Immune studies in oral contraceptive users. Some of the parameters of immune status-percentage of B and T lymphocyte subpopulation, PHA-induced lymphocyte transformation measured by 3H-thymidine incorporation (PILT) and levels of total haemolytic complement (CH5O) were studied in Indian women from low income group who were using estrogen-progestogen combination pills. There were no differences in percentage of B and T lymphocyte subpopulation or PILT between OC users and the control group. However, CH5O levels were significantly lower in OC users. The depression in circulating complement levels in OC users is intriguing and suggests the possibility that complement system and circulating immune complexes may be altered in OC users.


Drugs on a medical campus I. Drug use among medical undergraduates. All 750 undergraduate students of the Medical College, Patiala, during the academic year 1976--1977 were covered in this survey. Seven out of ten students were found to have used some drug in the past. The lifetime prevalence being 82.4% among boys and 29.6% among girls. A majority of the students (78.6%) were poly-drug users, the commonest drugs used being alcohol (58%), tranquillizers (47%), and tobacco (36%). However, the prevalence of daily use was low, it being highest for cigarette smoking, by 9.4% of students, while less than 2% reported daily use of alcohol. The only other drugs used on a daily basis were stimulants (0.9%), tranquillizers (0.6%), and sedatives (0.3%). Of the opium users 81.8%, and over 60% of alcohol and tobacco users, had started taking these drugs during high school or the pre-medical year. On the other hand a majority of the students using tranquillizers (75%) and stimulants (66%) began using these drugs after entering the Medical College. The percentage of drug users increases by approximately 10% during each year of their medical studies.


[Acute bacterial endocarditis caused by streptococci of Lancefield group C (author's transl)]. An Angell-Shiley heterograft valve was implanted in a 32-year-old woman with severe aortic regurgitation and stenosis. Post-operatively acute bacterial endocarditis occurred, due to group C streptococci. Because of severe acute aortic insufficiency with partial dehiscence of the heterograft valve and increasing left heart failure re-operation was necessary. After successful replacement of the valve an aortocoronary bypass was connected to the left circumflex artery because of displacement of the left coronary ostium. In addition, an aortoplasty was performed for spontaneous aortic rupture. Because of severe left heart failure with myocardial infarction in the course of the operation circulatory support with a paracorporeal artificial heart was necessary for 60 hours postoperatively. Despite transitory improvement the patient died from septic shock 30 days after the re-operation.


Effect of dopamine-beta-hydroxylase inhibitor (disulfiram) on the response of adenohypophysis, serum ceruloplasmin and hypothalamic ascorbic acid to estradiol treatment. The administration of tetraethylthiuramdisulfide (disulfiram, Antabus--inhibitor of dopamine-beta-hydroxylase) resulted in an inhibition of the response of anterior pituitary to estradiol (i. e. increase of weight and of binding capacity of anterior pituitary proteins to thyroxine). At the same time there was no inhibition of an increase of ceruloplasmin (polyphenoloxidase) level after estradiol, while a decrease of ascorbic acid in hypothalamus was fully prevented. It was concluded that disulfiram may inhibit dopamine-beta-hydroxylase activity and thus presumably protect the hypothalamic dopamine which further inhibits the response of anterior pituitary to estradiol either directly or through a dopamine dependent growth inhibiting factor.


Natriuretic and kaliuretic effect of melanocyte-stimulating hormones in hamsters. The effect of synthetic melanocyte-stimulating hormones (MSH) on urinary excretion of sodiu, potassium and water was studied in hamsters. Synthetic alpha-MSH and synthetic human beta-MSH showed a marked natriuretic and diuretic effect depending on the dosage of hormone. The natriuretic effect of both hormones was approximately equal and did not parallel the magnitude of the melanocyte-stimulating activity of the investigated peptides. Both peptides showed also a milder kaliuretic effect independent on their dose. The rise of the sodium/potassium index in urine was significant after both peptides. Bilateral adrenalectomy increased further the natriuresis induced by alpha-MSH, while it did not affect the effect o MSH on diuresis and kaliuresis. Changes in urinary excretion of sodium and potassium are considered to be consequence of direct renal action of MSH.


Preferential uptake of estradiol in the pituitary and hypothalamus of mature male rats in the presence of testosterone. Simultaneous intravenous injection of 3H-estradiol and 14C-testosterone into adult male rats resulted in significant concentration of both labels in the blood and the diencephalon 2 h after the injection. In the pituitary--hypothalamic area only estradiol derived radioactivity could be detected, while testosterone derived 14C label was not found. The results are interpreted to indicate that in males estradiol, not testosterone, may provide the homeostatic balance between the gonads and the hypothalamo-pituitary axis.


Diminution of prostate by ergoline derivative VUFB--6638 (Dironyl) in rats. Experiments in male rats showed that the administration of prolactin secretion inhibitor-N-(D-6-methyl-8-isoergolin-I-yl)-N', N'-diethylurea -- led to a decrease of prostate weight, while a simultaneous administration of prolactin prevented this effect. The histological examination showed that the quantity of secretions in the prostate glands are decreased.


Effect of deamino-dicarba-oxytocin and oxytocin on myoelectrical and mechanical activity of uterus, stomach and small intestine in dog. In conscious dog deamino-dicarba-oxytocin strongly stimulated a spike activity in the uterus which was more prolonged than that after oxytocin. Both oxytocin and deamino-dicarba-oxytocin inhibited the tone and peristaltic contractions of stomach and small intestines in starving dogs, eliminated the spike discharges, reduced the basic electrical rhythm frequency of the stomach and accelerated the propagation velocity of the slow waves in the small intestine. Only oxytocin, but not its deamino-analogue, reduced the pulse frequency and inhibited the spontaneous contractions of muscle strips isolated from the guinea pig stomach and portal vein.


alpha 1-Antitrypsin deficiency and susceptibility to lung disease. This subject concerns the complex interrelationship of a genetically determined protein deficiency, enzymes which are inhibited by that protein, environmental challenges such as cigarette smoke and industrial pollutants, and the occurrence of obstructive lung disease (Fig. 1). Unequivocal establishment of an etiological role for AAT deficiency, especially of intermediate degree, has proven to be difficult. Confounding variables such as enzyme concentration in PMN and PAMs, duration of exposure to potential environmental hazards, differences in laboratory methods utilized in measuring AAT and in studying pulmonary function all require investigation. The definitive study, incorporating all of these and other factors, has yet to be conducted. No single, clear-cut conclusion can be drawn from analysis of present studies. In those circumstances in which heterozygotes appear to be predisposed to COPD, phenotypic screening of the population at potential risk, such as industrial workers may be appropriate. Conversely, in conditions in which no association is demonstrated, such screening would not be justified. Perhaps, the best one can do is to suggest a "Scotch verdict"; that is, the issue of causation is not proven.


Mitral valve replacement in infective endocarditis as prophylaxis against embolism. Identification of patients at risk by 2-dimensional echocardiography. A 36-yr-old woman and a 46-yr-old man had infective endocarditis of the mitral valve. Examination by 2-dimensional dynamic echocardiography demonstrated large mobile vegetations in both patients, and the display pointed to an ominous risk of embolism. Heart surgery with valve replacement was performed, the main indication being prophylaxis against embolism; the operative findings seemed to justify the assumption of imminent risk of embolism. It is suggested that the display of dynamic morphology of valvular vegetations by 2-dimensional echocardiography can be useful in identifying a subset of patients at high risk of embolism during infective endocarditis. However, more experience is needed before definite conclusions can be drawn regarding the role of early operation as prophylaxis against impending embolism.


Cytotoxic T cell responses to haptenated cells. I. Primary, secondary and long-term cultures. Cytotoxic T lymphocyte (CTL) responses were obtained in vitro to cells coupled with several different haptens. The degree of lysis of target cells was dependent on the amount of hapten coupled to stimulator and target cells. Spleen cells from normal mice responded to high-hapten density cells but not to low-hapten density cells. However, spleen cells from immunized CBA mice could be stimulated in vitro by low-hapten density cells to generate effector cells able to lyse low-hapten density cells. In vitro primed responder cells could be restimulated in the presence of the original hapten-coupled stimulator cells or in the presence of supernatant from concanavalin A-stimulated mouse or rat spleen cells. Large number of hapten-specific and H-2-restricted CTL could be generated by repeated exposure to fresh supernatant.


Studies on the generation and expression of H-2-controlled T helper function in chimeric mice: evidence for two levels of H-2 resitriction. F1 leads to parental, semisyngeneic/semiallogeneic and fully allogeneic bone marrow radiation chimeras were used as a source of helper T cells in the in vitro anamnestic response to dinitrophenylated keyhole limpet hemocyanin. In F1 leads to parental and in semisyngeneic/semiallogeneic chimeras, a small population of helper T cells restricted to the H-2 haplotype present in the donor only was shown to co-exist with T cells restricted to the shared haplotype. Only the population restricted to the donor H-2 could be demonstrated in allochimeras, presumably due to a lack of antigen-presenting cells of host origin during priming. Under assay conditions where, in addition to T cell/macrophage interactions, a direct T-B cell contact was necessary ("linked" cooperation), the H-2 restrictions observed were absolute. When direct T-B cell contact was made unnecessary by a special experimental protocol ("unlinked" cooperation), H-2 restriction between T and B cells was overcome. Thus, the cellular interactions during the anamnestic immune response to T-dependent antigens seem to be H-2-restricted at two levels: T cell/macrophage and T-B cell interactions.


Hairy cell leukemia: effect of 'hairy' cells on normal granulopoiesis in vitro. The effect of 'hairy cells' (HC) on proliferation of normal marrow granulocytic progenitor cells (CFU-C) was studied in vitro, using cells obtained from the blood, spleen, and bone marrow from eight patients with hairy cell leukemia (HCL). Hairy cells of hairy-cell-conditioned media (HCCM) did not stimulate proliferation of normal CFU-C. Furthermore, HC and HCCM did not inhibit the growth of CFU-C induced by normal colony stimulating factor (CSF) or by feeder layers of normal blood leukocytes. These results confirm previous reports showing the inability of HC to produce CSF and suggest that inhibition of CFU-C proliferation, as measured by this technique, is not an important mechanism in the neutropenia observed in HCL.


Stimulation of thymocytes in vitro by serum of 2-acetylaminofluorene (AAF)-treated rats. The serum of rats obtained at various time intervals after administration of AAF was tested for its action on the 3H-thymidine incorporation into the DNA of thymocytes in vitro. In comparison with untreated or solvent-treated controls, the sera of animals in the early phase after the AAF-treatment stimulate the incorporation of the nucleotide into the thymocytes of rats, mice and hamsters. This reaction shows an unequivocal dose-response relationship. The serum activity is dialysable and relatively thermostable. Spleen lymphocytes and peritoneal cells of rats are insensitve to the AAF-serum. A possible relationship between growth-stimulating serum activities in liver regeneration and in early carcinogen action is discussed.


