Search

Abstract Childhood abuse reduces hippocampal and amygdala volumes and impairs social cognition, including the ability to recognize facial expressions. However, these associations have been studied primarily in individuals with a history of severe abuse and psychiatric symptoms; researchers have not determined whether these associations can also be observed in healthy adults. In the present study, we analyzed data from 400 healthy adults (208 men and 192 women) at Tamagawa University. Parental rejection reflecting childhood abuse was assessed using the short form of Egna Minnen Beträffande Uppfostran, while social cognition was assessed using the “Fake Smile Detection Task.” Hippocampal and amygdala volumes were extracted from T1-weighted magnetic resonance imaging data using FreeSurfer. We found that greater parental rejection resulted in smaller hippocampal and amygdala volumes and poorer performance in the Fake Smile Detection Task. Structural equation modeling analysis supported the model that hippocampal volume mediates maternal rejection effect on performance on the Fake Smile Detection Task, with involvement of the amygdala. These findings are in line with the structural and functional connectivity found between the hippocampus and amygdala and their joint involvement in social cognition. Therefore, parental rejection may affect hippocampal and amygdala volumes and social cognitive function even in symptom-free adults.

Abstract Prokaryotic communities inhabiting surface waters of temperate areas exhibit patterns of seasonal succession. Generally, studies describing these temporal changes are not performed in the proximity to the coast. In the present study, temporal variation of these communities was determined in surface waters at two stations located in the close proximity to the eastern shore of the northern Adriatic Sea. Sequencing of the V4 region of the 16S rRNA gene identified the highest community richness in December with distinct shifts in community structure between periods from April to May, June to October, and November to March. Temperature was shown to be the main environmental force explaining community temporal variation. The NS5 marine group, uncultured Cryomorphaceae, SAR86 clade, and Synechococcus were present throughout the year. Members without know relatives within Rhodobacteraceae and the NS4 marine group were more pronounced in the period from April to May, the AEGEAN-169 marine group, SAR11 subclade III, and HIMB11 in the period from June to October, and SAR11 subclade Ia and Archaea in the period from November to March. Litoricola and OM60 (NOR5) clade were characteristic for both the community sampled from April to May and November to March. Taken together, prokaryotic communities inhabiting nearshore surface waters exhibit a general pattern in community structure similar to other surface associated assemblages of temperate areas. However, the identified specific community composition and temporal patterns differ from other coastal areas.

Summary: Grid cells and place cells represent the spatiotemporal continuum of an animal’s past, present, and future locations. However, their spatiotemporal relationship is unclear. Here, we co-record grid and place cells in freely foraging rats. We show that average time shifts in grid cells tend to be prospective and are proportional to their spatial scale, providing a nearly instantaneous readout of a spectrum of progressively increasing time horizons ranging hundreds of milliseconds. Average time shifts of place cells are generally larger compared to grid cells and also increase with place field sizes. Moreover, time horizons display nonlinear modulation by the animal’s trajectories in relation to the local boundaries and locomotion cues. Finally, long and short time horizons occur at different parts of the theta cycle, which may facilitate their readout. Together, these findings suggest that population activity of grid and place cells may represent local trajectories essential for goal-directed navigation and planning.

