Changes In Cellular Components Research Articles

Unveiling the Spatiotemporal and Dose Responses within a Single Live Cancer Cell to Photoswitchable Upconversion Nanoparticle Therapeutics Using Hybrid Hyperspectral Stimulated Raman Scattering and Transient Absorption Microscopy.

Photodynamic therapy (PDT) provides an alternative approach to targeted cancer treatment, but the therapeutic mechanism of advanced nanodrugs applied to live cells and tissue is still not well understood. Herein, we employ the hybrid hyperspectral stimulated Raman scattering (SRS) and transient absorption (TA) microscopy developed for real-time in vivo visualization of the dynamic interplay between the unique photoswichable lanthanide-doped upconversion nanoparticle-conjugated rose bengal and triphenylphosphonium (LD-UCNP@CS-Rb-TPP) probe synthesized and live cancer cells. The Langmuir pharmacokinetic model associated with SRS/TA imaging is built to quantitatively track the uptakes and pharmacokinetics of LD-UCNP@CS-Rb-TPP within cancer cells. Rapid SRS/TA imaging quantifies the endocytic internalization rates of the LD-UCNP@CS-Rb-TPP probe in individual HeLa cells, and the translocation of LD-UCNP@CS-Rb-TPP from mitochondria to cell nuclei monitored during PDT can be associated with mitochondria fragmentations and the increased nuclear membrane permeability, cascading the dual organelle ablations in cancer cells. The real-time SRS spectral changes of cellular components (e.g., proteins, lipids, and DNA) observed reflect the PDT-induced oxidative damage and the dose-dependent death pattern within a single live cancer cell, thereby facilitating the real-time screening of optimal light dose and illumination duration controls in PDT. This study provides new insights into the further understanding of drug delivery and therapeutic mechanisms of photoswitchable LD-UCNP nanomedicine in live cancer cells, which are critical in the optimization of nanodrug formulations and development of precision cancer treatment in PDT.

Synovium is a sensitive tissue for mapping the negative effects of systemic iron overload in osteoarthritis: identification and validation of two potential targets

BackgroundThe prevention and treatment of osteoarthritis (OA) pose a major challenge in its research. The synovium is a critical tissue in the systematic treatment of OA. The present study aimed to investigate potential target genes and their correlation with iron overload in OA patients.MethodsThe internal datasets for analysis included the microarray datasets GSE46750, GSE55457, and GSE56409, while the external datasets for validation included GSE12021 and GSE55235. The GSE176308 dataset was used to generate single-cell RNA sequencing profiles. To investigate the expression of the target genes in synovial samples, quantitative reverse transcription-PCR, western blotting, and immunohistochemical assay were conducted. ELISA was used to detect the levels of ferritin and Fe2+ in both serum and synovium.ResultsJUN and ZFP36 were screened from the differentially expressed genes, and their mRNA were significantly reduced in the OA synovium compared to that in normal synovium. Subsequently, complex and dynamically evolving cellular components were observed in the OA synovium. The mRNA level of JUN and ZFP36 differed across various cell clusters of OA synovium and correlated with immune cell infiltration. Moreover, ferritin and Fe2+ were significantly increased in the serum and synovium of OA patients. Further, we found that JUN elevated and ZFP36 decreased at protein level.ConclusionsThe synovium is a sensitive tissue for mapping the adverse effects of systemic iron overload in OA. JUN and ZFP36 represent potential target genes for attenuating iron overload during OA treatment. Some discrepancies between the transcription and protein levels of JUN suggest that post-transcriptional modifications may be implicated. Future studies should also focus on the roles of JUN and ZFP36 in inducing changes in cellular components in the synovium during OA pathogenesis.

