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The distinct conjugation machineries encoded by plasmids pNP40 and pUC11B represent the most prevalent plasmid transfer systems among lactococcal strains. In the current study, we identified genetic determinants that underpin pNP40‐ and pUC11B‐mediated, high‐frequency mobilisation of other, non‐conjugative plasmids. The mobilisation frequencies of the smaller, non‐conjugative plasmids and the minimal sequences required for their mobilisation were determined, owing to the determination of the oriT sequences of both pNP40 and pUC11B, which allowed the identification of similar sequences in some of the non‐conjugative plasmids that were shown to promote their mobilisation. Furthermore, the auxiliary gene mobC, two distinct functional homologues of which are present in several plasmids harboured by the pNP40‐ and pUC11B‐carrying host strains, was observed to confer a high‐frequency mobilisation phenotype. These findings provide mechanistic insights into how lactococcal conjugative plasmids achieve conjugation and promote mobilisation of non‐conjugative plasmids. Ultimately, these insights would be harnessed to optimise conjugation and mobilisation strategies for the rapid and predictable development of robust and technologically improved strains. [ABSTRACT FROM AUTHOR]

Bacteriophages (or phages) represent a persistent threat to the success and reliability of food fermentation processes. Recent reports of phages that infect Streptococcus thermophilus have highlighted the diversification of phages of this species. Phages of S. thermophilus typically exhibit a narrow range, a feature that is suggestive of diverse receptor moieties being presented on the cell surface of the host. Cell wall polysaccharides, including rhamnose-glucose polysaccharides and exopolysaccharides have been implicated as being involved in the initial interactions with several phages of this species. Following internalization of the phage genome, the host presents several defences, including CRISPR-Cas and restriction and modification systems to limit phage proliferation. This review provides a current and holistic view of the interactions of phages and their S. thermophilus host cells and how this has influenced the diversity and evolution of both entities. [ABSTRACT FROM AUTHOR]

[Display omitted] • An simulation-driven-design tool is developed to design metal lattices with a target compressive stress–strain curve. • The automated design algorithm is informed by Design for Manufacture rules based on powder bed fusion process capability. • The design tool is applied to five distinct curves and validated via manufacturing and testing of optimised designs. • The unit cell aspect ratio and strut taper design variables improve strength and energy absorption in BCC lattices. • Strong repeatability and ductility suitable for energy absoption applications is observed in as-built 17-4PH lattices. Additive manufacturing (AM) allows for the creation of novel complex structures previously impossible using traditional subtractive methods; however, there is a need for new design approaches to fully exploit this potential. Metal lattice structures have attracted attention for their broad customisable range of configurations and potential properties, and to take advantage of these possibilities there is a need for intelligent design tools to optimise the lattice according to the desired application and properties. This study presents and demonstrates a method to automatically design lattice structures which achieve a desired compressive stress–strain curve and satisfy user-defined manufacturing limits. A genetic algorithm iterates selected design variables (unit cell aspect ratio, strut taper, and thickness gradient) and minimises the error between the target stress–strain curve and predicted curve which is determined via automated finite element (FE) modelling of each lattice design. The optimised design is validated through manufacture and compression testing of 17-4PH stainless steel lattice structures, and micro-CT imaging to assess build quality. The tool's versatility is demonstrated by successfully designing lattices to fit a range of target curves. This work demonstrates the potential of this smart inverse design tool to exploit AM and optimise lattice design based on desired performance. The custom written design tool presented in this study is available open source at https://github.com/I-Form/LiDOT. [ABSTRACT FROM AUTHOR]

Streptococcus thermophilus strain ST64987 was exposed to a member of a recently discovered group of S. thermophilus phages (the 987 phage group), generating phage‐insensitive mutants, which were then characterized phenotypically and genomically. Decreased phage adsorption was observed in selected bacteriophage‐insensitive mutants, and was partnered with a sedimenting phenotype and increased cell chain length or aggregation. Whole genome sequencing of several bacteriophage‐insensitive mutants identified mutations located in a gene cluster presumed to be responsible for cell wall polysaccharide production in this strain. Analysis of cell surface‐associated glycans by methylation and NMR spectroscopy revealed a complex branched rhamno‐polysaccharide in both ST64987 and phage‐insensitive mutant BIM3. In addition, a second cell wall‐associated polysaccharide of ST64987, composed of hexasaccharide branched repeating units containing galactose and glucose, was absent in the cell wall of mutant BIM3. Genetic complementation of three phage‐resistant mutants was shown to restore the carbohydrate and phage resistance profiles of the wild‐type strain, establishing the role of this gene cluster in cell wall polysaccharide production and phage adsorption and, thus, infection. [ABSTRACT FROM AUTHOR]

