Collagenaceous,thiol-containing proteins of cnidarian nematocysts: A comparison of the chemistry and protein distribution patterns in two types of cnidae

• 1.1. Nematocyst structural proteins (NSP) from the sea anemones Aiptasia pallida and Metridium senile and the siphonophore Physalia physalis are primarily low molecular weight collagens linked by disulfide bonds.
• 2.2. NSP patterns resolved by SDS-PAGE revealed a common, major collagen species (40 kDa) in each nematocyst type, together with other collagens and non-thiol-containing proteins.
• 3.3. For each cnidarian, NSP glycosylation profiles were significantly different.
• 4.4. Monoclonal antibodies against Aiptasia NSP demonstrated a differential distribution between capsule wall and thread.
• 5.5. NSP differences would account for the diversity of morphologic and functional types.

     

Cloning and characterization of rabbit pancreatic colipase

• 1.1. Among the digestive enzymes synthesized by pancreas, lipase is the principle lipolytic enzyme which hydrolyses dietary glycerides.
• 2.2. For its action it requires a coenzyme, colipase.
• 3.3. The molecular mechanisms of the interaction of these two are not fully understood.
• 4.4. Further, molecular events that regulate and influence lipid absorption are ill denned.
• 5.5. The rabbit is the conventional animal model for the study of lipid absorption. We have undertaken the molecular cloning, and characterization of rabbit pancreatic colipase, the coenzyme for pancreatic lipase.
• 6.6. Colipase has been cloned from a gt 11 library of an adult rabbit pancreatic cDNA by probing with an oligonucleotide derived from human colipase sequence.
• 7.7. The total reading frame consists of 321 nucleotides coding for 90 amino acids of the functional protein and 17 nucleotides of the leader peptide.
• 8.8. Northern blot analysis revealed a distinct band around 0.5kb. Comparison with other species revealed an over all homology of 75% at the nucleotide level.
• 9.9. At the amino acid level highest conservation is observed at the lipase-binding region (AA 53–73).
• 10.10. Rabbit enzyme also retained the N-terminal pentapeptide of it preform.
• 11.11. The regions of homology and conservation may aid to define the sites of interaction of colipase with lipase.

     

Accumulation,distribution and toxicity of selenium in the adult house fly,Musca domestica

• 1.1. Accumulation and distribution of dietary Se in relation to mortality was investigated in adult house flies.
• 2.2. The midgut preferentially accumulated Se and thereby limited toxicity.
• 3.3. Midgut Se concentrations were from 6- to 107-fold higher than in carcass, and from 15 to 71% of the total Se was associated with midgut.
• 4.4. When dietary levels of Se were raised the midgut saturated at 15 μg Se/g tissue, followed by a rise in carcass levels to greater than 0.5 μg Se/g tissue and increased mortality.
• 5.5. Se levels in lysosomal fractions were from 3- to 50-fold higher than in other subcellular fractions, suggesting that Se is sequestered in lysosomes.
• 6.6. Se added to drinking water was toxic at 4–8 ppm.

     

Collagen diversity in the sea urchin,strongylocentrotus purpuratus

• 1.1. Pepsin insensitive fragments of collalgen extracted from tube feet and peristome have α 1 and α 2 bands that differ in apparent molecular weights from each other and from human type 1 collagen.
• 2.2. A monoclonal antibody that reacts with the ξ I band and a low molecular weight fragment of tube foot collagen does not react with either peristome collagen or human type I collagen.
• 3.3. Measurements of the axial periodicity of native fibers of tube foot and peristone collagens indicate they have D values that differ significantly from each other and from reported values of vertebrate type I collagen.
• 4.4. We propose that there are diverse and specialized types of collagen in sea urchins that are heterogeneously distributed in the extracellular matrix of different tissue.

