High-resolution,High-speed,Three-dimensional Video Imaging with Digital Fringe Projection Techniques

Digital fringe projection (DFP) techniques provide dense 3D measurements of dynamically changing surfaces. Like the human eyes and brain, DFP uses triangulation between matching points in two views of the same scene at different angles to compute depth. However, unlike a stereo-based method, DFP uses a digital video projector to replace one of the cameras¹. The projector rapidly projects a known sinusoidal pattern onto the subject, and the surface of the subject distorts these patterns in the camera’s field of view. Three distorted patterns (fringe images) from the camera can be used to compute the depth using triangulation.Unlike other 3D measurement methods, DFP techniques lead to systems that tend to be faster, lower in equipment cost, more flexible, and easier to develop. DFP systems can also achieve the same measurement resolution as the camera. For this reason, DFP and other digital structured light techniques have recently been the focus of intense research (as summarized in^1-5). Taking advantage of DFP, the graphics processing unit, and optimized algorithms, we have developed a system capable of 30 Hz 3D video data acquisition, reconstruction, and display for over 300,000 measurement points per frame^6,7. Binary defocusing DFP methods can achieve even greater speeds⁸.Diverse applications can benefit from DFP techniques. Our collaborators have used our systems for facial function analysis⁹, facial animation¹⁰, cardiac mechanics studies¹¹, and fluid surface measurements, but many other potential applications exist. This video will teach the fundamentals of DFP techniques and illustrate the design and operation of a binary defocusing DFP system.     

Scutellaria baicalensis extract and baicalein inhibit replication of SARS-CoV-2 and its 3C-like protease in vitro

COVID-19 has become a global pandemic and there is an urgent call for developing drugs against the virus (SARS-CoV-2). The 3C-like protease (3CL^pro) of SARS-CoV-2 is a preferred target for broad spectrum anti-coronavirus drug discovery. We studied the anti-SARS-CoV-2 activity of S. baicalensis and its ingredients. We found that the ethanol extract of S. baicalensis and its major component, baicalein, inhibit SARS-CoV-2 3CL^pro activity in vitro with IC₅₀’s of 8.52 µg/ml and 0.39 µM, respectively. Both of them inhibit the replication of SARS-CoV-2 in Vero cells with EC₅₀’s of 0.74 µg/ml and 2.9 µM, respectively. While baicalein is mainly active at the viral post-entry stage, the ethanol extract also inhibits viral entry. We further identified four baicalein analogues from other herbs that inhibit SARS-CoV-2 3CL^pro activity at µM concentration. All the active compounds and the S. baicalensis extract also inhibit the SARS-CoV 3CL^pro, demonstrating their potential as broad-spectrum anti-coronavirus drugs.     

Conformation dependent binding of netropsin and distamycin to DNA and DNA model polymers

The binding of the antibiotics netropsin and distamycin A to DNA has been studied by thermal melting, CD and sedimentation analysis. Netropsin binds strongly at antibiotic/nucleotide ratios up to at least 0.05. CD spectra obtained using DNA model polymers reveal that netropsin binds tightly to poly (dA) · poly (dT), poly (dA-dT) · poly(dA-dT) and poly (dI-dC) · poly (dI-dC) but poorly, if at all, to poly (dG) · poly (dC). Binding curves obtained with calf thymus DNA reveal one netropsin-binding site per 6.0 nucleotides (K_a=2.9 · 10⁵ M⁻¹); corresponding values for distamycin A are one site per 6.1 nucleotides with K_a= 11.6 · 10⁵ M⁻¹. Binding sites apparently involve predominantly A·T-rich sequences whose specific conformation determines their high affinity for the two antibiotics. It is suggested that the binding is stabilized primarily by hydrogen bonding and electrostatic interactions probably in the narrow groove of the DNA helix, but without intercalation. Any local structural deformation of the helix does not involve unwinding greater than approximately 3° per bound antibiotic molecule.     

