Development of a monoclonal antibody specific to burbot (Lota lota) IgM and optimization of an ELISA to measure anti-Aeromonas sp. antibody titers following pathogen challenge.

Burbot (Lota lota) are an ideal candidate for cool or cold-water aquaculture and are gaining interest because of their high economic value, low temperature requirements, and fast growth rate. Limited information exists on the innate and adaptive immune systems of this species. This is partly due to the lack of species-specific tools to determine antibody responses following disease or vaccination or to characterize the immune response in general. An anti-IgM monoclonal antibody (mAb 27C) was developed and characterized via enzyme-linked immunosorbent assay (ELISA) and Western blot for species specificity, affinity to the heavy chain of burbot IgM, and cross-reactivity to other reagents used in the analysis. The 27C monoclonal antibody was further utilized to develop an ELISA protocol to measure the specific antibody response of burbot following exposure to two pathogenic strains of Aeromonas sp. (A141 and IR004). This ELISA confirmed that vaccinated burbot that survived the challenge with either strain developed statistically higher titers of anti-Aeromonas antibodies specific for the relative strain when compared to fish that were not vaccinated or challenged. Western blot analysis further demonstrated that burbot surviving challenge had serum IgM that recognized distinct antigens specific to the strain they were challenged with, A141 bound to antigens in the 50-250Kda range and IR004 bound to a distinct 150Kda antigen. Western blots further indicated that each strain shared antigenic regions regardless of experimental Aeromonas strain exposure. Finally, immunofluorescent staining confirmed that mAb 27C binds to membrane-bound IgM (presumably B cells) on burbot head kidney cells. Taken together, results from this study demonstrate that mAb 27C specifically recognized burbot IgM and will be an important tool to further characterize the adaptive and cellular immune responses of this fish species.

Characterization of maternal immunity following vaccination of broodstock against IHNV or Flavobacterium psychrophilum in rainbow trout (Oncorhynchusmykiss).

Infectious hematopoietic necrosis (IHN) is a significant viral disease affecting salmonids, whereas Flavobacterium psychrophilum (Fp), the causative agent of bacterial coldwater disease (BCWD), remains one of the most significant bacterial pathogens of salmonids. We explored maternal immunity in the context of IHN and BCWD management in rainbow trout (Oncorhynchus mykiss) aquaculture. Two experimental trials were conducted where different groups of female broodstock were immunized prior to spawning with an IHNV DNA vaccine or a live attenuated F. psychrophilum (Fp B.17-ILM) vaccine alone, or in combination. Progeny were challenged with either a low or high dose of IHNV at 13 days post hatch (dph) and 32 dph or challenged with F. psychrophilum at 13 dph. Mortality following a low-dose IHNV challenge at 13 dph was significantly lower in progeny from vaccinated broodstock vs. unvaccinated broodstock, but no significant differences were observed at 32 dph. Mortality due to BCWD was also significantly reduced in 13 dph fry that originated from broodstock immunized with the Fp B.17-ILM vaccine. After vaccination broodstock developed specific or neutralizing antibodies respectively to F. psychrophilum and IHNV; however, antibody titers in eggs and fry were undetectable. In the eggs and fry mRNA transcripts of the complement components C3 and C5 were detected at much higher levels in progeny from vaccinated broodstock and showed a significantly increased and rapid response post-challenge compared with unvaccinated broodstock. After challenges pro-inflammatory cytokine expression was immediately and considerably elevated in the fry from vaccinated broodstock vs. unvaccinated broodstock, whereas adaptive immune genes were elevated to a lesser degree. Results suggest that maternal transfer of innate and adaptive factors at the transcript level occurred because development of lymphomyeloid organs is not complete in such young fry. In addition to documenting maternally derived immunity in teleosts, this study demonstrates that broodstock vaccination can confer some degree of protection to progeny against viral and bacterial pathogens.

Mapping the cis-regulatory architecture of the human retina reveals noncoding genetic variation in disease.