Long-term memory tests are commonly used to facilitate the diagnosis of hippocampal-related neurological disorders such as Alzheimer's disease due to their relatively high specificity and sensitivity to damage to the medial temporal lobes compared to standard commonly used clinical tests. Pathological changes in Alzheimer's disease start years before the formal diagnosis is made, partially due to testing too late. This proof-of-concept exploratory study aimed to assess the feasibility of introducing an unsupervised digital platform for continuous testing of long-term memory over long periods outside the laboratory environment. To address this challenge, we developed a novel digital platform, hAge ('healthy Age'), which integrates double spatial alternation, image recognition and visuospatial tasks for frequent remote unsupervised assessment of spatial and non-spatial long-term memory carried out continuously over eight week period. To demonstrate the feasibility of our approach, we tested whether we could achieve sufficient levels of adherence and whether the performance on hAge tasks is comparable to the performance observed in the analogous standard tests measured in the controlled laboratory environments.191 healthy adults (67% females, 18-81 years old) participated in the study. We report an estimated 42.4% adherence level with minimal inclusion criteria. In line with findings using standard laboratory tests, we showed that performance on the spatial alternation task negatively correlated with inter-trial periods and the performance levels on image recognition and visuospatial tasks could be controlled by varying image similarity. Importantly, we demonstrated that frequent engagement with the double spatial alternation task leads to a strong practice effect, previously identified as a potential measure of cognitive decline in MCI patients. Finally, we discuss how lifestyle and motivation confounds may present a serious challenge for cognitive assessment in real-world uncontrolled environments.

The presence of seagrass shapes surface sediments and forms a specific environment for diverse and abundant microbial communities. A severe decline of Cymodocea nodosa, a widespread seagrass species in the Mediterranean Sea, has been documented. To characterise and assess the changes in microbial community composition during the decline of a Cymodocea nodosa meadow, Illumina MiSeq sequencing of the V4 region of the 16S rRNA gene was performed. Samples of surface sediments from two sites, one without any vegetation and one with a declining Cymodocea nodosa meadow, were collected at monthly intervals from July 2017 to October 2018. Microbial communities were stratified by sediment depth and differed between the vegetated and the nonvegetated site. Although the Cymodocea nodosa meadow declined to a point where almost no leaves were present, no clear temporal succession in the community was observed. Taxonomic analysis revealed a dominance of bacterial over archaeal sequences, with most archaeal reads classified as Nanoarchaeota, Thermoplasmatota, Crenarchaeota, and Asgardarchaeota. The bacterial community was mainly composed of Desulfobacterota, Gammaproteobacteria, Bacteroidota, Chloroflexi, Planctomycetota, and Campylobacterota. Our results show that sediment microbial communities are remarkably stable and may resist major disturbances such as seagrass meadow decline.

Abstract We have a keen sensitivity when it comes to the perception of our own voices. We can detect not only the differences between ourselves and others, but also slight modifications of our own voices. Here, we examined the neural correlates underlying such sensitive perception of one’s own voice. In the experiments, we modified the subjects’ own voices by using five types of filters. The subjects rated the similarity of the presented voices to their own. We compared BOLD (Blood Oxygen Level Dependent) signals between the voices that subjects rated as least similar to their own voice and those they rated as most similar. The contrast revealed that the bilateral superior temporal gyrus exhibited greater activities while listening to the voice least similar to their own voice and lesser activation while listening to the voice most similar to their own. Our results suggest that the superior temporal gyrus is involved in neural sharpening for the own-voice. The lesser degree of activations observed by the voices that were similar to the own-voice indicates that these areas not only respond to the differences between self and others, but also respond to the finer details of own-voices.

Surfaces of marine macrophytes are inhabited by diverse microbial communities. Most studies focusing on epiphytic communities of macrophytes did not take into account temporal changes or applied low sampling frequency approaches. The seasonal dynamics of epiphytic microbial communities was determined in a meadow of Cymodocea nodosa invaded by Caulerpa cylindracea and in a monospecific settlement of C. cylindracea at monthly intervals. For comparison the ambient prokaryotic picoplankton community was also characterized. At the OTU level, the microbial community composition differed between the ambient water and the epiphytic communities exhibiting host-specificity. Also, successional changes were observed connected to the macrophyte growth cycle. Taxonomic analysis, however, showed similar high rank taxa (phyla and classes) in the ambient water and the epiphytic communities, with the exception of Desulfobacterota, which were only found on C. cylindracea. Cyanobacteria showed seasonal changes while other high rank taxa were present throughout the year. In months of high Cyanobacteria presence the majority of cyanobacterial sequences were classified as Pleurocapsa. Phylogenetic groups present throughout the year (e.g., Saprospiraceae, Rhodobacteraceae, members without known relatives within Gammaproteobacteria, Desulfatitalea, and members without known relatives within Desulfocapsaceae) constituted most of the sequences, while less abundant taxa showed seasonal patterns connected to the macrophyte growth cycle. Taken together, epiphytic microbial communities of the seagrass C. nodosa and the macroalga C. cylindracea appear to be host-specific and contain taxa that undergo successional changes.

