Viral surrogates to screen for virus inactivation (VI) can be a faster, cheaper and safer alternative to third-party testing of pathogenic BSL2 (Biosafety Level 2) model viruses. Although the bacteriophage surrogate, Ø6, has been used to assess low pH BSL2 VI, it has not been used for evaluation of detergent-mediated VI. Furthermore, Ø6 is typically assayed through host cell infectivity which introduces the risk of cross-contaminating other cell lines in the facility. To circumvent contamination, we developed an in-house RT-qPCR (reverse transcriptase quantitative polymerase chain reaction) assay for selective detection of active Ø6 from a population of live and dead phage. The RT-qPCR assay was used to evaluate Ø6 inactivation in cell culture fluid of monoclonal antibody and fusion protein. Complementary Ø6 infectivity was also conducted at a third-party testing facility. The Ø6 RT-qPCR and infectivity data was modeled against VI of three BSL2 viruses, X- MuLV, A- MuLV and HSV-1 in corresponding therapeutics. Both Ø6 methods demonstrate that any VI agent showing Ø6 clearance of ≥ 2.5 logs would demonstrate complete BSL2 VI of ≥ 4.0 logs. Compared to BSL2 virus testing, this in-house Ø6 RT-qPCR tool can screen VI agents at 5% the cost and a turnaround time of 2-3 days versus 4-7 months.

A fundamental aspect of evolutionary biology is natural selection on trait variation. Classically, selection has been estimated primarily on external morphological traits such as beak size and coloration, or on easily-assayable physiological traits such as heat-tolerance. As technologies and methods improved, evolutionary biologists began examining selection on molecular traits such as protein sequences and cellular processes. In a From the Cover manuscript in this issue of Molecular Ecology, Ahmad et al. (2021) continue this trend by estimating parasite driven selection on the molecular trait of transcript abundance in a wild population of brown trout (Salmo trutta) by uniquely combining a mark-recapture experimental design with non-invasive RNA sampling. Using transcript abundance to estimate selection allows for many different traits (each unique gene’s transcript counts) to be tested in a single experiment, providing the opportunity to examine trends in selection. Ahmad et al.(2021) find directional selection strength on transcript counts is generally low and normally distributed. Surprisingly, transcripts under non-linear selection showed a disruptive selection bias contradicting previous comparative studies and theoretical work. This highlights the importance of within-generation selection studies, where mechanisms may differ from longer time frames. Their manuscript also highlights the benefits of an improved 3’ RNA sequencing technique to measure gene expression.

Predicting the fate of a microbial population (i.e., growth, gene expression…) remains a challenge, especially when this population is exposed to very dynamic environmental conditions, such as those encountered during continuous cultivation. Indeed, the dynamic nature of continuous cultivation process implies the potential deviation of the microbial population involving genotypic and phenotypic diversification. This work has been focused on the induction of the arabinose operon in Escherichia coli as a model system. As a preliminary step, the GFP level triggered by an arabinose-inducible ParaBAD promoter has been tracked by flow cytometry in chemostat with glucose-arabinose co-feeding. For a large range of glucose-arabinose co-feeding, the simultaneous occurrence of GFP positive and negative subpopulation was observed. In a second set of experiments, continuous cultivation was performed by adding either glucose or arabinose, based on the ability of individual cells for switching from low GFP to high GFP states, according to a technology called segregostat. In segregostat mode of cultivation, on-line flow cytometry analysis was used for adjusting the arabinose/glucose transitions based on the phenotypic switching capabilities of the microbial population. This strategy allowed finding an appropriate arabinose pulsing frequency, leading to a prolonged maintenance of the induction level with limited impact on phenotypic diversity for more than 60 generations. This result suggests that constraining individual cells into a given phenotypic trajectory is maybe not the best strategy for directing cell population. Instead, allowing individual cells switching around a predefined threshold seems to be a robust strategy leading to oscillating, but predictable, cell population behavior.

