Systematic update to the mammalian relative potency estimate database and development of best estimate toxic equivalency factors for dioxin-like compounds.

The World Health Organization (WHO) assesses potential health risks of dioxin-like compounds using Toxic Equivalency Factors (TEFs). This study systematically updated the relative potency (REP) database underlying the 2005 WHO TEFs and applied advanced methods for quantitative integration of study quality and dose-response. Data obtained from fifty-one publications more than doubled the size of the previous REP database (∼1300 datasets). REP quality and relevance for these data was assessed via application of a consensus-based weighting framework. Using Bayesian dose-response modeling, available data were modeled to produce standardized dose/concentration-response Hill curves. Study quality and REP data were synthesized via Bayesian meta-analysis to integrate dose/concentration-response data, author-calculated REPs and benchmark ratios. The output is a prediction of the most likely relationship between each congener and its reference as model-predicted TEF uncertainty distributions, or the 'best estimate TEF' (BE-TEF). The resulting weighted BE-TEFs were similar to the 2005 TEFs, though provide more information to inform selection of TEF values as well as to provide risk assessors and managers with information needed to quantitatively characterize uncertainty around TEF values. Collectively, these efforts produce an updated REP database and an objective, reproducible approach to support development of TEF values based on all available data.

Short communication: Development and evaluation of equations to predict growth of Holstein dairy heifers in a tropical climate.

The objective of this study was to develop and validate equations for estimating growth of dairy heifers using measures of withers height, body weight (BW), and age. Measures of BW and withers height of 207 Holstein heifers raised in a tropical climate were taken from birth to calving, totaling to 2,047 observations. To be included in the database, the heifer had to have at least 4 measures recorded. After that, 4 models were built and evaluated as follows: (1) a linear model of BW as function of age (BW~A), (2) a linear model of the BW-to-height ratio as function of age (BW:H~A), (3) a quadratic model, adjusted for a defined plateau, to describe height as function of age (H~A), and (4) an exponential growth model of BW as function of height (BW~H). A cross-validation procedure was performed to evaluate accuracy and precision of the models. The linear relationship of BW~A and BW:H~A were estimated, respectively, by the equations: BW = 42.65 + 0.62 × A and BW:H = 0.70 + 0.0041 × A, where BW is in kilograms, BW:H = BW-to-height ratio (kg/cm), and A = age (d). Using the quadratic plateau for the model H~A, a critical "x" value of ~806 d and a height plateau of 138.6 cm were identified. Therefore, the following equations for estimating the height of animals younger and older than 806 d, respectively, were developed: H = 78.15 + 0.150 × A - 0.00009 × A2 and H = 78.15 + 0.150 × cvx - 0.00009 × cvx2, where H = height (cm) and cvx = 806 (critical "x" value; given in days). Additionally, the exponential model of W~H was estimated by the following equation: BW = 4.25 × exp(0.034 ×H), where BW is in kilograms and H = height (cm). A cross validation demonstrated that all equations had very high accuracy and precision. Overall, these models demonstrated that BW and BW-to-height ratio increase linearly as a function of age, while BW follows an exponential growth pattern as a function of height. Additionally, the H~A model predicted that heifers achieve a maximum height of 138.6 cm at 806 d of age.

Primary afferent-driven presynaptic inhibition of C-fiber inputs to spinal lamina I neurons.

