Influence of Murraya koenigii extract on diabetes induced rat brain aging.

Food supplements are used to improve cognitive functions in age-related dementia. This study was designed to determine the Murraya koenigii leaves' effect on Alloxan-induced cognitive impairment in diabetic rats and the contents of oxidative stress biomarkers, catalase, reduced glutathione, and glutathione reductase in brain tissue homogenates. Wistar rats were divided into seven groups (six rats per group). Group I received saline water (1 ml, p.o.), Diabetes was induced in Groups II-VII with Alloxan (120 mg/kg/p.o). Group III was provided with Donepezil HCl (2.5 mg/kg/p.o.), Group IV, V, VI, and VII with Murraya koenigii ethanol extract (200 and 400 mg/kg/p.o.) and aqueous extract (200 and 400 mg/kg/p.o.), respectively, for 30 days. Behavior, acetylcholinesterase (AChE) activity, oxidative stress status, and histopathological features were determined in the hippocampus and cerebral cortex. Administration of Murraya koenigii ethanolic and aqueous extracts significantly (P<0.05, P<0.001) increased the number of holes crossed by rats from one chamber to another. There was an increase in the (1) latency to reach the solid platform, (2) number of squares traveled by rats on the 30th day, and (3) percentage of spontaneous alternation behavior compared to the control group. Administration for successive days markedly decreased AChE activity (P<0.05), decreased TBARS level, and increased catalase, GSH, and GR levels. Murayya koenigii could be a promising food supplement for people with dementia. However, more research into sub-chronic toxicity and pharmacokinetic and pharmacodynamics interactions is essential.

Characterizing the Macrophage Population in Patients With Idiopathic Subglottic Stenosis.

OBJECTIVES: Idiopathic subglottic stenosis (iSGS) is characterized by progressive fibrosis and subglottic luminal narrowing. Currently, immune characterization has focused on T-cells; however, macrophages remain largely unexplored. The goals of this study are to characterize the transcriptome of iSGS macrophages and the fibrogenic nature of identifed biomarkers. STUDY DESIGN: Bioinformatics and in vitro. METHODS: Human tracheal biopsies from iSGS scar (n = 4), and matched non-scar (n = 4) regions were analyzed using single-cell RNA-seq (scRNA-seq). Immunofluorescence (IF) was performed on rapidly processed autopsies (RPA) and iSGS tracheal resections (n = 4) to co-localize S100A8/9 and CD11b. Collagen gene/protein expression was assessed in iSGS fibroblasts (n = 4) treated with protein S100A8/9 (1000 ng/ml). Macrophages were subclustered to identify distinct subpopulations. RESULTS: scRNA-seq analysis revealed S100A8/S100A9 (fold change (FC) = 4.1/1.88, p < 0.001) as top differentially expressed genes in iSGS macrophages. IF exhibited increased CD11b+/S100A8/9+ cells in tracheal samples of iSGS versus RPA (26.75% ± 7.08 vs. 0.594% ± 0.974, n = 4, p = 0.029). iSGS fibroblasts treated with S100A8/9 demonstrated increased gene expression of COL1A1 (FC = 2.30 ± 0.45, p = 0.03, n = 4) and COL3A1 (FC = 2.44 ± 0.40, p = 0.03, n = 4). COL1A1 protein assays revealed an increase in the experimental group, albeit not significant, (p = 0.12, n = 4). Finally, macrophage sub clustering revealed one subpopulation as a predominant source of S100A8/S100A9 expression (FC = 7.94/5.47, p < 0.001). CONCLUSIONS: S100A8/9 is a key biomarker in iSGS macrophages. Although S100A8/9 demonstrates profibrotic nature in vitro, the role of S100A8/9+ macrophages in vivo warrants further investigation. LEVEL OF EVIDENCE: NA Laryngoscope, 133:2308-2316, 2023.

RBM46 is essential for gametogenesis and functions in post-transcriptional roles affecting meiotic cohesin subunits

RBM46 is a germ cell-specific RNA-binding protein required for gametogenesis, but the targets and molecular functions of RBM46 remain unknown. Here, we demonstrate that RBM46 binds at specific motifs in the 3'UTRs of mRNAs encoding multiple meiotic cohesin subunits and show that RBM46 is required for normal synaptonemal complex formation during meiosis initiation. Using a recently reported, high-resolution technique known as LACE-seq and working with low-input cells, we profiled the targets of RBM46 at single-nucleotide resolution in leptotene and zygotene stage gametes. We found that RBM46 preferentially binds target mRNAs containing GCCUAU/GUUCGA motifs in their 3'UTRs regions. In Rbm46 knockout mice, the RBM46-target cohesin subunits displayed unaltered mRNA levels but had reduced translation, resulting in the failed assembly of axial elements, synapsis disruption, and meiotic arrest. Our study thus provides mechanistic insights into the molecular functions of RBM46 in gametogenesis and illustrates the power of LACE-seq for investigations of RNA-binding protein functions when working with low-abundance input materials.