     

The major serum protein of fetal and newborn pigs: Biochemical properties and identification as a fetal form of α1-acid glycoprotein

• 1.I. The major protein component of fetal pig serum, has been immunologically identified as α₁-acid glycoprotein (orosomucoid).
• 2.2. Amino acid composition and total carbohydrate content (around 38% by weight) were similar in the adult and fetal forms of α₁-acid glycoprotein. These forms differ, however, in the proportion of individual monosaccharides.
• 3.3. Fucose, represented the 1.5% (by weight) in the fetal protein, and the 2.5% in its adult counterpart. The latter was more susceptible to ncuraminidase and also possesses a higher mannose/galactose ratio than the fetal form.
• 4.4. Insolubilized Concanavalin A (Con A) retained 80%, of the adult protein, whereas the fetal form was mostly Con A-non reactive. The proportion of this -non reactive fraction, as revealed by crossed immuno-affino-electrophoresis experiments, was age-dependent and varied from 62% at fetal age of 50–60 days to 80% at birth.

     

Heart rates of adult red-spotted newts,Notophthalmus viridescens in air and submerged in normoxic and hypoxic water at different temperatures

• 1.1. Heart rates of adult aquatic red-spotted newts can be conveniently recorded using an impedance pneumograph.
• 2.2. Heart rates decrease linearly with decreasing temperature.
• 3.3. Submergence in normoxic and hypoxic water at 10°, 15°, and 20°C results in bradycardia which is more pronounced in hypoxic water.
• 4.4. At 5°C one newt exhibited the above pattern, but bradycardia was not exhibited by the other newt during normoxic submergence.
• 5.5. Diminishing heart rates are probably due to oxygen deficiency, not immersion alone.
• 6.6. Recovery from bradycardia in air is rapid and not linked with resumption of aerial breathing.

     

Acetylcholinesterase in Apis mellifera head during post-embryonic development. Existence of a glycoinositol-anchored membrane form at eary pupal stages

• 1.1. Properties of acetylcholinesterase (AChE, EC 3.1.1.7) from Apis mellifera head were studied during pupal development and at the adult stage.
• 2.2. During post-embryonic development, tissue and specific activities were closely related and increased to reach a maximum value at emergence and at last pupal stage, respectively.
• 3.3. In adults, AChE activity was weaker in foragers than in emerging bees.
• 4.4. The membrane form occurred in adult bees as well as in pupae whereas the soluble enzyme only appeared from Pd pupal stage.
• 5.5. The proportion of soluble and membrane forms fluctuated during late development but, in all cases, the percentage of the soluble form remained less than 10% of total AChE activity.
• 6.6. At all post-embryonic stages, the membrane form was sensitive to the action of phosphatidylinositol-specific phospholipase C (PI-PLC) and was converted into a hydrophilic enzyme.
• 7.7. In adult bees, the sensitivity to PI-PLC depended on the season. In summer, about 60% of the membrane activity could be solubilized by PI-PLC vs only 5% in winter.
• 8.8. The sensitivity of AChE to pirimicarb varied with the developmental stage.
• 9.9. In foraging bees, AChE was more susceptible to pirimicarb than in emerging bees. This difference of sensitivity to carbamate was abolished after removal of the membrane anchor either by mild trypsin digestion of PI-PLC treatment.

     

A fetal-maternal shift in the oxygen equilibrium of hemoglobin from the viviparous caecilian,Typhlonectes compressicauda

• 1. The equilibria and kinetics of oxygen binding by blood and hemoglobin from adult and fetal caecilians,Typhlonectes compressicauda, have been measured.
• 2. The oxygen affinity of fetal blood is higher than that of adult blood.
• 3. Electrophoresis of adult and fetal hemoglobins suggests that they may be identical: a major and minor component occurs in each.
• 4. Adult and fetal hemoglobins have identical oxygen equilibria. Stripped hemoglobins have a high oxygen affinity and no Bohr effect between pH 6.5 and 10.0. An “acid”, reversed Bohr effect is present below pH 6.5. The addition of 1 mM ATP reduces the oxygen affinity markedly and produces a moderate, normal Bohr effect.
• 5. The major nucleoside triphosphate in fetal and adult erythrocytes is adenosine triphosphate: about 10% of the nucleoside triphosphates is guanosine triphosphate. Adult erythrocytes contain 3 times as much ATP as do the fetal erythrocytes.
• 6. The fetal to maternal shift in the oxygen equilibrium is mediated entirely by the difference in ATP content of the maternal and fetal red blood cells.

     

In vitro Ca deposition by rat matrix vesicles: Is the membrane association of alkaline phosphatase essential for matrix vesicle-mediated calcium deposition?

