Two factors affecting the heat stability of xanthine oxidase in extracts of mouse intestine

The heat stability at 56 C of xanthine oxidase (Xox) from mouse intestine was found to be affected by two factors: (1) Xox which had been partially digested by trypsin was less heat stable than Xox which was protected from digestion by the presence of soybean trypsin inhibitor and phenylmethylsulfonyl fluoride; (2) Xox was less heat stable in glass tubes than in polystyrene tubes and siliconized glass tubes resembled polystyrene tubes in this respect.     

Recovery of hematopoiesis in mice following a continuous irradiation with a dose rate of 0.957 Gy/d and a total accumulated dose of 19.14 Gy. I. Bone marrow, spleen and thymus gland

This paper deals with the evaluation of histological changes in the bone marrow, spleen and thymus of mice after continuous irradiation with a dose rate of 0.957 Gy/day and a total accumulated dose of 19.14 Gy. Erythropoiesis in the spleen could be recovered quickly, significantly exceeding the spleen erythropoiesis of the controls on the seventh post-irradiation day. Myelopoiesis in the bone marrow could be recovered until the 21st day and erythropoiesis until the 28th day after the end of irradiation. Lymphopoiesis in the thymus could be recovered on the 28th day approximately and in the spleen roughly on the 60th day after the end of irradiation.     

Recovery of hematopoiesis in mice following a continuous irradiation with a dose rate of 0.957 Gy/d and a total accumulated dose of 19.14 Gy. II. Peripheral blood

This paper deals with quantitative and qualitative changes in the peripheral blood of mice after continuous irradiation with a dose rate of 0.957 Gy/day and with a total accumulated dose of 19.14 Gy. During irradiation a significant diminution of nongranular leukocytes, a granulocytopenia and a decrease in erythrocytes to about 70% of the control values could be observed. Erythrocytes recovered their original state until the 14th day after the end of irradiation, granulocytes until the 21st day and agranulocytes until the 60th post-irradiation day. The leukocyte number decreased significantly again until the 80th day after the end of irradiation. Leukocytes with intense morphological anomalies could be observed in the peripheral blood during the whole period of examination.     

Changes in lipoprotein lipase activity in the adipose tissue and heart of non-lethally X-irradiated rats

After 16 h nocturnal deprivation of food, male Wistar rats were irradiated by a single whole body dose of 2.40 Gy X-rays. Both the irradiated and sham-irradiated (control) rats were pair-fed for the first six days after irradiation, but for the rest of the time they were fed ad libitum. Lipoprotein lipase activity (LPLA) in the adipose tissue fell between 24 and 48 h; LPLA in the heart fell at 24 h and 21 days and rose on the 14th days. The serum triacylglycerol concentration rose between 24 and 72 h. Comparison with the fed control group showed LPLA in adipose tissue to be reduced at 6 and 72 h and on the 28th day and raised between the 7th and the 14th day. In the heart it was raised at 1 h and between 72 h and the 14th day, it was reduced on the 21st day and rose on the 35th day. The triacylglycerol concentration was raised between 48 and 72 h and on the 28th day. Pair-feeding after non-lethal X-irradiation allowed more exact differentiation of the specific effect of ionizing radiation on LPLA in the adipose tissue and heart at the early post-irradiation intervals.     

Use of a modified Robbins device to directly compare the adhesion of Staphylococcus epidermidis RP62A to surfaces

Staphylococcus epidermidis is a frequent cause of infection associated with the use of biomedical devices. Flow cell studies of the interaction between bacteria and surfaces do not generally allow direct comparison of different materials using the same bacterial suspension. The use of a modified Robbins Device (MRD) to compare the adhesion to different surfaces of Staph. epidermidis RP62A grown in continuous culture was investigated. Adhesion to glass was compared with siliconized glass, plasma-conditioned glass, titanium, stainless steel and Teflon. Attachment to siliconized glass was also compared with glass under differing ionic strength, and divalent cation concentrations. Both the differences in numbers adhering and changes in adhesion (slope) through the MRD were compared. There was a trend towards higher numbers adhering to the discs at the in-flow end of the MRD than at the outflow end, probably reflecting depletion of adherent bacteria in the interacting stream. Adhesion of Staph. epidermidis RP62A to siliconized glass and Teflon was reduced when compared to glass with increasing flow rates. Adhesion to stainless steel was not affected by flow rate and titanium gave a different slope of adhesion through the MRD when compared with glass, suggesting an interaction with different sub-populations within the interacting stream. Differences between siliconized glass and glass at flow rates of 300 ml h-1 were abolished by the addition of calcium or EDTA and reduced by the addition of magnesium. Increasing ionic strength reduced the statistical significance of the differences between glass and siliconized glass. Pre-conditioning of glass with pooled human plasma reduced adhesion compared with untreated glass and again gave a different slope to glass. The MRD linked to a chemostat can be used to compare directly bacterial adhesion to potential biomaterials. Variable depletion of the interacting stream should be taken into account in the interpretation of results. Divalent cation concentration, substrate properties and flow rate were important determinants of the comparative adhesion of Staph. epidermidis RP62A to surfaces.     

