HIV broadly neutralizing antibody escapability drives the therapeutic efficacy of vectored immunotherapy.

Broadly neutralizing antibodies (bNAbs) have shown great promise for prevention and treatment of HIV infection. Breadth of bNAb neutralization, measured in vitro across panels of diverse viral isolates, is often used as a predictor of clinical potential. However, recent prevention studies demonstrate that the clinical efficacy of a broad and potent bNAb (VRC01) is undermined by neutralization resistance of circulating strains. Using HIV-infected humanized mice, we find that therapeutic efficacy of bNAbs delivered as Vectored ImmunoTherapy (VIT) is a function of both the fitness cost and resistance benefit of mutations that emerge during viral escape, which we term 'escapability'. Applying this mechanistic framework, we find that the sequence of the envelope V5-loop alters the resistance benefits of mutants that arise during escape, thereby impacting the therapeutic efficacy of VIT-mediated viral suppression. We also find that an emtricitabine-based antiretroviral drug regimen dramatically enhances the efficacy of VIT, by reducing the fitness of mutants along the escape path. Our findings demonstrate that bNAb escapability is a key determinant to consider in the rational design of antibody regimens with maximal efficacy and illustrates a tractable means of minimizing viral escape from existing bNAbs.

Digital wearable insole-based identification of knee arthropathies and gait signatures using machine learning.

Gait is impaired in musculoskeletal conditions, such as knee arthropathy. Gait analysis is used in clinical practice to inform diagnosis and monitor disease progression or intervention response. However, clinical gait analysis relies on subjective visual observation of walking as objective gait analysis has not been possible within clinical settings due to the expensive equipment, large-scale facilities, and highly trained staff required. Relatively low-cost wearable digital insoles may offer a solution to these challenges. In this work, we demonstrate how a digital insole measuring osteoarthritis-specific gait signatures yields similar results to the clinical gait-lab standard. To achieve this, we constructed a machine learning model, trained on force plate data collected in participants with knee arthropathy and controls. This model was highly predictive of force plate data from a validation set (area under the receiver operating characteristics curve [auROC] = 0.86; area under the precision-recall curve [auPR] = 0.90) and of a separate, independent digital insole dataset containing control and knee osteoarthritis subjects (auROC = 0.83; auPR = 0.86). After showing that digital insole-derived gait characteristics are comparable to traditional gait measurements, we next showed that a single stride of raw sensor time-series data could be accurately assigned to each subject, highlighting that individuals using digital insoles can be identified by their gait characteristics. This work provides a framework for a promising alternative to traditional clinical gait analysis methods, adds to the growing body of knowledge regarding wearable technology analytical pipelines, and supports clinical development of at-home gait assessments, with the potential to improve the ease, frequency, and depth of patient monitoring.

The way we walk ­ our 'gait' ­ is a key indicator of health. Gait irregularities like limping, shuffling or a slow pace can be signs of muscle or joint problems. Assessing a patient's gait is therefore an important element in diagnosing these conditions, and in evaluating whether treatments are working. Gait is often assessed via a simple visual inspection, with patients being asked to walk back and forth in a doctor's office. While quick and easy, this approach is highly subjective and therefore imprecise. 'Objective gait analysis' is a more accurate alternative, but it relies on tests being conducted in specialised laboratories with large-scale, expensive equipment operated by highly trained staff. Unfortunately, this means that gait laboratories are not accessible for everyday clinical use. In response, Wipperman et al. aimed to develop a low-cost alternative to the complex equipment used in gait laboratories. To do this, they harnessed wearable sensor technologies ­ devices that can directly measure physiological data while embedded in clothing or attached to the user. Wearable sensors have the advantage of being cheap, easy to use, and able to provide clinically useful information without specially trained staff. Wipperman et al. analysed data from classic gait laboratory devices, as well as 'digital insoles' equipped with sensors that captured foot movements and pressure as participants walked. The analysis first 'trained' on data from gait laboratories (called force plates) and then applied the method to gait measurements obtained from digital insoles worn by either healthy participants or patients with knee problems. Analysis of the pressure data from the insoles confirmed that they could accurately predict which measurements were from healthy individuals, and which were from patients. The gait characteristics detected by the insoles were also comparable to lab-based measurements ­ in other words, the insoles provided similar type and quality of data as a gait laboratory. Further analysis revealed that information from just a single step could reveal additional information about the subject's walking. These results support the use of wearable devices as a simple and relatively inexpensive way to measure gait in everyday clinical practice, without the need for specialised laboratories and visits to the doctor's office. Although the digital insoles will require further analytical and clinical study before they can be widely used, Wipperman et al. hope they will eventually make monitoring muscle and joint conditions easier and more affordable.

