A Photodynamic and Photochemotherapeutic Platinum-Iridium Charge-Transfer Conjugate for Anticancer Therapy.

The novel hetero-dinuclear complex trans,trans,trans-[PtIV(py)2(N3)2(OH)(µ-OOCCH2CH2CONHCH2-bpyMe)IrIII(ppy)2]Cl (Pt-Ir), exhibits charge transfer between the acceptor photochemotherapeutic Pt(IV) (Pt-OH) and donor photodynamic Ir(III) (Ir-NH2) fragments. It is stable in the dark, but undergoes photodecomposition more rapidly than the Pt(IV) parent complex (Pt-OH) to generate Pt(II) species, an azidyl radical and 1O2. The Ir(III)* excited state, formed after irradiation, can oxidise NADH to NAD⋅ radicals and NAD+. Pt-Ir is highly photocytotoxic towards cancer cells with a high photocytotoxicity index upon irradiation with blue light (465 nm, 4.8 mW/cm2), even with short light-exposure times (10-60 min). In contrast, the mononuclear Pt-OH and Ir-NH2 subunits and their simple mixture are much less potent. Cellular Pt accumulation was higher for Pt-Ir compared to Pt-OH. Irradiation of Pt-Ir in cancer cells damages nuclei and releases chromosomes. Synchrotron-XRF revealed ca. 4× higher levels of intracellular platinum compared to iridium in Pt-Ir treated cells under dark conditions. Luminescent Pt-Ir distributes over the whole cell and generates ROS and 1O2 within 1 h of irradiation. Iridium localises strongly in small compartments, suggestive of complex cleavage and excretion via recycling vesicles (e.g. lysosomes). The combination of PDT and PACT motifs in one molecule, provides Pt-Ir with a novel strategy for multimodal phototherapy.

Differential phase contrast for quantitative imaging and spectro-microscopy at a nanoprobe beamline.

The interaction of a focused X-ray beam with a sample in a scanning probe experiment can provide a variety of information about the interaction volume. In many scanning probe experiments X-ray fluorescence (XRF) is supplemented with measurements of the transmitted or scattered intensity using a pixelated detector. The automated extraction of different signals from an area pixelated detector is described, in particular the methodology for extracting differential phase contrast (DPC) is demonstrated and different processing methods are compared across a range of samples. The phase shift of the transmitted X-ray beam by the sample, extracted from DPC, is also compared with ptychography measurements to provide a qualitative and quantitative comparison. While ptychography produces a superior image, DPC can offer a simple, flexible method for phase contrast imaging which can provide fast results and feedback during an experiment; furthermore, for many science problems, such as registration of XRF in a lighter matrix, DPC can provide sufficient information to meet the experimental aims. As the DPC technique is a quantitative measurement, it can be expanded to spectroscopic studies and a demonstration of DPC for spectro-microscopy measurements is presented. Where ptychography can separate the absorption and phase shifts by the sample, quantitative interpretation of a DPC image or spectro-microscopy signal can only be performed directly when absorption is negligible or where the absorption contribution is known and the contributions can be fitted.

Does extended experience with other-race nannies predict racial bias in the preschool years?

Most existing studies on racial bias reduction have used short-term interracial interaction interventions with fleeting effects. The current natural experiment examined whether daily interactions with other-race nannies relate to reduced racial bias in the preschool years. We capitalized on a unique child-rearing situation in Singapore whereby children are often cared for by other-race nannies since infancy. Singaporean Chinese 3- to 6-year-olds (N = 100) completed explicit and implicit racial bias measures assessing their preferential bias favoring own-race adults over adults of their nannies' race. Differential findings were obtained for children's explicit and implicit racial bias. Extensiveness, but not mere presence, of other-race nanny experience was associated with lower levels of explicit racial bias in children. In contrast, neither presence nor extensiveness of other-race nanny experience was associated with children's implicit racial bias. Together, these findings suggest that long-term and extensive contact with an other-race caregiver could have subtle mitigating effects on children's explicit, but not implicit, racial bias.

Effects of face masks on language comprehension in bilingual children.

