Soil carbon stock and stability under Eucalyptus-based silvopasture and other land-use systems in the Cerrado biodiversity hotspot.

During the past few decades, commercial silvopastoral systems (SPS) with exotic Eucalyptus (hybrid) trees have become popular in the Brazilian Cerrado (savanna). With the increasing awareness about the role of carbon (C) storage in soils as a climate-change mitigation strategy and the relationship between the nature of soil aggregates and the soil's carbon sequestration potential, it is important to understand the influence of such SPS systems on soil organic carbon (SOC) storage. We studied C content in three aggregate size classes in six land-use systems on Oxisols in Minas Gerais, Brazil. The systems were planted forest, native secondary forest, managed pasture, and three 8-year-old SPS, differing in their tree-planting configurations. Eucalyptus hybrid was the tree in SPS and planted forest treatments, and Urochloa decumbens was the grass in SPS and pasture treatments. From each treatment, replicated soil samples were collected from four depth-classes (0-10, 10-30, 30-60, and 60-100 cm), fractionated by wet sieving into the three aggregate-size classes, 2000 to 250 µm, 250 to 53 µm, and <53 µm size classes representing macroaggregates, microaggregates, and silt + clay, respectively, and their C contents determined. Down to 1 m, total SOC stock values ranged from 260 Mg ha-1 under pasture to 167 Mg ha-1 under native forest, with 174 Mg ha-1 for Eucalyptus plantation and about 195 Mg ha-1 for the three SPS. Compared to the degraded native forest, the pasture system had significantly higher SOC in the whole soil and the aggregate size fractions, especially in the lower soil-depth classes. The lower SOC stock of Eucalyptus hybrid SPS compared to open pasture differs from the general trend of SPS having higher stock. Given that the Cerrado biome is a biodiversity hotspot, the use of native nitrogen-fixing trees, of which there are several, is worth investigating. In addition, the conversion from Eucalyptus monocultures to SPS could be considered as a strategy to increase the SOC stock.

Agroforestry systems and environmental quality: introduction.

Investments in agroforestry research during the past three decades-albeit modest-have yielded significant gains in understanding the role of trees on farmlands, and the ecological and economic advantages of integrated farming systems. While early research focused mostly on farm or local levels, broader-level ecosystem services of agroforestry systems (AFS) have raised high expectations in recent years. The nine papers included in this special collection deal with three of such environmental benefits of AFS: water-quality enhancement, carbon sequestration, and soil improvement. These benefits are based on the perceived ability of (i) vegetative buffer strips (VBS) to reduce surface transport of agrochemical pollutants, (ii) large volumes of aboveground and belowground biomass of trees to store high amounts of C deeper in the soil profile, and (iii) trees to enhance soil productivity through biological nitrogen fixation, efficient nutrient cycling, and deep capture of nutrients. The papers included have, in general, substantiated these premises and provided new insights. For example, the riparian VBS are reported to increase the reservoir life, in addition to reducing transport of agrochemicals; the variations in C storage in different soil-fraction sizes suggest that microaggregate (250-53 µm) dynamics in the soil could be a good indicator of its C-storage potential; and the use of vector analysis technique is recommended in AFS to avoid consequences of inaccurate and overuse of fertilizers. The papers also identified significant knowledge gaps in these areas. A common theme across all three environmental quality issues covered is that more and varied research datasets across a broad spectrum of conditions need to be generated and integrated with powerful statistical tools to ensure wide applicability of the results. Furthermore, appropriate management practices that are acceptable to the targeted land users and agroforestry practitioners need to be designed to exploit these environmental benefits. The relative newness of research in environmental quality of AFS will pose some additional challenges as well. These include the lack of allometric equations for tree-biomass determination, absence of standardized norms on soil sampling depth, and limitations of fixed-effect models arising from issues such as pseudo-replication and repeated measures that are common in studies on preexisting field plots. Overall, this special collection is a timely effort in highlighting the promise of AFS in addressing some of the environmental quality issues, and the challenges in realizing that potential.