Chromosome-level genome assembly of the aster leafhopper (Macrosteles quadrilineatus) reveals the role of environment and microbial symbiosis in shaping pest insect genome evolution.

Leafhoppers comprise over 20,000 plant-sap feeding species, many of which are important agricultural pests. Most species rely on two ancestral bacterial symbionts, Sulcia and Nasuia, for essential nutrition lacking in their phloem and xylem plant sap diets. To understand how pest leafhopper genomes evolve and are shaped by microbial symbioses, we completed a chromosomal-level assembly of the aster leafhopper's genome (ALF; Macrosteles quadrilineatus). We compared ALF's genome to three other pest leafhoppers, Nephotettix cincticeps, Homalodisca vitripennis, and Empoasca onukii, which have distinct ecologies and symbiotic relationships. Despite diverging ~155 million years ago, leafhoppers have high levels of chromosomal synteny and gene family conservation. Conserved genes include those involved in plant chemical detoxification, resistance to various insecticides, and defence against environmental stress. Positive selection acting upon these genes further points to ongoing adaptive evolution in response to agricultural environments. In relation to leafhoppers' general dependence on symbionts, species that retain the ancestral symbiont, Sulcia, displayed gene enrichment of metabolic processes in their genomes. Leafhoppers with both Sulcia and its ancient partner, Nasuia, showed genomic enrichment in genes related to microbial population regulation and immune responses. Finally, horizontally transferred genes (HTGs) associated with symbiont support of Sulcia and Nasuia are only observed in leafhoppers that maintain symbionts. In contrast, HTGs involved in non-symbiotic functions are conserved across all species. The high-quality ALF genome provides deep insights into how host ecology and symbioses shape genome evolution and a wealth of genetic resources for pest control targets.

Phylloxera and Aphids Show Distinct Features of Genome Evolution Despite Similar Reproductive Modes.

Genomes of aphids (family Aphididae) show several unusual evolutionary patterns. In particular, within the XO sex determination system of aphids, the X chromosome exhibits a lower rate of interchromosomal rearrangements, fewer highly expressed genes, and faster evolution at nonsynonymous sites compared with the autosomes. In contrast, other hemipteran lineages have similar rates of interchromosomal rearrangement for autosomes and X chromosomes. One possible explanation for these differences is the aphid's life cycle of cyclical parthenogenesis, where multiple asexual generations alternate with 1 sexual generation. If true, we should see similar features in the genomes of Phylloxeridae, an outgroup of aphids which also undergoes cyclical parthenogenesis. To investigate this, we generated a chromosome-level assembly for the grape phylloxera, an agriculturally important species of Phylloxeridae, and identified its single X chromosome. We then performed synteny analysis using the phylloxerid genome and 30 high-quality genomes of aphids and other hemipteran species. Unexpectedly, we found that the phylloxera does not share aphids' patterns of chromosome evolution. By estimating interchromosomal rearrangement rates on an absolute time scale, we found that rates are elevated for aphid autosomes compared with their X chromosomes, but this pattern does not extend to the phylloxera branch. Potentially, the conservation of X chromosome gene content is due to selection on XO males that appear in the sexual generation. We also examined gene duplication patterns across Hemiptera and uncovered horizontal gene transfer events contributing to phylloxera evolution.

Phylloxera and aphids show distinct features of genome evolution despite similar reproductive modes.