Due to the COVID-19 pandemic, many children receive language input through face coverings. The impact of face coverings for children's abilities to understand language remains unclear. Past research with monolingual children suggests that hearing words through surgical masks does not disrupt word recognition, but hearing words through transparent face shields proves more challenging. In this study, we investigated effects of different face coverings (surgical masks and transparent face shields) on language comprehension in bilingual children. Three-year-old English-Mandarin bilingual children (N = 28) heard familiar words in both English and Mandarin spoken through transparent face shields, surgical masks, and without masks. When tested in English, children recognized words presented without a mask and through a surgical mask, but did not recognize words presented with transparent face shields, replicating past findings with monolingual children. In contrast, when tested in Mandarin, children recognized words presented without a mask, through a surgical mask, and through a transparent face shield. Results are discussed in terms of specific properties of English and Mandarin that may elicit different effects for transparent face shields. Overall, the present findings suggest that face coverings, and in particular, surgical masks do not disrupt spoken word recognition in young bilingual children.

A cell design for correlative hard X-ray nanoprobe and electron microscopy studies of catalysts under in situ conditions.

To improve the understanding of catalysts, and ultimately the ability to design better materials, it is crucial to study them during their catalytic active states. Using in situ or operando conditions allows insights into structure-property relationships, which might not be observable by ex situ characterization. Spatially resolved X-ray fluorescence, X-ray diffraction and X-ray absorption near-edge spectroscopy are powerful tools to determine structural and electronic properties, and the spatial resolutions now achievable at hard X-ray nanoprobe beamlines make them an ideal complement to high-resolution transmission electron microscopy studies in a multi-length-scale analysis approach. The development of a system to enable the use of a commercially available gas-cell chip assembly within an X-ray nanoprobe beamline is reported here. The novel in situ capability is demonstrated by an investigation of the redox behaviour of supported Pt nanoparticles on ceria under typical lean and rich diesel-exhaust conditions; however, the system has broader application to a wide range of solid-gas reactions. In addition the setup allows complimentary in situ transmission electron microscopy and X-ray nanoprobe studies under identical conditions, with the major advantage compared with other systems that the exact same cell can be used and easily transferred between instruments. This offers the exciting possibility of studying the same particles under identical conditions (gas flow, pressure, temperature) using multiple techniques.

Undersampling raster scans in spectromicroscopy for a reduced dose and faster measurements.

Combinations of spectroscopic analysis and microscopic techniques are used across many disciplines of scientific research, including material science, chemistry and biology. X-ray spectromicroscopy, in particular, is a powerful tool used for studying chemical state distributions at the micro and nano scales. With the beam fixed, a specimen is typically rastered through the probe with continuous motion and a range of multimodal data is collected at fixed time intervals. The application of this technique is limited in some areas due to: long scanning times to collect the data, either because of the area/volume under study or the compositional properties of the specimen; and material degradation due to the dose absorbed during the measurement. In this work, we propose a novel approach for reducing the dose and scanning times by undersampling the raster data. This is achieved by skipping rows within scans and reconstructing the x-ray spectromicroscopic measurements using low-rank matrix completion. The new method is robust and allows for 5 to 6-fold reduction in sampling. Experimental results obtained on real data are illustrated.

Effects of interracial experience on the race preferences of infants.

Over their first year of life, infants express visual preferences for own- versus other-race faces. This developmental transition has primarily been investigated in monoracial societies where infants have limited personal or societal contact with other races. We investigated whether previously reported visual preferences for race generalize to a multiracial society (i.e., Singapore). In addition, we investigated effects of caregiver race on visual preferences for race. In Experiment 1, race preferences were measured at 3, 6, and 9 months of age for own-race (Chinese) versus other-race (Indian) faces in infants with no regular interaction with Indian-race individuals. Singaporean infants displayed a significant visual preference for Indian-race faces at each age group. Furthermore, infants raised with other-race caregivers demonstrated an age-related increase in other-race visual preferences. The visual preferences of infants for other-race faces were predicted by the extent of other-race contact. In Experiment 2, we confirmed that an other-race visual preference was not exclusive to Indian faces in a sample of 6-month-old Singaporean Chinese infants who demonstrated a similar other-race visual preference for Caucasian faces over Chinese faces. Findings are discussed in terms of the influence of other-race contact on visual preferences for race in infants.

