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It is difficult to simultaneously increase strength and toughness of austenitic stainless steels through traditional heat treatment and alloying. However, adding nano-sized ceramic particles is a potential method to manipulate the microstructure and mechanical properties of austenitic stainless steels. In this study, TiC–TiB2 nanoparticles were introduced into austenitic stainless steel using an aluminum master alloy. Adding 0.02 wt% nanoparticles effectively refined the substrates including the dendritic ferrite and austenite and reduced the grain size from 23.7 μm to 18.4 μm. The edge-to-edge model indicated that the nanoparticles act as heterogeneous cores to facilitate the nucleation rates. For mechanical properties, the addition of nanoparticles simultaneously enhanced the hardness, yield strength, tensile strength and impact toughness of the austenitic stainless steels without sacrificing the ductility. The yield strength and tensile strength of as-cast austenitic stainless steels were enhanced by 4.8% and 2.1%, 7.2% and 3.9%, 10.0% and 5.7% at the temperatures of 20 °C, 200 °C, and 400 °C, respectively. The impact toughness was improved by 4.2%, 5.9%, and 4.3% at the temperatures of −20 °C, 0 °C, and 20 °C. Also, the presence of nanoparticles increased the yield strength and tensile strength of forged austenitic stainless steels by 16.6% and 5.9%, 17.7% and 7.7%, 18.5% and 12.2% at the temperatures of 20 °C, 200 °C, and 400 °C, respectively. The impact toughness was increased by 11.6%, 8.6% and 11.3% at the temperatures of −20 °C, 0 °C, and 20 °C. The nanoparticles refined the grains, inhibited dislocation movement, scattered large cracks and distributed the external loads more uniformly to strengthen and toughen the austenitic stainless steels. In wear tests, the nanoparticle reinforced austenitic stainless steels showed higher wear resistance against adhesive wear and abrasive wear by impeding the nucleation and growth of cracks, retarding the dislocation migration and plastic deformation and bearing the external loads. The wear volumes of as-cast and forged austenitic stainless steels were reduce by 15.7% and 21.5%. This innovative approach provided a promising strategy to produce high-performance austenitic stainless steels and serve as a reference in the development of other nanoparticle reinforced ferrous materials.

Abstract Undifferentiated pleomorphic sarcoma (UPS) and related tumors are the most common type of soft tissue sarcoma. However, this spectrum of tumors has different etiologies with varying rates of metastasis and survival. Two dermal-based neoplasms in this class of pleomorphic sarcomas, atypical fibroxanthoma (AFX) and pleomorphic dermal sarcoma (PDS), are challenging to differentiate at initial biopsy but vary significantly in prognosis. We performed single-cell transcriptomics on five AFX and PDS biopsy specimens as well as both single-cell and spatial transcriptomics on one PDS excision specimen to better characterize these tumors. The top differential genes between AFX and PDS were predictive of overall survival in 17 other cancers included in the Human Protein Atlas. Of these genes, COL6A3 and BGN predicted overall survival and metastasis-free survival in independent cohorts of 46 and 38 UPS tumors, respectively. COL6A3 was most predictive of overall survival in UPS patients and outperformed an established sarcoma prognostic gene panel at predicting metastasis in UPS.

In this work, the resistance spot welding (RSW) process was performed to join aluminum alloy and low-alloy carbon steel plates. The macro characteristics including nugget diameters and indentation rates, microstructure and tensile-shear strength of RSW joints were investigated. The results showed that the nugget of the RSW joints comprises a ‘bowl’ shape nugget on the aluminum side and an elliptical shape nugget on the steel side. Also, the intermetallic compound (IMC) layers containing Fe4Al13 and Fe2Al5 were formed around the aluminum/steel interface with the steel side having a tongue-shape and the aluminum side having a needle-shape. According to metallurgical evaluation and temperature distribution of RSW joints analyzed by the finite element method, the nugget on the aluminum side contained the dendritic grains and equiaxed dendritic grains. The nugget on the steel side consisted of a large amount of bainite and a small amount of coarse lath martensite. The nugget diameters and the indentations rates of RSW joints increased when increasing either welding current or welding time, and decreased when increasing the electrode pressure. The maximum values of nugget diameter and indentation rate of RSW joints were 9.076 mm and 4.144% when using the welding current of 16 kA, welding time of 450 ms and electrode force of 3 kN. In tensile-shear tests, the RSW joints showed a shear-off fracture mode. When increasing the welding current, welding time or electrode force, the tensile-shear strength of RSW joints increased first, and then reached a maximum, and finally decreased. The welding current of 16 kA, the welding time of 300 ms, and the electrode pressure of 3 kN were considered as the optimal welding parameters in the present study which resulted in the maximum tensile-shear strength of 2.24 kN. In addition, the IMC layers of the RSW joints exhibited a uniform and continuous appearance with a thickness of approximately 1.9 μm, and the IMC layer in the central area was thicker than that in the edge area.

Abstract Background Herpes zoster (HZ), commonly known as “shingles,” may contribute to cognitive decline through mechanisms such as neuroinflammation or direct neuronal injury. However, evidence on the longitudinal association between HZ and cognitive decline is conflicting and whether the risk differs by APOE ε4-carrier status has not been studied; prospective cohort studies on the association between HZ vaccination and cognitive decline are also lacking. Methods We included 149,327 participants from three large cohorts—the Nurses’ Health Study (NHS), NHSII, and Health Professionals Follow-Up Study (HPFS)—to prospectively examine the association between HZ and subsequent subjective cognitive decline (SCD). Poisson regression was used to estimate the multivariable-adjusted relative risk (MVRR) of a 3-unit increment in SCD score according to years since HZ compared with participants with no history of HZ. Results Compared with individuals with no history of HZ, the MVRR (95% CI) of a 3-unit increment in SCD score was significantly and independently higher among individuals with a history of HZ, but the duration of time since HZ when the elevated risk of SCD was statistically significant differed among the cohorts. In NHS, HZ was associated with higher long-term risk of SCD; compared with individuals with no history of HZ, the MVRR (95% CI) of a 3-unit increment in SCD score was 1.14 (1.01, 1.32) for ≥ 13 years since HZ. In NHS II, HZ was associated with higher risk of SCD in both the short-term [MVRR 1.34 (1.18, 1.53) for 1–4 years] and long-term [MVRR 1.20 (1.08, 1.34) for ≥ 13 years since HZ]. In HPFS, an elevated risk of SCD was suggested across all time points. Among the subset of participants with information on APOE ε4, there was a suggestion that the association differed by APOE ε4 carrier status, but the results were not consistent between women and men. Among the subset of women with information on HZ vaccination, there was a suggestion that the long-term risk of SCD may be greater among women who were not vaccinated against HZ. Conclusions Data from three large independent cohorts of women and men showed that HZ was associated with higher long-term risk of SCD, and the risk may differ by APOE ε4-carrier status.

