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The aim of this study was to evaluate the effect of topical 1% tetracaine hydrochloride on the intraocular pressure (IOP) in ophthalmologically normal horses. Thirty eyes of 15 clinically normal horses were used for this study. The animals were randomly assigned to two groups (treatment and control). Prior to the instillation of 1% tetracaine or placebo, the baseline IOPs (T0) of each animal were recorded in both groups. Then one drop of tetracaine was instilled randomly into one eye of each horse in the treatment group (8 horses). In the control group (7 horses), one drop of artificial tear was instilled in one randomly selected eye. The measurements were repeated at 2 minutes (T2), 5 minutes (T5), 15 minutes (T15), and 30 minutes (T30) post instillation via a rebound tonometer. There was no significant difference in the treatment group (P = .3). The peak IOP measured at T2 returned to the baseline value at T30. No significant difference was found in the mean IOP values between the treatment and the control groups, or between the males and females on any of the occasions (P > .05). The Results of this study revealed a nonsignificant increase of the IOP 2 minutes post instillation of 1% tetracaine in horses., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Objective: Frailty is a multidimensional vulnerability due to age associated decline. The impact of frailty on long term outcomes was assessed in a cohort of vascular surgical patients., Methods: Patients aged over 65 years with a length of stay greater than two days admitted to a tertiary vascular unit over a single calendar year were included. Demographics, mode of admission, and diagnosis were recorded alongside a variety of frailty specific characteristics. Using the previously developed Addenbrookes Vascular Frailty Score (AVFS - 6 point score: anaemia on admission, lack of independent mobility, polypharmacy, Waterlow score > 13, depression, and emergency admission) the effect of frailty on five year mortality and re-admission rates was assessed using multivariable regression techniques. The AVFS was further refined to assess longer term outcomes., Results: In total, 410 patients (median age 77 years) were included and followed up until death or five years from the index admission. One hundred and thirty-four were treated for aortic aneurysm, 75 and 96 for acute and chronic limb ischaemia respectively, 52 for carotid disease, and 53 for other pathologies. The in hospital mortality rate was 3.6%. The one, three, and five year survival rates were 83%, 70% and 59%; and the one, three, and five year re-admission free survival rates were 47%, 29%, and 22% respectively. Independent predictors of five year mortality were age, lack of independent mobility, high Charlson score, polypharmacy, evidence of malnutrition, and emergency admission (p < .010 for all). Patients with AVFS 0 or 1 had restricted mean survival times which were one year longer than those with AVFS 2 or 3 (p < .001), who in turn had restricted mean survival times over one year longer than those with AVFS of 4 or more (p < .001)., Conclusion: Frailty factors are strong predictors of long term outcomes in vascular surgery. Further prospective studies are warranted to investigate its utility in clinical decision making., (Copyright © 2020 European Society for Vascular Surgery. Published by Elsevier B.V. All rights reserved.)

Objective: To measure intraocular pressure (IOP) in horses during hoisting after induction of anesthesia., Study Design: Prospective nonrandomized clinical study., Animals: Eighteen healthy adult horses aged [mean±standard deviation (SD)] 10±4.2 years and weighing 491±110 kg anesthetized for elective procedures., Methods: IOP was measured in the superior eye of each horse based on planned recumbency after induction of anesthesia. Measurements were taken directly after premedication with xylazine or detomidine with butorphanol, after induction with diazepam-ketamine, after intubation, when suspended by the hoist and on the operating table. During hoisting, the head was supported and the eye-heart height was measured to account for variations in head positioning among patients. IOPs were compared across time points using repeated-measures analysis of variance. Regression was used to compare IOP outcome with potential cofactors., Results: Compared with measurements after premedication (17.5±2.5 mmHg) (mean±SD), hoisting significantly increased IOP (32.4±15.3 mmHg) (p<0.01). The highest recorded IOP in the hoist was 80.0 (range, 16.0-80.0) mmHg. The difference in IOP between premedication and hoisting was 15.0±16.2 (range, -1.0 to 68.0) mmHg. Body weight had a significant effect on absolute IOP and change in IOP in the hoist (p<0.01)., Conclusions and Clinical Relevance: Hoist IOP was significantly higher than post-premedication IOP with heavier horses having higher hoist IOPs and greater increases in IOP. The clinician should take this relationship into account when anesthetizing and hoisting larger horses where an increase in IOP could be detrimental., (Copyright © 2017 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.)

