Effect of rejuvenation and frozen storage on 42-day-old AS-1 RBCs.

BACKGROUND: The FDA has approved a 42-day storage period for RBCs stored in ADSOL (AS-1). This study was undertaken to provide data for the FDA about the feasibility of salvaging AS-1 RBCs at the end of their storage period by rejuvenation and freezing. STUDY DESIGN AND METHOD: The investigation, consisting of a study (n = 10) and control (n = 6) arm, was carried out in two centers. In both centers, eight healthy volunteers donated a unit (450 mL) of whole blood. The RBC concentrates were stored at 4 degrees C in AS-1 for 42 days. The study units were rejuvenated, whereas the control units were not. All units were stored frozen at -80 degrees C, then deglycerolized and kept for an additional 24 hours at 4 degrees C. RESULTS: After the 42-day storage period, ATP had declined to 62 percent of the original value, 2,3 DPG was zero, and MCV was significantly larger than that of fresh RBCS: Following rejuvenation and deglycerolization, the mean ATP level was 141 percent, the mean 2,3 DPG level was 109 percent, and the MCV was normal. The freeze-thaw-wash recovery of the rejuvenated and nonrejuvenated RBCs was similar, 88.4 and 84.0 percent, respectively. There was no difference in hypoxanthine, inosine, and uric acid levels in the rejuvenated and nonrejuvenated units, which indicated that the chemicals in the rejuvenation solution and their by-products had been removed during processing. In both centers, the mean 24-hour survival of rejuvenated, deglycerolized RBCs exceeded 75 percent, whereas that of nonrejuvenated RBCs did not. The long-term survival rates of viable study and control RBCs were similar. CONCLUSION: Forty-two-day-old AS-1 RBCs that have been rejuvenated and then frozen have more than 75 percent viability and normal oxygen delivery function. Rejuvenation of RBCs does not introduce additional safety hazards to blood transfusion.

An irreducible knee dislocation: a case report.

An 8-year-old man presented after sustaining an injury during a fall. A closed reduction attempt failed, and after several tests, an open reduction was performed. With posterolateral dislocation of the knee, there can be anterior cruciate ligament, posterior cruciate ligament, and medial collateral ligament disruption. At the 6-month (final) follow-up, the patient had no subjective pain or instability. With this type of injury, the approach can be conservative monitoring or repair of all of the ligaments. Because of the age and activity level of our patient, we opted for repair of the medial collateral ligament initially with the possibility of late anterior cruciate ligament and/or posterior cruciate ligament reconstruction.

Ultrasonography in the management of developmental dysplasia of the hip (DDH).

Real time ultrasonography, according to the method of Graf, was performed on 113 infant hips because of abnormal physical findings (ranging from hip clicks to frank dislocation). Three parameters were measured on the standard coronal images: alpha and beta angles of Graf and the d/D ratio of Morin. Results indicated that the presence of hip clicks (88 hips) in general is a benign condition and not associated with abnormal anatomy. Hips that were reduced at birth but dislocatable (Barlow hip) showed no significantly abnormal acetabular anatomy. Hips that were dislocated at rest but were reducible (Ortolani hip) showed definite abnormal acetabular anatomy and femoral head coverage. Ultrasonography is beneficial in the management of developmental dysplasia of the hip (DDH); it confirms the clinical findings and eliminates radiation exposure to the infant's pelvis, especially in the first 4-6 months of life when standard radiography is not always reliable in diagnosing DDH.

The turning of American psychiatry.

Philosophic changes and the "re-medicalization" of psychiatry in the past half century are associated with: evolution of the biopsychosocial model; new techniques for biologic studies; drug treatment for mental disorders; funding for research and training; and development of a criteria-based nosology for psychiatric disorders. The last development owes much to the work of four Washington University faculty members, beginning in the 1950's. The second part of this paper will examine their early careers and the institutional policies that fostered their work.

Growth and reproductive performance of ewe lambs fed corn or soybean meal while grazing pasture.

