Dielectrophoretic bead-droplet reactor for solid-phase synthesis.

Solid-phase synthesis underpins many advances in synthetic and combinatorial chemistry, biology, and material science. The immobilization of a reacting species on the solid support makes interfacing of reagents an important challenge in this approach. In traditional synthesis columns, this leads to reaction errors that limit the product yield and necessitates excess consumption of the mobile reagent phase. Although droplet microfluidics can mitigate these problems, its adoption is fundamentally limited by the inability to controllably interface microbeads and reagent droplets. Here, we introduce Dielectrophoretic Bead-Droplet Reactor as a physical method to implement solid-phase synthesis on individual functionalized microbeads by encapsulating and ejecting them from microdroplets by tuning the supply voltage. Proof-of-concept demonstration of the enzymatic coupling of fluorescently labeled nucleotides onto the bead using this reactor yielded a 3.2-fold higher fidelity over columns through precise interfacing of individual microreactors and beads. Our work combines microparticle manipulation and droplet microfluidics to address a long-standing problem in solid-phase synthesis with potentially wide-ranging implications.

Apical Abscess on a Zygomaticus Implant Initially Diagnosed as a Cutaneous Carcinoma.

We present a case of an infection on a zygomaticus implant presenting on the skin, mimicking a cutaneous carcinoma, and presenting to a head and neck tumor board. The clinical findings were an intermittently discharging lesion over the zygomatic bone, which resolved upon removing the offending zygomaticus implant. It is essential to be aware that infections on a zygomaticus implant can occur well away from the normal tooth-bearing areas, and having a dentist with knowledge of these implants on a tumor board can prevent misdiagnosis and treatment.

Functional identification of bacterial spermine, thermospermine, norspermine, norspermidine, spermidine, and N1-aminopropylagmatine synthases.

Spermine synthase is an aminopropyltransferase that adds an aminopropyl group to the essential polyamine spermidine to form tetraamine spermine, needed for normal human neural development, plant salt and drought resistance, and yeast CoA biosynthesis. We functionally identify for the first time bacterial spermine synthases, derived from phyla Bacillota, Rhodothermota, Thermodesulfobacteriota, Nitrospirota, Deinococcota, and Pseudomonadota. We also identify bacterial aminopropyltransferases that synthesize the spermine same mass isomer thermospermine, from phyla Cyanobacteriota, Thermodesulfobacteriota, Nitrospirota, Dictyoglomota, Armatimonadota, and Pseudomonadota, including the human opportunistic pathogen Pseudomonas aeruginosa. Most of these bacterial synthases were capable of synthesizing spermine or thermospermine from the diamine putrescine and so possess also spermidine synthase activity. We found that most thermospermine synthases could synthesize tetraamine norspermine from triamine norspermidine, that is, they are potential norspermine synthases. This finding could explain the enigmatic source of norspermine in bacteria. Some of the thermospermine synthases could synthesize norspermidine from diamine 1,3-diaminopropane, demonstrating that they are potential norspermidine synthases. Of 18 bacterial spermidine synthases identified, 17 were able to aminopropylate agmatine to form N1-aminopropylagmatine, including the spermidine synthase of Bacillus subtilis, a species known to be devoid of putrescine. This suggests that the N1-aminopropylagmatine pathway for spermidine biosynthesis, which bypasses putrescine, may be far more widespread than realized and may be the default pathway for spermidine biosynthesis in species encoding L-arginine decarboxylase for agmatine production. Some thermospermine synthases were able to aminopropylate N1-aminopropylagmatine to form N12-guanidinothermospermine. Our study reveals an unsuspected diversification of bacterial polyamine biosynthesis and suggests a more prominent role for agmatine.

Sticker-Type Remote Monitoring System for Early Risk Detection of Catheter Associated Urinary Tract Infections.

