Chromatin dynamics plays an important role in medical mycology. Here, we present a protocol to identify nucleosome occupancy and positioning in the fungal pathogen Candida glabrata (Nakaseomyces glabratus). This process involves micrococcal nuclease (MNase) digestion of the chromatin of C. glabrata cells (grown in RPMI medium or inside macrophages), followed by the purification of mononucleosomal DNA fragments. We describe steps for standardizing MNase digestion and identifying genome-wide dynamic nucleosomes using a next-generation sequencing approach. For complete details on the use and execution of this protocol, please refer to Kumar et al.1.

Abstract Background Vector control based on indoor residual spraying (IRS) is one of the main components of the visceral leishmaniasis (VL) elimination programme in India. Dichlorodiphenyltrichloroethane (DDT) was used for IRS until 2015 and was later replaced by the synthetic pyrethroid alpha-cypermethrin. Both classes of insecticides share the same target site, the voltage-gated sodium channel (Vgsc). As high levels of resistance to DDT have been documented in the local sand fly vector, Phlebotomus argentipes, it is possible that mutations in the Vgsc gene could provide resistance to alpha-cypermethrin, affecting current IRS pyrethroid-based vector control. Methods This study aimed to compare frequencies of knockdown resistance (kdr) mutations in Vgsc between two sprayed and two unsprayed villages in Bihar state, India, which had the highest VL burden of the four endemic states. Across four villages, 350 female P. argentipes were collected as part of a 2019 molecular xenomonitoring study. DNA was extracted and used for sequence analysis of the IIS6 fragment of the Vgsc gene to assess the presence of kdr mutations. Results Mutations were identified at various positions, most frequently at codon 1014, a common site known to be associated with insecticide resistance in mosquitoes and sand flies. Significant inter-village variation was observed, with sand flies from Dharampur, an unsprayed village, showing a significantly higher proportion of wild-type alleles (55.8%) compared with the three other villages (8.5–14.3%). The allele differences observed across the four villages may result from selection pressure caused by previous exposure to DDT. Conclusions While DDT resistance has been reported in Bihar, P. argentipes is still susceptible to pyrethroids. However, the presence of kdr mutations in sand flies could present a threat to IRS used for VL control in endemic villages in India. Continuous surveillance of vector bionomics and insecticide resistance, using bioassays and target genotyping, is required to inform India’s vector control strategies and to ensure the VL elimination target is reached and sustained. Graphical Abstract

Immune evasion is critical for fungal virulence. However, how the human opportunistic pathogen Candida glabrata (Cg) accomplishes this is unknown. Here, we present the first genome-wide nucleosome map of the macrophage-internalized Cg consisting of ∼12,000 dynamic and 70,000 total nucleosomes. We demonstrate that CgSnf2 (SWI/SNF chromatin remodeling complex-ATPase subunit)-mediated chromatin reorganization in macrophage-internalized Cg upregulates and downregulates the immunosuppressive seven-gene mannosyltransferase-cluster (CgMT-C) and immunostimulatory cell surface adhesin-encoding EPA1 gene, respectively. Consistently, EPA1 overexpression and CgMT-C deletion elevated IL-1β (pro-inflammatory cytokine) production and diminished Cg proliferation in macrophages. Further, Cgsnf2Δ had higher Epa1 surface expression, and evoked increased IL-1β secretion, and was killed in macrophages. Akt-, p38-, NF-κB- or NLRP3 inflammasome-inhibition partially reversed increased IL-1β secretion in Cgsnf2Δ-infected macrophages. Importantly, macrophages responded to multiple Candida pathogens via NF-κB-dependent IL-1β production, underscoring NF-κB signaling's role in fungal diseases. Altogether, our findings directly link the nucleosome positioning-based chromatin remodeling to fungal immunomodulatory molecule expression.