Unlabelled: Wound dressings are a key component in provision of optimal conditions for bleeding control and wound healing. For absorbent dressings, electrostatic interactions are frequently utilized as one of the mechanisms driving dressing adhesion. Herein, a choline phosphate functionalized biocompatible cellulose membrane that can efficiently arrest human red blood cells was developed to have potential application in wound dressing. The bioadhesion is based on the unique multivalent electrostatic interaction between the head groups of phosphatidyl choline based lipids on the cell membrane and its inverse orientation but virtually identical structure, choline phosphate, coupled to the cellulose membrane. For functionalization, the cellulose membrane was decorated with polymer brushes bearing multiple choline phosphate groups via surface-initiator atom transfer radical polymerization followed by click chemistry. The modified cellulose membranes were characterized by ATR-FTIR and the molecular weight and the grafting density of polymer brushes grafted from the cellulose membrane surface were thoroughly evaluated by calibrated force-distance measurements with atomic force microscopy (AFM). This new method provides an approach to estimating polymer brush parameters on rough surfaces of unknown surface area based on the dependence of brush thickness on brush density and polymer molecular weight for a calibration set of brushes. The dependence of binding of human red blood cells (RBCs) to the cellulose membrane surface on the number density of choline phosphate groups (e.g. molecular weight) and the grafting density were investigated using this AFM-based approach. Bound RBCs showed "pseudopodia"-like membrane projections under scanning electron microscopy where cells contacted the microfibers of the cellulose, distorting the RBC shape, reflecting the multivalent interactions between the RBCs and the choline phosphate-doped cellulose membrane. We believe this efficient strategy provides a promising approach to blood conservation and trauma management., Statement of Significance: Uncontrolled bleeding can dramatically affect morbidity and mortality. Absorptive wound dressings provide either adherent or non-adherent layers to control bleeding. Our new adherent material is based on a universal adhesion reaction between cell membrane phosphatidyl choline (PC) headgroups and cellulose membranes (CM) decorated with polymer brushes carrying a CP group per monomer. The CP-PC multivalent interactions provide adherence to cut tissue margins and blood cells, blocking bleeding. We here demonstrate the strong specific binding of red cells to CM-CP but not CM-PC membranes and determine the requisite brush molecular weight and surface concentration via a new approach using atomic force microscopy, applicable to rough surfaces. We believe this strategy provides a promising approach to blood conservation and trauma management., (Copyright © 2016. Published by Elsevier Ltd.)

Background: There are no guidelines for the management of accidental insulin administration. We hypothesized that home monitoring of asymptomatic patients (pts) was safe following unintentionally insulin administration., Methods: Retrospective review of poison center (PCC) charts from 1/1/2000-12/31/2010 looking for accidental insulin administrations., Inclusion Criteria: pt must be prescribed insulin. Information recorded from charts: pt age/gender, "intended" and "mistaken" insulin formulations/doses, use of oral diabetic agents, management site, Emergency Department (ED) referral, symptoms, blood glucose values, and treatments. Defined outcomes: symptoms (e.g., altered sensorium); hypoglycemia (<60 mg/dL); management site; health care facility (HCF) admission; and death. Multiple logistic regression was used to determine outcome predictors., Results: 652 charts met inclusion criteria. Mean age was 56.4 years; most (58.5%) were women. Most (89%) calls originated from home, 10.7% from a HCF, 0.3% from Emergency Medical Services (EMS). Overall, 397 (60.9%) pts were managed at home. Two pts managed at home were later evaluated by EMS; neither required admission. Symptoms developed in 56 (8.6%) pts. There were no deaths. Only 40 (6.1%) pts were admitted to a HCF; 18 (45%) pts were hypoglycemic. The development of hypoglycemia (odds ratio [OR] 5.94; p < 0.001) and amount of insulin accidentally administered (OR 1.04; p < 0.001) predicted HCF referral. The type and dose of insulin administered did not predict symptoms., Conclusions: Based on a retrospective analysis of a single PCC's cases, home observation of asymptomatic patients after unintentional administration of a wrong insulin formulation appears safe., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Background: Fentanyl is a potent synthetic opioid with large abuse potential. A common preparation of fentanyl is a sustained-release transdermal patch. To our knowledge, there are only two published case reports of whole patch ingestion. A case series of 76 patients with a history of whole patch ingestion is reported., Study Objectives: To characterize whole fentanyl patch ingestion to develop a clinical guideline for management., Methods: This was a retrospective review of all patients who ingested intact fentanyl patches as reported to three regional poison information centers (RPIC) from 2000 to 2008. The three RPIC medical record databases were queried for all exposures with a substance code matching the Micromedex® (Thomson Reuters, New York, NY) fentanyl product codes. Collected data included: age, gender, reason for the exposure, number of patches ingested, dose (μg/h), symptoms, symptom onset and duration, treatment hospital flow (level of care), and outcome., Results: A total of 76 patients met the inclusion criteria. Two patients had both time of onset and symptom duration documented. In both patients, the signs and symptoms developed within 2 h of the exposure, and the patients were asymptomatic at 6½ and 9 h, respectively. Fifty-eight (78.3%) patients were admitted. Of those patients who were admitted, 56 (96.5%) were admitted to a critical care unit. Fourteen patients required intubation, and naloxone infusions were documented in eight cases., Conclusion: Ingestion of whole fentanyl patches may lead to prolonged and significant toxicity based on these poison center data., (Copyright © 2012 Elsevier Inc. All rights reserved.)