Presynaptic inhibition of primary afferent terminals is a powerful mechanism for controlling sensory information flow into the spinal cord. Lamina I is the major spinal nociceptive projecting area and monosynaptic input from C-fibers to this region represents a direct pathway for transmitting pain signals to supraspinal centers. Here we used an isolated spinal cord preparation to show that this pathway is under control of the afferent-driven GABAergic presynaptic inhibition. Presynaptic inhibition of C-fiber input to lamina I projection and local-circuit neurons is mediated by recruitment of Aß-, Aδ- and C-afferents. C-fiber-driven inhibition of C-fibers functions as a feedforward mechanism, by which the homotypic afferents control sensory information flow into the spinal cord and regulate degree of the primary nociceptive afferent activation needed to excite the second order neurons. The presynaptic inhibition of C-fiber input to lamina I neurons may be mediated by both synaptic and non-synaptic mechanisms, and its occurrence and extent are quite heterogeneous. This heterogeneity is likely to be reflective of involvement of lamina I neurons in diverse circuitries processing specific modalities of sensory information in the superficial dorsal horn. Thus, our results implicate both low- and high-threshold afferents in the modulation of C-fiber input into the spinal cord.

The gathering firestorm in southern Amazonia.

Wildfires, exacerbated by extreme weather events and land use, threaten to change the Amazon from a net carbon sink to a net carbon source. Here, we develop and apply a coupled ecosystem-fire model to quantify how greenhouse gas-driven drying and warming would affect wildfires and associated CO2 emissions in the southern Brazilian Amazon. Regional climate projections suggest that Amazon fire regimes will intensify under both low- and high-emission scenarios. Our results indicate that projected climatic changes will double the area burned by wildfires, affecting up to 16% of the region's forests by 2050. Although these fires could emit as much as 17.0 Pg of CO2 equivalent to the atmosphere, avoiding new deforestation could cut total net fire emissions in half and help prevent fires from escaping into protected areas and indigenous lands. Aggressive efforts to eliminate ignition sources and suppress wildfires will be critical to conserve southern Amazon forests.

Predicting enteric methane emission of dairy cows with milk Fourier-transform infrared spectra and gas chromatography-based milk fatty acid profiles.

The objective of the present study was to compare the prediction potential of milk Fourier-transform infrared spectroscopy (FTIR) for CH4 emissions of dairy cows with that of gas chromatography (GC)-based milk fatty acids (MFA). Data from 9 experiments with lactating Holstein-Friesian cows, with a total of 30 dietary treatments and 218 observations, were used. Methane emissions were measured for 3 consecutive days in climate respiration chambers and expressed as production (g/d), yield (g/kg of dry matter intake; DMI), and intensity (g/kg of fat- and protein-corrected milk; FPCM). Dry matter intake was 16.3 ± 2.18 kg/d (mean ± standard deviation), FPCM yield was 25.9 ± 5.06 kg/d, CH4 production was 366 ± 53.9 g/d, CH4 yield was 22.5 ± 2.10 g/kg of DMI, and CH4 intensity was 14.4 ± 2.58 g/kg of FPCM. Milk was sampled during the same days and analyzed by GC and by FTIR. Multivariate GC-determined MFA-based and FTIR-based CH4 prediction models were developed, and subsequently, the final CH4 prediction models were evaluated with root mean squared error of prediction and concordance correlation coefficient analysis. Further, we performed a random 10-fold cross validation to calculate the performance parameters of the models (e.g., the coefficient of determination of cross validation). The final GC-determined MFA-based CH4 prediction models estimate CH4 production, yield, and intensity with a root mean squared error of prediction of 35.7 g/d, 1.6 g/kg of DMI, and 1.6 g/kg of FPCM and with a concordance correlation coefficient of 0.72, 0.59, and 0.77, respectively. The final FTIR-based CH4 prediction models estimate CH4 production, yield, and intensity with a root mean squared error of prediction of 43.2 g/d, 1.9 g/kg of DMI, and 1.7 g/kg of FPCM and with a concordance correlation coefficient of 0.52, 0.40, and 0.72, respectively. The GC-determined MFA-based prediction models described a greater part of the observed variation in CH4 emission than did the FTIR-based models. The cross validation results indicate that all CH4 prediction models (both GC-determined MFA-based and FTIR-based models) are robust; the difference between the coefficient of determination and the coefficient of determination of cross validation ranged from 0.01 to 0.07. The results indicate that GC-determined MFA have a greater potential than FTIR spectra to estimate CH4 production, yield, and intensity. Both techniques hold potential but may not yet be ready to predict CH4 emission of dairy cows in practice. Additional CH4 measurements are needed to improve the accuracy and robustness of GC-determined MFA and FTIR spectra for CH4 prediction.

