Could it be monkeypox? Use of an AI-based epidemic early warning system to monitor rash and fever illness.

OBJECTIVES: The EPIWATCH artificial intelligence (AI) system scans open-source data using automated technology and can be used to detect early warnings of infectious disease outbreaks. In May 2022, a multicountry outbreak of Mpox in non-endemic countries was confirmed by the World Health Organization. This study aimed to identify signals of fever and rash-like illness using EPIWATCH and, if detected, determine if they represented potential Mpox outbreaks. STUDY DESIGN: The EPIWATCH AI system was used to detect global signals for syndromes of rash and fever that may have represented a missed diagnosis of Mpox from 1 month prior to the initial case confirmation in the United Kingdom (7 May 2022) to 2 months following. METHODS: Articles were extracted from EPIWATCH and underwent review. A descriptive epidemiologic analysis was conducted to identify reports pertaining to each rash-like illness, locations of each outbreak and report publication dates for the entries from 2022, with 2021 as a control surveillance period. RESULTS: Reports of rash-like illnesses in 2022 between 1 April and 11 July (n = 656 reports) were higher than in the same period in 2021 (n = 75 reports). The data showed an increase in reports from July 2021 to July 2022, and the Mann-Kendall trend test showed a significant upward trend (P = 0.015). The most frequently reported illness was hand-foot-and-mouth disease, and the country with the most reports was India. CONCLUSIONS: Vast open-source data can be parsed using AI in systems such as EPIWATCH to assist in the early detection of disease outbreaks and monitor global trends.

Horses are susceptible to natural, but resistant to experimental, infection with the liver fluke, Fasciola hepatica.

Fasciola hepatica is a common parasite of livestock in Ireland, causing significant economic losses and affecting animal welfare. A previous abattoir study of 200 horses led to an estimated 9.5 % prevalence of infection in horses slaughtered in Ireland. However, the epidemiology and pathogenic significance of this infection in this species is not well-described. The objectives of this study were to determine the susceptibility of horses to oral challenge infection with F. hepatica metacercariae, and to document the course of the infection along with serological and biochemical response. We attempted an experimental infection of horses (n = 10; 9 geldings and 1 mare) with F. hepatica. Four were given 1000 metacercariae, four 500 metacercariae and two were sham-infected. Blood and faecal samples were taken at intervals up to 18 weeks post-infection (wpi). ELISA assays were used to assess sero-conversion in the experimental horses and also in a panel of sera from horses of known fluke status. No flukes were recovered from any of the livers, and neither were any lesions that could be attributed to F. hepatica infection observed. Coproantigen ELISA was negative throughout for all horses. Three antibody detection ELISAs, useful in diagnosing fasciolosis in other species, had limitations as diagnostic aids as determined using a panel of sera from horses of known F. hepatica infection status. This study is limited by the relatively small number of animals included, and the relatively short duration of the study period. Failure to establish infection after oral challenge raises fundamental questions on the pathophysiology and epidemiology of equine fasciolosis.

Optimising the biocompatibility of 3D printed photopolymer constructs in vitro and in vivo.

3D printing is a rapid and accessible fabrication technology that engenders creative custom design solutions for cell scaffolds, perfusion systems and cell culture systems for tissue engineering. Critical to its success is the biocompatibility of the materials used, which should allow long-term tissue culture without affecting cell viability or inducing an inflammatory response for in vitro and in vivo applications. Polyjet 3D printers offer arguably the highest resolution with the fewest design constraints of any commercially available 3D printing systems. Although widely used for rapid-prototyping of medical devices and 3D anatomical modelling, polyjet printing has not been adopted by the tissue engineering field, largely due to the cytotoxicity of leachates from the printed parts. Biocompatibility in the context of cell culture is not commonly addressed for polyjet materials, as they tend to be optimised for their ability to fabricate complex structures. In order to study the potential issues surrounding the leaching of toxins, we prepared cell culture substrates using the commercially available MED610 photopolymer. The substrates were cleaned using either the manufacturer-specified 'biocompatible' washing procedures, or a novel protocol incorporating a sonication in isopropanol and water step. We then compared the effectiveness of these both in vitro and in vivo. Using primary mouse myoblast cultures, the manufacturer's protocol led to inconsistent and poorer cell viability when compared to the sonication protocol (p = 0.0002 at 48 h after indirect exposure). Subdermal implantation of MED610 into nude rats demonstrated a significant foreign body response with a greater number of giant cells (p = 0.0161) and foreign bodies (p = 0.0368) when compared to the sonication protocol, which was comparable to the control (sham) groups. These results present an improved, cytocompatible cleaning protocol of printable photopolymers to facilitate creative 3D-printed custom designs for cell culture systems for both in vitro and in vivo tissue engineering applications.

