Presence Of Apoptotic Cells Research Articles

Effect of estrogen depression on alveolar bone microarchitecture and periodontal ligament cells during orthodontic movement.

This study aimed to evaluate the effects of the estrogen depression during orthodontic tooth movement on alveolar bone microarchitecture and periodontal ligament. Female Wistar rats were divided into two groups, one consisting of non-ovariectomized animals subjected to orthodontic tooth movement, and one comprising ovariectomized animals subjected to orthodontic tooth movement. Micro-CT assessment of bone volume to total volume (BV/TV), total porosity, trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular separation (Tb.Sp) in the alveolar bone of the orthodontically moved tooth was performed. Histomorphometric analyses were made in the periodontal ligament, and immunoexpression of RANK, RANKL, OPG, and TUNEL were quantified. Orthodontic tooth movement in the group of ovariectomized rats was faster than in non-ovariectomized animals. The alveolar bone area showed lower values of BV/TV and trabecular thickness, and higher bone porosity and trabeculae numbers in the ovariectomized rats. Histological analyses in the ovariectomized group revealed an increase in collagen fibers in the periodontal ligament. The apoptotic cell counts in the periodontal ligament were higher in the group of ovariectomized rats than in the sham-operated rats. Ovariectomy resulted in an increase in tooth movement and alteration of the alveolar bone microstructure in the first 7day of orthodontic tooth movement, and in the presence of apoptotic cells in the periodontal ligament.

Oral lichen planus – an oral potentially malignant disorder (OPMD) of the oral cavity

Introduction. Oral lichen planus is a chronic inflammatory disease of unknown etiology, characterized by recurrent lesions, presenting as reticular lesions, sometimes accompanied by atrophic, erosive, and/or ulcerative areas. Despite being one of the most common conditions affecting the oral mucosa, oral lichen planus remains an ailment with undefined etiology and unclear pathogenesis, imprecise management, and uncertain premalignant potential. Materials and methods. A narrative literature review study was conducted. A bibliographic search was carried out in databases such as PubMed, Hinari, SpringerLink, the National Center of Biotechnology Information, and Medline. Articles published from 1990 to 2023 were selected using various combinations of keywords: “oral lichen planus,” and “epidemiology,” “etiology,” “pathogenesis,” “symptoms,” “management,” “histopathology,” and “malignant transformation.” After processing the data from these databases, 475 full articles were found. The final bibliography comprised 50 relevant sources, considered representative of the materials published on the topic of this synthesis article. Results. Oral lichen planus is an inflammatory condition associated with T-cell-mediated immune dysfunction. Triggers include autoimmune responses to local antigens, microorganisms, and stress. The disease results from a complex interplay of host factors, lifestyle, and environmental factors leading to T-cell-mediated immune dysregulation. Diagnosis of oral lichen planus is based on clinical features (multiple, bilateral, symmetrically distributed lesions, occurring most commonly on the buccal mucosa, dorsal tongue surface, and gingiva), histopathological findings (predominantly lymphocytic bandlike infiltrate in the lamina propria, presence of apoptotic cells in the basal cell layer, absence of epithelial dysplasia), and immune-related changes (deposition of fibrinogen along the basement membrane zone, presence of granular fluorescent deposits containing IgA, IgG, and IgM in colloid bodies). Conclusions. Oral lichen planus is a chronic inflammatory condition mediated by T-cells in response to various extrinsic antigens, modified autoantigens, or superantigens, with periods of remission and relapse and the potential for malignant transformation. The etiology and pathogenesis of this condition are complex, diagnosis relies on clinical features, histopathological findings, and immunological data, patient treatment is symptomatic, and the potential for malignant transformation varies. Nevertheless, prospective studies with large sample sizes, adequate treatment duration, and long-term follow-up are needed.

A Novel Drosophila Model of Alzheimer's Disease to Study Aβ Proteotoxicity in the Digestive Tract.

Amyloid-β (Aβ) proteotoxicity is associated with Alzheimer's disease (AD) and is caused by protein aggregation, resulting in neuronal damage in the brain. In the search for novel treatments, Drosophila melanogaster has been extensively used to screen for anti-Aβ proteotoxic agents in studies where toxic Aβ peptides are expressed in the fly brain. Since drug molecules often are administered orally there is a risk that they fail to reach the brain, due to their inability to cross the brain barrier. To circumvent this problem, we have designed a novel Drosophila model that expresses the Aβ peptides in the digestive tract. In addition, a built-in apoptotic sensor provides a fluorescent signal from the green fluorescent protein as a response to caspase activity. We found that expressing different variants of Aβ1-42 resulted in proteotoxic phenotypes such as reduced longevity, aggregate deposition, and the presence of apoptotic cells. Taken together, this gut-based Aβ-expressing fly model can be used to study the mechanisms behind Aβ proteotoxicity and to identify different substances that can modify Aβ proteotoxicity.