Th e purpose of the present study was to investigate the eff ects of exponential and linear tapering protocols on the distance covered according to playing positions among junior elite soccer players. One-hundred and fi ft y-eight junior elite soccer players (mean age: 17.07±0.79 years; mean height: 177.85±6.64 cm; mean weight: 71.27±7.96 kg; mean body-mass index: 22.50±1.66 kg/m²) were randomly selected into two groups: (1) exponential and (2) linear tapering group. Training sessions were conducted three times a week for eight weeks. Aft er four and eight weeks of the tapering period, participants were measured in the distance covered according to four basic playing positions on the fi eld: (1) goalkeepers, (2) defenders, (3) midfi elders, and (4) forwards. Th e results showed that the largest eff ects between the pre- and post-measurement had defenders in the exponential and goalkeepers in the linear group, goalkeepers in the distance covered by slow and medium running in both groups, forwards in fast running and sprinting in the exponential group, and midfi elders in the linear group. In conclusion, our results suggest that the exponential tapering protocol brought somewhat larger eff ects, especially in fast running and sprinting in forwards and slow and medium running in goalkeepers. Future studies need to implement both the exponential and the linear tapering protocols in order to enhance performance but favouring the exponential to produce larger eff ects.

Studies of unculturable microbes often combine methods, such as 16S rRNA sequencing, metagenomics, and metaproteomics. To apply these techniques to the microbial community inhabiting the surfaces of marine macrophytes, it is advisable to perform a selective DNA and protein isolation prior to the analysis to avoid biases due to the host material being present in high quantities. Two protocols for DNA and protein isolation were adapted for selective extractions of DNA and proteins from epiphytic communities inhabiting the surfaces of two marine macrophytes, the seagrass Cymodocea nodosa and the macroalga Caulerpa cylindracea. Protocols showed an almost complete removal of the epiphytic community regardless of the sampling season, station, settlement, or host species. The obtained DNA was suitable for metagenomic and 16S rRNA sequencing, while isolated proteins could be identified by mass spectrometry. Low presence of host DNA and proteins in the samples indicated a high specificity of the protocols. The procedures are based on universally available laboratory chemicals making the protocols widely applicable. Taken together, the adapted protocols ensure an almost complete removal of the macrophyte epiphytic community. The procedures are selective for microbes inhabiting macrophyte surfaces and provide DNA and proteins applicable in 16S rRNA sequencing, metagenomics, and metaproteomics.

