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Anterior cruciate ligament (ACL) injuries are a major problem in the pediatric and adolescent populations. Some of these injuries are only partial; yet, there is limited data to inform clinical treatment of such partial tears. It is unknown how injury partial injury impacts long-term degenerative changes in the joint relative to complete injury. In this study, we hypothesized that partial (anteromedial (AM) or posterolateral (PL) bundle) tears would result in small levels of instability and degeneration relative to complete ACL tears and that the degree of degeneration would associate with joint instability. Partial (isolated AM or PL bundle) or complete ACL injury was arthroscopically created in 3-month-old juvenile pigs. The contralateral limb served as a sham-operated control. Six months after injury, joint biomechanics was assessed along with cartilage and meniscus degeneration (via magnetic resonance imaging [MRI], gross imaging, and histology). Joint laxity increases were minimal after PL bundle injury (difference relative to controls (confidence interval): 0.5 (−1.2–2.2) mm), minor after AM bundle injury (3.7 (2.0–5.4) mm), and major after ACL injury (15.8 (13.7–17.8) mm). Cartilage MRI T1ρ relaxation times increased minimally after PL bundle injury (−0.9 (−5.1–3.3) ms for lateral tibia), moderately after AM bundle injury (6.6 (1.7–11.4) ms), and substantially after ACL injury (10.8 (2.1–19.5) ms). Changes in meniscus volume followed a similar rank order. Degeneration was associated with the extent of joint destabilization. These findings suggest that cartilage and meniscus degeneration in the skeletally immature joint are associated with joint laxity after partial and complete ACL injuries. [ABSTRACT FROM AUTHOR]

Azole resistance is a concern for the management of diseases caused by Aspergillus fumigatus in humans. Azole fungicide use in the environment has been identified as a possible cause for development of resistance, which increases the complexity and number of stakeholders involved in this emerging problem. A workshop was held in Amsterdam early 2019 in which stakeholders, including medical and agricultural researchers, representatives from the government, public health, fungicide producers and end-users, reviewed the current evidence supporting environmental selection for resistance and to discuss which research and measures are needed to retain the effectiveness of the azole class for environmental and medical applications. This paper provides an overview of the latest insights and understanding of azole resistance development in the clinical setting and the wider environment. A One Health problem approach was undertaken to list and prioritize which research will be needed to provide missing evidence and to enable preventive interventions. • Environmental azole resistance selection in A. fumigatus threatens medical therapy. • Certain azole fungicide applications have a low risk of resistance selection. • Hotspots involve A. fumigatus growth in plant waste containing azole residues. • Environmental resistance burden may be reduced through a One-Health approach. • AMR research and action plans should include fungal resistance. [ABSTRACT FROM AUTHOR]

Melt electrowriting (MEW) is an additive manufacturing technique that has the potential to create fibrous scaffolds that reproduce the scale and organization of collagen fiber networks in musculoskeletal soft tissues. For musculoskeletal soft tissue engineering, it is useful for scaffolds to have a high aspect ratio (length to width ratio of 5:1 or higher). However, the relationship between MEW process variables and the structural and mechanical properties of such scaffolds is not well understood. In addition, prior studies have cut samples from larger MEW structures, resulting in test specimens with discontinuous fibers. In this study, MEW scaffolds with low (square, 12 mm × 12 mm) and high aspect ratio (rectangular, 35 mm × 5 mm) macroscale geometries were fabricated at varying stage translation speeds or melt extrusion temperatures. Fiber morphology in both geometries and mechanical properties of the continuous rectangular structures were then quantified. Fiber diameter in both square and rectangular scaffolds generally decreased with increasing stage speed, but increased with melt temperature, though the effect of the latter was greater in square scaffolds. Interfiber spacing in both geometries was closer to the intended value as stage speed increased. Spacing became less accurate in square scaffolds with increasing melt temperature but changed little in rectangular scaffolds. Transverse fiber angle in rectangular scaffolds improved with increasing stage speed and had a median value within 1.4% of the intended angle at all temperatures. Finally, apparent tensile modulus in rectangular scaffolds decreased with increasing speed and temperature. These findings highlight the need to tailor MEW process parameters in scaffolds with high aspect ratio geometry in order to consistently generate specific structural and mechanical properties. Because of the potential to reproduce the structural anisotropy, fiber size, and mechanical properties of collagenous extracellular matrix, MEW structures are promising as musculoskeletal soft tissue scaffolds. [ABSTRACT FROM AUTHOR]

