Synthesis and use of 6,6,6-trifluoro-L-fucose to block core-fucosylation in hybridoma cell lines.

Many monoclonal antibodies (mAbs) used in cancer immunotherapy mediate tumour cell lysis by recruiting natural killer (NK) cells; a phenomenon known as antibody-dependent cellular cytotoxicity (ADCC). Eliminating core-fucose from the N-glycans of a mAb enhances its capacity to induce ADCC. As such, inhibitors of fucosylation are highly desirable for the production of mAbs for research and therapeutic use. Herein, we describe a simple synthesis of 6,6,6-trifluoro-l-fucose (F3Fuc), a metabolic inhibitor of fucosylation, and demonstrate the utility of this molecule in the production of low-fucose mAbs from murine hybridoma cell lines.

Protein O-fucosylation in Plasmodium falciparum ensures efficient infection of mosquito and vertebrate hosts.

O-glycosylation of the Plasmodium sporozoite surface proteins CSP and TRAP was recently identified, but the role of this modification in the parasite life cycle and its relevance to vaccine design remain unclear. Here, we identify the Plasmodium protein O-fucosyltransferase (POFUT2) responsible for O-glycosylating CSP and TRAP. Genetic disruption of POFUT2 in Plasmodium falciparum results in ookinetes that are attenuated for colonizing the mosquito midgut, an essential step in malaria transmission. Some POFUT2-deficient parasites mature into salivary gland sporozoites although they are impaired for gliding motility, cell traversal, hepatocyte invasion, and production of exoerythrocytic forms in humanized chimeric liver mice. These defects can be attributed to destabilization and incorrect trafficking of proteins bearing thrombospondin repeats (TSRs). Therefore, POFUT2 plays a similar role in malaria parasites to that in metazoans: it ensures the trafficking of Plasmodium TSR proteins as part of a non-canonical glycosylation-dependent endoplasmic reticulum protein quality control mechanism.The role of O-glycosylation in the malaria life cycle is largely unknown. Here, the authors identify a Plasmodium protein O-fucosyltransferase and show that it is important for normal trafficking of a subset of surface proteins, particularly CSP and TRAP, and efficient infection of mosquito and vertebrate hosts.

Age-dependent differences in the attentional demands of obstacle negotiation.

BACKGROUND: Although the attention directed to gait does increase as an impending obstacle nears, it remains unclear whether the allocation of attentional resources differs between the precrossing phase of obstacle negotiation and the crossing phase of this task. This study compared the attentional demands associated with steady-state walking and the precrossing and crossing phases of an obstacle-negotiation task between young and older adults. METHODS: Fifteen younger and 15 older adults participated in this study. Participants were required to perform a verbal reaction time task during three events: (1) steady-state unobstructed gait, (2) precrossing (the final full stride prior to obstacle crossing), and (3) obstacle crossing. RESULTS: The attention directed during precrossing exceeds that of steady-state walking for both younger and older adults. Younger adults direct more attention to gait during precrossing than they do during crossing. However, precrossing is equally attentionally demanding as crossing for older adults, and both of these events require more attention than does steady-state gait in this age group. CONCLUSIONS: The task of obstacle negotiation, from precrossing through obstacle crossing, is attentionally demanding for elderly persons, and fall risk, due to a compromised availability of attentional resources, does occur prior to obstacle crossing in this age group.

Obstacle negotiation kinematics: age-dependent effects of postural threat.

This study investigated whether obstacle negotiation kinematics among younger (YA) and older adults (OA) are influenced by postural threat. Obstacle negotiation kinematics among YA and OA were examined under four conditions of postural threat. Seventeen older and 15 YA negotiated a fixed virtual obstacle while walking at a self-determined velocity along a 7.2 m walkway. Postural threat was manipulated by varying the width (0.60 vs. 0.15 m) and height (0.00 vs. 0.60 m) of the walkway. Increasing postural threat resulted in elevated levels of physiological arousal and altered crossing kinematics for all subjects. Specifically, crossing step length, lead limb velocity, trail limb velocity, and whole body center of mass (COM) velocity decreased and lead limb crossing height increased in the condition of greatest postural threat compared with the condition of least postural threat. Although both YA and OA altered obstacle negotiation kinematics under conditions of postural threat, the changes observed among older adults were considerably different from those of YA. In particular, OA demonstrated more marked reductions in the crossing velocity of the lead limb, trail limb, and COM than YA between the condition of least postural threat and the condition of greatest postural threat. The results of this study reveal that postural threat influences negotiation kinematics among YA and OA and illustrate that age-dependent differences exist for obstacle negotiation kinematics when postural threat increases. These findings may imply a beneficial effect of fear of falling on reducing fall probability among healthy OA.

The allocation of attention during locomotion is altered by anxiety.

We tested the hypotheses that: (1) anxiety regarding the possibility of falling alters the attentional demands of gait; and (2) this alteration in the attentional requirements of gait occurs independently of gait pattern accommodations. Sixteen younger and 15 older adults participated in this study. Subjects walked at a self-determined velocity along a 7.2-m walkway under four conditions of postural threat; the walking conditions varied depending on the width constraints of the walkway (60 cm vs 15 cm) and the height of the walking surface (0 cm vs 60 cm). Attentional demands of locomotion in each condition of testing were assessed using the dual-task methodology, in which participants verbally responded to an auditory cue as quickly as possible while continuing to walk. Our findings revealed that: (1) participants were successful in verbally responding to the auditory cue without modifying their gait pattern; (2) reaction times increased for all subjects when walking in the condition of greatest postural threat; (3) the attentional demands for locomotion varied with the phase of the gait cycle for younger adults but not for older adults; (4) the phase-dependent effect for younger adults disappeared in the condition of greatest postural threat, while reaction time scores for older adults systematically increased; and (5) increases in reaction time persisted despite significant changes in gait kinematics. Our findings confirm that anxiety increases the attentional demands for locomotion and provide further explanation for age-dependent increases in the attentional demands of gait. Furthermore, our findings confirm that fall-related anxiety predicates an increase in the allocation of attention to locomotor control that is independent of gait pattern adjustments.