A substantial number of critically ill patients in intensive care units (ICUs) rely on indwelling urinary catheters (IDCs), demanding regular monitoring of urine bags. This process increases the workload for healthcare providers and elevates the risk of exposure to contagious diseases. Moreover, IDCs are a primary cause of catheter-associated urinary tract infections (UTIs) in ICU patients whose delayed detection can have life-threatening complications. To address this, we have developed a Sticker Type Antenna for Remote Sensing (STARS) system capable of measuring urine flow rate and conductivity as early-risk markers for UTIs, alongside tracking patients' urine bag status to facilitate medical automation for healthcare providers. STARS comprises a simple, low-cost, disposable antenna module for contactless measurements of urine volume and conductivity, and a reusable wireless module for real-time data transmission. Systematic studies on STARS revealed its stable performance within physiologically relevant ranges of urine volume (0 to 2000 ml) and conductivity (5 to 40 mS/cm) in urine bags. As a proof-of-concept, STARS was tested in artificially created healthy and infected urine specimens to validate its non-contact sensing performance in detecting the onset of UTIs in catheterized patients within a hospital-like environment. STARS represents the first application of a real-time, contactless, wireless monitoring platform for simultaneous urine bag management and early risk detection of UTIs.

Could nudges reduce health literacy disparities in CVD prevention? An experiment using alternative messages for CVD risk assessment screening.

OBJECTIVE: To explore the effect of SMS nudge messages amongst people with varying health literacy on their intention to get a Heart Health Check. METHODS: A 3 (Initial SMS: scarcity, regret, or control nudge) x 2 (Reminder SMS: social norm or control nudge) factorial design was used in a hypothetical online experiment. 705 participants eligible for Heart Health Checks were recruited. Outcomes included intention to attend a Heart Health Check and psychological responses. RESULTS: In the control condition, people with lower health literacy had lower behavioural intentions compared to those with higher health literacy (p = .011). Scarcity and regret nudges closed this gap, resulting in similar intention levels for lower and higher health literacy. There was no interactive effect of the reminder nudge and health literacy (p = .724). CONCLUSION: Scarcity and regret nudge messages closed the health literacy gap in behavioural intentions compared to a control message, while a reminder nudge had limited additional benefit. Health literacy should be considered in behavioural intervention evaluations to ensure health equity is addressed. PRACTICE IMPLICATIONS: Results informed a national screening program using a universal precautions approach, where messages with higher engagement for lower health literacy groups were used in clinical practice.

Rosa Bonheur the Amazon? Equestrianism, female masculinity, and The Horse Fair (1852-1855).

In 1853, Rosa Bonheur first exhibited what would become her most widely celebrated work: The Horse Fair. Although the work's modern setting and animal-focused subject matter do not obviously characterize it as an instance of classical reception, the artist claimed that it was inspired by the Parthenon frieze. A significant amount of feminist and queer scholarship has been dedicated to Rosa Bonheur's life, career, and art practices, all of which reveal the complex ways in which the artist negotiated the gender norms of 19th-century France. These ranged from her decision never to marry, instead living in households with two women, to her officially sanctioned practice of cross-dressing when conducting art studies in public. In view of all these things, one of the most remarkable elements of The Horse Fair is the very probable inclusion of the artist's self-portrait, clad in masculine clothing and riding with legs astride her mount. Taking seriously Bonheur's Parthenonian quotation, how should her self-portrait within the male-dominated arena of the horse market be understood? The author argues that, by classical analogy, Bonheur may be regarded as a gender-bending Amazon of a sort that was radically distinct from the scores of so-called "amazones" promenading about Paris. A comparative consideration of contemporary visualizations of the Amazonian rider trope suggests that Bonheur appropriates and, as it were, refashions this modish, gendered imagery to make a bold statement of women's equality with men.

Catalytic Antibodies May Contribute to Demyelination in Myalgic Encephalomyelitis/Chronic Fatigue Syndrome.

