Phenotype microarray analysis reveals the biotransformation of Fusarium oxysporum f.sp. lycopersici influenced by Bacillus subtilis PBE-8 metabolites.

Herein, Bacillus subtilis PBE-8's biocontrol efficacy was evaluated through physiological and metabolic approaches against Fusarium oxysporum f.sp. lycopersici (FOL). The study elaborates on PBE-8's cell-free filtrate (CFF) antifungal activity through mycelial growth inhibition, metabolite profiling, and substrates utilization patterns. Additionally, under different CFF concentrations, reduction in spore count (94%-55%), biomass (50%), and cytoplasmic bulbous protrusions in mycelia were also observed. Furthermore, the effect of bacterial CFF on FOL metabolism was confirmed through GC-MS. CFF suppresses the concentration of aliphatic amino acids like L-valine, L-leucine, L-Isoleucine, glycine, and fatty acids such as linoleic acid and α- linolenic acid during the co-culturing conditions, which are essential for pathogenicity and resistance against host's systemic acquired resistance. The phenotype microarray assay revealed that CFF-treated FOL shows phenotype loss in 507 (56.58%) out of 896 substrates. Among 507, twenty-seven substrates showed significant phenotype loss, among which four substrates such as L-glutamic acid, L-glutamine, ammonia, and L-arginine are common in different crucial metabolic pathways of FOL, like alanine, aspartate, and glutamate metabolism, arginine and proline, carbon metabolism, arginine biosynthesis, nitrogen metabolism, amino-acyl tRNA synthesis, and biosynthesis of amino acids. The results suggest that PBE-8 CFF has certain antifungal metabolites that hinder the fungal metabolic pathways.

Families Playing Animal Crossing Together: Coping With Video Games During the COVID-19 Pandemic.

The COVID-19 pandemic was stressful for everyone, particularly for families who had to supervise and support children, facilitate remote schooling, and manage work and home life. We consider how families coped with pandemic-related stress using the video game Animal Crossing: New Horizons. Combining a family coping framework with theorizing about media as a coping tool, this interview study of 27 families (33 parents and 37 children) found that parents and children individual coped with pandemic-related stress with media. Parents engaged in protective buffering of their children with media, taking on individual responsibility to cope with a collective problem. Families engaged in communal coping, whereby media helped the family cope with a collective problem, taking on shared ownership and responsibility. We provide evidence for video games as coping tools, but with the novel consideration of family coping with media.

Bacillus subtilis suppresses the charcoal rot disease by inducing defence responses and physiological attributes in soybean.

Endophytes can induce the defence responses and modulates physiological attributes in host plants during pathogen attacks. In the present study, 127 bacterial endophytes (BEs) were isolated from different parts of healthy soybean plant. Among them, two BEs (M-2 and M-4) resulted a significant antagonistic property against Macrophomina phaseolina, causes charcoal rot disease in soybean. The antagonistic potential was evaluated through dual culture plate assay, where M-4 expressed higher antifungal activity than M-2 against M. phaseolina. The M-4 produces cell wall degrading enzymes viz. cellulase (145.71 ± 1.34 µgmL-1), chitinase (0.168 ± 0.0009 unitmL-1) and ß,1-3 endoglucanase (162.14 ± 2.5 µgmL-1), which helps in cell wall disintegration of pathogens. Additionally, M-4 also can produce siderophores, indole-3-acetic acid (IAA) (17.03 ± 1.10 µgmL-1) and had a phosphate solubilization potential (19.89 ± 0.26 µgmL-1). Further, GC-MS profiling of M-4 has been carried out to demonstrate the production of lipophilic secondary metabolites which efficiently suppress the M. phaseolina defensive compounds under co-culture conditions. Bio-efficacy study of M-4 strain shown a significant reduction in disease incidence around 60 and 80% in resistant and susceptible varieties of soybean, respectively. The inoculation of M-4 potentially enhances the physiological attributes and triggers various defence responsive enzymes viz. superoxide dismutase (SOD), phenol peroxidase (PPO), peroxidase (PO) and catalase (CAT). The histopathological study also confirmed that M-4 can reduce the persistence of microsclerotia in root and shoot tissue. Conclusively, M-4 revealed as an efficient biocontrol agent that can uses multifaceted measures for charcoal rot disease management, by suppress the M. phaseolina infection and enhance the physiological attributes of soybean.