In a 2-yr study a total of 120 Suffolk and Suffolk-cross ewe lambs, approximately 3 mo of age, were assigned within weight strata to one of three treatments: 1) control, pasture only (C), 2) .09 kg soybean meal/(hd.d) (SBM) plus pasture or 3) .45 kg whole shelled corn/(hd.) plus pasture to compare growth and reproductive performance and to determine lamb selectivity of forage quality and type. Each year, supplements were offered from June through the 3rd wk in November. Ewe lambs were managed as one flock on Kentucky bluegrass (Poa pratensis L.) and tall fescue (Festuca arundinacea Schreb.) pastures except for a brief daily supplementation period when all lambs were brought into a corral and sorted into treatment groups for the feeding. Ewe lambs were exposed to fertile rams from October 1 through the 3rd wk of November each year. Growth rates of ewe lambs grazing pasture alone were slower than those of corn-supplemented ewe lambs and were comparable to those of SBM-supplemented ewe lambs, but reproductive performances were similar (P greater than .05). Date of first detected rise in serum concentrations of progesterone and date of first observed estrus and lambing performance were similar (P greater than .05) among treatments for each year. Compared to hand-clipped forage, forage samples collected by esophageally fistulated lambs were higher (P less than .05) in crude protein and lower (P less than .05) in acid detergent fiber each year, indicating that lambs selectively grazed higher-quality forage. These data indicate that neither protein nor energy limited growth or reproductive performance of these ewe lambs.

Isoproterenol inhibits the increase in microvascular membrane permeability produced by bradykinin.

Bradykinin (BRADY) is hypothesized to cause the "capillary leak" syndrome in patients with sepsis, trauma, and burns. Our purpose was to determine if isoproterenol (ISO) reversed a BRADY-produced accelerated loss of intravascular fluid and protein into the interstitium of skin. An increase in microvascular permeability in canine hind paw skin was sustained by a continuous femoral artery infusion of BRADY (0.2 micrograms/kg/min). After 2 hours of BRADY, skin lymph flow (LYM FLOW microliters/min) increased nine-fold and skin lymph-to-plasma total protein concentration ratio (RTP) was substantially increased. Mean blood flow in the femoral arteries was increased four-fold by the BRADY infusion. After 2 hours of BRADY-induced increased permeability, five of the ten dogs were started on intravenous ISO (2 micrograms/min continuously) which increased heart rate from 182 +/- 15 to 222 +/- 11 beats/min. ISO reversed the increase in RTP produced by the BRADY. After 8 hours of BRADY, there was less tissue albumin in the dogs given ISO (14.5 +/- 2.0 vs. 29.5 +/- 6.6 mg/gram dry wgt, p less than 0.05 unpaired t-test). ISO can reverse the sustained increase in skin microvascular permeability produced by BRADY.

Relationships among intramuscular collagen, serum hydroxyproline and serum testosterone in growing rams and wethers.

Concentration and maturation of collagen and serum concentrations of hydroxyproline and testosterone were determined in growing rams and wethers to characterize developmental changes in collagen associated with a representative testicular steroid. Groups of eight rams and eight wethers were slaughtered at 12, 18, 24 and 30 wk of age. Concentrations of collagen in longissimus, supraspinatus and infraspinatus muscles and serum hydroxyproline were greater (P less than .05) in rams than in wethers at all ages. Collagen stability, as measured by collagen solubility and thermal shrinkage temperature, was greater (P less than .05) in wethers than in rams. Differences in collagen stability and serum hydroxyproline concentration indicated that collagen synthesis and turnover were more rapid in rams than in wethers. Serum hydroxyproline decreased (P less than .05) and collagen solubility decreased (P less than .05) with age, indicating that collagen turnover was occurring most rapidly in 12-wk-old lambs and that collagen maturation was predominant in 24- to 30-wk old lambs. Testosterone parameters measured in rams were unrelated within groups to collagen characteristics, possibly reflecting the high variability in testosterone secretion and the slow development of collagen. However, rams as young as 12 wk of age were under the influence of testosterone, and differences in collagen between rams and wethers were apparent at that time.

