Sequence-specific 2'-O-methoxyethyl antisense oligonucleotides activate human platelets through glycoprotein VI, triggering formation of platelet-leukocyte aggregates.

Antisense oligonucleotides (ASO) are DNA-based, disease-modifying drugs. Clinical trials with 2'-O-methoxyethyl (2'MOE) ASO have shown dose- and sequence-specific lowering of platelet counts according to two phenotypes. Phenotype 1 is a moderate (but not clinically severe) drop in platelet count. Phenotype 2 is rare, severe thrombocytopenia. This article focuses on the underlying cause of the more common phenotype 1, investigating the effects of ASO on platelet production and platelet function. Five phosphorothioate ASO were studied: three 2'MOE sequences; 487660 (no effects on platelet count), 104838 (associated with phenotype 1), and 501861 (effects unknown) and two CpG sequences; 120704 and ODN 2395 (known to activate platelets). Human cord bloodderived megakaryocytes were treated with these ASO to study their effects on proplatelet production. Platelet activation (determined by surface Pselectin) and platelet-leukocyte aggregates were analyzed in ASO-treated blood from healthy human volunteers. None of the ASO inhibited proplatelet production by human megakaryocytes. All the ASO were shown to bind to the platelet receptor glycoprotein VI (KD ~0.2-1.5 mM). CpG ASO had the highest affinity to glycoprotein VI, the most potent platelet-activating effects and led to the greatest formation of platelet-leukocyte aggregates. 2'MOE ASO 487660 had no detectable platelet effects, while 2'MOE ASOs 104838 and 501861 triggered moderate platelet activation and SYKdependent formation of platelet-leukocyte aggregates. Donors with higher platelet glycoprotein VI levels had greater ASO-induced platelet activation. Sequence-dependent ASO-induced platelet activation and platelet-leukocyte aggregates may explain phenotype 1 (moderate drops in platelet count). Platelet glycoprotein VI levels could be useful as a screening tool to identify patients at higher risk of ASO-induced platelet side effects.

Identification of abscission checkpoint bodies as structures that regulate ESCRT factors to control abscission timing.

The abscission checkpoint regulates the ESCRT membrane fission machinery and thereby delays cytokinetic abscission to protect genomic integrity in response to residual mitotic errors. The checkpoint is maintained by Aurora B kinase, which phosphorylates multiple targets, including CHMP4C, a regulatory ESCRT-III subunit necessary for this checkpoint. We now report the discovery that cytoplasmic abscission checkpoint bodies (ACBs) containing phospho-Aurora B and tri-phospho-CHMP4C develop during an active checkpoint. ACBs are derived from mitotic interchromatin granules, transient mitotic structures whose components are housed in splicing-related nuclear speckles during interphase. ACB formation requires CHMP4C, and the ESCRT factor ALIX also contributes. ACB formation is conserved across cell types and under multiple circumstances that activate the checkpoint. Finally, ACBs retain a population of ALIX, and their presence correlates with delayed abscission and delayed recruitment of ALIX to the midbody where it would normally promote abscission. Thus, a cytoplasmic mechanism helps regulate midbody machinery to delay abscission.

When a cell divides, it must first carefully duplicate its genetic information and package these copies into compartments housed in the two new cells. Errors in this process lead to genetic mistakes that trigger cancer or other harmful biological events. Quality control checks exist to catch errors before it is too late. This includes a final 'abscission' checkpoint right before the end of division, when the two new cells are still connected by a thin membrane bridge. If cells fail to pass this 'no cut' checkpoint, they delay severing their connection until the mistake is fixed. A group of proteins called ESCRTs is responsible for splitting the two cells apart if nothing is amiss. The abscission checkpoint blocks this process by altering certain proteins in the ESCRT complex, but exactly how this works is not yet clear. To find out more, Williams et al. imaged ESCRT factors in a new experimental system in which the abscission checkpoint is active in many cells. This showed that, in this context, certain ESCRT components were rerouted from the thread of membrane between the daughter cells to previously unknown structures, which Williams et al. named abscission checkpoint bodies. These entities also sequestered other factors that participate in the abscission checkpoint and factors that contribute to gene expression. These results are key to better understand how cells regulate their division; in particular, they provide a new framework to explore when this process goes wrong and contributes to cancer.