This is the third article of a three-part series that reviews the care of poisoned patients in the ICU. This article focuses on natural toxins, such as heavy metals and those produced by plants, mushrooms, arthropods, and snakes. The first article discussed the general approach to the patient, including laboratory testing; the second article focused on specific toxic agents, grouped into categories.

This is the second of a three-part series that reviews the generalized care of poisoned patients in the ICU. This article focuses on specific agents grouped into categories, including analgesics, anticoagulants, cardiovascular drugs, dissociative agents, carbon monoxide, cyanide, methemoglobinemia, cholinergic agents, psychoactive medications, sedative-hypnotics, amphetamine-like drugs, toxic alcohols, and withdrawal states. The first article discussed the general approach to the toxicology patient, including laboratory testing; the third article will cover natural toxins.

Poisonings, adverse drug effects, and envenomations continue to be commonly encountered. Patients often present critically ill and warrant ICU admission. Many other patients who are initially stable have the potential for rapid deterioration and require continuous cardiopulmonary and neurologic monitoring. Given the potential for rapid deterioration, and because patients need continuous monitoring, ICU admission is frequently required. This article is the first of a three-part series to be published in CHEST; it discusses general management, laboratory tests, enhanced elimination, and emerging therapies. The second article will address the management of specific overdoses; the last will cover plants, mushrooms, envenomations, and heavy metals.

Background: Acetaminophen (APAP) toxicity is commonly encountered in the Emergency Department. Until 2004, treatment consisted of either oral N-acetylcysteine (NAC) or filtered oral NAC administered intravenously (i.v.). Intravenous acetylcysteine (Acetadote) is a new Food and Drug Administration-approved i.v. formulation of acetylcysteine manufactured by Cumberland Pharmaceuticals in Nashville, Tennessee. Little post-marketing data exists on the effectiveness and safety of i.v. acetylcysteine., Objectives: We evaluated the clinical presentations and outcomes of patients treated with i.v. acetylcysteine for APAP toxicity., Methods: We performed a retrospective chart review of patients treated with i.v. acetylcysteine for APAP ingestion. The primary outcome measures were: adverse reactions to and effectiveness of i.v. acetylcysteine, as defined by elevation of transaminases, liver failure, renal failure, death, and hospital length of stay (LOS). Data collected included: comorbidities, allergies, intentionality, timing and dosing of i.v. acetylcysteine, hospital LOS, transaminases > 1000 IU/L, development of liver failure requiring transplant, development of renal failure requiring hemodialysis, death, and anaphylactoid reactions., Results: Sixty-four patients met our study criteria. Overall, 16 (25%) patients developed transaminases > 1000 IU/L, 4 (6%) of them died and 2 (3%) received liver transplants. Of the 15 patients (23%) treated within 8 h, none died or developed liver or renal failure, and only 1 developed transient transaminase elevation > 1000 IU/L. In the patients treated outside of 8 h, the median LOS was 3 days, whereas the group treated within 8 h had a median LOS of only 1 day. Six (9%) patients developed anaphylactoid reactions, 2 of whom received the i.v. acetylcysteine bolus over 15 min. Five of these patients were treated pharmacologically and completed treatment, and one had treatment discontinued for undocumented reasons., Conclusion: Intravenous acetylcysteine seemed to be a safe and effective formulation of N-acetylcysteine., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Oral methadone therapy is an effective and increasingly popular treatment for opioid dependency and chronic pain. Although it is not typically considered pro-dysrhythmic, we present the unique case of a 52-year-old HIV-positive woman without underlying cardiac disease who developed QTc prolongation and pulseless Torsades secondary to high dose methadone therapy.

