Multistep Pathway Research Articles

Kinetics and thermodynamics of non‐isothermal pyrolysis of Terminalia chebula branches at different heating rates

AbstractNon‐isothermal thermogravimetric tests of Terminalia chebula (Helikha) were conducted under inert N2 gas environment for temperatures (25–900°C) at heating rates of 10, 20, 35, and 55°C min−1. Kinetic triplet approximated employing five iso‐conversional methods namely, differential Friedman method (DFM), distributed activation method (DAEM), Ozawa–Flynn–Wall (OFW), Kissinger–Akahira–Sunose (KAS), and Starink (STK). Average values of activation energy (kJ mol−1) and frequency factor (min−1) calculated by the five models were 227.11, 2.98 × 1021 for DFM; 229.21, 4.63 × 1021 for KAS; 227.11, 3.81 × 1020 for OFW; 225.54, 1.15 × 1018 for STK; and 227.33, 3.02 × 1020 for DAEM respectively over the conversion range up to 0.8. In the kinetics study, correlation coefficient (R2) of greater than 0.97 is noticed in the conversion range of α = 0.1–0.8 for all models. From thermodynamic analysis, average values of ΔH (kJ mol−1), ΔG (kJ mol−1), and ΔS (kJ mol−1 K−1) for DAEM: 221.8, 179.69, and 0.065; for DFM: 236.40, 179.37, and 0.089; for KAS: 221.8, 179.69, and 0.065; for OFW: 220.22, 179.72, and 0.063; and for STK: 222.02, 179.68, and 0.066 were estimated to assess viability and reactivity of the process. Criado's master plots revealed that the data obtained from pyrolysis of selected biomass was followed a multistep reaction pathway.

Efficient myrcene production using linalool dehydratase isomerase and rational biochemical process in Escherichia coli.

Microbial synthesis of plant-based myrcene is of great interest because of its high demand, however, achieving high biosynthetic titers remains a great challenge. Previous strategies adopted for microbial myrcene production have relied on the recruitment of a multi-step biosynthetic pathway which requires complex metabolic regulation or high activity of myrcene synthase, hindering its application. Here, we present an effective one-step biotransformation system for myrcene biosynthesis from geraniol, using a linalool dehydratase isomerase (LDI) to overcome these limitations. The truncated LDI possesses nominal activity that catalyzes the isomerization of geraniol to linalool and the subsequent dehydration to myrcene in anaerobic environment. In order to improve the robustness of engineered strains for the efficient conversion of geraniol to myrcene, rational enzyme modification and a series of biochemical process engineering were employed to maintain and improve the anaerobic catalytic activity of LDI. Finally, by introducing the optimized myrcene biosynthetic capability in the existing geraniol-production strain, we achieve de novo biosynthesis of myrcene at 1.25g/L from glycerol during 84h aerobic-anaerobic two-stage fermentation, which is much higher than previously reported myrcene levels. This work highlights the value of dehydratase isomerase-based biocatalytic in establishing novel biosynthetic pathways and lays a reliable foundation for the microbial synthesis of myrcene.

A novel CRISPR/Cas9 system with high genomic editing efficiency and recyclable auxotrophic selective marker for multiple-step metabolic rewriting in Pichia pastoris

The methylotrophic budding yeast Pichia pastoris has been utilized to the production of a variety of heterologous recombinant proteins owing to the strong inducible alcohol oxidase promoter (pAOX1). However, it is difficult to use P. pastoris as the chassis cell factory for high-valuable metabolite biosynthesis due to the low homologous recombination (HR) efficiency and the limitation of handy selective markers, especially in the condition of multistep biosynthetic pathways. Hence, we developed a novel CRISPR/Cas9 system with highly editing efficiencies and recyclable auxotrophic selective marker (HiEE-ReSM) to facilitate cell factory in P. pastoris. Firstly, we improved the HR rates of P. pastoris through knocking out the non-homologous-end-joining gene (Δku70) and overexpressing HR-related proteins (RAD52 and RAD59), resulting in higher positive rate compared to the basal strain, achieved 97%. Then, we used the uracil biosynthetic genes PpURA3 as the reverse screening marker, which can improve the recycling efficiency of marker. Meanwhile, the HR rate is still 100% in uracil auxotrophic yeast. Specially, we improved the growth rate of uracil auxotrophic yeast strains by overexpressing the uracil transporter (scFUR4) to increase the uptake of exogenous uracil from medium. Meanwhile, we explored the optimal concentration of uracil (90 mg/L) for strain growth. In the end, the HiEE-ReSM system has been applied for the inositol production (250 mg/L) derived from methanol in P. pastoris. The systems will contribute to P. pastoris as an attractive cell factory for the complex compound biosynthesis through multistep metabolic pathway engineering and will be a useful tool to improve one carbon (C1) bio-utilization.

