Comparison of peripartum metabolic status and postpartum health of Holstein and Montbéliarde-sired crossbred dairy cows.

Objectives of the current experiment were to evaluate plasma concentrations of metabolites and haptoglobin peripartum, uterine health and involution, and follicle growth and resumption of cyclicity of Holstein (HO) and Montbéliarde-sired crossbred cows. Cows (52 HO and 52 crossbred) were enrolled in the study 45 d before expected calving date. Cows had body weight and body condition score recorded on d -45, -14, 0, 1, 28, and 56 relative to calving. Dry matter intake was calculated for a subgroup of cows (25 HO and 38 crossbred) from 6 wk before to 6 wk after calving. Blood was sampled weekly from d -14 to 56 relative to calving for determination of glucose, nonesterified fatty acid, and ß-hydroxybutyrate concentrations; from d -7 to 21 relative to calving for determination of haptoglobin concentration; and from d 14 to 56 postpartum for determination of progesterone concentration. Cows were examined at calving and on d 4, 7, 10, and 14 postpartum for diagnosis of postparturient diseases, on d 24 postpartum for diagnosis of purulent vaginal discharge, and on d 42 postpartum for diagnosis of subclinical endometritis. Uteri and ovaries were examined by ultrasonography every 3 d from d 14 to 41 postpartum. Milk yield and composition were measured monthly and yield of milk, fat, protein, and energy-corrected milk were recorded for the first 90 d postpartum. Body weight was not different between Holstein and crossbred cows, but HO cows had reduced body condition score compared with crossbred cows. Even though DMI from 6 wk before to 6 wk after calving tended to be greater for HO cows (16.8 ± 0.7 vs. 15.3 ± 0.5 kg/d), HO cows tended to have more pronounced decline in dry matter intake, expressed in percentage of body weight from d -15 to 0 relative to calving. Energy-corrected milk and nonesterified fatty acid and ß-hydroxybutyrate concentrations were not different between breeds. No differences were observed in incidence of retained fetal membranes, metritis, and subclinical endometritis, but HO cows tended to be more likely to have pyrexia from d 0 to 15 postpartum (50.0 vs. 31.4%) and to have greater incidence of purulent vaginal discharge (44.2 vs. 26.5%) than crossbred cows. Holstein cows were more likely to have at least 1 uterine disorder postpartum than crossbred cows (63.5 vs. 36.7%). No differences between breeds were observed in uterine involution. Holstein cows had larger subordinate follicles (10.1 ± 0.4 vs. 8.9 ± 0.5) and a greater number of class III follicles (1.6 ± 0.1 vs. 1.2 ± 0.1) than crossbred cows. Furthermore, the first corpus luteum postpartum of HO cows was diagnosed at a slower rate compared with crossbred cows. Crossbred cows had improved uterine health compared with HO cows and this may have been a consequence of heterosis and (or) breed complementarity and less pronounced decrease in DMI during the last days of gestation.

Effects of prepartum controlled-energy wheat straw and grass hay diets supplemented with starch or sugar on periparturient dairy cow performance and lipid metabolism.

