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The influence of laminar flow on the corrosion of pure zinc (Zn) and cell response was investigated and compared with the static measurements. The results revealed that laminar flow accelerated the corrosion and enhanced the localized corrosion of pure Zn. The dynamic corrosion rate was 0.184 mm/yr after 168 h corrosion. More corrosion products were formed under flow condition, which were mainly composed of Zn3(PO4)2·4H2O, Ca3(PO4)2, ZnO and Zn5(OH)6(CO3)2. Moreover, the adhered rat aortic endothelial cells (RAEC) on Zn samples exhibited notably cell loss and round configuration under laminar flow.

Purpose To characterize a 10-year series of patients with mesial temporal lobe epilepsy (MTLE) and unilateral hippocampal sclerosis (HS) and determine the histopathological characteristic of the association between granule cell dispersion (GCD) and hippocampal neuronal loss. Methods The study included 108 MTLE/HS patients. Histopathological analyses were performed in NeuN-stained hippocampal sections for HS pattern, neuronal density, dentate gyrus (DG) pathology, and granule cell layer width. Statistical tests investigated the association between DG pathologies and HS patterns, as well as the correlation of DG width with total hippocampal and subfield-specific neuronal densities. Results Fifty-six patients (51.9%) presented right HS. All the four ILAE HS patterns were represented (90 Type 1, 11 Type 2, 2 Type 3, and 5 no-HS). Sixty-seven patients (62.0%) presented GCD, 39 (36.1%) normal DG, and 2 (1.9%) narrow DG. GCD was associated with initial precipitating injury, higher numbers of monthly focal seizures and lifetime bilateral tonic-clonic seizures, longer epilepsy duration, and older age at surgery. GCD was prevalent in all HS patterns, except for Type 2 (81.8% normal versus 18.2% GCD, p = 0.005). GCD was associated with total hippocampal and subfield-specific neuronal loss, except for CA1. DG width correlated with total hippocampal (r = -0.201, p = 0.037) and CA4 neuronal densities (r = -0.299, p = 0.002). Patients with HS Type 1 had better surgical outcomes, with 51 (61.4%) seizure-free in the first year post-surgery. Conclusions This study confirmed that seizure control in MTLE/HS patients submitted to surgical treatment is comparable worldwide. Moreover, histopathological analyses showed an association between GCD and hippocampal neuronal loss, especially in the CA4 subfield.

Total nucleated cell (TNC) and CD34+ cell doses are considered among the most important parameters when assessing the suitability of a human leukocyte antigen-matched cord blood unit (CBU) for allogeneic hematopoietic stem cell transplantation (HSCT). Cord blood banks therefore frequently select CBUs for cryopreservation based on pre-process TNC content. However, cell loss during processing can lead to a significant quantity of CBUs that do not meet desired post-process quality criteria, and such grafts are less likely to be selected by transplant centers for HSCT. Here the authors present a multi-parameter linear regression (MLR) model capable of identifying CBUs that would process poorly, despite meeting established pre-process TNC and CD34+ quality thresholds.Historically processed CBUs were graded from A+ to D depending on post-process cell content, and the utilization rate of each grade category was examined. Eight pre-process predictors of post-process cell content were used to train the MLR model, including red blood cell (RBC) content; CBU volume; age of CBU when received; and TNC constituent cell subsets. The selection efficacy of this model was then compared to that of methods conventionally used to select CBUs for processing, with receiver operating characteristic (ROC) and mean inventory quality analysis forming the basis of assessment.Within the Anthony Nolan Cell Therapy Centre, CBUs graded 'D' accounted for 37% of processing expenditures despite providing only 11% of grafts shipped for HSCT. The MLR model significantly improved pre-process identification of 'D' grade CBUs relative to thresholds based primarily on CD34+ cell content (P0.0001) and TNC content (P0.0001). At a comparable financial investment, this translated to a banked graft inventory of significantly higher quality than that produced by CD34+ (+8.8% mean increase, P = 0.007) and TNC (+9.9% mean increase, P = 0.010) selection methods.A predictive modelling approach to pre-process CBU selection is a simple and effective means to increase graft inventory quality and potentially future graft utilization, at no additional financial investment.

