Cytoplasmic type 80S ribosomes associated with yeast mitochondria. IV. Attachment of ribosomes to the outer membrane of isolated mitochondria. Growing yeast spheroplasts were shown to have, on the average, four times the number of cytoplasmic ribosomes in contact with the outer mitochondrial membrane compared to starved spheroplasts. Ribosomes in contact with mitochondria in the growing spheroplast preparation, like free cytoplasmic ribosomes, exist primarily as polysome structures. In the starved spheroplast preparation, both mitochondria-bound and free cytoplasmic ribosomes exist primarily as monosomes. Mitochondria isolated from growing spheroplasts in a medium containing lmM Mg++ have cytoplasmic ribosomes bound directly to the outer membrane. These ribosomes can be quantitatively removed by washing the mitochondria with 2 mM EDTA. Mitochondria from starved spheroplasts are capable of accepting either free cytoplasmic polysomes or cytoplasmic polysomes extracted from mitochondria. However, the extent of polysome binding to mitochondria was shown to be a direct function of the Mg++ concentration; a smaller percentage of the input polysomes bind as the Mg++ concentration is lowered. At 1 mM Mg++, neither free cytoplasmic nor mitochondria-bound polysomes bind to mitochondria. Nevertheless, when growing spheroplasts are broken and mitochondria isolated in medium containing 1 mM Mg++, the mitochondria are seen to have cytoplasmic ribosomes firmly attached to the outer membrane. This result, in addition to our earlier data (Kellems, R. E., and R. A. Butow. 1974. J. Biol. Chem. 249:3304-3310), support the view that cytoplasmic ribosomes attached to the outer membrane of purified mitochondria were attached in vivo. In preparations of mitochondria isolated from growing spheroplasts, ribosomes appear to be found to specific regions of the outer membrane, namely those regions which are in close association or in contact with the inner mitochondrial membrane. This is particularly evident with mitochondria in a condensed configuration. This finding suggests a mechanism whereby cytoplasmically synthesized mitochondrial protein could be transferred by a process of vectorial translation across both membranes of the organelle.


Simplified technique for the insertion of indwelling catheters in the pulmonary artery and left atrium. A technique for the insertion of Telfon catheters into the pulmonary artery and left atrium is described. The method is especially useful for continous monitoring of pulmonary artery and left atrial pressures and sampling of mixed venous and arterial blood after open or closed heart surgery. No cpmplication associated with this technique have occurred in more than 100 clinical cases.


Concanavalin A receptors, immunoglobulins, and theta antigen of the lymphocyte surface. Interactions with concanavalin A and with Cytoplasmic structures. The effect of concanavalin A (Con A) on the capping of mouse lymphocyte surface immunoglobulin (surface Ig), cross-linked by rabbit anti-mouse Ig antibody, and on the capping of mouse thymocyte theta antigen, cross-linked by anti-theta alloantibody and rabbit anti-mouse Ig antibody, has been studied by immunofluorescence, using fluorescein conjugated Con A and rhodamine-conjugated anti-mouse Ig antibody, and by electron microscopy, using native or fluorescein-conjugated Con A and ferritin-conjugated anti-mouse Ig antibody. Prior incubation of the cells with Con A inhibited only partially capping os surface Ig, whereas it blocked almost completely capping of theta antigens. Both on cells with rings and on cells with caps the staining for surface Ig or theta antigen was superimposed to the staining for Con A. When Con A receptors on spleen cells were capped by Con A at concentrations of 10 mug/ml or higher, and the distribution of surface Ig was examined under noncapping conditions, all detectable surface Ig were found in the caps. As shown by electron microscopy, surface Ig remained dispersed in a layer of Con A. The ability of Con A to cap surface Ig was not altered by the presence of cohchicine or vinblastine. These results suggest that surface Ig are cross-linked by Con A to other Con A receptors. In these conditions surface Ig behave essentially as Con A receptors, as for example, in their sensitivity to cytochalasin B during inhibition or reversal of capping induced by this drug. The behavior of surface Ig parallels that of Con A receptors also in the presence of vinblastine. It is concluded that in the presence of Con A, antimitotic drugs do not modify directly the interaction between Con A receptors and surface Ig, but probably influence the capping ability of the Con A receptors or, more in general, affect the ability to elicit movements over the cell surface. The role in capping of cytochalasin-sensitive and vinblastine-sensitive structures is discussed. Both types of structures appear to play an active role in the formation of a cap, although the former probably corresponds to the main mechanical system responsible for the active displacement of cytoplasmic and surface material.


Response of myogenic and fibrogenic cells to cytochalasin B and to colcemid. I. Light microscope observations. Cytochalasin B (CB) induces a biphasic retraction is some cell types. The rapid response that peaks in 30 min leads to the "dendritic" condition. Replicating myogenic and fibrogenic cells, as well as postmitotic myoblasts and myotubes, participate in this reaction. This is followed by a slower phase that requires 40 h for stabilization and leads to the fully "absorized" state. Only replicating myogenic and fibrogenic cells participate in this reaction. Postmitotic myoblasts and myotubes do not arborize but round up and float off into the medium. Pretreatment with Colcemid does not block the rapid response to CB, but does block arborization. CB-arborized cells exposed to Colcemid while in the presence of CB develop sufficient tension to pull themselves apart. If CB depolymerizes actin-like filaments, and if such filaments constitute the only contractile system in the cell, then it is difficult to visualize how cells in CB develop such tension. Colcemid induces twisting, birefringent bands in interphase- and metaphase-arrested myogenic and fibrogenic cells, and in postmitotic myotubes. Such bands are more evident when CB-arborized cells are removed from CB and allowed to relax in Colcemid. These birefringent bands assemble in the prescence of cycloheximide, and may constitute 20% of the volume of the cell.


Nucleolar necklaces in chick embryo fibroblast cells. II. Microscope observations of the effect of adenosine analogues on nucleolar necklace formation. The round nucleoli of chick embryo fibroblast cells, when exposed to adenosine (2 mM)or to a number of adenosine analogues, lose material and unravel over a period of several hours to become beaded strands, 20 mu M in length, termed nucleolar necklaces (NN). Light microscope observations on this process are described. Biochemical experiments have revealed that most of these analogues interfere with both messenger RNA synthesis and ribosome synthesis, causing extensive degradation of the preribosome species containing 32S RNA although most of the preribosomes containing 18S RNA survive. We suggest that it is the depletion from the nucleolus of the adhesive 32S and 28S RNA preribosomes which allows the remaining nucleolar apparatus to spread apart into the NN configuration. Also required for the maintenance of the NN structure is the synthesis of some ribosomal RNA (rRNA) possibly present as rRNA "feathers" on the DNA. The addition of inhibitors of rRNA synthesis such as actinomycin D to the NN-containing cells causes loss of rRNA. Then a contraction and collapse of the NN structure into small dense spheres is observed.


Studies of intercellular invasion in vitro using rabbit peritoneal neutrophil granulocytes (PMNS). I. Role of contact inhibition of locomotion. Intercellular invasion is the active migration of cells on one type into the interiors of tissues composed of cells of dissimilar cell types. Contact paralysis of locomotion is the cessation of forward extension of the pseudopods of a cell as a result of its collision with another cell. One hypothesis to account for intercellular invasion proposes that a necessary condition for a cell type to be invasive to a given host tissue is that it lack contact paralysis of locomotion during collision with cells of that host tissue. The hypothesis has been tested using rabbit peritoneal neutrophil granulocytes (PMNs) as the invasive cell type and chick embryo fibroblasts as the host tissue. In organ culture, PMNs rapidly invade aggregates of fibroblasts. The behavior of the pseudopods of PMNs during collision with fibroblasts was analyzed for contact paralysis by a study of time-lapse films of cells in mixed monolayer culture. In monolayer culture, PMNs show little sign of paralysis of the pseudopods upon collision with fibroblasts and thus conform in their behavior to that predicted by the hypothesis.


Electron-microscopic mapping of AT-rich regions and of E. coli RNA polymerase-binding sites on the circular kinetoplast DNA of Trypanosoma cruzi. Partial alkaline denaturation of the circular kinetoplast DNA (kDNA) of Trypanosoma cruzi has shown the existence of 4 small, well-defined AT-rich regions with an average size of about 200 base pairs. They are almost equally distributed, separated by approximately 90 degrees on the circular molecule. All minicircles, whether free or linked in networks, have the same denaturation pattern and, therefore, seem to contain the same information. The long linear molecules present in low amounts in the kDNA samples do not show the same denaturation pattern. Partial denaturation of molecules in larger associations indicates that the circular units may be linked to each other by one strand only. kDNA can be transcribed in vitro by the RNA polymerase of E. coli. RNA polymerase-kDNA complexes have been studied in the electron microscope. By spreading the DNA-protein complexes by adhesion to positively charged carbon films and dark-field observation, it was possible to show the existence of 4 specific binding sites of the E. coli RNA polymerase on the kDNA circles. Comparing the position of the polymerase-binding sites and the AT-rich melted zones, it is suggested that a correlation exists between the two. As had been shown in earlier work, the replication of circular kDNA can be blocked by treating the trypanosomes with the trypanocidal drug Berenil. The comparison of the relative position of the Berenil-blocked replication forks with the position of the 4 denaturation loops shows that the DNA replication is stopped at these AT-rich regions. Since there is evidence that Berenil binds preferentially to AT-rich DNA and seems to be involved in inhibition of DNA replication, the following hypothetical model can be proposed. The replication of the circular kDNA molecules is discontinuous and involves the synthesis of RNA primers; when Berenil is bound to the AT-rich regions, synthesis of new RNA primers is inhibited and replication is blocked at these points, leading to the accumulation of replicating intermediates with defined branch lengths.


[Splenic emergencies and pregnancy].

[Splenic emergencies and pregnancy]. The authors report a personal case of spontaneous rupture of the splenic vein during pregnancy, already published elsewhere [6], and study again spontaneous rupture of the splenic vessels, artery or vein, and rupture of the spleen in relation to pregnancy, either during or after labour.  In all, 150 cases were found in the world literature.  This is an extremely dangerous surgical emergency, and the mother's life is in danger immediately.  We attempted to determine the clinical and paraclinical factors which would permit one to lay down the early indications for operation in these two emergencies.  Splenectomy remains, in all cases, the basis of surgical treatment.  In cases of maternal survival, the late, general and obstetric prognosis remains good.


