Effect of additions of the carbon and nitrogen sources on the NADP+ and NAD+ level in the mycelium of a producing strain of P. nigricans Thom.]

The levels of NADP+ and NAD+ in the mycelium of a cultured strain of P. nigricans Thom grown on the mineral medium with 2 per cent of glucose were studied as dependent on the time and method of addition of glucose. NaNO3 or their mixture to the medium, i. e. a single addition simultaneously with the spore inoculation or on the 7th day of the culture development, divided additions on the 5th--7th days. It was shown that addition of the above sources simultaneously with inoculation affected the levels of both dinucleotides by the 5th and 8th days. Analogous additions on the 7th day in 24 hours decreased the level of NAD+, while the NADP+ concentration increased after addition of a mixture of glucose and NANO3 and remained unchanged on their separate addition. When the above sources were added in divided doses the level of NADP+ increased till the 7th day and that of NAD+ till the end of the experiment (the 8th day).     

The influence of sevoflurane on the reactivity of astrocytes in the course of the experimental intracerebral haemorrhage in rat.

40 adult Wistar rats were divided into two groups depending on the applied anaesthesia. In both groups animals were generally anaesthetized with fentanyl, dehydrobenzperidol administered intraperitoneally and midazolam given intramuscularly. In the second group (SEVO) animals received sevoflurane of 2.2 vol% end-tidal concentration. Intracerebral haematoma was produced through infusion of 100 microl of autologous blood into the striatum. Each group was divided into five subgroups depending on the length of survival period: 1, 3, 7, 14, 21 days. The astrocytic population was studied by means of anti-GFAP staining. Stereological analysis was applied to estimate the numerical density of immunoreactive cells and the distribution of their types. On 7th day of observation the density of GFAP-immunoreactive astrocytes in SEVO was lower (p<0,05) than that in the control group. In the control group, the increase (p<0.05) of per cent of activated astrocytes between the 1st and 3rd survival day was noted, which remained at this level till the end of observation. In SEVO group, the increase (p<0.05) of per cent of activated astrocytes between the 3rd and 7th day and the decrease (p<0.05) between the 14th and 21st survival day were observed. During days of observation the per cent of activated astrocytes was lower (p<0.05) in the SEVO group than that in the control group. Administration of sevoflurane during anaesthesia to animals with intracerebral haemorrhage has evoked not only the delay of the activation of astrocytes but also decrease in its level.     

Morphogenesis and pathogenesis of chronic lung diseases. vii. Kinetics of alkaline phosphatase active cells during the development of experimental lung granulomas.

The circulating cells arriving at the formation site of the experimentally induced lung granuloma were quantitatively and qualitatively determined during their development by estimation of alkaline phosphatase active free cells. These represent 32 per cent the 2nd day and 27 per cent the 4th, of all the lung granuloma cells, and rapidly decrease to 8 per cent the 8th day, to 4 per cent the 11th and to only 1.5 per cent the 15th day. The participation of alkaline phosphatase circulating cells appears to be important only the first days, the process becoming by proliferation and differentiation a histiocytic, epithelioidic, and lympho-plasmocytic structure under the influence of antigenic and non-antigenic components of the used immunologic adjuvant.     

Mortality rates in patients with burns; a report of experience at San Francisco City and County Hospital, 1943-1956

A review was made of the cases of 93 patients with burns covering more than 20 per cent of the body surface who were treated at the San Francisco City and County Hospital, University of California Service, between 1943 and 1956. The mortality rate increased from 40 per cent during 1943-1947 to 69 per cent during 1952-1956. A significant change in survival time was noted: During 1943-1947, 69 per cent of the deaths occurred within 48 hours of admission; during 1952-1956, only 19 per cent of the deaths occurred within the first 48 hours. In the period 1943-1947 the majority of deaths resulted from shock in the immediate post-burn period; in the later years of the study the major cause of death was infection. No patient more than 50 years of age who had burns of more than 25 per cent of the body surface survived. Only one patient with burns involving more than 45 per cent survived. No patient who had a blood culture positive for bacteria survived. The use of antibiotics had no effect on the incidence of infection. Elderly patients, children and alcoholics were less able to resist the effects of infection. The lowest mortality rate was in the age group of 15 through 35 years.     