Some ultrastructural observations on the denervated skeletal muscle of frog. The frog skeletal muscle resists atrophy for a long time after denervation. If, however, the entry of the nerve is prevented for more than 3.5 months, small muscle portions, sometimes containing nucleus, are sequestered from the parent muscle fibre. The basal lamina does not dissociate from these detached muscle portions.


Suppression of intrinsic B-cell function in Dengue-infected mice. Mice infected with Dengue virus show a depressed immune response to lipopolysaccharide (LPS), a helper T-cell-independent antigen, when LPS was administered on day 0, 6 and 12 post infection. Mice injected with inactivated virus failed to show immunosuppression.


Kallikrein inhibitors.

Kallikrein inhibitors. So far the Cl inactivator, alpha 2-macroglobulin, antithrombin III (in the presence of heparin), and alpha 1-antitrypsin have been identified as inhibitors of plasma kallikrein; alpha 1-antitrypsin reacts slowly also with tissue kallikreins. Of the various naturally occurring kallikrein inhibitors the basic trypsin-kallikrein inhibitor of bovine organs, aprotinin (the active substance of Trasylol), has attained by far the most interest. This inhibitor, which is produced by mast cells, has unusual properties due to its compact tertiary structure. Additional topics of aprotinin and structurally related inhibitors discussed are the mechanism of enzyme-inhibitor complex formation, the production of chemical mutants of aprotinin, the structural basis of kallikrein inhibition, and selected aspects regarding aprotinin medication.


[Effect of peptide hormones of the gastrointestinal tract on leukocytes]. A single i. m. administration of pentagastrin or secretin changed the amount of lymphoid and myeloid cells in rats and mice. A total decrease of leucocytes is due to decrease of amount of lymphocytes, eosinophiles, and monocytes. Simultaneously the amount of segment-nuclear and relating to stab neutrophiles increased. After pentagastrin administration the amount of rosette-forming T-lymphocytes decreases. The data obtained suggest the effect of gastrin and secretin on the blood-forming and blood-destroying systems of the organism.


Mesenteric varices: a source of mesosystemic shunts and gastrointestinal hemorrhage. The presence of mesenteric varices was demonstrated angiographically in 7 patients with portal hypertension. In 4 of these cases the mesenteric varices were the source of lower gastrointestinal bleeding which was successfully controlled by intra-arterial infusion of vasopressin. The radiological diagnosis and management of mesenteric varices is discussed and the pertinent literature is briefly reviewed.


[Intensive care medicine in severe eclampsia (author's transl)]. The intricacies of the pathophysiology of eclampsia are still unknown. The major symptoms of our 37 year old para 3 are convulsions, hypertension, complete anuria and gastro-intestinal haemmorhage as a result of disseminated ;ntra-vascular coagulation (D.I.C.). The interdisciplinary therapeutic measures are discussed, in the course which special attention is given to the favourable influence of dopamine on renal failure and the complicating gastro intestinal haemmorhage.


Adhesion of blood platelets to human arterial subendothelium: role of factor VIII-Von Willebrand factor. The adhesion of human platelets to the human arterial subendothelium was investigated using everted postmortem renal arteries after the first bifurcation and reconstituted blood containing 51Cr-labelled, aspirin-treated platelets in a perfusion chamber according to Baumgartner. The accumulation of 51Cr on the arterial segment was a reliable reflection of the number of platelets that adhered. The dependence of platelet adhesion on platelet number, perfusion time, haematocrit and perfusion rate were found to be similar to what was observed with rabbit aorta previously, notwithstanding the essentially different nature of the human subendothelium, with its layers of smooth muscle cells above the internal elastic lamina. The system was used to study the role of factor VIII-von Willebrand factor (VIII-VWF). VIII-VWF was found to be the only plasma factor enhancing adhesion of platelets fo the vessel wall. No suggestion of an inhibitory substance in plasma was found. Double perfusion experiments indicated that VIII-VWF binding to subendothelium supported subsequent platelet adhesion. A good correlation was found between the amount of VIII-VWF bound and platelet adhesion. Comparison of binding of albumin, fibrinogen, gamma-globulin and VIII-VWF showed that VIII-VWF bound less. This suggests that VIII-VWF does penetrate less easily in the subendothelium, which is in agreement with immunofluorescence studies of tissues. Studies with washed platelets not treated with aspirin showed parallelism between accumulation of VIII-VWF and platelet deposition in thrombi. These data may be explained by assuming a non-easily exchangeable pool of VIII-VWF on the platelet surface and increased binding of VIII-VWF to platelets in thrombi.


PiT: a new allele in the alpha 1-antitrypsin system. In a genetic investigation of the population in Hessen, Germany, we found a family with a new, rare allele in the Pi system (alpha 1-antitrypsin). According to electrophoretic analysis and isoelectric focusing patterns, it is designated PiT. A pedigree study suggests autosomal codominant inheritance. The serum concentration of six heterozygous carriers of this allele (phenotype M1T or M2T) revealed normal alpha 1-antitrypsin levels.


Human pancreatic amylase polymorphism: formal genetics and population genetics. The genetically determined polymorphism of human pancreatic amylase (E.C. 3.2.1.1), AMY2, is demonstrated in serum specimens by agarose gel electrophoresis. We investigated 325 mother-child pairs and 2594 unrelated individuals from southwestern Germany. This study confirms the formal hypothesis of two common alleles AMY1/2, AMY2/2 and possibly two rare alleles AMY3/2, AMY4/2 at an autosomal locus AMY2. The frequency of the AMY1/2 allele was calculated as 0.951; AMY3/2 and AMY4/2 seem to have a frequency of 0.001 in this sample.


Inheritance of PiM subtypes. A study of 151 families with a total of 242 children and of 142 mother-child pairs. Pi phenotypes were classified by isoelectric focusing in sera of 151 families with a total of 242 children and in sera of 142 mother-child pairs. The six common subtypes of PiM are genetically determined by three alleles named PiM1, PiM2, and PiM3. No exceptions to the postulated mode of inheritance have been found. The possibility of further heterogeneity of the intermediate variant PiM3 is discussed.


Lymphocytolytic activity of glucocorticoids in vitro: 51Cr release under equilibrium conditions. Cytolytic activity of glucocorticoids in vitro is assessed by measuring radiochromium release from steroid-treated thymic lymphocytes under the equilibrium conditions provided by a continuous-labeling technique. Isotope release is a glucocorticoid-specific effect produced at physiological concentrations and is virtually abolished by inhibitors of RNA and protein synthesis. The relative lytic potencies of the steroids tested are comparable to those reported for glucocorticoids as measured by other methods. This procedure not only possesses the advantages typical of isotopic techniques in general, but, in addition, circumvents the problem of "spontaneous" label release associated with the pulse-labeling method. It is a useful alternative to the morphologic examination of cells or the estimation of cell viability for determination of glucocorticoid cytolytic activity.


Differentiation of functionally active mouse T lymphocytes from functionally inactive bone marrow precursors. III. Induction of T-cell activities by growth of bone marrow on feeder layers prepared from mouse thymocytes. The ability of small and large T precursor cell pools to differentiate into mature functional T lymphocytes on thymus feeder cell layers or in the presence of a supernatant prepared from such feeder layers was compared. Evidence was obtained showing that the small cell pool differentiated as effectively in the presence of soluble factors prepared from the thymus cultures as in the presence of the thymus cells themselves. In contrast, large stem cells apparently needed cellular contact with the thymus feeder cultures in order to promote differentiation, though a time point could be found at which further differentiation from this precursor pool would also occur in the presence of the thymus culture supernatant only. These data concurred with evidence from in vivo experiments for two pools of T-cell precursors which differentiated in thymectomized or thymus-intact recipients respectively.


Studies on hyperlymphoid mice.

Studies on hyperlymphoid mice. Injection of syngeneic, but chromosomally distinguishable, lymph node or spleen cells into adult mice resulted in both T- and B-lymphocyte chimaerism. In spleen-injected mice haematopoietic chimaerism was also established. It appeared that the donor T lymphocytes were added to the host recirculating T-cell pool so that a hyperlymphoid state was produced. The percentage of donor T lymphocytes declined very slowly in normal mice, but remained stable in adult-thymectomized animals. There was no evidence of a homeostatic mechanism involving destruction of excess peripheral T lymphocytes or grossly affecting the flow of T lymphocytes from the thymus into the recirculating T-cell pool. A preliminary model of the T-lymphocyte system is proposed.


The effect of adult thymectomy upon helper function in Xenopus laevis, the South African clawed toad. Carrier-primed helper activity enhancing anti-hapten (TNP-) responses in young adult Xenopus laevis reached peak levels 2-4 days after a low dose challenge (0.005% RBC) of unconjugated carrier. It fell quickly to negligible levels by 10 days. Adult thymectomized (ATx) animals were consistently unable to generate substantial levels of hapten-specific antigen-binding spleen cell (ABC) activity. Since ATx does not severely reduce splenic ABC responses to the high dose (10% RBC) of carrier used for hapten challenge, the results suggested a role for Xenopus adult thymus in carrier-specific enhancement of an anti-hapten response.


T and B lymphocytes in pituitary dwarf Snell-Bagg mice. The lymphocyte composition of the thymus and spleen from weaned (4 month old) hypopituitary dwarf Snell-Bagg mice were compared to those of their phenotypically normal littermates and of hormone (somatotropic hormone plus thyroxine)-treated individuals. Detection of cells bearing receptors for peanut agglutinin, physical analysis and measurement of in vitro reactivities to phytohaemagglutinin and concanavae intra-thymic lymphocyte population of dwarf mice. Examination of spleen-cell suspensions demonstrated a slightly higher frequency of T lymphocytes (Thy 1-2+  alpha-Naphthyl esterase+, high electrophoretic mobility) and lower frequency of B lymphocytes (surface immunoglobulin+, low electrophoretic mobility) in dwarf mice than in control mice. The degree of splenocyte responsiveness to T- and B-cell mitogens, however was similar in the two mouse types. High mobility (T) splenic cells were found to exhibit a smaller modal volume in dwarf mice (110 micron3) than in control mice (122 micron3) but this difference was not corrected by hormone administration. More pronounced were the quantitative differences between the spleens of hormone-deficient and normal mice. Thus, when expressed as a function of body weight, the numbers of splenic T and B lymphocytes in untreated dwarf mice were about half the corresponding values in hormone-reconstituted or normal littermates. These data suggested that in adult life, developmental hormones exert little direct effect on the thymus lymphocytes but influence the size of the pool of both peripheral T and B lymphocytes.


