Platelet lysate can support the development of a 3D-engineered skin for clinical application.

Safety concerns associated with foetal bovine serum (FBS) have restricted its translation into clinics. We hypothesised that platelet lysate (PL) can be utilised as a safe alternative to produce serum-free 3D-engineered skin. PL supported a short-term expansion of fibroblasts, with negligible replication-induced senescence and directed epidermal stratification. PL-expanded fibroblasts were phenotypically separated into three subpopulations of CD90+FAP+, CD90+FAP- and CD90-FAP+, based on CD90 (reticular marker) and FAP (papillary marker) expression profile. PL drove the expansion of the intermediate CD90+ FAP+ subpopulation in expense of reticular CD90+FAP-, which may be less fibrotic once grafted. The 3D-engineered skin cultured in PL was analysed by immunofluorescence using specific markers. Detection of ColIV and LMN-511 confirmed basement membrane. K10 confirmed near native differentiation pattern of neo-epidermis. CD29- and K5-positive interfollicular stem cells were also sustained. Transmission and scanning electron microscopies detailed the ultrastructure of the neo-dermis and neo-epidermis. To elucidate the underlying mechanism of the effect of PL on skin maturation, growth factor contents in PL were measured, and TGF-ß1 was identified as one of the most abundant. TGF-ß1 neutralising antibody reduced the number of Ki67-positive proliferative cells, suggesting TGF-ß1 plays a role in skin maturation. Moreover, the 3D-engineered skin was exposed to lucifer yellow on days 1, 3 and 5. Penetration of lucifer yellow into the skin was used as a semi-quantitative measure of improved barrier function over time. Our findings support the concept of PL as a safe and effective serum alternative for bioengineering skin for cell therapies.

Synthesis of Ruthenium-Doped TiO2 Nanotube Arrays for the Photocatalytic Degradation of Terasil Blue Dye.

The oxidation of terasil blue dye (TB) has been achieved by photocatalysis under UV light using well organized nanotubes of Ti/TiO2 doped with ruthenium (Ru/Ti/TiO2). Ru/Ti/TiO2 were fabricated by Ti anodization followed by impregnation with 0.04, 0.08 and 0.16 wt% of RuCl3 · xH2O or KRuO4 as precursor salts. Ti foil anodization was carried out in non-aqueous solutions and in aqueous solutions at pH 7 and pH 12.6. Scanning electron microscopy analyses revealed a morphology of well-defined, organized nanotubes of Ti/TiO2 and Ru/Ti/TiO2. The ruthenium was evenly distributed over Ti/TiO2 surface. Cluster formation was also observed for 0.16 wt% Ru loading. The X-ray diffraction spectra showed anatase and rutile phase in pure Ti/TiO2 and Ru/Ti/TiO2 nanotubes. Therefore, ruthenium doping did not modify the crystalline phase of Ti/TiO2. Raman spectra did not reveal the rutile phase of TiO2 in the composites which is attributed to the laser characteristics used. The optical band gap energies of the Ti/TiO2 and Ru/Ti/TiO2 nanotube arrays showed that ruthenium-doping on Ti/TiO2 had slight effect on the band-gap energy. A decrease in the band gap energy of pure Ti/TiO2 (Eg 2.88 eV) was observed by Ru loading of 0.04 wt% in the Ti/TiO2 nanotube arrays (Eg 2.60 eV). The kinetic rate constant of the photocatalytic oxidation of TB, using 0.08 wt% Ru load, was 1.56 and 1.39 times of that on Ti/TiO2 composite (0.01 min-1) synthetized at pH 12.6 impregnated in RuCl3 (0.0156 min-1) and KRuO4 (0.0139 min-1) solutions, respectively. Nevertheless, the highest rate constant value of TB degradation (87.8%) was 0.0172 min-1 using 0.16 wt% Ru loading on Ti/TiO2 composite synthetized at pH 7 impregnated in RuCl3; this represents 1.15 times of that on Ti/TiO2 composite (0.0150 min-1; 81.4% color removal) synthetized under the same conditions.

Comprehensive low-dose imaging of carotid and coronary arteries with a single-injection dual-source CT angiography protocol.

