Successful application of two-photon imaging with genetic tools in awake macaque monkeys will enable fundamental advances in our understanding of higher cognitive function at the level of molecular and neuronal circuits. Here we report techniques for long-term two-photon imaging in awake macaque monkeys. Using genetically encoded indicators including GCaMP5 and GCaMP6s delivered by AAV2/1 into the visual cortex, we demonstrate that high-quality two-photon imaging of large neuronal populations can be achieved and maintained in awake monkeys for months. Simultaneous intracellular recording and two-photon calcium imaging confirm that fluorescence activity is linearly proportional to neuronal spiking activity across a wide range of firing rates (10 Hz to 150 Hz). By providing two-photon imaging access to cortical neuronal populations at single-cell or single dendritic spine resolution in awake monkeys, the techniques reported can help bridge the use of modern genetic and molecular tools and the study of higher cognitive function.

Seed number per unit area is important in determining yield in soybean [Glycine max (L.) Merr.]. Field and greenhouse experiments were conducted to evaluate the role of flowers per plant and flower and fruit abscission in determining pod and seed numbers. Two greenhouse experiments were conducted with the McCall cultivar [Maturity Group (MG) 00] and four cultivars [McCall, Hardin (MG I), Harper (MG III), Essex (MG V)] were used in field experiments in 1989 and 1990. The field experiments used either 0.38‐m (McCall and Hardin) or 0.76‐m rows (Harper and Essex) with a constant seeding rate of 26 seeds per meter. Shade cloth (63% in the greenhouse and 30 and 63% in the field) was placed over plants from R1 to maturity. Additional treatments in the greenhouse included shade from R1 to R4, R4 to R5, R5 to R6, and R4 to R8. Plants were harvested at maturity and flower and pod abscission was determined by counting both the scars marking the point of attachment of flowers and pods, and mature pods. Shade reduced pod numbers and the reductions were due to both increased flower and pod abscission and fewer flowers per plant. Both the number of nodes on the main stem and flowers per node were affected by the shade. Shade during any part of the R1 to R6 period reduced pod numbers in the greenhouse. These data indicate that environmentally induced variation in pod numbers in soybean is a result of both changes in flower and fruit abscission and the number of flowers produced.

PD-L1 up-regulation in cancer contributes to immune evasion by tumor cells. Here, we show that Wnt ligand and activated EGFR induce the binding of the β-catenin/TCF/LEF complex to the CD274 gene promoter region to induce PD-L1 expression, in which AKT activation plays an important role. β-Catenin depletion, AKT inhibition, or PTEN expression reduces PD-L1 expression in tumor cells, enhances activation and tumor infiltration of CD8+ T cells, and reduces tumor growth, accompanied by prolonged mouse survival. Combined treatment with a clinically available AKT inhibitor and an anti–PD-1 antibody overcomes tumor immune evasion and greatly inhibits tumor growth. In addition, AKT-mediated β-catenin S552 phosphorylation and nuclear β-catenin are positively correlated with PD-L1 expression and inversely correlated with the tumor infiltration of CD8+ T cells in human glioblastoma specimens, highlighting the clinical significance of β-catenin activation in tumor immune evasion.