Single-Cell Chemistry of Photoactivatable Platinum Anticancer Complexes.

The Pt(IV) prodrug trans, trans, trans-[Pt(pyridine)2(N3)2(OH)2] (Pt1) and its coumarin derivative trans, trans, trans-[Pt(pyridine)2(N3)2(OH)(coumarin-3-carboxylate)] (Pt2) are promising agents for photoactivated chemotherapy. These complexes are inert in the dark but release Pt(II) species and radicals upon visible light irradiation, resulting in photocytotoxicity toward cancer cells. Here, we have used synchrotron techniques to investigate the in-cell behavior of these prodrugs and visualize, for the first time, changes in cellular morphology and Pt localization upon treatment with and without light irradiation. We show that photoactivation of Pt2 induces remarkable cellular damage with extreme alterations to multiple cellular components, including formation of vacuoles, while also significantly increasing the cellular accumulation of Pt species compared to dark conditions. X-ray absorption near-edge structure (XANES) measurements in cells treated with Pt2 indicate only partial reduction of the prodrug upon irradiation, highlighting that phototoxicity in cancer cells may involve not only Pt(II) photoproducts but also photoexcited Pt(IV) species.

Beam and sample movement compensation for robust spectro-microscopy measurements on a hard X-ray nanoprobe.

Static and in situ nanoscale spectro-microscopy is now routinely performed on the Hard X-ray Nanoprobe beamline at Diamond and the solutions implemented to provide robust energy scanning and experimental operation are described. A software-based scheme for active feedback stabilization of X-ray beam position and monochromatic beam flux across the operating energy range of the beamline is reported, consisting of two linked feedback loops using extremum seeking and position control. Multimodal registration methods have been implemented for active compensation of drift during an experiment to compensate for sample movement during in situ experiments or from beam-induced effects.

The Hard X-ray Nanoprobe beamline at Diamond Light Source.

The Hard X-ray Nanoprobe beamline, I14, at Diamond Light Source is a new facility for nanoscale microscopy. The beamline was designed with an emphasis on multi-modal analysis, providing elemental mapping, speciation mapping by XANES, structural phase mapping using nano-XRD and imaging through differential phase contrast and ptychography. The 185 m-long beamline operates over a 5 keV to 23 keV energy range providing a ≤50 nm beam size for routine user experiments and a flexible scanning system allowing fast acquisition. The beamline achieves robust and stable operation by imaging the source in the vertical direction and implementing horizontally deflecting primary optics and an overfilled secondary source in the horizontal direction. This paper describes the design considerations, optical layout, aspects of the hardware engineering and scanning system in operation as well as some examples illustrating the beamline performance.

Infants recognize words spoken through opaque masks but not through clear masks.

COVID-19 has modified numerous aspects of children's social environments. Many children are now spoken to through a mask. There is little empirical evidence attesting to the effects of masked language input on language processing. In addition, not much is known about the effects of clear masks (i.e., transparent face shields) versus opaque masks on language comprehension in children. In the current study, 2-year-old infants were tested on their ability to recognize familiar spoken words in three conditions: words presented with no mask, words presented through a clear mask, and words presented through an opaque mask. Infants were able to recognize familiar words presented without a mask and when hearing words through opaque masks, but not when hearing words through clear masks. Findings suggest that the ability of infants to recover spoken language input through masks varies depending on the surface properties of the mask.

When novelty prevails on familiarity: Visual biases for child versus infant faces in 3.5- to 12-month-olds.

The current study examined the influence of everyday perceptual experience with infant and child faces on the shaping of visual biases for faces in 3.5-, 6-, 9-, and 12-month-old infants. In Experiment 1, infants were presented with pairs of photographs of unfamiliar child and infant faces. Four groups with differential experience with infant and child faces were composed from parents' reports of daily exposure with infants and children (no experience, infant face experience, child face experience, and both infant and child face experience) to assess influence of experience on face preferences. Results showed that infants from all age groups displayed a bias for the novel category of faces in relation to their previous exposure to infant and child faces. In Experiment 2, this pattern of visual attention was reversed in infants presented with pictures of personally familiar child faces (i.e., older siblings) compared with unfamiliar infant faces, especially in older infants. These results suggest that allocation of attention for novelty can supersede familiarity biases for faces depending on experience and highlight that multiple factors drive infant visual behavior in responding to the social world.