Envenomation from cobra bites causes major morbidity and mortality in Asia and Africa but rarely in the United States. We describe two patients bitten by the Asiatic Cobra (Naja Kaouthia)--both successfully treated in the emergency department. Patient 1 was a 23-year-old woman bitten in the buttock by her cobra. Examination demonstrated two puncture wounds. She developed cranial neuropathy, respiratory failure, and coagulopathy 10 h later, necessitating endotracheal intubation and polyvalent antivenom administration. The patient recovered fully with minimal wound necrosis. Patient 2, a 44-year-old man, was bitten on the hand by his cobra. Examination revealed a puncture wound with progressive swelling. Edrophonium and monovalent antivenom were administered, and he recovered uneventfully. These cases emphasize the varied clinical presentations of the Asiatic cobra. Patient 1 developed delayed neurotoxicity, respiratory failure, and hematotoxicity with minimal wound necrosis, whereas Patient 2 experienced a more typical clinical course.

The purpose of this report is to describe an unusual presentation of anaphylaxis after first-time rattlesnake envenomation. A patient on a medical toxicology inpatient service is presented who had signs of anaphylaxis, including airway compromise, after first-time rattlesnake envenomation. An epinephrine drip and oral intubation were initiated. This case is unusual in that dermal and gastrointestinal exposure may have been the primary sensitization process that preceded a severe anaphylactic reaction after envenomation. The patient's recovery was prolonged. In conclusion, rattlesnake envenomation may result in rapidly progressive airway compromise, possibly caused by anaphylaxis in patients with previous dermal or gastrointestinal exposure to snake proteins.

Salicylate (SAL) produces mitochondrial membrane permeability transition (MPT) with resultant oxidative phosphorylation uncoupling. Cyclosporine A (CSA) inhibits SAL-induced MPT. This study determined if CSA pretreatment prolonged survival time in SAL-poisoned rats. Twenty-nine rats were randomized to receive pre-treatment with either 30 mg/kg CSA or equal volume of control diluent intraperitoneally (i.p.). Four hours later, all rats received 1700 mg/kg sodium salicylate i.p. Survival time, whole blood CSA ([CSA]), and serum sodium ([Na]), glucose and SAL ([SAL]) concentrations were determined. The results showed median survival time for controls was 18 min (95% CI 14-22 min) and for CSA animals was 14 min (95% CI 13-15 min). Univariate and multivariate analyses and Cox proportional hazard regression revealed CSA treatment was associated with higher [SAL], which was associated with shortened survival times. The CSA group also demonstrated shorter survival times for a given [SAL]. In conclusion, CSA pre-treatment shortened survival in SAL-poisoned rats.

Respiratory compromise after rattlesnake envenomation (RSE) is an uncommon yet potentially lethal complication. We were interested in determining the frequency of respiratory compromise in patients treated for RSE. The incidence and indications for intubation were also determined. A retrospective chart review was conducted of all patients treated by medical toxicologists at a tertiary referral hospital between July, 1994 and November, 2000. Out of 294 total patients, 289 charts were reviewed. Of all 289 patients, 214 (74%) received Crotalidae Polyvalent Antivenin (Wyeth-Ayerst) and 23 (8%) had clinical evidence of respiratory compromise. Thirteen of 289 patients (4.4%) were intubated following RSE. No one was intubated for antivenin-induced complications. There were no deaths among studied patients during acute hospitalization. Respiratory compromise following RSE is rare, occurring in only 8% of studied patients. Only 2 patients (0.7%) required intubation as a direct consequence of RSE. No one required intubation for antivenin-induced hypersensitivity reactions.