Restricted heterogeneity of antibody synthesized by T-cell deprived mice. Mice which had been thymectomized and injected with anti-thymocyte serum to remove long-lived recirculating T cells, initially failed to produce haemagglutinating and haemolysing antibody after injection of sheep erythrocytes. After six fortnightly injections of heterologous erythrocytes, however, haemolysin titres in the T-cell deprived mice were comparable to those in similarly challenged but immunologically intact animals. Isoelectric focusing of these sera indicated that the anti-sheep erythrocyte antibody eventually synthesized by the T-cell deprived mice was less heterogeneous than antibodies found in the sera of control mice.


Acid alpha-naphthyl acetate esterase: presence of activity in bovine and human T and B lymphocytes. Non-specific acid alpha-naphthyl acetate esterase (ANAE) activity was demonstrated in a majority of bovine peripheral blood lymphocytes, confirming and extending the observations on murine and human lymphocytes made by previous workers. Simultaneous study of both ANAE activity and spontaneous erythrocyte (E) or erythrocyte-antibody-complement (EAC) rosetting capability of the same bovine lymphocytes showed directly that, while 64.2 +/- 4.6 (SEM) % of bovine lymphocytes capable of forming E rosettes were ANAE positive, 38.3 +/- 0.8% of those forming EAC rosettes were also ANAE positive. Similar studies of human peripheral blood lymphocytes showed also that, while 80.6 +/- 2.2% of the lymphocytes capable of forming E rosettes were ANAE positive, 44.1 +/- 2.6% of EAC forming lymphocytes were ANAE positive. Thus the presence of ANAE activity in a majority of T lymphocytes and a significant proportion of B lymphocytes of both human and bovine peripheral blood is indicated. Human and bovine lymphocytes from phytohaemagglutinin (PHA)-stimulated cultures demonstrated greatly enhanced intensity of ANAE activity.


Immunologically mediated intestinal mastocytosis in Nippostrongylus brasiliensis-infected rats. To investigate mechanisms of mast-cell proliferation, we have utilized infection of Lewis rats with the intestinal nematode, Nippostrongylus brasiliensis, which induces a pronounced intestinal mast-cell hyperplasia. Adoptive transfer of 2 x 10(8) immune mesenteric lymph node cells (IMLN), collected 14 days post infection with 3000 third stage larvae (L3), into rats concurrently given 3000 L3 hastened the expected intestinal mastocytosis by up to 4-5 days. IMLN exhibited this mastopoietic activity in the presence but not in the absence of concurrent infection. Normal mesenteric lymph node cells did not show similar mastopoietic activity. Intestinal mastocytosis was delayed by sub-lethal irradiation (400 rad) but IMLN reconstituted the mast-cell response of such animals. The mastopoietic activity could not be attributed to worm antigen as antigen administered intravenously had no significant effect on mastocytosis and furthermore, antigen could not be detected in mastopoietically active IMLN suspensions used as a possible antigen source in passive cutaneous anaphylaxis tests. Immune serum (14 days post primary infection with 3000 L3) also hastened mastocytosis in infected rats, whereas normal serum did not. The IMLN may be an enriched source of intestinal mast cell precursors and, in addition, may contain a cell type(s) which regulates the differentiation and proliferation of such precursors.


A comparison between LCM virus-specific secondary cytotoxic T lymphocytes generated by Con A and by the homologous antigen. An investigation was made of the properties of cytotoxic T cells induced by Con A and exposure to LCM virus-infected cells. As a basis for such studies, the optimal conditions for in vitro Con A stimulation of in vivo LCM virus-primed C3H mouse splenocytes were determined. The most potent cytotoxicity was obtained when responder cells were cultured in the presence of Con A in a concentration of 2 micrograms/ml for 3 days, but strong cytotoxicity was also measured on days 2 and 4. When stimulation was performed by the homologous antigen maximal response was seen on day 4 although marked cytotoxicity was also noted on day 3. Effector cells produced by the two different procedures showed equally high degrees of cytotoxicity against LCM virus-infected target cells, whereas they did not appear cytotoxic against non-infected targets. If LCM virus-immune mice were treated intravenously with 280 micrograms of Con A per animal, moderate cytotoxicity was demonstrable in splenocytes from these mice 1, 2 and 3 days after treatment. The in vitro generation of secondary cytotoxicity by Con A as well as by the homologous antigen was found to be totally dependent on DNA synthesis. The reactivated cells were investigated for in vivo anti-viral effect by measuring their ability to protect intracerebrally LCM virus-infected mice from a fatal outcome of this infection. LCM virus-primed splenocytes stimulated by the homologous antigen caused complete protection, while Con A-reactivated cells did not protect at all. Secondary cytotoxic cells stimulated by Con A and by LCM virus showed fairly similar in vitro characteristics, but fundamentally different in vivo qualities.


Immunoglobulin synthesis by thymus B cells. Rabbit lymphoid cells transferred to newborn recipients synthesized donor-type immunoglobulin. Elimination of donor T cells did not affect this synthesis of donor immunoglobulin, while elimination of B cells abolished or significantly decreased the synthesis. The synthetic capacity of B cells from thymus was thirty-four times greater than the synthetic capacities of B cells from spleen, fifty-five times greater than that of mesenteric lymph nodes and 180 times greater than that of appendix cells. Synthetic activity of spleen cells ceased before the tenth day after transfer, while thymus cells might continue to synthesize immunoglobulin for a longer time. This was shown by comparing half-lives of donor immunoglobulin in the recipients' sera. Increasing the number of injected spleen cells (from 2 x 10(6) to 120 x 10(6)), resulted in a corresponding increase in donor Ig synthesis. With thymus cells, donor immunoglobulin increased with cell numbers up to 2 x 10(7) cells, above this dose there was no further donor immunoglobulin increase in the recipients' serum.


Studies on B-cell memory. I. Generation and exhaustion of B-cell memory by thymus-dependent antigen in T-cell depleted mice. Mice depleted of T cells by adult thymectomy, X-irradiation and reconstitution with syngeneic bone marrow cells, either untreated or treated with anti-Thy-1 serum and complement, were immunized intensively with alum-precipitated bovine serum albumin (AP-BSA) along with or without bacterial lipopolysaccharide (LPS), but no significant anti-BSA antibody response was detected. Priming of the T-cell depleted mice, however, either by a single injection of AP-BSA plus LPS or by multiple injections of AP-BSA without LPS, resulted in the generation of immunological memory. A single injection of AP-BSA without LPS was ineffective. The memory required the aid of syngeneic T cells to be recalled by the challenge with AP-BSA plus LPS. On the other hand, multiple injections of AP-BSA plus LPS did not cause the generation of memory and the response of these mice to the challenge was lower than that of unprimed control mice. These results suggest that (1) the anti-BSA response is highly dependent on the helper function of T cells, (2) the degree of T-cell requirement for the memory generation is very low, and (3) priming with too much strong stimulation in the absence of functional T cells leads to the suppression or abortion of previously generated immunological memory.


Immunoregulation by covalent antigen-antibody complexes. II. Suppression of a T-cell independent anti-hapten response. Covalent antigen-antibody complexes containing the hapten fluorescein were found to suppress the thymus-independent response to that hapten. This suppression required the presence of an intact Fc fragment in the complex but could still be established in the complete absence of T cells.


One-stage stimulation of human T-lymphocyte colony-forming units (TL-CFU) in a micro agar culture in glass capillaries. A one-stage micro-culture assay in glass capillary tubes was developed for colony growth of human T lymphocytes. Colony growth by this method did not depend on the addition of growth factors, human serum and erythrocytes. Advantages of this method are its low consumption of materials and the fact that high numbers of cultures can be prepared and evaluated within a relatively short period of time. Compared to the two-stage method, the one-stage method requires a higher number of seeded cells and a reduced agar concentration, suggesting that cell interactions are required for human T-lymphocyte colony formation.


Ceforanide (BL-S786) in the treatment of community-acquired bacterial pneumonia. Ceforanide (BL-S 786) is a new long-acting parenteral cephalosporin which has the major pharmacologic advantage of requiring only twice a day dosage. We treated 28 adult patients with community-acquired bacterial pneumonia using doses of 500 or 1000 mg every 12 hours. Twenty-four of 28 infections were due to Streptococcus pneumoniae and/or Hemophilus influenzae, and all pathogens were susceptible in vitro to both cephalothin and ceforanide. Patients were treated for a mean of 7.5 days, and all showed a good clinical and radiographic response with no mortality. Of the 13 patients with H. influenzae, the organism could still be recovered during therapy in 9/12 and post therapy in 3/8. One clinical superinfection (sepsis due to Pseudomonas aeruginosa) occurred during therapy. Side effects with therapy included thrombocytosis (15), asymptomatic eosinophilia (5), and mild elevation of the serum transaminases (3). These studies suggest that ceforanide is a safe and effective agent for the treatment of adult patients with bacterial pneumonia due to S. pneumoniae; further experience in therapy of H. influenzae is needed because of frequent failure of ceforanide to eradicate this organism from the sputum.


Serum and sputum antibiotic levels after ampicillin, amoxycillin and bacampicillin chronic bronchitis patients. Serum and sputum concentrations of ampicillin or amoxycillin were measured in patients admitted to hospital for acute exacerbations of chronic bronchitis with purulent sputum. Mean peak serum levels of nearly 12 mg/l were found after 1600 mg bacampicillin (mean peak level in sputum 0.85 mg/l). The serum and sputum concentrations after 750 mg amoxycillin and 800 mg bacampicillin were comparable (mean peak serum levels approximately 9.5 mg/l, sputum concentrations 0.4 to 0.5 mg/l) although the drugs were not given in equimolar doses. Results after 1000 mg ampicillin by mouth were less satisfactory (mean peak serum level 7.8 mg/l) and only 0.25 mg/l was attained in the sputum. Minimum inhibitory concentrations of ampicillin and amoxycillin were measured for 177 Haemophilus influenzae strains. Most of the ampicillin MIC values were between 0.125 and 0.5 mg/l but more of the strains required 0.5 mg/l of amoxycillin. The amoxycillin MIC values were often one or two dilutions higher than those of ampicillin (p less than 0.001).


A clinical comparison of ampicillin, ampicillin esters (bacampicillin and pivampicillin) and amoxycillin in acute exacerbations of chronic bronchitis. The results of antibiotic therapy in 271 patients suffering from acute exacerbations of chronic bronchitis are presented. The effectiveness of the better absorbed ampicillin esters (pivampicillin and bacampicillin) is confirmed, but side-effects from the pivampicillin present problems whereas bacampicillin is excellently tolerated, even in twice daily doses of 1600 mg. Amoxycillin, if given in 750 mg doses three times daily by mouth, is also safe and effective against Haemophilus influenzae. However, if accurate MIC results are not available for both ampicillin and amoxycillin, the lesser degree of sensitivity to amoxycillin suggests that use of an ampicillin ester (such as bacampicillin) is to be preferred. Prophylactic use of antibiotics in chronic bronchitis patients does not seem logical to us.