The viability and survival of sporozoites of Eimeria in vitro

Some factors affecting excystation and viability of sporozoites of several species of Eimeria from chickens were examined in vitro. Chicken embryos or cultured kidney cells were inoculated with sporozoites in order to assess viability.Sporozoites of E. tenella survived in phosphate buffer (P.B.S.) containing 0·9 per cent NaCl for 14 days. Some sporozoites survived in solutions containing up to 16 per cent NaCl for 3 days at +4°C. Sporozoites of E. maxima and E. acervulina survived for only 27 h in phosphate buffer containing 1 or 2 per cent NaCl.Sporozoites of E. brunetti, E. maxima, and E. acervulina var: mivati were released rapidly from sporocysts in vitro, but survived for relatively short periods in PBS at 4°C. However, the addition of serum or gelatine to these solutions increased survival to at least 96 h.The viability of sporozoites after freezing and storing in liquid nitrogen was best when 12 per cent dimethyl sulphoxide (DMSO) was added to the sporozoite suspensions. P.B.S. with DMSO was less suitable than the other solutions used and serum or gelatine with the DMSO, was needed to increase survival. Increasing the density of sporozoites in the frozen stabilates did not increase survival.     

Glycogen metabolic enzymes in the skeletal muscles of the developing chick embryo

In the skeletal muscles of the chick embryo from the 10th till the 15th day of embryogenesis, phosphorylase (EC. 2.4.1.1) is represented by two isozymes one of which corresponds, by electrophoretic mobility, to the liver phosphorylase and another to phosphorylase of the skeletal muscles of the adult rat. From the 17th day of embryogenesis on only one isozyme of phosphorylase is found in the skeletal muscles which is identical with that of the skeletal muscles of the adult bird. The isozyme spectrum of phosphorylase of the whole 4 days old embryo contains, besides phosphorylase L, a special "embryonic" isozyme which differs from that of the skeletal muscles by immunochemical characteristics and electrophoretic mobility. From the 10th day of embryogenesis till hatching, the activity of phosphorylase of the skeletal muscles increases more than 50 times and that of glycogen synthetase (EC. 2.4.1.11) only 4 times.     

Pharmacokinetics of dioxidine in oncological patients in the postoperative period

Dioxidine pharmacokinetics was studied in 5 patients operated for cancer of the large intestine and treated prophylactically with the drug during the postoperative period. Dioxidine was administered intravenously for 10 minutes twice a day in an amount of 300 mg in 5 per cent solution of glucose. The drug concentrations in serum and urine were determined with a microbiological procedure. Escherichia coli AB 2472 rec A16, a strain deficient with respect to reparation was used as a test microbe. The plates with the dilutions were incubated under anaerobic conditions. The time course of the drug concentrations in serum was shown to be satisfactorily described by the following equation: C(t) = 3.125 . 1-2.57.t + 2.76 . 1-0.64.t. Within the first 1.5-2 hours after the administration the dioxidine concentrations in serum and urine amounted to 2.5-4 and 35-50 micrograms/ml respectively.     

