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Aims: Balloon cryoablation is an accepted method of achieving pulmonary vein isolation for the treatment of atrial fibrillation. The relationship between blood flow in the atrium and cryo energy delivery to the tissue remains poorly understood., Methods and Results: Controlled cryoablations were performed in vitro using a pulmonary vein phantom constructed from bovine muscle, providing a 20 mm vein ostium. A temperature sensor was mounted within the 'vein wall' at a 1 mm tissue depth. Apparatus was constructed to assess the effect of incomplete pulmonary venous occlusion causing a leak, simulated atrial stasis, atrial circulation, and mitral regurgitation. Controlled ablations using the 2nd generation 28 mm cryoballoon catheter were repeated three times and mean values compared. Leak volume significantly affected both balloon temperatures and tissue temperatures. Simulated mitral regurgitation (MR) significantly impaired the effectiveness of cryo energy delivery resulting in significantly warmer balloon and tissue temperatures. With high leak volumes and moderate to severe MR there was a marked disparity between the cryoballoon temperature and the tissue temperature of approximately 60 degrees. Balloon warming times varied inversely with both leak volume and simulated MR flow volume., Conclusion: Incomplete venous occlusion and MR result in warmer balloon and tissue temperatures, and shorter balloon warming times, and are likely to significantly impair the effectiveness of cryoablation. Balloon temperature is poor indicator of tissue temperature under higher flow conditions.

Carbon dioxide (CO2) released from respiring cells in the stems of trees (RS) can diffuse radially to the atmosphere (EA) or dissolve in xylem sap and move internally in the tree (FT). Previous studies have observed that EA decreases as stem or branch diameter increases, but the cause of this relationship has not been determined, nor has the relationship been confirmed between stem diameter and RS, which includes both EA and FT. In this study, for the first time the mass balance technique was used to estimate RS of stems of Liriodendron tulipifera L. trees of different diameters, ranging from 16 to 60 cm, growing on the same site. The magnitude of the component fluxes scaled with tree size. Among the five trees, the contribution of EA to RS decreased linearly with increasing stem diameter and sapwood area while the contribution of FT to RS increased linearly with stem diameter and sapwood area. For the smallest tree EA was 86% of RS but it was only 46% of RS in the largest tree. As tree size increased a greater proportion of respired CO2 dissolved in sap and remained within the tree. Due to increase in FT with tree size, we observed that trees of different sizes had the same RS even though they had different EA. This appears to explain why the EA of stems and branches decreases as their size increases., (© The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Staphylococcus aureus iron-regulated surface protein A (IsdA) is a fibrinogen and fibronectin adhesin that also contributes to iron sequestration and resistance to innate immunity. IsdA is conserved in human isolates and has been investigated as a human vaccine candidate. Here we report the expression of isdA, the efficacy of anti-IsdA responses and the existence of IsdA sequence variants from bovine Staphylococcus. Clinical staphylococci were obtained from US dairy farms and assayed by PCR for the presence and expression of isdA. isdA-positive species from bovines included S. aureus, S. haemolyticus and S. chromogenes. Immunoassays on bovine milk and serum confirmed the induction and opsonophagocytic activity of anti-IsdA humoral responses. The variable region of isdA was sequenced and protein alignments predicted the presence of two main variants consistent with those from human S. aureus. Mouse antibodies against one IsdA variant reduced staphylococcal binding to fibronectin in vitro in an isotype-dependent manner. Purified IsdA variants bound distinctly to fibronectin and fibrinogen. Our findings demonstrate that variability within the C-terminus of this adhesin affects immune reactivity and binding specificity, but are consistent with the significance of IsdA in bovine disease and relevant for vaccine development., (© FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The effect of temperature on stomatal conductance (gs) and corresponding gas exchange parameters was studied in two tree species with contrasting leaf anatomy and ecophysiology-a broadleaf angiosperm, Populus deltoides x nigra (poplar), and a needle-leaf gymnosperm, Pinus taeda (loblolly pine). Experiments were conducted in growth chambers across a leaf temperature range of 19-48°C. Manipulations of temperature were done in well-watered and drought soil conditions and under ambient (400 ppm) and elevated (800 ppm) air CO2 concentrations. Increases in leaf temperature caused stomatal opening at both ambient and elevated [CO2]. The gs increased by 42% in poplar and by 40% in loblolly pine when leaf temperature increased from 30°C to 40°C at a vapour pressure difference of 1 kPa. Stomatal limitation to photosynthesis decreased in elevated temperature in loblolly pine but not in poplar. The ratio of net photosynthesis to gs depended on leaf temperature, especially at high temperatures. Evaporative cooling of transpiring leaves resulted in reductions in leaf temperature up to 9°C in well-watered poplar but only 1°C in drought-stressed poplar and in loblolly pine. As global mean temperatures rise and temperature extremes become more frequent and severe, understanding the effect of temperature on gs, and modelling that relationship, will become increasingly important., (© The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

