[Karl Kraus and psychiatry. An assay].

This essay deals with Karl Kraus' polemics against psychiatry; not only orthodox psychiatry but also psychoanalysis, which at that time was gaining in importance. The introduction is an appreciation of Kraus' significance for the German speaking civilization. Then the two cases, Louis von Sachsen and Prof. Beer, are discussed in relationship to Kraus' polemics against forensic psychiatry, for in these cases  forensic psychiatry was in danger of losing its claim of being scientific and of becoming a supporter of the vested interests or an advocate of sexual repression. It is then attempted to show how aphorisms with universal validity arise out of topical polemics. Kraus' position in relation to Freud and his school is discussed. We try to demonstrate that the similarity in opinion that H. Marcuse assumed about the significance of a sexual economy can only be a superficial one, especially as it relates to sublimation as a mainspring of civilization. Also Kraus' critique of the inferiority complex is delved into. Concluding, we touch upon attempts at interpreting Kraus' work and personality from a psychoanalytical standpoint. Here the term 'narcissistic personality' is discussed.


Bacterial infection in relation to pain and irregular bleeding in women using IUD.

Pain and metrorrhagia in women using IUDs are often the reason for removing the IUD. In 54 women where the IUD was removed, uterine fluid for bacteriologic cultures was taken from the removed IUD. There was a correlation between positive cultures and symptoms of pain and metrorrhagia. Spotty bleeding caused by microtraumata and increased fibrinolytic activity of the IUD might thus consitute a substrate for bacterial invasion resulting in endometritis and in turn increased bleeding and pain. In 18 male consorts of females with positive cultures, asymptomatic bacteriospermia was found. Treatment offered alleviated symptoms. Removal of the IUD need thus not be necessary.


Neural and chemical control of abomasal secretion in sheep.

Abomasal secretion has been collected from Grosskopftype pouches prepared in five adult Karrakul ewes. Feeding caused a marked increase in volume, acidity and pepsin activity, whereas these factors were reduced by starvation. Both insulin-induced hypoglycaemia and electrical stimulation of the vagal nerve supply to the pouch increased the volume, total acid and pepsin secretion. It is concluded that vagal activity is an important factor in the control of abomasal secretion in sheep.


Ventilator modifications for intermittent mandatory ventilation.

A Loosko MK2 ventilator has been modified to provide IMV in newborns. IMV rate can be varied from 3-60/min. The minimum inspiration period can be theoretically as low as 0.1 sec. This modification in neonatal mechanical ventilation has been shown to be economically feasible.


Cardiac blood pool imaging: a review.

The field of cardiac blood pool imaging has been reviewed. The radioisotopes and radiopharmaceuticals used for this purpose have been discussed. Data related to the radiation exposure from these agents were presented and compared with the radiation exposure of a conventional contrast angiogram. Application of cardiac blood pool imaging to the qualitative evaluation of pericardial effusions, cogenital heart disease, acquired heart disease, and cardiac shunts have been discussed. Quantitative methods for measuring cardiac output, stroke volume, left ventricle ejection fraction, left ventricle end-diastolic volume, percent mitral valve regurgitation, and Qp:Qs flow ratios in both right-to-left and left-to-right shunts were presented in detail. Where possible, correlation with contrast angiographic findings was given. Newly developed techniques of ECG gating of cardiac images were also described in detail. This description has included applications of these techniques for evaluation of regions of myocardial dyskinesis and to quantitation of left ventricle diastolic volume, ejection fraction, time between ejection onset and peak systolic flow, ejection duration, and peak circumferential fiber shortening. Once again, these were correlated with contrast angiography where possible. It is concluded that cardiac blood pool imaging is a useful procedure capable of yielding good qualitative and quantitative results in a wide variety of clinical cardiac diseases and that these results compare favorably with those obtained by contrast angiography. Although there are limitations in the present techniques, solutions to at least some of these limitations are possible. Because of its simplicity, safety, and low cost relative to cardiac catheterization, more widespread use of these procedures is urged.


The differential diagnosis of the unilateral small kidney.

Although it is frequently possible to determine the cause of an unilateral small kidney, there are cases where the renal damage is so advanced, or the interference to growth and maturation so extensive, that the etiology cannot be established. High-dose urography with tomography during the nephrographic phase are required for a confident diagnosis. Ureteral compression should be used to improve calyceal visualization. This technique can be adapted for children. Complementary diagnostic procedures are often needed, both in adults and in children. We cannot emphasize enough the need for clinical correlation and comparision with previous examinations.


Current status of mammography.

Great progress has been made in recent years in the diagnosis and treatment of breast cancer; however, breast cancer continues to be the most common and lethal cancer in women today. Early diagnosis is essential in order for treatment to be given before the tumor spreads beyond the breast. The radiation risks of mammography have been greatly reduced with the use of newer low-dose techniques, and the benefits of mammography have increased because of a better understanding of the natural history of breast cancer, as well as improved methods of treatment. Radiologists must continue to take an active role in the early diagnosis of breast cancer. Continued research is needed in order to improve screening methods and to develop newer, noninvasive techniques.


The characterization and functional significance of plasma membrane Fc Receptors.

The importance of various components of the plasma membrane in the generation of the immune response has long been recognized. The identification and characterization of functional membrane receptors are essential for an understanding of the molecular basis of the immune response. The Fc receptor can be operationally defined as a site on the plasma membrane which is capable of binding the Fc portion of IgG. The Fc receptor has been reported on both lymphoid and nonlymphoid cell types. The functional significance of the receptor on these cell populations, as well as the biochemistry of the receptor, is not well established and is currently an important area of investigation. In this review, the Fc receptor will be discussed, primarily in human and murine lymphoid and macrophage cell populations, as a model of membrane immunoreceptors. The review will be divided into four sections. In the first section, consideration will be given to the binding assays used to study Fc receptor-ligand interactions and the class and subclass specificity of the receptor. The possibility of receptor-receptor interaction and quantitation of receptor sites will be discussed. The second phase of the review will be concerned with the fractionation and biochemistry of this receptor system, emphasizing the differences reported in receptor composition from different cell sources. In the third section of the review, the biologic significance of the receptor will be examined. The importance of the Fc receptor in opsonization and phagocytosis by mononuclear phagogytes, B-cell activation, antibody-dependent cell cytotoxicity, and the possible relationship of Fc receptors to the histocompatibility-linked immune response genes will be discussed. Finally, in the fourth section, the possible mechanisms of receptor regulation will be considered. Cell fusion experiments which have been performed and the expression of receptor activity on the hybrid cell clones will be discussed. Receptor regulation at the physiologic level will include the effects of cyclic nucleotide levels and cell cycle on receptor expression. It is the intent of this review to provide a detailed analysis of the Fc receptor and to emphasize its importance in microbiology, cell biology, and immunology.


Cellular antimicrobial immunity.

Acquired resistance to infectious disease may be expressed by a predominantly humoral or a cellular mechanism or, more frequently, by a combination of the two. The cellular interactions which are responsible for the induction of the immune response in the skin, lung, intestinal mucosa, genitourinary tract, conjunctiva, and peritoneal cavity are discussed and the role of living or dead vaccines in the induction of acquired resistance is outlined. The host response involves three different cell types: the phagocytic cell (polymorphs or macrophages), the thymus-dependent (T) lymphocyte, and the thymus-independent (B) lymphocyte-plasma cell line. The normal unstimulated phagocytic cell is capable of killing most nonpathogenic bacteria that gain entry to the tissues. However, the presence of opsonic antibodies and activated macrophages is required to eliminate the pathogenic intracellular parasites. Such immunological activation involves the presence of sensitized T-lymphocytes in the lesion. The cellular response is also characterized by the simultaneous development of a state of delayed-type hypersensitivity (DTH), along with the antimicrobial CMI response. A rising humoral response normally develops subsequently. Killed bacterial cells (except when incorporated into Freund's complete adjuvant) induce the humoral response without the CMI reaction so that such vaccines are not able to fully protect the host against the naturally acquired disease. With the development of cell fractionation methods as well as the identification of distinctive cell surface markers, suspensions of B- and T-cells and macrophages can now be prepared for use in increasingly sophisticated transfer and reconstitution studies. The role of the different cell types in the expression of humoral and cellular immunity has been determined, and the effect of various immunopotentiating and immunosuppressive regimens on the immune system as a whole has been evaluated quantitatively. These studies have led to an appreciation of the role played by suppressor B- and T-cells in the interplay of both humoral and cellular components of the host defense system during the development of immune tolerance, desensitization, anergy, autoimmunity, and the expression of an anamnestic immune response following reinfection.


The buried horizontal mattress suture.

The buried horizontal mattress suture (BHMS) is a new tension suture technique using synthetic absorbable polyglactin 910 suture. When used in minor surgery of the trunk and extremities, this technique eliminates significant suture marking.


Pigmentary correction of piebaldism by autografts. II. Pathomechanism and pigment spread in piebaldism.

Human piebaldism is an expression of neural crest component abnormality that is classically inherited dominantly and is probably heterogeneous genetically. Though usually a stable cutaneous condition, spontaneous pigmentation in hypomelanotic tracts is known. Here, pigmented skin around the axilla was transplanted to hypomelanotic areas in two patients with piebaldism. The cutaneous autografts, whether full-thickness punch grafts, split-thickness shave grafts, or epidermal suction grafts, retained their capacity of pigmentation. Moreover, there was spread of pigmentation around the grafts. Strategic placement of cutaneous autografts in the most conspicuous hypomelanotic areas of piebald persons can improve their appearance.


Intraoral blue nevus--a review.

The clinical characteristics and appearance of the intraoral blue nevus are reviewed herein. The histogenesis, histologic appearance, differential diagnosis, and treatment of this uncommon oral lesion are also discussed.


The routine use of positive end-expiratory pressure after open heart surgery.

Because atelectasis of the left lower lobe is a frequent complication of open heart surgery, we evaluated the efficacy of routine therapy with positive end-expiratory pressure (PEEP) to prevent this complication. Twenty-four patients were randomly assigned to either a group receiving therapy with PEEP (ten patients) or to a group with no PEEP (14 patients). The two groups could not be distinguished by age, weight, the forced expiratory volume in one second (FEV1), the ratio of FEV1 over the forced vital capacity, the time on the pump, the units of blood transfused, the tidal volume, or the hours of mechanical ventilation. There was no significant roentgenographic difference between the two groups in either the degree or frequency of left lower lobe atelectasis. While the arterial-alveolar ratios tended to improve over time in those patients receiving therapy with PEEP, this improvement was not clinically significant. No complications were encountered with the use of PEEP. We conclude that the routine use of PEEP following open heart surgery is safe but offers no advantage over standard ventilatory techniques.