Inhibitory action of somatostatin on pancreatic alpha and beta cell function. In normal men, the administration of somatostatin almost completely abolished the glucose stimulated insulin-release seen during control studies (without somatostatin), and caused a further reduction in glucagon secretion beyond that induced by hyperglycemia. Following the infusion, there was a rapid and marked rebound for insulin but not glucagon secretion. These events were attended by a marked retardation of the glucose disappearance rate (K) which exhibited two clearly separable components; the initial slow component (mean K equals 0.64) coincided with the period of insulin suppression and was followed by a faster component (mean K equals 1.37) temporally related to the marked rebound increase in insulin release after discontinuation of the somatostatin infusion. Similarly, the addition of somatostatin infusion completely blocked the release of insulin and growth hormone and delayed the release of glucagon stimulated by arginine infusion. Following the somatostatin infusion there was a small rise in GH and a marked rebound for insulin and this was associated with a higher level of plasma glucose than that found following arginine infusion alone. These data establish that the administration of somatostatin can effectively block the release of insulin stimulated by arginine and glucose, can attenuate the release of glucagon induced by arginine and can enhance the glucose-mediated glucagon suppression. The attendance of a relative hyperglycemia during these events is probably the net result of an impediment in peripheral glucose disposition due to acute insulin lack and a decreased hepatic glucose output secondary to glucagon suppression.


The significance of weight loss in the evaluation of pituitary response to LH-RH in women with secondary amenorrhea. Sixteen women with amenorrhea occurring in the setting of severe self-imposed weight loss and 18 women with secondary amenorrhea due to other causes were given LH-RH (luteinizing hormone-releasing hormone). Women with weight loss were found to be unresponsive to LH-RH when severely underweight. FSH responsiveness returned in a linear fashion as weight gain occurred and was not related to estrogen levels. LH responsiveness also returned with weight gain although the relationship was not linear but exponential and a sudden increase in responsiveness occurred at 15% below ideal weight. No relationship to estrogen levels could be found. Women who experienced amenorrhea in a setting other than weight loss did not demonstrate responsiveness to LH-RH which could be correlated with body mass, even when underweight. Women who experienced amenorrhea with weight loss had a consistently lower LH response to LH-RH than the second group and their LH response was always lower than the FSH response. On the other hand, a variety of patterns was found in women with amenorrhea due to other causes.


Inhibition of luteinizing hormone secretion induced by synthetic LRH by long-term treatment with glucocorticoids in human subjects. The response of luteinizing hormone to a single injection of synthetic LRH was established in 6 women and 7 men following an intramuscular dose of 0.2mg or 0.1mg. Significant increases  of serum LH were observed in 30 min. Pretreatment with metyrapone for one day (4.5 g/sugject injection of synthetic LRH (0.1 MG/SUBJECT). Seven women, 21-48 years of age who were treated with prednisolone for a least 11/2 months were examined for responsiveness of the anterior pituitary to a single injection of synthetic LRH (0.2 MG). The secretion of LH was markedly suppressed and maximal serum levels of LH WERE OBSERVED 90 MIN FOLLOWING LRH injection.


Isolated deficiency of follicle-stimulating hormone: Further studies. We record further studies over the past 2 yr on a unique female subject with isolated follicle-stimulating hormone (hFSH) deficiency, who developed human anti-gFSH antibodies after treatment with exogenous urinary gonadotropins. Administration of LRH resulted in a significant rise in serum hLH, but hFSH levels remained undetectable, "alpha Subunit" (the common alpha chain of the glycoprotein hormones) was detectable in basal samples obtained from our patient, and rose sharply after LRH. This is concordant with the hypothesis that the defect in our subject may be in the synthesis of the beta chain of hFSH, but it does not exclude other possibilities. The concentration of hFSH antibodies in the patient's serum has been monitored and her response to a further course of exogenous gonadotropins is recorded. The anti serum exhibits specificity for the hFSH molecule; the alpha and the beta chains of hFSH are virtually inert in competing with tracer 125-I-hFSH for binding to the antibody.


The diagnosis of Hashimoto's thyroiditis. Computer methods were used to estimate the usefulness of several clinical signs and laboratory tests in the diagnosis of chronic lymphocytic (Hashimoto's) thyroiditis. Information was drawn from the records of 217 patients with this disease seen at two hospitals in Southern California. The parameters studied included the physical characteristics of the thyroid gland; the appearance of the radioisotope thyroid scan; the response to a perchlorate discharge test; the serum antithyroglobulin antibody titer; the serum TSH concentration measured by radioimmunoassay; the thyroid radioiodine uptake response to exogenous TSH stimulation; and the serum PBI-T4I difference. Of the above, the PBI-T4I difference was deleted, being of limited value. The TSH stimulation test and serum TSH measurement were considered as alternative ways to evaluate thyroid reserve. Therefore, five diagnostic markers remained, all useful but not definitive. Computer and rule-of-thumb methods (two or more of the five markers positive) were tested for successful diagnosis of 145 patients with Hashimoto's thyroiditis proven by pathological examination of biopsied tissue; 23 of these were patients not used in the original pool of data. As a rule of thumb, it was found that Hashimoto's thyroiditis is the likely diagnosis if two or more out of the five useful markers are in its favor. For best results, four and preferably all of the five criteria should be tested. If this is done, the expected diagnostic accuracy for patients with Hashimoto's disease is by the rule-of-thumb method 67% correct, 21% indecisive, and 12% false negative; by computer methods it is 88% correct, 4% indecisive, and 8% false negative. By each method the number of false positive diagnoses was equivalent to 25% of the total number of patients with this disease. The false positive results nearly all occurred in patients with goiter associated with a defect in thyroid hormone synthesis.


Effect of synthetic luteinizing hormone releasing hormone (LH-RH) on the release of gonadotropins in Cushing's disease. The effect of intravenous synthetic luteinizing hormone releasing hormone (LH-RH) on plasma radioimmunoassayable levels of gonadotropins was investigated in 6 women suffering from Cushing's disease with bilateral adrenal hyperplasia. In five of six cases no significant variation of plasma LH levels was found following stimulation; in one case the response to LH-RH was present although slightly reduced below the normal range. By contrast in all cases the plasma FSH response was similar to that recorded in normal subjects. The explanation of impaired LH response is not clear but the possibility that endogenous hypercortisolism affects the pituitary responsiveness to LH-RH has to be considered.


Abnormal pancreatic alpha-cell function in first-degree relatives of known diabetics. A standard intravenous glucose tolerance test utilizing a 25g glucose load was administered to 15 first-degree relatives of known diabetics and 19 normal control subjects with no known diabetic relatives. Despite normal glucose tolerance, 5 of the first-degree relatives had a significantly lower release of first-phase insulin (p 0.05). In contrast to 8 normal subjects and the remaining 10 first-degree relatives the secretion of glucagon by the 5 first degree relatives with abnormally low insulin release was not suppressed by induced hyperglycaemia. These findings provide evidence of abnormal alpha-cell function in a very early stage of genetic diabetes.


Studies with human leukocyte lysosomes. Evidence for antilysosome antibodies in lupus erythematosus and for the presence of lysosomal antigen in inflammatory diseases. Human lysosomes were isolated from normal peripheral blood leukoyctes and characterized by electron microscopy, enzyme analysis, and assays for DNA and RNA. Stored sera from 37 unselected patients with systemic lupus erythematosus (SLE), including active and inactive, treated and untreated cases, were tested in complement fixation (CF) reactions with these lysosome preparations. 23 SLE sera exhibited positive CR reactions, as did sera from two patients with "lupoid" hepatitis. The seven SLE sera with strongest CF reactivity also demonstrated gel precipitin reactions with lysosomes. Neither CF nor precipitin reactions with lysosomes were observed with normal sera or with sera of patients with drug-induced lupus syndrome, rheumatoid arthritis (RA), polymyositis, or autoimmune hemolytic anemia. By several criteria the antilysosome CF and precipitin reactions of SLE sera cound not be attributed to antibody to DNA, RNA, or other intracellular organelles. The lysosomal component reactive with SLE sera in CF assays was sedimentable at high speed and is presumably membrane associated. The CF activity of two representative SLE sera was associated with IgG globulins by Sephadex filtration. A search for lysosomal antigen in SLE and related disorders was also made. By employing rabbit antiserum to human lysosomes in immunodiffusion, a soluble lysosomal component, apparently distinct from the sedimentable (membrane-associated) antigen described above, was identified in serum, synovial fluid, or pleural fluid from patients with SLE, RA, ankylosing spondylitis, and leukemoid reaction. An antigenically identical soluble component reactive with the rabbit antiserum could be released in vitro from intact lysosomes by repeated freeze-thaw cycles..


Utilization of cultured human lymphoid cells for detection of humoral sensitization in prospective recipients of kidney transplants. Prospective recipients of kidney transplants were tested for lymphocytotoxicity; from these we selected 102 sera that lacked cytotoxic antibodies against peripheral lymphocytes from at least 80 unrelated subjects. To detect humoral sensitization, we then reacted these with 17 cultured human lymphoid cell lines having different HL-A phenotypes. Cytotoxic antibodies reacting with these cultured cells were now detected in some of the sera. These antibodies were not directed against HL-A antigens, yet mediated lysis of target cells in the presence of rabbit but not of human or guinea pig complement. Furthermore, they activated the classical pathway of the rabbit complement system. Later, a significant association was found between occurrence of cytotoxic antibodies and rejection of the transplant. Thus, cultured human lymphoid cells, because of their great susceptibility to complement-mediated lysis, appear to be useful in detecting humoral sensitization in candidates for kidney grafts.


Immunopathological changes in rheumatoid arthritis and other joint diseases. A comparative study of the distribution of immunoglobulins G, M, and A and C3 in the synovium and inside synovial fluid leucocytes and of the relative levels of IgG, IgM, AND C3 in paired samples of serum and synovial fluid from both seropositive and seronegative patients with rheumatoid arthritis and other types of non-infective synovitis shows that although there is no distinctive immunopathological feature of rheumatoid arthritis, the incidence of immune complexes containing IgG and IgM with and without detectable C3 in the affected synovium or inside synovial fluid granulocytes is higher in rheumatoid arthritis and especially so in seropositive cases. The mean level of C3 in synovial fluid from patients with rheumatoid arthritis is lower than that from the group without rheumatoid arthritis. In contrast to previous reports, extracellular clumps of IgA could be detected in the affected synovium of a number of affected patients. Aggretated human IgG could be bound by some of the synovial biopsies and synovial fluid leucocytes from both seropositive and seronegative rheumatoid arthritis patients. Antinuclear factor and rheumatoid factor could be detected in the synovial fluid but not in the serum of several patients suggesting either selective sequestration or local synthesis of antinuclear and rheumatoid factors in the affected joints.