Artificial intelligence, BI-RADS evaluation and morphometry: A novel combination to diagnose breast cancer using ultrasonography, results from multi-center cohorts.

PURPOSE: To develop and validate a machine learning (ML) model for the classification of breast lesions on ultrasound images. METHOD: In the present study, three separate data cohorts containing 1288 breast lesions from three countries (Malaysia, Iran, and Turkey) were utilized for MLmodel development and external validation. The model was trained on ultrasound images of 725 breast lesions, and validation was done separately on the remaining data. An expert radiologist and a radiology resident classified the lesions based on the BI-RADS lexicon. Thirteen morphometric features were selected from a contour of the lesion and underwent a three-step feature selection process. Five features were chosen to be fed into the model separately and combined with the imaging signs mentioned in the BI-RADS reference guide. A support vector classifier was trained and optimized. RESULTS: The diagnostic profile of the model with various input data was compared to the expert radiologist and radiology resident. The agreement of each approach with histopathologic specimens was also determined. Based on BI-RADS and morphometric features, the model achieved an area under the receiver operating characteristic (ROC) curve (AUC) of 0.885, which is higher than the expert radiologist and radiology resident performances with AUC of 0.814 and 0.632, respectively in all cohorts. DeLong's test also showed that the AUC of the ML protocol was significantly different from that of the expert radiologist (ΔAUCs = 0.071, 95%CI: (0.056, 0.086), P = 0.005). CONCLUSIONS: These results support the possible role of morphometric features in enhancing the already well-excepted classification schemes.

An integration study of microalgae bioactive retention: From microalgae biomass to microalgae bioactives nanoparticle.

Microalgae metabolites include biologically active compounds with therapeutic effects such as anticancer, anti-inflammatory and immunomodulation effects. One of the most recent focuses is on utilizing microalgae lipid-based biologically active compounds in food applications. However, most microalgae biological active compounds in their natural forms have common drawbacks like low solubility, low physicochemical stability and strong susceptibility to degradation, which significantly limits their application in foods, therefore, it is important to find solutions to retain their functional properties. In the present work, a comprehensive review on multi-product biorefinery was carried out from upstream processing stage to downstream processing stage, and identify critical processes and factors that impact bioactive material acquisition and retention. Furthermore, since nanoencapsulation technology emerges as an effective solution for microalgae nutraceutical product's retention, this work also focus on the nanoparticle perspective and comprehensively reviews the current nanoencapsulation solutions of the microalgae bioactive extract products. The aim is to depict advances in the formulations of microalage bioactive nanoparticles and provide a critical analysis of the reported nanoparticle formation. Overall, through the investigation of microalgae from biomass to bioactive nanoparticles, we aim to facilitate microalgae nutraceuticals incorporation as high value-added ingredients in more functional food that can improve human health.

Cystic adventitial disease of the popliteal vein and artery in siblings.

Cystic adventitial disease (CAD) is an uncommon condition in which mucoid cysts form within the adventitial layer of the arterial or venous wall. We have presented two cases in which two first-degree relatives (brother and sister) had acquired CAD ∼6 years apart, one involving the popliteal artery and the other involving the popliteal vein. We have reported a rare case of a possible familial association of CAD. We have discussed the etiology, diagnostic criteria, and imaging modalities between arterial and venous CAD to aid in the management and selection of optimal treatment strategies.

The Evaluation of Drug Delivery Nanocarrier Development and Pharmacological Briefing for Metabolic-Associated Fatty Liver Disease (MAFLD): An Update.