Dynamics of the innate immune response profile in patients with coronary heart disease at different terms after coronary artery stenting

Atherosclerosis is accompanied by damage to the vascular endothelium of arteries followed by development of inflammatory response and formation of atherosclerotic plaques. Innate immunity is an important component of this response being the earliest non-specific key mechanism. Our objective was to perform a comprehensive assessment of the cellular link of innate immunity, and to compare the results obtained at various terms after coronary stenting.The study involved 50 patients with coronary atherosclerosis (Group 1), who had clinical indications for stenting of coronary arteries, and 20 volunteers (Group 2), who have no signs of coronary artery disease. The study of immune parameters was carried out before surgery, at 4-5, 9-10 and 28-30 days after operation (during early postoperative period), as well as 6 and 12 months after stenting, i.e. over the late post-surgical period. Phenotyping of peripheral blood monocytes and lymphocytes was performed by flow cytometry using monoclonal antibodies (Beckman Coulter, USA). Intracellular content of Granzyme B was carried out with an FC500 flow laser cytofluorimeter. Metabolic activity of neutrophils was assessed by the NBT test. Alpha defensin was determined in blood plasma by ELISA technique (Hycult Biotech, USA). Statistical analysis was performed using the Statistica 12.0 program (StatSoft, USA). Statistical significance was considered significant at p ≤ 0.05.The numbers of natural killer cells and their activity, as well as those of monocytes, were increased in patients with coronary atherosclerosis. We have also shown a suppression of antigen presentation processes, an imbalance in microbicidal activity of neutrophils, with predominant secretion of antimicrobial peptides. Over the early post-surgical period, significant changes included only decreased content of intracellular Granzyme B on days 4-5, and expression of TLR4 and HLA-DR on days 4-5 and 9-10. During the late period, the patients with coronary artery disease exhibited a significant decrease in the content of some lymphocyte subsets: CD3+CD16+, CD16+Gr+ as well as amounts of monocytes: CD14+CD282+, CD14+CD284+, CD14+CD289+, along with HBT-test activity and α-defensin contents, and increased numbers of HLA-DR-expressing monocytes.There are changes in cellular component of innate immunity, indicating persistent inflammation in patients with coronary heart disease. The dynamics of revealed changes following coronary artery stenting may reflect a lability of assessed indicators mostly over the late postoperative period, thus serving a basis for predicting the outcome of coronary stenting.

Mechanosensitive channel MscL induces non-apoptotic cell death and its suppression of tumor growth by ultrasound.

Mechanosensitive channel of large conductance (MscL) is the most thoroughly studied mechanosensitive channel in prokaryotes. Owing to its small molecular weight, clear mechanical gating mechanism, and nanopore forming ability upon opening, accumulating studies are implemented in regulating cell function by activating mechanosensitive channel of large conductance in mammalian cells. This study aimed to investigate the potentials of mechanosensitive channel of large conductance as a nanomedicine and a mechano-inducer in non-small cell lung cancer (NSCLC) A549 cells from the view of molecular pathways and acoustics. The stable cytoplasmic vacuolization model about NSCLC A549 cells was established via the targeted expression of modified mechanosensitive channel of large conductance channels in different subcellular organelles. Subsequent morphological changes in cellular component and expression levels of cell death markers are analyzed by confocal imaging and western blots. The permeability of mitochondrial inner membrane (MIM) exhibited a vital role in cytoplasmic vacuolization formation. Furthermore, mechanosensitive channel of large conductance channel can be activated by low intensity focused ultrasound (LIFU) in A549 cells, and the suppression of A549 tumors in vivo was achieved by LIFU with sound pressure as low as 0.053MPa. These findings provide insights into the mechanisms underlying non-apoptotic cell death, and validate the nanochannel-based non-invasive ultrasonic strategy for cancer therapy.