This study examined the effects of a forage source [wheat straw (WS) versus grass hay (GH)] prepartum and supplemental carbohydrate source [corn (dry feed; DF) versus molasses (liquid feed; LF)] on pre- and postpartum intake, digestibility, selective particle consumption, milk yield, and lipid metabolism. The objectives were to determine if forage or pre- and postpartum supplement alters periparturient intake, energy balance, and milk yield. Sixty (n=15) multiparous dairy cows were used in a randomized complete block design with a 2 × 2 factorial arrangement of treatments to compare WS versus GH diets supplemented with either DF or LF. Dietary treatments were (1) WS prepartum + DF pre- and postpartum (WSDF), 2) WS prepartum + LF pre- and postpartum (WSLF), (3) GH prepartum + DF pre- and postpartum (GHDF), and (4) GH prepartum + LF pre- and postpartum (GHLF). Treatments began at dry-off, × before expected calving. During the prepartum phase, cows maintained dry matter intake (DMI) at 2.0% of body weight and prepartum energy balance remained positive for all treatments until calving. Prepartum GH diets had a more positive energy balance compared with WS diets. On week -5, energy balance was more positive for GHDF than for WSDF or GHLF. Energy balance for WSLF, however, was lower on week -3 and -1 than GHDF. Liquid feed decreased dry matter digestibility and increased prepartum liver triglyceride, serum nonesterified fatty acids (NEFA), and tended to increase ß-hydroxybutyrate. After calving, LF decreased DMI and energy balance, but not yield of milk or 3.5% fat-corrected milk, resulting in greater feed efficiency compared with DF. Forage did not affect postpartum DMI, but milk yield tended to be higher for WS versus GH. The DMI expressed as percentage of body weight was not affected by supplement or prepartum forage type. Cows fed WS had lower serum NEFA, higher liver glycogen, and tended to have a lower triglyceride to glycogen ratio postpartum than GH. Serum NEFA peaked on d 14 for all treatments and then declined thereafter. In postpartum diets, more particles were retained on the top screen for LF (>19.0mm) of the Penn State Particle Separator, which also tended to have more particles in the second screen (particles 19.0-8.0mm). Supplement had minimal effect on postpartum selective particle consumption. In conclusion, feeding diets containing WS resulted in lower postpartum serum NEFA, higher liver glycogen, and a tendency for greater milk production and lower liver triglyceride to glycogen than those containing GH. Liquid feed reduced postpartum DMI but not yield of milk yield or 3.5% fat-corrected milk yield, resulting in an improvement in feed efficiency. Future research should continue to investigate the use of single dry cow diet feeding strategies as they affect pre- and postpartum animal responses.

Effects of feeding wheat straw or orchardgrass at ad libitum or restricted intake during the dry period on postpartum performance and lipid metabolism.

The objectives of this study were to investigate the effects of forage source [wheat straw (WS) or orchardgrass hay (OG)] and total amount of diet dry matter fed [ad libitum or restricted to 70% of predicted dry matter intake (DMI)] prepartum on postpartum performance. The study design was a 2×2 factorial design with 10 cows per treatment. Treatments were WS total mixed ration (TMR) ad libitum, OG TMR ad libitum, WS TMR restricted, and OG TMR restricted. The WS TMR (dry matter basis) contained 30% WS, 20.7% corn silage, 10.0% alfalfa hay, 18.2% ground corn, 16.8% soybean meal, and 4.3% molasses mineral mix (14.7% CP, 1.5 Mcal/kg of net energy for lactation, 37.0% neutral detergent fiber). The OG TMR contained 30% OG, 46.2% corn silage, 10.0% alfalfa hay, 9.5% soybean meal, and 4.3% molasses (14.2% CP, 1.5 Mcal/kg of net energy for lactation, 41.0% neutral detergent fiber). Cows received 1 lactation diet after calving (17.7% CP, 1.6 Mcal/kg of net energy for lactation, 27.3% neutral detergent fiber). Total diet DMI prepartum was higher for ad libitum than for restricted as designed, but forage source had no effect on DMI. Total tract apparent digestibilities of DM and NDF were greater for OG than for WS. Postpartum DMI expressed as a percentage of body weight for the first week of lactation was higher for ad libitum than for restricted diets. Postpartum DMI during the first 30 d of lactation was higher for OG than for WS, but no effect was observed for the amount fed prepartum. Milk yield during the first week of lactation was higher for OG than for WS; however, during the first 30 d, 3.5% fat-corrected milk yield and yield of milk fat were highest for OG TMR restricted and WS TMR ad libitum. Prepartum treatments had a limited effect on pre- and postpartum lipid metabolism; however, cows fed WS TMR ad libitum had the highest postpartum ß-hydroxybutyrate. Eating behavior was observed by 10-min video scans of 24-h video surveillance for 5d pre- and postpartum. Prepartum eating time and eating bouts tended to be greater by WS than for OG, and postpartum eating time per kilogram of neutral detergent fiber intake tended to be greater for WS than for OG. Results indicate that forage source and amount of DM fed prepartum affected postpartum performance and tended to alter the behavior of cows in tie-stall barns.

Yield response of lactating Holstein dairy cows to dietary fish meal and bone meal.