The study of infectious disease outbreaks is required to train today’s epidemiologists. A typical way to introduce and explain key epidemiological concepts is through the analysis of a historical outbreak. There are, however, few training options that explicitly utilise real-time simulated stochastic outbreaks where the participants themselves comprise the dataset they subsequently analyse. In this paper, we present a teaching exercise in which an infectious disease outbreak is simulated over a five-day period and subsequently analysed. We iteratively developed the teaching exercise to offer additional insight into analysing an outbreak. An R package for visualisation, analysis and simulation of the outbreak data was developed to accompany the practical to reinforce learning outcomes. Computer simulations of the outbreak revealed deviations from observed dynamics, highlighting how simplifying assumptions conventionally made in mathematical models often differ from reality. Here we provide a pedagogical tool for others to use and adapt in their own settings. [ABSTRACT FROM AUTHOR]

Forensic DNA testing is grounded in molecular biology and population genetics. The technologies that were the basis of restriction length polymorphism testing (RFLP) have given way to PCR based technologies. While PCR has been the pillar of short tandem repeat (STR) methods and will continue to be used as DNA sequencing and analysis of single nucleotide polymorphisms (SNPs) are introduced into human identification, the molecular biology techniques in use today represent significant advances since the introduction of STR testing. Large forensic laboratories with dedicated research teams and forensic laboratories which are part of academic institutions have the resources to keep track of advances which can then be considered for further research or incorporated into current testing methods. However, many laboratories have limited ability to keep up with research advances outside of the immediate area of forensic science and may not have access to a large university library systems. This review focuses on filling this gap with respect to areas of research that intersect with selected methods used in forensic biology. The review summarizes information collected from several areas of the scientific literature where advances in molecular biology have produced information relevant to DNA analysis of sexual assault evidence and methods used in presumptive and confirmatory identification of semen. Older information from the literature is also included where this information may not be commonly known and is relevant to current methods. The topics selected highlight (1) information from applications of proteomics to sperm biology and human reproduction, (2) seminal fluid proteins and prostate cancer diagnostics, (3) developmental biology of sperm from the fertility literature and (4) areas where methods are common to forensic analysis and research in contraceptive use and monitoring. Information and progress made in these areas coincide with the research interests of forensic biology and cross-talk between these disciplines may benefit both. [ABSTRACT FROM AUTHOR]

• SARS-CoV-2 infection can spread rapidly among residents and staff of a long term care facility. • Vaccinations against SARS-CoV-2 is of paramount importance to save lives in US long term care facilities. • Breakthrough SARS-CoV-2 infections in vaccinated long term care facility residents is possible, but a favorable outcome is more likely. An outbreak of SARS-CoV-2 involving four residents of a United States Veterans Affairs long term care facility occurred following administration of the first dose of the mRNA-1273 vaccine (Moderna) to thirty out of 33 residents. Three out of 4 positive cases were partially vaccinated and remained asymptomatic. One of 2 partially vaccinated patients who were tested for anti-spike protein antibodies had detectable levels at the time of diagnosis. The mortality rate was lower compared to a prior outbreak reported in this facility. [ABSTRACT FROM AUTHOR]

Abstract: Pythium insidiosum is an aquatic fungus-like organism in the kingdom Stramenopila that causes pythiosis in both humans and animals. Human pythiosis occurs in ocular, localized granulomatous subcutaneous and systemic or vascular forms. Individuals whose occupations involve exposure to aquatic habitats have an elevated risk of contracting pythiosis. Previously, we reported the first successful isolation of Pythium insidiosum from aquatic environmental samples by culture including confirmation using molecular methods. In this study, we show that P. insidiosum inhabitats moist soil environments in agricultural areas. A total of 303 soil samples were collected from 25 irrigation sources in the areas nearby the recorded home addresses of pythiosis patients residing in northern provinces of Thailand. P. insidiosum DNA was identified directly from each soil extract by using a nested PCR assay and subsequent phylogenetic analysis of the ribosomal intragenic spacer region. P. insidiosum DNA could be detected from 16 of the 25 soil sources (64%). Conventional culture methods were also performed, however all samples exhibited negative culture results. We conclude that both irrigation water and soil are the natural reservoirs of P. insidiosum. In endemic areas, the exposure to these environmental reservoirs should be considered a risk factor for hosts susceptible to pythiosis. [Copyright &y& Elsevier]

Abstract: Penicillium chrysogenum is a ubiquitous airborne fungus detected in every sampled region of the Earth. Owing to its role in Alexander Fleming's serendipitous discovery of Penicillin in 1928, the fungus has generated widespread scientific interest; however its natural history is not well understood. Research has demonstrated speciation within P. chrysogenum, describing the existence of four cryptic species. To discriminate the four species, we developed protocols for species-specific diagnostic PCR directly from fungal conidia. 430 Penicillium isolates were collected to apply our rapid diagnostic tool and explore the distribution of these fungi across the London Underground rail transport system revealing significant differences between Underground lines. Phylogenetic analysis of multiple type isolates confirms that the ‘Fleming species’ should be named Penicillium rubens and that divergence of the four ‘Chrysogenum complex’ fungi occurred about 0.75 million yr ago. Finally, the formal naming of two new species, Penicillium floreyi and Penicillium chainii, is performed. [Copyright &y& Elsevier]

