Honey bees and social wasps reach convergent architectural solutions to nest-building problems.

The hexagonal cells built by honey bees and social wasps are an example of adaptive architecture; hexagons minimize material use, while maximizing storage space and structural stability. Hexagon building evolved independently in the bees and wasps, but in some species of both groups, the hexagonal cells are size dimorphic-small worker cells and large reproductive cells-which forces the builders to join differently sized hexagons together. This inherent tiling problem creates a unique opportunity to investigate how similar architectural challenges are solved across independent evolutionary origins. We investigated how 5 honey bee and 5 wasp species solved this problem by extracting per-cell metrics from 22,745 cells. Here, we show that all species used the same building techniques: intermediate-sized cells and pairs of non-hexagonal cells, which increase in frequency with increasing size dimorphism. We then derive a simple geometric model that explains and predicts the observed pairing of non-hexagonal cells and their rate of occurrence. Our results show that despite different building materials, comb configurations, and 179 million years of independent evolution, honey bees and social wasps have converged on the same solutions for the same architectural problems, thereby revealing fundamental building properties and evolutionary convergence in construction behavior.

A Large-Scale Study of the Relationship Between Degree and Type of Hearing Loss and Recognition of Speech in Quiet and Noise.

OBJECTIVES: Understanding speech in noise (SIN) is the dominant complaint of individuals with hearing loss. For decades, the default test of speech perception in routine audiologic assessment has been monosyllabic word recognition in quiet (WRQ), which does not directly address patient concerns, leading some to advocate that measures of SIN should be integrated into routine practice. However, very little is known with regard to how SIN abilities are affected by different types of hearing loss. Here, we examine performance on clinical measures of WRQ and SIN in a large patient base consisting of a variety of hearing loss types, including conductive (CHL), mixed (MHL), and sensorineural (SNHL) losses. DESIGN: In a retrospective study, we examined data from 5593 patients (51% female) who underwent audiometric assessment at the Stanford Ear Institute. All individuals completed pure-tone audiometry, and speech perception testing of monaural WRQ, and monaural QuickSIN. Patient ages ranged from 18 to 104 years (average = 57). The average age in years for the different classifications of hearing loss was 51.1 (NH), 48.5 (CHL), 64.2 (MHL), and 68.5 (SNHL), respectively. Generalized linear mixed-effect models and quartile regression were used to determine the relationship between hearing loss type and severity for the different speech-recognition outcome measures. RESULTS: Patients with CHL had similar performance to patients with normal hearing on both WRQ and QuickSIN, regardless of the hearing loss severity. In patients with MHL or SNHL, WRQ scores remained largely excellent with increasing hearing loss until the loss was moderately severe or worse. In contrast, QuickSIN signal to noise ratio (SNR) losses showed an orderly systematic decrease as the degree of hearing loss became more severe. This effect scaled with the data, with threshold-QuickSIN relationships absent for CHL, and becoming increasingly stronger for MHL and strongest in patients with SNHL. However, the variability in these data suggests that only 57% of the variance in WRQ scores, and 50% of the variance in QuickSIN SNR losses, could be accounted for by the audiometric thresholds. Patients who would not be differentiated by WRQ scores are shown to be potentially differentiable by SIN scores. CONCLUSIONS: In this data set, conductive hearing loss had little effect on WRQ scores or QuickSIN SNR losses. However, for patients with MHL or SNHL, speech perception abilities decreased as the severity of the hearing loss increased. In these data, QuickSIN SNR losses showed deficits in performance with degrees of hearing loss that yielded largely excellent WRQ scores. However, the considerable variability in the data suggests that even after classifying patients according to their type of hearing loss, hearing thresholds only account for a portion of the variance in speech perception abilities, particularly in noise. These results are consistent with the idea that variables such as cochlear health and aging add explanatory power over audibility alone.

Speech-in-Noise Assessment in the Routine Audiologic Test Battery: Relationship to Perceived Auditory Disability.