Experiments were conducted to examine the effects of the source and amount of dietary protein on yield and composition of milk from Holstein dairy cows. Study 1 used 36 multiparous cows at 125 +/- 59 d in milk in, a replicated 2 x 2 Latin square design. Treatments were diets formulated to contain 16% crude protein (CP) in which 11% was fish meal or meat and bone meal supplied 11% of dietary CP. Intakes of dry matter, CP, and net energy for lactation; yields of milk; and percentage of milk fat were not affected by treatment. Fish meal increased contents of milk total N, casein N, and noncasein N but did not increase contents of NPN; fish meal also tended to increase milk CP yields. Study 2 used 78 cows (31 primiparous) at 31 +/- 2 d in milk in a randomized block design. Two treatment diets were formulated to contain 16 or 18.5% CP, and soybean meal was the sole source of supplemental protein in those diets. The two other treatment diets were, formulated to contain 16% CP; in these diets, fish meal or meat and bone meal partially replaced soybean meal. Treatments did not influence yield or composition of milk from multiparous cows. Compared with a soybean meal diet containing 16% CP, a soybean meal diet containing 18.5% CP or diets containing 16% CP and containing meat and bone meal or fish meal increased the milk, yield of primiparous cows similarly. Fish meal or meat and bone meal increased the efficiency of protein utilization for milk yield.

Responses of dairy cows supplemented with somatotropin during weeks 5 through 43 of lactation.

Beginning at wk 5 of lactation, 136 cows (34 per treatment) were supplemented daily for 38 wk with 0, 10.3, 20.6, or 41.2 mg of recombinantly derived bST monomer. Cows were obtained from University of Kentucky, University of Minnesota, University of Pennsylvania, and The Ohio State University. Nine cows (4 at 0 mg/d, 1 at 10.3 mg/d, 1 at 20.6 mg/d, and 3 at 41.2 mg/d) did not complete the experiment because of health problems. Data from these cows were included in the reproduction and health databases but not in the production database. Cows supplemented with bST produced more milk, consumed more feed, had lower rates of BW gain, and had improved efficiencies of milk production (conversion of feed and NEL to milk). Additional increases in productivity were modest at 20.6 and 41.2 mg/d versus productivity at 10.3 mg/d of bST. Concentrations of fat, protein, and TS in milk were unaffected. At 10.3 mg/d, bST did not adversely affect reproduction or health.

Early lactation responses of Holstein cows fed a rumen-inert fat prepartum, postpartum, or both.

Primiparous (n = 22) and multiparous (n = 41) cows were randomly assigned by calving date and parity to one of four dietary sequences of supplemental fat from 14 d prepartum to 151 d postpartum. Partially hydrogenated tallow was added to diets at 0% prepartum and postpartum (control); 1% prepartum and 2% postpartum; 0% prepartum and 2% postpartum; and 0% prepartum, 0% from 1 to 34 d postpartum, and 2% from 35 to 151 d postpartum. Inclusion of partially hydrogenated tallow did not influence yields of milk or 3.5% FCM, milk composition, or DMI during the first 151 d postpartum. During the first 35 d postpartum, cows receiving partially hydrogenated tallow starting at parturition yielded milk with a higher fat content than those receiving fat prepartum and postpartum. Addition of partially hydrogenated tallow to diets starting 35 d postpartum resulted in cows being more persistent in yields of milk and 3.5% FCM from 60 to 151 d postpartum. Reproduction parameters measured were unaffected by time of fat addition to diets. Our data suggest that delaying the addition of partially hydrogenated tallow to diets until 35 d postpartum may improve the persistency of lactation.

Effects of calcium salts of long-chain fatty acids added to a diet containing choice white grease on lactation performance.