Genomes of aphids (family Aphididae) show several unusual evolutionary patterns. In particular, within the XO sex determination system of aphids, the X chromosome exhibits a lower rate of interchromosomal rearrangements, fewer highly expressed genes, and faster evolution at nonsynonymous sites compared to the autosomes. In contrast, other hemipteran lineages have similar rates of interchromosomal rearrangement for autosomes and X chromosomes. One possible explanation for these differences is the aphid's life cycle of cyclical parthenogenesis, where multiple asexual generations alternate with one sexual generation. If true, we should see similar features in the genomes of Phylloxeridae, an outgroup of aphids which also undergoes cyclical parthenogenesis. To investigate this, we generated a chromosome-level assembly for the grape phylloxera, an agriculturally important species of Phylloxeridae, and identified its single X chromosome. We then performed synteny analysis using the phylloxerid genome and 30 high-quality genomes of aphids and other hemipteran species. Unexpectedly, we found that the phylloxera does not share aphids' patterns of chromosome evolution. By estimating interchromosomal rearrangement rates on an absolute time scale, we found that rates are elevated for aphid autosomes compared to their X chromosomes, but this pattern does not extend to the phylloxera branch. Potentially, the conservation of X chromosome gene content is due to selection on XO males that appear in the sexual generation. We also examined gene duplication patterns across Hemiptera and uncovered horizontal gene transfer events contributing to phylloxera evolution.

Behavioural determinants of COVID-19 vaccination acceptance among healthcare workers: a rapid review.

OBJECTIVES: Vaccinating healthcare workers (HCWs) against COVID-19 has been a public health priority since rollout began in late 2020. Promoting COVID-19 vaccination among HCWs would benefit from identifying modifiable behavioural determinants. We sought to identify and categorize studies looking at COVID-19 vaccination acceptance to identify modifiable factors to increase uptake in HCWs. STUDY DESIGN: Rapid evidence review. METHODS: We searched MEDLINE and Cochrane databases until May 2021 and conducted a grey literature search to identify cross-sectional, cohort, and qualitative studies. Key barriers to, and enablers of, vaccine acceptance were categorized using the Theoretical Domains Framework (TDF), a comprehensive theoretical framework comprising 14 behavioural domains. RESULTS: From 19,591 records, 74 studies were included. Almost two-thirds of responding HCWs were willing to accept a COVID-19 vaccine (median = 64%, interquartile range = 50-78%). Twenty key barriers and enablers were identified and categorized into eight TDF domains. The most frequently identified barriers to COVID-19 vaccination were as follows: concerns about vaccine safety, efficacy, and speed of development (TDF domain: Beliefs about consequences); individuals in certain HCW roles (Social/professional role and identity); and mistrust in state/public health response to COVID-19 (Social influences). Routinely being vaccinated for seasonal influenza (Reinforcement), concerns about contracting COVID-19 (Beliefs about consequences) and working directly with COVID-19 patients (Social/professional role and identity) were key enablers of COVID-19 vaccination among HCWs. DISCUSSION: Our review identified eight (of a possible 14) behavioural determinants of COVID-19 vaccine acceptance among HCWs that, if targeted, could help design tailored vaccination messaging, policy, campaigns, and programs to support HCWs vaccination uptake.

The Genomic Basis of Evolutionary Novelties in a Leafhopper.

Evolutionary innovations generate phenotypic and species diversity. Elucidating the genomic processes underlying such innovations is central to understanding biodiversity. In this study, we addressed the genomic basis of evolutionary novelties in the glassy-winged sharpshooter (Homalodisca vitripennis, GWSS), an agricultural pest. Prominent evolutionary innovations in leafhoppers include brochosomes, proteinaceous structures that are excreted and used to coat the body, and obligate symbiotic associations with two bacterial types that reside within cytoplasm of distinctive cell types. Using PacBio long-read sequencing and Dovetail Omni-C technology, we generated a chromosome-level genome assembly for the GWSS and then validated the assembly using flow cytometry and karyotyping. Additional transcriptomic and proteomic data were used to identify novel genes that underlie brochosome production. We found that brochosome-associated genes include novel gene families that have diversified through tandem duplications. We also identified the locations of genes involved in interactions with bacterial symbionts. Ancestors of the GWSS acquired bacterial genes through horizontal gene transfer (HGT), and these genes appear to contribute to symbiont support. Using a phylogenomics approach, we inferred HGT sources and timing. We found that some HGT events date to the common ancestor of the hemipteran suborder Auchenorrhyncha, representing some of the oldest known examples of HGT in animals. Overall, we show that evolutionary novelties in leafhoppers are generated by the combination of acquiring novel genes, produced both de novo and through tandem duplication, acquiring new symbiotic associations that enable use of novel diets and niches, and recruiting foreign genes to support symbionts and enhance herbivory.

