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In this work, gene expression in autism spectrum disorder (ASD) is analyzed with the goal of selecting the most attributed genes and performing classification. The objective was achieved by utilizing a combination of various statistical filters and a wrapper-based geometric binary particle swarm optimization-support vector machine (GBPSO-SVM) algorithm. The utilization of different filters was accentuated by incorporating a mean and median ratio criterion to remove very similar genes. The results showed that the most discriminative genes that were identified in the first and last selection steps included the presence of a repetitive gene (CAPS2), which was assigned as the gene most highly related to ASD risk. The merged gene subset that was selected by the GBPSO-SVM algorithm was able to enhance the classification accuracy.

In this research work, numerical simulations are performed to correlate the photovoltaic parameters with various internal and external factors influencing the performance of solar cells. Single-diode modeling approach is utilized for this purpose and theoretical investigations are compared with the reported experimental evidences for organic and inorganic solar cells at various electrical and thermal conditions. Electrical parameters include parasitic resistances (Rs and Rp) and ideality factor (n), while thermal parameters can be defined by the cells temperature (T). A comprehensive analysis concerning broad spectral variations in the short circuit current (Isc), open circuit voltage (Voc), fill factor (FF) and efficiency (η) is presented and discussed. It was generally concluded that there exists a good agreement between the simulated results and experimental findings. Nevertheless, the controversial consequence of temperature impact on the performance of organic solar cells necessitates the development of a complementary model which is capable of well simulating the temperature impact on these devices performance.

Cryptococcus gattii is a fungal pathogen that can cause life-threatening respiratory and disseminated infections in immune-competent and immune-suppressed individuals. Currently, there are no standardized vaccines against cryptococcosis in humans, underlying an urgent need for effective therapies and/or vaccines. In this study, we evaluated the efficacy of intranasal immunization with C. gattii cell wall associated (CW) and/or cytoplasmic (CP) protein preparations to induce protection against experimental pulmonary C. gattii infection in mice. BALB/c mice immunized with C. gattii CW and/or CP protein preparations exhibited a significant reduction in pulmonary fungal burden and prolonged survival following pulmonary challenge with C. gattii. Protection was associated with significantly increased pro-inflammatory and Th1-type cytokine recall responses, in vitro and increased C. gattii-specific antibody production in immunized mice challenged with C. gattii. A number of immunodominant proteins were identified following immunoblot analysis of C. gattii CW and CP protein preparations using sera from immunized mice. Immunization with a combined CW and CP protein preparation resulted in an early increase in pulmonary T cell infiltrates following challenge with C. gattii. Overall, our studies show that C. gattii CW and CP protein preparations contain antigens that may be included in a subunit vaccine to induce prolonged protection against pulmonary C. gattii infection.