Objectives: Pneumothorax (PTX) in newborns is a life-threatening condition associated with high morbidity and mortality especially in premature infants. The frequency of PTX in neonates at different gestational ages (GA) and its impact on neonatal mortality have not been quantified. We aimed to determine: 1) the prevalence of PTX in neonates at different GA from ≤24 weeks to ≥37 weeks, 2) the impact of PTX on mortality per GA, and 3) the impact of PTX on the length of stay (LOS) per GA. Methods: The national Kids’ Inpatient Database (KID) for the years of 2006 to 2012 were used. We included all infants admitted to the hospital with a documented GA and ICD9 code of pneumothorax. Bivariate and multivariate analyses were conducted and odds ratios (OR) were calculated. Results: A total of 10 625 036 infants were included; of them 3665 infants (0.034 %) had a diagnosis of PTX, with highest prevalence at ≤24 weeks GA (0.67%), and lowest at term (0.02%). The overall mortality rate of patients with PTX was 8.8%, and greater in preterm (16.3%) vs. term infants (2.7%). The association of mortality with PTX was greatest at GA of 29−32 weeks (OR = 8.55 (95% CI: 6.56−11.13). Infants who survived until discharge had a median of 2–12 days longer length of stay depending on GA category. Conclusions: The prevalence of PTX peaks in infants <24 weeks, however its impact on mortality is greatest at 29-32 weeks. PTX is associated with longer length of stay in survivors.

Guillain-Barré syndrome (GBS) after surgery may be more common than previously recognized; however, GBS after open heart surgery is exceedingly rare. We describe a rare case of GBS occurring after aortic, tricuspid and mitral valve surgery and review the world’s literature. Vigilance for GBS among post-surgical patients is needed.

The case of a 45 year old female with chronic migraine undergoing treatment with erenumab 70 mg subcutaneous injection who developed Raynaud’s phenomenon (RP) as a side effect with relevant clinical implications two weeks after the second dose of erenumab injection.

Light pollution modelling and monitoring has traditionally used zenith sky brightness as its main indicator. Several other indicators (e.g. average sky radiance, horizontal irradiance, average sky radiance at given interval of zenith distances) may be more useful, both for general and for specific purposes of ecology studies, night sky and environmental monitoring. These indicators can be calculated after the whole sky radiance is known with sufficient angular detail. This means, for each site, to integrate the contribution in each direction of the sky of each light source in the radius of hundreds of km. This approach is extremely high time consuming if the mapping is desired for a large territory. Here we present a way to obtain maps of large territories for a large subset of useful indicators, bypassing the need to calculate first the radiance map of the whole sky in each site to obtain from it the desired indicator in that site. For each indicator, a point spread function (PSF) is calculated from the whole sky radiance maps generated by a single source at sufficiently dense number of distances from the observing site. If the PSF is transversally shift-invariant, i.e. if it depends only on the relative position of source and observer, then we can further speed up the map calculation via the use of fast Fourier-transform (FFT). We present here examples of maps for different indicators. Precise results can be calculated for any single site, taking into account the site and light sources altitudes, by means of specific inhomogeneous (spatially-variant) and anisotropic (non rotationally symmetric) PSFs.

The aim of this paper is to give a precise proof of the completeness of Lamb modes and associated modes. This proof is relatively simple and short but relies on two powerful mathematical theorems. The first one is a theorem on elliptic systems with a parameter due to Agranovich and Vishik. The second one is a theorem due to Locker which gives a criterion to show the completeness of the set of generalized eigenvectors of a Hilbert-Schmidt discrete operator.

1. Restoration ecology has historically focused on reconstructing communities of highly visible taxa whilst less visible taxa, such as invertebrates and microbes, are ignored. This is problematic as invertebrates and microbes make up the vast bulk of biodiversity and drive many key ecosystem processes, yet they are rarely actively reintroduced following restoration, potentially limiting ecosystem function and biodiversity in these areas. 2. In this review, we discuss the current (limited) incorporation of invertebrates and microbes in restoration and rewilding projects. We argue that these groups should be actively rewilded during restoration to improve biodiversity and ecosystem function outcomes and highlight how they can be used to greater effect in the future. For example, invertebrates and microbes are easily manipulated, meaning whole communities can potentially be rewilded through habitat transplants in a practice that we refer to as “whole-of-community” rewilding. 3. We provide a framework for whole-of-community rewilding and describe empirical case studies as practical applications of this under-researched restoration tool that land managers can use to improve restoration outcomes. 4. We hope this new perspective on whole-of-community restoration will promote applied research into restoration that incorporates all biota, irrespective of size, whilst also enabling a better understanding of fundamental ecological theory, such as colonisation- competition trade-offs. This may be a necessary consideration as invertebrates that are important in providing ecosystem services are declining globally; targeting invertebrate communities during restoration may be crucial in stemming this decline.

