Partition of gross peripheral nerve activity into single unit responses by correlation techniques.

The neural activity in the medial articular nerve is partitioned on the basis of conduction velocity by correlation techniques. Because of the small number of cat knee joint units (two to ten) responding under a prescribed set of rotational conditions, it is possible to partition and simultaneously study the units' responses to periodic joint articulation waveforms.

Stratospheric nitrogen dioxide in the vicinity of soufriere, st. Vincent.

In April 1979, measurements of nitrogen dioxide in the upper atmosphere were made near Soufriere Volcano by twilight optical-absorption techniques. The derived value of 5 x 10(15) molecules per square centimeter column implies an enhancement of 25 percent over earlier abundances measured in the same latitudinal regions. This enhancement may represent the normal stratospheric variability of nitrogen dioxide in the equatorial region but in any case may be considered an upper limit to the volcano's effect on the total nitrogen dioxide abundance.

Biosynthesis of cytidine diphosphate diglyceride by human platelets.

Homogenates of human platelets contain an enzyme which catalyzes the formation of cytidine diphosphate diglyceride from cytidine triphosphate and phosphatidic acid. The enzymatic activity could not be dissociated from platelet particles and the greatest specific activity was found in the membrane fraction. The K(m) for cytidine triphosphate was 0.16 mmole/liter and the apparent K(m) for phosphatidic acid was 6.2 mmoles/liter. The pH optimum was 7.0 and the most effective buffers were triethanolamine-HCl and Tris-HCl. The reaction was dependent on the presence of divalent cations, magnesium being the most effective of those investigated. Monovalent cations did not alter the reaction rate. Evidence is presented that the cytidine diphosphate diglyceride produced can serve as a precursor for the synthesis of phosphatidylinositol. No difference was found in the enzymatic activity in platelets from normal subjects and from patients with diseases known to interfere with platelet thromboplastic function.

The biosynthesis of phosphatidylinositol in human platelets.

Homogenates of human platelets can mediate the synthesis of phosphatidylinositol from myoinositol and cytidine diphosphate diglyceride. The cytidine diphosphate diglyceride: myoinositol, phosphatidyl transferase activity is particulate-bound, and the highest specific activity is found in the membrane fraction. The production of phosphatidylinositol is decreased by sulfhydryl-binding agents, and the addition of thiols to the platelet homogenates increases the enzymatic activity. The reaction exhibits a pH optimum of 8.5-9.0. Divalent cations stimulate the reaction, and manganous chloride was the most effective of those investigated. The K(m) of the enzyme for myoinositol is 0.27 mM, and the K(m) for cytidine diphosphate diglyceride is 0.53 mM. The enzymatic activity of platelets isolated from patients with several diseases known to interfere with platelet clotpromoting function is similar to the enzymatic activity of platelets from normal donors.

The production of carbon monoxide from hemoglobin in vivo.

Dogs anesthetized with pentobarbital were shown to produce carbon monoxide at an average rate of 0.21 +/- (SD) 0.05 ml per hour. After intravenous injection of erythrocytes damaged by incubation with N-ethylmaleimide, CO was produced in excess of base-line production for 3 to 4 hours with an average yield of 0.89 +/- (SE) 0.046 mumole of carbon monoxide to 1 mumole of heme degraded. After intravenous injection of N-ethylmaleimide (NEM)-treated erythrocytes containing hemoglobin labeled with (14)carbon, (14)CO was produced. Its specific activity was approximately one-eighth that of the injected heme. It was also produced after intravenous injection of solutions of hemoglobin-(14)C and of reconstituted methemoglobin containing hemin-(14)C, but not after injections of methemoglobin containing globin-(14)C. The average yields of (14)CO from metabolized heme in the experiments with damaged erythrocytes and hemoglobin solutions were 89 +/- (SE) 4.6 and 97 +/- (SE) 17.0%, respectively. These results demonstrate that the CO produced during hemoglobin degradation arises from the heme moiety. The yield of (14)CO after injection of hemoglobin-(14)C solutions decreased significantly to values of 35 and 42% in two experiments when exogenous CO was added to the body stores, resulting in blood carboxyhemoglobin levels of 11.3 and 13.2% saturation. This finding suggests that oxidative metabolism is required during catabolism of hemoglobin to CO and that carboxy-hemoglobin levels in this range are sufficient to cause inhibition. After intravenous injection of either hemin-(14)C or protoporphyrin-(14)C, (14)CO was also produced. After injection of protoporphyrin-(14)C labeled bilirubin was isolated from gall bladder bile, and labeled hemin was isolated from the liver. It is thus very likely that protoporphyrin is converted to heme before the formation of CO. There was a large difference between the maximal rates of catabolism of hemoglobin to CO observed after injection of damaged erythrocytes and hemoglobin solutions. The limiting parameters in these processes are not yet clear.