Combined action of sulphamethoxazole and trimethoprim against clinically-isolated sulphonamide-resistant bacteria.

42 strains of gram-negative bacteria, isolated from clinical material, resistant to trimethoprim (TM), sulphamethoxazole (SMZ) or both, have been tested for synergistic sensitivity to TM + SMZ. The combination showed no synergy against strains 'highly resistant' to SMZ (MIC greater than 1 mg/ml). Indifference was invariably observed. However, synergy was seen with strains 'moderately resistant' to SMZ (MIC 100--1,000 microgram/ml). Our results indicate that present techniques for the testing of urinary tract isolates for sensitivity to co-trimoxazole need revision.


Cyclacillin: in vitro activity against aerobic and anaerobic bacteria.

Cyclacillin at 50 microgram/ml inhibited streptococci (100%), shigella (72%), and Bacteroides (B) fragilis ss fragilis (84%). At 100 microgram/ml it inhibited proteus (75%), B. fragilis (86%), and E. coli (50%). Cyclacillin was more effective than ampicillin against beta-lactamase-producing H. influenzae. The activity of cyclacillin was found to be more enhanced when tested in nutrient agar and Mueller-Hinton agar than in brain heart infusion agar.


Advances and disappointments, indications and restrictions regarding HLA-matched corneal grafts in high risk cases.

54 corneas, 49 high risk cases and 5 low risk cases, were grafted with HLA-matched donor material. The importance of antigen compatibility especially in high risk cases is demonstrated. The percentage of irreversible rejections and reversible rejections in this group is markedly smaller as compared with our own "at random" material and the "at random" material of other authors. The follow-up period is 3--21 months.


Diagnosis and treatment of complications in the follow-up period after corneal transplantation.

Corneal transplantations are usually performed in Eye Clinics. The patient's own ophthalmologist usually performs the follow-up examinations because the patients often live a long way from the Clinic. It is very important that these ophthalmologists can diagnose the complications which may occur in the follow-up period and know how to treat them. The complications considered are: glaucoma, marginal infiltrations, epithelial defects suture infections, wound dehiscence, disturbances of the tear-film and recurrent herpes. Special mention is made of the rejection reaction. This has been recognized for a long time and has received increasing attention in recent years. Both local and general noxae can act as trigger and set off a rejection reaction. Epithelial, endothelial and uveal forms of rejection reaction are recognized. The sooner the immune reaction is recognized and the correct therapy started, the greater the change that graft will become clear again.


Glutathione peroxidase: fact and fiction.

The present knowledge of glutathione (GSH) peroxidase is briefly reviewed: GSH peroxidase has a molecular weight of about 85,000, consists of four apparently-identical subunits and contains four g atom of selenium/mol. The enzyme-bound selenium can undergo a substrate-induced redox change and is obviously essential for activity. In accordance with the assumption that a selenol group is reversibly oxidized during catalysis, ping-pong kinetics are observed. Limiting maximum velocities and Michaelis constants, indicating the formation of an enzyme-substrate complex, are not detectable. The enzyme is highly specific for GSH but reacts with many hydroperoxides. It can be deduced from the kinetic analysis of GSH peroxidase that in physiological conditions removal of hydroperoxide is largely independent of fluctuations in the cellular concentration of GSH. However, the system will abruptly collapse if the rate of hydroperoxide formation exceeds that of regeneration of GSH. By these considerations, the pathophysiological manifestation of disorders in GSH metabolism and pentose-phosphate shunt may be explained. With regard to its low specificity for hydroperoxides, GSH peroxidase could be involved in various metabolic events such as H2O2 removal in compartments low in catalase, hydroperoxide-mediated mutagenesis, protection of unsaturated lipids in biomembranes, prostaglandin biosynthesis, and regulation of prostacyclin formation.


Gene clusters and the HLA system.

The major human histocompatibility, or HLA system, is now known to include a number of closely linked loci controlling cell surface specificities, components of the complement system and immune response. Three loci, HLA-A, B and C, control serological determinants found on nearly all cells; a fourth locus, HLA-D, codes for determinants controlling the mixed lymphocyte culture reaction that are present only on B lymphocytes and monocytes. The products of the HLA-A, B and C loci are very similar to each other, as expected if they have arisen by duplication from a common ancestor. The HLA-D products however, show no homology with these and each in turn is different from the complement components (C2, C4 and Bf) coded for in the HLA region. If this complex genetic region has arisen by a series of duplications, one must explain how four different structural gene products can arise from a single duplicated nucleic acid sequence. Different proteins could be translated from the same nucleotide sequence through the use either of different reading frames or of different, not necessarily adjacent, parts of a sequence in different combinations. It seems likely that the general organization of the genetic material of higher organisms is into clusters of duplicated genes like the HLA region and that the numbers of such clusters, rather than the number of individual genes, reflects the genetic complexity of higher organisms.


[Contact eczema caused by 1,4-bis-chloromethylbenzene].

Approximately 3 days of contact to 1,4-bis-chloromethylbenzene of a 24-year old chemistry student led to a bullous contact eczema within approx. 10 days. Patch tests with the substance proved definitely positive. A weak reaction also occurred with 1,4-bis-mercaptomethylbenzene, for the synthesis of which the chloromethyl derivative had been synthesized.


[A prospective multi-centre study of the response of metastatic gastrointestinal tumours (author's transl)].

In a prospective, multi-centre, randomized study of 109 patients with metastatic gastro-intestinal adenocarcinomas the response rate, survival time and side-effects of two drug combinations, carmustin +5-fluorouracil and carmustin + ftorafur, were compared (same carmustin dosage in both groups). Response to the treatment was 32.7% in those receiving carmustin +5-fluorouracil, 26.3% in those on carmustin + ftorafur. This difference occurred among the 42 patients with gastric adenocarcinoma (33.3% compared with 25%), as well as in 11 with pancreatic adenocarcinoma, and in 56 with colorectal adenocarcinoma (32.1% and 28.6%). Median survival time for 5-fluorouracil + carmustin was 330 days, double that for ftorafur + carmustin (163 days). Bone-marrow toxicity (leukopenia, thrombopenia) was below 10% for both drug combinations. Alopecia occurred in only a few patients. Gastro-intestinal toxicity was common (20% and 18.5%, respectively), but there was no difference between the two groups. The somewhat lower effectiveness of ftorafur compared with 5-fluorouracil was probably due to the deliberately smaller dosage of the former.


[The use of organic bone matrix as material for direct pulp capping].

The effect of bone matrix soaked in thyrocalcitonin on vitally amputated pulp was studied in beagle dogs with light and electron microscopy. All treated teeth were vital after eight weeks; the area of implantation was densely populated with cells. Formation of hard tissue differing from normal dentinogenesis had begun.


[Visual tracking disorders in schizophrenia. Review of the literature and methodological considerations].

Since Holzman expressed the hypothesis that eye-tracking dysfunctions might be a genetic indicator of schizophrenia, the researches on the performances of schizophrenics in eye-tacking tasks have been numerous. The correlation between the results of all these researches leaves little doubt on the existence of eye-tacking dysfunctions in schizophrenics. But these distrubances must be precised. Up to now, the studies have used tests that were too complex in their interpretation (pendulum test) and they have been limited to a global appreciation of eye-tacking. To take into account the psychological parameters that may interfere with success in this type of tests, the coming researches should focus on elementary eye-motor activities.


Immunohistochemical visualizations of prolactin, growth hormone, and a substance resembling placental lactogen in the in situ and ectopic pituitary in the hamster.

The effects of transplanting neonatal adenohypophyseal tissue into the hamster cheek pouch on the presence of intracellular materials related to PRL in the allografts were examined using immunohistochemistry. In situ pituitary and placental tissue were used as control. Cells with antirat PRL-reactive sites were scarce in the grafts, and radioimmunoassayable serum PRL was not detectable in hypophysectomized hosts with grafts. Antirat GH- and antihuman placental lactogen (hPL)-reactive sites were visualized in the grafts and in situ pituitary tissue. Intracellular material in hamster placental tissue was visualized with anti-hPL only. Results of various immunohistochemical procedures using in situ pituitary tissue and antirat GH and anti-hPL antisera indicated that three cell types could exist: 1) a cell type visualized with only antirat GH, 2) a cell type visualized with only anti-hPL, and 3) a cell type, the most frequently observed, visualized with both antisera. The physiological significance of this intracellular product, which resembles hPL and perhaps hamster PL immunologically, in hamster adenohypophyseal cells is unknown. Additionally, our data indicate that the ectopic site for pituitary transplantation in the hamster may influence the cell types present in the grafts.


Cholinergic and alpha-adrenergic stimulation of prostaglandin release by dog thyroid in vitro.

Dog thyroid slices released in their incubation medium prostaglandins E2 (PGE2), F2 alpha (PGF2 alpha), 15-keto-13,14-dihydro-F2 alpha, and thromboxane B2 (TxB2), as measured by direct RIA. This release could be inhibited by indomethacin and naproxen, indicating that it corresponded to a neosynthesis. Carbamylcholine (2--100 microns) stimulated the release of PGE2, PGF2 alpha, and TxB2, whereas epinephrine (20 microns to 1 nM) enhanced the release of PGE2 and PGF2 alpha but had no effect on TxB2. These stimulations were inhibited by atropine and dihydroergocryptine, respectively, suggesting that a muscarinic and an alpha-adrenergic receptor were involved. The ionophore A23187 reproduced these stimulations, and the stimulatory effects of carbamylcholine and epinephrine were inhibited in the absence of exogenous Ca++ and after EGTA depletion. Ca++ thus appears as a major regulator of PG production in the thyroid. TSH (0.06--10 MU/ml) and dibutyryl cAMP had no effect on the release of PGE2, PGF2 alpha, or TxB2 by dog thyroid in vitro.


Inhibition by iodide of the cholinergic stimulation of prostaglandin synthesis in dog thyroid.

Iodide is shown to inhibit the cholinergic stimulation of prostaglandin E2 (PGE2) and F2 alpha (PGF2 alpha) synthesis in dog thyroid slices. The inibitory effect of iodide was already detectable at 1 micron and reached its maximal level at 10 microns. At this concentration, iodide inhibited the carbamylcholine-stimulated release of PGE2 and PGF2 alpha by 59% and 73%, respectively (eight experiments). The effect of iodide was neither immediate nor rapidly reversed after a change of incubation medium. The unstimulated release of PGE2 and PGF2 alpha and the stimulation by epinephrine and ionophore A23187 were not modified by iodide in concentrations up to 0.1 mM. The effect of iodide was suppressed in the presence of methimazole (0.1 mM) but not NaClO4 (2 mM). Iodid (0.1 mM) did not inhibit the stimulation by carbamylcholine of PGF2 alpha and PGE2 release by rat pancreas in vitro. These data demonstrate a new action of iodide on the thyroid and establish a link among iodide metabolism, PG synthesis, and cholinergic action in the dog thyroid.


