Implantation of short-term biventricular assist device (BiVAD) using the CentriMag™ system: the Manchester technique.

Biventricular assist devices (BiVADs) using the CentriMag™ system are being used increasingly as a form of short-term mechanical circulatory support for the treatment of acute cardiogenic shock from any aetiology. They can be used as a bridge to decision, recovery or transplantation. BiVADs are associated with better clinical outcomes when compared to veno-arterial (VA) extracorporeal membrane oxygenator (ECMO) systems. In this paper, we describe a safe and reproducible method of BiVAD implantation using the CentriMag™ system at our institution.

Tricks and tracks of prevalence, occurrences, treatment technologies, and challenges of mixtures of emerging contaminants in the environment: With special emphasis on microplastic.

This paper aims to emphasize the occurrence of various emerging contaminant (EC) mixtures in natural ecosystems and highlights the primary concern arising from the unregulated release into soil and water, along with their impacts on human health. Emerging contaminant mixtures, including pharmaceuticals, personal care products, dioxins, polychlorinated biphenyls, pesticides, antibiotics, biocides, surfactants, phthalates, enteric viruses, and microplastics (MPs), are considered toxic contaminants with grave implications. MPs play a crucial role in transporting pollutants to aquatic and terrestrial ecosystems as they interact with the various components of the soil and water environments. This review summarizes that major emerging contaminants (ECs), like trimethoprim, diclofenac, sulfamethoxazole, and 17α-Ethinylestradiol, pose serious threats to public health and contribute to antimicrobial resistance. In addressing human health concerns and remediation techniques, this review critically evaluates conventional methods for removing ECs from complex matrices. The diverse physiochemical properties of surrounding environments facilitate the partitioning of ECs into sediments and other organic phases, resulting in carcinogenic, teratogenic, and estrogenic effects through active catalytic interactions and mechanisms mediated by aryl hydrocarbon receptors. The proactive toxicity of ECs mixture complexation and, in part, the yet-to-be-identified environmental mixtures of ECs represent a blind spot in current literature, necessitating conceptual frameworks for assessing the toxicity and risks with individual components and mixtures. Lastly, this review concludes with an in-depth exploration of future scopes, knowledge gaps, and challenges, emphasizing the need for a concerted effort in managing ECs and other organic pollutants.

Dealing With Pericylindrical Melts in Keratoprosthesis: Tenon Patch Graft to the Rescue.

PURPOSE: The aim of this study was to describe the outcomes of autologous Tenon patch graft in the management of Auro keratoprosthesis-related pericylindrical corneal melt. METHODS: We report 3 cases of sterile pericylindrical corneal melt in patients with Auro keratoprosthesis implantation after a mean duration of 5 years (1.5-8 years). Case 1 was a patient with severe graft-versus-host disease. Cases 2 and 3 were cases of chemical injury.All these cases of sterile pericylindrical corneal melt (4-6 mm) underwent autologous Tenon patch graft. The technique included freshening of the edges around the melt, followed by measuring the size of the defect. A Tenon graft harvested from the patient's own eye was used to seal the defect and act as a scaffold. The Tenon patch graft was spread over the melt and held in place by the application of fibrin glue and/or interrupted 10-0 nylon sutures. A bandage contact lens was then placed on the eye. RESULTS: Tenon patch graft was well taken in all patients. The mean duration of epithelial healing was 1 month. Globe integrity was well maintained with no postoperative complications at a mean follow-up duration of 12 months (6-18 months). CONCLUSIONS: Corneal melt is one of the most dreaded complications of KPro because its occurrence could threaten visual prognosis and globe integrity. Autologous Tenon patch is a simple yet innovative and effective option to steer such eyes away from potentially dreadful complications.

Managing construction and demolition waste using lean tools to achieve environmental sustainability: an Indian perspective.

