HLA antigens: rabbit antisera reacting with all A series or all B series specificities. Papain-solubilized HLA antigens giving only two bands of 34 000 mol.wt. and 11 000 mol.wt. by sodium dodecyl sulfate (SDS) gel electrophoresis and isolated from the cultured human lymphoblastoid cell line RPMI 4265, have been used to prepare antisera in rabbits. Antisera were raised against soluble products of the A (or 1 st) series of HLA, bearing the determinant HLA-A2 (A2 substance), or against a mixture of products of the B (or 2nd) series of HLA, bearing the determinants HLA-B7 and HLA-B12 (B7-12 substances). Rabbit antisera to A2 substance reacted primarily with A2 substance on Ouchterlony analysis, showing an apparent spur of cross-reactivity with B7-12 substances. Rabbit antisera to B7-12 substances reacted primarily with B7-12 substances, giving a spur of cross-reactivity with the A2 material. Neither antiserum precipitated beta-2microglobulin. Both types of sera reacted with membrane molecules of 43 000 mol.wt. and 11 000 mol.wt. by immune precipitation and gel electrophoresis in SDS of detergent-solubilized radiolabeled membranes from cultured cell lines, as predicted for sera directed towards HLA antigens. F(ab')2 fragments of the antibodies blocked the complement-mediated cytotoxicity of all HLA alloantisera tested for human peripheral blood lymphocytes. After absorption with B7-12 substances, F(ab')2 fragments of antisera to A2 substance only blocked the cytotoxicity of HLA alloantisera to A series specificities. After absorption with A2 substance, F(ab')2 fragments of rabbit antisera to B7-12 substances only blocked the cytotoxicity of HLA alloantisera to B series specificities. The results prove the existence of shared antigenic determinants between all members of the same series. These findings support the genetic evidence that A series HLA antigens are allelic products of a single locus, while B series HLA antigens are allelic products of a separate locus, by establishing some invariance of structure, presumably amino acid sequence, between members of the same series. The apparent cross-reactivity between A2 substance and B7-12 substances, and the ability of the unabsorbed F(ab')2 preparations to block the cytotoxicity of all HLA alloantisera, suggests that some determinants are common to both HLA loci. This may be considered to support the hypothesis that the two loci arose by gene duplication.


Structural and immunological similarities between HLA antigens from three loci. HLA antigens are coded for by three closely linked loci. The gene products of the first and second locus are known to be very similar in overall structure. Antigens coded for by the third locus were examined with regard to their chemical and immunological relationship to the first and second loci gene products. It is demonstrated that the third locus antigens are composed of two types of polypeptide chains, the smaller of which is identical to beta2-microglobulin. The alloantigenic polypeptide chain has an apparent molecular weight of 48 000 when solubilized by detergen treatment and about 35 000 when released from the cell membrane by proteolysis. These data together with size and shape analyses by gel chromatography and sedimentation velocity determinations show that HLA antigens from the three subloci are indistinguishable with the techniques employed. Suggestive evidence was obtained that the third locus antigens are more sensitive to proteolysis than the first and second locus antigens. The similarity between the three gene products was also evident on cyanogen bromide fragmentation of the polypeptide chains. Furthermore, rabbit antibodies against antigens coded for by the first locus cross-reacted extensively with antigens from the second and third locus. This study therefore lends strong credence to the view that the three HLA antigen subloci have arisen by gene duplications of a common ancestral gene.


Dantrolene and the neuromuscular junction: evidence for intracellular calcium stores. Dantrolene sodium (DaNa) markedly depressed the frequency of spontaneous miniature potentials at the frog neuromuscular junction. Its action is still observed at low [Ca2+]o and is little affected by temperature; its effect is not readily reversible. Theophylline antagonises the action of DaNa. DaNa prevents the stimulatory effect of the ionophore A23187 at 25 degrees C, but is ineffective in antagonising the action of X537A. DaNa and theophylline do not affect the functioning and integrity of mitochondria in vitro and it is suggested that DaNa acts at intracellular Ca2+ stores (other than the mitochondria) in the presynaptic terminals.


Release of renal prostaglandins during endotoxin-induced hypotension. The appearance of prostaglandins in dog's blood during endotoxin-induced hypotension was studied by use of the dialysis modification of the blood bathed organ technique. An increase in prostaglandins, mainly E2 and F2alpha was found in renal venous blood, whereas no such increase was seen in blood from the abdominal aorta, the inferior vena cava or the femoral vein. Three possible trigger mechanisms for this increase i.e. hypotension, reduced flow and reflexogenic sympathetic stimulation, have been investigated. It is suggested that, in addition to these three factors, circulating hormones such as noradrenaline, angiotensin or bradykinin, play a role in this release mechanism. Administration of indomethacin produced a restoration of the systemic blood pressure to its pre-endotoxin value; concomitantly a disappearance of the prostaglandins from the circulation was observed. It is concluded that prostaglandins contribute to the hypotension induced by endotoxin. Whether they are beneficial or detrimental remains to be resolved.


[Ribonuclease activity following partial hepatectomy in the liver of healthy and alloxan diabetic rats and histological findings in the islets of Langerhans (author's transl)]. What changes in the level of ribonuclease activity (RNase activity) can be observed in the regenerating liver of healthy and alloxan diabetic rats following partial hepatectomy? What coincidental alterations of the ratio A-cells to B-cells do occur in the islets of Langerhans? 120 male Wistar rats of averagely 180 g body weight were fed a standard diet ("Rehbrücke") with drinking water ad libitum. After deprivation of food for 15 hours the animals were sacrificed by heart puncture under ether anesthesia. The caudal part of the body was rinsed free from blood by ice-cooled physiological saline via the aorta. After removal of the livers each 2 g of liver tissue were homogenized in 0.25 M saccharose or 70% ethyl alcohol. For the study of impairment of ribonuclease activity in the liver following different lesions the animals were divided into 3 test groups of 35 animals each: group 1: alloxan diabetes; group 2: liver regeneration after partial hepatectomy; group 3: alloxan diabetes in combination with liver regeneration after partial hepatectomy. From each group always 4 to 6 animals were sacrificed at the following time intervals: 2nd day and 4th day as well as after 3, 4 and 6 weeks. In 8 animals the enzymatic activity was not influenced. RNA content and dry weight of the livers of all test animals were determined. In liver homogenates with saccharose the ribonuclease activity was measured according to the method of FIERS (1961) using highly purified yeast RNA for substrate. This technique is especially suitable for raw tissue homogenates. After 30 min incubation at 37 degrees C and precipitation of the high-molecular polynucleotides as well as of protein by means of methylglycol and barium perchlorate the not precipitable low-molecular oligonucleotides which were produced by ribonuclease action, were spectrophotometrically measured in the supernatant at 260 nm. The deltaE-value was calculated for the employed quantity of RNA related to mg of protein brought into action. Demonstration and determination of the ribonuclease inhibitor was abandoned. Two different RNA substrate concentrations were used; in the higher concentration the activity was measured at pH 5.5 to 6 and 7.5, in the lower concentrations at pH 7. Protein measurement was performed after the method of LOWREY (LOWREY et al. 1951) as modified by GLASER and KLEINE (1962); blood sugar was determined in tail vein blood by means of o-toluidine technique. In the liver homogenate with ethyl alcohol the RNA extraction was modified according to the data given by OGUR and ROSEN (1950); RNA determination was done colorimetrically using orcin hydrochloric acid. For dry weight determination the same homogenate was used. Alloxan diabetes was induced by injecting the rats 175 mg/kg b.w. of alloxan after fasting for 24 hours. The 5% alloxan solution in citrate buffer (pH = 4) was prepared immediately before injection. For further experimentation only those animals with blood sugar values of greater than 300 mg/100 ml were employed...


[Electron microscopic investigation following canine pulmonary transplantation with regard to functional effects (author's transl)]. The function of alveolar macrophages in correlation with immunological stimulation, consequently also in pulmonary transplantation, are a subject of discussion. In this study the alveolar manifestations of rejection following pulmonary transplantation, are a subject of discussion. In this study the alveolar manifestations of rejection following pulmonary homotransplantation are investigated by electron microscopy. In 20 healthy dogs (body weight between 7 and 19 kg) a left side pulmonary homotransplantation was performed using a standardized method. 2 randomized groups of 10 animals each were formed, one undergoing immunosuppressive therapy, the other none. Needle biopsies (Menghini needles) were taken in both groups at the onset of rejection daily until death of the animals (on the 5th to 12th postoperative day). At death of the animals always total rejection of the homograft was observed. The other animals were subjected to a scheduled immunosuppressive therapy (KREUZER et al. 1973, RADASZKIEWICZ et al. 1974). Needle biopsies were taken only at the stage of a clinically or roentgenologically ascertained crisis of rejection. Before implantation of the homograft and 20 mins after occlusion of anastomoses and onset of circulation in the transplant biopsies were taken, too, to exclude early changes or alterations in the donor. The lung specimens 2 mm in size) obtained by biopsy were fixed in phosphate buffered OSO4 and buffered phosphate solution by usual alcohol series. After embedding in epon the specimens were studied by electron microscopy. In the group not undergoing immunosuppressive therapy beginning with the 3rd post-operative day perivascular lymphocytic and plasmacellular infiltrates were seen which increased during the course of rejection. In both groups a swelling and vacuolization of endoplasmatic reticulum, cytoplasmic microvilli with bridge formation accompanied by an increase of alveolar macrophages and by augmentation of lamellar bodies in granular pneumocytes were noted following operation. A correlation between the increase of lamellar bodies and increase of surfactant has been shown previously.


Growth inhibition and morphological changes caused by indomethacin in fibroblasts in vitro. In a definite range of concentrations indomethacin inhibits the multiplication of fibroblasts in vitro without influencing their viability. The effect of lethal doses of indomethacin on the fine structure of normal and transformed fibroblasts was investigated by light and electron microscopy. The cells grown in flat monolayers became roundish and, varying in sequence, small zeiotic blebs were bulging at the external cell membrane. Beside coarse vesicular transformation of the endoplasmatic reticulum and extension of the perinuclear spaces ballooning of the mitochondria occurred. After breaking up of the vesicles of the endoplasmatic reticulum loosening of the cytoplasmic ground substance associated with enlargement of the hyaline plasma margins was observed in further progress of the experiment. The internal structures of the mitochondria were completely disorganized. In the ultrastructure of the fibroblasts altered by lethal doses of indomethacin no specific sites of drug action could be identified.


