Giardia duodenalis is one of the most common causes of waterborne disease worldwide, and is often associated with outbreaks of diarrhea in areas with poor sanitation and hygiene. This study aimed to assess the prevalence and genetic diversity of G. duodenalis assemblages in individuals attending major public hospitals in Shiraz, southwestern Iran. From August 2022 to May 2023, a total of 614 stool samples from individuals were collected and initially examined for G. duodenalis cysts using parasitological techniques, sucrose flotation, and microscopy. Microscopy-positive samples were validated by SSU-PCR amplification of the parasite DNA. A multilocus genotyping (MLG) scheme, which focused on the triose phosphate isomerase (tpi) and the glutamate dehydrogenase (gdh) genes, was employed for genotyping purposes. G. duodenalis cysts were found in 7.5% (46/614) and 8.5% (52/614) of samples through microscopy and SSU-PCR, respectively. Successful amplification and sequencing results were obtained for 77.3% (17/22) and 45.5% (10/22) of the infected samples at the tpi and gdh loci, respectively. MLG data for the two loci were available for only five samples. Out of the 22 samples genotyped at any loci, 54.5% (12/22) were identified as assemblage A, while 45.5% (10/22) were identified as assemblage B. AII was the most predominant sub-assemblage identified [54.5% (12/22)], followed by BIII [27% (6/22)], discordant BIII/BIV [13.6% (3/22)], and BIV [4.5% (1/22)]. In the present study, no assemblages suited for non-human animal hosts (e.g., C-F) were detected. This suggests that the transmission of human giardiasis in Shiraz is primarily anthroponotic. Further molecular-based analyses are necessary to confirm and expand upon these findings. These analyses will also help determine the presence and public health importance of the parasite in environmental samples, such as drinking water.

Prokaryotes are known to produce and secrete a broad range of biopolymers with a high functional and structural heterogeneity, often with critical duties in the bacterial physiology and ecology. Among these, exopolysaccharides (EPS) play relevant roles in the interaction of bacteria with eukaryotic hosts. EPS can help to colonize the host and assist in bacterial survival, making this interaction more robust by facilitating the formation of structured biofilms. In addition, they are often key molecules in the specific recognition mechanisms involved in both beneficial and pathogenic bacteria-host interactions. A novel EPS known as MLG (Mixed-Linkage β-Glucan) was recently discovered in rhizobia, where it participates in bacterial aggregation and biofilm formation and is required for efficient attachment to the roots of their legume host plants. MLG is the first and, so far, the only reported linear Mixed-Linkage β-glucan in bacteria, containing a perfect alternation of β (1 → 3) and β (1 → 4) bonds. A phylogenetic study of MLG biosynthetic genes suggests that far from being exclusive of rhizobia, different soil and plant-associated bacteria likely produce MLG, adding this novel polymer to the plethora of surface polysaccharides that help bacteria thrive in the changing environment and to establish successful interactions with their hosts.In this work, a quantification method for MLG is proposed. It relays on the hydrolysis of MLG by a specific enzyme (lichenase), and the subsequent quantification of the released disaccharide (laminaribiose) by the phenol-sulfuric acid method. The protocol has been set up and optimized for its use in 96-well plates, which makes it suitable for high-throughput screening (HTS) approaches. This method stands out by its fast processing, technical simplicity, and capability to handle multiple samples and biological replicates at a time.

The prevalence and genetic diversity of the protozoan pathogen Giardia duodenalis have been extensively studied worldwide. There is currently a lack of data regarding the genetic variability of the organism in eastern India. Understanding the circulating genotypes and associated risk factors is crucial for effective planning and implementing control measures. Therefore, the objective of the study was to conduct an epidemiological study to determine the prevalence and identify the various genotypes present. This survey adds to our knowledge on the occurrence and distribution of Giardia genotypes in the studied region. The overall prevalence was found to be 6.8%. This parasitic infection was significantly associated with two age groups, i.e., >0-5 years and >5-12 years. Using a multilocus genotyping method, we genotyped 52 human Giardia isolates that were obtained from diarrheal patients. Two distinct assemblages were found in the population-30.8% belonged to assemblage A; 63.5% belonged to assemblage B, prevalent in the population; and 5.7% belonged to a combined assemblage A+B. Sub-assemblage AII was found in 17.3% of the cases, followed by sub-assemblage AI (13.5%). High levels of genetic diversity were found within the population of assemblage B undergoing balancing selection. Overall, the high prevalence of the parasite observed, particularly among children, raises a major concern and necessitates implementation of robust control measures. Furthermore, we report the presence of numerous unique genotypes, circulating in this limited geographical boundary, which can be useful dataset for future studies.

