Applications range from tunable band gaps, high absorption coefficients, and versatile electronic properties, making them ideal for solar cells to light-emitting diodes and even photodetectors. In this work, we present, for the first time, a nonlinear phenomenological bond model analysis of second harmonic generation (SHG) in tetragonal ABO3 with arbitrary input light polarization. We study the material symmetry and explore the strength of the nonlinear generalized third-rank tensorial elements, which can be exploited to produce a high SHG response if the incoming light polarization is correctly selected. We found that the calculated SHG intensity profile aligns well with existing experimental data. Additionally, as the incoming light polarization varies, we observed a smooth shift in the SHG intensity peak along with changes in the number of peaks. These observations confirm the results from existing rotational anisotropy SHG experiments. In addition, we show how spatial dispersion can contribute to the total SHG intensity. Our work highlights the possibility of studying relatively complex structures, such as ABO3, with minimal fitting parameters due to the power of the effective bond vector structure, enabling the introduction of an effective SHG hyperpolarizability rather than a full evaluation of the irreducible SHG tensor by group theoretical analysis. Such a simplification may well lead to a better understanding of the nonlinear properties in these classes of material and, in turn, can improve our understanding of the photovoltaic performance in ABO3 structures.ABO These octahedrons are interconnected, creating a 3D network whose arrangement depends on the cation sizes and charges. Recently, ABO3 structures in the form of halide perovskites have acted well in outstanding solar cells, which achieved a best efficiency of 25.7% [Perovskite ABOications . These mications . One of ications . The tet3 perovskite structures, which are known to reduce the expected charge-carrier mobility [3 perovskite structure due to the availability of rotational anisotropy second harmonic generation (RASHG) experimental data in Ref. [3 metal halide perovskites, such as methylammonium lead iodide (MAPbI3) and formamidinium lead iodide (FAPbI3), can exist in three different phases depending on the temperature. Above 330 K and 300 K, MAPbI3 and FAPbI3 form a cubic structure, respectively. The tetragonal MAPbI3 phase —which is studied here—exists for a temperature range between 160 and 330 K, whereas for the FAPbI3 perovskite, the tetragonal phase exists between 130 and 300 K. Below 160 K and 130 K, MAPbI3 and FAPbI3 tetragonal phases change into orthorhombic phases, respectively [3, is both intriguing and a subject of ongoing debate because of uncertainties regarding the existence of symmetry breaking inside the material bulk [Another purpose of this work is to apply the nonlinear bond model to investigate the existence of the Rashba effect in the bulk of ABOmobility  and, the in Ref. , which cectively . The occectively . It has ectively . Howeverial bulk .3 perovskite and compare the results with the existing RASHG experimental data. The theoretical results can enhance the PV performance in several ways: (1) Offer a simple way to investigate the molecular structure of ABO3 at the surface/interface, including surface reconstruction/modification [This work can shed light on resolving this debate by performing a theoretical investigation of the nonlinear SHG dipole radiation at the nanoscale within the bulk anfd interface of tetragonal ABOfication  and donofication  for the fication  concernification , which i3 SHG tensor and how to produce the SHG intensity in such structures. The electron charges when driven by an incoming external field will produce linear oscillations. If the intensity of the field is large, as in the case of a femtosecond laser, the charges will also oscillate an-harmonically. Based on the electronic density orbital distribution that can be calculated using density functional theory, the charges are radiating dipoles mainly along the covalent bonds. The electrons also experience damping from their neighboring electrons; hence, the classical dynamical equations should take the form of a dimensional an-harmonically damped-driven oscillation model:Here, we will apply the novel nonlinear theoretical method termed “simplified-bond hyperpolarizability model (SBHM)”. SBHM is a classical phenomenological theory based on a classical Lorentz nonlinear oscillator model proposed in 2002 by Powell and Coworkers . Inspirep and s-polarized incoming fields but instead generalized the model for arbitrary input polarization:z-plane, and s-polarized incoming field, and when p-polarized incoming field.The applied coordinate system in our model is presented in x can be written as Solving Equation  for Δx1 Defining the linear V is the volume, N is the number of atomic units.Therefore, the total polarization can be written as:N and V denote the total SHG dipoles, and V is the volume of the considered computational unit cell. In addition, to model the RASHG experiments later on, the sample is rotated along the z-axis to produce a polar nonlinear intensity plot. In our calculation, the azimuth angle The total macroscopic linear and SHG polarization can be calculated by summing over all dipoles reminiscent of the Ewald–Oseen approach . The forEquation , the SHGIncorporating Equation  into EquThe SHG far field can be obtained via :12)Eff=Eff=(12)E3 structure and study its symmetry properties. Understanding the symmetry of a particular PV structure is very useful in interpreting the results or in simplifying the calculations. We thus begin by analyzing the surface bonds, which only consist of one atomic layer, namely, the layer on the foremost top of the ABO3 3D structure in 3 anions. Due to the nature of the shared electrons in the covalent bonds, the bond unit vector arrows from the A cation and the O3 anions will point toward each other but are not of the same strength due to the different SHG hyperpolarizability In this section, we will present the results and discuss them accordingly. The first step in the SBHM is to develop the bond unit vector for the ABOThe utilization of an effective hyperpolarizability in SBHM has been validated for the case of zincblende structure . In addi3 structure. These consist of the 32 bond vectors aligned slightly horizontally about the z-axis, as given by v2 point group in group theory but now with only one fitting parameter , confirming the inversion symmetry of the ABO3 tetragonal structure. Performing a RASHG for arbitrary polarization can thus present us with more information on the true SHG sources and this work stimulates further experimental work in this area. In addition, our findings will pave the way for a better understanding and modelling of the dynamics of SHG in ABO3 structures. This will be especially valuable in establishing a straightforward connection between the inherent symmetry of the material and the polarization of the driving electric field. It will also enable us to investigate more complex structures, such as surface reconstruction, material impurities, and surface kinetics, using NLO techniques in the future.Having analyzed the third rank SHG tensor, we can obtain the SHG intensity profile for arbitrary input polarization. We choose for the calculation an incoming fundamental-outgoing SHG light angle of ructures . For conEquation  for arbi3 can be studied in a more direct and simplified way via the SBHM. Since SHG is surface sensitive and can detect nanostructural changes, this model can be used to study possible nanoscale impurities and surface reconstruction, which can alter the PV ability. Very recently, SHG has been applied to study whether a spin–orbit coupling mechanism, known as the Rashba effect, inside the bulk of MAPbI3 metal halide perovskite (MHP) exists [3, which we neglected in our model, as can be seen in It is clear from the obtained results that the nonlinear symmetry properties and intensity profile of ABO) exists . The Ras3 and see whether their SHG intensity profile is similar to the surface dipole contribution. The spatial dispersion is straightforwardly calculated by taking the refracted angle close to the normal angle . When we set the angle close to zero, we obtain the 2-fold SHG peak lobe, as given in the bottom left picture in 3 thus invalidating the results in Ref. [However, we have to state that the SHG contribution from the bulk quadrupole and spatial dispersion has not been included in our current discussion. It has been found that these two additional SHG sources can contribute to the total SHG intensity in silicon . Spatial in Ref. . However3 structures. We demonstrate that the SHG dipole contribution originates from the surface and is defined by a set of 20 effective bond vectors. The “generalized” third-rank SHG tensor introduced encompasses the azimuth rotation degree of freedom, revealing that the only requirement for fitting is the effective SHG hyperpolarizability z-axis, is included. Crucial future work entails the inclusion of bulk quadrupole and MA symmetry breaking, vital for determining the potential existence of a static bulk Rashba effect in ABO3 material bulk. Through our work, an enhanced comprehension regarding the dynamics of the SHG peak and phase shift provides deeper insights into SHG in ABO3 structures, with the prospects of investigating surface reconstruction, material impurities, and surface kinetics, all pivotal for optimizing PV efficiency.We have developed previous SHG SBHM models to include arbitrary input polarization for the first time to tetragonal ABO


