Murine Ia and human DR antigens: homology of amino-terminal sequences. Murine Ia and human DR antigens were isolated and purified by immunoprecipitation and sodium dodecyl sulfate/polyacrylamide gel electrophoresis with allo- and xenoantisera, respectively. The I-A subregion antigen consists of two chains, designated Aalpha and Abeta, with molecular weights of 35,000 and 26,000, respectively. The I-C subregion antigen likewise consists of two chains, designated Calpha and Cbeta, with molecular weights of 32,000 and 29,000, respectively. Under nonreducing conditions, the Cbeta chain migrates appreciably more rapidly on sodium dodecyl sulfate/polyacrylamide gels than the reduced Cbeta chain, reflecting the presence of an intrachain disulfide bond. The human DR antigen is also a two-chain unit and contains DRalpha and DRbeta components with molecular weights of 34,000 and 28,000, respectively. The DRbeta chain migrates more rapidly before reduction than afterward, like the murine Cbeta chain. The DRbeta and Cbeta chains are also strikingly homologous if a single amino acid shift is imposed on one of those chains. Thus, human DR antigens strongly resemble the murine I-C subregion antigens.


AB variant of infantile GM2 gangliosidosis: deficiency of a factor necessary for stimulation of hexosaminidase A-catalyzed degradation of ganglioside GM2 and glycolipid GA2. Human kidney extracts heated to 60 degrees and devoid of hexosaminidase activity (2-acetamido-2-deoxy-beta-D-glucoside acetamidodeoxyglucohydrolase EC 3.2.1.30) stimulate more than 20-fold the hexosaminidase A-catalyzed degradation of ganglioside GM2 and of glycolipid GA2, the neuronal storage compounds of GM2 gangliosidosis. The stimulating factor of this extract, which is labile at temperatures above 60 degrees, is also present in kidney extracts from patients with infantile GM2 gangliosidosis having a deficiency of hexosaminidase A (Tay-Sachs disease, variant B) and a deficiency of hexosaminidases A and B (variant 0). Evidence is presented that this factor is defective in the AB-variant of infantile GM2 gangliosidosis which is characterized by an accumulation of glycolipids GM2 and GA2 despite the fact that the degrading enzymes, hexosaminidases A and B, retain normal activity levels. Thus, variant AB is an example of a fatal lipid storage disease that is caused not by a defect of a degrading enzyme but rather by a defective factor necessary for the interaction of lipid substrates and the water-soluble hydrolase.


Aggression increase and water competition decrease in squirrel monkeys given physostigmine injections. Six adult squirrel monkeys (Samiri sciureus) were tested in pairs for social dominance in a water competition task. Dominance was defined by two methods: (1) number and direction of aggressive responses, and (2) successful water competition as assessed by latency to the water bottle, latency to accumulate 15 seconds of drinking, and total drinking duration. Monkeys were assigned to pairs on a "round robin" basis so that each monkey was paired with all other monkeys under control, saline, and three levels of physostigmine sulfate (6.25, 12.5, and 25.0 microgram/kg). Under drug conditions only one member of each pair was drugged. The 12.5 microgram/kg dose of physostigmine resulted in a significant increase in aggressive responses. Both drinking duration and general motor activity decreased with increasing dose level of drug. It was also noted that the non-drugged partners of drugged monkeys accumulated 15 seconds of drinking faster and drank more at the 25 and 12.5 microgram/kg dose levels than under control and saline conditions. Physostigmine resulted in an increase in one measure of social dominance and a decrease in the other.


SKF 525-A inhibition of hepatic monooxygenase activity in rhesus monkeys. Pretreatment of 3 male rhesus monkeys (Macaca mulatta) with SKF 525-A(20 mg/kg, i.m.) 3 h prior to to injecting 14CH3-N-antipyrine (30 mg/kg, i.v.) increased antipyrine plasma half-life (APH) 240--500%; measurements of APH made 4, 8 and 11 days after SKF 525-A treatment were also higher than control values. Increases correspond to decreased plasma levels of 3-hydroxymethylantipyrine and 4-hydroxyantipyrine and decreased rates of aldrin epoxidation in liver biopsy homogenates. Comparison of HSLC and radiometric methods for determining APH showed that both methods were capable of detecting chemically altered hepatic monooxygenase activity.


Role of glucagon in the hyperglycemic response to catecholamines in fasted baboons. The hyperglycemic activities of epinephrine (EPI) and isoproterenol (ISO) in baboons correlated with their ability to increase plasma glucagon (IRG) levels relative to insulin (IRI). EPI inhibited IRI release and produced greater increases in plasma glucose and IRG than did ISO. ISO increased plasma IRI levels more than IRG. Infusion of somatostatin blocked IRG release and inhibited hyperglycemic responses to EPI by approximately 50%. These findings indicate that, as in man, IRG release contributes significantly to the hyperglycemic effects of catecholamines in baboons. The baboon thus appears to be a suitable model for predicting effects of drugs on glucose homeostasis in humans.


Regeneration of skeletal muscle after grafting in monkeys. Twenty-five palmaris longus muscles were transplanted into the forearm (orthotopic autografts) or into the face (heterotopic autografts) in 15 Rhesus monkeys. These muscles were transplanted with or without anastomosis to a motor nerve. No significant difference was observed long-term between the grafts done with or without prior denervation of the muscle. The forearm muscle grafts without neurorrhaphies formed a static, fibrous sling which did not contract. The forearm autografts with neurorrhaphy and the grafts to the face, with or without neurorrhaphy, all developed regenerating skeletal muscle fibers and showed contractile activity.


Demonstration of actin in the fibroblasts of healing palatal wounds. The purpose of this study was to gain information which might be helpful in understanding the influence of palate surgery on abnormal maxillary growth. The finding of actin-rich cells in the granulation tissue of palatal wounds supports the hypothesis that one of the functions of those cells is contraction--and the latter subsequently leads to convergence of the wound margins and aberrations in growth of the underlying skeletal tissue. Much additional information is needed for final proof of that hypothesis. At present little is known about the initiation, regulation, or control of the proposed contractile phenomena, or the process of aberrent maxillary growth. Such information will be necessary for further development of clinical techniques that will allow normal maxillofacial growth after surgery on the palate at an early age.


[Leg ulcer caused by a monkey's bite].

[Leg ulcer caused by a monkey's bite]. 35 days following a monkey's bite, the ulcer observed showed total necrosis associated with spotted zones of necrosis due to the deep penetration of the animal's teeth. It was treated like a regular ulcer and healed within seven weeks without complications. One should keep in mind that such ulcers involve infectious episodes due to the transmission of a pathogenic human-type flora through a septic orodental milieu: Pathogenic cocci; Pasteurella, Shigella, Salmonella; Polio viruses or microbial and viral agents of the animal itself, which can cause lethal diseases such as: Yellow fever, Marburg's disease, Rabies.


Enhanced intestinal lymph formation during fat absorption: the importance of triglyceride hydrolysis. The effect of intraduodenal administration of fats was studied in the rat to define the mechanisms responsible for the substantial increase in intestinal lymph flow and protein transport which follows fat ingestion. Triglyceride in the intestinal lumen, protected from hydrolysis, does not appear to enhance intestinal lymph production. Giving both long- and medium-chain fatty acids, however, causes intestinal lymph flow and protein transport to increase in a manner similar to that found after giving triglyceride which is allowed to undergo hydrolysis. Bile by itself does not seem to be responsible for the phenomenon.


The validity of the approximation method "area/perimeter" as applied to 8 and 25-MV photon beams. The approximation method of area/perimeter for different portal sizes used in practice for finding the equivalent squares of a rectangular field has been tested on a 35-MV Clinac Linear Accelerator for the purpose of treatment planning. Data were obtained for 8 and 25-MV x-rays at depths of maximum build-up regions, and at 10 cm depth below the surface of a suitable water phantom. Maximum variation of dose calculation from the above method is less than 1.6% for routine clinical usage while less than 3.2% for extreme rectangular field sizes. Dose variation improves as the depth in phantom increases.


Ineffectiveness of prophylactic intrathecal methylprednisolone in myelography with aqueous media. The effectiveness of prophylactic intrathecal methylprednisone (MP) in preventing arachnoiditis from iocarmate myelography was studied. Monkeys were injected intrathecally with locarmate, MP, or a combination of the two. Twelve weeks later the severity of arachnoiditis was determined in each animal. There was no significant prophylactic effect of intrathecal MP on arachnoiditis from locarmate. Some arachnoiditis was found in control animals treated with MP alone.


[Acute non-lithiasic cholecystitis].

[Acute non-lithiasic cholecystitis]. The authors present 7 cases of acute non-lithiasic cholecystitis, representing 1.5% of the total number of acute cholecystitis cases in their personal statistics. The determining factor of the acute inflammatory process of the gallbladder was represented in 6 of the cases by non-lithiasic obstruction of the cystic duct. In the last case there was acute segmental cholecystitis secondary to a gallbladder septum, probably of congenital origin. Clinical and anatomo-pathological aspects are discussed, of acute non-lithiasic cholecystites, the origin of which is located in the diverticular apparatus of the biliary pathways. From the therapeutical viewpoint cholecystectomy is the most indicated attitude in such cases.


The effect of ethanol on the metabolism of chronically-administered l-alpha-acetylmethadol (LAAM) in the rhesus monkey. The metabolism of 3H-LAAM was studied in monkeys treated acutely and chronically with 2 mg/kg LAAM via intragastric catheter three times weekly. From the 15th week, 250 mg/kg ethanol every 2 hours (3g/kg/day) was also administered. This dose of ethanol did not impair the metabolism of LAAM in monkeys.


[Endocranial complications in hemopathies. Merits of computerized axial tomography (CAT) (author's transl)]. On the basis of 88 patients examined with CAT, the writers consider the merits of this method of investigation in a study of endocranial complications in blood diseases. Interest in CAT is related in great part to its innoccuousness (making it possible to extend its application to often seriously ill patients) and the precision of the images which are obtained. This precision makes it possible to visualize lesions which at times can not be detected with other methods. A great part of present knowledge on endocranial complications in blood diseases is based on autopsy findings. With CAT, there is definitely the possibility of performing anatomical studies during the patient's lifetime, thus providing a better understanding of the characteristics and frequency of these complications. This is already made clear by this preliminary study, particularly in the area of acute leukemia and Hodgkin's disease.