Lipids and caplain in guinea pig tissues in the process of development of experimental autoimmune encephalomyelitis

The character of some lipids level change--cholesterol and phospholipids--as basic lipid components of cell membranes in the guinea-pig brain and liver tissue, and in serum in conditions of development of experimental autoimmune encephalomyelitis (EAE) have been investigated on the 11th, 21st, 27th day after inoculation. It has been detected, that the level of the investigated lipids changes wavely and indifferent-direction in the brain tissue on the 21st day of EAE. Similar variability observed in the activity of proteolytic ferment calpain, which is authentically reduced in the brain tissue by the 11th hour and increases up to the test objective level in the subsequent periods of EAE development. In the liver the level of alpha-tocopherol is reduced, while the content of studied lipids does not change. The investigated parameters can be attributed to the factors, which play an essential role in structural stability of cell membranes and their variability in conditions of EAE development is related to the processes of nervous cells demyelinisation and, hence to occurrence of such pathology as multiple sclerosis in people.     

Cost and benefit from shield cases in caddis larvae

The seasonal variation of the periphytic community attached to an artificial substratum (glass tubes) was studied during two different periods in a lagoon connected to the Paranapanema River, the main tributary of the Jurumirim Reservoir (São Paulo–Brazil). An analysis of dry weight, ash free dry weight, chlorophyll a, phaeophytin and primary productivity of periphyton was carried out. The first experiment lasted from August to December 1993, the second from February to June 1994. Tubes were removed after 7, 14, 21, 28, 60, 90 and 119 days of incubation. In the 1st experiment, the periphytic community reached a higher biomass after the 4th week of colonization (28th day), in the 2nd experiment after the 2nd week (14th day). This discrepancy is related to seasonal differences in environmental factors (water temperature, nutrients concentrations and water discharge) that determine initial colonization. After the first stages of colonization, the biomass and primary productivity of periphyton reached their maximum values after the 60th day of incubation. In both experiments, three developmental phases could be discerned. In the initial phase, an exponential growth was observed. In the second phase, the bioderm reached its maximum biomass and productivity. In the third phase, a decrease of biomass and productivity occurred.     

Changes in mouse erythropoiesis during continuous irradiation with a daily dose of 957 mGy

Quantitative and qualitative changes in the blood producing organs and in the peripheral blood of mice are evaluated in this paper. The animals were irradiated for 42 days continually with a daily dosage of 957 mGy. Until the 7th day of irradiation a significant diminution of the cellularity of the bone-marrow and of the cellularity as well as the mass of the spleen could be observed. After the 14th day or irradiation a temporary stabilization of the cell number in the bone-marrow could be found until the 28th day, after that time there was a moderately strong decrease. The cellularity and the mass of the spleen increased temporarily until the 28th day of irradiation because of erythropoiesis and myelopoiesis increasing from 20% to 50%. The most significant changes in the peripheral blood could be observed in not granulated cells as a kind of sudden and permanent decrease. The diminution of the granulocyte and reticulocyte number proceeded somewhat more slowly, temporarily revealing an increasing tendency on the 28th day of irradiation. The erythrocyte numbers as well as the haematocrit and haemoglobin values decreased continually beginning from the 7th day of irradiation until the death of the test animals.     

Prolactin inhibition in lactating rats changes leptin transfer through the milk.