Fuzzy graphs are very important when we are trying to understand and study complex systems with uncertain and not exact information. Among different types of fuzzy graphs, cubic fuzzy graphs are special due to their ability to represent the membership degree of both vertices and edges using intervals and fuzzy numbers, respectively. To figure out how things are connected in cubic fuzzy graphs, we need to know about cubic α-strong, cubic β-strong and cubic δ-weak edges. These concepts better help in making decisions, solving problems and analyzing things like transportation, social networks and communication systems. The applicability of connectivity and comprehension of cubic fuzzy graphs have urged us to discuss connectivity in the domain of cubic fuzzy graphs. In this paper, the terms partial cubic α-strong and partial cubic δ-weak edges are introduced for cubic fuzzy graphs. The bounds and exact expression of connectivity index for several cubic fuzzy graphs are estimated. The average connectivity index for cubic fuzzy graphs is also defined and some results pertaining to these concepts are proved in this paper. The results demonstrate that removing some vertices or edges may cause a change in the value of connectivity index or average connectivity index, but the change will not necessarily be related to both values. This paper also defines the concepts of partial cubic connectivity enhancing node and partial cubic connectivity reducing node and some related results are proved. Furthermore, the concepts of cubic α-strong, cubic β- strong, cubic δ-weak edge, partial cubic α-strong and partial cubic δ-weak edges are utilized to identify areas most affected by a tsunami resulting from an earthquake. Finally, the research findings are compared with the existing methods to demonstrate their suitability and creativity., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Shi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Degenerative Cervical Myelopathy (DCM) is one of the most common degenerative disorders of cervical spine and sources of cord dysfunction in adults. It usually manifests with neurologic presentations such as loss of dexterity and gait issues. Treatment for moderate and severe cases of DCM is surgical decompression of the region. There are many approaches available for surgical intervention which could be categorized into anterior and posterior based on the side of neck where operation takes place. Additionally, for certain cases the hybridized anterior-posterior combined surgery is indicated. While there are many technical differences between these approaches with each having its own advantages, the complications and safety profiles of them are not fully disclosed. This protocol aims to systematically search for current reports on complications of surgical decompression methods of DCM and pool them for robust evidence generation., Method: Search will be carried out in PubMed, Scopus, and Cochrane databases for retrospective and prospective surgical series, cohorts, or trials being performed for DCM with at least a sample size of 20 patients. Query strings will be designed to capture reports with details of complications with no year limit. Studies not being original (e.g., review articles, case reports, etc.), not in English, having patients younger than 18-years-old, and not reporting at least one complication will be excluded. Two independent reviewers will review the titles and abstracts for first round of screening. Full text of retrieved studies from previous round will be screened again by the same reviewers. In case of discrepancy, the third senior reviewer will be consulted. Eligible studies will then be examined for data extraction where data will be recorded into standardized form. Cumulative incidence and 95% confidence intervals of complication will be then pooled based on generalized linear mixed models with consideration of approach of surgery as subgroups. Heterogeneity tests will be performed for assessment of risk of bias., Discussion: This systematic review is aimed at providing practical information for spine surgeons on the rates of complications of different surgical approaches of DCM decompression. Proper decision-making regarding the surgical approach in addition to informing patients could be facilitated through results of this investigation., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Mohammadi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

In human medical practice, a hematological rule of three has been validated for healthy human populations. One such formula is estimating hemoglobin (Hb) levels as 1/3rd of Packed Cell Volume (PCV). However, no such hematological formulae have been devised and validated for veterinary medical practice. The present study was devised with an aim to evaluate the relationship between hemoglobin (Hb) concentration and Packed Cell Volume (PCV) in camels (n = 215) being reared under pastoralism, and to devise a simple pen-side hematological formula for estimation of Hb from PCV. The PCV was determined through microhematocrit method whereas Hb estimation by cyanmethaemoglobin method (HbD). The Hb was also calculated as 1/3rd of PCV and was dubbed as calculated Hb (HbC). Overall HbD and HbC were significantly (P≥0.05) different. Similar results were attained for all study groups i.e. males (n = 94) and females (n = 121), and young (n = 85) and adult (n = 130) camels. The corrected Hb (CHb) was deduced through regression prediction equation attained from linear regression model. Scatterplots were drawn, linear regression was carried out, and Bland Altman chart was built for agreement of both methods of Hb estimation. A non-significant (P≥0.05) difference was noticed between HbD and CHb. Bland Altman agreement analysis revealed satisfactory agreement between HbD and CHb and the data was distributed closely around the mean difference line (Mean = 0.1436, 95% CI = 3.00, -2.72). A simplified pen-side hematological formula for deducing Hb concentration from PCV is accordingly recommended viz. Hb concentration (g/dL) = 0.18(PCV)+5.4 for all age and gender groups of camels instead of its calculation as one-third of PCV., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests it will helpful to manage the health of animals, and also helpful for professionals This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2023 Farooq et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: To assess access to general and disability-related health care among people with disabilities in the Maldives., Methods: This study uses data from a case-control study (n = 711) nested within a population-based, nationally representative survey to compare health status and access to general healthcare amongst people with and without disabilities. Cases and controls were matched by gender, location and age. Unmet need for disability-related healthcare is also assessed. Multivariate regression was used for comparisons between people with and without disabilities., Results: People with disabilities had poorer levels of health compared to people without disabilities, including poorer self-rated health, increased likelihood of having a chronic condition and of having had a serious health event in the previous 12 months. Although most people with and without disabilities sought care when needed, people with disabilities were much more likely to report difficulties when routinely accessing healthcare services compared to people without disabilities. Additionally, 24% of people with disabilities reported an unmet need for disability-related healthcare, which was highest amongst people with hearing, communication and cognitive difficulties, as well as amongst older adults and people living in the lowest income per capita quartile. Median healthcare spending in the past month was modest for people with and without disabilities. However, people with disabilities appear to have high episodic healthcare costs, such as for disability-related healthcare and when experiencing a serious health event., Conclusions: This study found evidence that people with disabilities experience unmet needs for both disability-related and general healthcare. There is therefore evidence that people with disabilities in the Maldives are falling behind in core components relevant to UHC: availability of all services needed, and quality and affordability of healthcare., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 Banks et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

