Penicillin degradation catalysed by Zn(II) ions in methanol

The rates of degradation, catalysed by Zn²⁺, of four classical penicillins—amoxicillin, ampicillin and penicillins G and V—were followed at 20 °C in methanol by spectrophotometric assays. Kinetic schemes of the reactions of degradation catalysed by Zn²⁺ ions were analogous to those given previously for the reaction catalysed by Cd²⁺ ions. The methanolysis of penicillin V occurs with the formation of a single intermediate substrate–metal complex (SM), whereas the degradations of amoxicillin, ampicillin and penicillin G occur with the initial formation of two complexes with different stoichiometry, SM and S₂M, both in equilibrium. In all cases, the degradation reaction is of the first order with respect to SM, with velocity constants at 20 °C of 0.0093, 0.0288, 0.0304 and 0.0349 min⁻¹, for amoxicillin, ampicillin, penicillin V and penicillin G, respectively. The compound S₂M degraded at a much lower rate than SM and constitutes a zero-order process. The catalytic effect of the ion Zn²⁺ in the degradation of the penicillins was much weaker than that of the ion Cd²⁺, owing to the lesser ionic radius of the former and the fact that in the case of the reaction catalysed by Zn²⁺, the compound S₂M occurred in a much greater amount than the SM. At the end of the degradation reaction, the corresponding penamaldic derivative of the antibiotic was produced, established by the coordination of the Zn²⁺ ion, forming a single complex 2:1 (derivative penamaldic–metal) in the case of amoxicillin and ampicillin; and two complexes, 1:1 and 2:1, for the other antibiotics. Finally, the molar absorption coefficients of the products of reaction at the wavelength of maximum absorption at 20 °C were calculated.     

Influence of phosphorus concentration on the growth kinetics and stoichiometry of the microalga Scenedesmus obliquus

The growth of the freshwater microalga Scenedesmus obliquus was studied at 30°C in a mineral culture medium with phosphorus concentrations of between 0 and 372 μ . The values for the specific growth rates, between and , fitted a semistructured substrate-limitation model with μ_m1 = 0·0466 h⁻¹, μ_m2 = 0·0256 h⁻¹ and . The specific uptake rate of phosphorus reached a maximum value of q_Sm1 = 658·01 × 10⁻⁴ μmol P mg⁻¹ biomass h⁻¹.     

The use of chemically modified and unmodified cassava waste for the removal of Cd, Cu and Zn ions from aqueous solution

The use of different chemically modified cassava waste biomass for the enhancement of the adsorption of three metal ions Cd, Cu and Zn from aqueous solution is reported in this paper. Treating with different concentrations of thioglycollic acid modified the cassava waste biomass.

The sorption rates of the three metals were 0.2303 min⁻¹ (Cd²⁺), 0.0051 min⁻¹ (Cu²⁺), 0.0040 min⁻¹ (Zn²⁺) and 0.109 min⁻¹ (Cd²⁺), 0.0069 min⁻¹ (Cu²⁺), 0.0367 min⁻¹ (Zn²⁺) for 0.5 and 1.00 M chemically modified levels, respectively. The adsorption rates were quite rapid and within 20–30 min of mixing, about 60–80% of these ions were removed from the solutions by the biomass and that chemically modifying the binding groups in the biomass enhanced its adsorption capacity towards the three metals. The results further showed that increased concentration of modifying reagent led to increased incorporation, or availability of more binding groups, in the biomass matrix, resulting in improved adsorptivity of the cassava waste biomass. The binding capacity study showed that the cassava waste, which is a serious environmental nuisance, due to foul odour released during decomposition, has the ability to adsorb trace metals from solutions.     

Pyrolysis of safflower (Charthamus tinctorius L.) seed press cake: part 1. The effects of pyrolysis parameters on the product yields

Safflower (Charthamus tinctorius L.) seed press cake was pyrolysed in a fixed-bed reactor. The effects of pyrolysis temperature, heating rate and sweep gas flow rates on the yields of the products were investigated. Pyrolysis runs were performed using pyrolysis temperatures between 400 and 600 °C with heating rates of 10, 30 and 50 °C min⁻¹. The obtained bio-char, gas and bio-oil yields ranged between 25 and 34 wt%, 19 and 25 wt%, and 28 and 36 wt%, respectively, at different pyrolysis conditions. The highest liquid yield was obtained at 500 °C pyrolysis temperature with a heating rate of 50 °C min⁻¹ under the sweep gas of N₂ with a flow rate of 100 cm³ min⁻¹. Employing the higher heating rate of 50 °C min⁻¹ results in maximum bio-oil yield, probably due to the decrease in mass transfer limitations. According to the results obtained under the conditions of this study, the effects of pyrolysis temperature and sweep gas flow rate are more significant than the effect of heating rate on the yields.     