Lessons learned from designing an asynchronous remote community approach for behavioral activation intervention for teens.

Adolescent depression is common; however, over 60% of depressed adolescents do not receive mental health care. Digitally-delivered evidence-based psychosocial interventions (EBPIs) may provide an opportunity to improve access and engagement in mental health care. We present a case study that reviews lessons learned from using the Discover - Design - Build - Test (DDBT) model to create, develop, and evaluate a high-fidelity prototype of an app to deliver an EBPI for depression, behavioral activation (BA), on an Asynchronous Remote Communities (ARC) platform (referred to as ActivaTeen). We review work at each stage of the DDBT framework, including initial formative work, iterative design and development work, and an initial feasibility study. We engaged teens with depression, mental health clinicians, and expert evaluators through the process. We found that the DDBT model supported the research team in understanding the requirements for our prototype system, ActivaTeen, and conceiving of and developing specific ideas for implementation. Our work contributes a case study of how the DDBT framework can be applied to adapting an EBPI to a new, scalable and digital format. We provide lessons learned from engaging teens and clinicians with an asynchronous approach to EBPIs and human centered design considerations for teen mental health.

Distribution of vitamin D-binding protein/group-specific component gene subtypes in Kuwaiti population.

BACKGROUND: Vitamin D-binding protein or group-specific component (Gc) is the major plasma carrier protein of Vitamin D. Two single nucleotide polymorphisms, rs7041 (NM_000583.3:c.1296G>T;NP_000574.2:p.Asp432Glu) and rs4588 (c.1307C>A; p.Thr436Lys), in the GC gene result in three major genotypes, that is, GC1F (c.1296T, c.1307C), GC1S (c.1296G, c.1307C), GC2 (c.1296T, c.1307A), and phenotypes such as Gc1F (p.432Asp, p.436Thr), Gc1S (p.432Glu, p.436Thr), and Gc2 (p.432Asp, p.436Lys). Significant variations in the frequencies of GC subtypes (genotypes/phenotypes) are reported in different populations living in different geographical locations, for example, GC1S/Gc1S (c.1296G, c.1307C/p.432Glu, p.436Thr) and GC2/Gc2 (c.1296T, c.1307A/p.432Asp, p.436Lys) are predominant in Caucasians and people living in the northern hemisphere, and GC1F/Gc1F (c.1296T, c.1307C/p.432Asp, p.436Thr) is predominant in Africans. However, frequencies of major GC subtypes are not known in the Kuwaiti population. In this study, we investigated 512 alleles to identify the major GC subtypes in Kuwaiti nationals. METHODS: Genomic DNA was isolated from blood samples of 128 healthy subjects. DNA regions covering the targeted mutations were amplified by PCR. Amplified DNAs were sequenced by the Sanger method and analyzed for specific mutations to determine the GC genotypes and phenotypes. RESULTS: The results identified the presence of four GC genotypes/phenotypes namely GC1F/Gc1F (c.1296T, c.1307C/p.432Asp, p.436Thr), GC1S/Gc1S (c.1296G, c.1307C/p.432Glu, p.436Thr), GC2/Gc2 (c.1296T, c.1307A/p.432Asp, p.436Lys), and GC3/Gc3 (c.1296G;c.1307A/p.432Glu, p.436Lys). Among the allelic subtypes (n = 512), GC1S (c.1296G; c.1307C) (n = 270, 52.7%) was predominant, followed by GC1F (c.1296T; c.1307C) (n = 138, 27%), GC2 (c.1296T; c.1307A) (n = 72, 14%), and GC3 (c.1296G; c.1307A) (n = 32, 6.3%). Three common subtypes, that is, GC1F (c.1296T; c.1307C), GC1S (c.1296G; c.1307C), and GC2 (c.1296T; c.1307A) are well documented in the literature, but GC3 (c.1296T; c.1307A) is an uncommon variant found in our study subjects. CONCLUSION: We found that GC subtype distribution was unique in the Kuwaiti population, with some affinity to Caucasians. Several factors including ancestral origin, migration history, and environmental forces such as solar intensity may be responsible for the unique distribution of GC subtypes in this population.