Hereditary complement (C2) deficiency with discoid lupus erythematosus and idiopathic atrophoderma. A family with hereditary deficiency of the second component of complement was studied. Three siblings were homozygous for C2 deficiency and two of them had associated skin diseases. One sister presented with idiopathic atrophoderma and the other had clinical and pathological manifestations of discoid lupus erythematosus. This is the first description of an association between idiopathic atrophoderma and C2 deficient state.


[New fluorescence microscopic technic for the identification of Trichomonas vaginalis]. A new fluorescent dye for the identification of trichomonas vaginalis is described. Specimens under investigation were mixed with 4',6-diamidino-2-phenylindol (Dapi) and examined using white light and UV-light. The combined use of the two techniques allows to recognize the mobile non fluorescing and the dead intensive yellow fluorescing trichomonads. Devitalized epithelial cells, bacteria and leucocytes in contrast to living trichomonas are stained yellow in a few minutes. This is of diagnostic help.


Proteins contained in thymus factor X bind to the surface of rat thymocytes. By means of an immunocytochemical method, using an antiserum directed against Thymus Factor X (TFX) the presence of TFX-cross-reactive antigen on the surface of rat thymocytes was demonstrated. In the electron microscope, the product of the immunocytochemical reaction was localized as a fine granular and electron opaque material on the surface of some medium-size and small thymocytes. It was shown that the intensity and distribution pattern of the reaction product on the surface of the thymocytes varied markedly. Preincubation of thymocytes with TFX resulted in more intense labelling. The presence of TFX and cross-reactive native thymic antigens on the surface of rat thymocytes as revealed by anti-TFX serum, allows us to suppose that these thymic proteins are bound to surface of the cells by specific surface receptors.


The comparative beta-lactamase resistance and inhibitory activity of 1-oxa cephalosporin, cefoxitin and cefotaxime. The beta-lactamase stability and inhibitory activity of 1-oxa cephalosporin, (6R,7R)-7-[[carboxy(4-hydroxyphenyl)acetyl]amino]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-5-oxa-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, was investigated and compared to that of cefoxitin and cefotaxime. There was no detectable beta-lactamase hydrolysis of 1-oxa cephalosporin, cefotaxime and cefoxitin when incubated with beta-lactamases of plasmid or chromosomal origin which were primarily cephalosporinases or enzymes which hydrolyzed both penicillins and cephalosporins. The beta-lactamase inhibitory activity of 1-oxa cephalosporin was comparable to that of cefoxitin and cefotaxime. At equal molar concentration of substrate and inhibitor, cefoxitin, cefotaxime and 1-oxa cephalosporin effectively inhibited cephalosporinase hydrolysis of cephaloridine. Cefoxitin and cefotaxime were more effective inhibitors than the 1-oxa cephalosporin against a Providencia enzyme, whereas cefotaxime and 1-oxa cephalosporin were more effective inhibitors of a Citrobacter cephalosporinase.


Crosslinking by ligands to surface immunoglobulin triggers mobilization of intracellular 45Ca2+ in B lymphocytes. Detailed studies of steady-state ion fluxes in murine lymphocytes were used to examine for possible ionic changes generated by surface Ig, the antigen receptor of B lymphocytes. When bound by ligands, surface Ig triggered the mobilization and release of 45Ca2+ from the cell interior by a transmembrane process requiring crosslinking of the bound receptors. This ionic event was unique for two reasons: (a) it did not occur when other common lymphocyte surface macromolecules were bound with rabbit anti-lymphocyte antibodies; and (b) it was not accompanied by a general perturbation of lymphocyte ionic properties such as a change in 42K+ fluxes nor did it depend on the presence of extracellular ions. Capping of surface Ig shares the same time sequence, dose response, requirement for crosslinking, and lack of dependence on extracellular ions. These correlations suggest that mobilization of intracellular Ca2+ may represent an early ionic signal for the contractile activation of lymphocytes that generates capping of surface Ig.


Psychiatric sequelae to cardiotomy.

Psychiatric sequelae to cardiotomy. Eight of twenty-six subjects undergoing cardiotomy had a diagnosable psychiatric syndrome after surgery. No difference between psychiatric and non-psychiatric cases was noted with respect to demographic data, pre-operative medical psychiatric history, anesthesia time, serial laboratory studies, or environment. Post-operative psychiatric cases, however, did have significantly higher exposure to psychoactive drugs during the pre- and post-operative period.


Rapid induction of morphological changes in human carcinoma cells A-431 by epidermal growth factors. The morphological effects of epidermal growth factor (EGF) on human carcinoma cells A-431 have been examined by scanning electron microscopy. These flat polygonal cells normally exhibit only small membrane folds, but show extensive ruffling and extension of filopodia within 5 min of exposure to EGF at 37 degrees C. This ruffling activity is transient, subsiding within another 5--15 min, but several other changes in surface morphology follow. Within the first hour of exposure to the hormone, the cell surface becomes exceedingly smooth and the nuclei seem to protrude above the plane of the otherwise thin monolayer, giving the cells a "fried egg" appearance. Cells at the edges of colonies gradually retract from the substrate, leading to reorganization, by 12 h, of the monolayer into multilayered colonies. EGF thus induces both rapid and long-term alterations in the morphology of these epidermoid cells.


Rapid stimulation of pinocytosis in human carcinoma cells A-431 by epidermal growth factor. Horseradish peroxidase (HRP) uptake was used to measure fluid-phase pinocytosis in monolayers of human epithelioid carcinoma cells (A-431). Histochemistry confirmed that cell-associated HRP was restricted to intracellular vesicles. Biochemical methods showed that HRP uptake in control cultures was directly proportional to the duration of exposure. The addition of low concentrations of epidermal growth factor (EGF) to the incubation media produced a 10-fold increase in the initial rate of pinocytosis. The EGF effect was rapid (within 30 s) but transient; the rate of pinocytosis returned to control levels within 15 min. Metabolic inhibitors reduced the EGF-stimulated rate of pinocytosis by greater than 90%. A conjugate of EGF and ferritin (F:EGF) was used to simultaneously compare the intracellular locations of EGF and HRP. Much of F:EGF was internalized in approximately 100-nm vesicles, while most of the HRP was located in much larger vesicles (range 0.1--1.2 micrometer) which also contained F:EGF. The tumor-promoter 12-0-tetradecanoyl-phorbol-13-acetate, which shares several biological activities with EGF, was also effective in stimulating an increase in the rate of pinocytosis.


Binding, internalization, and degradation of epidermal growth factor by balb 3T3 and BP3T3 cells: relationship to cell density and the stimulation of cell proliferation. Epidermal growth factor (EGF) stimulates the growth of both benzo[a]pyrene-transformed Balb 3T3 cells (BP3T3) and untransformed Balb 3T3 cells. We describe here the binding, internalization, and degradation of [125I]-EGF by BP3T3 cells and 3T3 cells. Binding of [125I]-EGF reaches a maximum after 45 to 90 minutes incubation at 37 degrees C. In both BP3T3 and 3T3 cells the extent of EGF binding required to stimulate DNA synthesis is density dependent; sparse cultures require a 15-30% occupancy to elicit a maximal response whereas dense cultures require a 70-85% occupancy. At physiological concentrations the total binding of [125I]-EGF to 3T3 cells is higher than to BP3T3 cells, and this difference increases at higher cell densities. The rate of degradation of [125I]-EGF is directly proportional to the total [125I]-EGF binding in each cell type. This supports the hypothesis that one cause of the diminished serum requirement of BP3T3 cells is a reduced rate of utilization of serum growth factors.


Insulin effects on protein synthesis and secretion in primary cultures of amphibian hepatocytes. The objective of the present study was to determine the effects of insulin on amphibian hepatocytes in primary culture. Hepatocytes were isolated from adult bullfrogs by collagenase perfusion and maintained as monolayers in serum-free medium. Cells cultured in the continuous presence of insulin exhibited a relatively constant rate of protein secretion over the first four to five days, whereas controls showed an almost three-fold decrease over the same time period. The decline in secreted proteins was equally represented in most exported proteins, except that serum albumin secretion showed twice as much of a decrease relative to the other proteins. The maintenance of protein secretion by insulin was the result of its effect on protein synthesis. The rate of protein synthesis was measured by the incorporation of (3H)-leucine into protein using culture medium containing 0.5 mM leucine, a condition where the specific radioactivity of leucyl-tRNA was shown to be equal to that of (3H)-leucine in the medium. Cultures maintained with insulin for 60 hours synthesized protein at two to three times the rate found in non-insulin treated controls whose rate of protein synthesis was first detectably decreased after nine hours of culture in the insulin-free medium. Sedimentation profiles of polyribosomes from hepatocytes maintained for 60 hours without insulin showed proportionately fewer ribosomes in large polysomes and more in monosomes and free ribosomal subunits than ribosomes from cells cultured with insulin. This result suggests that the decrease in protein synthesis found in the absence of insulin is due to a defect in initiation. Insulin does not exert its effect by regulating cellular levels of ATP; no change in ATP content was found in cells maintained with or without insulin. The results show that insulin maintains high levels of protein synthesis and secretion in amphibian hepatocytes. The hepatocytes in monlayer culture provide a system to study the molecular mechanisms involved in the translational control of protein synthesis by insulin.


Polyene macrolide antibiotic cytotoxicity and membrane permeability alterations. II. Phenotypic expression in intraspecific and interspecific somatic cell hybrids. Cytotoxicity and membrane permeability alterations induced by the polyene macrolide antibiotics filipin (FIL) and pimaricin (PIM) have been compared in parental intraspecific and interspecific somatic cell hybrids. B82 (mouse) and B1 (hamster) cells were found to be more resistant than RAG (mouse) parental cells to both polyene macrolides as indicated by 24-hour survival, 72-hour viability, and growth rate. Analysis of both intraspecific and interspecific somatic cell hybrids indicated that polyene macrolide resistance was being expressed even in the presence of the polyene macrolide-sensitive (RAG) genome. Where one of the two parental cell types is relatively polyene macrolide resistant, the use of specific polyene macrolides may prove efficacious as half-selective agents in cell hybridization.