OBJECTIVES: Self-assessment of perceived communication difficulty has been used in clinical and research practices for decades. Such questionnaires routinely assess the perceived ability of an individual to understand speech, particularly in background noise. Despite the emphasis on perceived performance in noise, speech recognition in routine audiologic practice is measured by word recognition in quiet (WRQ). Moreover, surprisingly little data exist that compare speech understanding in noise (SIN) abilities to perceived communication difficulty. Here, we address these issues by examining audiometric thresholds, WRQ scores, QuickSIN signal to noise ratio (SNR) loss, and perceived auditory disability as measured by the five questions on the Speech Spatial Questionnaire-12 (SSQ12) devoted to speech understanding (SSQ12-Speech5). DESIGN: We examined data from 1633 patients who underwent audiometric assessment at the Stanford Ear Institute. All individuals completed the SSQ12 questionnaire, pure-tone audiometry, and speech assessment consisting of ear-specific WRQ, and ear-specific QuickSIN. Only individuals with hearing threshold asymmetries ≤10 dB HL in their high-frequency pure-tone average (HFPTA) were included. Our primary objectives were to (1) examine the relationship between audiometric variables and the SSQ12-Speech5 scores, (2) determine the amount of variance in the SSQ12-Speech5 scores which could be predicted from audiometric variables, and (3) predict which patients were likely to report greater perceived auditory disability according to the SSQ12-Speech5. RESULTS: Performance on the SSQ12-Speech5 indicated greater perceived auditory disability with more severe degrees of hearing loss and greater QuickSIN SNR loss. Degree of hearing loss and QuickSIN SNR loss were found to account for modest but significant variance in SSQ12-Speech5 scores after accounting for age. In contrast, WRQ scores did not significantly contribute to the predictive power of the model. Degree of hearing loss and QuickSIN SNR loss were also found to have moderate diagnostic accuracy for determining which patients were likely to report SSQ12-Speech5 scores indicating greater perceived auditory disability. CONCLUSIONS: Taken together, these data indicate that audiometric factors including degree of hearing loss (i.e., HFPTA) and QuickSIN SNR loss are predictive of SSQ12-Speech5 scores, though notable variance remains unaccounted for after considering these factors. HFPTA and QuickSIN SNR loss-but not WRQ scores-accounted for a significant amount of variance in SSQ12-Speech5 scores and were largely effective at predicting which patients are likely to report greater perceived auditory disability on the SSQ12-Speech5. This provides further evidence for the notion that speech-in-noise measures have greater clinical utility than WRQ in most instances as they relate more closely to measures of perceived auditory disability.

Imperfect comb construction reveals the architectural abilities of honeybees.

Honeybees are renowned for their perfectly hexagonal honeycomb, hailed as the pinnacle of biological architecture for its ability to maximize storage area while minimizing building material. However, in natural nests, workers must regularly transition between different cell sizes, merge inconsistent combs, and optimize construction in constrained geometries. These spatial obstacles pose challenges to workers building perfect hexagons, but it is unknown to what extent workers act as architects versus simple automatons during these irregular building scenarios. Using automated image analysis to extract the irregularities in natural comb building, we show that some building configurations are more difficult for the bees than others, and that workers overcome these challenges using a combination of building techniques, such as: intermediate-sized cells, regular motifs of irregular shapes, and gradual modifications of cell tilt. Remarkably, by anticipating these building challenges, workers achieve high-quality merges using limited local sensing, on par with analytical models that require global optimization. Unlike automatons building perfectly replicated hexagons, these building irregularities showcase the active role that workers take in shaping their nest and the true architectural abilities of honeybees.

Manipulating nest architecture reveals three-dimensional building strategies and colony resilience in honeybees.

Form follows function throughout the development of an organism. This principle should apply beyond the organism to the nests they build, but empirical studies are lacking. Honeybees provide a uniquely suited system to study nest form and function throughout development because we can image the three-dimensional structure repeatedly and non-destructively. Here, we tracked nest-wide comb growth in six colonies over 45 days (control colonies) and found that colonies have a stereotypical process of development that maintains a spheroid nest shape. To experimentally test if nest structure is important for colony function, we shuffled the nests of an additional six colonies, weekly rearranging the comb positions and orientations (shuffled colonies). Surprisingly, we found no differences between control and shuffled colonies in multiple colony performance metrics-worker population, comb area, hive weight and nest temperature. However, using predictive modelling to examine how workers allocate comb to expand their nests, we show that shuffled colonies compensate for these disruptions by accounting for the three-dimensional structure to reconnect their nest. This suggests that nest architecture is more flexible than previously thought, and that superorganisms have mechanisms to compensate for drastic architectural perturbations and maintain colony function.