Here we report preliminary data demonstrating that some patients with myalgic encephalomyelitis/chronic fatiguesyndrome (ME/CFS) may have catalytic autoantibodies that cause the breakdown of myelin basic protein (MBP). We propose that these MBP-degradative antibodies are important to the pathophysiology of ME/CFS, particularly in the occurrence of white matter disease/demyelination. This is supported by magnetic resonance imagining studies that show these findings in patients with ME/CFS and could explain symptoms of nerve pain and muscle weakness. In this work, we performed a series of experiments on patient plasma samples where we isolated and characterized substrate-specific antibodies that digest MBP. We also tested glatiramer acetate (copaxone), an FDA approved immunomodulator to treat multiple sclerosis, and found that it inhibits ME/CFS antibody digestion of MBP. Furthermore, we found that aprotinin, which is a specific serine protease inhibitor, specifically prevents breakdown of MBP while the other classes of protease inhibitors had no effect. This coincides with the published literature describing catalytic antibodies as having serine protease-like activity. Postpandemic research has also provided several reports of demyelination in COVID-19. Because COVID-19 has been described as a trigger for ME/CFS, demyelination could play a bigger role in patient symptoms for those recently diagnosed with ME/CFS. Therefore, by studying proteolytic antibodies in ME/CFS, their target substrates, and inhibitors, a new mechanism of action could lead to better treatment and a possible cure for the disease.

Knowledge, attitudes, and practices in colorectal cancer screening in the Philippines

Background and Objective@#Colorectal cancer (CRC) has the third highest incidence in the Philippines. Currently, there is a paucity in literature that is focused on the knowledge, attitudes, and perceptions of Filipinos regarding CRC screening. This is the first study in the Philippines that describes this. @*Methods@#This is a cross-sectional study that validated a 52-item Filipino questionnaire on the knowledge on colorectal cancer, willingness to undergo CRC screening, and perceived benefits and barriers to fecal occult blood test (FOBT) and colonoscopy. The study enrolled household heads more than 20 years of age residing in both urban and rural communities in the Philippines. @*Results@#The UP-PGH CRC KAP (University of the Philippines – Philippine General Hospital Colorectal Cancer Knowledge, Attitudes, and Practices) and Rawl Questionnaire’s validity and internal consistency were established in a pilot study of 30 respondents. A total of 288 respondents were then enrolled to the main study group with a median age of 54.0. Knowledge scores for prognosis and utility of CRC screening were modest (6.3/12 and 8.4/20, respectively). Perceived benefit scores to FOBT and colonoscopy were high (9.9/12 and 13.9/16, respectively).Median scores to barriers to FOBT and colonoscopy were intermediate (22.5/36 and 35.8/60, respectively). Notably, a vast majority (86.1%) were willing to participate in CRC screening programs initiated by the government, and 46.9% agreed to undergo screening tests even as out-of-pocket expense. @*Conclusion@#The UP-PGH CRC KAP Questionnaire as well as the Filipino translation of the Rawl Questionnaire are reliable and valid tools in extensively assessing the knowledge of Filipinos on CRC and willingness to undergo screening, as well as the benefits of and barriers to FOBT and colonoscopy. Knowledge scores were modest suggesting that directed educational campaigns and awareness programs can aid in increasing awareness about CRC and its screening. Household income and highest educational attainment were significantly positively correlated with knowledge scores, and perceived benefits of and barriers to CRC screening. Scores were generally comparable between urban and rural communities.

Multivariate analysis of FcR-mediated NK cell functions identifies unique clustering among humans and rhesus macaques.