AIM: To assess the feasibility of a fast single-bolus combined carotid and coronary computed tomography angiography (CTA) protocol in asymptomatic patients. MATERIALS AND METHODS: Thirty-three consecutive patients (18 women and 15 men) with a median age of 61 ± 14 years old (range 37-87 years) with known or suspected atherosclerotic disease were enrolled in this prospective study. A single breath-hold, single biphasic injection protocol (50 ml at 3 ml/s, 50 ml at 5 ml/s, 50 ml saline flush at 5 ml/s) was used for combined CTA imaging of the supra-aortic (SAA) and coronary arteries (CA) on a 64-slice dual-source CT (DSCT) machine. Helical CTA acquisition of the SAA was followed by prospective electrocardiography (ECG)-triggered coronary CTA. Subjective (four-point scale) image quality and objective signal-to-noise (SNR) and contrast-to-noise (CNR) measurements were performed. Vascular disease was graded on a four-point scale (grade 1: absent; grade 2: mild, grade 3: moderate; grade 4: severe). The radiation dose was recorded for each patient. RESULTS: The average enhancement and subjective quality score of SAA and CA segments were 396 HU/358 HU and 1.2 ± 0.3/1.72 ± 0.4, respectively. The SNR was 27.1 ± 1.7 in the SAA and 21.6 ± 1.6 in the CA (p < 0.0001). The CNR was 18.1 ± 1.2 and 15.9 ± 1.8, respectively (p = 0.4). Four percent of SAA and 14% of CA segments (mostly due to peri-venous streak artefacts and small calibre, respectively) produced non-diagnostic images. SAA findings were as follows: 26/33 (79%) patients showed no disease and 6/33 (18%) had grade 2 and 1/33 (3%) had grade 3 disease. CA findings were as follows: 25/33 (76%) showed no disease and 6/33 (18%) patients had grade 2 and 2/33 (6%) had grade 3 disease. Five patients had disease in both districts. The average radiation dose for the combined CTA angiogram was 4.3 ± 0.6 mSv. CONCLUSION: A fast, low-dose combined DSCT angiography protocol appears technically feasible for imaging carotid and coronary atherosclerotic disease.

Innovative sugar-insecticide feeding bioassay for adult female Anopheles gambiae (Diptera: Culicidae).

The primary malaria vector in sub-Saharan Africa, Anopheles gambiae Giles (Diptera: Culicidae), is an anthropophagic and endophilic mosquito targeted for control with insecticides applied to interior resting surfaces and impregnated onto bed net materials. Effective malaria vector management involves monitoring the insecticide susceptibility of mosquito populations. Contemporary bioassays are based on mosquito contact with insecticide residues. We developed an innovative insecticide bioassay system that involves mosquito ingestion of a sugar-insecticide solution. The sucrose-permethrin solution in our bioassay system contained Trypan blue dye, creating a visual marker of insecticide ingestion in the mosquito's abdomen. Blue fecal spots deposited in the bioassay container provided further evidence of mosquito feeding. We used our bioassay to characterize the permethrin susceptibility of adult females of two strains of A. gambiae, one of which was susceptible and the other exhibited reduced susceptibility to permethrin. We compared the dose-response of both strains to permethrin in a forced-contact filter paper bioassay. Both assay approaches produced similar dose-dependent mortality, indicating that the feeding bioassay had appropriately characterized permethrin susceptibility for both mosquito strains.

Cardiac dual-source CT for the preoperative assessment of patients undergoing bariatric surgery.

AIM: To assess the diagnostic value of coronary dual-source computed tomography (DSCT) as a comprehensive, non-invasive tool in the preoperative cardiac evaluation of patients undergoing bariatric surgery. MATERIALS AND METHODS: Thirty consecutive obese [average body mass index (BMI): 45 ± 7.6, range: 35-59] patients (24 women; six men; median age: 52 ± 15 years) were enrolled in this institutional review board (IRB)-approved, Health Insurance Portability and Accountability Act of 1996 (HIPAA)-compliant prospective study. Calcium scoring (CaS) and electrocardiography (ECG)-gated images of the coronary arteries were obtained with a large body habitus protocol (120 kV; 430 mAs; 100 ml iodinated contrast medium at 7 ml/s injection rate) on a DSCT machine. Qualitative (four-point: 1 = excellent to 4 = not delineable) coronary segmental analysis, signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) measurements were performed. The presence and degree of vascular disease (four-grade scale: mild to severe) was correlated with CaS and cardiovascular (CV) risk stratification blood tests. In patients with severe stenosis (>70%), findings were compared with cardiac nuclear medicine imaging (single photon-emission computed tomography; SPECT) imaging. RESULTS: The average HR, enhancement, and quality score were 64 ± 7 beats/min, 288 ± 66 HU and 1.8 ± .5, respectively. Ninety-three percent (417/450) of the coronary segments were rated diagnostic. The SNRs and CNRs were 17 ± 9 and 12 ± 7 for the right coronary artery; 17 ± 8 and 12 ± 7 for the left main coronary artery; 16 ± 9 and 11 ± 7 for the left anterior descending coronary artery; and 15 ± 7 and 10 ± 6 for the left circumflex coronary artery. Ten of the 30 patients (33%) demonstrated coronary artery disease (CAD) of which two (6%) showed three-vessel disease. Four (13%) patients showed severe disease: in three of which the presence of significant stenosis was confirmed by SPECT and by catheter angiography in the fourth patient. Neither the CaS, nor the CV risk stratification tests showed significant correlation with presence or degree of CAD (p > 0.05). CONCLUSIONS: Coronary DSCT is a robust alternative imaging tool in the preoperative assessment of patients undergoing bariatric surgery.