Soil carbon storage in silvopasture and related land-use systems in the brazilian cerrado.

Silvopastoral management of fast-growing tree plantations is becoming popular in the Brazilian Cerrado (savanna). To understand the influence of such systems on soil carbon (C) storage, we studied C content in three aggregate size classes in six land-use systems (LUS) on Oxisols in Minas Gerais, Brazil. The systems were a native forest, a treeless pasture, 24- and 4-yr-old eucalyptus ( sp.) plantations, and 15- and 4-yr-old silvopastures of fodder grass plus animals under eucalyptus. From each system, replicated soil samples were collected from four depths (0-10, 10-20, 20-50, and 50-100 cm), fractionated into 2000- to 250-, 250- to 53-, and <53-µm size classes representing macroaggregates, microaggregates, and silt + clay, respectively, and their C contents determined. Macroaggregate was the predominant size fraction under all LUS, especially in the surface soil layers of tree-based systems. In general, C concentrations (g kg soil) in the different aggregate size fractions did not vary within the same depth. The soil organic carbon (SOC) stock (Mg C ha) to 1-m depth was highest under pasture compared with other LUS owing to its higher soil bulk density. The soils under all LUS had higher C stock compared with other reported values for managed tropical ecosystems: down to 1 m, total SOC stock values ranged from 461 Mg ha under pasture to 393 Mg ha under old eucalyptus. Considering the possibility for formation and retention of microaggregates within macroggregates in low management-intensive systems such as silvopasture, the macroaggregate dynamics in the soil seem to be a good indicator of its C storage potential.

Soil carbon storage in silvopastoral systems and a treeless pasture in northwestern Spain.

Soil particle size and land management practices are known to have considerable influence on carbon (C) storage in soils, but such information is lacking for silvopastoral systems in Spain. This study quantified the amounts of soil C stored at various depths to 100 cm under silvopastoral plots of radiata pine ( D. Don) and birch ( Roth) in comparison to treeless pasture in Galicia, Spain. Soils were fractionated into three size classes (<53, 53-250, and 250-2000 µm), and C stored in them and in the whole (nonfractionated) soil was determined. Overall, the C stock to 1 m ranged from 80.9 to 176.9 Mg ha in these soils. Up to 1 m depth, 78.82% of C was found in the 0- to 25-cm soil depth, with 12.9, 4.92, and 3.36% in the 25- to 50-, 50- to 75-, and 75- to 100-cm depths, respectively. Soils under birch at 0 to 25 cm stored more C in the 250- to 2000-µm size class as compared with those under radiata pine; at that depth, pasture had more C than pine silvopasture in the smaller soil fractions (<53 and 53-250 µm). In the 75- to 100-cm depth, there was significantly more storage of C in the 250- to 2000-µm fraction in both silvopastures as compared with the pasture. The higher storage of soil C in larger fraction size in lower soil depths of silvopasture suggests that planting of trees into traditional agricultural landscapes will promote longer-term storage of C in the soil.

Soil carbon storage as influenced by tree cover in the Dehesa cork oak silvopasture of central-western Spain.

The extent of carbon (C) stored in soils depends on a number of factors including soil characteristics, climatic and other environmental conditions, and management practices. Such information, however, is lacking for silvopastoral systems in Spain. This study quantified the amounts of soil C stored at various depths (0-25, 25-50, 50-75, and 75-100 cm) under a Dehesa cork oak (Quercus suber L.) silvopasture at varying distances (2, 5, and 15 m) to trees. Soil C in the whole soil and three soil fractions (<53, 53-250, and 250-2000 µm) was determined. Results showed soil depth to be a significant factor in soil C stocks in all soil particle sizes. Distance to tree was a significant factor determining soil C stocks in the whole soil and the 250-2000 µm soil fraction. To 1 m depth, mean total C storage at 2, 5, and 15 m from cork oak was 50.2, 37, and 26.5 Mg ha(-1), respectively. Taking into account proportions of land surface area containing these C stocks at varying distances to trees to 1 m depth, with a tree density of 35 stems ha(-1), estimated landscape soil C is 29.9 Mg ha(-1). Greater soil C stocks directly underneath the tree canopy suggest that maintaining or increasing tree cover, where lost from disease or management, may increase long term storage of soil C in Mediterranean silvopastoral systems. The results also demonstrate the use of soil aggregate characteristics as better indicators of soil C sequestration potential and thus a tool for environmental monitoring.