Rhesus macaques (RMs) are a common pre-clinical model used to test HIV vaccine efficacy and passive immunization strategies. Yet, it remains unclear to what extent the Fc-Fc receptor (FcR) interactions impacting antiviral activities of antibodies in RMs recapitulate those in humans. Here, we evaluated the FcR-related functionality of natural killer cells (NKs) from peripheral blood of uninfected humans and RMs to identify intra- and inter-species variation. NKs were screened for FcγRIIIa (human) and FcγRIII (RM) genotypes (FcγRIII(a)), receptor signaling, and antibody-dependent cellular cytotoxicity (ADCC), the latter mediated by a cocktail of monoclonal IgG1 antibodies with human or RM Fc. FcγRIII(a) genetic polymorphisms alone did not explain differences in NK effector functionality in either species cohort. Using the same parameters, hierarchical clustering separated each species into two clusters. Importantly, in principal components analyses, ADCC magnitude, NK contribution to ADCC, FcγRIII(a) cell-surface expression, and frequency of phosphorylated CD3ζ NK cells all contributed similarly to the first principal component within each species, demonstrating the importance of measuring multiple facets of NK cell function. Although ADCC potency was similar between species, we detected significant differences in frequencies of NK cells and pCD3ζ+ cells, level of cell-surface FcγRIII(a) expression, and NK-mediated ADCC (P<0.001), indicating that a combination of Fc-FcR parameters contribute to overall inter-species functional differences. These data strongly support the importance of multi-parameter analyses of Fc-FcR NK-mediated functions when evaluating efficacy of passive and active immunizations in pre- and clinical trials and identifying correlates of protection. The results also suggest that pre-screening animals for multiple FcR-mediated NK function would ensure even distribution of animals among treatment groups in future preclinical trials.

Case Report: Traumatic Distal Achilles Tendon Rupture in a Pediatric Patient.

Introduction: Acute Achilles tendon ruptures in the pediatric population are an uncommon occurrence and there is currently limited literature with regard to this injury. Further, there is limited data evaluating traumatic lacerations of the distal substance of the Achilles tendon near the insertion site and its management, as opposed to the more common cited location of rupture at the mid-substance of the tendon. This case is the first known article to specifically address primary repair with non-absorbable sutures utilizing a Krackow configuration at this level in this patient population. Case Report: A case of a 10-year-old male who sustained a traumatic distal Achilles tendon laceration near the insertion site who was successfully treated with primary repair with non-absorbable sutures utilizing a Krackow configuration is presented. This repair technique demonstrated return to full functional ankle plantar and dorsiflexion with return to pre-injury level of activity. Conclusion: This case report highlights that primary repair with non-absorbable sutures utilizing a Krackow configuration on distal traumatic Achilles tendon lacerations near the insertion site in pediatric patients is a safe and effective method of repair for this uncommon injury presentation.

Prehospital early warning scores for adults with suspected sepsis: retrospective diagnostic cohort study.

BACKGROUND: Ambulance services need to identify and prioritise patients with sepsis for early hospital assessment. We aimed to determine the accuracy of early warning scores alongside paramedic diagnostic impression to identify sepsis that required urgent treatment. METHODS: We undertook a retrospective diagnostic cohort study involving adult emergency medical cases transported to Sheffield Teaching Hospitals ED by Yorkshire Ambulance Service in 2019. We used routine ambulance service data to calculate 21 early warning scores and categorise paramedic diagnostic impressions as sepsis, infection, non-specific presentation or other presentation. We linked cases to hospital records and identified those meeting the sepsis-3 definition who received urgent hospital treatment for sepsis (reference standard). Analysis determined the accuracy of strategies that combined early warning scores at varying thresholds for positivity with paramedic diagnostic impression. RESULTS: We linked 12 870/24 955 (51.6%) cases and identified 348/12 870 (2.7%) with a positive reference standard. None of the strategies provided sensitivity greater than 0.80 with positive predictive value greater than 0.15. The area under the receiver operating characteristic curve for the National Early Warning Score, version 2 (NEWS2) applied to patients with a diagnostic impression of sepsis or infection was 0.756 (95% CI 0.729, 0.783). No other early warning score provided clearly superior accuracy to NEWS2. Paramedic impression of sepsis or infection had sensitivity of 0.572 (0.519, 0.623) and positive predictive value of 0.156 (0.137, 0.176). NEWS2 thresholds of >4, >6 and >8 applied to patients with a diagnostic impression of sepsis or infection, respectively, provided sensitivities and positive predictive values of 0.522 (0.469, 0.574) and 0.216 (0.189, 0.245), 0.447 (0.395, 0.499) and 0.274 (0.239, 0.313), and 0.314 (0.268, 0.365) and 0.333 (0.284, 0.386). CONCLUSION: No strategy is ideal but using NEWS2 alongside paramedic diagnostic impression of infection or sepsis could identify one-third to half of sepsis cases without prioritising unmanageable numbers. No other score provided clearly superior accuracy to NEWS2. TRIAL REGISTRATION NUMBER: researchregistry5268, https://www.researchregistry.com/browse-the-registry%23home/registrationdetails/5de7bbd97ca5b50015041c33/.