Effect of melatonin on induction of estrous cycles in anestrous ewes.

To examine the influence of melatonin on seasonality of reproduction, 97 multiparous Suffolk and Suffolk-cross ewes were randomly assigned to one of four treatment (TRT) groups in a 2 X 2 factorial arrangement. Treatments were as follows: 1) control (C); 2) melatonin (M); 3) progestogen (P) implant of norgestomet plus pregnant mare serum gonadotropin (PMSG) injection (P + PMSG) and 4) M plus TRT 3 (M + P + PMSG). From April 3 to June 24 an oral dose of 2 mg M was administered once daily at 1600 to each ewe in TRT groups M and M + P + PMSG. On April 30, ewes in groups P + PMSG and M + P + PMSG were implanted in the ear with 2 mg norgestomet for 13 d. Immediately following implant removal, each ewe was injected with 500 IU PMSG. Blood samples were collected from all ewes twice weekly from March 22 through June 24. Number of estrous cycles per ewe during the TRT period of 82 d (April 3 to June 24) was higher (P less than .05) for M + P + PMSG (2.1 +/- .2) than for C (.3 +/- .2), M (1.5 +/- .2) and P + PMSG (1.1 +/- .2). Control ewes had fewer (P less than .05) estrous cycles per ewe than either M or P + PMSG. Following the induced estrus, 40% of ewes in the M TRT had one estrous cycle; 32% had two or more cycles. For ewes treated with M + P + PMSG, 24% had one cycle, and 32% had two estrous cycles.(ABSTRACT TRUNCATED AT 250 WORDS)

Increased skin lymph protein clearance after a 6-h arterial bradykinin infusion.

When bradykinin (0.15-0.28 micrograms.kg-1.min-1) was infused into both femoral arteries of 11 anesthetized dogs, skin lymph flows increased by 25-371% within 2 h, and mean lymph protein concentrations increased by one-third. To determine whether, in addition to the initial increase in permeability, a 6.5- to 10-h bradykinin infusion caused a sustained effect, the bradykinin infusion into one hindpaw was stopped after 2 h (2HR), whereas the contralateral hindpaw was infused continuously (CONT). Two hours after the bradykinin infusion was stopped, Ringer lactate equal to 10% of the dog's body weight was given intravenously to further increase lymph flow. After Ringer lactate infusion, increase in lymph protein clearance from the CONT hindpaws was greater than that from the 2HR hindpaws (change in clearance from before Ringer lactate infusion to final: 2HR, 6.9 +/- 1.4 to 8.8 +/- 1.1; CONT, 23.4 +/- 2.5 to 40.2 +/- 4.8 microliters/min). In the final lymph samples of the CONT, but not 2HR, hindpaws, the lymph-to-plasma ratio for immunoglobulin G and immunoglobulin M divided by the albumin lymph-to-plasma ratio exceeded the value of these ratios in the base-line samples. An intravenous bolus of Evans blue dye was given less than 2 h before the end of the experiment. The concentrations of dye in the final lymph samples were greater in CONT hindpaws (12.6 +/- 3.7% plasma equivalents) than in the 2HR hindpaws (1.1 +/- 0.5%). A continuous 6.5- to 10-h intra-arterial bradykinin infusion produced a sustained increase of transvascular protein clearance in skin that is consistent with a sustained increase in microvascular membrane permeability.

Reproductive performance of Suffolk and Suffolk-cross ewes and ewe lambs exposed to vasectomized rams before breeding.