Optical and thermal properties of edible coatings for application in solar drying.

Solar drying is a sustainable process that may impact the quality of dried food. This is because, pigments contained in food are sensitive to sunlight, and exposure to ultraviolet radiation can affect them. We applied biopolymer-based coatings on strawberry, from hydro-colloidal solutions of Opuntia ficus indica-mucilage, fenugreek, xanthan gum, gum Arabic, and guar gum to evaluate their potential use as UV filters for solar drying of food. Thermal properties and the optical transmittance, absorbance and reflectance of the coatings were measured to assess their influence on food-sunlight interaction. During the drying experiments, the moisture content, total anthocyanins (TA), and total phenolic compounds (TPC) were measured. Optical and thermal properties are influenced by the biopolymer-based coatings. Also, the optical properties are influenced by the coating thickness. The differences in optical and thermal properties influence the drying process. Differences exist in the drying rate for strawberry slices with coating, compared with those without the coatings. In general, the TA and TPC content in the product are better preserved under solar drying than in control experiments done in a drying oven. A partial transmittance of solar UV radiation is recommended to obtain increased TA and TPC contents in the dried product.

Carbon storage in soil size fractions under two cacao agroforestry systems in Bahia, Brazil.

Shaded perennial agroforestry systems contain relatively high quantities of soil carbon (C) resulting from continuous deposition of plant residues; however, the extent to which the C is sequestered in soil will depend on the extent of physical protection of soil organic C (SOC). The main objective of this study was to characterize SOC storage in relation to soil fraction-size classes in cacao (Theobroma cacao L.) agroforestry systems (AFSs). Two shaded cacao systems and an adjacent natural forest in reddish-yellow Oxisols in Bahia, Brazil were selected. Soil samples were collected from four depth classes to 1 m depth and separated by wet-sieving into three fraction-size classes (>250 microm, 250-53 microm, and <53 microm)-corresponding to macroaggregate, microaggregate, and silt-and-clay size fractions-and analyzed for C content. The total SOC stock did not vary among systems (mean: 302 Mg/ha). On average, 72% of SOC was in macroaggregate-size, 20% in microaggregate-size, and 8% in silt-and-clay size fractions in soil. Sonication of aggregates showed that occlusion of C in soil aggregates could be a major mechanism of C protection in these soils. Considering the low level of soil disturbances in cacao AFSs, the C contained in the macroaggregate fraction might become stabilized in the soil. The study shows the role of cacao AFSs in mitigating greenhouse gas (GHG) emission through accumulation and retention of high amounts of organic C in the soils and suggests the potential benefit of this environmental service to the nearly 6 million cacao farmers worldwide.

Anticancer effects of Annona glabra plant extracts in human leukemia cell lines.