Abstract Foliar fungal diseases are a major limitation in organic naked barley (Hordeum vulgare L.) production. The lack of conventional fungicides in organic systems increases reliance on genetic resistance. We evaluated the severity of barley stripe rust (Puccinia striiformis f. sp. hordei Westend), leaf rust (Puccina hordei sp. hordei), spot blotch (Cochliobolus sativus, anamorph Bipolaris sorokiniana (S. Ito & Kurib.) Drechsler ex Dastur), and scald (Rhynchosporium commune Zaffarano, McDonald and Linde sp. nov) on a naked barley diversity panel of 350 genotypes grown in 13 environments to identify quantitative trait loci associated with disease resistance. Genome‐wide association analyses across and within environments found 10 marker trait associations for barley stripe rust, four marker trait associations for leaf rust, one marker trait association for scald, and five marker trait associations for spot blotch. Structure analysis identified six Ward groups based on genotypic diversity. Resistance to susceptible allele ratios were high for stripe rust and spot blotch, moderate for leaf rust, and low for scald. Combined phenotypic analysis values for each disease overlayed by a principal component analysis found distinct resistance and susceptibility patterns for barley stripe rust and scald. Most significant marker trait associations were previously identified in the literature, providing confirmation and potential new sources of disease resistance for genetic improvement of naked barley germplasm.

Background: Clinical trial participation can improve overall survival and mitigate healthcare disparities for gynecologic cancer patients in low-volume community centers. This study aimed to assess the effectiveness of a centrally regulated but administratively decentralized electronic screening log system to identify eligible patients across a large catchment area for a National Cancer Institute (NCI)-designated cancer center's open clinical trials. Methods: Electronic screening log data collected between 2014 and 2021 from ten community partner sites in a single NCI-designated cancer center's catchment area were reviewed retrospectively. Clinical factors assessed included cancer site, primary versus recurrent disease status, and histology. Identification efficiency (the ratio of patients screened identified with an available trial) was calculated. Identification inefficiencies (failures to identify patients with a potentially relevant trial) were assessed, and etiologies were characterized. Results: Across ten community partner sites, 492 gynecologic cancer patients were screened for seven open clinical trials during the study period. This included 170 (34.5 %) ovarian cancer patients, 156 (31.7 %) endometrial cancer patients, and 119 (24.2 %) cervical cancer patients. Over 40 % had advanced stage disease, and 10.6 % had recurrent disease. Only three patients were identified as having a relevant open trial; none ultimately enrolled due to not meeting trial eligibility criteria. An additional 2–52 patients were retrospectively found to have a relevant trial available despite not being identified as such within the electronic screening log system. Up to 14.4 % of patients had one or more missing minimum data elements that hindered full evaluation of clinical trial availability. Re-screening patients when new trials open may identify 12-15 additional patients per recurrent disease trial. Conclusions: An electronic screening log system can increase awareness of gynecologic oncology clinical trials at a NCI-designated cancer center's community partner sites. However, it is inadequate as a single intervention to increase clinical trial enrollment. Providing adequate support staff, documenting clinical factors consistently, re-screening patients at relevant intervals, and coordinating with central study personnel may increase its utility.

ABSTRACT Background Cachexia accounts for about 20% of all cancer‐related deaths and indicates poor prognosis. The impact of Fusobacterium nucleatum (Fn), a microbial risk factor for colorectal cancer (CRC), on the development of cachexia in CRC has not been established. Methods We evaluated the association between Fn abundance in pre‐surgical stool samples and onset of cachexia at 6 months post‐surgery in n = 87 patients with stages I–III CRC in the ColoCare Study. Results High fecal Fn abundance compared to negative/low fecal Fn abundance was associated with 4‐fold increased risk of cachexia onset at 6 months post‐surgery (OR = 4.82, 95% CI = 1.15, 20.10, p = 0.03). Conclusion Our findings suggest that high fecal Fn abundance was associated with an increased risk of cachexia at 6 months post‐surgery in CRC patients. This is the first study to link Fn abundance with cachexia in CRC patients, offering novel insights into biological mechanisms and potential management of cancer cachexia. Due to the small sample size, our results should be interpreted with caution. Future studies with larger sample sizes are needed to validate these findings.

Dengue virus (DENV) is the causative agent of dengue, a mosquito-borne disease that represents a significant and growing public health burden around the world. A unique pathophysiological feature of dengue is immune-mediated enhancement, wherein preexisting immunity elicited by a primary infection can enhance the severity of a subsequent infection by a heterologous DENV serotype. A leading mechanistic explanation for this phenomenon is antibody dependent enhancement (ADE), where sub-neutralizing concentrations of DENV-specific IgG antibodies facilitate entry of DENV into FcγR expressing cells such as monocytes, macrophages, and dendritic cells. Accordingly, this model posits that phagocytic mononuclear cells are the primary reservoir of DENV. However, analysis of samples from individuals experiencing acute DENV infection reveals that B cells are the largest reservoir of infected circulating cells, representing a disconnect in our understanding of immune-mediated DENV tropism. In this study, we demonstrate that the expression of a DENV-specific B cell receptor (BCR) renders cells highly susceptible to DENV infection, with the infection-enhancing activity of the membrane-restricted BCR correlating with the ADE potential of the IgG version of the antibody. In addition, we observed that the frequency of DENV-infectible B cells increases in previously flavivirus-naïve volunteers after a primary DENV infection. These findings suggest that BCR-dependent infection of B cells is a novel mechanism immune-mediated enhancement of DENV-infection.

Large scale cancer genomic studies in patients have unveiled millions of non-coding variants. While a handful have been shown to drive cancer development, the vast majority have unknown function. This review describes the challenges of functionally annotating non-coding cancer variants and understanding how they contribute to cancer. We summarize recently developed high-throughput technologies to address these challenges. Finally, we outline future prospects for non-coding cancer genetics to help catalyze personalized cancer therapy.

Abstract Succinate dehydrogenase (SDH) is a key mitochondrial enzyme involved in the tricarboxylic acid cycle, where it facilitates the oxidation of succinate to fumarate, and is coupled to the reduction of ubiquinone in the electron transport chain as Complex II. Previously, we developed a confocal‐based quantitative histochemical technique to determine the maximum velocity of the SDH reaction (SDHmax) in single cells and observed that SDHmax corresponds with mitochondrial volume density. In addition, mitochondrial volume and motility varied within different compartments of human airway smooth muscle (hASM) cells. Therefore, we hypothesize that the SDH activity varies relative to the intracellular mitochondrial volume within hASM cells. Using 3D confocal imaging of labeled mitochondria and a concentric shell method for analysis, we quantified mitochondrial volume density, mitochondrial complexity index, and SDHmax relative to the distance from the nuclear membrane. The mitochondria within individual hASM cells were more filamentous in the immediate perinuclear region and were more fragmented in the distal parts of the cell. Within each shell, SDHmax also corresponded to mitochondrial volume density, where both peaked in the perinuclear region and decreased in more distal parts of the cell. Additionally, when normalized to mitochondrial volume, SDHmax was lower in the perinuclear region when compared to the distal parts of the cell. In summary, our results demonstrate that SDHmax measures differences in SDH activity within different cellular compartments. Importantly, our data indicate that mitochondria within individual cells are morphologically heterogeneous, and their distribution varies substantially within different cellular compartments, with distinct functional properties.