The interplay of transcription factors and cis-regulatory elements (CREs) orchestrates the dynamic and diverse genetic programs that assemble the human central nervous system (CNS) during development and maintain its function throughout life. Genetic variation within CREs plays a central role in phenotypic variation in complex traits including the risk of developing disease. We took advantage of the retina, a well-characterized region of the CNS known to be affected by pathogenic variants in CREs, to establish a roadmap for characterizing regulatory variation in the human CNS. This comprehensive analysis of tissue-specific regulatory elements, transcription factor binding, and gene expression programs in three regions of the human visual system (retina, macula, and retinal pigment epithelium/choroid) reveals features of regulatory element evolution that shape tissue-specific gene expression programs and defines regulatory elements with the potential to contribute to Mendelian and complex disorders of human vision.

Rhinoplasty with Concurrent Repair of Septal Perforation.

Concurrently performing septal perforation repair and rhinoplasty is challenging. Successful management of patients who require or desire these procedures performed simultaneously can be achieved with appropriate preoperative workup and optimization, thoughtful surgical planning, masterful execution, and diligent postoperative care. Consideration of perforation etiology and status of the perforation along with overall nasal health are the foundation upon which surgery may be planned. Preoperative evaluation to determine locations of obstruction, aesthetic goals, and feasibility of procedure is required. Rhinoplasty techniques and perforation closure techniques may share common maneuvers making concurrent surgery possible. Surgical techniques to achieve perforation closure often influence rhinoplasty considerations, and vice versa. Care must be taken to account and possibly even to capitalize on these influences. Perforation closure techniques that may be readily combined with rhinoplasty include interposition grafting and intranasal mucosal rotational advancement flaps, which may exert forces upon the nose with aesthetic or functional consequences. Rhinoplasty techniques targeting any third of the nose also can impact tension or available mucosa for perforation closure techniques. Combining these procedures can be very rewarding for the patient and provider but should be considered responsibly and thoughtfully.

Production of a monoclonal antibody specific to sablefish (Anoplopoma fimbria) IgM and its application in ELISA, western blotting, and immunofluorescent staining.

Sablefish (Anoplopoma fimbria) are an emerging aquaculture species native to the continental shelf of the northern Pacific Ocean. There is limited information on both innate and adaptive immunity for this species and new tools are needed to determine antibody response following vaccination or disease outbreaks. In this paper, a monoclonal antibody, UI-25A, specific to sablefish IgM was produced in mice. Western blotting confirmed UI-25A recognizes the heavy chain of IgM and does not cross react to proteins or carbohydrates in serum of four other teleost species. An ELISA was developed to measure Aeromonas salmonicida specific IgM in the plasma of sablefish from a previous experiment where fish were immunized with a proprietary A. salmonicida vaccine. UI-25A was used in Western blot analyses to identify immunogenic regions of A. salmonicida recognized by this specific IgM from vaccinated sablefish. Immunofluorescent staining also demonstrated the ability of UI-25A to recognize membrane-bound IgM and identify IgM + cells in the head kidney. These results demonstrate the usefulness of UI-25A as a tool to improve the understanding of antibody-mediated immunity in sablefish as well as to provide valuable information for vaccine development and expansion of aquaculture efforts for this fish species.

Calculations of pKa Values for a Series of Naturally Occurring Modified Nucleobases.

Modified nucleobases are found in functionally important regions of RNA and are often responsible for essential structural roles. Many of these nucleobase modifications are dynamically regulated in nature, with each modification having a different biological role in RNA. Despite the high abundance of modifications, many of their characteristics are still poorly understood. One important property of a nucleobase is its pKa value, which has been widely studied for unmodified nucleobases, but not for the modified versions. In this study, the pKa values of modified nucleobases were determined by performing ab initio quantum mechanical calculations using a B3LYP density functional with the 6-31+G(d,p) basis set and a combination of implicit-explicit solvation systems. This method, which was previously employed to determine the pKa values of unmodified nucleobases, is applicable to a variety of modified nucleobases. Comparisons of the pKa values of modified nucleobases give insight into their structural and energetic impacts within nucleic acids.

Minimally Invasive Nasal Valve Treatment.