Protein synthesis and secretion in the cobalt chloride damaged exocrine pancreas of guinea pigs. An electron microscopic-autoradiographical and histochemical study. The cobalt chloride induced disturbance of secretion in the pancreatic acinar cells of guinea pigs consists, on the one hand, in a continued but reduced synthesis of exportable enzyme proteins, on the other hand in an alteration of both function and structure of the Golgi apparatus and the mitochondriae, followed by a loss of zymogen granules and the occurence of numerous intracisternal granules. The latter are transported through the cisternae of the endoplasmic reticulum to the apical cell pole and discharged into the lumen of the acinus without any interposition of the Golgi apparatus.


The use of fluorescein isothiocyanate labelled lectins for immunohistochemical demonstration of saccharides. Cryostat sections of various tissues from man, rat, mice, hamster, rabbit, guinea pig, sparrow and carp were examined by fluorescence microscopy after incubation with fluorescein isothiocyanate labelled Concanavalin A and Lens culinaris lectin. In the same tissue of various species no basic differences in the pattern of fluorescence were observed. In general, connective tissue structures were stained by both lectins in the same degree, whereas by fluorescein isothiocyanate labelled Concanavalin A additionally a varying staining of the cell surface and of cytoplasmic structures occurred. The results show the general presence of Concanavalin A and Lens culinaris lectin receptors in normal animal tissues.


Electron microscopy of Kupffer cells in the orthotopic porcine liver homograft during the late stage after transplantation (phagocytosis of host cells by Kupffer cells). Electron microscopical observations of macrophages (Kupffer cells) in the orthotopic porcine liver homograft in later stages after transplantation are described and demonstrated. The findings show close topographical relations between the Kupffer cells and hepatocyte debris, erythrocytes, thrombocytes, lymphocytes and granulocytes. Besides an uptake of hepatocyte debris, a phagocytosis and degradation of apparently unaltered erythrocytes, thrombocytes and probably also  lymphocytes by the macrophages were observed. The possible significance of these findings is discussed with regard to the immunological interactions between macrophages and lymphocytes, to the course of the rejection in the liver homograft, to the effect on the blood cells and platelets of the host organism, and thus to the long-term prognosis of liver transplantation.


[Experimental xenogenic immune pancreatitis. --Immunohistological, enzyme histochemical and ultrastructural studies (author's transl)]. Repeated intraperitoneal injections of anti-mouse pancreas rabbit serum or of anti-mouse pancreas guinea pig serum produce a chronical sclerotizing pancreatitis. This study has the aim to contribute to the further elucidation of the changes which occur in the acinar cells, as well as to the etiology and pathogenesis of immune pancreatitis, by means of immunohistological, enzyme histochemical and electron microscopic studies. Anti-mouse  pancreas rabbit serum was obtained by sensitization of rabbits with an admixture of AB-mouse pancreas extract (100,000 g - supernatant) and complete Freund's adjuvant [details see NIZZE, Exp. Path. (1975a)]. The presence of precipitating mouse pancreas antibodies in the rabbit serum was ascertained by the agargel diffusion test according to Duchterlony (1958). The experiments were performed with 54 adult male white mice (AB colony strain) of 22 to 30 g.b.s. (averagely 26 g). The animals were divided into 4 groups which were treated as follows: 1. 24 mice with anti-mouse pancreas rabbit serum, 2. 12 mice with rabbit normal serum, 3. 12 mice with physiological saline, 4. 6 mice remained untreated (controls) Always 4 animals of the group 1 as well as each 2 of the groups 2 and 3 were administered in total 1, 3, 5, 9, 17 or 33 intraperitoneal injections of 0.3 ml of the correspondent serum or with physiological saline within 3 hours, 1, 2, 4, 8 or 16 days. The last injection was regularly applied 3 hours before sacrification by decapitation. The time of sacrification was always at 11.00 o'clock a.m. For immunohistological and enzyme histochemical investigations 10 mum thick cryostat sections were prepared consisting of pancreatic specimens piled up to a bloc. In each case the tissue samples  were taken from the experimental animals and from one control animal sacrificed at the same day. The sections were incubated in FITC-labelled anti-rabbit globulin goat serum at room temperature for 30 min in a moist chamber. For control of specificity were employed: a) initial incubation of equal sections with unlabelled anti-rabbit globulin goat serum for 30 min (""blocking test''), b) pancreatic tissue specimens of each one untreated control animal present in the cryostat sections and thus incubated like the pancreatic tissue of the experimental animals, c) native nonincubated cryostat sections from the same bloc to exclude nonspecific autofluorescence. Evaluation of the sections was done in a Zeiss-Lg-microscope with HBO-50 high pressure mercury lamp. Exciter filters were UG 1/3.5 and 1/1.5, the eyepiece was screened with a GG 9/1 filter photographs were taken on ORWO X-ray film RS 2 (VEB Filmfabrik Wolfen). The enzyme histochemical studies were performed on cryostat sections of the same tissue bloc using the following methods: lead nitrate- or calcium-Co-method after GOMORI (1952) for demonstration of acid and alkaline phosphatase, naphthylacetate method (NACHLAS and SELIGMAN 1945) for nonspecific esterase, MTT-co-method (PEARSE et al...


Electron microscopy of lymphocytes and hepatocytes in orthotopic liver homografts during the late rejection (target cell destruction by lymphocytes in vivo?). The results of electron microscopical observations made on orthotopic procine liver homografts during the process of delayed rejection show close contacts between the cell membranes of lymphocytes and hepatocytes, the lymphocytes reaching the hepatocytes by incomplete and complete migration through the partially enlarged intracellular and intercellular gaps of the endothelial sinusoid lining. In addition, the lymphocytes are capable of coming into contact with the surfaces of hepatocytes through both intercellular and intracellular endothelial gaps by means of microvilli form and uropodlike cytoplasmic projections. The hepatocytes exhibit partly direct, partly indirect signs of a localized change of the cell membrane which is associated with abnormal permeability, partial peripheral lysis and colliquation necroses. Such alterations were frequently found to be located in the immediate neighborhood of both lymphocytes and lymphoid cells. The interpretation of these electron microscopical findings as being an expression of in vivo target cell destruction by lymphocytes is discussed.


Chloral hydrate induced haploidization in Aspergillus nidulans. This is the first report of induction of haploidization in Aspergillus nidulans by chloral hydrate, which is an efficient polyploidizing agent for higher plants and a psychotropic drug for man. A new procedure has been described to isolate haploids from diploids with a very high frequency, as compared to p-fluorophenylalanine, which is generally used for this purpose.


Immunosuppressive effect of a mouse placenta fraction on H-2 incompatible split heart allografts. A soluble placenta fraction from mice (A.CA) mated with H-2 histoincompatible males (A/Sn) significantly prolonged the survival of heterotopic A/Sn heart transplants in A.CA recipients. No prolongation of A/Sn heart graft survival was obtained with the corresponding A.CA placenta fraction after A.CA X A.CA mating.


Experimental autologous immune deposit nephritis in rats associated with mercuric chloride administration. Serial administration of mercuric chloride to rats was followed by development of antibodies to tubular basement membrane and renal tubular epithelial antigen (RTE) and glomerulonephritis characterized by granular deposits of hosts IgG, C3 and RTE along the glomerular capillary walls. The glomerular fixed antibody was directed against RTE. These studies suggest that tubular injury by mercury may lead to release of RTE and autosensitization and subsequent antibody production to this antigen result in formation of and glomerular deposition of circulating immunopathogenic complexes (RTE-anti-RTE) and glomerular morphologic alterations.


Pineal gland biorhythms: N-acetyltransferase in chickens and rats. Circadian rhythms have been identified in chemical factors involved in the metabolism of serotonin in the pineal gland. The rhythms are reviewed with respect to their characteristics and regulation by environmental lighting in vertebrates. Special emphasis has been given to the rhythm in pineal N-acetyltransferase activity in rats and chickens. This rhythm is not only apparent in a light-dark cycle in some birds and mammals, it also persists in constant dark. In constant light the rhythm is lost. The action of light is via the eyes in rats but extraretinal photoreceptors are involved in chickens. The details of the N-acetyltransferase activity rhythm include the presence of a refractory period for the rise in dark-time enzyme activity and the rapid turnoff of high dark-time activity by light. The N-acetyltransferase clock can be reset with a period of constant light in the chicken. Neural regulation of the rhythm in rats involves the suprachiasmatic nucleus and the superior cervical ganglion; in chickens, the ganglion is not necessary for the rhythm in light-dark cycles and the suprachiasmatic nucleus has not been studied.


Membranes and molecules in circadian systems. A membrane clock model involving ions and ion transport is compatible with the limit cycle concept, biochemical studies, and the idea of phase shifting by ion gating. It provides an explanation for temperature compensation, and, by assuming that oscillating ion concentrations regulate the many different biochemical and physiological rhythms, it explains how diverse systems can be controlled by a single oscillatory mechanism. How ions control ion transport and which membranes are involved in the clock are two questions we have begun to explore.


Human circadian rhythms: a multioscillatory system. After a brief review on temporal order within the human organism and implications for chronopharmacology, the paper discusses evidence indicating that the human circadian system consists of a multiplicity of oscillators. Subjects who live in isolation without time cues show free-running circadian rhythms deviating from 24 hours. Often, the activity rhythm (wakefulness and sleep) and other rhythmic variables (e.g., temperature) have the same circadian period of about 25 hours (referred to as the state of internal synchronization) but on occasions the activity period may become substantially longer (e.g., 33 hours) while the other rhythms continue with a period of about 25 hours. Such a state is termed internal desynchronization. There are also cases where the activity rhythm can reach extreme values of about 50 hours; to such a circa-bi-dian rhythm other variables are again synchronized, but in a 2:1 ratio. Internal desynchronization can occur both by shortening and by lengthening the activity rhythm. In these two cases the periods of other rhythms also change slightly in a direction opposite to that of the activity rhythm, indicating a loss of coupling between two classes of basic oscillators that both influence the two groups of overt rhythms but by different extents.


Prostaglandins and local circulatory control. The present paper reviews several lines of investigation have provided highly suggestive evidence for an important regulatory role for prostaglandins in the microcirculation. Aside from their profound vasodepressor effects in a number of animal species, including man, and their vasodilator activity in many local circulatory beds, endogenously administered prostaglandins of the E and A type also appear to reduce vascular responsiveness to a variety of vasoconstrictor agents. Furthermore, inhibition of endogenous synthesis and release of prostaglandins leads to a potentiation of vasoconstrictor responses. Thus it appears that the release of prostaglandins, an event that accompanies or is a consequence of vasoconstriction, can moderate the constrictor response and in this manner can contribute to the control of vascular reactivity. Administration of inhibitors of prostaglandin synthesis to tissues that under basal experimental conditions release prostaglandins, causes not only a decrease in prostaglandin output but also an increase in resting perfusion pressure. These results support the concept that prostaglandins that are released are essential in the control of vascular tone and resistance. Other evidence also suggests that prostaglandins participate in a variety of vascular responses, including the mediation of bradykinin vasodilation, functional hyperemia, and reactive hyperemia.


