Expression and hemocyte-targeting of a Campoletis sonorensis polydnavirus cysteine-rich gene in Heliothis virescens larvae

The polydnavirus associated with the parasitic wasp Campoletis sonorensis is injected into the lepidopteran insect, Heliothis virescens, during parasitization, after which viral gene products suppress the cellular immune system of the hosts. Four related cysteine-rich polydnavirus gene have been identified in parasitized H. virescens larvae and grouped into a family. In this study, we investigated the expression and hemocyte targeting of the cysteine-rich VHv1.4 protein. Full-length and truncated VHv1.4 proteins were produced in a bacterial expression system, and the purified proteins were used to raise polyclonal antisera. In immunoblots the VHv1.4 protein was detected in parasitized insects as early as 6 h and throughout the entire course of parasitism. The VHv1.4 protein appeared predominantly in the plasma fraction of hemolymph from parasitized larvae, suggesting that this protein is secreted. The VHv1.4 protein expressed from a recombinant baculovirus was secreted in two lepidopteran cell lines and in larvae injected with the recombinant virus. Digestion with endoglycosidases suggests that the VHv1.4 protein is glycosylated at multiple N-glycosylation sites. Immunofluorescence assays showed that the VHv1.4 protein binds to the hemocytes, most notably the granulocytes, in H. virescens larvae. After binding, the VHv1.4 protein was internalized, probably by endocytosis. Specific binding of the VHv1.4 to granulocytes implies an important function in the suppression of host cellular encapsulation response.     

Encapsulated hemoglobin: current issues and future goals

The promise of encapsulation systems for the sequestration of hemoglobin has been the long-held belief that encapsulation more closely mimics nature's strategy for circulating hemoglobin, and could alleviate hemoglobin based toxicities and increase circulation persistence. Various polymers have been proposed to deliver hemoglobin. One approach toward the encapsulation of hemoglobin has been to employ biodegradable, biocompatible vehicles such as phospholipid vesicles, or liposomes. The majority of encapsulation work with hemoglobin over recent years has focused on liposome encapsulated hemoglobin with demonstrations of efficacy and safety in total and partial isovolemic and hypovolemic exchange models, hemodynamics, circulation persistence and organ biodistribution, processing methods, long term storage through freeze-drying, and serum changes and histopathological consequences following administration in small animals. The data collected thus far indicate that encapsulation of hemoglobin does significantly alter many of the traditionally observed effects following the administration of cell free hemoglobin solutions. Liposome encapsulated hemoglobin circulates for 20-24 hours in small animals and principally distributes to the liver and spleen. The significant accumulation of liposome encapsulated hemoglobin in these organs poses new questions for short and long-term effects on the reticuloendothelial system and macrophage function which are currently being addressed. In addition, transient hemodynamic and serum changes have been observed following the administration of liposome encapsulated hemoglobin. Many of these are similar to the effects observed following administration of liposomes without intravesicular hemoglobin and are dictated by liposome parameters such as surface charge and character, size, and lipid composition. Finally, fundamental large scale production issues such as encapsulation efficiency and particle size distribution must be optimized to facilitate the commercial development of encapsulated hemoglobin. These issues are discussed in the historical context of encapsulated hemoglobin and basic liposome research, current research status, and future challenges for the development of encapsulated hemoglobin as an artificial oxygen carrying fluid.     

Protein kinase C and casein kinase 2 phosphorylate in vitro proteins of the annexin family from eggs of loach Misgurnus fossilis

A mixture of proteins of the annexin family was obtained from the cytoplasm of mature eggs of loach Misgurnus fossilis (by reprecipitation with acid phospholipids in the presence of Ca2+). This mixture comprised five proteins with molecular weights of 58, 38, 36, 35, and 31 kD. Polyclonal rabbit antibodies against the major 31-kD protein were obtained. Western blot analysis showed that the obtained antibodies exhibit a high specificity towards the 31-kD protein from eggs and other tissues of loach and zebrafish (Brachydanio rerio). The analysis of cDNA corresponding to the 31-kD protein by screening the zebrafish cDNA library confirmed that this protein belongs to the annexin family. Phosphorylation of the obtained annexins in vitro was studied. It is shown that the 58-kD protein is phosphorylated by casein kinase 2 (CK2), whereas the 38-, 36-, 35-, and 31-kD proteins are phosphorylated by protein kinase C (PKC).     