Acetylcholine Receptors : Revised Estimates of Extrajunctional Receptor Density in Denervated Rat Diaphragm

The number of extrajunctional acetylcholine receptors (¹²⁵I-labeled α-bungarotoxin binding sites) per unit length of muscle fiber and the average fiber circumference were determined for rat diaphragm muscle fibers denervated 0, 2, 4, 7, 10, and 14 days. From these data receptor densities (sites per square micrometer of surface) were calculated. Values thus obtained were considerably lower than those estimated previously by autoradiography. Receptor density increased from < 6 sites/µm² in innervated muscle to 635 ± 29 sites/µm² 14 days after denervation. The form of the relationship between receptor density and acetylcholine sensitivity and the time-course of change in receptor density after denervation are as previously reported.     

A study of some factors that influence the iodination of ox insulin

1. The influence of carrier iodide, iodine monochloride and pH on the labelling of ox insulin with ¹³¹I by the iodine monochloride method have been studied. 2. The quantitative effect of the iodide in the radioactive iodine preparation was that predicted from a calculation of its specific activity. No other interfering factors were detected in the [¹³¹I]iodide solutions used. 3. Increasing the molar ratio of iodine monochloride to insulin resulted in an increase followed progressively by a decrease in the proportion of ¹³¹I bound, while the total iodine bound increased to an amount characteristic of pH and thereafter remained constant. 4. The influence of pH on the iodination of insulin with iodine monochloride was complex and the pH curve showed two maxima, at pH2·8 and 6·4. At pH2·8 it was not possible to exceed 8 atoms of iodine bound per molecule by increasing the molar ratio of iodine monochloride. Similarly, at pH6·4 the substitution value of 11·5 atoms of iodine per molecule could not be exceeded. 5. Iodinated insulins containing an average of 1·96, 2·74, 6·0 and 7·0 atoms of iodine per molecule fully retained the ability to bind guinea-pig anti-(ox insulin) serum, and the ability to compete with unlabelled insulin for antibody sites only became significantly changed in the most highly substituted preparations and in the presence of large concentrations of unlabelled insulin. 6. The method for the iodination of insulin with 98% incorporation of ¹³¹I by using chloramine-t is described. 7. ¹³¹I-iodinated insulin prepared with graded quantities of chloramine-t in excess of that required for efficient labelling was less efficiently bound by guinea-pig anti-(ox insulin) serum than insulin labelled by the iodine monochloride method.     

Kinetics of thiamin cleavage by sulphite

Results are presented on the rate of thiamin cleavage by sulphite in aqueous solutions as affected by temperature (20–70°), pH(2·5–7·0), and variation of the concentration of either thiamin (1–20μm) or sulphite (10–5000μm as sulphur dioxide). Plots of the logarithm of percentage of residual thiamin against time were found to be linear and cleavage thus was first-order with respect to thiamin. At pH5 the rate was also found to be proportional to the sulphite concentration. In the pH region 2·5–7·0 at 25° the rate constant was 50m⁻¹hr.⁻¹ at pH5·5–6·0, and decreased at higher or lower pH values. The rate of reaction increased between 20° and 70°, indicating a heat of activation of 13·6kcal./mole.     

Neonicotinoid Binding,Toxicity and Expression of Nicotinic Acetylcholine Receptor Subunits in the Aphid Acyrthosiphon pisum