Owing to the 40-year worldwide efforts for improving diagnosis and therapy for acute myeloid leukemia (AML), the second most common type of leukemia in children, overall survival rates of children with AML have now reached 70% to 80% in developed countries. This review article comprehensively describes the history and advances in the current state-of-the-art risk-stratified therapy for AML in children. However, it is likely that the traditional approaches have already reached their limits, and therefore, novel approaches are absolutely essential. The current state and future directions for incorporating novel molecular-targeted drugs into contemporary therapy through international collaboration are also extensively discussed. These aspects present key solutions for further improvements in outcomes of children with AML.Acute Myeloid Leukemia (AML) is the second most common type of leukemia in children. Recent advances in high-resolution genomic profiling techniques have uncovered the mutational landscape of pediatric AML as distinct from adult AML. Overall survival rates of children with AML have dramatically improved in the past 40 years, currently reaching 70% to 80% in developed countries. This was accomplished by the intensification of conventional chemotherapy, improvement in risk stratification using leukemia-specific cytogenetics/molecular genetics and measurable residual disease, appropriate use of allogeneic hematopoietic stem cell transplantation, and improvement in supportive care. However, the principle therapeutic approach for pediatric AML has not changed substantially for decades and improvement in event-free survival is rather modest. Further refinements in risk stratification and the introduction of emerging novel therapies to contemporary therapy, through international collaboration, would be key solutions for further improvements in outcomes. Acute Myeloid Leukemia (AML) is a form of hematopoietic malignancy characterized by clonal proliferation of immature myeloid cells. As can be seen from the fact that the median diagnostic age of AML is over 60 years old, AML is the most common type of leukemia in adults, whereas it is the second most common leukemia subtype in children, accounting for 20–25% of pediatric leukemia cases, with an incidence of approximately seven cases per 1,000,000 children per year ,4. DespiOverall Survival (OS) rates of children with AML have dramatically improved in the past 40 years, currently reaching 70–80% in developed countries ,15,16,17RUNX1::RUNX1T1)-positive AML in Asian populations (approximately 30% compared to 12–14% of the U.S. or European patients)  concurrent with three days of anthracyclines), but a third drug  is often combined although its role is not fully established . The Japosed AML . Consequ2/dose on days 1 and 21) at the very end of consolidation chemotherapies failed to improve both EFS and OS in the NOPHO-AML2004 study ) ) . Frequenn [TKD]) . Both FL therapy . Sorafen therapy ,84,85. Etwo arms . Pediatr did not . Table 42 venetoclax in combination with HDAC with or without idarubicin was determined to be the recommended phase 2 dose [B-cell/CLL lymphoma-2 (BCL-2) family proteins regulate the intrinsic apoptosis pathway by integrating diverse pro-survival or pro-apoptotic intracellular signals. In AML, increased expression of BCL2 family proteins in leukemic blasts has been reported, and the majority of AML stem cells express aberrantly high levels of BCL2 and are dependent on BCL2 for survival. Furthermore, high expression of BCL2 has been associated with an inferior response to chemotherapy and poor survival among patients with AML. Venetoclax, a selective small-molecule BCL2 inhibitor, has been shown in preclinical studies to induce apoptosis in malignant cells that are dependent on BCL2 for survival. However, the single-agent venetoclax has had only modest activity in AML. Through downregulation of myeloid-cell leukemia 1 (MCL1) and induced expression of the pro-death proteins NOXA and PUMA, azacitidine or cytarabine synergistically inhibits the pro-survival proteins MCL1 and BCL-XL, thereby increasing the dependence of leukemia cells on BCL2. In fact, venetoclax combined with azacytidine or in combination with LDAC significantly prolonged the survival of adult patients with AML unfit for standard chemotherapy ,88. In te 2 dose . The oveKMT2A-r AML accounts for 20–25% of pediatric AML. Recently, the updated retrospective study by the I-BFM was published; notably, nearly 50% of the patients failed the therapy even if their MRD was negative after the second induction [KMT2A-r. Menin is a product of the MEN1 tumor suppressor gene, which binds to the rearranged KMT2A complex and leads to the upregulation of leukemogenic genes (such as HOX and MEIS1), and thus to the subsequent development of acute leukemia. Menin inhibitors have shown selective, profound single-agent activity in KMT2A-r PDX models [NUP98-rearranged AML. NPM1c AML accounts for 20–30% of adult AML and 6% of pediatric AML and is generally associated with good prognosis. NPM1-coding protein nucleophosmin shuttles between the nucleus and cytoplasm during the cell cycle and is involved in diverse cellular processes, such as ribosome biogenesis, centrosome duplication, protein chaperoning, histone assembly, cell proliferation, and regulation of tumor suppressors TP53. However, mutated NPM1 (NPM1c) persists in the cytoplasm, and although the mechanism is not clear (but presumed to be a loss of function), NPM1c is associated with upregulation of HOX genes in a menin-dependent manner [NUP98-rearranged AML accounts for less than 1% of adult AML and 7% of pediatric AML and is associated with unfavorable outcomes. It is known that NUP98 fusion proteins interact with KMT2A chromatin complexes and promote leukemogenesis. Inhibition of menin-KMT2A impairs leukemogenic gene expression and disrupts chromatin binding of menin, KMT2A, and NUP98 fusion proteins in mouse models [KMT2A-r or NPM1c [nduction . Menin iX models . Menin it manner . NUP98-re models . Given tor NPM1c . BecauseCBFA2T3::GLIS2 fusion-positive AML), checkpoint inhibitors, cell-signaling inhibitors targeting MEK (NRAS and KRAS mutations are among the most common mutations in pediatric AML), and epigenetic modifiers  [In addition to the cell signaling inhibitors against FLT3, BCL2, and menin, examples of novel therapies of interest for pediatric AML are immunotherapies  targeting CD123 , CD33, FLT3, or FOLR1 (targeting ibitors) . FinallyA quarter century of global efforts on clinical trials have contributed to improved outcomes for children with AML but are still tentative. Refinement in risk stratification based on leukemia biology and MRD, as well as the introduction of emerging novel therapies to contemporary therapy, through international collaboration, would be a key solution for further improvement in outcomes.


Nuclear Ca2+ is involved in the regulation of cellular Ca2+ homeostasis. However, it remains unclear whether nuclear Ca2+ levels change under skeletal muscle disuse conditions, and whether changes in nuclear Ca2+ levels are associated with nuclear apoptosis. In this study, changes in Ca2+ levels, Ca2+ transporters, and regulatory factors in the nucleus of hindlimb unloaded rat soleus muscle were examined to investigate the effects of disuse on nuclear Ca2+ homeostasis and apoptosis. Results showed that, after hindlimb unloading, the nuclear envelope Ca2+ levels ([Ca2+]NE) and nucleocytoplasmic Ca2+ levels ([Ca2+]NC) increased by 78% (p < 0.01) and 106% (p < 0.01), respectively. The levels of Ca2+-ATPase type 2 (Ca2+-ATPase2), Ryanodine receptor 1 (RyR1), Inositol 1,4,5-tetrakisphosphate receptor 1 (IP3R1), Cyclic ADP ribose hydrolase (CD38) and Inositol 1,4,5-tetrakisphosphate (IP3) increased by 470% (p < 0.001), 94% (p < 0.05), 170% (p < 0.001), 640% (p < 0.001) and 12% (p < 0.05), respectively, and the levels of Na+/Ca2+ exchanger 3 (NCX3), Ca2+/calmodulin dependent protein kinase II (CaMK II) and Protein kinase A (PKA) decreased by 54% (p < 0.001), 33% (p < 0.05) and 5% (p > 0.05), respectively. In addition, DNase X is mainly localized in the myonucleus and its activity is elevated after hindlimb unloading. Overall, our results suggest that enhanced Ca2+ uptake from cytoplasm is involved in the increase in [Ca2+]NE after hindlimb unloading. Moreover, the increase in [Ca2+]NC is attributed to increased Ca2+ release into nucleocytoplasm and weakened Ca2+ uptake from nucleocytoplasm. DNase X is activated due to elevated [Ca2+]NC, leading to DNA fragmentation in myonucleus, ultimately initiating myonuclear apoptosis. Nucleocytoplasmic Ca2+ overload may contribute to the increased incidence of myonuclear apoptosis in disused skeletal muscle.Disuse atrophy of skeletal muscle is associated with a severe imbalance in cellular Ca Disuse conditions, such as spaceflight, prolonged bed rest, and limb breakage, can induce skeletal muscle atrophy, which is characterized by muscle mass loss, cross-sectional area (CSA) reduction, skeletal muscle proteins degradation, and muscle fiber type transformation, resulting in motor function impairing and metabolic disease risk significant increasing ,2.2+ homeostasis occurs during disuse atrophy of skeletal muscle manifested as cytoplasmic Ca2+ overload NE and [Ca2+]NC) in hindlimb-unloaded rat skeletal muscle and examined the capacity and regulation of Ca2+ transporters on nuclear membranes to identify possible reasons for changes in nuclear Ca2+ levels in atrophied muscle. We also explored the potential relationship between nucleocytoplasmic Ca2+ and nuclear apoptosis in atrophied muscle. Therefore, we investigated dysregulation of nuclear Ca2+ homeostasis in disused skeletal muscle to clarify the potential mechanisms and effects of intracellular Ca2+ disturbance in disused skeletal muscle, thereby providing novel targets for the prevention and treatment of disuse atrophy in skeletal muscle.In the present study, we explored CaAll procedures were approved by the Laboratory animal care committee of the P. R. China Ministry of Health. Sprague-Dawley adult healthy female rats, weighing 220−240 g, were provided by Dashuo Company . The individuals were randomly divided into two groups: cage control (CON) group and hindlimb unloading (HLU) group. The CON individual could move freely in a normal cage. The HLU individual was tail suspended for 14 days. The suspending method was referred to Morey-Holton  and modi2+] measurement, WB, ELISA and DNA fragmentation assay, or fixed in 4% paraformaldehyde for TUNEL assay and immunofluorescence staining. After surgery, the individuals were euthanized by injecting an overdose of urethane.At the sampling time, the individuals were anesthetized via intraperitoneal administration of urethane (1000 mg/kg). The soleus (SOL) muscle was carefully separated and quickly surgically removed. The isolated muscle sample was immediately used for NE measurement, incubating with 20 μM Cal-520 AM  containing 0.04% Pluronic F-127  for 20 min, 37 °C. Other part was used for [Ca2+]NC measurement, incubating with 30 μg/mL Cal-520 Dextran  for 20 min, 25 °C. The nucleus and [Ca2+] fluorescence were imaged by laser confocal microscope . The [Ca2+] fluorescence intensity was measured by Fiji-ImageJ software . Mean fluorescence (F) and background value (F0) were measured at a given region of interest (ROI). The content of [Ca2+] was represented by the