Functionalized siRNA-chitosan nanoformulations promote triple-negative breast cancer cell death via blocking the miRNA-21/AKT/ERK signaling axis: in-silico and in vitro studies.

Oncogenic microRNA (miRNA), especially miRNA-21 upregulation in triple-negative breast cancer (TNBC), suggests a new class of therapeutic targets. In this study, we aimed to create GE11 peptide-conjugated small interfering RNA-loaded chitosan nanoparticles (GE11-siRNA-CSNPs) for the targeting of EGFR overexpressed TNBC and selectively inhibit miRNA-21 expression. A variety of in-silico and in vitro cellular and molecular studies were conducted to investigate the binding affinities of specific targets used as well as the anticancer efficacies and mechanisms of GE11-siRNA-CSNPs in TNBC cells. An in-silico assessment reveals a distinct binding affinity of miRNA-21 with siRNA as well as between the extracellular domain of EGFR and synthesized peptides. Notably, the in vitro results showed that GE11-siRNA-CSNPs were revealed to have better cytotoxicity against TNBC cells. It significantly inhibits miRNA-21 expression, cell migration, and colony formation. The results also indicated that GE11-siRNA-CSNPs impeded cell cycle progression. It induces cell death by reducing the expression of the antiapoptotic gene Bcl-2 and increasing the expression of the proapoptotic genes Bax, Caspase 3, and Caspase 9. Additionally, the docking analysis and immunoblot investigations verified that GE1-siRNA-CSNPs, which specifically target TNBC cells and suppress miRNA-21, can prevent the effects of miRNA-21 on the proliferation of TNBC cells via controlling EGFR and subsequently inhibiting the PI3K/AKT and ERK1/2 signaling axis. The GE11-siRNA-CSNPs design, which specifically targets TNBC cells, offers a novel approach for the treatment of breast cancer with improved effectiveness. This study suggests that GE11-siRNA-CSNPs could be a promising candidate for further assessment as an additional strategy in the treatment of TNBC.

Analysis of Socio Demographic Risk Factors of Cervical Intraepithelial Neoplasia at Colposcopic Clinic in Mymensingh Medical College Hospital.

Cervical intraepithelial neoplasia (CIN) is a pre-malignant lesion of the cervix of uterus. Several risk factors increased the risk of developing CIN. Purpose of this study was to evaluate the socio-demographic risk factors related to CIN at our setting. This Cross sectional observational study was performed at Colposcopic clinic of Mymensingh Medical College Hospital (MMCH), Mymensingh, Bangladesh from 9th November 2017 to 8th May 2018. Overall demographic features of 50 patients of precancerous cervical lesion show that, most of the patients belonged to the age group 30-39 years (46.0%), mean age was 32.7±10.3 years. Maximum numbers of respondents came from rural area (58.0%), followed by urban area (42.0%). Among them house wife- 46.0%, daily worker- 30.0% and illiterate 36.0%, primary level of education 32.6%. Among the patients the poor class 46.0% and 58.0% of the respondents were married at age ≤19 year. Among the respondents, (26.0%) were conceived their first child 1 month after their marriage and 54.0% of the women within 12 months of marriage. In this study multipara were (62.0%). Oral contraceptive pill was taken by (42.0%) of patients. The association of risk factors revealed that betel or tobacco chewing present in 28.0% cases; history of menstrual regulation (MR), dilatation, evacuation and curettage (DE&C), miscarriage were in 26.0% cases, family history of cancer were in 16.0% cases and multiple sexual exposure was in 10.0% cases. Women develop pre-malignant cervical lesions require early treatment. It is recommended that provision of proper health care support, early detection of CIN and proper management, can reduce the fatal outcome of the disease.

Comparison of Decaying Antibody Level After SARS-CoV-2 Infection in Case of Home Treated and Hospital Treated Patients at Different Interval.