[Preoperative radiotherapy and curative radiotherapy without mastectomy in carcinoma of the breast (author's transl)]. The aim of preoperative radiotherapy of mastocarcinoma is to reduce the size of the tumor to enable radical surgery in the third stage. This method is preferable to postoperative radiotherapy for anatomical and radiobiological reasons. It has not yet been possible to prove beyond doubt that preoperative radiotherapy also has a favourable effect on symptom-free survival. Curative radiotherapy without mastectomy is recommended for tumors of 2 - 3 cm size which are mobile, in small and medium-sized breasts, if the axilla is free or has only 1 - 2 mobile lymphatic nodes. Tumorectomy must be followed by surgical revision of the axillary region. High-voltage equipment is mandotory for radiotherapy. Results obtained to date are in no way inferior to those of mastectomy. Local control rate is 90-100%. The cosmetic result is usually excellent.


[Chemotherapy as an adjuvant treatment in breast cancer (author's transl)]. Prolonged L-Phenylalanin mustard Administration and combination chemotherapy as on adjuvant to mastectomy in the management of patients with primary breast cancer and pathologically axillary nods. Lead to improvement of results. L-PAM and CMF have been demonstrated to be effective in the treatment of women with primary breast cancer, particularly those who are premenopausal. In our trial no improvement of results could be observed by administration of Chlorambucil, Cyclophosphamide and 5-FU. Experience with both single-drug and combination chemotherapy has shown that the benefit of these treatments, as judged by the disease-free intervall, is restricted largely to premenopausal patients. This suggest that the therapeutic effect is due, at least to cytotoxic suppression of endocrine function. Therefore further prospective trial in this field are necessary.


[A contribution to radiological diagnostics of xanthogranulomatous pyelonephritis (author's transl)]. Pyelonephritis xanthogranulomatosa is a peculiar manifestation of destructive interstitial nephritis which can be differentiated from other types of pyonephrosis exclusively by microscopic examination. Clinical symptomatology often points towards the existence of a renal tumor. The author analyses the clinical and radiological symptomatology of seven patients treated by him. The angiographic picture presented by pyelonephritis xanthogranulomatosa differs according to the character and extension of the inflammatory tissue lesion. The angiographic picture of the circumscribed and diffuse granular proliferative types resembles that of a poorly vascularised malignoma, whereas the destructive type resembles hydropyonephrosis.


[Ossification of the coracoclavicular ligaments (author's transl)]. Joint-like connections between clavicula and coracoid process are rarely diagnosed. In routine radiological examinations of the thorax, the authors found a posttraumatic ossification of the coracoclavicular ligament in four patients. This phenomenon can occasionally be the cause of shoulder pain. The anatomy and pathogenesis are described, with x-ray film examples, on the basis of four cases.


Lactose malabsorption after bypass operation for obesity. After bypass operation for obesity the remaining lactose-hydrolyzing capacity of the functioning shunt is very low, especially if the shunt is constructed from a shorter jejunal and a longer ileal segment. In most cases a temporary decrease in the lactase activity of the jejunal part of the shunt occurs during the first postoperative months. In the present study lactose provoked or aggravated diarrhoea and other symptoms in 20 of 33 shunt-operated patients, and 10 patients reported milk intolerance postoperatively. Oral glucose tolerance tests indicated that the lactase activity was rate limiting for lactose absorption postoperatively.


Determination of glomerular filtration rate in advanced renal insufficiency. The glomerular filtration rate (GFR) has been determined in 17 patients with advanced renal insufficiency (GFR less than 15 ml/min) by different clearance techniques using creatinine, inulin and 51Cr-EDTA as filtration markers. With renal inulin clearance as reference method for GFR, endogenous renal creatinine clearance overestimated GFR by an average of 30%. Renal clearance of 51Cr-EDTA and inulin were closely correlated and thus 51Cr-EDTA is a suitable GFR marker even at low filtration rates. However, it was found that the plasma clearance of 51Cr-EDTA overestimated the GFR often by more than 100% in the range 2.6--11.2 ml/min. Renal clearance measured during 24 h was lower than 4 h renal clearance with the patient well hydrated and resting in bed. It is concluded that the precise measurement of low glomerular filtration rates requires the use of renal clearance techniques. Four-hour 51Cr-EDTA renal clearance is a suitable method for measuring and following the development of renal function in advanced renal insufficiency.


Lead poisoning treated with haemodialysis. A 27-year-old woman ingested as an abortifacient 40 grams of litharge (PbO). She was treated with EDTA infusion and haemodialysis. The half-life of lead in the blood was 9 hours during combined haemodialysis and EDTA infusion and 96 hours when EDTA was given alone. Signs and symptoms attributable to acute lead intoxication were mild.


Tuberculosis infection and the repeat test. Effect in BCG-vaccinated guinea pigs. Groups of BCG-vaccinated guinea pigs were infected with Mycobacterium tuberculosis and the course of tuberculin reactions before and after was studied. The infection led to reactions that were accelerated as if repeated tests had been given, but were larger than in such cases. Acceleration of reactions due to previous testing was present 1 week, but not 4 weeks after the start of the infection. Possibly, in cellular immunity in general, absence of the repeat test effect may be interpreted as a sign that specific antigen stimulation is taking place, and diagnostic tests could be constructed on this basis.


[Pseudothrombopenia (proceedings)].

[Pseudothrombopenia (proceedings)]. Two cases of spurious thrombocytopenia are reported, one induced by platelet satellitism and the other by platelet aggregation. These phenomena occur in vitro, only in the presence of EDTA and are linked with the presence of IgG in the patients' sera.


[Mobility measurements and histological studies on endosseous titanium implants under masticatory stress in the mandibles of adult macacas]. Linkow blades were implanted into 5 adult monkeys. Measurements of mobility showed that two weeks after implantation the implant mobility was below the mobility of the control teeth. Fixed bridgework was inserted and after 8-14 months the mobility of implant and control teeth were alike. Inspite of this favourable result, the mobility of the implants, after removal of the bridgework, was higher than that of the controls and of normal values. The histological picture explains this, since all implants showed a sheath of connective tissue which was 5-10 times the dimension of the normal desmodontal space. Inflammatory processes and epithelial growth into depth were observed inspite of the absence of clinical symptoms. There was bony growth through the windows of the implants, but only on two implants could bone sheathing be observed. The conclusion is that Linkow implants should not be used alone for the anchoring of bridgework but only as link supports between long bridge spans or as distal extension of shortened tooth arches. Also, it can be used for the retention of a Dolder bar in mucosa supported dentures.


Processes controlling arm movements in monkeys. The experiments identify some of the processes underlying arm movements in rhesus monkeys. Three monkeys were trained to point to a target with the hand and forearm and to hold that position for about 1 second to obtain a reward. Forearm movements were performed without sight of the arm before and after bilateral dorsal rhizotomy. In both intact and deafferented animals, we unexpectedly displaced the forearm prior to movement initiation and observed that the arm moved accurately to the target. These results are relevant to the question of what is being controlled by motor commands. The controlled variable appears to be an equilibrium point between agonist and antagonist muscles. The findings suggest that the feedback system plays a major role in updating and adjusting the central programs subserving the execution of learned motor patterns.


Vaccination of experimental monkeys against Plasmodium falciparum: a possible safe adjuvant. Owl monkeys (Aotus trivirgatus griseimembra) were effectively immunized against a human malaria parasite, Plasmodium falciparum. Two injections of antigen, primarily mature segmenters with fully developed merozoites, mixed with adjuvant (6-O-stearoyl-N-acetylmuramyl-L-alanyl-D-isoglutamine and liposomes) were administered intramuscularly at a 4-week interval. Approximately 2 weeks after the second vaccination, the monkeys were challenged with the homologous strain of P. falciparum. All immunized monkeys survived the challenge. The substitution of Freund's complete adjuvant is an encouraging step toward the development of an effective and safe vaccine for human malaria.


Antibody diversity.

Antibody diversity. Three important aspects of immunoglobulin gene organization and structure have emerged from studies of cloned immunoglobulin kappa chain genes. (i) Multiple variable genes are encoded separately in the genome of both immunoglobulin-producing and uncommitted (embryonic) cells, thereby establishing the evolutionary base for generating immunoglobulin diversity. (ii) These genes exist as many small, closely related families (subgroups) that share close sequence homology largely within their own subgroup. (iii) Comparison of two cloned variable gene segments derived from a single subgroup reveals a feature of their structure that distinguishes them from fixed genes (that is, globin genes) and provides, through extensive surrounding sequence homology, a large target for intergenic recombination. This last observation suggests that a simple recombination mechanism may account for their genetic instability in both germ line and somatic cells.


Neural lateralization of species-specific vocalizations by Japanese macaques (Macaca fuscata). Five Japanese macaques and five other Old World monkeys were trained to discriminate among field-recorded Japanese macaque vocalizations. One task required discrimination of a communicatively relevant acoustic feature ("peak"), and a second required discrimination of an orthogonal feature of the same vocalizations ("pitch"). The Japanese animals more proficiently discriminated the peak feature when stimuli were presented to the right ear (primarily left cerebral hemisphere), as opposed to the left ear (primarily right hemisphere). In discriminating the pitch feature, the Japanese animals either showed (i) a left-ear processing advantage or (ii) no ear advantage. The comparison animals, with one exception, showed no ear advantage in processing either feature of the vocalizations. The results suggest that Japanese macaques engage left-hemisphere processors for the analysis of communicatively significant sounds that are analogous to the lateralized mechanisms used by humans listening to speech.


Productivity of ospreys in Connecticut--Long Island increases as DDE residues decline. Nesting success of ospreys (Pandion haliaetus) breeding in the Connecticut--Long Island area has increased since 1973 and is now approaching the levels recorded prior to the 1950's. Simultaneously, DDE and dieldrin residues have declined in unhatched eggs. Levels of polychorinated biphenyls have shown no changes over the period 1969 to 1976. The increase in productivity is attributed primarily to lower levels of DDE contamination. Detrimental effects in the past on ospreys in the Connecticut River estuary are attributed to local contamination with dieldrin.


Induced abortion.

Induced abortion. The morbidity and mortality of illegal abortion are briefly discussed with reference to South Africa, where it seems that 1 in every 8-10 pregnant women deals with an unwanted pregnancy in this way. A review of the literature regarding legal abortion has been undertaken, detailing mortality and morbidity with various methods in many countries. The medical problems resulting from a permissive abortion policy have been highlighted, and an attempt has been made to define the place of medically induced abortion in health services, as it is understood by countries with extensive experience in this field. It is hoped that this contribution will help to define the real issues in the current abortion debate in South Africa.