Lipid peroxidation (LPO) is the molecular mechanism involved in oxidative damage of cellular membranes and the hallmark of a nonapoptotic form of cell death, known as ferroptosis. This iron-dependent cell death is an emerging strategy in cancer treatment and one of the central cell death mechanisms accounting for early cell loss and organ dysfunction in both neurodegenerative disease and ischemia-reperfusion injury. Although the biological roles of LPO products have attracted considerable attention, not only for their pathological mechanisms but also for their potential clinical application as biomarkers, the existence of a common lethal lipid death signal generated during ferroptosis remains poorly explored. A better understanding of the LPO process, however, may unleash unprecedented opportunities for therapeutic intervention of as-yet incurable diseases.

Podocyte loss is a key element underlying glomerulosclerosis. Various mechanisms have been proposed for cell loss. These include local hemodynamic effects and local stresses on cell and hemodynamic changes, which together may contribute to detachment of podocytes, leading to sclerosis. Elegant studies based on classic observations add state-of-the-art imaging, modeling, intravital microscopy, and ultrastructural geometry analyses and provide new insights into potential mechanisms for injury and loss of this key cell.

Microwells are used in studies to mimic the in vivo environment through an in vitro environment by generating three-dimensional cell spheroids. These microwells have been fabricated in various shapes using different methods according to the research purpose. However, because all microwells up to now have an open top, it has been difficult to culture spheroids of floating cells due to their low density, such as human adipose-derived stem cells (hASCs) that differentiate into adipocytes. Therefore, the labor-intensive hanging droplet method has been mainly used for the study of adipocytes. Here, we introduce a sigma-well, which is a microwell in the shape of the Greek letter sigma (σ) with a roof. Because of its unique shape, the sigma-well is advantageous for the culture of floating cells by reducing cell loss and external interference. The sigma-well was fabricated using the principle of surface tension of polydimethylsiloxane as well as air trapping and thermal expansion. Unlike conventional microwells, because the center of the bottom surface and the inlet of the sigma-well are not located on the same line and have a difference of approximately 218 μm, the spheroids are cultured more stably and may not escape the cavity. In this study, hASC and adipocyte spheroids differentiated using these sigma-wells were successfully cultured. In addition, through cytokine diffusion simulation, it was confirmed that the diffusion and mass transfer in the sigma-well was lower than that in the conventional microwell. It is expected that the morphological features of the sigma-well, which cannot be easily obtained by other methods, can be beneficial for the study of buoyant cell types such as adipocytes.

Objective Adaptive changes in DHEA and sulfated-DHEA (DHEAS) production from adrenal zona reticularis (ZR) have been observed in normal and pathological conditions. Here we used three different cohorts to assess timing differences in DHEAS blood level changes and characterize the relationship between early blood DHEAS reduction and cell number changes in women ZR. Materials and methods DHEAS plasma samples (n = 463) were analyzed in 166 healthy prepubertal girls before pubarche ( Results We confirmed that overweight girls exhibited higher and earlier DHEAS levels and no difference was found compared with the average European and South American girls with a similar body mass index (BMI). Adrenopause onset threshold (AOT) defined as DHEAS blood levels 35% of the females >40 years old and associated with significantly reduced ZR cell number (based on PTEN and hematoxylin signals). ZR cell loss may in part account for lower DHEA/DHEAS expression, but most cells remain alive with lower DHEA/DHEAS biosynthesis. Conclusion The timely relation between significant reduction of blood DHEAS levels and decreased ZR cell number at the beginning of the 40s suggests that adrenopause is an additional burden for a significant number of middle-aged women, and may become an emergent problem associated with further sex steroids reduction during the menopausal transition.

