Oxygen-uptake in snakes - is there a reduction in fossorial species

Oxygen uptake of the fossorial blind snake (Typhlops reticulatus) and the semifossorial coral snake (Micrurus ibiboboca) was measured at 20 and 30 degrees C. Oxygen uptake of blind snakes was within the normal range, whereas oxygen uptake of coral snakes was in the lower end of values reported for snakes. The results do not support the hypothesis of reduced oxygen uptake in fossorial reptiles.

ONTOGENIC VARIATION OF OXYGEN-UPTAKE IN THE PITVIPER BOTHROPS MOOJENI (SERPENTES, VIPERIDAE)

The scaling of oxygen uptake was measured along the ontogeny, in the neotropical pitviper Bothrops moojeni. Allometric relationship between oxygen uptake and body mass, quantified for juveniles, sub-adults and adults, showed the same mass coefficient and exponent. The uniformity of mass constants along ontogeny suggests that B. moojeni is energetically homomorphic. Variation in mass seem to be the sole determinant of oxygen uptake, and structural modifications have no effect on the metabolic rate. Applications of the homomorphism principle to assess variations in mass coefficient and exponent for intraspecific analysis of metabolism in reptiles are discussed. B. moojeni had an oxygen consumption in the range reported for viperids, but lower than that for colubrid snakes of similar size. Possible causative reasons for this pattern is discussed.

LOW-PROTEIN DIET IMPAIRS GLUCOSE-INDUCED INSULIN-SECRETION FROM AND CA-45 UPTAKE BY PANCREATIC RAT ISLETS

Glucose-induced insulin secretion rom and Ca-45 uptake by isolated pancreatic islets, derived from rats fed with normal (NPD) or low protein diet (LPD), were studied. Insulin secretion from both types of islets in response to increasing concentrations of glucose followed an S-shaped pattern. However, basal secretion observed at substimulatory concentrations of glucose (0-5.6 mM), as well as maximal release, obtained at 16.7 mM or higher glucose concentrations were significantly reduced in islets from LPD. Furthermore, in LPD rat islets, the dose-response curve to glucose was clearly shifted to the right compared with NPD islets, with the half-maximal response occurring at 8.5 and 14.4 mM glucose for NPD and LPD islets, respectively. In islets from NPD rats, the Ca-45 content, after 5 or 90 min in the presence of 8.3 mM glucose, was higher than that observed for islets kept at 2.8 mM glucose and increased further at 16.7 mM glucose. After 5 min of incubation, the Ca-45 uptake by LPD islets in the presence of 8.3 mM glucose was slightly higher than basal values (2.8 mM glucose); however, no further increase in the Ca-45 uptake was noticed at 16.7 mM glucose. In LPD islets a significant increase in Ca-45 uptake over basal values was registered only at 16.7 mM glucose, after 90 min of incubation. These data indicate that the poor secretary response to glucose observed in islets from LPD rats may be related to a defect in the ability of glucose to increase Ca2+ uptake and/or to reduce Ca2+ efflux from beta-cells.

Yield and nitrogen uptake of peanuts as affected by lime, cobalt, and molybdenum

Peanut response to lime has been associated to calcium (Ca) nutrition, but a higher nitrogen (N) uptake has been observed in limed plots probably due to an increase in molybdenum (Mo) availability. A two-year experiment was conducted to study the effects of Mo, cobalt (Co), and liming on peanut yields and N nutrition. Peanut seeds were treated with Mo and/or Co and grown in soil with base saturation about 13, 41, 57, and 71%. There was no effect of seed treatment with Co on peanut yields or N nutrition. Liming and Mo application increased N contents in the leaves. Nitrogen uptake was increased by Mo and liming in cv. Tatu and only by liming in cv. Tupa. Manganese (Mn) contents in the leaves were decreased by liming. The higher yields were observed when the Ca/Mn ratio in the leaves was above 25. In acid soils, low availability of Mo and Mn toxicity can impair N acquisition by peanut plants and decrease grain yields.