Current research indicates that the next silent epidemic will be linked to chronic liver diseases, specifically non-alcoholic fatty liver disease (NAFLD), which was renamed as metabolic-associated fatty liver disease (MAFLD) in 2020. Globally, MAFLD mortality is on the rise. The etiology of MAFLD is multifactorial and still incompletely understood, but includes the accumulation of intrahepatic lipids, alterations in energy metabolism, insulin resistance, and inflammatory processes. The available MAFLD treatment, therefore, relies on improving the patient's lifestyle and multidisciplinary pharmacotherapeutic options, whereas the option of surgery is useless without managing the comorbidities of the MAFLD. Nanotechnology is an emerging approach addressing MAFLD, where nanoformulations are suggested to improve the safety and physicochemical properties of conventional drugs/herbal medicines, physical, chemical, and physiological stability, and liver-targeting properties. A wide variety of liver nanosystems were constructed and delivered to the liver, only those that addressed the MAFLD were discussed in this review in terms of the nanocarrier classes, particle size, shape, zeta potential and offered dissolution rate(s), the suitable preparation method(s), excipients (with synergistic effects), and the suitable drug/compound for loading. The advantages and challenges of each nanocarrier and the focus on potential promising perspectives in the production of MAFLD nanomedicine were also highlighted.

Human Intestinal Organoids Recapitulate Enteric Infections of Enterovirus and Coronavirus.

Enteroviruses, such as EV-A71 and CVA16, mainly infect the human gastrointestinal tract. Human coronaviruses, including SARS-CoV and SARS-CoV-2, have been variably associated with gastrointestinal symptoms. We aimed to optimize the human intestinal organoids and hypothesize that these optimized intestinal organoids can recapitulate enteric infections of enterovirus and coronavirus. We demonstrate that the optimized human intestinal organoids enable better simulation of the native human intestinal epithelium, and that they are significantly more susceptible to EV-A71 than CVA16. Higher replication of EV-A71 than CVA16 in the intestinal organoids triggers a more vigorous cellular response. However, SARS-CoV and SARS-CoV-2 exhibit distinct dynamics of virus-host interaction; more robust propagation of SARS-CoV triggers minimal cellular response, whereas, SARS-CoV-2 exhibits lower replication capacity but elicits a moderate cellular response. Taken together, the disparate profile of the virus-host interaction of enteroviruses and coronaviruses in human intestinal organoids may unravel the cellular basis of the distinct pathogenicity of these viral pathogens.

SARS-CoV-2 Induces a More Robust Innate Immune Response and Replicates Less Efficiently Than SARS-CoV in the Human Intestines: An Ex Vivo Study With Implications on Pathogenesis of COVID-19.

BACKGROUND AND AIMS: Besides prominent respiratory involvement, gastrointestinal manifestations are commonly reported in Coronavirus Disease 2019 (COVID-19) patients. We compared infection of ex vivo human intestinal tissues by SARS-CoV-2 and SARS-CoV with respect to their replication kinetics and immune activation profile. METHODS: Human intestinal tissues were obtained from patients while undergoing surgical operations at Queen Mary Hospital, Hong Kong. Upon surgical removal, the tissues were immediately processed and infected with SARS-CoV-2 or SARS-CoV. Replication kinetics were determined with immunohistochemistry, qRT-PCR, and plaque assays. Immune activation in the infected intestinal tissues was assessed by detecting the gene expression of interferons and representative pro-inflammatory cytokines and chemokines. RESULTS: SARS-CoV-2 could infect and productively replicate in the ex vivo human intestinal tissues with release of infectious virus particles, but not in ex vivo human liver and kidney tissues. Importantly, SARS-CoV-2 replicated less efficiently than SARS-CoV, induced less cytopathology in the human intestinal epithelium, and induced a more robust innate immune response including the activation of both type I and type III interferons, than SARS-CoV in human intestinal tissues. CONCLUSION: Using the ex vivo human intestinal tissues as a physiologically relevant model, our data indicated that SARS-CoV-2 could productively replicate in the human gut and suggested that the gastrointestinal tract might serve as an alternative route of virus dissemination. SARS-CoV-2 replicated less efficiently and induced less cytopathology than SARS-CoV in keeping with the clinical observations reported for COVID-19 and SARS, which might be the result of a more robust immune activation by SARS-CoV-2 than SARS-CoV in the human intestine.

