Anti-Leukemic Properties of Aplysinopsin Derivative EE-84 Alone and Combined to BH3 Mimetic A-1210477

Aplysinopsins are a class of marine indole alkaloids that exhibit a wide range of biological activities. Although both the indole and N-benzyl moieties of aplysinopsins are known to possess antiproliferative activity against cancer cells, their mechanism of action remains unclear. Through in vitro and in vivo proliferation and viability screening of newly synthesized aplysinopsin analogs on myelogenous leukemia cell lines and zebrafish toxicity tests, as well as analysis of differential toxicity in noncancerous RPMI 1788 cells and PBMCs, we identified EE-84 as a promising novel drug candidate against chronic myeloid leukemia. This indole derivative demonstrated drug-likeness in agreement with Lipinski’s rule of five. Furthermore, EE-84 induced a senescent-like phenotype in K562 cells in line with its cytostatic effect. EE-84-treated K562 cells underwent morphological changes in line with mitochondrial dysfunction concomitant with autophagy and ER stress induction. Finally, we demonstrated the synergistic cytotoxic effect of EE-84 with a BH3 mimetic, the Mcl-1 inhibitor A-1210477, against imatinib-sensitive and resistant K562 cells, highlighting the inhibition of antiapoptotic Bcl-2 proteins as a promising novel senolytic approach against chronic myeloid leukemia.     

A comparison of the effects of the nematodes Radopholus similis and Pratylenchus goodeyi on growth,root health and yield of an East African highland cooking banana (Musa AAA-group)

We combined two protein-marking enzyme-linked immunosorbent assays (ELISA) with a predator gut content ELISA to monitor the movement and feeding activity of commercially-purchased Hippodamia convergens Guèrin-Mèneville (Coleoptera: Coccinellidae) under realistic field conditions during two field seasons in central Arizona. The protein-marking ELISAs were used to differentiate released H. convergens from the native beetles. Commercially purchased beetles marked with rabbit immunoglobulin G (IgG) were released into cotton fields and chicken IgG marked beetles were released into adjacent cantaloupe fields. Results showed that the total native beetle abundance in each crop was about the same size. The recovery rates after 15 days for the released beetles were less than 1.0% over all the releases, indicating that they dispersed readily from the release site. Of the recaptured beetles containing rabbit IgG (cotton), 82.2% were recovered in cotton and 11.8% moved to cantaloupe. Of those containing chicken IgG (cantaloupe), 66.5% were recovered in cantaloupe and 33.5% moved to cotton. A predator gut content ELISA was used to determine if there were differences in the frequency of predation of released versus indigenous H. convergens on the silverleaf whitefly, Bemisia argentifolii Bellows &; Perring (Homoptera: Aleyrodidae). The proportion of beetles containing whitefly antigens was always higher for the released beetles than for their native counterparts. Our results demonstrate an approach to combine protein marking and predator gut content ELISAs that allows the simultaneous comparison of feeding and intercrop movement of native and commercially-obtained biological control agents.     

The use of electrical stunning followed by electro-immobilization for the humane slaughter of cattle

Sir;- In New Zealand, calves are often electrically stunned before slaughter. However, electrical stunning is not usually used for adult cattle, primarily because the large size of the adult animal makes restraint more difficult, resulting in less reproducible stunning, with potential danger to personnel from animal movement. In addition, the slaughter of cattle may be inhumane if the stunning process does not ensure immediate and permanent insensibility (Newhook and Blackmore 1982b). The use and humaneness of electrical stunning of sheep and cattle have been the subject of our recent research, in which we have addressed and resolved many of these problems. We wish to relate our observations in the context of head-only electrical stunning of cattle followed by electro-immobilization to maintain insensibility and ensure carcass stillness.     

Factors Affecting the Reproductive Efficiency of Artificial Insemination Programmes in a Seasonal Breeding Pasture-based Dairy System with the Aid of Milk Progesterone

To determine the factors affecting reproductive efficiency in a seasonal breeding artificial insemination (AI) programme in dairy herds in Uruguay, a field trial was conducted on three dairy farms. Lactating cows (n=328) intended to be bred at the following breeding period were selected. The trial started 1 week before the beginning of the breeding period and lasted 80 days or until first service. Milk samples were obtained biweekly for 12 weeks, starting 1 week prior to the breeding season to determine progesterone concentrations. Data indicated that 12.5% of the cows were anoestrus at the beginning of the season remaining in this condition throughout the trial. Most of the anoestrous animals were first-calf heifers. A varying number of cycling cows (8.5%) at each farm were never reported in heat. Oestrous detection rate for cows cycling, as determined by progesterone patterns, was evaluated in three periods of 21 days and resulted in an overall efficiency of 46.9%. Main factors affecting oestrous detection were farm, with an effect of parity (67.8%, mature cows and 33.2%, first-calf heifers) and no effect of days postpartum. Mean interval from the beginning of the breeding season to first breeding was 27.4 days, which was also affected by farm. Neither parity nor days postpartum affected this parameter.     