Language models have been successfully used to model natural signals, such as images, speech, and music. A key component of these models is a high quality neural compression model that can compress high-dimensional natural signals into lower dimensional discrete tokens. To that end, we introduce a high-fidelity universal neural audio compression algorithm that achieves 90x compression of 44.1 KHz audio into tokens at just 8kbps bandwidth. We achieve this by combining advances in high-fidelity audio generation with better vector quantization techniques from the image domain, along with improved adversarial and reconstruction losses. We compress all domains (speech, environment, music, etc.) with a single universal model, making it widely applicable to generative modeling of all audio. We compare with competing audio compression algorithms, and find our method outperforms them significantly. We provide thorough ablations for every design choice, as well as open-source code and trained model weights. We hope our work can lay the foundation for the next generation of high-fidelity audio modeling.

BackgroundThe kala-azar elimination programme has resulted in a significant reduction in visceral leishmaniasis (VL) cases across the Indian Subcontinent. To detect any resurgence of transmission, a sensitive cost-effective surveillance system is required. Molecular xenomonitoring (MX), detection of pathogen DNA/RNA in vectors, provides a proxy of human infection in the lymphatic filariasis elimination programme. To determine whether MX can be used for VL surveillance in a low transmission setting, large numbers of the sand fly vector Phlebotomus argentipes are required. This study will determine the best method for capturing P. argentipes females for MX.Methodology/principal findingsThe field study was performed in two programmatic and two non-programmatic villages in Bihar, India. A total of 48 households (12/village) were recruited. Centers for Disease Control and Prevention light traps (CDC-LTs) were compared with Improved Prokopack (PKP) and mechanical vacuum aspirators (MVA) using standardised methods. Four 12x12 Latin squares, 576 collections, were attempted (12/house, 144/village,192/method). Molecular analyses of collections were conducted to confirm identification of P. argentipes and to detect human and Leishmania DNA. Operational factors, such as time burden, acceptance to householders and RNA preservation, were also considered. A total of 562 collections (97.7%) were completed with 6,809 sand flies captured. Females comprised 49.0% of captures, of which 1,934 (57.9%) were identified as P. argentipes. CDC-LTs collected 4.04 times more P. argentipes females than MVA and 3.62 times more than PKP (p<0.0001 for each). Of 21,735 mosquitoes in the same collections, no significant differences between collection methods were observed. CDC-LTs took less time to install and collect than to perform aspirations and their greater yield compensated for increased sorting time. No significant differences in Leishmania RNA detection and quantitation between methods were observed in experimentally infected sand flies maintained in conditions simulating field conditions. CDC-LTs were favoured by householders.Conclusions/significanceCDC-LTs are the most useful collection tool of those tested for MX surveillance since they collected higher numbers of P. argentipes females without compromising mosquito captures or the preservation of RNA. However, capture rates are still low.

Magnesium (Mg) shows excellent potential for orthopedic implant applications owing to its equivalent mechanical properties compared to cortical bone and its biocompatibility. However, the rapid degradation rate of magnesium and its alloys in the physiological environment results in losing their mechanical integrity before complete bone healing. In light of this, friction stir processing (FSP), a solid-state process, is used to fabricate Hopeite (Zn(PO4)2.4H2O) reinforced novel magnesium composite. As a result of the novel composite fabricated by FSP, grain refinement of the matrix phase occurs significantly. The samples were immersed in simulated body fluid (SBF) for in-vitro bioactivity and biodegradability tests. The corrosion behavior of pure Mg, FSP Mg, and FSP Mg-Hopeite composite samples was compared using electrochemical and immersion tests in SBF. It found that Mg-Hopeite composite has better corrosion resistance than FSP Mg and pure Mg. Because of grain refinement and the presence of secondary phase Hopeite in the composite, the mechanical properties and corrosion resistance improved. The bioactivity test was performed in the SBF environment, and a rapid apatite layer was formed on the surface of Mg-Hopeite composite samples during the test. Osteoblast-like MG63 cells were exposed to samples, and the MTT assay confirmed the non-toxicity of the FSP Mg-Hopeite composite. The wettability of the Mg-Hopeite composite was improved than pure Mg. The present research findings showed that the novel Mg-Hopeite composite fabricated by FSP is a promising candidate for orthopedic implant applications, unreported in the literature.