Abstract: Endogenous repair of fibrous connective tissues is limited, and there exist few successful strategies to improve healing after injury. As such, new methods that advance repair by promoting cell growth, extracellular matrix (ECM) production, and tissue integration would represent a marked clinical advance. Using the meniscus as a test platform, we sought to develop an enzyme-releasing scaffold that enhances integrative repair. We hypothesized that the high ECM density and low cellularity of native tissue present physical and biological barriers to endogenous healing, and that localized collagenase treatment might expedite cell migration to the wound edge and tissue remodeling. To test this hypothesis, we fabricated a delivery system in which collagenase was stored inside electrospun poly(ethylene oxide) (PEO) nanofibers and released upon hydration. In vitro results showed that partial digestion of the wound interface improved repair by creating a microenvironment that facilitated cell migration, proliferation and matrix deposition. Specifically, treatment with high-dose collagenase led to a 2-fold increase in cell density at the wound margin and a 2-fold increase in integrative tissue compared to untreated controls at 4weeks (P ⩽0.05). Furthermore, when composite scaffolds containing both collagenase-releasing and structural fiber fractions were placed inside meniscal tears in vitro, enzyme release acted locally and resulted in a positive cellular response similar to that of global treatment with aqueous collagenase. This innovative approach to targeted enzyme delivery may aid the many patients that exhibit meniscal tears by promoting integration of the defect, thereby circumventing the pathologic consequences of partial meniscus removal, and may find widespread application in the treatment of injuries to a variety of dense connective tissues. [Copyright &y& Elsevier]

Abstract: The menisci are crescent-shaped fibrocartilaginous tissues whose structural organization consists of dense collagen bundles that are locally aligned but show a continuous change in macroscopic directionality. This circumferential patterning is necessary for load transmission across the knee joint and is a key design parameter for tissue engineered constructs. To address this issue we developed a novel electrospinning method to produce scaffolds composed of circumferentially aligned (CircAl) nanofibers, quantified their structure and mechanics, and compared them with traditional linearly aligned (LinAl) scaffolds. Fibers were locally oriented in CircAl scaffolds, but their orientation varied considerably as a function of position (P <0.05). LinAl fibers did not change in orientation over a similar length scale (P >0.05). Cell seeding of CircAl scaffolds resulted in a similar cellular directionality. Mechanical analysis of CircAl scaffolds revealed significant interactions between scaffold length and region (P <0.05), with the tensile modulus near the edge of the scaffolds decreasing with increasing scaffold length. No such differences were detected in LinAl specimens (P >0.05). Simulation of the fiber deposition process produced “theoretical” fiber populations that matched the fiber organization and mechanical properties observed experimentally. These novel scaffolds, with spatially varying local orientations and mechanics, will enable the formation of functional anatomic meniscus constructs. [Copyright &y& Elsevier]

Abstract: This article highlights some of the more notable persistent fungal diseases of our times. It draws attention to the emergence of new fungal pathotypes infecting food staple crops, due largely to modern agricultural practices, and to nascent fungal diseases decimating frog populations worldwide and killing hibernating bats in Northern USA. We invoke use of the basic disease triangle concept to highlight the “missing” data, with regards to pathogen and host biology and to the various environmental parameters which may dictate disease spread. Given these data “voids” we comment on the implementation of policy. We conclude with a series of recommendations for improved disease surveillance and reporting, the need for greater public awareness of these issues and a call for greater funding for fungal research. In so doing, we have exploited Magnaporthe oryzae and Batrachochytrium dendrobatidis as exemplar emerging infectious fungi. Our aim is to highlight the impact of emerging and emergent fungi on food security and, more broadly, ecosystem health. [Copyright &y& Elsevier]

