Immunolatex visualisation of cell surface Forssman and polyamine antigens

Specific antibodies were chemically linked to latex spheres. These immunolatex spheres, which are stable on storage for up to 4 weeks at 4°C have been used to visualise the corresponding antigens on chick embryo cells under the scanning electron microscope. The Forssman antigen has been successfully visualised using the indirect method and the polyamine antigen by the direct method. Using the immunolatex technique, visualisation of Forssman antigenic sites was achieved with a dilution of antiserum 80-fold greater than that necessary for a positive reaction using the immunofluorescence technique.     

New immunolatex spheres: visual markers of antigens on lymphocytes for scanning electron microscopy

New immunochemical reagents consisting of antibodies bound to small latex spheres were used as visual markers for the detection and localization of cell surface antigens by scanning electron microscopy. Cross-linked latex spheres of various sizes from 300 to 3,4000 A in diameter were synthesized by aqueous emulsion copolymerization of methacrylate derivatives containing hydroxyl and carboxyl functional groups. Proteins and other molecules containing primary amino groups were covalently bonded to the acrylic spheres under a variety of mild conditions by the aqueous carbodiimide, cyanogen bromide, and glutaraldehyde methods. For use in the indirect immunochemical-labeling technique, goat antibodies directed against rabbit immunoglobulins were bonded to the spheres. These immunolatex reagents were shown to bind only to cells (red blood and lymphocytes) which had previously been sensitized with rabbit antibodies against cell surface antigens. Mouse spleen lymphocytes with exposed immunoglobulins on their surface (B cells) were labeled with these spheres and distinguished from unlabeled or T lymphocytes by scanning electron microscopy. The distribution of Ig receptors on lymphocytes was also studied using the spheres as visual markers. When lymphocytes were fixed with glutaraldehyde and subsequently labeled with the immunolatex reagents, a random distribution was observed by scanning electron microscopy; a patchy distribution was observed when unfixed lymphocytes were used. These results are consistent with studies using ferritin-labeled antibodies (S. De Petris and M. Raff. 1973. Nature [Lond.]. 241:257.) and support the view that Ig receptors on lymphocytes undergo translational diffusion. In addition to serving as visual markers for scanning electron microscopy, these latex spheres tagged with fluorescent or radioactive molecules have applications as highly sensitive markers for fluorescent microscopy and as reagents for quantitative studies of cell surface antigens and other receptors.     

Host recognition in the Rhizobium-soybean symbiosis

Polar binding of Rhizobium japonicum to roots and root hairs of Glycine soja (L.) Sieb. and Zucc. is specifically inhibited by d-galactose and N-acetyl-d-galactosamine, haptens of Glycine max seed lectin. A protein, immunologically cross-reactive with the G. max seed lectin, is present in G. soja seed extracts. Peptide mapping of the purified G. max and G. soja lectins indicates that the two are similar in structure. Soybean lectin can be localized on the surface of both G. max and G. soja roots by indirect immunolatex techniques. These observations indicate that the Rhizobium-binding lectin, previously isolated from seeds, also is present on the root surface-the site of the initial steps in the infection. This lectin is capable of binding Rhizobium japonicum to the root.     

Protein-bound polyamines in the plasma of mice grafted with the Lewis lung carcinoma

Protein-bound polyamines were isolated from the plasma of mice using antipolyamine antibodies covalently linked to magnetic latex spheres. Their subsequent separation by polyacrylamide gel electrophoresis (PAGE) showed that in plasma from normal mice, 3 proteins (27, 55 and 82 kDa) carrying polyamines could be visualized, whereas in mice bearing the Lewis lung carcinoma at least 8 other proteins of higher molecular mass (5 of 94, 110, 130, 145 and 160 kDa, and 3 of greater than 170 kDa) had bound polyamines. These protein-bound polyamines could be detected from the first week after tumour graft; they increased during the second and third week but decreased thereafter. These proteins were not bound by immunolatex spheres preincubated with spermine bound to a protein-carrier insulin. Moreover, the appearance of these protein-bound polyamines was not a consequence of the inflammatory process since in mice infected with heat-inactivated Brucella abortus, with the exception of a 65 kDa protein, polyamines were bound to the same proteins found in normal mice. In mice grafted with the Lewis lung carcinoma the concomitant decrease in transglutaminase-mediated polyamine (e.g. putrescine) binding capacity of plasma proteins provides additional evidence for the presence in vivo of polyamines already bound to plasma proteins.     

Absorption and metabolism of purines by the lower intestine of the chicken.