The objective of this article is to explore minimally invasive techniques to treat a poorly functioning nasal valve. It reviews the literature to introduce anatomy and examination, detail efficacy, instruct procedural protocols, and summarize utility. The article was composed based upon literature review and expert opinion. Nasal obstruction is a frequent problem managed by the facial plastic surgeon. It can occur at multiple sites, but commonly problematic regions are the nasal valves. A comprehensive nasal exam can elucidate the source of obstruction and validated obstruction measurements, such as the Nasal Obstruction Symptom Evaluation Instrument or the Standardized Cosmesis and Health Nasal Outcomes Survey for Functional and Cosmetic Rhinoplasty, can classify severity. Cartilage grafting and open septorhinoplasty techniques are invaluable for correcting nasal valve obstruction; however, the traditional methods may be neither possible nor optimal for all patients. Minimally invasive treatment modalities have emerged to offer an alternative to classical treatment algorithms. Minimally invasive techniques aimed at improving the nasal valve function include nasal dilators, suture suspension, lateral nasal wall bioresorbable implants, and cartilaginous radio frequency remodeling. Benefits of these methods include clinic-based intervention, avoidance of general anesthesia, use in previously operated noses, and minimally invasive nature. Publications with ranging levels of evidence support the efficacy of these interventions when patients with isolated nasal valve obstruction are examined. Limitations in the data, such as study power, source of funding, duration of follow-up, and longevity of response, warrant the need for further investigation; however, the efficacy of these techniques justifies implementation in proper patient populations into current practice. Minimally invasive nasal valve correction is a valuable tool in the armamentarium of the facial plastic surgeon. It cannot and should not replace traditional septorhinoplasty; however, the techniques are a valuable consideration for the appropriate patient population.

Assessment of Flavobacterium psychrophilum-associated mortality in Atlantic salmon (Salmo salar) and brook trout (Salvelinus fontinalis).

Salmonid diseases caused by infections of Flavobacterium psychrophilum, the causative agent of bacterial coldwater disease, remain difficult to manage as novel, pathogenic strains continue to emerge in aquaculture settings globally. To date, much of the research regarding treatment options and vaccine development has focused on rainbow trout (Oncorhynchus mykiss), but other inland-reared salmonids are also impacted by this Gram-negative bacterium. As such, Atlantic salmon (Salmo salar) and brook trout (Salvelinus fontinalis) were injection-challenged with a variety of previously reported F. psychrophilum strains isolated from disease diagnostic cases in salmonids, as well as a standard and well-studied F. psychrophilum strain (CSF 259-93) known to be virulent in rainbow trout. In three separate virulence assessments (Trials A, B and C), strains US063 (isolated from lake trout; Salvelinus namaycush) and US149 (isolated from Atlantic salmon) caused a significantly higher cumulative per cent mortality (CPM) relative to other strains in Atlantic salmon (p <.001 for all trials), with US149 causing significantly greater mortality than US063 in Trials A (CPM 97% vs. 65%, p =.008) and B (CPM 96% ± 2.3% vs. 81.33% ± 4.8%, p =.014). Trial C used a lower dose (1.86 × 108  CFU/mL) for US149, resulting in a lower mortality (78.67% ± 9.33%) relative to Trials A and B. CSF259-93 did not cause significant mortality in any trials. In brook trout, the strain 03-179 (originally isolated from steelhead trout; Oncorhynchus mykiss) was significantly more virulent than any other (CPM 100% ± 0%, p <.001), followed by US063 (73% ± 3.8%) and US149 (40% ± 6.1%,) respectively. Again, CSF259-93 did not cause significant mortality relative to a mock challenge treatment. Results provide information about the applicability of strain selection in F. psychrophilum virulence testing in Atlantic salmon and brook trout, demonstrating the high virulence of US063 and US149 for these salmonid species. This information is applicable for the development of therapeutics and vaccines against F. psychrophilum infections and demonstrates the reproducibility of the experimental challenge model.

Isolation and experimental challenge of cultured burbot (Lota lota maculosa) with Flavobacterium columnare and Aeromonas sp. isolates.