Eggshell characteristics and penetration by Salmonella through the productive life of a broiler breeder flock

Egg weight, specific gravity, conductance, and ability of Salmonella to penetrate the shell and membranes were determined for hatching eggs from a commercial broiler breeder flock. Thirty unsanitized eggs were sampled on Weeks 29, 34, 39, 42, 48, 52, and 56 of flock age for specific gravity and conductance. An additional 10 intact eggs were inoculated with Salmonella by a temperature differential immersion method for 1 min. Eggs were then emptied of contents and filled with a selective medium that allowed visualization of Salmonella growth on the inside of the shell and membrane complex. Over the 27-wk sampling period, egg weight increased from 56 to 66 g and was positively correlated with hen age (r = 0.96, P < 0.05). However, neither specific gravity (ranging from 1.077 to 1.082) nor eggshell conductance (ranging from 14.7 to 17.9 mg weight loss/d per torr) showed any clear trend throughout the life of the flock despite the increase in egg weight. Conductance values were not correlated with specific gravity. The number of eggs positive for Salmonella penetration after 24 h incubation showed a general upward trend with flock age; however, penetration frequency and hen age were not found to be significantly correlated (P > 0.05). No relationship was found between egg specific gravity, conductance, or egg weight and the likelihood of Salmonella to penetrate the eggshell. Because shell characteristics did not change over time and the penetration patterns did vary, it is likely that factors other than specific gravity and conductance were involved in the penetration of eggshells by Salmonella.     

Xenopus laevis sperm receptor gp69/64 glycoprotein is a homolog of the mammalian sperm receptor ZP2

Little is known about sperm-binding proteins in the egg envelope of nonmammalian vertebrate species. We report here the molecular cloning and characterization of a recently identified sperm receptor (gp69/64) in the Xenopus laevis egg vitelline envelope. Our data indicate that the gp69 and gp64 glycoproteins are two glycoforms of the receptor and have the same number of N-linked oligosaccharide chains but differ in the extent of O-glycosylation. The amino acid sequence of the receptor is closely related to that of the mouse zona pellucida protein ZP2. Most of the sequence conservation, including a ZP domain, a potential furin cleavage site, and a putative transmembrane domain are located in the C-terminal half of the receptor. Proteolytic cleavage of the gp69/64 protein by a cortical granule protease during fertilization removes 27 amino acid residues from the N terminus of gp69/64 and results in loss of sperm binding to the activated eggs. Similarly, we find that treatment of eggs with type I collagenase removes 31 residues from the N terminus of gp69/64 and has the same effect on sperm binding. The isolated and purified N terminus-truncated receptor protein is inactive as an inhibitor of sperm-egg binding. Earlier studies on the effect of Pronase digestion on receptor activity suggest that this N-terminal peptide may contain an O-linked glycan that is involved in the binding process. Based on these results and the findings on the primary structure of the receptor, a pathway for the maturation and secretion of gp69/64, as well as its inactivation following fertilization, is proposed.     

Glycoform composition of serum gonadotrophins through the normal menstrual cycle and in the post-menopausal state