Immunohistochemical interaction on antisera to HCG and its subunits with chorionic tissue of early gestation.

Immunohistochemical localization of hCG and its subunits in chorionic tissue of early gestation was carried out. Antibodies to purified hCG and its subunits were obtained by using these agents for immunization according to the small doses method. The antibody titers and specificities were examined by B/T and standard curves in homologous radioimmunoassay system. The tissue preparations were stained both by a direct and by an indirect method utilizing these antisera and observing the specimens under a fluorescent microscope. The results were as follows. 1) With the anti-hCG staining, fluorescence was observed in the syncytiotrophoblasts as reported previously while the cytotrophoblast were stained slightly. 2) with the anti-hCG-beta staining, the fluorescence was almost identical with that of hCG and showed a more distinct pattern. 3) with the anti-hCG-alpha staining, the fluorescence was found both in the syncytio- and cytotrophoblasts concurrently. Fluorescence of the latter cells was recognized as due to free alpha-subunit because cytotrophoblast was scarcely stained with anti-hCG and anti-hCG-beta.


A genetically diabetic model "KK-CAy mice" for a pharmacological assay.

A genetically diabetic model, KK-CAy mice which were bred by mating female KK mice (aa, BB, cc) with male KK-CAy mice (Aya, BB, CC) was studied on the usefulness as a tool for a pharmacological assay. Body weights of KK-CAy mice increased more rapidly than those of control mice, KK-C. When the body weights of male KK-CAy mice reached about 30 g 10 weeks after birth, their blood glucose levels increased. Severe hyperglycemia (over 300 mg/100 ml) was often observed in the males, but not in the females. Glucose tolerance in the KK-CAy mice was more markedly impaired than that in the control mice. The increase in blood FFA level correlated with the increase in body weight on both KK-CAy mice and the controls. On hyperinsulinemia observed, the ratio of plasma immunoreactive insulin (IRI) level to blood glucose level in the male mice was lower than that seen in the female mice. On hyperglucagonemia observed, elevation of plasma immunoreactive glucagon (IRG) was more remarkable in the males than in the females. Morphological study showed insular degranulation only in the males. Since the dose-dependent insulin-induced falling was observed on blood glucose level in nonfasted KK-CAy mice, they could be used as a feasible tool for an assay of antidiabetic drugs.


Alloxan inhibits glucose oxidation in the pancreatic islets.

The effects of alloxan on glucose oxidation and the protection by anomers of D-glucose from alloxan inhibition of glucose oxidation in the pancreatic islets were investigated using in vitro incubation of rat isolated islets. The pretreatment by alloxan (5-30 mg/dl) for 6 minutes inhibits significantly 14CO2 formation from 14C-U-D-glucose (10 mM) and the addition of alpha-anomer of D-glucose (8.3 mM) to alloxan (20 mg/dl) completely reverses alloxan inhibition of glucose oxidation. These findings seem to be incompatible with the recent view that alloxan acts at the glucose receptor on the plasma membrane of pancreatic beta-cells without affecting glucose metabolism in the pancreatic islets.


Effects of angiotensin I-converting enzyme inhibitor, SQ 14225, in nomal men.

Effects of an orally active angiotensin I-converting enzyme inhibitor, SQ 14225, on the actions of angiotensin I (AI) infused intravenously for 120 to 390 min were studied in 5 normal men. When 20 ng/kg/min of AI infusion was started immediately after a single oral administration of 100 mg of SQ 14225, a significant rise in blood pressure (BP) was observed for the first 15 min, but BP began to fall from 17 min and returned to the pretreatment level at 45 min. This BP level continued at least to 120 min and in one subject to 180 min. In this subject BP began to rise again from 185 min and reached the level of 15 min at 390 min. Plasma AI level increased gradually from 45 min. At 15 min plasma renin activity (PRA) decreased and plasma aldosterone (PA) increased, but then PRA began to increase and PA began to decrease. At 120 min the values of PRA and PA were similar to the pretreatment values. In one subject plasma AI and PRA began to decrease and PA began to increase after 120 or 180 min. On the other hand, in the 5 men sole AI infusion caused a continued BP rise, PRA decrease and PA increase, and sole SQ 14225 administration caused increases in plasma AI and PRA and a decrease in PA but no BP change. From these results it was concluded that complete blockade and partial inhibition of AI conversion by 100 mg of oral SQ 14225 lasted for about 2.5 and 6.5 hr, respectively and that BP rise, PRA suppression and aldosterone stimulation after AI infusion were entirely due to the actions of angiotensin II converted from AI.


Antiepileptic properties of clobazam, a 1-5 benzodiazepine, in man.

Clobazam is a benzodiazepine with special molecular structure (its nitrogen radicals are in positions 1 and 5, rather than 1 and 4 as in all other antiepileptic benzodiazepines), and it is rapidly effective--in a matter of hours or within a few days--against all varieties of epileptic seizures in 52% of subjects treated with it. Its effects are relatively mild. Unfortunately, its outstanding antiepileptic properties are exhausted after only a few weeks in one-third of all cases. The authors discuss the potential significance of this phenomenon, and stress the urgent need for intensive study of the basic mechanism governing exhaustion of the antiepileptic properties of the benzodiazepines in general and clobazam in particular.


Possible modification of scar tissue by biochemical methods.

This paper reviews some of the biochemical modifications involved in fibrous tissue formation and discusses possible ways of controlling fibrosis in clinical conditions. The lathyritic agents, beta-aminoproprionitrile (BAPN) and penicillamine, appear in certain situations to be able to control fibrosis by blocking the biosynthesis of collagen. There are no compounds that are yet known which are capable of reversing pre-existing fibrosis and future research may perhaps be more profitably directed towards the stimulation of collagen catabolism rather than the inhibition of its synthesis.


Fixation of skin grafts in the horse using stainless steel staples.

Three horses with a chronic wound on the distal part of a leg were treated successfully by grafting. Small split skin grafts were fixed onto pieces of adhesive tape. The tape pieces were spread over and fixed to the granulation surface with stainless steel staples. A tight pressure bandage including strongly compressed cellular rubber was then applied over the wound. The combination of staple fixation and strong pressure proved effective in immobilising the skin graft. It was stressed that a firm covering of granulation tissue was a prerequisite for success and therefore the technique should not be used for fresh wounds.


The mechanics of wound healing.

This paper discusses the factors which affect the healing of wounds at the tissue and organ levels. It covers some of the problems which complicate the sequence of healing and considers the mechanisms involved in regeneration and repair of tissues. The factors associated with the stimulus and sequence of healing and their interactions are also reviewed.


Collagen in normal and abnormal tissues.

Despite being a very widespread protein, collagen is an unusual molecule possessing a great tensile strength conferred by a rope-like structure and intermolecular crosslinks. Our current knowledge of the biosynthesis of collagen is providing some insights into certain diseases of connective tissue and is also helping us to understand the healing processes of wounds and diseased tissues.


Outer membrane proteins of smooth and rough strains of Proteus mirabilis.

The outer membranes of the smooth Proteus mirabilis S1959 strain and its rough R13, R110, R51 and R45 mutants were isolated by sonication of the cells and sucrose density gradient centrifugation. The outer membrane of the rough strains had a lower density than that of their parent smooth strain, but the protein-to-phospholipid ratios were the same. The electrophoretic patterns of outer membrane polypeptides of the S and R strains in sodium dodecylsulfate/polyacrylamide gels were identical, with two major polypeptide bands, C1 and C2 (Mr 39,000 and 38,000) predominating. The C1 polypeptide band was a heat-modifiable polypeptide, which migrated as a band at Mr 33,000 when membranes were solubilized at 37 degrees C or 50 degrees C, and at Mr 39,000 when solubilization was at 100 degrees C. Susceptibility of outer membrane polypeptides to proteolytic digestion was found to be higher in isolated outer membrane preparations of the rough strains than in the smooth strain, suggesting that the availability of the polypeptide chains to proteolytic activity depends on the length of the polysaccharide chains of the outer membrane lipopolysaccharide.


External and internal forms of yeast aminopeptidase II.

1. Intact cells of Saccharomyces cerevisiae catalyze the hydrolysis of various aminopeptidase substrates. This activity is not due to permeation of substrates and products but exerted by an external enzyme. 2. From its substrate specificity and the effects of pH and inhibitors the enzyme was identified as aminopeptidase II. 3. About 40% of total aminopeptidase II activity is detectable with untreated exponentially growing cells. Up to two thirds of the external enzyme is released into the medium during enzymic digestion of the cell wall, while little enzyme is liberated by osmotic shock. Membrane preparations contained only small amounts of aminopeptidase II; thus, the localization of the external enzyme appears to be similar to that of the so-called 'periplasmic' yeast hydrolases. 4. By cytochemical methods the presence of aminopeptidase II in the cell envelope was visualized. 5. In contrast to aminopeptidase II, yeast dipeptidase is an entirely intracellular enzyme.


Peptide bond formation stimulated by protein synthesis factor EF-P depends on the aminoacyl moiety of the acceptor.

Elongation factor EF-P is a soluble protein that stimulates peptide bond synthesis catalyzed by the 50-S ribosomal subunit. This factor was previously identified and characterized based on its ability to promote the synthesis of formylmethionine-puromycin. In the present work, we tested the ability of EF-P to promote peptide bond synthesis between ribosome-bound fMet-tRNA and several analogues of the 3' terminus of aminoacyl-tRNA, i.e. the cytidylyl(3'-5')-[2'(3')-O-L-aminoacyladenosines]. EF-P promoted synthesis to the greatest extent with certain acceptors which were otherwise inefficient in the peptidyl transferase reaction. This activity of EF-P could not be replaced by the other soluble proteins known to be involved in polypeptide synthesis, such as EF-Tu, EF-Ts and EF-G. One role of EF-P in protein synthesis may be to allow peptide bond synthesis to occur more efficiently with some aminoacyl-tRNAs that are poor acceptors for the ribosomal peptidyl transferase.


Effect of high-dose prednisone on renal hemodynamics of normal and transplanted dog kidneys.

