Spatial Multiplexing of Fluorescent Reporters for Imaging Signaling Network Dynamics.

In order to analyze how a signal transduction network converts cellular inputs into cellular outputs, ideally one would measure the dynamics of many signals within the network simultaneously. We found that, by fusing a fluorescent reporter to a pair of self-assembling peptides, it could be stably clustered within cells at random points, distant enough to be resolved by a microscope but close enough to spatially sample the relevant biology. Because such clusters, which we call signaling reporter islands (SiRIs), can be modularly designed, they permit a set of fluorescent reporters to be efficiently adapted for simultaneous measurement of multiple nodes of a signal transduction network within single cells. We created SiRIs for indicators of second messengers and kinases and used them, in hippocampal neurons in culture and intact brain slices, to discover relationships between the speed of calcium signaling, and the amplitude of PKA signaling, upon receiving a cAMP-driving stimulus.

Serial Interval and Incubation Period Estimates of Monkeypox Virus Infection in 12 Jurisdictions, United States, May-August 2022.

Using data from 12 US health departments, we estimated mean serial interval for monkeypox virus infection to be 8.5 (95% credible interval 7.3-9.9) days for symptom onset, based on 57 case pairs. Mean estimated incubation period was 5.6 (95% credible interval 4.3-7.8) days for symptom onset, based on 35 case pairs.

Cryptosporulation in Kurthia spp. forces a rethinking of asporogenesis in Firmicutes.

Endosporulation is a complex morphophysiological process resulting in a more resistant cellular structure that is produced within the mother cell and is called endospore. Endosporulation evolved in the common ancestor of Firmicutes, but it is lost in descendant lineages classified as asporogenic. While Kurthia spp. is considered to comprise only asporogenic species, we show here that strain 11kri321, which was isolated from an oligotrophic geothermal reservoir, produces phase-bright spore-like structures. Phylogenomics of strain 11kri321 and other Kurthia strains reveals little similarity to genetic determinants of sporulation known from endosporulating Bacilli. However, morphological hallmarks of endosporulation were observed in two of the four Kurthia strains tested, resulting in spore-like structures (cryptospores). In contrast to classic endospores, these cryptospores did not protect against heat or UV damage and successive sub-culturing led to the loss of the cryptosporulating phenotype. Our findings imply that a cryptosporulation phenotype may have been prevalent and subsequently lost by laboratory culturing in other Firmicutes currently considered as asporogenic. Cryptosporulation might thus represent an ancestral but unstable and adaptive developmental state in Firmicutes that is under selection under harsh environmental conditions.

Evolthon: A community endeavor to evolve lab evolution.

In experimental evolution, scientists evolve organisms in the lab, typically by challenging them to new environmental conditions. How best to evolve a desired trait? Should the challenge be applied abruptly, gradually, periodically, sporadically? Should one apply chemical mutagenesis, and do strains with high innate mutation rate evolve faster? What are ideal population sizes of evolving populations? There are endless strategies, beyond those that can be exposed by individual labs. We therefore arranged a community challenge, Evolthon, in which students and scientists from different labs were asked to evolve Escherichia coli or Saccharomyces cerevisiae for an abiotic stress-low temperature. About 30 participants from around the world explored diverse environmental and genetic regimes of evolution. After a period of evolution in each lab, all strains of each species were competed with one another. In yeast, the most successful strategies were those that used mating, underscoring the importance of sex in evolution. In bacteria, the fittest strain used a strategy based on exploration of different mutation rates. Different strategies displayed variable levels of performance and stability across additional challenges and conditions. This study therefore uncovers principles of effective experimental evolutionary regimens and might prove useful also for biotechnological developments of new strains and for understanding natural strategies in evolutionary arms races between species. Evolthon constitutes a model for community-based scientific exploration that encourages creativity and cooperation.

Epidemiologic and Clinical Characteristics of Monkeypox Cases - United States, May 17-July 22, 2022.

