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The current study investigates whether there are statistically independent age-related influences on the canine cognitive structure and how individual factors moderate cognitive aging on both cross-sectional and longitudinal samples. A battery of seven tasks was administered to 129 pet dogs, on which exploratory and confirmatory factor analyses were employed to unveil the correlational structure underlying individual differences in cognitive performance. The best-fitting model featured a hierarchical structure with two first-order cognitive domains (individual problem solving, learning) and a second-order common factor. These higher order factors exhibited consistency over a period of at least 2.5 years. External validation linked the common factor positively to discrimination and reversal learning performance, exploration, neophilia, activity/excitability, and training level while negatively to cognitive dysfunction symptoms, suggesting that it is a good candidate for a general cognitive factor (canine g). Structural equation models identified three distinct age-related influences, operating on associative learning, on memory, and on canine g. Health status moderated the negative age-canine g relationship, with a stronger association observed in dogs with poorer health status, and no relationship for dogs in good health. On a longitudinal sample (N = 99), we showed that the direction and magnitude of change in canine g over up to 3 years is affected by various interactions between the dogs' age, communication score, baseline performance, and time elapsed since the baseline measurement. These findings underscore the presence of a general cognitive factor in dogs and reveal intriguing parallels between human and canine aging, affirming the translational value of dogs in cognition and aging research., (© 2024. The Author(s).)

Efforts to counteract age-related decline have resulted in the emergence of various interventions. However, everyday benefits are rarely reported in elderly people. Dogs provide an excellent model for studying aging and interventions due to their similarities to humans. Our aim was to investigate whether a combined physical and cognitive intervention (most effective in humans) could enhance the performance of pet dogs and lead to far transfer effects (improvement in not just the trained specific task). We examined the impact of three-month-long intervention therapies (cognitive, physical, combined) on the cognitive performance and behaviour of old, healthy dogs (N = 72; aged 7.68-14.54 years) using a 12-subtest behavioural test battery. We did not find the combined intervention group outperforming either the cognitive-only or physical-only therapy groups. Physical interventions, either alone or in combination, improved dogs' behavioural flexibility and social behaviour. Cognitive interventions, either alone or in combination, increased neophilia. Furthermore, all intervention therapies made dogs more engaged with their environment. Moreover, less old, around eight years old dogs, exhibited improved social behaviour, problem solving ability, and increased neophilia by their second test occasion. Additionally, dogs' performance was influenced by their health, training, daily play with the owner, and activity/excitability traits. In sum, both cognitive and physical intervention therapies can have an impact on the behaviour of old, healthy pet dogs. However, these therapies may be more effective when longer or applied at a younger age, as the healthy older dogs were less likely to show improvement., (© 2024. The Author(s).)

Chemical dynamics in biological samples are seldom stand-alone processes but represent the outcome of complicated cascades of interlinked reaction chains. In order to understand these processes and how they correlate, it is important to monitor several parameters simultaneously at high spatial and temporal resolution. Hyperspectral imaging is a promising tool for this, as it provides broad-range spectral information in each pixel, enabling the use of multiple luminescent indicator dyes, while simultaneously providing information on sample structures and optical properties. In this study, we first characterized pH- and O 2 -sensitive indicator dyes incorporated in different polymer matrices as optical sensor nanoparticles to provide a library for (hyperspectral) chemical imaging. We then demonstrate the successful combination of a pH-sensitive indicator dye (HPTS(DHA) 3 ), an O 2 -sensitive indicator dye (PtTPTBPF), and two reference dyes (perylene and TFPP), incorporated in polymer nanoparticles for multiparameter chemical imaging of complex natural samples such as green algal biofilms ( Chlorella sorokiniana ) and seagrass leaves ( Zostera marina ) with high background fluorescence. We discuss the system-specific challenges and limitations of our approach and further optimization possibilities. Our study illustrates how multiparameter chemical imaging with hyperspectral read-out can now be applied on natural samples, enabling the alignment of several chemical parameters to sample structures.