Pressure sensors urgently need high-performance sensing materials in order to be developed further. Sensitivity and creep are regarded as two key indices for assessing a sensor\'s performance. For the design and optimization of sensing materials, an accurate estimation of the impact of several parameters on sensitivity and creep is essential. In this study, sensitivity and creep were predicted using the response surface methodology (RSM) and support vector regression (SVR), respectively. The input parameters were the concentrations of nickel (Ni) particles, multiwalled carbon nanotubes (MWCNTs), and multilayer graphene (MLG), as well as the magnetic field intensity (B). According to statistical measures, the SVR model exhibited a greater level of predictability and accuracy. The non-dominated sorting genetic-II algorithm (NSGA-II) was used to generate the Pareto-optimal fronts, and decision-making was used to determine the final optimal solution. With these conditions, the optimized results revealed an improved performance compared to the earlier study, with an average sensitivity of 0.059 kPa-1 in the pressure range of 0-16 kPa and a creep of 0.0325, which showed better sensitivity in a wider range compared to previous work. The theoretical sensitivity and creep were relatively similar to the actual values, with relative deviations of 0.317% and 0.307% after simulation and experimental verification. Future research for transducer performance optimization can make use of the provided methodology because it is representative.

In this investigation, p-Mg2Si/n-Si heterojunction photodetector (PD) is fabricated by magnetron sputtering and low vacuum annealing in the absence of argon or nitrogen atmosphere. Multilayer Graphene (MLG)/Mg2Si/Si heterojunction PD is first fabricated by transferring MLG to Mg2Si/Si heterojunction substrate using the suspended self-help transfer MLG method. After characterizing the phase composition, morphology and detection properties of Mg2Si/Si and MLG/Mg2Si/Si heterojunction PDs, the successful fabrication of the Mg2Si/Si and MLG/Mg2Si/Si heterojunction PDs are confirmed and some detection capabilities are realized. Compared with the Mg2Si/Si heterojunction PD, the light absorption and the ability to effectively separate and transfer photogenerated carriers of MLG/Mg2Si/Si heterojunction PD are improved. The responsivity, external quantum efficiency (EQE), noise equivalent power (NEP), detectivity (D*), on/off ratio and other detection properties are enhanced. The peak responsivity and EQE of the MLG/Mg2Si/Si heterojunction PD are 23.7 mA/W and 2.75%, respectively, which are better than the previous 1-10 mA/W and 2.3%. The results illustrate that the fabrication technology of introducing MLG to regulate the detection properties of the Mg2Si/Si heterojunction PD is feasible. In addition, this study reveals the potential of MLG to enhance the detection properties of optoelectronic devices, broadens the application prospect of the Mg2Si/Si-based heterojunction PDs and provides a direction for the regulation of optoelectronic devices.

Barley cultivation is adversely affected by high-temperature stress, which may modulate plant defense responses to pathogens such as barley powdery mildew (Blumeria graminis f. sp. hordei, Bgh). Earlier research focused mainly on the influence of short-term heat stress (heat shock) of barley on Bgh infection. In this study, our aim was to investigate the effects of both short- and long-term heat stress (35 °C from 30 s to 5 days) on Bgh infection in the barley cultivar Ingrid and its near-isogenic lines containing different powdery mildew resistance genes (Mla12, Mlg, and mlo5) by analyzing symptom severity and Bgh biomass with RT-qPCR. The expression of selected barley defense genes (BAX inhibitor-1, Pathogenesis- related protein-1b, Respiratory burst oxidase homologue F2, and Heat shock protein 90-1) was also monitored in plants previously exposed to heat stress followed by inoculation with Bgh. We demonstrated that pre-exposure to short- and long-term heat stress negatively affects the resistance of all resistant lines manifested by the appearance of powdery mildew symptoms and increased Bgh biomass. Furthermore, prolonged heat stress (48 and 120 h) enhanced both Bgh symptoms and biomass in susceptible wild-type Ingrid. Heat stress suppressed and delayed early defense gene activation in resistant lines, which is a possible reason why resistant barley became partially susceptible to Bgh.

The glucan content of rice is a key factor defining its nutritional and economic value. Starch and its derivatives have many industrial applications such as in fuel and material production. Non-starch glucans such as (1,3;1,4)-β-D-glucan (mixed-linkage β-glucan, MLG) have many benefits in human health, including lowering cholesterol, boosting the immune system, and modulating the gut microbiome. In this study, the genetic variability of MLG and starch contents were analyzed in rice (Oryza sativa L.) whole grain, by performing a new quantitative analysis of the polysaccharide content of rice grains. The 197 rice accessions investigated had an average MLG content of 252 μg/mg, which was negatively correlated with the grain starch content. A new genome-wide association study revealed seven significant quantitative trait loci (QTLs) associated with the MLG content and two QTLs associated with the starch content in rice whole grain. Novel genes associated with the MLG content were a hexose transporter and anthocyanidin 5,3-O-glucosyltransferase. Also, the novel gene associated with the starch content was a nodulin-like domain. The data pave the way for a better understanding of the genes involved in determining both MLG and starch contents in rice grains and should facilitate future plant breeding programs.