Neonicotinoid insecticides act on nicotinic acetylcholine receptor and are particularly effective against sucking pests. They are widely used in crops protection to fight against aphids, which cause severe damage. In the present study we evaluated the susceptibility of the pea aphid Acyrthosiphon pisum to the commonly used neonicotinoid insecticides imidacloprid (IMI), thiamethoxam (TMX) and clothianidin (CLT). Binding studies on aphid membrane preparations revealed the existence of high and low-affinity binding sites for [³H]-IMI (Kd of 0.16±0.04 nM and 41.7±5.9 nM) and for the nicotinic antagonist [¹²⁵I]-α-bungarotoxin (Kd of 0.008±0.002 nM and 1.135±0.213 nM). Competitive binding experiments demonstrated that TMX displayed a higher affinity than IMI for [¹²⁵I]-α-bungarotoxin binding sites while CLT affinity was similar for both [¹²⁵I]-α-bungarotoxin and [³H]-IMI binding sites. Interestingly, toxicological studies revealed that at 48 h, IMI (LC₅₀ = 0.038 µg/ml) and TMX (LC₅₀ = 0.034 µg/ml) were more toxic than CLT (LC₅₀ = 0.118 µg/ml). The effect of TMX could be associated to its metabolite CLT as demonstrated by HPLC/MS analysis. In addition, we found that aphid larvae treated either with IMI, TMX or CLT showed a strong variation of nAChR subunit expression. Using semi-quantitative PCR experiments, we detected for all insecticides an increase of Apisumα10 and Apisumβ1 expressions levels, whereas Apisumβ2 expression decreased. Moreover, some other receptor subunits seemed to be differently regulated according to the insecticide used. Finally, we also demonstrated that nAChR subunit expression differed during pea aphid development. Altogether these results highlight species specificity that should be taken into account in pest management strategies.     

The binding of calcium and yttrium ions to a glycoprotein from bovine cortical bone

The binding of Ca²⁺ and Y³⁺ to an acidic glycoprotein from bovine cortical bone, bone sialoprotein, was determined from the titration curves at I 0·2 in the presence and absence of the cations. The binding of Y³⁺ was greater than that of Ca²⁺. The value for the association constant, k, for the interaction with Y³⁺ increased with pH, from log k 2·93 at pH3·4 to log k 3·50 at pH4·4, and the number of binding sites/mol. increased from 4·6 at pH3·4 to 9·1 at pH4·4. It is proposed that the binding site consists of three carboxyl groups, but it is likely that the binding is a strong electrostatic interaction rather than a co-ordination linkage. A chondroitin sulphate–protein complex also extracted from bovine cortical bone interacted with Y³⁺ and Ca²⁺ to a similar extent as did bone sialoprotein. It is suggested that these materials are present in bone at the resting and resorbing surfaces and that they contribute to the deposition of yttrium, americium and plutonium at these sites.     

Effects of Temperature and Pressure on Sulfate Reduction and Anaerobic Oxidation of Methane in Hydrothermal Sediments of Guaymas Basin

Rates of sulfate reduction (SR) and anaerobic oxidation of methane (AOM) in hydrothermal deep-sea sediments from Guaymas Basin were measured at temperatures of 5 to 200°C and pressures of 1 × 10⁵, 2.2 × 10⁷, and 4.5 × 10⁷ Pa. A maximum SR of several micromoles per cubic centimeter per day was found at between 60 and 95°C and 2.2 × 10⁷ and 4.5 × 10⁷ Pa. Maximal AOM was observed at 35 to 90°C but generally accounted for less than 5% of SR.     

Alona iheringula Sinev & Kotov, 2004 (Crustacea,Anomopoda, Chydoridae,Aloninae): Life Cycle and DNA Barcode with Implications for the Taxonomy of the Aloninae Subfamily