Cognitive flexibility and parental education differentially predict implicit and explicit racial biases in bilingual children.

Prior research has suggested that bilingual children demonstrate reduced social bias relative to their monolingual peers. In particular, they exhibit less implicit bias against racial outgroups. However, the cognitive determinants of racial bias in bilingual children remain unclear. In the current study, relationships between racial bias and three cognitive factors (inhibitory control, cognitive flexibility, and perspective-taking ability), along with language proficiency and parental education, were investigated in a sample of bilingual preschoolers (N = 55). Children were bilingual learners of English and Mandarin. Results demonstrated that implicit bias was predicted by cognitive flexibility, independent of variation in inhibitory control, second language vocabulary, perspective taking, and parental education. In contrast, explicit bias was predicted by parental education alone and not by cognitive or linguistic factors. Findings suggest that increased cognitive flexibility, often thought to be an outgrowth of bilingual experience, may also be associated with a reduction in implicit bias. Findings are discussed in terms of specific mechanisms that may link cognitive factors, bilingualism, and racial bias.

Tracking Reactions of Asymmetric Organo-Osmium Transfer Hydrogenation Catalysts in Cancer Cells.

Most metallodrugs are prodrugs that can undergo ligand exchange and redox reactions in biological media. Here we have investigated the cellular stability of the anticancer complex [OsII [(η6 -p-cymene)(RR/SS-MePh-DPEN)] [1] (MePh-DPEN=tosyl-diphenylethylenediamine) which catalyses the enantioselective reduction of pyruvate to lactate in cells. The introduction of a bromide tag at an unreactive site on a phenyl substituent of Ph-DPEN allowed us to probe the fate of this ligand and Os in human cancer cells by a combination of X-ray fluorescence (XRF) elemental mapping and inductively coupled plasma-mass spectrometry (ICP-MS). The BrPh-DPEN ligand is readily displaced by reaction with endogenous thiols and translocated to the nucleus, whereas the Os fragment is exported from the cells. These data explain why the efficiency of catalysis is low, and suggests that it could be optimised by developing thiol resistant analogues. Moreover, this work also provides a new way for the delivery of ligands which are inactive when administered on their own.

A passive hutch-cooling system for achieving high thermal-stability operation at the Nanoprobe beamline, Diamond Light Source.

The development of low-emittance storage rings and the rapid developments in nano-optics and imaging techniques are leading to decreasing X-ray spot sizes and increasing requirements on the environmental and mechanical stability of beamline components. In particular, temperature stability in the experimental hutches is critical to minimize uncontrolled displacements caused by thermal expansion and ensure consistent performance. Here, the design and thermal performance of the experimental hutches of the Nanoprobe beamline at Diamond Light Source are described, where a standard deviation of the room temperature down to 0.017°C over extended periods is demonstrated. The rooms are kept at constant temperature using water-cooled radiant panels which line the ceiling and walls. Radiant panels are relatively common in high-end electron microscopy rooms, but this is the first demonstration of their use for fine temperature control in an X-ray hutch and may provide a useful basis for future upgrades at upcoming low-emittance sources.

X-ray tomography of cryopreserved human prostate cancer cells: mitochondrial targeting by an organoiridium photosensitiser.

The organoiridium complex Ir[(C,N)2(O,O)] (1) where C, N = 1-phenylisoquinoline and O,O = 2,2,6,6-tetramethyl-3,5-heptanedionate is a promising photosensitiser for Photo-Dynamic Therapy (PDT). 1 is not toxic to cells in the dark. However, irradiation of the compound with one-photon blue or two-photon red light generates high levels of singlet oxygen (1O2) (in Zhang et al. Angew Chem Int Ed Engl 56 (47):14898-14902 https://doi.org/10.1002/anie.201709082,2017), both within cell monolayers and in tumour models. Moreover, photo-excited 1 oxidises key proteins, causing metabolic alterations in cancer cells with potent antiproliferative activity. Here, the tomograms obtained by cryo-Soft X-ray Tomography (cryo-SXT) of human PC3 prostate cancer cells treated with 1, irradiated with blue light, and cryopreserved to maintain them in their native state, reveal that irradiation causes extensive and specific alterations to mitochondria, but not other cellular components. Such new insights into the effect of 1O2 generation during PDT using iridium photosensitisers on cells contribute to a detailed understanding of their cellular mode of action.

