O2-binding by the blood of the landhopper,Arcitalitrus dorrieni (Hunt) (Crustacea: Amphipoda)

• 1.1. The O₂-binding characteristics of the blood of the euterrestrial amphipod (landhopper) Arcitalitrus dorrieni have been studied.
• 2.2. The blood exhibited a low O₂ affinity, with a p₅₀ (at pH = 7.8) of 21.4 torr (10°C). Affinity decreased with an increase in temperature at constant pH (ΔH = − 79.4kJ/mol) but the Bohr factor (ΔlogP₅₀/Δ pH = −0.67) was unaffected.
• 3.3. The O₂-carrying capacity of the blood was moderate (1.51 ml/100 ml)
• 4.4. The results support the hypothesis that the blood of terrestrial amphipods is characterized by having a low affinity pigment.

     

Lysosomal acid deoxyribonuclease from vervet monkey livers—II: Enzymic properties

• 1.1. Primate liver lysosomal acid DNase is an endonucleolytic enzyme.
• 2.2. The enzyme has both 3'- and 5'-nucleotidohydrolase activities.
• 3.3. The oligonucleotides produced by DNase are polymers mainly about 30 mononucleotides long.
• 4.4. The Arrhenius plot shows a discontinuity with a transition temperature at 47°C, with an activation energy of 107 kJ/mol below and 67 kJ/mol above this temperature.
• 5.5. The activation enthalpy is 104kJ/mol and the entropy −0.498 kJ/mol/K.
• 6.6. The enzyme is subject to substrate inhibition and the K_m value is 159 × 10⁻³mM DNA-P.

     

The effect of temperature on the acid-base status of the blood of the hatching quail (Coturnix c. japonica)

• 1.1. The ambient temperature of embryos of pipped eggs was reduced from 38 to 28°C for a period of 45 min.
• 2.2. The blood P_CO2 was lower and the blood more alkaline at 28°C than at 38°C.
• 3.3. At 28°C plasma [HCO₃⁻] ] was lower than predicted from the blood buffer line determined in vitro.
• 4.4. The plasma concentrations of strong ions and lactate were the same at both temperatures.
• 5.5. After the ambient temperature had been returned to 38°C for a period of 45 min, blood pH was more acidic than before cooling, but there was no difference in blood P_CO2.
• 6.6. The plasma [HCO₃⁻] was the same as that at 28°C and plasma [K⁺] was higher than before cooling.
• 7.7. The results arc discussed in relation to the factors affecting blood pH in embryos at this stage of development.

     

The dynamics of the permeation of an analogue of insect juvenile hormone into nematodes

• 1.1. ¹⁴C-dichlorofarnesoate permeated rapidly into Haemonchus contortus (infective juveniles) and Panagrellus redivivus (mixed cultures) and was strongly bound by hydrophobic association (K_s > 10⁻⁴M).
• 2.2. Uptake rose linearly with increases in temperature (5–38°C) and external concentration (C₀; 0.07–2.15 × 10⁻⁴ M). Within 1 hr the internal concentration, C₁ was >C C₀.
• 3.3. The pH of the medium (6–8) did not affect uptake.
• 4.4. Efflux of dichlorofarnesoate was low: the half-time of release was > 18 hr.
• 5.5. The uptake curve approximated to the expression C₁/C₀ = a(1 − e^−bt) with a and b as constants and t in hr.
• 6.6. These results clarify previous work on the inhibitory action of juvenile hormone on the development of nematodes.

     

Depressed baroreceptor-cardiac reflex sensitivity during simulated diving in ducks

• 1.1. The baroreceptor-cardiac reflex was examined in unanesthetized ducks at rest and during diving. In ducks breathing air an inverse relationship between mean arterial blood pressure and heart rate was observed over the pressure range from 80cm H₂O to 290cm H₂O.
• 2.2. Increases in pressure were obtained by bolus intravenous injection of phenylephrine (PE) while the hypotension was achieved by intravenous acetylcholine (ACh).
• 3.3. The inverse relation of blood pressure and heart rate was also observed in ducks without pharmacologic intervention.
• 4.4. The baroreceptor-cardiac reflex sensitivity was -3.13 beats/min/cm H₂O in non-diving ducks and fell to -0.96 beats/min/cm H₂O with PE and ACh derived data.
• 5.5. During diving the baroreceptor-cardiac reflex sensitivity was further reduced to —0.48beats/ min/cm H₂O.
• 6.6. This finding indicates that even during the pronounced bradycardia of diving baroreceptor stimulation continues to influence heart rate.