relative fluorescence intensity (F-F0).The resuspended nuclei were divided into two parts. One part was used for NE levels may affect [Ca2+]NC levels. As such, we used two Ca2+ fluorescent dyes to detect the [Ca2+]NE and [Ca2+]NC levels, respectively. Ca2+ fluorescent dye Cal-520 AM is a methyl acetyl derivative that can penetrate the membrane to combine with free Ca2+ in the nuclear envelope, while Ca2+ fluorescent dye Cal-520 Dextran can pass directly through the nuclear pores into the nucleocytoplasm, where it accumulates and remains for a long time to indicate [Ca2+]NC levels [2+ fluorescent dyes (-AM and -Dextran) to detect [Ca2+]NE and [Ca2+]NC in the nuclei of liver cells [2+]NE and [Ca2+]NC were significantly higher after disuse. Similar to previous studies and our findings that skeletal muscle disuse can lead to severe cytoplasmic Ca2+ overload NE in skeletal muscles after hindlimb unloading.Overall, the marked increase in Ca2+]NC by releasing Ca2+ from the nuclear envelope into the nucleocytoplasm and recycling it back. Two Ca2+ releasing channels are located on INM, the ryanodine receptor (RyR) and inositol 1,4,5-tetrakisphosphate receptor (IP3R), which releases free Ca2+ from nuclear envelope into nucleocytoplasm. There are at least three subtypes of RyR: RyR1 , RyR2 (in cardiac muscle), and RyR3 (mainly in brain) NC overload after muscle disuse.The INM regulates [Can brain) . RyR chan brain) . Kraner n brain) . In the with IP3 ,39. IP3Roverload . In our 2+/calmodulin dependent protein kinase II (CaMK II) and protein kinase A (PKA) in SOL muscle nucleus. CaMK II and PKA can phosphorylate the RyR1 and IP3R1 channels, respectively. This phosphorylation does not open the channels directly, but rather increases the opening probability of the activated channels [2+ release from nuclear envelope. Inhibit the opening probability of channel while increasing the content of channel protein in the skeletal muscle nucleus after hindlimb unloading. We speculate that such opposite changes may be a protective mechanism against excessive increases in nucleocytoplasmic Ca2+.Meanwhile, we have measured the expression of Cachannels ,42. To o2+ into nucleocytoplasm, the INM recycles Ca2+ from nucleocytoplasm back to nuclear envelope, performed via Na+/Ca2+ exchanger (NCX) [2+ concentration, while reactivation of NCX can recovery nuclear Ca2+ concentration [2+ homeostasis. The mammalian NCX family consists of three subtypes: NCX1 is found in neurons, glial cells, cardiomyocytes, and kidney tissues; NCX2 is mainly in brain and NCX3 is mainly in skeletal muscle [2+ uptake from nucleocytoplasm to nuclear envelope. This may also be one of the reasons for increasing nucleocytoplasmic Ca2+ overload.In addition to releasing Caer (NCX) ,43. Prevntration . Nuclearl muscle . We foun2+ uptake from cytoplasm into nuclear envelope on ONM, the enhanced Ca2+ release from nuclear envelope into nucleocytoplasm, and the attenuated Ca2+ recycle from nucleocytoplasm on INM are involved in the Ca2+ overload in the skeletal muscle nuclear envelope and nucleocytoplasm after hindlimb unloading.In summary, the enhanced Ca2+ regulates gene transcription and other processes, and the imbalances in nuclear Ca2+ homeostasis, would lead to some negative, even fatal, effects [2+ overload has been proved [2+ [2+ [2+, leading to an enhancement in DNase X activity, which in turn triggering DNA damage.Nuclear Ca effects . The DNAn proved . DNA bren proved . DNase Xoved [2+ . In our  [2+ [2+ . TherefoCurrently, debate exists regarding whether myonuclei are lost during muscle atrophy. While some studies reported that myonuclei were eliminated ,51, othe2+ level in both nuclear envelope and nucleocytoplasm significantly increases in atrophic SOL muscle after unloading. The expression of nuclear Ca2+-ATPase2 increases to enhance the Ca2+ uptake capacity by outer nuclear membrane, contributing to the increase in nuclear envelope Ca2+ level. The increased expression of RyR1 and IP3R1, as well as the increased levels of CD38 and IP3, may enhance the ability of inner nuclear membrane to release Ca2+ into nucleocytoplasm. Meanwhile, the decreased expression of nuclear NCX3 reduces the ability of inner nuclear membrane to restore nucleocytoplasmic Ca2+. These changes thereby result in the rising of nucleocytoplasmic Ca2+ level. The nucleocytoplasmic Ca2+ overload activates DNase X to degrade DNA, leading to myonuclear apoptosis in hindlimb unloaded skeletal muscle. This study explores the intrinsic causes and results of nuclear Ca2+ imbalance in disused skeletal muscle and explains the mechanisms in the occurrence of nuclear apoptosis. It lays a theoretical foundation for the screening of drug targets based on nuclear calcium regulation and for the development of prevention and treatment measures for disuse muscle atrophy.In the present study, we focused on the changes in nuclear calcium level in skeletal muscle after hindlimb unloading, its mechanisms and possible effects. Our findings are presented in 2+ leakage maybe happened during nucleus isolation attributed to the ionic permeability of nuclear membrane, which would affect the accuracy of the measurement on the content of nuclear Ca2+. Yet, the aim of the present study was to detect the relative difference of nuclear Ca2+ levels in experimental and control groups. The same isolation processes may result in roughly the same extent of the nuclear Ca2+ leakage in the two groups. Moreover, we have carried out a confirmatory experiment, which detecting the phosphorylation levels of CaMK II to confirm the changes of nuclear Ca2+ levels [2+ may have an insubstantial effect, and our results can still indicate the changes of nuclear Ca2+ levels in this study. Nonetheless, the absence of avoiding nuclear Ca2+ leakage still is a limitation of this study. Secondly, we did not assess whether the nuclear Ca2+ levels were in the linear indication range of the Ca2+ indicator Cal-520 AM in this study. Although the results had shown the obvious differences of calcium fluorescence intensity within individuals and groups, this indicator may not be the optimal choice. The appropriate Ca2+ indicator and the necessary measures to prevent nuclear Ca2+ leakage should be found out in future study.Firstly, the nuclear Ca+ levels . The res


Acacia nilotica fruit powder, either alone or in combination, improved growth performance, liver and kidney functions, and zinc retention in tissues.With advances in scientific research and a better understanding of rabbit physiology, emphasis has shifted to optimizing feed nutritional composition and investigating novel nutritional supplements. These supplements are intended to improve performance, immunological function, and to mitigate the negative impacts of heat stress on rabbit production. The current study’s findings reveal that rabbits fed diets containing nanoparticles of zinc oxide and Acacia nilotica fruit powder (ANFP) on production sustainability under hot climatic conditions. Eighty Californian male rabbits aged 40 days old (average body weight 738.5 ± 11 g) were divided into four treatment groups and administered one of the following diets: control diet, Nano-ZnO (50 mg/kg), ANFP (5 g/kg), or a combination of Nano-ZnO (50 mg/kg) and ANFP (5 g/kg) for a period of 60 days. Each of the 20 rabbits used in a treatment was regarded as a replicate. The results showed that adding Nano-ZnO and ANFP individually or in combination to rabbits’ diets improved (p < 0.05) growth performance in comparison to control. In addition, zinc contents in serum or the testis tissues in the Nano-ZnO- and ANFP-treated rabbits were significantly greater (p < 0.05) than those in the control group. In addition, serum levels of creatinine, alanine aminotransferase, and aspartate aminotransferase were decreased (p < 0.05) by supplementation of Nano-ZnO, ANFP, or their combination. Carcass criteria did not differ among the treatments. Overall, the findings of the present study indicate that rabbits fed diets containing Nano-ZnO and ANFP, as well as their combination, showed improvements in growth performance, kidney and liver functions, as well as zinc retention in tissues under hot climatic conditions. The combination of Nano-ZnO and ANFP exhibited the best performance in the rabbits. More research on the synergistic effects of Nano-ZnO and ANFP in the sustainable production of rabbit meat is required.This study aims to examine the effects of supplementing male rabbit diets with nanoparticles of zinc oxide (Nano-ZnO) and  To meet the increasing need for food, as well as to have a positive impact on lowering the heat stress that affects rabbits, it is urgent to find novel natural feed additives for rabbit production. Production of rabbit meat is expanding globally . Given tAcacia nilotica tree is a member of the family Mimosaceae. Acacia nilotica is widely cultivated in subtropical and tropical countries [Acacia nilotica exhibits antioxidant and antibacterial properties [Acacia nilotica fruit powder ethanolic extract showed antioxidant and antibacterial activities in our unpublished in vitro study. There is a lack of knowledge regarding Acacia fruits’ prospective applications in the nutrition of rabbits. The synergistic effects of Zn and Acacia nilotica in reducing the heat stress on rabbits’ performance is worth investigating. Therefore, the objective of this study was to evaluate and compare the effects of either Nano-ZnO or ANFP, alone or in combination, on the growth performance, trace element retention, blood metabolites, and carcass traits of male Californian rabbits.ountries ,20. It ioperties ,22, whicAcacia nilotica tree’s fruits were plucked from a field on a farm in Qena, Egypt as required. Physically and randomly selected samples were collected, promptly weighed, and then left to dry under the sun for 48 h. Dried materials were stored at room temperature in airtight plastic containers with tight-fitting lids after being pulverized to pass through a 1 mm screen with a centrifugal mill  until use. The Acacia nilotica fruits were extracted by hydrodistillation from two batches of dried fruits in a Clevenger-type apparatus for three hours at the Regional Center for Food and Feed, Agricultural Research Center, Egypt. A mass spectrometer detector  and gas chromatography  were used for the analyses of the Acacia nilotica extracts. The GC-MS system included a TG-5MS column . Using the following temperature program, analyses were performed using helium as the carrier gas at a flow rate of 1.0 mL/min and a split ratio of 1:10: 