Monkeypox, a zoonotic infection caused by an orthopoxvirus, is endemic in parts of Africa. On August 4, 2022, the U.S. Department of Health and Human Services declared the U.S. monkeypox outbreak, which began on May 17, to be a public health emergency (1,2). After detection of the first U.S. monkeypox case), CDC and health departments implemented enhanced monkeypox case detection and reporting. Among 2,891 cases reported in the United States through July 22 by 43 states, Puerto Rico, and the District of Columbia (DC), CDC received case report forms for 1,195 (41%) cases by July 27. Among these, 99% of cases were among men; among men with available information, 94% reported male-to-male sexual or close intimate contact during the 3 weeks before symptom onset. Among the 88% of cases with available data, 41% were among non-Hispanic White (White) persons, 28% among Hispanic or Latino (Hispanic) persons, and 26% among non-Hispanic Black or African American (Black) persons. Forty-two percent of persons with monkeypox with available data did not report the typical prodrome as their first symptom, and 46% reported one or more genital lesions during their illness; 41% had HIV infection. Data suggest that widespread community transmission of monkeypox has disproportionately affected gay, bisexual, and other men who have sex with men and racial and ethnic minority groups. Compared with historical reports of monkeypox in areas with endemic disease, currently reported outbreak-associated cases are less likely to have a prodrome and more likely to have genital involvement. CDC and other federal, state, and local agencies have implemented response efforts to expand testing, treatment, and vaccination. Public health efforts should prioritize gay, bisexual, and other men who have sex with men, who are currently disproportionately affected, for prevention and testing, while addressing equity, minimizing stigma, and maintaining vigilance for transmission in other populations. Clinicians should test patients with rash consistent with monkeypox,† regardless of whether the rash is disseminated or was preceded by prodrome. Likewise, although most cases to date have occurred among gay, bisexual, and other men who have sex with men, any patient with rash consistent with monkeypox should be considered for testing. CDC is continually evaluating new evidence and tailoring response strategies as information on changing case demographics, clinical characteristics, transmission, and vaccine effectiveness become available.§.

Allopatric and Sympatric Drivers of Speciation in Alviniconcha Hydrothermal Vent Snails.

Despite significant advances in our understanding of speciation in the marine environment, the mechanisms underlying evolutionary diversification in deep-sea habitats remain poorly investigated. Here, we used multigene molecular clocks and population genetic inferences to examine processes that led to the emergence of the six extant lineages of Alviniconcha snails, a key taxon inhabiting deep-sea hydrothermal vents in the Indo-Pacific Ocean. We show that both allopatric divergence through historical vicariance and ecological isolation due to niche segregation contributed to speciation in this genus. The split between the two major Alviniconcha clades (separating A. boucheti and A. marisindica from A. kojimai, A. hessleri, and A. strummeri) probably resulted from tectonic processes leading to geographic separation, whereas the splits between co-occurring species might have been influenced by ecological factors, such as the availability of specific chemosynthetic symbionts. Phylogenetic origin of the sixth species, Alviniconcha adamantis, remains uncertain, although its sister position to other extant Alviniconcha lineages indicates a possible ancestral relationship. This study lays a foundation for future genomic studies aimed at deciphering the roles of local adaptation, reproductive biology, and host-symbiont compatibility in speciation of these vent-restricted snails.

A thematic analysis of in-hospital end-of-life care experiences of surviving family members.

While end-of-life care (EoLC) priorities for patients dying in the hospital are well-documented, few data characterize needs and experiences of their family members. We conducted thematic analysis of audio recorded interviews of 18 bereaved family members to elucidate these experiences. Participants' memories were organized into two parent themes: those related to satisfaction with the care received and effective communication; those identifying shortcomings in patient care, hospital-family communication, hospital environment, and care burden on the part of family members. These findings provide insight to enhance services to patients and their families at end-of-life and improve postmortem and bereavement services.

Indoor nature exposure and influence on physiological stress markers.

Spending time in nature is beneficial for stress reduction and recovery. Using the properties of biophilic design, this study examined the influence of a nature-based indoor environment on physiological stress systems. An experimental study was designed to assess the influence of indoor natural elements on autonomic activity (heart rate variability or HRV), self-reported environmental assessments. No differences in heart rate variability were found between participants assigned to either condition. The room with natural elements was rated more positively than the room without natural elements. Participant preference had more impact on changes in HRV for participants without exposure to natural elements. The results suggest that natural elements in indoor environments may influence the regulation of stress response via environmental preference.Abbreviations: ANCOVA: Analysis of Covariance; ANS: Autonomic Nervous System; AVNN: Average of NN; DST: Digit Span Test; EAS: Environmental Assessment Scale; ECG: Electrocardiograph; fMRI: Functional magnetic resonance imaging; HF: High Frequency; HRV: Heart Rate Variability; INE: Indoor Nature Exposure; NR: Nature-Relatedness Scale; PANAS: Positive and Negative Affect Schedule; SAM: Search and Memory Test; SLSI: Student Life Stress Inventory; SMT: Sentence Making Test.