The effect of the presence of invasive macroalgae Caulerpa cylindracea Sonder in the seagrass meadow Cymodocea nodosa (Ucria) Ascherson was studied by comparing the dynamics of biological and physicochemical parameters in the tissues of these two macrophytes and in the sediments underlying an invaded meadow (mixed settlement) and a C. cylindracea monospecific settlement. The study was conducted for 15 months, encompassing two summers during 2017 and 2018 (July to September) when maximum C. nodosa growth occurs. During 2017 C. cylindracea biomasses in the mixed settlement (79.5 ± 28.2 to 264.6 ± 65.1 g m–2) were lower than in its monospecific stands (113.4 ± 48.0 to 399.3 ± 56.3 g m–2). In the same period, less reducing conditions in the sediment underlying the mixed settlement were indicated by deeper redox transition depths (RTD: 8–12 mm) than those observed below the C. cylindracea monospecific settlement (RTD: 4–7 mm). In June 2018, C. cylindracea proliferated in both settlements reaching very similar biomasses that were maintained until September 2018 (mixed: 131.5 ± 23.0 to 172.5 ± 16.3 g m–2; monospecific: 162.8 ± 32.5 to 178.8 ± 30.0 g m–2). In parallel, a considerable lowering of RTD (5–7 mm) under the mixed settlement indicated the progression of stronger reducing conditions similar to those observed under the monospecific settlement (RTD: 0–7 mm). This alteration was followed by a decrease in C. nodosa below-ground biomass (89.3 ± 16.0 to 140.3 ± 24.3 g m–2), that became considerably lower than in the same period of 2017 (242.3 ± 44.3 to 346.9 ± 32.1 g m–2). At the same time, the above-ground biomass of C. nodosa (72.3 ± 14.8 to 110.3 ± 13.4 g m–2) showed no difference to the summer of 2017 (69.0 ± 15.4 to 116.0 ± 37.4 g m–2). The resulting increase of the above- to below-ground biomass ratio indicated the disruption of the meadow stability. More intense spawning of C. cylindracea in the mixed settlement during the summer 2018 hindered its expected proliferation in October 2018, while the below-ground biomass of C. nodosa increased concomitantly with the deepening of the RTD suggesting a possible recovery of the meadow stability.

Mammalian oocyte quality degrades over time after ovulation in vitro, which can cause fatal defects such as chromosomal aneuploidy. As various oocyte manipulations employed in assisted reproductive technology are time consuming, post-ovulatory aging is a serious problem to overcome in reproductive medicine or ova research. In this study, we investigated the effects of postovulatory aging on the incidence of chromosome aneuploidy during meiosis II, with a focus on the expression of functional proteins from the spindle assembly checkpoint (SAC). Chromosome analysis was used to assess the rate of aneuploidy in in vitro aged oocytes, or in early embryos derived from aged oocytes. Immunofluorescent staining was used to detect the localization of MAD2, which is a SAC signal that monitors the correct segregation of sister chromatids. Immunoblotting was used to quantify cohesin subunits, which are adhesion factors connecting sister chromatids at the metaphase II (MII) centromere. It was shown that post-ovulatory oocyte aging inhibits MAD2 localization to the sister kinetochore. Furthermore, oocyte aging prevented cohesin subunits from being maintained or degraded at the appropriate time. These data suggest that the destabilization of SAC signaling causes sister chromatid segregation errors in MII oocytes, and consequently increases the incidence of aneuploidy in early embryos. Our findings have provided distinct evidence that the post-ovulatory aging of oocytes might also be a risk factor for aneuploidy, irrespective of maternal age.

Three metagenomic libraries were constructed using surface sediment samples from the northern Adriatic Sea. Two of the samples were taken from a highly polluted and an unpolluted site respectively. The third sample from a polluted site had been enriched using crude oil. The results of the metagenome analyses were incorporated in the REDPET relational database (http://redpet.bioinfo.pbf.hr/REDPET), which was generated using the previously developed MEGGASENSE platform. The database includes taxonomic data to allow the assessment of the biodiversity of metagenomic libraries and a general functional analysis of genes using hidden Markov model (HMM) profiles based on the KEGG database. A set of 22 specialised HMM profiles was developed to detect putative genes for hydrocarbon-degrading enzymes. Use of these profiles showed that the metagenomic library generated after selection on crude oil had enriched genes for aerobic n-alkane degradation. The use of this system for bioprospecting was exemplified using potential alkB and almA genes from this library.