Prevalence of liver fluke infection in Irish horses and assessment of a serological test for diagnosis of equine fasciolosis.

BACKGROUND: There is little information on the prevalence of Fasciola hepatica infection in the horse population in Ireland or the potential impact of fluke infection on animal health. OBJECTIVES: To investigate F. hepatica infection in the Irish horse population and to assess the diagnostic potential of an indirect enzyme-linked immunosorbent assay (ELISA) based on the F. hepatica recombinant cathepsin L1 (CL1) antigen. STUDY DESIGN: Cross-sectional abattoir survey of horses for liver fluke status. METHODS: Animals (n = 200) were examined at an abattoir between May 2013 and April 2014. Horses were graded ante mortem for body condition score. Blood and faeces were collected and livers were examined post mortem by gross morphology. A cohort (n = 35) of livers were also examined histologically. Haematology and blood biochemistry, including serum liver enzyme activities, were measured and faeces were sedimented for egg counts. Serum was assayed by indirect ELISA using a recombinant CL1. RESULTS: The prevalence of liver fluke infection was 9.5%. There was no correlation between liver fluke status and time of year, breed classification, age group, sex, body condition score, ante mortem assessment, strongyle infection status, serum liver enzyme activities or CL1 concentration. A comparison of the CL1 ELISA in horse sera compared with a reference standard diagnosis showed high specificity of 95.6% (95% confidence interval [CI] 91.5-98.0%), but low sensitivity of 42.1% (95% CI 20.2-66.5%). MAIN LIMITATIONS: This study is limited by its nature as an abattoir study, the relatively small number of animals examined (n = 200), and the absence of a known negative group of horses. CONCLUSIONS: Blood biomarkers are not good indicators of liver fluke infection and the CL1 ELISA is not a sensitive tool for diagnosis of fluke infection in the horse. The prevalence of F. hepatica in horses indicates that further research is required to assess the potential impact of liver fluke on equine liver health.

Similar interaction chromatography of proteins: A cross interaction chromatographic approach to estimate the osmotic second virial coefficient.

Self-interaction chromatography (SIC) has established itself as an important experimental technique for the measurement of the second osmotic virial coefficients B22. B22 data are critical for understanding a range of protein solution phenomena, particularly aggregation and crystallisation. A key limitation to the more extensive use of SIC is the need to develop a method for immobilising each specific protein of interest onto a chromatographic support. This requirement is both a time and protein consuming constraint, which means that SIC cannot be used as a high throughput method for screening a wide range of proteins and their variants. Here an experimental framework is presented for estimating B22 values using Similar Interaction Chromatography (SimIC). This work uses experimental B23 and B32 data for lysozyme, lactoferrin, catalase and concanavalin A to reliably estimate B22 using arithmetic mean field approximations and is demonstrated to give good agreement with SIC measurements of B22 for the same proteins. SimIC could form the basis of a rapid protein variant screening methods to assess the developability of protein therapeutic candidates for industrial and academic researchers with respect to aggregation behaviour by eluting target proteins through a series of well-characterised protein immobilized reference columns.

Conductive composite fibres from reduced graphene oxide and polypyrrole nanoparticles.

