Ozone-induced chemiluminescence of organic compounds.

Chemiluminescence was observed when organic compounds in a dry state,as well as in organic solvents, were exposed to ozone in oxygen. The peak intensity of the light emission is related quantitatively to the amount of compound. Many compounds can be identified by this method, and some are listed. Sensitivity of detection approximates that of fluorescence methods and extends to less-than- nanogram quantities for some compounds. Ozoneinduced chemiluminescence of compounds in acetone solution was less sensitive than it was in the dry analysis.

Countercurrent chromatography with flow-through coil planet centrifuge.

We have developed a new method of countercurrent chromatography which employs a vertical helical tube in the centrifugal field. The helical tube is arranged so that it does not rotate as it revolves, thus eliminating the need for rotating seals. When the gyrating tube is filled with either phase and the other phase is introduced into the tube in the proper direction, an equilibrium state results in which the two phases are split into multiple alternating segments within the coil. Each phase oscillates to and fro with the rotation as the moving phase is steadily eluted out through the other end of the tube. Consequently, solutes introduced into the tube are subjected to a rapid partition process, resulting in an efficient chromatographic separation without the complications arising from solid supports. The method is illustrated by the microanalytical separation of dinitrophenyl amino acids and can be used on a preparative scale.

Countercurrent chromatography: liquid-liquid partition chromatography without solid support.

The liquid-liquid partition chromatographic system reported here involves a long helix of narrow-bore tubing. When the coiled tube is filled with one phase of a two-phase system and fed with the other phase, phase-interchange takes place in each turn of the coil, leaving a segment of the former phase as the stationary phase. Consequently, solutes present in either phase are subjected to a multistep partition process. The column efficiency, estimated on a separation of dinitrophenyl amino acids, is comparable to that of gas chromatography.

Application of the elution centrifuge to separation of polynucleotides with the use of polymer phase systems.

A flow-through centrifuge without rotating seals enables the use of polymer phase systems for chromatographic separation of macromolecules. The capability of the technique is demonstrated on partition of polynucleotides.

Capillary-tube scanner for mechanized microbiology. A photoelectric scanner measures growth in agar-filled capillaries and gives a new approach to microbiology.

Growing microcolonies of bacteria can be detected by their light-scattering property. We have described a system in which growing bacteria in glass capillaries filled with nutrient agar are counted as the capillaries move through a narrow beam of light. Increased counts after incubation indicate the presence of viable organisms in the original sample; the equipment can detect colonies when they grow to a diameter of 8 microm. Aerobic, anaerobic, and facultative organisms can grow in the capillary environment. The sensitivities of several organisms to antibiotics were determined by adding antibiotics to the agar before the capillaries were filled.

Multicapillary mixer of solutions.

A mixer made from a bundle of glass tubules can mix two solutions within 30 microseconds, with a total-solution flow rate of 1.33 milliliters per second. One solution passes through the interstices of the bundle; the other moves through the lumens of the tubes.

New flow-through centrifuge without rotating seals applied to plasmapheresis.

The flow-through centrifuge eliminates complications arising from rotating seals. Preliminary studies on plasmapheresis demonstrated negligible platelet injury and no evidence of hemolysis during 12 hours of operation. Thus the system may provide a broad application to cell washing and elutriation, zonal centrifugation, and countercurrent chromatography.

Capillary tube scanning applied to cell growth kinetics.

We have developed an optical scanning instrument which may become a valuable aid in a variety of cell studies. Cell multiplication can be readily monitored with the instrument. By varying the chemical environment in which the cells are tested, questions of research and clinical interest may be quickly and conveniently answered. It appears possible to investigate the chemotherapeutic value of experimental agents, and to scan the sensitivity of a given population of neoplastic cells to a variety of chemotherapeutic agents at several concentrations. The optical scanning system may also be used to investigate certain metabolic pathways of cells in tissue culture, to determine drug resistance, and to assess the effects of medium composition on cell growth. Our studies of BrdU resistance in mouse lymphoma cells were done primarily to demonstrate the potential of this optical scanning method, and we do not intend to continue the cytokinetic studies at present. We trust that others will be stimulated to investigate further applications of this instrumental approach to problems in cell biology.

Droplet countercurrent chromatography.

A new form of countercurrent chromatography, named droplet counter-current chromatography, has been developed. This all-liquid separation technique is based on the partitioning of solute between a steady stream of droplets of moving phase and a column of surrounding stationary liquid phase. Milligram quantities of dinitrophenyl (DNP) amino acids were separated with an efficiency comparable to that of gas chromatography.