Knowledge of reproductive rates and life cycle of the Cladocera species is essential for population dynamic studies, secondary production and food webs, as well as the management and preservation of aquatic ecosystems. The present study aimed to understand the life cycle and growth of Alona iheringula Kotov & Sinev, 2004 (Crustacea, Anomopoda, Chydoridae), a Neotropical species, as well as its DNA barcoding, providing new information on the Aloninae taxonomy. The specimens were collected in the dammed portion of the Cabo Verde River (21°26′05″ S and 46°10′57″ W), in the Furnas Reservoir, Minas Gerais State, Brazil. Forty neonates were observed individually two or three times a day under controlled temperature (25±1°C), photoperiod (12 h light/12 h dark) and feeding (Pseudokirchneriella subcapitata at a concentration of 10⁵ cells.mL⁻¹ and a mixed suspension of yeast and fish feed in equal proportion). Individual body growth was measured daily under optical microscope using a micrometric grid and 40× magnification. The species had a mean size of 413(±29) µm, a maximum size of 510 µm and reached maturity at 3.24(±0.69) days of age. Mean fecundity was 2 eggs per female per brood and the mean number of eggs produced per female during the entire life cycle was 47.6(±6.3) eggs per female. The embryonic development time was 1.79(±0.23) days and the maximum longevity was 54 days. The species had eight instars throughout its life cycle and four instars between neonate and primipara stage. The present study using molecular data (a 461 bp smaller COI fragment) demonstrated a deep divergence in the Aloninae subfamily.     

A METHOD FOR THE DETERMINATION OF DIFFUSION CONSTANTS AND THE CALCULATION OF THE RADIUS AND WEIGHT OF THE HEMOGLOBIN MOLECULE

A method is described for determining the diffusion coefficient of solutes by determining the rate of passage of the solute through a thin porous membrane between two solutions of different concentration. The method has been used to determine the diffusion coefficient of carbon monoxide hemoglobin. This was found to be 0.0420 ± 0.0005 cm.² per day at 5°C. The molecular weight of carbon monoxide hemoglobin calculated by means of Einstein''s equation from this quantity is 68,600 ± 1,000.     

EFFECT OF THE NUMBER OF SPRINT REPETITIONS ON THE VARIATION OF BLOOD LACTATE CONCENTRATION IN REPEATED SPRINT SESSIONS

The aim of the present study was to examine the effect of number of sprint repetitions on the variation of blood lactate concentration (blood [La]) during different repeated-sprint sessions in order to find the appropriate number of sprint repetitions that properly simulates the physiological demands of team sport competitions. Twenty male team-sport players (age, 22.2 ± 2.9 years) performed several repeated-sprint sessions (RSS) consisting of 1, 2, 3, 4, 5, 9, or 10 repetitions of 30 m shuttle sprints (2 × 15 m) with 30 s recovery in between. The blood [La] was obtained after 3 min of recovery at the end of each RSS. The present study showed that for RSS of 3 sprints (RSS3) there was a high increase (p<0.001) in blood [La], which reached approximately fivefold resting values (9.4±1.7 mmol · l⁻¹) and then remained unchanged for the RSS of 4 and 5 sprints (9.6±1.4 and 10.5±1.9 mmol · l⁻¹, p=0.96 and 0.26, respectively). After RSS9 and RSS10 blood [La] further significantly increased to 12.6 and 12.7 mmol · l⁻¹, p<0.001, respectively. No significant difference was found between RSS3, RSS4 and RSS5 for the percentage of sprint speed decrement (Sdec) (1.5±1.2; 2.0±1.1 and 2.6±1.4%, respectively). There was also no significant difference between RSS9 and RSS10 for Sdec (3.9±1.3% and 4.5±1.4%, respectively). In conclusion, the repeated-sprint protocol composed of 5 shuttle sprint repetitions is more representative of the blood lactate demands of the team sports game intensity.     

Cyclic variations in nitrogen uptake rate in soybean plants

Uptake of NO₃⁻ by nonnodulated soybean plants (Glycine max L. Merr. cv Ransom) growing in flowing hydroponic culture at 22 and 14°C root temperatures was measured daily during a 31-day growth period. Ion chromatography was used to determine removal of NO₃⁻ from solution during each 24-hour period. At both root-zone temperatures, rate of NO₃⁻ uptake per plant oscillated with a periodicity of 3 to 5 days. The rate of NO₃⁻ uptake per plant was consistently lower at 14°C than 22°C. The lower rate of NO₃⁻ uptake at 14°C during the initial 5 to 10 days was caused by reduced uptake rates per gram root dry weight, but with time uptake rates per gram root became equal at 14 and 22°C. Thereafter, the continued reduction in rate of NO₃⁻ uptake per plant at 14°C was attributable to slower root growth.     