Immune evasion is critical for fungal virulence. However, how the human opportunistic pathogen Candida glabrata ( Cg ) accomplishes this is unknown. Here, using micrococcal nuclease-sequencing, RNA-sequencing, macrophage-signalling and genetic analyses, we demonstrate that chromatin reorganization in macrophage-internalized Cg , via CgSnf2 (ATPase subunit of the SWI/SNF chromatin remodelling complex), leads to upregulation and downregulation of immunosuppressive seven mannosyltransferase-cluster ( CgMT-C ) and immunostimulatory cell surface adhesin EPA1 genes, respectively. Consistently, EPA1 overexpression and CgMT-C deletion led to increased IL-1β (pro-inflammatory cytokine) production and reduced Cg proliferation in macrophages. Further, CgSNF2 deletion evoked increased IL-1β secretion, and the consequent killing of macrophage-internalized Cg , with elevated IL-1β levels being partially reversed in Akt-, p38-, NF-κB- or NLRP3 inflammasome-inhibited macrophages. Importantly, macrophages respond to multiple Candida pathogens via NF-κB-dependent IL-1β production, underscoring NF-κB signalling’s role in fungal diseases. Finally, we present the first genome-wide nucleosome map of macrophage-internalized Cg consisting of ∼12,000 dynamic and 70,000 total nucleosomes. Altogether, our findings directly link the nucleosome positioning-based chromatin remodelling to fungal immunomodulatory molecule expression, which dictates Cg fate in host immune cells.

Magnesium (Mg) shows excellent potential for orthopedic implant applications owing to its equivalent mechanical prop-erties compared to cortical bone and its biocompatibility. However, the rapid degradation rate of magnesium and its alloys in the physiological environment results in losing their mechanical integrity before complete bone healing. In light of this, friction stir processing (FSP), a solid-state process, is used to fabricate Hopeite (Zn(PO4)(2).4H(2)O) reinforced novel magnesium composite. As a result of the novel composite fabricated by FSP, grain refinement of the matrix phase occurs significantly. The samples were immersed in simulated body fluid (SBF) for in-vitro bioactivity and biodegradability tests. The corrosion behavior of pure Mg, FSP Mg, and FSP Mg-Hopeite composite samples was compared using electro-chemical and immersion tests in SBF. It found that Mg-Hopeite composite has better corrosion resistance than FSP Mg and pure Mg. Because of grain refinement and the presence of secondary phase Hopeite in the composite, the mechani-cal properties and corrosion resistance improved. The bioactivity test was performed in the SBF environment, and a rapid apatite layer was formed on the surface of Mg-Hopeite composite samples during the test. Osteoblast-like MG63 cells were exposed to samples, and the MTT assay confirmed the non-toxicity of the FSP Mg-Hopeite composite. The wettability of the Mg-Hopeite composite was improved than pure Mg. The present research findings showed that the novel Mg-Hopeite composite fabricated by FSP is a promising candidate for orthopedic implant applications, unreported in the literature.

This study investigated the sources of economic fluctuation in the Indian economy. To assess this objective, time-based and frequency-based filters were applied to extract the business cycle from the Gross Domestic Product. Further, a causal link between the business cycle and its different sources was explored using Markov's regime-switching regression. The results indicated that total factor productivity, oil supply, and monetary policy increased business cycle volatility. Furthermore, although the fiscal policy remained unaffected, trade increased economic fluctuations during the pro-market regime. The findings suggested that the views of the real business cycle and monetarist schools of thought hold true for economic fluctuation in India, as opposed to the Keynesian view.