The heterogeneity of follicle stimulating hormone (FSH) and luteinizing hormone (LH) was investigated in five women aged 29.4 +/- 3.2 years (mean +/- SD) throughout their menstrual cycles and in five post-menopausal women aged 53.8 +/- 5.6 years. Chromatofocusing (pH range 7-4) revealed menstrual cycle stage- and postmenopausal-related differences in the serum gonadotrophin charge. There were differences in the proportion of FSH with an isoelectric point (pl) > 4.3 across phases of the menstrual cycle (P = 0.019): midcycle (MC) 50%; early to mid-follicular (EMF) 36%; late follicular (LF) 37%, luteal (L) 29% and following the menopause (PM) 17%. There was no significant difference in the proportion of LH with pl > 6.55 between midcycle (53%) and EMF, LF or L phases (36, 43 and 32% respectively); although all were greater than that found in the menopause (13%). Concanavalin A chromatography revealed less (P < 0.005) complex FSH and LH glycoforms at midcycle (63 and 13%) than in the EMF, LF and L phases (90 and 18; 90 and 20 and 93 and 24% respectively). Menopausal gonadotrophins were least complex (FSH 34%, LH 4%). There was a direct relationship between serum FSH and FSH pl/complexity, and less acidic FSH was associated with reduced FSH complexity. Increased oestradiol was associated with basic FSH isoforms during the menstrual cycle and reduced follicular phase FSH complexity. We conclude that changes in gonadotrophin glycoforms occur through the menstrual cycle which are related to changes in the prevailing steroid environment. Following the menopause oestrogenic loss resulted in acidic, relatively simple glycoforms.     

Polydnaviruses of hymenopteran endoparasitoids knock out the host insect immune response

The insect immune system reacts against invading microorganisms and parasites with the recruitment of haemocytes and with humoral response. Cellular immune reactions involve phagocytosis, nodule formation and encapsulation by different types of haemocytes whereas insect cell-free antibacterial immunity depends on the production of a number of peptides and proteins, among which lysozyme, cecropins and attacins represent the major group of immune proteins. Polydnaviruses from certain hymenopterous parasitoids interfere with both host immunity and host development. These immunosuppressive viruses exhibit an intimate genetic relationship with the parasitoid since viral sequences are integrated within the parasitoid chromosomal DNA. The viral genes expression in parasitized host induces immunosuppression and alters development of the host insect. The parasitoids developing in the host body cavity knock out the insect immune system, inducing a decline in cellular and humoral components of the immune system so that parasitoid eggs are not recognized as foreign and thereby are not encapsulated. Polydnaviruses carrying parasitoids escape the host immune response and may develop within the insect host whereas other invaders are normally destroyed by defense factors of insect haemolymph.     

Lipoproteins of the egg perivitelline fluid of Pomacea canaliculata snails (Mollusca: Gastropoda)

The lipid and protein composition of the perivitelline fluid of the eggs of Pomacea canaliculata was investigated. Two lipoproteins (PV 1 and PV 2) and one lipoprotein fraction (PV 3) were detected for the first time in gastropods. They represent 57.0, 7.5, and 35.5% of the egg total proteins, respectively. PV 1 is a glyco-carotene-protein complex with characteristics of a very high-density lipoprotein (VHDL). It has 0.33% lipids, mainly free sterols and phospholipids. The particle has a MW of 300 Kd and is composed of three subunits of 35, 32, and 28 Kd, respectively. PV 2 particle is a VHDL of 400 Kd and 3.75% lipids. The major lipid classes are free sterols and phospholipids and also have significant quantities of energy-providing triacylglycerides and free fatty acids. It is composed of two apoproteins of 67 and 31 Kd. PV 3 density corresponds to a high-density lipoprotein (HDL). It was fractionated into two subfractions "h" and "p". Fraction "h" contains 5.16% lipids, mainly free sterols, phospholipids, and free fatty acids, and two particles of 100 and 64 Kd. Dissociating electrophoresis showed two subunits of 34 and 29 Kd. Fraction "p" is composed of a single particle of 26 Kd that contains 9.5% lipids, which represents 30% of the total egg lipids. It has high levels of a carotenoid pigment. Besides it contains free fatty acids, hydrocarbons, sterified sterols, and triacylglycerides. These three fractions are probably the major supply of lipids and amino acids for the developing embryo.     