Phenotypic similarities within the morphologic spectrum of DICER1-associated sarcomas and pleuropulmonary blastoma: Histopathologic features guide diagnosis in the LMIC setting.

Extrapulmonary DICER1-associated sarcomas (DS) can harbor morphological features overlapping with pleuropulmonary blastoma. We report three children with intracranial and genital tract sarcomas, suspected to have DS based on a heterogeneous yet defining combination of spindle-cell sarcomatous and blastemal morphology, with rhabdomyomatous differentiation. Foci of immature cartilage at diagnosis (n = 2/3) and increased neuroepithelial differentiation at recurrence (n = 1) were noted. Morphological suspicion prompted somatic testing at reference centers, confirming likely biallelic, loss-of-function, and "hotspot" missense DICER1 variants in all three tumors. This can serve as a model for this diagnosis in resource-limited settings and has implications for germline testing, surveillance, and tumor management.

Designing Asynchronous Remote Support for Behavioral Activation in Teenagers With Depression: Formative Study.

BACKGROUND: Many teenagers in the United States experience challenges with symptoms of depression, and they lack adequate resources for accessing in-person mental health care. Involving teens and clinicians in designing technologies that use evidence-based practices that reduce barriers to accessing mental health care is crucial. Interventions based on behavioral activation (BA) help teens understand the relationship between mood and activity, help them practice goal-directed behaviors to improve mood, and may be particularly well-suited to delivery via internet-based platforms. OBJECTIVE: This study aims to understand the needs and challenges that teens and mental health clinicians face in depression management and involve them in the design process of a remote intervention that uses asynchronous remote communities. Our goal is to understand the benefits and challenges of adapting BA to an internet-based platform that supports the asynchronous remote community approach as a delivery tool for teen depression management. METHODS: We enrolled mental health clinicians (n=10) and teens (n=8) in separate, private, internet-based groups on Slack (Slack Technologies Inc). They participated in 20-minute design activities for 10 weeks and were then invited to interviews about their experiences in the study. RESULTS: Both teen and clinician participants wanted internet-based support for BA as a supplement to in-person therapy. Although participants perceived the asynchronous format as conducive to supporting accessible care, teens and clinicians raised concerns about safety, privacy, and the moderating of the internet-based group. Design decisions that address these concerns need to be balanced with the potential benefits of learning coping skills, increasing access to mental health care, and promoting asynchronous human connection to support teens. CONCLUSIONS: We discuss considerations for balancing tensions in privacy and safety while designing and selecting internet-based platforms to support remote care and integrating evidence-based support when designing digital technologies for the treatment of teens with depression.

Immunohistochemical screening for mismatch repair protein deficiency in paediatric high-grade gliomas - institutional experience and review of literature.

PURPOSE: Immunohistochemical (IHC) testing for mismatch repair (MMR) deficiency (MMRD) is used as a screening tool to identify microsatellite instability in various cancers (especially colon). This not only identifies hereditary cancer syndromes like Lynch and constitutional mismatch repair deficiency (CMMRD) but also aids in prognostication and prediction of sensitivity to checkpoint inhibitor drugs. There are very few reported studies on MMRD status of pediatric high-grade gliomas (pHGG) and none from the Indian subcontinent. The aim of this study is to evaluate the frequency of MMRD in pHGG and to assess if there is a need for universal screening with immunohistochemistry. METHODS: Paraffin blocks of consecutive cases of pHGG (< 18 years) were retrieved from 2 centres, and IHC with four MMR antibodies - MLH1, PMS2, MSH2 and MSH6 - was performed using tissue microarray-based technique. RESULTS: Three out of nine cases (33%) studied showed loss of staining. One case had loss of MSH2 and MSH6 confirmed by gene sequencing. Eight of the cases were glioblastoma. One case of IDH1-mutated anaplastic astrocytoma showed loss of MLH1 and PMS2 staining. Isolated PMS2 loss was noted in 1 case, where the non-tumour cells also showed loss of staining, indicative CMMRD syndrome. This patient had prior colon cancer with isolated PMS2 loss and responded to check-point inhibitor therapy with nivolumab. CONCLUSION: Our study shows that the frequency of MMRD to be about one-third of pHGG. Universal IHC screening for MMRD in all pHGGs may benefit early diagnosis and play a role in therapeutic decisions. A larger multi-institutional study will help better assess the prevalence and treatment implications in MMRD tumours.