Annona glabra (pond apple), a tropical tree growing wild in the Americas and Asia, is used in traditional medicine against several human ailments, including cancer. To validate the ethnopharmacological claims against cancer, the anticancer effects of alcoholic extracts prepared from pond apple leaves, pulp and seed, were investigated in human leukemia cell lines. The alcoholic extracts were not cytotoxic to normal human lymphocytes. However, extracts were highly cytotoxic to drug sensitive (CEM) and multidrug-resistant leukemia (CEM/VLB) cell lines. The seed extract was more potent than leaf and pulp extracts, and the cytotoxicity values were significantly lower than that for adriamycin. The seed extract caused a concentration-dependent increase in the percentage of the sub G0/G1, as well as G0/G1 cell population, contributing to the cytotoxicity. The sub G0/G1 population increased from 2.2 to 7.0% in CEM and from 1.0 to 10.7% in CEM/VLB cell lines, when the cells were treated with 0-10 Bg/ml seed extract. Treatment of CEM and CEM/VLB cells with seed extract induced apoptosis and necrosis in both sensitive and resistant leukemia cells in a concentration-dependent manner. The seed extract at 2 and 5 Bg/ml enhanced cellular daunorubicin accumulation, indicating the competitive P-glycoprotein binding ability and drug-resistance reversal effect. Treatment of CEM and CEM/VLB cells with seed extracts also up-regulated the expression of cyclin kinase inhibitor (WAF1/p21) contributing to the arrest of cells at the G0/G1 phase of the cell cycle. These results support the traditional use of A. glabra and the alcoholic seed extract is a potent source of anticancer compounds that could be utilized pharmaceutically.

Carbon storage of different soil-size fractions in Florida silvopastoral systems.

Compared with open (treeless) pasture systems, silvopastoral agroforestry systems that integrate trees into pasture production systems are likely to enhance soil carbon (C) sequestration in deeper soil layers. To test this hypothesis, total soil C contents at six soil depths (0-5, 5-15, 15-30, 30-50, 50-75, and 75-125 cm) were determined in silvopastoral systems with slash pine (Pinus elliottii) + bahiagrass (Paspalum notatum) and an adjacent open pasture (OP) with bahiagrass at four sites, representing Spodosols and Ultisols, in Florida. Soil samples from each layer were fractionated into three classes (250-2000, 53-250, and <53 microm), and the C contents in each were determined. Averaged across four sites and all depths, the total soil organic carbon (SOC) content was higher by 33% in silvopastures near trees (SP-T) and by 28% in the alleys between tree rows (SP-A) than in adjacent open pastures. It was higher by 39% in SP-A and 20% in SP-T than in open pastures in the largest fraction size (250-2000 microm) and by 12.3 and 18.8%, respectively, in the intermediate size fraction (53-250 microm). The highest SOC increase (up to 45 kg m(-2)) in whole soil of silvopasture compared with OP was at the 75- to 125-cm depth at the Spodosol sites. The results support the hypothesis that, compared with open pastures, silvopastures contain more C in deeper soil layers under similar ecological settings, possibly as a consequence of a major input to soil organic matter from decomposition of dead tree-roots.

Investigation of cytokine expression in human leukocyte cultures with two immune-modulatory homeopathic preparations.

BACKGROUND: The efficacy of homeopathic medicines for maintaining human health and treating disease has been extensively examined in clinical trials. However, there is a paucity of preclinical evaluations of the effects of homeopathic medicinal preparations on cellular signaling pathways relevant to the applications of these preparations. MATERIALS AND METHODS: In this study, the immune-modulatory effects of Phase 6 (for the stimulation of the nonspecific defense system) and Flu Terminator (for influenza and viral diseases) (Be Well Homeopathics Inc. Miami, FL), two homeopathic preparations developed for the purpose, were evaluated in normal human leukocyte cultures in vitro. RESULTS: Both Phase 6 and Flu Terminator stimulated the production of pro-and anti-inflammatory cytokines by human leukocytes, although higher doses often produced a weaker response than lower doses. The carrier solvent (20% ethanol) failed to elicit any cytokine synthesis. CONCLUSIONS: The results of the in vitro studies suggested that ultralow concentrations of ingredients in Phase 6 and Flu Terminator were capable of eliciting a human immune response.

Biochar in the Agroecosystem-Climate-Change-Sustainability Nexus.

