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Calcium-activated ATPase of the chick embryonic chorioallantoic membrane. Identification, developmental expression, and topographic relationship with calcium-binding protein.

A Ca2+-activated ATPase activity is present in the chick embryonic chorioallantoic membrane (CAM), the placenta-like tissue which translocates eggshell calcium into the embryonic circulation. The enzyme is membrane-bound, ATP-specific, Mg2+-dependent, exhibits dual Km values of 30 microM and 0.3 mM Ca2+, and has a Mr of 170,000. Throughout embryonic development, a single electrophoretic form of the Ca2+-ATPase is found and, furthermore, its specific activity as a function of age follows a bimodal pattern. In particular, from incubation days 14-15 to the end of gestation, a period representing rapid embryonic calcium accumulation, Ca2+-ATPase specific activity increases 6-fold. Cytohistochemistry localized the Ca2+-ATPase exclusively within the CAM ectoderm which lies adjacent to the calcium-rich shell membrane/eggshell. In a parallel study, cleavable bifunctional cross-linking agents were used to characterize the in situ protein topography of the CAM ectodermal surface adjacent to the calcium-binding protein (CaBP), a CAM cell-surface protein associated with calcium transport. We found that the immediate near neighbor of the CaBP is a 170,000 Mr, membrane-bound protein. The 170,000 protein was co-isolated with the CaBP after cross-linkage in situ and subsequent immunoprecipitation with anti-CaBP antibodies. Reductive cleavage of the immune complex released detectable Ca2+-ATPase activity, suggesting that the 170,000 protein is the Ca2+-ATPase of the CAM.

Allantois↗

Silkmoth chorion proteins. Their diversity, amino acid composition, and the NH-terminal sequence of one component.

Silkmoth eggshell (chorion) proteins have been characterized by electrophoresis on sodium dodecyl sulfate and isoelectric focusing polyacrylamide gels; up to 33 and 41 components, respectively, were detected from a single chorion. Some of these components are polymorphic, being absent from chorions of certain animals. A system of nomenclature for all chorion proteins is presented, based on their separation on sodium dodecyl sulfate and isoelectric focusing gels. The chorion is enriched in glycine, alanine, cysteine, and tyrosine and poor in methionine and histidine. The proteins were fractionated into four partially overlapping groups; all four are enriched in the above amino acids, although significant differences exist. Further fractionation by isoelectric focusing of one of the above groups, s/s, yielded seven components, two of which are homogeneous both on sodium dodecyl sulfate and isoelectric focusing gels. The amino acid compositions, molecular weights, and solubility properties of the components share certain features which distinguish s/s as a group from the other three groups. The sequence of the first 67 NH2-terminal residues of a homogeneous protein purified from s/s has been determined. The protein contains a cysteine-rich tail (3 cysteines in the first 18 residues) followed by a 49-residue segment which contains only a single cysteine residue. This latter segment also contains two different tetrapeptide sequences which are each repeated, one twice and the other four times.

Amino Acid Sequence↗

Lagochilascaris sprenti sp. n. (Nematoda: ascarididae) from the opossum, Didelphis virginiana (Marsupialia: didelphidae).

Sixty-eight specimens of Lagochilascaris from the stomach of seven naturally-infected opossums from Louisiana were described as Lagochilascaris sprenti sp. n. Observations were also made on adult worms of both sexes recovered from experimentally-infected opossums and mice. The new species differs from the four previously described species of Lagochilascaris in the length of the spicules, number of pits on the circumference of the egg, and position of the vulva. The spicules are shorter in L. sprenti than in L. turgida and L. buckleyi. The number of surface pits in the eggshell at the circumference of the egg is 24 to 31 in L. sprenti, less in L. minor (15-26), and more in L. major (33-45). In worms of equal length, the vulva is consistently about 1 mm further anteriad in L. sprenti than it is in L. minor or L. major.

Animals↗

Staphylococcal food poisoning associated with an Easter egg hunt.

Staphylococcal contamination of intact, hard-boiled eggs resulted in the food poisoning of an estimated 300 children out of 850 who had participated in an Easter egg hunt. Enterotoxigenic staphylococci that were isolated from the Easter eggs matched that obtained from an infected cook who prepared the eggs three to five days before the hunt and which he left unrefrigerated. Experimental studies demonstrated that heated eggs can absorb 2 mL of contaminated cool water through intact eggshells. When water was inoculated with pathogenic staphylococci at even low contamination levels, rapid growth and enterotoxin production within cooked eggs could be easily duplicated. This is the first large outbreak of its type; safeguards can and should be employed to prevent future ones.