Analysis of ants' rescue behavior reveals heritable specialization for first responders.

In colonies of Cataglyphis cursor ants, a single queen mates with multiple males, creating the foundation for heritable behavioral specializations. A novel and unique candidate for such specializations is rescue behavior, a precisely delivered form of altruism in which workers attempt to release trapped nestmates and which relies on short-term memory of previous actions to increase its efficiency. Consistent with task specialization, not all individuals participate; instead, some individuals move away from the victim, which gives rescuers unrestricted access. Using a bioassay to identify rescuers and non-rescuers, coupled with paternity assignment via polymorphic microsatellite markers, we not only show that rescue behavior is heritable, with 34% of the variation explained by paternity, but also establish that rescue, heretofore overlooked in analyses of division of labor, is a true specialization, an ant version of first responders. Moreover, this specialization emerges as early as 5 days of age, and the frequency of rescuers remains constant across ants' age ranges. The extremely broad range of these ants' heritable polyethism provides further support for the critical role of polyandry in increasing the efficiency of colony structure and, in turn, reproductive success.

Modulation of megakaryopoiesis and platelet production during inflammation.

Megakaryocytes (MKs) are widely known as the progenitor cells of platelets. These large, polyploid cells are a derivative of the hematopoietic stem cell (HSC), and reside in the bone marrow, lining blood vessel walls where they release their platelet progeny into circulation. Although little is known about how MKs differ under various environmental stressors, both chronic and acute inflammation alter the differentiation and molecular content of MKs. Furthermore, evidence suggests that the release of inflammatory cytokines may induce MK rupture and rapid release of platelets as a mechanism to quickly replenish diminished platelet counts in response to inflammation. Similarities between MKs and their close relatives, white blood cells, have introduced the notion that MKs may play a role in combating infection by engulfing and presenting antigens, and passing this information to circulating platelets. In addition, MKs exposed to varying bone marrow environments produce different platelets which enter circulation primed to respond to and combat inflammation, infection, or injury. This review focuses on how inflammation alters MK production, maturation, and platelet production. In addition, it introduces the idea that inflammation reprograms MKs to create different, more pathogenic platelets and leads them to take on different roles as responders to deleterious conditions. In the future, studies determining how platelets are altered in disease states may lead to novel MK- and platelet-based therapeutic targets to mitigate inflammation-related morbidity and mortality.

Exposure to a nicotinoid pesticide reduces defensive behaviors in a non-target organism, the rusty crayfish Orconectes rusticus.

Imidacloprid is the most widely used of the nicotinoid insecticides, the fastest growing class of pesticides on the global market. Although less toxic to mammals and birds compared to organophosphates, nicotinoids have the potential to impact non-target invertebrates, especially through sublehal effects on behavior, physiology, reproduction, and development. We investigated the impact of sublethal doses of imidacloprid on the defensive responses of rusty crayfish Orconectes rusticus exposed to 0, 1, 10, and 100 µg•L-1 of imidacloprid for 10 days (n = 7 crayfish per treatment). Defensive behaviors were examined with the rod test, in which a glass rod was jabbed into the crayfish's container at a 90 degree angle from the bottom and about 0.5 cm directly in front of the crayfish. Crayfish responded to the rod aggressively with claw raising and pinching, neutrally (no response), or by backing or tail-flipping away. The frequency of neutral responses more than doubled after four days in the high (100 µg•L-1) group and after eight days in the low (1 µg•L-1) exposure group. Furthermore, most crayfish in the 100 µg•L-1 treatment were not able to right themselves within 30 s when placed on their backs. Several studies have reported concentrations of imidacloprid contamination in freshwater ecosystems that exceed this study's lowest exposure scenario, 1 µg•L-1. We therefore conclude that imidacloprid contamination reduces the defensive behaviors of crayfish, impairing their ability to survive in habitats where they play important ecological roles.