The mechanical properties of magnesium and its alloys are remarkably close to those of natural bone, and they are biodegradable, thus they have been studied for use in consumable bio-implant applications. However, their applicability is constrained by the unpredictable corrosion rate and the moderate bioactivity of Mg-based materials in a biological environment. Considering this, a Hopeite-reinforced magnesium composite was fabricated using friction stir processing (FSP). Along with the many other advantages of FSP, it significantly reduces grain size. The impact of reduced grain size and the presence of Hopeite particles on the control of magnesium degradation was studied. After FSP, there was a significant reduction in grain size owing to severe plastic deformation and dynamic recyclization. The effect of the number of passes was studied using samples with one pass, two passes, and three passes of Mg-Hopeite composite. There was a correlation between increasing the number of FSP passes and improvements in the material's mechanical characteristics, corrosion resistance, and bioactivity behavior. The samples were immersed in simulated body fluid for in vitro bioactivity studies, where it was determined that the 3 passes FSP Mg-Hopeite composite had the greatest Ca/P ratio and hence the best bioactivity, outperforming the 2 passes composite, the 1 pass composite, and pure Mg. Apatite layer formation in FSP Mg-Hopeite composite samples has also conformed to their bioactive behavior. Electrochemical and immersion studies designed to evaluate the corrosion behavior of the material in simulated bodily fluid revealed that grain refinement and an increase in biomineralization led to an improvement in the material's corrosion resistance. Considering the findings, which indicate that 3 passes of FSP Mg-Hopeite composite possess superior corrosion resistance, and bioactive and mechanical properties in comparison to either 1 pass and 2 passes of FSP Mg-Hopeite composite or pure Mg, this material is ideally suited for use as temporary implants for biomedical applications.

Additive manufacturing (AM) is the method of fabrication of items through the sequential addition of material. It differs from subtractive methods like machining where the material is removed until a final shape is attained. AM has turned into a highly debated topic in the last decades. This is mainly due to its short manufacturing lead times, its capability of producing parts with high geometrical complexity, suitability for low-volume production, as well as customization. AM is one of the most common forms of fabrication of composites. The most common composites through AM are metal matrix, ceramic matrix, and polymer matrix. An excellent opportunity is offered by Additive Manufacturing to fabricate complex and innovative parts using composite materials. Direct and layer-by-layer processes are used in parts fabricated by AM which differs from conventional methods. Composite materials fabricated by this method provide new opportunities in design and manufacture. This chapter provides an overview of various AM processes used for fabricating composite materials. Typical multidirectional performance has been demonstrated for the fabrication of composites by AM.

We propose Wav2CLIP, a robust audio representation learning method by distilling from Contrastive Language-Image Pre-training (CLIP). We systematically evaluate Wav2CLIP on a variety of audio tasks including classification, retrieval, and generation, and show that Wav2CLIP can outperform several publicly available pre-trained audio representation algorithms. Wav2CLIP projects audio into a shared embedding space with images and text, which enables multimodal applications such as zero-shot classification, and cross-modal retrieval. Furthermore, Wav2CLIP needs just ∼10% of the data to achieve competitive performance on downstream tasks compared with fully supervised models, and is more efficient to pre-train than competing methods as it does not require learning a visual model in concert with an auditory model. Finally, we demonstrate image generation from Wav2CLIP as qualitative assessment of the shared embedding space. Our code and model weights are open sourced and made available for further applications.

Background: Heat stress in animals can occur when ambient temperatures are high and wind speeds are low, resulting in diminished growth, poor health, or even death. The purpose of this study is to evaluate the effect of Tinospora cordifolia (TC) on anti-stress and blood biochemical parameters in heat stressed (HS) Gaddi goats. Methods: Sixteen adult male Gaddi goats were split into four groups, T0H0 (no supplement no HS, control), T1H0 (diet fed with TC stem powder @ 5% of DMI with no HS), T0H1 (no supplement with HS) and T1H1 (diet fed with TC stem powder @ 5% of DMI with HS). The study was planned in a 2×2 factorial degin, with supplemetation TC stem powder (0 per cent vs. 5 per cent) and heat stress (normal vs. heat exposure) were the key effects to be studied over the 30-d trial period. Result: There was a significant interaction (supplement×stress×period) for oxidative stress indices. At the end of the experiment, the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx) and cortisol levels were higher in the T0H1 group goats than in the T1H1 group (similar to normal levels). The blood’s hematological and biochemical characteristics were equivalent among groups. However, after a 30-d trail, Hb and PCV levels in the blood were low in the stressed group. It can be culminated that, there was no significant change in hematological and blood biochemical parameters across groups, however animals’ antioxidant status were improved.