The synthesis and function of prostaglandins in the heart. The isolated perfused rabbit heart has the capacity to rapidly synthesize, release, and inactivate prostaglandins (PGs) which can readily modulate coronary resistance and cardiac performance. Prostaglandin synthetase converts arachidonic acid to prostaglandin-like substance in one passage across the heart. There is little evidence of functional prostaglandin dehydrogenase activity. Arachidonic acid and bradykinin produce a concentration-dependent decrease in coronary resistance directly associated with PG-like substance biosynthesis and release. An inhibitor of bradykinin destruction, the nonapeptide SQ-20881, markedly enhanced both the coronary vasodilation and PG-like substance release produced by cardiac injection of bradykinin. Indomethacin inhibited both the myocardial prostaglandin biosynthesis and the decrease in coronary resistance induced by bradykinin and arachidonic acid. In addition to direct effects on coronary vascular smooth muscle, prostaglandins produced in the heart apparently exert a modulating influence on efferent autonomic and on afferent cardiovascular and pain reflexes.


Prostaglandins as determinants of vascular reactivity. Prostaglandins are primarily local or tissue hormones that have their effects at, or near to, the site of synthesis. Some blood vessels synthesize prostaglandins intramurally, where their local release influences vascular tone and reactivity. Endogenous prostaglandins (primarily prostaglandin E2 (PGE2) participate in the regulation of vascular reactivity by opposing the vasoconstrictor and antinatriuretic actions of circulating pressor hormones; and by braking the release of norepinephrine from vasoconstrictor nerves. The proposal that one or more prostaglandins affect vascular reactivity is supported by the following observations: enhanced vascular reactivity to pressor stimuli occurs in organs with low basal rates of prostaglandin synthesis and after inhibition of prostaglandin synthetase in organs with high biosynthetic capacity; and exogenous PGE2 reversibly inhibits the vasoconstrictor activity of pressor stimuli.


Pituitary gonadotropin response in women with idiopathic hyperprolactinemia. Inappropriate lactation and idiopathic hyperprolactinemia are frequently associated with amenorrhea. In these individuals, peripheral levels of follicle-stimulating hormone (hFSH) are usually normal, and luteinizing hormone (hLH) levels are often found in the low-normal range. The present study was undertaken to evaluate the functional capacity of the pituitary by the response of hFSH and hLH to synthetic gonadotropin-releasing hormone (Gn-RH). Six women with amenorrhea, inappropriate breast secretion, and idiopathic hyperprolactinemia (prolactin levels ranged from 45 to 355 ng/ml) were given 100 mug of Gn-RH intramuscularly. Serum hFSH and hLH levels were assessed in samples obtained at 15-minute intervals over the next 2-hour period. Initial hFSH levels were normal in all women, with a mean of 242 +/- 72 ng/ml. The absolute increase after Gn-RH administration averaged 486 +/- 193 ng/ml. Serum hLH was below normal in three of the six women, and normal in the remaining three women initially. The absolute increase averaged 1308 +/- 315 ng/ml. The greatest percentage increase in hLH was found in the women with the subnormal basal titers. In these women, hLH rose from a mean of 22 ng/ml to a mean of 1092 ng/ml. These data demonstate an exaggerated increase in hFSH and hLH levels after exogenous Gn-RH administration. This suggests that the amenorrhea associated with elevated serum prolactin levels is principally of hypothalamic origin.


Effects of a luteinizing hormone-releasing hormone analog on mating and fertility in rats. A rhythmic antifertility effect of a luteinizing hormone-releasing hormone (LH-RH) analog, [D-Ala6-des-Gly-NH210]-LH-RH-ethylamide (AY-25,205), administered intramuscularly every 3rd day staring in the afternoon of diestrus, was demonstrated in 4-day cyclic rats. The antifertility effect was achieved for a period of approximately four cycles when the females were allowed constant cohabitation with fertile males except for 24 hours following treatment. Unrestricted cohabitation resulted in some matings and pregnancies in the group treated every 3rd day and also in some of the groups treated every 4th day with restricted cohabitation. The antifertility effect of AY-25,205, with unrestricted cohabitation, disappeared when the second treatment was given 4 days after the first. It is presumed that the antifertility effect of AY-25,205 was achieved through its capacity to induce ovulation at a physiologically "wrong time" (i.e., 1 day before the expected day of proestrus) and through its effect on mating behavior. The present experimental models suggest that AY-25,205 or similar analogs could be potentially useful for a more reliable rhythm method of birth control in humans, by timing ovulation and narrowing the fertile period of the cycle.


Haemoglobin as a marker for bone marrow transplants in laboratory rats. We made use of the genetic polymorphism of haemoglobin in the laboratory rat and developed a congenic strain LEW.Hb A differing from the original LEW strain in the type of haemoglobin. The exclusion of the histoincompatibility reactions allows a long-term follow-up of the bone marrow transplants on the basis of changes in the haemoglobin pattern.


Quantitative aspects of abolition of transplantation tolerance by the adoptive transfer of lymphoid cells. It was found that with neonatally induced transplantation tolerance about 50% of the rats are insensitive, or little sensitive, to the adoptive transfer of immunity. The proportion of rejected and unrejected skin grafts in individual experiments did not show a direct dependence on the dose of transferred cells, and immune lymphocytes were as effective as non-immune cells. No dependence on the time post-transplantation was found. Only the results of the adoptive transfers performed after more than 120 days after transplantation indicated an increased resistance of the tolerant state. Also repeated transfers in which increasing doses of cells were used showed a relatively stable state of tolerance in some animals. It is discussed that the difference in the sensitivity to the adoptive transfer might reflect at least quantitative individual differences in the mechanism of the state of unresponsiveness in neonatally induced tolerance.


Influence of thymosterin on immunological reactivity in New Zealand mice. The influence of thymosterin on immunological reactivity of NZB mice was investigated. Thymosterin was administered three times per week for a period of six weeks to male and female NZB mice. Only lymphocytes from untreated female mice gave the positive results in the macrophage migration inhibition test using native DNA as the stimulating antigen. This effect was suppressed in thymosterin treated females. The administration of thymosterin therefore appears to correct the in vitro delayed hypersensitivity to DNA antigen.


[Effect of somatostatin on pancreatic endocrine--experiment by somatostatin infusion into the pancreaticoduodenal artery in dogs--(author's transl)]. The direct inhibitory action of somatostatin (cyclized SRIF) on the pancreatic endocrine function was investigated by a technique using pancreaticoduodenal arteriovenous system in vivo. Somatostatin was infused for 20 minutes at a speed of 2.5 mug/minute into the superior pancreaticoduodenal vein and femoral artery were measured before and every 5 minutes throughout the experiment for 30 minutes. The results were compared with those obtained from the control experiment which was carried out in the same time schedule under infusion of physiologic saline solution instead of somatostatin. Next, the effect of somatostatin on the glucose-induced insulin release from the pancreas was also evaluated. The following findings were obtained. (1) Somatostatin infused at a speed of 2.5 mug/minute into the superior pancreaticoduodenal artery caused a statistically significant inhibition of plasma levels of IRI and IRG and  also pancreatic output of these hormones. With the sessation of somatostatin infusion an abrupt rise of the hormones were seen. This "rebound" phenomenon was more pronounced in insulin secretion than in glucagon. No significant changes in the plasma glucose levels in either the pancreaticoduodenal vein or the femoral artery throughout the experiment were found. (2)During infusion of somatostatin at a speed of 1.25 mug/minute, insulin response to glucose injected into the pancreaticoduodenal artery in a small dose, as well as having no effect on the plasma glucose level in systemic circulation, was also significantly inhibited. From these findings obtained by direct experiment using the pancreaticoduodenal arterio-venous system, it was confirmed that somatostatin exhibits a direct inhibitory action on pancreas endocrine in a very low concentration in vivo, and it was suggested that this action might be partly due to a reduction in pancreatic circulation.


[The trophedema (Nonne-Milroy-Meige). Carcinogenesis as a rare complication]. The trophedema Nonne-Milroy-Meige has an exceptional position within the group of the primary lymphatic edemas (l.e.) because of its hereditary. Its frequency less than 1% of primary l.e. The trophedema is caused by a genetic determined defect of the morphogenese of parts of lymphatic system, which is mainly autosomal dominantly transmitted. It is morphologically and lymphografically characterized by a lack and reduction respectively of the number of lymphatic vessels. The trophedema results an emotional (cosmetic) and physical stress. Complicationes will rarely arise. In this paper it is described the case of the development of a cancer upon a trophedema, which seems to be the first case ever published. It will be shown, that an test-section have to be carried out in all cases of damages at a l.e. als soon as possible. The best conservative method at present used is the treatment with cortisone and hyaluronidase including bandage. However a real cure of the primary l.e. including the trophedema can not be attained by therapeutic methods presently used, because the defect of the lymphatic-vessels-system is hereditary. On the other hand therapeutic nihilism cannot be recommended.


Basement-membrane thickening and diabetic microangiopathy. This report considers the pathophysiologic significance of capillary basement-membrane thickening in diabetic nephropathy and retinopathy and the relationship of capillary basement-membrane thickening to increased susceptibility to infections and to increased vascular permeability in diabetes. The evidence available (1) indicates that basement-membrane thickening affects most if not all capillaries of the diabetic and may contribute to increased susceptibility to infection and (2) suggests that increased capillary permeability in diabetes need not be attributed to basement-membrane changes per se, but rather may be due to changes in the cellular elements of the capillary wall.


Passive immunotherapy in HBs Ag fulminant hepatitis. Results on antigenaemia and survival. 19 patients with HBs Ag-positive fulminant hepatitis underwent exchange transfusions and anti-HBs-rich plasma infusions. Circulating HBs Ag disappeared in 17 cases. Circulating anti-HBs detected by RIA was higher in cases with liver regeneration. There was no correlation between circulating anti-HBs level and the amount of anti-HBs-rich plasma administered. Recovery was observed in 36% of the cases and liver regeneration in 47%. As the beneficial effect of such a treatment is not yet evident, a controlled trial should require pairing of patients with respect to the number of exchange transfusions and amount of real or placebo anti-HBs.