The current study innovatively proposed to prepare TiC nanoparticles reinforced high-Cr hot work die steels through an aluminum master alloy. The influences of TiC nanoparticles on the microstructure and wear resistance of high-Cr hot work die steels were thoroughly investigated. It was found that the addition of nanoparticles refined the pro-eutectoid ferrite in the as-cast condition, and the granular pearlite in the isothermal spheroidized condition, respectively. After receiving the quenching and tempering heat treatment, the size of lath martensite, retained austenite and nanosized precipitates were reduced. The TiC nanoparticles were used as heterogeneous cores to improve the nucleation rates of austenite and ferrite during phase transformation. The finer grains with more grain boundaries and nanoparticles refined the size of tempered microstructure of high-Cr hot work die steels. In ball-on-plate reciprocating sliding wear tests, it was found that the presence of TiC nanoparticles enhanced the wear resistance of high-Cr hot work die steels under various test conditions. The nanoparticles increased the strength of the substrate and inhibited the formation of cracks. More grain boundaries and nanoparticles hindered the migration of dislocations, which resulted in less plastic deformation. Also, the nanoparticles could partially withstand the external load during the relative motion. The results obtained in this research provide applicable values for optimizing the strength-ductility and tribological performance of hot work die steels.

Abstract In today's world, remote-controlled robots are widely used across various industries due to their ability to enhance working efficiency in various applications. Learning about robot operation and human–computer interaction has emerged as a popular topic in recent times. Indeed, learning robotics can be challenging for many students as it requires knowledge of programming, control systems, electronics, etc. Collaborative learning in a physical robotics setting is common in higher education and has received significant attention for its potential to enhance individual learning outcomes. However, the effectiveness of learning robotics in a remote setting is still a matter of debate. In this study, we establish a remote laboratory environment to teach undergraduate students in the engineering discipline. Students are required to utilize a robotic arm to grasp designated objects collaboratively among students through synchronous interactions online. To compare students’ performance under different pedagogical teaching approaches, students are divided into two groups. They each perform the task individually and collaboratively, albeit in a different order. Our study adopts a quantitative method to measure students' learning outcomes based on the assessment of performing the laboratory tasks and completion time. The results indicate a noteworthy improvement in the individual performance of the group of students who engage in collaborative work prior to the individual tasks. These findings have implications for other remote laboratory setups and highlight the effectiveness of collaborative learning in higher education.

The UK Biobank includes genotype information for about 500,000 patients for over 7000 phenotypes. However, owing to multiple testing correction for approximately 200 billion tests, many clinically and statistically significant associations remain unappreciated. We perform a focused analysis of the UK Biobank for 13 dermatologic conditions, including malignant melanoma, melanoma in situ, squamous cell carcinoma, basal cell carcinoma, actinic keratosis, seborrheic keratosis, psoriasis, lichen planus, systemic lupus erythematosus, hyperhidrosis, pilonidal cyst, sebaceous cyst, and lipoma. We identify 447 sentinel variants, which are enriched for protein-coding variants and an elevated combined annotation-dependent depletion (CADD) score compared with background variants. Through gene ontology enrichment analysis, we identify known pathways involved in melanoma, actinic keratoses, and squamous cell carcinoma and uncover additional pathways. We also uncover 5 protein-coding variants, which, to our knowledge, have not been previously reported, including LRP3 for lipomas, PLCD1 for sebaceous cysts, EIF3CL for lichen planus, TTK for pilonidal cysts, and MAPK15 for systemic lupus erythematosus.

Autophagy is a ubiquitous process through which damaged cytoplasmic structures are recycled and degraded within cells. Aging can affect autophagy regulation in different steps leading to the accumulation of damaged organelles and proteins, which can contribute to cell dysfunction and death. Motor neuron (MN) loss and sarcopenia are prominent features of neuromuscular aging. Previous studies on phrenic MNs showed increased levels of the autophagy proteins LC3 and p62 in 24 month compared to 6 month old mice, consistent with the onset of diaphragm muscle sarcopenia. In the present study, we hypothesized that aging leads to increased expression of the autophagy markers LC3 and p62 in single lumbar MNs. Expression of LC3 and p62 in lumbar MNs (spinal levels L1-L6) was assessed using immunofluorescence and confocal imaging of male and female mice at 6, 18 and 24 months of age, reflecting 100 %, 90 % and 75 % survival, respectively. A mixed linear model with animal as a random effect was used to compare relative LC3 and p62 expression in choline acetyl transferase-positive MNs across age groups. Expression of LC3 and p62 decreased in the white matter of the lumbar spinal cord with aging, with ~29 % decrease in LC3 and ~ 7 % decrease in p62 expression at 24 months of age compared to 6 months of age. There was no change in LC3 or p62 expression in the gray matter with age. LC3 expression in MNs relative to white matter increased significantly with age, with 150 % increase at 24 months of age compared to 6 months of age. Similarly, p62 expression in MNs relative to white matter increased significantly with age, with ~14 % increase at 24 months of age compared to 6 months of age. No effect of sex or MN pool was observed in LC3 and p62 expression in MNs. Overall, these data suggest autophagy impairment during elongation (increased LC3) and degradation (increased p62) phases with aging in lumbar MNs.

Abstract Aging is associated with cardiac contractile abnormalities, but the etiology of these contractile deficits is unclear. We hypothesized that cardiac contractile and regulatory protein expression is altered during aging. To investigate this possibility, left ventricular (LV) lysates were prepared from young (6 months) and old (24 months) Fischer344 rats. There are no age‐related changes in SERCA2 expression or phospholamban phosphorylation. Additionally, neither titin isoform expression nor phosphorylation differed. However, there is a significant increase in β‐isoform of the myosin heavy chain (MyHC) expression and phosphorylation of TnI and MyBP‐C during aging. In permeabilized strips of papillary muscle, force and Ca2+ sensitivity are reduced during aging, consistent with the increase in β‐MyHC expression and TnI phosphorylation. However, the increase in MyBP‐C phosphorylation during aging may represent a mechanism to compensate for age‐related contractile deficits. In isolated cardiomyocytes loaded with Fura‐2, the peak of the Ca2+ transient is reduced, but the kinetics of the Ca2+ transient are not altered. Furthermore, the extent of shortening and the rates of both sarcomere shortening and re‐lengthening are reduced. These results demonstrate that aging is associated with changes in contractile and regulatory protein expression and phosphorylation, which affect the mechanical properties of cardiac muscle.

