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Comparative study of three different membranes for guided bone regeneration of rat cranial defects.

The purpose of the study was to compare the efficacy of a non-resorbable polytetrafluoroethylene (e-PTFE) membrane (Gore-Tex) versus a resorbable polyglactin membrane (Vicryl) and a newly designed collagenic membrane for enhancing bone regeneration on rat skull defects. The study was conducted on 30 adult Wistar rats. On each animal, two symmetrical, 6 mm wide, full-thickness, skull defects were created in the parietal regions. The right defect was chosen as the experimental site and the left one was left empty as a control. Each experimental site was covered by an inner and outer membrane. The 30 rats were divided into three groups: In group 1 (n=10), a non-resorbable polytetrafluoroethylene (e-PTFE) membrane was used. In group 2 (n= 10), a resorbable polyglactin 910 membrane was used. In group 3 (n= 10), a collagen membrane processed from avian eggshell was used. In each group, the animals were euthanized at 60 days. The harvested specimens were processed for contact radiography and standard histological examination. The results were assessed by a Fisher's exact test. In group 1, partial bone healing was observed in seven out of 10 animals and complete in three out of 10 animals (P<0.001). In group 2, no or minimal bone healing was observed in seven out of 10 animals and partial bone healing was observed in three out of 10 animals. In group 3, no or minimal bone healing was observed in nine out of 10 cases and partial bone healing in only one animal. In conclusion, only the non-resorbable e-PTFE membrane group exhibited a favourable result in this study. This study suggests that the structure of the membrane is at least as important as its composition.

Absorbable Implants↗

Spoon retractors for soft mass.

We have designed spoon retractors for removing a soft mass. The retractor is composed of a main part which is eggshell like and concave in shape, with sizes varying from 5 mm to 4 cm in width, and a flexible shaft. The retractor enables retraction to hold a soft mass upward creating an ample space for dissecting it from surrounding structures in the depth. The spoon retractor has proven to be useful for gliomas and metastatic tumours, particularly for haemorrhagic lesions such as haemangioblastomas and arteriovenous malformations, because the retractor facilitates an en bloc removal of the mass by retracting it without causing laceration or rupture, and with feeding or draining vessels easily visualised.

Brain Neoplasms↗

A study of the hepatic lipids during chick embryo development. Effect of triiodothyronine.

The content of hepatic phospholipids, cholesterol and triglycerides was studied in 14, 17 and 20 day old chick embryos and newborn chicks (21 incubation days). An increase in the total lipid content and in the other lipidic fractions throughout embryo development is observed, mainly between 17 and 20 days of incubation, remaining practically constant until the hatchability day. Nevertheless a sharp depletion of triglycerides in broken eggshell day is revealed. Effect of a single dose of 3-3'-5-Triiodothyronine (T3) on hepatic lipid levels was determined. Throughout embryonary period, the hormone only affects triglyceride levels enhancing its biosynthesis in the 20th incubation day. All the other variations are described in newborn animals.

Animals↗

The SQV-1 UDP-glucuronic acid decarboxylase and the SQV-7 nucleotide-sugar transporter may act in the Golgi apparatus to affect Caenorhabditis elegans vulval morphogenesis and embryonic development.

Recent findings indicate that glycosaminoglycans can play important roles in animal development. The genes sqv-3, -7, and -8, which are necessary for vulval morphogenesis in Caenorhabditis elegans, affect the biosynthesis of chondroitin and heparan sulfate glycosaminoglycans. We cloned sqv-1 and showed that the SQV-1 protein is a type II transmembrane protein that functions as a UDP-glucuronic acid decarboxylase. SQV-1 localizes to punctate cytoplasmic compartments and colocalizes with the SQV-7 nucleotide-sugar transporter, which probably acts in the Golgi apparatus. SQV-1 and SQV-7 are both expressed in the vulva and in oocytes, where they likely act in vulval morphogenesis and embryonic development, respectively. Progeny of sqv-7 and sqv-1 null mutants fail to initiate cytokinesis, possibly because they are unable to separate the plasma membrane from the eggshell, a defect analogous to that of incomplete vulval invagination.