People perceive their recorded voice differently from their actively spoken voice. The uncanny valley theory proposes that as an object approaches humanlike characteristics, there is an increase in the sense of familiarity; however, eventually a point is reached where the object becomes strangely similar and makes us feel uneasy. The feeling of discomfort experienced when people hear their recorded voice may correspond to the floor of the proposed uncanny valley. To overcome the feeling of eeriness of own-voice recordings, previous studies have suggested equalization of the recorded voice with various types of filters, such as step, bandpass, and low-pass, yet the effectiveness of these filters has not been evaluated. To address this, the aim of experiment 1 was to identify what type of voice recording was the most representative of one's own voice. The voice recordings were presented in five different conditions: unadjusted recorded voice, step filtered voice, bandpass filtered voice, low-pass filtered voice, and a voice for which the participants freely adjusted the parameters. We found large individual differences in the most representative own-voice filter. In order to consider roles of sense of agency, experiment 2 investigated if lip-synching would influence the rating of own voice. The result suggested lip-synching did not affect own voice ratings. In experiment 3, based on the assumption that the voices used in previous experiments corresponded to continuous representations of non-own voice to own voice, the existence of an uncanny valley was examined. Familiarity, eeriness, and the sense of own voice were rated. The result did not support the existence of an uncanny valley. Taken together, the experiments led us to the following conclusions: there is no general filter that can represent own voice for everyone, sense of agency has no effect on own voice rating, and the uncanny valley does not exist for own voice, specifically.

The study investigates the control of structural response using collocated piezoelectric elements mounted on both sides of beam. A capacitive shunt is introduced into the circuitry to provide passive control of the beam’s configuration. The energy method is used to develop the structure’s governing equations. In order to select the appropriate electrostatic relations for the material, i.e. conductor or insulator material, the free charge rearrangement time is used. The effects of the capacitive shunt are introduced into the electrostatic relations using additive decomposition of capacitance. As an application of the model, a piezo-elastic beam restrictor using a capacitive shunt is investigated. Numerical results show that the capacitive shunt can effect passive control of the configuration of the beam.

Abstract Background As stereotactic body radiotherapy (SBRT) is a highly dose-dense radiotherapy, adverse events of neighboring normal tissues are a major concern. This study thus aimed to clarify the frequency and degree of clinical symptoms in patients with rib fractures after SBRT for primary lung cancer and to reveal risk factors for rib fracture. Appropriate α/β ratios for discriminating between fracture and non-fracture groups were also investigated. Methods Between November 2001 and April 2009, 177 patients who had undergone SBRT were evaluated for clinical symptoms and underwent follow-up thin-section computed tomography (CT). The time of rib fracture appearance was also assessed. Cox proportional hazard modeling was performed to identify risk factors for rib fracture, using independent variables of age, sex, maximum tumor diameter, radiotherapeutic method and tumor-chest wall distance. Dosimetric details were analyzed for 26 patients with and 22 randomly-sampled patients without rib fracture. Biologically effective dose (BED) was calculated with a range of α/β ratios (1–10 Gy). Receiver operating characteristics analysis was used to define the most appropriate α/β ratio. Results Rib fracture was found on follow-up thin-section CT in 41 patients. The frequency of chest wall pain in patients with rib fracture was 34.1% (14/41), and was classified as Grade 1 or 2. Significant risk factors for rib fracture were smaller tumor-chest wall distance and female sex. Area under the curve was maximal for BED at an α/β ratio of 8 Gy. Conclusions Rib fracture is frequently seen on CT after SBRT for lung cancer. Small tumor-chest wall distance and female sex are risk factors for rib fracture. However, clinical symptoms are infrequent and generally mild. When using BED analysis, an α/β ratio of 8 Gy appears most effective for discriminating between fracture and non-fracture patients.