Short communication: The effect of linseed oil and DGAT1 K232A polymorphism on the methane emission prediction potential of milk fatty acids.

Several in vivo CH4 measurement techniques have been developed but are not suitable for precise and accurate large-scale measurements; hence, proxies for CH4 emissions in dairy cattle have been proposed, including the milk fatty acid (MFA) profile. The aim of the present study was to determine whether recently developed MFA-based prediction equations for CH4 emission are applicable to dairy cows with different diacylglycerol o-acyltransferase 1 (DGAT1) K232A polymorphism and fed diets with and without linseed oil. Data from a crossover design experiment were used, encompassing 2 dietary treatments (i.e., a control diet and a linseed oil diet, with a difference in dietary fat content of 22 g/kg of dry matter) and 24 lactating Holstein-Friesian cows (i.e., 12 cows with DGAT1 KK genotype and 12 cows with DGAT1 AA genotype). Enteric CH4 production was measured in climate respiration chambers and the MFA profile was analyzed using gas chromatography. Observed CH4 emissions were compared with CH4 emissions predicted by previously developed MFA-based CH4 prediction equations. The results indicate that different types of diets (i.e., with or without linseed oil), but not the DGAT1 K232A polymorphism, affect the ability of previously derived prediction equations to predict CH4 emission. However, the concordance correlation coefficient was smaller than or equal to 0.30 for both dietary treatments separately, both DGAT1 genotypes separately, and the complete data set. We therefore concluded that previously derived MFA-based CH4 prediction equations can neither accurately nor precisely predict CH4 emissions of dairy cows managed under strategies differing from those under which the original prediction equations were developed.

The relationship between milk metabolome and methane emission of Holstein Friesian dairy cows: Metabolic interpretation and prediction potential.

This study aimed to quantify the relationship between CH4 emission and fatty acids, volatile metabolites, and nonvolatile metabolites in milk of dairy cows fed forage-based diets. Data from 6 studies were used, including 27 dietary treatments and 123 individual observations from lactating Holstein-Friesian cows. These dietary treatments covered a large range of forage-based diets, with different qualities and proportions of grass silage and corn silage. Methane emission was measured in climate respiration chambers and expressed as production (g per day), yield (g per kg of dry matter intake; DMI), and intensity (g per kg of fat- and protein-corrected milk; FPCM). Milk samples were analyzed for fatty acids by gas chromatography, for volatile metabolites by gas chromatography-mass spectrometry, and for nonvolatile metabolites by nuclear magnetic resonance. Dry matter intake was 15.9 ± 1.90 kg/d (mean ± SD), FPCM yield was 25.2 ± 4.57 kg/d, CH4 production was 359 ± 51.1 g/d, CH4 yield was 22.6 ± 2.31 g/kg of DMI, and CH4 intensity was 14.5 ± 2.59 g/kg of FPCM. The results show that changes in individual milk metabolite concentrations can be related to the ruminal CH4 production pathways. Several of these relationships were diet driven, whereas some were partly dependent on FPCM yield. Next, prediction models were developed and subsequently evaluated based on root mean square error of prediction (RMSEP), concordance correlation coefficient (CCC) analysis, and random 10-fold cross-validation. The best models with milk fatty acids (in g/100 g of fatty acids; MFA) alone predicted CH4 production, yield, and intensity with a RMSEP of 34 g/d, 2.0 g/kg of DMI, and 1.7 g/kg of FPCM, and with a CCC of 0.67, 0.44, and 0.75, respectively. The CH4 prediction potential of both volatile metabolites alone and nonvolatile metabolites alone was low, regardless of the unit of CH4 emission, as evidenced by the low CCC values (<0.35). The best models combining the 3 types of metabolites as selection variables resulted in the inclusion of only MFA for CH4 production and CH4 yield. For CH4 intensity, MFA, volatile metabolites, and nonvolatile metabolites were included in the prediction model. This resulted in a small improvement in prediction potential (CCC of 0.80; RMSEP of 1.5 g/kg of FPCM) relative to MFA alone. These results indicate that volatile and nonvolatile metabolites in milk contain some information to increase our understanding of enteric CH4 production of dairy cows, but that it is not worthwhile to determine the volatile and nonvolatile metabolites in milk to estimate CH4 emission of dairy cows. We conclude that MFA have moderate potential to predict CH4 emission of dairy cattle fed forage-based diets, and that the models can aid in the effort to understand and mitigate CH4 emissions of dairy cows.