Abstract Immune checkpoints (CTLA4 & PD-1) are inhibitory pathways that block aberrant immune activity and maintain self-tolerance. Tumors co-opt these checkpoints to avoid immune destruction. Immune checkpoint inhibitors (ICIs) activate immune cells and restore their tumoricidal potential, making them highly efficacious cancer therapies. However, immunotolerant organs such as the liver depend on these tolerogenic mechanisms, and their disruption with ICI use can trigger the unintended side effect of hepatotoxicity termed immune-mediated liver injury from ICIs (ILICI). Learning how to uncouple ILICI from ICI anti-tumor activity is of paramount clinical importance. We developed a murine model to recapitulate human ILICI using CTLA4+/- mice treated with either combined anti-CTLA4 + anti-PDL1 or IgG1 + IgG2. We tested two forms of antisense oligonucleotides to knockdown caspase-3 in a total liver (parenchymal and non-parenchymal cells) or in a hepatocyte-specific manner. We also employed imaging mass cytometry (IMC), a powerful multiplex modality for immunophenotyping and cell interaction analysis in our model. ICI-treated mice had significant evidence of liver injury. We detected cleaved caspase-3 (cC3), indicating apoptosis was occurring, as well as Nod-like receptor protein 3 (NLRP3) inflammasome activation, but no necroptosis. Total liver knockdown of caspase-3 worsened liver injury, and induced further inflammasome activation, and Gasdermin-D-mediated pyroptosis. Hepatocyte-specific knockdown of caspase-3 reduced liver injury and NLRP3 inflammasome activation. IMC-generated single-cell data for 77,692 cells was used to identify 22 unique phenotypic clusters. Spatial analysis revealed that cC3+ hepatocytes had significantly closer interactions with macrophages, Kupffer cells, and NLRP3hi myeloid cells than other cell types. We also observed zones of three-way interaction between cC3+ hepatocytes, CD8 + T-cells, and macrophages. Our work is the first to identify hepatocyte apoptosis and NLRP3 inflammasome activation as drivers of ILICI. Furthermore, we report that the interplay between adaptive and innate immune cells is critical to hepatocyte apoptosis and ILICI.

Background Accurate quantification of sodium intake based on self‐reported dietary assessments has been a persistent challenge. We aimed to apply machine‐learning (ML) algorithms to predict 24‐hour urinary sodium excretion from self‐reported questionnaire information. Methods and Results We analyzed 3454 participants from the NHS (Nurses' Health Study), NHS‐II (Nurses' Health Study II), and HPFS (Health Professionals Follow‐Up Study), with repeated measures of 24‐hour urinary sodium excretion over 1 year. We used an ensemble approach to predict averaged 24‐hour urinary sodium excretion using 36 characteristics. The TOHP‐I (Trial of Hypertension Prevention I) was used for the external validation. The final ML algorithms were applied to 167 920 nonhypertensive adults with 30‐year follow‐up to estimate confounder‐adjusted hazard ratio (HR) of incident hypertension for predicted sodium. Averaged 24‐hour urinary sodium excretion was better predicted and calibrated with ML compared with the food frequency questionnaire (Spearman correlation coefficient, 0.51 [95% CI, 0.49–0.54] with ML; 0.19 [95% CI, 0.16–0.23] with the food frequency questionnaire; 0.46 [95% CI, 0.42–0.50] in the TOHP‐I). However, the prediction heavily depended on body size, and the prediction of energy‐adjusted 24‐hour sodium excretion was modestly better using ML. ML‐predicted sodium was modestly more strongly associated than food frequency questionnaire‐based sodium in the NHS‐II (HR comparing Q5 versus Q1, 1.48 [95% CI, 1.40–1.56] with ML; 1.04 [95% CI, 0.99–1.08] with the food frequency questionnaire), but no material differences were observed in the NHS or HPFS. Conclusions The present ML algorithm improved prediction of participants' absolute 24‐hour urinary sodium excretion. The present algorithms may be a generalizable approach for predicting absolute sodium intake but do not substantially reduce the bias stemming from measurement error in disease associations.

Introduction: Vasodilatation in response to NO is a fundamental response of the vasculature, and during aging, the vasculature is characterized by an increase in stiffness and decrease in sensitivity to NO mediated vasodilatation. Vascular tone is regulated by the activation of smooth muscle and nonmuscle (NM) myosin, which are regulated by the activities of myosin light chain kinase (MLCK) and MLC phosphatase. MLC phosphatase is a trimeric enzyme with a catalytic subunit, myosin targeting subunit (MYPT1) and 20 kDa subunit of unknown function. Alternative mRNA splicing produces LZ+/LZ- MYPT1 isoforms and the relative expression of LZ+/LZ- MYPT1 determines the sensitivity to NO mediated vasodilatation. This study tested the hypothesis that aging is associated with changes in LZ+ MYPT1 and NM myosin expression, which alter vascular reactivity.Methods: We determined MYPT1 and NM myosin expression, force and the sensitivity of both endothelial dependent and endothelial independent relaxation in tertiary mesenteric arteries of young (6mo) and elderly (24mo) Fischer344 rats.Results: The data demonstrate that aging is associated with a decrease in both the expression of NM myosin and force, but LZ+ MYPT expression and the sensitivity to both endothelial dependent and independent vasodilatation did not change. Further, smooth muscle cell hypertrophy increases the thickness of the medial layer of smooth muscle with aging.Discussion: The reduction of NM myosin may represent an aging associated compensatory mechanism to normalize the stiffness of resistance vessels in response to the increase in media thickness observed during aging.

Six new species of Ampeliscidae are described: Ampelisca capella sp. nov., Ampelisca mingela sp. nov., Byblis pialba sp. nov. and Byblis wadara sp. nov. from the Great Barrier Reef; Ampelisca katoomba sp. nov. from the New South Wales shelf; and Byblis liena sp. nov. from the Queensland slope. Byblisoides esferis J. L. Barnard, 1961 is recorded from southeastern Australia, the first record of the genus in Australia. These bring the number of eastern Australian species to twenty-six. Distributions of three other species are extended northwards into Queensland. A key to all eastern Australian species is provided.