Experimental and Computational Mechanisms that Govern Long-Term Stability of CO2-Adsorbed ZIF-8-Based Porous Liquids.

Porous liquids (PLs) based on the zeolitic imidazole framework ZIF-8 are attractive systems for carbon capture since the hydrophobic ZIF framework can be solvated in aqueous solvent systems without porous host degradation. However, solid ZIF-8 is known to degrade when exposed to CO2 in wet environments, and therefore the long-term stability of ZIF-8-based PLs is unknown. Through aging experiments, the long-term stability of a ZIF-8 PL formed using the water, ethylene glycol, and 2-methylimidazole solvent system was systematically examined, and the mechanisms of degradation were elucidated. The PL was found to be stable for several weeks, with no ZIF framework degradation observed after aging in N2 or air. However, for PLs aged in a CO2 atmosphere, formation of a secondary phase occurred within 1 day from the degradation of the ZIF-8 framework. From the computational and structural evaluation of the effects of CO2 on the PL solvent mixture, it was identified that the basic environment of the PL caused ethylene glycol to react with CO2 forming carbonate species. These carbonate species further react within the PL to degrade ZIF-8. The mechanisms governing this process involves a multistep pathway for PL degradation and lays out a long-term evaluation strategy of PLs for carbon capture. Additionally, it clearly demonstrates the need to examine the reactivity and aging properties of all components in these complex PL systems in order to fully assess their stabilities and lifetimes.

The Potential Role of Epigallocatechin-3-Gallate (EGCG) in Breast Cancer Treatment

Breast cancer is one of the most diagnosed cancers worldwide, with an incidence of 47.8%. Its treatment includes surgery, radiotherapy, chemotherapy, and antibodies giving a mortality of 13.6%. Breast tumor development is driven by a variety of signaling pathways with high heterogeneity of surface receptors, which makes treatment difficult. Epigallocatechin-3-gallate (EGCG) is a natural polyphenol isolated as the main component in green tea; it has shown multiple beneficial effects in breast cancer, controlling proliferation, invasion, apoptosis, inflammation, and demethylation of DNA. These properties were proved in vitro and in vivo together with synergistic effects in combination with traditional chemotherapy, increasing the effectiveness of the treatment. This review focuses on the effects of EGCG on the functional capabilities acquired by breast tumor cells during its multistep development, the molecular and signal pathways involved, the synergistic effects in combination with current drugs, and how nanomaterials can improve its bioavailability on breast cancer treatment.

Molecular Simulations of Conformational Transitions within the Insulin Receptor Kinase Reveal Consensus Features in a Multistep Activation Pathway.

Modulating the transitions between active and inactive conformations of protein kinases is the primary means of regulating their catalytic activity, achieved by phosphorylation of the activation loop (A-loop). To elucidate the mechanism of this conformational activation, we applied the string method to determine the conformational transition path of insulin receptor kinase between the active and inactive conformations and the corresponding free-energy profiles with and without A-loop phosphorylation. The conformational change was found to proceed in three sequential steps: first, the flipping of the DFG motif of the active site; second, rotation of the A-loop; finally, the inward movement of the αC helix. The main energetic bottleneck corresponds to the conformational change in the A-loop, while changes in the DFG motif and αC helix occur before and after A-loop conformational change, respectively. In accordance with this, two intermediate states are identified, the first state just after the DFG flipping and the second state after the A-loop rotation. These intermediates exhibit structural features characteristic of the corresponding inactive and active conformations of other protein kinases. To understand the impact of A-loop phosphorylation on kinase conformation, the free energies of A-loop phosphorylation were determined at several states along the conformational transition path using the free-energy perturbation simulations. The calculated free energies reveal that while the unphosphorylated kinase interconverts between the inactive and active conformations, A-loop phosphorylation restricts access to the inactive conformation, thereby increasing the active conformation population. Overall, this study suggests a consensus mechanism of conformational activation between different protein kinases.