Cellular immune function in marasmic and underweight infants with prolonged diarrhoea. Lymphocytes from 10 marasmic and underweight infants with prolonged diarrhoea responded normally to phytohaemagglutinin stimulation. There was no gross deficiency in the total number of lymphocytes, serum immunoglobulins, or serum C3. Defective cellular immunity does not appear to play a major role in the pathogenesis of diarrhoea in these children.


Postoperative ileus: a colonic problem?

Postoperative ileus: a colonic problem? Postoperative electromechanical activity of the gastric antrum, small bowel, right colon, and sigmoid colon was recorded in stumptail monkeys in response to retroperitoneal dissection and transient clamping of the renal pedicle. Bipolar silver electrodes and extraluminal bonded strain gauge transducers were used to record slow-wave and spike discharges and contractions of intestinal smooth muscle. After operation myoelectric activity was decreased transiently in the antrum and for only a few hours in the small bowel. Right colon contractile activity was decreased significantly for 24 hours and that of the sigmoid colon for 72 hours. Postoperative inhibition of bowel motility appears to be most profound and persistent in the colon.


Genetic control of the interferon system. Now that some studies have been done on the genetics of the human interferon system, it would appear that the primary constraint to interferon research imposed by the lack of human interferon can be overcome as indicated herein with mutagenized human cell lines containing multiple copies of chromosome 5. Purified interferon from these mutants has allowed us to label it radioactively. However the amount required for a complete amino acid and sugar sequence of human interferon is still very large but within the realm of possibility when the use of these mutant cell lines is considered (23, 24).


Inhibition of interferon production by retinoic acid (vitamin A acid). The production of interferon was employed to study the site and mechanism of action of retinoic acid (vitamin A). The data is consistent with a site of action of retinoic acid at the gene level, since it appears that interferon production is suppressed at the transcriptional step by a retinoic acid-induced protein. Evidence for this is as follows: (a) The effect of retinoic acid is probably on an early cellular function associated with interferon production rather than an effect on the inducer (virus or poly I:poly c). (b) The suppression of interferon production by retinoic acid is blocked by cycloheximide indicating a newly synthesized protein (repressor) mediates the suppression. (c) When allowances are made for the time required for the synthesis of the retinoic acid-induced protein, the time course of retinoic acid suppression of interferon production is superimposable on the time course of actinomycin D suppression since the slopes are very similar. These data provide the first evidence for transcriptional control of a specific protein (interferon) by retinoic acid. Additionally, they support the existence of transcriptional control of interferon production after inducer addition.


Induction of the interferon protein in the animal: general considerations. Serum interferon is largely produced by B and T lymphocytes and macrophages, depending on the inducer. Lymphocyte interferon is different from macrophage and fibroblast interferon. There are two alleles controlling mouse interferon production. A protein possibly related to interferon may cause hyporeactivity. Our work was supported by NIH Grant 5-RO1-02953.


Possible associations between HLA antigens and the immune responsiveness to attenuated rubella vaccine. We investigated the possible associations between HLA antigens and the antibody response patterns during a clinical trial with a live attenuated rubella vaccine in 172 Japanese schoolgirls. Those vaccine recipients were divided into three groups, 42 low responders, 102 intermediate responders and 28 high responders, according to convalescent-phase hemagglutination-inhibiting antibody titers. High frequencies of HLA--A11 and HLA--B15 in high responders and HLA--Aw24 and HLA--B5 in low responders were found. Among HLA--A antigens HLA--A11 had the highest geometric mean HI antibody titer and HLA--Aw24 had the lowest. Among HLA--B antigens HLA--B15 had the highest and HLA--B5 had the lowest. Strong linkage disequilibrium was found between HLA--A11 and HLA--B15 in high responders and between HLA--Aw24 and HLA--B5 in low responders. These results suggest that there may be a relationship between HLA antigens and specific immune response genes in man as well as in other vertebrates.


Changes of cell cycle affecting cidal effect of triethylene thiophosphoramide on cultured human bladder carcinoma cells. The cell cycle of the human bladder carcinoma cell line (MGH-U1 cells) was prolonged immediately after low dose of triethylene thiophosphoramide (Thio Tepa). Although the elongated cell cycle time seemed to return to the control level 48 hr following the treatment, there still remained postsynthetic accumulation and postmitotic diminution in the cell population and the cells were insensitive to high dose of the agent. The present results suggest that Thio Tepa is less effective for G2 cells, but more effective for G1 or S cells.


[Radiobiological bases and clinical aspects of radiation therapy with heavy particles (author's transl)]. Our present radiobiological knowledge gives rise to the hope of an amelioration of the local results in radiation therapy by means of new categories of radiations. Neutrons are clinically used to a larger extent already, protons occasionally. The physical characteristics of pi-meson beams are especially favorable. Even considerable ameliorations, however, of the percentage of local healing would increase the survival rate by some few per-cent only.


[Pharmacologic analysis of the stimulating effect of serotonin on proliferation in cell cultures]. Serotonin (10-7 M) stimulated cell proliferation in the primary cultures of mouse and human embryonic fibroblasts as well as in mouse L-cells and in monkey kidney cells (MA). The stimulatory effect is completely prevented by pretreatment of cultured cells with serotonin antagonists of tipindole, morphine or cyproheptadine type which do not affect cell proliferation. It is therefore assumed that the stimulatory action of serotonin on these cells is realized via specific serotonin receptors.


[Effect of anesthesia on the results of the overall treatment of breast cancer]. The authors have studied the 5 years' survival of 403 cases of mammary gland cancer without development of recurrences and metastases. In 205 out of them ether-nitrous oxide anesthesia and in 198 fluoroxene-nitrous oxide anesthesia were applied. Postoperative Thiophosphamide (Thio-TEP) was administered to all the patients. The main factors characterizing the compared groups were identical in both of them. It has been proved that 5 years survival of mammary gland cancer cases rises when the patients undergo a complex treatment, and during the operation they are subjected to fluoroxene and not to ether.


[Size of pellets and individual properties of boar semen in relation to the number of motile spermatozoa following defrosting]. The second fraction of the ejaculate of five boars of the Landrace breed was used for the experiments. Non-diluted semen was kept in hydrogen atmosphere for three hours at + 5 degrees C; then it was frozen in solid carbon dioxide into 0.1 ml, 2.0 ml, and 5.0 ml pellets and in liquid nitrogen vapours into the shape of disk or cylinder 5 ml in volume. The pellets were de-frozen on a teflon pan at 42 degrees C without addition of any other diluent. Ejaculate in which at least 25% of the spermatozoa were motile was considered to be usable. No statistically significant difference was found between the motility of spermatozoa frozen in the 0.1 ml and 2.0 ml pellets, but it can be said that on an average the motility of the spermatozoa was lower in larger pellets; this could be observed mainly in the pellets with the volume of 5.0 ml. Large differences were revealed in the freezability of the semen of different boars, irrespective of the size of the frozen pellet. Insemination of 38 gilts with semen from 2.0 ml pellets gave a 57,8 % concentration rate (22 gilts) and the average litter size was 8.25 piglets.


[Effect of bilateral orchidectomy on serum testosterone levels in boars]. Bilateral orchidectomy caused a marked decrease in the concentration of testosterone in the bood plasma of boars already in the early post-castration stage of three hours after the operation. In the period from 3 to 207 days after castration, the pre-operation levels of plasma testosterone decrease by 84.4%, on an average. Adreno-cortical compensation of the concentration of the hormone did not occur in the post-castration period. The conclusion is that the levels of testosterone in the blood of boars are mostly of testicular origin and can be considered as the main criterion of the incretion function of the testes. The discussion concerns the practical importance of the determination of testosterone levels in the blood of boars for clinical evaluation of the hormonal function of the testes and for diagnostics of incretion hypogonodism.


[Serum testosterone levels and response to administration of chorionic gonadotropins in bovine hermaphrodites]. The concentration of testosterone in the blood and its response to administration of chorionic gonadotropin was examined in two bovine intersexes (testicular pseudo-hermaphroditism) and two control bulls without disorders in sexual differentiation. The levels of testosterone in the blood were markedly lower in both intersexes than in the control bulls. Both intersexes, unlike the control bulls, also showed a response of the blood testosterone level to the administration of chorionic gonadotropin. The conclusion is that in the given type of intersexuality the incretion function of the testes is mardedly deficient as to androgen production and functional hormonal reserve.


[Isolation and identification of Clamydia psittaci as the pathogen in enzootic abortion of sheep in eastern Slovakia]. Abortions in ewes occurred on a large scale in three localities in eastern Slovakia. Antibodies to the group-type ornithosis antigen were detected in titres of 1 : 128 to 1 : 2048 in the aborting ewes in the mentioned localities. Suspensions were prepared from the four samples of material, obtained either from the placentae and afterbirths of the aborting ewes or from the tissues of the aborted foetuses. Seven-day old yolk sacs of chicken embryos were infected with these suspensions. Four strains of Chlamydia psittaci were isolated and designated EPO-A2-uterus, EPO-B1 aborted foetus-lung, EPO-B2-aborted foetus-lung, EPO-B3-aborted foetus-spleen. Differential diagnosis eliminated some bacteria, toxoplasma, and parainfluenza-3 virus as possible agents responsible for the abortions.


[Effectiveness of cambendazole (Bonlam paste) in sheep invaded by tramatodes (Dicrocoelium dendriticum)]. Tests were performed to examine the effectiveness of a new form of cambendazole (Bonlam paste, produced by MSD, USA) in sheep, naturally invaded by the trematodes Dicrocoelium dendriticum. Single administration of 25 mg per 1 kg live weight induced a rapid decrease in the number of the excreted eggs of D. dendriticum, as counted during ovoscopic examination seven, ten, and fifteen days after administration of the chemical. The intenseffectiveness of the preparation, determined post mortem, was 95.1%. Despite this, several dicrocoelia with normal motility and with no changes on the cuticle or internal organs were found in all the animals dissected. The animals tolerated the treatment with no signs of side-effects.