Thiabendazole uptake and persistence in lemons following postharvest dips at 50 degrees C

Thiabendazole (TBZ) uptake and degradation rate in lemon fruits, following prestorage dipping at 50 degrees C in mixtures containing different amounts of fungicide, was compared with those measured after treatment at standard room temperature. TBZ residues were strictly dependent upon the amount of fungicide. Following 1,200 ppm TBZ dipping at 20 degrees C residue uptake in fruit was the same that would have been accumulated with ca. 150 ppm fungicide at 50 degrees C, a value that can be calculated due to the linear relationship between the residue in fruit after treatment and fungicide concentration TBZ residues showed great persistence during fruit storage: after 13 weeks at 8 degrees C and 1 week at 20 degrees C residues in fruit averaged ca. 70% of their initial values. In this study it was shown that it is possible to employ remarkably low amounts of TBZ (ca. 150 mg kg(-1)), when applied in combination with water at 50 degrees C, and keep the same residues of fruit treated at room temperature with high amounts of TBZ (1,200 mg kg(-1)).

CIRCADIAN OSCILLATORY PATTERNS OF OXYGEN-UPTAKE IN INDIVIDUAL WORKERS OF THE ANT CAMPONOTUS-RUFIPES

Respiratory rates of individual workers of Camponotus rufipes Fabricius (Hymenoptera: Formicidae) were measured at 25-degrees-C and LD 12:12 h (lights on 06.00 hours), DL 12:12 h (lights on 18.00 hours), LL (850 lux) and DD (red light, 20-30 lux), using the micro-Warburg technique. Worker ants were collected from natural nest during the winter of 1987 in a woodland park in the region of Rio Claro, São Paulo, Brazil. The respiration of ants showed a circadian rhythm with acrophase ranging from 20 h 41 min to 01 h 18 min and from 10 h 32 min to 12 h 22 min at LD and DD, respectively. In constant darkness the rhythmometric variables were similar to those presented by ants kept at LD 12:12 h. Under constant light no circadian rhythm in the respiration rates was found. A reduction in the amplitude was observed, indicating an inhibitory effect of this light regime on the respiration process.

Inhibition of hydrogen peroxide, nitric oxide and TNF-alpha production in peritoneal macrophages by ethyl acetate fraction from Alchornea glandulosa

The effects of Alchornea glandulosa ethyl acetate fraction (AGF) on hydrogen peroxide (H2O2), nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) production in peritoneal macrophages activated with lipopolysaccharide (LPS) or phorbol myristate acetate (PMA) were investigated. Analysis by thin layer chromatography (TLC) of AGF showed several constituents, including flavonoids, which may have anti-inflammatory activity. Inhibitory effects of the fraction in H2O2 and NO production ranged from 8.59 +/- 7.84% to 70.56 +/- 4.16% and from 16.06 +/- 3.65% to 38.73 +/- 3.90%, respectively. The TNF-alpha production was only partially inhibited in the tested concentrations (12.21 +/- 6.23%-15.16 +/- 0.96%). According to these results, it is suggested that AGF has anti-inflammatory activity. This medicinal plant may have therapeutic potential in the control of inflammatory disorders.

Interleukin-10 secreted by B-1 cells modulates the phagocytic activity of murine macrophages in vitro

As demonstrated previously in our laboratory, B-1 cells migrate from the peritoneal cavity of mice and home to a distant site of inflammation to become macrophage-like cells. However, the influence that these cells might have on the kinetics and fate of the inflammatory process is not known. Considering that macrophages are pivotal in the inflammatory reaction, we decided to investigate the possible influence B-1 cells could have on macrophage activities in vitro. Our results show that peritoneal macrophages from Xid mice, a mouse strain deprived of B-1 cells, have higher phagocytic indexes for zymozan particles when compared with macrophages from wild-type mice. Moreover, macrophages from wild-type mice have a lower ability to release nitric oxide and hydrogen peroxide when compared with macrophages from Xid mice. Experiments using cocultures of B-1 cells and macrophages from Xid mice in transwell plates demonstrated that B-1 cells down-regulate macrophage activities. These observations also indicate that this phenomenon is not due to a physical interaction between these two cell populations. As B-1 cells are one of the main sources of interleukin (IL)-10, we demonstrate in this study that adherent peritoneal cells from Xid mice produce significantly less amounts of this cytokine in culture when compared with IL-10 production by cells from wild-type mice. When B-1 cells from IL-10 knock-out mice and macrophages from wild-type mice were cocultured in transwell plates, the phagocytic index of macrophages was not altered demonstrating that B-1 cells can influence the effector functions of macrophages in vitro via IL-10 secretion.