Evaluation of the co-agglutination test in diagnosis of experimental microsporidiosis

Microsporidiosis is an emerging and opportunistic infection associated with wide range of clinical syndromes in humans. Confirmation of the presence of microsporidia in different samples is laborious, costly and often difficult. The present study was designed to evaluate the utility of the Co-agglutination test (Co-A test) for detection of urinary, fecal and circulating microsporidial antigens in experimentally infected mice. One hundred and twenty male Swiss albino mice were divided into non infected control and infected experimental groups which were further subdivided into two equal subgroups; immunosuppressed and immunocompetent. Microsporidial spores were isolated from human stools and identified to be Encephalitozoon intestinalis by the molecular methods. They were used to infect each subgroup of mice, then their urine, stools and sera were collected at the 1st, 3rd, 5th, 7th and 9th days post-infection (PI). Co-A test, using prepared hyperimmune serum, was used to detect antigens in all samples collected. The cross reactivity of microsporidial hyperimmune sera with antigens of Cyclospora cyatenensis and Cryptosporidium parvum was investigated by Co-A test. The results showed that Co-A test was effective in detecting microsporidial antigen in stool of immunosuppressed infected mice from the 1st day PI, and in urine and serum from the 3rd day PI till the end of the study. In the immunocompetent subgroup, Co-A test detected microsporidial antigens in stool, serum and urine of mice from the 1st day, 3rd day and the 5th day PI, respectively till the end of the study, without cross reactivity with C. cyatenensis or C. parvum in both subgroups. Co-A test proved to be simple and suitable tool for detecting microsporidial antigen in different specimens and did not need sophisticated equipment. It is very practical under field or rural conditions and in poorly equipped clinical laboratories.     

Leukemia L 1210 cells induce depletion of Lyt 2+ thymocytes

The thymocytes were analyzed on the 7th day after i.p. inoculation of 10(6) leukemia L 1210 cells to syngeneic DBA/2 Wf mice. A three-fold decrease of the total number of thymocytes was found as well as 1.7-fold decrease of the per cent of thymocytes with Lyt 2+ phenotype, while the per cent of cells with phenotypes Thy 1.2+ and Lyt 1+ was unchanged.     

CHANGES IN THE CONTENT OF GIBBERELLIN-LIKE SUBSTANCES IN RIPENING SEED AND POD OF LUPINUS LUTEUS

The amount of gibberellin-like substances in seed, pod, embryoand "endosperm" of Lupinus luteus in relation to their developmentwas studied by means of the rice seedling test. The amount of gibberellin-like substances per seed increasedremarkably at the first stage of development, when the growthof seed and pod was very slow, and attained its maximum at suchan early stage as the dry weight of seed reached a value ofonly 10 per cent and that of pod 28 per cent of their respectivemaximum levels. Changes in amount of gibberellin-like substancespresent in the pod were very closely related to the changesin growth of pod. Gibberellin-like substances equivalent to0.3 µg gibberellin A₃ per seed and 0.028 µg perpod were found at the 20th day after anthesis, and no activityin mature seed and 0.09 µg per pod at the 50th day. Noactivity was found in the embryo itself at all stages, indicatingthat the gibberellin-like substances in seed occur only in the"endosperm," i.e. in the tissues that will eventually form testain fully ripened stage. (Received December 18, 1962; )     

Use of spiramycin in the treatment of inflammatory diseases ot the respiratory tract in children in ambulatory conditions]

Comparative efficacy of oral spiramycin and ampicillin was estimated in the treatment of 65 children at the age of 5 to 12 years with infectious inflammatory diseases of the respiratory tract, tonsils and middle ear. By the 7th day of the treatment with spiramycin the cure was stated in 97.7 per cent of the patients and 2.3 per cent of the patients showed the improvement. With the use of ampicillin the cure was recorded only by the 12th day. Marked advantages of spiramycin were observed as well with respect to the time course of the improvement of the disease main signs such as fever, pain in the throat on swallowing, intoxication and others.     

Ultimobranchial body and parathyroid gland of the parrot Psittacula psittacula in response to experimental hypercalcemia

Psittacula psittacula when subjected to long term hypercalcemia by intramuscular injections of vitamin D2 (20,000 I.U.) on alternate days and by increasing dietary calcium, exhibit a rise in the serum calcium level after 10, 20 and 30 days of treatment as compared to their corresponding controls. The ultimobranchial cells show progressive hypertrophy up to 20th day of the treatment. From 20th day till the end of the experiment (30 days) these cells show feeble staining response. The parathyroid glands suffer from degenerative changes due to its inactivity under chronic hypercalcemia.     