Effects of Oxytocin on Emotion Recognition in Schizophrenia: A Randomized Double-Blind Pilot Study.

BACKGROUND: Schizophrenia (SCZ) is a neurodevelopmental disorder that leads to poor social function. Oxytocin (OXT), a neuropeptide involved in social cognition, is a potential therapeutic agent for alleviating social dysfunction. Therefore, we investigated the effects of intranasal oxytocin (IN-OXT) on emotional processes in experimental interactive social contexts in individuals with SCZ. METHODS: In a male-only parallel randomized placebo-controlled double-blind trial, we investigated the effects of IN-OXT (24 IU) on visual fixation on pictures of faces and emotion recognition in an interactive ball-tossing game that probed processing of social and nonsocial stimuli. RESULTS: Intranasal oxytocin enhanced the recognition of emotions during an emotion-based ball-tossing game. This improvement was specific to the game that included social cue processing. Intranasal oxytocin did not affect eye gaze duration or gaze dwell time on faces in these patients. CONCLUSIONS: An acute low dose of IN-OXT had a modest effect on social cue processing and was limited to emotion recognition. Higher doses and long-term trials targeting emotional processing in SCZ may lead to improved social function.

Langerhans cell histiocytosis in children: A retrospective case series of 126 cases.

BACKGROUND/OBJECTIVES: Langerhans cell histiocytosis (LCH), a rare neoplasm of hematopoietic myeloid precursor cells, is clinically characterized by spontaneously resolving lesions to a progressive life-threatening multisystem disorder. Diagnosing LCH in children is challenging as it mimics other skin disorders. This study describes the varied clinical presentation and disease course in children less than 18 years diagnosed with LCH. METHODS: We performed a retrospective observational study of all cases diagnosed with LCH presenting to a children's hospital in the last 26 years. Data on history, cutaneous and systemic examination, and laboratory evaluation performed, were recorded. RESULTS: A total of 126 children diagnosed with LCH were included in the study. There were 68% cases limited only to skin, and 32% children with multisystem involvement at the initial presentation. Scaly papules were the most common morphologic finding in skin. The skeletal system was the second most common organ system to be affected. Failure to thrive was a common symptom. Progression of skin to systemic involvement was seen in 27.9%. In 76.7%, skin lesions cleared over a period of 2 to 4 years. Complete remission was seen in 56.9% of children over a period of 3 to 7 years, while 8.1% children died of complicationsand 31.8% were lost to follow-up. CONCLUSIONS: Long-term follow-up in this study has shown cutaneous LCH without systemic involvement has a good prognosis. Skin involvement,along with failure to thrive, was the most common clinical presentation in our study. The skeletal system was the second most common organ system involved.

Super protective anti-bacterial coating development with silica-titania nano core-shells.

In the present study, we have developed an anti-bacterial as well as mechanically-strengthened super protective coating material, which can be used as a marine antifouling paint. In this research, silica, titania and silica-titania core-shell nanoparticles were individually prepared via sol-gel and peptization processes. The idea behind the synthesis of core-shell nanoparticles was to utilize the mechanical strength of silica and the antimicrobial property of TiO2 together. These nanoparticles were characterized via dynamic light scattering, UV-Visible spectroscopy, X-ray diffraction, scanning electron microscopy, energy-dispersive X-ray spectroscopy, transmission electron microscopy and X-ray photoelectron spectroscopy. Coating formulations were developed with two types of model binders, i.e., solvent-based polyurethane and water-based poly-acrylic, containing all nanoparticles individually at various concentrations for a better comparative study. These coating formulations were applied onto mild steel for anti-bacterial testing that was performed against Escherichia coli and Bacillus. The nanoparticle concentration was varied from 1% (wt) to 6% (wt). The best anti-bacterial result was obtained with 4% (wt) of silica-titania core-shell nanoparticles prepared via the peptization process among all the nanoparticles. The scratch testing was performed successfully using an Erichsen scratch tester; the formulated PU coating passed up-to 20 N load with good adhesion, impact resistance, flexibility and has shown satisfactory anti-corrosion performance.