Molecularly imprinted polymer (MIP) nanofilms for alpha-fetoprotein (AFP) and the receptor binding domain (RBD) of the spike protein of SARS-CoV-2 using either a peptide (epitope-MIP) or the whole protein (protein-MIP) as the template were prepared by electropolymerization of scopoletin. Conducting atomic force microscopy revealed after template removal and electrochemical deposition of gold a larger surface density of imprinted cavities for the epitope-imprinted polymers than when using the whole protein as template. However, comparable affinities towards the respective target protein (AFP and RBD) were obtained for both types of MIPs as expressed by the K D values in the lower nanomolar range. On the other hand, while the cross reactivity of both protein-MIPs towards human serum albumin (HSA) amounts to around 50% in the saturation region, the nonspecific binding to the respective epitope-MIPs is as low as that for the non-imprinted polymer (NIP). This effect might be caused by the different sizes of the imprinted cavities. Thus, in addition to the lower costs the reduced nonspecific binding is an advantage of epitope-imprinted polymers for the recognition of proteins., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Dogs interpret cues as being about location, which human infants would relate to objects. This spatial bias could shed light on the evolution of object-centered thought, however, research needs to rule out that this is not a by-product of dogs' weaker (compared to humans) visual capacities. In this study, we used a data set in which dogs were tested in two types of learning tasks (discrimination and reversal learning) with two types of rewarded cues (location and object features). In both tasks, dogs displayed spatial bias, that is, faster learning when the rewarded cue was a location. We investigated how sensory and cognitive capacity each contributes to this spatial bias. To this end, an estimate for general cognitive ability (g) was obtained from a battery of tests for some of the dogs. Cephalic index, a feature targeted in breeding and linked to differences in visual capacity, correlated negatively with the expression of spatial bias only in the easier discrimination learning task, while a negative correlation between g factor and spatial bias scores emerged in the more difficult reversal learning task. We conclude that dogs' spatial bias cannot be reduced to a sensory limitation and is easier to overcome with greater cognitive capacity., Competing Interests: Conflict of Interest Statement The authors declare no conflict of interest.

The shape of the cranium is one of the most notable physical changes induced in domestic dogs through selective breeding and is measured using the cephalic index (CI). High CI (a ratio of skull width to skull length > 60) is characterized by a short muzzle and flat face and is referred to as brachycephaly. Brachycephalic dogs display some potentially harmful changes in neuroanatomy, and there are implications for differences in behavior, as well. The path from anatomy to cognition, however, has not been charted in its entirety. Here, we report that sleep-physiological markers of white-matter loss (high delta power, low frontal spindle frequency, i.e., spindle waves/s), along with a spectral profile for REM (low beta, high delta) associated with low intelligence in humans, are each linked to higher CI values in the dog. Additionally, brachycephalic subjects spent more time sleeping, suggesting that the sleep apnea these breeds usually suffer from increases daytime sleepiness. Within sleep, more time was spent in the REM sleep stage than in non-REM, while REM duration was correlated positively with the number of REM episodes across dogs. It is currently not clear if the patterns of sleep and sleep-stage duration are mainly caused by sleep-impairing troubles in breathing and thermoregulation, present a juvenile-like sleeping profile, or are caused by neuro-psychological conditions secondary to the effects of brachycephaly, e.g., frequent REM episodes are known to appear in human patients with depression. While future studies should more directly address the interplay of anatomy, physiology, and behavior within a single experiment, this represents the first description of how the dynamics of the canine brain covary with CI, as measured in resting companion dogs using a non-invasive sleep EEG methodology. The observations suggest that the neuroanatomical changes accompanying brachycephaly alter neural systems in a way that can be captured in the sleep EEG, thus supporting the utility of the latter in the study of canine brain health and function., (© 2023. The Author(s).)