Stability in extraversion and aspects of social support at midlife

Fusion of sperm and egg plasma membranes is an early and essential event at fertilization but it is not known if it plays a part in the signal transduction mechanism that leads to the oscillations in the cytoplasmic free Ca2+ concentration ([Ca2+]i) that accompany mammalian egg activation. We have used two independent fluorescence methods and confocal microscopy to show that cytoplasmic continuity of egg and sperm precedes the onset of the first [Ca2+]i increase in mouse eggs. The Ca2+ indicator dye Ca2+-green dextran was microinjected and its transfer from egg to sperm was monitored. We found that it occurred before, and without a requirement for, any detectable [Ca2+]i increase in the egg. In separate experiments [Ca2+]i changes were recorded in populations of eggs, using fura red, and the eggs fixed at various times after some of the eggs had shown a [Ca2+]i transient. Fusion of the sperm and egg was then assessed by Hoechst dye transfer. All eggs that showed a [Ca2+]i increase had a fused sperm but more than half of the eggs contained a sperm but had not undergone a [Ca2+]i increase. These data indicate that sperm-egg fusion precedes [Ca2+]i changes and we estimate that the elapsed time between sperm-egg fusion and the onset of the [Ca2+li oscillations is 1-3 minutes. Finally, sperm-egg fusion was prevented by using low pH medium which reversibly prevented [Ca2+]i oscillations in eggs that had been inseminated. This was not due to disruption of signalling mechanisms, since [Ca2+]i changes still occurred if low pH was applied after the onset of oscillations at fertilization. [Ca2+]i changes also occurred in eggs in low pH in response to the muscarinic agonist carbachol. These data are consistent with the idea that the [Ca2+]i signals that occur in mammalian eggs at fertilization are initiated by events that are closely coupled to the fusion of the sperm and egg membranes.     

Differential regulation of P53 and Bcl-2 expression by ultraviolet A and B

The induction of apoptosis by ultraviolet (UV) radiation and other DNA damaging agents plays a critical role in monitoring the accumulation of genetic damage and the suppression of tumor development. We hypothesize that UVA and UVB induce apoptosis by modulating balances between p53 and/or bcl-2 genes. Using MCF-7 cells that express both wild-type P53 and Bcl-2 proteins, we demonstrated that UVA and UVB induced apoptosis through regulating expression of apoptosis promoting or inhibiting genes. UVA induced immediate apoptosis and downregulated bcl-2 expression. Bcl-2 expression was reduced by approximately 40% at 4 h post-150 kJ UVA irradiation per m2 with a maximum downregulation (over 70%) at 24 h. The dose-response studies revealed that significant reduction of bcl-2 expression was observed at UVA doses ranging from 50 to 200 kJ per m2; however, p53 levels were not affected by UVA. In contrast, UVB exhibited a entirely different action than UVA in that UVB substantially induced p53 expression, but had no effect on bcl-2 expression. The induction of P53 by UVB was dose and time dependent with the maximum expression at 24 h post-2 and post-4 kJ UVB irradiation per m2. Down-regulation of bcl-2 and fragmentation of DNA induced by UVA occurred earlier (approximately at 4 h) than upregulation of p53 and DNA fragmentation by UVB (12-24 h). These results suggest that UVA and UVB cause cell damage through different mechanisms and that the balances between the expression of p53 and bcl-2 may play an important role in regulating the apoptosis induced by UV irradiation.     

A simple in vitro model to study the release kinetics of liposome encapsulated material

A simple in vitro model was developed to study the release kinetics of liposome encapsulated material in the presence of biologic components. Liposomes were embedded in an agarose gel (bottom layer) formed in a glass vial and separated from the receptor compartment buffer by a second layer of agarose gel (top layer). To follow the release of liposomal contents, aqueous space markers differing in molecular weight (from 205 Dalton to 17500 Dalton) were encapsulated. The isotonic buffer in the receptor was completely changed at various time points and the amount of marker released from the agarose matrix containing the liposomes into the receptor medium determined. The release of non-encapsulated markers from the gel followed a time0.5 relationship with about 75% of a 17500 Dalton protein being released from the matrix in 48 h. In the same period, about 7% of the intact liposomes added to the agarose gel appeared in the receptor phase. The release of calcein from various liposome compositions including: (A) egg phosphatidylcholine (EPC)/egg phosphatidylglycerol (EPG) 9:1, (B) dioleoylphosphatidylethanolamine (DOPE)/cholesterylhemisuccinate (CHEMS) 2:1, and (C) dioleoylphosphatidylglycerol (DOPC)/dioleoylphosphatidylglycerol (DOPG) 2:1 was measured. Components of the biological milieu such as serum proteins and calcium influenced release of encapsulated material. This in vitro model is a convenient and reproducible system that permits the study of the release of high molecular weight molecules such as proteins from liposomal formulations in the presence of serum. It may find applications with respect to release of proteins from a variety of colloidal drug delivery systems.     