The construction industry substantially impacts a country's economic growth and ecological progress. Due to the competitive nature of the construction business and intense rivalry between construction companies, the industry's focus is progressively becoming customer-centric. Efforts are being taken to ensure high-quality buildings are built at reasonable rates and on time. Companies are trying to ensure on-time building projects are completed within allocated budgets. This can be accomplished by ensuring minimal waste generation from various activities during the construction process. Implementing lean tools and techniques can improve work efficiency and reduce waste in the building process. This research study offers a basic understanding of lean tools and emphasizes their contribution in terms of time, effort, and sustainability. The primary purpose of this study is to present the effectiveness of the lean process for construction waste management. Through a case study, this research shows the deployment of lean tools and principles for efficient construction waste management and optimal use of building resources. The improvement in reduced waste generation and enhanced organizational resource productivity is closely monitored. The study results indicated that the lean framework could reduce waste by 25 to 50%. It is demonstrated that lean construction significantly improves construction sustainability and productivity. If it is implemented along with automation tools and circular economy concepts, more than 50% of waste reduction can be achieved. These viable initiatives are necessary to improve the performance of the Indian construction industry to achieve circularity in waste management which indirectly helps in the sustainability goals of the construction sector.

Secondary intrachoroidal cavitation in a case of iridofundal coloboma.

Intrachoroidal cavitation is a morphological entity usually described in high myopic eyes. It is usually seen in peripapillary location, though macular location has also been reported. It has been associated with optic disc tilting and posterior staphyloma. We report a case of a 27 year old female who presented to us with recent onset scotoma in her visual field. On examination she had a fundal coloboma with intercalary membrane detachment with intrachoroidal cavitation at the edge of the coloboma. We hypothetise that this intrachoroidal cavitation is a secondary phenomenon and is responsible for recent onset scotoma in her visual field.

Enhancing mitochondrial function in vivo rescues MDS-like anemia induced by pRb deficiency.

Erythropoiesis is intimately coupled to cell division, and deletion of the cell cycle regulator retinoblastoma protein (pRb) causes anemia in mice. Erythroid-specific deletion of pRb has been found to result in inefficient erythropoiesis because of deregulated coordination of cell cycle exit and mitochondrial biogenesis. However, the pathophysiology remains to be fully described, and further characterization of the link between cell cycle regulation and mitochondrial function is needed. To this end we further assessed conditional erythroid-specific deletion of pRb. This resulted in macrocytic anemia, despite elevated levels of erythropoietin (Epo), and an accumulation of erythroid progenitors in the bone marrow, a phenotype strongly resembling refractory anemia associated with myelodysplastic syndromes (MDS). Using high-fractionation fluorescence-activated cell sorting analysis for improved phenotypic characterization, we illustrate that erythroid differentiation was disrupted at the orthochromatic stage. Transcriptional profiling of sequential purified populations revealed failure to upregulate genes critical for mitochondrial function such as Pgc1ß, Alas2, and Abcb7 specifically at the block, together with disturbed heme production and iron transport. Notably, deregulated ABCB7 causes ring sideroblastic anemia in MDS patients, and the mitochondrial co-activator PGC1ß is heterozygously lost in del5q MDS. Importantly, the anemia could be rescued through enhanced PPAR signaling in vivo via either overexpression of Pgc1ß or bezafibrate administration. In conclusion, lack of pRb results in MDS-like anemia with disrupted differentiation and impaired mitochondrial function at the orthochromatic erythroblast stage. Our findings reveal for the first time a role for pRb in heme and iron regulation, and indicate that pRb-induced anemia can be rescued in vivo through therapeutic enhancement of PPAR signaling.

Lysine-specific demethylase 1A restricts ex vivo propagation of human HSCs and is a target of UM171.

Culture conditions in which hematopoietic stem cells (HSCs) can be expanded for clinical benefit are highly sought after. Here, we report that inhibition of the epigenetic regulator lysine-specific histone demethylase 1A (LSD1) induces a rapid expansion of human cord blood-derived CD34+ cells and promotes in vitro propagation of long-term repopulating HSCs by preventing differentiation. The phenotype and molecular characteristics of cells treated with LSD1 inhibitors were highly similar to cells treated with UM171, an agent promoting expansion of HSCs through undefined mechanisms and currently being tested in clinical trials. Strikingly, we found that LSD1, as well as other members of the LSD1-containing chromatin remodeling complex CoREST, is rapidly polyubiquitinated and degraded upon UM171 treatment. CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 depletion of the CoREST core member, RCOR1, resulted in expansion of CD34+ cells similar to LSD1 inhibition and UM171. Taken together, LSD1 and CoREST restrict HSC expansion and are principal targets of UM171, forming a mechanistic basis for the HSC-promoting activity of UM171.