Abstract Objectives To establish packed cell volume (PCV) ranges for non‐pregnant, pregnant and post‐partum bitches from day 10 of proestrus, investigating any relationship with parity and litter size. Methods This prospective cohort study used 37 healthy breeding bitches to examine PCV counts from routine blood samples collected every 4 weeks, from day 10 of proestrus, as part of routine PCV monitoring. Results For pregnant (n = 19) and non‐pregnant (n = 18) bitches, PCV decreased until week 8 (corresponding to 8.5 ± 1.1 days before whelping for pregnant bitches) and recovered by 16–20 weeks after the initial sample; bitches that whelped average and large litters showed greater declines. PCV began to recover sooner for bitches that had previously whelped one or two litters compared to bitches that had previously whelped three or more litters. There was a significant three‐way interaction between time after the onset of proestrus, litter size and the number of previous litters which demonstrated that the large decrease in PCV for bitches that had previously whelped three or more litters only occurred in bitches that were expecting an average or large sized litter. Clinical Significance Chronological variation in PCV for pregnant and non‐pregnant bitches was established during the reproductive cycle. There was no evidence to suggest that routine PCV measurement for normal, healthy bitches would be beneficial. However, knowledge from this study may be useful when deciding whether to prospectively monitor a bitch where there is a history of previous pregnancy‐related anaemia, when performing a caesarean section due to the anticipated blood loss during surgery, or when examining blood profiles for post‐litter bitches.

Abstract Introduction Oral administration of zeaxanthin (Zx) 20 mg daily in patients with unilateral neovascular age-related macular degeneration (nAMD) treated with triple therapy (photodynamic therapy/intravitreal bevacizumab/intravitreal dexamethasone) reduced fellow-eye 2-year nAMD incidence from 23 to 6% (p = 0.02) in a prior clinical trial. We questioned the long-term benefit and thus analyzed case–control 5-year patient data of trial participants and additional participants with 5-year follow-up, also performing cost-utility and cost–benefit analyses. Methods Consecutive, unilateral nAMD patient outcomes for those taking 20 mg Zx supplementation orally for ≥ 5 years were compared with the Comparison of AMD Treatments Trials (CATT) 5-year historical controls for fellow-eye nAMD conversion. Eleven-year mean life expectancy, cost-utility and cost–benefit models were undertaken employing a 3% discount rate and 2020 US real dollars. Results Among 227 consecutive patients with nAMD/Zx-supplementation, 202 (90%) had 5-year follow-up. The fellow-eye nAMD 5-year conversion incidence using a Kaplan–Meier cumulative event estimate was 22% (49/227), versus 48% (167/348) with CATT control data (p

Abstract Cervical spinal cord injury impacts ventilatory and non‐ventilatory functions of the diaphragm muscle (DIAm) and contributes to clinical morbidity and mortality in the afflicted population. Periodically, integrated brainstem neural circuit activity drives the DIAm to generate a markedly augmented effort or sigh—which plays an important role in preventing atelectasis and thus maintaining lung function. Across species, the general pattern of DIAm efforts during a normal sigh is variable in amplitude and the extent of post‐sigh “apnea” (i.e., the post‐sigh inter‐breath interval). This post‐sigh inter‐breath interval acts as a respiratory reset, following the interruption of regular respiratory rhythm by sigh. We examined the impact of upper cervical (C2) spinal cord hemisection (C2SH) on the transdiaphragmatic pressure (Pdi) generated during sighs and the post‐sigh respiratory reset in rats. Sighs were identified in Pdi traces by their characteristic biphasic pattern. We found that C2SH results in a reduction of Pdi during both eupnea and sighs, and a decrease in the immediate post‐sigh breath interval. These results are consistent with partial removal of descending excitatory synaptic inputs to phrenic motor neurons that results from C2SH. Following cervical spinal cord injury, a reduction in the amplitude of Pdi during sighs may compromise the maintenance of normal lung function.

BackgroundLearning to cycle can be challenging for adults who did not acquire the necessary skills during childhood. Balance bikes have been used to teach children how to cycle, but it was unclear whether this approach could also be effective for adults.PurposeTo address this, a multi-phase intervention study was conducted to investigate whether healthy adults could be taught to cycle independently through the use of a balance bike.MethodsIn Phase 1, a case-control observational study was conducted in which 13 cyclists and 8 non-cyclists completed balance bike tests. Based on the findings, an 8-session intervention pre- and post-test study was conducted in Phase 2, using an 8 × 20-minute balance bike training programme to improve cycling postural stability and control. Another 11 non-cyclists completed the novel programme. The time taken to complete the balance bike tests was compared before and after the program, while their cycling confidence was recorded in each session. To assess the effectiveness of the programme, participants were invited to cycle on a pedal bike to evaluate their ability to cycle independently.ResultsThe results in Phase 1 showed that cyclists performed better on the balance bike than non-cyclists, with Bayes factor analyses providing evidence of this difference, BF01 = 0.228 in the 15 m sprint test and BF01 = 0.138 in the two-turn curved sprint test. The novel training programme in Phase 2 demonstrated remarkable effectiveness in improving their balance bike riding performance, as evidenced by the Bayes factor for completion times in the repeated measures being BF01 < 0.001. All participants were able to cycle independently with confidence after the programme.ConclusionsThis study sheds light on the idea that it's never too late for adults to learn how to ride a bike. It provides evidence that healthy adults can learn to ride independently with the help of a balance bike, a tool that's commonly used for teaching children. The study identifies five key principles for effective balance bike training in adults, including focusing on riding speed, gliding to turn, building cycling confidence, engaging high motor skills, and using a dual-task approach. Our evidence-based training programme offers a safe, enjoyable, and effective way for adults to develop the skills and confidence they need to ride, even if they've never ridden before.

Abstract Background Hearing problems (HP) in adults are common and are associated with several comorbid conditions. Its prevalence increases with age, reflecting the cumulative effect of environmental factors and genetic predisposition. Although several risk loci have been already identified, HP biology and epidemiology are still insufficiently investigated by large-scale genetic studies. Methods Leveraging the UK Biobank, the Nurses’ Health Studies (I and II), the Health Professionals Follow-up Study, and the Million Veteran Program, we conducted a comprehensive genome-wide investigation of HP in 748,668 adult participants (discovery N = 501,825; replication N = 226,043; cross-ancestry replication N = 20,800). We leveraged the GWAS findings to characterize HP polygenic architecture, exploring sex differences, polygenic risk across ancestries, tissue-specific transcriptomic regulation, cause-effect relationships with genetically correlated traits, and gene interactions with HP environmental risk factors. Results We identified 54 risk loci and demonstrated that HP polygenic risk is shared across ancestry groups. Our transcriptomic regulation analysis highlighted the potential role of the central nervous system in HP pathogenesis. The sex-stratified analyses showed several additional associations related to peripheral hormonally regulated tissues reflecting a potential role of estrogen in hearing function. This evidence was supported by the multivariate interaction analysis that showed how genes involved in brain development interact with sex, noise pollution, and tobacco smoking in relation to their HP associations. Additionally, the genetically informed causal inference analysis showed that HP is linked to many physical and mental health outcomes. Conclusions The results provide many novel insights into the biology and epidemiology of HP in adults. Our sex-specific analyses and transcriptomic associations highlighted molecular pathways that may be targeted for drug development or repurposing. Additionally, the potential causal relationships identified may support novel preventive screening programs to identify individuals at risk.