Malnutrition during lactation reduces milk production and changes pup's leptin serum levels. To test prolactin role in this nutritional state, we evaluated whether prolactin suppression during lactation changes serum leptin in dams, its transfer through the milk, and pup's serum leptin. Lactating rats were treated with bromocryptine (1 mg/twice a day, s.c.) or saline three days before sacrifice (days 2-4 or days 19-21). Food intake and body weight were measured until sacrifice (4th and 21st day). Serum prolactin and leptin were determined by radioimmunoassay. Bromocryptine injected dams had lower serum prolactin and milk production as expected. The mothers presented lower food ingestion (day 21: -25%), lower body weight (day 4: -12%; day 21: -10%), higher serum leptin (day 4: +68%), lower milk leptin on the 4th day (11 times) and higher (8 times) on the 21st day. The offspring of bromocryptine-treated mothers presented lower body weight in both periods of lactation and lower serum leptin on the 4th day (-40%) and higher on the 21st day (+37%) of lactation. We suggest that prolactin, through its effect on leptin secretion into the milk, may play an important role in signalizing maternal nutritional status to the pups.     

Comparison of the reparative property of early and late CFUs after sublethal radiation injury

Restoration ability of early and late CFUs has been studied using bone marrow sublethal fractional irradiation. It has been shown that CFUs capacity for early postirradiation repair on the 11th day is sharply reduced, as compared to that of CFUs on the 8th day.     

Effects of acute X-irradiation on pre-implantation embryos and on the implantation reaction in the mouse

Summary Pregnant mice were treated on the 1st, 2nd, and 3rd day of pregnancy by a single dose of 300 R X-rays. Uterine dissections at day 6 p.c. topographically revealed decrease of the implantation sites from 9.67 per female in the controls to 8.00 in females irradiated on day 1, to 6.63 in females irradiated on day 2, and to 7.00 in females irradiated on day 3 p.c. Among a number of 22 implantations after irradiation on day 1, 19 after irradiation on day 2 and 11 after irradiation on day 3, however no living embryo could be detected on histological examination. The degree of damage as indicated by the total resorptions was highest (94,7%) after irradiation on day 2 p.c., and lowest (31,8%) after irradiation on day 1 p.c. Since the decidual cell reaction was either unaffected or only slightly reduced after irradiation on day 2 p.c. as indicated by cytomorphological criteria and the alkaline phosphatase reaction, not maternal effects but direct effects only of the irradiation on the embryo must account for embryonic deaths.     

Ultrastructure of cells of rat lymphatic organs during continuous irradiation

Changes in the ultrastructures of lymphatic organs of rats observed from the 2nd to 22nd day following continuous irradiation with gamma rays at a daily dose of 115 mGy (exposition: 12 R/day) are described. The maximum of destructive changes in lymphocytes was observed on the 11th to the 14th day of irradiation. A gradual balance between dystrophic and regenerative processes was achieved on the 18th day. In this connection to correlation could be determined between the ultrastructural changes described and the fluctuations of lymphocyte numbers in the lymphatic organs and peripheral blood during continuous irradiation.     

The influence of prolonged cimetidine administration on serum gastrin levels and gastric acid secretion in rats.

The correlation between serum gastrin levels and gastric acid secretion during 4 weeks of cimetidine administration (once daily) was investigated. Serum gastrin levels and gastric acid secretion were estimated on the 7th, 14th, 21st and 28th day after cimetidine administration (25 mg.kg-1, intragastrically). At the mentioned time intervals gastric acid secretion stimulated by histamine and pentagastrin was also studied. It was found that on the 14th and 21st day after cimetidine administration serum gastrin levels were significantly elevated. Basal gastric acid secretion after cimetidine administration was significantly decreased at all the observed time intervals. Histamine-stimulated gastric acid secretion was increased on the 14th, 21st and 28th day after cimetidine administration. Hypoacidity was not followed at all time intervals by hypergastrinaemia (only on day 14 and 21 after cimetidine).     