Role of silica coated magnetic nanoparticle on cell flocculation, lipid extraction and linoleic acid production from Chlorella pyrenoidosa.

In the present work, it has been observed that magnetic (Fe3O4) - silica core- shell nanoparticles helps in flocculation of Chlorella pyrenoidosa cells with simultaneous production of linoleic acid. The mean particle size in Dynamic light scattering (DLS) of the silica coated magnetic nanoparticle was estimated 444.7 nm. The characterization of nanoparticles was also performed by X-ray diffraction technique (XRD). Apart from flocculation, it has been observed that in presence of magnetic silica core- shell nanoparticles the amount of lipid obtained was four times than that of control. On the contrary, in presence of these nanoparticles, linoleic acid (18:2) has been produced in Chlorella pyrenoidosa cells almost by 80% whereas, it has been noticed only 8.73% in control. This is the first report where the linoleic acid has been obtained as major component of microalgal fatty acid methyl esters (FAME) having important application in nutraceuticals and pharmaceutical sectors.

Relationship of prolonged acoustic startle latency to diagnosis and biotype in the bipolar-schizophrenia network on intermediate phenotypes (B-SNIP) cohort.

BACKGROUND: Latency of the acoustic startle reflex is the time from presentation of the startling stimulus until the response and provides an index of neural processing speed. Latency is prolonged in schizophrenia, is 90% heritable, and predicts conversion to schizophrenia in a high-risk population. The Bipolar-Schizophrenia Network for Intermediate Phenotypes (B-SNIP) consortium investigates neurobiological features found in psychotic disorders spanning diagnostic criteria for schizophrenia (SCZ), schizoaffective disorder (SAD), and psychotic bipolar disorder (BP). We investigated whether differences in startle latency and prepulse inhibition (PPI) occur in probands, their first-degree relatives, and neurobiologically defined subgroups of the probands (Biotypes). METHODS: 1143 subjects were included from the B-SNIP cohort: 143 with SCZ, 178 SCZ relatives (SCZ-Fam), 123 with SAD, 152 SAD relatives (SAD-Fam), 138 BP, 183 BP relatives (BP-Fam), and 226 controls (CON). A Biopac system recorded the eyeblink component of the startle reflex during startle testing. RESULTS: Latency differed by diagnosis (F(3,620) = 5.10, p = 0.002): SCZ, SAD, and BP probands had slower latency than CON, with relatives intermediate. Biotypes 1 and 2 had slower latency than CON (p < 0.031) but Biotype 3 did not differ from CON. PPI did not separate CON from other subjects when analyzed by diagnoses nor when analyzed by biotype. Biotype 1 relatives had slower latency (F(3,663) = 3.49, p = 0.016) and more impaired PPI than Biotype 2 and 3 relatives (F(3,663) = 2.77, p = 0.041). CONCLUSION: Startle latency is prolonged in psychotic disorders that cross traditional diagnostic categories. These data suggest a genetic difference between biotypes that span across clinically defined diagnoses.

Supplementation of Trichoderma improves the alteration of nutrient allocation and transporter genes expression in rice under nutrient deficiencies.

Nutrients are the finite natural resources that are essential for productivity and development of rice and its deficiency causes compromised yield along with reduced immunity against several biotic and abiotic stresses. In this study, the potential of Trichoderma reesei has been investigated as a biofertilizer (BF) to ameliorate nutrient stress in different rice cultivars at physiological, biochemical and molecular levels. The results indicated that cultivar Heena is much more compatible with BF as compared to cultivar Kiran at 50% nutrient limiting condition. Enhancement in physiological attributes and photosynthetic pigments were observed in BF treated Heena seedlings. The localization of biofertilizer in treated roots was further validated by scanning electron micrographs. This result correlated well with the higher levels of Indole acetic acid and Gibberellic acid in biofertilizer treated rice. Similarly, the uptake of micro-nutrients such as Fe, Co, Cu and Mo was found to be 1.4-1.9 fold higher respectively in BF treated Heena seedlings under 50% nutrient deficient condition. Furthermore, different stress ameliorating enzymes Guaiacol peroxidase, Super oxide dismutase, Total Phenolic Content, Phenol Peroxidase, Phenylalanine ammonia lyase and Ascorbate peroxidase in Heena seedlings were also increased by 1.8, 1.4, 1.2, 2.4, 1.2, and 8.3-fold respectively, at 50% nutrient deficient condition. The up-regulation of different micro and macro-nutrients allocation and accumulation; metal tolerance related; auxin synthesis genes in BF treated Heena as compared to 50% nutrient deficient condition was further supported by our findings that the application of biofertilizer efficiently ameliorated the deficiency of nutrients in rice.