Non-monotonic effects of Bisphenol A Dimethacrylate on male mouse reproductive system and fertility leads to impaired conceptive performance

As an estrogenic agent, Bisphenol A Dimethacrylate (Bis-DMA) may incite alterations in both the reproductive tract and the neuroendocrine axis, and thus have the potential to affect the proper development, maturity and conceptive performance in animals. We investigated the consequences of 14 weeks of exposure to different concentrations of Bis-DMA on male mouse conceptive performance. Male mice were exposed to Bis-DMA (0, 0.1 mg/L, 1.0 mg/L or 10 mg/L) via drinking water, and the effects on fertility, reproductive organ weights, reproductive hormone levels, sperm counts and testicular histology were assessed. We clearly demonstrate that prolonged exposure of male mice to Bis-DMA negatively affects fertility and reproduction causing significant reductions in sperm counts, non-monotonic effects on serum LH and testosterone levels, increased seminal vesicle weights, lower number of embryonic implantations and viable fetuses, as well as, increased embryonal resorptions in females mated by Bis-DMA treated males. Furthermore, Bis-DMA caused abnormalities in testicular infrastructure with atrophic seminiferous tubules exhibiting intraepithelial vacuolization and disorganization, loss and shedding of germ cells into the lumen, and presence of apoptotic cells. Our data collectively suggest that Bis-DMA adversely affects male fertility and reproduction by interference with normal hormone signaling in the testis, inducing changes in testicular infrastructure and ultimately leading to impaired reproductive function and fertility.

Effectiveness of Gold Nanorods of Different Sizes in Photothermal Therapy to Eliminate Melanoma and Glioblastoma Cells.

Gold nanorods are the most commonly used nanoparticles in photothermal therapy for cancer treatment due to their high efficiency in converting light into heat. This study aimed to investigate the efficacy of gold nanorods of different sizes (large and small) in eliminating two types of cancer cell: melanoma and glioblastoma cells. After establishing the optimal concentration of nanoparticles and determining the appropriate time and power of laser irradiation, photothermal therapy was applied to melanoma and glioblastoma cells, resulting in the highly efficient elimination of both cell types. The efficiency of the PTT was evaluated using several methods, including biochemical analysis, fluorescence microscopy, and flow cytometry. The dehydrogenase activity, as well as calcein-propidium iodide and Annexin V staining, were employed to determine the cell viability and the type of cell death triggered by the PTT. The melanoma cells exhibited greater resistance to photothermal therapy, but this resistance was overcome by irradiating cells at physiological temperatures. Our findings revealed that the predominant cell-death pathway activated by the photothermal therapy mediated by gold nanorods was apoptosis. This is advantageous as the presence of apoptotic cells can stimulate antitumoral immunity in vivo. Considering the high efficacy of these gold nanorods in photothermal therapy, large nanoparticles could be useful for biofunctionalization purposes. Large nanorods offer a greater surface area for attaching biomolecules, thereby promoting high sensitivity and specificity in recognizing target cancer cells. Additionally, large nanoparticles could also be beneficial for theranostic applications, involving both therapy and diagnosis, due to their superior detection sensitivity.

Transitory alkali exposure on meibomian gland orifices induces meibomian gland dysfunction

PurposeTo determine pathological changes of meibomian glands (MGs) after transient exposure of the rat eyelid margin to alkali solution. MethodsFilter paper infiltrated with 1 N sodium hydroxide solution was applied to the eyelid margin of Sprague-Dawley rats for 30 s under general anesthesia, without touching the conjunctiva, after which the ocular surface and eyelid margin were examined by slit-lamp microscopy. In vivo confocal microscopy and stereomicroscopy were subsequently applied to observe MG morphology on day 5, day 10 and day 30 post alkali injury. Eyelid cross-sections were processed for H&E staining, Oil red O staining and immunofluorescent staining. ResultsAfter alkali injury, there was marked plugging of MG orifices, telangiectasia and hypertrophy of the eyelid margin, while corneal epithelium was intact at post-injury days 5 and 10. However, 30 days after alkali injury, mild corneal epithelial damage was observed. Degeneration of MG acini was observed at days 5 and became aggravated at days 10 and 30, along with MG duct dilation and acini loss. Oil red O staining showed lipid accumulation in the dilated duct. Inflammatory cell infiltration and the presence of apoptotic cells was seen in the MG loci 5 days post injury, but diminished at days 10 and 30. Cytokeratin 10 expression was increased in dilated duct, while cytokeratin 14, PPAR-γ, Ki67 and LRIG1 expression were decreased in the acini of injured loci. ConclusionsTransitory alkali exposure of the rat eyelid margin obstructs the MG orifice and induces pathological changes of MG dysfunction.