Ultrasound-mediated DNA transfer for bacteria

In environmental microbiology, the most commonly used methods of bacterial DNA transfer are conjugation and electroporation. However, conjugation requires physical contact and cell–pilus–cell interactions; electroporation requires low-ionic strength medium and high voltage. These limitations have hampered broad applications of bacterial DNA delivery. We have employed a standard low frequency 40 kHz ultrasound bath to successfully transfer plasmid pBBR1MCS2 into Pseudomonas putida UWC1, Escherichia coli DH5α and Pseudomonas fluorescens SBW25 with high efficiency. Under optimal conditions: ultrasound exposure time of 10 s, 50 mM CaCl₂, temperature of 22°C, plasmid concentration of 0.8 ng/µl, P. putida UWC1 cell concentration of 2.5 × 10⁹ CFU (colony forming unit)/ml and reaction volume of 500 µl, the efficiency of ultrasound DNA delivery (UDD) was 9.8 ± 2.3 × 10⁻⁶ transformants per cell, which was nine times more efficient than conjugation, and even four times greater than electroporation. We have also transferred pBBR1MCS2 into E. coli DH5α and P. fluorescens SBW25 with efficiencies of 1.16 ± 0.13 × 10⁻⁶ and 4.33 ± 0.78 × 10⁻⁶ transformants per cell, respectively. Low frequency UDD can be readily scaled up, allowing for the application of UDD not only in laboratory conditions but also on an industrial scale.     

Estimation of the fructose diphosphatase-phosphofructokinase substrate cycle in the flight muscle of Bombus affinis

1. Substrate cycling of fructose 6-phosphate through reactions catalysed by phosphofructokinase and fructose diphosphatase was estimated in bumble-bee (Bombus affinis) flight muscle in vivo. 2. Estimations of substrate cycling of fructose 6-phosphate and of glycolysis were made from the equilibrium value of the ³H/¹⁴C ratio in glucose 6-phosphate as well as the rate of ³H release to water after the metabolism of [5-³H,U-¹⁴C]glucose. 3. In flight, the metabolism of glucose proceeded exclusively through glycolysis (20.4μmol/min per g fresh wt.) and there was no evidence for substrate cycling. 4. In the resting bumble-bee exposed to low temperatures (5°C), the pattern of glucose metabolism in the flight muscle was altered so that substrate cycling was high (10.4μmol/min per g fresh wt.) and glycolysis was decreased (5.8μmol/min per g fresh wt.). 5. The rate of substrate cycling in the resting bumble-bee flight muscle was inversely related to the ambient temperature, since at 27°, 21° and 5°C the rates of substrate cycling were 0, 0.48 and 10.4μmol/min per g fresh wt. respectively. 6. Calcium ions inhibited fructose diphosphatase of the bumble-bee flight muscle at concentrations that were without effect on phosphofructokinase. The inhibition was reversed by the presence of a Ca²⁺-chelating compound. It is proposed that the rate of fructose 6-phosphate substrate cycling could be regulated by changes in the sarcoplasmic Ca²⁺ concentration associated with the contractile process.     

Useful Host-Vector Systems in Bacillus stearothermophilus

We isolated a highly transformable thermophile, Bacillus stearothermophilus SIC1, which exhibited the following features. The growth temperature ranged from 45 to 65°C in L broth. The maximum cell concentration in 2L broth (2% tryptone, 1% yeast extract, 0.5% NaCl, pH 7.2) was determined as an optical density at 660 nm of 7.8, and the generation time was 11 min at 60°C. Strain SIC1 was a prototroph and was transformed by the protoplast procedure not only with repB plasmids (high-copy-number plasmids such as pTB913 and pUB110) but also with repA plasmids (low-copy-number plasmids such as pTB53). Transformation efficiencies with repB and repA plasmids were about 2 × 10⁶ to 5 × 10⁶ and 5 × 10⁴ transformants per μg of DNA, respectively. The transformant carrying plasmid pTB913Y/K could grow at 63°C in the presence of kanamycin. The regeneration frequency of protoplasts was 60%, and only 1 day was needed for regeneration at 55°C.     