We studied the effect of increased energy density of diets during early lactation on lactational performance of cows injected with bST. Total mixed rations contained 50% forage (DM) and 2.5% choice white grease during the first 14 wk of lactation. Thirty cows were fed diets with grease as the only supplemental fat, and 15 cows were fed grease plus 1.8% calcium salts of long-chain fatty acids. One-half of the cows fed grease and all cows fed grease plus calcium salts of long-chain fatty acids received 640 mg/28 d of sustained-release bST from 6 to 44 wk postpartum. Calcium salts of long-chain fatty acids did not increase milk yield, 3.5% FCM, or DMI during the first 14 wk postpartum or during 15 to 44 wk postpartum. Administration of bST increased milk fat concentration .4% units during the first 14 wk postpartum. Cows receiving bST had higher DMI and produced 18.5% more 3.5% FCM from 15 to 44 wk postpartum than did controls. Calcium salts of long-chain fatty acids or bST injections did not influence body condition score or BW. These data suggest that cows yielding 30 to 35 kg of milk/d during the first 14 wk of lactation may not respond to added fat over 2.5%.

Evaluation of calf starters containing different amounts of crude protein for growth of Holstein calves.

Holstein calves (n = 110) were used to evaluate the effect of calf starters containing 15, 16.8, 19.6, or 22.4% CP, DM basis (diets A, B, C, and D, respectively), on calf performance from d 4 to 56 of life. Preweaning daily gain tended to increase linearly as protein content of diets increased, averaging .37, .39, .38, and .44 kg/d for diets A, B, C, and D, respectively. After weaning, calves fed diet C gained the most (.86 versus .71, .75, and .79 kg/d for A, B, and D, respectively). Overall BW gains from d 4 to 56 averaged .54, .56, .62, and .61 kg/d for A, B, C, and D, respectively. Throughout the experiment, starter consumption tended to increase as CP content of diet increased. Under these conditions, maximum growth was supported by diet C (19.6% CP); no advantage was gained from higher (22.4%) protein content. Calf growth was moderate when calf starters of lower protein contents (15 or 16.8%) were fed.

Multi-farm use of bovine somatotropin for two consecutive lactations and its effects on lactational performance, health, and reproduction.

In a two-lactation study, 352 Holstein cows (124 primiparous) from six commercial dairy herds were assigned to daily injections of bST. Dosages were 0 (control), 5.15, 10.3, or 16.5 mg/d of bST; injections began 28 to 35 d postpartum. During yr 1, FCM production increased linearly as bST dose increased for primiparous and multiparous cows. However, FCM production for primiparous cows that were retained for yr 2 was not increased by bST, and the increase in FCM for multiparous cows was only 67% of that observed during yr 1 for doses of 10.3 and 16.5 mg/d. Milk SCC were not increased with the use of bST during either lactation. Days to conception of multiparous cows during yr 1 tended to increase linearly with increasing dose. Multiparous cows administered bST during yr 2 tended to have a lower conception rate than control cows (71 vs. 87%, respectively). Body condition was decreased linearly by bST dose at the end of yr 1 and 2. Prior to receiving bST during yr 2, bST-treated cows regained less body condition than control cows. Number of health disorders at parturition and assisted births of cows that received bST during yr 1 were unaffected by previous treatment. General health, ratio of single to multiple births, and BW gain of calves was similar among treatments.

Lupins (Lupinus albus) as a protein supplement for lactating Holstein dairy cows.

Three experiments were conducted to determine optimal substitution of lupin for soybean meal in dairy cattle diets; the effects of whole versus ground lupins and the effects of whole lupins versus whole soybeans on milk production were examined. In the first experiment, 57 cows were assigned randomly to treatments in blocks of five by calving order within parity. Treatments began 22 d postpartum and continued through d 140. Diets were isonitrogenously balanced using soybean meal as control: lupins replaced 0, 25, 50, 75, and 100% of supplemental protein. Cows that consumed 75% of supplemental protein as lupins produced more 3.5% FCM and fat than cows that consumed the control diet. In a second experiment, 30 primiparous cows were assigned randomly by calving order to either ground or whole lupins (60% of supplemental protein). Treatments began 32 +/- 3 d after calving and lasted for 84 d. Cows that consumed ground lupins produced more milk and 3.5% FCM than cows that consumed whole lupins. In the third experiment of 84 d, 28 primiparous midlactation cows were assigned randomly to diets containing whole lupins or whole soybeans to supply 1 kg of supplemental protein. Cows fed whole lupins produced milk with higher fat content, more 3.5% FCM, and more fat than cows consuming whole soybeans. Lupins are an acceptable supplemental protein source for dairy cows.