Unravelling patient pathways in the context of antibacterial resistance in East Africa

BackgroundA key factor driving the development and maintenance of antibacterial resistance (ABR) is individuals’ use of antibiotics (ABs) to treat illness. To better understand motivations and context for antibiotic use we use the concept of a patient treatment-seeking pathway: a treatment journey encompassing where patients go when they are unwell, what motivates their choices, and how they obtain antibiotics. This paper investigates patterns and determinants of patient treatment-seeking pathways, and how they intersect with AB use in East Africa, a region where ABR-attributable deaths are exceptionally high.MethodsThe Holistic Approach to Unravelling Antibacterial Resistance (HATUA) Consortium collected quantitative data from 6,827 adult outpatients presenting with urinary tract infection (UTI) symptoms in Kenya, Tanzania, and Uganda between February 2019- September 2020, and conducted qualitative in-depth patient interviews with a subset (n = 116). We described patterns of treatment-seeking visually using Sankey plots and explored explanations and motivations using mixed-methods. Using Bayesian hierarchical regression modelling, we investigated the associations between socio-demographic, economic, healthcare, and attitudinal factors and three factors related to ABR: self-treatment as a first step, having a multi-step treatment pathway, and consuming ABs.ResultsAlthough most patients (86%) sought help from medical facilities in the first instance, many (56%) described multi-step, repetitive treatment-seeking pathways, which further increased the likelihood of consuming ABs. Higher socio-economic status patients were more likely to consume ABs and have multi-step pathways. Reasons for choosing providers (e.g., cost, location, time) were conditioned by wider structural factors such as hybrid healthcare systems and AB availability.ConclusionThere is likely to be a reinforcing cycle between complex, repetitive treatment pathways, AB consumption and ABR. A focus on individual antibiotic use as the key intervention point in this cycle ignores the contextual challenges patients face when treatment seeking, which include inadequate access to diagnostics, perceived inefficient public healthcare and ease of purchasing antibiotics without prescription. Pluralistic healthcare landscapes may promote more complex treatment seeking and therefore inappropriate AB use. We recommend further attention to healthcare system factors, focussing on medical facilities (e.g., accessible diagnostics, patient-doctor interactions, information flows), and community AB access points (e.g., drug sellers).

Assembly of Microbial Communities Engaging in Metabolic Division of Labor in a Diffusion-Limited Environment Is Governed by Metabolic Flux.

Metabolic division of labor (MDOL) represents one of the most commonly occurring interactions within natural microbial communities. Specifically, in a variety of MDOL systems engaged in hydrocarbon degradation, a sequential degradation is performed by several members with final products that are necessary for the growth of each member. In these MDOL systems, each strain catalyzes one or more specific reactions of a multistep metabolic pathway, whose end products are then allocated among the participants. While the benefit allocation is independent of metabolic flux in well-mixed environments, it remains unclear how the benefits are allocated when diffusion is limited. Here, we investigated how MDOL communities assemble in a diffusion-limited environment, by combining mathematical modeling with experimental inquiry using a synthetic consortium engaged in MDOL. Our model analysis in a diffusion-limited environment showed that, when the growth of all populations in the community relies on the final product that can only be produced by the last population, a diffusion gradient of the final products may create a bias favoring the member producing the final products, resulting in a higher relative abundance of the final product producer. Moreover, such asymmetric allocation of the final products is enhanced by both the lower diffusion rate and the higher metabolic flux (i.e., the higher yields of the final products) in the MDOL. Our results show that in a diffusively confined environment, metabolic flux constitutes a determining factor in the assembly of the MDOL community. Together, our findings are critical for a better understanding of how resource-sharing microbial communities are established and should assist in designing such communities for improved biomanufacturing and bioremediation.