Adolescent↗

Organochlorine residues and shell thicknesses in eggs of the clapper rail, common gallinule, purple gallinule, and limpkin (class Aves), eastern and southern United States, 1972-74.

Organochlorine residues and shell thicknesses were surveyed in eggs of the clapper rail (Rallus longirostris), purple gallinule (Porphyrula martinica), common gallinule (Gallinula chloropas), and limpkin (Aramus guarauna) from the eastern and southern United States. Clapper rail eggs were collected during 1972-73 in New Jersey, Virginia, and South Carolina. During 1973-74, gallinule eggs were collected in Florida, South Carolina, and Louisiana, and limpkin eggs were collected in Florida. Egg contents were analyzed for residues of organochlorine pesticides, including DDT, TDE, DDE, dieldrin, mirex, heptachlor epoxide, oxychlordane, cis-chlordane (and/or trans-nonachlor), cis-nonachlor, hexachlorobenzene (HCB), toxaphene, and endrin, and for polychlorinated biphenyls (PCBs). Shell thicknesses of recent eggs of these species were compared with archival eggs that had been collected before 1947. With the exception of the limpkin, the majority of eggs analyzed contained residues of p,p'-DDE and PCBs. Geometric means ranged from 0.10 ppm to 1.3 ppm. Small amounts (less than 1.0 ppm) of mirex, dieldrin, cis-chlordane (and/or trans-nonachlor), TDE, and DDT were detected in a few eggs. No evidence of eggshell thinning was found for any of the species studied. DDE residues in clapper rail eggs were higher in New Jersey and Virginia than in South Carolina.

Animals↗

Review: the value of the American alligator (Alligator mississippiensis) as a model for research in craniofacial development.

American alligators exhibit a number of morphological features which are more characteristic of mammals than reptiles. This unique combination makes an extremely useful animal for experimental investigations of craniofacial development. Thus, it is possible to make a longitudinal study of the development of the alligator's secondary palate by utilizing shell-less culture techniques. Incubation of eggs at 28 degrees C, as opposed to 3 degrees C, slows development sufficiently to enable both the recovery of critical intermediate stages, and also great precision in the timing of teratogen administration. Injection of either 0.01 mg of 5-fluoro-2-desoxyuridine or 0.05 mg of hydrocortisone into the yolks of Days 12, 14, and 17 embryos produces cleft lip, cleft lip and palate, and cleft palate, respectively. Application of the teratogen at critical developmental stages on Day 12 reliably induces unilateral cleft lip on either the right or left sides, while injection on Day 10 yields embryos with normal lips and palates but virtually no lower jaw. The spontaneous malformation rate is very low, but highest in eggs laid by young and old animals. Changes in eggshell banding provide a noninvasive method for monitoring embryonic development. Detailed comparisons of palatal development in birds and alligators permit an accurate phylogenetic study of physiological clefting in the aves. The alligator dentition is readily amenable to experimental studies.

Abnormalities, Drug-Induced↗

Chemical synthesis, biological activity, and metabolism of 25-hydroxy-24-oxovitamin D3.

25-Hydroxy-24-oxovitamin D3 (25(OH)24-oxo-D3), a metabolite of 25-hydroxyvitamin D3, has been chemically synthesized. The ultraviolet, mass, infrared, and proton nuclear magnetic resonance spectra of the 25(OH)24-oxo-D3 were identical with those of the natural product isolated from chick kidney incubates. The oxo compound showed biological activity similar to 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) in vitamin D-deficient chicks in enhancing intestinal calcium transport and bone calcium mobilization activities. Although 25(OH)24-oxo-D3 partially restored the impaired eggshell weights of Japanese quails fed a vitamin D-deficient diet, it was much less potent than 25-hydroxyvitamin D3 or 1 alpha, 25-dihydroxyvitamin D3. In addition, there was no effect on the calcification of medullary bone. When 25(OH)24-oxo[3H]D3 was incubated with kidney homogenates from vitamin D-deficient chicks, it was metabolized to [3H]1 alpha, 24,25-trihydroxyvitamin D3 and a metabolite which was eluted in a region between authentic 24R,25(OH)2D3 and 1 alpha, 25-dihydroxyvitamin D3 on high pressure liquid chromatography. In the incubates of kidney homogenates from vitamin D-supplemented chicks, those metabolites were not detected. In vitamin D-supplemented chicks, the recovery of radioactivity in the chloroform phase extracted by the method of Bligh and Dyer was only 50%, while that in vitamin D-deficient chicks was 87%. Moreover, the radioactivity eluted in the 25(OH)24-oxo-D3 fraction from vitamin D-supplemented chicks was only one-fifth of that from vitamin D-deficient birds. The present results indicate that the 24-oxidation of 24,25(OH)2D3 may be a route of inactivation of vitamin D3.