To accurately estimate stem respiration (R S ), measurements of both carbon dioxide (CO 2 ) efflux to the atmosphere (E A ) and internal CO 2 flux through xylem (F T ) are needed because xylem sap transports respired CO 2 upward. However, reports of seasonal dynamics of F T and E A are scarce and no studies exist in Mediterranean species under drought stress conditions. Internal and external CO 2 fluxes at three stem heights, together with radial stem growth, temperature, sap flow and shoot water potential, were measured in Quercus pyrenaica Willd. in four measurement campaigns during one growing season. Substantial daytime depressions in temperature-normalized E A were observed throughout the experiment, including prior to budburst, indicating that diel hysteresis between stem temperature and E A cannot be uniquely ascribed to diversion of CO 2 in the transpiration stream. Low internal [CO 2 ] (<0.5%) resulted in low contributions of F T to R S throughout the growing season, and R S was mainly explained by E A (>90%). Internal [CO 2 ] was found to vary vertically along the stems. Seasonality in resistance to radial CO 2 diffusion was related to shoot water potential. The low internal [CO 2 ] and F T observed in our study may result from the downregulation of xylem respiration in response to a legacy of coppicing as well as high radial diffusion of CO 2 through cambium, phloem and bark tissues, which was related to low water content of stems. Long-term studies analyzing temporal and spatial variation in internal and external CO 2 fluxes and their interactions are needed to mechanistically understand and model respiration of woody tissues., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Since a substantial portion of respired CO2 remains within the stem, diel and seasonal trends in stem CO2 concentration ([CO2]) are of major interest in plant respiration and carbon budget research. However, continuous long-term stem [CO2] studies are scarce, and generally absent in Mediterranean climates. In this study, stem [CO2] was monitored every 15min together with stem and air temperature, sap flow, and soil water storage during a growing season in 16 stems of Quercus pyrenaica to elucidate the main drivers of stem [CO2] at different temporal scales. Fluctuations in sap pH were also assessed during two growing seasons to evaluate potential errors in estimates of the concentration of CO2 dissolved in xylem sap ([CO2*]) calculated using Henry's law. Stem temperature was the best predictor of stem [CO2] and explained more than 90% and 50% of the variability in stem [CO2] at diel and seasonal scales, respectively. Under dry conditions, soil water storage was the main driver of stem [CO2]. Likewise, the first rains after summer drought caused intense stem [CO2] pulses, suggesting enhanced stem and root respiration and increased resistance to radial CO2 diffusion. Sap flow played a secondary role in controlling stem [CO2] variations. We observed night-time sap pH acidification and progressive seasonal alkalinization. Thus, if the annual mean value of sap pH (measured at midday) was assumed to be constant, night-time sap [CO2*] was substantially overestimated (40%), and spring and autumn sap [CO2*] were misestimated by 25%. This work highlights that diel and seasonal variations in temperature, tree water availability, and sap pH substantially affect xylem [CO2] and sap [CO2*]., (© The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Aims: The phrenic nerves may be damaged during catheter ablation of atrial fibrillation. Phrenic nerve function is routinely monitored during ablation by stimulating the right phrenic nerve from a site in the superior vena cava (SVC) and manually assessing the strength of diaphragmatic contraction. However the optimal stimulation site, method of assessing diaphragmatic contraction, and techniques for monitoring the left phrenic nerve have not been established. We assessed novel techniques to monitor phrenic nerve function during cryoablation procedures., Methods and Results: Pacing threshold and stability of phrenic nerve capture were assessed when pacing from the SVC, left and right subclavian veins. Femoral venous pressure waveforms were used to monitor the strength of diaphragmatic contraction. Stable capture of the left phrenic nerve by stimulation in the left subclavian vein was achieved in 96 of 100 patients, with a median capture threshold of 2.5 mA [inter-quartile range (IQR) 1.4-5.0 mA]. Stimulation of the right phrenic nerve from the subclavian vein was superior to stimulation from the SVC with lower pacing thresholds (1.8 mA IQR 1.4-3.3 vs. 6.0 mA IQR 3.4-8.0, P < 0.001). Venous pressure waveforms were obtained in all patients and attenuation of the waveform was always observed prior to onset of phrenic nerve palsy., Conclusion: The left phrenic nerve can be stimulated from the left subclavian vein. The subclavian veins are the optimal sites for phrenic nerve stimulation. Monitoring the femoral venous pressure waveform is a novel technique for detecting impending phrenic nerve damage., (Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2014. For permissions please email: journals.permissions@oup.com.)