Intramuscular temperatures during exercise in the heat following pre-cooling and pre-heating

Pre-cooling improves heat tolerance and time to exhaustion in the heat. We tested the possibility that reduced tissue temperatures may explain this phenomenon, using three whole-body treatments: pre-cooling, thermoneutral (control) and pre-heating. Pre-cooling reduced muscle temperature (T_m) by 6.3 °C while pre-heating increased T_m 3.4 °C, relative to control. Despite this offset, T_m climbed towards a common asymptote, with pre-cooling offering no thermal protection beyond 40 min. Following pre-cooling, exercising oesophageal temperature (T_es) initially increased at 0.09 °C min⁻¹, being significantly faster than control (0.05 °C min⁻¹) and pre-heated conditions (0.03 °C min⁻¹). Pre-cooling lowered the sweat threshold and also resulted in a reduced cardiac frequency across the exercise-heat exposure. Our observations do not support the hypothesis that pre-cooling reduces T_m at the end of an exercise-heat exposure, thereby delaying the development of fatigue.     

Different thermal sensitivity to exercise and cold in men and women

Rectal temperature gain for sweating and shivering was investigated in men (M) and women during exercise and cold exposure. Women, taking and not taking oral contraceptives, were tested during follicular (F) and luteal (L) phases of the menstrual cycle. The gains for sweating (M, 4.85; F, 5.00; L, 12.25 kPa °C⁻¹) and shivering (M, 6.50; F, 2.75; L, 2.00 ml kg min⁻¹ °C⁻¹) differed between sexes, as well as between F and L, and were affected by the oral contraceptives (5.45 and 8.00 kPa °C⁻¹, P<0.01, and 6.00 and 4.25 ml kg min⁻¹ °C⁻¹, P<0.05, respectively).     

Thermodynamics of the disproportionation of adenosine 5'-diphosphate to adenosine 5'-triphosphate and adenosine 5'-monophosphate, II. Experimental data

High-pressure liquid-chromatography and microcalorimetry have been used to determine equilibrium constants and enthalpies of reaction for the disproportionation reaction of adenosine 5′-diphosphate (ADP) to adenosine 5′-triphosphate (ATP) andadenosine 5′-monophosphate (AMP). Adenylate kinase was used to catalyze this reaction. The measurements were carried out over the temperature range 286 to 311 K, at ionic strengths varying from 0.06 to 0.33 mol kg⁻¹, over the pH range 6.04 to 8.87, and over the pMg range 2.22 to 7.16, where pMg = -log a(Mg²⁺). The equilibrium model developed by Goldberg and Tewari (see the previous paper in this issue) was used for the analysis of the measurements. Thus, for the reference reaction: 2 ADp³⁻ (ao) AMp²⁻ (ao)+ ATp⁻ (ao), K° = 0.225 ± 0.010, ΔG° = 3.70 +- 0.11 kJ mol ⁻¹, ΔH° = −1.5 ± 1. 5 kJ mol ⁻¹, °S ° = −17 ± 5 J mol⁻¹ K⁻¹, and ACP_p°≈ = −46 J mo1l⁻¹ K⁻¹ at 298.15 K and 0.1 MPa. These results and the thermodynamic parameters for the auxiliary equilibria in solution have been used to model the thermodynamics of the disproportionation reaction over a wide range of temperature, pH, ionic strength, and magnesium ion morality. Under approximately physiological conditions (311.15 K, pH 6.94, [Mg²⁺] = 1.35 × 10⁻³ mol kg⁻¹, and I = 0.23 mol kg⁻¹) the apparent equilibrium constant (K_A′ = m(ΣAMP)m(ΣATP)/[ m(ΣADP)]²) for the overall disproportionation reaction is equal to 0.93 ± 0.02. Thermodynamic data on the disproportionation reaction and literature values for this apparent equilibrium constant in human red blood cells are used to calculate a morality of 1.94 × 10⁻⁴ mol kg⁻¹ for free magnesium ion in human red blood cells. The results are also discussed in relation to thermochemical cycles and compared with data on the hydrolysis of the guanosine phosphates.     