Functional Blockage of S100A8/A9 Ameliorates Ischemia-Reperfusion Injury in the Lung.

(1) Background: Lung ischemia-reperfusion (IR) injury increases the mortality and morbidity of patients undergoing lung transplantation. The objective of this study was to identify the key initiator of lung IR injury and to evaluate pharmacological therapeutic approaches using a functional inhibitor against the identified molecule. (2) Methods: Using a mouse hilar clamp model, the combination of RNA sequencing and histological investigations revealed that neutrophil-derived S100A8/A9 plays a central role in inflammatory reactions during lung IR injury. Mice were assigned to sham and IR groups with or without the injection of anti-S100A8/A9 neutralizing monoclonal antibody (mAb). (3) Results: Anti-S100A8/A9 mAb treatment significantly attenuated plasma S100A8/A9 levels compared with control IgG. As evaluated by oxygenation capacity and neutrophil infiltration, the antibody treatment dramatically ameliorated the IR injury. The gene expression levels of cytokines and chemokines induced by IR injury were significantly reduced by the neutralizing antibody. Furthermore, the antibody treatment significantly reduced TUNEL-positive cells, indicating the presence of apoptotic cells. (4) Conclusions: We identified S100A8/A9 as a novel therapeutic target against lung IR injury.

P848: TINOSTAMUSTINE (EDO-S101), AN ALKYLATOR AND HISTONE DEACETYLASE INHIBITOR, ENHANCES THE EFFICACY OF DARATUMUMAB IN PRECLINICAL MODELS OF MULTIPLE MYELOMA

Background: Tinostamustine (EDO-S101) is a novel alkylating and deacetylase inhibiting molecule designed to improve drug access to DNA strands, induce DNA damage and counteract its repair in cancer cells. It has shown anti-myeloma (MM) activity in different preclinical models both in monotherapy and in combination with bortezomib. Aims: To evaluate whether tinostamustine enhances the anti-myeloma effect of daratumumab. Methods: Tinostamustine was provided by Mundipharma and daratumumab was obtained from pharmacy surplus of the University Hospital of Salamanca. Mechanisms of action of daratumumab were studied by flow cytometry (FCM) in MM cell lines pretreated with tinostamustine and cultured as specified: 1) with F(ab)2 fragments (direct apoptosis via crosslinking); 2) with 10% human serum (CDC assays); 3) co-cultures of MM and NK cells (ADCC assays). Expression of CD38 and NKG2D ligands (MICA and MICB) after tinostamustine treatment was evaluated by FCM, WB and qPCR. The effects of tinostamustine + daratumumab combination were also studied ex vivo in bone marrow (BM) samples from MM patients, and in vivo in two plasmacytoma models developed in CB17-SCID and NK-cell-humanized NSG mice. Results: Pretreatment of U266, JJN3, MM.1S, NCI-H929, RPMI-8226 and MOLP-8 cell lines with tinostamustine (1-2.5 μM) for 48 h increased CD38 expression. Tinostamustine-pretreated myeloma cells showed a higher level of daratumumab binding, as demonstrated by anti-human-IgG1-AF488 staining. Also, tinostamustine (1-2.5 μM) increased MICA and MICB expression in MM cell lines. In ex vivo cultures, tinostamustine increased expression of CD38 in 4 out of 10 patients and MICA in 3 out of 5 patients. Next, the influence of tinostamustine pretreatment (2.5 μM) on several daratumumab mechanisms was evaluated. In apoptosis via crosslinking, the percentage of apoptotic cells in presence of daratumumab was higher in tinostamustine-pretreated MOLP-8 cells vsDMSO-pretreated cells (90.57%±7.35 vs 60.36%±2.86; p<0.001). Likewise, pretreatment with tinostamustine increased the percentage of Annexin-V+/7AAD+ MOLP-8 cells in CDC assays with daratumumab, although not reaching statistical significance (93.22%±1.05 vs73.39%±11.93). In MM-NK co-cultures in absence of daratumumab (NK cell-mediated direct cytotoxicity), the percentages of apoptotic cells in tinostamustine-pretreated vs DMSO-pretreated cells were: 83.64%±5.28 vs 61.58%±6.81 in MOLP-8 cells (p<0.05); 54.27%±5.99 vs 34.24%±7.37 for RPMI-8226 (p<0.05); and 26.15%±4 vs 12.32%±1.97 in MM.1S (p<0.05). However, in the presence of daratumumab (ADCC) these percentages showed a similar tendency although not significant: 88.57%±3.77 vs 78.89%±5.57 in MOLP-8; 70.75%±5.69 vs 58.28%±7.52 for RPMI-8226; and 56.74%±7.25 vs 45.68%±7.1 in MM.1S. Ex vivo experiments with patients’ BM samples (n = 10) showed that simultaneous combination of daratumumab + tinostamustine significantly increased the percentage of eliminated MM cells compared to individual treatments, with an acceptable toxicity on healthy lymphocytes (Figure 1a). Finally, in the CB17-SCID model, tinostamustine treatment (24 h) followed by daratumumab administration controlled tumor growth significantly better than daratumumab alone (Figure 1b). The combination prolonged the median survival as compared to vehicle, and monotherapy with tinostamustine or daratumumab (Figure 1b). Similar results were obtained in the NSG model. Image:Summary/Conclusion: Our preclinical data demonstrate that tinostamustine increases the anti-myeloma effect of daratumumab presumably due to enhanced expression of CD38 and MICA.