Adeno Associated Virus (AAV)-mediated gene expression in the brain is widely applied in the preclinical setting to investigate the therapeutic potential of specific molecular targets, characterize various cellular functions, and model central nervous system (CNS) diseases. In therapeutic applications in the clinical setting, gene therapy offers several advantages over traditional pharmacological based therapies, including the ability to directly manipulate disease mechanisms, selectively target disease-afflicted regions, and achieve long-term therapeutic protein expression in the absence of repeated administration of pharmacological agents. Next to the gold-standard iodixanol-based AAV vector production, we recently published a protocol for AAV production based on chloroform-precipitation, which allows for fast in-house production of small quantities of AAV vector without the need for specialized equipment. To validate our recent protocol, we present here a direct side-by-side comparison between vectors produced with either method in a series of in vitro and in vivo assays with a focus on transgene expression, cell loss, and neuroinflammatory responses in the brain. We do not find differences in transduction efficiency nor in any other parameter in our in vivo and in vitro panel of assessment. These results suggest that our novel protocol enables most standardly equipped laboratories to produce small batches of high quality and high titer AAV vectors for their experimental needs.

Background: There are not many studies conducted in India to compare cell block preparation methods with reagents and materials that are readily available in all laboratories. This study aimed to standardize and compare two simple cell block techniques, which can be done in low resource settings too. In the study, 35 cases of thyroid, lymph node, and breast were collected for both FNA and cell block preparation for six months. Materials and Methods: There were separate passes given for both methods. A total of seventy cell blocks made using formalin and agar methods of preparation. Results: We compared both the methods on technical and morphological levels. The formalin method was overall easy to perform and was yielding good morphological results in 98% cases, the only drawback being cell loss during handling and processing. While in the agar method, there was almost no cell loss, but it was more technically difficult and yielded poorer morphological results. A scoring system was made for cellularity: no cells = 0, hypo-cellular = 1+, hypo-cellular with tissue fragments = 2+, cellular = 3+.18 A score of 2+ and 3+ was scored by 31/35 formalin blocks and 28/35 agar blocks. Conclusions: The sensitivity of both formalin and agar methods are almost comparable. However, the procedure of the formalin method is far more straightforward and user friendly. Moreover, it also provides a better architectural picture than the agar method.

Hepatitis D virus (HDV) requires hepatitis B surface antigen (HBsAg) for its assembly and release. Current HBV treatments are only marginally effective against HDV because they fail to inhibit HBsAg production/secretion. However, monotherapy with the nucleic acid polymer REP 2139-Ca is accompanied by rapid declines in both HBsAg and HDV RNA. We used mathematical modeling to estimate HDV-HBsAg-host parameters and to elucidate the mode of action and efficacy of REP 2139-Ca against HDV in 12 treatment-naive HBV/HDV co-infected patients. The model accurately reproduced the observed decline of HBsAg and HDV, which was simultaneous. Median serum HBsAg half-life (t1/2) was estimated as 1.3 [0.9–1.8] days corresponding to a pretreatment production and clearance of ~108 [107.7–108.3] IU/day. The HDV-infected cell loss was estimated to be 0.052 [0.035–0.074] days−1 corresponding to an infected cell t1/2 = 13.3 days. The efficacy of blocking HBsAg and HDV production were 98.2 [94.5–99.9]% and 99.7 [96.0–99.8]%, respectively. In conclusion, both HBsAg production and HDV replication are effectively inhibited by REP 2139-Ca. Modeling HBsAg kinetics during REP 2139-Ca monotherapy indicates a short HBsAg half-life (1.3 days) suggesting a rapid turnover of HBsAg in HBV/HDV co-infection.