Effect of acute ultraviolet radiation on Galleria mellonella health and immunity

For humans, acute and chronic overexposure to ultraviolet (UV) radiation can cause tissue damage in the form of sunburn and promote cancer(s). The immune-modulating properties of UV radiation and health-related consequences are not well known. Herein, we used the larvae of the wax moth Galleria mellonella, to determine UV-driven changes in cellular components of innate immunity. From immune cell (haemocyte) reactivity and the production of antimicrobial factors, these insects share many functional similarities with mammalian cellular innate immunity. After exposing insects to UVA or UVB for up to two hours, we monitored larval viability, susceptibility to infection, haemolymph (blood) physiology and faecal discharge. Prolonged exposure of larvae to UVB coincided with decreased survival, enhanced susceptibility to bacterial challenge, melanin synthesis in the haemolymph, compromised haemocyte functionality and changes in faecal (bacterial) content. We contend G. mellonella is a reliable in vivo model for assessing the impact of UV exposure at the whole organism and cellular levels.

Dietary Antioxidants and their Potential Role in Human Disease Management

Abstract:Reactive oxygen species (ROS) are usually destroyed by the antioxidant defence systems in our body. ROS in low concentrations may be beneficial or even indispensable for defence against microorganisms and in intracellular signaling, but its higher amounts can trigger negative chain reactions leading to various pathological conditions. A delicate balance between antioxidants and oxidants is maintained in healthy organisms and protection against the harmful effects of ROS is thus provided. The increase in ROS leading to oxidative stress plays a key role in the development of chronic diseases like cancer, diabetes, neurodegenerative, and cardiovascular diseases. Several non-enzymatic and enzymatic antioxidant activities exist as a safeguard against ROS accumulation. Imbalance between the oxidative stress and the antioxidant defence systems causes irreversible changes in cellular components and the normal cell signaling mechanisms are disrupted. The enzymatic and non-enzymatic antioxidant defences include catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GPx), vitamin A, vitamin C (ascorbic acid), vitamin E (α-tocopherol), carotenes, flavonoids, etc. Several human pathologies have been known to be treated by the use of medicinal plants owing to their antioxidant properties. Dietary antioxidants such as vitamins, minerals, carotenes, and flavonoids, with their mechanisms of antioxidant defence are discussed in the review for the effective management of various human diseases.

Single-cell RNA sequencing reveals the cellular and molecular changes that contribute to the progression of lung adenocarcinoma.

Pure ground glass nodules (GGNs) and solid nodules (SNs) represent early and relatively late stages of lung adenocarcinoma (LUAD) in radiology, respectively. The cellular and molecular characteristics of pure GGNs and SNs have not been comprehensively elucidated. Additionally, the mechanism driving the progression of lung adenocarcinoma from pure GGN to SN in radiology is also elusive. In this study, by analyzing the single-cell transcriptomic profiles of 76,762 cells from four pure GGNs, four SNs, and four normal tissues, we found that anti-tumor immunity mediated by NK and CD8+T cells gradually weakened with the progression of LUAD and humoral immunity mediated by plasma B cells was more active in SNs. Additionally, the proliferation ability of some special epithelial cell increased during the progression process from pure GGN to SN. Furthermore, stromal cells and M2 macrophages could assist the progression of LUAD. Through comprehensive analyses, we revealed dynamic changes in cellular components and intercellular interactions during the progression of LUAD. These findings could facilitate our understanding of LUAD and discovery of novel therapeutic targets.

Single-cell RNA sequencing depicts the local cell landscape in thyroid-associated ophthalmopathy

SummaryThere is a specific reactivity and characteristic remodeling of the periocular tissue in thyroid-associated ophthalmopathy (TAO). However, local cell changes responsible for these pathological processes have not been sufficiently identified. Here, single-cell RNA sequencing is performed to characterize the transcriptional changes of cellular components in the orbital connective tissue in individuals with TAO. Our study shows that lipofibroblasts with RASD1 expression are highly involved in inflammation and adipogenesis during TAO. ACKR1+ endothelial cells and adipose tissue macrophages may engage in TAO pathogenesis. We find CD8+CD57+ cytotoxic T lymphocytes with the terminal differentiation phenotype to be another source of interferon-γ, a molecule actively engaging in TAO pathogenesis. Cell-cell communication analysis reveals increased activity of CXCL8/ACKR1 and TNFSF4/TNFRSF4 interactions in TAO. This study provides a comprehensive local cell landscape of TAO and may be valuable for future therapy investigation.