The effect of transpiration rate on internal assimilation of CO2 released from respiring cells has not previously been quantified. In this study, detached branches of Populus deltoides were allowed to take up (13)CO2-labelled solution at either high (high label, HL) or low (low label, LL) (13)CO2 concentrations. The uptake of the (13)CO2 label served as a proxy for the internal transport of respired CO2, whilst the transpiration rate was manipulated at the leaf level by altering the vapour pressure deficit (VPD) of the air. Simultaneously, leaf gas exchange was measured, allowing comparison of internal CO2 assimilation with that assimilated from the atmosphere. Subsequent (13)C analysis of branch and leaf tissues revealed that woody tissues assimilated more label under high VPD, corresponding to higher transpiration, than under low VPD. More (13)C was assimilated in leaf tissue than in woody tissue under the HL treatment, whereas more (13)C was assimilated in woody tissue than in leaf tissue under the LL treatment. The ratio of (13)CO2 assimilated from the internal source to CO2 assimilated from the atmosphere was highest for the branches under the HL and high VPD treatment, but was relatively small regardless of VPD×label treatment combination (up to 1.9%). These results showed that assimilation of internal CO2 is highly dependent on the rate of transpiration and xylem sap [CO2]. Therefore, it can be expected that the relative contribution of internal CO2 recycling to tree carbon gain is strongly dependent on factors controlling transpiration, respiration, and photosynthesis.

Predicted future changes in air temperature and atmospheric CO(2) concentration ([CO(2)]), coupled with altered precipitation, are expected to substantially affect tree growth. Effects on growth may vary considerably across a species range, as temperatures vary from sub-optimal to supra-optimal for growth. We performed an experiment simultaneously at two locations in the current range of loblolly pine, a cool site and a warm site, to examine the effect of future climate conditions on growth of loblolly pine seedlings in contrasting regions of the species range. At both sites 1-year-old loblolly pine seedlings were grown in current (local ambient temperature and [CO(2)]) and predicted future atmospheric conditions (ambient +2 °C temperature and 700 μmol mol(-1) [CO(2)]). Additionally, high and low soil moisture treatments were applied within each atmospheric treatment at each site by altering the amount of water provided to the seedlings. Averaged across water treatments, photosynthesis (A(net)) was 31% greater at the cool site and 34% greater at the warm site in elevated temperature and [CO(2)] compared with ambient temperature. Biomass accumulation was also stimulated by 38% at the cool site and by 24% at the warm site in that treatment. These results suggest that a temperature increase of 2 °C coupled with an increase in [CO(2)] (predicted future climate) will create conditions favorable for growth of this species. Reduced soil moisture decreased growth in both current and predicted atmospheric conditions. Biomass accumulation and A(net) were reduced by ∼39 and 17%, respectively, in the low water treatment. These results suggest that any benefit of future atmospheric conditions may be negated if soil moisture is reduced by altered precipitation patterns.