In an effort to model the interaction of lipid-based DNA delivery systems with anionic surfaces, such as a cell membrane, we have utilized microelectrophoresis to characterize how electrokinetic measurements can provide information on surface charge and binding characteristics. We have established that cationic lipids, specifically N-N-dioleoyl-N,N-dimethylammonium chloride (DODAC), incorporated into liposomes prepared with 1, 2-dioleoyl-i-glycero-3-phosphoethanolamine (DOPE) or 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) at 50 mol%, change the inherent electrophoretic mobility of anionic latex polystyrene beads. Self-assembling lipid-DNA particles (LDPs), prepared at various cationic lipid to negative DNA phosphate charge ratios, effected no changes in bead mobility when the LDP charge ratio (+/-) was equal to or less than 1. Increasing the LDP concentration in a solution of 0.1% (w/v) anionic beads resulted in a charge reversal effect when a net charge of LDP to total bead charge ratio (+/-) of 1:1 was observed. LDP formulations, utilizing either DOPE or DOPC, showed similar titration profiles with a charge reversal observed at a 1:1 net LDP to bead charge ratio (+/-). It was confirmed through centrifugation studies that the DNA in the LDP was associated with the anionic latex beads through electrostatic interactions. LDP binding, rather than the binding of dissociated cationic lipids, resulted in the observed electrophoretic mobility changes of the anionic latex beads., (Copyright 1999 Academic Press.)

Polyclonal antibodies raised against the hallucinogenic drug, lysergic acid diethylamide (LSD), were used to detect and extract drug from whole blood samples. An indirect ELISA was used to detect as little as 1 pg of total drug in 25 microl blood. The limit of detection of the immunoassay, calculated from the mean - 3 SD was 39 pg/ml. The analytical recovery of LSD (2.5-0.2 ng/ml) from whole blood was 102-113%. Within-run CVs for LSD spiked in blood at 1.25, 0.16 and 0.04 ng/ml were 5.6, 3.1, and 8.9%, respectively (n = 4). There was an overall decrease in precision when whole blood was used in place of urine, due to the increased complexity of the matrix. However, using this technique LSD was calibrated in blood in the sub-ng/ml region of forensic interest. Immunoaffinity extraction was used to isolate LSD from blood and urine samples. The affinity support was prepared by covalently attaching anti-LSD antibodies to Protein A-coated agarose beads. No pre-treatment of the sample was required other than the addition of neutral buffer. Sub-ng/ml concentrations of LSD were routinely extracted from blood and urine samples with greater than 80% recovery of drug. This technique, which could be used to extract LSD from blood and urine samples prior to confirmatory drug analysis, could be completed in about 10 min.

Complement activation causes opsonization of foreign particles leading to particle elimination from the blood. Complement-mediated opsonization of charged and large liposomes presents a fundamental problem in their use to deliver therapeutic agents in vivo. To prolong the circulation half-lives of such liposomes, complement activation must be curtailed. The aim of this study was to assess the ability of poly(ethylene glycol)-lipids (PEG-lipids) to inhibit the in vitro activation of the classical pathway of complement in human serum by anionic liposomes. Incorporation of cholesterol-PEG600 (CH-PEG600), cholesterol-PEG1000 (CH-PEG1000), or phosphatidylethanolamine-PEG2000 (PE-PEG2000) resulted in dose-dependent inhibition of C1q binding and complement activation. The dose of PEG-lipid at which complement activation was blocked was inversely related to the PEG chain length. Complement activation was strongly inhibited when 15 mole% of CH-PEG600, 10 mole% CH-PEG1000, or 5 mole% PE-PEG2000 was incorporated into 100-nm anionic liposomes. PEG-lipid incorporation into larger liposomes (240 nm) was also successful in blocking C1q binding and complement activation. Radiolabeled cholesterol-PEG approximately 1400 was prepared and used to determine both the percentage of CH-PEG incorporated into the liposomes and the percentage maintained in the liposomes in the presence of 50% human serum at 37 degrees C for up to 24 h.