The effect of prednisone therapy on renal hemodynamics was studied in normal, autotransplanted as well as homotransplanted dog kidneys. After intravenous administration of prednisone (8 and 16 mg/kg/min) the total blood flow decreased and vascular resistance increased significantly. This effect was more pronounced if the drug was directly injected into the arteria renalis. The same changes were observed in autotransplanted and homotransplanted kidneys and in kidneys in rejection. Our results suggest that high dosage of prednisone decreases the renal blood flow and this adverse effect has to be considered in the therapy. The administration of the drug directly into the arteria renalis may have to be avoided.


Hematopoiesis of hereditarily asplenic-athymic (lasat) mice.

The hematopoiesis of athymic-asplenic (lasat) mice was compared with that of normal, asplenic, and athymic littermates with the same strain background. Erythrocyte blood volume, number and survival time were normal when related to the body weight of the animals. Peripheral blood showed leukopenia with absolute and relative lymphopenia, resembling the athymic rather than the asplenic pattern. The bone marrow was hypocellular as a consequence of a decrease in both lymphocytes and erythroid precursors, while thrombocytopoiesis and granulcytopoiesis-monocytopoiesis were essentially normal. Although the percentile value of femoral stem cells was high, their absolute number was, in fact, reduced by 35% as a result of the bone marrow hypocellularity. When lasat bone marrow cells were injected into normal, lethally irradiated mice, a rapid erythropoietic recovery was observed, whereas the restoration of the granlocytic compartment was impaired. It was concluded that: 1) lasat mice depict a normal hematopoiesis in spite of the congenital absence of the thymus and the spleen; 2) bone marrow stem cells may be defective when administered to lethally irradiated hosts; and 3) the athymic status predominates over the asplenic one.


Post-capillary venules of the mouse lymphatic tissues with special reference to the distribution of their endothelial IgG.

Post-capillary venules (PCV) of the lymph nodes and Peyer's patches were studied in frozen sections stained with an indirect immunoperoxidase technique to demonstrate their endothelium-associated IgG in DBA/2 mice rendered T-lymphocyte depleted with anti-theta- and anti-thymus-globulins. Three types of IgG-distribution (luminal, intraendothelial and basement membrane, types I, II and III, respectively), each confined to one of the three PCV grades (graded on the basis of the endothelial cell height), could be distinguished. The PCV grades, on the other hand, were closely correlated with the degree of T-lymphocyte depletion. The significance of this intimate relationship between the IgG-distribution pattern and the PVC grades was discussed in regard to the T-lymphocyte recirculation. A hypothesis was presented describing the presumed sequence of events involved in the passage of T-lymphocytes through the endothelium of the PCV which is the crucial step in their recirculation from blood of the lymphatic tissue.


A morphological study in germfree mice (Salmonella infection).

The morphological manifestation of the intestinal infections of Salmonella typhimurium by the intestine rolled method in both conventional DKI and germfree mice have been investigated in our laboratory. In the conventional mice, the morphological manifestation of intestinal infection shows no difference from the general features of infection, while in the germfree mice, the manifestation is characterized by the so-called non-reactive necrotizing enteritis without cellular reaction. When infection in the germfree mice occurs, some state of immunological insufficiency should be taken into consideration. In infections of germfree mice it is difficult to consider the immunological or the allergic reaction which is expected in the conventional mice. In addition, the direct influences of bacteria such as endotoxin must be taken into consideration as another possibility, but the very limited localized lesions, as seen in this experiment cannot always be suggested to be due to the direct influence of bacteria or intoxication. Though there are many problems to be solved in the future, we are now able to state a kind of mechanism similar to that of Shwartzman's phenomenon cannot be ignored in the infection in newborns or in immunologically immature animals.


The effects of short-term exposure to crowding stress on the pancreatic islets morphology and glycemia in mice.

Behavior, pancreatic islets morphology and plasma glucose levels of male mice exposed for 2, 4, 24 and 48 hours to crowding stress were investigated. The crowding induced an intense turmoil state associated with enhanced irritability and aggressiveness among the specimens of all experimental groups. Violent fights occurred especially in the first 4--6 hours, generally with the death of 1--2 individuals from each group. The changes recorded in the pancreatic islets affected first (2 hours) exclusively the insulin-producing cells, and in subsequent intervals they progressively expanded over all cell types. The changes occurred during the experiment in all islet cell types; however the B-cells showed by far the most pronounced alterations irrespective of the studied time interval. Most changes suggested the stimulation of the entire gland secretory activity, but particularly of B-cells, which was also proved by low glycemia values recorded at 3 of the 4 crowding time intervals. On the other hand, some alterations, occurring first at 24 hours, were regarded as signs of a moderate B-cells secretory hypoactivity; they may partly support the slight hyperglycemia obtained at this time interval. The significance of the above short-term observations in the induction of glycoregulation disturbances, diabetes included, as well as the presumably mediation role of adrenal-cortex and -medulla hormones under stress conditions are discussed and correlated with findings reported in literature.


Fluorophores as visualization aides in agar growth media.

A survey of 26 fluorophores revealed 5 which were non-inhibitory to Staphylococcus aureus and Escherichia coli and produced low background and high colony fluorescence.


Effects of cortisone and thyroxine on intestinal trehalase activity in infant mouse.

Cortisone acetate (25 microgram/g b.wt/day) administration to 8-day-old suckling mice induces a premature increase of trehalase activity along the entire small intestine. On the other hand, thyroxine (1 microgram/g b.wt/day) in unable to provoke a precocious increase of trehalase activity. Trehalase appears to be the only brush border membrane disaccharidase controlled solely by glucocorticoid hormones during the postnatal maturation of the intestine.


Antimicrobial effect of bis-quaternary ammonium salts derived from 1,3-propanediamine.

Antimicrobial activity of bis-quaternary ammonium salts derived from 1,3-propanediamine and 1,3-diamino-2-propanol is described. Effect of the length of alkyl chain and the substitution in the connecting chain on this activity was studied.


An immunological approach to cell separation of the central nervous system.

In this paper I review the various steps of development required for an immunological approach to cell separation. Key points include choice of brain region, isolation procedure, ligand selection, functional assays for separated cell population, and selection of separation procedure. I review briefly some results we have obtained using cerebellar cell populations isolated from 10-day-mice.


Analysis of inherited epilepsy using single locus mutations in mice.

The neurological expression of mutations at defined gene loci in isogenic mice provides a singular opportunity to investigate the developmental pathophysiology of inherited central nervous system (CNS) diseases. Analysis of the single locus mutants that are currently available shows that CNS diseases that include spontaneous seizures as symptoms can be inherited as simple recessive traits. Mutant gene dose is highly correlated with the spontaneous occurrence of seizures. Single gene defects at one of multiple chromosomal loci may give rise to similar epileptic patterns. One mutation, tottering (tg, chromosome 8, recessive) produces in young mice a focal motor seizure pattern with a somatotopic progression, and behavioral absence seizures accompanied by abnormal bursts of bilaterally synchronous, spike-wave discharges in the electrocorticogram. Spontaneous electrographic and clinical seizures of this general pattern bear close resemblance to common forms of human epilepsy. Defined alterations in restricted neuronal pathways of the mouse brain produced by single locus mutations can be used to infer general principles of inherited epileptogenesis, and may provide specific biological test systems for the development of more selective chemical antagonists of seizure activity.


[Changes in carbohydrate digestion enzymes and liver glycogen during heat adaptation].

Adult white rats were exposed daily during 30 days to 2-hour heating (40--41 degrees)--adaptation, and then were transferred to ordinary temperature conditions at 25--27 degrees--desadaptation. The initial period of adaptation revealed a decrease of the liver glycogen content and a relatively constant level of glycemia. A considerable activation of the carbohydrate digestion enzyme systems followed this lowering of energy resources. Metabolic regulatory mechanisms seem to take part in this activation which plays an adaptive role in maintenance of constant glycemia level under these stressful conditions.


The phagocytosis of Candida albicans blastospores and germ tubes by polymorphonuclear leukocytes.

This in vitro study investigates the phagocytosis and killing rate of Candida albicans blastospores and germ tubes by polymorphonuclear leukocytes. Suspensions of C. albicans blastospores and germ tubes were incubated for 60 and 150 min with a neutrophil suspension. The rate of phagocytosis was found to be 92 +/- 3% for blastospores, but only 9.5 +/- 2.5% for germ tubes. The candidacidal activity rate was 29 +/- 6% for blastospores, but only 5.5 +/- 1.5% for germ tubes. Electronmicroscopically, cytoplasmic and plasma membrane alterations of phagocytized yeast cells lying in phagosomes were observed. Short germ tubes surrounded by a phagosomal membrane were found in neutrophils. Older, longer germ tubes were seen in an extracellular position. Usually, several neutrophils were adjacent to these long tubes. Obviously phagocytosis could not take place owing to the size of the germ tubes. The findings indicate that the transition of the yeast phase to the mycelium phase in a case of Candida infection may be a mechanism enabling the parasite to escape phagocytosis by the host.


Circulating antibodies in malignant melanoma patients.

Sera from 46 patients with malignant melanoma (MM) in different clinical stages and from 25 healthy blood donors were studied by an indirect conventional immunofluorescent technique (IIT) and by an immunofluorescent complement-fixation test (ICFT). No specific and reliable serological reactivity could be demonstrated against autologous and homologous MM and other nonmalignant pigmented tissues. An increased incidence of diffused antinuclear (ANA; 41%) and cytoplasmic epidermal cell autoantibodies (A-CYT; 71%) was observed in the patients by IIT. Statistical evaluation of the results gave significant values in the chi 2 test in patients for ANA (chi 2 = 5.2061; p less than 0.05) and for A-CYT (chi 2 = 8.9176; p less than 0.01) versus age-matched controls. Thyroid microsomal, gastric parietal cell, smooth muscle and mitochondria autoantibodies were, respectively, detected in 4.3, 4.3, 4.3 and 0% of patients' sera.


Lichenoid dermatitis due to chlorpromazine phototoxicity.

A 75-year-old woman had lichenoid and bullous lesions in the sun-exposed areas. She had a history of cerebral embolism and received chlorpromazine (CPZ) until consultation at our clinic. Patch tests with CPZ were negative, but photopatch tests were positive at relatively high concentrations of CPZ and ultraviolet light radiation sites of 310, 340 and 370 nm. Rapid improvement of the lesions occurred upon discontinuation of CPZ and the avoidance of exposure to sunlight. From these results, the patient was considered to show lichenoid eruptions due to CPZ phototoxicity rather than photoallergy.


[Manifestation of a malignant lymphoma of high grade malignancy in the terminal stage of mycosis fungoides. Enzyme-cytochemical and immunocytological investigations (author's transl)].