Use of Spirituality to Make Sense of Loss by Homicide.

This article presents a magnification of Stage 2 of the Theory of Post-Homicide Spiritual Change, a three-stage grounded theory of spiritual change after homicide (Theory of PHSC). Having endured the disintegration of their belief systems in the immediate aftermath of murder (Stage 1), survivors turn in Stage 2 to a more extended process of grappling with a crisis of meaning. This Stage 2 process is presented within the framework of the meaning making model, with attention to spiritual meaning making and transcendental experiences. Findings can help service providers support homicide survivors throughout an intermediary stage of bereavement that is marked by a sense of stagnation and diminished well-being. By accompanying survivors through the difficult meaning making efforts that characterize this stage, providers can help position them to break free of intensive cognitive meaning making and gain forward momentum in Stage 3 of the Theory of PHSC and can focus on aspects of life that can help them successfully make meaning of their loss while positioning them to gain forward momentum.

Evaluating nursing faculty's approach to information literacy instruction: a multi-institutional study.

OBJECTIVE: In 2018, the Association of College & Research Libraries (ACRL) Health Sciences Interest Group convened a working group to update the 2013 Information Literacy Competency Standards for Nursing to be a companion document to the 2016 Framework for Information Literacy in Higher Education. To create this companion document, the working group first needed to understand how nursing faculty approached information literacy (IL) instruction. METHODS: The working group designed a survey that assessed how nursing faculty utilized IL principles in coursework and instruction. The survey consisted of nineteen mixed methods questions and was distributed to nursing faculty at eight institutions across the United States. RESULTS: Most (79%) faculty indicated that they use a variety of methods to teach IL principles in their courses. While only 12% of faculty incorporated a version of the ACRL IL competencies in course design, they were much more likely to integrate nursing educational association standards. Faculty perceptions of the relevance of IL skills increased as the education level being taught increased. CONCLUSION: The integration of IL instruction into nursing education has mostly been achieved through using standards from nursing educational associations. Understanding these standards and understanding how faculty perceptions of the relevance of IL skills change with educational levels will guide the development of a companion document that librarians can use to collaborate with nurse educators to integrate IL instruction throughout nursing curriculums at course and program levels.

A Grounded Theory of the Process of Spiritual Change Among Homicide Survivors.

Grounded theory was used to generate a mid-range theory of the process of spiritual change in the lives of survivors of homicide victims. Theoretical sampling guided the selection of 30 participants from a larger study of spiritual change after homicide (N = 112). Individual interviews were analyzed using a four-step sequence of line-by-line, focused, axial, and selective coding. Analysis generated a closed theory consisting of three fluids, consecutive but nonlinear stages. Each stage consisted of an overarching process and a state of being in the world: (a) Disintegrating: living in a state of shock; (b) Reckoning: living in a state of stagnation; (c) Recreating and reintegrating the self: living in a state of renewal. Movement through the stages was fueled by processes of spiritual connection that yielded changes that permeated the theory. Findings can be used to help practitioners address the processes that drive spiritual change in the lives of homicide survivors.

Host hybridization as a potential mechanism of lateral symbiont transfer in deep-sea vesicomyid clams.

Deep-sea vesicomyid clams live in mutualistic symbiosis with chemosynthetic bacteria that are inherited through the maternal germ line. On evolutionary timescales, strictly vertical transmission should lead to cospeciation of host mitochondrial and symbiont lineages; nonetheless, examples of incongruent phylogenies have been reported, suggesting that symbionts are occasionally horizontally transmitted between host species. The current paradigm for vesicomyid clams holds that direct transfers cause host shifts or mixtures of symbionts. An alternative hypothesis suggests that hybridization between host species might explain symbiont transfers. Two clam species, Archivesica gigas and Phreagena soyoae, frequently co-occur at deep-sea hydrocarbon seeps in the eastern Pacific Ocean. Although the two species typically host gammaproteobacterial symbiont lineages marked by divergent 16S rRNA phylotypes, we identified a number of clams with the A. gigas mitotype that hosted symbionts with the P. soyoae phylotype. Demographic inference models based on genome-wide SNP data and three Sanger sequenced gene markers provided evidence that A. gigas and P. soyoae hybridized in the past, supporting the hypothesis that hybridization might be a viable mechanism of interspecific symbiont transfer. These findings provide new perspectives on the evolution of vertically transmitted symbionts and their hosts in deep-sea chemosynthetic environments.

Laser depolarization ratio measurement of corn leaves from the biochar and non-biochar applied plots.