Abstract: Extracellular matrix (ECM) bioscaffolds, such as porcine small intestine submucosa (SIS) and urinary bladder matrix (UBM), have been successfully used to improve soft tissue healing. Yet they contain plenty of galactose α1,3 galactose (αGal) epitopes, which cause rejection responses in pig organ transplantation to human. Recently, ECM bioscaffolds derived from genetically modified pigs that are αGal-deficient (αGal(−)) have become available. To ensure that the ECM bioscaffolds from these pigs can be used as alternatives, we examined their morphological, bioactive and biomechanical properties and compared them with those from the wild-type pigs (n =5 per group). Morphologically, the αGal(−) ECMs were found to be similar to the wild-type ECMs in gross observation and matrix appearance with hematoxylin and eosin staining. Growth factors commonly known to be present in ECM bioscaffolds, including FGF-2, TGF-β1, VEGF, IGF-1 and PDGF-BB, also showed no significant differences in terms of quantity (p >0.05) and distribution in tissue from the results of enzyme-linked immunosorbent assay, Western blot analysis and immunohistochemistry. Furthermore, a bromodeoxyuridine cell proliferation assay confirmed the bioactivity of the extracts from the αGal(−) bioscaffolds to be similar to the wild-type bioscaffolds. Under uniaxial tensile testing, no significant differences were found between the αGal(−) and wild-type bioscaffolds in terms of their viscoelastic and mechanical properties (p >0.05). These multidisciplinary results suggest that genetic modification to eliminate the αGal epitopes in the ECM bioscaffolds had not altered the properties of these ECM bioscaffolds and, as such, they should retain their performance in tissue engineering in humans. [Copyright &y& Elsevier]

Abstract: Penicillium marneffei is an endemic mycosis of humans in southeast Asia. Epidemiological data have shown that exposure to soil and season increase the risk of infection, and it is assumed that the main environmental reservoir is in soil. We sampled soils from a P. marneffei - endemic region of Thailand and confirmed by quantitative PCR and sequencing that P. marneffei DNA can be detected, a finding that we replicated over three sampling seasons. P. marneffei-positive and -negative sampling locations can be viewed using a dynamic browser located at www.spatialepidemiology.net/pmarneffei. We subsequently examined the hypothesis that P. marneffei isolates representing the two major phylogeographic clades of this species can grow in: (i) soil and (ii) competition against the closely related species, Talaromyces stipitatus, in a model soil environment. P. marneffei was not detected in non-sterile soil microcosms 14d post inoculation, showing that the pathogen is unable to compete against complete soil fauna under our laboratory conditions. However, both isolates of P. marneffei persisted and increased in biomass when inoculated into sterile soil. P. marneffei stably co-existed with T. stipitatus, and that the main competitive interaction was the inhibition of T. stipitatus growth at low spore application by the ‘Eastern’ isolate of P. marneffei. We conclude that P. marneffei is present in soils within endemic regions, and is able to grow in soil under certain conditions. More research is required to ascertain the specific conditions that regulate the growth of P. marneffei in soils in natural environments. [Copyright &y& Elsevier]

Abstract: Chytridiomycosis is an emerging infectious disease of amphibians caused by the chytrid Batrachochytrium dendrobatidis. The disease has been associated with global amphibian declines and species extinctions, however the principle drivers that underly the emergence of chytridiomycosis remain unclear. Current evidence suggests that the world trade in amphibians is implicated in the emergence of chytridiomycosis. Here, we review the evidence that the amphibian trade is driving the emergence of chytridiomycosis by (1) spreading infected animals worldwide, (2) introducing non-native infected animals into naïve populations and (3) amplifying infection of amphibians by co-housing, followed by untreated discharge of infectious zoospores into water supplies. We conclude that the evidence that the amphibian trade is contributing to the spread of Batrachochytrium dendrobatidis is strong, and that specific actions are necessary to prevent the introduction of the pathogen into thus-far uninfected areas. Specifically, we recommend the development of national risk-abatement plans, focused on firstly preventing introduction of Bd into disease free areas, and secondly, decreasing the impact of the disease on populations that are currently infected. [Copyright &y& Elsevier]

Abstract: The purpose of this in vitro study was to compare the bond strength of various obturation materials to root canal dentin by using a push-out test design. Twenty-five single-rooted extracted human teeth were used in this study. The crowns were removed, and root canal instrumentation was completed by using Endogel-coated 0.06 EndoSequence rotary files (Brasseler, Savannah, GA) and appropriate irrigation with 5.25% NaOCl and a final rinse with 17% EDTA. After instrumentation, the roots were randomly divided into five single-matched cone obturation groups (n = 5 roots/group), as follows: group 1, gutta percha with Kerr EWT (Kerr Corp, Romulus, MI); group 2, gutta percha with AH Plus (Dentsply, DeTrey, Germany); group 3, Resilon (Pentron Clinical Technologies, Wallingford, CT) and Epiphany (Pentron); group 4, Activ GP (Brasseler) obturation system; and group 5, EndoREZ obturation system (Ultradent, South Jordan, UT). The obturated roots were cut perpendicular to the long access to create 1-mm thick slices from the apical, middle, and coronal thirds. The bond strength was measured for each test slice with a push-out testing machine. Statistical analysis was completed by using a two-way analysis of variance and the post hoc Tukeys test with significance set at p < 0.05. Group 2 showed a significantly (p < 0.05) greater bond strength compared with all other groups. Also, groups 1 and 4 had significantly (p < 0.05) higher bond strengths compared with groups 3 and 5. [Copyright &y& Elsevier]