Electrical Properties and Acetylcholine Sensitivity of Singly and Multiply Innervated Avian Muscle Fibers

Membrane constants and distribution of acetylcholine (ACh) receptors were determined for multiply innervated fibers of the anterior latissimus dorsi (ALD) and singly innervated fibers of the posterior latissimus dorsi (PLD) muscles of 3–6 month old chickens. The values of the various membrane constants were: length constant, 1.78 mm (mean) in ALD, 0.68 mm in PLD; time constant, 35 msec in ALD, 3.7 msec in PLD; transverse membrane resistance, 4388 Ω cm² in ALD, 561 Ω cm² in PLD; and membrane capacitance, 8.2 µF/cm² in ALD, 7.0 µF/cm² in PLD. Peaks of ACh sensitivity occurred at intervals of ca. 740 µ on ALD fibers with a low sensitivity remaining between peaks. Only one peak of ACh sensitivity was detected on PLD fibers. The maximum ACh sensitivity found was 5 ± 4 mv/ncoul for fibers of the ALD and 77 ± 60 mv/ncoul for fibers of the PLD. The distance over which this sensitivity fell to 0.1 was ca. 225 µ in the ALD and 140 µ in the PLD. The membranes of these two muscle fiber types differ widely regarding some electrical properties and the disposition of ACh-sensitive receptor sites.     

Ageing and the influx of water into radish root-hair cells

A micropotometric device previously described by the writer was used to determine quantitatively the velocity of water influx in cubic microns per square micron of hair surface per minute of comparatively older and younger root hairs of radish seedlings in a humid atmosphere at 29° ± 1°C. when the micropotometers were filled with Hoagland solution at pH 6.8. In each experiment, measurements were made on two hairs of different length and different age on a given root and the hairs were inserted into the micropotometers a sufficient distance so that the area of immersion, 13,200µ², was the same in each instance. The range of velocities of water influx through the immersed surface was 4.46 to 1.16µ³/µ²/min. for the younger and shorter hairs which varied in length from 280 to 460 microns. The range of velocities of water influx through the immersed surface of the older hairs which varied from 661 to 2300 microns in length was 1.94 to 0.47µ³/µ²/min. The data indicate that water entry slows down in older hairs independent of root length. Estimations were made of the times to replace hair volumes based upon the mean velocities of water entry of the immersed areas. It was found that the time for the hairs to absorb an amount of water equivalent to their own volumes under the conditions specified was a matter of minutes or less; the range was 0.90 to 8.51 minutes.     

STEREOLOGICAL ANALYSIS OF MAMMALIAN SKELETAL MUSCLE : I. Soleus Muscle of the Adult Guinea Pig

A quantitative analysis of the volumes, surface areas, and dimensions of the ultrastructural components in the soleus muscle fibers of the guinea pig was made by using point counting methods of stereology. Muscle fibers have structural orientation (anisotropy) and have spatial gradients of the structures within the fiber; therefore the standard stereological methods were modified where necessary. The entire analysis was repeated at two section orientations to test the modifications and identical results obtained from both. The volume of lipid droplets was 0.20 ± 0.06% (mean ± standard error, n = 5 animals) and the nuclei volume was 0.86 ± 0.20% of the fiber volume. The total mitochondrial volume was 4.85 ± 0.66% of the fiber volume with about one-third being found in an annulus within 1 µm of the sarcolemma. The mitochondrial volume in the remaining core of the fiber was 3.6 ± 0.4%. The T system has a volume of 0.14 ± 0.01% and a surface area of 0.064 ± 0.005 µm²/µm³ of the fiber volume. The surface area of the sarcolemma is 0.116 ± 0.013 µm²/µm³ which is twice the T system surface area. The volume of the entire sarcoplasmic reticulum is 3.52 ± 0.33% and the surface area is 0.97 ± 0.09 µm²/µm³. The sarcoplasmic reticulum is composed of the terminal cisternae whose volume is 1.04 ± 0.19% and surface area is 0.24 ± 0.05 µm²/µm³. The tubules of the sarcoplasmic reticulum in the I band and A band have volumes of 1.97 ± 0.24% and 0.51 ± 0.08%, and the surface areas of the I and A band reticulum are 0.56 ± 0.07 µm²/µm³ and 0.16 ± 0.04 µm²/µm³, respectively. The Z line width, myofibril and fiber diameters were measured.     