• 1.1. Phosphatidylinositol phospholipase C (PI-PLC) treatment of rachitic rat matrix vesicles (MVs) released about 80% of membrane-bound alkaline phosphatase (ALP), AMPase, PPiase into the media.
• 2.2. About 20% hydrolytic activity was not released from MV membranes by PI-PLC treatment.
• 3.3. SDS-polyacrylamide gel electrophoresis and Western blot analysis showed only one immunoreactive protein corresponding to the molecular weight of ALP present in the soluble fraction after PI-PLC treatment.
• 4.4. The specific activity of the released ALP was at least 5-fold higher than the residual activity.
• 5.5. After PI-PLC treatment, MVs also demonstrated an 80% reduction of AMP- or βGP-dependent calcium deposition.
• 6.6. The soluble fraction containing 80% of ALP activity was unable to support calcium deposition. The mixing of the soluble and insoluble fractions after PI-PLC treatment failed to fully restore calcium-depositing activity.

     

Mixed function oxidase activity in the harbour seal (Phoca vitulina) from sable is., N.S.

• 1.1. One adult male, eight pups (including two full term foetuses) and nine adult female harbour seals (Phoca vitulina) were analysed for indices of mixed function oxidase (MFO) activity.
• 2.2. MFO activity was present in liver samples, but was at or below detection limits in samples of kidney, lung and pancreas.
• 3.3. Hepatic ethoxyresorufin O-de-ethylase and benzo[a]pyrene hydroxylase activities were similar to those reported in other seals and in other mammals.
• 4.4. Cytochromes P-450 and b₅ concentrations were slightly lower than those observed in other mammals.
• 5.5. MFO activities in newborn pups and foetuses were significantly lower than those in adult females.
• 6.6. No qualitative differences in cytochrome P-450 isozyme distribution between foetal and adult samples could be discerned by electrophoresis.

     

Lipid digestion in cod (gadus morhua)

• 1.1. Cod, 2.6–3.4 kg. were fed a mixed diet of sprat, capelin oil and wheat flour.
• 2.2. Lipids from the feed, stomach and four intestinal segments were separated into tri-, di- and monoglycerides and free fatty acids and analysed by GLC.
• 3.3. All lipolytic products were concentrated in 14:0, 16:0 and 18:0, up to 60% and extremely low in the ω-3 fatty acids.
• 4.4. Residual triglycerides contained 80% of saturated and monoenoic fatty acids.
• 5.5. Linoleic acid increased from 2% in feed TG to 10% in TG of the rectum.

     

Properties of enzymes hydrating epoxides in human epidermis and liver

• 1.1. Cytosolic and microsomal epoxide hydrolyzing enzymes of human skin and liver were compared and found to be different.
• 2.2. Epidermal and hepatic cytosolic epoxide hydrolases were different in terms of substrate selectivity, pI, inhibitor sensitivity and affinity Chromatographic properties.
• 3.3. Microsomal epoxide hydrolases had the same pIs but different substrate selectivities.
• 4.4. Cytosolic epoxide hydrolase from adults had higher specific activity than that from neonates or cultured epidermis, but lower activity than adult hepatic enzymes.
• 5.5. The sizes of cytosolic epoxide hydrolase from epidermis and liver were similar and lower than that from cultured fibroblasts.
• 6.6. Cytosolic epoxide hydrolase from all sources shared similar antigenic determinants.

     

Hematology and blood chemistry of the marsh harrier (Circus aeruginosus)

• 1.1. This study deals with the hematological and blood chemistry of 13 adult marsh harriers (Circus aeruginosus).
• 2.2. No significant differences were observed between male and female groups in any of the parameters.
• 3.3. The value of white blood cells was 14,677/mm³ heterophils and lymphocytes, these being the most abundant cellular type (81.42 and 12%, respectively).
• 4.4. Urea and uric acid are present in approximately similar proportions, though birds are said to be uricotelic.
• 5.5. The cholesterol concentration values determined in our study, are higher than those reported in most other birds.

     

Drosophila mettleri and senita cactus alkaloids: Fitness measurements and their ecological significance

• 1.1. Drosophila mettleri have been found feeding but not breeding on decaying stems of senita cactus, the normal host for Drosophila pachea.
• 2.2. Alkaloids were extracted from senita stems and used in tests of egg-to-adult viability, developmental rate, and adult longevity.
• 3.3. The results show that developmental rate is not appreciably affected by senita alkaloids.
• 4.4. In general, D. mettleri was less affected by the alkaloids with respect to egg-to-adult viability and adult longevity than D. pachea at concentrations which are fatal to other desert Drosophila.
• 5.5. Tolerance to alkaloids gives D. mettleri an ecological advantage.