The distribution of muramidase (lysozyme) in human tissues. The distribution of muramidase (lysozyme) in normal and pathological human tissues has been studied, using an immunohistological technique. The enzyme was demonstrated in a variety of healthy tissues, including serous salivary acinar cells, lactating mammary tissue, Paneth cells, renal tubular cells, myeloid cells (including eosinophils), and histiocytic cells. In pathological tissues the most striking positivity was encountered in reactive histiocytic cells in granulomatous conditions such as tuberculosis and Crohn's disease. The finding of this study are related to previous reports of the distribution of human and animal muramidase and the implications of patterns of muramidase staining in pathological histiocytes are briefly discussed.


Isolations of aerobic sporing bacilli from the tips of indwelling intravascular catheters. Following previous reports of the isolation of aerobic sporing bacilli from intravenous catheters and infusion sets, it was decided to analyse a series of catheter tip cultures to ascertain the significance of these isolations. Those catheters from which aerobic sporing bacilli were isolated behaved statistically differently from other catheters from which organisms were isolated, but were statistically similar to sterile catheters. Using the nitroblue tetrazolium (NBT) test no significant difference was found between patients with isolates of aerobic sporing bacilli and those with sterile catheters. The NBT test readily distinguished those patients from whom other organisms were isolated. In a small number of instances some evidence of a possible pathogenic role for the aerobic sporing bacillus was found, but the results clearly indicate that in most cases these organisms can be regarded as contaminants.


Observations on the accuracy of point counting including a description of a new graticule. The accuracy of point counting as a histometric technique depends very much on the number of points counted, and the number necessary varies greatly with both the level of accuracy required and the proportion the relevant tissue occupies in the section. It is shown that the number of "hits" is a more useful measure than the total number of points counted, and a simple rule derived for determining the number of hits necessary to achieve any necessary degree of accuracy. A method of progressive estimation of accuracy when attempting to measure the proportion of a tissue that is unevenly distributed is also described. The point-counting graticules commonly used, with 25 points, are inadequate with tissue proportions below 10%. A new graticule design, with 100 points optimally distributed, and usable as a 25-point array if required, is illustrated.


Immunofluorescence in duodenal mucosa of children with acute enteritis due to a new virus. Electron microscopy od duodenal mucosa from children with acute non-bacterial enteritis has shown virus particles in epithelial cells. Indirect immunofluorescent techniques applied to the same tissue showed virus antigen localized in the cytoplasm of epithelial cells of the villi. Specific IgM antibody was present in sera from infected patients as early as two days after the onset of symptoms. Virus particles from different patients appeared to share a common antigen. The evidence presented supports our belief that this new virus was the cause of acute enteritis in the children studied.


Cell receptor studies on seven cases of diffuse histiocytic malignant lymphoma (reticulum cell sarcoma). Expression of B and T lymphocyte receptors has been studied in seven cases of reticulum cell sarcoma. In one case, surface receptors and tests of phagocytic function demonstrated the histiocytic origin of the neoplastic cells. In four cases, tumour cells expressed both B and T lymphocyte markers (two cases) or showed a normal pattern of expression of B and T lymphocyte markers. In the other two cases, lymphocyte receptors were not detected, and there was no evidence of phagocytic function: this class of receptor-silent tumours is of uncertain pathogenesis. The significance of these observations is discussed.


Sisomicin: Bacteriological and clinical evaluation. A preliminary study was conducted with sisomicin, a new aminoglycoside antibiotic. The drug was administered to 40 patients in doses varying from 1.5 to 3.75 mg/kg/day. Sisomicin proved to be an effective therapy in urinary tract infections and to a lesser extent in wound infections caused by gram-negative rods; favorable results have been observed in 91.6% and in 66.6% of the patients presenting these infections. Toxic reactions involving the hearing function, renal function, and general tolerance were infrequent: they occurred in less than 5% of the patients in this series. (However in approximately 22% of the patients, there was a transient appearance of granular casts in the urine.


The bioavailability of meperidine using urine assays for meperidine and normeperidine. The results of a two-way crossover trial of two commercial preparations of meperidine hydrochloride in 24 healthy volunteers is reported. The urinary excretion patterns resulting from dosing with the two preparations were followed for 48 hours, and the urine concentrations of meperidine and a major metabolite, normeperidine, were determined by gas liquid chromatography. Statistical analysis of the data showed no significant difference between the urine concentrations of meperidine and normeperidine produced by either preparation, and the preparations are of comparable bioavailability. The use of the urinary excretion pattern and concentrations represents a rapid, accurate, and quantitative method for determining the bioavailability of different preparations of meperidine hydrochloride.


Loxapine in newly admitted chronic schizophrenic patients. The standard drug Stelazine (STEL), at a dose of 50 mg/day, exhibited therapeutic activity significantly different from placebo (PL) activity on several variables,  most notably BPRS, attesting to the sensitivity of the experiment. On the other hand, the investigational drug, loxapine (LOX), in doses of 100 mg/day for four weeks, could be differentiated from PL as treatment in the described population on only one variable (NGI-Imp.) and one item of the BPRS. On several variables, positive trends were noted, but the differences from PL did not attain the critical values necessary for statistical significance at P smaller than 0.05. One might speculate that the relatively short duration of treatment in this study might account for the difference between these disappointing results and the more gratifying results of a previous loxapine study in chronic long-term institutionalized schizophrenics with the same oral dose.


Naproxen: long-term study in rheumatoid arthritis and "placebo pulse". Naproxen is by now a relatively well-known antirheumatic drug, and many short-term studies have shown its efficacy and relatively good tolerance. We have observed 64 patients with definite or classical rheumatoid disease, 27 of whom have been followed for well over two years on daily doses of naproxen to ascertain the persistence of drug efficacy and safety. In this part of our study, patients were subjected to complete clinical and biochemical evaluations at two-monthly intervals. Naproxen was well tolerated, and the few side effects reported were transient and mild in nature. Sequential laboratory studies revealed no significant anomaly. Clinical evaluation showed no pattern suggestive of decreasing antirheumatic activity. A question frequently encountered in the treatment of certain diseases such as rheumatoid arthritis is whether long-term improvement is due to efficacious suppressive therapy or spontaneous abatement of disease activity. We devised a double-blind placebo pulse phase in which 19 of our 28 long-term patients participated in a study within a study lasting four weeks. They were divided into two groups. The first group took their usual dose of naproxen during the first two weeks and a corresponding number of placebo tablets in the next two weeks. The procedure was reversed in the other group. We conclude that naproxen remains efficacious.


Naproxen and aspirin in rheumatoid arthritis: a multicenter double-blind crossover comparison study. One hundred nineteen adults with active definite or classical rheumatoid arthritis were studied in a multicenter double-blind crossover study of naproxen (500 mg/day) and aspirin (3.6 Gm/day). Each drug was given in sequence for a six-week study period. Patients already receiving corticosteriod and/or gold therapy were maintained at constant dose throughout the study, but analgesics and other nonsteroidal antiinflammatory agents were discontinued at baseline. Objective and subjective evaluations by both investigator and patient were carried out at two-week intervals. No significant difference in global evaluation of efficacy or individual measures of efficacy was observed between aspirin and naproxen therapy, although physicians' global evaluation tended to favor naproxen. Sedimentation rate was lower on aspirin (naproxen 43.1 mm/hr; aspirin 38.7 mm/hr; P=0.02). Naproxen, 250 mg twice daily, was significantly better tolerated than aspirin, 900 mg four times daily. Mild, moderate, and severe side effects were less frequent with naproxen. The incidence of heartburn was significantly lower on naproxen, and significantly fewer patients terminated their six-week study period on naproxen than on aspirin. There were no significant deviations from baseline values in hematocrit, white cell or differential counts, or in tests of renal and hepatic function during the course of the study.


A double-blind comparison of naproxen with indomethacin in osteoarthrosis. Twenty-two patients with osteoarthrosis of one or both knee joints and 28 patients with osteoarthrosis of one or both hips completed a double-blind trial of 500 mg naproxen daily versus 100 mg indomethacin daily. All patients had been on other active medication up to the start of the trial. Identical trial designs were followed with both classes of patients, namely, a crossover pattern of four weeks on each drug with patients being assessed at -2, 0, 2, 4, 6, and 8 weeks. Assessments made included objective measurement of joint range, stair climbing and walking times, and subjective grading of pain present during normal activity, of which the patient kept a daily record. Patients were also questioned at each clinic visit regarding possible side effects. Study groups were comparable for both drugs. In the majority of subjective and objective parameters, there were significant improvements from baseline on both drugs of statistically comparable magnitude. Significantly fewer side effects were noted during the period on naproxen compared with those on indomethacin. There were no abnormalities discovered in hematologic or biochemical tests performed during the course of the trial.


Ankylosing spondylitis: open long-term and double-blind crossover studies with naproxen. Participation in open and double-blind crossover studies in rheumatoid arthritis confirmed that naproxen improved pain and stiffness. This observation suggested that naproxen might be effective in ankylosing spondylitis. The initial trial was open, but at six months, a double-blind crossover "placebo pulse" was superimposed on the open trial. Thirty-six patients entered the trial taking a daily dose of 500 mg naproxen. At the end of the first month, 35 assessed naproxen as being equally effective to, or better than, previous therapy. The first ten patients to complete six months on naproxen took part in a placebo pulse study comprising two consecutive four-week periods, in one of which each patient took 500 mg naproxen, in the other, identical placebo capsules, the order being randomized. Eight patients correctly identified the placebo capsules (P=0.02). During the 16 months of trial, six patients have withdrawn, two being in remission and four for lack of efficacy. The remaining 30 patients have completed six months and 22 have completed at least 12 months on naproxen. At the end of six months pain was less (P=0.02), morning stiffness had decreased (P less than 0.01), and immobility stiffness had improved (P less than 0.01). These patients are impressed by the improvement in pain and stiffness and have little disability. All continue full-time employment and have been able to increase their leisure activities without discomfort. No persistent side effects were observed, and naproxen appears to be a useful drug in the treatment of anklyosing spondylitis.