Effect of lactic acid and proteolytic enzymes on the release of organic matrix components from human root dentin

The mechanisms of organic matrix breakdown in the root caries process are not well understood. Therefore, the combined and separate effects of lactic acid and proteolytic enzymes on the degradation of human dentin collagen, glycoproteins, proteoglycans and phosphoproteins were investigated in the present study. Dentin powder was pretreated with lactic acid (pH 4.0), distilled and deionized (dd) water (pH 7.0) and EDTA/guanidine HCl (pH 7.4) for 24 h. Pellets of acid- or dd water-pretreated dentin powder were washed, dried, and then treated with trypsin, bacterial or mammalian tissue collagenase, or control buffer for 3 h. The released dentin proteins were analyzed by reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting to identify degraded type I collagen, proteoglycans, glycoproteins and phosphoproteins. All water and acid pretreatment and enzyme treatment groups demonstrated two collagen fragment bands with molecular weights at approximately 79 kD. Further studies showed that the 79 kD proteins from acid-pretreated dentin collagen were degraded by tissue collagenase, suggesting that endogenous collagenase may be involved in the degradation of root dentin collagen. Dentin proteoglycans were detectable in all the treatment groups by protein slot blotting. Relatively few distinct glycoproteins and proteoglycans, and no phosphoproteins were detected by immunoblotting. Results from this study suggest that both acids and proteolytic enzymes from either host or microbial origin are important in the degradation of human dentin matrix and the mechanisms involved in the release of various noncollagenous proteins may be different.     

Dynamic study on relationship between serum SEAIC level and hepatic pathological changes in mice infected with Schistosoma japonicum

Using purified rabbit polyclonal antibodies to SEA, avidin-biotin system and capture ELISA technique, we observed the dynamic changes in the level of the circulating soluble egg antigen-antibody complex (SEAIC) in murine sera at various weeks post infection. Simultaneously, the diameter and area of liver egg granuloma were measured by using profile analytical technique. Serum SEAIC was first detected 4 weeks post infection (p.i.), reaching peak level at 6-7th week, and then gradually dropped, and maintained at moderately high level till the end of the observation (12 weeks p.i.). Schistosome eggs appeared in liver tissue at 4 weeks p.i. No egg granuloma could be found until 6 weeks p.i. The peak of the average diameter and area of liver egg granuloma was noted at 7 weeks p.i., then dropped gradually. Its dynamic changes were consistent with that of the serum SEAIC level. It is therefore suggested that the serum SEAIC level could be a reference index reflecting the extent of the pathological changes of the liver. Moreover, SEAIC might play an important role in the pathogenesis of schistosomiasis japonica.     

Effect of dieldrin and calcium on the performance of adult Japanese quail (Coturnix coturnix japonica)

Two experiments were conducted to determine the effects of dieldrin and calcium on reproductive performance of quail. At 25% egg production the quail received diets containing 0,10 or 25 p.p.m. of dieldrin for 6, 28-day periods in experiment 1 and 0, 5, or 25 p.p.m. of dieldrin for 4, 28-day periods in experiment 2. Pesticide treatments were employed with diets containing 0.5% and 3.0% calcium. The results show that egg shell thickness, cracked eggs, egg production, feed consumption, egg weights, fertility, hatchability and body weights were not affected by dieldrin treatments. However, egg shell thickness, cracked eggs, egg production and hatchability were adversely affected by the lower calcium level. Female body weights were consistently heavier for the low calcium diet. Mortality increased in the presence of 10 and especially 25 p.p.m. of dieldrin. Livability of chicks from hens receiving rations with 10 and 25 p.p.m. of dieldrin was significantly lower than those fed no dieldrin. In summary, dieldrin was without effect on egg shell quality or other reproductive factors but did exert a detrimental effect on adult mortality and livability of progeny.     