Single cell Raman spectroscopy to identify different stages of proliferating human hepatocytes for cell therapy

BackgroundCell therapy provides hope for treatment of advanced liver failure. Proliferating human hepatocytes (ProliHHs) were derived from primary human hepatocytes (PHH) and as potential alternative for cell therapy in liver diseases. Due to the continuous decline of mature hepatic genes and increase of progenitor like genes during ProliHHs expanding, it is challenge to monitor the critical changes of the whole process. Raman microspectroscopy is a noninvasive, label free analytical technique with high sensitivity capacity. In this study, we evaluated the potential and feasibility to identify ProliHHs from PHH with Raman spectroscopy.MethodsRaman spectra were collected at least 600 single spectrum for PHH and ProliHHs at different stages (Passage 1 to Passage 4). Linear discriminant analysis and a two-layer machine learning model were used to analyze the Raman spectroscopy data. Significant differences in Raman bands were validated by the associated conventional kits.ResultsLinear discriminant analysis successfully classified ProliHHs at different stages and PHH. A two-layer machine learning model was established and the overall accuracy was at 84.6%. Significant differences in Raman bands have been found within different ProliHHs cell groups, especially changes at 1003 cm−1, 1206 cm−1 and 1440 cm−1. These changes were linked with reactive oxygen species, hydroxyproline and triglyceride levels in ProliHHs, and the hypothesis were consistent with the corresponding assay results.ConclusionsIn brief, Raman spectroscopy was successfully employed to identify different stages of ProliHHs during dedifferentiation process. The approach can simultaneously trace multiple changes of cellular components from somatic cells to progenitor cells.

A BRIEF DESCRIPTION OF TOMATO LEAF CURL VIRUS AND ITS MANAGEMENT

The tomato crop is affected by various biotic and abiotic factors. Among the biotic factors, Tomato Leaf Curl Virus (TLCV) is one of the most devastating viruses of tomato. The virus belongs to Begomovirus and family Gemniniviridae. The genome of virus is single-stranded-DNA. It is naturally transmitted through whitefly (Bemisia tabaci). The virus is responsible for causing severe economic losses up to 80% in tomato crop-growing regions and continuously threatening the tomato crops. The increasing severity of TLCV has compelled plant virologists to develop accurate detection and identification of the virus. TLCV is responsible for certain biochemical changes and considerably affecting the plant growth. Tomato Leaf Curl Virus is significantly influenced physiological parameters and metabolic activities of all tomato varieties. The biochemical changes of cellular components are caused due to morphological deviation of infected virus plants, and symptoms expressed by the plants can determine the magnitude of the vegetable losses. It is evident from the previous reports, that nutrients were effective in reducing TLCV from a tomato plant. Moreover, the usage of resistant varieties and controlling of whitefly population and management of TLCV by different nutrients are some of the economical and recommendable ways to manage TLCV. Among the organic management strategies, the compost is also considered best in suppressing disease severity. Moreover, epidemiology plays a vital role in the spread and progress of the disease. Keeping in view the importance of TLCV and its effects on tomato crop, the present review will highlight the etiology, physiology and management of TLCV.

Response of immature rats to a low dose of nanoparticulate silver: Alterations in behavior, cerebral vasculature-related transcriptome and permeability