Report on a patient with mycosis fungoides, in whom after a long course the disease developed rapidly into a malignant lymphoma of high grade malignancy. The diagnosis was confirmed by cytomorphological and enzyme-cytochemical methods (demonstration of hydrolytic enzymes in cryostat sections, cutaneous smears and cell suspensions extracted from skin lesions), as well as by immunocytological methods (differentiation of lymphocyte sub-populations). In the tumour stage of mycosis fungoides, a polymorphous infiltrate composed of lymphocytoid and monocytoid cells prevailed, while a proliferation of lymphoblastic cells was present in the terminal stage of the disease. These cells showed no longer the properties of mature lymphocytes and differed also in their enzyme-cytochemical pattern.


Immunological and histopathological studies in a case of pyodermia vegetans Hallopeau.

In a case of pyodermia (pemphigus) vegetans Hallopeau, modern investigations were performed. Direct immunofluorescent (IF) study of specimens taken from verrucous vegetations failed to reveal the presence of IgG deposits in the intercellular spaces as well IgA and IgG deposits at the epidermal-dermal junction. Indirect IF examination of circulating pemphigus antibodies gave a negative result. Histopathological study did not show the presence of acantholysis. The studies which were carried out indicated that pemphigus vegetans Hallopeau should not be associated with the pemphigus group.


Immunohistochemistry in lichen planus.

An immunohistochemical study has been undertaken on 25 biopsy specmens taken from lichen planus lesions, using antisera against human fibrin, immunoglobulins IgG, IgA, IgM and C3 (B1C/B1A) complement component. The findings of the present research are discussed and evaluated in relation to the problem of the etiopathogenesis of the disease.


Comparison of in vivo and in vitro capability of complement fixation by pemphigus antibodies.

Using complement immunofluorescence, the capability of complement fixation by bound antibodies in pemphigus lesional skin was compared with circulating intercellular antibodies of complement-fixing type in the same patient. Five lesional skin samples from 6 patients, containing bound IgG, gave positive binding of complement (C3) supplied from fresh guinea pig serum, while only one serum sample showed complement-fixing pemphigus antibodies in the circulation. These results support the view that complement-fixing pemphigus antibodies are rapidly localized to the skin and play a role on the production of acantholysis.


Controlled clinical trial of a rifampicin- trimethoprim combination (Rifaprim) and cotrimoxazole in patients with chronic urinary tract infections.

A controlled clinical trial of Rifaprim (rifampicin 600 mg plus trimethoprim 160 mg, in a single daily dose at 10 p.m.) and cotrimoxazole (two tablets b.i.d.) was carried out in two groups of twenty-one patients, each with chronic or recurrent urinary tract infections. Both treatments lasted 10 days. At the first follow-up, 3-11 days after the end of treatment, bacteriological failure was observed in two Rifaprim patients and in four contrimoxazole patients. Clinical improvement with sterile urine or markedly reduced bacterial count was observed in nineteen. Rifaprim patients and in fifteen cases of the other group. Mild allergic phenomena were observed in three Rifaprim patients, but they did not require discontinuation of treatment; anorexia was complained of by a cotrimoxazole patient. Rifaprim appears a valuable alternative in the treatment of chronic urinary tract infection, mainly because it reduces or abolishes the risk of emergence of resistance.


Effect of norepinephrine on insulin, glucagon, and somatostatin secretion in isolated perifused rat islets.

The rate of insulin, glucagon, and somatostatin secretion was measured from isolated rat islets maintained in a perifusion system. The effect of norepinephrine (NE) was simultaneously determined on the release rate of all three hormones. Norepinephrine was employed at an acute dose of 10 micrometers and in graded doses from 1 nM to 10 micrometers in the presence of high (22 mM) and low (1.4 mM) glucose conditions, insulin secretion was maximally inhibited at 10 micrometers NE concentration and was significantly depressed at 100 mM NE concentration. Under both high and low glucose conditions, glucagon release was maximally stimulated at 10 micrometers NE concentration and was significantly elevated at 10 nM NE concentration. Under high and low glucose conditions, somatostatin release was inhibited by 10 micrometers NE concentration and was significantly depressed at 100 nM NE concentration. During the initial maximal stimulation of glucagon, NE inhibition of somatostatin and insulin was prevented, possibly by the high level of glucagon released. A paracrine effect of glucagon on beta and delta cells is proposed.


A study of glucagonomas by light and electron microscopy and immunofluorescence.

Five tumors associated with the complete glucagonoma syndrome, as well as a series of glucagon-cell adenomas from three patients without this syndrome, were investigated by light and electron microscopy and by immunofluorescence. All tumors associated with the syndrome were large, from 3 to 35 cm along the major axis, and three of them were proved to be malignant. No common histologic arrangement of tumor cells was apparent for the five neoplasms examined. Immunofluorescent staining for glucagon and glicentin was carried out: while most cells were negative, a varying number of scattered cells were positive with both antisera in all tumors except one; three tumors contained more glicentin- than glucagon-immunoreactive cells. Moreover, three tumors were multihormonal, witn cells positive for pancreatic polypeptide and/or insulin. Ultrastructurally, the secretory granules of cells from these tumors were not typical of those found in A-cells from adult human islets. The glucagon-cell tumors from patients without the syndrome were benign, usually multiple, and were small, with diameters from 0.5 mm to 1 cm. In most cases, the cells from these neoplasms arranged in a characteristic pattern (ribbonlike or "gyriform"). In most tumors, the majority of cells showed both glucagon and glicentin immunofluorescence and the ultrastructural appearance of their secretory granules was similar to that of normal islet A-cells. From the morphologic point of view, therefore, cells from tumors not associated with the glucagonoma syndrome resemble normal glucagon cells more closely than those from tumors associated with the syndrome.


Immunofluorescence study of mucosal B lymphocytes in bile reflux gastritis.

Intracelluar immunofluorescence staining was used to identify lymphoid cells from human gastric mucosa. Staining of serial sections for each of the five classes of immunoglobulin-containing cells revealed the presence of all classes. Their relative frequencies in the antral mucosae of 30 patients with bile reflux gastritis were 51% IgA, 16% IgM, 18.5% IgG, 2.5% IgD and 12% IgE. The mucosae of 15 control subjects showed 75% IgA, 8% IgM, 15% IgG, 1% IgD and 1% IgE and the mucosae of 30 patients with inflammatory gastritis without bile reflux revealed 67% IgA, 6.5% IgM, 22.5% IgG, 1.5% IgD and 2.5% IgE. These features suggest that allergy participates in the physiopathology of bile reflux gastritis.


Modulation of human gastric mucosal adenylate cyclase activity by prostacyclin.

Prostacyclin, its stable metabolite 6-keto-prostaglandin F1 alpha and prostaglandin E2 were tested on the adenylate cyclase system in human gastric mucosa. All three compounds were able to activate human cyclase. Saturating concentrations of prostacyclin, prostaglandin E2 and 6-keto-prostaglandin F1 alpha (each 0.28 mM) increased the enzyme activity 3.0 and 1.6 fold, respectively. The concentrations of prostacyclin and prostaglandin E2 required to produce half maximal activation were in the same range. Prostacyclin is an efficient stimulator of human gastric mucosal adenylate cyclase.


Simultaneous expression of two different types of intermediate sized filaments in mouse keratinocytes proliferating in vitro.

The intermediate-sized filaments present in epidermal keratinocytes derived from mouse skin and in an established cell line (HEL) derived from spontaneous transformation of murine keratinocytes grown in vitro, have been examined by immunofluorescence microscopy, using antibodies directed against subunit proteins of different classes of intermediate-sized filaments, as well as by electron microscopy and gel electrophoresis of cytoskeletal preparations highly enriched in intermediate-sized filaments. The keratinocytes derived from neonatal skin, which are capable of only limited replication in vitro, show only a single type of intermediate-sized filaments, i.e., the tonofibril-like arrays of filaments containing prekeratin. HEL cells, which proliferate indefinitely in vitro, retain the tonofilament-like structures typical of differentiated epidermal cells but in addition display intermediate-sized filaments of the vimentin type, i.e., the filament system typically found in mesenchymal and mesenchyme-derived cells. We discuss the possibility that (i) the advent of vimentin-type filaments in epidermal cells in culture is related either to the transformed state or the in vitro growth conditions as such and (ii) other differentiated epithelial cells proliferating in vitro may have more than one system of intermediate-sized filaments.


[Electrophoresis mobility inhibition test (emt) in immunological early diagnosis of gynaecological malignomas (author's transl)].

The electrophoresis mobility inhibition test (EMT) was employed in the examination of a total of 74 woman patients with various gynaecological malignomas, 50 women with non-malignant diseases and 41 healthy female controls. In the group of patients with  malignoma, 78.4% of the test results agreed with the histological diagnosis, 21.6% of the results were false negative. In the group of patients with benign diseases, 78.0% of the test results were correct, whereas 22.0% were false-positive. One false-positive test result was seen among the healthy controls. Extensive tumour growth, chronic infections and virus diseases seem to falsify the test result. Significance, perspectives and limitations of this test system within the framework of tumour diagnosis, are discussed.


[The relevance of "family-oriented obstetrics" and its consequences for the maternity hospital (author's transl)].

By inventing the "rooming in", the organisational prerequisites for the undisyurbed early mother--child relationship, which is of fateful relevance for the newborn, are provided. Since part of the responsibility for the child is transferred to the mother already in the hospital, an adequate instruction is necessary. Thus, a change is taking place pertaining to the tasks of the nursing personnel. In an inquiry made on 950 female patients, the social-psychological aspects of the "romming in" were examined and evaluated. The joint accommodation of mother and child ought to be available on request for approximately 50% of the delivered women. During the night the newborns should be taken to the nursery. This meets with the needs of many women for undisturbed rest during the night despite "rooming in" and also with the necessity for control of the newborns by the nursing staff. The general judgment of the patients on a clinic cannot be changed essentially by the invention of "rooming in". This is depending to a great extent on various other factors.


[Effect of radiosensitivity gene irradiation and genotype on yeast cytoduction].

Effects of gamma- and UV-irradiation of one parent on progenies of yeast crosses are studied. In the crosses of the type: p+ a alc4 X p-alpha ade2 (alc4--nuclear respiration deficiency) zygotic colonies (white) and haploid p+ cytoductant colonies (red) were scored. The irradiation of p+ parent increases the cytoductant frequency up to 17%. When both parents are radiation-sensitive (xrs1, allelic to rad54), the frequency of cytoduction reaches 90%. Crosses p+xrs1 X p-xrs+ or p+xrs+ X p-xrs1 are similar to crosses of wild types. The same results were observed in the case of xrs4 mutants as well as for UV-irradiation. In the progenies of crosses irradiated p+ a alc4 (xrs+ or xrs1) X non-irradiated p-alpha ade2 ura (xrs+ or xrs1) the genotype of red colonies was analysed: in most cases (82.3%-99.8%) they were p+ alpha ade2 ura, that was true haploid cytoductants. It is concluded that irradiation induces particular damages of yeast nuclei leading to a block of normal karyogamy and thus to a cytoductant formation. The highest frequency of cytoduction was observed in crosses of radiosensitive mutants.