The treatment of acute gout with naproxen. The effectiveness of naproxen in the management of acute gouty arthritis was assessed in an open study of 20 patients. These patients were selected on the basis of their clinical presentation of characteristic acute arthritis associated in 19 with concomitant hyperuricemia. There were 17 men and three women varying in age from 35 to 89 years. The first 12 patients were treated with 600 mg naproxen initially, followed by 300 mg every 8 hours for the first 48 hours and then tapered or discontinued depending upon their clinical response. The last eight patients received a loading dose of 750 mg naproxen, followed by 250 mg every 8 hours for a duration of 72 hours before tapering the drug. The response of 15 of the 20 was either excellent or good, while the response was fair in three and poor in two. Poor responders had been failures in other regimens or were treated late in the course of their attack. The higher loading dose was associated with more rapid and satisfactory remission. No significant undesirable side effects were observed. On the basis of this study, naproxen was found effective in alleviating the inflammation of acute gout.


Naproxen: corticosteroid-sparing effect in rheumatoid arthritis. Ambulatory patients with a diagnosis of "classical" or "definite" active adult rheumatoid arthritis were selected. They had been stabilized for at least six months previously with oral administration of paramethasone acetate plus various analgesics and/or nonsteroidal anti-inflammatory agents. During an initial period of two to five weeks, the paramethasone dosage was adjusted and other medications replaced by placebo to find the minimum required to keep the patient at a level similar to the one at the beginning of the study. Subsequently, with the double-blind method, the effect of 400 mg naproxen daily was compared with placebo, making periodic and progressive reductions in the corticosteroid dosage, unknown to the patient, until a tolerable minimum was found. The average total observation time was 14 weeks. Results obtained in the first 28 patients showed an average reduction of 57 per cent in paramethasone dose for the group treated with naproxen, versus 21 per cent for the control group. One patient experienced euphoria which could possibly have been related to the test drug. It is concluded that naproxen possesses a significant corticosteroid-sparing effect with negligible toxicity.


An open trial of naproxen in rheumatoid arthritis patients with significant esophageal, gastric, and duodenal lesions. To confirm the reported lack of major gastrointestinal side effects of naproxen, we gave 58 patients with active rheumatoid arthritis and significant gastrointestinal disease therapeutic doses of naproxen while closely monitoring them for signs and symptoms of gastrointestinal dysfunction. All patients underwent upper gastrointestinal x-ray examinations at the start of the trail, and, when indicated, during the course of the study. Endoscopies were also performed when indicated. Forty patients had hiatus hernia and 35 had peptic ulcer (23 duodenal ulcer and 12 gastric ulcer). Twenty-six patients had a combination of hiatus hernia with either type of peptic ulcer. After 262 patient visits over a period of 52 weeks, 35 patients remained in the study, all having had more than six months of naproxen therapy in dosages ranging from 500 to 750 mg daily. In 33 of the 35, the response to naproxen had generally been good to excellent. Only seven patients dropped out of the trial because of complaints referable to side effects. There were no major related upper gastrointestinal side effects as monitored by continuing clinical evaluation, stool occult blood, comprehensive laboratory examination, and, where indicated, upper gastrointestinal x-ray studies. Approximately 70 per cent of the patients demonstrated efficacy on long-term naproxen therapy by subjective and objective parameters. Naproxen appears to be an efficacious and remarkably safe drug in the long-term therapy of rheumatoid arthritis, even in the presence of significant upper gastrointestinal symptomatology.


Color-specific interconnections of cones and horizontal cells in the retina of the goldfish. In Golgi preparations of goldfish retina we have observed three types of horizontal cell which receive exclusively from cones and one which receives exclusively from rods. The cone horizontal cells were designated H1, H2 and H3, in order of increasing dendritic spread, increasing separation from the outer synaptic layer, decreasing size of perikaryon, and decreasing density of cone contacts. Slender appendages with knobby terminal enlargements project  horizontal cells by alalyzing serial 1 mum sections with the light microscope. The probable inputs, in terms of visual pigments in the cones which contact them, are: H1, red+green+blue; H2, green+blue; H3, blue. Analysis of previously published work suggests (1) that H1 cells generate monophasic or L-type responses, H2 cells generate biphasic or C1-type responses, and H3 cells generate triphasic or C2-type responses; (2) that H1 cells receive direct functional input at least from red-sensitive cones, H2 cells from green-sensitive cones, and H3 cells from blue-sensitive cones, and (3) that H1 constitute pathways from cones to H2 cells, and H2 cells, and H2 cells constitute pathways from cones and H1 cells to H3 cells. The precise location and route of the transfers, from H1 to H2 and from H2 to H3, are not yet known.


Horizontal cell axons and axon terminals in goldfish retina. Retinas of ordinary and black moor varieties of goldfish (Carassius auratus) were prepared by the Golgi method, mounted flat or sectioned vertically, and studied in the light microscope. Three types of horizontal cells whose dendrites contact only cones, and one type whose dendrites contact only rods, were observed. The cone horizontal cells (Cajal's "external horizontal cells") all have slender axons which descend gradually to the inner nuclear layer and terminate there in long, fusiform expansions (Cajal's "internal horizontal cells"). The thin and thick portions of the axons, as well as the perikarya of the horizontal cells, bear small numbers of straight, horizontally-directed, knobby filamentous appendages which may be sites of synaptic contact. The cone horizontal cell axons in goldfish, unlike those in higher vertebrates, do not terminate in contact with synaptic endings of photoreceptor cells, but in proximity to cells and processes deep in the inner nuclear layer. Axons have not yet been demonstrated on rod horizontal cells in goldfish.


The basilar pontine gray of the opossum: a correlated light and electron microscopic analysis. Neurons within the basilar pontine gray (BPG) of the American opossum can be subdivided into four major nuclei which are named medial, lateral, ventral and peripeduncular in accordance with previous studies. In addition, several smaller subnuclei, such as the median and dorsolateral cell groups, are present, as well as two longitudinal columns of neurons within the ventral nucleus. Neurons in the BPG range in size from 9 to 35 mu and appear randomly distributed so that none of the subdivisions contains exclusively nerve cells of the same perikaryal dimension. Projection neurons as shown in Golgi impregnations have a variable dendritic pattern; those in peripeduncular zones exhibit dendrites closely applied to the surface of the cerebral peduncle, whereas those in other regions generally have a radial type of arrangement. Certain projection neurons can be distinguished on the basis of their dendritic surface, which bears either claw-like protrusions or stalked appendages. Smaller nerve cells measuring less than 18 mu may be intrinsic neurons, since axon-like processes arise from their dendrites and course for some distance near the parent cell before becoming thin and beaded. Ultrastructural observations show profiles of neurons comparable in size to those seen in Golgi impregnations and suggest at least four classes of presynaptic profiles. One category ranges in size from 2 to 8 mu, contains round vesicles (average diameter 450 A) and characteristically forms multiple asymmetric synaptic contacts with several small postsynaptic profiles, some of which appear to be the dendritic claws mentioned above. The other three types of axon terminals measure less than 2 mu in their greatest dimension and are distinguished by their synaptic vesicles; one group containing round vesicles with an average diameter of 380 A, a second group exhibiting larger round vesicles with an average diameter of 500 A and a third group containing flatened or eliptical vesicles. Transection of the superior cerebellar peduncle produces early filamentous and later electron dense degenerative changes in some, but not all, of the larger types of presynaptic profiles. Subsequent to large motor-sensory cortex ablations both filamentous and dark degenerating profiles are simultaneously observed at all survival times. In one case with a cortical lesion restricted to the motor-sensory cortex, mainly dark degenerating terminals are apparent in the ipsilateral pontine gray, whereas in a lesion confined to the visual cortex only filamentous degeneration was observed. It is suggested, therefore, that some of the dark degenerating profiles represent the terminals of collaterals of corticospinal axons and the filamentous boutons are terminal expansions of direct corticopontine fibers.


The pattern of ocular dominance columns in macaque visual cortex revealed by a reduced silver stain. A pattern of alternating dark and pale bands was observed in the striate cortex of the macaque monkey. The bands, which ran parallel to the surface, were seen in tangential sections stained with a reduced silver method for normal fibers and were most clear in layer 4C alpha, immediately deep to the line of Gennari. The dark bands were about 300 mu wide and showed blind endings and bifurcations. The light bands were about 50 mu wide and did not branch or terminate within area 17. Because the dark bands were similar in width to the bands of terminal degeneration which have been shown to result from single-layer lesions of the lateral geniculate body, it seemed possible that they corresponded to ocular dominance columns. To test this idea, the boundaries of ocular dominance columns were marked in a physiological experiment: tangential electrode penetrations were made in an anesthetized monkey and, as the electrode was advanced horizontally in the fourth layer, the eye preference of single units and of the background activity was monitored. Small electrolytic lesions were placed at the points where a change in eye preference occurred. The brain was subsequently fixed, sectioned tangentially and stained with the silver method. All the lesions--total of 12 --fell directly on the pale bands. Moreover, the electrode had not passed over any pale bands without a lesion being placed. It was concluded that the dark bands do correspond to single ocular dominance columns and the pale bands to the boundaries between columns. The banding appearance is due to a greater density of tangential fibers within columns than at the borders of columns. These tangential fibers are in part the preterminal arborizations of geniculocortical axons, since some of them have been shown to degenerate after geniculate lesions. The ocular dominance columns were mapped for most of the striate cortex, using serial tangential sections stained with the silver method. The overall pattern was ,imilar in several monkeys, though the details of the branching arrangements varied from animal to animal. The columns met the 17-18 border at right angles. On the outer surface of the hemisphere the columns converged from the 17-18 border, turned medially with repeated fusions of columns, and streamed over the lip of the calcarine fissure. In the roof of the fissure they met a second system of columns oriented parasagittally. In terms of the visual field, the columns ran roughly horizontally for the central 10 degrees of the field, and circumferentially beyond that. The columns were not mapped in the stem of the fissure, the area corresponding to the far periphery of the field. The constancy of column width across the cortex probably allows a functional matching between ocular-dominance and orientation columns.