Hybridomas are a commonly used, or even the only option, for laboratory study and pilot production of monoclonal antibodies (mAbs), which are crucial for both targeted therapy and biomedical study. A long-term culture of hybridomas will inevitably induce a heterogenization of the whole hybridoma population, resulting in a continuous growth of non-producing hybridomas. To overcome the limits of existing methods of screening heterogeneous hybridomas, in which the whole multi-round screening process is performed in multi-well plates or other discrete modules, this study presents a novel method in which all processing steps of a multi-round hybridoma screening are finished in a single microfluidic chip. This microfluidic chip comprehensively performs hybridoma trapping/proliferating/transferring and fluorescent identification of protein-antibody binding at single cell level. By performing a two-round screening of anti-CD45 mAb secreting hybridomas, the novel microfluidic chip was proved capable of screening several single high-producing hybridomas with minimum cell loss/human labor/time cost, and more importantly, enhanced accuracy and definite monoclonality, which is one of the most important properties of mAb production.

Killing pathogens by different electrochemical (EC) disinfection means has been largely reported in the literature, even if the influence of process variables and reactor conception on kill performance has not been well comprehended. This review concentrates on EC microbial killing mechanisms especially the free radicals’ contribution and the effect of the electric field (EF), which are by their nature poisonous to microbes. Some mechanisms have been suggested to interpret the deadliness of EC application. Such pathways comprise: 1) oxidative stress and cell loss of life because of electrochemically produced oxidants, 2) irreversible permeabilization of cell membranes by the placed EF, 3) electrooxidation of vital cellular constituents during exposure to electric current or induced EFs, and 4) electrosorption of negatively charged E. coli cells to the anode surface followed by direct electron transfer reaction. Future investigations have to be more dedicated to the EF influence in the EC disinfection, as it is the main part of the involved mechanisms. Employing granular activated carbon post-treatment could greatly reduce the concentrations and poisonous effects of disinfection by-products. Moreover, secure multi-barrier techniques, like distillation, plasma discharge, nanotechnologies, and membrane processes remain to be suggested, tested, and industrially encouraged. Despite their limitations, both adsorptive techniques and membrane processes persist to be an encouraging domain of research thanks to their relatively low costs and ease of applications.

Purpose: To analyze if 6-month endothelial cell density (ECD) affects long-term ECD outcome and graft survival 5 years after Descemet membrane endothelial keratoplasty (DMEK) in eyes with Fuchs endothelial corneal dystrophy (FECD).Design: Retrospective cohort study.Participants: A total of 585 DMEK eyes were included. The study group was divided into 4 groups based on 6-month ECD quartiles: group 1 (n = 146) with 313 to 1245 cells/mm(2), group 2 (n = 148) with 1246 to 1610 cells/mm(2), group 3 (n = 145) with 1611 to 1938 cells/mm(2), and group 4 (n = 146) with 1939 to 2760 cells/mm(2). Group 1 was further split into subgroups 1a (n = 36) with 6-month ECD of

PURPOSE: To use empirical data to develop a model of cell loss in choroideremia that predicts the known exponential rate of RPE loss and central, scalloped preservation pattern seen in this disease. METHODS: A computational model of RPE loss was created in Python 3.7, which constructed an array of RPE cells clusters, binarized as either live or atrophic. Two rules were applied to this model: the background effect gave each cell a chance of dying defined by a background function, and the neighbor effect increased the chance of RPE cell death if a neighbor were dead. The known anatomic distribution of rods, RPE, choriocapillaris density, amacrine, ganglion, and cone cells were derived from the literature and applied to this model. Atrophy growth rates were measured over arbitrary time units and fit to the known exponential decay model. The main outcome measures: included topography of atrophy over time and fit of simulated residual RPE area to exponential decay. RESULTS: A background effect alone can simulate exponential decay, but does not simulate the central island preservation seen in choroideremia. An additive neighbor effect alone does not simulate exponential decay. When the neighbor effect multiplies the background effect using the rod density function, our model follows an exponential decay, similar to previous observations. Also, our model predicts a residual island of RPE that resembles the topographic distribution of residual RPE seen in choroideremia. CONCLUSIONS: The pattern of RPE loss in choroideremia can be predicted by applying simple rules. The RPE preservation pattern typically seen in choroideremia may be related to the underlying pattern of rod density. Further studies are needed to validate these findings.