     

Effects of water stress on hemolymph volume,osmotic potential and chemical composition in Megetra cancellata

• 1.1. Rates of water loss in Megetra cancellata were very high compared to those reported for other xeric arthropods.
• 2.2. Hemolymph weight in hydrated animals was 43.0% of the total body weight while it was 24.7% in desiccated animals that had lost 16.1% of their body weight as water.
• 3.3. Hemolymph osmotic potential increased from 417 to 447 mOsm/kg in desiccated beetles, but osmotic regulation was evident.
• 4.4. Total hemolymph protein mass and concentration decreased in desiccated beetles while amino acid concentrations remained constant (at about 70 mM).
• 5.5. Na⁺ and −PO₄ concentrations increased in desiccated beetles.
• 6.6. Cl⁻ and K⁺ concentrations in desiccated beetles were equal to those in undesiccated beetles.

     

A comparative study of the respiratory properties and physiological function of haemocyanin in two burrowing and two non-burrowing crustaceans

• 1.1. Oxygen dissociation curves were constructed for the haemolymph of two non-burrowing, Galathea strigosa and Eupagurus bernhardus, and two burrowing crustaceans, C. cassivelaunus and Nephrops norvegicus. The p₅₀ at in vivo pH values and 10°C was 12.6 Torr in G. strigosa, 23 Torr in E. bernhardus, 3.1 Torr in C. cassivelaunus and 11.5 Torr in N. norvegicus.
• 2.2. The Bohr values (Δlogp₅₀/ΔpH) were high in all species ranging between −0.96 and −1.48. Cooperativity expressed as P₅₀ averaged 3.3, 3.8 and 3.8 in G. strigosa, E. bernhardus and N. norvegicus. respectively. A lower value of 2.2 was observed in C. cassivelaunus.
• 3.3. The oxygen affinity of the haemocyanin was relatively temperature independent, the values for ΔH at pH7.9 ranging between −5.1 and −18.1 kJmol⁻¹.
• 4.4. Haemolymph respiratory gas analysis showed values similar to those previously reported in crustaceans: p_aO₂ ranging between 44 and 107 Torr and p_vO₂ values between 18 and 24 Torr.
• 5.5. Pre-/post-branchial pH differences were small in G. strigosa, E. bernhardus and N. Norvegicus, but averaged 0.09 of a pH unit in C. cassivelaunus. p_aCO₂ and P_vCO₂ values ranged between 1.4 and 2.3 Torr.
• 6.6. In buried C. cassivelaunus both pre- and post-branchial oxygen tensions decreased, as did oxygen tension overall during respiratory pauses.
• 7.7. Cardiac output values were low, ranging between 59 and 71 ml kg⁻¹ min⁻¹ for all four species and calculated stroke volumes were realistic in terms of animal size.
• 8.8. In the non-burrowing species physically dissolved oxygen accounted for 5–21% of the oxygen transported to the tissues. In the burrowing species values of 40–77% were found.

     

Characteristics of Crassostrea virginica crystalline style chitin digestion

• 1.1. Fundamental chitin digestion characteristics of Crassostrea virginica crystalline style were investigated.
• 2.2. Optimum temperature and pH were 34°C and 4.8. respectively.
• 3.3. The colloidal regenerated chitin (0.56mol/0.5 ml: GlcNAc equivalents) was saturating under all enzyme levels encountered.
• 4.4. There was no evidence of end product inhibition, even after 100 hr incubation.
• 5.5. Calculated K_m for the chitinase complex was 1.19mM when determined using a 30 min assay, but was only 0.70 mM when determined using a 4.6 hr assay.
• 6.6. Both K_m values are lower than reported for similar assays in other molluscs and for most bacteria.
• 7.7. Effect of substrate preparation on the kinetics are discussed.
• 8.8. Eight peaks of chitinase activity were resolved by DEAE-Fractogel ion exchange chromatography.