Artificial placenta: two days of total extrauterine support of the isolated premature lamb fetus.

A premature lamb fetus was totally sustained by extracorporeal perfusion with the use of a silicone-membrane blood oxygenator and parenteral nutritional support. The fetus remained in a metabolically stable state lasting several days.

Evidence that the brain participates in the humoral natriuretic mechanism of blood volume expansion in the dog.

We examined the role of the central nervous system in the activation of the humoral natriuretic mechanism elicited by blood volume expansion. Studies were performed in anesthetized dogs pretreated with deoxycorticosterone acetate (15 mg/day) and sodium chloride for 12 days. An isolated dog kidney perfused with blood from the femoral artery of the volume expanded dog served as the bioassay system for the humoral natriuretic factor. In group I volume expansion of intact dogs (n = 14) with equilibrated blood promoted an increase in fractional sodium excretion (FE(Na)) from a control level of 2.6+/-0.5 to 13.6+/-1.6%, P <0.001. In the isolated kidney FE(Na) increased from 3.6+/-0.8 to 6.8+/-1.1%, P <0.01. The natriuresis from the isolated kidney occurred in the absence of significant changes in renal arterial pressure, glomerular filtration rate, plasma protein concentration, or packed cell volume, whereas renal blood flow decreased slightly. In group II (n = 20) the dogs were decapitated by means of a specially designed neck vise. In 10 dogs blood pressure was supported by a constant infusion of dopamine (3.8+/-0.7 mug/min per kg body weight). Despite the fact that in response to the same volume stimulus, decapitated dogs manifested an increase in blood volume and cardiac output similar in magnitude to that of intact dogs whereas the rise in mean arterial pressure of decapitated dogs exceeded that of intact dogs, the natriuretic response of decapitated dogs was significantly less than that of intact dogs. FE(Na) in decapitated dogs increased 4.7+/-1.1 compared to 11.1+/-1.4% in intact dogs (p <0.01). Furthermore, volume expansion of decapitated dogs failed to elicit a natriuretic response from the isolated kidney. FE(Na) in the isolated kidney measured 2.6+/-0.4 before and 2.6+/-0.4% after blood volume expansion. These data indicate that decapitation inhibits activation of the humoral natriuretic mechanism elicited by blood volume expansion and are consistent with the interpretation that the brain is the source of the natriuretic factor or that the brain participates in the activation of the humoral natriuretic mechanism at some other site in the body.

Long-term survival of normal adult human tissues as xenografts in congenitally athymic nude mice.

Bronchi, pancreatic ducts, and colons from adult human were maintained as xenografts in congenitally athymic nude N:NIH(S) mice for 715, 145, and 89 days, respectively. After an ischemic crisis and revascularization of the human tissue, the epithelium regenerated to a normal differentiated morphology. The long-term survival of normal adult human epithelial tissues as xenografts provides model systems for the study of the interactions of chemical and/or physical carcinogens with human tissues.

Intra-blood-brain barrier synthesis of IgG: comparison of IgG synthesis formulas in a computer model and in 1,629 consecutive specimens.

We evaluated four formulas for estimating the intra-blood-brain barrier (IBBB) synthesis of IgG in a computer model of two types of BBB damage and in 1,629 consecutively received pairs of CSF and serum. These four formulas were the IgG synthesis rate (SR), IgG(loc) (ie, local), IgG index, and IgG extended index. Results from the 1,629 specimens and the computer model of type 2 BBB damage (partial or total loss of selective filtration) accurately predicted problems encountered with the IgG SR in clinical practice. In both the model and with the results from the 1,629 specimens, the combination of the IgG(loc) and IgG index formulas minimized the possibility of false-positive and false-negative results and yielded the most reliable results for IBBB synthesis of IgG in the presence or absence of increased permeability of the BBB. Additional studies with detailed clinical correlations will allow an accurate assessment of how effectively this combination of formulas can help to rule in or out the diagnosis of a particular disease or inflammatory condition of the CNS.

Electrical thermal angioplasty: catheter design features, in vitro tissue ablation studies and in vivo experimental findings.