The dairy fermentation industry relies on the activity of lactic acid bacteria in robust starter cultures to accomplish milk acidification. Maintenance of the composition of these starter cultures, whether defined or undefined, is essential to ensure consistent and high-quality fermentation end products. To date, limited information exists regarding the microbial composition of undefined starter culture systems. Here, we describe a culture-based analysis combined with a metagenomics approach to evaluate the composition of two undefined mesophilic starter cultures. In addition, we describe a qPCR-based genotype detection assay, which is capable of discerning nine distinct lactococcal genotypes to characterize these undefined starter cultures, and which can be applied to monitor compositional changes in an undefined starter culture during a fermentation. IMPORTANCE This study reports on the development of a combined culture-based analysis and metagenomics approach to evaluate the composition of two undefined mesophilic starter cultures. In addition, a novel qPCR-based genotype detection assay, capable of discerning nine distinct lactococcal genotypes (based on lactococcal cell wall polysaccharide biosynthesis gene clusters), was used to monitor compositional changes in an undefined starter culture following phage attack. These analytical approaches facilitate a multifaceted assessment of starter culture compositional stability during milk fermentation, which has become an important QC aspect due to the increasing demand for consistent and high-quality dairy products. [ABSTRACT FROM AUTHOR]

Background. Standardized tests and outcome measures (STOM) have not been consistently implemented as part of most physical therapists' practice. Incidence of STOM use among physical therapists at Beth Israel Deaconess Medical Center was similar to low levels cited nationally among acute care physical therapists. Targeted knowledge translation (KT) strategies have been suggested to promote the application of research evidence into clinical decision making. Purpose. The purpose of this quality improvement (QI) effort was to implement a series of interventions aimed at increasing both use and interpretation of STOM by physical therapists practicing in acute care. Design. This study used an observational longitudinal design. Methods. A literature review identified current barriers and facilitators to the use of STOM by physical therapists. KT strategies were tailored to the practice setting in order to target barriers and promote facilitators to the use of STOM. Data were collected through retrospective chart review at baseline and then subsequently at 4 periods following the implementation of the QI project. Results. A statistically significant increase in both the use (primary outcome) and interpretation (secondary outcome) of STOM was observed following the implementation of KT strategies. The increase was sustained at all subsequent measurement periods. Limitations. Limitations include the lack of a control group and the small number of setting- and diagnosis-specific STOM available for use by physical therapists practicing in acute care. Conclusions. Implementation of KT strategies was associated with an increase in the frequency of use and interpretation of STOM. Similar QI efforts are feasible in any acute care physical therapy department and potentially other settings. [ABSTRACT FROM AUTHOR]

Predation of starter lactic acid bacteria such as Streptococcus thermophilus by bacteriophages is a persistent and costly problem in the dairy industry. CRISPRmediated bacteriophage insensitive mutants (BIMs), while straightforward to generate and verify, can quickly be overcome by mutant phages. The aim of this study was to develop a tool allowing the generation of derivatives of commercial S. thermophilus strains which are resistant to phage attack through a non-CRISPR-mediated mechanism, with the objective of generating BIMs exhibiting stable resistance against a range of isolated lytic S. thermophilus phages. To achieve this, standard BIM generation was complemented by the use of the wild-type (WT) strain which had been transformed with an antisense mRNA-generating plasmid (targeting a crucial CRISPR-associated [cas] gene) in order to facilitate the generation of non-CRISPR-mediated BIMs. Phage sensitivity assays suggest that non-CRISPR-mediated BIMs exhibit some advantages compared to CRISPRmediated BIMs derived from the same strain. [ABSTRACT FROM AUTHOR]

We present the complete genome sequences of four members of a novel group of phages infecting Streptococcus thermophilus, designated here as the 987 group. Members of this phage group appear to have resulted from genetic exchange events, as evidenced by their "hybrid" genomic architecture, exhibiting DNA sequence relatedness to the morphogenesis modules of certain P335 group Lactococcus lactis phages and to the replication modules of S. thermophilus phages. All four identified members of the 987 phage group were shown to elicit adsorption affinity to both their cognate S. thermophilus hosts and a particular L. lactis starter strain. The receptor binding protein of one of these phages (as a representative of this novel group) was defined using an adsorption inhibition assay. The emergence of a novel phage group infecting S. thermophilus highlights the continuous need for phage monitoring and development of new phage control measures. [ABSTRACT FROM AUTHOR]