Calcium induced glucagon release in monolayer culture of the endocrine pancreas. Studies with ionophore A23187. The possible role of Ca2+ in glucagon release has been investigated by the use of ionophore A23187. This ionophore permits Ca2+ entry down a suitable concentration gradient by complexing and releasing Ca2+, thereby acting as a carrier in plasma membranes. Cultured cells obtained by enzymatic digestion of pancreases from newborn rats were studied on the third day of culture. As expected the effects of the ionophore were dependent upon the presence of Ca2+ in the medium. However, either stimulation or inhibition of glucagon release resulted when different concentrations of ionophore and Ca2+ were used. With 1.0 mM Ca2+ in the medium, glucagon release was stimulated in the presence of 0.01 and 0.1 mug/ml ionophore, but inhibited in the presence of 3.0 and 10.0 mug/ml. With 0.1 mug/ml ionophore, glucagon release was stimulated by 0.3 and 1.0 mC Ca2+ but not by 2.5 mM Ca2+. With 10 mug/ml ionophore glucagon release was stimulated by 0.03, 0.1 and 0.3 mM Ca2+, whereas at 1.0 mM, glucagon release was depressed. These findings suggest that by increasing Ca2+, glucagon is released from the A-cells, whereas too large an increase in Ca2+ is inhibitory. The effect to stimulate release was not completely specific for Ca2+ in that while the ionophore did not stimulate release in the presence of either Mg2+ or Sr2+ in the absence of Ca2+, it did stimulate release when Ba2+ was tested. Furthermore Ba2+ at 0.3 mM was stimulatory even in the absence of ionophore. Glucagon release in the absence of ionophore was also enhanced by addition of 30 mM Ca2+ or by omission of Ca2+ from the medium. It is concluded that Ca2+, which plays an essential role in the stimulus-secretion coupling in several different cell types, may be involved in the stimulation of glucagon release from the A-cells of the pancreas.


Enhanced glucagon secretion by pancreatic islets from prednisolone-treated mice. This work was undertaken to study the effect of prednisolone on glucagon release in mouse pancreatic islets isolated by the collagenase technique. Pretreatment of the donors with prednisolone (0.2--0.3 mg daily) induced an increase in glucagon release both in the absence (1005+/-75, SEM, vs. 796+/-46 pg/10 islets/60 min, p=0.019) and in the presence of 7.5 mM arginine (1500+/-119 vs. 1236+/-61 pg/10 islets/60 min, p=0.05). The glucagon content of the islets was not modified by the treatment (28.6+/-1.1 vs. 28.0+/-1.1 ng/50 islets). The addition of prednisolone (5 - 10(-5) M) into the medium, failed to affect significantly glucagon secretion. In agreement with previous human studies, our data indicate that chronic glucocorticoid administration augments the secretory activity of the A-cell. This does not seem to be a result of increased glucagon synthesis nor a direct effect of glucocorticoids on the glucagon-releasing mechanism. Rather, environmental changes induced by these hormones could be responsible for A-cell hyperfunction.


Effect of sodium amylosulfate (Depepsen) on the healing of duodenal ulcer. Thirty-five patients with active duodenal ulceration were included in a double blind randomized trial, the antiulcer agent sodium amylosulfate (Depepsen), being compared with placebo. Diagnosis and healing were determined by duodenoscopy. Results showed, in the whole series, 13 of 18 patients treated with Depepsen healed, whereas 10 of 17 healed on placebo (P = 0.04). In the group of 23 outpatients, 6 of 11 healed on Depepsen, and 5 of 12 healed on placebo (P = 0.55). It was concluded that Depepsen did not accelerate the healing of duodenal ulcer.


A double blind crossover study of metoclopramide versus placebo for facilitating passage of multipurpose biopsy tube. Intravenous metoclopramide (M) was compared to placebo (P) by a double blind crossover design to determine whether M was superior to P in difficult cases of intubation of the small intestine, using a multipurpose biopsy tube and capsule. Metoclopramide decreased intubation time in 20 volunteers successfully intubated with M and P (P less than 0.05). Of 9 subjects, 8 were intubated to ligament of Treitz with M but not with P (P less than 0.01). Of 29 volunteers, 22 were successfully intubated by 15 min when M was given, but only 9 of the 29 could be intubated in 15 min with P (P less then 0.001). Of 29 volunteers receiving M, 9 experienced side effects but none were serious. This study demonstrated that M is superior to P in decreasing intubation time of a small intestinal biopsy capsule and is particularly useful in patients who may not otherwise be sucessfully intubated.


Thialysine-resistant mutant of Salmonella typhimurium with a lesion in the thrA gene. A mutant of Salmonella typhimurium was selected for its spontaneous resistance to the lysine analog, thialysine (S-2-aminoethyl cysteine). This strain, JB585, exhibits a number of pleiotropic properties including a partial growth requirement for threonine, resistance to thiaisoleucine and azaleucine, excretion of lysine and valine, and inhibition of growth by methionine. Genetic studies show that these properties are caused by a single mutation in the thrA gene which encodes the threonine-controlled aspartokinase-homoserine dehydrogenase activities. Enzyme assays demonstrated that the aspartokinase activity is unstable and the threonine-controlled homoserine dehydrogenase activity absent in extracts prepared from the mutant. These results explain the growth inhibition by methionine because the remaining homoserine dehydrogenase isoenzyme would be repressed by methionine, causing a limitation for threonine. The partial growth requirement for threonine during growth in glucose minimal medium may also, by producing an isoleucine limitation, cause derepression of the isoleucine-valine enzymes and provide an explanation for both the valine excretion, and azaleucine and thiaisoleucine resistance. The overproduction of lysine may confer the thialysine resistance.


F' plasmids from HfrH and HfrC in recA- Escherichia coli. We have isolated and characterized a number of clones resulting from matings of HfrH and HfrC cultures of Escherichia coli with auxotrophic recA- E. coli. As in Low's (1968) experiments, the recA- marker prevented integration of F' episomes into the vegetative chromosomes of the host. Both F'H F'C plasmids contained a great variety of non-selected nutritional markers. However, more F'H plasmids seemed to have expressed F+ characteristics than did F'C plasmids. These characteristics include (i) the presence of F-pili as determined by susceptibility to male-specific phages; (ii) fertility as determined by the merozygote's ability to transfer nutritional markers to an auxotrophic F- strain of E. coli; and (iii) a high degree of inheritability as estimated by the proportion of F' bacteria to F- bacteria in clones grown in a non-selective medium like broth. This proportion is seen to be affected by both factors that determine the probabilities that daughters of F' bacteria inherit the episome and from physiological factors that determine the rates of growth of F' and F- bacteria.


Mutator activity of petite strains of Saccharomyces cerevisiae. Petite strains in Saccharomyces exhibit enhanced spontaneous mutation rates of nuclear genes regardless of whether they are cytoplasmically or nuclearly inherited, or whether or not the cytoplasmic petite strains have mitochondrial DNA. In petite strains, the mutation rate for the nonsense allele lys1-1 is enhanced by a factor of 3-6 and for the missense allele his1-7 by a factor of 2 as compared with their grande counterparts. The reversion of a third allele, the putative frameshift mutation, hom3-10, is not enhanced in a petite background. The results indicate that the spontaneous mutation rate of an organism can be altered by indirect intracellular influences.


Mutators in Saccharomyces cerevisiae: MUT1-1, MUT1-2 and MUT2-1. The purpose of this study was to characterize two mutator stocks of yeast which were induced and selected on the basis of high spontaneous reversion rates of the suppressible "ochre" nonsense allele lys1-1. In the mutator stock VA-3, a single mutation, designated mut1-1, is responsible for the increase in the reversion rate of the ochre alleles lys1-1 and arg4-17. In stock VA-105, there are two separate mutator mutations. Tetrad analysis data showed these two loci are loosely linked. Based on complementation data, one of these mutations is at the same locus as mut1-1 and designated mut1-2. The second mutator of stock VA-105 was designated mut2-1. All three mutators are recessive. Both mut1-1 and mut1-2 give a high mutation rate for ochre nonsense suppressor (SUP) loci, but not for the ochre nonsense alleles. On the contrary, the mutation rates of the ochre alleles are greatly reduced. With the mutant mut2-1 there were mutations at both the lys1-1 site and its suppressors; mut2-1 is as effective as mut1-2 but not as effective as mut1-1 in inducing reversions of a missense mutant, his1-7. Neither mut1-1, mut1-2 nor mut2-1 were effective in inducing reversions of a putative frameshift mutation, hom3-10, or in inducing forward mutations to canavanine resistance.


Mapping of trichodermin resistance in Saccharomyces cerevisiae: a genetic locus for a component of the 60S ribsomal subunit. Resistance to the protein synthesis inhibitor trichodermin in Saccharomyces cerevisiae has been studied. A single recessive nuclear gene was responsible for resistance. The resistance locus, tcm1 was found to be closely linked (1 centi-morgan) to the locus pet 17 on the right arm of chromosome XV. The mutation to trichodermin resistance conferred resistance to other 12,13-epoxytrichothecenes and to the structurally unrelated antibiotic anisomycin.


Temperature-sensitive yeast mutants defective in meiotic recombination and replication. A system is described for isolating temperature-sensitive mutants of Saccharomyces cerevisiae with defects in early meiotic events. We used an otherwise haploid strain disomic (n+1) for chromosome III, and heteroallelic at the leucine-2 locus. Meiotic development was initiated by exposure of the strain to acetate sporulation medium, and monitored by the appearance of leucine-independent intragenic recombinants. Mutant isolation was based on the recovery of thermally induced defects in recombination. The temperature-sensitive characteristic was included to allow eventual characterizations of the temporal period during meiosis when each gene performs its essential function. Following mutagenesis with either ethyl methane sulfonate or nitrosoguanidine individual clones were tested at 34 degrees and 24 degrees for acetate-induced recombination. Starting with 2700 clones, derived from cells that survived mutagenic treatment, we isolated 48 strains with thermally induced lesions in recombination. In the majority of mutants premeiotic replication occurred normally, or nearly normally, at the restrictive temperature, indicating that the meiotic cycle was initiated and that there was a defect in an event required for intragenic recombination. We also detected mutants where the thermally induced lesion in recombination resulted from temperature-sensitive premeiotic DNA synthesis.