Linseed oil and DGAT1 K232A polymorphism: Effects on methane emission, energy and nitrogen metabolism, lactation performance, ruminal fermentation, and rumen microbial composition of Holstein-Friesian cows.

Complex interactions between rumen microbiota, cow genetics, and diet composition may exist. Therefore, the effect of linseed oil, DGAT1 K232A polymorphism (DGAT1), and the interaction between linseed oil and DGAT1 on CH4 and H2 emission, energy and N metabolism, lactation performance, ruminal fermentation, and rumen bacterial and archaeal composition was investigated. Twenty-four lactating Holstein-Friesian cows (i.e., 12 with DGAT1 KK genotype and 12 with DGAT1 AA genotype) were fed 2 diets in a crossover design: a control diet and a linseed oil diet (LSO) with a difference of 22 g/kg of dry matter (DM) in fat content between the 2 diets. Both diets consisted of 40% corn silage, 30% grass silage, and 30% concentrates (DM basis). Apparent digestibility, lactation performance, N and energy balance, and CH4 emission were measured in climate respiration chambers, and rumen fluid samples were collected using the oral stomach tube technique. No linseed oil by DGAT1 interactions were observed for digestibility, milk production and composition, energy and N balance, CH4 and H2 emissions, and rumen volatile fatty acid concentrations. The DGAT1 KK genotype was associated with a lower proportion of polyunsaturated fatty acids in milk fat, and with a higher milk fat and protein content, and proportion of saturated fatty acids in milk fat compared with the DGAT1 AA genotype, whereas the fat- and protein-corrected milk yield was unaffected by DGAT1. Also, DGAT1 did not affect nutrient digestibility, CH4 or H2 emission, ruminal fermentation or ruminal archaeal and bacterial concentrations. Rumen bacterial and archaeal composition was also unaffected in terms of the whole community, whereas at the genus level the relative abundances of some bacterial genera were found to be affected by DGAT1. The DGAT1 KK genotype was associated with a lower metabolizability (i.e., ratio of metabolizable to gross energy intake), and with a tendency for a lower milk N efficiency compared with the DGAT1 AA genotype. The LSO diet tended to decrease CH4 production (g/d) by 8%, and significantly decreased CH4 yield (g/kg of DM intake) by 6% and CH4 intensity (g/kg of fat- and protein-corrected milk) by 11%, but did not affect H2 emission. The LSO diet also decreased ruminal acetate molar proportion, the acetate to propionate ratio, and the archaea to bacteria ratio, whereas ruminal propionate molar proportion and milk N efficiency increased. Ruminal bacterial and archaeal composition tended to be affected by diet in terms of the whole community, with several bacterial genera found to be significantly affected by diet. These results indicate that DGAT1 does not affect enteric CH4 emission and production pathways, but that it does affect traits other than lactation characteristics, including metabolizability, N efficiency, and the relative abundance of Bifidobacterium. Additionally, linseed oil reduces CH4 emission independent of DGAT1 and affects the rumen microbiota and its fermentative activity.