THE INFLUENCE OF ENVIRONMENTAL TEMPERATURE ON THE UTILIZATION OF FOOD ENERGY IN BABY CHICKS

1. An optimum of environmental temperature is to be expected for the utilization of food energy in warm blooded animals if their food intake is determined by their appetite. 2. Baby chicks were kept in groups of five chicks in a climatic cabinet at environmental temperatures of 21°, 27°, 32°, 38°, and 40°C. during the period of 6 to 15 days of age. The intake of qualitatively complete food was determined by their appetite. Food intake, excretion, and respiratory exchange were measured. Control chicks from the same hatch as the experimental groups were raised in a brooder and were given the same food as the experimental chicks. The basal metabolism of each experimental group was determined from 24 to 36 hours without food at the age of 16 days. 3. The daily rate of growth increased with decreasing environmental temperature from 2.74 gm. at 40°C. to 4.88 gm. at 21°C. This was 4.2 to 6.5 per cent of their body weight. 4. The amount of food consumed increased in proportion to the decrease in temperature. 5. The availability of the food, used for birds instead of the digestibility and defined as See PDF for Structure showed an optimum at 38°C. 6. The CO₂ production increased from 2.95 liters CO₂ per day per chick at 40°C. to 6.25 liters at 21°C. Per unit of the 3/4 power of the body weight, 23.0 liters CO₂ per kilo^3/4 was produced at 40°C. and 43.4 liters per kilo^3/4 at 21°C. The CO₂ production per unit of 3/4 power of the weight increased at an average rate of approximately 1 per cent per day increase in age. The R.Q. was, on the average, 1.04 during the day and 0.92 during the night. 7. The net energy is calculated on the basis of C and N balances. A maximum of 11.8 Cal. net energy per chick per day was found at 32°C. At 21°C. only 6.9 Cal. net per day per chick was produced and at 40°C. an average of 6.7 Cal. 8. The composition of the gained body substance changed according to the environmental temperature. The protein stored per gram increase in body weight varied from 0.217 to 0.266 gm. protein and seemed unrelated to the temperature. The amount of fat per gram gain in weight dropped from a maximum of 0.153 gm. at 32°C. to 0.012 gm. at 21°C. and an average of 0.107 gm. at 40°C. The energy content per gram of gain in weight had its maximum of 2.95 Cal. per gm. at 38°C. and its minimum of 1.41 Cal. per gm. at 21°C. at which temperature the largest amount of water (0.763 gm. per gm. increase in body weight) was stored. 9. The basal metabolism increased from an average of 60 Cal. per kilo^3/4 at an environmental temperature of 40°C. to 128 Cal. per kilo^3/4 at 21°C. No indication of a critical temperature was found. 10. The partial efficiency, i.e. the increase in net energy per unit of the corresponding increase in food energy, seemed dependent on the environmental temperature, reaching a maximum of 72 per cent of the available energy at 38°C. and decreasing to 57 per cent at 21°C. and to an average of 60 per cent at 40°C. 11. The total efficiency, i.e. the total net energy produced per unit of food energy taken in, was maximum (34 per cent of the available energy) at 32°C., dropped to 16 per cent at 21°C., and to an average of 29 per cent at 40°C.