Responses of Amblyomma americanum and Dermacentor variabilis to odorants that attract haematophagous insects.

Carbon dioxide (CO(2) ), 1-octen-3-ol, acetone, ammonium hydroxide, L-lactic-acid, dimethyl trisulphide and isobutyric acid were tested as attractants for two tick species, Amblyomma americanum and Dermacentor variabilis (Acari: Ixodidae), in dose-response bioassays using Y-tube olfactometers. Only CO(2) , acetone, 1-octen-3-ol and ammonium hydroxide elicited significant preferences from adult A. americanum, and only CO(2) was attractive to adult D. variabilis. Acetone, 1-octen-3-ol and ammonium hydroxide were separately evaluated at three doses against CO(2) (from dry ice) at a field site supporting a natural population of A. americanum nymphs and adults. Carbon dioxide consistently attracted the highest number of host-seeking ticks. However, for the first time, acetone, 1-octen-3-ol and ammonium hydroxide were shown to attract high numbers of A. americanum. Further research is needed to determine the utility of these semiochemicals as attractants in tick surveillance and area-wide management programmes.

Proof of concept for a novel insecticide bioassay based on sugar feeding by adult Aedes aegypti (Stegomyia aegypti).

Aedes aegypti L. (Stegomyia aegypti) (Diptera: Culicidae) is the principal vector of dengue and yellow fever viruses in tropical and subtropical regions of the world. Disease management is largely based on mosquito control achieved by insecticides applied to interior resting surfaces and through space sprays. Population monitoring to detect insecticide resistance is a significant component of integrated disease management programmes. We developed a bioassay method for assessing insecticide susceptibility based on the feeding activity of mosquitoes on plant sugars. Our prototype sugar-insecticide feeding bioassay system was composed of inexpensive, disposable components, contained minimal volumes of insecticide, and was compact and highly transportable. Individual mosquitoes were assayed in a plastic cup that contained a sucrose-permethrin solution. Trypan blue dye was added to create a visual marker in the mosquito's abdomen for ingested sucrose-permethrin solution. Blue faecal spots provided further evidence of solution ingestion. With the sugar-insecticide feeding bioassay, the permethrin susceptibility of Ae. aegypti females from two field-collected strains was characterized by probit analysis of dosage-response data. The field strains were also tested by forced contact of females with permethrin residues on filter paper. Dosage-response patterns were similar, indicating that the sugar-insecticide feeding bioassay had appropriately characterized the permethrin susceptibility of the two strains.

Novel field assays and the comparative repellency of BioUD(®) , DEET and permethrin against Amblyomma americanum.