With a limited arsenal of available antifungal drugs and drug-resistance emergence, strategies that seek to reduce Candida immune evasion and virulence could be a promising alternative option. Harnessing metal homeostasis against C. albicans has gained wide prominence nowadays as a feasible antifungal strategy. Herein, the effect of magnesium (Mg) deprivation on the immune evasion mechanisms of C. albicans is demonstrated. We studied host pathogen interaction by using the THP-1 cell line model and explored the avenue that macrophage-mediated killing was enhanced under Mg deprivation, leading to altered cytokine (TNFα, IL-6 and IL10) production and reduced pyroptosis. Insights into the mechanisms revealed that hyphal damage inside the macrophage was diminished under Mg deprivation. Additionally, Mg deprivation led to cell wall remodelling; leading to enhanced β-1,3-glucan exposure, crucial for immune recognition, along with concomitant alterations in chitin and mannan levels. Furthermore, vacuole homeostasis was disrupted under Mg deprivation, as revealed by abrogated morphology and defective acidification of the vacuole lumen. Together, we demonstrated that Mg deprivation affected immune evasion mechanisms by: reduced hyphal damage, enhanced β-1,3-glucan exposure and altered vacuole functioning. The study establishes that Mg availability is indispensable for successful C. albicans immune evasion and specific Mg dependent pathways could be targeted for therapy., Competing Interests: The authors have declared that no competing interests exist.

Climate change is expected to impact a large number of organisms in many ecosystems, including several threatened mammals. A better understanding of climate impacts on species can make conservation efforts more effective. The Himalayan ibex (Capra ibex sibirica) and blue sheep (Pseudois nayaur) are economically important wild ungulates in northern Pakistan because they are sought-after hunting trophies. However, both species are threatened due to several human-induced factors, and these factors are expected to aggravate under changing climate in the High Himalayas. In this study, we investigated populations of ibex and blue sheep in the Pamir-Karakoram mountains in order to (i) update and validate their geographical distributions through empirical data; (ii) understand range shifts under climate change scenarios; and (iii) predict future habitats to aid long-term conservation planning. Presence records of target species were collected through camera trapping and sightings in the field. We constructed Maximum Entropy (MaxEnt) model on presence record and six key climatic variables to predict the current and future distributions of ibex and blue sheep. Two representative concentration pathways (4.5 and 8.5) and two-time projections (2050 and 2070) were used for future range predictions. Our results indicated that ca. 37% and 9% of the total study area (Gilgit-Baltistan) was suitable under current climatic conditions for Himalayan ibex and blue sheep, respectively. Annual mean precipitation was a key determinant of suitable habitat for both ungulate species. Under changing climate scenarios, both species will lose a significant part of their habitats, particularly in the Himalayan and Hindu Kush ranges. The Pamir-Karakoram ranges will serve as climate refugia for both species. This area shall remain focus of future conservation efforts to protect Pakistan's mountain ungulates., Competing Interests: The authors have declared that no competing interests exist.

Objectives: To estimate the prevalence of disability among Syrian refugees living in Sultanbeyli district, Istanbul and compare people with and without disabilities in terms of demographic and socio-economic characteristics., Methods: Using the municipality refugee database as the sampling frame, 80 clusters of 50 people (aged 2+ years) were selected using probability proportionate to size sampling of clusters and random selection of households within clusters. Disability assessment included: i) self-reported difficulties in functioning (using the Washington Group Short Set-Enhanced tool and Child Functioning Modules), ii) Rapid Assessment of Musculoskeletal Impairment and iii) screening for symptoms of common mental disorders for children aged 8-17., Results: The overall prevalence of disability was 24.7% (95% CI 22.1-27.4), when including people self-reporting a lot of difficulty/cannot do in at least functional domain (15%, 95% CI 13.1-17.2), moderate/severe MSI (8.7%, 95% CI 7.6-9.9), and/or symptomatic anxiety, depression and PTSD among children 8-17 (21.0%, 95% CI 18.2-23.9). Men with disabilities were significantly less likely to be in paid work compared to their peers without disabilities (aOR 0.3 95% CI 0.2-0.5). Overall 60% of households included at least one person with a disability. Households with at least one person with a disability had a significantly higher dependency ratio, lower proportion of working-age adults in paid work, and were more likely to be female headed and in receipt of social protection schemes (p<0.05)., Conclusion: Disability is common among Syrian refugees in Sultanbeyli. People with disabilities in this setting experience greater vulnerability to poverty and exclusion from work, highlighting an urgent need for inclusive services, programmes and policies that are developed and implemented in partnership with people with disabilities., Competing Interests: The authors have declared that no competing interests exist.