EARLY ACTIVATION OF SPLENIC MACROPHAGES BY TUMOR-NECROSIS-FACTOR-ALPHA IS IMPORTANT IN DETERMINING THE OUTCOME OF EXPERIMENTAL HISTOPLASMOSIS IN MICE

Experimental infection of animals with Histoplasma capsulatum caused a massive macrophage infiltration into the spleen and induced the production of tumor necrosis factor alpha (TNF-alpha) locally. The cytokine was also produced in vitro by peritoneal exudate macrophages exposed to a large inoculum of yeast cells. Depletion of the cytokine by injection of polyclonal sheep anti-TNF-alpha antibody was detrimental to sublethally infected mice. Fungous burdens in the spleens of TNF-alpha-depleted mice were higher than they were in the infected control mice at days 2, 7, and 9 after infection, and the antibody-treated animals succumbed to the infection. Histopathological study of spleen sections revealed that splenic macrophages were not able to control proliferation of intracellular yeasts as a result of TNF-alpha depletion. It seems that TNF-alpha plays a role in early activation of splenic macrophages which is important in controlling the outcome of an infection.

RESPONSE TO TEMPERATURE IN THE OXYGEN-UPTAKE OF AWAKE AND DORMANT FROGS (AMPHIBIA, LEPTODACTYLIDAE)

In southeastern Brazil the leptodactylid frogs Leptodactylus fuscus and Physalaemus fuscomaculatus enter dormancy during the dry season. Oxygen uptake was measured in awake and dormant groups of both species in a temperature range at which these frogs are usually exposed throughout the year. Oxygen uptake was lower for dormant groups at high temperature, and a lack of response to temperature was reported between 20 - 25 C in the dormant group of both species. This temperature-intensitive range can be considered an adaptive feature to save fat reserves during dormancy.

EFFECT OF A PERSISTENT INFLAMMATORY PROCESS ON EXPERIMENTAL HETEROTOPIC OSSIFICATION - THE INFLUENCE OF MACROPHAGES

1. We investigated the effect of a persistent carrageenin- or nystatin-induced inflammatory reaction on heterotopic ossification produced by the subcutaneous implant of a demineralized bone matrix in female Swiss mice (25 to 35 g).2. Subcutaneous carrageenin injection (0.3 ml of a 2% solution in saline) into mice induced an inflammatory reaction characterized by a mature granuloma predominantly of macrophages containing particles of the irritant in their cytoplasm and which remained unchanged until the end of the experiment (40th day).3. Subcutaneous nystatin inoculation (30,000 IU in 0.3 ml saline) induced an inflammatory reaction consisting initially of macrophages (4th day) but later turning into an epithelioid granuloma (7th day) consisting predominantly of epithelioid cells and which was present up to the 2 lst day when it was gradually replaced by adipocytes up to the 30th day.4. An intramuscular implant of demineralized bone matrix (DBM, approximately 10 mg) induced the formation of cartilage and bone tissue and of hemopoietic bone marrow (heterotopic ossification) in 100% of the control animals (N = 5). An intramuscular DBM implant in animals that received carrageenin (N = 19) or nystatin (N = 21) induced heterotopic ossification in 100 and 57% (P<0.01)) of the animals, respectively.5. The response to a dorsal subcutaneous DBM implant was essentially negative in control animals (N = 5), whereas implants performed near the site injected with carrageenin (N = 28) or nystatin (N = 31) produced a response in 71 (P <0.01) and 36 % (P<0.01) of the animals, respectively. A DBM implant into the contralateral (control) dorsal subcutaneous tissue of the same animals that received carrageenin (N = 25) or nystatin (N = 29) resulted in heterotopic ossification in 64 (P<0.01) and 7% of the animals, respectively.6. The results suggest that the macrophages present in the mature granuloma induced by carrageenin somehow favored the development of metaplastic plates after subcutaneous DBM implant and that this effect may be systemic since the same response was observed in contralateral subcutaneous tissue.