1. Absorption of purines and their metabolism by the lower intestine were estimated by using the everted gut sacs from the colo-rectum and caecum of the chicken. 2. Adenine, hypoxanthine and uric acid were appreciably absorbed from the colo-rectum and caecum, and an especially high rate was observed in the absorption of uric acid from the colo-rectum. 3. Guanine was not absorbed unchanged from either the colo-rectum or the caecum and a small amount of xanthine was absorbed only from the caecum. 4. Hypoxanthine was also absorbed in uric acid form, to a much lesser extent, in xanthine form from the colo-rectum and caecum, adenine and xanthine in uric acid form from the colo-rectum and adenine in hypoxanthine form from the colo-rectum and caecum. 5. Adenine was metabolized to hypoxanthine and xanthine, guanine and hypoxanthine to uric acid and xanthine, and xanthine to adenine, in both mucosal fluids of the colo-rectum and caecum. The conversion of guanine to uric acid in the caecum was most active, being almost twice as much as that in the colo-rectum.     

An insect lipid transfer particle promotes lipid loading from fat body to lipoprotein

The role of Manduca sexta lipid transfer particle (LTP) in the transport of lipid from fat body to lipophorin was investigated in vitro. Fat body that contained radiolabeled lipid was incubated with either high density lipophorin or low density lipophorin, and it was shown that lipid was transferred from fat body to lipophorins. The transfer of diacylglycerol was blocked by preincubating fat body with LTP antibody. Furthermore, transfer was restored by the addition of LTP, indicating that LTP promotes the transfer of lipid from fat body to lipophorins. Using lipophorins radio-labeled in their lipid moiety, transfer of lipid from lipophorin to fat body was demonstrated. This transfer was not mediated by LTP. The adipokinetic hormone induced diacylglycerol mobilization from the fat body and the concomitant interconversion of high density lipophorin to low density lipophorin were performed in vitro and were shown to require the presence of LTP.     

Body height in relation to rural-urban migration in Poland

A survey was conducted among 2800 students studying in Wroc?aw, Poland. The questionnaire included questions on the body height of the students and their parents, and place of residence and migration patterns of the students themselves, their parents and their grandparents. Body height in both students and their parents was positively correlated with the size of their place of residence. This was particularly true for male students and their fathers. Body height in students and parents from mobile families was not significantly different from that of their peers from non-mobile families. Body height in mobile individuals was generally between that of non-mobile individuals from rural areas and that of non-mobile individuals from large urban centres. Students from families that had migrated from smaller urban centres to larger ones were taller than students from families that had migrated from rural areas to urban centres. Body height in students was also correlated with the kind of migration that took place. In the students' mothers, body height was higher if the maternal grandparents moved from smaller urban centres to larger urban centres than if the maternal grandparents moved from rural areas to urban centres. In female students, body height depended on whether their mothers had migrated from smaller places of residence to larger places of residence, but was not affected by the degree of migration. Intra-generational migration during the generation of the students' grandparents was associated with increased body height in the students' mothers. On the other hand, intergenerational migration during the generations of the students' grandparents and parents was associated with increased body height in the students' fathers and in female students. Body height was not a reliable indicator of whether an individual migrated from rural areas to Wroc?aw. Far more reliable indicators were the size of the place the student lived their whole life and whether the family had lived in an urban environment for at least two generations.     

Quantitation of lectin binding sites in human colon mucins by use of peanut and wheat germ agglutinins

We have developed a novel method for quantitation of lectin binding sites in mucins derived from colon tissues. Binding of peanut agglutinin and wheat germ agglutinin was measured in extracts from normal and malignant human colon epithelium. Binding of wheat germ agglutinin was used as an estimate of the total mucin present in the tissue extract. Peanut agglutinin was found to bind to mucin from normal colon, but at levels that may be difficult to appreciate by fluorescence microscopy. The yield of mucin extracted from colon cancer was more variable than that from normal colon, and the binding ratio of peanut agglutinin to wheat germ agglutinin was greater in extracts from tumors than in normal tissues. Our findings confirm the histological observation that peanut agglutinin binds more avidly to mucins from colon cancer than to those from normal colon. The finding of peanut agglutinin binding sites in mucins front normal colon was not expected. The quantitative technique may have detected small numbers of binding sites not readily appreciable by fluorescence microscopy. Alternatively, the chromatographic method for measuring lectin binding may be sufficiently sensitive to detect nonspecific binding of the lectin to terminal galactose residues other than the Thomsen-Friedenreich antigen.     