Relationships between methane emission of Holstein Friesian dairy cows and fatty acids, volatile metabolites and non-volatile metabolites in milk.

This study investigated the relationships between methane (CH4) emission and fatty acids, volatile metabolites (V) and non-volatile metabolites (NV) in milk of dairy cows. Data from an experiment with 32 multiparous dairy cows and four diets were used. All diets had a roughage : concentrate ratio of 80 : 20 based on dry matter (DM). Roughage consisted of either 1000 g/kg DM grass silage (GS), 1000 g/kg DM maize silage (MS), or a mixture of both silages (667 g/kg DM GS and 333 g/kg DM MS; 333 g/kg DM GS and 677 g/kg DM MS). Methane emission was measured in climate respiration chambers and expressed as production (g/day), yield (g/kg dry matter intake; DMI) and intensity (g/kg fat- and protein-corrected milk; FPCM). Milk was sampled during the same days and analysed for fatty acids by gas chromatography, for V by gas chromatography-mass spectrometry, and for NV by nuclear magnetic resonance. Several models were obtained using a stepwise selection of (1) milk fatty acids (MFA), V or NV alone, and (2) the combination of MFA, V and NV, based on the minimum Akaike's information criterion statistic. Dry matter intake was 16.8±1.23 kg/day, FPCM yield was 25.0±3.14 kg/day, CH4 production was 406±37.0 g/day, CH4 yield was 24.1±1.87 g/kg DMI and CH4 intensity was 16.4±1.91 g/kg FPCM. The observed CH4 emissions were compared with the CH4 emissions predicted by the obtained models, based on concordance correlation coefficient (CCC) analysis. The best models with MFA alone predicted CH4 production, yield and intensity with a CCC of 0.80, 0.71 and 0.69, respectively. The best models combining the three types of metabolites included MFA and NV for CH4 production and CH4 yield, whereas for CH4 intensity MFA, NV and V were all included. These models predicted CH4 production, yield and intensity better with a higher CCC of 0.92, 0.78 and 0.93, respectively, and with increased accuracy (C b ) and precision (r). The results indicate that MFA alone have moderate to good potential to estimate CH4 emission, and furthermore that including V (CH4 intensity only) and NV increases the CH4 emission prediction potential. This holds particularly for the prediction model for CH4 intensity.

Milk metabolome relates enteric methane emission to milk synthesis and energy metabolism pathways.

Methane (CH4) emission of dairy cows contributes significantly to the carbon footprint of the dairy chain; therefore, a better understanding of CH4 formation is urgently needed. The present study explored the milk metabolome by gas chromatography-mass spectrometry (milk volatile metabolites) and nuclear magnetic resonance (milk nonvolatile metabolites) to better understand the biological pathways involved in CH4 emission in dairy cattle. Data were used from a randomized block design experiment with 32 multiparous Holstein-Friesian cows and 4 diets. All diets had a roughage:concentrate ratio of 80:20 (dry matter basis) and the roughage was grass silage (GS), corn silage (CS), or a mixture of both (67% GS, 33% CS; 33% GS, 67% CS). Methane emission was measured in climate respiration chambers and expressed as CH4 yield (per unit of dry matter intake) and CH4 intensity (per unit of fat- and protein-corrected milk; FPCM). No volatile or nonvolatile metabolite was positively related to CH4 yield, and acetone (measured as a volatile and as a nonvolatile metabolite) was negatively related to CH4 yield. The volatile metabolites 1-heptanol-decanol, 3-nonanone, ethanol, and tetrahydrofuran were positively related to CH4 intensity. None of the volatile metabolites was negatively related to CH4 intensity. The nonvolatile metabolites acetoacetate, creatinine, ethanol, formate, methylmalonate, and N-acetylsugar A were positively related to CH4 intensity, and uridine diphosphate (UDP)-hexose B and citrate were negatively related to CH4 intensity. Several volatile and nonvolatile metabolites that were correlated with CH4 intensity also were correlated with FPCM and not significantly related to CH4 intensity anymore when FPCM was included as covariate. This suggests that changes in these milk metabolites may be related to changes in milk yield or metabolic processes involved in milk synthesis. The UDP-hexose B was correlated with FPCM, whereas citrate was not. Both metabolites were still related to CH4 intensity when FPCM was included as covariate. The UDP-hexose B is an intermediate of lactose metabolism, and citrate is an important intermediate of Krebs cycle-related energy processes. Therefore, the negative correlation of UDP-hexose B and citrate with CH4 intensity may reflect a decrease in metabolic activity in the mammary gland. Our results suggest that an integrative approach including milk yield and composition, and dietary and animal traits will help to explain the biological metabolism of dairy cows in relation to methane CH4 emission.