Conditional waveform synthesis models learn a distribution of audio waveforms given conditioning such as text, mel-spectrograms, or MIDI. These systems employ deep generative models that model the waveform via either sequential (autoregressive) or parallel (non-autoregressive) sampling. Generative adversarial networks (GANs) have become a common choice for non-autoregressive waveform synthesis. However, state-of-the-art GAN-based models produce artifacts when performing mel-spectrogram inversion. In this paper, we demonstrate that these artifacts correspond with an inability for the generator to learn accurate pitch and periodicity. We show that simple pitch and periodicity conditioning is insufficient for reducing this error relative to using autoregression. We discuss the inductive bias that autoregression provides for learning the relationship between instantaneous frequency and phase, and show that this inductive bias holds even when autoregressively sampling large chunks of the waveform during each forward pass. Relative to prior state-ofthe-art GAN-based models, our proposed model, Chunked Autoregressive GAN (CARGAN) reduces pitch error by 40-60%, reduces training time by 58%, maintains a fast generation speed suitable for realtime or interactive applications, and maintains or improves subjective quality.

Miniature Impact test technique was used to investigate the plausible role of hydride platelet orientation relative to the crack plane on the fracture mechanism of the Zr–2.5%Nb alloy over a wide range of temperatures. Miniature impact samples were fabricated from spools of un-hydrided (UH), circumferential hydrided (CH) and radial hydrided (RH) materials with different combinations of crack plane normal and crack growth direction. The impact tests were carried out between − 50 to 300 °C. Both microstructural (microstructure, texture) and fractographic examinations were carried out. The extent of hydride embrittlement and temperature dependence of the impact toughness were explained in terms of the hydride platelet orientation with respect to the crack plane and solubility of hydrogen in the material at different test temperatures. Variation of impact energy with test temperature for un-hydrided, circumferential and radial hydrided samples for a CA (Radial hydride lying parallel to the crack plane) and b AC (Both circumferential and radial hydride oriented normal to crack plane)

In recent years, metallic biomaterial having properties like biodegradable, biocompatible and good mechanical properties have been more desirable for orthopaedic implant applications. Magnesium based materials are extensively used in biomedical applications owing to desirable properties. Pure Magnesium can degrade too fast and lose its mechanical integrity earlier than tissue healing in a physiological environment that limits its implant application. Mg alloys and Mg matrix composites have suitable biodegradability along with excellent mechanical strength. For Mg alloys, not all alloying elements are biocompatible which limit their applications. Mg matrix composites, as biomaterials, can produce adjustable biodegradation rate and mechanical properties in physiological environments. The present review paper is focused on recent research works on Mg matrix composites for orthopaedic implant applications.

Biodegradable implant metals such as iron (Fe), magnesium (Mg), and Zinc (Zn) and its alloys have attracted extensive interest in biomedical application. Low mechanical strength of Zn, significant slower degradation of Fe, and rapid degradation Mg impede their orthopedic implant application. Further research is going on the development of biodegradable metal matrix composite owing to best suited for biomedical applications. This paper delivers a review of biodegradable metal matrix composites based on corrosion resistance, biocompatibility, and mechanical properties as favorable implant materials for orthopedic applications.

Scientists have considered the programmed segmentation of lung areas as a fundamental pre-processing undertaking when examining pulmonary radiographs (CXRs) obviously. In this paper, a method for lung field segmentation is proposed. This is performed on MATLAB image processing coding. It improves the speed of processing and detects abnormality in the lung area. The method used is Normalized Gradient Gaussian Filter and Snake segmentation is combined to improve the performance of edge detector