SUPPORT OF PARACOCCIDIOIDES-BRASILIENSIS MULTIPLICATION BY HUMAN MONOCYTES OR MACROPHAGES - INHIBITION BY ACTIVATED PHAGOCYTES

The interaction of human monocytes or monocyte-derived macrophages and yeast-form Paracoccidioides brasiliensis was studied in vitro. Yeast cells were readily ingested by adherent monocytes or macrophages. Multiplication of P. brasiliensis, measured by growth as colony forming units (cfu) on a supplemented medium with good plating efficiency, was greater in monocyte co-cultures compared to the number of cfu obtained from complete tissue-culture medium (CTCM). Multiplication increased with time in macrophage cocultures, e.g., from two-six-fold in 24 h to nine-fold in 72 h. Microscopic observations indicated that ingested yeast cells multiplied inside macrophages. When monocytes were treated with supernate cytokines (CK) from concanavalin-A-stimulated mononuclear cells, then co-cultured with P. brasiliensis, multiplication was significantly inhibited compared with control monocyte co-cultures. Treatment of macrophages-derived from monocytes by culture in vitro for 3 days-for a further 3 days with CK resulted in maximal inhibition of multiplication over the subsequent 72 h. Similarly, when monocyte-derived macrophages (after culture for 7 days) were treated for 3 days with recombinant human gamma-interferon (IFN; 300 U/ml) or CK they restricted multiplication of P. brasiliensis by 65% and 95%, respectively, compared with control macrophages, Antibody to IFN abrogated the effect of IFN or CK treatment. These findings show that ingested P. brasiliensis can multiply in human monocytes or macrophages and that this multiplication can be restricted by activated monocytes or macrophages.

Effects of aerobic endurance training status and specificity on oxygen uptake kinetics during maximal exercise

The main purpose of this study was to analyze the effects of exercise mode, training status and specificity on the oxygen uptake ((V)over dot O-2) kinetics during maximal exercise performed in treadmill running and cycle ergometry. Seven runners (R), nine cyclists (C), nine triathletes (T) and eleven untrained subjects (U), performed the following tests on different days on a motorized treadmill and on a cycle ergometer: (1) incremental tests in order to determine the maximal oxygen uptake ((V)over dot O-2max) and the intensity associated with the achievement of (V)over dot O-2max (I(V)over dot O-2max); and (2) constant work-rate running and cycling exercises to exhaustion at I(V)over dot O-2max to determine the effective time constant of the (V)over dot O-2 response (tau(V)over dot O-2). Values for (V)over dotO(2max) obtained on the treadmill and cycle ergometer [R=68.8 (6.3) and 62.0 (5.0); C=60.5 (8.0) and 67.6 (7.6); T=64.5 (4.8) and 61.0 (4.1); U=43.5 (7.0) and 36.7 (5.6); respectively] were higher for the group with specific training in the modality. The U group showed the lowest values for VO2max, regardless of exercise mode. Differences in tau(V)over dot O-2 (seconds) were found only for the U group in relation to the trained groups [R=31.6 (10.5) and 40.9 (13.6); C=28.5 (5.8) and 32.7 (5.7); T=32.5 (5.6) and 40.7 (7.5); U=52.7 (8.5) and 62.2 (15.3); for the treadmill and cycle ergometer, respectively]; no effects of exercise mode were found in any of the groups. It is concluded that tauVO(2) during the exercise performed at I(V)over dot O-2max is dependent on the training status, but not dependent on the exercise mode and specificity of training. Moreover, the transfer of the training effects on tau(V)over dotO(2) between both exercise modes may be higher compared with (V)over dot O-2max.