Two new field bioassay methods were developed to compare the repellent activity of BioUD(®) (containing 7.75% 2-undecanone), 98.1% DEET and 0.5% permethrin against natural populations of nymphal Amblyomma americanum (L.) (Acari: Ixodidae). In a cloth sheet assay, pieces of material measuring 41 × 58 cm, separately treated with one of the test materials or the appropriate solvent carrier, were placed at random on the ground and baited with dry ice for 1 h. Mean numbers of ticks on repellent-treated sheets were significantly lower than on control sheets. There was no significant difference in the number of ticks collected between sheets treated with BioUD(®) and those treated with DEET. However, significantly fewer ticks were found on sheets treated with BioUD(®) or DEET than on permethrin-treated sheets. In a sock test, over-the-calf tube socks were treated with one of the test materials or the appropriate solvent carrier. Human volunteers wore a repellent-treated and a corresponding carrier-treated sock on either leg and walked randomly over an area of approximately 4000 m(2) for 15 min. Significantly fewer ticks were collected from socks treated with BioUD(®) or DEET than from socks treated with the carrier and there was no significant difference in repellency between these two agents. No difference in the mean number of ticks collected was found between permethrin-treated and corresponding carrier-treated socks. To examine the mechanism of repellency of BioUD(®) , a four-choice olfactometer was used to assess spatial repellency against adult A. americanum. As expected in the absence of a repellent, when all choices were represented by water-treated filter paper, ticks were equally distributed among the choices. When one choice consisted of BioUD(®) -treated filter paper and the remaining choices of water-treated paper, the distribution of ticks on the repellent-treated paper was significantly lower than might be expected to occur by chance, suggesting that repellency is at least partly achieved by an olfactory mechanism.

Growth performance, oxidative stress and immune status of newly weaned pigs fed peroxidized lipids with or without supplemental vitamin E or polyphenols.

BACKGROUND: This study evaluated the use of dietary vitamin E and polyphenols on growth, immune and oxidative status of weaned pigs fed peroxidized lipids. A total of 192 piglets (21 days of age and body weight of 6.62 ± 1.04 kg) were assigned within sex and weight blocks to a 2 × 3 factorial arrangement using 48 pens with 4 pigs per pen. Dietary treatments consisted of lipid peroxidation (6% edible soybean oil or 6% peroxidized soybean oil), and antioxidant supplementation (control diet containing 33 IU/kg DL-α-tocopheryl-acetate; control with 200 IU/kg additional dl-α-tocopheryl-acetate; or control with 400 mg/kg polyphenols). Pigs were fed in 2 phases for 14 and 21 days, respectively. RESULTS: Peroxidation of oil for 12 days at 80 °C with exposure to 50 L/min of air substantially increased peroxide values, anisidine value, hexanal, and 2,4-decadienal concentrations. Feeding peroxidized lipids decreased (P < 0.001) body weight (23.16 vs. 18.74 kg), daily gain (473 vs. 346 g/d), daily feed intake (658 vs. 535 g/d) and gain:feed ratio (719 vs. 647 g/kg). Lipid peroxidation decreased serum vitamin E (P < 0.001) and this decrease was larger on day 35 (1.82 vs. 0.81 mg/kg) than day 14 (1.95 vs. 1.38 mg/kg). Supplemental vitamin E, but not polyphenols, increased (P ≤ 0.002) serum vitamin E by 84% and 22% for control and peroxidized diets, respectively (interaction, P = 0.001). Serum malondialdehyde decreased (P < 0.001) with peroxidation on day 14, but not day 35 and protein carbonyl increased (P < 0.001) with peroxidation on day 35, but not day 14. Serum 8-hydroxydeoxyguanosine was not affected (P > 0.05). Total antioxidant capacity decreased with peroxidation (P < 0.001) and increased with vitamin E (P = 0.065) and polyphenols (P = 0.046) for the control oil diet only. Serum cytokine concentrations increased with feeding peroxidized lipids on day 35, but were not affected by antioxidant supplementation (P > 0.05). CONCLUSION: Feeding peroxidized lipids negatively impacted growth performance and antioxidant capacity of nursery pigs. Supplementation of vitamin E and polyphenols improved total antioxidant capacity, especially in pigs fed control diets, but did not restore growth performance.

Dual-energy CT angiography of pelvic and lower extremity arteries: dual-energy bone subtraction versus manual bone subtraction.

AIM: To analyse the effect of dual-energy bone subtraction (DEBS) on the image quality of peripheral computed tomography (CT) angiograms. MATERIALS AND METHODS: Twenty patients underwent dual-energy CT angiography of the pelvic and lower extremity arteries using commercially available equipment. Two different methods of image processing were employed for each CT angiographic dataset: (1) DEBS, and (2) manual bone subtraction (MBS). Effects on vessel visibility and artificial vessel alterations were compared. RESULTS: Bone removal, and the resultant visibility of vessel segments, were significantly better with DEBS than with MBS (p=0.011). The overall frequency of vessel-related alterations was lower in MBS compared with DEBS (p=0.001). Specifically, in the 249 vessel segments with calcified plaques, MBS generated fewer vessel alterations than DEBS (p<0.001). In the 309 vessel segments without calcified plaques, there was no difference in vessel alteration between the two techniques (p=0.22). CONCLUSION: DEBS facilitates bone removal in peripheral CT angiography, but generates more vessel alterations, particularly in the presence of calcified plaque.