240 °C was reached after rising from 60 °C for one minute at a pace of 3.0 °C per minute. Both the injector and detector were kept at 240 °C. In the two tests, 0.2 µL of the mixes were injected as diluted samples . Electron ionization (EI) at 70 eV with a spectral range of m/z 40–450 was used to produce mass spectra. Utilizing analytical mass spectra , majority of the compounds were identified. The average of two replicates per extract was used to express the relative quantities of active chemicals in Acacia nilotica fruits , total phenols (TP), and total antioxidant capacity (TAC) of the ANFP were analyzed according to El-Chaghaby et al. . The TF The Acacia fruits extract showed the highest levels of TAC, TP, and TF when ethanol was used as the extraction solvent . The amoIn this investigation, 800 mg/kg Zn of nano-ZnO  was used. The particle hydrodynamic diameter measured using dynamic light scattering was less than 100 nm. For aqueous systems, the pH was 7.5 ± 1.5 and the density was 1.7 ± 0.1 g/mL at 25 °C. Scan and transmission electron microscopes (SEM and TEM) were used to analyze the particle’s structural morphology .ad libitum pelleted meals were provided, and automatic nipple drinkers provided access to fresh water. Rabbits were kept on a research farm at the Animal and Poultry Production Department, Agriculture Faculty, South Valley University, Qena, Egypt. The experimental protocol (SVUAGR-3-2023), ensured that the rabbits were treated throughout the trial in accordance with the guidelines for the care of experimental animals, and was approved by the committee of ethics at South Valley University. This study was carried out in compliance with the ARRIVE guidelines. Eighty Californian male rabbits (average body weight 738.5 ± 11) aged 40 days old were divided into four treatment groups and administered one of the following diets: control diet, control diet with Nano-ZnO (50 mg/kg), ANFP (5 g/kg), or a combination of Nano-ZnO (50 mg/kg) and ANFP (5 g/kg). The Nano-ZnO and ANFP were combined with a mineral premix, homogenized, and combined with feed components before being given to the rabbits as pelleted total mixed meals. Twenty replicate cages were used in each treatment. A manual feeder and an automatic drinker were included in the galvanized wire net cages, which were 44 cm in width, 50 cm in length, and 35 cm in height and were used to raise individual rabbits. During the whole 60-day trial period, rabbits were housed in similar management, sanitary, and environmental conditions. Throughout the experiment, The temperature–humidity index (THI) was determined using the following formula:At 40, 70, and 100 days of age, the initial body weight (BW), final BW, and feed intake of rabbits were all measured. Calculations of the daily BW increase and feed intake between 40 and 70, 70 and 100, and 40 and 100 days of age were performed. The feed conversion ratio (FCR) was estimated by subtracting the average daily body weight growth from the daily feed consumption. Every case of mortality and every instance of diarrhea was recorded as it happened.n = 20), and they were immediately placed in freezers at –20 °C for Zn content analysis. With the use of an atomic absorption spectrophotometer , the Zn concentrations in the rabbits’ control diet, serum, liver, kidney, and testis were determined.Samples of the testicles, liver, and kidneys were removed from dead rabbits  were used to measure serum creatinine, urea, aspartate aminotransferase (AST), and alanine aminotransferase (ALT).After fasting for 12 h, 20 rabbits per treatments were slaughtered (n = 20) on termination of the experimental period. The animals were weighed at slaughter to ascertain their live BW and killed humanely in accordance with halal slaughtering procedures. The carotid artery, jugular vein, trachea, and esophagus were all severed. The skin, genitals, urinary bladder, gastrointestinal tract, and distal section of the legs were taken from the slaughtered rabbits after they were bled. After emptying, the head and entire gastrointestinal system were weighed and expressed as g/kg slaughter weight. Furthermore, the weight of the entire gastrointestinal system was determined. Testes, liver, spleen, heart, lungs, kidneys, and perirenal and scapular fat were all weighed and represented as g/kg of carcass. For the dressing percentage, the carcass, head, liver, kidneys, and heart were all considered. Dressing was expressed as g/kg and proportionate to the live weight at slaughter.p < 0.05 was used to declare significance.Using SAS 9.2′s general linear model (GLM) approach and a completely randomized design, the statistical analysis was carried out. All analyses used the pens as the experimental unit. All data were evaluated for normal distribution (W > 0.05) using the Shapiro–Wilks test. Then, Duncan multiple range tests were employed to compare means after one-way ANOVA was carried out using the SAS 9.2  statistip < 0.01) the body weight of rabbits at 70 and 100 days of age. Likewise, body weight gain was increased by supplementation of Nano-ZnO and ANFP individually or in combination, in comparison to control diet throughout the experiment phases . In addition, feed conversion ratio was improved (p < 0.01) by supplementation of Nano-ZnO and ANFP individually or in combination in rabbits’ diet during 40–70, 70–100, and 40–100 days of age. When Nano-ZnO and ANFP were added to the diet of Californian males during the 70–100 day and 40–100-day periods, respectively, feed intake improved (p < 0.05) but did not differ between treatments during the first period at 40–70 days of age. Overall, the combination of Nano-ZnO and ANFP was found to provide the best growth performance, including body gain and feed conversion ratio  than those in the control group (p < 0.001) Zn contents in the testis than those consuming the control diet (p < 0.05) by supplementation of Nano-ZnO, ANFP, or their combination compared to the non-supplemented control group (p < 0.05) by supplementation of Nano-ZnO, ANFP, or their combination compared to the non-supplemented control group (Serum Zn concentrations in the Nano-ZnO- and ANFP-treated rabbits were greater (ol group . Rabbitsrol diet . Howeverrol diet . In addiol group . Likewisol group .p < 0.05) the dress-to-live body weight ratio compared to the control group under hot climatic conditions. The testes-to-BW ratio increased with the Nano-ZnO and ANFP individually or in combination compared to the control group. There were no observed differences in head, liver, heart, spleen, and kidneys to live BW ratio due to Nano-ZnO and ANFP individually or in combination compared to control.The effects on carcass criteria of rabbits due to Nano-ZnO and ANFP individually or in combination are presented in Acacia nilotica. The antioxidant and antibacterial properties of Acacia nilotica are mostly attributed to its phytochemical composition, particularly polyphenols [Acacia nilotica fruits may be a significant predictor of its antioxidant capacity [The literature is lacking in information regarding the potential usage of Nano-ZnO and ANFP alone or in combination in rabbit feeding and the impact on the sustainability of production of Californian male rabbits under hot climatic conditions. Therefore, comparisons were made with earlier works, as well as with other studies that investigated ZnO or yphenols ,22, whiccapacity . The phyAcacia nilotica bark extract feeding [In this study, adding Nano-ZnO and ANFP individually or in combination to Californian male diets improved the body weight, body weight gain, and feed conversion ratio of rabbits under hot climatic conditions. This improvement was partly related to a tendency to increase feed intake. In addition, Nano-ZnO and ANFP combination showed the best growth performance, which may be mostly attributable to the synergistic effects of Zn and ANFP in reducing the heat stress on rabbits. The primary phytochemical components of ANFP, particularly its polyphenol concentrations, are responsible for its capacity to enhance growth performance, stimulate appetite, and increase feed intake ,18. In t feeding . Additio feeding . SupplemAcacia nilotica exhibited excellent antimicrobial activity against resistant bacteria, were harmless for animal cells, and were long-lastingly stable [The Zn concentration in the blood and testes was higher in the rabbits’ consuming diets supplemented with Nano-ZnO and ANFP separately or in combination, but not different in terms of liver and kidney tissues. Zn has an impact on the motility of spermatozoa and is crucial for the development of the testes, spermatogenesis, and capacitation . Dietaryy stable . This miAcacia nilotica has shown significant antioxidant activity due to its high concentration of polyphenolic components, such as catechins, which are known to have antioxidative and anti-inflammatory effects [Acacia nilotica, according to Kannan et al. [Acacia nilotica has liver and kidneys protective properties.In the present investigation, Nano-ZnO- and ANFP-treated rabbits had decreased ALT, AST, and creatinine levels in their serum compared to control rabbits. The fact that Zn is a necessary component of roughly 200 Zn metalloenzymes and is present in a variety of enzymes that support the structural integrity of proteins may be the cause of these improvements in male rabbits’ blood biochemistry, including their liver and kidney functions ,27. For  effects  and thusn et al. , and theFurthermore, our results showed that diets supplemented with Nano-ZnO and ANFP individually or in combination increased the dressing and testes percentages without any difference in liver, kidney, and heart percentages compared to the control group under hot conditions. The control group’s live body weight (n = 20) was slightly higher before slaughter following a 12 h fasting interval  than theUnder high temperature conditions, adding Acacia bark extract to rabbit rations significantly boosted dressing percentages without any significant effects on internal organs . OverallThe present study indicates that rabbits fed diets containing Nano-ZnO and ANFP, either alone or in combination, showed improvements in growth performance, liver and kidney functions, as well as Zn retention in tissues. The combination of Nano-ZnO and ANFP can be applied as feed additives in rabbit male diets, as it exhibited the overall best performance in the present study. Future systematic studies on the beneficial synergistic effects of Nano-ZnO and ANFP in the production of male and female rabbit meat are needed.