Animals↗

Organochlorine and heavy metal residues in black duck eggs from the Atlantic Flyway, 1978.

Black duck (Anas rubripes) eggs were collected during 1978 in the Atlantic Flyway. One egg from each of 49 clutches was analyzed for organochlorine compounds and mercury. DDE was detected in 39 eggs, ranging from 0.09 ppm to 3.4 ppm, wet weight. DDE residues were highest in eggs from Delaware, where the mean DDE level was 2.0 ppm. DDT and TDE were present at low levels in only five and four eggs, respectively. PCBs resembling Aroclor 1260 were detected in 24 eggs and ranged from 0.43 ppm to 2.9 ppm. Eggs from Massachusetts and Rhode Island contained an average of greater than 1.0 ppm PCBs, but eggs from Nova Scotia, Pennsylvania, Maryland, and Virginia contained no detectable PCBs. Dieldrin, oxychlordane, and heptachlor epoxide were present in a few samples at low levels. Mercury was detected in 31 eggs, ranging from 0.07 ppm to 0.34 ppm, wet weight. Twenty eggs analyzed for chromium, copper, and arsenic contained averages of 0.64 ppm, 1.7 ppm, and 0.18 ppm, respectively. No geographic pattern was observed in these metal residue levels. Eggshell thickness (0.347 mm) was identical to the pre-1946 norm.

Animals↗

Calcium homeostasis in in ovo and ex ovo turkey embryos.

Calcium homeostasis of in ovo (normal) and ex ovo (shell-less) turkey embryos was investigated at 15, 18, and 21 days of incubation. Hypocalcemia and an elevation in circulating 1,25(OH)2D3 in ex ovo embryos were observed by 15 days. Calcium and phosphorus concentrations in femora and tibiae in ex ovo embryos were significantly lower compared to their normal counterparts. These results suggest that shell calcium mobilization is required prior to 15 days of incubation for maintaining serum calcium and supporting bone mineralization. Furthermore, the elevation of 1,25(OH)2D3 is indicative of a functional calcium homeostatic mechanism responding to the absence of the primary calcium source (eggshell) during the second half of turkey embryonic development.

Animals↗

Current concepts and management of patients with fixed decompensated spinal deformity.

The patient with a fixed decompensated spine deformity presents a difficult orthopaedic challenge. Consideration of the potential risks and benefits is critical with these patients before embarking on a significant surgical procedure. The deformity should be evaluated according to coronal, sagittal, and axial planes, with consideration given to where the major deformity exists. For deformities which exist in the sagittal plane only, a single stage posterior decancellation, known as the eggshell procedure, and posterior spinal fusion are preferred, rather than a 2 stage combined anterior and posterior approach. For deformities in the coronal plane, which are often associated with sagittal and axial malalignment, combined approaches are preferable for correcting imbalance. However, for severe, multiplane deformities where multiple osteotomies and recorrection are unlikely to safely produce balance, vertebral body resection with spinal shortening would be preferable. The authors' experience in treating fixed, decompensated spinal deformities is presented herein.

Adolescent↗

Organochlorine residues in Alaskan peregrines.

Organochlorine residue levels in eggs of Alaskan peregrines have remained essentially constant over the period 1969-73 despite decreased usage of these compounds in the United States and Canada. Studies on reproductive success in Great Britain and data on eggshell-thinning suggest that DDE residues above 20 ppm wet weight in peregrine eggs are associated with inability to maintain population levels. Residues in mainland Alaska are well above this critical figure and the reproductive rate is low. On the Colville River in northwestern Alaska, the last young falcons will fledge in 1975 and the remaining adult population will disappear by 1980 unless the present rate of reproductive failure is drastically and quickly reversed. In the Aleutians, however, levels range from 5 to 7 ppm and the reproductive rate is adequate to maintain the population.

Alaska↗

The influence of Datura ferox alkaloids on egg-laying hens.