Structure and stability of casein micelles. The structure and stability of casein micelles are determined in large measure by the amino acid sequences of the constituent alphas1-, beta-, and kappa- caseins. In this paper we review, and attempt to connect with sequence data where possible: (1) molecular weight, structure, and dissociation of casein micelles; (2) physical characteristics of monomeric caseins, especially the distrubition of charged and hydrophobic residues and the possible occurrence of helical regions; and (3) forces involved in association of monomers.


A patient survey of denture tolerance before and after a mandibular vestibuloplasty with skin grafting. Thirty-six patients were examined by two prosthodontistis after a mandibular vestibuloplasty with skin grafting to determine whether there was a significant improvement in these patients ability to tolerate their dentures. Previous published reports have been made only by oral surgeons. The results indicate that this procedure is a reliable treatment modality for patients who complain of looseness, poor masticatory function, and pain.


Hospital dental service: a clinical evaluation to determine its justification. A research study was undertaken to determine the importance of a hospital dental service in civilian hospitals. Two hundred hospitalized patients were evaluated to determine their oral health status and possible effects of poor oral health on related physical situations. Both cental caries and periodontal disease were prelavlent in the hospital patient population, and 83.5% of the patients suffered from some type of oral pathosis that required treatment. Hospital dental care is important, not only for the health and improvement of the total patient but also for the hospital to provide complete patient care.


Management of gallstones in the aged.

Management of gallstones in the aged. Calculous gallbladder disease is the most common indication for intra-abdominal surgery in the elderly.  Feeding bile salts may possibly increase the solubility of cholesterol in bile and may cause diminution in the size of radiolucent gallstones, the principal ingredient of which is cholesterol.  However, indications for such treatment are still not clear, and long-term effectiveness and safety still have to be evaluated.  If the aged patient with gallstones has severe symptoms, is a good medical risk and is not demented, surgical treatment should not be denied because of advanced age.  If the patient has "silent" or "dyspeptic" gallstones and is a poor risk or demented, surgical therapy should be postponed at least temporarily, in the hope that medical methods for dissolving gallstones soon will be perfected.


Withdrawn, apathetic geriatric patients responsive to methylphenidate. A double-blind randomized study showed that methylphenidate, as compared to placebo, was singularly effective in 44 withdrawn, apathetic geriatric patients.  Study selection and exclusion criteria were rigidly enforced.  Patients receiving methylphenidate in a dosage of 20 mg daily improved significantly over a period of six weeks as measured by results of tests for mental status, ward behavior (nursis' rating), target-symptom response, and physician's and nurses' global evaluations.  For this reason and because there were no side effects, this form of antidepressant therapy should be considered in the treatment of previously unresponsive or oversedated patients.


Quantitation of isolated rat islets of Langerhans on the basis of deoxyribonucleic acid content under metabolic conditions of altered protein synthesis. A micromethod was employed to estimate quantitatively and reproducibly the deoxyribonucleic acid (DNA) content of isolated rat islets of Langerhans. The DNA content per islet varied linearly with the mean diameter or the dry weight of the islets isolated. The DNA in the freeze-dried islets of male Sprague-Dawley or Wistar rats was about 21.0 mug/mug islet dry weight. Three to four weeks after hypophysectomy, with or without short term bovine growth hormone replacement, the DNA content per unit dry weight of islets was not significantly altered. Islet DNA content and islet dry weight are proposed as an interconvertible and reliable basis of reference for measurements of different islet functions.


Study of rickettsioses in Slovakia. I. Coxiella burneti and Rickettsiae of the spotted fever (SF) group in ticks and serological surveys in animals and humans in certain selected localities in the Lucenec and V. Krtís districts. A study of the natural focus character of Q fever and the rickettsiosis caused by organisms of the SF group carried out in 1971-1973 in 7 selected villages in the Lucenec district and in a few control villages in the V. Krtis district, produced the following results: Among a total number of 2,200 ticks belonging to the species Dermacentor marginatus, Ixodes ricinus and Haemaphysalis punctata and tested for the presence of rickettsial organisms by the haemocyte test, C. burneti was detected in 8 from the chosen localities--in 2 females and 1 male of D. marginatus in Luborec, in 1 2. ricinus female in Maskova and in 1 female and 3 males of I. ricinus in Pincina--and in 5 from the control localities--in 1 D. marginatus female in Prebelce and in 3 females and 1 male of D. marginatus in V. Lom.


New concepts on tetanus immunization: naturally acquired immunity. Comments were made on misinterpretations concerning the natural resistance and natural immunization against tetanus. Arguments were given explaining why the disease itself did not determine adequate immunity. When, however, adequate conditions appear, tetanus toxin is known to stimulate the immune system and produce detectable humoral antibodies. Various possibilities resulting from the postulated harboring of tetanus bacilli by the human body and their eventual toxin production were analyzed and related to the human tetanus pathology. The existence of natural immunization was unquestionably demonstrated by presence of protective levels of tetanus antitoxin in the blood of the majority of 59 surveyed subjects considering that none of them had ever received any tetanus toxoid and most of them never received a single shot of any drug. The results of this survey originated a few arguments that may support the answer to some still intriguing phenomenona such as: 1. The relatively small number of cases of overt disease among people and animals born and living in large tetanus-risk regions all over the world. 2. The existence of "poor responders" and "good responders" to the primary tetanus toxoid stimulus. 3. The age distribution of tetanus showing evident prevalence among newborns and children. 4. The wide individual variations in the clinical picture of human tetanus as indicated by the localization and limitation of the symptoms and their severity.


Complex surveillance of Streptococcus pyogenes. I. Immunological surveys of anti-M antibodies and possibilities of long-term epidemiological prognosis. The results of complex surveillance of Streptococcus pyogenes, based on mass infections in the Czechoslovak army over a period of 11 years, are submitted. The epidemiological method revealed an incidence of cycles, lasting several years, of predominant pairs of T types, most of which could not be typed by their M protein. Auxiliary immunological surveys allowed the biological M activity of widespread strains to be estimated and probably form a basis for considerations on a negative long-term epidemiological type prognosis. The preliminary results presented offer prospects for a realistic methodological approach to the question of the type content of a future antistreptococcal vaccine.


Thymic involution: effect on T cell differentiation. The thymus of long-lived BC3F mice involutes progressively throughout life, beginning at 6 weeks of age. This is manifested by the loss of cortical lymphoid mass and by the degenerative changes in epithelial cells. The purpose of this study was to determine to what extent age-related degenerative changes of the thymus affect its capacity to influence the maturation of thymic-derived (T) cells. Accordingly, thymic lobes of mice ranging in age from 1 day to 33 months were implanted under the kidney capsule of T cell-deprived syngeneic young adult TXB mice, and the emergence of T cells was assessed kinetically by various morphologic and functional indices which may be reflective of different T cell subpopulations. They are: a) histology of the thymsu graft, b) lymphocyte repopulation of the T cell-dependent areas of lymph nodes, c) total number of splenic lymphocytes carrying theta antigens (theta-+), d) T cell-dependent humoral immune response and e) proliferative response of splenic cells to plant lectins, phytohemagglutinin (PHA) and succinyl-concanavalin A (s-Con A), and allogeneic lymphocytes. The results revealed that the T cell-transforming influence of thymic tissues generally decreases with increasing age. The difference in the patterns of recovery of the various indices of thymus-grafted TXB mice suggests that the extent to which T cells can mature is dependent upon the degree of involution the thymic tissue has undergone with age. In particular thymic tissues lose the capacity to influence the following functions with advancing age: 1) lymphocyte repopulation of the T cell-dependent areas of lymph nodes; 2) mitogenic reactivity of splenic cells to T cell-specific mitogens (PHA and s-Con A): 3) number of splenic theta-+ lymphocytes and splenic T cell helper function; and 4) mitogenic reactivity of splenic T cells to allogeneic lymphocytes.


Development of B lymphocytes. I. Cell populations and a critical event during ontogeny. The ontogeny of Ig(+), theta(+) and "null" (Ig-, theta-) lymphocytes in the mouse spleen was followed. An event was found at about 10 days after birth when the percentage of Ig(+) lymphocytes in the spleen begins to rise sharply, and the percentage of "null" lymphocytes falls. The Ig(+) lymphocytes before and after this event were then evaluated by four criteria pertaining to B cells. Three of these, the presence of C3 receptors, "capping" behavior, and the amount and/or disposition of surface Ig distinguished the Ig(+) lymphocytes before the event described from those present afterward. The fourth criterion, the presence of Fc receptors, did not distinguish between the early and mature forms of Ig(+) lymphocytes.


Rejection of mammary adenocarcinoma cell tumors in DBA/2 mice immunocompromised by thymectomy and treatment with antithymocyte serum. Syngeneic mammary adenocarcinoma cells were mixed with  killed Corynebacterium parvum organisms and were then transplanted subcutaneously into groups of normal and immunocompromised mice. The tumors formed at the site of injection exhibited normal growth for approximately 12 days followed by rapid and lasting rejection in both normal and immunocompromised animals. The control mice were protected against reinjections of 10-4 to 10-8 cells of the same line of tumor cells. The immunocompromised mice exhibited protection to reinjection of 10-4 cells of the tumor line initially employed. Microscopic examination of histologic sections of 10-day-old tumors in immunocompromised mice arising from administration of tumor cells and C.parvum mixtures revealed infiltration of only macrophages whereas those from normal animals exhibited both macrophages and lymphocytes. The data suggested that primary rejection of these tumors in immunocompromised mice may be the result of macrophage activity together with humoral immunity. These studies also revealed that 59 to 75% of immunocompromised mice exhibit protection against reinjection of the same line of tumor cells if twice-weekly treatment (i.p.) with killed C. parvum was included after inoculation of tumor cells mixed with C. parvum vaccine. These mice, unlike their saline-treated peers, exhibit delayed hypersensitivity reactions which are associated with proliferation of theta antigen-bearing lymphocytes. They also have intact tumor-directed humoral immune reactions.


Pathogenesis of rabies in immunodeficient mice. The HEP and ts2 strains of rabies virus inoculated intracerebrally into adult mice normally cause clinically inapparent infection. In the experiments described, this is converted into a lethal infection by immunosuppression with cyclophosphamide, which also prevented induction of immunity with vaccine. Lethal infection of HEP-inoculated mice was also observed in mice treated with antihymocytic serum, and in athymic (BALB/c-nu) nude mice.