We study the liquid phase behavior of ternary mixtures of monodisperse hard spheres in solution. The interactions are modeled in terms of the second virial coefficient and can be additive hard sphere (HS) or non-additive hard sphere (NAHS) interactions. We give the set of equations that defines the phase diagram for mixtures of three components. We calculate the theoretical liquid–liquid phase separation boundary for two-phase separation  and, if applicable, the three-phase boundary, as well as the plait points and the spinodal. The sizes of the three components are fixed. The first component (A) is the smallest one, the second component (B) is four times the size of the smallest component, and the third (C) component is three times the size of the smallest one. The interaction between the first two components is fixed, and this AB sub-mixture shows phase separation. The interactions of component C with the other two components are varied. Component C can be compatible or incompatible with components A and B. Depending on the compatibility of the components, the phase diagram is altered. The addition of the third component has an influence on the phase boundary, plait points, stability region, fractionation, and volume ratio between the different phases. When all sub-mixtures  show phase separation, a three-phase system becomes possible when the incompatibility among all components is high enough. The position and size of the three-phase region is dependent on the interactions between the different sub-mixtures. We study the fractionation off all components depending on specific parent concentrations. A lot of research, both theoretical and experimental, has been focused on the phase behavior of two components in solution. However, most products consist of more components. The investigation of multicomponent phase behavior has a considerable number of challenges. From an experimental point of view, the number of samples to prepare to obtain an insight into the phase behavior at different concentrations in the phase diagram increases significantly with the number of considered components. From a theoretical point of view, the number of pairwise interactions for a mixture with n components taken into account increases according to nn + 1)/2. The number of interactions increases even more when higher-order interactions are taken into account. Depending on the interactions between all the components, the theoretical number of possible phases that can be formed will increase as well, as follows from the Gibbs phase rule. The Gibbs phase rule gives the relationship between the number of possible phases and the degrees of freedom (dof) for a system: dof = n + 2 − f, in which n is the number of components and f is the maximum number of phases in equilibrium. The degrees of freedom for the system are, for example, the mole fraction of the components, the temperature, or the pressure. When looking at multicomponent systems, one may distinguish chemically different components and chemically equivalent components that only differ in size. The latter are often met in experimental conditions since many components show polydispersity in their size  /2. The n,4. In thDepletion interactions in systems with more than two components become more complicated than for a two-component system. This has been shown by Ji and Walz , who invSome experimental work on mixtures with more than two components that demix into multiple-phase systems has focused on methods to aid in the partitioning of proteins or cell contents. Each of the phases in these systems has a different density and relatively low surface tension, aiding in the separation. An example of such a system is the system containing the high-molecular colloids dextran, ficoll, and poly-ethylene-glycol ,8. SimilA more in-depth experimental study on the phase behavior and phase diagram of a mixture of three components and the three-phase dynamics was performed by Beck-Candanedo et al. . They stThe work on three-component systems paves the way for analysis of systems containing multiple-component systems. This may be important, for example, for cellular systems. Cells contain a large number of components in solution, and it has been hypothesized that demixing and phase separation leads to organization within cells . It shouAnother important application area is the phase behavior of practical industrial systems, such as in cosmetics and food systems, which consist of various types of polymers, each of which is, in fact, polydisperse .For multi-component systems, Sear and Cuesta  investigIn order to identify interesting characteristic behavior of such multicomponent systems, one could also attempt the same second-order virial approach, instead of randomly attributing virial coefficients as in the random matrix theory approach. One can then use experimentally accessible virial coefficients between the components and solve the equations numerically. Our work is aimed at this approach, starting with three-component systems in this article, which is worked out for mixtures with more components in detail elsewhere . It is nThe descriptions of characteristic behavior, such as those by Sear and Cuesta , are difInterestingly, work has been done on ternary additive hard sphere systems , though In our current study, we aim to obtain a better understanding of how the addition of a third component influences the phase behavior of two components that show phase separation. We will study the position of the phase separation boundary, the spinodal, the plait points, and the fractionation. We model the interactions between the components using a virial approach up to second order. Then, we describe the equations for the stability boundary and the spinodal, the equations for the plait point line, and finally the equations defining the phase boundary. With our expressions, we have enough to calculate the full phase diagram for a variety of mixtures. We do this until we reach a summed concentration of 0.5. We add a third component C to a binary mixture AB that phase separates. First, the component C is compatible with both A and B. In the second scenario we study, C is compatible with A or B, and incompatible with the other one. In the third scenario, C is incompatible with both A and B. Subsequently, we investigate the concentration of each component in the phase-separated mixtures at specific parent concentrations. In the last section, we compare three-component systems with a different particle size and different interactions that show three-phase separation.We show the equations used for the calculations of the ternary phase diagram of the different studied systems: the set of equations defining the stability boundary, the plait points, and phase boundaries of a mixture. All sets of equations are solved in MATLAB R2017b. For a more detailed derivation of the equations, we refer to  and refeT, can be written as a virial expansion, similar to the virial expansion of the universal gas law for real gases [β = ; k is Boltzmann’s constant; T the temperature; ρ the number density of the solute component, V; and B2 and B3 the second and third virial coefficients of the solute component. The second virial coefficient accounts for the increase in osmotic pressure due to particle pairwise interaction. The third virial coefficient accounts for the interaction between three particles in a variety of configurations. The equation can be expanded for higher densities with iB, the ith virial coefficient of the solute, which accounts for the interaction between i different solute particles.The osmotic pressure, Π, of a solute in a fluid, at a temperature al gases :βΠ = ρ +sµ is the chemical potential of the solvent, ijW (r) is the interaction potential between the particles i and j, and r is the distance between the center of the particles. For additive hard sphere (HS) interaction, the interaction potential for two particles (of the same species or different species) is, under constant chemical potential of the solvent, simplified to:ijσ = (iσ + jσ)/2, the distance between the centers of the two particles, and iσ and jσ are their respective diameters. The assumption of the chemical potential of the solvent remaining constant holds when the solute particles approach one another under the condition of constant osmotic pressure of the solution and constant partial volume of the solvent.In this work, we will limit the virial expansion to the second virial coefficient, which is given by :(2)B2T,μijσ = ((iσ + jσ)/2)(1 + ∆), in which ∆ (≥−1) accounts for the non-additivity of the interaction between the particles. When ∆ > 0, the distance of closest approach of both spheres increases, and when ∆ < 0, the distance of closest approach decreases compared to that due to HS interaction only. For additive hard sphere interaction, ∆ = 0. We note that for hard spheres and non-additive hard spheres, the virial coefficient does not depend on temperature, nor on the density of the spheres.For non-additive hard spheres (NAHS), the distance of the closest approach of the centers of the two particles of different species can be closer or further than the distance between their centers . The clon distinguishable components in a solution, there are two main types of two-particle interactions that can occur: between particles of the same species and particles of different species. For the system we are studying, n = 3 , and xyB the second virial coefficient of particles of different species, which can be HS or NAHS.For the second virial coefficient given by Equation (2), using the interaction potential defined in Equation (3), we find:The general equation for the osmotic pressure for a dilute mixture of three components, A, B and C, limited to the second virial coefficient, is given by :(6)βII=ρiµ, the chemical potential ) for component i, is given by:The differential of the free energy of a mixture is given by :(7) dA=−For a mixture with three distinguishable components, the second partial derivatives can be represented by a 3 × 3 matrix of the first partial derivatives of the chemical potential of each component.This results in the following stability matrix:The mixture is stable when all eigenvalues are positive . When, oIn a binary mixture, the critical point is a stable point that lies on the stability limit   and wherdet(M1) = 0, which is the equation for the spinodal. The other criterion is based on a characteristic at the critical point. For a multicomponent system, using Legendre transforms, one finds det(M2) = 0 [There are two criteria that can be used to find plait points. The first one is (M2) = 0 ,28, withM2 is matrix M1 with one of the rows replaced by the partial derivatives of the determinant of matrix M1. Note that it does not matter which row of the matrix is replaced.Matrix We find the following:iP can be found using the following equation [Mii)1,Pi=−det(M1) = 0 and det(M2) = 0), especially when all three components are incompatible, has solutions that are not on a phase boundary. We consider this a mathematical solution to the set of equations, but not a physical solution for a plait point, since plait points are defined to be concentrations on the phase boundary where two or more phases become indistinguishable. Therefore, these solutions will be ignored when depicting the phase diagram.We note that for some of the mixtures, the set of equations , results in the following set of equations :(14)  βIWith Equation (14), we have 2 × 3 + 1 unknowns and 2 × 3 + 1 equations for mixtures that separate into two phases. For mixtures that demix into three phases, we will have 3 × 3 + 2 unknowns and 3 × 3 + 2 equations. Therefore, the set of equations given by Equation (14) allows for calculating the concentration of each component in each of the phases for any given parent concentration.A, B, and C) with non-additive interactions. For all particles, the concentrations are expressed as a dimensionless parameter according to iη = πρA, B, or C. If we refer to η, we imply the sum over iη over each i that is relevant to the specific case.We calculate the phase diagram for a variety of ternary mixture of hard spheres . To this mixture, we add a component C, with size ratio ACq = Aσ/Cσ = 1/3. The size of C is therefore in-between the sizes of A and B. We vary the non-additivity parameters (∆AC and ∆BC) between components A and C, and B and C, respectively, from ∆ = −0.1 to ∆ = 0.1, simulating a range of thermodynamic compatibility and incompatibility between the three components. The phase diagram of this binary mixture of A and B can be found in We define a specific binary mixture of components η < 0.5). An easy way to check for the possibility of phase separation for binary mixtures is by using the following parameter [AB has a critB of 6.76. For mixtures with more than two components, we have to find another criterion to check for the possibility of two- or three-phase separation. We will therefore first investigate a number of different mixtures depending on the interactions in the binary sub-mixtures. We start with mixtures for which the component C does not show any phase separation at concentrations η < 0.5 with A or B. In this case, C shows thermodynamic compatibility with both A and B. In the next section, C is incompatible with A or B. The binary mixture AC or BC will phase separate and have a critical point at concentrations η < 0.5. In