High dietary iodine intake may contribute to the low death rate from COVID-19 infection in Japan with activation by the lactoperoxidase system.

We draw the attention of readers and governments to the death rate from coronavirus disease 2019 in Japan, continuing as a fraction of that experienced by many other developed nations. We think this is due to the activity of the powerful, protective lactoperoxidase system (LPO) which prevents serious airborne infections. The LPO system requires iodine, which is liberally provided by the typical Japanese diet but lacking in many others. One might consider the Japanese experience an incredibly large, open-label study exhibiting the preventative power of a high-iodine diet. We predict this favourable trend will continue for Japan because deadly variants of the severe, acute respiratory syndrome coronavirus 2 will be with us, forever.

Comments on the Discussion Forum: Oromucosal immunomodulation as clinical spectrum mitigating factor in SARS-CoV-2 infection.

The mammalian lactoperoxidase system, consisting of lactoperoxidase and the H2 O2 -producing enzyme duox, is our first line of defence against airborne microbes. This system catalyses the production of hypoiodite and hypoiodous acid in the presence of sufficient iodine. These products are highly efficient at destroying the H1N1 virus and the respiratory syncytial virus (RSV). Japan has not been affected as much as other nations during the COVID-19 pandemic (death rate about 10% of the United States), and we think this is due to a diet high in iodine. With this in mind, we suggest four actions to prevent SARS-CoV-2 infections. First, health professionals should study the preventative effect of increasing iodine in the diets of the aged, institutionalized, diabetics andsmokers. Second, the recommended daily intake (RDI) for iodine should be significantly increased, to at least double, the current RDI. Governments should encourage the use and distribution of cheap iodized salts, kelp and seaweed. Third, more research should be done around the physiology and the protective effects of the lactoperoxidase system. Finally, the degradation products of the SARS-CoV-2 viral particle by hypoiodite and hypoiodous acid should be characterized; portions of the damaged particle are likely to elicit stronger immunity and better vaccines.

Reply to the 'Comment on "Tensional homeostasis at different length scales" by J. Humphrey and C. Cyron, Soft Matter, 2022, 18, DOI: 10.1039/D1SM01151K'.

Drs Humphrey and Cyron wrote a commentary regarding our review article entitled "Tensional homeostasis at different length scales" that was published in Soft Matter, 2020, 16, 6946-6963. These authors brought up some valid concerns to which we would like to respond. Their first concern is related to our remark regarding equations that we used to describe homeostasis in blood vessels, where we stated that those equations were limited only to linearly elastic materials. We were wrong, and we agree with the authors that these equations hold for all cylindrical vessels regardless of their material properties. Their second concern is related to tensional homeostasis at the subcellular level. Drs Humphrey and Cyron disagree with our substantiated claim that tensional homeostasis breaks down at the level of focal adhesions (FAs) of a living cell. In our reply, we provided several pieces of evidence that demonstrate that tensional homeostasis depends upon FA size, FA maturity and FA force dynamics and thus, tensional homeostasis cannot hold in all FAs across a cell. In summary, we are grateful for the opportunity to reply to the commentary of Drs Humphrey and Cyron. Moreover, we are excited that this topic has become an important focus in the biomechanics and mechanobiology communities, and we feel strongly that critical feedback is necessary to move this field forward.

Integrin ß1 orchestrates the abnormal cell-matrix attachment and invasive behaviour of E-cadherin dysfunctional cells.