Graft copolymerization of acrylic acid onto guar gum initiated by vanadium (V)–mercaptosuccinic acid redox pair

Guar gum has been modified by graft copolymerization with acrylic acid in aqueous medium using vanadium (V)–mercaptosuccinic acid redox system. The optimum reaction conditions affording maximum grafting ratio, efficiency, add on and conversion have been determined. The grafting parameters have been found to increase with increase in vanadium (V) concentration upto 1.0 × 10⁻² mol dm⁻³, but these parameters decrease on further increasing the vanadium (V) concentration. On increasing the mercaptosuccinic acid concentration from 1.0 × 10⁻² to 4.0 × 10⁻² mol dm⁻³ grafting ratio, efficiency and add on increase up to 2.0 × 10⁻² mol dm⁻³ but decrease with further increase in mercaptosuccinic acid concentration. On varying the acrylic acid concentration from 5.0 × 10⁻² to 30.0 × 10⁻² mol dm⁻³, maximum grafting ratio, efficiency and add on have been obtained at 20.0 × 10⁻² mol dm⁻³. The grafting ratio, add on and conversion increase, on increasing the H⁺ ion concentration from 1.5 × 10⁻¹ to 6.0 × 10⁻¹ mol dm⁻³. On increasing the guar gum concentration the grafting parameters increase. The grafting ratio, add on and conversion have been found to increase with time period while efficiency started decreasing after 120 min. It has been observed that %G increases on increasing the temperature up to 35 °C. The graft copolymer has been characterized by IR spectroscopy and thermogravimetric analysis.     

In-vitro heat exchange at the rete of the Boer goat under specified laboratory conditions

1. Each half rete from five Boer goats was perfused with water at 38 °C and flow rate of 2 ml min⁻¹ while simultaneously perfusing the cavernous sinus with water at different temperatures and flow combinations and recording temperatures across the rete.

2. The minimum temperature difference across the rete was recorded at a cavernous sinus perfusion temperature of 37.8 °C and flow rate of 2 ml min⁻¹.

3. Slopes of heat exchange increased threefold when the flow was increased four times.

4. These results support the idea that the rete is an obligate heat exchanger.

Hydrodynamic properties of the fractions of mannan formed by Rhodotorula rubra yeast

Aqueous solutions of fractions of an extracellular linear mannan formed by Rhodotorula rubra yeast have been investigated by hydrodynamic methods (high-speed sedimentation, translation isothermic diffusion and viscometry). The molecular weight was determined according to Svedberg ( ) and the polydispersity parameters of the initial sample were also determined (M_w/M_n = 1·20 and M_z/M_w = 1·21). Relationships between the molecular weight (M) and s_o, D_o and [η] in the range were: [η] = 2·33 × 10⁻² M^0.75, D_o = 1·65 × 10⁻⁴ M^0·58, s_o = 2·24 × 10⁻¹⁵ M^0·43. The equilibrium rigidity and hydrodynamic diameter of chains representing mannan molecules were evaluated.     

Cholesterol sulphate sulphohydrolase from human placenta microsomes--purification and properties of the dephosphorylated form of enzyme

The procedure for purification of cholesterol sulphate sulphohydrolase (ChS-ase) from human placenta microsomes was elaborated. The highy purified enzyme preparation (specific activity 2000 nmol×min⁻¹×mg protein⁻¹) exhibited optimal activity at pH 9.0. The K_m value was established to be 1.5±0.85×10⁻⁵ M. The high molecular weight form (200 kDa) and the low molecular weight form (20 kDa) of the enzyme were separated. The interconversion of the high molecular weight variant into the low one occurs under the influence of dephosphorylation. Both forms exhibited typical Michaelis–Menten saturation kinetics. The effect of different compounds on the enzyme activity was tested.     

Large-scale preparation of a nitrile-hydrolysing biocatalyst: Rhodococcus R 312 (CBS 717.73)

Lyophilized cells of Rhodococcus R 312 (CBS 717.73) can be employed as an easy-to-use biocatalyst for the biocatalytic hydrolysis of nitriles to furnish the corresponding carboxamides or acids on a preparative scale. The following practical aspects are advantageous: (i) Fermentation yields a high biomass (10 g dry cell weight l⁻¹), (ii) enzyme induction is not required, (iii) a maximum of activity is obtained at the late exponential growth phase (25.7 μmol min⁻¹ mg⁻¹), which can be monitored by a simple photometic assay and (iv) the cells can be stored at +4°C for several months without significant loss of activity.     