Presence of epidermal growth factor receptors and influence of epidermal growth factor on proliferation and aging in cultured smooth muscle cells. Epidermal growth factor (EGF) at nanomolar concentrations stimulated DNA synthesis in confluent, serum-starved cultures of calf aorta and human uterine smooth muscle cells. Stimulation of DNA synthesis in lens epithelial cells was studied for comparison. L and D-ascorbic acid potentiated the effect of serum and EGF on DNA synthesis in calf aorta cells. In contrast L-ascorbic acid had minimal potentiating effect with serum and no effect with EGF present along with serum on DNA synthesis in human uterine smooth muscle and rabbit lens epithelial cells. EGF and ascorbic acid increased cell number when added to stationary phase cultures. Specific binding of 125I-labelled EGF to smooth muscle cells was demonstrated. Receptor concentration in calf-aorta smooth muscle cells was higher in dense cultures compared to sparse cultures. The time course of binding and dissociation of 125I-labelled EGF was similar in "dense" and "sparse" cultures. Human uterine smooth muscle cells in culture exhibited a finite lifespan. There was no stimulation of DNA synthesis in response to serum and EGF in cells of high population doubling level (PDL); although 125I-labeled EGF binding was higher in old cells (high PDL) compared to young cells (low PDL). This increase in binding was shown to be due to changes in the concentration of receptors without changes in their affinity for EGF.


Properties and separation of T lymphocyte growth stimulatory activity (TL-GSA) and of granulocyte-macrophage colony stimulatory activity (GM-CSA) produced separately from two human T lymphocyte subpopulations. We have previously identified two stimulatory activities affecting blood cell maturation in PHA-stimulated human lymphocytes conditioned medium (PHA-LyCM). One was granulocyte-macrophage colony stimulatory activity (GM-CSA), and the other was T lymphocyte growth stimulatory activity (TL-GSA) in suspension culture. In this paper we have shown that although both activities can be produced from purified non-adherent human T lymphocytes, they are produced from two distinct subpopulations. The production of these activities was greatly enhanced by T cell mitogens. Both protein factors were relatively heat stable (56 degrees, 30 minutes), were sensitive to trypsin treatment and were specific for primate blood cells. These two activities were fractionated by means of ammonium sulfate precipitation, Sephadex G-150 gel filtration, DEAE cellulose and Con A-Sepharose column chromatographies. MW of the major peak estimated from the elution volume of gel filtration in the presence of 0.5 M NaCl was 40,000 for GM-CSA and 13,000 for TL-GSA. Results from Con A-Sepharose column showed that while about 70% of TL-GSA was bound to Con A, less than 25% of GM-CSA was bound. These observations show that the majority of TL-GSA and GM-CSA were separable by these two conventional column chromatographic methods.


Biotypes of Haemophilus encountered in clinical laboratories. The biochemical characteristics of 464 strains of Haemophilus influenzae and 83 strains of Haemophilus parainfluenzae isolated over an 18-month period are described. Of 22 characteristics obtained, only 6 were necessary to biochemically identify and biotype the isolates. The key substrates or tests were urease, ornithine, indole, o-nitrophenyl-beta-D-galactopyranoside, sucrose, and xylose. Five biotypes of H. influenzae and four of H. parainfluenzae were commonly recognized. Some strains were encountered which could not be accommodated in the recognized taxa but which constituted separate biotypes of the two species, H. influenzae biotype I was recovered principally from blood, cerebrospinal fluid, and upper respiratory secretion, and biotypes II and III were recovered from eye and sputum cultures. Biotype I was recovered primarily from children less than 1 year of age, whereas biotypes II and III were from persons 1 to 5 years old and from those over 20 years of age. Multiple isolates recovered from the same patient were almost always of the same biotype. Strains of H. parainfluenzae were isolated primarily from sputum, with others being isolated from body sources such as dental abscesses, gastric aspirates, and peritoneal fluid. An inverse relationship was noticed between hemolysis and mannose fermentation among H. parainfluenzae biotype III strains, whereas the relationship was absent among the other biotypes.


Sea blue histiocytosis in a patient with chronic non-neuropathic Niemann-Pick disease. A patient with Niemann-Pick disease is reported together with family studies. Her liver and bone marrow were shown to be infiltrated with sea blue histiocytes. Other organs, spleen and lung, were presumably also involved but histological proof was not obtained. Enzyme assay of leucocytes, lymphocytes, and cultured skin fibroblasts showed the patient to be deficient in sphingomyelinase activity. In fibroblasts, activity was 5% of normal while for the parents activity was about 50% of normal. The expected partial deficiency was not found using leucocytes or lymphocytes from the parents. Heat stability studies on fresh fibroblast extracts from the propositus indicated that residual sphingomyelinase activity was slightly more labile than that of the controls. It seems clear that chronic Niemann-Pick disease without neurological involvement is associated with sea blue histiocytosis.


T-cell nature of leukaemic cells in a case of Sézary's syndrome with 'null-cell' features. alpha-naphthyl acetate esterase (ANAE) activity has been investigated in leukaemic cells from peripheral blood in a typical small-cell Sézary syndrome (SS) case in which cerebriform mononuclear cells failed to form E rosettes. The 'dot-like' ANAE positivity found in the majority of these neoplastic cells strongly supports a T-cell origin. In addition, a non-monocytic, non-B-cell nature of Sézary cells is indicated by the lack of Ia-like antigens. Finally, there is evidence of a distinct portion of Sézary cells simultaneously expressing ANAE activity and Fc IgM receptors.


Use and interpretation of Schlichter's test on Haemophilus influenzae: relation of in vitro to in vivo results for cefamandole. When Haemophilus influenzae infections are treated by an antibiotic acting on the bacterial wall, the adequacy of antimicrobial therapy can be assessed by Schlichter's test. This test may be carried out using Mueller Hinton broth (or Mueller Hinton broth with 50% pooled serum and a supplement of Ca++ and Mg++) supplemented with Fildes' enrichment and an inoculum adjusted to the 0.5 McFarland turbidity standard diluted 200x. However, correct reading of end points can be obtained only by phase contrast microscopic examination, which allows the establishment of good correlation between the in vitro and in vivo findings. In patients with lung infections successfully treated with cefamandole, the presence of spheroplasts in samples derived from Schlichter's tests correlates well with clinical improvement and eradication of the pathogenic organism checked by transtracheal aspiration.


Cell-mediated immunity in Darier's disease. Darier's disease, often complicated by skin infections, has been linked with a depressed cell-mediated immunity. Four moderate to severely affected patients with a negative history of skin infections were studied. T cell function was evaluated by standard delayed hypersensitivity skin tests, enumeration of peripheral T cell-mediated immunity was found. The significance of the results when compared with prior studies is discussed.


Electrocardiographic gating in positron emission computed tomography. Electrocardiographic (ECG) synchronized multiple gated data acquisition was employed with positron emission computed tomography (ECT) to obtain images of myocardial blood pool and myocardium. The feasibility and requirements of multiple gated data acquisition in positron ECT were investigated for 13NH3, (18F)-2-fluoro-2-D-deoxyglucose, and (11C)-carboxyhemoglobin. Examples are shown in which image detail is enhanced and image interpretation is facilitated when ECG gating is employed in the data collection. Analysis of count rate data from a series of volunteers indicates that multiple, statistically adequate images can be obtained under a multiple gated data collection format without an increase in administered dose.


Quantitation in positron emission computed tomography: 2. Effects of inaccurate attenuation correction. Effects of inaccurate attenuation correction on quantitation in positron emission computed tomography were investigated by both computer simulation and theoretical analysis, followed by experimental verification with phantom measurements. It was found that, due to the finite width of detector responses, compensation of scan measurements by measured attenuation correction factors does not completely correct for attenuation. For FWHM greater than 1.8 cm, errors in object size and in activity levels of reconstructed images are more than 4%. Mismatches between true and assumed attenuation media are found to give large errors. These mismatches include object shift, incorrect size and shape, uniform and nonuniform mismatches in attenuation coefficients, and nonzero background levels. Errors as large as 14% in the reconstructed activity level can be introduced by a 5 mm shift in object position. A 5% error in the cylinder diameter can give differences between the total reconstructed activity and true value more than 12%. Strong image distortions, which could lead to incorrect interpretations of images are created by object shift and by nonuniform mismatches in attenuation coefficient. It was also found that statistical noise in measured attenuation correction factors can significantly increase noise levels in images, resulting in lower effective total counts for the emission scan. Effective total counts can be estimated as Neff = kNxmNem/(Nem + kNxm), where Nem and Nxm are total counts in emission and transmission scans, and k is a scanner dependent factor, which accounts for configuration differences.


Differentiation of retroperitoneal tumor using positron emission computed tomography. Positron emission computed tomography (ECT) with 11C-DL-tryptophan and 11C-aminocyclobutanecarboxylic acid (11C-ACBC) has produced some promising results in patients with retroperitoneal masses. This brief report gives an example of one patient in whom a retroperitoneal mass could not be differentiated from the pancreas by transmission computed tomography but was suspected to be of pancreatic origin based on arteriographic findings. The ECT scans with 11C-DL-tryptophan showed a normal pancreas, whereas the tumor localizing agent 11C-ACBC concentrated in a tumor surrounding the tail and body of the pancreas. This approach with dual radiopharmaceuticals for the differentiation of metabolically active neoplasms and normal tissues warrants further investigation.


Rosetting of human T lymphocytes with goat red blood cells: effect of treatment with 2-aminoethylisothiouronium bromide (AET) and comparison with AET treated sheep red blood cells. In vitro treatment of goat red blood cells (GRBCs) with the sulphydryl compound 2-aminoethylisothiouronium bromide (AET) increases their specific reactivity with human T lymphocytes without affecting the specificity of the reaction. AET-GRBCs bind to only part of T lymphocytes rosetting with AET-sheep red blood cells (SRBCs): the receptors for both types of RBCs are very simular if not identical, but display higher affinity for AET-SRBCs than for AET-GRBCs. Rosetting of T lymphocytes with AET-GRBCs may be useful to enumerate T lymphocyte subsets in patients with abnormality of the immune system and to fractionate T lymphocyte subpopulations.


Lymphocyte membrane alterations caused by nylon wool column separation. Use of the nylon wool column method to separate lymphocytes results in preparations which, while fully viable, have grossly depleted cellular levels of phospholipid, cholesterol and sialic acid. These membrane effects, which relate mainly to T cells, indicate that the method is unsuitable for use in biochemical analysis of lymphocytes.


Mixed agglutination in microtiter U-trays. The principle of mixed agglutination with cell cultures was applied to those cells which could not be obtained as monolayer cell cultures. Artificial monolayers of freshly isolated cells from various tissues were prepared in poly-L-lysine-coated wells of a microtiter U-tray. By means of this technique, murine alloantigens, H-2 private specificities and Thy-1 antigens were demonstrated on cells from thymus, brain, spleen, kidney and liver. Ia.2 antigen was detected on splenocytes but not on thymocytes or fibroblast L cells. Evidence was also presented that HLA A and B locus antigens could be demonstrated on human thymocytes. Sensitivity of this test was shown to be at least 100 times higher than that of the standard lymphocytotoxicity test for detection of HLA antigens.