Multiparous Suffolk and Suffolk-cross ewes were randomly allotted to treatments within breed and year to measure effects of ram exposure, during transition from anestrus to breeding activity, on reproductive performance. Treatments were: 1) ewes joined with two mature vasectomized Rambouillet rams for 15 d before breeding (DC), 2) ewes maintained across a net wire fence from two vasectomized rams for 15 d before breeding (FC) and 3) ewes maintained approximately 400 m away from rams (NC). At the end of the 15 d, all ewes were placed in one pasture and mated to three fertile Suffolk rams during a 34-d breeding season. A total of 96 Suffolk and 177 Suffolk-cross ewes was utilized during the 3-yr experiment. A greater (P less than .05) prebreeding ovulation percentage was observed in DC and FC than in NC ewes. Mating and lambing occurred approximately 6 d earlier for DC or FC ewes than for NC ewes. A similarly designed experiment was conducted using Suffolk and Suffolk-cross ewe lambs allotted to treatments within breed and year to measure effects of ram exposure during the natural breeding season, but prior to breeding. Treatment differences were not detected (P greater than .05) for date of first observed estrus, date of lambing, percentage of ewes lambing in the first 17 d of the lambing season, number of lambs born per ewe lamb exposed or number of lambs born per ewe lamb giving birth.

Naloxone-reversible inhibition of luteinizing hormone in postpartum ewes: effects of suckling and season.

Involvement of endogenous opioids in inhibition of luteinizing hormone (LH) release and stimulation of prolactin (PRL) release was investigated by injecting the opioid antagonist naloxone into 18 ewes on d 7 and 8, d 12 and 13, and d 18 and 19 postpartum. Compared with control injections of saline, iv naloxone (1 mg/kg) increased serum concentrations of LH and decreased serum PRL in samples collected 15, 30 and 45 min after each injection. Ewes lambing in the spring (March) or autumn (September and October) that nursed one or two lambs did not differ in their LH and PRL responses to naloxone. Autumn-lambing ewes from which lambs were weaned within 1 d after parturition did not differ from ewes of the autumn-nursed group in any of the following characteristics: 1) serum LH increases following naloxone, 2) basal secretion of LH, 3) postpartum interval to first increase in serum progesterone and 4) relative decrease in serum PRL after naloxone despite large differences in basal PRL secretion. In summary, postpartum expression of a naloxone-reversible inhibition of LH release and stimulation of PRL secretion did not depend on suckling stimuli or differ between autumn and spring parturitions.

Hypertonic saline resuscitates dogs in endotoxin shock.

In this experiment we determined if infusions of hypertonic saline (HS, 1080 Na meq/liter) could resuscitate dogs in endotoxin shock as effectively as Ringer's lactate (RL, 130 Na meq/liter). Anesthetized dogs received iv 0.5 mg/kg of Escherichia coli endotoxin, and mean arterial pressure (MAP, mm Hg) decreased from 148 +/- 5 to 58 +/- 14 within 30 min. To resuscitate the dogs 13 meq/kg of sodium was intravenously infused over 90 min as either a 10% body weight load of RL (n = 5) or a 1.2% body weight load of HS (n = 5). Both solutions produced an equivalent hemodynamic resuscitation 3 to 4 hr postinfusion with an increase in MAP (RL, 119 +/- 4; HS, 108 +/- 7), the restoration of cardiac outputs to baseline (RL, 2.0 +/- 0.2; HS, 1.9 +/- 0.3 liter/min), and similar renal inulin clearances (RL, 48 +/- 16; HS, 44 +/- 7 ml/min). The net fluid gain (resuscitation fluid volume infused minus urine output as percentage of body weight) was much greater in the RL group (7.2 +/- 1.0%) than in the HS group (0.48 +/- 0.2%). Plasma volume (PV, percentage of body weight) was measured with Evans blue dye in these splenectomized dogs. The increase in PV in the RL dogs (1.25 +/- 0.04%) was slightly greater than the increase in the HS group (0.94 +/- 0.13). Prenodal skin lymph was collected from both hindpaws, and the fractional increase in skin lymph flow after RL (4.5 +/- 2.9) was greater than the increase in the HS group (1.7 +/- 0.3).(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of estradiol-17 beta, naloxone and gonadotropin releasing hormone on postpartum secretion of luteinizing hormone in fall-lambing ewes.