Immunoglobulin biosynthesis in myeloma-associated and casein- and endotoxin-induced murine amyloidosis. Amyloidosis was induced in C57BL mice by daily injections of casein and in BALB/c mice by daily injections of endotoxin. There was no obvious disorder of immunoglobulin biosynthesis by spleen lymphocytes in these mice either before, during the development of, or in the amyloidotic stage. The pattern of immunoglobulin synthesis, assembly, and secretion was unaltered, the relative amount of heavy and light chains produced was normal, and there was an absence of immunoglobulin polypeptide chain fragments. Small amounts of amyloid were present in only 1 of 19 BALB/c and C3H mice (the IgG2a producing MOPC 173 tumor) bearing immunoglobulin-producing myeloma tumors and variants of these tumors. There was no relationship between excess light chain production by tumor plasma cells or spleen lymphocytes and the development of amyloidosis and there were no light chain fragments demonstrable. Antiserum prepared against casein-induced amyloid cross-reacted by immunofluorescence with the amyloid present in the MOPC 173 tumor-bearing mice, indicating the presence of common antigenic determinants in these two forms of amyloid. Attempts to study the biosynthesis of amyloid with incorporation of radioactively labeled amino acids were unsucessful.


Protective studies with a group A streptococcal M protein vaccine. II. Challange of volenteers after local immunization in the upper respiratory tract. Twenty-one adult volunteers were immunized at monthly intervals with three doses of purified type 1 M protein of group A Streptococcus. The soluble vaccine in buffer was administered by aerosol spray into the nares and oropharynx; 23 control subjects received a buffer placebo in the same manner. Antibody responses were observed in sera and nasal washings of some but not all vaccines. Approximately 30 days after  the last dose,  all subjects were challenged with homologus streptococci applied by swab to the phayngeal-tonsillar areas. In a double-blind system of evaluation, physical signs and symptoms were followed for assessment of infection. Illness was  defined on the basis of a positive throat culture, fever, a twofold increase in white blood cell count over baseline, exudative pharyngitis, and cervical adenopathy. By these criteria four vaccinees and 11 controls were obviously ill. One vaccinee and six controls were questionably ill, fulfilling some but not all of the criteria. sixteen vaccinees and six controls were not ill (P less than 0.001). Positive throat cultures were observed in five vaccines and 19 controls (P less than 0.001). Penicillin was administered five days after challenge. No poststreptoccal sequelae or other complication were observed. Thus local immunization with M protein apparently can prevent both colonization and clinical illness after challenge with homologous streptococci.


The diffusion and concentration of trimethoprim in human vaginal fluid. Colonization of the vaginal introitus with Enterobacteriaceae was cleared dramatically in women receiving trimethoprim-sulfamethoxazole for recurrent urinary tract infections. Vaginal fluid was collected from seven women; five of the seven had two vaginal fluid collections each while on full-dose therapy (total, 10 samples), and five had two or more collections each while on prophylactic, low-dose therapy (total, 14 samples). Bactericidal concentrations of trimethoprim were found in all samples of vaginal fluid, often exceeding the serum level by severalfold. Sulfamethoxazole was either undetectable or present only in fractional concentrations.


Disseminated candidiasis. i. an experimental model in the guinea pig. A model of experimental disseminated candidiasis was developed in the guinea pig; Candida albicans was injected intraperitoneally or intravenously. The kidney was the most severely affected organ, with maximal colony counts of 26,208 plus or minus 9,152 colony-forming units/g of tissue five days after a sublethal inocluation (one-sixth of the 50% lethal dose) of viable organisms. The heart was the next most severely affected organ, and other organs were affected little. Histologic studies confirmed the data from colony counts and showed a progression of inflammation similar to that seen in human disease. A leukocytosis predominantly of mononuclear cells, and to a lesser degree of granulocytes, occurred and was maximal four days after inoculation. This model will provide the framework for a series of future studies of the various factors important in natural host defense against infection with Candida, as well as the effects of various immunosuppressive agents, particularly corticosteroids, on the development and expression of immunity and protection against infection.


Isolation and characterization of Gal E mutant Ty 21a of Salmonella typhi: a candidate strain for a live, oral typhoid vaccine. A gal E mutant of Salmonella typhi was isolated; results obtained with Salmonella typhimurium and the mouse as a model for human typhoid fever indicated that this mutant has the potential for use as a live, oral typhoid vaccine. The mutant, Ty 21a, took up galactose from exogenous sources and accumulated sufficient quantities of galactose-1-phosphate and uridine diphosphate galactose to cause lysis of the cells, an event that resulted in the avirulence of the strain. Galactose was incorporated sufficiently into the cell wall of Ty 21a to allow the synthesis of smooth-type lipopolysaccharides, which are necessary for the proper immunogenicity. Cells of strain Ty 21a, when given intraperitoneally, protected mice against lethal challenge with strain Ty 2 of S. typhi.


Production and characterization of exotoxin(s) of Shigella dysenteriae type 1. A semicontinuous fermenter system was developed in which broth culture filtrates of Shigella dysenteriae type 1 yielded substantial amounts of exotoxin. Biologic activity of the exotoxin was characterized by means of three assays: the rabbit ileal loop for fluid evocation (enterotoxicity), mouse lethality after parenteral injection (neurotoxicity), and HeLa cell toxicity in vitro (cytotoxicity). Although the culture filtrate was highly active, disc electrophoresis revealed that the toxin is a minor component of the mixture of proteins in the crude preparation, and that the minor representation contrasts with the relative prominence of exotoxins in cultures of other bacteria, such as Vibrio cholerae, Corynebacterium diphtheriae, and Clostridium tetani. Filtrate toxin, when purified by isoelectric focusing in polyacrylamide gel, was found to contain two toxic moieties. One was resolvable as a single band with an isoelectric joint (pI) of 7.25, a molecular weight of 72,000, and all three types of biological activity. The second moiety was isoelectric at pH 6.00, contained two subcomponents, and further contrasted with the pI 7.25 toxin by being more cytotoxic while being devoid of enteroneurotoxic activity.


Involvement of the upper respiratory tract in orally induced salmonellosis in mice. The possible involvement of the tissues of the head and neck during the early pathogenesis of orally or intragastrically induced murine salmonellosis was examined. Viable counts of Salmonella in the local lymphoid tissues and draining nodes indicated that the ileal Peyer's patches were primarily involved in the development of a subsequent enteric fever, irrespective of the method of oral challenge. Contamination of drinking water with a high concentration of salmonellae produced infection of the ileal Peyers patches and the eventual development of systemic disease. The cervical lymph nodes were sporadically involved early in the infection, but this involvement soon became general, with large numbers of salmonellae in the cervical nodes by 36 hr. A lower concentration of salmonellae in drinking water led to primary involvement of the ileal Peyers patches; upper respiratory tract involvement was detected much later in the infection. Intragastrically invected mice exhibited the same low-level infection of the cervical lymph nodes as that in mice infected via drinking water. Thus systemic infection apparently results from infection of the ileal Peyers patches; a small percentage of the animals develop a concurrent upper respiratory tract infection that may be responsible for the development of carrier states after enteric infection.


Intracellular bactericidal activity of leukocytes in whole blood for the diagnosis of chronic granulomatous disease of childhood. A simplified method for the assay fo intracellular bactericidal activity of leukocytes in whole blood was developed. Providing that serum-resistant organisms are studied, the method can be used with gram-negative baccilli as well as with Staphylococcus aureus. The abnormalities characteristic of patients with chronic granulomatous disease of childhood and their mothers can be readily detected by this technique. By this means an abnormality in intracellular killing of Pseudomonas cepacia in a patient with chronic granulomatous disease and in his heterozygotic mother was found.


Transient bacteremia associated with percutaneous liver biopsy. Transient bacteremia associated with percutaneous liver biopsy was studied by pour-plate blood cultures, which were obtained immediately before and after the procedure and 5, 10, 15, and 30 min later in 89 patients. Part of the liver tissue was also cultured in all patients. Histological diagnoses included hepatitis, cirrhosis, cholangitis, fatty liver, granulomata, metastatic liver disease, lymphoma, and miscellaneous disorders. All blood cultures obtained before liver biopsy were sterile. Bacteremia was demonstrable in 12 patients (13.48%). In most of these patients, blood cultures were positive for as long as 15 min after liver biopsy; all cultures were negative at 30 min. Among the bacteria associated with 12 episodes of bacteremia were Escherichia coli, Klebsiella, Bacteroides, enterococci, diphtheroids, Staphylococcus aureus, alpha-hemolytic Streptococcus, and Streptococcus pneumoniae. The patients with positive liver biopsies had a higher incidence of bacteremia (83.3%) than did the patients whose liver biopsies were sterile (8.r%); this difference is stastically significant (P smaller than 0.01). Thus, liver biopsy can be associated with transient bactermia.


Nosocomial infection caused by gentamicin-resistant, streptomycin-sensitive Klebsiella. Resistance to gentamicin increased abruptly among nosocomial isolates of Klebsiella and Enterobacter at the New York Veterans Administration Hospital in 1973 and 1974. A prospective clinical survey revealed a greater incidence of true infection caused by Klebsiella than by Enterobacter. The initial site of implantation was usually the urinary tract. Multiple serotypes were involved, and one of these was found in rectal swabs of patients treated with antibiotics. Gentamicin resistance declined rapidly after the use of this antibiotic was restricted and increased several months after restrictions were removed. Gentamicin-resistant strains of Klebsiella were uniformly sensitive to amikacin; 75% were sensitive to polymyxin B and 70%  to streptomycin. Antibiotic sensitivity among gram-negative pathogens might be preserved by a program in which the use of highly effective agents is periodically rotated.


Antibody-coated bacteria in the urine of obstetrical patients with acute pyelonephritis. The direct immunofluorescence method for the detection of antibody-coated bacteria in urine sediments was used to test urine samples from obstetrical patients with the clinical diagnosis of acute pyelonephritis or cystitis. Antibody-coated bacteria were present in the urine from 12 of 15 patients with acute pyelonephritis, but they were not observed in the urine from 13 patients with cystitis. The clases of antibody coating the bacteria were IgG, IgA, and, in some cases, IgM. A correlation between a high titer of antibody in serum and the presence of antibody-coated bacteria in the urine was noted. These results confirm that the immunofluorescence test can be useful, as previously reported, in distinguishing infection of the kidney from infection of the bladder.