Ineffectiveness of aprotinin on psoralen-UVA-(PUVA)-induced erythema. Because bradykinin constitutes a possible candidate for mediation of topical 8-methoxypsoralen-UVA-(PUVA)-induced erythema, aprotinin (Trasylol), inhibitor of kallikrein and interrupter of the cascade leading to kinin production was assessed in guinea pigs. Response was assessed at 24, 48, and 72 hr after topical PUVA and there was no significant difference between normal saline and aprotinin by intradermal or intraperitoneal routes of administration. The results of this study indicate that intradermal and intraperiotoneal aprotinin, in the dose and method tested, is not capable of significantly decreasing erythema induced by topical PUVA in guinea pigs.


Karyotype studies of cutaneous T cell lymphoma: evidence for clonal origin. Neoplastic lymphocytes from 3 patients with widespread cutaneous T cell lymphoma were karyotyped, using a chromosome banding technique. None of the patients had previously received chemotherapy. Morphologically homogeneous populations of abnormal T cells were available from 2 distinct body regions of 2 of the individuals and from the bone marrow of the third. Karyotypes from each of the 3 patients indicated monoclonality of their lymphoma. These observations suggest that extracutaneous dissemination of cutaneous T cell lymphoma involves spread of neoplastic cells derived from a single clone. If future studies can demonstrate that those neoplastic T cells actually localized in the skin are also progeny of a single malignant cell, a widely accepted concept that cutaneous T cell lymphoma is of multifocal origin will have to be reexamined.


Fibrin deposition and clearance in chronic granulomatous inflammation: correlation with T-cell function and proteinase inhibitor activity in tissue. Patterns of fibrin deposition were investigated by immunofluorescence microscopy in livers of thymus intact (TI) and athymic (AT) mice infected with Schistosoma mansoni. Thrombin and fibrinolysis inhibitor activity in tissue extracts also were measured. In TI mice fibrin was detected perivascularly by 6 weeks after infection and at 8 weeks it was found over the granulomas as they developed. Fibrin was cleared from the center of granulomas by 10 weeks. Thrombin inhibitor activity increased at 4 to 6 weeks but declined below control levels later as granulomas formed. Fibronolysis inhibitor activity, on the other hand, peaked at 9 to 12 weeks after infection. In AT mice extensive fibrin deposition was detected in the liver throughout the period when smaller and incomplete granulomas developed. Central clearing did not occur. Thrombin inhibitor activity greatly increased by 8 weeks after infection but fibrinolysis inhibitor activity remained unchanged. These findings suggest that fibrin deposition and firbinolysis are orderly events regulated in the lesions by proteinases and their inhibitors and this seems to be a general tissue reaction in the early stage of chronic granuloma formation. Since local clearance of fibrin in vivo and fibrinolysis inhibitor activity from tissue extracts studied in vitro are more evident in TI mice than in AT mice, it appears that T cell fu;ction is important in modulating the tissue response during granulomatous inflammation.


[Idiopathic scoliosis and equilibrium disturbance (author's transl)]. The body balance is maintained by the complexed nervous system in the brain stem and the cerebellum. However, the amount of neurological information obtained from this area is relatively small, so that a fine abnormality is easily overlooked or could not be definitely elicited. Examination of the ocular movement becomes often a powerful tool to detect an indiscrete lesion in the brain stem and the cerebellum which otherwise fails to be demonstrated neurologically. With a hypothesis that equilibrium disturbance might be playing a role in the development of idiopathic scoliosis, extensive neuro-otological studies were carried out in three groups of individuals: 169 cases of idiopathic scoliosis, 19 cases of congenital scoliosis and 488 young girls with no scoliosis. The examination included Mann's test, deviation tests and other neuro-otological tests with the special reference to the observation of the ocular movement. 1. There was no case in the entire groups which presented positive Mann's test or deviation abnormality. 2. With the aid of electronystagmography (ENG), the presence of nystagmus was detected in 71 out of 169 cases (42.0%) with idiopathic scoliosis, 9 out of 19 cases (46.7%) with congenital scoliosis and 5 out of 488 girls (1.0%) with no scoliosis. This fact was highly suggestive of the possibility of equilibrium disturbance in the background of scoliosis. 3. However, even with positive ENG findings, few cases showed abnormality on optokinetic nystagmus (OKN) test or eye tracking test, so that definite localization of the lesion was impossible in the majority of the cases with the abnormal ocular movement. 4. There was no significant correlation between the rate of the abnormal ocular movement and the age of the patient, the magnitude of the spinal curve or the curve pattern. The co-existence of abnormal ocular movement and scoliosis was proved to be statistically significant, suggesting that equilibrium disturbance might be playing a role in the multifactorial etiology of idiopathic scoliosis, although the real pathomechanism still remains to be studied.


A new nystagmographic test for the elicitation of sub-clinical vertebro-basilar insufficiency and poor cochleo-vestibular vascularization. The nystagmus due to vertebro-basilar artery insufficiency is that nystagmus which is induced by torsion and extension of the neck, continued for more than 3 minutes, after nystagmus of other types has been excluded. It is of vascular origin, in contra-distinction to nystagmus of cervical origin, which is proprioceptive. It permits the detection of sub-clinical vertebro-basilar insufficiency, and it should be elicited before any operation which requires prolonged cervical rotation. It is a very simple method which can be performed at the end of the classical torsion swing test, and we have found the results very informative.


Cardiac pacemaker oscillation and its modulation by autonomic transmitters. 1. A scheme is presented which summarizes the activation and deactivation of the membrane currents which underlie pacemaking in the natural pacemaker of the heart. 2. Experimental evidence (mostly obtained using the voltage-clamp technique) for the properties of the time-dependent membrane currents in pacemaking tissue of the frog and the rabbit is discussed. 3. The mode of the inhibitory action of acetylcholine on pacemaker cells is considered. In the amphibian pacemaker cell, acetylcholine probably reduces slow inward current (as it certainly does in amphibian atrium) but in mammalian sino-atrial node it seems that such action, if present at all, is much less marked. In the pacemakers of both amphibian and mammal, acetylcholine greatly increases outward potassium current and there is recent evidence that it may do so by opening up a special acetylcholine-activated potassium channel. 4. Adrenaline greatly increases the slow inward current in pacemaker as in other cardiac tissues. This increase, together with (in mammal at least) an increased change of an additional pacemaking current, overrides an adrenaline-induced increase in outward current and leads to acceleration of the pacemaking rate. 5. The Appendix contains a brief consideration of the experimental and theoretical basis for the method of exponential separation of outward current components in the presence of the extracellular potassium accumulation that inevitably accompanies the flow of outward membrane current.


T cells recognize minor histocompatibility antigens on H-2 allogeneic cells. B10.A animals were rendered tolerant to B10.M spleen cells by injection of (B10.A X B10.M)F1 cells into neonates. Adult animals accepted B10.M skin grafts and failed to generate cytotoxic effector cells in vitro against B10.M H-2 antigens. In vivo inoculation of tolerant animals with A.CA spleen cells, followed by in vitro challenge with similar cells, resulted in the generation of cytotoxic effector cells that had specificity for the A strain minor histocompatibility (H)-antigens in the context of the H-2f haplotype. If these animals were boosted in vitro with A strain spleen cells, cross-priming could be demonstrated, whereby the cytotoxic effect was restricted by the H-2a haplotype. These data indicate that at least two sets of T cells co-exist in tolerant animals, one capable of recognizing antigens in the context of the host H-2 haplotype, and the other able to recognize antigens in the context of the tolerated H-2-allogeneic haplotype. Because tolerant animals inoculated with A-strain spleen cells in vivo and boosted in vitro with A.CA spleen cells failed to generate a cytotoxic effect against A.CA, it is unlikely that minor H-antigens need to be processed by host lymphoreticular cells.


Synthesis and processing of molecules bearing thymus leukemia antigen. Thymus-leukemia (TL) antigens are expressed in murine lymphocytes under strict developmental regulation. To elucidate the molecular basis of TL expression, we have identified the molecular species that react with TL antiserum. At least three species can be resolved by metabolic radiolabeling of thymocytes and ASL1 leukemia cells, lysis, immune precipitation, and sodium dodecyl sulfate-polyacrylamide. After a brief incubation with [35S]methionine, the only radioactive molecule recognized by TL antiserum is a homogeneous species with an apparent Mr of 45,000 daltons. This molecule, 45K TL, includes high-mannose-type carbohydrate attached to a 45,000 dalton glycosidase-resistant backbone. In this form, 45K, it is never exposed on the cell surface. If pulse-labeled cells are further incubated with nonradioactive methionine before lysis, however, radioactivity disappears from the 45K TL species and appears in the slower migrating species 46K and 48K TL. Thus, 46K and 48K appear to represent products generated from the 45K TL precursor by posttranslational modification. These TL forms are displayed on the cell surface; they lack high-mannose carbohydrate but evidently include acidic complex-type carbohydrate. Normal thymocytes from Qa:Tla-negative mice lack not only the surface forms of TL but also the intracellular 45K TL form. Peripheral lymphoid cells of Qa:Tla-positive mice synthesize none of these TL species. But the TL antiserum, which contains Qa antibody, recognizes a distinct gene product in spleen and thymus of Qa-Tla-positive mice. In its pulse-labeled form, this molecule, which may represent Qa-1, has an apparent Mr of 44,000 daltons, and consists of a glycosidase-resistant polypeptide core of only 35,000 daltons linked to more high mannose carbohydrate than 45K TL.


Production of auto-anti-idiotypic antibody during the normal immune response to TNP-ficoll. III. Absence in nu/nu mice: evidence for T-cell dependence of the anti-idiotypic-antibody response. Although athymic mice make an excellent immune response to the thymus-independent antigen trinitrophenyl-lys-Ficoll (TNP-F), nude mice of AKR/J and BALB/c strains lack the anti-idiotypic response that occurs in euthymic mice of both of these strains within the first 1--2 wk after injection of more TNP-F. Anti-idiotypic antibody-blocked (hapten-augmentable) anti-TNP splenic plaque-forming cells (PFC) do not occur at any time and serum anti-idiotypic antibody is absent in both congenitally athymic mice, and thymectomized, irradiated, bone marrow-reconstituted mice. Nevertheless, nu/nu mice do have PFC which can be inhibited by exposure to anti-idiotypic antibody produced in +/+ mice. As a consequence of the failure to produce anti-idiotypic antibodies, the anti-TNP PFC response is athymic as compared to euthymic mice is of greater magnitude, declines less precipitously, and shows an increase rather than a decrease in affinity between days 4 and 7 after antigen injection. It is concluded that the anti-idiotypic antibody response is thymus dependent and that athymic mice lack a helper cell required for the induction of anti-idiotypic antibodies.