Surface localized poly(ethylene glycol) (PEG) amphiphiles of type C16:0-EO151 and C18:2-EO151 were studied via ellipsometry at macroscopic, flat methylated silica (MeSi), phosphatidic acid (PA), and phosphatidylcholine (PC) surfaces. At these surfaces the amphiphiles adsorb similarly, in a non-cooperative manner, achieving a plateau (approximately 0.1 PEG chains/nm2) well below amphiphile critical micelle concentration (CMC). The resultant PEG-enriched layers were 10-15 nm thick, with a polymer concentration (approximately 0.07 g/cm3) greater than the PEG-enriched phase of many dextran, PEG aqueous two-phase systems. PEG-amphiphile adsorption (mg/m2) at hydrophobic and phospholipid flat surfaces correlated with changes in the partition (log K) of PC liposomes in such two-phase systems. PEG-amphiphile adsorption at macroscopic surfaces appears to represent a balance between hydrophobic attraction and repulsive intra-chain interactions which promote chain elongation normal to the surface.

Novel aqueous polymeric two-phase systems are described. These systems are formed by mixing hydroxypropyl cellulose (molecular mass 100,000, trade name Klucel L) with poly(ethylene glycol)-co-poly(propylene glycol) copolymer [molecular mass 6,500, poly(propylene glycol) content 50% w/w, trade name Pluronic P105], in a saline buffer. The phase diagram was measured and the interfacial tensions, phase separation times, and lower phase viscosities of three phase systems having constant Pluronic P105 concentration but varying in Klucel L concentration were determined. The partition behavior of a representative cell, bacterium, and protein and the affinity ligand-mediated alteration in the partition behavior of a protein from a yeast extract protein mixture were also characterized. The results suggest that Klucel L/Pluronic P105 phase systems may be cost-effective substitutes for, or complements to, existing aqueous polymeric phase systems. The physical characterization and representative partition data reported here should facilitate application of these new systems.

Increased intraocular pressure is accepted as a primary etiologic factor for the atrophy of the optic nerve head and visual field defects of high-tension glaucoma. Other factors must be present to explain these findings in low-tension glaucoma. One of the current theories is that low-tension glaucoma is the result of decreased optic nerve perfusion on the basis of vascular disease or other factors such as altered blood viscosity. This study compared the non-invasive vascular profiles, coagulation tests, and rheological profiles of 46 consecutive cases of low-tension glaucoma with 69 similarly unselected cases of high-tension glaucoma and 47 age-matched controls. Despite the multifactorial approach and the use of previously validated objective tests, no significant group differences were detected with any of the above investigations. If vascular disease is important in the etiology of low-tension glaucoma, then it must be localized or vasospastic since this study does not support the concept of a generalized vascular etiology, either of an atheromatous or hyperviscous nature, for the genesis of low-tension glaucoma.

The interactions of halothane with five major constituents of human blood were studied by equilibrium dialysis. Haemoglobin, albumin, red cell membranes and triglycerides were found to contribute significantly to the solubility, and thus the transport, of halothane in whole blood. At physiological concentrations, gamma-globulin had no detectable effect on the solubility of halothane. Absorption isotherms of halothane binding to haemoglobin and albumin suggest a possible positive co-operative effect, but complete saturation of the binding sites was not observed even when the aqueous phase was saturated with halothane. The number of halothane molecules bound per albumin molecule was not significantly affected by the presence of oleic acid. For red cell ghosts and triglyceride-rich micelles (chylomicrons and very low density lipoprotein), the adsorption isotherms suggested that halothane is solubilized within the hydrophobic regions. The distribution of halothane between cells and plasma calculated from the above equilibrium dialysis results was in reasonable agreement with the distribution in whole blood determined by an independent method.

A new, mild method is described for spin-labelling sialic acid residues in situ. The procedure involves the formation of C-1 sialamides and has been applied to a serum glycoprotein, a mucin, tissue sections from human colon, and erythrocyte membrane components. The selectivity of the method and its possible applicability to other types of labelling are discussed.