Glutaminase 2 knockdown reduces hyperammonemia and associated lethality of urea cycle disorder mouse model.

Amino acids, the building blocks of proteins in the cells and tissues, are of fundamental importance for cell survival, maintenance, and proliferation. The liver plays a critical role in amino acid metabolism and detoxication of byproducts such as ammonia. Urea cycle disorders with hyperammonemia remain difficult to treat and eventually necessitate liver transplantation. In this study, ornithine transcarbamylase deficient (Otcspf-ash ) mouse model was used to test whether knockdown of a key glutamine metabolism enzyme glutaminase 2 (GLS2, gene name: Gls2) or glutamate dehydrogenase 1 (GLUD1, gene name: Glud1) could rescue the hyperammonemia and associated lethality induced by a high protein diet. We found that reduced hepatic expression of Gls2 but not Glud1 by AAV8-mediated delivery of a short hairpin RNA in Otcspf-ash mice diminished hyperammonemia and reduced lethality. Knockdown of Gls2 but not Glud1 in Otcspf-ash mice exhibited reduced body weight loss and increased plasma glutamine concentration. These data suggest that Gls2 hepatic knockdown could potentially help alleviate risk for hyperammonemia and other clinical manifestations of patients suffering from defects in the urea cycle.

Identification of sex chromosomes and primary sex ratio in the small hive beetle, a worldwide parasite of honey bees.

BACKGROUND: The small hive beetle (SHB), Aethina tumida, has emerged as a worldwide threat to honey bees in the past two decades. These beetles harvest nest resources, feed on larval bees, and ultimately spoil nest resources with gelatinous slime together with the fungal symbiont Kodamaea ohmeri. RESULTS: Here, we present the first chromosome-level genome assembly for the SHB. With a 99.1% representation of conserved (BUSCO) arthropod genes, this resource enables the study of chemosensory, digestive, and detoxification traits critical for SHB success and possible control. We use this annotated assembly to characterize features of SHB sex chromosomes and a female-skewed primary sex ratio. We also found chromosome fusion and a lower recombination rate in sex chromosomes than in autosomes. CONCLUSIONS: Genome-enabled insights will clarify the traits that allowed this beetle to exploit hive resources successfully and will be critical for determining the causes of observed sex ratio asymmetries.

Investing to prevent: Description of an innovative approach to commissioning a supervised toothbrushing programme across multiple local authorities in England.

INTRODUCTION: The NHS Long Term Plan prioritises NHS action to reduce health inequalities and give children a good start in life. A Sustainability and Transformation Partnership (STP) is a collaborative working arrangement between local authorities and the NHS covering a defined population and geography. Within the STP in Devon, England, all three local authorities had separate supervised toothbrushing programmes; all were precariously funded. Devon has limited access to routine NHS dentistry and children in deprived areas have high rates of general anaesthetics for dental extractions. Consolidating the supervised toothbrushing programmes presented an opportunity to address oral health inequalities across Devon STP. OBJECTIVES: 1. Reduce oral health inequalities for children in deprived areas. 2. Reduce treatment need for children who have limited access to routine NHS dentistry. 3. Invest in prevention. METHODS: A proposal, supported by local authorities in Devon STP, was developed for a targeted supervised toothbrushing programme at early years sites across the most deprived 50% of areas in Devon. Return on investment was estimated using a national resource. Methods are described for identifying eligible sites and defining procurement lots. The NHS dental services commissioner agreed to support this proposal using an innovative approach to commissioning. RESULTS: Three lots, totalling 525 sites, were awarded to two providers. Mobilisation over summer 2019 led to implementation from September 2019. CONCLUSION: Partnership working and innovative commissioning can enable NHS England to invest in prevention at scale where options to increase dental access are limited. Implementation across a large geographical area creates challenges but facilitates equitable programme delivery.

Dentists' perceptions of their professional roles regarding referrals within primary dental care in England: a qualitative study.