Continuous composite fibres composed of polypyrrole (PPy) nanoparticles and reduced graphene oxide (rGO) at different mass ratios were fabricated using a single step wet-spinning approach. The electrical conductivity of the composite fibres increased significantly with the addition of rGO. The mechanical properties of the composite fibres also improved by the addition of rGO sheets compared to fibres containing only PPy. The ultimate tensile strength of the fibres increased with the proportion of rGO mass present. The elongation at break was greatest for the composite fibre containing equal mass ratios of PPy nanoparticles and rGO sheets. L929 fibroblasts seeded onto fibres showed no reduction in cell viability. To further assess toxicity, cells were exposed to media that had been used to extract any aqueous-soluble leachates from developed fibre. Overall, these composite fibres show promising mechanical and electrical properties while not significantly impeding cell growth, opening up a wide range of potential applications including nerve and muscle regeneration studies.

The second virial coefficient as a predictor of protein aggregation propensity: A self-interaction chromatography study.

The second osmotic virial coefficients (b2) of four proteins - lysozyme, recombinant human lactoferrin, concanavalin A and catalase were measured by self-interaction chromatography (SIC) in solutions of varying salt type, concentration and pH. Protein aggregate sizes based on the initial hydrodynamic radius of the protein solution species present were measured using dynamic light scattering, and the relationship between b2 and protein aggregate size was studied. A linear correlation was established between b2 values and protein aggregate hydrodynamic size for all proteins, and for almost all solution conditions. Aggregate sizes of <∼10nm, indicative of non-aggregated protein systems, were consistently observed to have b2 values >0. The observed b2 trends as a function of solution conditions were very much protein dependent, with notable trends including the existence of attractive interactions (negative b2 values) at low ionic strengths for catalase and concanavalin A, and the highly positive b2 values observed for lactoferrin over a wide range of solution conditions, reflecting lactoferrin's innately high stability. It is concluded that the quantification of protein-protein interactions using SIC based b2 data is a potentially valuable screening tool for predicting protein aggregation propensity.

The role of functional dopamine-transporter SPECT imaging in parkinsonian syndromes, part 1.

SUMMARY: As we defeat infectious diseases and cancer, one of the greatest medical challenges facing us in the mid-21st century will be the increasing prevalence of degenerative disease. Those diseases, which affect movement and cognition, can be the most debilitating. Dysfunction of the extrapyramidal system results in increasing motor disability often manifest as tremor, bradykinesia, and rigidity. The common pathologic pathway of these diseases, collectively described as parkinsonian syndromes, such as Parkinson disease, multiple system atrophy, progressive supranuclear palsy, corticobasal degeneration, and dementia with Lewy bodies, is degeneration of the presynaptic dopaminergic pathways in the basal ganglia. Conventional MR imaging is insensitive, especially in early disease, so functional imaging has become the primary method used to differentiate a true parkinsonian syndrome from vascular parkinsonism, drug-induced changes, or essential tremor. Unusually for a modern functional imaging technique, the method most widely used in European clinics depends on SPECT and not PET. This SPECT technique (described in the first of 2 parts) commonly reports dopamine-transporter function, with decreasing striatal uptake demonstrating increasingly severe disease.

The role of functional dopamine-transporter SPECT imaging in parkinsonian syndromes, part 2.

SUMMARY: The functional imaging technique most widely used in European clinics to differentiate a true parkinsonian syndrome from vascular parkinsonism, drug-induced changes, or essential tremor is dopamine-transporter SPECT. This technique commonly reports dopamine-transporter function, with decreasing striatal uptake demonstrating increasingly severe disease. The strength of dopamine-transporter SPECT is that nigrostriatal degeneration is observed in both clinically inconclusive parkinsonism and early, even premotor, disease. In this clinical review (Part 2), we present the dopamine-transporter SPECT findings in a variety of neurodegenerative diseases, including multiple system atrophy, progressive supranuclear palsy, corticobasal degeneration, and dementia with Lewy bodies. The findings in vascular parkinsonism, drug-induced parkinsonism, and essential tremor are also described. It is hoped that this technique will be the forerunner of a range of routinely used, process-specific ligands that can identify early degenerative disease and subsequently guide disease-modifying interventions.