[Genetic control of the formation of plasmid F'. I. Effect of recA- and seg-2 mutations in an Hfr donor strain on the character of plasmid F' formation].

Assuming the similarity of the processes of illegitimate recombination, such as deletion formation, with the process of F' plasmid formation, we have undertaken the study of the influence of recA- and seg- alleles of Hfr donor on the F' plasmid formation. The data obtained demonstrate the strong influence of donor genotype on the frequency of F' plasmid formation and on the nature of F' plasmids formed, thus demonstrating that the most of F' plasmids have been formed via recombination in Hfr donor cells. The recA- mutation decreased the total yield of F' plasmids selected using both proximal and distal Hfr markers and affected drastically the distribution of the F' plasmids inheriting different proximal unselected markers. The existence of recA-dependent and recA-independent modes of F' plasmid formation was demonstrated. The Escherichia coli chromosome contains regions which involve preferentially in recA-dependent (between proA and gal, and clockwise from gal) or recA-independent (between leu and proA, and the region counterclockwise from argE) recombination. The seg-2 mutation causes only partial block of both recA-dependent and recA-independent recombination pathways, however it causes dramatic decrease of genetic exchanges leading to the formation of the type II F' plasmids. Both seg- and recA- mutations decrease the frequency of the formation of Tra+ F' transconjugants. The percent of Tra- transconjugants, which remain sensitive to MS2 and Q beta donor specific phages, also drops significantly under the influence of the recA- and seg- alleles. Thus, the recombination involving the F structure in wild type strains and seg- mutants occures preferentially in the points of F outside the regions essential for transfer and sensitivity to male specific phages, while in recA- and recA-ges- strains the points inside these regions (tra operon) frequently involved in F' plasmid looping out. There exist more strict correlation between the fertility and sensitivity to phage Q beta than to phage MS2.


[Enhanced UV sensitivity of Escherichia coli strain uvrA crp].

UV-sensitivity and UV-induced mutability to tryptophan independence has been studied in isogenic crp, cya, crp+, uvrA crp and uvrA crp+ strains of Escherichia coli. crp and cya strains are found to have the same UV-sensitivity as an isogenic wild type strain. UV-sensitivity of uvrA crp strain seems to be one-two orders increased as compared with the sensitivity exhibited by the uvrA - crp+ strain. The yield of UV-induced revertants is slightly higher in crp, cya and uvrA crp strains than in the wild type cells. The existence of cap-dependent inducible error-free repair pathway is supposed due to the data obtained.


Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells.

Escherichia coli cells are 4--6 times more transformable and 20--30 times more competent after 24 h incubation in cold calcium chloride than immediately after calcium chloride treatment. With 24-h-old competent cells we obtained routinely 2 . 10(7) transformants per microgram of pBR322 DNA, and transformed over 20% of viable cells.


Cell-free synthesis of proteins coding for mobilisation functions of ColE1 and transposition functions of Tn3.

A hybrid plasmid, pJC74, carrying the large (3.35 Md) EcoRI-PstI fragment of ColE1, the 1.3 and 1.8 Md PstI fragments of plasmid R1 drd19 (containing a part of Tn3 and known to specify transpostion functions), and a part of gamma bacteriophage DNA carrying the fused cohesive ends of gamma (cos) was constructed. Supercoiled DNA of pJC74 and a series of deletions (series pJC75) were used in a cell-free coupled transcription-translation system. Analysis of the proteins produced allowed the identification of a 28 to 30 kd protein and/or a 3 kd protein responsible for mobilisation of the plasmids in sex-factor-promoted conjugation. Furthermore, three protein bands of 12, 12.5 and 13 kd were correlated with the presence of a portion of the Tn3 transposon previously shown to code for transposition functions which can complement in trans. These latter and the 30 kd beta-lactamase were the only proteins identified as Tn3-specific by comparison of ColE1 with RSF2124.


Restriction map, partial cloning and localization of 9S and 12S kinetoplast RNA genes on the maxicircle component of the kinetoplast DNA of Leishmania tarentolae.

We have constructed a restriction map of the maxicircle component of the kinetoplast DNA of Leishmania tarentolae for the enzymes EcoRI, Bam HI, HaeIII, HpaII, SalI, BglII and HindIII. The 9 and 12S kinetoplast RNAs were localized on this map. Two fragments of this maxicircle molecule were cloned in the bacterial plasmid, pBR322, including a 4.4 . 10(6) dalton EcoRI/BamHI fragment which contains the 9 and 12S RNA genes.


Plasmids with temperature-dependent copy number for amplification of cloned genes and their products.

Miniplasmids (pKN402 and pKN410) were isolated from runaway-replication mutants of plasmid R1. At 30 degrees C these miniplasmids are present in 20--50 copies per cell of Escherichia coli, whereas at temperatures above 35 degrees C the plasmids replicate without copy number control during 2--3 h. At the end of this period plasmid DNA amounts to about 75% of the total DNA. During the gene amplification, growth and protein synthesis continue at normal rate leading to a drastic amplification of plasmid gene products. Plasmids pKN402 (4.6 Md) and pKN410 (10 Md) have single restriction sites for restriction endonucleases EcoRI and HindIII; in addition plamid pKN410 has a single BamHI site and carries ampicillin resistance. The plasmids can therefore be used as cloning vectors. Several genes were cloned into these vectors using the EcoRI sites; chromosomal as well as plasmid-coded beta-lactamase was found to be amplified up to 400-fold after thermal induction of the runaway replication. Vectors of this temperature-dependent class will be useful in the production of large quantities of genes and gene products. These plasmids have lost their mobilization capacity. Runaway replication is lethal to the host bacteria in rich media. These two properties contribute to the safe use of the plasmids as cloning vehicles.


Expression of the yeast galactokinase gene in Escherichia coli.

In Saccharomyces cerevisiae the genes for three of the enzymes involved in galactose metabolism are tightly linked near the centromere of chromosome II (Douglas and Hawthorne, 1964). However, the molecular mechanisms which control the expression of these genes are not well understood. A DNA fragment containing at least one of these yeast genes, the galactokinase gene (gal1), has been joined to the bacterial plasmid pBR322 and maintained in an Escherichia coli strain that carries a deletion in its own galactokinase gene, galK. The presence of the yeast gene was demonstrated by (i) complementation of the E. coli galactokinase deletion, (ii) by hybridization of the cloned DNA fragment to restriction enzyme digests of total yeast DNA and (iii) by assaying for yeast galactokinase activity in bacterial cell extracts. The yeast DNA fragment is 4700 base pairs long, and enables the host E. coli K-12 strain to grow in minimal medium containing galactose as the sole carbon source with a generation time of 14.3 h. The yeast galactokinase activity in the bacterial extracts is 0.7% of the bacterial galactokinase activity found in wild-type E. coli fully induced with fucose.


Plasma renin, renin substrate and angiotensin II changes following experimental endotoxinaemia.

The effect of a single dose of endotoxin (B. coli, 1.5 mg/kg intravenously) on plasma renin concentration (PRC), renin substrate (PRSC), and angiotensin II (AT II) was studied in rats over a period of 48 hours. All determinations were performed by specific radioimmunoassay. Six and nine hours following endotoxin administration, renin secretion was decreased, whereas at 48 hours a slight increase in the PRC was found. In contrast, a three-fold elevation of the PRSC occurred during the first 24 hour period, attributable to a stimulation of the hepatic biosynthesis as result of corticosterone oversecretion. According to the observed changes in PRC and PRSC, AT II remains unchanged after six and nine hours, whereas a significant increase was detected after 24 and 48 hours. Based on the actual AT II level, the findings emphasize that in the rat the RAS does participate in the later stages of endotoxin stress only.


Effects of pinealectomy and pineal incubation medium and sonicates on insulin release by isolated pancreatic islets in vitro.

The present studies were designed to investigate the mechanism of previously-reported nocturnal hyperinsulinemia in the pinealectomized rat. Isolated islets were obtained from anesthetized control, sham-pinealectomized and pinealectomized rats, with 5 rats per surgical groups, during the early dark phase of the daily lightdark cycle. Batches of 3 islets each were incubated in various combinations of 2, 10 or 30 mM glucose with control buffer, medium in which cerebral cortex or pineal glands had previously been incubated for 2 hours, or sonicates of these same tissues. Insulin released into the culture medium was measured by radioimmunoassay. A significant hypersecretion of insulin was demonstrable in the islets from the pinealectomized animals. A stimulatory effect of both pineal medium and sonicates upon insulin release was similarly observed. Neither of these effects displayed an interaction with the concentration of glucose in the islet incubation medium and they, therefore, appear to be mediated by a mechanism which operates independently of stimulation by glucose. These results indicate that the rat pineal gland can exert direct effects upon insulin release from the islets, possibly through a humoral route. Further studies are in progress to characterize the nature and mode of action of the insulinotropic agent present in and released from the pineal gland.


Secretion of glucagon and insulin in hypophysectomized rats: effect of tolbutamide.

Normal and hypophysectomized (hypox) rats, fed ad libitum, received intraperitoneal injections of tolbutamide (75 mg/kg/day) or of saline for 6 weeks. 24 h after the last injection, blood samples were taken for glucose, insulin and glucagon determinations. In normal rats, tolbutamide treatment did not alter serum glucose, insulin and glucagon, although it suppressed the secretion of insulin and glucagon by the pancreatic islets. In hypox rats, tolbutamide decreased serum glucose and insulin, elevated serum glucagon and stimulated the secretion of glucagon, but not that of insulin by the pancreatic islets. In addition, tolbutamide treatment increased the glucagon response to arginine in normal, but not in hypox rats. The serum glucose response to arginine was decreased by tolbutamide treatment and by hypophysectomy and, thus, appeared independent of the glucagon rise or preexisting glucagon level. We conclude that tolbutamide treatment decreased the secretion of glucagon and insulin in normal rats and stimulated that of glucagon in hypox rats, perhaps because of the low levels of insulin in the serum and in the pancreas of the latter. Our results are compatible with the hypothesis that the pancreatic action of tolbutamide is influenced by the pituitary.


Planning process a powerful tool.

Although business and industry have been involved with the planning process for some time, hospitals have only recently realized the powerful tool that the process represents.


Identification of mouse mammary adipose cells by membrane antigens.