Burbot (Lota lota maculosa) are a potential new species for commercial aquaculture. As burbot culture expands, there is a need to further define pathogen susceptibility and characterize aspects of the burbot immune response in an effort to assess fish health. A recent clinical diagnostic case from juvenile burbot reared at a commercial production facility resulted in the isolation and identification of Flavobacterium columnare along with several Aeromonas spp. The F. columnare isolate was assigned to genetic group 1 via multiplex PCR, a genetic group commonly associated with columnaris disease cases in rainbow trout (Oncorhynchus mykiss). Virulence of the F. columnare isolate was assessed in vivo in both juvenile burbot and rainbow trout. Additionally, several of the Aeromonas sp. case isolates were identified via sequencing (16S rRNA, gyrB and rpoD) and a putative A. sobria isolate (BI-3) was used to challenge burbot, along with a known virulent Aeromonas sp. (A141), but BI-3 was not found to be virulent. Burbot were refractory to F. columnare when challenged by immersion, and it is likely that this is a secondary pathogen for burbot. Although refractory in burbot, the identified F. columnare isolate (BI-1) was found to be virulent in rainbow trout.

Lithography-Free Electrochemical Patterning of Conductive Substrates with Metal Oxides.

Reactive interface patterning promoted by lithographic electrochemistry serves as a facile method for generating submicron structures on conductive substrates. A binary-potential step applied to a metal layer with a resist overlayer allows silicon to be patterned with metal oxides. In this study, the role and influence of the resist overlayer on the uniformity of pattern formation are examined. The ability of the resist to detach from the underlying metal is a critical determinant of pattern geometry. By choosing an appropriate resist, large patterns with submicron precision are generated quickly by the application of the binary-potential steps. From this information, a lithography-free approach to generating identical patterns is achieved with simple resists such as that furnished from a lacquer-water emulsion, thus greatly simplifying the patterning of silicon with metal oxide catalysts.

Targeting Suppressive Oligonucleotide to Lymph Nodes Inhibits Toll-like Receptor-9-Mediated Activation of Adaptive Immunity.

PURPOSE: This paper aims to investigate the immunoinhibitory properties of a lymph nodes-targeting suppressive oligonucleotide (ODN) for the potential treatment of autoimmune diseases or chronic inflammation. METHODS: Synthetic suppressive ODN engineered with an albumin-binding diacyl lipid at the 5'-terminal (lipo-ODN) was synthesized. In vitro and in vivo experiments were designed to compare the immune suppressive properties of lipo-ODN and unmodified ODN. Cellular uptake and distribution, inhibition of Toll-like receptor (TLR) activation, lymph nodes (LN) draining, and the suppression of antigen-specific immune responses in an ovalbumin protein model was investigated. RESULTS: Compared to unmodified ODN, lipid functionalized suppressive ODN demonstrated enhanced cellular uptake and TLR-9 specific immune suppression in TLR reporter cells. Additionally, injection of a low dose of lipid-modified suppressive ODN, but not the unconjugated ODN, accumulated in the draining LNs and exhibited potent inhibition of antigen-specific CD8+ T cell and B cell responses in vivo. CONCLUSIONS: Targeting suppressive ODN to antigen presenting cells (APCs) in the local LNs is an effective approach to amplify the immune modulation mediated by ODN containing repetitive TTAGGG motif. This approach might be broadly applicable to target molecular adjuvants to the key immune cells in the LNs draining from disease site, providing a simple strategy to improve the efficacy of many molecular immune modulators.

Mortality and morbidity of neck of femur fractures: A comparison between Orange and peripheral centres.

OBJECTIVE: The aim of this study is to examine and compare the mortality and morbidity of patients with neck of femur (NOF) fracture who presented initially to either Orange Health Service (OHS), a secondary orthopaedic referral centre or to a peripheral hospital and subsequently transferred to OHS. DESIGN, SETTING AND PARTICIPANTS: This study is designed as a retrospective review of medical records of 151 patients with NOF fractures who were treated operatively between March 2013 and March 2014 at OHS. Patients were divided into OHS group and 'Other' group based on their initial presenting location. MAIN OUTCOME MEASURES: Mortality, unplanned intensive care unit/high dependency unit admissions, ischemic heart/cerebral events, infection rates and length of stay. RESULTS: There was no statistically significant difference in mortality and morbidity proportions between both groups. There were significant differences between lengths of stay in the acute setting which was shorter in the 'Other' group. CONCLUSION: Contrary to the literature, this study found that the morbidity and mortality outcomes of patients with NOF fracture who presented initially to rural hospitals were equivalent to those who presented to a secondary orthopaedic referral centre. However, those who first presented to rural hospitals had a shorter duration of stay in the acute setting.