Engaging Teenagers in Asynchronous Online Groups to Design for Stress Management.

Teenagers have unique needs for mental wellbeing that can be supported by interactive technologies. Teens also have valuable input in the design of technology, so designers and researchers must seek new methods for involving them in the design process. We enrolled 23 unacquainted teenagers in an Asynchronous Remote Communities (ARC) study consisting of two private online groups. Teens participated in 10 weekly design activities on stress management across three months. We found that teens sought support from technology tailored to their perception of control in stressful contexts, developing sense of self, and varying social needs, including asking for no intervention from others. Teens appreciated that the ARC design experience allowed for flexibility in participation and supported selective disclosure. However, there were limited interactions between teenagers online. Reflecting on our study, we provide design implications for tools to support teenager mental health and discuss how the ARC method can be adapted for designing with teenagers.

Development of coating formulation with silica-titania core-shell nanoparticles against pathogenic fungus.

In the present study, we developed an antifungal coating formulation using silica, titania and silica-titania core-shell nanoparticles individually. The idea behind the synthesis of core-shell nanoparticles was to use the mechanical strength of silica and the antimicrobial property of TiO2 together. These nanoparticles were characterized by dynamic light scattering, transmission electron microscopy, scanning electron microscopy, EDX, FTIR and X-ray diffraction. Silica nanoparticles of 92 nm were prepared by the sol-gel process, while TiO2 nanoparticles and nano-core-shells were prepared through the peptization process with a size of 77 and 144 nm separately. The antifungal effect of the prepared nanoparticles was observed in potato dextrose agar media using the concentration of nanoparticles at 1 wt%. These nanoparticles were incorporated in two types of binder, polyurethane and polyacrylic, with the same concentration of nanoparticles. Coatings were applied on tiles, dried and tested against pathogenic fungus, and fungus growth reduction was observed up to 7-10 days. Coatings developed with TiO2 nanoparticles have shown good growth reduction of pathogenic fungus, but coatings formulated with silica-titania core-shell nanoparticles killed the fungus fusarium completely and have shown around 90% growth reduction for acremonium species also.

Bacterial endophytes modulates the withanolide biosynthetic pathway and physiological performance in Withania somnifera under biotic stress.

Despite the vast exploration of endophytic microbes for growth enhancement in various crops, knowledge about their impact on the production of therapeutically important secondary metabolites is scarce. In the current investigation, chitinolytic bacterial endophytes were isolated from selected medicinal plants and assessed for their mycolytic as well as plant growth promoting potentials. Among them the two most efficient bacterial endophytes namely Bacillus amyloliquefaciens (MPE20) and Pseudomonas fluorescens (MPE115) individually as well as in combination were able to modulate withanolide biosynthetic pathway and tolerance against Alternaria alternata in Withania somnifera. Interestingly, the expression level of withanolide biosynthetic pathway genes (3-hydroxy-3-methylglutaryl co-enzyme A reductase, 1-deoxy-D-xylulose-5-phosphate reductase, farnesyl di-phosphate synthase, squalene synthase, cytochrome p450, sterol desaturase, sterol Δ-7 reductase and sterol glycosyl transferases) were upregulated in plants treated with the microbial consortium under A. alternata stress. In addition, application of microbes not only augmented withaferin A, withanolide A and withanolide B content (1.52-1.96, 3.32-5.96 and 12.49-21.47 fold, respectively) during A. alternata pathogenicity but also strengthened host resistance via improvement in the photochemical efficiency, normalizing the oxidized and non-oxidized fraction, accelerating photochemical and non-photochemical quantum yield, and electron transport rate. Moreover, reduction in the passively dissipated energy of PSI and PSII in microbial combination treated plants corroborate well with the above findings. Altogether, the above finding highlights novel insights into the underlying mechanisms in application of endophytes and emphasizes their capability to accelerate biosynthesis of withanolides in W. somnifera under biotic stress caused by A. alternata.