Pakistan's total estimated snow leopard habitat is about 80,000 km2 of which about half is considered prime habitat. However, this preliminary demarcation was not always in close agreement with the actual distribution-the discrepancy may be huge at the local and regional level. Recent technological developments like camera trapping and molecular genetics allow for collecting reliable presence records that could be used to construct realistic species distribution based on empirical data and advanced mathematical approaches like MaxEnt. The current study followed this approach to construct an accurate distribution of the species in Pakistan. Moreover, movement corridors, among different landscapes, were also identified through circuit theory. The probability of habitat suitability, generated from 98 presence points and 11 environmental variables, scored the snow leopard's assumed range in Pakistan, from 0 to 0.97. A large portion of the known range represented low-quality habitat, including areas in lower Chitral, Swat, Astore, and Kashmir. Conversely, Khunjerab, Misgar, Chapursan, Qurumber, Broghil, and Central Karakoram represented high-quality habitats. Variables with higher contributions in the MaxEnt model were precipitation during the driest month (34%), annual mean temperature (19.5%), mean diurnal range of temperature (9.8%), annual precipitation (9.4%), and river density (9.2). The model was validated through receiver operating characteristic (ROC) plots and defined thresholds. The average test AUC in Maxent for the replicate runs was 0.933 while the value of AUC by ROC curve calculated at 0.15 threshold was 1.00. These validation tests suggested a good model fit and strong predictive power. The connectivity analysis revealed that the population in the Hindukush landscape appears to be more connected with the population in Afghanistan as compared to other populations in Pakistan. Similarly, the Pamir-Karakoram population is better connected with China and Tajikistan, while the Himalayan population was connected with the population in India. Based on our findings we propose three model landscapes to be considered under the Global Snow Leopard Ecosystem Protection Program (GSLEP) agenda as regional priority areas, to safeguard the future of the snow leopard in Pakistan and the region. These landscapes fall within mountain ranges of the Himalaya, Hindu Kush and Karakoram-Pamir, respectively. We also identified gaps in the existing protected areas network and suggest new protected areas in Chitral and Gilgit-Baltistan to protect critical habitats of snow leopard in Pakistan., Competing Interests: The authors have declared that no competing interests exist.

Eclipta alba L., also known as false daisy, is well known and commercially attractive plant with excellent hepatotoxic and antidiabetic activities. Light is considered a key modulator in plant morphogenesis and survival by regulating important physiological cascades. Current study was carried out to investigate growth and developmental aspects of E. alba under differential effect of multispectral lights. In vitro derived callus culture of E. alba was exposed to multispectral monochromatic lights under controlled aseptic conditions. Maximum dry weight was recorded in culture grown under red light (11.2 g/L) whereas negative effect was observed under exposure of yellow light on callus growth (4.87 g/L). Furthermore, red light significantly enhanced phenolics and flavonoids content (TPC: 57.8 mg/g, TFC: 11.1 mg/g) in callus cultures compared to rest of lights. HPLC analysis further confirmed highest accumulation of four major compounds i.e. coumarin (1.26 mg/g), eclalbatin (5.00 mg/g), wedelolactone (32.54 mg/g) and demethylwedelolactone (23.67 mg/g) and two minor compounds (β-amyrin: 0.38 mg/g, luteolin: 0.39 mg/g) in red light treated culture whereas stigmasterol was found optimum (0.22 mg/g) under blue light. In vitro based biological activities including antioxidant, antidiabetic and lipase inhibitory assays showed optimum values in cultures exposed to red light, suggesting crucial role of these phytochemicals in the enhancement of the therapeutic potential of E. alba. These results clearly revealed that the use of multispectral lights in in vitro cultures could be an effective strategy for enhanced production of phytochemicals., Competing Interests: The authors have declared that no competing interests exist.