Correlating physicochemical and biological properties to define critical quality attributes of a rAAV vaccine candidate

Recombinant adeno-associated viruses (rAAVs) are a preferred vector system in clinical gene transfer. A fundamental challenge to formulate and deliver rAAVs as stable and efficacious vaccines is to elucidate interrelationships between the vector's physicochemical properties and biological potency. To this end, we evaluated an rAAV-based coronavirus disease 2019 (COVID-19) vaccine candidate that encodes the Spike antigen (AC3) and is produced by a commercially viable process. First, state-of-the-art analytical techniques were employed to determine key structural attributes of AC3, including primary and higher-order structures, particle size, empty/full capsid ratios, aggregates, and multi-step thermal degradation pathway analysis. Next, several quantitative potency measures for AC3 were implemented, and data were correlated with the physicochemical analyses on thermally stressed and control samples. Results demonstrate links between decreasing AC3 physical stability profiles, invitro transduction efficiency in a cell-based assay, and, importantly, invivo immunogenicity in a mouse model. These findings are discussed in the general context of future development of rAAV-based vaccine candidates as well as specifically for the rAAV vaccine application under study.

Salient Achievements in Synthetic Organic Chemistry Enabled by Mechanochemical Activation

AbstractAlthough known for millennia, it is only recently that mechanochemistry has received serious attention by chemists. Indeed, during the past 15 years an extraordinary number of reports concerning solid-state chemical transformations through grinding and milling techniques have been recorded. This short review discusses the circumstances that led this renaissance, highlighting the present intense interest in so-called green chemistry, the enabling capacity of mechanochemistry to handle insoluble substrates, and the identification of the profound influence that additives can have on mechanochemically activated reactions. The core of this account focuses on salient developments in synthetic organic chemistry, especially in amino acid and peptide­ mechanosynthesis, the successful employment of mechanochemical activation in combination with asymmetric organocatalysis, the promising combination of mechanochemical activation with enzymatic and whole cell biocatalysis, the remarkable achievement of multicomponent selective reactions via complex, multistep reaction pathways, and the mechanosynthesis of representative heterocycles. The final section comments on some pending tasks in the area, such as scaling-up of milling processes to be of practical use in the chemical industry, the requirement of easier and more efficient control of reaction parameters and monitoring devices, and consequently the careful analysis of additional procedures for a proper understanding of mechanochemical phenomena.1 Introduction2 Brief History of Mechanochemistry3 Milling Equipment and Reaction Parameters4 Attributes of Mechanochemistry That Propelled Its Present Renaissance4.1 Enormous Attention Being Presently Paid to Sustainable Chemistry4.2 Reduced Energy Consumption4.3 Additive-Based Mechanochemistry4.4 Handling of Insoluble Reactants4.5 ‘Impossible’ Reactions That Are Successful by Milling4.6 Successful Handling of Air- and Water-Sensitive Reagents by Ball Milling5 Salient Developments in the Mechanochemical Activation of Synthetic Organic Chemistry5.1 Amino Acid and Peptide Mechanosynthesis5.2 Asymmetric Organic Synthesis and Asymmetric Organocatalysis under Ball-Milling Conditions5.3 Mechanoenzymology5.4 Multicomponent Reactions Activated by Mechanochemistry5.5 Mechanosynthesis of Heterocycles and Modification of Heterocycles6 Future Directions6.1 Scaling-Up Mechanochemical Protocols6.2 Temperature-Controlled Mechanochemistry6.3 Understanding Mechanochemical Transformations6.4 Emerging Mechanochemical Techniques7 Conclusions

Selective control of donor-acceptor Stenhouse adduct populations with non-selective stimuli

Selective control of donor-acceptor Stenhouse adduct populations with non-selective stimuli

A Programmable CRISPR/Cas9 Toolkit Improves Lycopene Production in Bacillus subtilis.

