Effect of the level of cholecalciferol supplementation of broiler diets on the performance and tibial dyschondroplasia.

A study was conducted to evaluate four different cholecalciferol levels (NRC; modified), using diets supplemented with 200 (control), 1500, 2500 or 3500 IU/kg of cholecalciferol (VIT-D3). Each treatment was assigned to 3 pens of 17 broiler chicks of a commercial strain grown in an open-sided house with sidewall curtains. At 21 and 42 days, BW and feed conversion (FCR) were determined. At 42 days, five birds per pen were slaughtered to evaluate tibia and toe ash of the right leg, and incidence and severity of tibial dyschondroplasia (TD) of the left tibia and also measured dressing percentage and breast meat yield. Serum calcium and phosphorus concentrations were also determined. Haemagglutination inhibition antibody titre against Newcastle disease virus and lymphoid organs weight/body weight ratio were also determined. At both 21 and 42 days, the BW of birds fed 1500 IU/kg to 3500 IU/kg of VIT-D3 was significantly greater than birds fed 200 IU/kg. Similarly, better FCR was observed in birds those fed diets of high level of VIT-D3. No significant difference was observed for mortality at any age. Better dressing percentage and breast meat yield were noted in birds fed diets containing 2500 or 3500 IU/kg VIT-D3. Both tibia and toe ash contents were increased (p < 0.05) progressively with increased concentrations of cholecalciferol in feed. The incidence of TD (percentage of birds having TD scores greater than zero) was significantly (p < 0.05) influenced by level of 3500 IU VIT-D3/kg at 42 days. The severity of TD in birds fed diets containing 200 IU/kg VIT-D3 was apparently higher than birds fed diets with higher levels of VIT-D3. Concentrations of calcium and phosphorus minerals in the serum increased progressively with the high level of VIT-D3 supplementation to birds at both 21 and 42 days of age. Feeding levels of 1500 or 3500 IU of vitamin D3 did positively affect the immune system within the parameters measured. It may be concluded that performance, bone mineralization, blood chemistry and immunity against disease in broilers could be maintained when supplementing high level of VIT-D3 incorporated in broiler diets.

PF-114, a potent and selective inhibitor of native and mutated BCR/ABL is active against Philadelphia chromosome-positive (Ph+) leukemias harboring the T315I mutation.

Targeting BCR/ABL with tyrosine kinase inhibitors (TKIs) is a proven concept for the treatment of Philadelphia chromosome-positive (Ph+) leukemias. Resistance attributable to either kinase mutations in BCR/ABL or nonmutational mechanisms remains the major clinical challenge. With the exception of ponatinib, all approved TKIs are unable to inhibit the 'gatekeeper' mutation T315I. However, a broad spectrum of kinase inhibition increases the off-target effects of TKIs and may be responsible for cardiovascular issues of ponatinib. Thus, there is a need for more selective options for the treatment of resistant Ph+ leukemias. PF-114 is a novel TKI developed with the specifications of (i) targeting T315I and other resistance mutations in BCR/ABL; (ii) achieving a high selectivity to improve safety; and (iii) overcoming nonmutational resistance in Ph+ leukemias. PF-114 inhibited BCR/ABL and clinically important mutants including T315I at nanomolar concentrations. It suppressed primary Ph+ acute lymphatic leukemia-derived long-term cultures that either displayed nonmutational resistance or harbor the T315I. In BCR/ABL- or BCR/ABL-T315I-driven murine leukemia as well as in xenograft models of primary Ph+ leukemia harboring the T315I, PF-114 significantly prolonged survival to a similar extent as ponatinib. Our work supports clinical evaluation of PF-114 for the treatment of resistant Ph+ leukemia.

Oligomerization inhibition, combined with allosteric inhibition, abrogates the transformation potential of T315I-positive BCR/ABL.