Chemical degradation of 3H-labeled substance P in tris buffer solution

Substance P (SP) is an important neuropeptide that has been implicated in several physiological processes, and it is necessary to devise an analytical procedure to measure endogenous SP with a combination of high sensitivity and maximum molecular specificity. However, the unique chemical nature of SP (polarity, chemical stability, ease of oxidation, peptide bond lability) plays a significant role in its analysis, such as in receptor assays, immunoassays, chromatography, and mass spectrometry. In this study, we evaluated in polypropylene and glass assay tubes the effects on the recovery and stability of tritiated SP ([3H]SP) of several pertinent experimental parameters such as buffer, pH, multiple freeze-thaw cycles, and incubation temperature and time. Bovine serum albumin (BSA) effectively reduced the absorption of [3H]SP to polypropylene and glass tube surfaces. Following multiple (6X) freeze-thaw cycles of solutions in BSA-precoated tubes, the recovery of radioactive [3H]SP remained high (greater than 75%) after the last cycle, whereas recovery was minimal in uncoated or siliconized glass tubes. A high level of radioactivity recovery was maintained for 14 days of storage of [3H]SP in triethylamine formate (TEAF) solution in BSA-precoated tubes at 4 and -20 degrees C, but decreased at 37 degrees C to less than 80% in only 3 h. Following storage in Tris-HCl (pH 7.4) buffer, a combination of HPLC and mass spectrometric analyses revealed that a significant amount of peptide bond cleavage occurred to produce the two peptides ArgProLys (RPK) and ArgProLysProGlnGln (RPKPQQ), with only a small amount of remaining intact SP. That decomposition was not observed in triethylamine formate TEAF (pH 3.14) buffer solutions.     

Haemocompatibility of endovascular coronary stents: Wiktor GX.

The stent to be examined (Wiktor-Stent, Medtronic ESTC, Kerkrade, NL) was mounted into a closed-loop tubular-system and perfused with platelet-rich plasma (PRP). As controls the tubular-system without stent (as non-thrombogenic control) and secondly the tube filled with glassbeads (as thrombogenic control) were evaluated. A decrease in the number of singularly circulating thrombocytes correlated well with an increases in circulating platelet aggregates. The increasing activation of thrombocytes was demonstrated by immunolabelling of surface structures (CD 62) which become prominent on activation of thrombocytes. The increase in case of the non-thrombogenic controls is thought to be due to the action of the roller-pump. This increase was coincident with an increase in immunologically labelled GPIIb/IIIa receptors and well correlated with an increase in platelet activation as demonstrated by the elevated CD 62 label. In spite of the use of anticoagulation principles in the perfusion model, thrombin was generated (measured by the TAT-complex) in all three cases and the completed coagulation (measured by the occurrence of fibrin D-dimers) also happened. The amount of D-dimers was small, however, in the cases of non-thrombogenic controls and of tubes equipped with stents. Only after the contact of PRP with tubes filled with glass-beads a significant increase in D-dimers followed. In conclusion the implantation of a stent led to an activation, adherence and aggregation of thrombocytes to a somewhat greater extent as in the control-system. It has, however, a much less thrombogenic surface than glass-beads.     

Genetic response ofAntirrhinum majus to acute and chronic plant irradiation

Summary Clonal lines ofAntirrhinum majus heterozygous for flower color have been used to study somatic mutations induced by irradiating plants with chronic gamma rays from cobalt-60 and acute X rays. Pink or white flecks and sectors of various sizes on the purple flowers are interpreted as resulting from mutational events at different times in flower development.Acute X-ray treatment of pre-flowering stages caused no visible change in flowers opening prior to the 15th day thereafter. On the 16th to 19th days there was dense mottling, and by the 20th or 21st day numerous eight-cell mutant spots could be counted. The number of spots induced by 1,000 r of X rays was used to obtain a preliminary estimate of an average mutation rate for the three genes studied of 6.33×10^–6 per cell per roentgen.Chronic irradiation in a gamma radiation field and greenhouse showed a linear response of mutation rate to daily dose rate, the actual rate being subject to seasonal variation. There were occasional indications of non-linearity but further tests have failed to show any significantly higher mutagenic efficiency of dose rates below 6 r/day.In plants removed from chronic gamma irradiation the frequency of mutation spots returns to the spontaneous level after about six weeks. The appearance, however, of mutant sectors on plants several months after exposure to both acute and chronic radiation indicates the induction of persistent genetic changes. Progeny tests of seeds and cuttings are planned to test the nature of these genetic events.With 6 Figures in the TextResearch carried out at the Brookhaven National Laboratory under the auspices of the U.S. Atomic Energy Commission.     