Marine Lenhart Syndrome (MLS) is caused by a coexistence of active thyroid nodules and Graves’ disease1. Here, we present a case of hyperthyroidism characterized by the presence of stimulating TSH receptor antibodies, unsuccessful radioactive iodine ablation, ultimately requiring Methimazole followed by thyroidectomy. We review the current literature.

Species delimitation among closely related species is challenging because traditional phenotype-based approaches, e.g., morphology, ecological, or chemical characteristics, often produce conflicting results. With the advent of high-throughput sequencing, it has become increasingly cost-effective to acquire genome-scale data which can resolve previously ambiguous species boundaries. As the availability of genome-scale data has increased, numerous species delimitation analyses, such as BPP and SNAPP+Bayes factor delimitation (BFD*), have been developed to delimit species boundaries. However, even empirical molecular species delimitation approaches can be biased by confounding evolutionary factors, e.g., hybridization/introgression and incomplete lineage sorting, and computational limitations. Here we investigate species boundaries and the potential for micro-endemism in a lineage of lichen-forming fungi, Niebla Rundel & Bowler in the family Ramalinaceae. The species delimitation models tend to support more specious groupings, but were unable to infer robust, consistent species delimitations. The results of our study highlight the problem of delimiting species, particularly in groups such as Niebla, with complex, recent phylogeographic histories.

Appendiceal diseases are relatively rare reported complications during hematopoietic stem cell transplantation with no guidance on management. Pre- and post-transplant patients should receive a trial of medical therapy with appendectomy after recovery but prior to transplant in the former and plan for appendectomy after completion of immunosuppression in the latter.

Xq28 (involving MECP2) duplication syndrome is a severe neurodevelopmental disorder in males, most females are asymptomatic carriers, but there are phenotypic heterogeneities in the females. Skewed X-chromosome inactivation (XCI) seems to prevent duplicated region activation in asymptomatic females, but it remains controversial. Herein we reported two asymptomatic females (daughter and mother) with interstitial Xq28 duplication. HUMARA and RP2 assays showed that both had complete skewed XCI, the Xq28 duplicated chromosome was inactivated in the daughter, but surprisingly, it was activated in her mother. Interestingly, by combining RNA sequencing and whole-exome sequencing, we confirmed that XIST only expressed in the Xq28 duplication chromosomes of the two females, indicating that the Xq28 duplication chromosomes were inactive. Meanwhile, MECP2 and most XCI genes in the duplicated X-chromosomes were not transcriptionally expressed or upregulated, precluding major clinical phenotypes in the two females, especially the mother. We showed that XCI status detected by RNA sequencing was more relevant for establishing the clinical phenotype of MECP2 duplication females. It suggested there were other factors maintaining the XCI status in addition to DNA methylation, a possible additional inhibition mechanism occured at the transcriptional level in the unmethylated X-chromosome, counter balancing the MECP2 duplication’s detrimental phenotype effects

Letter to the Editor, BJOG Title:Deceleration Area and Deceleration Capacity: Promising predictors of fetal acidaemia in human labour? Visual versus computerised cardiotocographyRe: Georgieva A, Lear CA, Westgate JA, Kasai M, Miyagi E, Ikeda T, Gunn AJ, Bennet L. Deceleration area and capacity during labour-like umbilical cord occlusions identify evolving hypotension: a controlled study in fetal sheep. BJOG 2021; https://doi.org/10.1111/1471-0528.16638.Author: Mr. Shashikant L SHOLAPURKARMD, DNB, MRCOGDept of Obstetrics & Gynaecology,Royal United Hospital, Bath, BA1 3NG, UKEmail:[email protected]; [email protected]; Tel: 07906620662Short Running Title: Deceleration area and capacity in labourWord count: 500Corresponding Author: Mr. Shashikant L SHOLAPURKARMD, DNB, MRCOGDept of Obstetrics & Gynaecology,Royal United Hospital, Bath, BA1 3NG, UKStatement of interest: The author has no conflict of interest or funding to declare.

