Tilapia (Oreochromis niloticus) dosed with azathioprine display immune effects similar to those seen in mammals, including apoptosis.

Azathioprine, an anti-neoplastic drug and therapeutic immunosuppressant, was administered intraperitoneally at 10.0 and 50.0 mg/kg to 3-6-month-old tilapia (Oreochromis niloticus). Consistent alterations in immune cellular parameters of the blood, pronephros (hematopoietic kidney) and spleen were observed. Peripheral blood total cellularity decreased as the azathioprine dose increased, to approximately half that of the control. Differential analysis of white blood cells indicated a decline in lymphocyte number, in particular, with increased dosage of azathioprine. Pronephric total cellularity was depressed in fish receiving the 10.0 or 50.0 mg/kg dose. In contrast, both splenic weight and splenic total cellularity increased proportionately with the increase in the drug dosage. Histopathologic examination of the spleens showed normal patterns for both control and 10.0 mg/kg dose groups. At 50.0 mg/kg, spleens were characterized by marked expansion of the white pulp, although lymphocytes were rare. Melanomacrophage centers at the higher dose were also larger and more numerous than in the control group. Evaluation of splenic and pronephric leukocytes with apoptotic markers showed an increase in apoptotic cells in the pronephros with increasing drug dose. These changes in fish are consistent with those seen in humans and laboratory rodents dosed with azathioprine, suggesting that fish may be potentially useful as preliminary models for detecting immunosuppressive compounds.     

Liquid chromatographic determination of benzo[a]pyrene in total particulate matter of cigarette smoke

The benzo[a]pyrene (BaP) delivery of reference and commercially available tobacco cigarettes, as well as reference and placebo marijuana cigarettes, is determined using a sequential liquid chromatographic/liquid chromatographic procedure. The total particulate matter of sample cigarette smoke is collected using a Cambridge filter pad, which is ultrasonically extracted with acetone. The resulting extract is filtered, then fractionated using semipreparative-scale normal phase liquid chromatography (LC). Quantitative determination is achieved using analytical-scale reverse phase LC equipped with a fluorescence detector. The method is precise (+/- 10-15% relative standard deviation) and yields 85% or better BaP recovery at the ng/cig. level. A single pad may be analyzed in 8 person-hours, while a more typical lot of 12 pads (6 pads each for 2 cigarette brands) may be analyzed in 10 person-days.     

Electroencephalographic patterns of clinically normal, sedated, and tranquilized newborn foals and adult horses

To establish a clinically practical procedure for recording the equine EEG, 25 healthy adult horses and 6 newborn foals were used. Recordings were taken with the animals alert and tranquilized, confined in metal stocks, or physically restrained. The dominant alert waveforms of adult horses were fast activity (25 to 40 Hz) with medium-to-low voltages (5 to 40 microV-dominant 10 to 15 microV). Underlying this fast activity was slower (0.5 to 4.0 Hz) activity with medium-to-low voltages (10 to 40 microV). Twelve of the 25 adult horses had EEG frequencies in the alpha frequency range (10 to 15 Hz, 10 to 50 microV). Eight horses were given xylazine and 17 were given acetylpromazine. Those given xylazine had generalized slowing with several distinct frequency patterns (25 to 40 Hz, 5 to 30 microV; 10 to 15 Hz, 10 to 80 microV; and 0.5 to 4.0 Hz, 10 to 90 microV). Horses given acetylpromazine had fast activity (25 to 40 Hz) with medium-to-low voltages (5 to 40 microV). Underlying this activity were slower waveforms (1 to 4 Hz) with medium-to-low voltages (5 to 10 microV). Occasional well-formed spindle activity was observed (10 to 14 Hz, 10 to 50 microV). Acetylpromazine had little effect on the EEG recording, whereas xylazine exerted a substantial effect. All leads were synchronous with lower voltages in the left frontal, right frontal, and transfrontal leads. The alert pattern of a newborn foal was characterized by low frequency (2 to 6 Hz) with medium-to-high voltages (20 to 90 microV).(ABSTRACT TRUNCATED AT 250 WORDS)     

Metal chlorides in ionic liquid solvents convert sugars to 5-hydroxymethylfurfural

Replacing petroleum feedstocks by biomass requires efficient methods to convert carbohydrates to a variety of chemical compounds. We report the catalytic conversion of sugars giving high yield to 5-hydroxymethylfurfural (HMF), a versatile intermediate. Metal halides in 1-alkyl-3-methylimidazolium chloride are catalysts, among which chromium (II) chloride is found to be uniquely effective, leading to the conversion of glucose to HMF with a yield near 70%. A wide range of metal halides is found to catalyze the conversion of fructose to HMF. Only a negligible amount of levulinic acid is formed in these reactions.     