Photosynthetic response of Myriophyllum salsugineum A.E. orchard to photon irradiance, temperature and external free CO2

The photosynthetic capacity of Myriophyllum salsugineum A.E. Orchard was measured, using plants collected from Lake Wendouree, Ballarat, Victoria and grown subsequently in a glasshouse pond at Griffith, New South Wales. At pH 7.00, under conditions of constant total alkalinity of 1.0 meq dm⁻³ and saturating photon irradiance, the temperature optimum was found to be 30–35°C with rates of 140 μmol mg⁻¹ chlorophyll a h⁻¹ for oxygen production and 149 μmol mg⁻¹ chlorophyll a h⁻¹ for consumption of CO₂. These rates are generally higher than those measured by other workers for the noxious Eurasian water milfoil, Myriophyllum spicatum L., of which Myriophyllum salsugineum is a close relative. The light-compensation point and the photon irradiance required to saturate photosynthetic oxygen production were exponentially dependent on water temperature. Over the temperature range 15–35°C the light-compensation point increased from 2.4 to 16.9 μmol (PAR) m⁻² s⁻¹ for oxygen production while saturation photon irradiance increased from 41.5 to 138 μmol (PAR) m⁻² s⁻¹ for oxygen production and from 42.0 to 174 μmol (PAR) m⁻² s⁻¹ for CO₂ consumption. Respiration rates increased from 27.1 to 112.3 μmol (oxygen consumed) g⁻¹ dry weight h⁻¹ as temperature was increased from 15 to 35°C. The optimum temperature for productivity is 30°C.     

Studies on composition and stability of a large membered bacterial consortium degrading phenol

A ten member microbial consortium (AS) consisting of eight phenol-degrading and two non-phenol-degrading strains of bacteria was developed and maintained in a fed-batch reactor by feeding 500 mg l⁻¹ phenol for four years at 28 ± 3 °C. The consortium could degrade 99% of 500 mg l⁻¹ phenol after 24 hours incubation with a biomass increase of 2.6 × 10⁷ to 4 × 10¹² CFU ml⁻¹. Characterization of the members revealed that it consisted of 4 principal genera, Bacillus, Pseudomonas, Rhodococcus, Streptomyces and an unidentified bacterium. Phenol degradation by the mixed culture and Bacillus subtilis, an isolate from the consortium was compared using a range of phenol concentrations (400 to 700 mg l⁻¹) and by mixing with either 160 mg l⁻¹ glucose or 50 mg l⁻¹ of 2,4-dichlorophenol in the medium. Simultaneous utilization of unrelated mixed substrates (glucose/2,4-dichlorophenol) by the consortium and Bacillus subtilis, indicated the diauxic growth pattern of the organisms. A unique characteristic of the members of the consortia was their ability to oxidize chloro aromatic compounds via meta pathway and methyl aromatic compounds via ortho cleavage pathway. The ability of a large membered microbial consortia to maintain its stability with respect to its composition and effectiveness in phenol degradation indicated its suitability for bioremediation applications.     

Acid-base and metal ion-binding properties of flavin mononucleotide (FMN). Is a ‘dielectric’ effect responsible for the increased complex stability?