Neurodegenerative diseases are characterized by selective vulnerability of distinct cell populations; however, the cause for this specificity remains elusive. Many circuits that degenerate in disease are shaped by neural activity during development, raising the possibility that mechanisms governing early cell loss may be misused when activity is compromised in the mature brain. Here we show that electrical activity and synaptic transmission are both required for neuronal survival in the adult entorhinal cortex, but these silencing methods trigger distinct means of degeneration in the same neuronal population. Competition between active and inactive cells drives axonal disintegration caused by synaptic inhibition, but not axon retraction due to electrical suppression. These findings suggest that activity-dependence may persist in some areas of the adult brain long after developmental critical periods have closed. We speculate that lifelong plasticity required to support memory may render entorhinal neurons vulnerable to prolonged activity changes in disease.

Pathologic complete response (pCR) predicts the long-term outcome of neoadjuvantly treated (NAC) breast cancer (BC) but is reached in &lt, 10% of hormone-receptor-positive patients. Biomarkers enabling adjustment or interruption of an ineffective therapy are desired. Here, we evaluated whether changes in the serum concentration of thymidine kinase 1 (sTK1) during NAC could be utilized as a biomarker. In the PROMIX trial, women with localized HER2- BC received neoadjuvant epirubicin/docetaxel in six cycles. sTK1 was measured with an ELISA in 54 patients at cycles 1–4 and in an additional 77 patients before and 48 h after treatment 1. Treatment resulted in a 2-fold increase of sTK1 before and a 3-fold increase 48 h after the cycles, except for the first cycle, where half of the patients reacted with a significant decrease and the other half with an increase of sTK1. In Kaplan–Meier estimates of ER+ patients divided by the median of the post/pre-treatment sTK1 ratio at the first treatment cycle, OS was 97.7% and 78% (p = 0.005), and DFS was 90.7% and 68% (p = 0.006), respectively. Thus, the response of sTK1 at the first cycle of chemotherapy could be used both as an early biomarker for the guidance of chemotherapy and for the study of inherent tumor chemo-sensitivity, which could predict long-term outcome prior to therapy.

This paper concerns a free boundary problem modeling tumor growth with angiogenesis and two time delays. The two delays represent the time taken for cells to undergo mitosis and modify the rate of cell loss because of apoptosis, respectively. We study the stability of stationary solutions and find that Hopf bifurcation occurs under some conditions, which extends the results of Xu. Furthermore, numerical simulations are performed to investigate the relationship among the rate of angiogenesis, two time delays and Hopf bifurcation., 14 pages, 6 figures

The clinical use of doxorubicin (DOX) is largely limited by its cardiotoxicity. Previous studies have shown that jaceosidin has many biological activities. However, little is known about whether jaceosidin can attenuate DOX-related acute cardiotoxicity. Here, we investigated the therapeutic effects of jaceosidin on DOX-induced acute cardiotoxicity. Mice were intraperitoneally injected with a single dose of DOX to establish an acute cardiac injury model. To explore the protective effects, mice were orally administered jaceosidin daily for 7 days, with dosing beginning 2 days before DOX injection. The results demonstrated that jaceosidin dose-dependently reduced free radical generation, inflammation accumulation, and cell loss induced by DOX in cardiomyocytes. Further studies showed that jaceosidin treatment inhibited myocardial oxidative damage and the inflammatory response and attenuated myocardial apoptotic death, thus improving cardiac function in mice injected with DOX. The inhibitory effects of jaceosidin on DOX-related acute cardiotoxicity were mediated by activation of the sirtuin1 (Sirt1) signaling pathway. Jaceosidin lost its protective effect against DOX-related injury in Sirt1-deficient cardiomyocytes and mice. In conclusion, jaceosidin has protective potential in treating DOX-related cardiac injury through activation of the Sirt1 signaling pathway.