The interaction among exogenous estradiol-17 beta, naloxone and gonadotropin releasing hormone (GnRH) in the control of luteinizing hormone (LH) secretion was studied in intact postpartum ewes nursing their offspring. One-half of 30 fall-lambing ewes were implanted subcutaneously with an estradiol-17 beta containing Silastic capsule between postpartum d 1 and 12 which doubled their serum concentrations of estradiol (16.0 +/- .1 vs 8.4 +/- .1 pg/ml). Blood samples were collected from implanted and non-implanted ewes at 15-min intervals for 5 h on d 3, 8, 13, 20 and 28 postpartum. Pre-injection samples were collected for 1 h, and ewes were injected with saline, naloxone (NAL;1 mg/kg) or GnRH (100 micrograms/ewe). When averaged across all days and implant groups, serum LH in the three post-NAL samples was higher (P less than .05) than in the three pre-NAL samples (3.6 +/- 1.2 vs .6 +/- .2 ng/ml). Post-GnRH concentrations of serum LH were lower (P less than .05) in estradiol-implanted ewes than in non-implanted ewes on d 8 and 13, but there were no differences in any LH characteristics on d 20 and 28 after implant removal on d 12. In non-implanted ewes, serum LH responses to GnRH increased (P less than .05) eightfold from d 3 (3.8 +/- 1.4 ng/ml) to d 8 (31.6 +/- 1.4 ng/ml), remained elevated through d 20, but declined by d 28 (10.8 +/- 1.4 ng/ml).(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of feeding time and diet on time of parturition in multiparous ewes.

Three hundred three pregnant ewes were randomly allotted to treatments in a 2 (diet) X 2 (feeding time) factorial arrangement 10 d before the first lambs were due in two lambing seasons to study the effect of time of feeding and diet on time of lambing. Behavioral activities were observed to determine if there was a relationship to time of parturition. All ewes were fed .5 kg ground whole corn per day plus either: alfalfa hay fed at 2200 each day, alfalfa hay fed at 1000 each day, alfalfa haylage fed at 2200 each day or alfalfa haylage fed at 1000 each day. Lambing data from 1983 and 1984 were combined across years because no treatment X year interactions (P greater than .05) were detected. Average time of day for lambing did not differ (P greater than .05) among treatments. In general, no differences due to treatments were observed in lambing time or behavioral activities studied. When data were pooled across treatments, two peak lambing times were observed. These peak times for lambing were from 0300 to 0700 (22.1%) and from 1500 to 1900 (22.5%). During these two 4-h periods (8 h total) 44.6% of the ewes gave birth.

Endogenous opioid modulation of luteinizing hormone and prolactin secretion in postpartum ewes and cows.

A possible role for endogenous opioid peptides (EOP) in the control of luteinizing hormone (LH) and prolactin (PRL) secretion was studied by injecting the opioid antagonist, naloxone (NAL), into postpartum ewes and cows. Twelve ewes that lambed during the fall breeding season and nursed their lambs were injected iv with NAL (1.0 mg/kg) on d 10, 14, 18, 22 and 26 postpartum. Blood samples were collected at 15-min intervals from 2 h before to 2 h after NAL, and serum concentrations of LH and PRL were quantified. Following treatment on d 10, suckling lambs were removed from 6 of the 12 ewes, creating non-suckled (NS) and suckled (S) treatment groups for subsequent study on d 14 through 26. On d 10, NAL treatment increased LH (P less than .01) but concentrations of PRL were not affected. When averaged across d 14 to 26, post-NAL concentrations of LH were greater (P less than .001) than pre-NAL concentrations (6.5 +/- .7 vs 1.9 +/- .4 ng/ml). In contrast, concentrations of PRL in the post-NAL period were lower (P less than .001) than pre-NAL concentrations (129 +/- 15 vs 89 +/- 10 ng/ml). Compared with S ewes over d 14 to 26, those in the NS group had similar pre-NAL concentrations of LH, tendencies for higher (P less than .10) post-NAL concentrations of LH, lower (P less than .001) mean serum concentrations of PRL (pre- and post-NAL) and similar pre-NAL vs post-NAL differences in serum PRL. Six suckled beef cows on d 24 to 35 were injected iv with either saline or NAL (.5 mg/kg) in a replicated crossover design. Injections of NAL increased serum concentrations of LH (P less than .05), when averaged over all 12 injections in the six cows, but serum PRL was not changed. However, three of six cows did not respond to NAL with increases in serum LH. These non-responding cows were similar to the responding cows in their pre-injection concentrations of LH and PRL, but they tended (P = .10) to have higher serum concentrations of cortisol than responding cows.