Antisera produced to mammary adipose cells from midpregnant BALC/c females can be used to distinguish mammary adipose cells from mammary epithelial cells and fibroblasts. The mammary adipose membrane antigen detected by indirect immunofluorescence was found in adipose cells from (a) mammary glands of virgin, midpregnant and lactating mice; (b) mammary fat pads that had been surgically cleared of glandular elements; and (c) epididymis. In all tissues, this cell-surface antigen was removed by the enzymatic action used to dissociate the cells from the tissues and was shown to be fully restored when cells were cultured for 48 hr.


[Serodiagnosis of malaria. Plasmodium berghei and P. falciparum as antigen for the indirect immune of fluorescence test (author's transl)].

Serum samples of malaria patients were investigated by the IIFT with P. berghei antigen (from mice) and P. falciparum antigen (from in vitro cultures). Both antigens were useful principally and produced approximately identical results. P. falciparum antigen produced mostly higher titres than P. berghei antigen. In cases of lack of homologous malaria antigen it can be suitable to use an antibody-free P. berghei antigen for the IIFT, especially in cases of epidemiological studies. For the individual clinical diagnosis titres of 1:64 are significant.


The antinflammatory effects of glucocorticosteroids. A brief review of the literature.

The effects of glucocorticosteroids on immune and inflammatory responses are reviewed. The steroids seem to change the number or function of cell receptors for regulating agents, so that in areas of inflammation: (a) blood vessels dilate less, (b) lymphocytes proliferate less, (c) all leukocytes infiltrate less, (d) macrophages become less activated (digesting and secreting less), and (e) fibroblasts produce less collagen and ground substance. In addition, the corticosteroids seem to alter the response of cells to various signals from their receptors by affecting the prostaglandin system, cyclic nucleotides, and perhaps other internal mediators.


Specific absorption of human serum albumin, immunoglobulin A, and immunoglobulin G with selected strains of group A and G streptococci.

Five gram-positive bacterial strains were selected for absorption studies of human serum samples. Strain AR1 (group A, M-type 1) and G148 (group G), with strong immunoglobulin G (IgG) binding capacities, and strain AW43 (group A, M-type 60), binding both IgA1 and IgA2, were compared with Staphylococcus aureus Cowan I and with Staphylococcus epidermidis L603. Both AR1 and G148 were capable of completely absorbing out serum IgG. In contrast, S. aureus Cowan I left a fraction unabsorbed, as expected from its known lack of IgG3 binding. Strain AW43 absorbed out all serum IgA, using a 10-microliter bacterial pellet for 20 microliter of serum. Serum IgM levels were slightly reduced by S. aureus Cowan I absorption. On the basis of the experiments, a bacterial mixture was designed consisting of S. aureus Cowan I and group A streptococcus strains AR1 and AW43, with absorption characteristics suitable for use in discriminating between early IgM and late IgG and IgA immune responses in routine serological work. A new type of bacteria-mammalian protein binding was discovered. Human serum albumin was completely absorbed out by strain G148 and to a lesser extent by strain AR1 and AW43. S. aureus Cowan I and S. epidermidis were negative. The binding capacity of G148 for albumin equalled that of Cowan I for IgG. The binding pattern of albumin to the strains was different from those of IgG, IgA, IgM, fibrinogen, haptoglobin, or aggregated beta 2-microglobulin and therefore seems to represent another type of bacterial-mammalian interaction with a specific albumin receptor on the surface of streptococci.


Effects of age, ambient temperature, and heat-stable Escherichia coli enterotoxin on intestinal transit in infant mice.

Some interrelationships among age, ambient temperature, intestinal transit, and enterotoxigenic Escherichia coli infection were studied in an infant mouse model. The transit of dye in the small intestine was accelerated during the response to heat-stable E. coli enterotoxin. Transit in the small intestine of normal mice accelerated with increased age (from less than 17 h to 8 days old) and accelerated with increased ambient temperature (from 25 to 37 degrees C). Transit was more rapid in the jejunum than in the ileum throughout the range of experimental conditions studied. E. coli strains that do not produce any of the pili known facilitate intestinal colonization were cleared from the small intestine more rapidly at 37 degrees C than at 25 degrees C. This clearance was thought to be due to accelerated transit at the higher temperature. In contrast, a strain of E. coli that produces K99 (pili previously shown to facilitate intestinal colonization in other species) was not cleared from the small intestine and colonized more intensively at 37 degrees C than at 25 degrees C. Intensified colonization by this strain was thought to be due to increased production of K99 at the higher temperature. It was suggested that sluggish intestinal transit may also be characteristic of the neonates of other species and be one of the factors predisposing them to intestinal colonization by enteropathogens. It was speculated that this predisposition may be enhanced if the neonates are chilled. However, the effect of ambient temperature on intestinal transit in homeothermic neonates such as pigs, calves, and humans may be different from that in mice because neonatal mice are poikilothermic.


Comparison of assay of coliform enterotoxins by conventional techniques versus in vivo intestinal perfusion.

Thirty-six strains of coliform bacteria were tested for enterotoxigenicity both by conventional assays, including the Y-1 adrenal and Chinese hamster ovary cell assays for heat-labile toxin and the suckling mouse assay for heat-stable toxin, and by determining the ability of graded concentrations of ultrafiltrate high- or low-molecular-weight toxin preparations to induce water secretion during in vivo perfusion in the rat jejunum. The ultrafiltrates of all 18 strains isolated from persons with infectious diarrheal disease, including seven of Escherichia coli, seven of Klebsiella pneumoniae, and four of Enterobacter cloacae, contained one (nine strains) or two (nine strains) potent toxin fractions (resembling either heat-labile or heat-stable toxin in terms of apparent molecular weight and heat lability characteristics) that induced water secretion at perfusion concentrations of 10 ng/ml or less. Unconcentrated broth filtrates of five of the E. coli strains and two of Klebsiella reacted positively in one or more of the conventional assay systems. Concentrated ultrafiltrates from two strains that were negative in the in vitro assays for heat-labile toxin were tested and also proved to be inactive in these test systems. None of 18 strains isolated from control sources produced, in the ultrafiltrates, enterotoxins capable of inducing water secretion at low concentrations, and none reacted positively in the conventional assays. These results indicate that some strains of coliform bacteria elaborate potent toxin materials that are capable of inducing water secretion and can be detected by perfusion of concentrated ultrafiltrates but not by conventional assay systems for enterotoxigenicity. Whether this represents quantitative or qualitative differences between the toxin materials that stimulate these different test systems remains to be established.


Pyruvate oxidation by Treponema pallidum.

Cell-free extracts of Treponema pallidum catalyzed the decarboxylation of pyruvate. This activity was suppressed at low O2 tensions and appeared to be coenzyme A independent. Pyruvate decarboxylation was inorganic phosphate dependent, and evidence suggested that acetyl phosphate was a product. Oxygen was consumed, and data indicated that H2O2 was produced. These results indicated that the overall oxidation of pyruvate was: pyruvate + O2 + inorganic phosphate leads to CO2 + acetyl phosphate + H2O2. Phosphotransacetylase and acetate kinase activities were also observed in the cell-free extracts and could catalyze formation of acetyl coenzyme A and adenosine 5'-triphosphate, respectively, from acetyl phosphate.


Delayed hypersensitivity responses in mice and guinea pigs to Mycobacterium leprae, Mycobacterium vaccae, and Mycobacterium nonchromogenicum cytoplasmic proteins.

Antigenic relationships between Mycobacterium vaccae, M. nonchromogenicum, and M. leprae were examined in mice and guinea pigs injected with M. vaccae or M. nonchromogenicum suspensions. The growth of both organisms in outbred ICR and four inbred mouse strains was followed up to 30 days. M. nonchromogenicum persisted in the livers and spleens of the inbred mice substantially better than did the M. vaccae population in the same mouse strains. A translucent colony variant of M. vaccae isolated from the opossum survived in vivo better than the opaque colony isolated from opossums and cattle. Persistence of M. vaccae and M. nonchromogenicum was not markedly increased in T-cell-depleted (nude) mice. Normal mice infected with increasing numbers of M. vaccae did not develop delayed-type hypersensitivity to the homologous M. vaccae cytoplasmic protein antigen. When heat-killed M. vaccae were incorporated into Freund adjuvant, both mice and guinea pigs developed delayed hypersensitivity to cytoplasmic antigens prepared from M. vaccae, M. nonchromogenicum and M. vaccae vaccines cross-sensitized guinea pigs to the M. leprae cytoplasmic antigens.


Opsonization of Cryptococcus neoformans by human immunoglobulin G: role of immunoglobulin G in phagocytosis by macrophages.

The role of immunoglobulin G (IgG) as an opsonin in phagocytosis of Cryptococcus neoformans by macrophages was investigated. Labeling with 125I showed that IgG isolated from normal human serum bound to non-encapsulated C. neoformans. Furthermore, IgG-opsonized cryptococci were agglutinated by anti-serum to IgG heavy chains, indicating that normal human serum contains antibody that will bind to the yeast surface. The IgG isolated from normal serum accounted for all opsonizing activity found in normal human serum, since differences were not noted between the opsonizing activities of whole serum, heat-inactivated serum and purified IgG when these opsonins were compared at equivalent concentrations of IgG. Phagocytosis of IgG-opsonized cryptococci was inhibited by anti-macrophage IgG, a reagent known to block Fc-mediated attachment and ingestion, and by pepsin digestion of opsonizing IgG. Thus, IgG opsonization is an Fc-dependent process. Opsonizing IgG appears to play its major role during the attachment phase of phagocytosis, since antimacrophage IgG blocked attachment of cryptococci to macrophages but could not block ingestion of IgG-opsonized cryptococci that had been allowed to attach to macrophages. Ingestion of opsonized cryptococci was not blocked by 2-deoxy-D-glucose, a reagent known to block Fc-mediated ingestion, thus confirming that IgG has a primary role in attachment and suggesting that ingestion is mediated by a process that is not Fc dependent.


Opsonization of Cryptococcus neoformans by human immunoglobulin G: masking of immunoglobulin G by cryptococcal polysaccharide.