Antibiotics in biliary disease: the relative importance of antibiotic concentrations in the bile and serum. A randomised controlled trial has been performed in 150 patients undergoing biliary operations to determine whether an antibiotic which is excreted almost entirely in bile (rifamide) is to be preferred to one having satisfactory serum levels only (gentamicin). Patients were allocated to one of three groups: 50 received gentamicin, 50 received rifamide, and there were 50 controls who received no antibiotic cover. In the absence of duct obstruction, rifamide achieved extremely high bile levels but low serum concentrations. However, in jaundiced patients, both the bile and the serum concentrations of rifamide were too low to be of therapeutic value. The incidence of postoperative sepsis was not reduced by rifamide compared with controls. In contrast, gentamicin achieved adequate serum concentrations in 88% of patients. Despire poor bile levels, gentamicin was associated with a significant reduction of wound infection from 22% to 6% and septicaemia from 14% to 2% compared with controls. To reduce the septic complications of biliary operations, adequate serum levels of an effective antimicrobial are more important than an antibiotic, which is excreted almost entirely into the bile.


A double-blind comparison of sulpiride with chlordiazepoxide in neurosis. The therapeutic effectiveness of sulpiride on various types of neurosis was compared with that of chlordiazepoxide on a double-blind basis. Global improvement, effectiveness on various types of neurosis, and side effects were studied for a period of two weeks. The subjects consisted of 41 males and 32 females. Neurosis including borderline case and vegetative dystonia was divided into eight different subtypes comprising borderline, neurasthenic state, hypochondria, obsessive neurosis or phobia, depressive neurosis, anxiety neurosis, vegetative dystonia, and others. A newly devised matched-pair method of comparison was employed to achieve even distribution of the eight subtypes of neurosis between the two drug groups. As a result 40 patients fell in the sulpiride group and 33 patients in the chlordiazepoxide group. The backgrounds of the paired patients matched closely. The daily dose was uniformly 150 mg for sulpiride and 30 mg for chlordiazepoxide in the first week but was raised (or lowered in some cases) to 225 mg and 45 mg, respectively, in the second week according to severity. The rate of global improvement was 79% for the sulpiride group and 90% for chlordiazepoxide group, and the difference did not reach statistical significance. Improvement by manifestation (13 symptom items) and type of neurosis also matched. Side effects occurred at a rate of 28% (sulpiride group) and 30% (chlordiazepoxide group), and also matched closely in incidence and variety. The authors concluded that sulpiride in appropriate doses is useful in the treatment of neurosis without causing extrapyramidal side effects.


[Diagnosis of left-to-right shunt. Dye dilution curves in patients with left-to-right shunt during physical exertion and after beta-receptor blockade]. In 79 patients, suffering from central left-to-right shunts, dye-dilution curves were recorded partially using a peripheral venous injection site during reactive hyperaemia. 47 investigations were carried out during ergometer exercise in supine position. 32 patients received injections of 1-2 mg Propranolol or 0,4-0,8 mg Pindolol. During exercise dye-dilution curves showed characteristic alterations, indicating a decrease of the relative shunt-flow, whereas beta-receptor blocking agents increased the relative shunt-flow. Dye-dilution curves written during exercise may be helpful in evaluating the indication for operative closure in patients with small atrial or ventricular septal defects. beta-receptor blocking agents can facilitate diagnosis of left-to-right shunt as a screening test.


[Therapy of cervico-brachialgia. Controlled clinical comparison of a high-dose combination of neurotropic vitamins with an analgesic]. The study revealed that cervicobrachialgias (cervical syndrome) can be effectively treated with high-dosed neurotropic vitamins. This refers in particular to pain caused by pressure on the cervical plexus and paravertebral pain on pressur, Regarding paravertebral pain on pressure, it was possible to prove a significant superiority of the therapy with Neurotrat forte. The duration of therapy in both groups was, however, too short in order to prove a significant improvement regarding the lateral listing of the cervical spine. Both forms of treatment produced similar results in view of an improvement of impaired sensibility, paresthesia and alleviation of pain. In both groups side effects were only occasionally observed.


[Plastic surgery in the face and neck area. Use of regional transposition flaps]. Principles of fashioning of regional transposition flaps are described. The advantages of these flaps in reconstructing defects in the neck and head areas are explained using suitable illustrations.


[Physiological and pathological aging of the brain]. Also in the old age, the capability for a meaningful arrangement of life is dependent on the functional integrity of the brain. The processes of senile involution occurring physiologically do not exclude such a possibility. In normal aging, the reduction in the number of nerve cells and dendritic spines is small compared to pathological aging. There are no indications for transitions between normal and pathological aging. Senile dementia is an optionally occurring disease requiring an endogenous component for its manifestation. The psychic peculiarities occurring in the course of normal aging result mainly from environmental factors (i.e. psychic and social injuries). They need multidimentional treatment.


[Experiences with Epigard, a synthetic skin substitute, in the treatment of skin defects]. Epigard, a reticulated polyurethane foam laminated to a microporous polypropylene film, has been developed as a substitute for homograft and heterograft skin. After preliminary studies in animals, its clinical advantages and limitations were evaluated in 134 hospitilized patients with burn injuries and other skin defects. Examples for its indications are demonstrated and discussed. It is concluded that Epigard provides a satisfactory substitute for skin grafts with major advantages for ready availability, sterility and reduced cost.


[Functional-morphological kidney diagnosis in nuclear medicine]. Clinical indications and evaluation of basic nuclear medicine procedures (radionuclide-renography, rectilinear scanning) are discussed in regard to distinct diagnostic problems in urology and nephrology taking into account modern trends of their application. Additionally capabilities of special methods (sequential and functional scintigraphy, radionuclide-aortoangiography) are delineated including headlines of application from a clinician's point of view. Accuracy of the methods is presented based on a cooperational study of 17 nuclear medicine centers.


[Diagnostic strategy in diarrhea].

[Diagnostic strategy in diarrhea]. Planning the strategy in diagnosis of diarrhea some questions need to be answered: 1. Do the complaints in question fulfill the defined criteria of diarrhea? 2. Is it an acute diarrhea of infectious origin requiring fast microbiologic diagnosis? 3. Is it a chronic diarrhea? A) Do anamnestic findings and present status induce a certain suspected diagnosis, as in hyperthyreoidism, drugs, intestinal resections? B) Is a complete stepwise diagnostic planning needed as in Crohn's disease, insufficiency of the pancreas, non tropical sprue? Diagnostic measures have to be taken into consideration according to the answers given.


[Treatment of ulcus cruris. Clinical experiences using the Ulcrux cream and powder]. In an open trial 102 ulcera cruris of chronic venous genesis were treated locally with both Ulcrux Powder and Ointment in 54 female and 48 male patients over a period of 30 days. A positive result was obtained as to the shrinking of the wound area, the diminution of the substance defect, the development of granulations and the tendency towards epithelialisation. The influence on the cleansing of the wounds and on traumatic pain was also very favourable. Due to the rapid cleaning of the ulcera from necrotic films and to the good healing up tendency the therapeutic result was considered good to excellent with 81 patients (79,4%). Local side effects did not occur; the compatibility was also considered good in all cases. These test results prove that the combined treatment of chronic venous ulcera cruris with Ulcrux Ointment and Powder has real therapeutic advantages and seems to be superior to the traditional local forms of treatment.


[Analgesic effect of Neuro-Demoplas in a double-blind test]. Neuro-Demoplas was compared in a double blind trial on behalf of its analgesic properties to another antiphologistic containing high dosage of vitamin B complex and Metamizol. The onset of action of Neuro-Demoplas to inhibit pain was clearly more rapid, more intensive and more durable than with the other antiphlogistic.


[Amikacin in urinary tract infections with so-called problem pathogens]. Amikacin (Biklin) was applied in 33 patients with and without renal insufficiency, suffering of non-obstructive inflammations of the urinary tract, caused by multiple resistant hospital germs. The dosage was 125.0--1000.0 mg/die, injected as short infusion and solved in dextrose 5%. The serum and urine concentrations were measured simultaneously. In 18 cases an infection could not be seen after a control period of 8 days up to 4 weeks. Persistence of bacteria happened in five cases. Reinfection could be noted in 5 other cases. Side effects--as oto- or nephrotoxicity, pathological changes of the differential blood count, increasing of the serum transaminases and phosphatases--were seen neither during nor after the therapy.


[The history of breast feeding].

[The history of breast feeding]. The significance of breastfeeding, as considered millenniums ago, has been handed down by numerous contemporary reports and illustrations. The woman who suckles her baby has even since held an exceptional position in social life. In this essay the author describes the breastfeeding customs of far ancient people such as Egyptians, Greek, Romans and Indians. As a conclusion one might say that even today breastfeeding did not lose its importance for the relation between mother and child. It is an encouraging matter of fact that--unmindful of their emancipation--many "modern" mothers retain the breastfeeding habit. What would happen if breastfeeding was generally discredited, if mothers from all different countries would abandom breastfeeding in order to adopt bottle-feeding their children? Undoubtedly, that would not only cause a notable increase of the risks for the children's survival--which are due to many secondary factors--but, furthermore, it would mean an important expenditure for the budget for national economy which cannot be justified.


[Cervix insufficiency].

[Cervix insufficiency]. Between 1965 and 1975, Shirodkar operations were performed on 144 patients (mean age 28,6 years). The most frequent diagnosis was incompetence of the internal cervical os, which accounted for 90% of the operations, the remaining 10% being prophylactic. Sutures were performed between the 14th and 16th week of gestation and 72,5% of these patients had normal births (mean gestation time 39 weeks). Without Shirodkar operation, out of 291 pregnancies studied, 26% resulted in normal birth; whilst with Shirodkar, 82% of 137 pregnancies were successfully delivered at term.


[Clarification of cytologically doubtful findings of the cervix by DNA-single-cell-fluorescence-cytophotometry]. By using single-cell-DNS-cytophotometry in 28 single cases and 11 follow-up studies it was possible to differentiate the doubtful cytological findings and the mild and moderate dysplasia concerning the expected biological behaviour (risk of degeneration). If there are histograms similar to those with normal or inflammatory altered squamous epithelium, which are classified as "behaviour like a facultative precancerous lesion", further clinical observation is necessary. Histograms which show aneuploidy and/or wide spread to the right side are classified as "behaviour like an obligatory precancerous lesion". In these cases a conization should be performed, even if the findings are cytomorphological Pap III or mild dysplasias.