Amino Acid Sequence↗

Probable extirpation of a breeding colony of Short-tailed Albatross (Phoebastria albatrus) on Bermuda by Pleistocene sea-level rise.

Albatrosses (Diomedeidae) do not occur in the North Atlantic Ocean today except as vagrants, although five species were present in the early Pliocene. No fossil breeding sites of albatrosses were known previously. The timing of extinction of albatrosses in the North Atlantic was likewise unknown. Deposits that formed near present-day sea level along the southeastern shore of Bermuda contain remains of a former breeding colony and include intact eggshells and bones of embryos, juveniles, and adults of Short-tailed Albatross (Phoebastria albatrus), a critically endangered species now confined to a few islets in the northwestern Pacific Ocean. These deposits are correlated with the middle Pleistocene Lower Town Hill Formation, which at other sites have a radiometric age of 405,000 years ago. This equates with the marine isotope stage 11 interglacial, which culminated in a rise in sea-level to >+20 m. Bones of a juvenile Short-tailed Albatross were also found in beach deposits at +21.3 m from this same interglacial. We interpret the extirpation of albatrosses on Bermuda as probably resulting from lack of nesting sites protected from storm surges over the little emergent land that remained at the height of the marine isotope stage 11 sea level rise.

Altitude↗

Purification of a family of specific messenger ribonucleic acids from moth follicular cells.

When moth follicular cells synthesize the characteristic, low-molecular-weight chorion (eggshell) proteins, they contain at least two groups of 8-9S RNAs which appear to be chorion mRNA. This class of RNA yields a characteristic radiolabeled profile on polyacrylamide slab gels. The same profile is obtained irrespective of whether lebeling is performed in vivo or in organ culture, and whether the RNAs are purified from whole cells or specifically released from polysomes. Polysomes contain putative chorion mRNA only when the cells actually synthesize chorion protein. The RNA size is correlated with the size of the chorion proteins: two month species differing in the size distribution of their chorion proteins show a corresponding difference in the size distribution of the RNAs.

Animals↗

Silkmoth chorion proteins: sequence analysis of the products of a multigene family.

Five polypeptide components have been isolated from the eggshell (chorions) of a silkmoth. Two are homogeneous on sodium dodecyl sulfate and isoelectric focusing gels, and three contain predominantly two proteins each. Amino acid analyses show that all five components are similar to each other. These proteins have been sequenced from the amino terminus. Homogeneous components yielded single sequences; heterogeneous components yielded two residues at some positions, consistent with their containing two major electrophoretic components. Striking similarities are apparent among all these sequences. These similarities can be increased dramatically by separating each of the three protein mixtures into two sequences and introducing a small number of gaps or insertions. This is due in part to bringing into register a portion that contains short repeating subunits found in all sequences. All proteins are also characterized by a region of high cysteine content near the amino terminus followed by a longer low-cysteine region. The data suggest that these proteins share a common evolutionary origin and are encoded by a multigene family.

Amino Acid Sequence↗

A short 5'-flanking DNA region is sufficient for developmentally correct expression of moth chorion genes in Drosophila.

Fusions with the bacterial gene for chloramphenicol acetyltransferase followed by P-element-mediated germ-line transformation in Drosophila have permitted localization of the DNA sequence that confers a high degree of developmental specificity on a pair of silkmoth eggshell (chorion) genes. The short, 272-base-pair, 5'-flanking region shared by the divergently transcribed genes is sufficient for developmentally appropriate expression when placed upstream of the chloramphenicol acetyltransferase gene, in either orientation. A highly conserved motif within that region, TCACGT, is essential for chorion-specific expression.

Animals↗

The gastrulation defective gene of Drosophila melanogaster is a member of the serine protease superfamily.

The establishment of dorsal-ventral polarity in the oocyte involves two sets of genes. One set belongs to the gurken-torpedo signaling pathway and affects the development of the egg chorion as well as the polarity of the embryo. The second set of genes affects only the dorsal-ventral polarity of the embryo but not the eggshell. gastrulation defective is one of the earliest acting of this second set of maternally required genes. We have cloned and characterized the gastrulation defective gene and determined that it encodes a protein structurally related to the serine protease superfamily, which also includes the Snake, Easter, and Nudel proteins. These data provide additional support for the involvement of a protease cascade in generating an asymmetric signal (i.e., asymmetric Spätzle activity) during establishment of dorsal-ventral polarity in the Drosophila embryo.