Facile, rapid, and large-area periodic patterning of semiconductor substrates with submicron inorganic structures.

The development of high-throughput and scalable techniques for patterning inorganic structures is useful for the improved function and efficiency of photonic and energy conversion devices. Here we demonstrate a facile and rapid electrochemical method for patterning periodic metallic and nonmetallic submicron structures over large areas. Si substrates have been patterned with arrays of periodically spaced lines, rings, squares, and terraces of main-group and transition-metal oxides. In addition to planar substrates, three-dimensional surfaces and their vertical sidewalls have been patterned. The features are 20(±1) nm high and 360(±15) nm wide, and their period is finely tunable in situ from 500 nm to 7 µm. These features exhibit <3% variation in period and are rapidly patterned in <2 min. We demonstrate the versatility of the technique by rapidly patterning an efficient water splitting catalyst, Co phosphate oxide (CoPi), and show that the integrated materials system performs water splitting with complete Faradaic efficiency. More generally, the ability to pattern submicron structures over large areas in a facile, reliable, and timely manner may be useful for the fabrication of devices for energy, meta-material, and sensing applications.

Are we developing walkable suburbs through urban planning policy? Identifying the mix of design requirements to optimise walking outcomes from the 'Liveable Neighbourhoods' planning policy in Perth, Western Australia.

BACKGROUND: Planning policy makers and practitioners are requesting clearer guidance on the 'essential' ingredients as assessed by public health researchers to ensure suburban neighbourhood environments are designed to promote active living behaviours such as walking. OBJECTIVES: To identify the combination of design requirements from the 'Liveable Neighbourhoods' (LN) planning policy in Perth, Western Australia that were optimally supportive of walking. METHODS: K-means cluster analysis identified groups of developments with homogeneous LN features from its community design (CD), movement network (MN), lot layout (LL) and public parkland (PP) elements. Walking behaviours measured using the Neighbourhood Physical Activity Questionnaire were compared between participants resident in the different clusters, adjusting for demographic characteristics, self-selection factors, stage of construction and scale of development. RESULTS: Compared with participants living in the referent cluster of 'poor CD and PP developments' those living in: 'MN and LL developments' had higher odds of doing any (OR = 1.74; 95 % CI = 1.22, 2.48) and ≥60 min walking for recreation (WR) (OR = 2.05; 1.46, 2.88); 'PP developments' had increased odds of doing any WR (OR = 3.53; 2.02, 6.17), ≥60 min WR (OR = 3.37; 1.98, 5.74) and any total walking (TW) (OR = 2.35; 1.36, 4.09); 'CD-MN developments' had increased odds of doing any walking for transport (WT) (OR = 2.64; 1.38, 5.06), ≥60 min WT (OR = 1.98; 1.09, 3.61), any TW (OR = 1.71; 1.44, 2.03), ≥60 min TW (OR = 1.77; 1.14, 2.76) and ≥150 min TW (OR = 1.47; 1.15, 1.86). CONCLUSIONS: This study is the first to have empirically identified a mix of specific and distinguishing planning policy neighbourhood design requirements to optimise walking outcomes. These findings will assist in the assessment of urban plans for greenfield suburban developments designed to promote walking and physical activity.

How preparation to touch or grasp alters visual size perception.

Prior studies have suggested that visually guided actions are resistant to the effects of some pictorial size illusions, e.g., the maximum grip aperture component of a grasp for an element of the Ebbinghaus illusion display. We present evidence that when participants prepare to grasp, the reduction in illusion magnitude observed for action components is also present for conscious perceptual judgments. Our studies characterize how visual size perception changes when we choose to engage in different size-mediated behaviors. Even when the stimuli used were identical for two different tasks, we found that available information was processed differently. In the studies, participants always selected which of the two targets was larger. In some conditions, the context in which the targets were presented induced a visual illusion of size. We varied the sizes of target pairs to assess the magnitude of these visual illusions. In some tasks, participants indicated their size choice verbally. For other tasks, participants reached to grasp or touch the target that they perceived as larger. Illusion magnitudes were smaller when participants engaged in actions directed at a target or when participants imagined performing those actions. This shift in visual processing persisted for several minutes after participants switched back to a verbal, non-grasping, non-touch task. A motor interference task eliminated the reduction in illusion magnitude.

