Standardized ileal amino acid digestibility of expeller-extracted canola meal subjected to different processing conditions for starter and grower broiler chickens.

Six expeller-extracted canola meal (ECM) samples produced under different seed conditioning temperatures (90, 95, or 100°C) and screw torques in the second press (low or high) were evaluated in a 3 × 2 factorial arrangement of treatments to determine the effect of processing on standardized ileal amino acid digestibility (SID) in 10 and 24-day-old broilers. A nitrogen-free diet was fed to determine ileal endogenous amino acid flow. Each diet was fed for 5 d to 6 replicate cages of 10 or 7 chicks, and ileal samples were collected at 10 and 24 d, respectively. The endogenous flow (mg/kg DM intake) of CP and all amino acids except Cys decreased (P < 0.01) with age. Conditioning temperature by screw torque interactions were detected (P < 0.05) for apparent ileal digestibility (AID) of CP, Arg, Ile, Lys, Phe, Thr, Ala, Asp, Glu, Pro ,and Ser at d 10, and the same trend was observed at d 24 except for Ile, Phe, Ala, and Pro (P > 0.05). Meals processed under medium conditioning temperature (95°C) at either low or high screw torque had the greatest (P < 0.05) AID of CP and total amino acids. The SID values followed a similar pattern as AID at both ages. When corrected for endogenous losses, the average AID of total amino acids improved approximately 3.5 and 2.0 units, at 10 and 24 d, respectively. The AID and SID values increased (P < 0.05) with age for most AA, but the effect of age was not consistent between ECMs. A negative correlation was detected between NDF and neutral detergent-insoluble nitrogen (NDIN) content of the meals and SID values of Lys at d 24 (r = -0.79, r = -0.76; P = 0.001, respectively). Processing conditions affected CP and amino acid digestibility, likely because of alterations to the chemical composition of ECM and formation of indigestible complexes of amino acids with fiber. The AID and SID values increased with age independent of meal processing conditions.

Apparent metabolizable energy value of expeller-extracted canola meal subjected to different processing conditions for growing broiler chickens.

The objective of this study was to investigate the effect of processing conditions and chemical composition on ileal digestible energy (IDE), AME, and AMEn of 6 expeller-extracted canola meal (ECM) samples subjected to conditioning temperature at 90, 95, or 100°C and high or low screw torque over the second presses in a 3 × 2 factorial arrangement. The ECM samples were incorporated into a corn-soybean meal reference diet at 30% by replacing energy-yielding ingredients. A total of 210 one-day-old male broiler chicks (Ross 308) were fed common starter and grower diets until d 18, and then assigned to 7 experimental diets replicated 6 times, with 5 chicks per cage. After a 5-d diet acclimation period from d 18 to 22, excreta was collected for 72 h. The difference method was used to determine AME, which was corrected to zero N balance to obtain AMEn. Medium seed conditioning temperature resulted in the highest IDE, AME, and AMEn compared with low or high temperature, and high screw torque resulted in higher energy utilization compared with low torque (P < 0.001). There was also an interaction (P < 0.001) between conditioning temperature and screw torque. For ECM subjected to low or medium conditioning temperature at low screw torque, IDE, AME, and AMEn values ranging from 2,137 to 2,705, 2,089 to 2,655, and 1,977 to 2,482 kcal/kg of DM, respectively, were obtained. The mean AMEn values were 2,260 kcal/kg of DM, indicating a 7% reduction compared with AME values. The AMEn values were negatively correlated with neutral detergent fiber (NDF; r = -0.93; P = 0.001) and NDIN (r = -0.87; P = 0.001). Stepwise regression to predict AMEn value resulted in the following equation: AMEn (kcal/kg of DM) = 3,397.8 + (-100.1 × NDF %) + (279.5 × ash %) + (-33.8 × ADF %) (R² = 0.91; SE = 61.9; P = 0.001). These results indicate that AMEn values vary markedly among ECM samples, and chemical constituents, especially the fiber components, may have a considerable effect on AMEn value.

Evaluation of high dietary inclusion of distillers dried grains with solubles and supplementation of protease and xylanase in the diets of broiler chickens under necrotic enteritis challenge.