[Immunological aspects of Marek's disease in chickens with maternal antibodies]. The dynamics of the production of immunoprecipitation antibodies to Marek's disease virus was studied in the serum of chickens with maternal antibodies in relation to the occurrence of the immunoprecipitation antigens of Marek's disease virus in feather follicles. One-day-old chickens were infected by the contact method with Marek's disease virus. The first occurrence of immunoprecipitation antigen was detected on the 14th day after infection and this occurrence persisted throughout the experiment, i. e. until the 112th day after infection. The antibodies were first detected the 28th day after infection and their titre kept rising until the 98th day after infection. Immunoprecipitation antibodies and antigens of Marek's disease virus were detected in some tumorously changed kidneys. Immunoelectrophoretic examination revealed in the same kidneys immunoglobulins of the class IgY, IgA and beta-globulin. The slowest-migrating fraction of IgY, together with IgA, beta-globulin and C-reactive protein were detected in the skin extracts from infected poultry. Indirect haemagglutination enabled the detection of the presence of haemagglutination antibodies in rabbit immunoglobulin to the skin antigen of Marek's disease virus, and in avian immunoglobulin to the same virus. Haemagglutination antigen was revealed in the extract from tumorously changed kidneys.


[Weight increase following administration of insulin to rats with reduced feed intake]. Young male Wistar rats, adapted to single daily administration of feed (from 8.00 to 10.00 a.m.) were studied for the intake of feed, weight gains, and feed consumption per weight gain unit after administration of Superdep insulin in the dose of 10 u kg-1 of body weight. The time limitation of access to food reduced the over-all intake of feed and weight gains and adversely affected the utilization of feed, expressed as consumption per 1 g of weight gain. The administration of insulin partly compensated the adverse effect of the reduced feed intake; this was manifested by better utilization of feeds by the experimental group, in comparison with the control animals.


[Effectiveness of ftorafur and hexamethylmelamine in advanced breast cancer]. In 7 oncological institutions of the Soviet Union a correlation was made between the efficacy of fluorofur, hexamethylmelanin and their combination for advanced cancer of the mammary gland in 136 patients. The therapeutic effect was estimated in 104 patients. Fluorofur yielded a considerable tumor regression (more than 50%) in 14 of 36 patients (40%), the duration of the remission in effectively treated patients was 2--5 months. Hexamethylmelanin induced a therapeutic effect in 18 of 37 patients (48%), the regression being complete in 6 patients (16%), its duration was 2--7 months. The combination of these drugs proved to be of an insignificant effect, the therapeutic effect was obtained in 5 of 31 patients (16%), the remission lasted for 1.5--5 months. The fluorofur therapy is rarely accompanied with adverse side effects (leucopenia--in 17%), while with hexamethylmelanin the incidence of leucopenia was 46%, a combination of the drugs reducing it up to 17%. Hexamethylmelanin combined with fluorofur was tolerated much more poorly (vomiting). Fluorofur and hexamethylmelanin are effective drugs for treatment of patients with advanced breast cancer.


The presence of active C1 (C-1) on peripheral human lymphocytes. We have shown that the first component of complement C1 is present in an active form on the surface of washed human peripheral lymphocytes but not on platelets or erythrocytes. This active C1 (C-1) was detected by its ability to transfer to sensitized cells carrying C4, i.e., EAC4, forming EAC-1,4. Active C1 was also able to consume C4. Treatment of these lymphocytes with 0.02 M EDTA removed C-1. EDTA-treated lymphocytes were able to bind exogenous purified human C-1. Comparative studies with sentized erythrocytes (EA) and EDTA treated lymphocytes showed that although fewer molecules of exogenous C1 could bind to the EDTA-treated lymphocytes than to EA, the consumption of C4 by C-1 bound to lymphocytes was significantly higher than that observed with EAC-1. When lymphocytes obtained from 2 patients with chronic lymphocytic leukemia and hypocomplementemia were tested, the release of C1, the C4 consumption and the binding of C-1 to EDTA-treated cells were highly inefficient.


Prevalence of HBeAg and anti-HBe in blood donors with HBs antigenemia. 250 HBsAg-positive sera from apparently healthy blood donors were tested for HBeAg and anti-HBe by immunodiffusion(ID) and counterelectrophoresis(CEP). HBeAg was identified in 2.4% and anti-HBe in 34% of the sera. After serum concentration a larger rate of both HBeAg and anti-HBe could be detected by ID. There was no difference in sensitivity between ID and CEP techniques in the detection of the e-system. There was no correlation between HBeAg or anti-HBe and HBsAg subtypes. Generally sera containing HBeAg showed higher titers of HBsAg in complement fixation.


Purification of hepatitis B surface antigen by affinity chromatography. Hepatitis B surface antigen (HBsAg), was purified from plasma by affinity chromatography on matrix-bound sulphated carbohydrates such as heparin or dextran sulphate. Further purification by precipitation with polyethylene glycol (PEG) 6000 resulted in a highly purified HBsAg preparation. The overall recovery amounted to about 70% of the total antigen content of the starting plasma. Electron microscopic data revealed mainly 22 nm spherical particles accompanied by few or no filaments. The process is simple, rapid and lends itself readily to large-scale applications.


Serological, immunochemical and immuoncytological properties of granulocyte antibodies. The immunochemical characteristics of granulocyte allo- and autoantibodies can easily be studied with indirect immunofluorescence techniques. The correlation of these properties with the serological behavior of the antibodies and the clinical condition in which they were found was studied. The investigations included the determination of immunoglobulin class, subclass, light-chain composition, as well as optimal temperature of activity. Complement fixation in vitro was also investigated. Striking differences were found between IgG and IgM antibodies in their serological properties, especially in the granulocytotoxicity and leukoagglutination tests at different temperatures. Differences in light-chain and IgG subclass compositions of the antibodies did not seem to play a role. In studying the distribution of granulocyte-specific antigens on normal granulocytes and granulocytes in all stages of development, it was found that, while NA1, NA2 and ND1 are present in the same quantity on the cells of an individual, the NB1 antigen is unequally distributed, both over ripe granulocytes and granulocyte precursors.


Association between HLA and red cell antigens. VII. Survival studies of incompatible red blood cells in a patient with HLA-associated haemagglutinins. Serum from a multitransfused patient with bladder papilloma was examined. The serum contained anti-Kell as well as polyspecific, cytotoxic HLA antibodies and HLA-associated haemagglutinins. The HLA haemagglutinins were demonstrated by the Auto Analyzer method and reacted predominantly with red blood cells (RBC) from HLA-A28-positive individuals, and were distinctly weaker with all other test cells. Survival studies with 51Cr-RBC from an HLA-A28-positive, Kell-negative donor with strong A28 RBC reactivity were performed. A small component of the RBC showed clearly reduced survival time, while the main component showed nearly normal survival.


Relationship between maternal anti-D levels, fetal phenotype and haemolytic disease of the newborn. It has been demonstrated that a lower level of maternal anti-D is required to produce the same symptoms of haemolytic disease of the newborn in R2r infants than in R1r infants. This difference could be explained by postulating that the higher site density of R2r cells is associated with an increased association constant and thus will bind sufficiently more anti-D. The level at which amniocentesis should be considered has been calculated to be 1.44 microgram anti-D/cm3 of maternal serum for R1r infants and 0.73 for R2r infants.


[The antibacterial activity of diphenhydramine (author's transl)]. Diphenhydramine (N,N-dimethyl-2-diphenyl-methoxy-ethylamine) [N,N-dimethyl-2-(diphenylmethoxy)-ethylamin] (DPH) is a well-known local anaesthetic and an antagonist of both acetylcholine and histamine. We tested its antibacterial efficacy by means of three different investigation methods, i.e. microcalorimetry, continuous density measurement and the microdilution test to find out the MIC of this compound against various bacteria. DPH inhibits the growth of E. coli and Klebsiella strains at a concentration of 0.18% of Staphylococcus aureus haemolyticus at 0.37%, of Pseudomonas aeruginosa at 0.75% and Streptococcus faecalis strains at 1.5%. The results of microcalorimetry and continous density measurements showed that the addition of 0.2% DPH to a broth with growing bacteria interferes very quickly with cell metabolism and stops further reproduction. The microcalorimetric findings were carried out with Staphylococcus aureus haemolyticus strain No. 13,665 an- the E. coli strain No. 3579.


[Dynamics of the appearance and disappearance of S-antigen in cells abortively of lytically infected with OV40 virus]. A highly sensitive mized hemadsorption test and blocking of it were used to study the size of the S-antigen-inducing dose of OV40 virus and the dynamics of the appearance of S-antigen under conditions of abortive and lytic infection. The data on the loss and persistence of this antigen in stationary and multiplying cultures of hamster and monkey cell are presented. S-antigen could be detected in OV40 virus-infected hamster and monkey cells long before their morphological transformation. No antigen was found upon passages of the infected hamster cells but it persisted in non-multiplying infected hamster cells as well as in productivity infected green monkey cells, irrespective of the number of passages, up to the onset cytopathic effect.


The fate of brominated soya oil in the animal body. 1. The bromine-containing material accumulated in tissues of rats dosed with brominated soya oil gradually decreased when the oil was no longer administered. 2. G.l.c. of the transesterified ether extracts of tissues from dosed animals showed 9,10-dibromostearic acid in extracts from fat, heart, kidney, liver and muscle with much smaller amounts of 9,10,12,13-tetrabromostearic acid in all extracts except that from liver. 3. An ether-soluble bromine-containing metabolite excreted by rats dosed with brominated soya oil was tentatively identified as 5,6-dibromosebacic acid. 4. The same metabolite was excreted by rabbits, rats and marmosets dosed with 9,10-dibromostearic acid.


Protein-calorie malnutrition (PCM) in Egypt immunological changes of salivary protein in PCM. Whole saliva protein as well as the separated protein components were estimated in normal and malnourished Egyptian infants and young children. In normal, 8 protein components (Albumin, alpha1-antitrypsin, haptoglobin, beta2-lipoprotein, transferrin, IgA, IgM and IgG) were detected, while in PCM cases two more components (prealbumin and alpha2-macroglobulin) were found. The results also showed that the level of salivary protein components are markedly increased in edematous cases. In non-edeomatous ones, the level of these constituents are slightly increased in 3rd marasmus, but diminished in 2nd grade. It is concluded that the elevation of protein components in saliva of edematous cases could be a result of severe glandular tissue involvement as compared to controls and non-edematous cases. The value of IgA immunoglobulin as specific antibody originated from blood plasma and/or salivary glands may be used to reflect the extent of tissue affection in salivary glands of malnourished cases.


[New strategy in the fight against malaria]. The initial spectacular success of the malaria eradication programme which has been performed since 1955 did not save from severe reactions. These reactions were the cause for a more realistic estimation of the situation. The at present necessary activities in the combat against malaria have to be realized flexibly and in the comprehensive range of the national health planning taking into consideration the epidemiological conditions, the financial and personal possibilities of the country concerned as well as the total social-economical structure. Particular problems result from the increasing resistence of the malaria vectors against insecticides and the pathogenic agents against chemotherapeutics. In the development of new methods of combat special consideration must be ascribed to their efficiency not only, but also to their economy. The at present forcedly performed malaria research is expected to give not only a better understanding of the pathogenesis of the disease, but also new possibilities of prevention.