BACKGROUND: Tumour progression relies on the ability of cancer cells to penetrate and invade neighbouring tissues. E-cadherin loss is associated with increased cell invasion in gastric carcinoma, and germline mutations of the E-cadherin gene are causative of hereditary diffuse gastric cancer. Although E-cadherin dysfunction impacts cell-cell adhesion, cell dissemination also requires an imbalance of adhesion to the extracellular matrix (ECM). METHODS: To identify ECM components and receptors relevant for adhesion of E-cadherin dysfunctional cells, we implemented a novel ECM microarray platform coupled with molecular interaction networks. The functional role of putative candidates was determined by combining micropattern traction microscopy, protein modulation and in vivo approaches, as well as transcriptomic data of 262 gastric carcinoma samples, retrieved from the cancer genome atlas (TCGA). RESULTS: Here, we show that E-cadherin mutations induce an abnormal interplay of cells with specific components of the ECM, which encompasses increased traction forces and Integrin ß1 activation. Integrin ß1 synergizes with E-cadherin dysfunction, promoting cell scattering and invasion. The significance of the E-cadherin-Integrin ß1 crosstalk was validated in Drosophila models and found to be consistent with evidence from human gastric carcinomas, where increased tumour grade and poor survival are associated with low E-cadherin and high Integrin ß1 levels. CONCLUSIONS: Integrin ß1 is a key mediator of invasion in carcinomas with E-cadherin impairment and should be regarded as a biomarker of poor prognosis in gastric cancer.

Mechanisms of aortic carboxypeptidase-like protein secretion and identification of an intracellularly retained variant associated with Ehlers-Danlos syndrome.

Aortic carboxypeptidase-like protein (ACLP) is a collagen-binding extracellular matrix protein that has important roles in wound healing and fibrosis. ACLP contains thrombospondin repeats, a collagen-binding discoidin domain, and a catalytically inactive metallocarboxypeptidase domain. Recently, mutations in the ACLP-encoding gene, AE-binding protein 1 (AEBP1), have been discovered, leading to the identification of a new variant of Ehlers-Danlos syndrome causing connective tissue disruptions in multiple organs. Currently, little is known about the mechanisms of ACLP secretion or the role of post-translational modifications in these processes. We show here that the secreted form of ACLP contains N-linked glycosylation and that inhibition of glycosylation results in its intracellular retention. Using site-directed mutagenesis, we determined that glycosylation of Asn-471 and Asn-1030 is necessary for ACLP secretion and identified a specific N-terminal proteolytic ACLP fragment. To determine the contribution of secreted ACLP to extracellular matrix mechanical properties, we generated and mechanically tested wet-spun collagen ACLP composite fibers, finding that ACLP enhances the modulus (or stiffness), toughness, and tensile strength of the fibers. Some AEBP1 mutations were null alleles, whereas others resulted in expressed proteins. We tested the hypothesis that a recently discovered 40-amino acid mutation and insertion in the ACLP discoidin domain regulates collagen binding and assembly. Interestingly, we found that this protein variant is retained intracellularly and induces endoplasmic reticulum stress identified with an XBP1-based endoplasmic reticulum stress reporter. Our findings highlight the importance of N-linked glycosylation of ACLP for its secretion and contribute to our understanding of ACLP-dependent disease pathologies.

Individual Variability in Recalibrating to Spectrally Shifted Speech: Implications for Cochlear Implants.