In situ detection of AE2 anion-exchanger mRNA in the human liver

The aim of this study was to examine the effect of long-term treatment with glucocorticoids on the uterine response to oestradiol. Ovariectomized rats were treated with crystal triamcinolone acetonide (0.1 mg/100 g, i.m.) or saline (0.1 ml/100 g i.m.) for 29 days. Over this period five injections were administered, one per week. On the second day after the last triamcinolone injection, rats were treated with a single injection of oestradiol dipropionate (5 micrograms/100 g, s.c.) or vehicle (olive oil, 0.1 ml/100 g, s.c.). The effects of oestradiol in the uterus were determined by measuring mitotic index, bromodeoxyuridine (BrdU)-labelling index (BrdU was injected 2 h before the rats were killed; 2 mg/100 g, i.p.), and proliferating cell nuclear antigen (PCNA)-labelling index 24, 36 and 48 h after the injection of oestradiol or vehicle. Long-term treatment with glucocorticoids resulted in dissimilar changes in oestradiol-induced proliferation in epithelial and connective-tissue (stroma) components of the uterus. In luminal and glandular epithelia, there was an initial reduction in proliferation at 24 h, followed by an increase at 36 h and a further reduction at 48 h after the oestradiol injection. In stromal cells of the endometrium, triamcinolone treatment caused a large constant increase in oestradiol-induced proliferation throughout the experiment. The glucocorticoid treatment had no effect on the parameters without oestradiol administration.     

Worm population characteristics and pathological changes in lambs after a single or trickle infection with Teladorsagia circumcincta

The regulation of the worm population and of its pathological effects was studied after a single or trickle infection with T. circumcincta. One hundred and twenty lambs, 60 Romanov and 60 Mérinos d'Arles, 3 months old, were distributed in four balanced groups: non-infected (G0), infected with 7000 L3 per animal once and slaughtered after 4 weeks (G14) or 8 weeks (G18), and infected 8 times and slaughtered after 8 weeks (G88). Parasitological, histological, haematological parameters and weight gains were recorded on each animal. Female and artificially nursed lambs had lower worm burdens and egg counts (epg) than males and naturally suckled lambs. No difference in parasitological parameters was seen between the two breeds, but Mérinos lambs infected once, had a higher increase in pepsinogen concentrations than Romanov lambs. In the infected animals, a significant proliferation of mast and eosinophil cells was observed in the abomasum wall. Serum pepsinogen concentrations were significantly higher 3 weeks p.i. and the weight gain was depressed during the first month p.i. The worm population was more numerous and younger in group G14 compared with G18 in which 24% of the worms had been expelled during the second month p.i. The female worms in G18 were longer and had more eggs in utero and higher egg output. After the trickle infection (G88) the take was reduced, female worms were longer with more eggs in vagina (pars ejectrix) and there was a higher variability in the number of eggs (compared with G18 data). The pepsinogen rise was smaller but no specific effect was seen on histological and haematological parameters.(ABSTRACT TRUNCATED AT 250 WORDS)     

The human U4/U6 snRNP contains 60 and 90kD proteins that are structurally homologous to the yeast splicing factors Prp4p and Prp3p