If an increase in temperature will limit the growth of a species, it will be in the warmest portion of the species distribution. Therefore, in this study we examined the effects of elevated temperature on net carbon assimilation and biomass production of northern red oak (Quercus rubra L.) seedlings grown near the southern limit of the species distribution. Seedlings were grown in chambers in elevated CO(2) (700 µmol mol(-1)) at three temperature conditions, ambient (tracking diurnal and seasonal variation in outdoor temperature), ambient +3 °C and ambient +6 °C, which produced mean growing season temperatures of 23, 26 and 29 °C, respectively. A group of seedlings was also grown in ambient [CO(2)] and ambient temperature as a check of the growth response to elevated [CO(2)]. Net photosynthesis and leaf respiration, photosynthetic capacity (V(cmax), J(max) and triose phosphate utilization (TPU)) and chlorophyll fluorescence, as well as seedling height, diameter and biomass, were measured during one growing season. Higher growth temperatures reduced net photosynthesis, increased respiration and reduced height, diameter and biomass production. Maximum net photosynthesis at saturating [CO(2)] and maximum rate of electron transport (J(max)) were lowest throughout the growing season in seedlings grown in the highest temperature regime. These parameters were also lower in June, but not in July or September, in seedlings grown at +3 °C above ambient, compared with those grown in ambient temperature, indicating no impairment of photosynthetic capacity with a moderate increase in air temperature. An unusual and potentially important observation was that foliar respiration did not acclimate to growth temperature, resulting in substantially higher leaf respiration at the higher growth temperatures. Lower net carbon assimilation was correlated with lower growth at higher temperatures. Total biomass at the end of the growing season decreased in direct proportion to the increase in growth temperature, declining by 6% per 1 °C increase in mean growing season temperature. Our observations suggest that increases in air temperature above current ambient conditions will be detrimental to Q. rubra seedlings growing near the southern limit of the species range.

Although many effects of staphylococcal superantigens (SAg) on T cells are well established, less is known about their effects on APC. In this study, bovine PBMC were stimulated with a low dose of staphylococcal enterotoxin C1 (SEC1). The phenotype of adherent cells (Ac) derived from bovine PBMC cultured with SEC1 [SEC1-stimulated Ac (sAc)] for 192 h was CD14(-), CD68(-), CD163(-), dendritic cell (DC)-specific ICAM-3-grabbing nonintegrin(+), MHC class II (MHC II)(high), CD11a(low), CD11b(high), CD11c(high), and CD1b(high), suggesting these cells were dendritic cells (DC). SEC1 also induced transcription of the CXCL1, -2, and -3 family, CXCL6, CCL2, and CCL5 genes in sAc, which increased rapidly but returned to basal levels by 48 h. In contrast, increased transcription of CCL3, CCL8, and CXCL12, responsible for mononuclear cell migration and chronic inflammation, was sustained. In vitro cell migration assays showed vigorous migration of granulocytes, followed by migration of mononuclear cells. The autologous MLR showed that sAc induced a dose-dependent proliferation of CD4(+) T cells and an even stronger proliferation of CD8(+) T cells. This effect was inhibited or reduced by pretreatment with mAb to CD11b, MHC II, or MHC II plus CD18. These results indicate that stimulation of bovine PBMC by SAg induces differentiation of monocytes into DC.

Previous reports have shown that CO(2) dissolved in xylem sap in tree stems can move upward in the transpiration stream. To determine the fate of this dissolved CO(2), the internal transport of respired CO(2) at high concentration from the bole of the tree was simulated by allowing detached young branches of sycamore (Platanus occidentalis L.) to transpire water enriched with a known quantity of (13)CO(2) in sunlight. Simultaneously, leaf net photosynthesis and CO(2) efflux from woody tissue were measured. Branch and leaf tissues were subsequently analysed for (13)C content to determine the quantity of transported (13)CO(2) label that was fixed. Treatment branches assimilated an average of 35% (SE=2.4) of the (13)CO(2) label taken up in the treatment water. The majority was fixed in the woody tissue of the branches, with smaller amounts fixed in the leaves and petioles. Overall, the fixation of internally transported (13)CO(2) label by woody tissues averaged 6% of the assimilation of CO(2) from the atmosphere by the leaves. Woody tissue assimilation rates calculated from measurements of (13)C differed from rates calculated from measurements of CO(2) efflux in the lower branch but not in the upper branch. The results of this study showed unequivocally that CO(2) transported in xylem sap can be fixed in photosynthetic cells in the leaves and branches of sycamore trees and provided evidence that recycling of xylem-transported CO(2) may be an important means by which trees reduce the carbon cost of respiration.