Background: One quarter of the global population is of menstruating age, yet menstruation is shrouded in discrimination and taboos. Disability also carries stigma, so disabled people may face layers of discrimination when they are menstruating. The objective of the review is to assess the menstrual hygiene requirements of disabled people, the barriers they face, and the available interventions to help them manage their menstruation hygienically and with dignity., Methods: Eligible studies, gathered across all countries, were identified by conducting searches across four databases (MEDLINE, PubMed, EMBASE, Global Health) in May 2017, with alerts set on each database to highlight new titles added until April 2018. Eligible studies incorporated analyses relevant to menstruating disabled people and/or how their carers provide support during their menstrual cycle., Results: The 22 studies included were published since 1976; the majority after 2010 (n = 12; 55%). One study was a quasi-experiment; all others were observational. Most studies (n = 15; 68%) were from high income countries and most (n = 17; 77%) focused on people with intellectual impairments, so the review findings focus on this group and their carers. Outcomes investigated include choice and preference of menstrual product, ability to manage menstrual hygiene and coping strategies applied. Barriers faced included a lack of standardised guidance for professional carers; a lack of menstruation training, information and support provided to people with intellectual impairments and their carers; a lack of understanding of severity of symptoms experienced by people with intellectual impairments, the high cost of menstrual products and lack of appropriate options for people with physical impairments. Few interventions were found, and strategies for menstrual hygiene management applied by carers of persons with intellectual impairments include limiting the disabled person's movements when menstruating and suppressing their menstruation., Conclusions: Little evidence was identified on the requirements of disabled people and their carers in managing their menstruation, and only one intervention, but a range of barriers were identified. This gap in evidence is important, as the consequences of failing to meet menstrual hygiene needs of disabled people includes shame, social isolation, and even sterilisation., Systematic Review Registration: PROSPERO CRD42018095497., Competing Interests: The authors have declared that no competing interests exist.

Among the several mechanisms of multidrug resistance (MDR), overexpression of drug efflux pumps CaCdr1p and CaMdr1p belonging to ATP binding cassette (ABC) and major facilitator superfamily (MFS) respectively remain the predominant mechanisms of candidal infections. Therefore inhibiting or modulating the function of these transporters continues to draw attention as effective strategy to combat MDR. We have previously reported the antifungal potential of Geraniol (Ger), a natural monoterpenoid from Palmarosa oil, against Candida albicans. Herein, we explored the fungicidal nature of Ger. The Rhodamine 6G (R6G) and Nile red accumulation confirms the specific effect on CaCdr1p. Mechanistic insights with Candida cells overexpressing CaCdr1p and CaMdr1p revealed that Ger specifically modulates CaCdr1p activity. Kinetic studies further unraveled the competitive inhibition of Ger for R6G efflux as evident from increased apparent Km without affecting Vmax value. The effect of Ger on CaCdr1p was substantiated by molecular docking analyses, which depicted in-silico binding affinity of Ger with CaCdr1p and explored that Ger binds to the active site of CaCdr1p with higher binding energy. Although RT-PCR and western blot revealed no change in expressions of CDR1 and CaCdr1p, confocal microscopy images however depicted CaCdr1p mislocalization in presence of Ger. Interestingly, Ger was synergistic (FICI<0.5) with fluconazole (FLC) which is a well known antifungal drug. Furthermore, Ger sensitizes the FLC sensitive and resistant clinical matched pair of isolates Gu4/Gu5 and led to abrogated R6G efflux and depleted ergosterol. Furthermore, Rhodamine B labeling demonstrates altered mitochondrial potential with Ger which suggest possible linkage of dysfunctional mitochondria with CaCdr1p activity. We also estimated phenotypic virulence marker extracellular phospholipase activity which was considerably diminished along with inhibited cell adherence and biofilm biomass. Lastly, antifungal efficacy of Ger was demonstrated by enhanced survival of Caenorhabditis elegans model and negligible hemolytic activity (20%). Together, modulation of efflux pump activity by Ger and FLC synergism represent a promising approach for combinatorial treatment of candidiasis., Competing Interests: The authors have declared that no competing interests exist.