     

Partial characterization of the hemoglobin from Hubbs' beaked whale (Mesoplodon carlhubbsi)

• 1.1. Blood samples were obtained from an adult female Hubbs' beaked whale and her fetus.
• 2.2. Two major hemoglobins were demonstrated by cellulose acetate electrophoresis and were purified by ion exchange chromatography from each specimen.
• 3.3. The relative amounts of these components were different in the adult and fetus.
• 4.4. Both of these hemoglobins have a higher affinity for oxygen than normal human hemoglobin.
• 5.5. Maternal and fetal hemoglobins were separated and the N-terminal amino acid of each of these hemoglobins was found to be valine.
• 6.6. Tryptic peptide separation and amino acid analyses of the purified polypeptide chains indicated that the hemoglobins of the fetal sample were identical to those of the maternal.

     

Proteomic Analysis of Vocal Fold Fibroblasts Exposed to Cigarette Smoke Extract: Exploring the Pathophysiology of Reinke's Edema,

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Highlights

• •First global study on laryngeal cells cultured with cigarette smoke enriched medium.
• •Vocal fold fibroblasts increased production of ECM component hyaluronan.
• •Expression of several fibrillar collagens was reduced.

     

A comparative study on the biological properties of venoms from juvenile and adult common tiger snake (Notechis scutatus) venoms

• 1.1. The biological properties of venoms from juvenile and adult common tiger snakes (Notechis scutatus) were compared.
• 2.2. The lethality, procoagulant activity and enzymatic activities of the juvenile venom were not substantially different from those of the adult venom.
• 3.3. Electrophoretic studies, however, indicated some minor differences in the protein composition of the juvenile and adult venoms.

     

Tritiated water measurements of water flux in nestling and adult zebra finches (Poephila guttata)

• 1.1. Total body water volume (TBWV) and daily water turnover rates (WTR) were measured in nestling and adult zebra finches using tritiated water (TOH).
• 2.2. TBWV and daily WTR increased with age up to 13 days post hatching.
• 3.3. TBWV and WTR approached adult levels after 13 days of age. WTR varied among ages and nest mates. All nestlings turned over at least 50% of their body water pool per day.
• 4.4. The WTR data for adult birds are consistent with natural history data suggesting zebra finches are dependent on water for breeding and survival.

     

Identification of Novel Natural Substrates of Fibroblast Activation Protein-alpha by Differential Degradomics and Proteomics

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Highlights

• •Natural substrates of FAP were identified using degradomic and proteomic techniques and FAP gene knockout mouse derived embryonic fibroblasts stably transduced with enzymatically active or inactive FAP.
• •Terminal amine isotopic labelling of substrates (TAILS) based degradomics identified cleavage sites in collagens, and many other extracellular matrix (ECM) and associated proteins.
• •Cleavages of lysyl oxidase-like-1, CXCL-5, CSF-1 and C1qT6 by FAP were confirmed in vitro.
• •Differential metabolic labelling coupled with quantitative proteomic analysis implicated FAP in regulating proteins that are associated with ECM, ECM-cell interactions, coagulation, metabolism and wound healing.

     

The high intracellular ph buffering of hagfish (Eptatretus cirrhatus) dental plate retractor muscle cannot be attributed to the known histidine-related compounds present in other vertebrates

• 1.1. Intracellular pH buffering capacity of hagfish (Eplatretus cirrhatus) dental plate retractor muscles is among the highest reported for any vertebrate muscle.
• 2.2. Over 80% of the pH buffering capacity of hagfish retractor and myotome muscle is due to components other than proteins and phosphate.
• 3.3. The muscles have less than 0.5 μmol/g wet weight of l-histidine, and lack l-l-methyl histidine, l-3-methyl histidine and the histidine-containing dipeptides anserine, carnosine and ophidine.
• 4.4. Instead, they contain an unidentified low molecular weight acid-soluble compound to which the high pH buffering capacity can be attributed.