The increasing production and use of silver nanoparticles (AgNPs) as antimicrobial agents in medicinal and commercial products creates a substantial risk of exposure, especially for infants and children. Our current knowledge concerning the impact of AgNPs on developing brain is insufficient. Therefore we investigated the temporal profile of transcriptional changes in cellular components of the neurovascular unit in immature rats exposed to a low dose of AgNPs. The behavior of animals under these conditions was also monitored. Significant deposition of AgNPs in brain of exposed rats was identified and found to persist over the post-exposure time. Substantial changes were noted in the transcriptional profile of tight junction proteins such as occludin and claudin-5, and pericyte-related molecules such as angiopoietin-1. Moreover, downregulation of platelet-derived growth factor (PDGFβ) and its receptor (PDGFβR) which constitute the main signaling pathway between endothelial cells and pericytes was observed. These were long-lasting effects, accompanied by overexpression of astroglial-specific GFAP mRNA and endothelial cell adhesion molecule, ICAM-1, involved in the pathomechanism of neuroinflammation. The profile of changes indicates that even low doses of AgNPs administered during the early stage of life induce dysregulation of neurovascular unit constituents which may lead to disintegration of the blood-brain barrier. This was confirmed by ultrastructural analysis that revealed enhanced permeability of cerebral microvessels resulting in perivascular edema. Changes in the behavior of exposed rats indicating pro-depressive and anti-anxiety impacts were also identified. The results show a high risk of using AgNPs in medical and consumer products dedicated for infants and children.

Cytokine storm and COVID-19: a chronicle of pro-inflammatory cytokines.

Coronavirus disease 2019 (COVID-19) has swept the world, unlike any other pandemic in the last 50 years. Our understanding of the disease has evolved rapidly since the outbreak; disease prognosis is influenced mainly by multi-organ involvement. Acute respiratory distress syndrome, heart failure, renal failure, liver damage, shock and multi-organ failure are strongly associated with morbidity and mortality. The COVID-19 disease pathology is plausibly linked to the hyperinflammatory response of the body characterized by pathological cytokine levels. The term ‘cytokine storm syndrome’ is perhaps one of the critical hallmarks of COVID-19 disease severity. In this review, we highlight prominent cytokine families and their potential role in COVID-19, the type I and II interferons, tumour necrosis factor and members of the Interleukin family. We address various changes in cellular components of the immune response corroborating with changes in cytokine levels while discussing cytokine sources and biological functions. Finally, we discuss in brief potential therapies attempting to modulate the cytokine storm.

Nanotechnology in Preclinical and Clinical Drug Development

Nanotechnology is generating a strong impact in preclinical and clinical drug development. The diversity of current nanotechnologies offers a broad platform used to enhance the performance of drug discovery screening, to develop sensitive and specific methods used to unveil the mechanisms behind the actions of drugs, to determine the function and interaction between molecules, and to study the physiological and pathological changes of cellular components. In addition, advancements in nanobiotechnology have led to the design of new nanomaterial-based drug candidates that present a novel approach to medical diagnostics and therapeutics. The biocompatible nanoarchitecture of the marketed nanocarriers used for drug delivery has increased the solubility and effectiveness of classical drugs, and has provided the technology required for the targeted delivery of encapsulated tissue-organ specific therapeutics. Because of its effect on drug development, nanotechnology serves as the foundation for many future medical endeavors. This article provides an overview of the basics of nanobiotechnology, and discusses its applications in drug discovery, design, and delivery systems.

GBIQ: a non-arbitrary, non-biased method for quantification of fluorescent images.

Non-arbitrary and non-biased quantification of fluorescent images is an essential tool for the data-centric approach to biological systems. Typical application is high-content analysis, where various phenotypic changes in cellular components and/or morphology are measured from fluorescent image data. A standard protocol to detect cellular phenotypes is cell-segmentation, in which boundaries of cellular components, such as cell nucleus and plasma membrane, are first identified to define cell segments, then acquiring various phenotypic data of each segment. To achieve reliable outcome, cell-segmentation requires manual adjustments of many parameters; this requirement could hamper automated image processing in high-throughput workflow, whose quantification must be non-arbitrary and non-biased. As a practical alternative to the segmentation-based method, we developed GBIQ (Grid Based Image Quantification), which allows comparison of cellular information without identification of single cells. GBIQ divides an image with tiles of fixed size grids and records statistics of the grids with their location coordinates, minimizing arbitrary intervenes. GBIQ requires only one parameter (size of grid) to be set; nonetheless it robustly produces results suitable for further statistical evaluation. The simplicity of GBIQ allows it to be readily implemented in an automated high-throughput image analysis workflow.