     

Electrical transport characteristics of Aplysia californica intestinal epithelium

• 1.1. Replacing chloride (Cl⁻) with sulfate (SO₄²⁻) in the bathing medium drastically reduced the mucosal membrane potential difference (ψ_m).
• 2.2. The voltage divider ratio was significantly greater than one.
• 3.3. Mucosal ^d-glucose decreased the input resistance of the intestinal epithelium.
• 4.4. Addition of furosemide to the mucosal bathing medium inhibited transepithelial potential difference and short-circuit current.
• 5.5. Addition of SITS to the mucosal bathing medium partially inhibited transepithelial potential difference and short-circuit current.
• 6.6. Diffusion potentials in the intestinal epithelium were symmetrical.

     

The effect of temperature on oxygen equilibrium curves of trout blood (Salmo gairdnerii)

• 1.1. Oxygen equilibrium curves were measured on trout red blood cell suspensions at pH 7.8 and 8.4 at 15, 20 and 25 C. Normal red cells and red cells that had been depleted of their ATP content were used.
• 2.2. The equilibrium data were fitted to the Adair's model and the enthalpy (ΔH) and entropy (ΔS) changes for the first and fourth steps of oxygenation and for overall oxygenation were calculated from the temperature dependencies of the Adair constants.
• 3.3. For normal red blood cells, the apparent heat for the first oxygenation step, δh₁, is close to zero.
• 4.4. Temperature insensitivity of this step at physiological pH, combined with a large pH dependence, probably denotes a property of Hb4, the Root effect Hb of trout blood.
• 5.5. At pH 7.8, ΔH₄ is about —4kcal/mol, a small value which may be attributed to the large release of Bohr protons that occurs at the last oxygenation step and corresponds to an endothermic process which opposes to the exothermic oxygenation of the haem.
• 6.6. The ΔH₄ value appears to have a large influence on the enthalpy for overall oxygenation.
• 7.7. Results for ATP-free red cells are consistent with a mere increase in the intracellular pH and suggest that ATP has no specific effect at and above pH_i ~ 7.7.
• 8.8. Effects of temperature and pH on trout red blood cell isotherms emphasize the primary importance of the major component of trout blood, namely Hb4, in trout blood functional properties.

     

Preparation of two neurotoxic esterases from the chick central nervous system

• 1.1. A standard procedure for lipid-extraction of lyophilized hen brain material is decribed.
• 2.2. Nine carboxylesterase isoenzymes (EC 3.1.1.1) are identified in lipid-extracted lyophilized material (LELM) using kinetic analysis of organophosphate inhibition. Total phenyl valerate (PV) hydrolysing carboxylesterase activity in LELM is 43.3U.g⁻¹
• 3.3. Two carboxylesterase isoenzymes of LELM are classified as neurotoxic esterases (NTE_A and NTEg_B).
• 4.4. Using n-octylglucoside 51% of the water-insoluble neurotoxic esterase activity from LELM are solubilized.
• 5.5. Six carboxylesterase isoenzymes including NTE_A (6.5 U-l⁻¹) and NTE_B (4.2 U-l⁻¹) are present in the solubilized preparation.
• 6.6. Throughout purification and separation steps carboxylesterase isoenzymes are identified by their rate constants for the reaction with organophosphorus inhibitors.

     

The pattern and influential factors of aquatic pharyngeal movements of Trionyx sinensis

• 1.1. The pharyngeal movements of Trionyx sinensis during submersion where recorded with physiological instruments.
• 2.2. Anoxia or hypercapnia caused a marked increase in breathing rate of tested turtles during voluntary diving, and in anoxia there was a significant increase in the frequency of aquatic pharyngeal movements while hypercapnia had a slight or no effect on the frequency of these movements.
• 3.3. During voluntary diving when turtles could easily extend their heads out of water to breathe air, the frequency of rhythmic pharyngeal movements was lower; but during forced submersion, the frequency was higher and the movements were continuous.
• 4.4. The frequency increased more rapidly and greatly when turtles were in forced submersion than when they dived freely and could easily surface to breathe in N₂.
• 5.5. The frequency of pharyngeal movements of T. sinensis during diving in an aquarium with water depth of 30 or 45 cm was markedly higher than that at a water depth of 15 cm. Disturbing stimuli also influenced the aquatic rhythmic pharyngeal movements of T. sinensis.