Microparticle electrical conductivity measurement using optoelectronic tweezers.

When it comes to simulate or calculate an optoelectronic tweezer (OET) response for a microparticle suspended in a given medium, a precise electrical conductivity (later referred to as conductivity) value for the microparticle is critical. However, there are not well-established measurements or well-referenced values for microparticle conductivities in the OET realm. Thus, we report a method based on measuring the escape velocity of a microparticle with a standard OET system to calculate its conductivity. A widely used 6 µm polystyrene bead (PSB) is used for the study. The conductivity values are found to be invariant around 2×10-3 S/m across multiple different aqueous media, which helps clarify the ambiguity in the usage of PSB conductivity. Our convenient approach could principally be applied for the measurement of multiple unknown OET-relevant material properties of microparticle-medium systems with various OET responses, which can be beneficial to carry out more accurate characterization in relevant fields.

Mpox Pain Management with Topical Agents: A Case Series.

With the recent spread in monkeypox cases, continuous efforts are made to manage the disease efficiently. Pain at the site of monkeypox lesions and in areas of skin breakdown can be severe. The origin of pain is likely neuropathic. The Centers for Disease Control and Prevention (CDC) has issued general guidelines to control pain with non-steroidal anti-inflammatory drugs (NSAIDs), acetaminophen, gabapentin, and topical agents such as corticosteroids and lidocaine. Guidelines circulated by the World Health Organization (WHO) suggest acetaminophen, tramadol, opioids, and/or topical lidocaine for symptomatic pain management. No first line agent with proven beneficial effect has been recommended in these patients. We present two patients' painful experiences with Monkeypox and suggest lessons learned.

Neofunctionalization of S-adenosylmethionine decarboxylase into pyruvoyl-dependent L-ornithine and L-arginine decarboxylases is widespread in bacteria and archaea.

S-adenosylmethionine decarboxylase (AdoMetDC/SpeD) is a key polyamine biosynthetic enzyme required for conversion of putrescine to spermidine. Autocatalytic self-processing of the AdoMetDC/SpeD proenzyme generates a pyruvoyl cofactor from an internal serine. Recently, we discovered that diverse bacteriophages encode AdoMetDC/SpeD homologs that lack AdoMetDC activity and instead decarboxylate L-ornithine or L-arginine. We reasoned that neofunctionalized AdoMetDC/SpeD homologs were unlikely to have emerged in bacteriophages and were probably acquired from ancestral bacterial hosts. To test this hypothesis, we sought to identify candidate AdoMetDC/SpeD homologs encoding L-ornithine and L-arginine decarboxylases in bacteria and archaea. We searched for the anomalous presence of AdoMetDC/SpeD homologs in the absence of its obligatory partner enzyme spermidine synthase, or the presence of two AdoMetDC/SpeD homologs encoded in the same genome. Biochemical characterization of candidate neofunctionalized genes confirmed lack of AdoMetDC activity, and functional presence of L-ornithine or L-arginine decarboxylase activity in proteins from phyla Actinomycetota, Armatimonadota, Planctomycetota, Melainabacteria, Perigrinibacteria, Atribacteria, Chloroflexota, Sumerlaeota, Omnitrophota, Lentisphaerota, and Euryarchaeota, the bacterial candidate phyla radiation and DPANN archaea, and the δ-Proteobacteria class. Phylogenetic analysis indicated that L-arginine decarboxylases emerged at least three times from AdoMetDC/SpeD, whereas L-ornithine decarboxylases arose only once, potentially from the AdoMetDC/SpeD-derived L-arginine decarboxylases, revealing unsuspected polyamine metabolic plasticity. Horizontal transfer of the neofunctionalized genes appears to be the more prevalent mode of dissemination. We identified fusion proteins of bona fide AdoMetDC/SpeD with homologous L-ornithine decarboxylases that possess two, unprecedented internal protein-derived pyruvoyl cofactors. These fusion proteins suggest a plausible model for the evolution of the eukaryotic AdoMetDC.