Effect of sidestream and mainstream smoke exposure on in vitro interferon-alpha/beta production by L-929 cells.

A peristaltic pump smoking machine that allows simultaneous generation of mainstream (active) and sidestream (passive) smoke from a cigarette was used to expose cultures of murine L-929 cells, a potent producer of interferon, to smoke. The cigarette used was the University of Kentucky 2R1 reference cigarette. The dosages of smoke used for exposure were the highest doses possible that generated a minimum toxic effect, and they were serially diluted to lower doses. Dosages were determined by the number of smoke puffs generated, the volume of smoke puffs generated, and the total particulate matter deposited on Cambridge filters in the smoke machine. Viability of exposed cells was equivalent to control cell cultures. Interferon-alpha/beta was induced by addition of polyriboinosinic-polyribocytidylic acid to the cells. Interferon production was substantially reduced in viable cells exposed to mainstream or sidestream smoke. Aging of smoke by delaying time of exposure of the cells to the smoke, or filtration of smoke through activated charcoal substantially decreased the alteration of interferon production by smoke exposure. These results suggest that actual exposure of cells to mainstream or sidestream smoke can inhibit in vitro interferon-alpha/beta induction, but the cells can be protected from these effects by smoke manipulation.

Consumption of nematodes by fungivorous mites, Tyrophagus spp. (Acarina: Astigmata: Acaridae).

Free-living nematodes may be attacked and eaten by soil mites which are normally considered fungivores or saprophages. Three species in the genus Tyrophagus, common inhabitants of grassland soils and also common pests in stored products, museums and laboratories, are predators of nematodes. All active stages of the mites will voraciously consume nematodes. When offered nematodes and a choice of other food (baker's yeast and algae), 11% of the Tyrophagus putrescentiae, 23% of the T. zachvatkini, and 56% of the T. similis tested fed on nematodes. Tyrophagus zechvatkini and similis were reared on a diet consisting entirely of nematodes, and developed at rates similar to a fungal diet and produced viable offspring. Agar cultures of Aphelenchus avenae which were inoculated with five mating pairs of Tyrophagus zachvatkini had populations that were one-third less than mite-free controls. Observations indicate that nematodes may be attacked by tyrophagid mites when in a dry, anhydrobiotic state.

The effect of smoke generation and manipulation variables on the cytotoxicity of mainstream and sidestream cigarette smoke to monolayer cultures of L-929 cells.

A system for the simultaneous exposure of monolayer cell cultures to mainstream (MS) and sidestream (SS) smoke from the same cigarette was utilized to study the effects of smoke generation and manipulation variables on the cytotoxicity of smoke to monolayer cultures of mouse fibroblast-like L-929 cells. The cytotoxicity of MS smoke was decreased with increasing smoke age (up to 8.7 s), smoke dilution, and the quantity of activated charcoal in filters. Acetate filters had little effect on cell mortality, and the age-of-smoke effect was not evident for MS smoke generated with a low puff volume and rapid dilution. The cytotoxicity of SS smoke also decreased rapidly with increasing smoke age and dilution. The results indicate that the gas phase of smoke may be of major importance in generating the observed toxic effects. These results may be of potential future significance in defining the requirements for a less toxic cigarette, in considering the hazards of SS smoke, and in evaluating in vivo inhalation studies.