In vivo and in vitro correction of the mdx dystrophin gene nonsense mutation by short-fragment homologous replacement.

Targeted genetic correction of mutations in cells is a potential strategy for treating human conditions that involve nonsense, missense, and transcriptional splice junction mutations. One method of targeted gene repair, single-stranded short-fragment homologous replacement (ssSFHR), has been successful in repairing the common deltaF508 3-bp microdeletion at the cystic fibrosis transmembrane conductance regulator (CFTR) locus in 1% of airway epithelial cells in culture. This study investigates in vitro and in vivo application of a double-stranded method variant of SFHR gene repair to the mdx mouse model of Duchenne muscular dystrophy (DMD). A 603-bp wild-type PCR product was used to repair the exon 23 C-to-T mdx nonsense transition at the Xp21.1 dys locus in cultured myoblasts and in tibialis anterior (TA) from male mdx mice. Multiple transfection and variation of lipofection reagent both improved in vitro SFHR efficiency, with successful conversion of mdx to wild-type nucleotide at the dys locus achieved in 15 to 20% of cultured loci and in 0.0005 to 0.1% of TA. The genetic correction of mdx myoblasts was shown to persist for up to 28 days in culture and for at least 3 weeks in TA. While a high frequency of in vitro gene repair was observed, the lipofection used here appeared to have adverse effects on subsequent cell viability and corrected cells did not express dystrophin transcript. With further improvements to in vitro and in vivo gene repair efficiencies, SFHR may find some application in DMD and other genetic neuromuscular disorders in humans.

CD45 fraction bone marrow cells as potential delivery vehicles for genetically corrected dystrophin loci.

Targeted correction of mutations in muscle can be delivered by direct i.m. injection of corrective DNA to the dystrophic muscle or by autologous injection of cells that have been genetically corrected after isolation from the individual with the dystrophic muscle. The successful application of chimeraplasty and short fragment homologous replacement to correct the exon 23 nonsense mdx transition at the mouse dys locus has opened up the possibility that with further development, targeted gene correction may have some future application for the treatment of muscular dystrophies. In vitro, application of targeted gene correction at the mdx dys locus results in better correction efficiencies than when applied directly to dystrophic muscle. This suggests that at least for the time being, a strategy involving ex vivo correction may be advantageous over a direct approach for delivery of gene correction to dystrophic muscle. This, particularly in view of recent developments indicating that bone-marrow-derived cells are able to systemically remodel dystrophic muscle, whilst penetration of DNA introduced to muscle is limited to individually injected muscles. Application of targeted gene correction to Duchenne dystrophy needs to account for the fact that about 65% of Duchenne muscular dystrophy cases involve large frame-shift deletion of gene sequence at the dys locus. Traditionally, whilst targeted gene correction is able to restore point mutations entirely, it remains to be seen as to whether a strategy for the 'correction' of frame shift deletions may be engineered successfully. This communication discusses the possibility of applying targeted gene correction to dystrophic muscle in Duchenne dystrophy.

Lack of C-erbB-2 protein expression in pulmonary carcinoid tumours.

To determine if amplification of the C-erb-B2 proto-oncogene could be correlated with prognosis in carcinoid tumours, 49 pulmonary carcinoid tumours (26 typical, 23 atypical) were examined using a polyclonal antibody to the C-terminal peptide of the C-erb-B2 protein sequence. No C-erb-B2 gene product could be shown: the demonstration of C-erb-B2 does not seem to help, therefore, in determining diagnosis or prognosis in pulmonary carcinoid tumours.

Detection of MELAS A3243G point mutation in muscle, blood and hair follicles.

Polymerase chain reaction (PCR) based methods for the diagnosis and screening of the mitochondrial disorders have been well established. A number of tissues are routinely used. In this study, we compared the detection rate for MELAS A3243G point mutation in muscle, blood and hair follicles. Ten subjects were studied; mean age was 47 years, (SD 16, range 23-73). All ten subjects had the MELAS A3243G point mutation detected in muscle and hair follicles, but only five had the abnormality in blood samples. The rate of detection of the point mutation in blood samples was age dependent. MtDNA analysis on hair follicles is as sensitive as muscle in detecting this mutation. Analysis using blood samples is not as sensitive, particularly in older subjects. The absence of the mutation in blood samples suggests that there is a preferential selection process for normal (wild type) mtDNA over time. This may be related to the rate of cell division and energy requirements of each tissue.