Habitat suitability models are useful to understand species distribution and to guide management and conservation strategies. The grey wolf (Canis lupus) has been extirpated from most of its historic range in Pakistan primarily due to its impact on livestock and livelihoods. We used non-invasive survey data from camera traps and genetic sampling to develop a habitat suitability model for C. lupus in northern Pakistan and to explore the extent of connectivity among populations. We detected suitable habitat of grey wolf using a maximum entropy approach (Maxent ver. 3.4.0) and identified suitable movement corridors using the Circuitscape 4.0 tool. Our model showed high levels of predictive performances, as seen from the values of area under curve (0.971±0.002) and true skill statistics (0.886±0.021). The main predictors for habitat suitability for C. lupus were distances to road, mean temperature of the wettest quarter and distance to river. The model predicted ca. 23,129 km2 of suitable areas for wolf in Pakistan, with much of suitable habitat in remote and inaccessible areas that appeared to be well connected through vulnerable movement corridors. These movement corridors suggest that potentially the wolf range can expand in Pakistan's Northern Areas. However, managing protected areas with stringent restrictions is challenging in northern Pakistan, in part due to heavy dependence of people on natural resources. The habitat suitability map provided by this study can inform future management strategies by helping authorities to identify key conservation areas.

Molecular screening of new patients suspected for TB could help in the effective control of TB in Pakistan as it is a high TB burden country. It will be informative to understand the prevalence of multi drug resistance for a better drug regimen management in this geographical area. The Rifampicin resistance determining region (RRDR) sequencing was used to identify mutations associated with drug resistance in DNA extracts from 130 known multidrug resistant (MDR) cultured strains and compared with mutations observed in DNA extracts directly from 86 sputum samples from consecutive newly diagnosed cases in Lahore, Pakistan. These newly diagnosed samples were positive for smear microscopy, chest X-ray and presumed sensitive to first line drugs. In the known MDR group the most frequent mutations conferring resistance were found in rpoB531 (n = 51, 39.2%). In the newly diagnosed tuberculosis group with no history of MDR, mutations in rpoB531 were seen in 10 of the samples (11.6%). Collectively, all mutations in the RRDR region studied were observed in 80 (61.5%) of known MDR cases and in 14 (16.3%) of the newly diagnosed cases. Using the RRDR as a surrogate marker for MDR, sequences for the newly diagnosed (presumed sensitive) group indicate much higher levels of MDR than the 3.9% WHO 2015 global estimate and suggests that molecular screening directly from sputum is urgently required to effectively address the detection and treatment gaps to combat MDR in this high burden country.

This study explored the antifungal potential of perillyl alcohol (PA), a natural monoterpene alcohol, against most prevalent human fungal pathogen C. albicans, its clinical isolates and four non-albicans species of Candida. To resolve the potential mechanisms, we used whole genome transcriptome analyses of PA treated Candida cells to examine the affected cellular circuitry of this pathogen. The transcriptome data revealed a link between calcineurin signaling and PA as among the several categories of PA responsive genes the down regulation of calcineurin signaling gene CNB1 was noteworthy which was also confirmed by both molecular docking and susceptibility assays. We observed that PA treated Candida phenocopied compromised calcineurin pathway stress responses and turned sensitive to alkaline pH, ionic, membrane, salinity, endoplasmic reticulum and serum stresses. Indispensability of functional calcineurin was further confirmed as calcineurin mutant was hypersensitive to PA while constitutively expressed calcineurin strain remained resistant. We explored that PA leads to perturbed membrane integrity as depicted through depleted ergosterol levels and disrupted pH homeostasis. Moreover, PA caused cell wall damage which was evident from hypersensitivity against cell wall perturbing agents (congo red, calcoflour white), SEM and enhanced rate of cell sedimentation. Furthermore, PA inhibited potential virulence traits including morphological transition, biofilm formation and displayed diminished capacity to adhere both to the polystyrene surface and buccal epithelial cells. The study also revealed that PA leads to cell cycle arrest and mitochondrial dysfunction in C. albicans. Together, the present study provides enough evidence for further work on PA so that better strategies could be employed to treat Candida infections., Competing Interests: The authors have declared that no competing interests exist.