Cyclic adenosine monophosphate (cAMP) production in avian theca cells during follicular maturation

LH was used to stimulate cAMP production in theca cells from the 5 largest preovulatory follicles of hens and this was related to LH-stimulated androstenedione production in the same cells. cAMP production was stimulated by LH to the same extent in theca cells from each follicle. However, LH was not effective in stimulating androstenedione production in theca cells from the largest follicle (T1), although androstenedione production was greatly increased by LH in the smaller follicles (T2-T5). Effects similar to those of LH on cAMP production were observed in response to forskolin, indicating that the intrinsic adenylate cyclase activity was similar in theca cells from each follicle. In addition, forskolin was unable to stimulate androstenedione production by T1 cells. Our results provide evidence that the levels of receptor-mediated and non-receptor-mediated cAMP production are similar in theca cells from the 5 largest follicles. We conclude that the step that restricts the ability of T1 cells to produce androgen is distal to cAMP generation.     

Mineralization of copper, manganese and zinc from rock mineral flour and city waste compost for efficient use in organic farming

Restricted supply of micronutrients is a common constraint for plant growth worldwide, especially in organic farming systems where nutrients supply to crops mostly depends on the mineralization of native soil organic matter, decomposition of applied manures and crop residues. A laboratory incubation study was therefore conducted to investigate the potential release of copper (Cu), manganese (Mn) and zinc (Zn) from the rock mineral flour (RMF) and city waste compost (CWC) as compared to inorganic micronutrient fertilizers for 140 days. Release of the micronutrients from RMF and CWC showed different trends. The results showed that about 4.6% of Cu added as RMF was released irrespective of the quantity of the RMF applied. However, Cu release from CWC increased from 0.7 to 3.5% as the amount of compost added was increased. Copper recovery from copper sulphate was 98%. Manganese release from RMF decreased from 114 to 103% as the RMF level was increased, while the corresponding decrease in Mn release from CWC was from 14 to ?3%. Manganese recovery from manganese sulphate was 100%. Zinc release from RMF increased from 5.8?15.5%, with an increase in the amount of RMF applied, while no Zn was released from CWC. Recovery of Zn from zinc sulphate was 98%. These results show that RMF and CWC could be used to meet Cu, Mn and Zn requirements of organically grown cereals. The results of the investigation have general applicability in organic farming.     

Swine vesicular disease viruses isolated from healthy pigs in non-epizootic period. I. Isolation and identification

A total of 524 fecal samples were collected from healthy swine of 36 hog farms scattered all over Japan from which had been detected neutralizing antibody against swine vesicular disease (SVD) virus. A virus was isolated from 21 of them. It was neutralized by antiserum against SVD virus. Of the 21 samples, 19 were derived from six farms, in areas which remained to be free from SVD in the past years and two from a farm where SVD broke out 18 months before. The cross-neutralization test was carried out with seven strains of SVD virus isolated from healthy pigs (SVDV-H) and five strains of SVD virus isolated from diseased pigs (SVDV-D). There was, however, no significant difference in antigenicity between the two groups. Some strains of SVDV-H were antigenically close to the Faulkner strain of Coxsackie B5 virus, and others to the freshly isolated strain of this virus. Neutralizing antibody of low titer against SVD virus was detected from pigs kept in areas free from SVD. It was presumed to have been produced in these pigs involved in silent infection with this virus.     

Sex steroid content and metabolism in Trichostrongylus colubriformis (Nematoda)

Testosterone, progesterone and cholesterol were found in mixed sexes of the nematode Trichostrongylus colubriformis from goats, according to thin-layer, gas-liquid and high-performance liquid chromatography. The structure of these steroids was confirmed by proton nuclear magnetic resonance and mass spectroscopy. Melting points of the worms' steroids were similar to authentic standards of the steroids. Estradiol was not detected in worms from either goat sex. Cholesterol was about 0.08% of the worms' dry weight in helminths from either sex of host. Testosterone was 0.02% of the dry weight when worms were taken from male goats, but only 0.005% from female goats. Progesterone was not detected in worms from male goats, but was 0.005% of the dry weight of helminths from female hosts. Incubation of a worm preparation with tritiated steroids showed that progesterone was converted to 17-alpha-hydroxyprogesterone, based on retention during radioactive thin-layer and gas-liquid chromatography, and co-crystallization. Testosterone, cholesterol and 17-beta-estradiol were not metabolized.     