Biogenic stabilization and heavy metal immobilization during vermicomposting of vegetable waste with biochar amendment.

Vermicomposting is a traditional technology that produces the best quality of compost, but factors such as maturity, presence of heavy metals, etc. need to be tackled prior to agrarian application. The present study investigates the influence of varying biochar dose (2.5, 5, and 10% on a weight basis) on the maturity of compost and heavy metals during vermicomposting of vegetable waste using epigeic earthworm. Biochar amendment notably enhanced the electrical conductivity (up to 2.7 mS/cm), nitrogen content (up to 3.1%), NO3-N (up to 630 mg/kg) and nutritional value. The heavy metals, oxygen uptake rate (below 0.96 mg/g VS/day) and CO2 evolution rate (below 1 mg/g VS/day) were attenuated along with degradation of complex organic crystals as observed in powder X-Ray Diffraction (PXRD) spectra. Furthermore, biochar aid in reducing pathogens (below 1.1 × 103 MPN/g dry weight) as inferred from the Most Probable Number (MPN) results as well as degrading the complex organics into simpler compounds as revealed from the Fourier-transform infrared spectroscopy (FTIR) spectra. The present study inferred that the vegetable waste was biologically stabilized through biochar amendment during vermicomposting process with improved nutritional and physico-chemical properties.

Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids.

One of the hallmarks of cancers is their ability to develop resistance against therapeutic agents. Therefore, developing effective in vitro strategies to identify drug resistance remains of paramount importance for successful treatment. One of the ways cancer cells achieve drug resistance is through the expression of efflux pumps that actively pump drugs out of the cells. To date, several studies have investigated the potential of using 3-dimensional (3D) multicellular tumor spheroids (MCSs) to assess drug resistance; however, a unified system that uses MCSs to differentiate between multi drug resistance (MDR) and non-MDR cells does not yet exist. In the present report we describe MCSs obtained from post-diagnosed, pre-treated patient-derived (PTPD) cell lines from head and neck squamous cancer cells (HNSCC) that often develop resistance to therapy. We employed an integrated approach combining response to clinical drugs and screening cytotoxicity, monitoring real-time drug uptake, and assessing transporter activity using flow cytometry in the presence and absence of their respective specific inhibitors. The report shows a comparative response to MDR, drug efflux capability and reactive oxygen species (ROS) activity to assess the resistance profile of PTPD MCSs and two-dimensional (2D) monolayer cultures of the same set of cell lines. We show that MCSs provide a robust and reliable in vitro model to evaluate clinical relevance. Our proposed strategy can also be clinically applicable for profiling drug resistance in cancers with unknown resistance profiles, which consequently can indicate benefit from downstream therapy.

Recalcitrant carbon for composting of fibrous aquatic waste: Degradation kinetics, spectroscopic study and effect on physico-chemical and nutritional properties.

Biochar, a recalcitrant carbon, is known to enhance organic matter degradation and improve physical properties. The objective of the study is to examine the probable effect of biochar addition during composting of a fibrous aquatic waste, i.e., water hyacinth though degradation kinetics and spectroscopic (FTIR and PXRD) analysis. Four dosages of biochar (0, 2.5, 5, and 10% w/w) were mixed to a mixture of water hyacinth, cow-dung and saw-dust comprising a total weight of 150 kg and composted using rotary drum composter for 20 days in batch mode. The study outcomes indicated that the amendment of biochar prolonged the duration of the thermophilic temperatures, reduced salinity, and promoted nutritional quality of compost. Moreover, biochar amendment enhanced the organic matter degradation with a rate constant of 0.029 day-1 and increased the total Kjeldahl nitrogen content up to 1.75% from an initial value of 1.10% in the reactor with 2.5% biochar amendment. Concurrently, biochar amendment aided in reducing Cu and Cr in the final product inferring 2.5% biochar is best suited for composting of water hyacinth. However, future studies are encouraged to decipher the microbial shifts and bioavailability of metals due to biochar dosage during composting for mitigating and managing the menace of such fibrous waste like water hyacinth by converting it to a soil conditioner.