the third section, C is incompatible with both A and B. The binary mixtures AC and BC both have a critical point at concentrations η < 0.5. For this type of mixture, we hypothesize that it becomes possible that the mixture will phase separate into more than two phases, inspired by the hypothesis put forward by the work of Mace et al. [Mixtures are able to demix when the stability matrix (Equation (9)) has a negative eigenvalue . In this case, the sub-mixture BC will not demix at any concentration, because critBC <B 1 ; however, the critical point is at unattainably high concentrations (crit >η 1). For the ternary mixture, we see that the addition of the third component C to the binary mixture AB has little effect on the phase behavior of the two components. With increasing concentrations of C, the plait points shift to slightly higher concentrations of B, and the spinodal shifts towards lower concentrations of B at higher concentrations of A. There is no difference in the position of the binodal. With increasing concentration of C, the two phases will have the same volume fraction (α) and the same demixing of A and B, independent of C. See also C is present in both phases with concentrations similar to the concentration in the parent phase. For this system, the tie lines show very little rotation, so they are relatively parallel to the xy-plane.In the first mixture , we set  < 1 see ; the sizA and C the same, but we increased the non-additivity parameter between B and C to hard sphere interaction (∆AC = −0.1 and ∆BC = 0). Now, it is theoretically possible for B and C to phase separate (critBC >B 1)  see ; howeverA and B with the addition of the third component C is an indication that the volumes of both phases will be different for specific mixtures of A and B with the addition of C. Not only will the volumes change, but the preferential fractionation of the components in both phases will also be different. The more the plait points shift to different concentrations, the more the tie lines between phases will show rotation with the addition of the third component. This can be seen in C has a preference of going to the top phase (phase enriched in A), and the volume fraction (a) of the bottom phase (phase enriched in B) decreases slightly with an increased addition of C. The tie lines in this system start to show some rotation.The fact that the plait points shift to different concentrations of AC = 0 and ∆BC = −0.1. The sub-mixture AC can phase separate at concentrations η < 0.5, but the sub-mixture BC cannot demix at all . The sub-mixture AC can phase separate at concentrations η < 0.5, but the sub-mixture BC cannot demix at this low concentration, because the size difference between B and C is not large enough  of the bottom phase increases as well , the sections of plait point line meet each other at even lower concentrations  of the bottom phase increases more upon phase separation compared to the previous mixture.With increased incompatibility between trations . In the trations b, the crparation , bottom;A and C have a negative non-additivity parameter, and B and C have a positive non-additivity parameter (∆AC = −0.1 and ∆BC = 0.1). The sub- mixture AC does not have a critical point at physically relevant concentrations to phase separate; however, the sub-mixture BC does phase separate. For the ternary mixture, the plait points shift towards higher concentrations of B with increasing concentration of C, until the critical point of the sub-mixture BC is reached. With decreasing concentration of A, the binodal and spinodal bend towards the yz-plane, until the surfaces meet with the binodal and spinodal of the binary BC mixture. In the cross-section  of the top phase increases  all show phase separation, as can be seen from both the binodal and the spinodal surfaces shifting towards the xz-plane and yz-plane  all show phase separation. For this mixture, both the binodal and the spinodal surfaces shift towards the xz-plane and yz-plane as well. In this case, the incompatibility between all three components is large enough for the mixture to demix into three phases at physically relevant concentrations. The lowest concentration of the three-phase boundary is at critη = . This point is on the spinodal, the plait point line, and also the binodal. This is a special plait point, where all three phases become indistinguishable. Small deviations in one of the concentrations of the three components will result in the formation of one homogeneous phase, two phases , or three phases. There are again two sections of plait point lines for this mixture, just as in some of the mixtures in the previous section. These two sections of plait point lines do not meet each other, not even at very high concentrations of C, since one section terminates at the three-phase boundary. The two-phase boundary has a bend where the surface meets with the three-phase boundary. Depending on the concentrations of A, B, and C, the two phases will have very different fractionation.The three sub-mixtures of the mixture in C and two components A and B that phase separate. In this section, we will qualitatively compare the phase behavior of the different mixtures at specific parent concentrations of the different components to obtain more insight into the phase separation dynamics. In all the fractionation figures, each component has a different color: component A is red, component B is green, and component C is blue . For all mixtures, component A, being the smallest component in the mixture, preferentially goes to the top phase, while component B, the largest component in the system, remains at the bottom. The volume fraction of the bottom phase (α) is largest for the mixtures from A and C were more incompatible than B and C, as can be seen in critB values. Component C preferentially goes to the bottom phase (more blue in bottom phase compared to top phase) at this parent concentration, and the volume fraction of the bottom phase correspondingly increases.In the previous sections, we saw the phase diagrams of nine different mixtures for which we altered the interactions between a third component blue see . DependiC , C prefers the top phase, but for the two mixtures with the highest incompatibility between A and C and low to moderate incompatibility between B and C , A, component C demixes from the top phase, forming the new middle phase. Finally, we look at a parent concentration with a lower amount of B , AB does not show phase separation yet. However, due to the addition of component C, some of the mixtures demix at this parent concentration. This is especially true for the mixtures with more compatibility between B and C and higher incompatibility between A and C, the mixtures from A and C; however, due to the increased incompatibility between B and C, the mixture remains stabilized. When a mixture demixes into two or more phases, the concentration of one or more components is higher in one of the phases compared to the other, while each of the phases has a smaller volume compared to the parent volume. Systems with less incompatibility among some of the components tend to demix at slightly lower concentration compared to systems with high incompatibility. This was also shown experimentally by Johansson and Walter [When increasing the concentration of component d Walter  for the C and the non-additive interaction parameters ∆AC and ∆BC to investigate the dynamics in three- phase systems. First, we changed the size ratio between A and C. For the mixture in ACq = 1/5, and the non-additive interactions between C and the other components are ∆AC = 0.1 and ∆BC = 0.1 ; however, their respective fractions are flipped due to their flip in size ratio. Compared to η. Since component C is now the largest component in the system , this component preferentially goes to the bottom phase, component B preferentially goes to the middle phase, and component A is most abundant in the top phase. When we navigate through the phase diagram on a straight line with increasing component A while keeping the concentration of components B and C the same, the system first demixes into a large top phase enriched in component A and B and a small bottom phase enriched in component C. This is due to the higher incompatibility of C with A compared to the incompatibility between A and B. When more of component A is added, component B demixes from the top phase, forming the middle phase.In the previous sections, we saw one three-phase system. We changed the size of crit see , we see C as in the first series of mixtures (ACq = 1/3), and we increased the incompatibility between A and C. Compared to η. When gradually increasing the concentration of component C, the two-phase system of one phase enriched in A (the top phase), and the other phase enriched in B. When reaching the three-phase boundary, component C demixes from the top phase, forming the middle phase, which is enriched in component C.In Experimental three-phase systems of, for example, the colloids dextran, ficoll, and polyethylene-glycol  and dextC to a binary mixture AB that demixes has a different impact depending on the pairwise compatibility between the third components and the two other components. Depending on the pairwise interactions for the three components, the volume fractions and compositions of the three phases are altered. If the third component is compatible with both components, the phase boundary and spinodal are nearly vertical surfaces. The third component therefore does not decrease the incompatibility between the components A and B. The component C is present in both phases in the same amount, and the volume fraction between the phases is not altered by the addition of the third component. The plait point line of the mixture will shift to lower or higher concentrations of the components A and B with increasing concentration of C depending on the interaction of C. The sum of the volume fractions of A and B is, however, always equal or lower compared to the sum of the volume fractions at the critical point of the binary mixture.The addition of a third component C is incompatible with one of the components A or B, the binodal and spinodal will bend towards one of the planes (either the xz-plane or yz-plane) of the phase diagram. The plait point line forms a curve that connects the critical points of the binary mixtures that demix. When all three components are incompatible, it becomes possible for the mixture to demix into three phases. The lowest concentration of the three-phase boundary also lays on the spinodal, and for some of the mixtures on the two-phase boundary as well. For these mixtures, small perturbations in the concentrations of one of the components at this critical point will result in one homogeneous phase, two phases, or three phases. Depending on the different pairwise interactions between the components, the three-phase boundary has a different shape and rotation.When


ALDH1A1 and CFD were associated with the prognosis of AML. Four FDA-approved drugs were selected, and a top-ranked drug was identified for each biomarker through molecular docking studies. The top-ranked drugs were further confirmed by molecular dynamic simulation that revealed their binding stability and confirmed their stable performance. Therefore, the drug compounds, enasidenib and gilteritinib, can be recommended as the most effective drugs against the ALDH1A1 and CFD proteins, respectively.Acute myeloid leukemia (AML) is a blood cancer caused by the abnormal proliferation and differentiation of hematopoietic stem cells in the bone marrow. The actual genetic markers and molecular mechanisms of AML prognosis are unclear till today. This study used bioinformatics approaches for identifying hub genes and pathways associated with AML development to uncover potential molecular mechanisms. The expression profiles of RNA-Seq datasets, GSE68925 and GSE183817, were retrieved from the Gene Expression Omnibus (GEO) database. These two datasets were analyzed by GREIN to obtain differentially expressed genes (DEGs), which were used for performing the Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, protein-protein interaction (PPI), and survival analysis. The molecular docking and dynamic simulation were performed to identify the most effective drug/s for AML from the drug list approved by the Food and Drug Administration (FDA). By integrating the two datasets, 238 DEGs were identified as likely to be affected by AML progression. GO enrichment analyses exhibited that the upregulated genes were mainly associated with inflammatory response (BP) and extracellular region (CC). The downregulated DEGs were involved in the T-cell receptor signalling pathway (BP), an integral component of the lumenal side of the endoplasmic reticulum membrane (CC) and peptide antigen binding (MF). The pathway enrichment analysis showed that the upregulated DEGs were mainly associated with the T-cell receptor signalling pathway. Among the top 15 hub genes, the expression levels of  Acute myeloid leukemia (AML) is a hematopoietic malignancy (cancer) in which too many immature blood-forming cells are accumulated in the bone marrow that also interferes with the production of normal blood cells, such as red blood cells, platelets, white blood cells, and other components . It is tA detailed understanding of the molecular mechanism for AML pathogenesis is likely to