[Liver transplantation in animal experiments]. Liver transplantation has extraordinarily complicated technical, biochemical, and pathophysiological aspect. The present study on experimental liver transplantation is a survey on essential parts of this vast problem. Particularly in the fields of biochemistry and immunology, our knowledges must be extended before an improvement of the results can be achieved. Results from experiments of the authors themselves agree with experiences, complications, and survivals after hepatic transplantation reported in literature.


Ultrastructure of sensory nerve terminals in the penis in green monkey (Cercopithecus aethiops sabaeus). Tje paper describes the ultrastructure of axons in the endings of various types from the corium in the glans penis in green monkey. In the Meissner's endings the axons are mostly completely enveloped in the plasma of Schwann cells. They contain numerous mitochondria which are partially vacuolated or are quite converted into vacuoles. Next, there are pseudomyelinated figures, light vesicles and further organelles. In the papillar simple glomerular endings with accumulation of Schwann cells there are axons irregular in shape, eccentrically placed in the plasma of Schwann cell, rounded smaller axons either completely or partially surrounded by the plasma of Schwann cell and finally axons with a concentric system of lamellae up to four in number, In the complicated glomerular endings the axons vary in appearance and are enveloped in one to five lamellae of Schwann cells, which is typical of those formations. About some of these systems there is a sign of a capsule formed by an elongated lamella probably of the perineurium. When the axons are not enveloped in the plasma of Schwann cell, they are covered by the basement membrane. In close neighbourhood of the epidermis so-called free endings forming groups were found. The plasma of Schwann cell covers them either partially or completely or it again forms around them a lamellar system amounting up to four layers. It is noticeable that these axons are very poor in organelles. A comparison of the simple sensory corpuscles in the nose skin in hedgehog, the funtional properties of Meissner's endings and the simple corpuscles results in the view that the complexes having a larger amount of lamellae correspond to an extent to the simple sensory corpuscles ant that the Meissner's endings and the complicated glomerules are probably a morphological and functional equivalent of simple sensory corpuscles in the non-primate mammals and that the gloverular endings may also be the first (developmental) stage of the simple sensory corpuscles.


[Diaplacental passage of Sarcocystis antibodies in man and rats (author's transl)]. Sera of babies up to the age of 3 months were tested for Sarcocystis antibodies using the indirect immunofluorescence test (IIFT). In addition titre of the Sarcocystis antibody level of litters born to serologic Sarcocystis positive mother rats was compared to those of their mothers. The results are as follows: 1. 45 (= 14.6%) out of 308 sera from babies up to the age of 3 months reacted positively in the Sarcocystis antibody test. 2. 28 (= 62.2%) out of the 45 positive sera were from babies up to 2 weeks old, 13 (= 28.9%) were from babies older than 2 weeks but not more than 4 weeks old, and 2 (=4.4%) each were from babies whose ages ranged from 4 to 8, and 8 to 12 weeks respectively. 3. These babies acquired their Sarcocystis antibodies which decreased in the first 3 months of life from their mothers. 4. Litters born to serologic Sarcocystis positive mother rats also demonstrated Sarcocystis antibodies. The titres of these antibodies were at the same level as their mothers' at birth but reduced gradually so that most of these young rats were negative at the age of 3 months. 5. Suckling rats born to Sarcocystis negative mothers but positive fathers remained negative. 6. Serologic positive mother and father rats still demonstrated Sarcocystis antibodies in the sera at a time when their litters have become negative. 7. Sarcocystis antibodies could be passed onto the newborn from their positive mothers in both man and rats. 8. These antibodies were probably passed onto the newborn through the placenta but their passage through the colostrum and the mother's milk cannot be ruled out.


[Light microscopic studies on the development of Theileria annulata (Dschunkowsky and Luhs, 1904) in Hyalomma anatolicum excavatum (Koch, 1844). II. The development in haemolymph and salivary glands (author's transl)]. Fully differentiated kinetes, average length 17.6 micrometer, appeared in the haemolymph of engorged nymphs usually 17 to 20 days after repletion. Kinetes were observed at first in the salivary glands on day 18 after repletion. The kinetes then transformed into fission bodies of about 10 micrometer in diameter, mainly in type III alveoli and less frequently in type II alveoli. The fission bodies grew up to a size of about 20 micrometer after several divisions of their nucleus. At this time the ticks moulted and no further development occurred until activation. Shortly before infestation the salivary glands began to proliferate, and rapid growth of the fission bodies was observed, especially in young ticks where development of 'infective particles' ('sporozoites') was concluded within two days. Development in feeding adult ticks apparently occurred in four major steps: (1) Division of primary fission bodies (sporonts) into numerous secondary fission bodies ('primary sporoblasts'), (2) division of secondary fission bodies into tertiary fission bodies ('secondary sporoblasts'), (3) production of particles ('sporozoites') by tertiary fission bodies and release of particles into the saliva, and (4) degeneration of fission bodies and their host cell but further release of particles. The host cell was stimulated to giant growth, thus its diameter increased, on average, from 15 to 110 micrometer. Heavy infections resulting from parasitaemias of greater than 40% caused disease and mortality in the tick population. Development was much retarded by aging. In ticks starved for six months 'sporozoites' did not develop before day five to seven of infestation. 'Sporozoites' did not develop before day five to seven of infestation. 'Sporozoites' may not develop at all in six to nine month old female ticks during the infestation period. The significance of the described developmental stages of T. annulata was discussed and a sexual generation postualted. The hypothetic development of T. annulata in its tick vector was illustrated.


The role of certain oxidative enzymes, catalase, and beta-glucosidase on virulence of Cephalosporium maydis. Isolates of Cephalosporium maydis varied in their pathogenicity to D.C. 67 maize cultivar from highly to weakly pathogenic. Highly pathogenic isolates showed lower activity of polyphenol oxidase, peroxidase, cytochrome oxidase, and beta-glucosidase enzymes and higher activity of catalase and dehydrogenase than weakly pathogenic isolates. Enzymes production by the tested isolates increased as the culture age increased; except in case of catalase enzyme, the reverse action was detected. The role of these enzymes in the virulence of C. maydis is suggested and discussed.


[Behavioral reactions in simians after ablation of the pole of the frontal cortex]. In 4 baboons and 3 macaques ablation of the frontal pole (area 10) as compared to other areas of the frontal cortex did not impair previously elaborated instrumental reflexes of different complexity and did not interfere with the elaboration of new ones. An increase in stability, speed, accuracy and intensity of conditioned reflexes was noted. The ablation did not result in motor or vegetative disturbances. In the general behaviour inhibitory elements disappeared along with an enhancement in motor goal-directed activity. It is suggested that frontal poles do not participate in closing-function but control conditioned reflexes and complex behaviour by means of inhibitory influences.


[Allergization of the body by Neisseria meningitidis. I. The problem of the occurrence of N. meningitidis in cases of bronchial asthma of bacterial etiology (author's transl)]. The object of the authors work was the examination of nasopharyngeal swabs on the prevalence of Neisseria meningitidis. A group of 222 patients suffering from asthma bronchiale of bacteriological etiology was under observation in order to be able to prepare an autovaccine. Nasopharyngeal swabs were inoculated on special nutrient media. Neisseria meningitidis was found in 22%, mainly from nasopharyngeal swabs, together with Haemophilus sp. and Streptococcus pneumoniae. In 59% of examined cases the positive findings lasted longer than 1 year. 46% of isolated strains belonged to group B, about 30% of strains were untypable. The problem of allergisation of people suffering from asthma bronchiale with these strains and the question of preparing an autovaccine from Neisseria meningitidis is discussed.


[Fluorescent antibody technique in identification of Neisseria gonorrhoeae-microcolonies grown on membrane filters (author's transl)]. Filter membranes (Nucleopore, No. N060, CPR 01300, pore size 0.6 mu) are inoculated with a pure suspension of N. gonorrhoeae or a Urethral smear from patients suffering from gonococcal urethritis. The membranes are placed on chocolate agar and incubated for three hours. After fixation in formaldehyd, grown microcolonies of N. gonorrhoea are detected microscopically after incubation of the membranes with a fluorescein-isothiocyanate labeled gonococcal antiserum (Difco, charge no. 605907) (Fig. 3--5). The high specificity of the antiserum was shown by lack of reaction with various other neisseriae (except for N. meningitidis, which showed a weak positive reaction) or other bacteria, respectively (table 1). To investigate the reliability of the new method urethral smears of 23 patients suffering from acute or chronic gonorrhoea were examined by culturing for gonococci in the conventional way and by means of the new technique. In all cases gonocococci were detected with both methods. This suggests that the fluorescent antibody technique has at least the same sensibility as the conventional one, and has the advantage that the result is obtained within one day.


[Antagonistic and bacteriocinogenic activity of Meningococci]. Antagonistic and bacteriocinogenic activity was studied in 169 strains of meningococci of various serological groups and with different localization in the human organism at the time of isolation. Bacteriocinogenic activity was revealed in all the meningococcus strains (by delayed antagonism method), and, in addition, antagonistic activity was found in 100 strains. The inhibitory activity was the greatest in meningococci isolated from the cerebrospinal fluid. The data obtained suggest an important inhibitory activity of meningococci, along with their resistance to the antagonistic activity of the nasopharyngeal microorganisms, in the manifestation of pathogenic properties in them.


[Jet method of administration of live brucella vaccine]. A good survival of brucellae in the organism with the development of a stable and prolonged immunity was noted in vaccination of guinea pigs with live brucella vaccine prepared from the Br. abortus strain 19-BA. Local reactions were weak, and no general reactions were observed. In using the jet method of vaccination with the injectors bpi-4 and bi-2 on a limited group of persons (194) the local and general untoward response was weak, but in immunological respect this method proved to be effective when the vaccine was administered in a dose of 2.5 x 10(8) of microbial cells. Jet method can be recommended for express vaccination against brucellosis.


[Method of obtaining crystalline choleragen and choleragenoid preparations]. A new method of concentration and purification developed allows to obtain choleragen and choleragenoid preparations, homogeneous in the immunochemical analysis with regard to the culturing of vibrios producing these substances on the available nonsynthetic nutrient medium. Crystalline choleragen and choleragenoid preparations were obtained from highly purified samples. Choleragen possessed enterotoxic and skin premeability factors; choleragenoid shows no biological activity.