Carbon monoxide production associated with ineffective erythropoiesis.

The rate of endogenous carbon monoxide production ( Vco), determined by the closed rebreathing system technique, was elevated above the normal range in four of five patients studied with ineffective erythropoiesis (four patients with primary refractory anemia, one with thalassemia). The mean molar ratio of Vco to Vheme (rate of circulating heme catabolism, determined from (51)Cr red cell survival curves) was 3.0 +/- 0.6 (SE), indicating that most of the CO originated from sources other than circulating erythrocyte hemoglobin, in contrast to previous findings in patients with hemolytic anemia, where Vco paralleled Vheme closely.After administration of glycine-2-(14)C to these patients, endogenous CO was isolated by washout of body CO stores at high pO(2) or by reacting peripheral venous blood samples with ferricyanide. The CO was then oxidized to CO(2) by palladium chloride and trapped for counting in a liquid scintillation spectrometer. "Early labeled" peaks of (14)CO were demonstrated which paralleled "early labeled" peaks of stercobilin and preceded maximal labeling of circulating heme. Production of "early labeled" (14)CO in patients with ineffective erythropoiesis was greatly increased, up to 14 times that found in a normal subject. The increased Vco and "early (14)CO" production shown by these patients are presumably related mainly to heme catabolism in the marrow. The possibility exists that hepatic heme and porphyrin compounds may also contribute significantly to Vco, as suggested by the finding of a high Vco in an additional patient with porphyria cutanea tarda.

Molecular characterization of two novel transporters from human and mouse kidney and from LLC-PK1 cells reveals a novel conserved family that is homologous to bacterial and Aspergillus nucleobase transporters.

Nucleobase transport is important for the metabolism of nucleic acids and antiviral and antineoplastic drugs. This transport has been functionally described in several mammalian cells but has not been well characterized molecularly. We report the cloning of two novel transporters. YSPL2 encodes a 650-residue protein and has an ubiquitous 8 kb transcript. The human and pig homologs are 95% similar. YSPL3 encodes a 598-residue protein with a 3 kb transcript that is expressed only in kidney and liver. Human YSPL2 and YSPL3 are 60% similar at the amino acid level and both show 31% similarity to the first nucleobase permease gene described in vertebrates, YSPL1. These proteins appear to be members of a new family of possible nucleobase transporters with significant sequence similarities with bacterial and Aspergillus nucleobase transporters. Further functional studies will be needed to unveil the role of these transporters in nucleic acid metabolism in normal and in disease states.

Purification of human peripheral blood colony forming cells (CFUC).

A 10-fold enrichment of colony forming cells (CFUC) from single donor platelet-apheresis residues and from 70-120 ml of peripheral blood of normal donors was achieved by sequential sedimentation on Ficoll-diatrizoate, depletion of cells adherent to plastic, and depletion of cells rosetting with sheep red blood cells (T lymphocytes). Culture of 5 x 10(5) unfractionated mononuclear cells yielded 9 +/- 3 colonies and mononuclear cells depleted of adherent cells and T lymphocytes yielded 53 +/- 6 colonies. The mononuclear cell fraction depleted of adherent cells and T lymphocytes was further enriched for CFUC by isopycnic sedimentation of Percoll gradients. Cells recovered in the 1.0063-1.065 g/cm3 density layer of the gradient formed 146 +/- 9 colonies in culture. The mononuclear cells depleted of adherent cells and T lymphocytes were also enriched for CFUC by depletion of Fc-receptor positive cells using an immune sheep red blood cell rosette sedimentation technique. Cultures of the Fc-receptor depleted fractions yielded 107 +/- 12 colonies, while the Fc-receptor enriched fraction yielded only 2 +/- 1 colonies. CFUC appear to lack surface membrane receptors for sheep erythrocytes and the Fc portion of immunoglobulin as well as the ability to adhere to plastic.

Rapid in situ cytochemical classification of CFU-C colonies in soft agar culture: permanent whole culture preparations.

Soft agar culture CFU-C (colony-forming units in culture) are rapidly classified in situ as eosinophil, macrophage, and neutrophil-monocyte types by whole culture staining with luxol fast blue for eosinophil specific granules and acetoorcein for nuclei. Stained colonies may be picked and examined individually as wet mount cover slip preparations, or the agar culture may be air dried and mounted permanently. The whole culture stain has been variously modified for the enzyme markers alpha-naphthyl acetate esterase and peroxidase and for nuclear staining alone with acetoorcein.

CD4 lymphocyte decline and survival in human immunodeficiency virus infection. The Military Medical Consortium for Applied Retroviral Research.