Bacillus subtilis has been widely used and generally recognized as a safe host for the production of recombinant proteins, high-value chemicals, and pharmaceuticals. Thus, its metabolic engineering attracts significant attention. Nevertheless, the limited availability of selective markers makes this process difficult and time-consuming, especially in the case of multistep biosynthetic pathways. Here, we employ CRISPR/Cas9 technology to build an easy cloning toolkit that addresses commonly encountered obstacles in the metabolic engineering of B. subtilis, including the chromosomal integration locus, promoter, terminator, and guide RNA (gRNA) target. Six promoters were characterized, and the promoter strengths ranged from 0.9- to 23-fold that of the commonly used strong promoter P43. We characterized seven terminators in B. subtilis, and the termination efficiencies (TEs) of the seven terminators are all more than 90%. Six gRNA targets were designed upstream of the promoter and downstream of the terminator. Using a green fluorescent protein (GFP) reporter, we confirmed integration efficiency with the single-locus integration site is up to 100%. We demonstrated the applicability of this toolkit by optimizing the expression of a challenging but industrially important product, lycopene. By heterologous expression of the essential genes for lycopene synthesis on the B. subtilis genome, a total of 13 key genes involved in the lycopene biosynthetic pathway were manipulated. Moreover, our findings showed that the gene cluster ispG-idi-dxs-ispD could positively affect the production of lycopene, while the cluster dxr-ispE-ispF-ispH had a negative effect on lycopene production. Hence, our multilocus integration strategy can facilitate the pathway assembly for production of complex chemicals and pharmaceuticals in B. subtilis. IMPORTANCE We present a toolkit that allows for rapid cloning procedures and one-step subcloning to move from plasmid-based expression to stable chromosome integration and expression in a production strain in less than a week. The utility of the customized tool was demonstrated by integrating the MEP (2C-methyl-d-erythritol-4-phosphate) pathway, part of the pentose phosphate pathway (PPP), and the hetero-lycopene biosynthesis genes by stable expression in the genome. The tool could be useful to engineer B. subtilis strains through diverse recombination events and ultimately improve its potential and scope of industrial application as biological chassis.

Possible source and migration pathway for early-summer immigrants of the oriental armyworm, Mythimna separata, arriving in northern Japan

The first generation of the oriental armyworm, Mythimna separata (Walker), arrives every year in northern Japan in mainly late May to early June. Analyses of weather maps suggested that this moth's immigration source could be eastern China, but the accuracy of those analyses was very limited due to the lack of a current standard trajectory analysis. The management of migratory insect pests such as M. separata benefits from the identification of the migration source(s) and pathway(s) of the pests. The present study provides a trajectory analysis for M. separata. Backward trajectories from trap sites in northern Japan were calculated with the HYSPLIT System developed by the U.S. National Oceanic and Atmospheric Administration, taking the flight speed of M. separata and the limitation of low ambient temperature at flight height into account. The ending times of the moth's short and long trajectories were set at dusk on the day before and two days before the possible arrival date, respectively. The results suggested two types of possible migration pathway: a multi-step pathway from Northeast China, the Korean Peninsula, and eastern Russia, which are destination areas of the first-generation's migration, and a direct pathway from seasonal main emigration areas in eastern China such as Jiangsu and Shandong provinces. These findings contribute to our understanding of the migration ecology of M. separata and can be used for the development of methods to predict the migration of this insect.

The Education-Immigration Nexus: Situating Canadian Higher Education as Institutions of Immigrant Recruitment.

A policy shift in the mid-2000s established a pathway between international study and immigration to Canada. Designed to facilitate the settlement of young, highly skilled, and Canadian-trained workers, these pathways are premised on the conception of international students as ideal immigrants. Yet, because higher education institutions enjoy wide latitude when it comes to selecting and admitting international students, this education-immigration nexus has gained scholarly attention and sparked an immigration and settlement debate. What are the implications of having an uncapped temporary foreign worker stream administered by higher education institutions? What are the downstream effects for graduates, employers, and communities as the higher education sector accepts greater numbers of international students every year? What does this mean for the composition of Canadian immigration in the longer term? This paper will highlight the importance and relationship between study, labor market access, and immigration to Canada, consider the roles and responsibilities of higher education institutions in multi-step pathways to immigration, and discuss implications and future strategies which acknowledge the reality of this education-immigration nexus.