[Determination of the subclasses of monoclonal human immunoglobulins G and of the light chain type with the aid of specific antisera]. Sera of 86 patients suffering from G-myeloma were studied for the purpose of determination of subclasses of monoclone IgG. Investigations were carried out by means of antisera to subclasses IgG by the double diffusion method in gel after Ouchterlony. The following distribution of myeloma Ig was revealed: G1--70%, G2--17%, G3--11%,and G4--2%. In typing of the light igG chains by the method of immunoelectrophoresis, using antisera to the light chains of immunoglobulins of the chi and lambda type it was found that IgG1 chi was encountered more frequently than IgG1 lambda (3:1 ratio). The amount of the sera with the IgG2, IgG3, and IgG4 was insufficient for the reliable conclusion of their distribution by the type of light chains.


[Effect of brucellosis protective antigen on the phagocytic activity of the cells]. The authors present the results of study of the protective function of cells of the local inflammatory focus caused by the administration of live brucella culture to guinea pigs immunized with a protective antigen in the optimal (0.6) and increased (2 mg) doses. Macrophages of guinea pigs vaccinated with the optimal immunized dose of the protective antigen phagocytized brucellae actively; an increased dose of the antigen suppressed the ingestive and the digestive functions of macrophages, this apparently being caused by the manifestation of the immunodepressive activity of the residual amount of lipopolysaccharide (endotoxin) in the protective antigen.


[Isolation of antilymphocytic antibodies with the aid of cellular immunosorbents]. The authors suggest a method of obtaining purified antilymphocytic antibodies by mean of a specific immunosorbent of human or mouse lymphocytes fixated with glutharic aldehyde Such immunosorbents subjected to special treatment could be used repeatedly; their sorptive capacity was retained in such case. Only from 5 to 12% of the activity could be obtained from immunosorbents sorbed from the serum with 77--93% activity. In comparison with the initial serum, purification was from 6- to 15-fold. Thus, the suggested method provided considerable purification of the antilymphocytic preparations and permitted to obtain highly active antilymphocytic antibodies.


[Indirect hemagglutination test at the ultrastructural level]. A study on ultra-thin sections was made of the preparations of agglutinate produced during the reaction of the immunoglobulin erythrocytic diagnostic agent with dry corpuscular Rickettsia prowazeki antigen, fluoresceine isothiocyanate labeled, and also SRBC used for the preparation of the diagnostic agent after formalinization, tannin treatment, sensitization with hyperimmune horse serum immunoglobulins and lyophilization, respectively. Formalin and tannin treatment of erythrocytes failed to be reflected on the ultrastructure of their cellular membranes; the treatment with hemosensitin was accompanied by the appearance of spheroid protrusions of the erythrocyte cytoplasmic membrane with the preservation of its three-layer structure. Specific interaction of sensitized erythrocytes with the antigen corpuscles was expressed morphologically in their apposition or connection through a gap of 20--30 nm.


[Epiedmiologic significance of the distribution of corynephages in different collectives]. Corynephage distribution was studied in the nasopharyngeal washings of 252 persons infected with C. diphtheriae of gravis type, toxigenic (21 patients and 147 carriers) and non-toxigenic ones (84 carriers), and in 468 uninfected persons in collective bodies under different epidemic conditions. Corynephages were isolated from the nasopharyngeal washings only in persons infected with toxigenic C. diphtheriae--in 4 (of 21) patients, and in 21% (of 147) carriers. Phages tox+ (4--6.2%) were revealed only in carriers of toxigenic C. diphtheriae with numerous bacteria in the nasopharynx and in diphtheria patients. Carriers of nontoxigenic diphtheria bacilli can become infected with phage tox+ only together with the toxigenic strains (reinfection). The data obtained indicated that toxigenic and nontoxigenic C. diphtheriae strains were individual variants.


Orientation of Euglena gracilis by electromagnetic fields: theory and experiment. Computer data derived from theoretical treatments of the orientation of ellipsoidal particles in alternating fields is compared with experimental data from microscopic studies on Euglena gracilis, and elongated free-living flagellate. Computed data based upon a theoretical treatment by SAITO, SCHWAN and SCHWARZ showed good agreement with experimental results, predicting the relationship between cellular orientation, the frequency of the impressed field, and the conductivity of the suspending medium.


Intracellular proteinase of Bacillus subtilis. An intracellular serine proteinase was isolated from Bacillus subtilis, strain A-50. The molecular weight of the enzyme is 30000 +/- 1000, its amino acid composition is enriched in glutamic acid residues, the isoelectric point is 4.3, the N-terminal sequence Glu-Leu-Pro-Glu-Gly-Ile-Gln-Val-Ile-Lys-Ala-Pro-Glu-Leu-Xxx-Ala-Gln-Gly-Phe-Lys Gly-Ser-Asx-Ile-Lys-Ile-Ala-Val-Leu-Asx. The enzyme is structuraly homologous with secretory subtilisins.


Relationships between intracellular proteolytic activity and protein turnover in Bacillus megaterium. When incubated in a sporulation medium, the sporogenous strains of Bacillus megaterium degrade proteins at a rate of 4-10% X h-1. The maximal rate of protein turnover is reached after 3-4 hrs at the time of development of forespores and then decreases again. The rate of protein turnover in the asporogenous strain decreases steadily under similar conditions from 3-8% X h-1 at the beginning of incubation to 1% X h-1 after 5-6 hrs in the sporulation medium. The rate of degradation of proteins in vitro in protoplast lysates is similar or higher than the rate of protein turnover. The exocellular, as well as periplasmic proteolytic activity, is suppressed by amino acids more severely than the activity in protoplasts. Mutants devoid of the exocellular proteolytic enzyme contain also less proteolytic activity in the periplasm than in the protoplasts, in contrast to the wild strain. However, their rate of protein turnover, as well as the degradation of abnormal proteins is similar to that in the wild strain. This supports a view that the proteolytic system in protoplasts is involved in intracellular protein catabolism. The periplasmic enzyme can be considered as a kind of the exocellular proteinase.


Intracellular protein catabolism. IX. Hydrophobicity of substrate proteins is a molecular basis of selectivity. Double-labeled cytosol proteins from rat liver (3H in short-lived, 14C in long-lived proteins) were fractionated by using siliconized glass-beads, phenylsepharose and octylsepharose. Always the short-lived proteins are more tightly bound to the hydrophobic matrix. The same distribution was found with monkey liver substrate proteins. Therefore it is concluded that the different degrees of exposure of superficial hydrophobic areas on substrate protein molecules are a molecular basis of selectivity of the intracellular protein catabolism.


[Neurinomas of the auditory nerve in children]. Analysis of 7 child cases with neurinoma of the acoustic nerve, tumors which are extremely rare in children, showed that they have characteristic clinical features. The disease was manifested at the age of 12--15 years, sometimes against the background of diffuse neurofibromatosis. The mean duration of the disease varied from 5 months to one year. The tumor was marked by rapid growth, medial variant of localization with extension in the oro-caudal direction, and rich blood supply. Hearing diminished rapidly to total deafness, and the loss was usually bilateral. The facial nerve suffered less frequently. The hypertensive-occlusive syndrome was observed with marked stem-cerebellar symptoms and protein-cellular dissociation in the cerebrospinal fluid, which suggests tumor of the cerebellum. Ventriculography with maiodil emulsion and vertebral angiography helped in making a precise diagnosis. Radical removal of the tumor proved very difficult because it was large and intimately connected with the brain stem.


Different  behaviour of 19S thyroglobulin extracted from normal thyroid, simple goitre and toxic goitre in polyacrylamide slab gel isoelectric focusing. 19S thyroglobulin (19S TG) was isolated from normal thyroids, simple goitres and one case of toxic goitre (Graves' disease). With isoelectric focusing on polyacrylamide gel slabs (pH 4--6) several bands for each specimen were detected. The migration patterns of the toxic goitre 19S TG were clearly different from the patterns of normal 19S TG and simple goitre 19S TG.


The measurement of serum proteins with rapid turnover for early diagnosis of fatal hepatitis. An attempt was made to find reliable indices for the early recognition of fatal cases of acute viral hepatitis, using the values of serum proteins with rapid turnover. Prealbumin and alpha2-HS-glycoprotein were measured in the sera of 44 cases by immunodiffusion method before the appearance of hepatic coma and/or gastrointestinal bleeding. The difference of the mean values of prealbumin between fatal and surviving cases of subacute form of fulminant hepatitis was not statistically significant. In contrast to this, there was statistically significant difference between both groups in the mean values of alpha2-HS-glycoprotein (p less than 0.05). The present results indicate the possibility of differentiating fatal cases from surviving ones at an early stage, using the reduction of alpha2-HS-glycoprotein by a simple and reproducible immunodiffusion method.


Structure and ultrastructure of cultivated glial cells from Corpus callosum. 1. Explants and dissociated cells from Corpus callosum (c. c.) of rats and rabbits were cultivated in Petri dishes and Rose chambers. 2. Different types of glial cells were found in the cultivated Corpus callosum (c. c.) explanted from 12 days old rats: a) adendritic glial cells, typical for migrating oligodendroglial cells, b)-migrating large, nondifferentiated astrocytes with pronounced phagocytosing activity, c) macro- and microglial cells which differentiated during cultivation. 3. The population of differentiated glial cells is mostly composed of oligodendroglia, less of astrocytes and microglial cells are rare. 4. Differentiation of dissociated cells from c. c. in homogenous and mixed population was studied. The appearance of first processes of macroglial cells is postponed to 6 to 10 days of cultivation. No substantial difference was observed between homogenous and mixed population. A higher incidence of macrophages was observed in the later. 5. Glial cells differentiate surrounded by degenerated nerve fibers and myelin, exhibiting phagocytoses and cleaning reaction.


Differentiation of corpus callosum glial cells and factors influencing their maturation in vitro. 1. Explants of Corpus callosum (c. c.) from 12-day-old rats were cultivated under different experimental conditions. 2. Migration and differentiation is activated by the presence of neighbouring explants, toward which glial cells predominantly migrate. Glial cells migrate if closely adhering to the supporting collagen and the process of differentiation is enhanced by presence of underlying cell layers. 3. Migratory activity of glial cells decreases and is delayed with age of donors. Migrating cells have a similar appearance as in cultures from 12 days old donors. The presence of immature types of glial cells in c. c. of adult animals was proved. 4. Glucose was found to be an adequate metabolical substrate, utilisation of glucose being lower than in cultivated neurons. In the absence of glucose or serum in the medium, neither migration nor differentiation of glial cells was observed. 5. The addition of embryonal extract and embryonic brain extract enhanced only initial stages of cell migration and differentiation.


