA deposition and the number of mesangial cells or percent of the cells to total glomerular cells indicated that mesangial cells increased at the early stage of the disease and subsequently epithelial and endothelial cells proliferated as the increasing amount of IgA. Dogs suffering from enteritis or liver diseases showed high incidence of glomerular IgA deposition.