Biocontrol capability of bacteriophages against soft rot disease caused by Dickeya chrysanthemi

Aims@#This study aims to evaluate the effectiveness of bacteriophages isolated from Klang and Penang, Malaysia against Dickeya chrysanthemi that causes soft rot disease.@*Methodology and results@#Basic characterization such as dextrose test, citrate test, lactose fermentation test and ornithine test were carried out on D. chrysanthemi. Activity of bacteriophages against D. chrysanthemi was evaluated using spot test. Double agar overlay assay was performed to purify and enumerate the quantify of bacteriophages.Bacteriophages were also checked for its effectiveness in controlling soft rot on post-harvested vegetables: potato (Solanum tuberosum), cucumber (Cucumis sativus) and apple (Malus domestica). Results showed that D. chrysanthemiable to utilize citrate and dextrose as the source of energy, which indicated that D. chrysanthemi inclined to choose fruits and vegetables containing citrate and dextrose as the target of attack. Clear zone observed on the bacterial lawn (spot test) indicated the ability of the bacteriophages to infect and lyse D. chrysanthemi. All the bacteriophages studied herein reached the highest concentration on day 3 and were monovalent.@*Conclusion, significance and impact of study@#All the isolated bacteriophages were able to restrain the spreading of soft rot caused by D. chrysanthemi either work alone or as cocktail. This study provides information for the formulation development of bacteriophage against soft rot disease cause by D. chrysanthemi. Furthermore, this study reveals the potential of locally isolated bacteriophages against the D. chrysanthemi and paving the application of phage treatment on agriculture products that are not limited to potatoes, cucumber and apple.

Liquid and Solid Self-Emulsifying Drug Delivery Systems (SEDDs) as Carriers for the Oral Delivery of Azithromycin: Optimization, In Vitro Characterization and Stability Assessment.

Azithromycin (AZM) is a macrolide antibiotic used for the treatment of various bacterial infections. The drug is known to have low oral bioavailability (37%) which may be attributed to its relatively high molecular weight, low solubility, dissolution rate, and incomplete intestinal absorption. To overcome these drawbacks, liquid (L) and solid (S) self-emulsifying drug delivery systems (SEDDs) of AZM were developed and optimized. Eight different pseudo-ternary diagrams were constructed based on the drug solubility and the emulsification studies in various SEDDs excipients at different surfactant to co-surfactant (Smix) ratios. Droplet size (DS) < 150 nm, dispersity (D) ≤ 0.7, and transmittance (T)% > 85 in three diluents of distilled water (DW), 0.1 mM HCl, and simulated intestinal fluids (SIF) were considered as the selection criteria. The final formulations of L-SEDDs (L-F1(H)), and S-SEDDs (S-F1(H)) were able to meet the selection requirements. Both formulations were proven to be cytocompatible and able to open up the cellular epithelial tight junctions (TJ). The drug dissolution studies showed that after 5 min > 90% and 52.22% of the AZM was released from liquid and solid SEDDs formulations in DW, respectively, compared to 11.27% of the pure AZM, suggesting the developed SEDDs may enhance the oral delivery of the drug. The formulations were stable at refrigerator storage conditions.

SARS-CoV-2 detection by nasal strips: a superior tool for surveillance of pediatric populations.

BackgroundDeep throat saliva (DTS) and pooled nasopharyngeal swab and throat swab (NPSTS) are utilized for viral detection. DTS is challenging for children. Swabbing the respiratory mucosa requires trained personnel and may trigger sneezing and coughing, which generate droplets. A reliable, simple and safe sampling method applicable to a wide age range is required for community-based surveillance. MethodsWe introduced nasal strip as an easy and low-risk collection method. Asymptomatic and symptomatic SARS-CoV-2 infected patients (n = 38) were recruited. Nasal epithelial lining fluid (NELF) (n = 43) strip paired with nasal swab (n = 13) were collected by a healthcare worker to compare with NPSTS (n = 21) or DTS (n =22) collected within 24 hours as reference. All samples were subjected to viral RNA quantitation by real-time PCR targeting the nucleoprotein gene. ResultsComparable Ct values were observed between paired nasal strip and nasal swab samples. The agreement between nasal strip samples and NPSTS was 94.44% and 100% for NPSTS positive and negative samples. Higher viral RNA concentration was detected in nasal strips than DTS samples. False-negative results were recorded in six DTS specimens, of which four were from children. Storage at room temperature up to 72 (n = 3) hours did not affect diagnostic yield of nasal strips. ConclusionsNasal strip is a reliable and non-invasive sampling method for SARS-CoV-2 detection, and viral detection remains stable for at least 72 hours. It can be used as an alternative tool for community-based surveillance.