The aim of the study was to evaluate the quantities difference and persistence of antibody in home and hospital treated patients at different interval. A cross-sectional study was conducted in the Department of Biochemistry and Molecular Biology, BIRDEM General Hospital, from July, 2020 to June, 2021. This study protocol was approved by Institutional Review Board (IRB), BIRDEM. SARS-CoV-2 caused death of huge number of people. The disease most commonly represents with fever, cough, loss of smell sensation, fatigue etc. Who had mild symptoms did not admit at hospital. But when the disease progress to respiratory distress, involvement of lung field more than 50.0% and other complications then the patients need hospital admission. Hospital treated patients showed higher antibody status in comparison to home treated patients. Moreover it was reported that increased amount of antibody developed after 3rd and 4th months of SARS-CoV-2 infection. In this study, statistically significant difference of decaying antibody level between home and hospital treated patients was observed.

Gingerols and shogaols: A multi-faceted review of their extraction, formulation, and analysis in drugs and biofluids to maximize their nutraceutical and pharmaceutical applications.

Gingerols represent the main bioactive compounds in ginger drugs mostly Zinigiber officinale (F. Zingebraceae) and account for the biological activities and the strong/pungent flavor in ginger. Ginger (Z. officinale) rhizome is one of the most valued herbal drugs for ailments' treatment in many ayurvedic medicine asides from its culinary applications as a spice. Gingerols and their dehydrated products shogaols are phenolic phytochemicals found in members of the Zingiberaceae family and account for most of their effects including anti-inflammatory and anticancer activities. This review entails most of the novel trends related to the extraction, optimization, and formulations of gingerols and shogaols to insure best recoveries and efficacies from their natural resources. Further, it presents a comprehensive overview of the different analytical approaches for the determination of gingerols/shogaols' levels in nutraceuticals to ensure highest quality and for their detection in body fluids for proof of efficacy.

Enzyme Immobilization Technologies and Industrial Applications.

Enzymes play vital roles in diverse industrial sectors and are essential components of many industrial products. Immobilized enzymes possess higher resistance to environmental changes and can be recovered/recycled easily when compared to the free forms. The primary benefit of immobilization is protecting the enzymes from the harsh environmental conditions (e.g., elevated temperatures, extreme pH values, etc.). The immobilized enzymes can be utilized in various large-scale industries, e.g., medical, food, detergent, textile, and pharmaceutical industries, besides being used in water treatment plants. According to the required application, a suitable enzyme immobilization technique and suitable carrier materials are chosen. Enzyme immobilization techniques involve covalent binding, encapsulation, entrapment, adsorption, etc. This review mainly covers enzyme immobilization by various techniques and their usage in different industrial applications starting from 1992 until 2022. It also focuses on the multiscale operation of immobilized enzymes to maximize yields of certain products. Lastly, the severe consequence of the COVID-19 pandemic on global enzyme production is briefly discussed.

Thinking about drinking: Acculturation and alcohol-related cognitions among college-bound Latinas.

Objective: This study examined acculturation with positive alcohol expectancies (PAE) and alcohol use intentions among college-bound Latinas using a bidimensional (ie U.S. acculturation/enculturation) and bidomain (ie behaviors/values) acculturation framework. Participants: A total of 298 Latina young adults between 18 and 20 years old were included in this analysis. Methods: Data were collected the summer before participants began college for the first time. We used an online survey to assess acculturation, PAE, and alcohol use expectancies. Results: Path analyses showed that U.S. acculturation values were related to more PAE and alcohol use intentions. U.S. acculturation behaviors were related to more alcohol use intentions, and the pathway was moderated by PAE. There was also an interaction between U.S. acculturation and enculturation behaviors predicting alcohol use intentions. Conclusion: This study sheds light on how acculturating Latina young women think about alcohol use prior to beginning college, which is an opportune window for targeted prevention programs.

Nanoencapsulated Extract of a Red Seaweed (Rhodophyta) Species as a Promising Source of Natural Antioxidants.