The loss of the CD4 lymphocyte is the central pathophysiologic event in the progression of human immunodeficiency virus (HIV) infection. This retrospective study, based on review of data from deceased HIV patients followed in a single HIV clinic, was conducted to determine if the rate of CD4 lymphocyte decline was predictive of survival. Forty of 172 patients met defined criteria for inclusion in this study. For each patient, CD4-cell counts showed approximate exponential decline over time. A Cox regression analysis was used to assess the association of CD4 cell decline (half-life), race, age, gender, initial CD4-cell count, and treatment (anti-Pneumocystis carinii pneumonia prophylaxis and/or zidovudine vs. no therapy) on total survival (from initial CD4 cell count) and on remaining survival time after reaching a CD4 cell count of 100 (estimated). For all patients, the rate of CD4 cell decline was predictive of total survival (p = .009) but not for survival after reaching a count of 100 (p = .6). For patients who had never received therapy (6 patients), however, the CD4 half-life remained associated with survival time from 100 CD4 cells (p < .05) as opposed to the treated patients. Therapy was the single variable most predictive of both survival endpoints, resulting in an increase in median total survival of 27.2 mo (p < .00001) and of 15.4 mo from a CD4 cell count of 100 (p < .00004). Nonwhites had a slight survival disadvantage compared to whites (p = .08 overall; p = .02 from CD4 cell count of 100).(ABSTRACT TRUNCATED AT 250 WORDS)

United Kingdom Beryllium Registry: mortality and autopsy study.

This report is based on 30 deaths from chronic beryllium disease (CBD) in the United Kingdom with details of 19 autopsies. The majority were fluorescent lamp workers and machinists who died from respiratory failure. There were no cases of lung cancer. The survival times ranged from less than 1 to 29 years and was longest in machinists. All of the workers showed interstitial pulmonary fibrosis with varying degrees of cystic change. The majority showed hyalinized, and a few active sarcoid-type, granulomas. Extrathoracic granulomas, as in a U.K. sarcoid autopsy series, were rare. A notable difference was the absence of myocardial involvement in CBD compared to an incidence of 20% in the sarcoid autopsies. The detection of beryllium in the criteria for diagnosis is emphasized and the cases classified as definite include 12 of 19 positive analysis, 6 of 19, negative or unavailable analysis. The remaining case was classified as dubious because, despite a positive analysis, granulomas were absent. The main differential diagnosis is sarcoidosis.

New aid for diagnosing chronic beryllium disease (CBD): laser ion mass analysis (LIMA).

The value of laser ion mass analysis (LIMA) for the detection of beryllium disease in routine histological sections was investigated. Our results were based on the examination of 14 cases of confirmed and five cases of suspected chronic beryllium disease. With the exception of two cases of coal workers' lungs, all the control material, normal, tuberculous, and sarcoid lungs examined were free of beryllium. The technique may also be useful in other diseases of possible occupational origin.

"Residual bodies" in sarcoid and sarcoid-like granulomas.

A MORPHOLOGICAL AND HISTOCHEMICAL STUDY WAS MADE OF EPITHELIOID CELL GRANULOMAS: (a) classical sarcoid type, namely, sarcoidosis, Kveim tests, tuberculosis, farmer's lung, and Crohn's syndrome; (b) sarcoid-like granulomas, often distinguishable from (a) by the presence of extracellular mucin or bile, namely, ulcerative colitis, diverticulitis, cholecystitis, cholangitis, carcinoma of the rectum and lymph nodes, draining tumours. All these granulomas showed similar, numerous cytoplasmic granules in epithelioid and giant cells with the properties of residual bodies, i.e., end products of activated lysosomes. The presence of residual bodies demonstrates the following features the morphological similarity of the granulomas studied, and the phagocytic nature of the affected cells. It suggests a common mechanism of granuloma formation but does not identify any particular exogenous cause. The findings suggest that Boeck's sarcoidosis may be caused by unidentified exogenous agents.

Comparison of lymphocyte transformation and macrophage migration inhibition tests in the detection of beryllium hypersensitivity.

In seven patients with chronic beryllium disease (Be) the Be lymphocyte transformation test was positive in 100%, independent of steroid therapy, and was reproducible. The Be macrophage migration inhibition test was only positive in four of seven patients (57%) not on steroids, and was not reproducible. In 72 potentially exposed healthy beryllium workers the lymphocyte transformation test was negative in all subjects. The macrophage test was positive in four of 78 and again the results were not reproducible. The workers with positive results showed no differences in age, type or duration of employment from those with negative results and showed no evidence of disease. In addition, the macrophage test was positive in two of 45 non-exposed control subjects. We also confirmed the above advantages of the lymphocyte transformation technique by using tuberculin antigen (PPD). The PPD lymphocyte transformation test gave positive results in approximately 60% of healthy beryllium workers, but the PPD macrophage test was only positive in 7%. We conclude that the Be lymphocyte transformation test is the most sensitive and reproducible and advocate its use in the diagnosis of disease and in monitoring the health of potentially exposed workers.