Meningitis caused by Streptococcus in adults. Meningitis caused by Streptococcus is uncommon in adults. Otitis media, mastoiditis, sinusitis, or trauma has antedated most cases reported in the past. Data on 10 recent cases suggest changes in the current pathogenesis of streptococcal meningitis in adults and emphasize the importance of accurate streptococcal speciation. Endocarditis was present in five cases; trauma played a role in two others. Brain abscess, corticosteroids, alcoholic cirrhosis, and peritonitis secondary to chronic peritoneal dialysis were etiologic factors in five patients. Otitis media, mastoiditis, and sinusitis were conspicuously absent. Streptococcus agalctiae accounted for purulent meningitis in two postmenopausal women. Both strains of group B Streptococcus were bacitracin-sensitive and were thus mislabeled group A. Recognition of Streptococcus bovis spared two patients unnecessary aminoglycoside adminstration. Nine of the 10 patients survived, including a women with prosthetic mitral valve endocarditis and a man with a ruptured brain abscess. Differentiation of these streptococci from Streptococcus pneumoniae was seldon possible on the basis of the spinal fluid gram stain alone.


Otology in the Bible and Talmud.

Otology in the Bible and Talmud. The present paper is an English translation of the chapter of Otology in Preuss's classic book Biblisch-Talmudische Medizin. Described in the Talmud are a variety of anatomical ear abnormalities such as double ear, pierced ears, small ears, cut off ears, and pendulous ears. A slave whose ear was wounded had to be freed. Compensation for injury to the ear and to hearing is clearly noted. Remedies for earache are mentioned. The legal status of a deaf-mute, and the relationship of deafness to muteness are also discussed in the Talmud.


Determination of bile acid pool size in man: a simplified method with advantages of increases precision, shortened analysis time, and decreased isotope exposure. A simplified isotope dilution method for measurement of the bile acid pool size in normal subjects is described and compared with the traditional method of Lindstedt (Acta Physiol. Scand. 40: 1-9, 1957). Advantages of this simplified method include a four- to eightfold reduction of isotope dose, facilitation of analytical procedures, and a reduction in the required number of duodenal intubations. In 15 human subjects who had two separate estimates of pool size by this method, precision averaged 2.6 percent. In 16 comparisons, pool size measured by this method averaged 13.7 percent higher than simultaneous estimates by the Lindstedt method. Factors affecting accuracy (as opposed to precision) in both methods are discussed.


Lyophilized 7 alpha-hydroxysteroid dehydrogenase: a stable enzyme preparation for routine bile acid analysis. Preparations of 7 alpha-hydroxysteroid dehydrogenase (7 alpha-HSDH) from Escherichia coli strain 23 can be frozen and thawed without significant loss of activity. 7 alpha-HSDH may then be lyophilized into powder form, which is stable for more than 6 months (3% loss of activity). The lyophilized 7 alpha-HSDH preparation has the additional advantage over previously described preparations of a low and stable fluorescence background when applied to the fluorometric estimation of bile acids, especially in combination with thin-layer chromatography. Analysis of duodenal aspirates from 18 normal subjects gave bile acid ratios identical with those reported earlier and obtained by using gas-liquid chromatography. A significant difference in the glycine: taurine ratio between males and females was observed.


Genetical control of B-cell responses. III. Requirement for functional mitogenicity of the antigen in thymus-independent specific responses. Spleen cells from C3H/HeJ mice fail to respond with polyclonal antibody synthesis to mitogenic concentrations of lipopolysaccharide (LPS) which are optimal for activating spleen cells from a high-responder strain (B10.5M). This unresponsiveness is selective for LPS, since C3H/HeJ cells respond as normals to another B-cell mitogen, purified protein derivative of tuberculin. Spleen cells from low-responder mice also fail to mount a specific anti-NNP plaque-forming cell (PFC) response, when challenged in vitro by NNP-LPS. However, C3H/HeJ cells develop normal responses to another thymus-independent hapten conjugate, DNP-AECM-Ficoll. C3H/HeJ mice fail to mount a specific anti-LPS antibody response, when challenged in vivo with doses of soluble LPS which are fully immunogenic for the high-responder strain. However, C3H/HeJ mice develop normal direct and indirect PFC responses to LPS, when challenged with a thymus-dependent form of the immunogen. These results are interpreted as indicating as absolute requirement for functional mitogenicity of the antigen, in the induction of specific thymus-independent antibody responses.


Immunologic injury of cultured cells infected with measles virus. I. role of IfG antibody and the alternative complement pathway. In these studies, a number of human cell lines including epithelial, neural, glial, and lymphoid cells infected with several strains of measles virus were found to be lysed upon incubation with fresh sera from humans containing antibody measles virus. In all instances, the cytolytic event was mediated by alternative complement (C) pathway without a significant contribution from classical pathway. In contrast, isolated measles virus in conjunction with antibody was found to selectively activate the classical C pathway. Measles antibodies of the IgG class, but not the IgA class, possessed cytolytic potential against cells infected with measles virus. Human IgG antibodies could directly activate the alternative C pathway. No defect was found in cytolytic measles antibody in sera or cerebrospinal fluid from patients with subacute sclerosing panencephalitis, nor was the alternative C pathway impaired in sera from these patients. Sera from newborn humans exhibited a functional alternative C pathway.


Study of the cells proliferating in parent versus F hybrid mixed lymphocyte culture. Caryotypic analysis of the cells dividing in mouse parent-hybrid MLC showed an F1 hybrid cell proliferation, which varied depending upon the source of lymphoid cells used: strong in spleen MLC (sometimes equal to that of the parental cells), less marked in lymph node cell MLC, and most often absent in MLC between cortisone-resistant (CR) thymocytes. MLC between parental spleen cells and F1 CR thymocytes showed, however, that in certain conditions of culture F thymocytes can also proliferate. Using parental or F1 spleen cells lacking T lymphocytes, it was found that F1 cell proliferation is entirely dependent upon the presence of parental T cells, but does not require the presence of T lymphocytes among the F1 cells. Immunofluorescence analysis of the blasts observed in one-way MLC showed that about 70% of the parental blasts were T blasts, and 25%B blasts (containing a high proportion of plasmablasts); among the F1 blasts, there was also the same percentage of B blasts and plasmablasts, but many of the T blasts bore only small amounts of T-cell antigen (MTLA), and there was also about 20%of unstained blasts, possibly T blasts bearing MTLA in amounts undetectable by immunofluorescence. The possibility is discussed that the F1 responding T cells belong to a subpopulation performing a suppressive function; MLC lacking F1 T cells showed increased [3H] thymidine incorporation. The proliferation and differentiation of parental and F1 B cells may result mainly from an unspecific, "polyclonal" triggering.


Effects on C-reactive protein on the lymphoid system. I. Binding to thymus-dependent lymphocytes and alteration of their functions. C-reactive protein (CRP) is an acute phase protein which shares with the immunoglobulins the ability to induce precipitation and agglutination reactions and activate the complement system. We report here that purified human CRP binds selectively to human T lymphocytes, inhibits their ability to form spontaneous rosettes with sheep erythrocytes and inhibits their response to allogeneic cells in mixed lymphocyte culture reactions; it fails to inhibit phytohemagglutinin- or concanavalin-A-induced mitogenesis. CRP does not bind to human B lymphocytes, nor does it alter the following B-cell functions: binding to activated complement components or the Fc portion of immunoglobulins, mediation of antibody-dependent cytotoxicity reactions or the ability of allogeneic cells to stimulate a mixed lymphocyte culture reaction. Human CRP shows similar selective binding with murine T lymphocytes. It therefore seems that binding of CRP is a property of T lymphocytes or a subpopulation thereof, and can result in modulation of certain of the T-cell functional characteristics in vitro. We suggest that CRP may play a role in modulating T-cell functions during the inflammatory state.


Elevated ribonuclease activity in the thymus and white blood cells of genetically cancer prone mice. Ribonuclease activity in cell-free thymus homogenates was elevated for five strains of mice genetically predisposed toward leukemia or reticulum cell neoplasms (AKR, C58, PL, RF, and SJL). Such increased activity was directed against polyuridylic acid and was observed in 8-wk old mice, well before the onset of neoplastic transformation. Similarly, white blood cell ribonuclease activity was elevated in mice of the strains AKR, C2H/He, PL and RF. Statistical analysis indicated that such elevated activity in these strains related to their high incidence of spontaneous neoplastic disease. Elevated ribonuclease activity thus represents a new biochemical marker relating to the genetic propensity of some strains of mice to die prematurely of spontaneous neoplasia.


Proliferation and colony-forming ability of peritoneal exudate cells in liquid culture. Peritoneal exudate cells, obtained from mice injected with thioglycollate medium and cultured in medium containing L-cell-conditioned medium, will proliferate in an exponential fashion for 18 days with a doubling time of 68 h. After a 2 h pulse of tritiated thymidine, labeled adherent cells increased to a maximum of 22-34% during the 1st and 2nd wk of culture. Increasing the cell concentration from 2 times 10-3 to 2 times 10-5 cells/culture reduced exponential growth to 10 days and the doubling time was increased to 81.6 h. Under these culture conditions, peritoneal exudate cells were shown to form colonies on the surface of culture dishes when plated at low density. The cells within the colony were shown to be macrophages using yeast and antibody-coated sheep erythrocytes as a test for phagocytic function. The plating efficiolonies arose from a single precursor cell. The adherent cell population contains the colony-forming precursors. These precursors can be stimulated to form colonies for at least 2 wk by the addition of conditioned medium to cultures at various times after plating. While very few colony-forming cells could be demonstrated in the unstimulated peritoneal lavage, their numbers begin to increase in the exudate 4 h after injection of thioglycollate medium and reach a maximum by day 3 and then decrease. Isolated colonies may be useful in studying the function of macrophages.


T-cell migration into allografts.