Exploring Stroke Risk Factors and Outcomes in Sexual and Gender Minority People.

Background and Objectives: Cerebrovascular disease in sexual and gender minority (SGM) people remains poorly understood. Our primary objective was to describe the epidemiology and outcomes in a sample of SGM people with stroke. As a secondary objective, we compared this group with non-SGM people with stroke to assess for significant differences in risk factors or outcomes. Methods: This was a retrospective chart review study of SGM people admitted to an urban stroke center with primary diagnosis of stroke (ischemic or hemorrhagic). We evaluated stroke epidemiology and outcomes, summarizing with descriptive statistics. We then matched 1 SGM person to 3 non-SGM people by year of birth and year of diagnosis to compare demographics, risk factors, inpatient stroke metrics, and outcomes. Results: A total of 26 SGM people were included in the analysis: 20 (77%) had ischemic strokes, 5 (19%) intracerebral hemorrhages, and 1 (4%) subarachnoid hemorrhage. Compared with non-SGM people (n = 78), stroke subtypes showed a similar distribution (64 (82%) ischemic strokes, 12 (15%) intracerebral hemorrhages, 1 (1%) subarachnoid hemorrhage, and 1 nontraumatic subdural hematoma, p > 0.05) but suspected ischemic stroke mechanisms had a different distribution (χ2 = 17.56, p = 0.01). Traditional stroke risk factors were similar between the 2 groups. The SGM group seemed to have higher rates of nontraditional stroke factors, including HIV (31% vs 0%, p < 0.01), syphilis (19% vs 0%, p < 0.01), and hepatitis C (15% vs 5%, p < 0.01) but were more likely to be tested for these risk factors (χ2 = 15.80, p < 0.01; χ2 = 11.65, p < 0.01; χ2 = 7.83, p < 0.01, respectively). SGM people were more likely to have recurrent strokes (χ2 = 4.39, p < 0.04) despite similar follow-up rates. Discussion: SGM people may have different risk factors, different mechanisms of stroke, and higher risk of recurrent stroke compared with non-SGM people. Standardized collection of sexual orientation and gender identity would enable larger studies to further understand disparities, leading to secondary prevention strategies.

Long-term outcome of cervical artery dissection.

OBJECTIVE: The aim of the study is to evaluate the natural history of extracranial cervical artery dissection (CAD) including comorbidities, symptoms at presentation, recurrence of symptoms, and long-term outcome following different treatment approaches. METHODS: A retrospective review of patients treated for acute CAD was performed over a 5-year period from January 2017 to April 2022. RESULTS: Thirty-nine patients were included in the study, 25 (64.1%) with acute internal carotid artery dissection and 14 (35.9%) with acute vertebral artery dissection. Thirty-four patients (87.1%) had spontaneous CAD, and five patients (12.8%) had traumatic CAD. The mean age of the cohort was 54.2 years. The mean time from symptom onset to presentation was 4.34 days. The most common symptoms in internal carotid artery dissection were unilateral weakness (44%), headache (44%), slurred speech (36%), facial droop (28%), unilateral paraesthesia (24%), neck pain (12%), visual disturbance (8%), and Horner's syndrome (8%). The most common symptoms in vertebral artery dissection were headache (35.7%), neck pain (35.7%), vertigo (28.57%), ataxia (14.28%), and slurred speech (14.28%). The imaging modalities used for diagnosis included computed tomography angiography (48.7%), magnetic resonance angiography (41%), and duplex ultrasound (10.2%). In patients with carotid artery dissection, 57% had severe stenosis, 24% had moderate stenosis, and 20% had mild stenosis. All patients treated were managed conservatively with either anticoagulation or antiplatelets. Long-term clinical follow-up was available for 33 patients (84.6%). Thirty patients (90.9%) reported complete resolution of symptoms, and three patients (9%) reported persistent symptoms. Anatomic follow-up with imaging was available for 17 patients (43.58%). Thirteen patients (76.47%) had complete resolution of dissection, two patients (11.76%) had partial resolution of dissection, and two patients (11.76%) had persistent dissection. There was one death unrelated to CAD in a multi-trauma patient. There were four early recurrent symptoms in the first 3 to 8 weeks post discharge. The mean follow-up time was 308.27 days. CONCLUSIONS: The majority of CADs can be managed conservatively with good clinical and anatomical outcome and low rates of recurrence.