Plant cell walls (PCWs) form the outer barrier of cells that give the plant strength and directly interact with the environment and other cells in the plant. PCWs are composed of several polysaccharides, of which cellulose forms the main fibrillar network. Enmeshed between these fibrils of cellulose are non-cellulosic polysaccharides (NCPs), pectins, and proteins. This study investigates the sequence, timing, patterning, and architecture of cell wall polysaccharide regeneration in suspension culture cells (SCC) of the grass species Lolium multiflorum (Lolium). Confocal, superresolution, and electron microscopies were used in combination with cytochemical labeling to investigate polysaccharide deposition in SCC after protoplasting. Cellulose was the first polysaccharide observed, followed shortly thereafter by (1,3;1,4)-β-glucan, which is also known as mixed-linkage glucan (MLG), arabinoxylan (AX), and callose. Cellulose formed fibrils with AX and produced a filamentous-like network, whereas MLG formed punctate patches. Using colocalization analysis, cellulose and AX were shown to interact during early stages of wall generation, but this interaction reduced over time as the wall matured. AX and MLG interactions increased slightly over time, but cellulose and MLG were not seen to interact. Callose initially formed patches that were randomly positioned on the protoplast surface. There was no consistency in size or location over time. The architecture observed via superresolution microscopy showed similarities to the biophysical maps produced using atomic force microscopy and can give insight into the role of polysaccharides in PCWs.

Cell walls of marine macroalgae are composed of diverse polysaccharides that provide abundant carbon sources for marine heterotrophic bacteria. Among them, Zobellia galactanivorans is considered as a model for studying algae-bacteria interactions. The degradation of typical algal polysaccharides, such as agars or alginate, has been intensively studied in this model bacterium, but the catabolism of plant-like polysaccharides is essentially uncharacterized. Here, we identify a polysaccharide utilization locus in the genome of Z. galactanivorans, induced by laminarin (β-1,3-glucans), and containing a putative GH5 subfamily 4 (GH5_4) enzyme, currently annotated as a endoglucanase (ZgEngAGH5_4). A phylogenetic analysis indicates that ZgEngAGH5_4 was laterally acquired from an ancestral Actinobacteria We performed the biochemical and structural characterization of ZgEngAGH5_4 and demonstrated that this GH5 is, in fact, an endo-β-glucanase, most active on mixed-linked glucan (MLG). Although ZgEngAGH5_4 and GH16 lichenases both hydrolyze MLG, these two types of enzymes release different series of oligosaccharides. Structural analyses of ZgEngAGH5_4 reveal that all the amino acid residues involved in the catalytic triad and in the negative glucose-binding subsites are conserved, when compared with the closest relative, the cellulase EngD from Clostridium cellulovorans, and some other GH5s. In contrast, the positive glucose-binding subsites of ZgEngAGH5_4 are different and this could explain the preference for MLG, with respect to cellulose or laminarin. Molecular dynamics computer simulations using different hexaoses reveal that the specificity for MLG occurs through the +1 and +2 subsites of the binding pocket that display the most important differences when compared with the structures of other GH5_4 enzymes.

Giardia duodenalis (syn. Giardia lamblia, Giardia intestinalis) is an important zoonotic parasite that parasitizes the gastro-intestines of humans and animals, with diarrhea as the most common clinical symptom. The goat has been indicated as one of the most important reservoirs of G. duodenalis for humans. In the present study, we investigated the prevalence and genetic diversity of G. duodenalis in goats in Shaanxi province, northwestern China. A total of 1311 faecal specimens were examined, and the overall prevalence was 7.1% (93/1311). Although all the meat, cashmere and dairy goats were positive for infection, the highest prevalence was found in cashmere goats (10.2%), followed by dairy (9.4%) and meat goats (2.0%). Negative correlation between age and prevalence was also observed, and the highest prevalence was detected in 0-2-month goats. Genetic analysis showed the presence of three assemblages, including two zoonotic (A and B) and one animal-adapted assemblage E, with E as the prevalent assemblage found in all breeds of positive goats. The zoonotic assemblage A was found in Guanzhong dairy and Shanbei cashmere goats, but B was only detected in Boar goats. Additionally, mixed assemblages E and A were also identified in two cashmere goats. Multi-locus genotyping (MLST) using the gene loci of the triosephosphate isomerase (tpi), b-giardin (bg) and glutamate dehydrogenase (gdh) identified four novel multi-locus genotypes (MLGs), including two assemblage E MLGs and two assemblage A MLGs. These results suggested that Boar, Guanzhong dairy and Shanbei cashmere goats in Shaanxi province would be potential reservoirs for human infections in this area, and this study also provided basic data for controlling G. duodenalis infection in goats as well as other hosts.