Amino Acid Sequence↗

Molecular cloning and characterization of lustrin A, a matrix protein from shell and pearl nacre of Haliotis rufescens.

A specialized extracellular matrix of proteins and polysaccharides controls the morphology and packing of calcium carbonate crystals and becomes occluded within the mineralized composite during formation of the molluscan shell and pearl. We have cloned and characterized the cDNA coding for Lustrin A, a newly described matrix protein from the nacreous layer of the shell and pearl produced by the abalone, Haliotis rufescens, a marine gastropod mollusc. The full-length cDNA is 4,439 base pairs (bp) long and contains an open reading frame coding for 1,428 amino acids. The deduced amino acid sequence reveals a highly modular structure with a high proportion of Ser (16%), Pro (14%), Gly (13%), and Cys (9%). The protein contains ten highly conserved cysteine-rich domains interspersed by eight proline-rich domains; a glycine- and serine-rich domain lies between the two cysteine-rich domains nearest the C terminus, and these are followed by a basic domain and a C-terminal domain that is highly similar to known protease inhibitors. The glycine- and serine-rich domain and at least one of the proline-rich domains show sequence similarity to proteins of two extracellular matrix superfamilies (one of which also is involved in the mineralized matrixes of bone, dentin, and avian eggshell). The arrangement of alternating cysteine-rich domains and proline-rich domains is strikingly similar to that found in frustulins, the proteins that are integral to the silicified cell wall of diatoms. Its modular structure suggests that Lustrin A is a multifunctional protein, whereas the occurrence of related sequences suggest it is a member of a multiprotein family.

Amino Acid Sequence↗

Identification of a cDNA encoding a retinoid X receptor homologue from Schistosoma mansoni. Evidence for a role in female-specific gene expression.

Schistosoma mansoni, a multicelluar eukaryotic blood fluke, is a major cause of morbidity worldwide in humans. The study of female parasite growth, development, and gene regulation is important because the eggs produced are responsible for the pathogenesis observed in schistosomiasis. p14, an eggshell precursor gene expressed only in sexually mature females in response to a male stimulus, is a model for female-specific gene regulation. The upstream region of the p14 gene shares sequences present in insect genes known to be regulated in a sex-, temporal-, and tissue-specific manner by members of the steroid receptor superfamily. Herein, we report the identification and characterization of a cDNA that encodes the S. mansoni (Sm) RXR homologue. Sequence analysis predicts and Western blot analysis confirms the synthesis of a 74-kDa protein, the largest member of the RXR family reported to date. We show by electrophoretic mobility shift assay analysis that SmRXR binds to cis-elements of the p14 gene including a direct repeat that follows the "3-4-5" rule of binding elements recognized by members of the steroid receptor superfamily. Furthermore, we demonstrate that SmRXR can act as a transcription activator in the yeast one-hybrid system. Through quantitative reverse transcriptase-polymerase chain reaction, we show that the SmRXR gene is constitutively expressed and thus must play multiple roles throughout the schistosome life cycle.

Amino Acid Sequence↗

The Caenorhabditis elegans genes sqv-2 and sqv-6, which are required for vulval morphogenesis, encode glycosaminoglycan galactosyltransferase II and xylosyltransferase.

In mutants defective in any of eight Caenorhabditis elegans sqv (squashed vulva) genes, the vulval extracellular space fails to expand during vulval morphogenesis. Strong sqv mutations result in maternal-effect lethality, caused in part by the failure of the progeny of homozygous mutants to initiate cytokinesis and associated with the failure to form an extracellular space between the egg and the eggshell. Recent studies have implicated glycosaminoglycans in these processes. Here we report the cloning and characterization of sqv-2 and sqv-6. sqv-6 encodes a protein similar to human xylosyltransferases. Transfection of sqv-6 restored xylosyltransferase activity to and rescued the glycosaminoglycan biosynthesis defect of a xylosyltransferase mutant hamster cell line. sqv-2 encodes a protein similar to human galactosyltransferase II. A recombinant SQV-2 fusion protein had galactosyltransferase II activity with substrate specificity similar to that of human galactosyltransferase II. We conclude that C. elegans SQV-6 and SQV-2 likely act in concert with other SQV proteins to catalyze the stepwise formation of the proteoglycan core protein linkage tetrasaccharide GlcAbeta1,3Galbeta1, 3Galbeta1,4Xylbeta-O-(Ser), which is common to the two major types of glycosaminoglycans in vertebrates, chondroitin and heparan sulfate. Our results strongly support a model in which C. elegans vulval morphogenesis and zygotic cytokinesis depend on the expression of glycosaminoglycans.