provide a rationale for developing and designing an appropriate therapy. FDA approved a list of drugs for medicating AML, but doctors speculate about prescribing the most effective drug from the list. However, it is a bit difficult for a doctor to select a drug having the most effectiveness. So, the doctors are to prescribe a drug for a patient as if they experiment on the patient. Our study is aimed at exploring the prognostic biomarkers and the molecular pathways related to AML and thereby identifying the best-matched specific therapeutic compounds for the appropriate treatment of AML through molecular docking and molecular dynamic (MD) simulation.The workflow of the present study is presented in https://www.ncbi.nlm.nih.gov/geo/) and selected two datasets, GSE68925 [de novo AML patients, and three  were healthy samples taken as control. The remaining samples were excluded as they are refractory secondary AML  and refractory/relapsed AML . The selected datasets were analyzed from Homo sapiens, and the tissue system and cell type were also relevant to AML for both datasets. Detailed information about the data sets is shown in This study used the keyword “Acute Myeloid Leukemia” and searched on the NCBI's GEO database  for AML samples from two RNA-seq datasets. The distribution of the DEGs in the datasets was presented as a Venn diagram using the Venny online tool [http://www.bioinformatics.com.cn/srplot) [GREIN (GEO RNA-seq Experiments Interactive Navigator)  is an inine tool . Volcano/srplot) .https://david.ncifcrf.gov/) [p value < 0.05 was defined as significant enrichment.The Database for Annotation, Visualization, and Integrated Discovery   is an onhttps://string-db.org) database was used to build the protein-protein interaction (PPI) network by setting up their interaction score > 0.4 [The STRING  was usehttp://gepia2.cancer-pku.cn/) is an interactive bioinformatics online tool used for investigating the RNA-sequencing data from the TCGA and the GTEx projects [p value was < 0.05. The possible effects of hub genes were assessed using the overall survival (OS) study [Gene Expression Profiling Interactive Analysis  study . These hhttp://gepia2.cancer-pku.cn/) includes different functions such as tumor and normal differential expression analysis, profiling of cancer types or pathological stages, and survival analysis of patients. GEPIA was used to exhibit the box plot for revealing the results to validate and analyze the expression of key genes. Log (FC) > 1 and p value < 0.05 were set to screen the data's validity.GEPIA  and CFDThe protein structures were preprocessed by PyMOL software (version 2.5) , where wX = 44.018, Y = −14.84, and Z = 19.702; and dimensions are X = 76.3418565941, Y = 66.6992212296, and Z = 55.7972505379 . For CFD, the center points of the box were X = −1.4345, Y = −0.0011, and Z = 13.0908; and dimensions are X = 43.7299689674, Y = 43.2379607773, and Z = 55.4447107124 . Molecular visualization is important in modelling analysis after docking the selected compounds with the target proteins. The docking results were visualized and analyzed using BIOVIA Discovery studio client 2021 [The grid box was generated, and for ALDH1A1, the center points of the box were ent 2021 .http://scidavis.sourceforge.net. All snapshots were then subjected to YASARA software's MM-Poisson–Boltzmann surface area (MM-PBSA) binding free energy calculation using Formula  or external  changes the structure of the ligand involved in the motion (rotations and translations). It is also more realistic to evaluate the molecular recognition between the ligand and the target protein. Due to the significant energy dissipation for each conformation, this technique takes a longer time to determine the best-docked conformer. Fast optimization methods and grid-based tools have largely transformed this flaw in recent years, making simulation more user-friendly . The mol Formula  below [3Here, BFE: binding free energy; EpotR: EpotReceptor; EsolR: EsolvReceptor; EpotL: EpotLigand; EsolL: EsolvLigand; EpotC: EpotComplex; EsolC: EsolvComplex.With AMBER 14 as the force field, the MM-PBSA binding energy was estimated in this case using built-in YASARA macros, where bigger negative energies indicate better binding .GSE68925 and GSE183817 were investigated to identify DEGs by comparing blood samples among healthy and AML patients. A total of 1965 differential genes in the GSE68925 dataset (1221 upregulated and 744 downregulated genes) and 1449 differential genes in the GSE183817 dataset (614 upregulated and 835 downregulated genes) were identified by using GREIN . The uprThe aim of the integration of DEGs is to find out the common DEGs from two datasets. The two datasets were integrated using the online tool Venny 2.1.0 , which iGO is a technique for locating classes of genes or proteins that are overrepresented in a large collection of genes or proteins and may be related to disease characteristics. DEGs are categorized according to their biological process (BP), cellular component (CC), and molecular function (MF) in GO analysis. The functional processes of the DEGs were performed using GO analysis, shown in Tables In BP, the upregulated DEGs were enriched in the inflammatory response, immune response, CC, extracellular region, extracellular space, plasma membrane, endosome membrane, and MF serine-type endopeptidase activity. In BP, the downregulated DEGs were enriched in the T cell receptor signalling pathway and costimulation, regulation of gene expression, embryonic hematopoiesis, antigen processing and presentation of peptide or polysaccharide antigen via MHC class II, positive regulation of T cell activation, transcription from RNA polymerase II promoter, positive regulation of transcription, and DNA templated. While in CC, the downregulated DEGs were enriched in integral components of the lumenal side of the endoplasmic reticulum membrane, trans-Golgi network membrane, and in MF, peptide antigen binding, transcription factor binding, transcriptional activator activity, RNA polymerase II core promoter proximal region sequence-specific binding, MHC class II receptor activity, and MHC class I protein binding.Pathway enrichment analysis is frequently used to understand high throughput molecular data and produce hypotheses about the underlying biological processes of studies. The downregulated DEGs were associated with the T cell receptor signalling pathway, the intestinal immune network for IgA production, antigen processing and presentation, HTLV-I infection, toxoplasmosis, and the Epstein-Barr virus infection, according to functional enrichment analysis of integrated DEGs .ALB, GRAP2, HLA-DPA1, GATA3, HLA-DQB1, HLA-DPB1, SERPINE1, LEF1, PBX1, PHGDH, ICOS, KCNN4, ALDH1A1, NFATC2, and CFD) as shown in Figures PPI network analysis methods are an effective way to quicken our understanding of the biochemical and molecular interactions that underlie pathogenesis. A PPI network was built using the STRING online database , which wp = 0.012). Besides, the OS was lower in the low CFD expression group than that in the high expression group (p = 0.028). The other hub genes were not shown to be significantly linked to AML prognosis due to p > 0.05 , which are PDB ID: 7JWW [The molecular docking analysis is used to simulate the interaction of atomic level between a small molecule and a protein, allowing us to characterize how small molecules behave at the binding site of target proteins and enabling a better understanding of biological processes. Docking is a method for finding a suitable ligand that fits into the sites of a protein, energetically and geometrically. In other words, it is the study of how two or more molecules, such as ligands and proteins, interact with each other. The ligands can drive the functional changes of the target molecules, which are determined by their binding in the active sites of the targets , 21. BasID: 7JWW  for ALDHID: 7JWW  for CFD.Docking studies were carried out to investigate the molecular binding pattern of the compounds within the active pocket of protein surfaces. Tables In the interaction of ALDH1A1, the enasidenib formed carbon-hydrogen bonds with GLY458 and GLY294 residues  is used for analyzing the physical movements of atoms and molecules. Among the drug candidates, enasidenib and gilteritinib were chosen for dynamic simulation analysis after a virtual screening. To understand the dynamic activity of the protein-ligand complex in a solvent environment over time, MD simulations were carried out on the YASARA structure tool v. 20.12.24.W.64 (using the AMBER14 force field) with 100 ns.Additionally, the SASA or solvent-accessible surface areas of the four complexes was evaluated to see if the protein surface or volume had changed. In this study, the total SASA was computed which denotes the biomolecular surface area is accessible to solvent molecules. A higher SASA profile is associated with a longer protein volume, whereas a lower SASA profile is associated with a shorter protein volume. The SASA profile for both complexes had a higher profile and exhibited a stable profile during simulation The root mean square fluctuation (RMSF) plot  revealedBesides, as mentioned previously, the calculation of MM-PBSA binding energy for two complexes was done. CFD is upregulated, and ALDH1A1 is downregulated. The GEPIA tool was also used to validate the expression of the two genes. Such data validation confirmed the relation between the selected biomarkers and AML prognosis.AML is the most common in adults, which is a grievous, life-threatening, and often remediable hematological malignancy that influences the progenies of myeloid cells and individuals of all ages. Even though the availability of numerous treatments, such as chemotherapy, allogeneic hematopoietic stem cell transplantation , and receptor-antagonist medications, the 5-year survival of the patient is less than 30% . As a reSTAB1) is a prognostic factor of AML that were validated with three other independent CN-AML datasets [CALCRL, was used for determining the chemotherapy schedule and risk of HSCT in patients with AML/ETO+AML [ERCC3 was considered as a biomarker as its overexpressed elevated ERCC3 expression in AML patients [HSPA8 high expression was seen in another study in AML, and it was recognized as a possible independent prognostic factor in CN-AML patients [STAT1, BATF, and EML4 were identified as independent indicators of pediatric AML [MMP7 or MMP15 expression have a substantially poor prognosis, and this finding raises the possibility that MMP7 and MMP15 are potential prognostic markers and therapeutic targets for AML [POBEC3G gene as a potential prognostic marker of AML [A similar study identified that the scavenger receptor stabilin-1  in CSCs from human melanoma, making this isozyme a potential therapeutic target in melanoma [ALDH1A1 and AML. Thus, further investigation is needed to verify the association between ALDH1A1 and AML and evaluate the gene as a target for AML treatment. Although the precise biological functions of the CFD have only been studied by molecular biology approaches [ALDH1A1 and CFD are related to AML. Since this result has been found by the computational analysis of two data sets only, it needs to be validated by techniques of molecular biology, such as RT-PCR and western blot, using clinical samples. cancer. –44 and h cancer. , 62. Thi cancer. . ALDH1A1melanoma . Accordiproaches , 31, thiMolecular docking has been performed to find out the most effective drugs for AML from the list approved by the FDA. The first prediction of molecular recognition between ligand and target is heavily influenced by molecular docking. Molecular docking reveals the interacting amino acid residues, docking energy analysis, hydrogen bonding, and analysis of amino acid residues of the active sites and potential binding sites, which were used to decipher drug-target interactions . Variousin silico research [ALDH1A1 activity. The research showed that the protein backbone RMSD (1.55 Å) revealed that the structure was stabilized after 3,000 ps of simulation. Another study [The best compound was selected by MD simulation and free energy calculation on the top-scoring compounds Tables  and 5. Dresearch , the eller study  screeneder study  investigALDH1A1 and CFD, that are strongly associated with the progression of tumors and cancer. This suggests that the two biomarkers are likely to serve as prognostic indicators and therapeutic targets for AML. They had great docking energy with the FDA-approved AML drugs, gilteritinib and enasidenib, respectively. Moreover, the MD simulation validated the stability of their binding between active pockets of the proteins and compounds. This study also ensured the stages and clinical diagnosis of AML and, finally, provided the specific medication for a specific patient. The in silico results presented in this article need to be validated by laboratory research, which would be our future target. That is why, the authors are interested to confirm the function of these screened genes and pathways in the development of AML through future study by laboratory research (in vitro and in vivo).In summary, our investigation found 238 DEGs by integrating the two GEO datasets of AML. The PPI networks and survival analysis identified two important genes,