Breast Cancer with Brain Metastasis: Molecular Insights and Clinical Management.

Breast cancer is the most frequently diagnosed malignancy worldwide and the leading cause of cancer-related death among women. Brain metastases are a primary contributor to mortality, as they often go undetected until late stages due to their dormant nature. Moreover, the clinical management of brain metastases is complicated by the relevant issue of blood-brain barrier penetration. The molecular pathways involved in the formation, progression, and colonization of primary breast tumors and subsequent brain metastases are diverse, posing significant hurdles due to the heterogeneous nature of breast cancer subtypes. Despite advancements in primary breast cancer treatments, the prognosis for patients with brain metastases remains poor. In this review, we aim to highlight the biological mechanisms of breast cancer brain metastases by evaluating multi-step genetic pathways and to discuss currently available and emerging treatment strategies to propose a prospective overview of the management of this complex disease.

Lithium crystallization at solid interfaces

Understanding the electrochemical deposition of metal anodes is critical for high-energy rechargeable batteries, among which solid-state lithium metal batteries have attracted extensive interest. A long-standing open question is how electrochemically deposited lithium-ions at the interfaces with the solid-electrolytes crystalize into lithium metal. Here, using large-scale molecular dynamics simulations, we study and reveal the atomistic pathways and energy barriers of lithium crystallization at the solid interfaces. In contrast to the conventional understanding, lithium crystallization takes multi-step pathways mediated by interfacial lithium atoms with disordered and random-closed-packed configurations as intermediate steps, which give rise to the energy barrier of crystallization. This understanding of multi-step crystallization pathways extends the applicability of Ostwald’s step rule to interfacial atom states, and enables a rational strategy for lower-barrier crystallization by promoting favorable interfacial atom states as intermediate steps through interfacial engineering. Our findings open rationally guided avenues of interfacial engineering for facilitating the crystallization in metal electrodes for solid-state batteries and can be generally applicable for fast crystal growth.

Stability of Human Telomeric G-Quadruplexes Complexed with Photosensitive Ligands and Irradiated with Visible Light.

Guanine-rich DNA sequences can fold into non-canonical nucleic acid structures called G-quadruplexes (G4s). These nanostructures have strong implications in many fields, from medical science to bottom-up nanotechnologies. As a result, ligands interacting with G4s have attracted great attention as candidates in medical therapies, molecular probe applications, and biosensing. In recent years, the use of G4-ligand complexes as photopharmacological targets has shown significant promise for developing novel therapeutic strategies and nanodevices. Here, we studied the possibility of manipulating the secondary structure of a human telomeric G4 sequence through the interaction with two photosensitive ligands, DTE and TMPyP4, whose response to visible light is different. The effect of these two ligands on G4 thermal unfolding was also considered, revealing the occurrence of peculiar multi-step melting pathways and the different attitudes of the two molecules on the quadruplex stabilization.

Prostate cancer screening-stepping forward with MRI.