Influence of forage type, ratio of forage to concentrate, and methionine hydroxy analog on performance of dairy cows.

In a 2-yr study, 44 primiparous and 77 multiparous cows were assigned to one of 12 dietary treatments (2 X 3 X 2 factorial). Treatments were two forages (alfalfa or smooth bromegrass), three percentages of grain (40, 50 or 60% of diet DM), and two percentages of methionine hydroxy analog (0 or .15% of diet DM). Feeds were offered as total mixed diets. Data collection began 4 d postpartum and continued through 116 d postpartum. Dry matter intake was not affected by percentage of concentrate or forage source even though NDF of the diets ranged from 25.6 to 48.8% and ADF ranged from 15.7 to 36.8%. Cows fed bromegrass hay produced 1.5 kg/d more FCM and 1.2 kg/d more SCM than those fed alfalfa hay. Concentrate percentage in the diet increased milk yield (28.9, 30.4 and 31.3 kg/d at 40, 50 and 60%, respectively). Methionine hydroxy analog increased milk fat percentage and yield for cows fed diets of 50 and 60% concentrate with alfalfa hay but not for those fed diets of 50 and 60% concentrate with bromegrass hay. Effect of methionine hydroxy analog was not significant for milk fat or yield when diets of 40% concentrate were fed.

Somatotropin treatment for a second consecutive lactation.

Twenty-five Holstein cows were used to examine effects of recombinant bST administration for a second successive lactation on milk yield and composition, BW change, body condition, and health. Treatments were 0 (physiological saline), 10.3, 20.6, and 33 mg bST/d (8, 8, 7, and 3 cows per treatment, respectively). Saline and bST were given daily as subcutaneous injections beginning 4 to 5 wk postpartum and continuing for 38 wk. Somatotropin increased 3.5% FCM yield linearly from 8 to 36% over controls (control FCM was 30.1 kg/d). Milk composition was not affected by bST except that milk from cows given the 10.3-mg dosage had slightly higher Ca and P contents than did milk from cows receiving 0 or 20.6 mg. Cows receiving bST were 9 to 30% more efficient than controls in converting feed to milk. Body weight gain was unaffected by treatment. Average condition score was unchanged for control cows during the first 84 d following initiation of treatment, but declined in cows given bST. Thereafter, condition score increased in a similar manner for all groups. No treatment-related patterns were found in incidence of mastitis, feet and leg problems, ketosis, or milk fever. Blood hormone, chemistry, and hematology revealed no effects of bST treatment other than to raise plasma bST concentrations. Under conditions of this experiment, use of bST during a second consecutive lactation did not change the normal physiology or productive capacity of lactating cows.

Radiolabeled, nonspecific, polyclonal human immunoglobulin in the detection of focal inflammation by scintigraphy: comparison with gallium-67 citrate and technetium-99m-labeled albumin.

The accumulation of nonspecific polyclonal human immunoglobulin (IgG) radiolabeled with 125I or 111In was compared to that of [67Ga]citrate and [99mTc]albumin in rats with deep thigh inflammation due to Escherichia coli infection. Serial scintigrams were acquired at 1, 3, 24, and in some cases, 48 hr after injection. As early as 3 hr postinjection, [111In]IgG showed greater accumulation at the lesion than [99mTc]HSA (p less than 0.01). Both [125I]IgG and [111In]IgG showed greater accumulation than [67Ga]citrate (p less than 0.01). At 24 hr, IgG image definition increased, while HSA image definition decreased, and the intensity of accumulation of both IgG preparations was greater than that of [67Ga]citrate or [99mTc]HSA (p less than 0.01). At all imaging times, [67Ga]citrate accumulation was surprisingly low. In inflammation produced by Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, Candida albicans, or turpentine, [111In]IgG accumulation was similar to the results obtained with Escherichia coli. These studies suggest that focal sites of inflammation can be detected with radiolabeled nonspecific human polyclonal IgG.

Specific and nonspecific imaging of localized Fisher immunotype 1 Pseudomonas aeruginosa infection with radiolabeled monoclonal antibody.