Biochemical and biological characterization of lymphocyte regulatory molecules. I. Purification of a class of murine lymphokines. Murine spleen cells activated by concanavalin A (Con A) in culture produce a class of lymphokine molecules which possess biological activity in a number of lymphocyte response assays. Lymphokines with a mol wt of 30,000, as estimated from gel filtration studies, can be resolved into two components which differ by charge, with isoelectric point (pI) values of 4.3 and 4.9, respectively. Both components stimulate (a) the growth of established T-cell lines in culture, (b) the proliferation of thymocytes in the presence of Con A under culture conditions where Con A alone is nonmitogenic, (c) the induction of antibody responses to heterologous erythrocyte antigens in athymic (nude) spleen cultures, (d) the generation of cytotoxic T lymphocytes (CTL) in thymocyte cultures, and (e) the generation of CTL in nude spleen cultures. In each of these culture systems we suggest that the assays are detecting a single class of lymphokine which acts directly on activated T cells. Nonactivated T cells must be stimulated by either antigen or mitogen before becoming responsive to lymphokine, but do not require antigen or mitogen for continued growth with lymphokine. The two molecular species, separable by isoelectric focusing are referred to as the T-cell growth factor (TCGF). A lymphokine, similar in size (30,000 daltons) to TCGF but heterogeneous in charge (pI 3.0--4.0), stimulates immune responses to erythrocyte antigens in T-cell-depleted spleen cultures but has no stimulatory activity in the other lymphocyte assay systems described. The data have been interpreted as showing the two molecular forms of murine TCGF (pI 4.3 and 4.9) are responsible for many of the lymphokine activities described elsewhere as thymocyte mitogenic factor, nonspecific T-cell-replacing factor and killer helper factor or costimulator. The other lymphokine, separable from TCGF by charge, appears to have true T-cell-replacing activity.


Delayed-type hypersensitivity to allogeneic cells in mice. III. Sensitivity to cell-surface antigens coded by the major histocompatibility complex and by other genes. DTH could be induced to cell-surface antigens coded by either H-2 or non-H-2 genes. Sensitivity was more readily induced across I region than across K- or D-region differences. The presence of an I-region difference during sensitization did not significantly increase the DTH response to K- or D-region-coded antigens. Macrophage processing appeared to be the major route of sensitization to background antigens. Thus, high levels of sensitivity were achieved equally well using viable or disrupted cells, the response was independent of the H-2 haplotype of the allogeneic cells, and transfer was restricted to the K end of the host H-2 complex. Although sensitization to H-2 antigens was obtained with disrupted cells, transfer of sensitivity against viable cells was unrestricted. This suggests a minor role for macrophage processing in sensitization to H-2 antigens.


Ontogeny of B cells in CBA/N mice. Evidence for a stage of responsiveness to thymus-independent antigens during development. This paper deals with the CBA/N mice, a strain bearing a genetic defect in their B-cell compartment. By using a previously described system we have been able to show that the immature cells of CBA/N mice are functionally indistinguishable from normal immature cells, in that both can be triggered to respond to thymus-independent (TI) antigens, provided they are supplied with helper T cells. When the maturation is completed, CBA/N B cells are unable to respond to TI antigens (like lipopolysaccharide and polyvinyl pyrrolidine) irrespective of the presence of helper T cells, whereas normal mature B cells have grown able to respond without any help. These data allow us to reject the hypothesis that CBA/N mice are arrested at an immature stage and clearly support the idea that they have deviated during development so that only thymus-dependent B cells develop.


The purified protein derivative of turberculin, a B-cell mitogen that distinguishes in its action resting, small B cells from activated B-cell blasts. The purified protein derivative of tuberculin (PPD tuberculin) stimulates approximately one of two lipopolysaccharide (LPS)-activated B-cell blasts of C57BL/6J nu/nu spleen cells to continued clonal growth and maturation to IgM and IgG secretion. It alwo stimulates background, in vivo-activated large cells of normal C57BL/6J nu/nu spleen to growth and Ig secretion, at a frequency of approximately 1 of 100 large spleen cells. PPD tuberculin, therefore, is a polyclonal B-cell activator for B-cell blasts. Many single murine splenic B cells (approximately 50%) appear to have reactivities, and therefore probably receptors, for LPS and PPD tuberculin. PPD tuberculin does not stimulate small, resting B cells to growth as measured by the number of cells in culture and by thymidine uptake. However, it stimulates approximately one-fourth of all spleen cells to blast transformation. The large-size blast cells secrete IgM and, therefore, form plaques in the protein A plaque assay. IgG-secreting, plaque-forming cells develop at later stages of stimulation, indicating that the switch from IgM to IgG may occur without division in single, stimulated B cells. Stimulation of resting B cells to maturation by PPD tuberculin is polyclonal. Thus, approximately 1 in 10(2) IgM-secreting plaque-forming cells form plaques with trinitrophenyl-substituted sheep erythrocytes, 1 in 450 do so with horse erythrocytes, and 1 in 10(3) with sheep erythrocytes. Furthermore, the number of Ig-secreting cells developing from small, resting cells without growth in cultures with or without filler thymus cells suggests polyclonal activation by PPD tuberculin to maturation only of at least one out of four small, splenic B cells.


Role of prethymic cells in acquisition of self-tolerance. The sequential character of T-lymphocyte development as it pertains to the stage at which self-tolerance is acquired was investigated. Three phases were studied, defined here as prethymic, intrathymic, and postthymic as determined by the timing of thymus implantation. The model utilized was the temporal pattern of skin graft rejection in thymusless BALB/c nude mice implanted with allogeneic, C57BL/6J, or syngeneic thymuses before or after skin grafting; in some instances, F(1) hybrid spleen cells were also given to newborns or young adults. These experiments in nude mice showed that, (a) self-tolerance could be established despite the absence of the host's own haplotype in the implanted thymus; (b) recently emigrated postthymic cells could already discriminate self from non-self; (c) specific neonatal tolerance could be induced in nudes by inoculation of F(1) hybrid cells; (d) nudes showed a higher capacity for induction of neonatal tolerance than did normal littermates. These findings indicate that the process of self-tolerance in the T cell's lineage begins during the prethymic state early in ontogeny.


A cell-mediated reaction against glomerular-bound immune complexes. Lewis rats were given a single i.v. injection of soluble immune complexes containing human serum albumin (HSA) and rabbit anti-HSA antibodies, prepared in antigen excess. This resulted in localization of HSA and rabbit gamma globulin (RGG) in glomerular mesangial regions without producing definite histologic changes. 24 h after the injection of immune complexes, groups of these rats received lymph node cells or T-cell preparations from syngeneic donors sensitized to RGG, HSA, or ovalbumin; another group received no cells. All of these groups and a group of normal control rats were given injections of [3H]thymidine at 18, 27, and 44 h. The animals were killed 48 h after the time of cell transfer. In histologic sections, glomerular abnormalities were found only in some of the animals that had received immune complexes and lymph node cells or T-cell populations from donors sensitized to HSA or RGG; the lesions were characterized by focal and segmental increase in cells in mesangial regions. Autoradiographs revealed significantly greater numbers of labeled cells in mesangial regions and glomerular capillaries in the groups that had received immune complexes and cells from HSA- or RGG-sensitized donors than in any of the other groups. Electronmicroscopic studies suggested that the increase in cellularity in mesangial regions resulted from an influx of mononuclear phagocytes. The findings indicate that cell-mediated reactions can be initiated by the interaction between sensitized T lymphocytes and antigens present in immune complexes within mesangial regions.


Soluble factors in tolerance and contact sensitivity to 2,4-dinitrofluorobenzene in mice. III. Histocompatibility antigens associated with the hapten dinitrophenol serve as target molecules on 2,4-dinitrofluorobenzene-immune T cells for soluble suppressor factor. Previous studies have shown that suppression of 2,4-dinitrofluorobenzene (DNFB) contact sensitivity by soluble suppressor factor (SSF) requires that the donor of immune lymph node (LN) cells and of SSF share either the H-2K and/or H-2D region of the major histocompatibility complex. Thus, target or acceptor molecules for SSF appear to be coded for by genes within the H-2K and H-2D loci. Experiments were done to investigate the nature of these target molecules and to determine what cell types expressed them. It was found that purified lymph node T cells are suppressed by SSF indicating that T cells express the acceptor molecules. Adsorption experiments showed that the only cells capable of adsorbing the suppressor factor are DNFB-immune T cells from donors which share with the factor-producing strain either the H-2K or H-2D locus. This adsorption can be specifically blocked by pretreating the immune LN cells with antibodies directed against H-2K and/or H-2D determinants or against the hapten DNP but not by antibodies against Ia or theta-antigens. Collectively, these results indicate that the target molecules are expressed only by DNFB-immune T cells and are comprised of histocompatibility antigens associated with DNP.


B-cell differentiation in the CBA/N mouse. I. Slower maturation of mitogen and antigen-responsive B cells in mice expressing an X-linked defect. The effect of age on the mitogenic and antigenic responsiveness of B cells is examined in spleen cell cultures of CBA/N and (CBA/N X DBA/2) F1 mice. Spleen cells from young male F1 mice (4- to 6-wk old) show lower mitogenic responses to lipopolysaccharide, a lower frequency of sheep erythrocytes (SRBC)-reactive B-cell precursors, and a lower percentage of Ig-bearing cells than age-matched female F1 mice. The expression of all three functions were found to increase with the age of the F1 male mice. Whereas male F1 mice at 60 wk of age showed an equivalent percentage of Ig-bearing spleen cells and a similar mitogenic responsiveness to LPS when compared to adult female F1 mice, the frequency of SRBC-reactive B-cell precursors remained threefold lower. These findings reveal that there is a slower maturation of B cells in mice expressing the X-linked defect and suggests that the defect has differential effects on the mechanisms of antigen and mitogen activation of B cells.


Influence of the sex-linked defect in CBA/N mice on autoimmune responses to isologous erythrocytes. Ability to overcome the defect with age. Normal mice spontaneously develop plaque-forming cells (PFC) specific for antigens on modified self erythrocytes (bromelain-treated mouse erythrocytes [BrMRBC] antigens). Our study demonstrates that the sex-linked defect that results in the inability of CBA/N mice to respond to several T-independent antigens (TI-2 antigens) also regulates the autoantibody response to BrMRBC antigens. Thus, in CBA/N homozygous mice and male F1 offspring of CBA/N-mothered crosses, e.g., (CBA/N X NZB)F1 males, such PFC are absent. To examine whether specific autoreactive B cells are present in defective mice, the latter were stimulated either nonspecifically with the mitogen LPS or by infection with lethal malaria (17XL Plasmodium yoelii) known to induce anti-BrMRBC PFC specifically. The results indicate that modest antibody responses to self antigens could be induced in young (5- to 7-wk old) defective mice and that these responses increased as a function of age. The data is consistent with the view that the defect in CBA/N mice does not result from an absence of functional anti-BrMRBC B cells but rather from low frequencies of the specific precursors, which can be triggered and expanded with age probably by environmental stimulations.