The gold standard animal model of Parkinson’s disease is the non-human primate rendered parkinsonian with the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) toxin. Low availability, ethical issues, and primate-specific biohazards make alternative large animal models necessary. Here, we investigate the temporal evolution of presynaptic dopaminergic function after MPTP in another large animal model, the Göttingen minipig. We subcutaneously injected seven sedated minipigs with 1–2 mg/kg of MPTP, and two minipigs with saline, three times a week over 4 weeks. We monitored behavioral deficits using a validated motor scale and a Gait4Dog® walking mat. Minipig brains were imaged with (+)-⍺-[11C]-dihydrotetrabenazine ([11C]-DTBZ) and [18F]-fluorodopa ([18F]-FDOPA) PET at baseline and 1, 3, 9 and 12 months after the final MPTP injection. Immunohistochemical tyrosine hydroxylase (TH) staining was used to assay nigral TH + area loss post-mortem. The minipigs showed only mild bradykinesia and impaired coordination at early timepoints after MPTP. PET revealed decreases of striatal [11C]-DTBZ and [18F]-FDOPA uptake post-MPTP with a partial spontaneous recovery of [18F]-FDOPA after 9 months. Postmortem histological analysis showed a loss of 71% TH-immunopositive area in the substantia nigra. When testing the efficacy of putative neuroprotective agents, partial spontaneous recovery of dopamine terminal function must be taken into account in the MPTP minipig model of parkinsonism.

Water-in-oil emulsion droplet microfluidic systems have been extensively developed, and currently, almost all cell handling steps can be conducted in this format. An exception is the cell washing and solution exchange step, which is commonly utilized in many conventional cell assays. This paper presents an in-droplet cell washing and solution exchange technology that utilizes dielectrophoretic (DEP) force to move all cells to one side of a droplet, followed by asymmetrical splitting of the droplet to obtain a small daughter droplet that contains all or most of the cells, and then finally merges this cell-concentrated droplet with a new droplet that contains the desired solution. These sequential droplet manipulation steps were integrated into a single platform, where up to 88% of the original solution in the droplet could be exchanged with the new solution while keeping cell loss to less than 5%. Two application examples were demonstrated using the developed technology. In the first example, green microalga Chlamydomonas reinhardtii cells were manipulated using negative DEP force to exchange the regular culture medium with a nitrogen-limited medium to induce lipid production. In the second example, Salmonella enterica cells were manipulated using positive DEP force to replace fluorescent dye that models fluorescent cell stains that contribute to high background noise in fluorescence-based droplet content detection with fresh buffer solution, significantly improving the droplet content detection sensitivity. Since the cell washing step is one of the most frequently utilized steps in many cell biology assays, we expect that the developed technology can significantly broaden the type of assay that can be conducted in droplet microfluidic format.

Summary Neuronal intranuclear inclusion disease (NIID) is a neurodegenerative disease characterized by the presence of intranuclear inclusions of unknown origin. NIID is caused by an expansion of GGC repeats in the 5′ UTR of the NOTCH2NLC (N2C) gene. We found that these repeats are embedded in a small upstream open reading frame (uORF) (uN2C), resulting in their translation into a polyglycine-containing protein, uN2CpolyG. This protein accumulates in intranuclear inclusions in cell and mouse models and in tissue samples of individuals with NIID. Furthermore, expression of uN2CpolyG in mice leads to locomotor alterations, neuronal cell loss, and premature death of the animals. These results suggest that translation of expanded GGC repeats into a novel and pathogenic polyglycine-containing protein underlies the presence of intranuclear inclusions and neurodegeneration in NIID., Graphical abstract, Highlights • NIID is a neurodegenerative disease caused by expansion of GGC repeats in NOTCH2NLC • These GGC repeats are translated into a polyglycine (polyG) protein • The polyG protein is toxic and forms intranuclear inclusions in cells and animals • Similarities between FXTAS and NIID define a new set of disorders: polyG diseases, The neurodegenerative disease NIID is caused by an expansion of GGC repeats in NOTCH2NLC. Boivin et al. found that these repeats are translated into a toxic polyglycine (polyG) protein that forms intranuclear inclusions. An identical mechanism exists in FXTAS, unveiling a novel group of genetic pathologies, the polyG diseases.