Research has shown that biochar, when used as a soil amendment, can improve soil quality and potentially increase agriculture production. This case study was targeted at comparing the polarimetric laser reflectivities of the corn (Zea mays L.) leaf samples collected from two plots: the biochar plot with corn stover biochar previously applied, and the control plot without any biochar treatment. Specifically, measurements of the co-polarization and the cross-polarization components of the corn leaves at 532-nm laser wavelength were performed. It was discovered that the leaf samples collected from the control plot had larger depolarization ratio in comparison to these from the biochar plot. Data analysis showed that the depolarization difference was statistically significant. Such difference is attributed to the application of biochar. This study agrees with the conclusion that biochar improves soil fertility from previous literature using direct measures. In addition, it suggests that laser depolarization ratio can be used as an indicator to monitor plant growth condition. Furthermore, our finding improves the understanding and application of leaf laser polarimetry, provides future research directions toward the leaf and plant polarimetric scattering models, and will contribute to the design of future remote sensing polarimetric lidar in agriculture and forest remote sensing.

A Hermeneutic Phenomenological Study of the Lived Experience of Spiritual Conversion in a Neo-Charismatic Evangelical Context.

Hermeneutic phenomenology was used to illuminate the lived experience of posttraumatic, born-again conversion among members of a Neo-Charismatic Evangelical church (n = 11). Data were analyzed using a six-step process that yielded six core constituents: (1) Catalysts for transformation; (2) Courses of spiritual conversion; (3) Relationship with God; (4) Receiving the love of God; (5) Transformed by the love of God; (6) Changes in relationships. The essence of the phenomenon was confident knowing. Methodological rigor was maximized via maintenance of an audit trail and memo-writing. Findings highlight the essence of a spiritual conversion experience that is central to the faith and lives of many people, but remains poorly understood. The insights provided can build understanding of conversion among mental health providers, thereby reducing avoidance of the topic and promoting informed assistance for individuals experiencing a conversion process.

Hydrothermal vent fields discovered in the southern Gulf of California clarify role of habitat in augmenting regional diversity.

Hydrothermal vent communities are distributed along mid-ocean spreading ridges as isolated patches. While distance is a key factor influencing connectivity among sites, habitat characteristics are also critical. The Pescadero Basin (PB) and Alarcón Rise (AR) vent fields, recently discovered in the southern Gulf of California, are bounded by previously known vent localities (e.g. Guaymas Basin and 21° N East Pacific Rise); yet, the newly discovered vents differ markedly in substrata and vent fluid attributes. Out of 116 macrofaunal species observed or collected, only three species are shared among all four vent fields, while 73 occur at only one locality. Foundation species at basalt-hosted sulfide chimneys on the AR differ from the functional equivalents inhabiting sediment-hosted carbonate chimneys in the PB, only 75 km away. The dominant species of symbiont-hosting tubeworms and clams, and peripheral suspension-feeding taxa, differ between the sites. Notably, the PB vents host a limited and specialized fauna in which 17 of 26 species are unknown at other regional vents and many are new species. Rare sightings and captured larvae of the 'missing' species revealed that dispersal limitation is not responsible for differences in community composition at the neighbouring vent localities. Instead, larval recruitment-limiting habitat suitability probably favours species differentially. As scenarios develop to design conservation strategies around mining of seafloor sulfide deposits, these results illustrate that models encompassing habitat characteristics are needed to predict metacommunity structure.

Reliability and Validity of the Sexual Experiences Survey-Short Forms Victimization and Perpetration.

The Sexual Experiences Survey (SES), the most widely used measure of unwanted sexual experiences, was recently updated (Koss et al., 2007). The purpose of this study was to provide psychometric data on the updated Sexual Experiences Survey-Short Form Perpetration (SES-SFP) and the Sexual Experiences Survey-Short Form Victimization (SES-SFV). Men (n = 136) and women (n = 433) were randomly assigned to in-person or Internet formats of administration for 3 measurement points. Women completed victimization surveys and trauma measures. Men completed perpetration surveys and attitude/ personality measures. Results supported the validity and reliability of both the SES-SFV with women and the SES-SFP with men. Further research is needed regarding the use of the SES-SFV with men and the SES-SFP with women.

Presumptive primary pulmonary mast cell tumor in 2 dogs.