OBJECTIVE: To describe dentists' perceptions of their professional roles, including the reasons why they make, accept or decline patient referrals within primary dental care in England. BASIC RESEARCH DESIGN: Qualitative semi-structured interviews, conducted via Skype, telephone or face-to-face. Transcripts were analysed using thematic analysis and typologies were developed. PARTICIPANTS: Ten general dental practitioners (GDPs) and 12 community dentists working in England. RESULTS: Five main themes were identified: professional independence, the nature of dental care, the business of dentistry, obscure rules and 'no man's land'. This final theme described a notional gap between GDPs' and community dentists' responsibilities towards vulnerable people, who were perceived by participants to include frail older people, anxious and socially marginalised adults and children with high levels of disease. Three typologies of dentists were generated. 'Entrepreneurs' felt no allegiance to the National Health Service and no obligation to treat vulnerable patients. 'Altruistic carers' were committed to caring for exceptionally deserving patients. 'Pragmatic carers' tried to provide relational dental care (time and emotional support) for vulnerable patients but encountered discouraging systemic barriers. CONCLUSION: Dentists' perceptions of their roles may influence whether and how they provide access to primary dental care for vulnerable people through referral systems. Access issues may exacerbate the oral health inequalities experienced by vulnerable groups. Based upon the findings, approaches are proposed that may encourage and enable the dental workforce to support vulnerable people actively to receive primary dental care.

Expression of SREBP2 and cholesterol metabolism related genes in TCGA glioma cohorts.

Diffuse gliomas are the most common primary brain tumors. The Cancer Genome Atlas (TCGA) database provides correlative evidence between altered molecular pathways and gliomas. Dysregulated cholesterol homeostasis emerges as a potential indicator of the pathogenesis of gliomas.Mining large cohorts from the TCGA together with database from the Chinese Glioma Genome Atlas (CGGA) for confirmation, we compared gene expression of cholesterol synthesis master regulator SREBP2 and its regulatory networks in low grade glioma (LGG) and glioblastoma (GBM).Our analysis shows that expression of SREBP2 and related genes is lower in GBM than in LGG, indicating that cholesterol metabolism processes, including de novo synthesis, cholesterol uptakes, and cholesterol conversion and efflux, are suppressed in GBM.Overall, our data suggests that SREBP2 transcript could serve as a potential prognosis marker or therapeutic target in diffuse glioma including GBM.

LuceDex: a prospective study comparing ranibizumab plus dexamethasone combination therapy versus ranibizumab monotherapy for neovascular age-related macular degeneration.

BACKGROUND: The LuceDex prospective randomized pilot trial compared the combination of intravitreal ranibizumab and dexamethasone with ranibizumab monotherapy for treatment of neovascular age-related macular degeneration. METHODS: Thirty-seven eyes of 37 patients were randomized 1:1 between combination therapy with intravitreal ranibizumab and dexamethasone (Group 1) and intravitreal ranibizumab monotherapy (Group 2). All study eyes received 4 monthly treatments followed by monthly treatment on indication. RESULTS: In the LuceDex study, eyes gained an average of 11.1 and 5.9 Early Treatment of Diabetic Retinopathy Study letters in Groups 1 and 2, respectively, at Month 12. No more than zero Early Treatment of Diabetic Retinopathy Study letters were lost in 88% of Group 1 eyes and 70% of Group 2 eyes. The average number of treatments per study eye by Month 12 was 7.1 in Group 1 and 6.6 in Group 2. Choroidal neovascular membrane size decreased in Group 1 significantly compared with Group 2 (P < 0.05). CONCLUSION: The LuceDex pilot study suggested a possible benefit of adding intravitreal dexamethasone to treatment of neovascular age-related macular degeneration with intravitreal ranibizumab. A larger study is needed to further identify and define possible benefits of this combination therapy.

Neonatal infraorbital nerve transection in rat results in peripheral trigeminal sprouting.