Influence of gait on bone strength in turkeys with leg defects.

Leg problems have become more frequent in fast-growing turkeys. The objective of this study was to evaluate the effects of common leg defects on kinetic parameters of gait and biomechanical properties of bone. Nine hundred, day-old, male, Large White turkeys were raised in 48 floor pens. At 42 d of age, turkeys were divided into four categories of leg condition as determined by visual evaluation: Normal, Crooked toes, Shaky legs, and Valgus. Fifteen toms were selected from each group and trained to walk on a pressure sensitive walkway. Gait kinetic data were collected at 92, 115 and 144 d of age. At 145 d of age, turkeys were sacrificed and bones were collected and frozen until analysis. Morphological measurements of femur, tibia and tarsus-metatarsus were recorded. Bone mineral density (BMD) and content (BMC) were obtained using DEXA. Bone strength of tibias was evaluated using a four-point bending test and femurs with a torsion test. ANOVA was used to detect differences among groups, and Tukey's test used for mean separation. There were no differences in BW among different leg conditions. Gait parameters changed as turkeys aged, and age-group interactions were observed on peak vertical force (PVF), contact time, step length (SL) and bipedal cycle. No differences (P > 0.05) were detected in morphological measurements of femur or tibia. Relative asymmetry of femur length was lower (P < 0.05) in Normal and Valgus turkeys than in toms with Crooked toes. There were no differences (P > 0.05) among groups for femur BMD, BMC or strength. Tibia BMD and the area moment of inertia of turkeys with Crooked toes were lower (P < 0.05) than in toms with Valgus. With all four leg conditions, femur strength was positively correlated with PVF and negatively correlated with SL; BMD and BMC were correlated with tibia strength and gait kinetic parameters. In conclusion, only crooked toes caused consistent differences in gait patterns, bone properties and bone strength, but in general, gait kinetics was correlated with bone biomechanics in turkeys.

Gait parameters in four strains of turkeys and correlations with bone strength.

Locomotion problems in meat poultry have multifactorial etiology. A better understanding of normal gait and its influences on biomechanical aspects of leg bones among turkey genetic lines is important to prevent skeletal disorders and locomotion issues. The objective of this experiment was to determine the possible differences in gait kinetic and kinematic parameters of turkey strains and their effects on bone biomechanical properties. Four genetic lines, named A, B, C, and D, were obtained and raised in 48 floor pens with new pine shavings. Leg health issues were classified at 16 and 33 d of age. Fifteen turkeys from each strain with apparent normal legs and gait at 33 d of age were selected for gait analysis. These 15 turkeys were trained to walk on a pressure sensitive walkway and video was recorded to calculate articulation movements. These data also were analyzed to obtain kinetic and kinematic parameters of the gait cycle collected at 47, 84, 107, and 145 d of age. At 20 wk all turkeys were sacrificed, and legs were collected and frozen for analysis. Weights and morphologic measurements of the femur, tibia, and shank were recorded. Bone mineral density (BMD) and content (BMC) were obtained using DEXA. Femur and tibia strength were evaluated by a 4-point bending test and torsion test, respectively. Gait parameters changed as toms aged and some differences were observed among lines. Genetic lines differed on BMD, but not on BMC. Strain D had a higher BMD and smaller diaphyseal angle than strain C, characteristics that were correlated with stronger bones. Strain D also had the lowest incidence of leg problems while strain C had the highest. Furthermore, the D strain had a smaller vertical motion of the toe than strains C and B at 47 d and strain A at 145 d, indicating that the D strain had a more efficient gait. In summary, genetic strains differ significantly on gait parameters, which in turn impacts bone biomechanics.

Homogeneous and heterogenized Au(III) Schiff base-complexes as selective and general catalysts for self-coupling of aryl boronic acids.

A series of homogeneous and heterogenized gold metal complexes show high activity and selectivity for the homocoupling of a large variety of aryl boronic acids, being of general utility for the synthesis of C2-symmetric biaryls.

Validation of a liquid chromatography-mass spectrometry method to assess the metabolism of bupropion in rat everted gut sacs.