Interplay of physical and chemical properties during in-vessel degradation of sewage sludge.

Sewage sludge produced is either applied to land or used as fertilizer for crops or disposed of in landfills, causing several environmental problems. Recent studies revealed that composting is a proven technology in reducing organic content, heavy metals, and harmful pathogens, improving the nutritional value of sewage sludge, which is useful for crops. But studies on variation in physical properties are rare. Composting physics or physical properties during composting plays a vital role from handling, management, and utilization of end product, i.e., compost. This study mainly deals with the detailed information on physics involved during the degradation process, which is crucial for land and geotechnical applications. In the present study, sewage sludge was used as a composting substrate in 550 L in-vessel rotary drum composter. Emphasis was given in deciphering the changes in physical parameters such as bulk density, porosity, and air-filled porosity and few chemical parameters during the composting process. Besides, a relationship between different physical properties during rotary drum composting was investigated statistically. Bulk density was observed to have increased from 643 to 707 kg m-3 as a result of volume reduction of compost matrix. Moreover, the gravimetric moisture content was found to be less than 45% in the end product, which is recommended for compost.

The effect of 2D and 3D cell cultures on treatment response, EMT profile and stem cell features in head and neck cancer.

BACKGROUND: Head and Neck Squamous Cell Carcinoma (HNSCC) tumors are often resistant to therapies. Therefore searching for predictive markers and new targets for treatment in clinically relevant in vitro tumor models is essential. Five HNSCC-derived cell lines were used to assess the effect of 3D culturing compared to 2D monolayers in terms of cell proliferation, response to anti-cancer therapy as well as expression of EMT and CSC genes. METHODS: The viability and proliferation capacity of HNSCC cells as well as induction of apoptosis in tumor spheroids cells after treatment was assessed by MTT assay, crystal violet- and TUNEL assay respectively. Expression of EMT and CSC markers was analyzed on mRNA (RT-qPCR) and protein (Western blot) level. RESULTS: We showed that HNSCC cells from different tumors formed spheroids that differed in size and density in regard to EMT-associated protein expression and culturing time. In all spheroids, an up regulation of CDH1, NANOG and SOX2 was observed in comparison to 2D but changes in the expression of EGFR and EMT markers varied among the cell lines. Moreover, most HNSCC cells grown in 3D showed decreased sensitivity to cisplatin and cetuximab (anti-EGFR) treatment. CONCLUSIONS: Taken together, our study points at notable differences between these two cellular systems in terms of EMT-associated gene expression profile and drug response. As the 3D cell cultures imitate the in vivo behaviour of neoplastic cells within the tumor, our study suggest that 3D culture model is superior to 2D monolayers in the search for new therapeutic targets.

Drum composting of nitrogen-rich Hydrilla Verticillata with carbon-rich agents: Effects on composting physics and kinetics.

Composting of the Hydrilla verticillata, an invasive aquatic weed, signifies aquatic waste management as a safe and hygienic method that produces a nutrient-rich end product, i.e., compost. However, its higher moisture content, higher N-losses, and lower degradation rate have shown negative impacts on the composting process. Therefore the primary objective of this study was to assess the composting physics and the degradation kinetics after addition of three different carbon-rich agents with Hydrilla verticillata. To pursue this objective, three carbon-rich agents (viz. dry leaves in Run A, grass clippings in Run B and wood chips in Run C) each were mixed (10% w/w) to the optimized control mixture of Hydrilla verticillata, cow dung and sawdust (8:1:1) as reported in the earlier study. The composting experiments were performed in 550L rotary drum composter for 20 days to evaluate variation in physical, chemical, nutritional properties as well as degradation kinetics. The Run A and Run B were the only two mixtures that attained the temperature (55-70 °C) that indicates standard sterilization capacity in both with maximum moisture reduction (17%) and total Kjeldahl N increment (48%) in the latter. Organic matter losses throughout the process followed a first-order kinetic equation in all the Run (A-C) and control with the higher loss in Run B whereas least in control. Nevertheless, the addition of all carbon-rich agents is found to be beneficial to improve composting physics. Amongst all Runs (A-C), Run B achieved maximum reduction in the initial value of bulk density (64%) and increment in the initial value of free air space (20%). The study also concluded that all the carbon-rich agents have produced compost with the nutritional concentration suitable for agricultural proposes.