Effects of Glucose Concentration on in vitro Fertilization in BALB/c Mice

BALB/c mice are widely used in genetic, tumour and immunological studies. However, the mice demonstrate a lower reproduction rate, low fertility and small litters, because of their highly genetic homozygoisty. Based on in vitro fertilization (IVF), a routine technique for biomedical studies, it is worth to evaluate the effects to BALB/c mice on IVF efficiency. In order to test the genetic factor affecting the IVF efficiency of BALB/c, four reciprocal IVF tests of BALB/cByJ and FVB/NCrl mice were performed. The results showed that the average fertility of IVF sponsored by FVB/NCrl spermatozoa was 69.6%, but only 12.1% was obtained from BALB/cByJ strain. Effect of glucose contained in the culture medium to the IVF efficiency of BALB/cByJ was also evaluated. The results showed that the fertility of BALB/cByJ spermatozoa incubated with 0, 2.7, 5.5, 11.1 and 22.2 mm of glucose in the TYH medium were 6.8, 9.9, 13.9, 32.7 and 22.2%, respectively. It is showed that IVF efficiency of BALB/cByJ spermatozoa could be improved depending on the concentration of glucose in the IVF medium. According to the results, it is beleived that lower IVF of BALB/cByJ mice might be due to the genetic defect in spermatozoa and increasing glucose in the IVF medium which significantly affect the IVF efficiency of BALB/cByl via activating the spermatozoa.     

Stifle joint osteoarthritis at the time of diagnosis of cranial cruciate ligament injury is higher in Boxers and in dogs weighing more than 35 kilograms

Osteoarthritis is a ubiquitous disease in dogs. The purpose of this retrospective study was to characterize the severity and distribution of osteoarthritis (OA) within the joint and to identify differences among dog breeds in the severity of OA in the cranial cruciate ligament (CCL)‐deficient stifle joint. Radiographs of 240 stifles from 51 Boxers, 66 German Shepherds, 100 Labrador Retrievers, and 23 Siberian Huskies with confirmed CCL rupture were included. Radiographs of the stifle joint were evaluated and OA severity was graded at 33 sites within and around the joint, and patella alta was graded as present or absent for a potential total stifle OA score of 100. Osteophyte size was correlated to OA severity score. Total OA scores were calculated and compared within and between breeds globally as well as at each joint site. Dogs weighing >35 kg had a higher total OA score than those weighing <35 kg. Osteoarthritis scores were highest at the apical patella, proximolateral tibia, and sesamoid bones, corresponding to the proximal, lateral, and caudal aspects of the joint, respectively. No statistically significant differences were found among the mean OA scores of various stifle joint regions. Boxer dogs had a higher total OA score than other breeds. We concluded that dogs have a consistent distribution pattern of OA within the stifle joint after CCL injury. Radiographic OA is more severe in the proximal, lateral, and caudal aspects of the joint. Boxers had more severe OA than the other breeds evaluated in the study.     

Prospection: experiencing the future

All animals can predict the hedonic consequences of events they've experienced before. But humans can predict the hedonic consequences of events they've never experienced by simulating those events in their minds. Scientists are beginning to understand how the brain simulates future events, how it uses those simulations to predict an event's hedonic consequences, and why these predictions so often go awry.     

Maternal undernutrition during early to mid-gestation in the ewe results in altered growth, adiposity, and glucose tolerance in male offspring

This study utilized maternal undernutrition from early to midgestation in the ewe to determine the impact(s) of intrauterine growth restriction on postpartum growth of male offspring and the potential mechanisms involved. Multiparous ewes were fed 50% (nutrient-restricted) or 100% (control-fed) of their nutrient requirements (NRC, 1985) between d 28 and 78 of gestation, and then all ewes were fed 100% of the NRC requirements from d 79 through lambing. Male lambs born to nutrient-restricted (n = 9) and control-fed (n = 9) ewes exhibited similar BW (5.8 vs. 6.0 +/- 0.3 kg) and crown-rump lengths (53.8 vs. 55.4 +/- 1.0 cm) at birth. At 63 and 250 d of postnatal age, wether lambs were subjected to a glucose tolerance test, in which a bolus of glucose was administered i.v. to evaluate changes in glucose and insulin concentrations. After i.v. glucose administration at 63 d of age, lambs from nutrient-restricted ewes exhibited a greater area under the curve for glucose (AUCg; 6,281 vs. 5,242 +/- 429; P < 0.05) and insulin (AUCi; 21.0 vs. 8.6 +/- 1.9; P < 0.001) than lambs from control-fed ewes. After glucose administration at 250 d of age, lambs from nutrient-restricted ewes had greater AUCg (7,147 vs. 5,823 +/- 361; P < 0.01) but a lower AUCi (6.4 vs. 10.2 +/- 1.9; P = 0.05) than lambs from control-fed ewes. Lambs from nutrient-restricted ewes were heavier (26.6 vs. 21.8 +/- 2.3 kg; P < 0.05) and had more backfat (0.30 vs. 0.21 +/- 0.03 cm, P < 0.05) by 4 mo of age than the lambs from control-fed ewes. At slaughter at 280 d of age, lambs from nutrient-restricted ewes remained heavier than lambs from control-fed ewes, had greater (P < 0.05) amounts of kidney and pelvic-area adipose tissue, and tended (P < 0.10) to have reduced LM and semitendinosus muscle weights as a percentage of HCW. These data demonstrate that a bout of maternal undernutrition during early to midgestation in sheep increased BW and fat deposition during adolescence and dysregulated glucose uptake in the absence of any change in birth weight.