Modified T2 Statistics for Improved Detection of Aided Cortical Auditory Evoked Potentials in Hearing-Impaired Infants.

The cortical auditory evoked potential (CAEP) is a change in neural activity in response to sound, and is of interest for audiological assessment of infants, especially those who use hearing aids. Within this population, CAEP waveforms are known to vary substantially across individuals, which makes detecting the CAEP through visual inspection a challenging task. It also means that some of the best automated CAEP detection methods used in adults are probably not suitable for this population. This study therefore evaluates and optimizes the performance of new and existing methods for aided (i.e., the stimuli are presented through subjects' hearing aid(s)) CAEP detection in infants with hearing loss. Methods include the conventional Hotellings T2 test, various modified q-sample statistics, and two novel variants of T2 statistics, which were designed to exploit the correlation structure underlying the data. Various additional methods from the literature were also evaluated, including the previously best-performing methods for adult CAEP detection. Data for the assessment consisted of aided CAEPs recorded from 59 infant hearing aid users with mild to profound bilateral hearing loss, and simulated signals. The highest test sensitivities were observed for the modified T2 statistics, followed by the modified q-sample statistics, and lastly by the conventional Hotelling's T2 test, which showed low detection rates for ensemble sizes <80 epochs. The high test sensitivities at small ensemble sizes observed for the modified T2 and q-sample statistics are especially relevant for infant testing, as the time available for data collection tends to be limited in this population.

Functional polyamine metabolic enzymes and pathways encoded by the virosphere.

Viruses produce more viruses by manipulating the metabolic and replication systems of their host cells. Many have acquired metabolic genes from ancestral hosts and use the encoded enzymes to subvert host metabolism. The polyamine spermidine is required for bacteriophage and eukaryotic virus replication, and herein, we have identified and functionally characterized diverse phage- and virus-encoded polyamine metabolic enzymes and pathways. These include pyridoxal 5'-phosphate (PLP)-dependent ornithine decarboxylase (ODC), pyruvoyl-dependent ODC and arginine decarboxylase (ADC), arginase, S-adenosylmethionine decarboxylase (AdoMetDC/speD), spermidine synthase, homospermidine synthase, spermidine N-acetyltransferase, and N-acetylspermidine amidohydrolase. We identified homologs of the spermidine-modified translation factor eIF5a encoded by giant viruses of the Imitervirales. Although AdoMetDC/speD is prevalent among marine phages, some homologs have lost AdoMetDC activity and have evolved into pyruvoyl-dependent ADC or ODC. The pelagiphages that encode the pyruvoyl-dependent ADCs infect the abundant ocean bacterium Candidatus Pelagibacter ubique, which we have found encodes a PLP-dependent ODC homolog that has evolved into an ADC, indicating that infected cells would contain both PLP- and pyruvoyl-dependent ADCs. Complete or partial spermidine or homospermidine biosynthetic pathways are found encoded in the giant viruses of the Algavirales and Imitervirales, and in addition, some viruses of the Imitervirales can release spermidine from the inactive N-acetylspermidine. In contrast, diverse phages encode spermidine N-acetyltransferase that can sequester spermidine into its inactive N-acetyl form. Together, the virome-encoded enzymes and pathways for biosynthesis and release or biochemical sequestration of spermidine or its structural analog homospermidine consolidate and expand evidence supporting an important and global role of spermidine in virus biology.