The value of measurements of visual evoked potential (VEP) for neurological assessment of the adult is widely accepted. Its use for neonatal evaluation following birth asphyxia has been limited by our knowledge of developmental changes occurring in the newborn brain. VEPs and electroretinograms (ERGs) were simultaneously recorded from five healthy newborn lambs from birth until 30 days of life. Newborn age was then compared with amplitude (height of the signal) and latency (time from stimulus to a specific wave deflection) measurements from these neuroelectrical signals. Latencies to A wave (photoreceptor activity) and B wave (bipolar and ganglion cells) of the ERG declined in a small but significant manner, while the latency interval from A to B wave did not change with age. Latency to wave N1 of the VEP remained stable while latency to P2 declined from birth to 30 days. The ERG and VEP signals exhibited linear increases in amplitude as the lambs became older. We conclude from these observations that ganglion and bipolar cells (B wave of the ERG) within the retina and primary nerve tracts (N1 of the VEP) along the visual system are mature at birth. In contrast, photoreceptor activity within the retina (A wave of the ERG) and nerve conduction through the visual cortex (P2 of the VEP) exhibit changes which are consistent with ongoing maturation of these more specialized areas through the early newborn period.

The possibility of producing biospecific affinity ligands for separating cells in two polymer aqueous phase systems on the basis of cell surface antigens was investigated. Rabbit anti-human erythrocyte IgG was reacted with cyanuric chloride-activated monomethyl poly(ethylene glycol) (PEG) fractions (molecular weights approximately 200, 1900, and 5000) at various molar ratios of PEG to protein lysine groups. The partition coefficient of the protein in a Dextran/PEG two-phase system increased with increasing degree of modification and increasing PEG molecular weight. There was a concomitant loss in ability to agglutinate human erythrocytes. The ability of the modified IgG to bind to a DEAE-cellulose column was almost eliminated by reaction with the PEG 5000, and was decreased to a lesser extent by PEG 1900. This PEG 1900-modified IgG substantially increased the partition of fresh or fixed human erythrocytes into the PEG-rich phase of a suitable phase system, while having no effect on rabbit cell partition. The partition increase could be inhibited by unmodified anti-human red cell IgG but not by nonspecific unmodified human IgG, demonstrating that the ligand effects were specific for the cell type against which the antibody was raised. A mixture of rabbit and human erythrocytes, which ordinarily have very similar partitions in the phase systems used, could be separated on a countercurrent distribution apparatus using the modified IgG. These results demonstrate the feasibility of producing immunologically specific affinity partition ligands for cell separation.

An alternative primary amine assay utilizing 4-phenylspiro[furan-2(3H),1'-phthalan]-3,3'-dione (fluorescamine) is compared with the frequently applied trinitrobenzenesulfonic acid assay as a means of estimating protein primary amine modification with cyanuric chloride attachment of polyethylene glycol (PEG). The results of the two assays are compared for three different proteins and several advantages of the fluorescamine assay are emphasized. In particular, the fluorescamine assay was found to be unaffected by free PEG up to 0.08% in the assay mixture. The fluorescamine assay can be used to measure the partition coefficients of native or PEG-modified protein in two-phase aqueous polymer systems.

Cell separation using aqueous polymeric two-phase systems is well established. For separations of cells having similar partition coefficients a multi-step counter-current distribution procedure has been used. As an alternative strategy, we have developed a column-based technique in which the dextran-rich phase of a dextran-polyethylene glycol (PEG) phase system is immobilized onto derivatized chromatography beads. The PEG-rich phase is used as the eluent. A separation of erythrocytes from different species utilizing an affinity ligand was performed. The following behaviour was observed: (i) no elution occurs below a critical affinity ligand concentration; (ii) cell recovery is proportional to affinity ligand concentration; (iii) elution volume is independent of affinity ligand concentration. The above suggest that no multi-step partitioning occurs. However, the separation achieved is considerably better than in a corresponding single-tube separation. A 1:1 mixture of dog and human erythrocytes was separated to a purity of greater than 98% using the column-based technique. The corresponding optimized single-tube separation gave a maximum purity of 84%.

Cell separation using aqueous polymeric two-phase systems is well established. For separations of cells having similar partition coefficients a multistep countercurrent distribution procedure has to be used. However, its operation is limited by time and apparatus constraints. As an alternative strategy we have developed a chromatographic technique in which the dextran-rich phase of a dextran/polyethylene glycol (PEG) phase system is immobilized onto derivatized agarose beads. The PEG-rich phase is used as the eluent. Inclusion of PEG-fatty acid affinity ligand gradients into the eluent produces separations of mammalian erythrocytes based on the differential interaction between the fatty acid and the erythrocyte membranes. A model separation of dog and human erythrocytes has been carried out.