Granulomatous inflammation--a review.

The granulomatous inflammatory response is a special type of chronic inflammation characterised by often focal collections of macrophages, epithelioid cells and multinucleated giant cells. In this review the characteristics of these cells of the mononuclear phagocyte series are considered, with particular reference to the properties of epithelioid cells and the formation of multinucleated giant cells. The initiation and development of granulomatous inflammation is discussed, stressing the importance of persistence of the inciting agent and the complex role of the immune system, not only in the perpetuation of the granulomatous response but also in the development of necrosis and fibrosis.

The Kmif (Kveim-induced macrophage migration inhibition factor) test in sarcoidosis.

Circulating lymphocytes from 30 patients with sarcoidosis when stimulated in vitro with Kveim-induced macrophage migration factor, the Kmif test, produced a guinea-pig macrophage migration inhibition factor in 21 of 30 cases (70%). In those patients not on steroids the results showed a good correlation with the cutaneous Kveim test. One positive test was found in 16 normal subjects. Our results suggest that the Kmif test may prove a useful rapid alternative to the Kveim test.

Role of in vitro and in vivo tests of hypersensitivity in beryllium workers.

The value of the beryllium macrophage migration inhibition (Be MIF) and Mantoux tests in the diagnosis of chronic beryllium disease and in the detection of hypersensitivity in healthy beryllium workers is demonstrated. In the absence of steroid treatment the Be MIF test is positive in chronic beryllium disease patients. Seven of 50 (14%) helathy beryllium workers were Be MIF positive, while all the control subjects, normal and sarcoidosis patients, were negative. Healthy beryllium workers tend to be more often Mantoux negative than a comparable group of non-exposed workers, and although not conclusive this finding correlates with a positive Be MIF test. Although the detection of hypersensitivity is not diagnostic of disease, the Be MIF test can be used as an additional method for monitoring the health of beryllium workers. The full significance of our results should be assessed by a long-term study.

Halothane macrophage migration inhibtiion factor test in halothane-associated hepatitis.

As an index of delayed hypersensitivity in vitro halothane macrophage migration inhibition factor tests (halothane-MIF tests) were performed on peripheral blood lymphocytes from five patients with halothane hepatitis. Twenty-two subjects exposed to halothane, but with no evidence of jaundice, five 'healthy' hospital anaesthetists, nine jaundiced subjects without halothane exposure, and 10 healthy subjects with no history of exposure to halothane were also tested. The halothane-MIF test was positive in four of the five patients with halothane-induced hepatitis; the negative result was in a patient on steroid treatment. The test was negative in all other subjects. Our findings suggest that the halothane-MIF test may be of value in the diagnosis of halothane-induced hepatitis and as a screeening procedure for the identification of susceptible subjects.

The first seven cases of chronic beryllium disease in ceramic factory workers in Japan.

In the present paper the first 7 cases in Japan of chronic beryllium disease found in workers employed in a ceramic factory utilizing beryllium have been described. Immunological examinations of these cases showed changes similar to those observed in sarcoidosis, that is, negative tuberculin test and increase in serum gamma globulin and immunoglobulins. The fact that a considerable number of workers in the same factories as the patients showed negative tuberculin reaction may suggest that there may be further cases of chronic beryllium disease among them that are still in a latent period.

International Kveim histology trial.

An histological analysis of Kveim-test biopsies performed on a total of 306 patients including those with sarcoidosis, Crohn's disease, tuberculosis, and collagen disease is presented. Five main Kveim suspensions, "antigens," were used in the survey, Colindale K 12, Lot 5, Marburg 2 and 3, Commonwealth Serum Laboratory (CSL) early batches, 001/2/3 and late batches, 005/6. The biopsies were read blind by two observers (overall agreement 75%), the nature of the antigen and disease later disclosed, and the finding statistically analysed. It was agreed that focal epithelioid-cell collections remain as the essential criteria for the diagnosis of Kveim positivity. Diffuse epithelioid cells, Langhans' giant cells, lymphocytes, and necrosis are associated with focal epithelioid cells and hence Kveim positivity. Steroids were found to inhibit Kveim positivity. Observer disagreements were mainly due to difficulties in distinguishing macrophages from true epithelioid cells. We conclude that doubtful cells should be ignored in deciding Kveim positivity. Lymph-node-derived Kveim antigens were the most sarcoid-specific. They were more granulogenic and produced more necrosis, despite also producing the most foreign material. It is recommended that validation of new Kveim antigens must include adequate testing on patients with Crohn's disease, as the more nonspecific antigens were shown to give equal positive rates in sarcoidosis and Crohn's disease.