Marine seaweed species represent an important source of bioactive compounds possessing antioxidant activity. This study aimed at evaluating the antioxidant capacity of the Jania rubens algal extract by means of two antioxidant assays, i.e., 2,2-diphenyl-1-picrylhydrazyl and ferric-reducing antioxidant power. The seaweeds' total phenolic and flavonoid contents were also assayed as markers of antioxidant activity. To identify active agents responsible for the antioxidant activity, gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry were used for comprehensive metabolites characterization. To enhance the Jania rubens efficacy, the extract was nanoencapsulated using an ionic gelation method by means of high-pressure homogenization. The optimum nanoformulation had a particle size of 161 nm, a ζ potential of 31.2 mV, a polydispersity index of 0.211, and entrapment efficiency of 99.7%. The in vitro phytochemicals' release profiles of Jania rubens chitosan nanoparticles in comparison to the concentration of the raw algal extract were studied by the dialysis bag diffusion method revealing that the extract was released in a controlled pattern. The results indicated the potential advantages of the encapsulated Jania rubens extract, with its potent antioxidant activity, for use in different applications where sustained release is useful.

Chimeric antigen receptor-modified human regulatory T cells that constitutively express IL-10 maintain their phenotype and are potently suppressive.

Clinical trials of Treg therapy in transplantation are currently entering phases IIa and IIb, with the majority of these employing polyclonal Treg populations that harbor a broad specificity. Enhancing Treg specificity is possible with the use of chimeric antigen receptors (CARs), which can be customized to respond to a specific human leukocyte antigen (HLA). In this study, we build on our previous work in the development of HLA-A2 CAR-Tregs by further equipping cells with the constitutive expression of interleukin 10 (IL-10) and an imaging reporter as additional payloads. Cells were engineered to express combinations of these domains and assessed for phenotype and function. Cells expressing the full construct maintained a stable phenotype after transduction, were specifically activated by HLA-A2, and suppressed alloresponses potently. The addition of IL-10 provided an additional advantage to suppressive capacity. This study therefore provides an important proof-of-principle for this cell engineering approach for next-generation Treg therapy in transplantation.

Collagen I Fibrous Substrates Modulate the Proliferation and Secretome of Estrogen Receptor-Positive Breast Tumor Cells in a Hormone-Restricted Microenvironment.

The fibril orientation of type I collagen has been shown to contribute to tumor invasion and metabolic changes. Yet, there is limited information about its impact on tumor cells' behavior in a restrictive growth environment. Restrictive growth environments are generated by the inhibition of a proliferation stimulus during therapy or as an inflammatory response to suppress tumor expansion. In this study, the impact of a type I collagen matrix orientation and fibrous architecture on cell proliferation and response to estrogen receptor (ER) therapy were examined using estrogen-dependent breast tumor cells (MCF-7 and T-47D) cultured in a hormone-restricted environment. The use of hormone-free culture media, as well as pharmacological inhibitors of ER, Tamoxifen, and Fulvestrant, were investigated as hormone restrictive conditions. Examination of cultures at 72 h showed that tumor cell proliferation was significantly stimulated (1.8-fold) in the absence of hormones on collagen fibrous substrates, but not on polycaprolactone fibrous substrates of equivalent orientation. ER inhibitors did not suppress cell proliferation on collagen fibrous substrates. The examination of reporter cells for ER signaling showed a lack of activity, thus confirming a shift toward an ER-independent proliferation mechanism. Examination of two selective inhibitors of α2ß1 and α1ß1 integrins showed that cell proliferation is suppressed in the presence of the α2ß1 integrin inhibitor only, thereby indicating that the observed changes in tumor cell behavior are caused by a combination of integrin signaling and/or an intrinsic structural motif that is uniquely present in the collagen fibrils. Adjacent coculture studies on collagen substrates showed that tumor cells on collagen can stimulate the proliferation of cells on tissue culture plastic through soluble factors. The magnitude of this effect correlated with the increased surface anisotropy of the substrate. This sensing in fibril orientation was further supported by a differential expression pattern of secreted proteins that were identified on random and aligned orientation substrates. Overall, this study shows a new role for electrospun collagen I fibrous substrates by supporting a shift toward an ER-independent tumor cell proliferation mechanism in ER+ breast tumor cells.

Protective Action of Jania rubens Nanoencapsulated Algal Extract in Controlling Vegetable Oils' Rancidity.