Interest in the use of biochar in agriculture has increased exponentially during the past decade. Biochar, when applied to soils is reported to enhance soil carbon sequestration and provide other soil productivity benefits such as reduction of bulk density, enhancement of water-holding capacity and nutrient retention, stabilization of soil organic matter, improvement of microbial activities, and heavy-metal sequestration. Furthermore, biochar application could enhance phosphorus availability in highly weathered tropical soils. Converting the locally available feedstocks and farm wastes to biochar could be important under smallholder farming systems as well, and biochar use may have applications in tree nursery production and specialty-crop management. Thus, biochar can contribute substantially to sustainable agriculture. While these benefits and opportunities look attractive, several problems, and bottlenecks remain to be addressed before widespread production and use of biochar becomes popular. The current state of knowledge is based largely on limited small-scale studies under laboratory and greenhouse conditions. Properties of biochar vary with both the feedstock from which it is produced and the method of production. The availability of feedstock as well as the economic merits, energy needs, and environmental risks-if any-of its large-scale production and use remain to be investigated. Nevertheless, available indications suggest that biochar could play a significant role in facing the challenges posed by climate change and threats to agroecosystem sustainability.

Mechanism of macrophage activation by (1,4)-alpha-D-glucan isolated from Tinospora cordifolia.

The signaling mechanism of the novel (1,4)-alpha-D-glucan (RR1) isolated from the medicinal plant Tinospora cordifolia was investigated in macrophages to evaluate its immunostimulating properties. When RAW264.7 macrophages were incubated with RR1 at 4 degrees C, the novel glucan inhibited the phagocytosis of unopsonized zymosan A bioparticles in a dose-dependent manner. RR1 also inhibited the binding and internalization of opsonized zymosan A bioparticles, although at a lower level than laminarin. Incubation of macrophages with anti-CD11b mAb followed by RR1 failed to show any inhibitory effect on RR1-induced TNF-alpha synthesis confirming that complement receptor 3 (CR3) is not involved in the opsonic binding and internalization of RR1 in macrophages unlike zymosan A. The anti-CD11b mAb has significant inhibitory effect on the zymosan A-induced tumor necrosis factor (TNF)-alpha synthesis. RR1 induced TNF-alpha synthesis in macrophages in a dose-dependent manner which can be completely inhibited by the NF-kappaB inhibitor caffeic acid phenethyl ester (CAPE) or curcumin. RR1 activated NF-kappaB in a time- and dose-dependent manner and this modulation of nuclear NF-kappaB activity is associated with the degradation of I-kappaB alpha thus facilitating the translocation of NF-kappaB into the nucleus. RR1-induced NF-kappaB activity peaks at 8 h of RR1 stimulation while I-kappaB alpha degradation occurred within 1 h of stimulation. RR1-induced NF-kappaB activation occurred through TLR6 signaling as evidenced by the synthesis of IL-8 in TLR6-transfected HEK293 cells. These results show that the novel (1,4)-alpha-D-glucan from Tinospora cordifolia activates the immune system through the activation of macrophages that occurs through TLR6 signaling, NF-kappaB translocation and cytokine production.

Improved tuberculosis case detection through public-private partnership and laboratory-based surveillance, Kannur District, Kerala, India, 2001-2002.

BACKGROUND: Efforts to intensify global tuberculosis (TB) control are limited by difficulties in coordinating with private doctors. More than half of Indian TB patients may initially consult a private provider, but many are neither diagnosed accurately nor treated effectively. We established and evaluated a public-private partnership based on surveillance of TB detected in private laboratories and use of standardised directly observed treatment regimens. METHODS: In one district, the governmental TB control programme offered training in microscopy to all large private sector laboratories, and educated private physicians on the importance of microscopy for TB diagnosis. We reviewed records from participating private laboratories and all publicly diagnosed patients. RESULTS: Of 2328 pulmonary TB patients registered from July 2001 to December 2002, 404 (17%) were detected in the private sector. The annual new AFB-positive case notification rate increased by 21%, from 27.8/100,000 in 2000 to 33.5/100,000 in 2002. Surveillance at private laboratories found an additional 260 nonregistered AFB-positive patients. CONCLUSIONS: This public-private partnership substantially increased TB case detection and established a sustainable framework for private sector involvement in TB control. In the setting of a strong public sector programme, the combination of active surveillance of private laboratories along with physician sensitisation is a promising approach to improve TB case detection.