Comparative study of the cytotoxic effects of anthracycline antibiotics on heterotransplants of human breast cancer cultivated in diffusion chambers in vivo

Cytotoxic activity of doxorubicin, daunomycin, carminomycin and ruboxyl against 50 human breast cancer heterotransplants in diffusion chambers was studied. The effect was estimated autoradiographically on the 6th or the 7th day of the cultivation after the drug administration in the maximum tolerance doses. The tumors were considered sensitive when the labeling index of their transplants after the treatment appeared to be reduced by 50 or less per cent against the control. The number of the tumors sensitive to all the drugs amounted to 72-80 per cent. 19 tumors were sensitive to 4 antibiotics. 14 and 8 tumors were sensitive to 3 and 2 antibiotics, respectively, and only 1 tumor was sensitive to 1 drug. The sensitivity significantly correlated with the initial labeling index of the primary tumors and their heterotransplants. The results suggested that daunomycin and ruboxyl possessed a high cytotoxic activity close to that of doxorubicin and carminomycin and might be recommended for clinical trials in the treatment of patients with breast cancer.     

Study of the effect of antibodies in the intestinal tract of germ-free baby pigs

The effect of antibodies in the intestinal tract was studied in germ-free baby pigs whose intestinal barrier was closed to macromolecules by the peroral administration of modified cow's milk for the first 72 hours after birth. They were then all contaminated with the pathogenic strainEscherichia coli 055 in amounts of 10⁹ bacterial cells per animal. The controls, which were not given any antibodies, all died within 24 hours. All the experimental animals given 12.5–50ml immune colostrum or serum survived, while of those given 50ml normal serum or colostrum containing natural antibodies reacting with theEscherichia coli test strain, 50% survived. No circulating antibodies were found in the serum of the experimental animals after the administration of serum or colostrum. The antibodies present in colostrum thus appear to protect the newborn organism directly in the intestinal tract, which is the first site of bacterial invasion, as well as after infiltration into the blood stream.     

A single serum dilution method for the quantitation of neutralizing antibodies to varicella-zoster virus

A one-point serum dilution method for determination of neutralizing antibody in human sera to varicella-zoster (V-Z) virus instead of the serial serum dilution method was investigated. Focus counting was performed under a microscope on day 5 to 6 after inoculation of V-Z virus into 6-well plastic trays in which human embryonic lung cells were grown. A table was constructed to estimate the ND50 titers by the per cent reduction of the focus count from the control at only one dilution of test sera. The estimated ND50 values agreed well with those determined by the serial serum dilution method. Test sera showed a slight nonspecific reactivity at low serum dilutions, but reliable results could usually be obtained at a serum dilution of 1:8 or more. This method, which saves materials and labor, was applied to the quantification of neutralizing antibody against V-Z virus in human sera with satisfactory accuracy and reproducibility.     

Viremia and serum antibodies in Syrian hamster after inapparent infection with European tick-borne encephalitis virus

1.) Adult Syrian hamsters (Cricetus auratus) are relatively insusceptible to the lethal affect of the ETBE virus after peripheral inoculation. After subcutaneous infection with 10(3) i. cer. mouse LD50 of the virus they manifest 25 per cent mortality. 2) In subcutaneously infected hamsters the virus is demonstrable in the CNS of only 20 per cent of the animals. CF- and VN-antibodies can be found in the blood from the 10th day; CF--antibodies culminate on the 20th-30th day receding relatively quickly, whereas VN-antibodies peak after the 28th day, dropping only partly and persisting. 3) The dynamics of CF- and VN-antibodies and a high number of clinically inapparent hamster infections resemble the situation in humans infected with ETBE under natural conditions.     