Seeds of the weed Datura ferox are frequent contaminants of raw materials used for animal feed. In this study a mixture of scopolamine and hyoscyamine (98:2), the 2 main alkaloids of Datura ferox seeds, was incorporated at 4 total alkaloid levels (1.5, 15, 75 or 150 mg/kg feed) into a control diet fed to 100 egg-laying hens for 3 mo. Alkaloid doses of 150 mg/kg feed reduced egg production for the first 5-6 w of feeding, whereas lower doses had no effect. Egg weight, eggshell thickness and body weight of hens were unaffected at all doses. Doses of 150 mg/kg feed produced significant increases in the cardiac rate of hens after 5 w. Breathing frequency at all doses was unaffected. Determination of plasma aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and leucine aminopeptidase activities, as well as autopsy and histological examinations, revealed no obvious alkaloid-related toxic effects. It was concluded that a total alkaloid dose as high as 75 mg/kg feed can be safely administered to egg-laying hens.

Animal Feed↗

Fatty liver-hemorrhagic syndrome observed in commercial layers fed diets containing chelated minerals.

This study was originally conducted to determine the effect of various chloride levels and consumption of chelated versus non-chelated minerals on egg production and eggshell breaking strength. However, the focus of this report changed after fatty liver-hemorrhagic syndrome (FLHS) was observed in hens that consumed diets containing chelated minerals. No FLHS was observed in hens that consumed a diet that contained non-chelated minerals. Four hundred thirty-two 35-week-old commercial laying hens were housed in individual cages in groups of nine hens each. The treatments were factorially arranged (3 x 2) such that six experimental diets differing in chloride levels and in source of minerals were fed for six 28-day laying periods. A significantly higher mortality attributed to FLHS was observed in hens that consumed the diets having chelated minerals as compared with hens that consumed the diets with non-chelated minerals. No difference in mortality was observed among the hens fed the various chloride diets.

Animal Feed↗

Effect of Eimeria tenella infection on the production of Salmonella enteritidis-contaminated eggs and susceptibility of laying hens to S. enteritidis infection.

Experiments were conducted to determine the effect of Eimeria tenella infection on the production of Salmonella enteritidis-contaminated eggs and susceptibility of egg-laying hens to S. enteritidis infection. The dose of sporulated oocysts of E. tenella used in the present study was 2 x 10(5) per chicken. When hens were infected with 10(6) colony-forming units (CFU) of S. enteritidis, E. tenella infection resulted in a significant increase (P < 0.05) of S. enteritidis contamination in the shells of fresh eggs but not in the egg contents of fresh eggs and stored eggs. Significant effects of coccidial infection on the contamination of eggs (shells and contents) were not observed when hens were infected with 10(4) CFU of S. enteritidis. Surface-disinfected eggs pooled within each week from individual chickens infected with 10(4) CFU of S. enteritidis and E. tenella showed significantly (P < 0.05) more S. enteritidis contamination in eggshells, but not in egg contents, than did those from hens infected with S. enteritidis alone. Coccidial infection resulted in a significant increase of S. enteritidis in the cloacal swabs of hens infected with 10(4) CFU of S. enteritidis. In all the experiments, chickens infected with S. enteritidis and E. tenella showed significantly higher positive rates and counts of cecal S. enteritidis than did chickens infected with S. enteritidis alone. No significant differences of S. enteritidis-positive rates in ovary, oviduct, liver, and spleen were noted whether the hens were infected with a combination of E. tenella and S. enteritidis or with S. enteritidis alone.

Animals↗

Organochlorine insecticide residues in African Fauna: 1971-1995.