Immunoaffinity-purified human 25S [U4/U6.U5] tri-snRNPs harbor a set of polypeptides, termed the tri-snRNP proteins, that are not present in Mono Q-purified 20S U5 snRNPs or 10S U4/U6 snRNPs and that are important for tri-snRNP complex formation (Behrens SE, Lührmann R, 1991, Genes & Dev 5:1439-1452). Biochemical and immunological characterization of HeLa [U4/U6.U5] tri-snRNPs led to the identification of two novel proteins with molecular weights of 61 and 63kD that are distinct from the previously described 15.5, 20, 27, 60, and 90kD tri-snRNP proteins. For the initial characterization of tri-snRNP proteins that interact directly with U4/U6 snRNPs, immunoaffinity chromatography with an antibody directed against the 60kD protein was performed. We demonstrate that the 60 and 90kD tri-snRNP proteins specifically associate with the U4/U6 snRNP at salt concentrations where the tri-snRNP complex has dissociated. The primary structures of the 60kD and 90kD proteins were determined by cloning and sequencing their respective cDNAs. The U4/U6-60kD protein possesses a C-terminal WD domain that contains seven WD repeats and thus belongs to the WD-protein family, whose best-characterized members include the Gbeta subunits of heterotrimeric G proteins. A database homology search revealed a significant degree of overall homology (57.8% similarity, 33.9% identity) between the human 60kD protein and the Saccharomyces cerevisiae U4/U6 snRNP protein Prp4p. Two additional, previously undetected WD repeats (with seven in total) were also identified in Prp4p, consistent with the possibility that 60kD/Prp4p, like beta-transducin, may adopt a propeller-like structure. The U4/U6-90kD protein was shown to exhibit significant homology, particularly in its C-terminal half, with the S. cerevisiae splicing factor Prp3p, which also associates with the yeast U4/U6 snRNP. Interestingly, U4/U6-90kD shares short regions of homology with E. coli RNase III, including a region encompassing its double-stranded RNA binding domain. Based on their structural similarity with essential splicing factors in yeast, the human U4/U6-60kD and 90kD proteins are likely also to play important roles in the mammalian splicing process.     

Effect of hen production age on egg composition and embryo development in commercial Pekin ducks

At 26, 31, 36, and 42 wk of age, eggs were collected from the same duck breeder flock to study the effects of hen production age on egg composition and embryo development. At each hen age, yolk and albumen measurements were made on a random subsample of unincubated eggs. Embryo and yolk sac measurements were made at 20, 22, 24, 26, 27, and 28 d of incubation. Egg weight was significantly affected by hen production age, but most of the observed age effects occurred between 26 and 31 wk with minimal age differences thereafter due to a quantitative feed restriction for egg weight. Yolk weight increased significantly with hen production age, with the largest increase, 6.6 g, also occurring between 26 and 31 wk of age. Yolk-free duckling weight increased with hen production age at 27 d of incubation. The yolk sacs of embryos from the 31-, 36-, and 42-wk-old hens were heavier at 20 and 22 d, before the differentiation in embryo weight. Differences in yolk sac weight were not consistently affected by hen production age between 26 and 28 d of incubation. The eggs from the 42-wk-old hens weighed 3.3 g less than those from the 31-wk-old hens, but yolk-free duckling weight at 27 d was 2.9 g heavier from the 42-wk-old hens.     

Tn4351-generated non-haemolytic and/or non-pigmented mutants of Porphyromonas gingivalis

Escherichia coli-Porphyromonas gingivalis plasmid-shuttle vectors R751::* omega 4, pVOH1, and pVAL-1 were used for isolation of non-pigmented, defective protease, or non-haemolytic activity phenotypes in P. gingivalis. Transfer frequencies for R751::* omega 4, pVOH1, and pVAL-1 varied from 1.7 x 10(-9) to 5.3 x 10(-11) depending on the P. gingivalis 381 and ATCC 33277 strains. Two erythromycin-resistant transconjugants were screened from P. gingivalis 381 and eighteen were screened from ATCC 33277. Among isolated transconjugants, two from ATCC 33277 contained only one transposon insertion, while others included both Tn4351 and R751 sequences. The one transconjugant which contained a single insertion of Tn4351, designated NUM-T14, demonstrated defective pigmentation, and defective protease and haemolytic activities. The other transconjugant, designated NUM-T29, demonstrated defective haemolytic activity, but had black pigmentation and protease activity. Cell surface protein analysis by SDS-PAGE indicated that 40 and 18 kD proteins were lost or reduced and that 45 and 36 kD proteins were made to appear in both NUM-T14 and NUM-T29 transconjugants.