Endophyte-Mediated Modulation of Defense-Related Genes and Systemic Resistance in Withania somnifera (L.) Dunal under Alternaria alternata Stress.

Endophytes have been explored and found to perform an important role in plant health. However, their effects on the host physiological function and disease management remain elusive. The present study aimed to assess the potential effects of endophytes, singly as well as in combination, in Withania somnifera (L.) Dunal, on various physiological parameters and systemic defense mechanisms against Alternaria alternata Seeds primed with the endophytic bacteria Bacillus amyloliquefaciens and Pseudomonas fluorescens individually and in combination demonstrated an enhanced vigor index and germination rate. Interestingly, plants treated with the two-microbe combination showed the lowest plant mortality rate (28%) under A. alternata stress. Physiological profiling of treated plants showed improved photosynthesis, respiration, transpiration, and stomatal conductance under pathogenic stress. Additionally, these endophytes not only augmented defense enzymes and antioxidant activity in treated plants but also enhanced the expression of salicylic acid- and jasmonic acid-responsive genes in the stressed plants. Reductions in reactive oxygen species (ROS) and reactive nitrogen species (RNS) along with enhanced callose deposition in host plant leaves corroborated well with the above findings. Altogether, the study provides novel insights into the underlying mechanisms behind the tripartite interaction of endophyte-A. alternata-W. somnifera and underscores their ability to boost plant health under pathogen stress.IMPORTANCEW. somnifera is well known for producing several medicinally important secondary metabolites. These secondary metabolites are required by various pharmaceutical sectors to produce life-saving drugs. However, the cultivation of W. somnifera faces severe challenge from leaf spot disease caused by A. alternata To keep pace with the rising demand for this plant and considering its capacity for cultivation under field conditions, the present study was undertaken to develop approaches to enhance production of W. somnifera through intervention using endophytes. Application of bacterial endophytes not only suppresses the pathogenicity of A. alternata but also mitigates excessive ROS/RNS generation via enhanced physiological processes and antioxidant machinery. Expression profiling of plant defense-related genes further validates the efficacy of bacterial endophytes against leaf spot disease.

Understanding pivotal experiences in behavior change for the design of technologies for personal wellbeing.

Most health technologies are designed to support people who have already decided to work toward better health. Thus, there remains an opportunity to design technologies to help motivate people who have not yet decided to make a change. Understanding the experiences of people who have already started to make a health behavior change and how they made a pivotal decision can be useful in understanding how to design such tools. In this paper, we describe results from data collected in 2 phases. Phase 1 consisted of 127 surveys and 13 interviews with adults who have already accomplished behavior change(s). Phase 2 consisted of 117 surveys and 12 interviews with adults who have either already accomplished their behavior change(s) or are currently working toward them. We identified four factors that lead to pivotal experiences: (1) prolonged discontent and desire to change, (2) significant changes that increase fear or hope of future, (3) increased understanding of one's behavior and personal data, and (4) social accountability. We also describe a design space for designing technology-based interventions for encouraging people to decide to make a change to improve their health. Based on feedback from participants, we discuss opportunities for further exploration of the design space for people who are not yet motivated to change and for ethical considerations for this type of intervention.

Lessons from Practice: Designing Tools to Facilitate Individualized Support for Quitting Smoking.

Many health care providers, with a variety of trainings, counsel clients on quitting smoking on a day-to-day basis. In their clinical practice, they draw from and adapt guidelines and research-based strategies to fit individual client situations and challenges. Designers of technologies to support quitting smoking can learn from these real world practices to create tools that better adapt to individual differences. We present findings from interviews with 28 providers with diverse experiences in smoking cessation counselling. Through analysis of their individualization strategies, challenges, and perceptions of technology, we find that providers: (1) individualize context appropriate coping strategies by involving clients in brainstorming, (2) emphasize the need to support nicotine withdrawal in clients, (3) mitigate social triggers and mediate social support for clients, and (4) need to navigate dependencies with other providers for managing medications and comorbid health conditions of clients. With this empirical understanding, we extend the discussion on the design of technology to support quitting smoking, highlight current barriers to individualization, and suggest future opportunities to address these barriers.