Automatic sleep spindles identification and classification with multitapers and convolution.

Sleep spindles are isolated transient surges of oscillatory neural activity present during sleep stages 2 and 3 in the nonrapid eye movement (NREM). They can indicate the mechanisms of memory consolidation and plasticity in the brain. Spindles can be identified across cortical areas and classified as either slow or fast. There are spindle transients across different frequencies and power, yet most of their functions remain a mystery. Using several electroencephalogram (EEG) databases, this study presents a new method, called the "spindles across multiple channels" (SAMC) method, for identifying and categorizing sleep spindles in EEGs during the NREM sleep. The SAMC method uses a multitapers and convolution (MT&C) approach to extract the spectral estimation of different frequencies present in sleep EEGs and graphically identify spindles across multiple channels. The characteristics of spindles, such as duration, power, and event areas, are also extracted by the SAMC method. Comparison with other state-of-the-art spindle identification methods demonstrated the superiority of the proposed method with an agreement rate, average positive predictive value, and sensitivity of over 90% for spindle classification across the three databases used in this paper. The computing cost was found to be, on average, 0.004 seconds per epoch. The proposed method can potentially improve the understanding of the behavior of spindles across the scalp and accurately identify and categories sleep spindles.

The potential of repetitive transcranial magnetic stimulation for addressing sleep difficulties in children with autism - A brief communication.

Sleep difficulties can co-occur with autistic traits and have been frequently reported in children diagnosed with autism. Thus, sleep difficulties may impact neural development, cognition, and behavioural functioning in children with autism. Interventions, such as repetitive transcranial magnetic stimulation (rTMS), that target aberrant neural structures underpinning autistic traits and sleep difficulties in children could have beneficial effects. The rTMS effects on the pathophysiological pathways hypothesised to underpin autism and sleep difficulties are well-established in the literature; however, clinical evidence of its potential to improve sleep difficulties in children with autism is limited. While the preliminary data is promising, further robust rTMS studies are warranted to encourage its use in clinical practices.

Systematic Review and Meta-Analysis of Endovascular Therapy Effectiveness for Unruptured Saccular Intracranial Aneurysms.

Background: Currently, endovascular treatment of intracranial aneurysms (ICAs) is limited by low complete occlusion rates. The advent of novel endovascular technology has expanded the applicability of endovascular therapy; however, the superiority of novel embolic devices over the traditional Guglielmi detachable coils (GDCs) is still debated. We performed a systematic review of literature that reported Raymond-Roy occlusion classification (RROC) rates of modern endovascular devices to determine their immediate and follow-up occlusion effectiveness for the treatment of unruptured saccular ICAs. Methods: A search was conducted using electronic databases (PUBMED, Cochrane, ClinicalTrials.gov, Web of Science). We retrieved studies published between 2000-2022 reporting immediate and follow-up RROC rates of subjects treated with different endovascular ICA therapies. We extracted demographic information of the treated patients and their reported angiographic RROC rates. Results: A total of 80 studies from 15 countries were included for data extraction. RROC rates determined from angiogram were obtained for 21,331 patients (72.5% females, pooled mean age: 58.2 (95% CI: 56.8-59.6), harboring 22,791 aneurysms. The most frequent aneurysm locations were the internal carotid artery (46.4%, 95% CI: 41.9%-50.9%), the anterior communicating artery (26.4%, 95% CI: 22.5%-30.8%), the middle cerebral artery (24.5%, 95% CI:19.2%-30.8%) and the basilar tip (14.4%, 95% CI:11.3%-18.3%). The complete occlusion probability (RROC-I) was analyzed for GDCs, the Woven EndoBridge (WEB), and flow diverters. The RROC-I rate was the highest in balloon-assisted coiling (73.9%, 95% CI: 65.0%-81.2%) and the lowest in the WEB (27.8%, 95% CI:13.2%-49.2%). The follow-up RROC-I probability was homogenous in all analyzed devices. Conclusions: We observed that the coil-based endovascular therapy provides acceptable rates of complete occlusion, and these rates are improved in balloon-assisted coils. Out of the analyzed devices, the WEB exhibited the shortest time to achieve >90% probability of follow-up complete occlusion (~18 months). Overall, the GDCs remain the gold standard for endovascular treatment of unruptured saccular aneurysms.