To determine if radiolabeled specific antibodies directed against bacterial antigens could be used to detect sites of infection, gamma camera imaging studies were performed in animals infected with Pseudomonas aeruginosa. Murine monoclonal antibodies (Mabs) directed against Fisher Immunotype 1 Pseudomonas aeruginosa and a nonmicrobial, nonmammalian haptene, p-arsanilic acid, were labeled with 125I by the lodogen-Bead method. Unilateral, deep thigh infections were created by innoculation with 2 X 10(8) Fisher Immunotype 1 P. aeruginosa. Twenty-four hours later, one of the radiolabeled antibodies was injected intravenously at a dose of 0.25 mg/kg (100-150 microCi). Serial gamma imaging was then carried out beginning at 4 hr and at approximately 24-hr intervals thereafter. Beginning as early as 4 hr postinjection, the area of inflammation could be visualized with either the specific or nonspecific Mab, with the images continuing to intensify until 24-48 hr postinjection. At 48 hr, the contrast between lesion and background with the nonspecific Mab began to fade, while the contrast in the specific Mab-generated images continued to intensify until approximately 192 hr postinjection. Clear-cut differentiation between specific and nonspecific Mab-generated images was possible by 72 hr postinjection. We conclude that specific immune imaging of localized infection with Mab's directed against specific microbial antigens is possible and should be clinically useful. In addition, images created by the localization of immunoglobulin non-specifically at the site of inflammation in the first 24-48 hr postinjection may also provide useful information as to the anatomic location of hidden abscesses.

Effects of recombinant bovine somatotropin on milk production, body composition, and physiological parameters.

Twenty-eight multiparous Holstein cows were utilized to determine effects of long-term administration of recombinant bovine somatotropin on lactational performance, body condition, body composition, hormones, blood constituents, and physiological parameters. Treatments were 0 (control), 10.3, 20.6, and 41.2 mg recombinant bovine somatotropin administered daily as subcutaneous injections beginning 4 to 5 wk postpartum and continuing for 38 wk. A total mixed diet of 28% corn silage, 22% alfalfa hay cubes, and 50% corn-soybean meal-based grain mixture was fed. Fat-corrected milk yields were increased 12 to 25% for cows treated with somatotropin as compared with controls (29.9 kg/d). Milk composition was similar among treatments. Cows receiving somatotropin consumed 4 to 10% more feed and were 11 to 17% more efficient than controls for conversion of feed to milk. Body weight changes were not significantly different among treatments, but cows receiving somatotropin gained 4 to 10% less weight than controls. Body fat (kg) of cows receiving 20.6 and 41.2 mg/d was less than that of cows receiving 10.3 mg/d or no somatotropin. Estimated weights of body protein and mineral, most blood constituents, respiratory rates, and body temperature were not affected by somatotropin administration. Plasma fatty acids were elevated and hematocrit values were reduced. Plasma insulin, serum somatotropin, and heart rate increased concurrently with somatotropin administration.

Diagnosis of renal proximal tubular injury by urinary immunoassay for a proximal tubular antigen, the adenosine deaminase binding protein.

A sandwich ELISA assay has been formatted from two commercially available murine monoclonal antibodies, URO-4 and URO-4a, directed against a 120,000 dalton glycoprotein, the adenosine deaminase binding protein (ABP), found on the brush border of the renal proximal tubular epithelial cell. Untimed urine samples from 37 normal individuals and urinary ABP less than 0.1 AU; 37 patients with pure glomerular disease had ABP less than 0.4 AU (with 29, or 76% less than 0.2 AU); 10 patients with pre-renal azotaemia had ABP less than 0.6 (with 8, or 80% less than 0.3 AU). In contrast, 79 patients with post-ischaemic acute tubular necrosis had ABP greater than 0.6 AU. Acute renal failure due to myoglobinuria, contrast dye, and aminoglycoside toxicity were all associated with urinary ABP greater than 1.0 AU. In addition, all six patients with acute bacteraemic pyelonephritis had ABP greater than 0.7 AU, as opposed to ABP less than 0.2 AU in the urines of 12 women with acute cystitis. We conclude that this monoclonal antibody based urinary assay is a sensitive measure of renal proximal tubular injury, reliably distinguishes acute tubular from glomerular disease, and may be helpful in differentiating forms of urinary tract infection.