We have developed a rapid, sensitive and selective LC-MS method for the simultaneous assay of bupropion and its metabolite hydroxybupropion during its intestinal absorption, studied with the rat everted gut sac model. The method was validated in the concentration range of 1-15 microM (0.024-3.58 microg/mL) for bupropion and 0.005-1 microM (0.00127-0.25 microg/mL) for hydroxybupropion with 10 microL injected. Bupropion is used as a probe for the activity of the CYP2B6 isoenzyme of the P450 family of enzymes in man. Its major metabolite hydroxybupropion was found in the serosal media of the gut sac showing that the isoenzyme of the 2B group was active in the intestinal mucosa and metabolized bupropion during its passage across the mucosa. The metabolite was also quantified in the mucosal media indicating its ability to cross the apical membrane of the epithelial cells.

Validation of a liquid chromatography-mass spectrometry method to assess the metabolism of dextromethorphan in rat everted gut sacs.

A rapid, sensitive and selective liquid chromatography-mass spectrometry (LC-MS) method was developed for the simultaneous assay of dextromethorphan and its metabolites in tissue culture medium and its intestinal metabolism studied with the rat everted gut sac model. The method was validated in the concentration range of 0.1-2.5 microM (27.1 ng/mL-0.677 microg/mL) for dextromethorphan and 0.005-0.5 microM for dextrorphan and 3-methoxymorphinan (1.28 ng/mL-0.128 microg/mL) and 3-hydroxymorphinan (1.22 ng/mL-0.122 microg/mL). The limits of quantification (LOQ) were 0.0025 microM (12.5 fmoles, 3.4 pg, 5 microL injected) for dextromethorphan; 0.0025 microM for dextrorphan, 3-methoxymorphinan (24.9 fmoles, 6.4 pg injected), and 3-hydroxymorphinan (25.1 fmoles, 6.1 pg injected) with 10 microL injected. The detection of dextrorphan and 3-methoxymorphinan showed that both the P450 isoforms CYP3A and 2D were active in the intestinal mucosa and metabolised dextromethorphan during its passage across the mucosa.

N-acetyl-beta-D-hexosaminidase from Trichomonas vaginalis: substrate specificity and activity of inhibitors.

Among chitinolytic activities previously described in Trichomonas vaginalis, N-acetyl-beta-D-hexosaminidase (NAHase) was the enzyme system expressing the highest level of specific activity. We report here some biochemical characteristics of NAHase purified from T. vaginalis. We found at first that the use of 4-methylumbellifferyl-substrate was responsible for a substrate affinity for the enzyme, about 1000-fold higher than those when using p-nitrophenyl-substrates (PNP). Whereas the optimum pH was 7.0 using PNP-substrate, it was at 4.5 using 4-methylumbelliferyl-substrate. Four different substrates were compared for their action on T. vaginalis NAHase and we have found that N-acetyl-beta-D-glucosaminide substrate was the most specific. DTT had no effect on enzyme activity suggesting that thiol group are not involved at the catalytic site. The use of previously described inhibitors showed a positive correlation between trichomonacidal activity and NAHase inhibition.

Development of prediction equations to estimate the apparent digestible energy content of lipids when fed to lactating sows.