Expression profiles of apoptotic genes induced by curcumin in human breast cancer and mammary epithelial cell lines.

Curcumin (diferuloyl methane), the yellow-colored dietary pigment from the rhizomes of turmeric, has been recognized as a chemopreventive agent because of its antitumor, antioxidant and antiproliferative effects. The cytotoxic, apoptotic and gene regulatory effects of both turmeric and curcumin were investigated in the MCF-7 human breast cancer carcinoma cell line and compared with the effects in MCF-10A human mammary epithelial cells. MCF-7 cells were more sensitive to turmeric and curcumin than MCF-10A cells. MCF-10A cells retained comparatively less curcumin in the medium than MCF- 7 cells after 24 h, thereby reducing the cytotoxic effect. Curcumin induced a significantly higher percentage of apoptosis in MCF-7 than MCF-10A cells at all doses. Microarray hybridization of Clonetech apoptotic arrays with labeled first-strand probes of total RNA was performed to identify and characterize the genes regulated by curcumin in tumor cells. Of the 214 apoptosis-associated genes in the array, the expression of 104 genes was altered by curcumin treatment. The gene expression was altered up to 14-fold levels in MCF-7 as compared to only up to 1.5-fold in the MCF-10A cell line by curcumin. Curcumin up-regulated (>3 fold) 22 genes and down-regulated (<3-fold) 17 genes at both 25 microg/ml and 50 microg/ml doses in the MCF-7 cell line. The up-regulated genes include HIAP1, CRAF1, TRAF6, CASP1, CASP2, CASP3, CASP4, HPRT, GADD45, MCL-1, NIP1, BCL2L2, TRAP3, GSTP1, DAXX, PIG11, UBC, PIG3, PCNA, CDC10, JNK1 and RBP2. The down-regulated genes were TRAIL, TNFR, AP13, IGFBP3, SARP3, PKB, IGFBP, CASP7, CASP9, TNFSF6, TRICK2A, CAS, TRAIL-R2, RATS1, hTRIP, TNFb and TNFRSF5. While a dose-dependent gene expression change was noticed in some genes, opposite regulatory effects were induced by different curcumin doses in three apoptotic genes. These results suggest that curcumin induces apoptosis in breast cancer cells by regulation of multiple signaling pathways, indicating its potential use for prevention and treatment of cancer.

Evidence for second metabolic pathway for O2 from PtiO2 measurements in denervated cat carotid body.

O2 microelectrode studies were conducted in the cat carotid body (CB) to investigate the hypothesis that there is a second, low affinity metabolic pathway for O2 in addition to classical oxidative metabolism. Tissue PO2 (PtiO2) and O2 disappearance rates (dPO2/dt) after brief blood flow occlusion were measured with recessed cathode microelectrodes (tip diameter less than 5 microns) at 150 sites in 15 normal cats (controls) and at 154 sites in 5 cats in which one CB had been denervated 2 or 3 days before the experiments. Mean PtiO2 was slightly higher in denervated CBs: 79.6 +/- 1.6 (SE) Torr compared with 76.4 +/- 2.0 Torr for controls (P = not significant). Mean dPO2/dt was 8.4% faster: -8.42 +/- 0.28 Torr/s compared with -7.77 +/- 0.43 Torr/s for controls (P less than 0.05). The O2 consumption rate (VO2), calculated from dPO2/dt correcting for cat oxyhemoglobin, was 7.5% higher: 1.62 and 1.51 ml.100 g-1.min-1, respectively, for denervated and control CBs (P less than 0.05). The apparent Michaelis-Menten constant, Kmapp (defined as the PtiO2 where dPO2/dt decreased by 50% from the initial rate during the first 3 s after occlusion) was determined for each O2 disappearance curve. After denervation, Kmapp decreased significantly by -47%: 12.0 +/- 1.3 Torr compared with 22.6 +/- 2.5 Torr for controls (P less than 0.01). The data provide evidence for a second metabolic pathway for O2 in the CB that loses its influence on VO2 after denervation.