The polymerase chain reaction in the study of mitochondrial genetics.

Since its development in the late 1980's, the polymerase chain reaction (PCR) has revolutionised molecular genetic studies. It has provided direct access to genetic material in quantities sufficient for meaningful analyses to be performed. Adaptations to the basic technique have resulted in a wide range of applications from basic gene amplification to the estimation of DNA species quantities within cells. The study of human mitochondrial genetics is but one of the many disciplines to benefit from the rapid ascension of PCR based technology. In this communication we outline several uses of the PCR technique in the detection, quantification and characterisation of human mitochondrial genetic defects. The data presented in this communication highlight the versatility and applicability of PCR not only to mitochondrial research but to other disciplines of medical research.

Changes in drug use and HIV/AIDS risk-taking 1989-90.

The Australian National AIDS and Injecting Drug Use Study was designed to monitor the risk behaviour of Australian injecting drug users (IDUs) in a number of major cities, and to estimate the seroprevalence of those interviewed. Differences in risk behaviour found across 2 years in Perth are reported. One hundred and ninety-six Perth IDUs were interviewed in 1989, and 150 in 1990 using the same survey questionnaire, with a small cohort of 38 respondents being followed up across the 2 years.Significant injecting differences between 1989 and 1990 in both the cohort and independent samples were found. In general, these amounted to a greater likelihood that a new needle and syringe would be used on each injecting occasion, and a greater use of bleach in 1990 than in 1989. The most common response of respondents who said they had changed their drug use behaviour in 1990 was to report ceasing to share needles, while the most common response in 1989 was reduced sharing. There was an increased use of condoms for vaginal intercourse with all partners in both studies across the 2 years. Seropositivity for all new cases across the 2 years was 1.75%. It is apparent that there has been a significant shift in risky behaviour in the direction of greater safety across the 2 years. There is also objective corroborative evidence in increased demand for sterile injecting equipment sold through pharmacists, although there is no evidence that drug use increased significantly during the same period. It is concluded that the availability of sterile needles and syringes should be maintained and improved and that emphasis should be placed on encouraging safer sexual behaviour among IDUs.

An evaluation of a primary care psychological therapies clinic.

The present study evaluates a primary care psychological therapies clinic providing input one day per week to a GP practice in a rural area of Scotland. Routine data was collected over a 17 month period and a rating scale was used to evaluate treatment outcomes by both GPs and therapist. In addition GP satisfaction with the service was also evaluated. No significant differences were found between counselling and psychology cases on any of the variables examined. Treatment outcome evaluation showed an overall rate of improvement following treatment, with a mean score of 7 on a 1-10 scale, with 10 representing 'much better'. Significant agreement was found between therapist and GP on rating outcome and no significant differences were found between ratings given by psychologist and counsellor. The GPs involved also expressed high levels of satisfaction with the service.

The accurate measurement of second virial coefficients using self-interaction chromatography: experimental considerations.

Measurement of B22, the second virial coefficient, is an important technique for describing the solution behaviour of proteins, especially as it relates to precipitation, aggregation and crystallisation phenomena. This paper describes the best practise for calculating B22 values from self-interaction chromatograms (SIC) for aqueous protein solutions. Detailed analysis of SIC peak shapes for lysozyme shows that non-Gaussian peaks are commonly encountered for SIC, with typical peak asymmetries of 10%. This asymmetry reflects a non-linear chromatographic retention process, in this case heterogeneity of the protein-protein interactions. Therefore, it is important to use the centre of mass calculations for determining accurate retention volumes and thus B22 values. Empirical peak maximum chromatogram analysis, often reported in the literature, can result in errors of up to 50% in B22 values. A methodology is reported here for determining both the mean and the variance in B22 from SIC experiments, includes a correction for normal longitudinal peak broadening. The variance in B22 due to chemical effects is quantified statistically and is a measure of the heterogeneity of protein-protein interactions in solution. In the case of lysozyme, a wide range of B22 values are measured which can vary significantly from the average B22 values.