A 2 × 2 × 2 factorial experiment was conducted to investigate the effect of a high level of sorghum distillers dried grains with solubles (DDGS; 20%), with or without a combination of protease and xylanase in broiler chickens, under a necrotic enteritis disease challenge. A total of 576 male broiler chicks were randomly assigned to 8 experimental treatments, each replicated 6 times, with 12 birds per replicate for 35 d. Oral inoculation of the challenged group with Eimeria spp. occurred on d 9, followed by 3 consecutive inoculations of Clostridium perfringens from d 14 through 16. The disease challenge and DDGS inclusion significantly (P < 0.01) interacted, depressing BW gain and feed conversion ratio only in wk 3. Disease challenge adversely influenced (P < 0.01) BW gain and feed conversion ratio of the birds in the third week and across the 35-d study. Over the last 2 wk and across the 35-d trial, the interaction between DDGS and enzyme supplementation showed a tendency (P = 0.09) to gain more BW in birds regardless of the disease challenge. Inclusion of 20% DDGS markedly (P < 0.01) interacted with disease challenge, accelerating the proliferation of C. perfringens in the ceca at d 17. Inoculation of birds with C. perfringens resulted in higher (P < 0.01) counts of C. perfringens in both ileal and cecal contents. The necrotic enteritis-related lesions (d 17) were more severe (P < 0.05) in the intestine of infected birds fed DDGS diets than in birds fed the control diet. Incorporation of DDGS to the diets improved (P < 0.01) the IgA and IgG titer at d 13 but interacted with the disease challenge, reducing the concentration of IgA at d 21 and IgM at d 35 in the infected birds. In conclusion, incorporating a high level of DDGS in the diet of broiler chickens may increase susceptibility to necrotic enteritis. Supplementation of enzymes did not reveal significant mitigation effect in infected birds but helped the birds fed DDGS to maintain feed intake and BW gain.

CLASP2 facilitates dynamic actin filament organization along the microtubule lattice.

Coordination between the microtubule and actin networks is essential for cell motility, neuronal growth cone guidance, and wound healing. Members of the CLASP (cytoplasmic linker-associated protein) family of proteins have been implicated in the cytoskeletal cross-talk between microtubules and actin networks; however, the molecular mechanisms underlying the role of CLASP in cytoskeletal coordination are unclear. Here, we investigate CLASP2α's cross-linking function with microtubules and F-actin. Our results demonstrate that CLASP2α cross-links F-actin to the microtubule lattice in vitro. We find that the cross-linking ability is retained by L-TOG2-S, a minimal construct containing the TOG2 domain and serine-arginine-rich region of CLASP2α. Furthermore, CLASP2α promotes the accumulation of multiple actin filaments along the microtubule, supporting up to 11 F-actin landing events on a single microtubule lattice region. CLASP2α also facilitates the dynamic organization of polymerizing actin filaments templated by the microtubule network, with F-actin forming bridges between individual microtubules. Finally, we find that depletion of CLASPs in vascular smooth muscle cells results in disorganized actin fibers and reduced coalignment of actin fibers with microtubules, suggesting that CLASP and microtubules contribute to higher-order actin structures. Taken together, our results indicate that CLASP2α can directly cross-link F-actin to microtubules and that this microtubule-CLASP-actin interaction may influence overall cytoskeletal organization in cells.

Kaposi's sarcoma-associated herpesvirus protein ORF75 among HIV-1 patients in Kenya.

BACKGROUND: Histology is used to identify Kaposi's sarcoma (KS) in countries with low resources to fund healthcare costs. Approximately 95% of KS cases can be detected using a polymerase chain reaction. OBJECTIVE: To determine the presence of the open reading frame 75 (ORF75) gene associated with Kaposi's sarcoma herpes virus among HIV-1/AIDS patients and to describe morphological presentations of KS. METHODS: This was a retrospective, descriptive study of archived tissue blocks collected from 2013 to 2016. Haematoxylin and eosin staining was used to identify KS. Deoxyribonucleic acid from archived tissue blocks was extracted and a nested polymerase chain reaction was used to detect the ORF75 gene. RESULTS: All 81 cases in this study had been diagnosed as HIV-1 positive, of which 68 had hallmark features of KS in the histology report and 13 had features suggestive of KS ('KS-like'). Microscopic identification of KS by haematoxylin and eosin staining was considered a significant indicator of KS herpes virus ORF75 gene positivity (p = 0.002). The ORF75 gene was detected in 60.5% (49/81) of tissue blocks; 27.2% were men (22/81) and 33.3% were women (27/81). The ORF75 gene was observed to be present in up to 15.4% (2/13) of the cases reported to have KS-like features. CONCLUSION: Following the initial diagnosis of KS by histology, the ORF75 gene was fur-ther detected from both cases that had hallmark features of KS as well as among cases with KS-like fea-tures.

Benzo[a]pyrene metabolism in organ cultures of human endometrium.

Twenty-six specimens of normal human endometrium were obtained for studies of [3H]benzo[alpha]pyrene metabolism in short-term organ culture. The tissues were capable of converting benzo[alpha]pyrene (BP) to oxygenated derivatives which co-chromatographed with dihydrodiols, quinones and monohydroxy derivatives of BP. A sulfate conjugate of a monohydroxy BP was also found among the acetone/ethyl acetate soluble products. When the acetone/ethyl acetate soluble metabolites were analyzed using high-pressure liquid chromatography (HPLC), the proportion of metabolites were found to vary greatly from specimen to specimen. Eight women smoked cigarettes but the BP metabolite profiles from organ cultures of endometrial tissue from these women did not differ from those who did not smoke. The average value for sulfate conjugates of monohydroxy BP was significantly lower in endometrial tissue from three postmenopausal women (ages 49, 57 and 58) than among premenopausal women. Specimens from premenopausal women which had been in organ culture for 2 weeks prior to the addition of [3H] BP produced metabolite profiles similar to those of specimens from postmenopausal women with a greatly reduced proportion of BP-phenol sulfate conjugates. These data suggest that the absence of hormonal stimulus (i.e., advanced age or conditions of organ culture) may affect the metabolism of BP in human endometrial tissue.