The author presents an analysis of postwar representation of Hollywood films which part or all of their plots set in New Zealand. He brings attention that Hollywood's vision of New Zealand cinema after World War II as stereotypes and untold true stories mixed in complicated ways. He stresses that Hollywood narratives dominated their consumption of film stories because the cinema images of even New Zealand in those decades were constructed according to the conventions and signifying systems of Hollywood. He attempts to analyze several Hollywood films shoot in New Zealand.

Highlights the modeling and control of a pneumatic robot which can be used for position tracking and end-effector force control. Information on previous methods used for controlled motion; Description of the robot utilized in this research; Examination of a pneumatic actuator model; Statistical data compiled in this research.

Summary: Available 3D structures of bacteriophage modules combined with predictive bioinformatic algorithms enabled the identification of adhesion modules in 57 siphophages infecting Streptococcus thermophilus (St). We identified several carbohydrate‐binding modules (CBMs) in so‐called evolved distal tail (Dit) and tail‐associated lysozyme (Tal) proteins of St phage baseplates. We examined the open reading frame (ORF) downstream of the Tal‐encoding ORF and uncovered the presence of a putative p2‐like receptor‐binding protein (RBP). A 21 Å resolution electron microscopy structure of the baseplate of cos‐phage STP1 revealed the presence of six elongated electron densities, surrounding the core of the baseplate, that harbour the p2‐like RBPs at their tip. To verify the functionality of these modules, we expressed GFP‐ or mCherry‐coupled Tal and putative RBP CBMs and observed by fluorescence microscopy that both modules bind to their corresponding St host, the putative RBP CBM with higher affinity than the Tal‐associated one. The large number of CBM functional domains in St phages suggests that they play a contributory role in the infection process, a feature that we previously described in lactococcal phages and beyond, possibly representing a universal feature of the siphophage host‐recognition apparatus. [ABSTRACT FROM AUTHOR]

Clinical empathy has been studied in a number of health-care disciplines suggesting that higher practitioner empathy leads to improved patient health and wellness and improved patient outcomes. While some aspects of the physical therapist-patient relationship have been described, evidence of quantitative assessment of clinical empathy in physical therapists is scarce. To investigate the level of self-reported clinical empathy in physical therapists and its relationship to practice environment and workplace engagement, the Jefferson Scale of Empathy-Health Provider version (JSE-HP) and the Oldenburg Burnout Inventory (OLBI) were used. Study participants were 123 physical therapists working full time at either an acute care setting, a rehabilitation hospital, or an outpatient clinic. These physical therapists demonstrated a mean JSE-HP score of 118.5 (9.1) and a mean OLBI score of 15.63 (3.5). This mean empathy score was found to be higher than reported empathy level of some health disciplines such as nursing and pharmacy yet lower than others such as mental health workers, psychiatrists, and pediatricians. Practice setting was not found to be a significant factor regarding empathy levels in physical therapists. As reported in previous studies, there was a positive correlation between being female and having higher empathy levels. A positive correlation was found between age and work disengagement. Finally, our hypothesis regarding a negative correlation between empathy and work disengagement was confirmed, suggesting that workplace disengagement may diminish a physical therapist's empathy, which may then negatively affect patient clinical outcomes. [ABSTRACT FROM AUTHOR]

Bifidobacterium breve is a common and sometimes very abundant inhabitant of the human gut. Genome sequencing of B. breve JCM 7017 revealed the presence of an extrachromosomal element, designated pMP7017 consisting of>190 kb, thus representing the first reported bifidobacterial megaplasmid. In silico characterization of this element revealed several genomic features supporting a stable establishment of the megaplasmid in its host, illustrated by predicted CRISPR-Cas functions that are known to protect the host against intrusion of foreign DNA. Interestingly, pMP7017 is also predicted to encode a conjugative DNA transfer apparatus and, consistent with this notion, we demonstrate here the conjugal transfer of pMP7017 to representative strains of B. breve and B. longum subsp. longum. We also demonstrate the presence of a megaplasmid with homology to pMP7017 in three B. longum subsp. longum strains. [ABSTRACT FROM AUTHOR]