Chiropractors in Multidisciplinary Teams: Enablers of Colocation Integration in GP-Led Primary Healthcare.

The aim of this study was to explore and document the enablers and barriers of chiropractic care colocation in general practice at a large-scale private primary care centre in Australia. This study focused on the perceptions of healthcare professionals regarding this integration. The research setting was a large integrated primary care centre located in an outer metro, low-socioeconomic area in the City of Moreton Bay, Queensland, Australia. Participant inclusion criteria included general medical practitioners, practice nurses, and medical managers who self-reported interactions with the physically collocated and integrated chiropractic practice. Data was collected from 22 participants using face-to-face, qualitative, semi-structured interviews with an average duration of 32 min. The data collected included perceptions of chiropractic treatment, enablers to patient referral pathways, and views of the integrated chiropractic care model. A reflexive thematic analysis was conducted on the data set. All participants reported that this was their first exposure to the colocation of a chiropractor within a general medical practice. Four key enablers of chiropractic care integration were identified: (1) the practitioner [chiropractor], (2) the organisation [general practice], (3) consumer flow, and (4) the environment [shared spaces and tenant ecosystem]. The chiropractic integration enhanced knowledge sharing and interprofessional trust among healthcare providers. The formal reporting of patient outcomes and understanding of the chiropractor's scope of practice further enabled referrals to the service. Shared administrative and business processes, including patient records, booking systems, and clinical meetings, facilitated relationship development between the chiropractor and referring health providers. Colocation as part of a larger primary care centre created proximity and convenience for health providers in terms of interprofessional communication, and for patients, in terms of access to chiropractic services. Existing governance structures supported communication, professional education, and shared values related to the delivery of patient-centred care. Identified barriers included limited public funding for chiropractic services resulting in reduced access for patients of low-socioeconomic status. Additionally, scepticism or negativity towards the discipline of chiropractic care was identified as an initial barrier to refer patients. In most cases, this view towards the chiropractor was overcome by regular patient reporting of positive treatment outcomes to their GP, the delivery of education sessions by the chiropractor for the health providers, and the development of interprofessional trust between the chiropractor and referring health providers. This study provides preliminary evidence and a conceptual framework of factors influencing the successful integration of chiropractic care within an Australian large primary care centre. The data collected indicated that integration of chiropractic care into a primary care centre serving a low-socioeconomic region can be achieved with a high degree of health provider satisfaction.

Rapid IDH1-R132 genotyping panel utilizing locked nucleic acid loop-mediated isothermal amplification.

While the detection of single-nucleotide variants (SNVs) is important for evaluating human health and disease, most genotyping methods require a nucleic acid extraction step and lengthy analytical times. Here, we present a protocol which utilizes the integration of locked nucleic acids (LNAs) into self-annealing loop primers for the allelic discrimination of five isocitrate dehydrogenase 1 R132 (IDH1-R132) variants using loop-mediated isothermal amplification (LAMP). This genotyping panel was initially evaluated using purified synthetic DNA to show proof of specific SNV discrimination. Additional evaluation using glioma tumor lysates with known IDH1-R132 mutational status demonstrated specificity in approximately 35 min without the need for a nucleic acid extraction purification step. This LNA-LAMP-based genotyping assay can detect single base differences in purified nucleic acids or tissue homogenates, including instances where the variant of interest is present in an excess of background wild-type DNA. The pH-based colorimetric indicator of LNA-LAMP facilitates convenient visual interpretation of reactions, and we demonstrate successful translation to an end-point format using absorbance ratio, allowing for an alternative and objective approach for differentiating between positive and negative reactions. Importantly, the LNA-LAMP genotyping panel is highly reproducible, with no false-positive or false-negative results observed.