Palliation of pancreatic cancer using electrolytic ablation.

BACKGROUND: Inoperable pancreatic cancer has a dismal prognosis. Palliation involves either stenting or surgical bypass. Stenting does not relieve gastric outlet obstruction, and surgical bypass is a major procedure. A minimally invasive procedure is needed that relieves both gastric outlet and biliary obstruction, with the potential for relieving pain. METHODS: In an experimental model, pancreatic electrolysis was investigated. The pancreatic duct was cannulated via a transduodenal approach with an electrode catheter. In 6 animals an electrolytic "lesion" was created using a direct current generator. Six animals were controls. The local and systemic effects of electrolysis were assessed using histological and biochemical parameters. RESULTS: The pancreatic duct was cannulated in all animals and treatment was uneventful. Electrolytic lesions comprised a central area of necrosis with a sharp demarcation between necrotic and viable pancreas. All animals developed transient hyperamylasemia after electrolysis. There was no significant difference between treatment and controls. Importantly, no animal had clinical, biochemical, or histological evidence of pancreatitis. CONCLUSIONS: This experimental study suggested that electrolytic palliation of inoperable pancreatic cancer via the gastrointestinal tract is potentially safe. In patients, this treatment could be performed during endoscopic retrograde cholangiopancreatography and may have therapeutic advantages when compared to stenting or biliary bypass.

Anionic salts and dietary 25-hydroxyvitamin D stimulate calcium availability in steers.

The influence of feeds containing varying dietary cation-anion differences (DCADs) with and without supplements of 25-hydroxyvitamin D (25(OH)D) on urine pH and excretion of macro minerals was determined in fistulated crossbred steers (mean live weight 315 ± 45 kg). A basal forage diet comprising lucerne hay and wheat chaff was used, to which varying quantities of MgCl(2) or K(2)CO(3) were added to achieve four levels of DCAD: -300, 50, 150 or 250 mEq/kg dry matter (DM). Steers were allocated to one of six treatments, one treatment for each diet and a further treatment for both the 50 and 150 mEq/kg DCAD diets, which were supplemented with 25(OH)D at a rate of 3 mg/steer per day. Urine pH from steers offered the diets comprising DCADs of 50, 150 and 250 mEq/kg ranging from 8.3 to 8.8. In treatments not containing 25(OH)D with DCADs of 50 to 250 mEq/kg, there were no significant differences in urine pH or Ca excretion. However, steers offered the diet with a DCAD of -300 mEq/kg DM produced urine with a significantly lower pH (6.5 to 7.5). Daily output of Ca in urine was also significantly higher from steers given this diet. Supplementation with 25(OH)D significantly increased urinary Ca excretion from steers offered diets of DCADs 50 and 150 mEq/kg DM. Estimates of daily urinary Ca excretion, calculated using the ratio of creatinine to Ca in 'spot' urine samples, were less variable than those based on total collection (residual mean square of 0.54 and 0.63, respectively).

The measurement of cytosolic estrogen receptors in human endometrial tissue and organ cultures.

PIP: This paper describes a method for determining the number of cytosolic estrogen receptors in small quantities of human endometrial tissue. The method is a modification of Puca et al.'s gel filtration technique developed for measurement of estrogen receptors in whole calf uteri. Sucrose gradient ultracentrifugation was used to determine sedimentation coefficients in the estrogen binding proteins of endometrial cytosol. Scatchard and double reciprocal analyses were used to determine graphically the total number of binding sites and dissociation constants. Estrogen receptor levels were measured in 22 patients at various points in the menstrual cycle. The highest receptor levels among the patients were observed in the endometrium histologically determined to be in the middle of the proliferative phase of the menstrual cycle. The quantity of the receptors decreased dramatically during the last part of the proliferative phase, the reduced levels persisting during the secretory phase. Estrogen receptors were also found in organ cultures of human endometrium maintained in vitro for up to 67 days. Estrogen receptors were found to have a sedimentation coefficient of approximately 8S, a dissociation constant of approximately 1 x 10 -9 molar, and a binding capacity ranging from 2.3 x 10 -13 to 8 x 10 -15 mol/mg of protein. This study shows that receptor assays can be performed with 100-200 mg of tissues using 10 mM triated hydrogen-estradiol binding and gel filtration method. The assay reported here may also be useful in measuring estrogen receptors of small clinical specimens, and in analyzing needle biopsy specimens or currettings.^ieng