The benzidine technique for histochemical detection of the SO4 ion. The benzidine technique for histochemical detection of the SO4 ion was studied analytically on filter paper strips loaded with 0.02 ml of solutions of sulfate containing and sulfate free substances. The influence of the benzidin solution pH on the SO4 and PO4 ion precipitation was analysed and the experimental condition where the SO4 ion was specifically precipitated was established. In basis on this study a benzidine histochemical technique suitable to detect the SO4 ion without interference of the PO4 ion was proposed for tissue sections. This technique has some advantages on the other benzidin techniques previously proposed mainly concerning its specificity.


Ultrastructural labeling of lymphocyte surface immunoglobulin: the preparation and use of soluble immune complexes as indirect immunoelectromicroscopic markers. 2 distinct macronuclear markers, ferritin and hemocyanin, may be used in a mixed anti-globulin labeling reaction to localize lymphocyte surface immunoglobulin (Ig) determinants by transmission electron microscopy. Soluble immune complexes of the marker molecules (antigen) are prepared by complexing with specific antiserum in 40 to 50 x antigen excess; uncomplexed Ig is removed by ultrascentrifugation and/or gel filtration chromatography. Immunoelectrophoresis, spectrophotometry and passive hemagglutination inhibition assay are used to determine the purity and amounts of antibody-antigen in the purified immune complexes. For immunoelectron microscopic labeling, the immune complex markers are coupled to lymphocyte surface Ig by an indirect anti-Ig or anti-allotype antibody linkage. Labelled Ig determinants at 0 degrees C or in the presence of sodium azide are visualized as small patches of marker molecules on the lymphocyte surface membrane. This EM labeling method results in much more consistent and generally higher percentages of surface Ig positive cells (60 to 70% of rabbit peripheral blood lymphocytes) than the percentages obtained using other methods, such as immunofluorescence or autoradiography. If the lymphocytes are warmed to 37 degrees C in the absence of azide the labeled surface Ig determinants undergo rapid endocytosis; endocytotic vesicles containing marker molecules are visible. This mixed anti-globulin immunoelectronmicroscopic labeling system may be used to localize a wide variety of antigens on different cell surfaces.


Immunochemical specificity of a benzidine technique proposed to catalases histochemical detection. As an attempt to test the specificity of an histochemical technique proposed to detect catalases, an investigation was carried out by immunochemical techniques. Purified catalases were used after analysed immunochemically by double immune diffusion test and immunoelectrophoretic technique. These pure catalases induced, after injecting into guinea-pigs, anti-serums that react specifically with catalase and does not give any cross reaction with peroxidases and haemic iron containing compounds. By the direct and indirect immuno-fluorescence techniques it was shown an intense catalase reactivity inside the cytoplasm of adrenal cortex and hepatic cells, that appears as a granular pattern. These results are very similar to those provided by the histochemical technique, either concerning to the reactive cells or to the granular pattern of the positive reaction. In such instances, the immunochemical results suggest the specificity of the histochemical reaction. This specificity is confirmed by the previous treatment of tissue sections by catalases anti-serum. After this treatment either the immunochemical or the histochemical technique to detect catalases provide negative results on cells that before the treatment were strongly reactive.


Histochemical studies on the morphology of the Golgi apparatus and its relation to catecholamine biosynthesis in the locus coeruleus of Rhesus and crab-eating monkeys. Detailed histochemical studies have been conducted on the distribution of thiamine pyrophosphatase (TPPase), L-gulonolactone oxidase (GLO), and NAD-linked xylitol dehydrogenase (XDH) in every component of the Locus coeruleus (LC) of healthy adult male Rhesus and crab-eating monkeys in order to clearify the morphology of the Golgi apparatus (GA) and its relation to biosynthesis of catechol-amines in this special nucleus of the primate (NA nucleus A 6 as defined in the Rhesus monkey by German and Bowden [1975]). Medium-sized neurons of both species of monkeys, which are considered to play an important role in the LC, were classified into 5 groups on the basis of morphological patterns of the GA. Many neurons of both species of monkeys were positive for the XDH test while some neurons of the crab-eating monkey as well as a few neurons of the Rhesus monkey were positive for the GLO reaction. The LC of both species of monkeys must be composed of metabolically one kind of identical medium-sized parasympathetic neurons whose GA may continously undergo 5 distinct phasic changes depending on the functional state of that cell. However, the GA changes its shape much more significantly even within each group of the 5 in the crab-eating monkey than in the Rhesus monkey. The GA Type IV may correspond to the catabolic phase of the GA during which biosynthesis of both catecholamines and vitamin C should be going on. Production of vitamin C may greatly help biosynthesis of catecholamines in LC. The difference in species is evident between the 2 kinds of monkeys studied in regard to the degree of their ability to synthesize these substances. The degree of the ability to synthesize vitamin C parallels the density distribution of Type IV neurons in LC whose GA often develops much more greatly in the crab-eating monkey than in the Rhesus monkey.


Some histochemical observations on the myocardial metabolism in experimental conditions. Part II. A histochemical evaluation of the activity of chosen oxido-reductive enzymes of the cardiac muscle connected with the citric acid cycle, glycolysis and pentose shunt in the early experimental alloxan diabetes period in white rats has been carried out. Dehydrogenases: succinate, isocitrate and glucose-6-phosphate indicated a decrease in the enzymatic activity while the lactate dehydrogenase activity did not undergo any change. An increase in the intensity of the histoenzymatic reaction only concerned glutamate dehydrogenase. The presented histoenzymatic evidence of changes in the enzymatic activities may speak for the possibility of an existence of a direct diabetes influence on the cardiac muscle metabolism.


Studies of estrogen induced renal tumours in male Syrian hamsters. I. Cytochemical studies of lysosomal enzymes acid phosphatase and beta-glucuronidase. The localization and concentration of lysosomal enzymes acid phosphatase and beta-glucuronidase in kidney cells of Syrian hamsters after DES administration were studied. A positive cytoplasmic reaction was demonstrated for both enzymes. Increased activity of these enzymes were observed in renal tumour in contrast to normal homologous cells. These results can be explained on the basis that lysosomal enzymes were synthesized in tumour cells at a higher rate or it may be due to the invasion of tumour tissue by hydrolase rich microphages.


Coated vesicles in the rat anterior pituitary gland. Ultrastructural documentation of the formation of coated vesicles in prolactin cells of the rat anterior pituitary gland is presented. Coated vesicles were observed forming from the secretory granular membrane in the Golgi area and between the Golgi complex and the plasmalemma. They were also found budding off the membrane lining the exocytotic pocket. Following peroxidase reaction product could be seen forming from the membrane of the exocytotic pocket and adjacent to it. These vesicles were found in transit towards the Golgi area or within lysosomes. It is proposed that these coated vesicles are important for membrane conservation and that this is occurring in developing secretory granules near the Golgi complex, in more mature secretory granules and at the level of the plasmalemma during exocytosis.


[The applicability of the glyoxylic acid technique for the demonstration of biogenic amines on the cultured cerebrocortex of the rat (author's transl)]. The glyoxylic acid technique for the demonstration of biogenic amines according to Lindvall and Björklund (1974) was tried to be adapted to the explant culture of the cerebro-cortex of neonatal rats. The "heat reaction" does render the investigation of the explant and of the growth area as well.


[Characterization of heavy metal secretory granules in the islet tissue of adult white rats by means of the light and electron microscopy, including the high voltage electron miscroscopy and the electron microanalysis (author's transl)]. The metal secretory granules of Langerhans' islets of white rats were examined after histochemical detection by sulfide silver staining method as well as by conventional electron microscopy and high voltage electron microscopy. Zinc and calcium were determined with electron probe x-ray-microanalysis. The metals are located at the inside of cytoplasmatic vacuoles, a result, we discuss in connection with the release of pancreatic hormones.


Circadian oscillations of nucleic acid and protein content in functionally different neuron-neuroglia units of rat central nervous system. Scanning integrating visible cytospectrophotometry of gallocyanin-, amido black-, fast green- or ammoniacal silver-stained sections of various areas of the central nervous system of adult rats sacrificed at 4 h intervals has demonstrated the presence of circadian changes in RNA, total protein and total histone content as well as in arginine-rich to lysine-rich histone ratio in the neurons and in their perineuronal satellite neuroglia. The peak of the content per cell of the macromolecular components studied was in some cases elevated above their lowest level up to 1.7-fold. The zenith in the content of neuronal RNA coincided as a rule with the nadir in the content of glial RNA whereas no such opposition was observed with respect to the protein or histone content as well as to the arginine- to lysine-rich ratio in the neurons and in their glial satellite cells. The data obtained have confirmed earlier observations that under changed conditions of neuron functioning, the changes in the nervous system metabolism occur which are localized not only in the neurons but also in the neuroglia, the metabolic response in the glial cells being characterized, depending on peculiarities of the particular neuron activity, both by similarity and by some differences as compared with the neuronal metabolic response. A synchronizing effect of environmental and/or internal factors on the macromolecular metabolism within various cell populations of the central nervous system is discussed.


Diaminobenzidine oxidation in cerebellar histological sections. Cerebellar histological sections were incubated in media containing 3,3'-diaminobenzidine (DAB). Microscopic observation showed the presence of oxidated DAB in all cerebellar cells. Said oxidation of DAB in the cerebellar structures was produced in the absence, as well as in the presence, of exogenous H2O2 in the incubation medium. The erythrocytes existent in the blood vessels were capable of oxidating more intensely the DAB when H2O2 was present in the incubation medium. When the sections were pretreated and incubated with CNK, sodium azide and perodic acid, the oxidation of DAB was not produced. The observation with electron microscope showed that the DAB oxidation is produced in mitochondria, coated vesicles, and lysosomes. The possible mechanisms intervening in the oxidation of DAB in cerebellar structures are discussed.