Evaluation of an indirect enzyme-linked immunosorbent assay for screening antibody against aflatoxins

Enzyme-linked immunosorbent assay screening of antibody produced against aflatoxin was accomplished by a new and simple procedure. To demonstrate the new indirect ELISA technique used, antibody against aflatoxin M1 was produced in female BALB/CJ mice by immunization with an aflatoxin M1-bovine serum albumin conjugate. Instead of coating test-plate wells with purified antibody (direct ELISA) or synthesizing a second protein-aflatoxin conjugate (aflatoxin M1-poly-L-lysine) to coat test-plate wells, wells were coated with the readily available aflatoxin M1-bovine serum albumin and aflatoxin B1-bovine serum albumin. This method, applicable for any aflatoxin conjugated by the common cyclopentano-carboxymethoxyl-oxime technique, eliminates the more time-consuming and technically difficult portions of earlier direct and indirect ELISA. The new technique can be valuable in continued efforts toward development of new and improved immunoassays against aflatoxin metabolites.     

Technical note: preservation of tissues and gastrointestinal tract portions by plastic coating or plastination.

Two methods to preserve gastrointestinal tract (GIT) organs and tissues, plastic coating (PC) and plastination (PN), were investigated and compared. Specimens to be preserved were removed from animals within 2 h of death and immediately cleaned with water. Digesta contents were removed by flushing desired portions of GIT with water until the exiting water was clear. In the PC method, cleaned specimens were dehydrated by immersion in an isopropanol solution, dried with forced air after positioning and orientation as in situ, and finally coated on the outer and inner surfaces with a clear plastic material. In the PN procedure, specimens were filled with, and submerged in, a low-formaldehyde fixative, then dehydrated by immersion in a cold acetone solution. Dehydrated specimens were immersed in silicone and placed in a freeze drier for impregnation under low vacuum, followed by overnight gas curing with a silicone crosslinker. Finally, viewing windows were cut out with a scalpel in GIT preserved by both methods. Preserved GIT and tissues had an appearance similar to their appearance in vivo. The PC method was simple and inexpensive. Plastinated specimens were more flexible, durable, and lifelike than those preserved by the PC method. In addition, many body parts, such as muscles, nerves, bones, ligaments, and central nervous system specimens, were preserved by PN. Both methods were found to be useful tools for postmortem studies of tissues and GIT organs.     

IMAGING DIAGNOSIS—RADIOGRAPHIC,ULTRASONOGRAPHIC, COMPUTED TOMOGRAPHIC,AND FLUOROSCOPIC APPEARANCE OF A DISTAL PELVIC LIMB ARTERIOVENOUS MALFORMATION IN A YOUNG GERMAN SHEPHERD DOG

A German shepherd puppy presented for evaluation of a suspected arteriovenous fistula on the distal aspect of the right pelvic limb. Radiographs demonstrated expansion and resorption of the tarsal and metatarsal bones, and ultrasound detected a vascular abnormality. Using computed tomographic angiography, a complex arteriovenous malformation (AVM) involving the distal tibia, tarsus, and the metatarsus and an osteochondritis dissecans (OCD) lesion of the talus were identified. Based on these findings, therapeutic limb amputation was performed. Fluoroscopic angiography, vascular casting, and dissection were then used to further characterize features of this previously unreported AVM with concurrent bony lesions and OCD.     

An Additional Bone in the Carpal Region of Raptorial Birds

In a series of 4 species of owls and 12 species of diurnal raptors, an additional bone was present bilaterally in the area of the carpus. Designating the bone as an “additional bone of the raptorial carpal region” is suggested pending appropriate embryological studies to determine the origin of the bone. Generally, the bone was present in owls and in diurnal raptors with relatively short, rounded wings, but was absent in species with longer, more tapered wings. The bone lies at the cranial edge of the carpal area near the radial carpal bone, and is fixed to the surrounding skeleton by ligaments. The tendon of the tensor patagium longus muscle has one slip that attaches to the additional bone and another that passes over its craniodorsal surface en route to insertion. Awareness of the occurrence of this bone in some raptors, but not others, is essential in clinical evaluation of wing injuries, as well as in forming a prognosis for the return to soundness, the potential for release, and, therefore, the life of an injured raptor.