Two-cytochrome metabolic model for carotid body PtiO2 and chemosensitivity changes after hemorrhage.

O2 microelectrode measurements were made in the cat carotid body (CB) at normal control blood pressures (C) and after hemorrhage (H) to reduce mean arterial blood pressure [C, 98.7 +/- 4.6 (SE) mmHg; H, 58.1 +/- 1.8; P less than 0.001; paired t test; n = 9 cats]. Mean tissue PO2 (PtiO2) was significantly lower (C, 78.4 +/- 3.0 Torr; H, 65.3 +/- 4.8; P less than 0.01). Except for two experiments with good autoregulation, the decrease in PtiO2 correlated with the reduction in blood pressure (r = 0.791, P less than 0.005). Measurements of O2 disappearance curves (DCs) and sinus nerve discharge (ND) were obtained after blood supply was occluded for 30-45 s (56 C DCs, 44 H DCs). Disappearance rates (dPO2/dt) were significantly slower after hemorrhage (C, -7.52 +/- 0.47 Torr/s; H, -6.60 +/- 0.44; P less than 0.01), decreasing by 0.656 Torr/s for each 10 Torr fall in PtiO2 (r = 0.626, P less than 0.05). Resting ND before occlusion increased during hypotension (11.6 +/- 2.9% of control, P less than 0.01) and correlated with the decrease in PtiO2 (r = -0.792, P less than 0.005). A computer simulation was performed for a two-cytochrome metabolic model with a second, low-O2-affinity oxidase in addition to normal oxidative metabolism. The effects of cat oxyhemoglobin and blood pH on the O2 DC measurement were also taken into account. The simulation for the two-cytochrome model was consistent with our experimental data and predicts reductions in blood flow and O2 metabolism with hypotension after hemorrhage that have similarities, as well as aspects that disagree, with previous reports in the literature.

An in vitro capillary system for studies on microcirculatory O2 transport.

An in vitro artificial capillary system has been developed for use in examining the O2 transport properties of free hemoglobin and erythrocytes. The artificial capillary was constructed by casting a thin film of transparent silicone rubber around a strand of tungsten wire that was 24 micron in diameter. After the rubber had polymerized, the wire was removed. Typical dimensions of the silicone rubber film were 170 micron thick, 1 cm wide, 5 mm long in the direction of flow, and a 27-micron lumen diameter. The artificial capillary bed was mounted on a microscope and perfused by either hemoglobin solutions or cell suspensions. Fractional saturation was measured as a function of axial position by a dual-wave-length microspectrophotometer, and the flow rate was regulated precisely by a syringe pump. O2 release experiments were carried out by suffusing the gas space surrounding the artificial capillary film with 100% N2 and perfusing with an oxygenated sample. O2 uptake experiments were carried out by suffusing the gas space with O2-N2 mixtures and perfusing with deoxygenated samples. The axial velocities were varied from 3 to 15 mm/s. The residence time (the time a particular red cell or hemoglobin molecule has spent in the capillary) for 50% oxygenation of a 4 mM (heme) deoxyhemoglobin solution was approximately 0.05 s at 37 degrees C when the gas space surrounding the capillary contained air. The corresponding time for 50% oxygenation of an equivalent red cell suspension was approximately 0.25 s.(ABSTRACT TRUNCATED AT 250 WORDS)

Physiological factors affecting O2 transport by hemoglobin in an in vitro capillary system.