Statistical Mechanical Framework for Predicting Cellular Responses from Single-Cell Data

Developments in single-cell analysis techniques have provided detailed resolution on cellular component copy number and variation within a cell population. These data provide a probability distribution for all possible states of the cell, as determined by the measured component copy number per cell. We have developed a statistical mechanical framework that uses single-cell protein population data to model the effective interactions of the measured proteins. This method enables us to predict ensemble changes in cellular components under a variety of perturbations. This framework uses Bayesian inference to compare given models or descriptions of the system, as well as determine the uncertainty in the parameter estimations given the data. We have applied this methodology to study the impact of changes in oxygen partial pressure on the behavior of glioblastoma multiforme (GBM) cancer cells. Our framework shows how we can determine the coupling of oxygen concentration changes to the measured proteins and their underlying effective interactions.

The Protective Effect of Adiponectin and other Inflammatory Cytokines Induced by Early Obesity in Rat Heart

Objective: Obesity is linked to inflammatory processes and an imbalance in proand anti-inflammatory cytokines. A previous study revealed that early obesity causes increased cellular turnover and compensatory up-regulation of AT2 receptors in the rat heart. We surveyed the effect of early obesity on the imbalance of inflammatory cytokines according to changes in cellular components. Material and Methods: Early obesity was induced by overfeeding rat pups for 28 days. The normal litter (NL) had 10 male pups and the small litter (SL) had 3 male pups per dam. We measured body weight, serum glucose, and Masson trichrome (MT) stain. Western blotting to detect inflammatory cytokines was performed using antibodies against metalloproteinase 9 (MMP-9), tissue inhibitor of MMPs type 1 (TIMP-1) and adipokines, including adiponectin, plasminogen activator inhibitor 1 (PAI-1), and tissue necrosis factor-α (TNF-α), and monocyte chemotactic protein 1 (MCP-1) and an anti-CD68 antibody of tissue macrophages (ED-1). Results: Body weight and heart weight were significantly increased, but relative heart weight (heart weight / body weight) was significantly decreased in the SL (P < 0.001*). Both collagen and glucose in the SL were increased, but these increases were not statistically significant. Protein expression of MMP-9 (P < 0.001*), PAI-1 (P < 0.001*), and adiponectin (P = 0.009) was increased, but changes in TIMP-1, TNF-α, ED-1, and MCP-1 were not significant in the SL. (all data are 95% confidence interval) Conclusions: Early obesity induces increases in collagen and inflammatory cytokines. However, the anti-inflammatory effects of adiponectin protect the heart in early obesity and those effects of adiponectin should be investigated.

Blood-Brain Barrier Disruption Induced by Chronic Sleep Loss: Low-Grade Inflammation May Be the Link.

Sleep is a vital phenomenon related to immunomodulation at the central and peripheral level. Sleep deficient in duration and/or quality is a common problem in the modern society and is considered a risk factor to develop neurodegenerative diseases. Sleep loss in rodents induces blood-brain barrier disruption and the underlying mechanism is still unknown. Several reports indicate that sleep loss induces a systemic low-grade inflammation characterized by the release of several molecules, such as cytokines, chemokines, and acute-phase proteins; all of them may promote changes in cellular components of the blood-brain barrier, particularly on brain endothelial cells. In the present review we discuss the role of inflammatory mediators that increase during sleep loss and their association with general disturbances in peripheral endothelium and epithelium and how those inflammatory mediators may alter the blood-brain barrier. Finally, this manuscript proposes a hypothetical mechanism by which sleep loss may induce blood-brain barrier disruption, emphasizing the regulatory effect of inflammatory molecules on tight junction proteins.

Dynamic transcription profiles of "Qinguan" apple (Malus × domestica) leaves in response to Marssonina coronaria inoculation.