     

Gas tensions within the egg of the quail (Coturnix c. Japonica) during the closing stages of embryonic development

• 1.1. The respiratory status of the embryonic quail during the two days prior to hatching was assessed by measuring the gas tensions within the air space of the egg and of blood collected from the chorioallantois.
• 2.2. When the lungs became inflated there was a significant decrease in the pO₂ of the gas in the air space.
• 3.3. After pipping, there was a rise in the pO₂ and fall in the pCO₂ within the air space, together with corresponding changes in the blood.
• 4.4. The outer shell membrane remained intact until the onset of hatching.
• 5.5. These results were compared with those obtained by other workers using the domestic fowl.

     

Metabolism of exogenous uridine 5'-triphosphate,adenosine 5'-triphosphate and pyrophosphate by alkylsulfatase-producing bacteria

• 1.1. The uptake and metabolism of β-P³²-labelled UTP, ATP and uniformly labelled PPi was studied using bacteria washed twice with and immediately resuspended in a medium containing 0.031 M PO³⁻₄.
• 2.2. Under these conditions, little P³²-labelled PO³⁻₄ was detected extracellularly when the cells were exposed for 2–5 min to 5 × 10⁻⁵ M concentrations of the above effectors.
• 3.3. Cells of both Pseudomonas aeruginosa and Pseudomonas C₁₂B incorporated the P³²-label from all effectors under the above conditions.

     

Proteolytic cleavage of band 3 protein in relation to anion transport in fish (Oncorhynchus mykiss) red blood cells

• 1.1. The effects of trypsin and chymotrypsin on HCO₃⁻/Cl⁻ exchange through red blood cell membranes of humans and trout were studied.
• 2.2. To measure the anion exchange we used a right-angle light-scattering technique by applying the Jacobs-Stewart cycle in ammonium solution and the osmotiration method at constant cell volume.
• 3.3. The Cl⁻ flux in human red blood cells remained unaltered after treatment with external trypsin and chymotrypsin while in trout red blood cells the flux decreased.
• 4.4. This partial inhibition of anion transport in fish, ranging from 30 to 40%,suggest that one or several of the cleavage sites in band 3 protein, essential for anion transport function, are exposed in fish red blood cells.
• 5.5. In human red blood cells the fragments of band 3 which are affected by proteolytic digestion, retain their tertiary structure because there is no influence on anion transport.

     

Evidence for hyporegulation in the calanoid copepod,Acartia tonsa

• 1.1. The euryaline calanoid copepod, Acartia tonsa, maintains haemolymph Na below that of the external medium in salinities above 34^o_oo (475 mM Na).
• 2.2. The measured transepithelial electrical potential. −9.97 ± 1.0 mV, indicates that Na is regulated out of electrochemical equilibrium.
• 3.3. Water osmotically lost in hyporegulation is replaced by Na-dependent absorption by the gut.
• 4.4. High osmotic water permeability is evidenced by the fact that with an increase in external salinity from 475 mM Na to 580 mM Na the copepod's drinking rate nearly doubles.
• 5.5. Sodium efflux measurements indicate that ionic permeability is much lower than other hyporegulating crustaceans.
• 6.6. The energetic advantage of hyporegulation in this species is considered.

     

The oxygen consumption rates of three gastrotrichs

• 1.1. The oxygen consumption rates for three sympatric species of marine gastrotrichs (anatomically similar, except that one contains hemoglobin) were measured with a Cartesian diver microrespirometer.
• 2.2. The rates for the two species without hemoglobin, Turbanella ocellata and Dolichodasys carolinensis, were 307.2 μl O₂ g⁻¹ hr⁻¹ and 108.0 μl O₂ g⁻¹ hr⁻¹, respectively, while the rate for the hemoglobin-containing species, Neodasys, was 208.9 μl O₂ g⁻¹ hr⁻¹.
• 3.3. The possession of hemoglobin by Neodasys (14% by volume) cannot be explained by an unusually high demand for oxygen.
• 4.4. Instead, the hemoglobin may be useful as an oxygen store providing continued aerobic metabolism in anoxic conditions, thus allowing Neodasys to exploit a different niche.