[Therapy of progressive chronic polyarthritis. Clinical studies of the effectiveness of azathioprine]. The present paper reports on the experience obtained in patients during treatment of rheumatoid arthritis with Azathioprine. The duration of treatment was on the average 16.5 months (minimum 1 month, maximum 54 months). The arthritic pains could be alleviated or even stopped completely in 73% of the cases. In 53.6% of the cases the morning stiffness was influenced favourably. The analgetic dose could be reduced in 54% of patients, who additionally received analgetics. Rheumatic attacks appeared rarely and also were of shorter duration and less severe. Therefore it can be concluded that Azathioprine has decreased the progredience of the disease. The main side-effects were gastrointestinal upsets. In 2 cases therapy had to be discontinued due to leucopenia. The results show that Azathioprine seems to be valuable for the treatment of severe rheumatoid arthritis. The overall effect can be considered as successful, but long-term therapy is necessary.


[Changes in hemostasis during epilespy treatment using dipropyl acetate. Extended study]. The influence of Dipropyl-Acetate (DPA, Ergenyl) on hemostasis parameters was examined in 12 patients. In 8 cases there was a prolongation of bleeding time, in 2 cases thrombocytopenia, in 6 out of 10 cases the platelet adhesiveness was pathologically reduced. We therefore recommend hemostasis-examinations--with emphasis on the thrombocytic system--in patients taking DPA.


[Effects and side effects of corticosteroids in allotransplantation]. The survival of rabbit skin allografts was significantly increased by local treatment of the grafts prior to transplantation. Triamcinolone acetonide proved to be the most effective compound from nine corticosteroids compared in this study. That its effect is mainly due to a local antiinflammatory activity could be clearly demonstrated after simultaneous transplantation of pretreated and untreated grafts on the same recipient and on pretreated second set grafts. In both experiments a significant prolongation of the survival of the pretreated grafts was observed. There were no side effects due to a general activity of the locally applied steroids. This was also confirmed by experiments with tritium-labelled Triamcinolone acetonide. By combining this local treatment with low dose general immunosuppression the same positive effect was seen without any side effects as with general treatment alone which, however, caused a high morbidity and mortality due to general side effects. The local pretreatment of kidney allografts with methylprednisolone did not increase the survival of the recipients in dogs. Comparative studies with general application of the steroids. Triamcinolone acetonide and methylprednisolone in rabbit skin allotransplantation revealed a superior immunosuppressive effect and a significantly lower risk of general side effects of Triamcinolone acetonide.


[Influence of intravenous steroid administration on kidney function]. The acute renal response to 1,0 g Prednisolon i.v. in kidney transplant recipients and normal controls was investigated. The results indicate an acute suppression of glomerular filtration rate (CIN, CCR) and effective plasma flow (CPAH). The reabsorption of sodium in the proximal tubule was shown to be impaired as a direct effect of the steroid infusion, as well as there was an increase in the filtered fraction of potassium. The creatinine clearance under treatment of high doses of steroids does not reflect the actual changes in glomerular filtration rate


[Isolation and characterization of seryl- and phenylalanyl-tRNA synthetase from yeast (author's transl)]. A procedure for the simultaneous isolation of seryl- and phenylalanyl-tRNA synthetase from yeast is described. In addition some other synthetases as well as tRNA nucleotidyltransferase can be obtained in an enriched state. The isolated seryl- and phenylalanyl-tRNA synthetases were compared to earlier preparations with respect to purity, specific activity, and structure. Previous investigations with fluorescence spectroscopy and kinetic methods were complemented and extended by experiments on the specificity of aminoacylation and on the isolation, by sucrose gradient centrifugation, of complexes between synthetase and tRNA or tRNA fragments. A protection of synthetases against inactivation by addition of substrates was observed. The dissociation of seryl-tRNA synthetase, at low concentrations, into monomer subunits was investigated by chemical modification with bifunctional reagents and by kinetic experiments. By modification of SH-groups fluorescent dyes were incorporated into both, seryl- and phenylalanyl-tRNA synthetase which retained most of their activity. The binding of tRNAPhe to phenylalanyl-tRNA synthetase which had been modified with pyrene maleimid was followed by fluorescence intensity measurements.


Multiple forms of lysyl-tRNA synthetase from Escherichia coli. Lysyl-tRNA synthetase has been isolated from E. coli. The enzymatic activity elutes as three or four bands from a hydroxyl-apatite column. Polyacrylamide gel analysis shows that each of these bands contain more than one enzymatically active protein species. The molecular weights of the subunits of these species provides an explanation for the variation in the molecular weights previously reported for this enzyme.


A computer package to facilitate compliance with utilization review requirements. The Cumberland County CMHC in Fayetteville, North Carolina, developed a series of computer programs to produce the information that is required by area regional utilization review committees. There is a total of eight programs. The first establishes a master file and prints out all the information that has been gathered on all the clients. The other seven programs retrieve various portions of the data stored in the master file. The system does not require elaborate in-house computer equipment. It is easy to operate and benefits clients by enabling clinical staff to spend less time on documentation and more time on direct treatment.


Membrane pathology of normal and malignant cells--a review. The plasma membrane has been shown to play a critical role in the regulation of cell growth and proliferation, and pathologic change in the plasma membrane has been detected in neoplastic cells. This article reviews the structure and functions of biological membranes and discusses the differences that have been reported in the plasma membranes of normal and malignant cells that may be associated with neoplastic transformation.


Chromosome preparations from microplate cultures of man, dog, rat, and mouse. A simple method for making chromosomal preparations from 10(5) leukocytes from man, dog, mouse, and rat and from 0.01 ml total human and dog blood is developed. The leukocytes and the peripheral blood are cultured in Cooke microtiter plates in a culture volume of 0.1 ml. The culture medium is R.P.M.I. 1640 supplemented with serum and phytohemagglutinin. The culture time is 72 or 96 h.


Human complementary component C'3: an appraisal. Results obtained so far on the C3 polymorphism suggest that the system should be a valuable marker in population studies. The instability of the complement component C3 may, however, cause some practical problems in population genetic fieldwork, since a certain fraction of serum samples may be difficult to type with certainity due to storage alterations. Studies of the conversion rate and the concentration of C3 have shown that there is no significant difference between the phenotypes. And furthermore, the fact that there is good agreement with the Hardy-Weinberg distribution indicates that the conversion has had no appreciable selective effect on the phenotype distribution (Brönnestam et al., 1971).


[Immunodiagnosis of rheumatoid arthritis author's transl)]. Rheumatoid arthritis (RA) presents a variety of immunological phenomena which can be of considerable diagnostic value. Of great significance is the determination of rheumatoid factors. Other autoantibodies are common but not specific for RA. Nevertheless the detection of such antibodies is beneficial for differential diagnostic considerations. Commonly demonstrated in rheumatoid arthritis are immune complexes which can be either soluble or within conclusions of circulating blood leucocytes or of synovial cells. Complement diminution can also be observed in RA, seldom in the serum, more often in synovial effusions. Like other humoral immune findings it lacks also disease specificity. A functional impairment of cellular immunity has also been postulated for rheumatoid arthritis, however the techniques for its demonstration are still cumbersome and cannot be easily used for routine diagnostic procedures. In conclusion, none of the immunological findings in RA is specific, however together with the anamnestic and clinical data it is possible to establish an exact diagnosis of this disease.


[Application of the immunofluorescent (antibody technique) in clinical immunology I. Technical Considerations (author's transl)]. The main problem of fluorescent antibody techniques is the elimination of "non-specific-staining (NSS)". To assure specificity of the reactions the following points have to be taken into account. The preparation of monospecific antisera, the coupling and purification of fluorescent conjugates to achieve optimal molar fluorescein-protein (F/P)-ratios and of appropriate specificity controls. Furthermore the testing of the optimal method for tissue preparation or the investigation of isolated cells is necessary. The paper shows that there is need for a standardization of the fluorescent antibody technique to achieve comparable results in different laboratories.


Effect on synthetic capacity of antibodies directed against receptors of allotype-bearing rabbit cells. Rabbit immunoglobulin molecules possess antigenically distinct allotypic markers. Immunoglobulin molecules possessing such markers which are attached to the surfaces of lymphocyte precursors of antibody-forming cells are believed to serve as antigen receptors. Interactions between allotypic determinants and antibodies directed against them can provide useful models for the study of immunoregulatory phenomena. Exposure of a fetal or neonatal rabbit to antibodies against one of its immunoglobulin allotypes results in a persistent defect in expression of that type (allotype suppression). Some evidence indicates that this phenomenon involves specific active suppression by lymphoid cells. By examining the effects of anti-allotype antibodies on allotype-bearing lymphocytes in vitro, certain specific events surrounding the processes of lymphocyte ontogeny, and the differentiation and maturation of antibody-forming cells are being analyzed.


Properties of the Fc receptor on macrophages. Macrophages and monocytes possess a surface receptor specific for the Fc region of certain subclasses of IgG. The binding site on IgG is localized within the Cgamma3 homology regions of the heavy chains. The intrinsic affinity of the receptor for IgG ranges from 10(6) to 10(8) M-1 depending on species and subclass of IgG. The most definitive studies on mouse macrophages indicate that IgG2a rapidly associates and dissociates from the receptor. The overall reaction is exothermic; increasing temperature lowers the intrinsic affinity. The Fc receptor, in common with many other membrane components, may be capped by polyvalent ligands under permissive conditions and capping is inhibited by azide. Data on the chemistry of the receptor is both sparse and conflicting. Sensitivity of the receptor of proteolytic enzymes has been clearly demonstrated for mouse macrophages although rabbit and guinea-pig cells appeared to carry resistant receptors. IgG-binding may be inhibited if cells are treated with phospholipases and certain group-specific reagents. Evidence is reviewed indicating that the Fc receptors found on various cell types are different. The macrophage Fc receptor appears to play a role in mediating phagocytosis and in non-immune cytotoxicity. Whether the receptor serves only to concentrates sensitized target cells at the cell surface or whether occupation of the receptors results in modulation of effector cell function remains to be determined.


Evolutionary and developmental aspects of T-cell recognition. Studies relating to the nature of the antigen-specific T-cell receptor are reviewed in the light of present knowledge of phylogenetic and ontogenetic development. It is suggested that this evidence supports the concept that immunoglobulin (Ig) is the T-cell receptor, and that the following conclusions may be tentatively drawn.


The immunology of the insulin receptor.

The immunology of the insulin receptor. We have detected and characterized anti-insulin-receptor autoantibodies which circulate in several patients with insulin resistance diabetes. These antibodies are predominantly IgG and are polyclonal. They inhibit insulin binding to its receptor on a variety of tissues from widely separated species. Antibodies obtained from different patients appear to bind to different determinants on the receptor and alter receptor function in several ways. Some anti-receptor antibodies are capable of stimulating insulin-like effects on target tissues, while others block insulin-stimulated effects. Direct labeling of anti-receptor antibody with 125I permits use of these antibodies as an assay and probe of insulin receptors.