Abstract Background Chest wall injury after stereotactic radiotherapy (SRT) for primary lung cancer has recently been reported. However, its detailed imaging findings are not clarified. So this study aimed to fully characterize the findings on computed tomography (CT), appearance time and frequency of chest wall injury after stereotactic radiotherapy (SRT) for primary lung cancer Materials and methods A total of 177 patients who had undergone SRT were prospectively evaluated for periodical follow-up thin-section CT with special attention to chest wall injury. The time at which CT findings of chest wall injury appeared was assessed. Related clinical symptoms were also evaluated. Results Rib fracture was identified on follow-up CT in 41 patients (23.2%). Rib fractures appeared at a mean of 21.2 months after the completion of SRT (range, 4 -58 months). Chest wall edema, thinning of the cortex and osteosclerosis were findings frequently associated with, and tending to precede rib fractures. No patients with rib fracture showed tumors > 16 mm from the adjacent chest wall. Chest wall pain was seen in 18 of 177 patients (10.2%), of whom 14 patients developed rib fracture. No patients complained of Grade 3 or more symptoms. Conclusion Rib fracture is frequently seen after SRT for lung cancer on CT, and is often associated with chest wall edema, thinning of the cortex and osteosclerosis. However, related chest wall pain is less frequent and is generally mild if present.

Multiple sclerosis (MS) is a progressive degenerative disorder of the central nervous system (CNS), characterized by inflammation, demyelination and axonal loss. While the majority of MS patients experience relapsing-remitting symptoms followed by a secondary progressive phase, about 10-15% patients exhibit a primary progressive disease involving continuous progression from its onset. Here we review the role of lectin-glycan recognition systems, including those concerning siglecs, C-type lectins and galectins in the pathogenesis of MS and experimental autoimmune encephalomyelitis. Particularly, we will focus on the role of galectins in the fate of T cells, dendritic cells and CNS cell populations. Understanding the regulatory circuits governed by lectin-glycan interactions and their association with disease-associated cytokine networks will contribute to develop novel therapeutic strategies in MS., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The present study was conducted to obtain a comprehensive overview of oligosaccharides present in the milk of a variety of important domestic animals including cows, goats, sheep, pigs, horses and dromedary camels. Using an analytical workflow that included ultra-performance liquid chromatography-hydrophilic interaction liquid chromatography with fluorescence detection coupled to quadrupole time-of-flight MS, detailed oligosaccharide libraries were established. The partial or full characterisation of the neutral/fucosylated, phosphorylated and sialylated structures was facilitated by sequencing with linkage- and sugar-specific exoglycosidases. Relative peak quantification of the 2-aminobenzamide-labelled oligosaccharides provided additional information. Milk from domestic animals contained a much larger variety of complex oligosaccharides than was previously assumed, and thirteen of these structures have been identified previously in human milk. The direct comparison of the oligosaccharide mixtures reflects their role in the postnatal maturation of different types of gastrointestinal systems, which, in this way, are prepared for certain post-weaning diets. The potential value of animal milk for the commercial extraction of oligosaccharides to be used in human and animal health is highlighted.

Aspergillus fumigatus is the causative agent of IA (invasive aspergillosis) in immunocompromised patients. It possesses a cell wall composed of chitin, glucan and galactomannan, polymeric carbohydrates synthesized by processive glycosyltransferases from intracellular sugar nucleotide donors. Here we demonstrate that A. fumigatus possesses an active AfAGM1 (A. fumigatus N-acetylphosphoglucosamine mutase), a key enzyme in the biosynthesis of UDP (uridine diphosphate)-GlcNAc (N-acetylglucosamine), the nucleotide sugar donor for chitin synthesis. A conditional agm1 mutant revealed the gene to be essential. Reduced expression of agm1 resulted in retarded cell growth and altered cell wall ultrastructure and composition. The crystal structure of AfAGM1 revealed an amino acid change in the active site compared with the human enzyme, which could be exploitable in the design of selective inhibitors. AfAGM1 inhibitors were discovered by high-throughput screening, inhibiting the enzyme with IC50s in the low μM range. Together, these data provide a platform for the future development of AfAGM1 inhibitors with antifungal activity.