Pituitary macroadenomas with oculomotor cistern extension and tracking: implications for surgical management.

OBJECT Oculomotor cistern extension of pituitary adenomas is an overlooked feature within the literature. In this study, 7 cases of pituitary macroadenoma with oculomotor cistern extension and tracking are highlighted, and the implications of surgical and medical management are discussed. METHODS The records of patients diagnosed with pituitary macroadenomas who underwent resection and in whom preoperative pituitary protocol MRI scans were available for review were retrospectively reviewed. The patient and tumor characteristics were reviewed along with the operative outcomes and complications. RESULTS Seven patients (4.1%) with oculomotor cistern extension and tracking were identified in a cohort of 170 patients with pituitary macroadenoma. The most common presenting symptoms were visual deficit (6 patients; 86%), apoplexy (3 patients; 43%), and oculomotor nerve palsy (3 patients; 43%). Lone oculomotor nerve palsy was seen in 2 patients without apoplexy and 1 patient with an apoplectic event. Gross-total resection was achieved via a microscopic endonasal transsphenoidal approach with or without endoscopic aid to the sella in 14%, near-total resection in 29%, and subtotal resection in 57% of patients in the data set. CONCLUSIONS Pituitary adenoma extension along the oculomotor cistern is uncommon; however, preoperatively recognizing such extension should play an important role in the surgeon's operative considerations and postoperative clinical management because this extension can limit gross-total resection using the transsphenoidal approach alone.

Peritumoral cysts associated with pituitary macroadenoma.

OBJECT: Peritumoral cysts are benign nonneoplastic cysts that are found adjacent to extraaxial brain tumors such as meningiomas, schwannomas, craniopharyngiomas, and esthesioneuroblastomas. Peritumoral cysts associated with pituitary macroadenomas have not been previously described in the literature. The authors report 6 cases of giant macroadenoma-associated peritumoral cysts and delineate their imaging spectrum. METHODS: The authors retrospectively reviewed the records of 179 patients diagnosed with pituitary macroadenomas who underwent tumor resection at their institution and had preoperative MRI scans available for review. The patients were evaluated for the presence of associated peritumoral cysts. Clinical presentation, histopathology, follow-up time, tumor and peritumoral cyst dimensions were recorded. Signal intensity on T1-weighted, T2-weighted, diffusion-weighted, and FLAIR sequences, as well as pre- and postcontrast appearance, were determined. RESULTS: Six patients (3.4%) with associated peritumoral cysts were identified in our cohort of 179 patients with pituitary macroadenoma. Twelve patients in the cohort had giant macroadenomas (≥ 4.0 cm), and 50% of these tumors had associated peritumoral cysts with significant extrasellar extension of the macroadenoma. Only tumors with craniocaudal, transverse, and anteroposterior diameters of 3.6 × 3.4 × 4.2 cm to 7.0 × 7.4 × 6.8 cm (mean 5.3 × 5.1 × 5.6 cm), respectively, had associated peritumoral cysts. The growth pattern in all tumors was suprasellar, with predominant anterior and lateral extension. Cysts showed T1-weighted, T2-weighted, and FLAIR hyperintensity in 67%, 67%, and 60% of patients, respectively. There was no contrast enhancement of the cyst wall or fluid contents in any patient. Postoperatively, cysts had completely resolved (4 of 5) or significantly decreased in size (1 of 5). One patient was lost to follow-up. CONCLUSIONS: Macroadenoma-associated peritumoral cysts are rare, benign, and likely nonneoplastic fluid collections that do not represent neoplasm. These cysts display a predictable pattern of hyperintensity on T1-weighted, T2-weighted, and FLAIR sequences and do not enhance. They most likely represent proteinaceous CSF in a sulcus or cistern that becomes trapped (encysted) by anterolateral extension of unusually large macroadenomas. Peritumoral cysts may facilitate resection of the associated macroadenoma by providing a cleavage plane.