Two dogs were presented, each with a large solitary pulmonary mass, and cytology confirmed mast cell tumor (MCT) in each dog. One dog was euthanized following diagnosis. Thoracic computed tomography scan and exploratory thoracotomy of the second dog revealed a right pulmonary mass that would require a radical lung resection. The patient was euthanized and histopathology confirmed a poorly granulated MCT with characteristics suggestive of epitheliotropism, an uncommon finding with MCT. These represent the first reported cases of presumptive primary pulmonary MCT in dogs.

Mastocytome primaire pulmonaire présumé chez deux chiens. Deux chiens ont été présentés avec une volumineuse masse pulmonaire dont l'analyse cytopathologique confirma le diagnostic de mastocytome (MCT). L'un des chiens a été euthanasié suite au diagnostic. Un CT scan thoracique et une thoracotomie exploratrice du second chien ont révélé une masse pulmonaire droite nécessitant une résection pulmonaire radicale; le chien fut euthanasié. L'histopathologie a confirmé un MCT peu granulé avec des caractéristiques suggestives d'épithéliotropisme, une trouvaille inhabituelle lors de MCT. Il s'agit des premiers cas rapportés de MCT pulmonaires primaires présumés chez le chien.(Traduit par les auteurs).

Population subdivision of hydrothermal vent polychaete Alvinella pompejana across equatorial and Easter Microplate boundaries.

BACKGROUND: The Equator and Easter Microplate regions of the eastern Pacific Ocean exhibit geomorphological and hydrological features that create barriers to dispersal for a number of animals associated with deep-sea hydrothermal vent habitats. This study examined effects of these boundaries on geographical subdivision of the vent polychaete Alvinella pompejana. DNA sequences from one mitochondrial and eleven nuclear genes were examined in samples collected from ten vent localities that comprise the species' known range from 23°N latitude on the East Pacific Rise to 38°S latitude on the Pacific Antarctic Ridge. RESULTS: Multi-locus genotypes inferred from these sequences clustered the individual worms into three metapopulation segments - the northern East Pacific Rise (NEPR), southern East Pacific Rise (SEPR), and northeastern Pacific Antarctic Ridge (PAR) - separated by the Equator and Easter Microplate boundaries. Genetic diversity estimators were negatively correlated with tectonic spreading rates. Application of the isolation-with-migration (IMa2) model provided information about divergence times and demographic parameters. The PAR and NEPR metapopulation segments were estimated to have split roughly 4.20 million years ago (Mya) (2.42-33.42 Mya, 95 % highest posterior density, (HPD)), followed by splitting of the SEPR and NEPR segments about 0.79 Mya (0.07-6.67 Mya, 95 % HPD). Estimates of gene flow between the neighboring regions were mostly low (2 Nm < 1). Estimates of effective population size decreased with southern latitudes: NEPR > SEPR > PAR. CONCLUSIONS: Highly effective dispersal capabilities allow A. pompejana to overcome the temporal instability and intermittent distribution of active hydrothermal vents in the eastern Pacific Ocean. Consequently, the species exhibits very high levels of genetic diversity compared with many co-distributed vent annelids and mollusks. Nonetheless, its levels of genetic diversity in partially isolated populations are inversely correlated with tectonic spreading rates. As for many other vent taxa, this pioneering colonizer is similarly affected by local rates of habitat turnover and by major dispersal filters associated with the Equator and the Easter Microplate region.

Nearly finished genomes produced using gel microdroplet culturing reveal substantial intraspecies genomic diversity within the human microbiome.

The majority of microbial genomic diversity remains unexplored. This is largely due to our inability to culture most microorganisms in isolation, which is a prerequisite for traditional genome sequencing. Single-cell sequencing has allowed researchers to circumvent this limitation. DNA is amplified directly from a single cell using the whole-genome amplification technique of multiple displacement amplification (MDA). However, MDA from a single chromosome copy suffers from amplification bias and a large loss of specificity from even very small amounts of DNA contamination, which makes assembling a genome difficult and completely finishing a genome impossible except in extraordinary circumstances. Gel microdrop cultivation allows culturing of a diverse microbial community and provides hundreds to thousands of genetically identical cells as input for an MDA reaction. We demonstrate the utility of this approach by comparing sequencing results of gel microdroplets and single cells following MDA. Bias is reduced in the MDA reaction and genome sequencing, and assembly is greatly improved when using gel microdroplets. We acquired multiple near-complete genomes for two bacterial species from human oral and stool microbiome samples. A significant amount of genome diversity, including single nucleotide polymorphisms and genome recombination, is discovered. Gel microdroplets offer a powerful and high-throughput technology for assembling whole genomes from complex samples and for probing the pan-genome of naturally occurring populations.