The development of natural antioxidants that can mitigate oil oxidation is on the rise. Several antioxidants have been developed from natural terrestrial plants, with less emphasis on marine algae. Rancidity is a major degradative reaction limiting the shelf-life and deteriorating the quality of vegetable oils. The goal of this study was to evaluate the ability of the Jania rubens' (J. rubens) seaweed extract encapsulated by chitosan/tripolyphosphate in retarding lipids' oxidation in vegetable oils. To improve the J. rubens efficacy, the extract was nanoencapsulated using the ionic gelation method. A Box-Behnken design was applied for the optimization of the formulation variables (chitosan/tripolyphosphate amounts, homogenization time, and homogenization speed). The optimum nanoformulation was characterized by transmission electron microscopy. It had a particle size of 161 nm, zeta potential of 31.2 mV, polydispersity index of 0.211, and entrapment efficiency of 99.7%. The ability of the optimum formula to extend the shelf-life of vegetable oils was based on peroxide value and thiobarbituric acid assays. In addition, headspace solid-phase microextraction was applied to detect the oils' volatiles as secondary markers of rancidity. The results revealed that the nanoencapsulated algal extract considerably reduced the rate of oils' oxidation and that its activity was comparable to that of a widely used synthetic antioxidant.

Spatiotemporal in vivo tracking of polyclonal human regulatory T cells (Tregs) reveals a role for innate immune cells in Treg transplant recruitment.

Regulatory T cells (Tregs) are emerging as a new cell-based therapy in solid organ transplantation. Adoptive transfer of Tregs has been shown preclinically to protect from graft rejection, and the safety of Treg therapy has been demonstrated in clinical trials. Despite these successes, the in vivo distribution and persistence of adoptively transferred Tregs remained elusive, which hampers clinical translation. Here we isolated human Tregs using a GMP-compatible protocol and lentivirally transduced them with the human sodium iodide symporter to render them traceable in vivo by radionuclide imaging. Engineered human Tregs were characterized for phenotype, survival, suppressive capacity, and reporter function. To study their trafficking behavior, they were subsequently administered to humanized mice with human skin transplants. Traceable Tregs were quantified in skin grafts by non-invasive nano-single-photon emission computed tomography (nanoSPECT)/computed tomography (CT) for up to 40 days, and the results were validated ex vivo. Using this approach, we demonstrated that Treg trafficking to skin grafts was regulated by the presence of recipient Gr-1+ innate immune cells. We demonstrated the utility of radionuclide reporter gene-afforded quantitative Treg in vivo tracking, addressing a fundamental need in Treg therapy development and offering a clinically compatible methodology for future Treg therapy imaging in humans.

The Future of Regulatory T Cell Therapy: Promises and Challenges of Implementing CAR Technology.

Cell therapy with polyclonal regulatory T cells (Tregs) has been translated into the clinic and is currently being tested in transplant recipients and patients suffering from autoimmune diseases. Moreover, building on animal models, it has been widely reported that antigen-specific Tregs are functionally superior to polyclonal Tregs. Among various options to confer target specificity to Tregs, genetic engineering is a particularly timely one as has been demonstrated in the treatment of hematological malignancies where it is in routine clinical use. Genetic engineering can be exploited to express chimeric antigen receptors (CAR) in Tregs, and this has been successfully demonstrated to be robust in preclinical studies across various animal disease models. However, there are several caveats and a number of strategies should be considered to further improve on targeting, efficacy and to understand the in vivo distribution and fate of CAR-Tregs. Here, we review the differing approaches to confer antigen specificity to Tregs with emphasis on CAR-Tregs. This includes an overview and discussion of the various approaches to improve CAR-Treg specificity and therapeutic efficacy as well as addressing potential safety concerns. We also discuss different imaging approaches to understand the in vivo biodistribution of administered Tregs. Preclinical research as well as suitability of methodologies for clinical translation are discussed.

Microfluidic lumen-based systems for advancing tubular organ modeling.

Microfluidic lumen-based systems are microscale models that recapitulate the anatomy and physiology of tubular organs. These technologies can mimic human pathophysiology and predict drug response, having profound implications for drug discovery and development. Herein, we review progress in the development of microfluidic lumen-based models from the 2000s to the present. The core of the review discusses models for mimicking blood vessels, the respiratory tract, the gastrointestinal tract, renal tubules, and liver sinusoids, and their application to modeling organ-specific diseases. We also highlight emerging application areas, such as the lymphatic system, and close the review discussing potential future directions.