Organochlorine insecticides (OCLs), which were introduced in the decade following World War II, were used extensively in Europe, the U.S., and other developed countries into the 1970s. However, data began to accumulate on their persistence in soils and aquatic sediments, their potential to be taken up into animal tissues and to bioconcentrate in birds and mammals in the higher tropic levels of food chains and even in humans. As a result, registration authorities phased out their use progressively, in Europe and the U.S., from 1973 onward. However, the production of OCLs in developed countries and their use in developing countries continued through the 1970s and 1980s into the 1990s because they were, no longer under patent agreement, were inexpensive to manufacture, and were very effective in pest control. In Africa, the use of OCLs continued well into the 1990s for the control of mosquitoes, tsetse flies, and desert locusts as well as to combat various crop, animal, and human pests. Some of these uses involved extensive spraying of large areas of nonagricultural land, thereby exposing many groups and species of wildlife to their residues. Although there is some evidence of a gradual decline in the use of OCLs in Africa, they are still being used in appreciable quantities. During the past 25 yr, there have been 50 published reports of OCL residues in the various groups of invertebrate and vertebrate animals constituting the African fauna. These have been based on a diverse range of surveys, target animals, sampling methods, and analytical techniques. Moreover, they are extremely regionally-biased, the most intense surveys being in Zimbabwe, Kenya, Egypt, and South Africa. DDT was the most commonly used OCL, accounting for about half the total use, followed closely by dieldrin and HCH. Birds and fish have been sampled most intensively, with relatively few studies on other taxa. We reviewed the OCL residue data on African fauna from these reports and summarized the maximum and mean residues in the various groups of terrestrial and aquatic invertebrates and vertebrates. Overall, residues in the fauna were the greatest for DDT, followed in turn by those of dieldrin, HCH, endosulfan, and endrin, with small amounts of aldrin and toxaphene being found in some animals. There were relatively few reports of OCL residues in terrestrial invertebrates and virtually none in aquatic invertebrates. Only a few reports demonstrated OCL residues in terrestrial vertebrates, although high levels of DDT, dieldrin, and HCH were found in crocodile eggs and large residues of dieldrin occurred in bats, squirrels, and monkeys. Considerable OCL residues were reported in a few species of fish, especially Barbus, Clarias, Hydrocynus, Labeo, Sarotherodon, Epiplatys, and Synodontis. These residues were at levels that could have caused chronic toxicity or behavioral changes. The calculated maximum and mean OCL residues in the various elements of the African fauna until 1995 were compared with those calculated for corresponding faunal groups in Europe and the U.S. from their development and introduction up to 1973. The OCL residues reported in African fauna between 1971 and 1975 tended to be significantly higher overall than those published for Europe and the U.S. In particular, residues of DDT and dieldrin in African birds and their eggs were greater than those that had been incriminated as causing significant eggshell thinning and reproductive failure in European and U.S. aquatic and terrestrial birds up to 1973. Additionally, high DDT and dieldrin residues were reported from some species of African fish at levels that could potentially affect their reproduction, have chronic toxic and behavioral effects, and even drastically affect populations. Holistic case studies on the use of OCLs to control tsetse flies and desert locusts were discussed. OCL levels in trophic levels of fauna associated with Lake Kariba (between Zambia and Zimbabwe) were summarized. (AB

Africa↗

Assessment of temporomandibular injury and orofacial pain.

Every individual has physical faults which makes him less than a perfect biological specimen. This is the foundation for the legal "eggshell principal." Most of us do not have perfect skeletal formation, muscular adaptation, neurological or vascular systems. Additionally, we may not have complete symmetry and ideal form to our cranium or mandible. Our dentitions are imperfect. All of these imperfections play a role in every injury. The variances of normal that are seen in any medical or dental population, must be accepted; therefore, a given injury can have tremendously varied sequelae, depending on the person receiving it. The patho-physiology resultant from an accident may not parallel the severity of the motor vehicle accident, fall or blow. Prompt and proper evaluation of the individual-turned-patient is imperative. Referral to a dentist, competent in providing the necessary evaluation of the injury, is the responsibility of those individuals attending to the injured person's physical, emotional, and financial trauma. Their physician, dentist, attorney or counselor should take the responsibility of providing proper referral. The four most dangerous words in medicine and dentistry today are: "It Will Go Away." People often develop serious sequelae by assuming that their problem will "go away" by just leaving it alone long enough or by simply ignoring it. With proper evaluation and management, most trauma related temporomandibular injuries can be corrected or at least be properly managed.

Facial Pain↗

p,p'-DDE, polychlorinated biphenyls, and endrin in oldsquaws in North America, 1969-73.

Organochlorinated compounds were monitored in oldsquaws (Clangula hyemalis) and their food from Lake Michigan between October and May, 1969-72; in adult oldsquaws, eggs, young, and food from northwest Hudson Bay in 1971; and in oldquaws from five wintering areas other than Lake Michigan in 1971-73. Analyses were conducted on 300 carcasses, 14 wings, 29 gullet samples, and 11 clutches. Average residues in carcasses from Lake Michigan ranged from 4 to 107 ppm PCBs, 2 to 42 ppm DDE, and less than 0.1 to 0.7 ppm endrin. Differences in DDE levels occurred between several sex and age classes during December on Lake Michigan; these differences were not apparent in the spring. Increases in DDE and PCB residues for oldsquaws occurred on Lake Michigan between December and May. DDE residues in the wing and carcass were significantly correlated. Residues were relatively low in oldsquaw foods from Lake Michigan; concentration factors between the food and the ducks varied between 1X and 22X, depending on the date and compound. Organochlorinated residues were lower in Arctic than in Lake Michigan food samples. DDE in paired male and female oldsquaws was highly correlated, as was DDE in females and clutches. Eggshell thickness had declined 4.5% compared with eggs collected before 1947. Residues were higher in oldsquaws wintering on the Great Lakes and lowest in oldsquaws from coastal areas.

Age Factors↗