Engineering a multimodal nerve conduit for repair of injured peripheral nerve.

Injury to nerve tissue in the peripheral nervous system (PNS) results in long-term impairment of limb function, dysaesthesia and pain, often with associated psychological effects. Whilst minor injuries can be left to regenerate without intervention and short gaps up to 2 cm can be sutured, larger or more severe injuries commonly require autogenous nerve grafts harvested from elsewhere in the body (usually sensory nerves). Functional recovery is often suboptimal and associated with loss of sensation from the tissue innervated by the harvested nerve. The challenges that persist with nerve repair have resulted in development of nerve guides or conduits from non-neural biological tissues and various polymers to improve the prognosis for the repair of damaged nerves in the PNS. This study describes the design and fabrication of a multimodal controlled pore size nerve regeneration conduit using polylactic acid (PLA) and (PLA):poly(lactic-co-glycolic) acid (PLGA) fibers within a neurotrophin-enriched alginate hydrogel. The nerve repair conduit design consists of two types of PLGA fibers selected specifically for promotion of axonal outgrowth and Schwann cell growth (75:25 for axons; 85:15 for Schwann cells). These aligned fibers are contained within the lumen of a knitted PLA sheath coated with electrospun PLA nanofibers to control pore size. The PLGA guidance fibers within the nerve repair conduit lumen are supported within an alginate hydrogel impregnated with neurotrophic factors (NT-3 or BDNF with LIF, SMDF and MGF-1) to provide neuroprotection, stimulation of axonal growth and Schwann cell migration. The conduit was used to promote repair of transected sciatic nerve in rats over a period of 4 weeks. Over this period, it was observed that over-grooming and self-mutilation (autotomy) of the limb implanted with the conduit was significantly reduced in rats implanted with the full-configuration conduit compared to rats implanted with conduits containing only an alginate hydrogel. This indicates return of some feeling to the limb via the fully-configured conduit. Immunohistochemical analysis of the implanted conduits removed from the rats after the four-week implantation period confirmed the presence of myelinated axons within the conduit and distal to the site of implantation, further supporting that the conduit promoted nerve repair over this period of time. This study describes the design considerations and fabrication of a novel multicomponent, multimodal bio-engineered synthetic conduit for peripheral nerve repair.

Sentinel node studies in truncal melanoma: does an increased number of draining basins correlate with an increased risk of lymph metastasis?

OBJECTIVES: To assess whether an association exists between drainage to multiple basins and lymphatic metastasis in patients with truncal melanoma. METHODS: We retrospectively reviewed 227 patients with primary malignant melanoma between January 2006 and December 2009. All patients received an intradermal injection of (99m)Tc-nanocolloid and lymphoscintigraphy followed by sentinel node biopsy. Pre-staging histology with Breslow thickness from excision biopsy was also obtained. RESULTS: 82/227 (36%) patients with primary truncal melanoma were identified. Nodal histology was positive for metastatic disease in 27/82 (32.9%) patients. Of these 27, 15 had 1 basin of drainage, 7 had 2 basins of drainage and 5 had 3 basins of drainage. Of the 55 node-negative patients, 35 had 1 basin, 18 had 2 basins and 2 had 3 basins of drainage. We found no significant correlation with sentinel node positivity and those that had ≥2 drainage basins. Breslow thickness was available in 65/82(79.2%) patients. Sentinel node biopsy was positive in 6/28 patients who had <1.5 mm thickness, 8/14 who had a 1.5-3.9 mm thickness and 9/23 who had ≥4 mm thickness. There was a significant correlation between Breslow thickness of ≥4 mm and nodal positivity (P = 0.03). CONCLUSION: This study demonstrates no association between multiple drainage basins and sentinel node histology. Sentinel lymph node status did correlate with Breslow thickness.