A prototype electrical thermal tip catheter has been developed for ablation of atherosclerotic plaques. The operating principle is based on establishing an electric arc between a central electrode and the inside of an enclosing metallic tip to convert electrical energy to thermal energy. The metal tip catheter can be heated rapidly and boils water in a saline bath with less than 1 W of power. In vitro studies, using human atherosclerotic aortic segments, demonstrated that effective tissue ablation required more power in saline than in air (more than 9 W vs less than 1 W). The rate of tissue ablation varied with the amount of power delivered and the catheter tip pressure against tissue. In a saline field, 2-mm thickness aortic wall segments were consistently perforated in 3 seconds when 15 W of power and 20 g/mm2 of pressure were applied. Using an atherosclerotic rabbit, in vivo electrical thermal angioplasty (3 W of power for 10-second treatment sequences) was performed in obstructed iliofemoral vessels after mechanical recanalization was attempted. Of the 7 vessels that were more than 50% narrowed (compared with contiguous normal vessel diameter), mechanical recanalization was effective in only 1, but thermal angioplasty resulting in an increased lumen diameter was successful in 6 others. The recanalized segments were 8 to 10 cm; gross morphologic and histologic examination of recanalized vessels demonstrated little or no thermal injury. In addition, significant cholesterol-laden plaques remained in the vessel lumen, which appeared to have been compressed and displaced laterally.(ABSTRACT TRUNCATED AT 250 WORDS)

Multicentric papillary-cystic neoplasm of the pancreas.

Two distinct papillary-cystic neoplasms were found in the pancreas of a young black woman. She presented to the hospital in her first trimester of pregnancy with the chief complaint of sharp right upper quadrant abdominal pain that radiated to the right shoulder. This was associated with jaundice, vomiting, and pruritus. On examination, a large, nontender, midepigastric abdominal mass was palpated. Serum liver enzyme levels were moderately to markedly elevated. An abdominal computed tomographic scan revealed a 9-cm solid and cystic mass located within the head of the pancreas with associated marked bile duct dilatation. A total pancreatectomy was performed. Gross examination of the specimen revealed two separate well-circumscribed tumors of unequal size. The larger one was found within the head of the pancreas and contained multiple hemorrhagic, cystic cavities. The smaller one, located within the tail, consisted primarily of solid tissue. Microscopic examination of both lesions revealed papillary-cystic neoplasms. To our knowledge, this is the first report of two synchronous papillary-cystic tumors of the pancreas and the first reported demonstration of the potential of this tumor for multicentricity.

Ischemic changes in fetal myocardium. An autopsy series.

It has been assumed that fetal myocardial necrosis is an uncommon event that occurs only under unusual circumstances. We studied random heart sections on 76 fetal and perinatal autopsies from a 4-year period to determine the types and frequency of histologic abnormalities that occur in fetal myocardium. Vacuolar degeneration was extremely common (43% of stillbirths, 84% of live births) but a nonspecific finding. Ischemic changes, which are typically associated with coagulation necrosis, myofiber waviness, or contraction band necrosis, were seen in 21% of stillbirths and 32% of live births. In the majority of cases with histologic evidence of ischemic change, a combination of either contraction band necrosis, coagulation necrosis, and/or myofiber waviness was identified. Only rarely was any one of the abnormalities seen as an isolated feature. In only two autopsies were the ischemic changes identified in the initial autopsy report. We conclude that the histologic changes associated with ischemia that may represent myocardial necrosis are not uncommon and are frequently overlooked at autopsy.

Value of repeat cervical smears at the time of colposcopic biopsy.

At our institution, gynecologists perform Pap smears not only as an initial screening procedure but also at the time of colposcopy and biopsy. We compared the results of initial Papanicolaou (Pap) smears with those taken at colposcopic biopsy to determine if repeating the Pap smear at the time of colposcopy contributed to patient management. We found that repeat colposcopic smears often agreed with the referral smears and biopsies. When there was disagreement, the colposcopic smear was more often a lower grade than the referral smear and correlated best with the biopsy. Both referral and colposcopic smears underestimated the severity of cervical lesions with the same frequency. We could document only 5/414 (1.2%) colposcopic smears that contributed to patient care.

High density tissue culture on microporous membranes perfused by blood: report of a new bioartificial system.

A new tissue culture system has been developed to support high density cell growth using the sheep as the host. Cellular nutrition is provided by plasma solutes as they diffuse from arterial blood across microporous polycarbonate membranes into a device attached to an arteriovenous shunt. Culture chambers are constructed with transparent polycarbonate to allow photomicroscopy of the tissue in situ. System performance is demonstrated by the high density growth of fetal sheep thymus allografts and rat soft tissue sarcoma xenografts.