[On the histochemistry of xanthine dehydrogenase (author's transl)]. We have investigated the enzymes of the XDH-XOD-system by histochemical technique based on a model study for histochemical tetrazolium reduction by flavine enzymes. We have not found a reaction in native sections from liver, kidney and duodenum of adult male Wistar rats. The effectors PMS, Triton X-100 and Tween 80 were inefficient. A strong reaction was observed after a short fixation (10 min) of the sections with glutaraldehyde predominantly in the duodenal epithel cells. The intensity of the reaction was much stronger in sections of liver, kidney and duodenum by influence of the effector PMS and the detergents Triton X-100 and Tween 80. The application of the membrane technique does not show advantages in the histochemical demonstration of the XDH-XOD-system. The specificity of the reaction is discussed under the aspect of the complexity of the XDH-XOD-system shown by biochemical investigations.


Ultrastructural identification of neural elements containing trace metals. The quantitative distribution of heavy metals in the central nervous system was studied by atomic absorption spectrophotometry. It was shown that, of the regions studied, the archicerebellar cortex contains the highest levels of metals. A light microscopical histochemical technique (TIMM silver sulphide method) reveals the neurons and glial cells which possess the trace elements. Ultrastructurally, the staining occurs in different axon terminals, lysosomes, neuronal surface membranes and the cytoplasm of glial cells.


Circadian rhythms of oxidoreductases in the rat gastric mucosa. Histochemical study. Examination of selected oxidoreductases (succinate dehydrogenase, mitochondrial glycerol-3-phosphate dehydrogenase, lactate dehydrogenase, NADH tetrazolium reductase) in the rat gastric mucosa revealed diurnal fluctation of enzyme activities with the most marked manifestation in succinate dehydrogenase. The maximum enzymatic activity found at 18.00 h and 24.00 h) points to the highest oxidoreductase capacity in the parietal cells just at the time when a rat usually expresses spontaneously the highest interest in food intake. The high activity of succinate dehydrogenase and other enzymes at that time is very likely the expression of a "fixed" metabolic adaptation of the parietal cells to the elevated production of hydrochloric acid, in connection with its role in the digestion of food in the stomach. The low enzymatic activity of most rat parietal cells during the day may represent the picture of "a resting afunctional".


[A improved demonstration of ischemic myocard injuries with a combined method for evidence of the succine dehydrogenase and the acridin-fluorchrome-technique (MS-AO-technique) (author's transl)]. A method is described for combined enzyme histochemical and fluorescence microscopical investigation of ischemic injuries. This new method allows to distinguish between 1 reversible and 3 irreversible zones at the experimental ligature infarct of 48 h duration. Heterolysis is discussed as reason for the 3 outer zones and autolytic processes for the central area.


Electronmicroscopic cytochemical studies on lysosomes and peroxisomes of a rat cardiac muscle in the experimental alloxan diabetes. The combined electronmicroscopic and cytochemical investigations of lysosomes and peroxisomes of a rat cardiac muscle were carried out in various duration time of the alloxan diabetes. The analysis of cytochemical reactions with DAB, confirming the existence of peroxisomes in rat heart muscle revealed that the state of experimental alloxan diabetes did not at all affect either cytochemical or morphological image of peroxisomes. In comparison with the control group, in all 3 experimental groups cytochemical reaction to acid phosphatase markedly pointed to the increase in number and size of electron-dense bodies (lysosomes) with positive reaction. This state may most probably result both from local and general organism disturbances in metabolism, induced by particular experimental conditions.


Studies on the oxidizing system in Holt's medium for histochemical demonstration of esterase activity. Esterase activity in guinea-pig thyroid and mouse epididymis epithelial cells has been studied using 5-bromoindoxyl acetate as substrate. The pattern of esterase activity in the thyroid of the guinea-pig is constant, irrespective of whether ferri-ferrocyanide (FFC) or certain copper compounds are used as oxidizing agents in the incubation medium. The intensity of the coloured reaction product is increased when cobalt or manganese are added to the incubation medium. Activity is depressed by high concentrations of FFC when resent in incubation medium or preincubational buffer only. Epididymis cells contain an esterase activity which is not inhibited by conventional SH blocking agents, nor by high concentrations of FFC. From these results it appears that the mode of action of FFC in Holt's medium is as follows. At low concentrations FFC appears to act primarily as a catalytic agent in oxidation of indoxyl to indigoid. At high concentration FFC acts as an inhibitor of guinea-pig thyroid esterase, by oxidation of SH groups in the active centre. The esterase of mouse epididymis cell type EH 1 is not subject to this inhibition by FFC, presumably because it does not contain accessible SH groups in the active centre.


[About the differentiation of antibodies against the connective tissue (author's transl)]. Unfixed cryostat sections of an organ block comprising the liver, stomach, and kidney of rats as well as the kidney of mice were used as antigenic substrate for detecting antibodies against connective tissues by indirect immunofluorescence. There were found more than 14 different fluorescent patterns. Two types could be distinguished easily: The "type 1" shows a clear fluorescence of fine perivascular fibres and a faint fluorescence of vascular endothelium in the liver and in the kidney. The sinusoids are negative. The "type 2" is characterized by a bright and broad fluorescence of the sinusoids and of the portal tract without perivascular staining of the liver. On the sections of the kidney a fluorescence of vascular endothelium, of the tubular basement membrane or of the peritubular connective tissue fibrils is typical.


[A new method for wound age estimation. Histoelectrofocussing (author's transl)]. Histoelectrofocussing is a combination of the usual histochemical methods of wound age estimation with biochemical procedure. Histological sections from skin wounds were directly subjected to isoelectric focussing. The enzymes being extracted and separated were detected by means of histochemical approach. The alterations of the activity of different enzyme fractions can be calculated quantitatively by densitometry. It is possible to differentiate between enzymes of the common skin and those with a topographic connection to the wound edge.


[Comparative study on the dependence of quantitative results in the electron microscopic autoradiography on the type of developer (Microdol X, Phenidon) (author's transl)]. It was the aim of the investigation to compare quantitative results of electron microscopic autoradiographs after the use of different developers (Microdol X, Phenidon) adapting morphometrical methods. Erythroblasts of rabbit were tested after incorporation of 3H-uridine in vitro. Both developers showed the same tendency at higher rates of incorporation (basophilic erythroblasts and polychromatic erythroblasts). Using Phenidon, however, we could obtain only half the values for the individual cell components. At high rates of incorporation Microdol X results in an overlapping of the silver filaments. Thus, more distinct grains can be reached by Phenidon. At smaller rates of incorporation (orthochromatic erythroblasts) after using Phenidon a quantitative evaluation is however influenced by too many factors of uncertainty so that according to our investigations Microdol X is better suited for a quantitative evaluation of electron microscopic autoradiographs.


[The correlation of in vivo and in vitro effect of cytostatics in transplantated tumours of the mouse (author's transl)]. In 2 inoculated mouse tumours the conformity of the actions of 4 different cytostatics at 2 dose levels was tested in vitro and in vivo. The histochemical reaction of lactate dehydrogenase and NADH diaphorase was taken as a criterion of the cytostatic action. The investigations showed the tumours used responded to the cytostatics employed. A good conformity of the in vitro and in vivo findings was established. The histochemical assessment of organ cultures following the administration of cytostatics allows to determine the sensitivity of tumours and helps the physician in deciding on an effective therapeutic.


Microfibrils in the myotendon junctions. Myotendon junctions in the rectus abdominis muscles of bull frogs were examined by the fixation combination of tannic acid and glutaraldehyde using electron microscopy. The features observed on myotendon junctions were the following: (1) There were many deep invaginations of muscle cell membrane at the end of the muscle fibers. Terminal thin filaments of myofibrils were attached to the electron-dense layer lining under the muscle cell membrane on the lateral walls of invaginations. (2) The basement membrane covering the muscle cell membrane was thicker in the invaginations than on the other sites of muscle fibers. (3) Collagen fibers in the invaginations gradually tapered off toward the bottom of the invaginations. But it was not seen that the collagen fibers were attached to both the basement membrane and cell membrane of muscle cells. (4) On the observations using the tannic acid-glutaraldehyde fixation, it was clearly seen that the microfibrils extend from the outer leaflets of the cell membrane to the collagen fibers in invaginations via the basement membrane. It was concluded that the myofibrils might be fastened to the collagen fibers of the tendon by the intermediates of the microfibrils.


Wernicke's encephalopathy in infancy. Development during parenteral nutrition. Wernicke's encephalopathy documented by characteristic involvement of the mammillary bodies developed in a premature infant during parenteral nutrition. This case illustrates the need for thiamine supplementation in seriously ill patients with inadequate intake of the vitamin.


Cross-reactions between Neisseria meningitidis and twenty-seven other bacterial species. Cross-reactions between antigens from N. meningitidis and antigens from 27 other bacterial species from clinical isolates representing 20 different genera were studied by quantitative immunoelectrophoretic methods. A water-soluble N. meningitidis antigen preparation and a corresponding pooled rabbit antiserum regularly presenting 48 immunoprecipitates were used as reference system. Antigens from 16 other bacterial genera cross-reacted with one to four N. meningitidis antigens, whereas antigens from six other Neisseria species cross-reacted with 37 to 48 N. meningitidis antigens. Antigens from only three bacterial genera (Streptococcus, Staphylococcus and Bacteroides) did not show any cross-reactions. The degree of cross-reactivity between the antigens were 25--100%, as judged by absorption of antibodies. Three of the cross-reacting antigens reacted with normally occurring precipitating antibodies in serum from humans and rabbits.


Antigenic characterisitcs of Moraxella nonliquefaciens fimbriae in double immunodiffusion. Rabbit antisera against purified fimbriae (pili) from Moraxella nonliquefaciens detected three fimbrial antigenic components, one or two of which appeared to be present in other fimbriated strains of M. nonliquefaciens and the closely related M. bovis. Maximal precipitation with the antisera required some denaturation of the antigen. Ultrasonication, repeated freeze-thawing, heating, and agents like KBr, NaSCN and urea were effective in liberating the antigen in diffusible forms. The morphology of the fimbriae was altered by heat treatment in 1 M KBr.


Assessment of wound contamination by wound irrigation. Experimental investigations on quantitative recovery of anaerobic and aerobic bacteria. The quantitative recovery of E. coli, S. faecalis and B. fragilis from operative abdominal wounds was investigated in pigs in an experimental model suitable for statistical calculations. Wounds were contaminated in groups of ten with different numbers of either a single bacterial species or a mixture of two species. The wound was irrigated with saline 20 minutes after contamination. Significant differences in recovery were found between the bacterial species investigated. Expressed as percentage of the number of bacteria used for contamination, the recovery for a given species was rather low, but it was constant and independent of the degree of contamination. The investigation did not suggest any principle difference in the recovery of anaerobic and aerobic bacteria. The clinical applicability of the method is not yet clarified.