Despite serious health and longevity problems, small brachycephalic breeds are becoming increasingly popular among pet owners. Motivations for choosing short-nosed breeds have been extensively investigated in recent years; however, this issue has been addressed mainly by relying on owner reports, resulting in explanations of "cute looks", referring to the baby-schema phenomenon and "behaviour well suited for companionship". We aimed to compare the behaviour of two brachycephalic (English and French bulldogs) and one mesocephalic (Mudi) breed in a problem-solving context. The dogs were given the task of opening boxes containing food rewards. We investigated human-directed behaviour elements over success and latency (indicators of motivation and ability). We found that both English and French bulldogs were significantly less successful in solving the problem than mudis. Both brachycephalic breeds had longer opening latencies than the mesocephalic breed. Brachycephalic breeds oriented less at the problem box and more at humans present. In summary, the short-headed breeds were less successful but oriented much more toward humans than mesocephalic dogs. Owners might interpret these behaviours as "helplessness" and dependence. The results support the hypothesis that infant-like traits may be present not only in appearance but also in behaviour in brachycephalic breeds, eliciting caring behaviour in owners., (© 2023. Springer Nature Limited.)

Here we aim to gain a mechanistic understanding of the formation of epitope-imprinted polymer nanofilms using a non-terminal peptide sequence, i.e. the peptide GFNCYFP (G485 to P491) of the SARS-CoV-2 receptor binding domain (RBD). This epitope is chemisorbed on the gold surface through the central cysteine 488 followed by the electrosynthesis of a ∼5 nm thick polyscopoletin film around the surface confined templates. The interaction of peptides and the parent RBD and spike protein with the imprinted polyscopoletin nanofilm was followed by electrochemical redox marker gating, surface enhanced infrared absorption spectroscopy and conductive AFM. Because the use of non-terminal epitopes is especially intricate, here we characterize the binding pockets through their interaction with 5 peptides rationally derived from the template sequence, i.e. implementing central single amino acid mismatch as well as elongations and truncations at its C- and N- termini. Already a single amino acid mismatch, i.e. the central Cys488 substituted by a serine, results in ca. 15-fold lower affinity. Further truncation of the peptides to tetrapeptide (EGFN) and hexapeptide (YFPLQS) results also in a significantly lower affinity. We concluded that the affinity towards the different peptides is mainly determined by the four amino acid motif CYFP present in the sequence of the template peptide. A higher affinity than that for the peptides is found for the parent proteins RBD and spike protein, which seems to be due to out of cavity effects caused by their larger footprint on the nanofilm surface.

In situ labelling and encapsulation of biological entities, such as of single viruses, may provide a versatile approach to modulate their functionality and facilitate their detection at single particle level. Here, we introduce a novel virus metallization approach based on in situ coating of viruses in solution with silver nanoparticles (AgNP) in a two-step synthetic process, i.e. surface activation with a tannic acid - Sn(II) coordination complex, which subsequently induces silver ion (I) reduction. The metalic coating on the virus surface opens the opportunity for electrochemical quantification of the AgNP-tagged viruses by nano-impact electrochemistry on a microelectrode with single particle sensitivity, i.e. enable the detection of particles oherwise undetectable. We show that the silver coating of the virus particles impacting the electrode can be oxidized to produce distinct current peaks the frequency of which show a linear correlation with the virus count. The proof of the concept was done with inactivated Influenza A (H3N2) viruses resulting in their quantitation down to the femtomolar concentrations (ca. 5 × 10 7 particles per mL) using 50 s counting sequences.