     

Human placental atp-diphosphohydrolase: Biochemical characterization,regulation and function

• 1.1. Kinetic and physico-chemical studies on human placental microsomal fraction confirmed that the ATPase and ADPase activities detected in this fraction correspond to the enzyme ATP-diphosphohydrolase or apyrase (EC 3.6.1.5). These include substrate specificity, and coincident M_r and pI values of both ATPase-ADPase activities.
• 2.2. This enzyme hydrolyses both the free unprotonated and cation-nucleotide complex, the catalytic efficiency for the latter being considerably higher.
• 3.3. Microsomal apyrase is insensitive to ouabain and Ap5A. The highly purified enzyme was only inhibited by o-vanadate, DBS and slightly by DCCD.
• 4.4. Apyrase seems to be a glycoprotein from its interaction with Concanavalin-A.
• 5.5. Preliminary studies on the essential amino acid residues suggest the participation of Arg, Lys and His residues, and discard the requirement of −SH, COO⁻, −OH, and probably also Tyr and Trp.
• 6.6. Two kinetic modulatory proteins of apyrase were detected in placental tissue. An activating protein was found in the soluble fraction and an inhibitory protein was loosely bound to the membranes.
• 7.7. The proposed in vivo function for apyrase is related to the inhibition of platelet aggregation due to its ADPase activity, which is supported by the direct effect on washed platelets and by its plasma membrane localization.

     

l-lysinamidase from Cryptococcus laurenti 112. Purification and properties

• 1.1. The purified enzyme hydrolyzes the linear l-lysinamide and the cycle amide of l-lysine—l-α-amino-ϵ-caprolactam.
• 2.2. The apparent relative molecular mass is 180,000. The enzyme consists of four subunits and the molecular mass of a single subunit was found to be 47,000.
• 3.3. The coefficient of molecular sedimentation equals 8.3 S, the isoelectric point was determined to be pH 4.3
• 4.4. The enzyme is not a glycoprotein. p-Mercuribenzoate binds 10 SH-groups of the native enzyme molecule and 20 SH-groups in the presence of 0.7% SDS.
• 5.5. pH- optimum for the hydrolysis of l-lysine amides was observed to be 7.5–7.7. The enzyme is strictly dependent on Mn²⁺ and Mg²⁺.
• 6.6. The kinetic parameters for the hydrolysis of l-lysinamide where K_m = 3.8 mM and k_cat = 3000 sec⁻¹ For the hydrolysis of cyclic L-lysinamide K_m = 4.8 mM and k_cat = 2600 sec⁻.

     

Oxygenation characteristics of the polymorphic hemoglobins of coho salmon (Oncorhynchus kisutch) at different developmental stages

• 1.1. The oxygen equilibria of the multiple hemoglobin of the fry and adult life stages of coho salmon (Oncorhynchus kisutch) were investigated. Components A 6–8 comprise 50–55% and >95% of the total hemoglobins of coho adults and fry respectively.
• 2.2. Fry hemoglobins exhibited a high oxygen affinity (P₅₀ = 3.9 mm Hg at 9.8°C, pH 8.5), a very large Bohr shift (ø = Δlog ₅₀/ΔpH = −1.729 at pH 7.1–7.5) which was non-linear in the pH range 6.8–8.5 and a large heat of oxygenation (ΔH = −20.8 kcal/mol).
• 3.3. Adult hemoglobins, however, exhibited a moderate oxygen affinity (P₅₀ = 14.0 mm Hg at 9.8°C, pH 8.2), a very small linear Bohr shift (ø = −0.172 at pH 7.0 −8.2) and a low heat of oxygenation (ΔH = −5.6- −7.5kcal/mol). Adenosine triphosphate had only minor influence on the oxygenation characteristics of adult heolyzates.
• 4.4. The results are discussed in terms of functional adaptation to environmental stresses and metabolic requirements.