OBJECTIVES: Cochlear implant (CI) recipients are at a severe disadvantage compared with normal-hearing listeners in distinguishing consonants that differ by place of articulation because the key relevant spectral differences are degraded by the implant. One component of that degradation is the upward shifting of spectral energy that occurs with a shallow insertion depth of a CI. The present study aimed to systematically measure the effects of spectral shifting on word recognition and phoneme categorization by specifically controlling the amount of shifting and using stimuli whose identification specifically depends on perceiving frequency cues. We hypothesized that listeners would be biased toward perceiving phonemes that contain higher-frequency components because of the upward frequency shift and that intelligibility would decrease as spectral shifting increased. DESIGN: Normal-hearing listeners (n = 15) heard sine wave-vocoded speech with simulated upward frequency shifts of 0, 2, 4, and 6 mm of cochlear space to simulate shallow CI insertion depth. Stimuli included monosyllabic words and /b/-/d/ and /∫/-/s/ continua that varied systematically by formant frequency transitions or frication noise spectral peaks, respectively. Recalibration to spectral shifting was operationally defined as shifting perceptual acoustic-phonetic mapping commensurate with the spectral shift. In other words, adjusting frequency expectations for both phonemes upward so that there is still a perceptual distinction, rather than hearing all upward-shifted phonemes as the higher-frequency member of the pair. RESULTS: For moderate amounts of spectral shifting, group data suggested a general "halfway" recalibration to spectral shifting, but individual data suggested a notably different conclusion: half of the listeners were able to recalibrate fully, while the other halves of the listeners were utterly unable to categorize shifted speech with any reliability. There were no participants who demonstrated a pattern intermediate to these two extremes. Intelligibility of words decreased with greater amounts of spectral shifting, also showing loose clusters of better- and poorer-performing listeners. Phonetic analysis of word errors revealed certain cues were more susceptible to being compromised due to a frequency shift (place and manner of articulation), while voicing was robust to spectral shifting. CONCLUSIONS: Shifting the frequency spectrum of speech has systematic effects that are in line with known properties of speech acoustics, but the ensuing difficulties cannot be predicted based on tonotopic mismatch alone. Difficulties are subject to substantial individual differences in the capacity to adjust acoustic-phonetic mapping. These results help to explain why speech recognition in CI listeners cannot be fully predicted by peripheral factors like electrode placement and spectral resolution; even among listeners with functionally equivalent auditory input, there is an additional factor of simply being able or unable to flexibly adjust acoustic-phonetic mapping. This individual variability could motivate precise treatment approaches guided by an individual's relative reliance on wideband frequency representation (even if it is mismatched) or limited frequency coverage whose tonotopy is preserved.

A scientist like me: demographic analysis of biology textbooks reveals both progress and long-term lags.

Textbooks shape teaching and learning in introductory biology and highlight scientists as potential role models who are responsible for significant discoveries. We explore a potential demographic mismatch between the scientists featured in textbooks and the students who use textbooks to learn core concepts in biology. We conducted a demographic analysis by extracting hundreds of human names from common biology textbooks and assessing the binary gender and race of featured scientists. We found that the most common scientists featured in textbooks are white men. However, women and scientists of colour are increasingly represented in contemporary scientific discoveries. In fact, the proportion of women highlighted in textbooks has increased in lockstep with the proportion of women in the field, indicating that textbooks are matching a changing demographic landscape. Despite these gains, the scientists portrayed in textbooks are not representative of their target audience-the student population. Overall, very few scientists of colour were highlighted, and projections suggest it could take multiple centuries at current rates before we reach inclusive representation. We call upon textbook publishers to expand upon the scientists they highlight to reflect the diverse population of learners in biology.

Tensional homeostasis at different length scales.

Tensional homeostasis is a phenomenon of fundamental importance in mechanobiology. It refers to the ability of organs, tissues, and cells to respond to external disturbances by maintaining a homeostatic (set point) level of mechanical stress (tension). It is well documented that breakdown in tensional homeostasis is the hallmark of progression of diseases, including cancer and atherosclerosis. In this review, we surveyed quantitative studies of tensional homeostasis with the goal of providing characterization of this phenomenon across a broad range of length scales, from the organ level to the subcellular level. We considered both static and dynamics approaches that have been used in studies of this phenomenon. Results that we found in the literature and that we obtained from our own investigations suggest that tensional homeostasis is an emergent phenomenon driven by collective rheostatic mechanisms associated with focal adhesions, and by a collective action of cells in multicellular forms, whose impact on tensional homeostasis is cell type-dependent and cell microenvironment-dependent. Additionally, the finding that cadherins, adhesion molecules that are important for formation of cell-cell junctions, promote tensional homeostasis even in single cells, demonstrates their relevance as a signaling moiety.

On Force and Form: Mechano-Biochemical Regulation of Extracellular Matrix.