Enteric methane production, rumen volatile fatty acid concentrations, and milk fatty acid composition in lactating Holstein-Friesian cows fed grass silage- or corn silage-based diets.

The objective of this study was to determine the effects of replacing grass silage (GS) with corn silage (CS) in dairy cow diets on enteric methane (CH4) production, rumen volatile fatty acid concentrations, and milk fatty acid (FA) composition. A completely randomized block design experiment was conducted with 32 multiparous lactating Holstein-Friesian cows. Four dietary treatments were used, all having a roughage-to-concentrate ratio of 80:20 based on dry matter (DM). The roughage consisted of either 100% GS, 67% GS and 33% CS, 33% GS and 67% CS, or 100% CS (all DM basis). Feed intake was restricted (95% of ad libitum DM intake) to avoid confounding effects of DM intake on CH4 production. Nutrient intake, apparent digestibility, milk production and composition, nitrogen (N) and energy balance, and CH4 production were measured during a 5-d period in climate respiration chambers after adaptation to the diet for 12 d. Increasing CS proportion linearly decreased neutral detergent fiber and crude protein intake and linearly increased starch intake. Milk production and milk fat content (on average 23.4 kg/d and 4.68%, respectively) were not affected by increasing CS inclusion, whereas milk protein content increased quadratically. Rumen variables were unaffected by increasing CS inclusion, except the molar proportion of butyrate, which increased linearly. Methane production (expressed as grams per day, grams per kilogram of fat- and protein-corrected milk, and as a percent of gross energy intake) decreased quadratically with increasing CS inclusion, and decreased linearly when expressed as grams of CH4 per kilogram of DM intake. In comparison with 100% GS, CH4 production was 11 and 8% reduced for the 100% CS diet when expressed per unit of DM intake and per unit fat- and protein-corrected milk, respectively. Nitrogen efficiency increased linearly with increased inclusion of CS. The concentration of trans C18:1 FA, C18:1 cis-12, and total CLA increased quadratically, and iso C16:0, C18:1 cis-13, and C18:2n-6 increased linearly, whereas the concentration of C15:0, iso C15:0, C17:0, and C18:3n-3 decreased linearly with increasing inclusion of CS. No differences were found in short- and medium-straight, even-chain FA concentrations, with the exception of C4:0 which increased linearly with increased inclusion of CS. Replacing GS with CS in a common forage-based diet for dairy cattle offers an effective strategy to decrease enteric CH4 production without negatively affecting dairy cow performance, although a critical level of starch in the diet seems to be needed.

Production and economic potentials of cattle in pasture-based systems of the western Amazon region of Brazil.