Venous congestion from brachiocephalic vein stenosis mimicking sclerotic vertebral lesions

@#In central venous obstruction, vertebral marrow enhancement (VME) may be seen secondary to collateral venous flow via the vertebral venous plexus. 1 There are only sporadic case reports on pseudolesions due to collateral enhancement mimicking sclerotic osseous metastasis. This abnormal vertebral enhancement may lead to erroneous diagnosis of sclerotic metastases or suspicious bone lesion which affect the management and prognosis. We describe a case of brachiocephalic vein obstruction-related vertebral body pseudolesions as identified in contrast-enhanced computed tomography (CECT) scan

The use of tissue fiducial markers in improving the accuracy of post-prostatectomy radiotherapy

PURPOSE: The aim of this retrospective study was to investigate the use of a radiopaque tissue fiducial marker (TFM) in the treatment of prostate cancer patients who undergo post-prostatectomy radiotherapy (PPRT). TFM safety, its role and benefit in quantifying the set-up uncertainties in patients undergoing PPRT image-guided radiotherapy were assessed. MATERIALS AND METHODS: A total of 45 consecutive PPRT patients underwent transperineal implantation of TFM at the level of vesicourethral anastomosis in the retrovesical tissue prior to intensity-modulated radiotherapy. Prostate bed motion was calculated by measuring the position of the TFM relative to the pelvic bony anatomy on daily cone-beam computed tomography. The stability and visibility of the TFM were assessed in the initial 10 patients. RESULTS: No postoperative complications were recorded. A total of 3,500 images were analysed. The calculated prostate bed motion for bony landmark matching relative to TFM were 2.25 mm in the left-right, 5.89 mm in the superior-inferior, and 6.59 mm in the anterior-posterior directions. A significant 36% reduction in the mean volume of rectum receiving 70 Gy (rV₇₀) was achieved for a uniform planning target volume (PTV) margin of 7 mm compared with the Australian and New Zealand Faculty of Radiation Oncology Genito-Urinary Group recommended PTV margin of 10 mm. CONCLUSION: The use of TFM was safe and can potentially eliminate set-up errors associated with bony landmark matching, thereby allowing for tighter PTV margins and a consequent favourable reduction in dose delivered to the bladder and rectum, with potential improvements in toxicities.

Towards precision medicine: from quantitative imaging to radiomics / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Radiology (imaging) and imaging-guided interventions, which provide multi-parametric morphologic and functional information, are playing an increasingly significant role in precision medicine. Radiologists are trained to understand the imaging phenotypes, transcribe those observations (phenotypes) to correlate with underlying diseases and to characterize the images. However, in order to understand and characterize the molecular phenotype (to obtain genomic information) of solid heterogeneous tumours, the advanced sequencing of those tissues using biopsy is required. Thus, radiologists image the tissues from various views and angles in order to have the complete image phenotypes, thereby acquiring a huge amount of data. Deriving meaningful details from all these radiological data becomes challenging and raises the big data issues. Therefore, interest in the application of radiomics has been growing in recent years as it has the potential to provide significant interpretive and predictive information for decision support. Radiomics is a combination of conventional computer-aided diagnosis, deep learning methods, and human skills, and thus can be used for quantitative characterization of tumour phenotypes. This paper discusses the overview of radiomics workflow, the results of various radiomics-based studies conducted using various radiological images such as computed tomography (CT), magnetic resonance imaging (MRI), and positron-emission tomography (PET), the challenges we are facing, and the potential contribution of radiomics towards precision medicine.

Complications and Pain in Obstructive Sleep Apnoea - Comparing Single and Multilevel Surgery

<p><b>INTRODUCTION</b>This study aimed to investigate differences in the complication rate and postoperative pain score between single and multilevel surgery performed on patients with obstructive sleep apnoea.</p><p><b>MATERIALS AND METHODS</b>A retrospective analysis was performed on patients with obstructive sleep apnoea who underwent surgery in a tertiary referral centre over 3 years. Patients who underwent single-level nasal, palatal or tongue surgery were compared with patients who underwent concurrent multilevel surgery of 2 or 3 levels. Complications and the postoperative Visual Analogue Scale pain score were recorded and the outcomes between single and multilevel groups were compared.</p><p><b>RESULTS</b>The overall complication rate for patients was 12.6%, 6.7% if only patients requiring intervention were considered. The adjusted odds ratio (OR) for complication rate for patients undergoing multilevel surgery and single-level surgery was 2.76. It was statistically significant (=0.053) after adjusting for confounders. There was more pain in patients who underwent multilevel surgery than in the single-level surgery group.</p><p><b>CONCLUSION</b>Concurrent multilevel surgery is a feasible option in patients with multilevel obstruction, especially if they are undergoing palate and tongue surgery, nose and palate surgery or nose and tongue surgery. There may be more complications, though it is not statistically significant. Further studies are required to investigate the differences between single-level nasal surgery and 3-level multilevel surgery. More patients undergoing multilevel surgery than single-level surgery experienced pain. Multilevel surgery patients should have their analgesia reviewed regularly and titrated accordingly.</p>

Conserved miR-10 family represses proliferation and induces apoptosis in ovarian granulosa cells.