The present study investigated the microstructure and wear properties of annealed carbon steels with the addition of TiC–TiB2 ceramic nanoparticles. It was found that the presence of 0.02 wt% nanoparticles could decrease the amount of widmanstatten structure, promote the formation of finer pearlite from 33.0% to 46.3% and refine the grains from 67.0 μm to 47.5 μm. In ball-on-plate reciprocating sliding wear tests, the wear volume loss was reduced up to 27.2% while the test load of 50 N and sliding velocity of 60 mm/s were utilized. After examining the wear tracks, it was observed that the spalls, grooves, smearing, delamination, oxidative areas and wear debris were found within the worn areas. The ceramic nanoparticles increased the hardness of substrates, inhibited dislocations and plastic deformation, retarded the cracks on surfaces and sub-surfaces, and withstand the load which enhanced the anti-wear performance of annealed carbon steels. This research could provide a new method to improve the wear performance of carbon steel components.

Summary: A challenge in studying cervical epithelial cell biology at the single-cell level is that differentiated subtypes, in particular mucus-secreting goblet cells, are sensitive to disassociating enzymes making isolation of all epithelial subpopulations difficult. Here we present a protocol to dissociate epithelia from non-pregnant and pregnant mouse cervical tissue for single-cell RNA-sequencing. We describe steps for harvesting cervices, preparing cervical tissue, dissociation of cervical cells, and viability checks. We then detail library preparation, sequencing, and procedure for data analysis.For complete details on the use and execution of this protocol, please refer to Cooley et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Background Conversational artificial intelligence (chatbots and dialogue systems) is an emerging tool for tobacco cessation that has the potential to emulate personalised human support and increase engagement. We aimed to determine the effect of conversational artificial intelligence interventions with or without standard tobacco cessation interventions on tobacco cessation outcomes among adults who smoke, compared to no intervention, placebo intervention or an active comparator. Methods A comprehensive search of six databases was completed in June 2022. Eligible studies included randomised controlled trials published since 2005. The primary outcome was sustained tobacco abstinence, self-reported and/or biochemically validated, for at least 6 months. Secondary outcomes included point-prevalence abstinence and sustained abstinence of less than 6 months. Two authors independently extracted data on cessation outcomes and completed the risk of bias assessment. Random effects meta-analysis was conducted. Results From 819 studies, five randomised controlled trials met inclusion criteria (combined sample size n = 58,796). All studies differed in setting, methodology, intervention, participants and end-points. Interventions included chatbots embedded in multi- and single-component smartphone apps ( n = 3), a social media-based ( n = 1) chatbot, and an internet-based avatar ( n = 1). Random effects meta-analysis of three studies found participants in the conversational artificial intelligence enhanced intervention were significantly more likely to quit smoking at 6-month follow-up compared to control group participants (RR = 1.29, 95% CI (1.13, 1.46), p

Abstract Impaired autophagy, a cellular digestion process that eliminates proteins and damaged organelles, has been implicated in neurodegenerative diseases, including motor neuron disorders. Motor neuron targeted upregulation of autophagy may serve as a promising therapeutic approach. Lanthionine ketenamine (LK), an amino acid metabolite found in mammalian brain tissue, activates autophagy in neuronal cell lines. We hypothesized that analogs of LK can be targeted to motor neurons using nanoparticles to improve autophagy flux. Using a mouse motor neuron-like hybrid cell line (NSC-34), we tested the effect of three different LK analogs on autophagy modulation, either alone or loaded in nanoparticles. For fluorescence visualization of autophagy flux, we used a mCherry-GFP-LC3 plasmid reporter. We also evaluated protein expression changes in LC3-II/LC3-I ratio obtained by western blot, as well as presence of autophagic vacuoles per cell obtained by electron microscopy. Delivering LK analogs with targeted nanoparticles significantly enhanced autophagy flux in differentiated motor neuron-like cells compared to LK analogs alone, suggesting the need of a delivery vehicle to enhance their efficacy. In conclusion, LK analogs loaded in nanoparticles targeting motor neurons constitute a promising treatment option to induce autophagy flux, which may serve to mitigate motor neuron degeneration/loss and preserve motor function in motor neuron disease.

Oncolytic viruses (OV) promote anti-tumour responses through the initiation of immunogenic cancer cell death which activates the host’s systemic anti-tumour immunity. We have previously shown that intravenously administered HSV1716 is an effective treatment for mammary cancer. However, intravenous administration of a virus has the potential to result in neutralization and sequestration of the virus which may reduce efficacy. Here, we show that the oncolytic virus HSV1716 can be administered within a cellular carrier (macrophages). PyMT and 4T1 murine mammary cancer cell lines were implanted into immuno-competent murine models (orthotopic primary, early metastatic and brain metastasis models). HSV1716 or macrophages armed with HSV1716 (M-HSV1716) were administered intravenously, and tumour size was quantified using caliper measurement or bioluminescence imaging. Administration of M-HSV1716 led to tumour shrinkage and increased the survival of animals. Furthermore, these results were achieved with a 100-fold lower viral load, which has the potential for decreased toxicity. Our results demonstrate that M-HSV1716 is associated with activity against murine mammary cancers and provides an alternative platform for the systemic delivery of OV.

Summary: Resting metabolic rate (RMR) adaptation occurs during obesity and is hypothesized to contribute to failed weight management. Angiotensin II (Ang-II) type 1 (AT1A) receptors in Agouti-related peptide (AgRP) neurons contribute to the integrative control of RMR, and deletion of AT1A from AgRP neurons causes RMR adaptation. Extracellular patch-clamp recordings identify distinct cellular responses of individual AgRP neurons from lean mice to Ang-II: no response, inhibition via AT1A and Gαi, or stimulation via Ang-II type 2 (AT2) receptors and Gαq. Following diet-induced obesity, a subset of Ang-II/AT1A-inhibited AgRP neurons undergo a spontaneous G-protein “signal switch,” whereby AT1A stop inhibiting the cell via Gαi and instead begin stimulating the cell via Gαq. DREADD-mediated activation of Gαi, but not Gαq, in AT1A-expressing AgRP cells stimulates RMR in lean and obese mice. Thus, loss of AT1A-Gαi coupling within the AT1A-expressing AgRP neuron subtype represents a molecular mechanism contributing to RMR adaptation.

Despite ground-breaking genetic studies that have identified thousands of risk variants for developmental diseases, how these variants lead to molecular and cellular phenotypes remains a gap in knowledge. Many of these variants are non-coding and occur at enhancers, which orchestrate key regulatory programs during development. The prevailing paradigm is that non-coding variants alter the activity of enhancers, impacting gene expression programs, and ultimately contributing to disease risk. A key obstacle to progress is the systematic functional characterization of non-coding variants at scale, especially since enhancer activity is highly specific to cell type and developmental stage. Here, we review the foundational studies of enhancers in developmental disease and current genomic approaches to functionally characterize developmental enhancers and their variants at scale. In the coming decade, we anticipate systematic enhancer perturbation studies to link non-coding variants to molecular mechanisms, changes in cell state, and disease phenotypes.