Previous studies have shown that attachment of non-encapsulated cryptococci to macrophages is highly dependent on opsonizing immunoglobulin G (IgG) and that cryptococcal polysaccharide inhibits the attachment phase of phagocytosis. We investigated various mechanisms by which cryptococcal polysaccharide might interfere with the opsonizing action of IgG. Cryptococcal polysaccharide did not appreciably prevent binding of opsonizing IgG to the yeast. Furthermore, cryptococcal polysaccharide acted as a noncompetitive inhibitor with respect to the opsonizing action of IgG. These experiments suggested that cell wall-bound IgG is masked in some manner such that it is unable to participate in Fc-mediated phagocytosis. This appeared to be the case, since cryptococcal polysaccharode inhibited agglutination of IgG-opsonized yeast cells by antiserum to IgG. There was good dose-response correlation between the amount of polysaccharide needed to inhibit phagocytosis of non-encapsulated Cryptococcus neoformans and the amount of polysaccharide needed to prevent agglutination of IgG-opsonized cryptococci by antiserum to IgG. The ability of cryptococcal polysaccharide to prevent agglutination of IgG-opsonized cryptococci by antiserum to IgG was lost if dextran, a substance known to enhance agglutination of several particles, was incorporated into the medium.


Cryptococcal culture filtrate antigen for detection of delayed-type hypersensitivity in cryptococcosis.

Previous studies on a cryptococcal culture filtrate (CneF) antigen have shown that the antigen is useful in detecting delayed-type hypersensitivity and that it is specific for Cryptococcus. This study further defined one more parameter of specificity, showing that the CneF antigen does not elicit delayed-type hypersensitivity responses in Cryptococcus albidus-sensitized guinea pigs. When the crude CneF antigen was subjected to ultrafiltration fractionation, the skin test active components were found to be in the 50,000 or greater molecular weight range fraction. The concentrated retentates of the XM50 ultrafiltration membrane were more sensitive antigens than the crude CneF antigens. Further fractionation of the XM50 retentate using 3% acrylamide gel electrophoresis separated the antigen into two bands. One band, the P fraction, migrated only a short distance into the gel; the fraction was carbohydrate-like and did not elicit significant skin test responses in sensitized guinea pigs. The other band, G fraction, appeared with the tracking dye, was glycoprotein-like, and elicited significantly positive skin tests in sensitized guinea pigs. G fractions prepared using three different serotypes of Cryptococcus neoformans elicited similar size indurations when used in skin testing guinea pigs sensitized with either the homologous serotype isolated of C. neoformans or the heterologous serotype isolate.


Capsular K1 polysaccharide of Escherichia coli: relationship to virulence in newborn rats and resistance to phagocytosis.

The virulence of Escherichia coli strains for newborn rats was related to opsonic requirements of the strains, sensitivity to the bactericidal activity of serum, and K1 capsular polysaccharide content. K1 E. coli strains were more virulent than non-K1 strains after intraperitoneal injection in newborn rats (P less than 0.05) and were more resistant to phagocytosis than non-K1 strains when the classical complement pathway was blocked with Mg-ethyleneglycoltetraacetic acid (P less than 0.0005). Sensitivity to the bactericidal activity of serum was similar among K1 and non-K1 E. coli strains. Two groups of K1 E. coli strains were defined on the basis of opsonic requirements. Group I strains were efficiently opsonized by the alternative complement pathway, while group II strains required the classical complement pathway for opsonization. Group I strains had less detectable K1 polysaccharide in the washed whole cell fraction than group II strains (10.3 versus 18.9 microgram of K1 polysaccharide per 10(10) colony-forming units) and were less virulent than group II strains (mortality, 44 versus 77%, P less than 0.05). The K1 capsular polysaccharide appears to play an important role in determining virulence in newborn rats and opsonic requirements of these strains, but does not contribute to the sensitivity of strains to the bactericidal activity of serum.


Immunosuppressive activity of BCG: effects of adjuvant disease, lymphocyte subpopulations, and homing of thoracic duct cells in rats.

Administration of BCG by various dosage schedules suppressed adjuvant disease in rats. BCG administration produced an initial increase, followed by a depression, of the phytohemagglutinin response of purified blood lymphocytes. An increase in absolute and relative numbers of bursa-equivalent (B)-cells followed BCG administration, concurrent with a decrease in the phytohemagglutinin responsiveness. With adjuvant alone, there was a diminution in phytohemagglutinin response and an increase in number of B-cells; the latter occurred immediately after adjuvant injection and also when the generalized disease appeared. When both BCG and adjvant were present, parallel increases of phytohemagglutinin responsiveness and B-cell numbers resulted. The pattern of tissue localization of radioactively labeled thoracic duct cells from normal or BCG-treated donors given to normal, BCG-treated, adjuvant-injected, and BCG-treated + adjuvant-injected syngeneic recipients indicated significantly greater homing to the thymus and decreased localization to the bone marrow when BCG had been given to either donors or recipients. When labeled thymus cells were used, only the decreased bone marrow localization was noted. These observations suggest that the suppressive effect of BCG may be mediated through modification of the lymphocyte recirculation pattern, possibly resulting from alterations in lymphocyte recognition sites.


Endotoxin toxicity in rats with 6-sulfanilamidoindazole arthritis.

Seven oral administrations of 6-sulfanilamidoindazole (6-SAI) to 10- to 12-month-old rats sensitized the animals to endotoxin, with dosages as small as 2.5 microgram causing death in 80% of animals. Endotoxin in a dosage of 3,000 microgram was not lethal for nonmedicated control animals. 6-SAI-treated 1-month-old rats were not as sensitive to endotoxin as aged animals. The sulfonamide-induced sensitivity to endotoxin could not be passively transferred and could not be explained by blockade of the reticuloendothelial system or impairment of endotoxin detoxification. 6-SAI administration was associated with both depletion of liver glycogen and lowering of blood glucose concentration without changes in blood lactic acid concentration. Disseminated intravascular coagulation is believed to be involved in the pathogenesis of shock and death as evidenced by: (i) concomitant decreases in plasma fibrinogen concentration and elevations in fibrin degradation products after endotoxin challenge; (ii) protection against lethal actions of endotoxin by pretreatment with heparin. Treatment of 6-SAI-medicated rats with glucocorticoids before endotoxin challenge protected the animals against lethal doses of endotoxin and prevented deposition of fibrin thrombi in the glomerular capillaries.


Galactomannan antigenemia in invasive aspergillosis.

Galactomannan (GM) extracted from mycelia of Aspergillus fumigatus with cold dilute alkali reacted with antiserum specific for an antigen that circulated in invasive aspergillosis in rabbits and humans. The GM was purified by its affinity for concanavalin A and was separated from a nonantigenic glucan by gel permeation on Sephacryl S-200. The GM molecular weight of between 25,000 to 75,000 was smaller than the antigen present in infected rabbit serum which was retained by an ultrafiltration membrane that had a nominal molecular weight limit of 125,000. The ratio of galactose to mannose present in GM was 1:1.17. The serological activity of GM was stable to boiling but labile to 0.01 N HCl, implicating galactofuranose as an antigenic determinant. Analysis of purified GM by methylation-gas chromatography suggested a structure consisting of a 1 leads to 6-linked mannan backbone with oligogalactoside side chains 3 units long, terminating in galactofuranose. The presence of mannose as a side chain component was also inferred. Another antigen of A. fumigatus, which did not bind to concanavalin A, was isolated after tandem chromatography on diethylaminoethyl- and carboxymethyl-Sephadex and was identified as a galactan. The galactan inhibited the immune precipitation of GM was specific antiserum.


Immunological behavior after mycobacterial infection in selected lines of mice with high or low antibody responses.

Resistance and susceptibility to mycobacterial infection in the Biozzi high and low lines of mice which were genetically selected for their responses to heterologous erythrocytes have been found to be related to the innate ability of nonimmune macrophages to kill or inhibit the growth of the organisms during the first two weeks after infection and to their ability to mount specific and nonspecific immune responses. High antibody-producer mice were more capable of expressing cell-mediated immune parameters than low antibody-producer mice. A direct relationship was observed between the ability of bacteria (BCG vaccine) to multiply inside the reticuloendothelial system and the development of cell-mediated immunity, as measured by the delayed local reaction at the injection site, the lymphoproliferative response in the draining nodes, the tuberculin delayed-type hypersensitivity, the acquired resistance, and the adjuvant effect after BCG inoculation. In high line mice, apart from the inability of their macrophages to inhibit the early growth of bacteria, their lymphocytes in spleen and thymus were more capable of being stimulated in vitro by varying concentrations of living BCG. The data presented in this report are compatible with the hypothesis that a group of genes segregated in each line during the selective breeding controls the innate microbicidal activity.


Virulence of Escherichia coli in experimental hematogenous pyelonephritis in mice.

Differences in nephropathogenicity between Escherichia coli strains were studied by following the kinetics of the viable count in the mouse kidney during 8 h after intravenous injection. Assuming as a reference point that at zero time 0.1% of the inoculum was lodged in the kidney, we found that strains fell into three main groups with different behavior patterns: in group I, the viable count fell and remained low; in group II, the viable count first fell and then rose after 4 h, reaching the level of the reference point within 8 h; in group III, the viable count rose rapidly and remained high. The kinetics of the viable counts were also studied in blood, spleen, and liver: group I and II strains behaved similarly; only in the kidney did group II strains show higher counts than group I strains. These data suggest that group II strains are specifically virulent for the mouse kidney. Group III strains also gave higher viable counts in blood and spleen but comparable counts in the liver, suggesting that group III strains are more generally virulent. Fifty percent lethal dose measurements confirmed the conclusion that group I strains are avirulent and group III strains are the most virulent. Possible relationships between behavior pattern and serotype are discussed.


[Experience with sisomicin in pediatrics (author's transl)].

Sisomicin, an aminoglycoside antibiotic, is especially effective against Escherichia coli, Klebsiella, Enterobacter, Citrobacter, Serratia, indole-positive and indole-negative Proteus species, Pseudomonas aeruginosa, Salmonella and Staphylococcus aureus. It has a bactericidal action. Although sisomicin is similar to the other aminoglycoside antibiotics, there is not complete cross-resistance to them. Our own pharmacokinetic investigations showed that a dose of 2--3 mg/kg body weight of sisomicin twice daily is necessary in the neonatal period. Infants should be given 2.5 mg/kg body weight three times daily, and school children 1.5--20 mg/kg body weight, likewise three times daily. Excretion of sisomicin in the urine is lower in children than in adults, amounting within 24 hours to only 10--20% in newborns, and 30--40% in school-children. Sisomicin induces excretion of some enzymes in higher quantities from the tubular part of the kidneys, especially alaninaminopeptidase. A report is given on 58 patients, especially newborns and prematures, who were treated for about seven days with sisomicin. The results obtained with a wide variety of infections (such as omphalitis, aspiration of amniotic fluid with broncho-pneumonia, phlegmons of the galea, and also pyelonephritis and mucoviscidosis with pulmonary complications) can be described as good, with a success rate of 85%. On only seven occasions were insignificant transitory side-effects, such as slight increase in transaminases, toxic-allergic exanthema and pain in the region in injection, observed.