Due to contradictions in the literature we have redetermined the acid-base properties of riboflavin (=RiFl; vitamin B₂), i.e. 7,8-dimethyl-10-ribityl-isoalloxazine, and of flavin mononucleotide (FMN²⁻), also known as riboflavin 5′-phosphate, via potentiometric pH titrations (I = 0.1 M, NaNO₃; 25 °C). In contrast to various claims, the isoalloxazine ring cannot be protonated at pH > 1, a result in agreement with an early study (pK_a = −0.2; L. Michaelis, M.P. Schubert and C.V. Smythe, J. Biol. Chem., 116 (1936) 587–607); deprotonation of the ring system occurs in both compounds with pK_a 10. The pK_a value of 0.7 determined for the deprotonation of H₂(FMN) must be attributed to the release of the first proton from the fully protonated phosphate group; its second proton is released with pK_a = 6.18 in agreement with the acidity constants of various other monoprotonated monophosphate esters. The stability constants of the 1:1 complexes formed between Mg²⁺, Ca²⁺, Sr²⁺, Ba²⁺, Mn²⁺, Co²⁺, Ni²⁺, Cu²⁺, Zn²⁺ or Cd²⁺ (---M²⁺) and FMN²⁻ were determined by potentiometric pH titrations in aqueous solution (I = 0.1 M, NaNO₃; 25 °C). The log stability constants of all these M(FMN) complexes are about 0.2 log units higher than expected from the basicity of the phosphate group. This slight stability increase cannot be attributed to the formation of a seven-membered chelate involving the ribit-hydroxy group at C-4′ as the stability constants for the M²⁺ 1:1 complexes of glycerol 1-phosphate (G1P²⁻) demonstrate: G1P²⁻ contains the same structural unit which would also allow in this case the formation of the mentioned seven-membered chelate; however, the stability of the M(G1P) complexes is solely determined by the basicity of the phosphate group. Hence, in agreement with earlier conclusions (J. Bidwell, J. Thomas and J. Stuehr, J. Am. Chem. Soc., 108 (1986) 820–825) regarding Ni(FMN) one must conclude that the slight stability increase of the M(FMN) complexes has to be attributed to the isoalloxazine ring. The equality of the stability increase of the complexes for all the mentioned ten metal ions precludes its attribution to an interaction with an N site and makes a specific interaction with an O site also somewhat unlikely. In addition, carbonyl oxygens appear as not very favorable for the formation of macrochelates by a further interaction with already phosphate-coordinated metal ions. Therefore, we propose that the slight but significant stability increase originates from M(FMN) species (with a formation degree of about 30%) in which the hydrophobic flavin residue is close to the metal ion, thereby lowering the ‘effective’ dielectric constant in the microenvironment of the metal ion and thus indirectly promoting the −PO₃²⁻/M²⁺ interaction.     

Effect of low temperature (−30 to −60°C) on the reoxidation of the Photosystem II primary electron acceptor in the presence and absence of 3(3,4-dichlorophenyl)-1,1-dimethylurea

The fluorescence yield has been measured on spinach chloroplasts at low temperature (−30 to −60°C) for various dark times following a short saturating flash. A decrease in the fluorescence yield linked to the reoxidation of the Photosystem II electron acceptor Q is still observed at −60°C. Two reactions participate in this reoxidation: a back reaction or charge recombination and the transfer of an electron from Q⁻ to Pool A. The relative competition between these two reactions at low temperature depends upon the oxidation state of the donor side of the Photosystem II center:

1. (1) In dark-adapted chloroplasts (i.e. in States S₀+S₁ according to Kok, B., Forbush, B. and McGloin, M. (1970) Photochem. Photobiol. 11, 457–475), Q, reduced by a flash at low temperature, is reoxidized by a secondary acceptor and the positive charge is stabilized on the Photosystem II donor Z. Although this reaction is strongly temperature dependent, it still occurs very slowly at −60°C.

2. (2) When chloroplasts are placed in the S₂+S₃ states by a two-flash preillumination at room temperature, the reoxidation of Q⁻ after a flash at low temperature is mainly due to a temperature-independent back reaction which occurs with non-exponential kinetics.

3. (3) Long continuous illumination of a frozen sample at −30°C causes 6–7 reducing equivalents to be transferred to the pool. Thus, a sufficient number of oxidizing equivalents should have been generated to produce at least one O₂ molecule.

4. (4) A study of the back reaction in the presence of 3(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) shows the superposition of two distinct non-exponential reactions one temperature dependent, the other temperature independent.