Residual radiation effect in the murine hematopoietic stem cell compartment

Stem cells surviving radiation injury may carry defects which contribute to long-term effects. The ratio of 125-iododeoxyuridine (IUdR) uptake into spleens of lethally irradiated recipient mice between day 3 and day 5 after cell transfusion revealed reduced proliferative ability (PF) of spleen seeding cells in parallel with reduced CFU-S content of donors throughout the study period of one year after 5 Gy gamma irradiation. Additional data aided in evaluating possible mechanisms of PF reduction. Within the range of the graft sizes used, PF was independent of the numbers of cells or CFU-S transfused. Radiation-induced increase in loss of label between days 3 and 5 and prolonged doubling time of proliferating cells indicated enhancement of cell maturation and increase in mitotic cycle time. Increased IUdR uptake per transfused CFU-S suggested extra divisions of transit cells due to insufficiency in the stem cell compartment. It is concluded that persisting defects in surviving stem cells interfere in a complex way with cell proliferation in the hemopoietic system.     

Effect of spleen and bone marrow regeneration on the number of hematopoietic colonies in mouse spleen]

The spleen (2/3) was removed in CBA male mice (the 1st group); in the 2nd group the bone marrow from the right posterior shin was removed. The hemopoietic splenic colonies were counted on the 8th day after the lethal irradiation and injection of 1 X 10(-6) nucleated cells of the intact spleen. A significant increase of the number of colonies in comparison with their number in control intact mice was observed. The authors suppose that this increase could also be caused by the local influence of the regenerating stroma of the spleen and by some stimulating factor discharge by the regenerating hemopoietic tissue.     

不同生长阶段遮荫对番茄光合作用、干物质分配与叶N、P、K的影响

对番茄植株做了两种不同程度的遮荫处理,观测了夏季午间遮荫对光合速率,干物质积累量及其在根,茎,叶之间的分配,和叶N,P,K的含量以及经济产量的影响,发现不同时期遮荫影响不同。（1）遮荫增加三个阶段（开花早期,盛花期和开花后期）的气孔导度和胞间CO2浓度,显著降低开花早期中午的净化合速率,但盛花期中度遮荫（40%遮荫）使净光合速率随着时间的增加逐渐上升,在开花后期表现更加明显,平均净光合速率比对照高20%以上,蒸腾速率也增加较多。（2）开花早期和盛花期重度遮荫（如本实验中的75%遮荫）显著降低根,茎的干重,而开花后期中度遮荫的根,茎干重高于对照,但遮荫对叶干重的影响不明显。（3）开花早期和盛花期遮荫不明显影响叶片中N,P,K的含量,但开花后期中度遮荫使N,P,K含量增加,（4）开花早期两种遮荫对果实产量影响较小,但盛花期重度遮荫使产量降低,全部产量中无效部分所占的比例上升,开花后期中度遮荫的总产量和有效产量增加,单果重也增加,这些结果表明,在某些时期中度遮荫可以克服夏天辐射过强,气温过高对番茄的不良影响,对番茄生长,干物质积累和提高产量等有利,在生产上有意义。     

Multiparameter analysis of CHO AL mutant populations sorted on CD59 expression after gamma irradiation

The flow cytometry mutation assay is based on detecting mutations in the CD59 gene on human chromosome 11 in CHO A(L) cells with flow cytometry, but the kinetics of mutant expression and the histogram region for mutant selection have not been studied in detail. CHO A(L) cells were analyzed by flow cytometry for CD59 expression at various times after irradiation. The mutant fraction increased to a maximum at day 6 but decreased to near background levels by day 20. Cells were sorted from six different regions on the CD59 histograms after irradiation. The growth rate was similar for cells from all regions, and the surviving fraction was 50% of that for control cells. By 14 days the CD59 expression of cells from regions 2-5 was reduced to that of region 1. Cells were also analyzed for simultaneous expression of CD59, CD44 and CD90 (all on chromosome 11) to roughly characterize the size of the mutations. Triple mutants from the sorted populations were reduced from 41% on day 6 to 8% on day 24. We conclude that the mutant region should be increased to include cells with intermediate CD59 expression; also, the loss of CD59 mutant expression over time could be explained in part by the loss of triple mutants from the population.