O2 transport was examined by measuring the fractional saturation of concentrated hemoglobin solutions flowing through an artificial capillary that was approximately 27 micron in diameter and embedded in a silicone rubber film approximately 170 micron thick. The effects of pH, hemoglobin concentration, O2 tension, temperature, and organic phosphate were measured and analyzed quantitatively by a rigorous mathematical model that included the geometry of the capillary in the silicone film, parabolic flow velocity distributions inside the lumen, and cooperative O2 binding by hemoglobin. The rates of both oxygenation and deoxygenation were limited by diffusion and governed by the magnitude of the O2 gradient between the intracapillary fluid phase and the external gas space. In uptake experiments, O2 flux is determined primarily by the external O2 tension (16-160 mmHg in our experiments) because the internal O2 pressure is kept small due to chemical combination with hemoglobin. In release experiments, the external O2 tension is maintained at zero, and the transport rate is determined by the intracapillary partial pressure of O2 that is proportional to the O2 half-saturation pressure of hemoglobin value of the hemoglobin sample. As a result, factors that change the affinity of hemoglobin for O2, such as pH, temperature, and organic phosphate concentration, influence strongly the rate of O2 release but have little effect on the rate of O2 uptake. These properties are physiologically advantageous, since a decrease in pH or an increase in temperature during exercise increases both the rate and extent of deoxygenation while not altering the kinetics of oxygenation.

Immune stimulating properties of a novel polysaccharide from the medicinal plant Tinospora cordifolia.

An alpha-D-glucan (RR1) composed of (1-->4) linked back bone and (1-->6) linked branches with a molecular mass of >550 kDa and exhibiting unique immune stimulating properties is isolated and characterized from the medicinal plant Tinospora cordifolia. This novel polysaccharide is noncytotoxic and nonproliferating to normal lymphocytes as well as tumor cell lines at 0-1000 microg/ml. It activated different subsets of the lymphocytes such as natural killer (NK) cells (331%), T cells (102%), and B cells (39%) at 100 microg/ml concentration. The significant activation of NK cells is associated with the dose-dependent killing of tumor cells by activated normal lymphocytes in a functional assay. Immune activation by RR1 in normal lymphocytes elicited the synthesis of interleukin (IL)-1beta (1080 pg/ml), IL-6 (21,833 pg/ml), IL-12 p70 (50.19 pg/ml), IL-12 p40 (918.23 pg/ml), IL-18 (27.47 pg/ml), IFN- gamma (90.16 pg/ml), tumor necrosis factor (TNF)-alpha (2225 pg/ml) and monocyte chemoattractant protein (MCP)-1 (2307 pg/ml) at 100 microg/ml concentration, while it did not induce the production of IL-2, IL-4, IL-10, interferon (IFN)-alpha and TNF-beta. The cytokine profile clearly demonstrates the Th1 pathway of T helper cell differentiation essential for cell mediated immunity, with a self-regulatory mechanism for the control of its overproduction. RR1 also activated the complements in the alternate pathway, demonstrated by a stepwise increase in C3a des Arg components. Incidentally, RR1 stimulation did not produce any oxidative stress or inducible nitric oxide synthase (iNOS) in the lymphocytes or any significant increase in nitric oxide production. The water solubility, high molecular mass, activation of lymphocytes especially NK cells, complement activation, Th1 pathway-associated cytokine profile, together with a low level of nitric oxide synthesis and absence of oxidative stress confer important immunoprotective potential to this novel alpha-D-glucan.

Interpretation of oxygen disappearance curves measured in blood perfused tissues.

We have developed a two compartment (tissue and blood) lumped parameter model to interpret oxygen disappearance curves (O2 DCs) measured in vivo with PO2 microelectrodes in tissues which are perfused with blood. To include the properties of the oxyhemoglobin equilibrium curve (HEC), we used an algorithm we have recently developed for both standard and nonstandard conditions. The new blood and tissue model is more useful than a previous analysis using the Hill equation for blood and constant oxygen consumption (VO2). The model can be adapted for constant (zero-order) consumption, Michelis-Menten kinetics, or for double cytochrome systems. Examples for the former include brain, and for the latter, carotid body. The models are discussed in relationship to experimental microelectrode measurements in gerbil brain and in cat carotid body after blood flow occlusion.