Rapid scavenging of peroxynitrous acid by monohydroascorbate

The reaction of peroxynitrous acid with monohydroascorbate, over the concentration range of 250 μM to 50 mM of monohydroascorbate at pH 5.8 and at 25°C, was reinvestigated and the rate constant of the reaction found to be much higher than reported earlier (Bartlett, D.; Church, D. F.; Bounds, P. L.; Koppenol, W. H. The kinetics of oxidation of L-ascorbic acid by peroxynitrite. Free Radic. Biol. Med. 18:85–92; 1995; Squadrito, G. L.; Jin, X.; Pryor, W. A. Stopped-flow kinetics of the reaction of ascorbic acid with peroxynitrite. Arch. Biochem. Biophys. 322:53–59; 1995). The new rate constants at pH 5.8 are k₁ = 1 × 10⁶ M⁻¹ s⁻¹ and k₋₁ = 500 s⁻¹ for 25°C and k₁ = 1.5 × 10⁶ M⁻¹ s⁻¹ and k₋₁ = 1 × 10³ s⁻¹ for 37°C. These values indicate that even at low monohydroascorbate concentrations most of peroxynitrous acid forms an adduct with this antioxidant. The mechanism of the reaction involves formation of an intermediate, which decays to a second intermediate with an absorption maximum at 345 nm. At low monohydroascorbate concentrations, the second intermediate decays to nitrate and monohydroascorbate, while at monohydroascorbate concentrations greater than 4 mM, this second intermediate reacts with a second monohydroascorbate to form nitrite, dehydroascorbate, and monohydroascorbate. EPR experiments indicate that the yield of the ascorbyl radical is 0.24% relative to the initial peroxynitrous acid concentration, and that this small amount of ascorbyl radicals is formed concomitantly with the decrease of the absorption at 345 nm. Thus, the ascorbyl radical is not a primary reaction product. Under the conditions of these experiments, no homolysis of peroxynitrous acid to nitrogen dioxide and hydroxyl radical was observed. Aside from monohydroascorbate's ability to “repair” oxidatively modified biomolecules, it may play a role as scavenger of peroxynitrous acid.     

Ferrocenyl-derived face-capping ligands: syntheses and molecular structures of [(FcTt)Cu]4 and [FcTt]2M (M = Fe, Co, Ni; FcTt = ferrocenyltris((methylthio)methyl)borate)

The synthesis and characterization of a ferrocenyl-derived tridentate ligand, ferrocenyltris((methylthio)methyl)borate (FcTtP⁻), and its representative metal complexes, [(FcTt)Cu]₄ and [FcTt]₂M (M = Fe, Co and Ni), are reported. The M = Fe complex exhibits spin-crossover behavior with a μ_eff = 1.19 μ_B at 25°C. The low-spin Co(II) derivative (1.88 μ_B) exhibits a characteristic axial electron paramagnetic resonance (EPR) spectrum, g_av = 2.13, A = 53 G and A_¦ = 43 G. The [FcTt]₂M complexes display reversible two-electron redox processes assigned to ligand-centered events about 200 mV negative of the ferrocene-ferrocenium couple. [(FcTt)Cu]₄ and [FcTt]₂Ni have been characterized by X-ray diffraction. X-ray data for [(FcTt)Cu]₄: monoclinic space group C2/c, with a = 24.3747(3) Å, b = 20.0857(2) Å, c = 17.2747(4) Å, β = 95.843(1)°, V = 8413.5(3) ų, and Z = 4; [FcTt]₂Ni: monoclinic space group C2/c, with a = 12.6220(3) Å, b = 11.6002(3) Å, c = 25.0125(7) Å, β = 94.067(1)°, V = 3653.1(2) ų, and Z = 4.     

Effects of elevated water temperature on the critical swim speeds of yearling rainbow trout Salmo gairdneri

Effects of elevated water temperature on the critical swim speeds of rainbow trout, Salmo gairdneri, were investigated. Trout acclimated to 10°C were exposed to 10, 15, 20 and 20°C while swimming and at rest. Initial swim speed of 20 cms⁻¹ was increased in 10 cms⁻¹ increments every 20 min until the fish fatigued. Critical swim speeds were calculated in absolute values (cms⁻¹) and relative performance values (body lengths s⁻¹). Critical swim speeds were similar at 10, 15 and 25°C. Swimming performance was significantly decreased at 25°C. Performance measured as critical swim speed was unaffected by temperature elevations up to 10°C above acclimation temperature of 10°C.     

Glutathione-proline activation of feeding behavior in the zoanthid Palythoa psammophilia Walsh & Bowers

Palythoa psammophilia Walsh & Bowers has a well coordinated, stereotyped feeding response, the culminating step of which is ingestion; this may be elicited by the synergistic effect of the tripeptide glutathione and the -imino acid, proline. Either activator acting separately causes responses only at high concentrations (above 10⁻⁵ M for glutathione; above 10⁻⁴ M for proline) in a reduced number of animals and at a low rate (5.00 ± 1.73 min in 5 × 10⁻³ M solutions of glutathione; 11.10±3.74 min in 5 × 10⁻³ M solutions of proline). Highest percentages of response were obtained in combinations where glutathione was at a concentration of 5 × 10⁻³ M and proline at 5 × 10⁻⁴ M or in combinations of glutathione at concentrations 5 × 10⁻⁶ M and proline at 5 × 10⁻⁵ M. The speed of ingestion is considerably enhanced when these activators are combined (1.17±1.18 min).