Granulosa cells (GCs) are essential somatic cells in the ovary and play an important role in folliculogenesis. Brain-derived neurotropic factor (BDNF) and the TGF-ß pathway have been identified as a critical hormone and signalling pathway, respectively, in GCs. In this study, we found that a conserved microRNA family that includes miR-10a and miR-10b repressed proliferation and induced apoptosis in human, mouse, and rat GCs (hGCs, mGCs and rGCs, respectively). Moreover, essential hormones and growth factors in the follicle, such as FSH, FGF9 and some ligands in the TGF-ß pathway (TGFß1, Activin A, BMP4 and BMP15), inhibited miR-10a and miR-10b expression in GCs. In contrast, the miR-10 family suppressed many key genes in the TGF-ß pathway, suggesting a negative feedback loop between the miR-10 family and the TGF-ß pathway in GCs. By using bioinformatics approaches, RNA-seq, qPCR, FISH, immunofluorescence, Western blot and luciferase reporter assays, BDNF was identified as a direct target of the miR-10 family in GCs. Additionally, reintroduction of BDNF rescued the effects of miR-10a and miR-10b in GCs. Collectively, miR-10a and miR-10b repressed GC development during folliculogenesis by repressing BDNF and the TGF-ß pathway. These effects by the miR-10 family on GCs are conserved among different species.

Screening and identification of Trametes lactinea and Pycnoporouscoccineus isolated from Kampar for laccase production

Aims: Laccase is a blue copper oxidase that catalyses four electron reduction of molecular oxygen to water. It is able to oxidise aromatic compounds with molecular oxygen as the terminal electron acceptor. The aim of this study was to screen for laccase producing basidiomycetes isolated from decaying woods and tree trunks around Kampar, Perak. Methodology and results: The isolated basidiomycetes were screened for their laccase activity on different agar plates supplemented with 2, 2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), guaiacol and Remazol Brillant Blue R (RBBR), respectively. In the presence of laccase, the colourless ABTS and guaiacol were oxidised to form blue-green and reddish-brown coloured zone around the fungal colony, respectively; whereas the blue RBBR was decolourised by the enzyme. Colour or colourless halo zones that are formed on the agar plates indicate the presence of ligninolytic enzyme activities. Isolates KA1 and TR9 indicated the highest enzymatic hydrolysis on ABTS plates with the halo zone ratio of 1.43  0.04 and 0.98  0.01, respectively. Based on the BLAST results from the amplicon of ITS1 and ITS4 primers, Isolates KA1 and TR9 were identified as Trametes lactinea and Pycnoporous coccineus, respectively. Under submerged fermentation, P. coccineus has higher laccase production (0.72 U/mL) compared with T. lactinea (0.16 U/mL). Conclusion, significance and impact of study: Both T. lactinea and P. coccineus are potential strains for laccase production which can be used for dye decolourisation and degradation. Future studies will focus on the application of the laccase in textile dye degradation.

Novel egg white-based 3-D cell culture system.

Although three dimensional (3-D) cell culture systems have numerous advantages over traditional monolayer culture, the currently available 3-D cell culture media are cost-prohibitive for regular use by the majority of research laboratories. Here we show a simple system based on avian egg white that supports growth of cells in 3-D, at a significantly decreased cost. Specifically, we show that growth of immortalized human breast epithelial cells (MCF10A) in egg white-based medium results in formation of acini with hollow lumens, apoptotic clearance of the cells in the lumen, and apicobasal polarization comparable to what has been described using established 3-D culture media such as reconstituted basement membrane preparations (BM). There was no significant difference in MCF10A proliferation and acinar size between egg white and BM. We also cultured different established cell lines, oncogene-transformed MCF10A, and mouse mammary epithelial cells in egg white and BM, and observed similar morphology. In summary, our data convincingly argue that egg white can be used as a suitable alternative model for 3-D cell culture studies. We strongly believe that this simple and inexpensive method should allow researchers to perform 3-D cell culture experiments on a regular basis, and result in a dramatic increase of use of the 3-D cell culture in research. Thus, this finding lays the foundation for significantly increased, cost-effective use of 3-D cultures in cell biology.