Amino Acid Sequence↗

Cloning and expression analysis of two mucin-like genes encoding microfilarial sheath surface proteins of the parasitic nematodes Brugia and Litomosoides.

In several filarial genera the first stage larvae (microfilariae) are enclosed by an eggshell-derived sheath that provides a major interface between the parasite and the host immune system. Analysis of the polypeptide constituents of the microfilarial sheath from the cotton rat filaria Litomosoides sigmodontis identified two abundant surface glycoproteins: Shp3a and Shp3. The corresponding genes and the orthologues of the human parasite Brugia malayi and the rodent filaria Brugia pahangi were cloned and sequenced. They encode secreted, mucin-like proteins with N-terminal Ser/Thr-rich repeats and a C-terminal anchor domain rich in aromatic amino acids. About 75% of the protein molecular masses result from post-translational modifications. The Ser/Thr-rich motifs are supposed to serve as targets for dimethylaminoethanol-phosphate substitutions. These modifications were detected only on the sheaths of the late developmental stage of stretched microfilariae, corresponding with the expression of the proteins in the epithelium of the distal part of the uterus and the specific transcription of shp3 and shp3a in the anterior female worm segment. Genomic analysis of all three species demonstrated a conserved linkage of the two genes. Their transcripts undergo cis- and trans-splicing. The transcription start sites of the primary transcripts were determined for the L. sigmodontis genes. The core promoter regions are remarkably conserved between the paralogue genes Ls-shp3a and Ls-shp3 and their orthologues in Brugia, implicating conserved regulatory elements.

Amino Acid Motifs↗

Characterization of a novel filarial serine protease inhibitor, Ov-SPI-1, from Onchocerca volvulus, with potential multifunctional roles during development of the parasite.

A novel filarial serine protease inhibitor (SPI) from the human parasitic nematode Onchocerca volvulus, Ov-SPI-1, was identified through the analysis of a molting third-stage larvae expressed sequence tag dataset. Subsequent analysis of the expressed sequence tag datasets of O. volvulus and other filariae identified four other members of this family. These proteins are related to the low molecular weight SPIs originally isolated from Ascaris suum where they are believed to protect the parasite from host intestinal proteases. The two Ov-spi transcripts are up-regulated in the molting larvae and adult stages of the development of the parasite. Recombinant Ov-SPI-1 is an active inhibitor of serine proteases, specifically elastase, chymotrypsin, and cathepsin G. Immunolocalization of the Ov-SPI proteins demonstrates that the endogenous proteins are localized to the basal layer of the cuticle of third-stage, molting third-stage, and fourth-stage larvae, the body channels and multivesicular bodies of third-stage larvae and the processed material found between the two cuticles during molting. In O. volvulus adult worms the Ov-SPI proteins are localized to the sperm and to eggshells surrounding the developing embryos. RNA interference targeting the Ov-spi genes resulted in the specific knockdown of the transcript levels of both Ov-spi-1 and Ov-spi-2, a loss of native proteins, and a significant reduction in both molting and viability of third-stage larvae. We suggest the Ov-SPI proteins play a vital role in nematode molting by controlling the activity of an endogenous serine protease(s). The localization data in adults also indicate that these inhibitors may be involved in other processes such as embryogenesis and spermatogenesis.

Amino Acid Sequence↗

Comparison of premorbid and postinjury mmpi-2 profiles in late postconcussion claimants.