Visual evoked potentials were measured in eight exteriorized near-term fetal lambs prior to and during 8 minutes of asphyxia and during 1 hour of newborn survival. Four of the lambs were then extubated, and measurements were continued while the lambs were observed for behavior and gross motor function. Early in the asphyctic period, marked changes in the visual evoked potential occurred in association with a fall in Po2, whereas during the postexposure period the visual evoked potential appeared to recover independently of cardiovascular or arterial blood gas values. Moreover, early wave components of the visual evoked potentials were more resistant to asphyxia than were later components and responded more quickly to resuscitative efforts. During the survival period the visual evoked potential did not correlate with gross motor function when animals were exposed to moderate acidosis. After severe acidosis abnormal visual evoked potentials were seen during periods of clinical deterioration.

The effect on the partition of erythrocytes in a two phase aqueous polymer system based on dextran T500 and polyethylene glycol (PEG) 8000 of a combination of immunoaffinity ligands, namely, rabbit immunoglobulin G (IgG) and PEG 1900-modified monoclonal IgG, was examined as a potential cell separation technique. Several hybridoma lines secreting mouse monoclonal IgG specific for the Fc receptor of rabbit IgG were raised. The monoclonal IgG was modified by cyanuric chloride attachment of PEG 1900, causing the modified antibody to partition predominantly into the PEG-rich upper phase of the systems. The PEG-modified monoclonal IgG was used as an affinity ligand in the two phase polymer system to specifically increase the partition of rabbit anti-NN glycophorin IgG. The rabbit IgG was applied together with the PEG-modified monoclonal IgG to increase the partition of human erythrocytes. The same system had no effect on the partition of rabbit erythrocytes. These experiments demonstrate that a monoclonal antibody can be modified and used as a general reagent with which to alter cell partition in two phase aqueous polymer systems in an immunologically specific manner.

Dacron fabrics with a wide range of porosities were autoclaved for 3 minutes after being soaked in serum, 5% albumin, or 25% albumin. Porosity of compound Dacron grafts made with 25% albumin was less than 1 ml/min/cm2 regardless of the fabric base, whereas porosity of grafts made with serum or 5% albumin was proportional to the porosity of the base fabric. Porosity of the compound grafts remained stable for more than 48 hours and to pressure greater than 450 mm Hg, if the grafts were kept moist. Tubes of Marlex mesh coated with heat-denatured albumin, implanted as infrarenal aortic replacements in dogs, showed complete albumin absorption by 3 weeks. However, perigraft tissue reaction and graft incorporation were minimal and extensive false aneurysm formation resulted. Knitted filamentous Dacron 6 mm tubes coated with heat-denatured albumin were implanted as iliofemoral bypass grafts in 12 dogs, with blood-preclotted knitted filamentous Dacron grafts implanted as contralateral control grafts. Comparison of the albumin-coated grafts with the blood-preclotted control grafts showed no differences in healing or patency at 4 to 6 months. Heat-denatured 25% albumin forms a strong and hemostatic coating regardless of fabric base. Albumin-Dacron compound grafts are easily and rapidly made in the operating room, handle well, and are suitable for large and medium-sized arterial replacements without changes in healing or patency. Because of slow tissue incorporation, however, albumin-coated knitted Dacron grafts should be avoided in patients who require long-term anticoagulation therapy.

This communication describes the design and construction of a new external injection port for the direct gas--liquid chromatographic analysis of volatile compounds in whole blood. Aliquots (4--40 microliter) of EDTA anti-coagulated blood containing the volatile compound and a weighed quantity of the internal standard, isobutanol, were injected into the disposable glass wool filter of the carrier gas stream directly onto the chromatographic column. Typical data are presented from chromatography performed with dual 6 ft. x 2 mm I.D. glass columns containing Chromosorb 101 programmed from 110--180 degrees at 6 degrees/min and the external injection port maintained at 180 degrees. The method eliminated the problems usually associated with direct injection methods and permitted the accurate analysis of halothane, methoxyflurane, diethyl ether and ethanol over the approximate range 1--100 mg%. Using this analytical procedure the distribution of halothane between the cells and plasma of human blood at 4 degrees was found to be 2.0 +/- 0.2.