Ca2+ in the cytosol comprises a versatile signal controlling neuronal cell physiology to accommodate adaptive structural and functional changes of neuronal networks  and, as such, is essential for brain function. Although disease risk factors selectively affect different neuronal cell types across age-related neurodegenerative diseases (NDDs), these appear to have in common the ability to impair the specificity of the Ca2+ signal. As a result, non-specific Ca2+ signalling facilitates the development of intraneuronal pathophysiology shared by age-related NDDs, including mitochondrial dysfunction, elevated reactive oxygen species (ROS) levels, impaired proteostasis, and decreased axonal transport, leading to even more Ca2+ dyshomeostasis. These core pathophysiological processes and elevated cytosolic Ca2+ levels comprise a self-enforcing feedforward cycle inevitably spiralling toward high levels of cytosolic Ca2+. The resultant elevated cytosolic Ca2+ levels ultimately gear otherwise physiological effector pathways underlying plasticity toward neuronal demise. Ageing impacts mitochondrial function indiscriminately of the neuronal cell type and, therefore, contributes to the feedforward cycle of pathophysiology development seen in all age-related NDDs. From this perspective, therapeutic interventions to safely restore Ca2+ homeostasis would mitigate the excessive activation of neuronal destruction pathways and, therefore, are expected to have promising neuroprotective potential.This review postulates that age-related neurodegeneration entails inappropriate activation of intrinsic pathways to enable brain plasticity through deregulated calcium (Ca Age-related neurodegenerative diseases (NDDs) have in common the dysfunction and demise of neurons in the brain, particularly in the elderly population. Depending on the type and, therefore, function of the neurons subjected to degeneration, corresponding symptoms develop. For instance, in amyotrophic lateral sclerosis (ALS), motor neurons degenerate, leading to movement abnormalities, whereas in Alzheimer’s disease (AD), neurons operating in networks for memory formation and cognition deteriorate. In some cases, the underlying cause (or at least the most prominent cause) of age-related NDDs is known, such as in familial forms of neurodegeneration. A classic example entails familial Alzheimer’s disease (fAD), in which mutations in certain risk genes result in elevated production of neurotoxic amyloid beta (Aβ). However, far more often, the cause or causes leading to neurodegeneration in individual patients are not known. Nevertheless, many risk factors have been identified that increase the likelihood of developing neurodegeneration. These risk factors entail a plethora of conditions, characteristics, or lifestyles and can be either positive or negative (protective). Typically, these risk factors have a low penetrance and, therefore, in isolation, are not sufficient to cause neurodegeneration. Rather, it appears that the accumulation of several positive risk factors and/or the absence of negative risk factors at some point overwhelm neurons’ ability to maintain cellular homeostasis for optimal function . For exaA salient and poorly understood topic in the field is that, although typically all neurons of the CNS are exposed to, experience, or have experienced the same risk factors, typically, a selective population of neurons is the most vulnerable; hence, the different symptoms across NDDs. For instance, in the case of the fAD gene mutations mentioned above, hippocampal and cortical neurons are among the most vulnerable neurons, even though, obviously, all neurons carry these mutations. This is similar to ALS, in which specific risk gene mutations selectively affect motor neurons. Another notable observation is that although NDD risk factors are very diverse in nature and can impact different neuronal cell types, the intraneuronal pathophysiology in the affected neurons is remarkably similar, and neurodegeneration is the common outcome.Hence, important conceptual questions about the mechanisms of neurodegeneration are unanswered or poorly understood. What molecular mechanisms enable or execute neurodegeneration? What are the common elements of such mechanisms across different neurodegenerative diseases? What is the role of ageing? What underlies selective neuronal vulnerability? Addressing these questions will increase our understanding of mechanisms underlying age-related neurodegeneration and possibly facilitate drug discovery and development approaches to counter neurodegeneration in patients. This brief review aims to provide new perspectives in a rather conceptual manner to progress our thinking about these important questions.Neurodegeneration is not a passive phenomenon that occurs spontaneously under certain pathological or adverse conditions; it entails active processes and pathways geared towards the elimination of neurons and/or neuronal structures  to impact the physiology and function of the cell. Classic examples are receptor-mediated influx of Ca2+ through the inositol trisphosphate receptor from the endoplasmic reticulum (ER) or through the N-methyl-D-aspartate (NMDA) receptor at the plasma membrane upon stimulation by glutamate. Further, the kinetics of Ca2+ rise in the cytosol is another critical aspect of calcium control. Transient high increases in the cytosol may have different, and sometimes opposite, outcomes compared to prolonged modest elevations. For instance, whereas high transient increase potentiates synaptic strength, the opposite effect is caused by lower, more chronic elevations of cytosolic Ca2+ [In neurons (like in many other cell types), cytosolic Caed above . A key flic Ca2+ . In othelic Ca2+ .2+ signal, consequently leading to abnormal activity of otherwise physiological Ca2+ effectors, as outlined in 2+ concentration gradient , the Ca2+ signal in the cytosol is not properly specified, leading to non-selective and excessive activation of neuronal destruction pathways , extensive calcium transport, and organellar Ca2+ storage systems. It is, therefore, also a fragile system sensitive to adverse conditions. CALMH1 P86L polymorphism is an example of a genetic risk factor of sAD, which, through its Ca2+ activity, may directly impact Ca2+ homeostasis in AD. Other examples include mutations in presenilin 1 and -2 [2+ homeostasis. One important consequence of ageing is the progressive decline of mitochondrial function caused by reactive oxygen species (ROS), a byproduct of electron transfer chain activity. In turn, decreased mitochondrial function limits the production of ATP required for maintaining the Ca2+ gradient [2+ and act as important buffers to maintain Ca2+ homeostasis. Under conditions of excessive levels of calcium in the cytosol, the consequential excessive accumulation of intra-mitochondrial Ca2+ levels facilitates elevated ROS production, hence further contributing to ROS levels in the cell [2+ transporter activities necessary to maintain the Ca2+ gradient [2+ buffering capacity of mitochondria may become overwhelmed, triggering increased opening frequency of the mPTP pore followed by a release of pro-death signals such as cytochrome C [2+-sensitive destruction pathways, as listed in Maintaining the large Ca1 and -2  and the 1 and -2 . Ageing gradient ,69. Furtthe cell ,71. Furtgradient . At some2+ dyshomeostasis are intertwined, reinforcing processes [The above-outlined scenario illustrates a feedforward loop in that mitochondrial dysfunction, ROS production, and Carocesses ,71, whicrocesses . This, irocesses .2+ dyshomeostasis has been proposed [2+ concentration gradient [2+ influx by potentiating NMDA receptor activity [2+ in neurodegeneration is illustrated by the observations that impaired calcium homeostasis in itself facilitates the formation of tau and Aβ pathology [In the case of AD, the most prominent age-related NDD, a central role of Caproposed ,74,75,76gradient ,77. Likegradient ,79,80. Aactivity ,82. In Aathology ,83. In oathology .2+ dyshomeostasis and, consequently, neurodegeneration . In other words, what is the weakest link affected by adverse conditions? For instance, specific cellular or physiological determinants attributed to the neuronal identity confer selective vulnerability to alpha-synuclein or tau toxicity . In the f neuron ,95. If, f neuron ,97. Altef neuron , possiblf neuron .2+ dyshomeostasis, ER stress) underlies the common intraneuronal pathophysiology across NDDs.Irrespective of the above, ageing is a universal risk factor for age-related neurodegenerative disorders. In other words, it seems likely that ageing impacts pathophysiological elements shared by age-related NDDs. As outlined above, mitochondrial dysfunction is a prominent mechanism of biological ageing, suggesting the pathophysiology directly or indirectly associated with it , indicating that mitochondrial dysfunction and associated pathophysiology comprise common mechanisms underlying age-related neurodegeneration.Although disease risk factors and the correspondingly affected neuronal cell types differ across neurodegenerative diseases, surprisingly, the intraneuronal pathophysiology in the diseased neurons is remarkably similar. Prominent common “core” elements of intraneuronal pathophysiologies entail mitochondrial dysfunction, increased oxidative stress, calcium dyshomeostasis, reduced axonal trafficking, and impaired proteostasis ,100,101.2+ dyshomeostasis in a vicious feedforward cycle. Mitochondrial dysfunction with associated ROS production, impaired proteostasis, or calcium-induced calcium release (CICR) are prominent examples in which the cause and consequence of Ca2+ is dyshomeostasis. This is not surprising given the central role of Ca2+ in virtually all aspects of cell physiology, including those that control Ca2+ homeostasis. However, although the pathophysiology has common core elements, it appears that the neuropathology is (at least to some extent) disease-specific. For instance, the formation of Aβ pathology in cortical neurons in AD does not seem to occur in motor neurons in the case of ALS. Alternatively, aggregation of huntingtin is only observed in HD. Apparently, the specific outcome of shared pathophysiological processes, such as impaired or limited proteostasis, can result in a neuronal cell type and, thus, disease-specific protein deposition pathology , it ultimately leads to similar or overlapping pathophysiology across neurodegenerative diseases. An inevitable and common outcome constitutes a suboptimal or even a catastrophic rise of cytosolic calcium, executing neurodegeneration through the excessive activation of intrinsic mechanisms of neuronal demise (Collectively, risk factors of neurodegeneration may promote one or more elements of the shared pathophysiology . Given tl demise . Ageing,2+, leading to neurodegeneration. From this follows that therapeutic interventions that mitigate or prevent the pathological rise of cytosolic Ca2+ are expected to have a large therapeutic potential across different NDDs. After all, such an approach would break most effectively the anticipated vicious cycles of core pathophysiology processes leading to neuronal death. In other words, such interventions would enable neurons to be more resilient to adverse disease risk factors.The mechanism outlined in 2+ homeostasis safely are challenging precisely because Ca2+ has such a central role in regulating physiology, not only in neurons but in virtually all cells of the body. Targeting specific modulators of Ca2+ homeostasis (2+ homeostasis may be too limited to achieve a clinically meaningful lowering effect on cytosolic calcium because calcium dyshomeostasis is controlled by a multitude of different processes. An excellent example illustrating these points is memantine, one of the therapeutic options in AD (2+ dyshomeostasis in AD. Hence, although conceptually it illustrates that restoring Ca2+ homeostasis is a valid approach to treating AD, at the same time, these limitations seem to limit the effect size in patients.From a more practical perspective, however, approaches to normalising Caeostasis , such asns in AD . This dr2+-sensitive phosphatase that extensively regulates neuronal functionality and survival. In animal models of AD, CN inhibition restores synaptic plasticity with a large effect size [At present, no clinically validated therapeutic concepts are known that effectively normalise calcium homeostasis in diseased neurons of patients in a safe way. Some concepts or candidate drugs under study are listed in ect size ,120. In ect size . These d2+ effector pathways geared toward neuronal destruction of vulnerable neurons. Consequently, normalising Ca2+ homeostasis would be the most effective therapeutic intervention, provided it does not overly impact physiological calcium signalling.Collectively, this review outlines a mechanism of neurodegeneration in which risk factors of neurodegeneration impact directly or indirectly one or more core physiological processes in vulnerable neuronal populations predisposed to these risk factors: mitochondrial dysfunction, elevated ROS production, impaired proteostasis, decreased axonal trafficking, and calcium dyshomeostasis. Ageing is a risk factor that contributes to thepathophysiology indiscriminately of neuronal cell type. These pathophysiological processes are engaged with cytosolic calcium in a vicious feedforward cycle, which ultimately leads to excessive activation of Ca