Oral intramuscular hemangioma: Report of three cases.

Intramuscular hemangioma (IMH) represents less than 1% of all hemangiomas. In the head and neck region, it occurs mostly in the masseter, temporalis and sternocleidomastoid muscles. Despite its infiltrative growth pattern and several worrisome histological features, such as increased mitotic activity, plumpness of the nuclei, intraluminal papillary projections or perineural infiltration, the lesion is benign, and complete surgical excision is the preferred treatment for such oral lesions. Herein, we report three rare cases of IMH in the tongue and lip, discuss the clinical and histological aspects, and review the literature regarding this lesion.

Open multi-culture platform for simple and flexible study of multi-cell type interactions.

The study of multi-cell-type (MCT) interactions has the potential to significantly impact our understanding of tissue and disease biology. Such studies require innovative culture tools for unraveling the contributions of each cell type. Micro- and macro-scale platforms for MCT culture each have different advantages and disadvantages owing to their widely different capabilities, availability, and ease-of-use. However, as evidenced in the literature, there are very few examples of MCT studies and culture platforms, suggesting both biological and technical barriers. We have developed an open multi-culture platform to promote more rapid progress by integrating advantages of both micro- and macro-scale culture devices. The proposed open multi-culture platform addresses technical barriers by allowing easy customization, independent control of basic physical culture parameters, and incorporation of multiple culture modalities (e.g., 2D, 3D, transwell, and spheroid). The design also permits the user to obtain independent endpoints for each culture region. We demonstrate use of the platform in two example studies where we evaluated how cell ratio and cell types influence the response of triple negative breast cancer cells to heat damage and Hedgehog signaling. We also show that the platform can improve soluble factor transport between cell types compared to compartmentalized macro- and micro-scale alternatives. Last, we examine current and future challenges of the platform. We envision simple, yet flexible and customizable, platforms such as this will be important for advancing in vitro study of tissue and tumor biology.

Razor-printed sticker microdevices for cell-based applications.

Tape-based razor-printing is a flexible and affordable ultra-rapid prototyping approach for microscale device fabrication. However, integration of this prototyping approach into cell-based assay development has been limited to proof of principle demonstrations. This is in large part due to lack of an established or well-characterized option for biocompatible adhesive tape. Without such an option, integration of these areas will remain unexplored. Therefore, to address this critical hurdle, we characterized microscale devices made using a potentially biocompatible double-sided adhesive, ARCare 90106. We validated tape-based device performance against 96-well plates and PDMS microdevices with respect to cell viability, hydrophobic small molecule sequestration, the potential for leaching compounds, use in fluorescence microscopy, and outgassing (bubble formation). Results supported the tape as a promising tool for future cell-based assay development. Therefore, we subsequently demonstrated specific strengths enabled by the ultra-rapid (<1 h per prototype) and affordable (∼$1200 cutting plotter, <$0.05 per prototype) approach. Specifically, data demonstrate the ability to integrate disparate materials for advanced sticker-device functionality such as bonding of polystyrene devices to glass substrates for microscopy applications, inclusion of membranes, and incorporation of different electrospun biomaterials into a single device. Likewise, the approach allowed rapid adoption by uninitiated users. Overall, this study provides a necessary and unique contribution to the largely separate fields of tape-based razor-printing and cell-based microscale assay development by addressing a critical barrier to widespread integration and adoption while also demonstrating the potential for new and future applications.

Microbial volatile communication in human organotypic lung models.

We inhale respiratory pathogens continuously, and the subsequent signaling events between host and microbe are complex, ultimately resulting in clearance of the microbe, stable colonization of the host, or active disease. Traditional in vitro methods are ill-equipped to study these critical events in the context of the lung microenvironment. Here we introduce a microscale organotypic model of the human bronchiole for studying pulmonary infection. By leveraging microscale techniques, the model is designed to approximate the structure of the human bronchiole, containing airway, vascular, and extracellular matrix compartments. To complement direct infection of the organotypic bronchiole, we present a clickable extension that facilitates volatile compound communication between microbial populations and the host model. Using Aspergillus fumigatus, a respiratory pathogen, we characterize the inflammatory response of the organotypic bronchiole to infection. Finally, we demonstrate multikingdom, volatile-mediated communication between the organotypic bronchiole and cultures of Aspergillus fumigatus and Pseudomonas aeruginosa.