We introduce a practically generic approach for the generation of epitope-imprinted polymer-based microarrays for protein recognition on surface plasmon resonance imaging (SPRi) chips. The SPRi platform allows the subsequent rapid screening of target binding kinetics in a multiplexed and label-free manner. The versatility of such microarrays, both as synthetic and screening platform, is demonstrated through developing highly affine molecularly imprinted polymers (MIPs) for the recognition of the receptor binding domain (RBD) of SARS-CoV-2 spike protein. A characteristic nonapeptide GFNCYFPLQ from the RBD and other control peptides were microspotted onto gold SPRi chips followed by the electrosynthesis of a polyscopoletin nanofilm to generate in one step MIP arrays. A single chip screening of essential synthesis parameters, including the surface density of the template peptide and its sequence led to MIPs with dissociation constants ( K D ) in the lower nanomolar range for RBD, which exceeds the affinity of RBD for its natural target, angiotensin-convertase 2 enzyme. Remarkably, the same MIPs bound SARS-CoV-2 virus like particles with even higher affinity along with excellent discrimination of influenza A (H3N2) virus. While MIPs prepared with a truncated heptapeptide template GFNCYFP showed only a slightly decreased affinity for RBD, a single mismatch in the amino acid sequence of the template, i.e. the substitution of the central cysteine with a serine, fully suppressed the RBD binding., Competing Interests: The authors declare no conflict of interest., (This journal is © The Royal Society of Chemistry.)

Forming eye contact is important in dog-human communication. In this study we measured what factors affect dogs' propensity for forming eye contact with an experimenter. We investigated the effect of [1] cephalic index (head shape's metric, indicator of higher visual acuity at the centre of the visual field), [2] breed function (visual cooperativeness), [3] age and [4] playfulness with strangers in 125 companion dogs. Cephalic index was measured individually and analysed as a continuous variable. Results showed that [1] dogs with a higher cephalic index (shorter head) established eye contact faster. Since cephalic index is highly variable even within a breed, using artificial head shape groups or breed average cephalic index values is not recommended. [2] Breed function also affected dogs' performance: cooperative breeds and mongrels established eye contact faster than dogs from non-cooperative breeds. [3] Younger dogs formed eye contact faster than older ones. [4] More playful dogs formed eye contact faster. Our results suggest that several factors affect dogs' interspecific attention, and therefore their visual communication ability.

Nucleic acid aptamers show clear promise as diagnostic reagents, as highly specific strands were reported against a large variety of biomarkers. They have appealing benefits in terms of reproducible generation by chemical synthesis, controlled modification with labels and functionalities providing versatile means for detection and oriented immobilization, as along with high biochemical and temperature resistance. Aptamers against immunoglobulin targets-IgA, IgM, IgG and IgE-have a clear niche for diagnostic applications, therefore numerous aptamers have been selected and used in combination with a variety of detection techniques. The aim of this review is to overview and evaluate aptamers selected for the recognition of antibodies, in terms of their design, analytical properties and diagnostic applications. Aptamer candidates showed convincing performance among others to identify stress and upper respiratory tract infection through SIgA detection, for cancer cell recognition using membrane bound IgM, to detect and treat hemolytic transfusion reactions, autoimmune diseases with IgG and detection of IgE for allergy diseases. However, in general, their use still lags significantly behind what their claimed benefits and the plethora of application opportunities would forecast.

Several studies on age-related cognitive decline in dogs involve laboratory dogs and prolonged training. We developed two spatial tasks that required a single 1-h session. We tested 107 medium-large sized dogs: "young" (N=41, aged 2.5-6.5 years) and "old" (N=66, aged 8-14.5 years). Our results indicated that, in a discrimination learning task and in a reversal learning task, young dogs learned significantly faster than the old dogs, indicating that these two tasks could successfully be used to investigate differences in spatial learning between young and old dogs. We also provide two novel findings. First, in the reversal learning, the dogs trained based on the location of stimuli learned faster than the dogs trained based on stimulus characteristics. Most old dogs did not learn the task within our cut-off of 50 trials. Training based on an object's location is therefore more appropriate for reversal learning tasks. Second, the contrast between the response to the positive and negative stimuli was narrower in old dogs, compared to young dogs, during the reversal learning task, as well as the cognitive bias test. This measure favors comparability between tasks and between studies. Following the cognitive bias test, we could not find any indication of differences in the positive and negative expectations between young and old dogs. Taken together, these findings do not support the hypothesis that old dogs have more negative expectations than young dogs and the use of the cognitive bias test in older dogs requires further investigation.