Marssonina apple blotch, caused by the fungus Marssonina coronaria, is one of the most destructive apple diseases in China and East Asia. A better understanding of the plant's response to fungi during pathogenesis is urgently needed to improve plant resistance and to breed resistant cultivars. To address this, the transcriptomes of “Qinguan” (a cultivar with high resistance to M. coronaria) apple leaves were sequenced at 12, 24, 48, and 72 h post-inoculation (hpi) with Marssonina coronaria. The comparative results showed that a total of 1956 genes were differentially expressed between the inoculated and control samples at the 4 time points. Gene ontology (GO) term enrichment analysis of differentially expressed genes (DEGs) revealed changes in cellular component, secondary metabolism including chalcone isomerase activity, phytoalexin biosynthetic process, anthocyanin-containing compound biosynthetic process, lignin biosynthetic process, positive regulation of flavonoid biosynthetic process; and molecular functions or biological processes related to the defense response, biotic stimulus response, wounding response and fungus response. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that DEGs were significantly enriched in flavonoid biosynthesis, vitamin B6 metabolism, phenylpropanoid biosynthesis, and the stilbenoid, diarylheptanoid and gingerol biosynthesis pathways. Furthermore, the importance of changes in cellular components and partial polyphenol compounds when encountering M. coronaria are discussed.

Application of multi-omics techniques for bioprocess design and optimization in chinese hamster ovary cells.

Significant improvements in the productivity and quality of therapeutic proteins produced in Chinese hamster ovary (CHO) cells have been reported since their establishment as host cells for biopharmaceutical production. Initial advances in the field focused on engineering strategies to manipulate genes associated with proliferation, apoptosis, and various metabolic pathways. Process engineering efforts to optimize culture media, batch-feeding strategies and culture conditions, including temperature and osmolarity, were also reported. More recently, focus has shifted toward enhancing process consistency and product quality using systems biology quality by design-based approaches during process development. Integration of different data generated using omics technologies, such as genomics, transcriptomics, proteomics and metabolomics, has facilitated a greater understanding of CHO cell biology. These techniques have enabled the provision of global information on dynamic changes in cellular components associated with different phenotypes. Using systems biology to understand these important host cells at the cellular level will undoubtedly result in further progression in the development and expression of biopharmaceutical products in CHO cells.

Infrared spectroscopy: Methods for investigating cellular components of phytopathogenic fungi response to temperature stress

Fourier-transform infrared (FTIR) spectroscopy is a sensitive method for the investigation of fungi biochemical composition changes in cells. FTIR spectroscopy can be used to study fungal cell biology by fingerprinting varieties of carbohydrates, proteins, and lipids at about 6 µm spatial resolution. FTIR can distinguish fungal changes during stressed conditions, exhibit dramatic biochemical changes without obvious morphological effects.The objective of this study was to test the hypothesis that the temperature stress, as one of the most important environmental conditions, plays a critical influence on fungal cellular components involved in pathogenic mechanisms. In the current study, we investigated the impact of temperature on the changes of cellular components of phytopathogenic fungi,Sclerotium rolfsii and Colletotrichum gloeosporioides using FTIR spectroscopy with respect to the lipid, protein and polysaccharide composition. Our results demonstrate that fungal growth of S. rolfsii and C. gloeosporioides obviously increased at temperature of32°C. The culture of S. rolfsii and C. gloeosporioides grown at 32°C shows higher content of the lipid as shown in the spectral regions of CH stretching and bending bands when compared with those of the fungi culture under temperature at 28°C, indicating that all these indicators played a role in temperature stress condition in S. rolfsii and C. gloeosporioides, probably due to enhanced activity of the fungal metabolism pathway and cell wall protection. Our findings illustrate that temperature stress caused increase biomass and the lipid composition that might be these key biochemical composition help phytopathogenic fungi S. rolfsii and C. gloeosporioides cope for survival at higher temperature. FTIR spectroscopy can be used as a new and sensitive tool to examine the biochemical changes within the fungal cell. This technique allows us to reveal structural chemical makeup and features within the different fungal growing conditions. Key words: Sclerotium rolfsii; Colletotrichum gloeosporioides, temperature response,cellular composition, FTIR spectroscopy.