To comprehensively review the literature on the integration of MRI as a diagnostic tool in prostate cancer screening and offer practical recommendations for optimising its use. Existing research studies, clinical guidelines and expert opinions were reviewed to support the optimisation standards for MRI use in screening. Consolidated screening principles were used to make appropriate recommendations regarding the integration of MRI into the diagnostic pathway. To strike a balance between the potential benefits of early detection on mortality and minimising the harm of over-diagnosing indolent cancers, it is necessary to have a clear understanding of the context of MRI use. The key to optimisation is patient selections and MRI-targeted biopsies. For men at higher-than-average risk, it is essential to use screening-specific MRI protocols and establish accuracy levels and interpretation criteria. Optimisation of readings by the automation of data acquisition, image quality monitoring, post-processing, radiologist certification and deep-learning computer-aided software is needed. The optimal utilisation of MRI involves its integration into a multistep diagnostic pathway, supported by a quality-assured and cost-effective infrastructure that ensures community-wide access to imaging. MRI in the prostate cancer screening pathway can bring substantial diagnostic benefits. By carefully considering its advantages, limitations and safety concerns and integrating it into a multistep diagnostic pathway, clinicians can improve outcomes while minimising harm to screening participants. The manuscript discusses the role of MRI in prostate cancer screening, highlighting its potential to improve accuracy and reduce overdiagnosis. It emphasises the importance of optimising protocols and integrating MRI into a multistep diagnostic pathway for successfully delivering screening benefits. • Population screening for prostate cancer is a new indication for prostate MRI that allows the detection of high-risk cancers while reducing the need for biopsies and associated harm. • To optimise prostate cancer screening using MRI, it is essential to redefine MRI protocols; establish accuracy levels, reliability and interpretation criteria; and optimise reading (including post-processing, image quality, radiologist certification, and deep-learning computer-aided software). • The optimal utilisation of MRI for prostate cancer screening would involve its integration into a multistep diagnostic pathway, supported by a quality-assured and cost-effective infrastructure that ensures community-wide access to imaging.

Autophagy modulation in breast cancer utilizing nanomaterials and nanoparticles.

Autophagy regenerates cellular nutrients, recycles metabolites, and maintains hemostasis through multistep signaling pathways, in conjunction with lysosomal degradation mechanisms. In tumor cells, autophagy has been shown to play a dual role as both tumor suppressor and tumor promoter, leading to the discovery of new therapeutic strategies for cancer. Therefore, regulation of autophagy is essential during cancer progression. In this regard, the use of nanoparticles (NPs) is a promising technique in the clinic to modulate autophagy pathways. Here, we summarized the importance of breast cancer worldwide, and we discussed its classification, current treatment strategies, and the strengths and weaknesses of available treatments. We have also described the application of NPs and nanocarriers (NCs) in breast cancer treatment and their capability to modulate autophagy. Then the advantages and disadvantaged of NPs in cancer therapy along with future applications will be disscussed. The purpose of this review is to provide up-to-date information on NPs used in breast cancer treatment and their impacts on autophagy pathways for researchers.

Novel Knowledge about Molecular Mechanisms of Heparin-Induced Thrombocytopenia Type II and Treatment Targets.

Heparin-induced thrombocytopenia type II (HIT II), as stated in the literature, occurs in about 3% of all patients and in 0.1-5% of surgical patients. Thrombosis develops in 20-64% of patients with HIT. The mortality rate in HIT II has not decreased using non-heparin treatment with anticoagulants such as argatroban and lepirudin. An improved understanding of the pathophysiology of HIT may help identify targeted therapies to prevent thrombosis without subjecting patients to the risk of intense anticoagulation. The review will summarize the current knowledge about the pathogenesis of HIT II, potential new therapeutic targets related to it, and new treatments being developed. HIT II pathogenesis involves multi-step immune-mediated pathways dependent on the ratio of PF4/heparin and platelet, monocyte, neutrophil, and endothelium activation. For years, only platelets were known to take part in HIT II development. A few years ago, specific receptors and signal-induced pathways in monocytes, neutrophils and endothelium were revealed. It had been shown that the cells that had become active realised different newly formed compounds (platelet-released TF, TNFα, NAP2, CXCL-7, ENA-78, platelet-derived microparticles; monocytes-TF-MPs; neutrophils-NETs), leading to additional cell activation and consequently thrombin generation, resulting in thrombosis. Knowledge about FcγIIa receptors on platelets, monocytes, neutrophils and FcγIIIa on endothelium, chemokine (CXCR-2), and PSGL-1 receptors on neutrophils could allow for the development of a new non-anticoagulant treatment for HIT II. IgG degradation, Syk kinase and NETosis inhibition are in the field of developing new treatment possibilities too. Accordingly, IdeS and DNases-related pathways should be investigated for better understanding of HIT pathogenesis and the possibilities of being the HIT II treatment targets.