In spite of the importance of respiration in forest carbon budgets, the mechanisms by which physiological factors control stem respiration are unclear. An experiment was set up in a Eucalyptus globulus plantation in central Portugal with monoculture stands of 5-year-old and 10-year-old trees. CO(2) efflux from stems under shaded and unshaded conditions, as well as the concentration of CO(2) dissolved in sap [CO(2)(*)], stem temperature, and sap flow were measured with the objective of improving our understanding of the factors controlling CO(2) release from stems of E. globulus. CO(2) efflux was consistently higher in 5-year-old, compared with 10-year-old, stems, averaging 3.4 versus 1.3 mumol m(-2) s(-1), respectively. Temperature and [CO(2)(*)] both had important, and similar, influences on the rate of CO(2) efflux from the stems, but neither explained the difference in the magnitude of CO(2) efflux between trees of different age and size. No relationship was found between efflux and sap flow, and efflux was independent of tree volume, suggesting the presence of substantial barriers to the diffusion of CO(2) from the xylem to the atmosphere in this species. The rate of corticular photosynthesis was the same in trees of both ages and only reduced CO(2) efflux by 7%, probably due to the low irradiance at the stem surface below the canopy. The younger trees were growing at a much faster rate than the older trees. The difference between CO(2) efflux from the younger and older stems appears to have resulted from a difference in growth respiration rather than a difference in the rate of diffusion of xylem-transported CO(2).

Respiration of stems and branches of trees (R(S)) has typically been estimated by measuring radial CO(2) efflux from woody tissue (E(A)) and rates of efflux are often scaled temporally using a temperature relationship (Q(10)). High concentrations of CO(2) in xylem sap ([CO(2)*]) have been shown to affect E(A), and the transport of CO(2) in the xylem stream has been suggested as a mechanism to explain field observations of temperature-independent fluctuations in E(A). Sap velocity and temperature were manipulated in detached branch segments of sycamore (Platanus occidentalis L.) under controlled conditions to quantify these effects. Within individual branches of similar size, E(A) and [CO(2)*] were greater at low sap velocity, while the amount of respired CO(2) transported in sap (transport flux, F(T)) was greater at high sap velocity. E(A) was linearly correlated with [CO(2)*]. In branches of three diameter classes (1, 2, and 3 cm), volume-based E(A), F(T), and R(S) did not differ, but surface-area based CO(2) fluxes increased with diameter class. Regardless of diameter, E(A) accounted for only 30% of respired CO(2) at high sap velocity, while at low sap velocity, E(A) accounted for 71% of respired CO(2). E(A), F(T), and R(S) measured at 5, 20, and 35 degrees C at the same sap velocity showed a typical exponential response to temperature. However, at the lowest temperature, E(A) accounted for only 18% of the CO(2) released from respiring cells compared with 44% at the highest temperature, perhaps due to the effect of temperature on the solubility of CO(2) in water. These results directly demonstrate the transport of respired CO(2) in the xylem stream and may help to explain inconsistencies in stem and branch respiration measurements made in situ.

The respiration rate of a tree stem has commonly been estimated from measurements of CO2 efflux to the atmosphere. These estimates assume that all CO2 efflux originates from respiration of local tissues and that all CO2 produced by local tissues escapes to the atmosphere through the bark. However, dissolved CO2 can be transported in the xylem stream, and CO2 concentration ([CO2]) in xylem can be up to three orders of magnitude greater than that of the atmosphere, suggesting that measurements of CO2 efflux do not account for all CO2 produced by respiration. Here, we propose a new mass balance approach for estimating the respiration rate of tree stems that accounts for both external and internal fluxes of CO2. We demonstrate this approach using measurements of CO2 efflux, sap flux and internal [CO(2)] to calculate the rate of CO2 production of a segment of stem tissue in situ. At different times of the day, CO2 produced by respiration of stem tissues followed different flux pathways. During daylight hours when sap was flowing, a large proportion of respired CO2 was carried away in the xylem stream, whereas at night, most respiratory CO2 escaped to the atmosphere through the bark. Our calculations showed errors in efflux-based estimates of respiration of up to 76% compared with estimates that include both internal and external fluxes.