Cation selectivity of acetylcholine-activated ionic channel of frog endplate. Ionic selectivity of the acetylcholine-activated ionic channel of frog endplate membranes to various organic cations has been studied. The ratio of test cation permeability (PX) to sodium permeability (PNa) was estimated by two methods, one based on the measurements in test cation solutions of the amplitude of transient depolarization induced by iontophoretic application of acetylcholine, and the other on the measurements of the reversal potential for the membrane current induced by iontophoretic application of acetylcholine under voltage-clamp conditions. The endplate channel is relatively nonselective to various test cations. The permeabilities relative to Na are ammonium (1.71), formamidine (1.49), methylamine (1.39), hydrazine (1.35), and Li (0.76), as measured from the reversal potential for acetylcholine currents, and guanidine (0.74), aminoguanidine (0.20), methylguanidine (0), and choline (0) as measured from the amplitude of acetylcholine potential. Methylguanidine and aminoguanidine block the endplate channel with the apparent dissociation constants of 0.5 and 15 mM, respectively. Based on these data, the dimensions of selectivity filter of acetylcholine-activated channel appear to be slightly larger than those of the sodium channel of frog nodes and smaller than those of the epithelial membrane of gallbladder of frogs and rabbits.


Simplified media for the growth of Haemophilus influenzae from clinical and normal flora sources. The nutritional requirements of 43 strains of Haemophilus influenzae isolated from clinical and normal flora sources were investigated. Two defined minimal media were developed by modifying the medium of Herriott et al. (1970):74% of the strains could grow on the minimal media and Herriott's medium; the remaining strains could not grow on any of these media.


Clinical signs of visual-vestibular interaction. Visual suppression of vestibulo-ocular reflexes (induced vestibular nystagmus) differs between normal subjects and patients with various neurological disorders. Abnormalities of VOR suppression were associated consistently with abnormalities of the visual and/or vesitibular oculomotor reflexes and in particular with abnormal smooth pursuit eye movements in the direction of abnormal suppression. Absent VOR suppression in a gaze position of, and in the same direction as, a spontaneous nystagmus was found exclusively in patients with spontaneous nystagmus of central nervous system origin. Conversely, ability to suppress in these circumstances was found only in patients with spontaneous nystagmus of peripheral labyrinthine origin. Suppression of VOR was abnormal ipsilaterally in patients with unilateral cerebral hemisphere lesions and abnormal in both the horizontal and vertical planes in patients with basal ganglia lesions. Failure of VOR suppression in the absence of spontaneous nystagmus indicates a supratentorial lesion.


Latent herpes simplex virus trigeminal ganglionic infection in mice and demyelination in the central nervous system. Mice were inoculated with herpes simplex virus (HSV) type 1 by gently scraping the skin of the nose with a fine needle. About 80% of the animals developed latent inapparent HSV infections in trigeminal ganglia. Virus was demonstrable for at least 6 months post inoculation (p.i.) by cocultivation of ganglionic tissue with GMK cells. Histologically, trigeminal ganglia revealed infiltrations of inflammatory cells even 6 months p.i. In addition, lesions occurred in the brainstem corresponding to the entry of trigeminal roots, trigeminal tracts and nuclei. Inflammatory cell infiltration, disruption of myelin sheaths and macrophages laden with myelin degradation products were observed 7 days p.i. Fourteen to 30 days p.i. electron microscopy demonstrated completely naked axons. In the transitional region of the trigeminal root denuded axons occurred in the central part of the region while the peripheral myelin, bordering the demyelinated central segments, was intact. Small areas of demyelination were still detectable 3 and 6 months p.i. but there were then also signs of remyelination. Possible mechanisms causing the demyelinations are discussed.


Studies on the humoral and cell-mediated immune response in a patient with Mollaret's meningitis. A patient with Mollaret's meningitis with protracted course and a strong immune response within the central nervous system is described. The findings of IgG bands in agarose gel electrophoresis of CSF together with elevated CSF IgG index values are in accordance with intrathecal synthesis of IgG, possibly against an antigen persistent in the CNS. Subgrouping of CSF lymphocytes morphologically (B/T, active T cells) and functionally (stimulation with mitogens) showed that these cells differed from peripheral blood lymphocytes. A defect in the regulatory function of T cells may be responsible for the continuing pathologic process in Mollaret's meningitis.


The importance of maintenance in implementing change: an experience with problem-oriented recording. Most change implementation efforts focus on the initial acquisition-of-skill phase with little attention to maintaining skills in use of a new system. This article looks at the maintenance phases in implementing a new system, using a changeover to problem-oriented recording as an example. Through this illustration, the essential elements for maintaining a high level of performance of a new skill are presented.


Kinetics of digestive enzyme stability in solid state I: application of Weibull distribution function to solid-state enzyme inactivation. The Weibull distribution function was applied to solid-state enzyme inactivation. On Weibull probability paper (within a narrow range), the plots of the accumulated inactivation ratio of each enzyme versus time regressed to a straight line. The parameters m and k, which correspond to the type and rate of the type and rate of the inactivation, were characteristic of each enzyme. The effect of temperature and parameter reproducibility are discussed.


Retinal disinsertion syndrome: report of a case. A case of retinal disinsertion syndrome in a 31-year-old mentally retarded patient was reported. Associated anomalies included subluxation of the lens, microphthalmos and bilateral keratoconus. Total retinal detachment and subluxation of the lens in a blind microphthalmic eye of a young infant should raise the possibility of retinal disinsertion syndrome.


Pancreatic elastase and serum alpha 1-antitrypsin levels in beagle dogs smoking high- and low-nicotine cigarettes: possible mechanism of pancreatic cancer in cigarette smokers. Beagle dogs exposed to cigarette smoke for 600 d experience a significant change in pancreatic elastase levels, as measured in tissue homogenates, compared with their sham-exposed controls. Greater elastase activity was found in the high-nicotine cigarette smokers than in the low-nicotine cigarette smokers. Levels of serum alpha 1-antitrypsin, an antiprotease capable of complexing the excess elastase, were also investigated. Animals smoking high-nicotine cigarettes had significantly lower serum alpha 1-antitrypsin activities than controls. Low-nicotine smokers showed alpha 1-antitrypsin activities that were not significantly different from those of controls. The importance of these observations is reinforced by a number of studies suggesting that proteases, their inhibitors, and an imbalance thereof may be related to the onset of neoplastic lesions. Studies have indicated that antiprotease levels follow the patterns of Mendelian inheritance. Severe deficiency states predispose human subjects to emphysema. A similar relationship may exist between antiprotease levels and susceptibility or resistance to neoplasms of the pancreas, a concept that deserves investigation in light of the findings reported here.


Acute purulent conjunctivitis in Nigerian children in Zaria. A prospective study of 61 acute purulent conjunctivitis seen in children up to 10 years of age at the Ahmadu Bello University Hospital, Zaria, Nigeria, between October 1975 and September 1976 showed H influenzae as the most common etiologic agent (26.2%), followed by S aureus (16.4%), N gonorrhoeae (14.8), and D pneumoniae (9.8%). N gonorrhoeae was the most common cause of conjunctivitis in the first month of life (43.8%) followed by S aureus (37.5%). Conjunctivitis from H influenzae occurred in older children, was more often bilateral, and had a male predilection. Gonococcal conjunctivitis occurred mostly in neonates and was largely unilateral. A good response to treatment with parenteral penicillin and topical Neosporin or chloramphenicol was noted in all the cases of gonococcal infection.


Congenital hereditary corneal dystrophy associated with esotropia. A family is described in which four of six siblings have congenital hereditary corneal dystrophy associated with esotropia. All cases had been erroneously diagnosed as and operated on for congenital glaucoma. The hereditary aspect of this association is discussed.


Effects of polarizing currents and repetitive stimulation on the uptake of amino acids by peripheral nerve. The effects of polarizing currents and repetitive stimulation on the uptake of representative amino acids have been studied in peripheral nerve. Glutamic acid, lysine, pheylalanine, and GABA were chosen for study as representatives of different carrier-mediated transport mechanisms. Depolarizing currents inhibited uptakes of glutamic acid, lysine, and GABA; maximum inhibitions were 56.1%, 20.0%, and 16.7%, respectively. Pheylalanine uptake was not inhibited by polarizing currents under any experimental conditions. Only glutamic acid uptake was inhibited by hyperpolarizing current, with a maximum inhibition of 43.9%. Stimulation had no significant effect on uptake of lysine, phenylalanine, or GABA but was found to potentiate uptake of glutamic acid by 12%.


Blood glucose and insulin levels, thyroid function, and serology in Ménière's disease, recurrent vestibulopathy, and psychogenic vertigo. Nineteen patients with unilateral Ménière's disease, 20 with psychogenic dizziness, and 20 with recurrent vestibulopathy (diagnostic criteria in text of paper) were found to have normal five hour glucose tolerance tests, serum thyroxine and effective thyroid indices, and serologic tests for syphilis. Hypothyroidism and hypoglycemia were absent in all groups. An unexplained finding of each diagnostic group was significant increase of fasting blood glucose and insulin levels, and elevated insulin:glucose ratios, compared to a control group. There appears to be no diagnostic indication for performing these chemical and serologic studies in patients with unilateral Ménière's disease, psychogenic vertigo, or recurrent vestibulopathy. Reasons are given to support the view that recurrent vestibulopathy may be a specific vestibular disturbance.


[Valve prosthesis in calcified aortic heart defects]. Experience in surgical correction of aortic valvular disease with extreme calcification of valves in 145 patients is generalized. The method for valve prosthetics preventing the development of specific complications is described. On the basis of analysis of the immediate and late postoperative results, it is recommended to use cold cardioplagia for protection of the myocardium. Hospital postoperative mortality was 13.9% in the last 4 years and 10.5% in the last 18 months. Preliminaly coronarography is suggested for the examination of patients with marked angina pectoris and with local changes on the ECG in the group of elderly patients for ruling out a concomitant organic lesion of the coronary artery.


[Change in the immunological indices in arteriosclerosis]. Fifty-nine patients with stage I (according to A. L. Myasnikov's classification) coronary atherosclerosis and 23 healthy persons of the same age were tested for the content of rosette-forming T- and B-lymphocytes, immunoglobulin classes, lymphocyte blast-cell transformation reaction to the phage and antigen from the normal and atherosclerotic aorta, and the changes in these indices under the effect of miscleron (in 23) and acetylsalicylic acid (in 10 persons). A tendency to an increase in the content of B-lymphocytes and G and A immunoglobulins and intensification of the lymphocyte blast-cell transformation reaction to an antigen from the sclerotic aorta, and a tendency to a decrease in the content of T-lymphocytes in patients with atherosclerosis were noted. The normalizing effect of miscleron therapy on these indices with parall improvement in the values of lipid metabolism were also established. Acetylsalicylic acid did not change the values of lipid metabolism and had not effect on the blood immunoglobulin content.