In this paper we consider the propagation of waves in an open waveguide in R^2 where the index of refraction is a local perturbation of a function which is periodic along the axis of the waveguide and equal to one outside a strip of finite width. Motivated by the limiting absorption principle (proven in an ealier paper by the author for the case of an open waveguide in the half space) we formulate a radiation condition which allows the existence of propagating modes and prove uniqueness, existence, and stability of a solution. In the last part we investigate the decay properties of the radiating part in the direction of periodicity and orthogonal to it.

Humans and non-human primates (NHPs) harbor complex gut microbial communities that affect phenotypes and fitness. The gut microbiotas of wild NHPs reflect their hosts’ phylogenetic histories and are compositionally distinct from those of humans, but in captivity the endogenous gut microbial lineages of NHPs can be lost or replaced by lineages found in humans. Despite its potential contributions to gastrointestinal dysfunction, this humanization of the gut microbiota has not been investigated systematically across captive NHP species. Here we show through comparisons of well-sampled wild and captive populations of apes and monkeys that the fraction of the gut microbiota humanized by captivity varies significantly between NHP species but is remarkably reproducible between captive populations of the same NHP species. Conspecific captive populations displayed significantly greater than expected overlap in the sets of bacterial 16S rRNA gene variants that were differentially abundant between captivity and the wild. This overlap was evident even between captive populations residing on different continents but was never observed between heterospecific captive populations. In addition, we developed an approach incorporating human gut microbiota data to rank NHPs’ gut microbial clades based on the propensity of their lineages to be lost or replaced by lineages found in humans in captivity. Relatively few microbial genera displayed reproducible degrees of humanization in different captive host species, but most microbial genera were reproducibly humanized or retained from the wild in conspecific pairs of captive populations. These results demonstrate that the gut microbiotas of captive NHPs display predictable, host-species specific responses to captivity.

Resource availability and heterogeneity are recognized as two essential environmental aspects to determine species diversity and community abundance. However, how resource availability and heterogeneity determine species diversity and community abundance in highly heterogeneous and most fragile karst landscapes is largely unknown. We examined the effects of resource availability and heterogeneity on plant community composition and quantified their relative contribution by variation partitioning. Then, a structural equation model (SEM) was used to further disentangle the multiple direct and indirect effects of resource availability on plant community composition. Species diversity was significantly influenced by the resource availability in shrubland and woodland but not by the heterogeneity in woodland. Abundance was significantly affected by both resource availability and heterogeneity, whereas variation partitioning results showed that resource availability explained the majority of the variance in abundance, and the contribution of resource heterogeneity was marginal. These results indicated that resource availability plays a more important role in determining karst plant community composition than resource heterogeneity. Our SEMs further found that the multiple direct and indirect processes of resource availability in determining karst species diversity and abundance were different in different vegetation types. Resource availability and heterogeneity both played a certain role in determining karst plant community composition, while the importance of resource availability far exceeded resource heterogeneity. We propose that steering community restoration and reconstruction should be highly dependent on resource availability, and multiple direct and indirect pathways of resource availability for structuring karst plant communities need to be taken into account.

Objectives: Our study aimed to identify potential factors that influence rehabilitation outcomes in late-implanted adolescents and adults with prelingual deafness and attempted to develop a reliable nomogram. Design: This retrospective study included 120 subjects less than 30 years of age who had received cochlear implantation at a single medical center. The Categories of Auditory Performance (CAP) scale was used to evaluate the rehabilitation outcomes. A nomogram was constructed using the R and EmpowerStats software. Results: Univariate analysis indicated higher rates of auditory performance improvement in younger aged subjects. Residual hearing and regular and longer implant use were more common among subjects showing auditory performance improvement. Multivariate analysis identified residual hearing (Hazard Ratio, 6.11; 95% Confidence Interval, 1.83-20.41; P<0.01), age at implantation (Hazard Ratio, 0.31; 95% Confidence Interval, 0.14-0.83; P=0.02) and regular CI use (Hazard Ratio, 7.79; 95% Confidence Interval, 2.50-24.20; P<0.01) as independent predictors for auditory performance improvement. The nomogram’s predictive performance was satisfactory as verified by the calibration curve and Receiver operating characteristic (ROC) curve. Conclusions: We demonstrated that residual hearing, younger age, and regular CI use were associated with auditory performance improvement in this cochlear implant user population. Our nomogram model demonstrated satisfactory predictive performance for this population.