Using high-resolution 4D imaging, Kidokoro et al. identify two key cell behaviors shaping the early heart tube. They further show that left-specific Nodal signaling promotes these cell behaviors, thereby driving tissue-scale left-right asymmetry.

The present research utilized the resistance spot welding process on stainless steel and low carbon steel. The diameters of joint and nugget, microstructure and shearing strength of joints were examined. When increasing the welding current, welding time or clamping force, the diameters of welding joints and nuggets became larger due to the higher heat input and normal load. The shearing strength of joints increased and then decreased with increasing welding current or welding time and decreased with increasing clamping force. For the metallurgical evaluation, it was found that there were five regions around the joint, which consisted of the nugget (austenite and martensite with dendritic structures), coarse heat-affected zone (martensite and pro-eutectoid ferrite), fine heat-affected zone (ferrite and pearlite), base material (ferrite and pearlite) for low carbon steel, and base material (austenite) for stainless steel. In orthogonal tests, it was found that the welding current had the most significant influences on shearing strength, and then welding time and clamping force. Finally, an artificial neural network was established to predict the shearing strength of joints under various welding conditions. The results showed that the shearing strengths predicted by the artificial neural network had good correlation with experimental values. A shearing strength higher than 10000 N could be achieved when welding current was higher than 12.5 kA and welding time was longer than 85 ms. The artificial neural network model has applicable values for predicting the shearing strengths of joints between dissimilar steel plates.

Enhancers orchestrate gene expression programs that drive multicellular development and lineage commitment. Thus, genetic variants at enhancers are thought to contribute to developmental diseases by altering cell fate commitment. However, while many variant-containing enhancers have been identified, studies to endogenously test the impact of these enhancers on lineage commitment have been lacking. We perform a single-cell CRISPRi screen to assess the endogenous roles of 25 enhancers and putative cardiac target genes implicated in genetic studies of congenital heart defects (CHDs). We identify 16 enhancers whose repression leads to deficient differentiation of human cardiomyocytes (CMs). A focused CRISPRi validation screen shows that repression of TBX5 enhancers delays the transcriptional switch from mid- to late-stage CM states. Endogenous genetic deletions of two TBX5 enhancers phenocopy epigenetic perturbations. Together, these results identify critical enhancers of cardiac development and suggest that misregulation of these enhancers could contribute to cardiac defects in human patients.

Abstract Duchenne muscular dystrophy (DMD) is characterized by progressive muscle degeneration. Two important deleterious features are a Ca2+ dysregulation linked to Ca2+ influxes associated with ryanodine receptor hyperactivation, and a muscular nicotinamide adenine dinucleotide (NAD+) deficit. Here, we identified that deletion in mdx mice of CD38, a NAD+ glycohydrolase‐producing modulators of Ca2+ signaling, led to a fully restored heart function and structure, with skeletal muscle performance improvements, associated with a reduction in inflammation and senescence markers. Muscle NAD+ levels were also fully restored, while the levels of the two main products of CD38, nicotinamide and ADP‐ribose, were reduced, in heart, diaphragm, and limb. In cardiomyocytes from mdx/CD38−/− mice, the pathological spontaneous Ca2+ activity was reduced, as well as in myotubes from DMD patients treated with isatuximab (SARCLISA®) a monoclonal anti‐CD38 antibody. Finally, treatment of mdx and utrophin–dystrophin‐deficient (mdx/utr−/−) mice with CD38 inhibitors resulted in improved skeletal muscle performances. Thus, we demonstrate that CD38 actively contributes to DMD physiopathology. We propose that a selective anti‐CD38 therapeutic intervention could be highly relevant to develop for DMD patients.

Abstract Background Single-cell CRISPR screens are powerful tools to understand genome function by linking genetic perturbations to transcriptome-wide phenotypes. However, since few cells can be affordably sequenced in these screens, biased sampling of cells could affect data interpretation. One potential source of biased sampling is clonal cell expansion. Results Here, we identify clonal cells in single cell screens using multiplexed sgRNAs as barcodes. We find that the cells in each clone share transcriptional similarities and bear segmental copy number changes. These analyses suggest that clones are genetically distinct. Finally, we show that the transcriptional similarities of clonally expanded cells contribute to false positives in single-cell CRISPR screens. Conclusions Experimental conditions that reduce clonal expansion or computational filtering of clonal cells will improve the reliability of single-cell CRISPR screens.

During pyroptosis, gasdermin D (GSDMD) forms plasma membrane pores that initiate cell lysis. Here, the authors develop optogenetically activatable human GSDMD to assess GSDMD pore behavior and show that they are dynamic and can close, which can be a pyroptosis regulatory mechanism.

The present research investigated the effects of welding current and welding time on the shearing strength of resistance spot-welded joints of low carbon steel and stainless steel plates. The macro characteristics, microstructure and micro-hardness of welded joints were also examined. It was found that the welded joints produced with a welding current of 10 kA and welding time of 80 ms showed the highest shearing strength. The diameter of welded joints and nuggets increased with increasing welding current and welding time. For evaluation of the microstructure, different regions including the nugget, coarse heat-affected zone, fine heat-affected zone and base materials were observed within the welded joints. Also, the nugget and coarse heat-affected zone showed higher hardness than the fine heat-affected zone and base materials due to the presence of martensite. The findings obtained in this research are applicable in the development of resistance spot welding processes for joining dissimilar steel plates.

Comprehensive cis-regulatory landscapes are essential for accurate enhancer prediction and disease variant mapping. Although cis-regulatory element (CRE) resources exist for most tissues and organs, many rare — yet functionally important — cell types remain overlooked. Despite representing only a small fraction of the heart’s cellular biomass, the cardiac conduction system (CCS) unfailingly coordinates every life-sustaining heartbeat. To globally profile the mouse CCS cis-regulatory landscape, we genetically tagged CCS component–specific nuclei for comprehensive assay for transposase-accessible chromatin–sequencing (ATAC-Seq) analysis. Thus, we established a global CCS-enriched CRE database, referred to as CCS-ATAC, as a key resource for studying CCS-wide and component-specific regulatory functions. Using transcription factor (TF) motifs to construct CCS component–specific gene regulatory networks (GRNs), we identified and independently confirmed several specific TF sub-networks. Highlighting the functional importance of CCS-ATAC, we also validated numerous CCS-enriched enhancer elements and suggested gene targets based on CCS single–cell RNA-Seq data. Furthermore, we leveraged CCS-ATAC to improve annotation of existing human variants related to cardiac rhythm and nominated a potential enhancer-target pair that was dysregulated by a specific SNP. Collectively, our results established a CCS-regulatory compendium, identified novel CCS enhancer elements, and illuminated potential functional associations between human genomic variants and CCS component–specific CREs.