The t(9;22) translocation leads to the formation of the chimeric bcr/abl fusion gene, which encodes the BCR/ABL fusion protein. In contrast to its physiological counterpart c-ABL, the BCR/ABL kinase is constitutively activated, inducing the leukemic phenotype. The N-terminus of c-ABL (Cap region) contributes to the regulation of its kinase function. It is myristoylated, and the myristate residue binds to a hydrophobic pocket in the kinase domain known as the myristoyl-binding pocket in a process called 'capping', which results in an auto-inhibited conformation. Because the cap region is replaced by the N-terminus of BCR, the BCR/ABL 'escapes' this auto-inhibition. Allosteric inhibition by myristate 'mimics', such as GNF-2, is able to inhibit unmutated BCR/ABL, but not the BCR/ABL that harbors the 'gatekeeper' mutation T315I. In this study, we analyzed the possibility of increasing the efficacy of allosteric inhibition by blocking BCR/ABL oligomerization. We showed that inhibition of oligomerization was able to not only increase the efficacy of GNF-2 on unmutated BCR/ABL, but also overcome the resistance of BCR/ABL-T315I to allosteric inhibition. These results strongly suggest that the response to allosteric inhibition by GNF-2 is inversely related to the degree of oligomerization of BCR/ABL. In summary, our observations establish a new approach for the molecular targeting of BCR/ABL and its resistant mutants represented by the combination of oligomerization and allosteric inhibitors.

The gatekeeper mutation T315I confers resistance against small molecules by increasing or restoring the ABL-kinase activity accompanied by aberrant transphosphorylation of endogenous BCR, even in loss-of-function mutants of BCR/ABL.

In Philadelphia chromosome-positive (Ph+) leukemia BCR/ABL induces the leukemic phenotype. Targeted inhibition of BCR/ABL by kinase inhibitors leads to complete remission. However, patients with advanced Ph+ leukemia relapse and acquire resistance, mainly due to point mutations in BCR/ABL. The 'gatekeeper mutation' T315I is responsible for a general resistance to small molecules. It seems not only to decrease the affinity for kinase inhibitors, but to also confer additional features to the leukemogenic potential of BCR/ABL. To determine the role of T315I in resistance to the inhibition of oligomerization and in the leukemogenic potential of BCR/ABL, we investigated its influence on loss-of-function mutants with regard to the capacity to mediate factor independence. Here, we show that T315I (i) requires autophosphorylation at tyrosine 177 in the BCR-portion to mediate resistance against the inhibition of oligomerization; (ii) restores the capacity to mediate factor-independent growth of loss-of-function mutants due to an increase in or activation of ABL-kinase; (iii) leads to phosphorylation of endogenous BCR, suggesting aberrant substrate activation by BCR/ABL harboring the T315I mutation. These data show that T315I confers additional leukemogenic activity to BCR/ABL, which might explain the clinical behavior of patients with BCR/ABL-T315I-positive blasts.

Time of lay in hens exposed to intermittent lighting.

1. Two short-term trials are described in which laying hens were exposed to 8 h light followed by 8 one-min pulses of light at hourly intervals followed by 8 h darkness (8L:8i:8D). The effect of varying the intensity of illumination during the one-min pulses and the effect of placing the intermittent lighting before the 8 h photoperiod (8i:8L:8D), were studied. 2. Normal egg production was maintained by the 8L:8i:8D system when the light pulses were at 20 lux, but not at 5 lux. This suggests a minimum threshold for illumination with short light pulses higher than that needed for continuous lighting. 3. Time of lay under 8L:8i:8D was the same as with 8L:16D in relation to the beginning and ending of the 8 h main photoperiod, but with 8i:8L:8D mean time of lay was 2 to 3 h earlier. Thus the hourly pulses caused a phase advance when placed before the normal photoperiod but did not cause a phase delay when placed after the normal photoperiod.

Timing of oviposition in mixed systems using bright light, dim light and darkness.

1. Two experiments were conducted to examine time of oviposition for hens exposed to continuous dim lighting, to dim lighting alternating with bright lighting in a 24 h cycle or to a mixed system using bright light, dim light and darkness. 2. Under continuous dim lighting (0.3 lux), the pattern of ovipositions was the same as that reported previously for constant darkness, more eggs being laid around midnight than around noon. 3. With alternating bright and dim phases, mean time of lay was approximately 16 h after the transition from bright to dim lighting, which was 3 h earlier than under the corresponding cycle of light and dark. This phase advance was the same whether the bright:dim ratio was 16:1 or 160:1. 4. Dim lighting (1.25 lux) preceded by a period of normal lighting (5 lux or 50 lux) and followed by 8 h darkness was treated as part of the photoperiod. 5. It is concluded that, when there is no darkness, a period of dim lighting is treated as darkness, provided the contrast between bright and dim phases is sufficient. However, when darkness, dim light and bright light are all included in a cycle, the dim light is treated as part of the photoperiod, even though there may be a contrast between the brightly lit and dimly lit phases which, in the absence of darkness, would cause the dim phase to be treated as dark.