Analyses of thermoregulatory responses of feeder cattle exposed to simulated heat waves

Heat stress in feedlot cattle causes reduced performance, and in the most severe cases, death of the animals, thus causing the loss of millions of dollars in revenue to the cattle industry. A study was designed to evaluate dynamics of thermoregulation and feeding activities when feeder cattle were exposed to simulated heat waves, in comparison with repeated sinusoidal hot and thermoneutral environments. Nine beef steers were randomly assigned to an individual pen in one of three environmental chambers. Each chamber was subjected to each of three temperature regimes (Heatwave simulation from Rockport, Mo., 1995, Heatwave simulation from Columbia, Mo., 1999, and Controlled heat stress treatment of 32±7°C) for a period of 18 days, according to a Latin square treatment design, with a 10-day thermoneutral period (18±7°C) separating treatment periods. Respiration rate, core body temperature, heat production, feed intake, and feeding behavior were measured on each animal for the duration of the experiment. Differences were found in all treatments for all parameters except feeding behavior. It was shown that the two simulated heat waves elicited very different thermoregulatory responses. Based on these results the heat wave centered at Rockport, Mo. in 1995 was devastating because the animals were not acclimated to hot conditions, thus causing an acute response to heat stress. The responses of cattle to conditions at Columbia, Mo. showed some acclimation to heat prior to the peak stress days, and therefore a dampened response was seen. It appears the extreme conditions at Columbia, Mo., 1999 were made severe by environmental conditions not simulated during this study (low wind speed and intensive solar radiation). Overall, it was determined while a cyclic heat stress treatment is a representative model to test heat stress in cattle, further heat stress experiments should be conducted in an actual feedlot.Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. 9th Street Drive, West Palmetto, FL 34221, USA.     

Molluscicidal saponins of Phytolacca dodecanra: Oleanoglycotoxin-A

The structure of one of the major molluscicidal saponins of the fruit of Phytolacca dodecandra has been elucidated as 3-[2,4-di-O-(β-d-glucopyranosyl)-β-d-glucopyranosyll-olean-12-ene-28-oic acid. The combined use of 300 Mc. PMR, MS and GC-MS led to this structural assignment.     

The Drosophila STUbL protein Degringolade limits HES functions during embryogenesis

Degringolade (Dgrn) encodes a Drosophila SUMO-targeted ubiquitin ligase (STUbL) protein similar to that of mammalian RNF4. Dgrn facilitates the ubiquitylation of the HES protein Hairy, which disrupts the repressive activity of Hairy by inhibiting the recruitment of its cofactor Groucho. We show that Hey and all HES family members, except Her, interact with Dgrn and are substrates for its E3 ubiquitin ligase activity. Dgrn displays dynamic subcellular localization, accumulates in the nucleus at times when HES family members are active and limits Hey and HES family activity during sex determination, segmentation and neurogenesis. We show that Dgrn interacts with the Notch signaling pathway by it antagonizing the activity of E(spl)-C proteins. dgrn null mutants are female sterile, producing embryos that arrest development after two or three nuclear divisions. These mutant embryos exhibit fragmented or decondensed nuclei and accumulate higher levels of SUMO-conjugated proteins, suggesting a role for Dgrn in genome stability.     

Cell wound repair in Drosophila occurs through three distinct phases of membrane and cytoskeletal remodeling

When single cells or tissues are injured, the wound must be repaired quickly in order to prevent cell death, loss of tissue integrity, and invasion by microorganisms. We describe Drosophila as a genetically tractable model to dissect the mechanisms of single-cell wound repair. By analyzing the expression and the effects of perturbations of actin, myosin, microtubules, E-cadherin, and the plasma membrane, we define three distinct phases in the repair process-expansion, contraction, and closure-and identify specific components required during each phase. Specifically, plasma membrane mobilization and assembly of a contractile actomyosin ring are required for this process. In addition, E-cadherin accumulates at the wound edge, and wound expansion is excessive in E-cadherin mutants, suggesting a role for E-cadherin in anchoring the actomyosin ring to the plasma membrane. Our results show that single-cell wound repair requires specific spatial and temporal cytoskeleton responses with distinct components and mechanisms required at different stages of the process.     

A TCR targeting the HLA-A*0201-restricted epitope of MAGE-A3 recognizes multiple epitopes of the MAGE-A antigen superfamily in several types of cancer

Adoptive immunotherapy using TCR-engineered PBLs against melanocyte differentiation Ags mediates objective tumor regression but is associated with on-target toxicity. To avoid toxicity to normal tissues, we targeted cancer testis Ag (CTA) MAGE-A3, which is widely expressed in a range of epithelial malignancies but is not expressed in most normal tissues. To generate high-avidity TCRs against MAGE-A3, we employed a transgenic mouse model that expresses the human HLA-A*0201 molecule. Mice were immunized with two HLA-A*0201-restricted peptides of MAGE-A3: 112-120 (KVAELVHFL) or MAGE-A3: 271-279 (FLWGPRALV), and T cell clones were generated. MAGE-A3-specific TCR α- and β-chains were isolated and cloned into a retroviral vector. Expression of both TCRs in human PBLs demonstrated Ag-specific reactivity against a range of melanoma and nonmelanoma tumor cells. The TCR against MAGE-A3: 112-120 was selected for further development based on superior reactivity against tumor target cells. Interestingly, peptide epitopes from MAGE-A3 and MAGE-A12 (and to a lesser extent, peptides from MAGE-A2 and MAGE-A6) were recognized by PBLs engineered to express this TCR. To further improve TCR function, single amino acid variants of the CDR3 α-chain were generated. Substitution of alanine to threonine at position 118 of the α-chain in the CDR3 region of the TCR improved its functional avidity in CD4 and CD8 cells. On the basis of these results, a clinical trial is planned in which patients bearing a variety of tumor histologies will receive autologous PBLs that have been transduced with this optimized anti-MAGE-A3 TCR.     

The KASH domain protein MSP-300 plays an essential role in nuclear anchoring during Drosophila oogenesis

During late stages of Drosophila oogenesis, the cytoplasm of nurse cells in the egg chamber is rapidly transferred ("dumped") to oocytes, while the nurse cell nuclei are anchored by a mechanism that involves the actin cytoskeleton. The factors that mediate this interaction between nuclei and actin cytoskeleton are unknown. MSP-300 is the likely Drosophila ortholog of the mammalian Syne-1 and -2 and C. elegans ANC-1 proteins, contained both actin-binding and nuclear envelope localization domains. By using an antibody against C-terminus of MSP-300, we find that MSP-300 is distributed throughout the cytoplasm and accumulates at the nuclear envelope of nurse cells and the oocyte. A GFP fusion protein containing the C-terminal region of MSP-300 is also sufficient to localize protein on the nuclear envelope in oocytes. To eliminate the maternal gene activity during oogenesis, we generated homozygous germ-line clones of a loss-of-function mutation in msp-300 in otherwise heterozygous mothers. In the mutant egg chambers that develop from such clones, cytoplasmic dumping of nurse cells is severely disturbed. The nuclei of nurse cells and the oocyte are mislocalized and the usually well-organized actin structures are severely disrupted. These results indicate that maternal MSP-300 plays an important role in actin-dependent nuclear anchorage during cytoplasmic transport.