Summary: The cervical epithelium undergoes changes in proliferation, differentiation, and function that are critical to ensure fertility and maintain pregnancy. Here, we identify cervical epithelial subtypes in non-pregnant, pregnant, and in labor mice using single-cell transcriptome and spatial analysis. We identify heterogeneous subpopulations of epithelia displaying spatial and temporal specificity. Notably in pregnancy, two goblet cell subtypes are present in the most luminal layers with one goblet population expanding earlier in pregnancy than the other goblet population. The goblet populations express novel protective factors and distinct mucosal networks. Single-cell analysis in a model of cervical epithelial barrier disruption indicates untimely basal cell proliferation precedes the expansion of goblet cells with diminished mucosal integrity. These data demonstrate how the cervical epithelium undergoes continuous remodeling to maintain dynamic states of homeostasis in pregnancy and labor, and provide a framework to understand perturbations in epithelial health that increase the risk of premature birth.

Abstract Diaphragm muscle (DIAm) motor units comprise a phrenic motor neuron (PhMN), the phrenic nerve and the muscle fibers innervated, with the size of PhMNs and axons characteristic of motor unit type. Smaller PhMNs and their axons comprise slow (type S) and fatigue‐resistant (type FR) DIAm motor units, while larger PhMNs and their axons comprise more fatigable (type FF) motor units. With aging, we have shown a loss of larger PhMNs, consistent with selective atrophy of type IIx/IIb DIAm fibers and reduced maximum DIAm force. In the present study, we hypothesized that with aging there is a loss of larger myelinated phrenic α motor axons. Female and male young (6 months) and old (24 months) Fischer 344 rats were studied. PhMNs were retrogradely labeled by intrapleural injection of 488‐conjugated CTB. The phrenic nerves were excised ~1 cm from the DIAm insertion and mounted in resin, and phrenic α motor axons were delineated based on size (i.e., >4 μm diameters). In older rats, the number of larger PhMNs and larger phrenic α motor axons were reduced. There were no differences in non‐α axons. In addition, there was evidence of demyelination of larger phrenic α motor axons in older rats. Together, these findings are consistent with the selective age‐related vulnerability of larger PhMNs and denervation of type FF motor units, which may underlie DIAm sarcopenia.

Abstract Background Human cytomegalovirus (HCMV) resides latently in cells of the myeloid compartment, including CD34+ hematopoietic progenitor cells and circulating monocytes. Healthy hosts maintain the virus latently, and this infection is, for the most part, asymptomatic. However, given the proper external cues, HCMV reactivates from latency, at which point the virus disseminates, causing disease. The viral and cellular factors dictating the balance between these phases of infection are incompletely understood, though a large body of literature support a role for viral-mediated manipulation of host cell signaling. Main body To establish and maintain latency, HCMV has evolved various means by which it usurps host cell factors to alter the cellular environment to its own advantage, including altering host cell signaling cascades. As early as virus entry into myeloid cells, HCMV usurps cellular signaling to change the cellular milieu, and this regulation includes upregulation, as well as downregulation, of different signaling cascades. Indeed, given proper reactivation cues, this signaling is again altered to allow for transactivation of viral lytic genes. Conclusions HCMV modulation of host cell signaling is not binary, and many of the cellular pathways altered are finely regulated, wherein the slightest modification imparts profound changes to the cellular milieu. It is also evident that viral-mediated cell signaling differs not only between these phases of infection, but also is myeloid cell type specific. Nonetheless, understanding the exact pathways and the means by which HCMV mediates them will undoubtedly provide novel targets for therapeutic intervention.

Background We investigated the longitudinal association of herpes zoster (HZ), commonly known as “shingles,” and long‐term risk of stroke or coronary heart disease (CHD) among participants in 3 large US cohorts, the NHS (Nurses' Health Study), NHS II (Nurses' Health Study II), and HPFS (Health Professionals Follow‐Up Study). Methods and Results Participants were 79 658 women in the NHS (2000–2016), 93 932 women in the NHS II (2001–2017), and 31 440 men in the HPFS (2004–2016), without prior stroke or CHD. Information on HZ, stroke, and CHD was collected on biennial questionnaires and confirmed by medical record review. Cox proportional hazards regression models were used to estimate multivariable‐adjusted hazard ratios for stroke and for CHD according to years since HZ compared with never HZ. During >2 million person‐years of follow‐up, 3603 incident stroke and 8620 incident CHD cases were documented. History of HZ was significantly and independently associated with higher long‐term risk of stroke and CHD. In pooled analyses, compared with individuals with no history of HZ, the multivariable‐adjusted hazard ratios (95% CIs) for stroke were 1.05 (0.88–1.25) among those with 1 to 4 years since HZ, 1.38 (1.10–1.74) for among those with 5 to 8 years since HZ, 1.28 (1.03–1.59) among those with for 9 to 12 years since HZ, and 1.19 (0.90–1.56) among those with ≥13 years since HZ. For CHD, the corresponding multivariable‐adjusted hazard ratios (95% CIs) were 1.13 (1.01–1.27) for 1 to 4 years, 1.16 (1.02–1.32) for 5 to 8 years, 1.25 (1.07–1.46) for 9 to 12 years, and 1.00 (0.83–1.21) for ≥13 years. Conclusions HZ is associated with higher long‐term risk of a major cardiovascular event. These findings suggest there are long‐term implications of HZ and underscore the importance of prevention.

The description in 1891 of the sea pen genus Gyrophyllum Studer, 1891 and also the type species G. hirondellei Studer, 1891 was based on a single colony collected in the Azores Archipelago. During the 19th and 20th centuries, the family placement of this genus became controversial as the set of morphological features present in Gyrophyllum could justify its assignation to both the families Pennatulidae Ehrenberg, 1834 and Pteroeididae Kölliker, 1880. Deliberations over this intermediate set of characters finally ended in the reunification of the genera and species of both families under Pennatulidae by principle of priority. The use of molecular sources of information based on a series of sequencing techniques presents a different but promising phylogenetic scenario in order to go further in the understanding of pennatulacean systematics. In this paper, a complementary morphological and molecular study (multiloci sequences with three mitochondrial and one nuclear markers) based mainly on newly collected material is carried out. This study re-confirms from a molecular point of view previously published results that indicate the position of Gyrophyllum as being distant from Pennatula Linnaeus, 1758 and Pteroeides Herklots, 1858 (type genera of the families Pennatulidae and Pteroeididae, respectively). This fact together with the results of a detailed morphological examination strongly supports the placement of the enigmatic genus Gyrophyllum in a separate family: Gyrophyllidae fam. nov. and resolves the nomenclatural uncertainty at family level for this genus. Moreover, the characters previously considered useful in the distinction of the two currently recognised species G. hirondellei in the Atlantic and G. sibogae Hickson, 1916 in the Indo-western Pacific are revisited.