A comparison of premorbid and postinjury MMPI-2 profiles was performed in 23 patients with mild cranial/cervical injuries. All claimants attributed major personality change to their injuries during the course of compensation-related neuropsychological examinations. Their premorbid MMPI-2 profiles were all abnormal and the modal code-type indicated somatoform psychopathology. The post-accident MMPI profiles showed continuous somatization trends, but they unexpectedly showed (a) increased defensiveness and (b) a general decrease in global psychopathology. The findings did not support an 'eggshell plaintiff' theory of chronic postconcussive complaints. The view that chronic postconcussive complaints require a wider focus on non-neuropsychological factors is enhanced.

Accidents, Traffic↗

Effects of dietary L-carnitine on the performance and egg quality of laying hens from 65-73 weeks of age.

The effects of L-carnitine supplementation (50-500 mg/kg diet) of a practical layer diet, based on maize, soyabean and wheat, on the performance of laying hens and some indices of egg quality were studied for 8 weeks, using 65-week-old hens kept in cages. Albumen quality (albumen height and Haugh (1937) unit score) was improved, while yolk index and yolk colour score were not affected by dietary L-carnitine. The percentage of egg-white increased and that of egg yolk decreased in response to dietary supplementation of L-carnitine. Dietary L-carnitine did not influence laying performance (egg production rate, mean egg weight, daily feed intake, daily egg mas and feed conversion) or external egg quality measured by egg weight, egg-shape index or by eggshell quality, either measured directly as shell breaking strength or indirectly as shell weight, shell thickness or shell weight per unit surface area. Based on the results of the present study, L-carnitine had a beneficial effect on albumen quality and could modify the components of the edible part of the egg, during the late laying period.

Animals↗

Intestinal function and reproductive capacity of Tegel pullets in response to exogenous oestrogen.

The effects of varying levels of exogenous oestrogen (E2) (0, 10 or 100 micrograms E2/kg BW) on the development of 18-week old pullets were tested over a 28-day period. The hormone had no significant effects on feed intake, body growth, feed conversion ratio or weight of the oviduct. Similarly, there were no significant effects of the hormone on egg production and egg weight but eggshell thickness and weight of shell per unit area were increased (P < 0.05) at a lower level of administration (10 micrograms E2/kg BW), compared to the control and the highest level of hormone. The morphometry of the jejunal mucosa and some enzymes associated with Ca transport were similar between the three groups. Oestrogen treatment, however, intensely enhanced the expression of calbindin D22K, although this was not quantified.

Animals↗

Effects of low dietary protein and different allocations of food during rearing and restricted feeding after peak rate of lay on egg production, fertility and hatchability in female broiler breeders.

1. A 3x2x2 factorial experiment was conducted with boiler breeder females to determine the effects of body weight gain (three levels) and conventional or lower crude protein concentrations in the rations during rearing, and restricted or ad libitum feeding after the peak rate of lay on egg production, fertility, hatchability, mortality and food consumption. 2. Compared with ad libitum feeding, conventional food restriction resulted in a decrease in average daily food consumption of 0.6 during rearing, 0.2 during early lay and an increase of 0.4 after the peak rate of egg production. Mortality was decreased by more than half. 3. Restricted birds had higher total and settable egg production, fewer defective or damaged eggshells and higher fertility and hatchability than those fed ad libitum. The modified (more generous) rearing programme resulted in lower rates of egg production and higher rates of mortality compared with the conventional food restriction programme. 4. Low-protein rearing rations were associated with higher rates of food intake, higher mortalities and lower rates of egg production than the conventional protein rations. There were no differences in the fertility or hatchability of eggs between birds fed on the two concentrations of dietary crude protein. 5. Ad libitum feeding post-peak was associated with higher rates of mortality to 60 weeks of age. Post-peak feeding had little effect on the rate of lay or egg weight in conventionally restricted birds fed high protein rations or in birds fed ad libitum. Restricted feeding post-peak decreased the rate of lay and egg weight in birds on the modified restriction programme and in conventionally restricted birds fed on the low protein rations. There was no effect of post-peak feeding on fertility or hatchability of eggs. 6. The total numbers of saleable chicks per kg food consumed were 1.83, 1.72 and 0.52 for conventional, modified and ad libitum feeding during rearing; 1.56 and 1.15 for restricted and ad libitum feeding post-peak: there were no differences associated with protein concentrations of rations fed during rearing.

Animal Welfare↗