T-cell migration into allografts. The ability of T and B lymphocytes to migrate into skin allografts undergoing rejection was studied in mice. Spleen cells from CBA/J mice sensitized to transplantation antigens of A/J or C57BL/6 mice were separated on immunabsorbent columns into purified populations of T and B cells, labeled in vitro with 3H-uridine and injected intravenously into CBA/J mice with 7-day old skin iso and allografts (A/J or C57BL/6). The mice were sacrificed 24 h later and studied by autoradiography. After transfer of either unfractionated spleen cells or T cells, large numbers of labeled cells were found in the cellular infiltrate of allografts, whereas extremely few were seen in isografts. In contrast, after transfer of B cells, almost no labeled cells were detected either in the allografts or the isografts, although they, like T cells, homed normally to lymphoid tissue.


Rubella cataract in vitro: Sensitive period of the developing human lens. The clinically known sensitive period of rubella cataract was studied in vitro by infecting 79 human eye rudiments from embryos aged 4-10 wk with rubella virus. The course of the infection was followed by histological and indirect immunofluorescence methods. Of the rudiments, 12 pairs were in the lens placode or open-lens-vesicle stage, 40 already had closed lens vesicles and in another 27 closed-stage pairs an incision was made in the lens capsule before infection to allow the virus to enter the lens. Uninfected controls differentiated well in vitro for 4-6 wk. The eye rudiments infected in the open-lens-vesicle stage showed lens fiber destruction and viral antigens within the lens. No damage or viral antigens were detected in rudiments infected in the closed stage unless the lens capsule was incisedmwhen this was done, however, fiber damage ensued and viral antigens appeared. The lens capsule was concluded to form a protective barrier around the sensirive fibers at the time of closure of the lens vesicle, confirming the earlier hypothesis and clinical findings.


Characterization of the macrophage receptro for complement and demonstration of its functional independence from the receptor for the Fc portion of immunoglobulin G. The complement receptor of the macrophage membrane recognizes particle-bound C3b but does not recognize particle-bound C3d. C3-b-coated sheep erythrocytes were bound to macrophages via their C3b receptors, and the preparations were then incubated with either latex particles or opsonized pneumococci (test particles). Macrophages ingested the test particles, but erythrocytes were not ingested; they remained bound to C3b receptors of the macrophage plasma membrane. Thus, a signal initiating ingestion via one type of receptor is not transmitted to all receptors which have the potential to mediate phagocytosis.


Clonal dominance. I. Restricted nature of the IgM antibody response to group A streptococcal carbohydrate in mice. The IgM antibody response of mice to the streptococcal group A carbohydrate (GAC) was measured. With most strains tested, large amounts of IgM antibody were produced; in AKR mice, over 1% of the total nucleated spleen cells secreted IgM anti-GAC antibody after hyperimmunization. The relative avidity of the antibody was extimated by a modification of the Jerne plaque assay where spleen cells from individual mice were tested against erythrocytes with varying GAC epitope densitymthese studies showed that the earliest, as well as latest, IgM antibodies produced were highly restricted in avidity heterogeneity. No evidence of affinity maturation was seen upon hyperimmunization. These data favor the conclusion that the restricted IgG response seen in mice hyperimmunized to GAC is not the result of affinity driven competition for antigen among precursor cells.


Functional subclasses of T-lymphocytes bearing different Ly antigens. I. The generation of functionally distinct T-cell subclasses is a differentiative process independent of antigen. Ly alloantigens coded by two unlinked genetic loci (Ly-1 and Ly-2/Ly-3) are expressed on lymphoid cells undergoing thymus-dependent differentiation. Peripheral Thy-1+ cells from C57BL/6 mice can be divided into three subclasses on the basis of differential expression of Ly-1, Ly-2, and Ly-3; about 50% express all three Ly antigens (Ly -123+), about 33% only Ly-1 (Ly-1+), and about 6-8% Ly-2 and Ly-3 (Ly-23+). Cells of the Ly-123+ subclasses are the first peripheral Thy-1+ cells to appear in ontogeny, and are reduced in the periphery shortly after adult thymectomy. In contrast, Ly-1+ and Ly-23+ subclasses appear later in the peripheral tissues than do Ly-123+ cells, and are resistant to the early effects of adult thymectomymperiheral lymphoid populations depleted of Ly-1+ cells and Ly-123+ cells (and thereby enriched for Ly-23+ cells) were incapable of developing significant helper activity to SRBC but generated substantial levels of cytotoxic activity to allogeneic target cells. The same lymphoid populations, depleted of Ly-23+ cells and Ly-123+ cells (and thereby enriched for Ly-1+ cells), produced substantial helper responses but were unable to generate appreciable levels of killer activity. These experiments imply that commitment of T cells to participate exclusively in either helper or cytotoxic function is a differentiative process that takes place before they encounter antigen, and is accompanied by exclusion of different Ly groups, Lu-23 or Ly-1 respectively, from TL+Ly-123+ T-cell precursors. It is yet to be decided whether the TL-phase by Ly-123+ subclass is a transitional form or a separately differentiated subclass with a discrete immunologic function.


Functional subclasses of T lymphocytes bearing different Ly antigens. II. Cooperation between subclasses of Ly+ cells in the generation of killer activity. Lymphocytes from BALB/c or C57B6/6 mice that develop killer activity to alloantigens belong to the numerically small Ly-23 subclass of peripheral T cells, distinguished by selective expression of Ly-23 determinants on their surfaces. The maturation of these cells to killer cells can be amplified by Ly-1+ cells, which do not themselves contribute to the killer cell pool. This amplification was abolished by escluding Ia("Beta")+ cells from the stimulator population during mixed lymphocyte culture (MCL), suggesting that amplification is due to selective recognition of I region antigens by L-1+ cells, a conclusion already drawn from our previous evidence that Ia differences activate Ly-1+ cells but not Ly-23+ cells. These and other experiments indicate that amplification of killer cell production in vitro by Ly-1+ cells is not due to their conversion to Ly-23+ cells during MLC, but to their ability to recognize major histocompatibility complex determinants not recognized by Ly-23+ cells.


Office bacteriology in children.

Office bacteriology in children. Office bacteriological procedures can be a valuable part of a physician's diagnostic armamentarium when utilized within the framework of sound clinical judgment. This is particularily true in children, where infectious diseases represent the major portion of illness. The techniques are simple and inexpensive. The expertise necessary is rapidly and easily acquired. The results can be gratifying and serve as a stimulant for greater involvement in this technique.


Recent advances in diagnosis and management of croup. Croup is a syndrome of inspiratory stridor, cough, and hoarseness, due to varying degrees of laryngeal obstruction. It is a viral disease and must be differentiated from epiglottitis. In addition to a careful clinical assessment, neck films are the most valuable diagnostic tool in differentiating these two. The principle modes of therapy for croup include provision of adequate hydration, ensuring maximum available humidification, sedation, and administration of intermittent positive pressure breathing (IPPB) with nebulized racemic epinephrine. The latter mode of therapy will provide symptomatic relief and may eliminate the need for hospitalization and tracheostomy. Steroids offer little benefit in treating this disease, and antibiotics offer none.


The effects of castration and androgen replacement on song, courtship, and aggression in zebra finches (Poephila guttata). Castration of adult male zebra finches (Poephila guttata, Estrildidae) reduces their singing rate and the tempo of song, but castrates continue to sing song identical in form to preoperative song. Injection or implantation of testosterone propionate (TP) but not of vehicle alone reverses the changes produced by castration. Castration or partial castration also reduces the frequency of courtship, copulation, and aggression. Androgen (TP) replacement reverses these changes, but control injections do not. The persistence of song after castration contrasts with the abolition of song by castration in other birds, and this may be related to the natural history of zebra finches.


A method for the chemical isolation of individual muscle fibers and its application to a study of the effect of denervation on the number of nuclei per muscle fiber. A method is described for isolating intact, individual skeletal muscle fibers from glutaraldehyde fixed muscle. This method was conceived to climinate the many limitations of determining muscle nuclear numbers in histological cross section. Glutaraldehyde fixed fibers are isolated by dissection while in a solution of low concentration guanidine in a borate buffer at high pH. Electron miscroscopy demonstrates that single fibers, isolated in this manner, are free of their microvasculature and connective tissue. Their basal laminas and the structures within them, including their satellite cells, are preserved. This method is employed to determine whether changes in nuclear numbers occur within the basal lamina of individual muscle cells from 1 to 28 days following denervation of mouse gastrocnemius muscle. The total number of nuclei located within the basal lamina of individual muscle fibers (i.e. muscle and satellite cell nuclei) does not change after denervation. This rules out the possibility that additional nuclei are arising from an influx of cells outside the basal lamina or by mitotic division of nuclei within the basal lamina. However, the possibility of a change in the ratio of satellite cell nuclei, to muscle cell nuclei, is not eliminated. Other possible applications of this isolation method are discussed.


The immune response of the teleost, Tilapia mossambica to soluble and cellular antigens. The immune response of Tilapia mossambica to bovine serum albumin (BSA) and sheep red blood cells (SRBC) was characterized in detail in terms of the appearance of hemolysin plaque-forming cells and circulating antibodies at 30 degrees C. Plaque-forming cells (PFC) were detected in the spleen, head-kidney and thymus of immunized fish and the maximum number was observed in these organs on the fifth day after immunization with SRBC. Peak circulating antibody response occurred on day 8 for SRBC and on day 11 for BSA. Following the second injection of the same antigen, a specific anamnestic response was observed with increased production of PFC and serum antibody. No cross reactivity was found when anti-SRBC antibody was tested with rat erythrocytes. Tests with 2-mercaptoethanol showed that all of the agglutinating antibody produced after both the first and second injection was mercaptoethanol sensitive. Analysis of histological and smear preparations revealed that there were consistent cellular changes occurring in the spleen as well as the head-kidney due to immunization.


Induction of nuclear breakdown and meiosis in Spisula solidissima oocytes by calcium ionophore. Induction of germinal vesicle breakdown and reinitiation of the meiotic process resulted following exposure of immature Spisula solidissima oocytes to divalent ionophore A23187. Meiosis continued to completion in the presence of the ionophore; however, parthenogenetic development of the gametes did not occur. Incubation of oocytes in calcium deficient media (Ca++ free or Ca++Mg++ free seawater) but not Mg++ free seawater inhibited ionophore induced maturation. Effects of calcium deficiency were reversible. The results suggest that nuclear disintegration and the meiotic process induced by sperm are mediated by ionic stimuli and that the establishment of meiotic arrest is linked to the availability of calcium within the oocyte.