Survival and feeding responses of Anacanthotermes ochraceus (Hodotermitidae: Isoptera) to local and imported wood

Forty-six local and imported wood were tested for resistance to feeding damage by the termite Anacanthotermes ochraceus (Burmeister), the most dominant species in the United Arab Emirates and the Arab Gulf region. Wood was used for construction, wall paneling, and furniture. Wood was evaluated in a 4-wk forced feeding bioassay. Each wood block was graded by the amount of termite damage by using a damage rating index (DRI) of 0 to 5 and wood rating index from very resistant to very susceptible wood. Local wood was mostly susceptible to feeding of termites; imported wood varied in resistance to feeding damage. Wood was placed in groups according to the percentages of weight loss (WL), termite survival (TS), and DRI. Wood was classified as very resistant (%WL from 0.0 to 0.3, %TS from 0.01 to 0.5, and DRI of 0.01), resistant (%WL from 1.1 to 4.9, %TS from 0.8 to 4.8, and DRI of 1.0), moderately resistant (%WL from 6.6 to 9.3, %TS from 6.3 to 8.3, and DRI of 2.0-2.3), slightly resistant (%WL from 10.1 to 19.9, %TS from 9.5 to 28.0, and DRI of 2.5-3.5), susceptible (%WL from 21.5 to 48.6, %TS from 37.3 to 64.8, and DRI of 4.0-4.3) and very susceptible (%WL from 50.0 to 59.8, %TS from 72.8 to 79.0, and DRI of 4.5-5.0). The characterization of the extracts of resistant wood may prove of economic value and lead to the development of new chemicals (repellents or antifeedants) for termite control.     

Influence of ethanol on the liver metabolism of fed and starved rats

1. The influence of ethanol on the metabolism of livers from fed and starved rats has been studied in liver-perfusion experiments. Results have been obtained on oxygen consumption and carbon dioxide production, on glucose release and uptake by the liver and on changes in the concentrations of lactate and pyruvate and of β-hydroxybutyrate and acetoacetate in the perfusion medium. 2. Oxygen consumption and carbon dioxide production were lower in livers from starved rats than in livers from fed rats. Ethanol had no effect on the oxygen consumption of either type of liver. After the addition of ethanol to the perfusion medium carbon dioxide production ceased almost completely, the change being faster in livers from starved rats. 3. With livers from fed rats glucose was released from the liver into the perfusion medium. This release was slightly greater when ethanol was present. With livers from starved rats no release of glucose was observed, and when ethanol was added a marked uptake of glucose from the medium was found. A simultaneous release of glycolytic end products, lactate and pyruvate, into the medium occurred. 4. Acetate was the main metabolite accumulating in the perfusion medium when ethanol was oxidized. With livers from starved rats a slightly increased formation of ketone bodies was found when ethanol was present. 5. The lactate/pyruvate concentration ratio in the perfusion medium increased from 10 to 87 with livers from fed rats and from 20 to 171 with livers from starved rats when the livers were perfused with ethanol in the medium. The β-hydroxybutyrate/acetoacetate concentration ratio increased from 0·8 to 7·6 with livers from fed rats and from 1·0 to 9·5 with livers from starved rats when ethanol was added to the medium. 6. The effects of ethanol are discussed and related to changes in the redox state of the liver that produce new conditions for some metabolic pathways.     

Products of phospholipid metabolism in Bacillus stearothermophilus.

The products of phospholipid turnover in Bacillus stearothermophilus were determined in cultures labeled to equilibrium and with short pulses of [32P]phosphate and [2-3H]glycerol. Label lost from the cellular lipid pool was recovered in three fractions: low-molecular-weight extracellular products, extracellular lipid, and lipoteichoic acid (LTA). The low-molecular-weight turnover products were released from the cells during the first 10 to 20 min of a 60-min chase period and appeared to be derived primarily from phosphatidylglycerol turnover. Phosphatidylethanolamine, which appeared to be synthesized in part from the phosphatidyl group of phosphatidylglycerol, was released from the cell but was not degraded. The major product of phospholipid turnover was LTA. Essentially all of the label lost from the lipid pool during the final 40 min of the chase period was recovered as extracellular LTA. The LTA appeared to be derived primarily from the turnover of cardiolipin and the phosphatidyl group of phosphatidylglycerol. Three types of LTA were isolated; an extracellular LTA was recovered from the culture medium, and two types of LTA were extracted from membrane preparations or whole-cell lysates by the hot phenol-water procedure. Cells contained 1.5 to 2.5 mg of cellular LTA per g of cells (dry weight), over 50% of which remained associated with the membrane when cells were fractionated. Over 75% of the 3H label incorporated into the cellular LTA pool during a 90-min labeling period was released from the cells during the first cell doubling after the chase. Label lost from the lipid pool was incorporated into cellular LTA which was then modified and released into the culture medium.     