Efficacy of batch mode rotary drum composter for management of aquatic weed (Hydrilla verticillata (L.f.) Royle).

Invasive aquatic weed management is one of the biggest challenges in the field of solid waste management. Eichhornia crassipes, Pistia stratiotes and Hydrilla verticillata (L.f.) Royle pose some of the world's most noted aquatic weed problems. Previously reported studies on management of H. verticillata, a submersed aquatic plant, have shown that temporary removal, chemical treatment or biological control methods each have advantages and disadvantages. Removal programs that propose to compost harvested H. verticillata biomass may provide a novel technique to manage this issue. However, the properties of such compost as an agriculture resource are unclear. This study presents the different mix proportions of H. verticillata, cow dung and sawdust used for the composting of 550 L rotary drum composter. This work characterizes the biological, physicochemical, and respirometry properties of the various mixes over a 20-day composting period. The results suggest that the biomass of H. verticillata can be beneficially utilized to produce stable compost for potential use in agricultural systems.

Biochar amendment for batch composting of nitrogen rich organic waste: Effect on degradation kinetics, composting physics and nutritional properties.

Composting is an efficient technology to reduce pathogenic bodies and stabilize the organic matter in organic wastes. This research work investigates an effect of biochar as amendment to improve the composting efficiency and its effect on degradation kinetics, physical and nutritional properties. Biochar (2.5, 5 and 10% (w/w)) were added into a mixture of Hydrilla verticillata, cow dung and sawdust having ratio of 8:1:1 (control), respectively. Biochar addition resulted in advanced thermophilic temperatures (59 °C) and could improve the physical properties of composting process. Owing to addition of 5% biochar as a bulking agent in composting mixture, the final product from composting, total nitrogen increased by 45% compared to the other trials, and air-filled porosity decreased by 39% and was found to be within recommended range from literature studies. Considering temperature, degradation rate and nitrogen transformation the amendment of 5% biochar is recommended for Hydrilla verticillata composting.

In Vitro Tumorigenic Assay: Colony Forming Assay for Cancer Stem Cells.

Colony forming or clonogenic assay is an in vitro quantitative technique to examine the capability of a single cell to grow into a large colony through clonal expansion. Clonogenic activity is a sensitive indicator of undifferentiated cancer stem cells. Here, we described the colony forming ability of the isolated breast cancer stem cells from the total population of cancer cells using double-layered, soft agarose-based assay. This method demonstrates that cancer stem cells can survive and generate colony growth in an anchorage-independent culture model. The 0.005% crystal violet solution is used in this assay to visualize the generated colonies.

Inhibition of miR301 enhances Akt-mediated cell proliferation by accumulation of PTEN in nucleus and its effects on cell-cycle regulatory proteins.

Micro-RNAs (miRs) represent an innovative class of genes that act as regulators of gene expression. Recently, the aberrant expression of several miRs has been associated with different types of cancers. In this study, we show that miR301 inhibition influences PI3K-Akt pathway activity. Akt overexpression in MCF7 and MDAMB468 cells caused downregulation of miR301 expression. This effect was confirmed by co-transfection of miR301-modulators in the presence of Akt. Cells overexpressing miR301-inhibitor and Akt, exhibited increased migration and proliferation. Experimental results also confirmed PI3K, PTEN and FoxF2 as regulatory targets for miR301. Furthermore, Akt expression in conjunction with miR301-inhibitor increased nuclear accumulation of PTEN, thus preventing it from downregulating the PI3K-signalling. In summary, our data emphasize the importance of miR301 inhibition on PI3K-Akt pathway-mediated cellular functions. Hence, it opens new avenues for the development of new anti-cancer agents preferentially targeting PI3K-Akt pathway.