Helicobacter pylori and Campylobacter jejuni colonize the stomach and intestinal mucus, respectively. Using a combination of mucus-secreting cells, purified mucins, and a novel mucin microarray platform, we examined the interactions of these two organisms with mucus and mucins. H. pylori and C. jejuni bound to distinctly different mucins. C. jejuni displayed a striking tropism for chicken gastrointestinal mucins compared to mucins from other animals and preferentially bound mucins from specific avian intestinal sites (in order of descending preference: the large intestine, proximal small intestine, and cecum). H. pylori bound to a number of animal mucins, including porcine stomach mucin, but with less avidity than that of C. jejuni for chicken mucin. The strengths of interaction of various wild-type strains of H. pylori with different animal mucins were comparable, even though they did not all express the same adhesins. The production of mucus by HT29-MTX-E12 cells promoted higher levels of infection by C. jejuni and H. pylori than those for the non-mucus-producing parental cell lines. Both C. jejuni and H. pylori bound to HT29-MTX-E12 mucus, and while both organisms bound to glycosylated epitopes in the glycolipid fraction of the mucus, only C. jejuni bound to purified mucin. This study highlights the role of mucus in promoting bacterial infection and emphasizes the potential for even closely related bacteria to interact with mucus in different ways to establish successful infections.

Carbohydrate structures play important roles in many biological processes, including cell adhesion, cell-cell communication, and host-pathogen interactions. Sugar nucleotides are activated forms of sugars used by the cell as donors for most glycosylation reactions. Using a liquid chromatography-tandem mass spectrometry-based method, we identified and quantified the pools of UDP-glucose, UDP-galactose, UDP-N-acetylglucosamine, GDP-mannose, and GDP-fucose in Plasmodium falciparum intraerythrocytic life stages. We assembled these data with the in silico functional reconstruction of the parasite metabolic pathways obtained from the P. falciparum annotated genome, exposing new active biosynthetic routes crucial for further glycosylation reactions. Fucose is a sugar present in glycoconjugates often associated with recognition and adhesion events. Thus, the GDP-fucose precursor is essential in a wide variety of organisms. P. falciparum presents homologues of GDP-mannose 4,6-dehydratase and GDP-L-fucose synthase enzymes that are active in vitro, indicating that most GDP-fucose is formed by a de novo pathway that involves the bioconversion of GDP-mannose. Homologues for enzymes involved in a fucose salvage pathway are apparently absent in the P. falciparum genome. This is in agreement with in vivo metabolic labeling experiments showing that fucose is not significantly incorporated by the parasite. Fluorescence microscopy of epitope-tagged versions of P. falciparum GDP-mannose 4,6-dehydratase and GDP-L-fucose synthase expressed in transgenic 3D7 parasites shows that these enzymes localize in the cytoplasm of P. falciparum during the intraerythrocytic developmental cycle. Although the function of fucose in the parasite is not known, the presence of GDP-fucose suggests that the metabolite may be used for further fucosylation reactions.

The enzymes phosphomannomutase (PMM), phospho-N-acetylglucosamine mutase (PAGM) and phosphoglucomutase (PGM) reversibly catalyse the transfer of phosphate between the C6 and C1 hydroxyl groups of mannose, N-acetylglucosamine and glucose respectively. Although genes for a candidate PMM and a PAGM enzymes have been found in the Trypanosoma brucei genome, there is, surprisingly, no candidate gene for PGM. The TbPMM and TbPAGM genes were cloned and expressed in Escherichia coli and the TbPMM enzyme was crystallized and its structure solved at 1.85 Å resolution. Antibodies to the recombinant proteins localized endogenous TbPMM to glycosomes in the bloodstream form of the parasite, while TbPAGM localized to both the cytosol and glycosomes. Both recombinant enzymes were able to interconvert glucose-phosphates, as well as acting on their own definitive substrates. Analysis of sugar nucleotide levels in parasites with TbPMM or TbPAGM knocked down by RNA interference (RNAi) suggests that, in vivo, PGM activity is catalysed by both enzymes. This is the first example in any organism of PGM activity being completely replaced in this way and it explains why, uniquely, T. brucei has been able to lose its PGM gene. The RNAi data for TbPMM also showed that this is an essential gene for parasite growth., (© 2012 Blackwell Publishing Ltd.)