Demonstration of growth in porcine thyroid cell culture

Eagle's minimum essential medium supplemented with 20 per cent newborn calf serum (N.C.S.) allows porcine thyroid cell survival but not cell growth in vitro. In NCTC 109 medium supplemented with 20 per cent N.C.S. these cells actively grow and may be serially propagated. Cell population doubling time expressed as DNA doubling value is 3.5 days at 37 degrees C in 95 per cent air-5 per cent CO2. Thyrotropin does not affect porcine thyroid cell multiplication in vitro but stimulates the plating efficiency in primary cultures to about 130 per cent of controls. Cell selection was obtained by replacing media with Earle's balanced salt solution. This operation provoked death of nearly all cells by day 18 but subsequent addition of growth medium resulted in proliferation of epithelial cell clones. From generation 2 to generation 8, cells produce thyroglobulin but they do not actively trap iodide nor form follicles when thyrotropin is added to the media. Cell selection, demonstration of growth, as well as freeze-storage techniques described in this paper permit selection and storage of porcine thyroid cells and the potential constitution of cell collections.     

REVIVAL OF MAMMALIAN SPERM AFTER IMMERSION IN LIQUID NITROGEN

1. A wide variety of procedures was used to test the motility of mammalian sperm after plunging them into liquid nitrogen at –195°C. and later rapidly warming them to 35°C. by plunging them into a suitable balanced and isotonic medium. 2. Using seminal fluid sperm from the same human donor, maximal numbers of motile sperm survived vitrification when the samples were (a) very fresh, (b) untreated with plasmolysing solutions, (c) plunged into the refrigerant in the form of a foam. The maximum yield of motile human sperm recoverable from the liquid nitrogen was 50 per cent. Since in this sample only 75 per cent of the sperm were alive before immersion, 67 per cent of the living sperm survived vitrification. 3. Experiments with sperm from 31 rabbits were made with a variety of conditions of pretreatment to obtain maximal yields of recoverable, motile sperm after vitrification by liquid nitrogen. (a) A consistent recoverable yield of about 0.5 per cent was obtained when the untreated suspension of sperm was smeared on cellophane and partially dried in air before immersing in liquid nitrogen. (b) On a few out of many occasions plasmolysis for several minutes with hypertonic Ringer solution gave a recoverable yield of 0.1 per cent as did (c) pretreatment with hypertonic Ringer and butyric acid.     

Early postnatal developmental changes of the cardiac electric field in guinea-pigs.

Changes of the "representative" maximum spatial cardiac vector were followed daily from the 1st till the 3th postnatal day of life in a group of 70 guinea-pigs. This vector representing the spatial vectorcardiographic loop shifted gradually during the observation period from the right caudal quadrant to the left caudal quadrant attaining its new spatial orientation on the 7th day. This shift is accompanied by a transitory tilt in the cranial direction with a peak on the 7th day and a transitory decrease of the spatial magnitude with a peak on the 4th day. There was no significant difference between the spatial magnitude of the "representative" maximum vector on the 1st and the last (13th) day of observation.     

Nerve fiber interrelationships with the ependyma of the 3d ventricle in rats in the perinatal period]

The localization and ultrastructure of nerve fibers in the wall and lumen of ventral region of III ventricle was studied from the 16th day of prenatal period till the 9th day of postnatal period in the Wistar rats. The nerve fibers were first found in the subependyme zone; between the ependyme cells and in the lumen of ventricle on the 18th day of development. The nerve fibers occur constantly in the lumen of ventricle beginning from the 3rd day of postnatal period. The dilatations of nerve fibers contain granules of 50 to 90 nm in diameter with the electron-dense center (carriers of monoamines and/or releasing hormonnes) granules with moderately osmiophilic contents and light vesicles of the same size. It is suggested that these structures reflect different stages of the release of neurohormones in the cerebrospinal fluid. The synapse-like contacts of nerve fibers with the ependyme cells are observed beginning from the 3rd day of postnatal period. Their role in the regulation of absorption of substances by the ependyme cells functioning is discussed.