Retrograde tracing techniques were employed to determine whether transection of the infraorbital (IO) nerve in either newborn or adult rats resulted in peripheral sprouting by undamaged trigeminal (V) axons. The IO nerve was sectioned just behind the vibrissa pad, either on the day of birth or when animals reached at least 60 days of age. After an additional 60 days, the same nerve was retransected in the orbit; horseradish peroxidase (HRP) or diamidino yellow (DY) was injected into the central portion of the vibrissa pad; and animals were killed 2-3 days later. In the neonatally nerve-damaged rats, this procedure invariably labelled primary afferent neurons in both the ipsilateral and contralateral V ganglia. On the ipsilateral side, these cells were located in the caudal portion of the ophthalmic-maxillary region and, less often, in the mandibular division. Their average diameter was 22.6 micron (s.d. = 5.6). On the contralateral side, most labelled ganglion cells were visible in the anteromedial part of the ophthalmic-maxillary region but a few could also be seen in the mandibular division. Their average diameter was 21.1 micron (s.d. = 5.5). No labelled ganglion cells were observed in adult rats subjected to the same series of manipulations. In a separate series of neonatally nerve-damaged animals, the above-described procedures were combined with neonatal injection of capsaicin in an effort to determine whether the observed sprouting was dependent upon the presence of large numbers of unmyelinated axons. The addition of this treatment reduced the number of labelled cells in both the ipsilateral and contralateral ganglia, but it did not alter either their distribution or average soma diameter. In a final experiment, sequential double-labelling techniques were used to determine whether the V axons that projected to the vibrissa pad via non-IO nerve branches were the result of sprouting by undamaged ganglion cells or arose from neurons that had originally projected into the IO nerve, were axotomized by our lesions, and regenerated to the vibrissa pad via another V branch. Here, the long-lived retrograde tracer true blue (TB) was injected into the vibrissa pad 6-8 hours before the neonatal nerve cut and DY was deposited into the pad after transection of the regenerate IO nerve in adulthood. Double-labelled cells in this experiment would have projected to the vibrissa pad via the IO nerve at birth and regenerated to it via another V branch in adulthood. Nearly 55% of the DY-labelled cells in this experiment also contained TB.(ABSTRACT TRUNCATED AT 400 WORDS)

The projection from the superficial to the deep layers of the superior colliculus: an intracellular horseradish peroxidase injection study in the hamster.

Intracellular recording and horseradish peroxidase (HRP) injection techniques were employed to examine the projections of superficial layer [stratum griseum superficiale (SGS) and stratum opticum (SO)] superior collicular (SC) neurons in the hamster that sent axon collaterals into the deep laminae (those ventral to the SO) of this structure. Sixty-nine neurons were studied, selected from a sample of over 185 HRP-filled superficial layer cells on the basis of having heavily stained axons. Of the 69 cells included in the study, 43.4% (n = 30) sent at least one axon collateral to the deep laminae. Not all cell types in the superficial layers contributed equally to this interlaminar projection: 78.6% (n = 11) of the recovered wide-field vertical cells, 55.0% (n = 11) of the narrow-field vertical cells, 16.7% (n = 2) of the stellate cells, 40.0% (n = 2) of the marginal cells, 18.2% (n = 2) of the horizontal cells, and 28.6% (n = 2) of neurons we could not classify on the basis of their somadendritic morphology projected to the deep layers. Within a given cell class, there were no significant morphological or physiological differences between the neurons that possessed deep axon collaterals and those that did not. The deep axon collaterals of most of the interlaminar projection neurons were restricted to the stratum griseum intermediate (SGI). In this layer, the largest segment of the axon arbor was located lateral to a projection line that was orthogonal to the SC surface and that passed through the soma of the cell in question. These results, along with those of a previous study (Mooney et al., 1984), which demonstrated that the dendrites of deep layer cells may extend through the SO and into the SGS, indicate that there is an extensive anatomical substrate by which sensory information may be communicated from superficial to deep layer SC neurons.

Prostaglandin synthesis inhibitors block alcohol-induced fetal hypoplasia.

Alcohol-induced growth retardation is a fetal effect consistently associated with maternal ethanol consumption. In humans, those infants whose mothers consume even a limited amount of ethanol during pregnancy have a significant incidence of growth inhibition. The molecular mechanism responsible for this growth deficiency is unknown, and prevention depends on maternal abstinence during pregnancy. The data reported here suggest that ethanol-mediated increases in tissue prostaglandin (PG) E levels (PGE1 plus PGE2) are correlated with the growth retardation. Further, simultaneous administration of PG synthesis inhibitors with the alcohol blocks the rise in tissue PG levels and protects against the alcohol-induced hypoplasia.

Maintaining the lung disease patient at home.

A pilot home care program for lung disease patients has been set up by a lung association, in conjunction with local hospitals and home care agencies. The emphasis is on long-term care and support of services for which home care agencies would not ordinarily be reimbursed.