A field study was conducted to compare the formationand bacterial communities of rhizosheaths of wheat grown under wheat-cotton and wheat-rice rotation and to study the effects of bacterial inoculation on plant growth. Inoculation of Azospirillum sp. WS-1 and Bacillus sp. T-34 to wheat plants increased root length, root and shoot dry weight and dry weight of rhizosheathsoil when compared to non-inoculated control plants, and under both crop rotations. Comparing both crop rotations, root length, root and shoot dry weight and dry weight of soil attached with roots were higher under wheat-cotton rotation. Organic acids (citric acid, malic acid, acetic acid and oxalic acid) were detected in rhizosheaths from both rotations, with malic acid being most abundant with 24.8±2 and 21.3±1.5 μg g(-1) dry soil in wheat-cotton and wheat-rice rotation, respectively. Two sugars (sucrose, glucose) were detected in wheat rhizosheath under both rotations, with highest concentrations of sucrose (4.08±0.5 μg g(-1) and 7.36±1.0 μg g(-1)) and glucose (3.12±0.5 μg g(-1) and 3.01± μg g(-1)) being detected in rhizosheaths of non-inoculated control plants under both rotations. Diversity of rhizosheath-associated bacteria was evaluated by cultivation, as well as by 454-pyrosequencing of PCR-tagged 16S rRNA gene amplicons. A total of 14 and 12 bacterial isolates predominantly belonging to the genera Arthrobacter, Azospirillum, Bacillus, Enterobacter and Pseudomonaswere obtained from the rhizosheath of wheat grown under wheat-cotton and wheat-rice rotation, respectively. Analysis of pyrosequencing data revealed Proteobacteria, Bacteriodetes and Verrucomicrobia as the most abundant phyla in wheat-rice rotation, whereas Actinobacteria, Firmicutes, Chloroflexi, Acidobacteria, Planctomycetes and Cyanobacteria were predominant in wheat-cotton rotation. From a total of 46,971 sequences, 10.9% showed ≥97% similarity with 16S rRNA genes of 32 genera previously shown to include isolates with plant growth promoting activity (nitrogen fixation, phosphate-solubilization, IAA production). Among these, the most predominant genera were Arthrobacter, Azoarcus, Azospirillum, Bacillus, Cyanobacterium, Paenibacillus, Pseudomonas and Rhizobium.

We previously demonstrated that iron deprivation enhances drug susceptibility of Candida albicans by increasing membrane fluidity which correlated with the lower expression of ERG11 transcript and ergosterol levels. The iron restriction dependent membrane perturbations led to an increase in passive diffusion and drug susceptibility. The mechanisms underlying iron homeostasis and multidrug resistance (MDR), however, are not yet resolved. To evaluate the potential mechanisms, we used whole genome transcriptome and electrospray ionization tandem mass spectrometry (ESI-MS/MS) based lipidome analyses of iron deprived Candida cells to examine the new cellular circuitry of the MDR of this pathogen. Our transcriptome data revealed a link between calcineurin signaling and iron homeostasis. Among the several categories of iron deprivation responsive genes, the down regulation of calcineurin signaling genes including HSP90, CMP1 and CRZ1 was noteworthy. Interestingly, iron deprived Candida cells as well as iron acquisition defective mutants phenocopied molecular chaperone HSP90 and calcineurin mutants and thus were sensitive to alkaline pH, salinity and membrane perturbations. In contrast, sensitivity to above stresses did not change in iron deprived DSY2146 strain with a hyperactive allele of calcineurin. Although, iron deprivation phenocopied compromised HSP90 and calcineurin, it was independent of protein kinase C signaling cascade. Notably, the phenotypes associated with iron deprivation in genetically impaired calcineurin and HSP90 could be reversed with iron supplementation. The observed down regulation of ergosterol (ERG1, ERG2, ERG11 and ERG25) and sphingolipid biosynthesis (AUR1 and SCS7) genes followed by lipidome analysis confirmed that iron deprivation not only disrupted ergosterol biosynthesis, but it also affected sphingolipid homeostasis in Candida cells. These lipid compositional changes suggested extensive remodeling of the membranes in iron deprived Candida cells. Taken together, our data provide the first novel insight into the intricate relationship between cellular iron, calcineurin signaling, membrane lipid homeostasis and drug susceptibility of Candida cells.