Campylobacter jejuni is the leading cause of acute bacterial infectious diarrhea in humans. Unlike in humans, C. jejuni is a commensal within the avian host. Heavily colonized chickens often fail to display intestinal disease, and no cellular attachment or invasion has been demonstrated in-vivo. Recently, researchers have shown that the reason for the attenuation of C. jejuni virulence may be attributed to the presence of chicken intestinal mucus and more specifically chicken mucin. Since mucins are heavily glycosylated molecules this observation would suggest that glycan-based compounds may act as anti-infectives against C. jejuni. Considering this, we have investigated naturally sourced foods for potential anti-infective glycans. Bovine colostrum rich in neutral and acidic oligosaccharides has been identified as a potential source of anti-infective glycans. In this study, we tested oligosaccharides isolated and purified from the colostrum of Holstein Friesian cows for anti-infective activity against a highly invasive strain of C. jejuni. During our initial studies we structurally defined 37 bovine colostrum oligosaccharides (BCO) by HILIC-HPLC coupled with exoglycosidase digests and off-line mass spectroscopy, and demonstrated the ability of C. jejuni to bind to some of these structures, in-vitro. We also examined the effect of BCO on C. jejuni adhesion to, invasion of and translocation of HT-29 cells. BCO dramatically reduced the cellular invasion and translocation of C. jejuni, in a concentration dependent manner. Periodate treatment of the BCO prior to inhibition studies resulted in a loss of the anti-infective activity of the glycans suggesting a direct oligosaccharide-bacterial interaction. This was confirmed when the BCO completely prevented C. jejuni binding to chicken intestinal mucin, in-vitro. This study builds a strong case for the inclusion of oligosaccharides sourced from cow's milk in functional foods. However, it is only through further understanding the structure and function of milk oligosaccharides that such compounds can reach their potential as food ingredients., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Many studies have demonstrated the capacity of glycan-based compounds to disrupt microbial binding to mucosal epithelia. Therefore, oligosaccharides have potential application in the prevention of certain bacterial diseases. However, current screening methods for the identification of anti-adhesive oligosaccharides have limitations: they are time-consuming and require large amounts of oligosaccharides. There is a need to develop analytical techniques which can quickly screen for, and structurally define, anti-adhesive oligosaccharides prior to using human cell line models of infection. Considering this, we have developed a rapid method for screening complex oligosaccharide mixtures for potential anti-adhesive activity against bacteria. Our approach involves the use of whole bacterial cells to "deplete" free oligosaccharides from solution. As a case study, the free oligosaccharides from the colostrum of Holstein Friesian cows were screened for interactions with whole Escherichia coli cells. Reductions in oligosaccharide concentrations were determined by High pH Anion Exchange Chromatography and Hydrophilic Interaction Liquid Chromatography (HILIC-HPLC). Oligosaccharide structures were confirmed by a combination of HILIC-HPLC, exoglycosidase digestion and off-line negative ion mode MS/MS. The depletion assay confirmed selective bacterial interaction with certain bovine oligosaccharides which in previous studies, by other methodologies, had been shown to interact with E. coli. In particular, the bacterial cells depleted the following oligosaccharides in a population dependent manner: 3'-sialyllactose, disialyllactose, and 6'-sialyllactosamine. The assay methodology was further validated by studies in which we demonstrated the inhibitory activity of 3'-sialyllactose, and a mixture of bovine colostrum oligosaccharides, on E. coli adhesion to differentiated HT-29 cells., (Copyright © 2012 Elsevier B.V. All rights reserved.)