Face Processing in Infancy and Beyond: The Case of Social Categories.

Prior reviews of infant face processing have emphasized how infants respond to faces in general. This review highlights how infants come to respond differentially to social categories of faces based on differential experience, with a focus on race and gender. We examine six different behaviors: preference, recognition, scanning, category formation, association with emotion, and selective learning. Although some aspects of infant responding to face race and gender may be accounted for by traditional models of perceptual development, other aspects suggest the need for a broader model that links perceptual development with social and emotional development. We also consider how responding to face race and gender in infancy may presage responding to these categories beyond infancy and discuss how social biases favoring own-race and female faces are formed.

A developmental investigation of the other-race categorization advantage in a multiracial population: Contrasting social categorization and perceptual expertise accounts.

Most prior studies of the other-race categorization advantage have been conducted in predominantly monoracial societies. This limitation has left open the question of whether tendencies to more rapidly and accurately categorize other-race faces reflect social categorization (own-race vs. other-race) or perceptual expertise (frequent exposure vs. infrequent exposure). To address this question, we tested Malay and Malaysian Chinese children (9- and 10-year-olds) and adults on (a) own-race faces (i.e., Malay faces for Malay participants and Chinese faces for Malaysian Chinese participants), (b) high-frequency other-race faces (i.e., Chinese faces for Malay participants and Malay faces for Malaysian Chinese participants), and (c) low-frequency other-race faces (i.e., Caucasian faces). Whereas the other-race categorization advantage was in evidence in the accuracy data of Malay adults, other aspects of performance were supportive of either the social categorization or perceptual expertise accounts and were dependent on the race (Malay vs. Chinese) or age (child vs. adult) of the participants. Of particular significance is the finding that Malaysian Chinese children and adults categorized own-race Chinese faces more rapidly than high-frequency other-race Malay faces. Thus, in accord with a perceptual expertise account, the other-race categorization advantage seems to be more an advantage for racial categories of lesser experience regardless of whether these face categories are own-race or other-race.

Sensitivity to race in language comprehension in monolingual and bilingual infants.

Past studies suggest that monolingual and bilingual infants respond differently to race information in face discrimination and social learning tasks. In particular, bilingual infants have been shown to respond more similarly to own- and other-race individuals, in contrast to monolingual infants, who respond preferentially to own-race individuals. In the current study, we investigated monolingual and bilingual sensitivity to speaker race in spoken word recognition. Two-year-old infants were presented with spoken words in association with visual targets. Words were presented in association with own- or other-race actors and were either correctly pronounced or mispronounced. Measuring speech-responsive eye movements to visual targets, we analyzed fixation to visual targets for correct and mispronounced words in relation to speaker race for each group. When presented with own-race speakers, both monolingual and bilingual infants associated correctly pronounced labels, but not mispronounced labels, with visual targets. When presented with other-race speakers, bilingual infants responded similarly. In contrast, monolingual infants did not fixate visual targets regardless of whether words were correctly pronounced or mispronounced by an other-race speaker. Results are discussed in terms of the sensitivities of bilingual and monolingual infants to novelty, learned associations between race and language, and prior social experiences.

Monolingual but not bilingual infants demonstrate racial bias in social cue use.

Bilingualism exerts early and pervasive effects on cognition, observable in infancy. Thus far, investigations of infant bilingual cognition have focused on sensitivity to visual memory, executive function, and linguistic sensitivity. Much less research has focused on how bilingualism impacts processing of social cues. The present study sought to investigate whether bilingualism modulates the expression of one aspect of social processing: early racial bias. Using a gaze-following paradigm, we investigated whether 18- to 20-month-old monolingual and bilingual infants favored their own race. Results demonstrated that monolingual infants favored their own race in following a model whose direction of gaze signaled an event. In contrast, bilingual infants demonstrated race-neutral gaze-following patterns, relying more heavily on the reliability of the behavior of the model over race. Findings suggest that bilingualism may have protective effects against the early emergence of racial bias.