We monitored sap flux density (v) diurnally in nine mature southeastern pine (Pinus spp.) trees with a thermal dissipation probe that spanned the sapwood radius. We found the expected pattern of high v near the cambium and decreasing v with depth toward the center of the tree; however, the pattern was not constant within a day or between trees. Radial profiles of trees were steeper earlier in the day and became less steep later in the day. As a result, time-dependent changes in the shape of the radial profile of v were sometimes correlated with daily changes in evaporative demand. As the radial profile became less steep, the inner xylem contributed relatively more to total tree sap flow than it did earlier in the day. We present a 3-parameter Gaussian function that can be used to describe the radial distribution of v in trees. Parameters in the function represent depth in the xylem from the cambium, maximum v, depth in the xylem where maximum v occurs, and the rate of radial change in v with radial depth (beta). Values of beta varied significantly between trees and with time, and were sometimes correlated with air vapor pressure deficit (D). We hypothesize that this occurred during periods of high transpiration when the water potential gradient became great enough to move water in the inner sapwood despite its probable high hydraulic resistance. We examined discrepancies among estimates of daily water use based on single-point, two-point and multi-point (i.e., every 20 mm in the sapwood) measurements. When radial distribution of v was not considered, a single-point measurement resulted in errors as large as 154% in the estimate of daily water use relative to the estimate obtained from a multi-point measurement. Measuring v at two close sample points (10 and 30 mm) did not improve the estimate; however, estimates derived from v measured at two distant sample points (10 and 70 mm) significantly improved the estimate of daily water use, although errors were as great as 32% in individual trees. The variability in v with depth in the xylem, over time, and between trees indicates that measurements of the radial distribution of v are necessary to accurately estimate water flow in trees with large sapwood areas.

We developed a new microelectrode technique for measuring CO2 concentration ([CO2]) in xylem sap of trees. This technique enabled us to make rapid and continuous measurements of xylem sap [CO2] in situ. In this report, we discuss the methodology and establish the feasibility of the technique. We also describe calibration procedures, temperature sensitivity, field use and other characteristics of the microelectrodes. An example of data collected in the field is provided. Microelectrode calibration was accomplished at constant temperature in air of known [CO2]. When sampling temperature differed from calibration temperature, correction was necessary. We developed an equation to correct for temperatures between 15 and 35 degrees C when calibration was conducted at 25 degrees C. Equations based on Henry's Law were used to convert measured gas phase [CO2] (%) to concentration of all products of CO2 dissolved in sap (mmol l(-1)). We inserted microelectrodes into stems of three tree species to measure diurnal changes in [CO2] in the xylem sap. A diurnal pattern with depression during the day and elevation at night was observed. Mean daily [CO2] ranged from 1.6 to 10.3 mmol l(-1). Microelectrodes were suitable for making diurnal measurements for up to 7 days without recalibration. We also used the microelectrodes to measure [CO2] of soil in situ. Soil [CO2] ranged from 1 to 4% (gas phase), with little diurnal variation.

The insulin-like growth factors (IGF) circulate bound to specific high affinity binding proteins (IGFBPs) that modulate physiological responses to both IGF-I and IGF-II. Administration of bovine somatotropin (bST) to lactating cows has been shown to increase total serum levels of IGF-I; however, its effects on IGFBPs are unknown. Therefore, we determined the effects of bST on specific IGFBPs that are present in bovine serum and lymph. The results show that bovine serum contains IGFBPs with mol wt (Mr) estimates of 43,000, 39,000, 34,000, 29,000, and 24,000, as determined by ligand blotting. Using specific antisera, immunoblotting showed that the 43,000 and 39,000 Mr bands were IGFBP-3 and the 34,000 Mr band was IGFBP-2. All five forms of IGFBP were also present in afferent mammary lymph. To determine the effect of bST, four cows were treated with bST (40 mg/day) or vehicle for 12-day periods using a cross-over design. The intensity of the IGFBP-3 band increased 3.3 +/- 0.1-fold (mean +/- SE; P less than 0.01) by day 5 of bST treatment compared to that in controls. The intensity of the IGFBP-2 band decreased 3-fold. Serum levels of IGFBP-2, determined by RIA, decreased from 581 +/- 62 ng/ml during the control period to 156 +/- 52 ng/ml by day 5 of bST treatment (P less than 0.01). IGFBP-2 levels remained low for the entire 12-day treatment interval and rose to control levels within 4 days after cessation of bST. Results of a second study demonstrated that the decrease in IGFBP-2 concentrations in plasma observed during bST treatment (578 +/- 60 vs. 335 +/- 57 ng/ml) was paralleled by a decrease in IGFBP-2 levels in afferent mammary lymph (274 +/- 24 vs. 147 +/- 22 ng/ml). In contrast, the increase in IGFBP-3 levels observed in plasma during bST treatment by ligand blot analysis was not observed in lymph. In summary, bST increased serum levels of IGFBP-3 and decreased serum concentrations of IGFBP-2, while only IGFBP-2 levels were decreased in mammary lymph. Further studies are needed to determine whether these differences reflect differences in transport across capillaries or local production of specific forms of IGFBP.