Study of fertilising capacity of spermatozoa after heterospermic insemination in rabbit using DNA markers

The aim of this study was to evaluate the fertilising capacity of males belonging to a rabbit line selected for growth rate using heterospermic insemination and genetic markers. Semen from five males was used to make pools of three of them, and to perform homospermic insemination. Insemination was carried out in receptive multiparous lactating does with 6 million spermatozoa per insemination dose. DNA from 360 young rabbits born from heterospermic insemination, 5 sires and 42 does were amplified to nine microsatellite loci for determination of the offspring rate per male. Although each female was inseminated with the same number of spermatozoa from each male (2 million from a total dose of 6 million), sperm from one male was always dominant, notable differences being observed in the offspring among the males with similar semen quality (83-68% from dominant male versus 31-0% from non-dominant, P<0.05 ).     

Crystallographic analysis of the thermal motion of the inclusion complex of cyclomaltoheptaose (beta-cyclodextrin) with hexamethylenetetramine

The crystal structure of the inclusion complex of cyclomaltoheptaose (beta-cyclodextrin) with hexamethylenetetramine was determined at temperatures of 123, 173, 223, and 293 K. The rigid-body motion of the host and guest molecules was evaluated by means of the TLS method that represents the molecular motion in terms of translation, libration, and screw motion. In increasing the temperature from 123 to 293 K, the amplitude of the rigid body vibration of the host molecule was increased from 1.0 to 1.3 degrees in the rotational motion and from 0.16 to 0.17 A in the translational motion. The cyclomaltoheptaose molecule has the flexibility in seven alpha-(1-->4)-linkages, and each glucose unit was in the rotational vibration around an axis through two glycosidic oxygen atoms. As a result, the rigid-body parameters of cyclomaltoheptaose were considered to be overestimated because of including the contribution from the local motion of glucose units. In contrast, for the guest molecule having no structural flexibility, the TLS analysis demonstrated that the atomic thermal vibration was mostly derived from the rigid body motion. The rotational amplitude of hexamethylenetetramine was changed from 5.2 to 6.6 degrees in increasing the temperature from 123 to 293 K, while the change of the translational amplitude was from 0.20 to 0.23 A. The translational motion of the guest molecule was hindered by the inside wall of the host cavity. The molecular motion was characterized by the rotational vibration around the axis through two nitrogen atoms that were involved in the hydrogen-bond formation.     

Antimicrobial susceptibilities of enterococci isolated from faeces of broiler and layer chickens

Enterococci were isolated from faecal droppings of chickens in broiler and layer farms and the susceptibilities to nine therapeutic antimicrobial agents and six growth-promoting antibiotics were determined by the agar dilution method. Resistance to therapeutic antimicrobial agents such as ampicillin, clindamycin, erythromycin, streptomycin, tetracycline or tylosin was more frequent in enterococcal isolates from broiler farms than in those from layer farms. Resistance to ofloxacin was rare, occurring in only one (0.7%) of the Enterococcus faecium isolates from broiler farms. Resistance to growth-promoting antibiotics such as avilamycin, salinomycin and virginiamycin was common among isolates from broiler farms. Of the E. faecium isolates from broiler farms, 12.4% were resistant to avilamycin and 27.4% were resistant to virginiamycin. Resistance to salinomycin was detected in all enterococcal species, ranging from 12.4% of E. faecium isolates to 50% of E. hirae isolates.     

蠊卵荚内寄生啮小蜂的生物学

本文报道了寄生于烟色大蠊Periplaneta fuliginosa(Serville)卵荚内的两种啮小蜂:Tetrastichus hagenowii和Tetrastichodes sp.,并讨论了Tetrastichus hagenowii的生物学特性。所收集的(虫非)蠊卵荚中,该蜂的自然寄生率为50.38%。观察14枚卵荚的出蜂数,最多为202只/枚,最少为10只/枚,平均为74.92只/枚。雌雄性比为1:0.42。该蜂4月上旬开始出现,5—8月是繁殖高蜂期,11月上旬则少见。接蜂的8枚卵荚孵出的成蜂平均存活12.8天(14.4天.10.2天)。蜂在卵荚内的发育时间与温度有关,在夏季比在秋冬季显著为短。成蜂的产卵期较长:而且除产卵于新鲜(虫非)卵荚内,也产卵于已产出7天以上的卵荚内。本文的观察结果为人工繁殖该啮小蜂用以防治室内(虫非)蠊提供了科学依据。