Our objectives were to evaluate strategies to improve productivity and economic returns from beef and dual-purpose cattle systems based on data collected on one dual-purpose (Bos taurus x Bos indicus) and two beef (Nellore) cattle farms in the western Amazon region of Brazil. Forage chemical composition and digestion rates of carbohydrate fractions of grazed Brachiaria decumbens and Brachiaria brizantha cv. Marandu grasses and Pueraria phaseoloides (tropical kudzu) legume were measured monthly during a 9-mo period from the end of one dry season to the end of the subsequent rainy season. Measurements of milk and growth responses to grazing these forages were used to predict animal productivity responses to dietary nutrient availability throughout an annual cycle. The ME available for gain in our simulations was always more limiting than metabolizable protein. The predicted ME available for gain was 0.50 kg/d for steers grazing B. brizantha and 0.40 kg/d for finishing steers grazing B. decumbens. Grasses contained more NDF and neutral detergent insoluble protein and less ME (P < 0.05) in the rainiest months than in the less rainy season, which resulted in 20% less predicted weight gain by growing steers (P < 0.05). Supplementation with sorghum grain was required to increase milk production and growth by 25 or 50% per animal, respectively, but this strategy was less profitable than current forage-only diets. Greater productivity of land and labor from higher stocking indicated greater net margins for beef production, but not for milk. This study suggested that more intensive beef production by judicious fertilization of grass-legume pastures and greater stocking density is the preferable strategy for owners of these cattle systems to improve economic returns under current conditions. It also might help decrease the motivation for additional forest clearing.

Trypanosoma brucei: ecto-phosphatase activity present on the surface of intact procyclic forms.

The results presented in this paper indicate that procyclic forms of Trypanosoma brucei possess a phosphatase activity detected in the external cell surface able to hydrolyze about 0.7 nmol.mg-1.min-1 p-nitrophenylphosphate. A faster rate of hydrolysis was observed when membrane-enriched fractions were used. This activity is weakly sensitive to 1 mM NaF, 10 mM tartrate and 10 mM levamizole but strongly inhibited by 0.1 mM vanadate. Inhibition by both NaF and vanadate have a competitive character. This phosphatase activity decreases by increasing the pH from 6.8 to 8.4, a pH range in which cell viability was maintained during at least 1 hour. In the membrane-enriched fractions this phosphatase activity showed to be an acid phosphatase. In addition, intact cells could catalyze the dephosphorylation of [32P]phosphocasein phosphorylated at serine and threonine residues.

Cowper's syringocele: an analysis of 15 consecutive cases.

Cystic dilatation of Cowper's gland ducts (Cowper's syringocele) is uncommon in children and is frequently asymptomatic, but it may cause urinary infection, haematuria, dysuria, and obstructive voiding symptoms. Fifteen consecutive children with syringocele aged 15 days to 15 years old are reported here. Only four patients with obstructive syringocele were successfully treated surgically; all the others were successfully treated conservatively. The true clinical significance of Cowper's syringocele lies in its potential to cause urethral obstruction. Careful clinical, radiological, endoscopic, and urodynamic evaluation is necessary in order to avoid unnecessary surgery.

Thapsigargin causes Ca2+ release and collapse of the membrane potential of Trypanosoma brucei mitochondria in situ and of isolated rat liver mitochondria.

Thapsigargin, previously reported to release Ca2+ from non-mitochondrial stores of different cell types, as well as nigericin, were found, when used at high concentrations, to release Ca2+ and collapse the membrane potential of Trypanosoma brucei bloodstream and procyclic trypomastigotes mitochondria in situ. At similarly high concentrations (> 10 microM), thapsigargin was also found to release Ca2+ and collapse the membrane potential of isolated rat liver mitochondria. These results indicate that care should be taken when attributing the effects of thapsigargin in intact cells to the specific inhibition of the sarcoplasmic and endoplasmic reticulum Ca(2+)-ATPase family of calcium pumps. In addition, we have found no evidence for an increase in intracellular Ca2+ by release of the ion from intracellular stores by nigericin, measuring changes in cytosolic Ca2+ by dual wavelength spectrofluorometry in fura-2-loaded T. brucei bloodstream trypomastigotes or measuring Ca2+ transport in digitonin-permeabilized cells.