The expression pattern of PFKFB3 enzyme distinguishes between induced-pluripotent stem cells and cancer stem cells.

Induced pluripotent stem cells (iPS) have become crucial in medicine and biology. Several studies indicate their phenotypic similarities with cancer stem cells (CSCs) and a propensity to form tumors. Thus it is desirable to identify a trait which differentiates iPS populations and CSCs. Searching for such a feature, in this work we compare the restriction (R) point-governed regulation of cell cycle progression in different cell types (iPS, cancer, CSC and normal cells) based on the expression profile of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase3 (PFKFB3) and phosphofructokinase (PFK1). Our study reveals that PFKFB3 and PFK1 expression allows discrimination between iPS and CSCs. Moreover, cancer and iPS cells, when cultured under hypoxic conditions, alter their expression level of PFKFB3 and PFK1 to resemble those in CSCs. We also observed cell type-related differences in response to inhibition of PFKFB3. This possibility to distinguish CSC from iPS cells or non-stem cancer cells by PFKB3 and PFK1 expression improves the outlook for clinical application of stem cell-based therapies and for more precise detection of CSCs.

Role of the salt bridge between glutamate 546 and arginine 907 in preservation of autoinhibited form of Apaf-1.

Apaf-1, the key element of apoptotic mitochondrial pathway, normally exists in an auto-inhibited form inside the cytosol. WRD-domain of Apaf-1 has a critical role in the preservation of auto-inhibited form; however the underlying mechanism is unclear. It seems the salt bridges between WRD and NOD domains are involved in maintaining the inactive conformation of Apaf-1. At the present study, we have investigated the effect of E546-R907 salt bridge on the maintenance of auto-inhibited form of human Apaf-1. E546 is mutated to glutamine (Q) and arginine (R). Over-expression of wild type Apaf-1 and its E546Q and E546R variants in HEK293T cells does not induce apoptosis unlike - HL-60 cancer cell line. In vitro apoptosome formation assay showed that all variants are cytochrome c and dATP dependent to form apoptosome and activate endogenous procaspase-9 in Apaf-1-knockout MEF cell line. These results suggest that E546 is not a critical residue for preservation of auto-inhibited Apaf-1. Furthermore, the behavior of Apaf-1 variants for in vitro apoptosome formation in HEK293T cell is similar to exogenous wild type Apaf-1. Wild type and its variants can form apoptosome in HEK293T cell with different procaspase-3 processing pattern in the presence and absence of exogenous cytochrome c and dATP.

Nuclear localized Akt enhances breast cancer stem-like cells through counter-regulation of p21(Waf1/Cip1) and p27(kip1).

UNLABELLED: Cancer stem-like cells (CSCs) are a rare subpopulation of cancer cells capable of propagating the disease and causing cancer recurrence. In this study, we found that the cellular localization of PKB/Akt kinase affects the maintenance of CSCs. When Akt tagged with nuclear localization signal (Akt-NLS) was overexpressed in SKBR3 and MDA-MB468 cells, these cells showed a 10-15% increase in the number of cells with CSCs enhanced ALDH activity and demonstrated a CD44(+High)/CD24(-Low) phenotype. This effect was completely reversed in the presence of Akt-specific inhibitor, triciribine. Furthermore, cells overexpressing Akt or Akt-NLS were less likely to be in G0/G1 phase of the cell cycle by inactivating p21(Waf1/Cip1) and exhibited increased clonogenicity and proliferation as assayed by colony-forming assay (mammosphere formation). Thus, our data emphasize the importance the intracellular localization of Akt has on stemness in human breast cancer cells. It also indicates a new robust way for improving the enrichment and culture of CSCs for experimental purposes. Hence, it allows for the development of simpler protocols to study stemness, clonogenic potency, and screening of new chemotherapeutic agents that preferentially target cancer stem cells. SUMMARY: The presented data, (i) shows new, stemness-promoting role of nuclear Akt/PKB kinase, (ii) it underlines the effects of nuclear Akt on cell cycle regulation, and finally (iii) it suggests new ways to study cancer stem-like cells.