Two studies were conducted 1) to determine the effects of free fatty acid (FFA) concentrations and the degree of saturation of lipids (unsaturated to saturated fatty acids ratio [U:S]) on apparent total tract digestibility (ATTD) and DE content of lipids and 2) to derive prediction equations to estimate the DE content of lipids when added to lactating sow diets. In Exp. 1, 85 lactating sows were assigned randomly to a 4 × 5 factorial arrangement of treatments plus a control diet with no added lipid. Factors included 1) FFA concentrations of 0, 18, 36, and 54% and 2) U:S of 2.0, 2.8, 3.5, 4.2, and 4.9. Diets were corn-soybean meal based and lipid was supplemented at 6%. Concentrations of FFA and U:S were obtained by blending 4 lipid sources: choice white grease (CWG; FFA = 0.3% and U:S = 2.0), soybean oil (FFA = 0.1% and U:S = 5.5), CWG acid oil (FFA = 57.8% and U:S = 2.1), and soybean-cottonseed acid oil (FFA = 67.5% and U:S = 3.8). Titanium dioxide was added to diets (0.5%) as a digestibility marker. Treatments started on d 4 of lactation and fecal samples were collected after 6 d of adaptation to diets on a daily basis from d 10 to 13. The ATTD of added lipid and DE content of lipids were negatively affected (linear, < 0.001) with increasing FFA concentrations, but negative effects were less pronounced with increasing U:S (interaction, < 0.05). Coefficients of ATTD for the added lipid and DE content of lipids increased with increasing U:S (quadratic, = 0.001), but these improvements were less pronounced when the FFA concentration was less than 36%. Digestible energy content of added lipid was described by DE (kcal/kg) = [8,381 - (80.6 × FFA) + (0.4 × FFA) + (248.8 × U:S) - (28.1 × U:S) + (12.8 × FFA × U:S)] ( = 0.74). This prediction equation was validated in Exp. 2, in which 24 lactating sows were fed diets supplemented with 6% of either an animal-vegetable blend (A-V; FFA = 14.5% and U:S = 2.3) or CWG (FFA = 3.7% and U:S = 1.5) plus a control diet with no added lipids. Digestible energy content of A-V (8,317 and 8,127 kcal/kg for measured and predicted values, respectively) and CWG (8,452 and 8,468 kcal/kg for measured and predicted values, respectively) were accurately estimated using the proposed equation. The proposed equation involving FFA concentration and U:S resulted in highly accurate estimations of DE content (relative error, +0.2 to -2.3%) of commercial sources of lipids for lactating sows.

Population-based study of the incidence of Shigella diarrhea and causative serotypes in Santiago, Chile.

BACKGROUND: Shigella is an important cause of diarrheal disease in children in developing countries. The increasing prevalence of antibiotic-resistant strains has stimulated interest in the use of multivalent Shigella vaccines. Because Shigella vaccines under development are based on eliciting immunity to O antigens, monitoring the distribution of serotypes in defined target populations is critical. We initiated health center-based surveillance in a poor semirural community in Colina, Santiago (7489 children <60 months of age) to determine the age-specific incidence of Shigella disease and the responsible serotypes. FINDINGS: Surveillance was maintained at the 2 health centers during warm seasons (November 1 through April 30) for 4 successive years (1994 to 1998). Shigella was recovered from 54 of 243 cases of dysentery (22%) and from 215 of 3966 cases of nondysenteric diarrhea (5.4%) (P < 0.001). The peak mean annual incidence of shigellosis occurred among children 12 to 47 months of age (9.0 to 12.6 cases/10(3) children), although the incidence in infants (5.2/10(3)) and children 48 to 59 months of age (6.2/10(3)) was also substantial. During the 1995 through 1996 season, an age-matched healthy control was cultured for every child <60 months of age with diarrhea. Shigella isolation from cases (34 of 576, 5.9%) was >8-fold higher than controls (4 of 576, 0.7%) (P < 0.01). Four serotypes, Shigella sonnei (45%), Shigella flexneri 2b (19%), S. flexneri 2a (14%) and S. flexneri 6 (11%), accounted for 89% of all cases. INTERPRETATION: Shigella remains an important pediatric pathogen in Santiago. The serotype distribution from Colina, which closely resembles data from a population-based surveillance study in Santiago in the mid-1980s, demonstrates a remarkable degree of serotype stability in Santiago during a 15-year period.

Liquid chromatographic-mass spectrometric method to assess cytochrome P450-mediated metabolism of testosterone by rat everted gut sacs.

A rapid, sensitive and specific method was developed for the simultaneous assay of testosterone, androstenedione and 6beta-hydroxytestosterone (6beta-OHT) in the TC199 tissue culture medium used in intestinal drug metabolism studies with the rat everted gut sac model. An electrospray LC-MS method was validated in the concentration range of 0.025-9.5 microM (7.2 ng-2.7 microg/mL) for testosterone and androstenedione and 0.01-4 microM (3 ng-1.2 microg/mL) for 6beta-hydroxytestosterone. The limits of quantification (LOQ) with an injection volume of 10 microL were 0.0005 microM (4.9 fmol, 1.4 pg injected), 0.004 microM (0.04 pmol, 11.4 pg injected) and 0.03 microM (0.3 pmol, 91 pg injected), respectively. The method also detected the other testosterone metabolites, the 16alpha-, 16beta-, 2beta- and 2alpha-hydroxytestosterones and was then used to study the metabolism of testosterone during its absorption by rat intestine in vitro, using everted gut sacs.