The extracellular matrix is well-known for its structural role in supporting cells and tissues, and its important biochemical role in providing signals to cells has increasingly become apparent. These structural and biochemical roles are closely coupled through mechanical forces: the biochemistry of the extracellular matrix determines its mechanical properties, mechanical forces control release or display of biochemical signals from the extracellular matrix, and the mechanical properties of the matrix in turn influence the mechanical set point at which signals are sent. In this Perspective, we explain how the extracellular matrix is regulated by strain and mechanical forces. We show the impact of biochemistry and mechanical forces on in vivo assembly of extracellular matrix and illustrate how matrix can be generated in vitro using a variety of methods. We cover how the matrix can be characterized in terms of mechanics, composition, and conformation to determine its properties and to predict interactions. Finally, we explore how extracellular matrix remodeling, ligand binding, and hemostasis are regulated by mechanical forces. These recently discovered mechano-biochemical interactions have important functions in wound healing and disease progression. It is likely that mechanically altered extracellular matrix interactions are a commonly recurring theme, but due to limited tools to generate extracellular matrix fibers in vitro and lack of high-throughput methods to detect these interactions, it is hypothesized that many of these interactions have yet to be discovered.

Single-platelet nanomechanics measured by high-throughput cytometry.

Haemostasis occurs at sites of vascular injury, where flowing blood forms a clot, a dynamic and heterogeneous fibrin-based biomaterial. Paramount in the clot's capability to stem haemorrhage are its changing mechanical properties, the major drivers of which are the contractile forces exerted by platelets against the fibrin scaffold. However, how platelets transduce microenvironmental cues to mediate contraction and alter clot mechanics is unknown. This is clinically relevant, as overly softened and stiffened clots are associated with bleeding and thrombotic disorders. Here, we report a high-throughput hydrogel-based platelet-contraction cytometer that quantifies single-platelet contraction forces in different clot microenvironments. We also show that platelets, via the Rho/ROCK pathway, synergistically couple mechanical and biochemical inputs to mediate contraction. Moreover, highly contractile platelet subpopulations present in healthy controls are conspicuously absent in a subset of patients with undiagnosed bleeding disorders, and therefore may function as a clinical diagnostic biophysical biomarker.

The cues of colony size: how honey bees sense that their colony is large enough to begin to invest in reproduction.

As organisms develop, they first invest resources in survival and growth, but after reaching a certain condition they start to also invest in reproduction. Likewise, superorganisms, such as honey bee colonies, first invest in survival and growth, and later commit resources to reproduction once the number of workers in the colony surpasses a reproductive threshold. The first form of reproductive investment for a honey bee colony is the building of beeswax comb made of special large cells used for rearing males (drones). How do the workers sense that their colony is large enough to start building this 'drone comb'? To address this question, we experimentally increased three possible cues of colony size - worker density, volatile pheromone concentration and nest temperature - and looked for effects on the bees' comb construction. Only the colonies that experienced increased worker density were stimulated to build a higher proportion of drone comb. We then monitored and quantified potential cues in small and large colonies, to determine which cues change with colony size. We found that workers in large colonies, relative to small ones, have increased contact rates, spend more time active and experience less variable worker density. Whereas unicellular and multicellular organisms use mainly chemical cues to sense their sizes, our results suggest that at least one superorganism, a honey bee colony, uses physical cues to sense its size and thus its developmental state.

Comparison of Two New Traps To the Biogents BG-Sentinel Trap For Collecting Aedes albopictus In North Florida.

Multiple surveillance traps are currently available for adult mosquito surveillance. In this study, 2 new types, the Maxttrac™ Uno and Maxttrac™ Breeze mosquito traps, were compared against the BioGents™-Sentinel trap (BGS) for overall success in collecting Aedes albopictus in a 1,027-m2 vehicle enclosure and in the field. The enclosure test results showed both traps collected significantly fewer Ae. albopictus, compared to the BGS trap. The modification of using all BG lures for each trap did not increase the number of mosquitoes collected in the 2 new traps, when compared to collection with the original lures tested in the vehicle enclosure. Field test results showed that BGS trap collected higher numbers of Ae. albopictus than the 2 new traps, regardless of switching lure or not. Overall, the BGS trap was the most effective trap and the 2 new traps could be used as additional tools for the collection of Ae. albopictus.