A Comparison of 2 Respiratory Devices for Sputum Clearance in Adults With Non-Cystic Fibrosis Bronchiectasis.

BACKGROUND: Airway clearance techniques are a vital part of routine care for patients with bronchiectasis. There is no clear superior modality. The Flutter combines oscillations (6-20 Hz) and positive expiratory pressure; the Lung Flute combines positive expiratory pressure and low frequency acoustic waves (18-22 Hz), to augment clearance. This project aimed to compare these devices. METHODS: This was a randomized crossover study of adult subjects with stable non-cystic fibrosis bronchiectasis (expectorating > 25 mL/d). Subjects attended 2 separate out-patient visits, 1 week apart, and completed a supervised sputum clearance regime and Lickert scale (8 questions regarding subjects' perception of the experience using each device). Total sputum expectorated during supervised intervention (T1) and after 30 min from the end of T1 (T2) was recorded as wet sputum weight. Total wet sputum weight desiccated in a microwave (10 min at 300 watts), allowed measurement of total dry sputum weight. Data were compared using paired t test. RESULTS: We recruited 40 subjects with a mean ± SD age of 63 ± 16 y. Overall, there was no significant difference in wet sputum weight (Flutter, 5.78 ± 6.47 g; Lung Flute, 5.75 ± 0.22 g) and dry sputum weight (Flutter, 0.40 ± 0.86 g; Lung Flute, 0.22 ± 0.21 g). At T1, wet sputum weight was higher for the Flutter (5.10 ± 6.26 g) compared with the Lung Flute (3.74 ± 3.44 g) (P = .038). At T2, wet sputum weight was higher for the Lung Flute (2.02 ± 3.01 g) compared with the Flutter (0.68 ± 0.75 g) (P = .001). Subjects perceived the Flutter as being significantly better at clearing secretions (P = .01), easy to understand (P = .03), and simple to use (P = .01) compared with the Lung Flute. CONCLUSIONS: Both devices were well-tolerated and successfully augmented secretion clearance. Most subjects preferred the Flutter because of increased speed of secretion clearance, and greater ease of use.

Physical activity levels in Bangladeshi adults: results from STEPS survey 2010.

OBJECTIVES: Physical inactivity is an established risk factor for non-communicable diseases (NCD) and identified as the major public health concern worldwide. However, nationally representative and internationally comparable data on physical activity (PA) are lacking in Bangladesh. The objective of this paper was to determine nationally representative prevalence of PA levels among Bangladeshi adults. STUDY DESIGN: Cross-sectional survey. METHODS: Data, on PA for this paper, were analysed from the NCD risk factors survey 2010 in Bangladesh. A standardized approach known as STEPS (STEPSwise approach to Surveillance for NCD risk factors) was followed for this survey. A total of 9275 adults (aged ≥ 25 years) were interviewed. Data on PA were processed and analysed according to Global Physical Activity Questionnaire (GPAQ) version 2 analysis framework. RESULTS: Of total 9275 respondents 4312 were men and 4963 women with a mean age of 42.4 (±13.5) years. Median MET-minutes of total PA in a typical week was double in rural areas (3360) than urban (1680) areas. The overall country wide prevalence of low PA was 34.5% (95% confidence interval, 33.5-35.5), urban 37.7% (36.3-39.1) and rural 31.6% (30.3-32.9). Women in general were more inactive (women, 53.6% [52.2-55.0], men 15.4% [14.9-17.1]). The main contributions to total PA were from work (urban 47.0%, rural 61.0%), and active commuting (38.0%, 30.0%) domains. Leisure-time PA represented only a small proportion (15.0%, 9.0%). CONCLUSIONS: Insufficient physical activity is highly prevalent among the Bangladeshi adult population. Promoting overall PA at leisure-time and commuting considering country context can be feasible options with special attention to the women.