The gaze of other dogs and humans is informative for dogs, but it has not been explored which factors predict face-directed attention. We used image presentations of unfamiliar human and dog heads, facing the observer (portrait) or facing away (profile), and measured looking time responses. We expected dog portraits to be aversive, human portraits to attract interest, and tested dogs of different sex, skull length and breed function, which in previous work had predicted human-directed attention. Dog portraits attracted longer looking times than human profiles. Mesocephalic dogs looked at portraits longer than at profiles, independent of the species in the image. Overall, brachycephalic dogs and dogs of unspecified breed function (such as mixed breeds) displayed the longest looking times. Among the latter, females observed the images for longer than males, which is in line with human findings on sex differences in processing faces. In a subsequent experiment, we tested whether dog portraits functioned as threatening stimuli. We hypothesized that dogs will avoid food rewards or approach them more slowly in the presence of a dog portrait, but found no effect of image type. In general, older dogs took longer to approach food placed in front of the images and mesocephalic dogs were faster than dogs of other skull length types. The results suggest that short-headed dogs are more attentive to faces, while sex and breed function predict looking times through complex interactions.

Long-read genomic applications, such as genome mapping in nanochannels, require long DNA that is free of small-DNA impurities. We have developed a chip-based system based on entropic trapping that can simultaneously concentrate and purify a long DNA sample under the alternating application of an applied pressure (for sample injection) and an electric field (for filtration and concentration). In contrast, short DNA tends to pass through the filter owing to its comparatively weak entropic penalty for entering the nanoslit. The single-stage prototype developed here, which operates in a continuous pulsatile manner, achieves selectivities of up to 3.5 for λ-phage DNA (48.5 kilobase pairs) compared to a 2 kilobase pair standard based on experimental data for the fraction filtered using pure samples of each species. The device is fabricated in fused silica using standard clean-room methods, making it compatible for integration with long-read genomics technologies.

A Hungarian soldier previously immunized against Neisseria meningitidis by quadrivalent polysaccharide vaccine was twice infected with meningococci within six weeks. The patient was treated with ceftriaxone during both episodes and he successfully recovered. His close contacts received rifampicin prophylaxis. An investigation was performed to characterize the genetic background of the pathogens to ascertain if the recurrent invasive meningococcal disease was caused by the same strain and to find out the reason for reinfection. Both meningococci belonged to the fine type Y:P1.5-2,10-1:F4-1:ST-23. This is the first description of the Europe-wide prevalent N. meningitidis serogroup Y in Hungary. In the first episode, we found wild-type rpoB allele in the non-culturable sample implying the susceptibility to rifampicin. The culturable isolate from the second episode proved resistant to rifampicin and had a point mutation in the rpoB gene. The rifampicin resistance might have evolved during the prophylactic treatment of contacts. Previous immunization of the patient with polysaccharide vaccine was ineffective due to his immunodeficiency, thus immunization with conjugate vaccine was proposed. We have proposed the implementation of centralized rifampicin susceptibility testing of N. meningitidis strains within a defined time frame to intervene and administer appropriate prophylaxis to close contacts.

Hyponatremia is a potential side-effect of antidepressants that was observed most frequently in connection with compounds acting on the serotonin system. Risk of hyponatremia was found to be higher in the elderly. Authors report a case of a 65-year old male patient who was treated with venlafaxine-mirtazapine combination for recurrent depression. Severe hyponatremia, requiring intensive care, emerged in connection with the treatment. The authors emphasize the importance of the regular checking of serum sodium levels during antidepressant treatment, especially in elderly patients.