Effect of Nozzle Orientation on Dispersion of Aqualuer 20-20 Sprayed by a Truck-Mounted Ultra-Low Volume Sprayer Against Caged Aedes aegypti.

Inconsistencies in efficacy of ultra-low volume (ULV) ground applications in the literature are linked to the lack of adjustments in sprayer parameters. To investigate the effect of nozzle orientation of a truck-mounted ULV sprayer on application efficacy, a study was conducted at St. Johns County Fairground, Elkton, FL, during the summer of 2014. The efficacy was assessed by mortality of caged adult Aedes aegypti, spray deposition, and droplet size spectrum up to 122 m from the spray line. Aqualuer 20-20 (20.6% permethrin active ingredient [AI] and 20% piperonyl butoxide [PBO]) was applied at the maximum label rate with the nozzle pointed 45° upward, horizontal (0°), or 30° downward. Mortality was recorded after 24 h, deposition was determined with fluorometry, and droplets were measured with DropVision. Overall, horizontal nozzle angle spraying Aqualuer 20-20 achieved the highest efficacy followed by a 30° downward angle, while a 45° angle showed the least efficacy in open field tests. The mortality data showed complete mortality from a 0° nozzle up to 122 m from the spray line except for 1 location at 122 m in 1 replication. The mortality from a 30° downward orientation was lower beyond 30 m from the spray line, while the mortality from a 45° upward orientation was low close to the spray line and beyond 30 m. Horizontal orientation had higher deposition than other orientations, but the differences were not significant. There was also no significant difference in droplet spectrum from all orientations.

Tensional homeostasis in endothelial cells is a multicellular phenomenon.

Mammalian cells of various types exhibit the remarkable ability to adapt to externally applied mechanical stresses and strains. Because of this adaptation, cells can maintain their endogenous mechanical tension at a preferred (homeostatic) level, which is essential for normal physiological functions of cells and tissues and provides protection against various diseases, including atherosclerosis and cancer. Conventional wisdom is that the cell possesses the ability to maintain tensional homeostasis on its own. Recent findings showed, however, that isolated cells cannot maintain tensional homeostasis. Here we studied the effect of multicellular interactions on tensional homeostasis by measuring traction forces in isolated bovine aortic endothelial cells and in confluent and nonconfluent cell clusters of different sizes. We found that, in isolated cells, the traction field exhibited a highly dynamic and erratic behavior. However, in cell clusters, dynamic fluctuations of the traction field became attenuated with increasing cluster size, at a rate that was faster in nonconfluent than confluent clusters. The driving mechanism of attenuation of traction field fluctuations was statistical averaging of the noise, and the impeding mechanism was nonuniform stress distribution in the clusters, which resulted from intercellular force transmission, known as a "global tug-of-war." These results show that isolated cells could not maintain tensional homeostasis, which confirms previous findings, and that tensional homeostasis is a multicellular phenomenon, which is a novel finding.

Fibronectin Fiber Extension Decreases Cell Spreading and Migration.

The extracellular matrix (ECM) is present in a range of molecular conformations and intermolecular arrangements. Fibronectin (Fn) molecules that constitute fibers within the ECM can exist in a variety of conformations that result from both mechanical stress and chemical factors such as allosteric binding partners. The long-standing hypothesis that conformational changes regulate the binding of cells to Fn fibers has only been tested for mutated molecules of Fn and has yet to be fully evaluated with Fn fibers. Using time-lapse microscopy we examined how mechanical extension of single fibers of Fn affects the adhesion and migration of endothelial cells. Using this single fiber adhesion technique, we show that high levels of mechanical strain applied to Fn fibers decreases the rates of both cell spreading and cell migration. These data indicate a fundamental cellular response to mechanical strain in the ECM that might have important implications for understanding how cells are recruited during tissue development and repair. J. Cell. Physiol. 231: 1728-1736, 2016. © 2015 Wiley Periodicals, Inc.