Immunological probes into the mechanism of cholera toxin action. The use of antibodies to specific cell surface proteins or to ligands which interact with cell surface receptors is a powerful tool for analyzing the properties of membrane proteins and the consequences of specific cell surface ligand-receptor interactions. Two central observations concerning membrane structure and function, - the diffusibility of membrane proteins (1) and ligand-triggered modulation of specific receptors (2), have derived from the use of antibodies to analyze the properties of membrane proteins. In our study of the mechanism of action of cholera toxin, a protein which binds to a specific cell surface receptor and results in the activation of adenyl cyclase, considerable information has been gained through the use of immunological techniques. This review will briefly summarize the data underlying our current concept of cholera toxin action at the cell membrane and will emphasize those observations made through the use of immunological approaches.


Some physical chemical aspects of receptor-ligand interactions. Quantitative evaluations of a variety of binding reactions of interest to immunologists have usually assumed that the reactants are homogenously dispersed in solution for purposes of calculation. In fact, many of these reactions involve cell-bound "receptors" which are present at exceedingly high local concentrations on discrete particles. We describe how such a distribution can influence the experimentally measured binding constants. We also briefly consider the additional complexities introduced when multipoint attachment between the ligand and cell-bound receptors is possible, and discuss the possible biological implications.


Antisera to mast cells and the receptor for IgE. Recent studies on the receptor for IgE on rat mast cells and rat basophilic leukemia cells have established that this receptor has a molecular weight of about 60,000 daltons and is, at least in part, a protein. However, so far no antisera, monospecific for this receptor, are available. Only antisera to mast cells have been described. Such antisera are capable of releasing histamine from mast cells, but only in the presence of complement. In most instances such antisera were not specific for mast cells only, but in some cases they were rendered specific by absorption with lymph node cells or liver cells. One report demonstrates that such an absorbed antiserum was capable of inhibiting the binding of IgE to mast cells. Moreover, it precipitated several mast cell surface components. One of these possibly was the receptor for IgE.


Inhibition of B lymphocyte activation by interaction with Fc receptors. Much early work indicated that specific antibody can play an inhibitory role in the immune response. This inhibitory activity was found to be dependent on an intact Fc portion of the antibody used, both in vivo and in vitro. Thus a role for lymphocyte Fc receptors in regulations of the immune response was suggested. However, soluble antigen-antibody complexes, which bind to Fc receptors, do not appear to inhibit B cell activation. Recent experiments have demonstrated that antigen-antibody complexes immobilized on a surface strikingly inhibit LPS induced B cell mitogenesis and polyclonal antibody synthesis. Mechanisms for Fc receptor-mediated inhibition of B cell activation have been considered, and a model proposed to explain many of these findings as well as allow for antigen specific inhibition.


Characterization of the virulence and antigenic structure of Salmonella typhimurium strains with lipopolysaccharide core defects. The virulence and antigenic characters of Salmonella typhimurium strains, identical except for known lipopolysaccharide core defects, were compared. Smooth strains multiplied extensively and killed most mice. Deep rough strains containing only heptose I or heptose I and II in the rough core were completely eliminated after 6 h, whereas more superficial rough strains containing additional core sugars could be detected in low numbers (10(4) colony-forming units/g of tissue) for at least 7 days postinjection. Normal human serum exhaustively absorbed with certain rough strains was tested for ability to kill other rough strains. Two strains with the most superficial defects (rfaJ, rfaL) each had a unique serological character; strains with deeper defects showed much cross-reactivity. Similarities between the susceptibility of strains to the bactericidal effect of specifically absorbed serum correlated, in some cases, with similarities in in vivo behavior.


Immunogenicity of a purified and carrier-complexed streptococcal lipoteichoic acid. Solublie teichoic acid could not stimulate the formation of antibodies either as an aqueous solution or in an emulsion with Freund incomplete adjuvant. However, when given as precipitates with methylated bovine serum albumin, it stimulated strong anti-teichoic acid responses. High levels of anti-teichoic acid antibodies usually resulted in allergic symptoms upon secondary challenge either with immunogenic teichoic acid-methylated bovine albumin complexes or with non-immunogenic soluble teichoic acid. These symptoms suggested anaphylatic shock, but probably also included the formation of complement-consuming immune complexes.


Role of platelets in the pathogenesis of canine endotoxin shock. Endotoxin-platelet interactions are thought to be of major importance in the response of dogs and other species to bacterial endotoxin; the mechanisms postulated are: (i) the release of vasoactive substances, (ii) the formation of occlusive platelet aggregates, and (iii) induction of intravascular coagulation. The role of platelets in canine endotoxin shock was examined in animals with thrombocytopenia induced by estrogen pretreatment (less than 10,000 platelets/mm3) and in controls. After intravenously administered endotoxin, the hemodynamic responses, mortality, and gross necropsy findings were similar in both groups. These data indicate that endotoxin-platelet interactions are not determinative in the pathogenesis of canine endotoxin shock.


Effect of enterobacterial common antigen on mouse virulence of Salmonella typhimurium. A series of nearly isogenic O4-12, and O-6,7 Salmonella typhimurium strains differing in regard to the enterobacterial common antigen (ECA) were constructed by conjugation. When tested in intraperitoneal infection of mice, the O-4,12 strains containing ECA were more virulent than their O-4,12 sister recombinants without ECA (P = less than 0.001). The same difference could be shown with ECA-positive and ECA-negative S. typhimurium derivatives, whose O antigens were of the group C type (O-6,7). The ECA-positive and ECA-negative O-4,12 strains did not differ in their growth rates in broth or clearance rates in vivo.


Treponema pallidum infection in subcutaneous polyethylene chambers in rabbits. Male New Zealand white rabbits with subcutaneous polyethylene chambers in place for at least 3 months were inoculated by one of the following three methods: (i) "intra-chamber" (IC) inoculation with "normal" chamber fluid: (ii) intratesticular inoculation with Treponema pallidum; or (iii) IC inoculation with T. pallidum. Rabbits given dexamethasone only, oxisuran only, both drugs, or no drug were observed serially after inoculation. T. pallidum survived and temporarily multiplied to significant numbers within subucutaneous chambers after IC inoculation in rabbits given dexamethasone. In rabbits not treated with dexamethasone, T. pallidum counts in chamber fluid decreased rapidly and remained at low levels for 30 days after IC inoculation. Oxisuran appeared to have little or no effect on T. pallidum multiplication. All rabbits studied had a nonreactive serum and chamber fluid serological test for syphilis before inoculation. All rabbits inoculated with T. pallidum eventually developed reactive serum and chamber fluid serological tests. The IC route of inoculation was associated with a delay in the development of serum serological reactivity and with earlier chamber fluid reactivity as compared with the intratesticular route of inoculation. An immediate but transent influx of polymorphonuclear leukocytes was associated with IC inoculation of T. pallidum. Chamber fluid total protein content declined very slightly in all groups of rabbits during the month after inoculations. Successful cultivation of T. pallidum in an in vivo setting suggests that this animal model may be useful in further studies of the biology of the organism of the pathogenesis, immunology, and treatment of syphilis.


Distribution of dengue-2 antigens by electron immunocytochemistry. The distribution of dengue-2 antigens was studied in infected monkey kidney cells (LLC MK2) using an indirect, horseradish peroxidase-conjugated immunoglobulin technique. This procedure allowed both light and electron microscopic examination of serial-step sections of individual cells cut in a plane perpendicular to the monolayer. Both virion and nonvirion antigens were identified on the plasma membrane with this technique. A diffuse cytoplasmic reaction product was also present. The intensity and distribution of the cytoplasmic reaction product was related to disruption of the plasma membrane.


Partial purification and characterization of a heat-labile enterotoxin of Escherichia coli. A partially purified enterotoxin was obtained from the growth medium of Escherichia coli strain 711 (P307), a derivative of E. coli K-12, by ultrafiltration, precipitation with ammonium sulfate, molecular sieving, and anion exchange column chromatography. The active moiety, which is heat-labile, behaved like a protein particle of 180,000 to 200,000 daltons during molecular sieving and ultracentrifugation. During polyacrylamide gel electrophoresis in sodium dodecyl sulfate (SDS-PAGE), it dissociated into two subunits with apparent molecular weights of 68,000 to 70,000 and 14,000 to 15,000. SDS-PAGE after heating in SDS changed the larger subunit to an apparent molecular weight of about 40,000; the smaller subunit did not change. The intact particle induced rounding of the cells in Y-1 mouse adrenal tumor cells used for assay. The detergent-dissociated molecules were not active. Proteolysis of the purified toxin by tolylsulfonyl phenylalanyl chloromethyl ketone-trypsin appeared to enhance its activity. The addition of serum to the assay medium resulted in partial depression of the activity. Activity was also abolished by preincubation of the toxin with either a rabbit antiserum to it or solutions containing GM1 ganglioside. The length of time needed to evoke a response in the assay system by fractions from different stages in the purification of the enterotoxin was a useful parameter in the evaluation of specific activity.


Gastrointestinal candidiasis in rats treated with antibiotics, cortisone, and azathioprine. Conventional albino rats treated with peroral chloramphenicol, gentamicin, and/or parenteral cortisone were challenged with Candida albicans Antibiotics and cortisone were equally effective in predisposing the animals to colonization by the fungus. All animals treated with both antibiotics and cortisone developed defined, focal, superficial invasion of the cornified squamous epithelium of the stomach next to its junction with the glandular mucosa, as well as focal superficial invasion of the esophagus. Equivalent yeast cell and mycelial inocula of C. albicans were equally effective in producing colonization and invasion of the gut. Dissemination of the fungus from the gut was not found even after the addition of azathioprine to the treatment regimen; however, such addition did predispose to more extensive and severe lesions of the esophagus and stomach. Approximately 25% of infected cortisone- and antibiotic-treated rats developed agglutinins against C. albicans by 22 to 23 days after challenge, whereas 15% developed precipitins. The antibiotic-cortisone-treated rat may be a useful and consistent experimental model in the study of gastrointestinal candidiasis.


Bacterial competition as a means of preventing neonatal diarrhea in pigs. Baby pigs orally inoculated with a porcine strain of enterotoxigenic Escherichia coli (K88+,Ent+) showed signs of depression, severe diarrhea, and, in some instances, death. Few, if any, signs of illness occurred if baby pigs were first inoculated with a K88 possessing non-enterotoxin-producing strain of E. coli.