Diagnosis of tubular injury in renal transplant patients by a urinary assay for a proximal tubular antigen, the adenosine-deaminase-binding protein.

Two murine monoclonal antibodies (URO-4 and URO-4a)--which detect different epitopes of a proximal tubular cell glycoprotein antigen, the adenosine-deaminase-binding protein (ABP)--have been formatted into a sandwich enzyme immunoassay for detection of ABP in the urine. Serial urine samples from 34 renal transplant patients during the first six months posttransplant were analyzed to determine the correlation of this test with clinical rejection and cyclosporin (CsA) nephrotoxicity. In 29/29 acute rejection episodes the ABP level was elevated, beginning 1-7 days prior to treatment of rejection. Eighteen patients were treated for rejection with courses of OKT3 or antithymocyte globulin: 0/6 whose ABP level fell to normal during therapy had rerejection; 10/12 whose ABP level remained elevated had rerejection within 7 days of therapy completion. Of 15 patients treated with CsA, 7 had no rejection or drug toxicity; all 7 had normal ABP levels. The remaining 8 had CsA nephrotoxicity, all in association with elevated ABP levels that rapidly fell to normal with decreased CsA dose. An additional 7 patients with creatinine elevations more than 6 months posttransplant were studied: 5 had chronic vascular changes on biopsy, no response to increased immunosuppression, and normal ABP levels; 2 had a cellular infiltrate on biopsy, response to increased immunosuppression, and elevated ABP levels. We conclude that the urinary ABP assay provides information useful in the management of renal transplant patients with acute and chronic rejection and CsA toxicity.

Competitive idiotype--anti-idiotype immunoassay for adenosine deaminase binding protein in urine.

This competitive immunoassay, based on inhibition by antigen of the idiotype-anti-idiotype interaction, detects adenosine deaminase binding protein (ABP), and involves use of monoclonal anti-idiotype antibodies prepared to a monoclonal antibody specific for ABP. The conditions for this new type of competitive immunoassay are investigated. This competitive immunoassay is as sensitive and reproducible as an earlier described "sandwich"-type immunoassay for ABP (Clin Chem 31: 679-683, 1985). Evaluation of urine samples from normal subjects and from patients showed increased concentrations of ABP in patients with renal disease.

Changes in immunoregulatory lymphocyte populations in patients with histoplasmosis.

Circulating T-lymphocyte subpopulations were enumerated in 65 patients with histoplasmosis and correlated with the different clinical manifestations of the disease. Acute pulmonary histoplasmosis, rheumatologic, disseminated, and chronic inflammatory manifestations of histoplasmosis were all associated with a significant elevation above normal of OKT8+ (suppressor-cytotoxic) lymphocytes and a significantly lower than normal OKT4+ (helper-inducer)-lymphocyte to OKT8+-lymphocyte ratio. In contrast, cavitary disease was associated with an increase in OKT4+ lymphocytes, a decrease in OKT8+ lymphocytes, and a higher than normal OKT4/OKT8 ratio. Clinical recovery was associated with normalization of these values. Functional activity determined by coculture techniques correlated closely with T-lymphocyte subset measurements. These distinct subset abnormalities may help monitor immunological aspects of disease activity.

A new method for cell volume measurement based on volume exclusion of a fluorescent dye.

Several methods currently in use for measuring mean corpuscular volume include: centrifuged packed cell volume, electronic impedance, and light scattering methods. Although these techniques are widely used and accepted, there are problems inherent to each method which may produce systematic errors that are difficult to estimate. This paper describes a new flow cytometric method of cell volume determination, based on the principle of volume exclusion, which may overcome the systematic errors of the methods currently in use. This method requires that the cells be suspended in a fluorescent dye which is unable to penetrate the cell membrane. The level of fluorescence which is produced when a narrow stream of the cell suspension is excited by a focused laser beam will remain constant until a cell arrives in the illuminated region thereby causing a decrease in fluorescence which is directly proportional to the cell's volume. The volume exclusion method is shown to give an estimate of mean red cell volume which correlates well with existing methods.