Plasmid-mediated properties of a heat-stable enterotoxin-producing Escherichia coli associated with infantile diarrhea. The plasmid mediation and transmissibility of heat-stable enterotoxin production and multiple antibiotic resistance have been demonstrated for Escherichia coli O78:K80:H12 epidemiologically incriminated in a hospital outbreak of infantile diarrhea. The conjugal transfer of a 67 X 10(6) - and a 30 X 10(6)-dalton plasmid was associated with the transfer of resistances and enterotoxin production, respectively. Using antibiotics to select E. coli K-12 transconjugants from a one-step bacterial cross, all of the monitored resistances were transferred concurrently, and 36% of the resistant transconjugants produced enterotoxin.


Depression of the immune response to sheep erythrocytes in mice infected with Taenia crassiceps larvae. Intraperitoneal infection of mice with larvae of the cestode parasite Taenia crassiceps results in depression of both primary and secondary antibody responses to sheep erythrocytes in vivo. The depression is not associated with a shift in kinetics of the response. Both immunoglobulin M (IgM) and IgG responses are depressed, but IgG is depressed more than IgM in the secondary response. Secondary in vitro sheep erythrocyte responses are consistently depressed in both spleen and mesenteric lymph node cell preparations from infected mice, whereas primary in vitro sheep erythrocyte responses are consistently depressed in mesenteric lymph node cell preparations but not always in spleen cell preparations. These results are consistent with antigenic competition. The cell type or types involved in mediation of the immunological defect in the infected animals remain to be identified.


Influence of rodent malaria on the course of Leishmania enriettii infection of hamsters. Plasmodium yoelii infection was established in hamsters, and the effect of this type of malaria on concurrent Leishmania enriettii infection was examined. It was found that the course of the L. enriettii infection was affected by P. yoelii and that this effect depended on the relative timing of the two infections. A chronic malarial infection with Plasmodium berghei was also established in hamsters, and this was found to affect the course of a concurrent L. enriettii infection in a similar manner to P. yoelii. These results are discussed in relation to current knowledge of the immunosuppressive effects of plasmodia.


Formation of crystalline deposits by several genera of the family Enterobacteriaceae. Several species of bacteria from the family Enterobacteriaceae formed crystalline materials containing calcium when grown in a defined culture medium. Enterobacter aerogenes, Proteus vulgaris, Citrobacter freundii, and C. intermedius produced calcium pyrophosphate crystals. Edwardsiella tarda and Escherichia coli formed calcite III crystals, whereas Proteus mirabilis, Klebsiella pneumoniae, Providencia stuartii, and Serratia marcescens produced hydroxyapatite crystals. Several of these bacteria have been isolated from the kidneys of patients with kidney stones, indicating that microorganisms may be involved in the enucleation process of kidney stone formation.


Phage conversion to hemagglutinin production in Clostridium botulinum types C and D. Five toxigenic strains of Clostridium botulinum types C and D were incubated at 37 degrees C for 7 days in 15 ml of the following media: LYG medium, cooked-meat medium, egg meat medium, and N-Z-amine medium. The supernatants of these cultures were tested for hemagglutinin production with 1% erythrocytes obtained from mice, guinea pigs, chickens, sheep, monkeys, and humans. Four toxigenic strains produced hemagglutinin. The highest hemagglutinin titer was obtained with a combination of human erythrocytes and cultures incubated in LYG medium. When the same experiment was carried out with many nontoxigenic strains, hemagglutination was observed in only one strain, C-N71. Strains producing hemagglutinin also produced phages. The phages obtained from toxin- and hemagglutinin-producing strains converted nontoxigenic indicator strains to produce both toxin and hemagglutinin. The phage obtained from a toxin-positive hemagglutinin-negative strain could only induce cultures to produce toxin, and the phage from a toxin-negative hemagglutinin-positive strain could only induce production of hemagglutinin. These studies suggest that the production of hemagglutinin by C. botulinum types C and D is governed by bacteriophages and that hemagglutinin production can be transmitted separately or concomitantly with toxin production.


Variability of protection in inbred mice induced by a ribosomal vaccine prepared from Salmonella typhimurium. Ribosomal vaccines prepared from Salmonella typhimurium were effective immunogens in A/J inbred mice and C3H/HeTex, inbred mice. However, ribosomal vaccines were not protective in C57BL/6J inbred mice. A/J mice were protected against lethal challenge by attenuated S. typhimurium live-cell, ribosomal, phenol, and heat-killed vaccines. C3H/HeTex mice were protected by live-cell, ribosomal, and phenol vaccines but not the heat-killed vaccine. Only the live-cell vaccine gave significant protection in the C57BL/6J inbred mice. A comparison of the kinetics of infection in sham-immunized mice and mice immunized with ribosomes showed that ribosome preparations elicited protection against Salmonella infection in mice inherently sensitive and resistant to Salmonella.


Isolation of skin permeability factors from culture filtrates of Salmonella typhimurium. Engerotoxins isolated from Vibrio cholerae and toxigenic Escherichia coli cause permeability alterations in rabbit skin. Firm induration and erythema are observed within 18 to 24 h, and visualization of the reaction may be enhanced by intravenous injection of Pontamine sky blue dye. Two skin permeability factors (PF) have been found in culture filtrates of Salmonella typhimurium. A rapid acting factor, produced optimally in brain heart infusion broth at 37 degrees C by numerous Salmonella species, has a critical bluing time of 1 h after completion of skin testing. This rapid PF is heat stable at 100 degrees C for at least 4 h and has no associated induration. The delayed factor is heat labile, being completly destroyed within 30 min at 75 degrees C and causes marked duration of the rabbit skin within 18 h that is indistinguishable from the permeability reactions of V. cholerae and E. coli enterotoxins. Induration produced by the delayed PF is observed only after chromatography of the culture filtrate on a Sephadex G-100 column. Thus, the effects of the delayed PF appear to be masked or blocked by an inhibitor-like substance present in crude culture filtrates. Both early and delayed factors are estimated to have a molecular weight of at least 90,000. It is postulated that one or both of these factors may participate in the pathogenesis of Salmonella infections.


Nature of the effector cells responsible for antibody-dependent cell-mediated killing of Cryptococcus neoformans. Studies were performed to identify the types of human peripheral blood leukocytes capable of killing Cryptococcus neoformans in the presence of anticryptococcal antibody in vitro. A total of 24.1 +/- 2.7% (mean +/- standard error of the mean of four experiments) of the original cryptococcal inoculum survived in a mixed mononuclear, cell preparation (approximately 30% monocytes) after 4 h of incubation at 37 degrees C with rabbit anticryptococcal antibody. When phagocytic cells were removed, there was 36.4 +/- 4.6% survival in six experiments, compared with 52.8% survival in the presence of purified granulocytes (mean of two experiments) and 96.9 +/- 1% survival in the presence of purified T cells. There was never any significant killing in control mixtures that contained leukocytes with normal rabbit serum nor in those that contained anticryptococcal antibody without effector leukocytes. Significant antibody-dependent fungicidal activity was seen with ratios of effector to target cells as low as 6.25:1. These observations indicate that multiple types of peripheral blood leukocytes, excluding T cells, are capable of antibody-dependent fungicidal activity.


Neonatally thymectomized Lewis rats infected with Mycobacterium leprae: response to primary infection, secondary challenge, and large inocula. Several experiments were carried out to measure the ability of neonatally thymectomized Lewis rats (NTLR) to limit multiplication of Mycobacterium leprae. NTLR inoculated in one hind footpad with 10(7) viable M. leprae and challenged in the other hind footpad with 5 x 10(3) organisms simultaneously or 120 or 180 days later permitted multiplication in both sites. By contrast, immunologically intact rats similarly inoculated did not permit multiplication from either inoculum. NTLR and immunologically normal BALB/c mice were equally susceptible to infection with M. leprae, in that multiplication occurred regularly in the footpads of both species when inoculated with a bacterial suspension diluted to provide five organisms per footpad. Finally, multiplication occurred when five viable M. leprae diluted with 10(7) heat-killed organisms were inoculated into the footpads of NTLR. Although there was some evidence that NTLR are not completely immunosuppressed, NTLR appear to be capable of detecting much smaller proportions of viable M. leprae than can be detected by immunologically normal mice.


Further purification of group A streptococcal pyrogenic exotoxin and characterization of the purified toxin. Streptococcal pyrogenic exotoxin (SPE) isolated from culture filtrates of strain NY-5 (type 10), and separated from other extracellular by differential solubility in ethanol and acetate-buffered saline, has previously been shown to exhibit a wide range of biological activities including erythrogenic activity, pyrogenicity, enhancement of susceptibility to endotoxin shock, blockage of the reticuloendothelial system immmunosuppression, and lymphocyte mitogenicity. Toxin prepared in this way was found to consist of hyaluronic acid and several proteins which could be distinguished by thin-layer polyacrylamide isoelectric focusing (IEF), SPE has been further purified by ion exchange chromatography on QAE-Sephadex columns. One of the fractions isolated from QAE-Sephadex, and shown to be a homogenous protein by thin-layer IEF and Ouchterlony with hyperimmune serum, was highly active erythrogenically, pyrogenically, and in enhancing susceptibility to endotoxin. This fraction was identified as exotoxin A. A second, less active fraction identified as SPE B showed similar activities, but differed from the other fraction antigenically and in net charge and molecular weight. These findings indicate that a single highly purified protein can mediate at least three of the biological activities attributed to SPE and NY-5 produces pyrogenic exotoxins A and B in vitro as well as in vivo.


Radioimmunofluorescent antibody technique for detection of reovirus antigen in cell culture. Antibody against reovirus type I must partially purified and conjugated with fluorescein isothiocyanate (FITC). The FITC-labeled antibody was then conjugated with 125I. A gamma globulin fraction of normal sera was similarly labeled with FITC followed by a 131I label. The FITC + 125I-labeled immune reagent was then mixed on an equal protein basis with the FITC + 131I control reagent. This mixture was used to stain acetone-fixed reovirus-infected cover slips. After staining, the cover slips were examined by fluorescence microscopy. Infected cover slips demonstrated characteristic reovirus immunofluorescence, whereas uninfected cover slips were negative. After visual examination, the cover slips were placed in tubes and counted in a two-channel gamma analyzer. By comparing the quantitative isotope data with the qualtitative information from immunofluorescence on a single preparation, it was possible to correlate antigen production sites with quantitative production values.