Intravitreal Administration of rhNGF Enhances Regenerative Processes in a Zebrafish Model of Retinal Degeneration.

Nerve growth factor (NGF) is the best characterized neurotrophin, and it is known to play an important role in ocular homeostasis. Here, we demonstrated the expression of NGF receptors in adult zebrafish retina and optimized a light-induced retina degeneration (LID) zebrafish model that mimics human cone-rod disorders, demonstrating that intravitreal (IV) administration of rhNGF can boost zebrafish retinal regeneration in this model. Adult zebrafish retinae exposed to 60 h of light irradiation (60 h LID) displayed evident reduction of outer nuclear layer (ONL) thickness and cell number with presence of apoptotic cells. Retinal histologic evaluation at different time points showed that IV therapeutic injection of rhNGF resulted in an increase of ONL thickness and cell number at late time points after damage (14 and 21 days post injury), ultimately accelerating retinal tissue recovery by driving retinal cell proliferation. At a molecular level, rhNGF activated the ERK1/2 pathway and enhanced the regenerative potential of Müller glia gfap- and vim-expressing cells by stimulating at early time points the expression of the photoreceptor regeneration factor Drgal1-L2. Our results demonstrate the highly conserved nature of NGF canonical pathway in zebrafish and thus support the use of zebrafish models for testing new compounds with potential retinal regenerative properties. Moreover, the pro-regenerative effects of IV-injected NGF that we observed pave the way to further studies aimed at evaluating its effects also in mammals, in order to expedite the development of novel rhNGF-based therapeutic approaches for ophthalmological disorders.

Bfc, a novel serpent co-factor for the expression of croquemort, regulates efferocytosis in Drosophila melanogaster.

Efferocytosis is the process by which phagocytes recognize, engulf, and digest (or clear) apoptotic cells during development. Impaired efferocytosis is associated with developmental defects and autoimmune diseases. In Drosophila melanogaster, recognition of apoptotic cells requires phagocyte surface receptors, including the scavenger receptor CD36-related protein, Croquemort (Crq, encoded by crq). In fact, Crq expression is upregulated in the presence of apoptotic cells, as well as in response to excessive apoptosis. Here, we identified a novel gene bfc (booster for croquemort), which plays a role in efferocytosis, specifically the regulation of the crq expression. We found that Bfc protein interacts with the zinc finger domain of the GATA transcription factor Serpent (Srp), to enhance its direct binding to the crq promoter; thus, they function together in regulating crq expression and efferocytosis. Overall, we show that Bfc serves as a Srp co-factor to upregulate the transcription of the crq encoded receptor, and consequently boosts macrophage efferocytosis in response to excessive apoptosis. Therefore, this study clarifies how phagocytes integrate apoptotic cell signals to mediate efferocytosis.

Nuclear PKR in retinal neurons in the early stage of diabetic retinopathy in streptozotocin‑induced diabetic rats.

Retinal neuron apoptosis is a key component of diabetic retinopathy (DR), one of the most common complications of diabetes. Stress due to persistent hyperglycaemia and corresponding glucotoxicity represents one of the primary pathogenic mechanisms of diabetes and its complications. Apoptosis of retinal neurons serves a critical role in the pathogenesis of DR observed in patients with diabetes and streptozotocin (STZ)-induced diabetic rats. Retinal neuron apoptosis occurs one month after STZ injection, which is considered the early stage of DR. The molecular mechanism involved in the suppression of retinal neuron apoptosis during the early stage of DR remains unclear. RNA-dependent protein kinase (PKR) is a stress-sensitive pro-apoptotic kinase. Our previous study indicated that PKR-associated protein X, a stress-sensitive activator of PKR, is upregulated in the early stage of STZ-induced diabetes. In order to assess the role of PKR in DR prior to apoptosis of retinal neurons, immunofluorescence and western blotting were performed to investigate the cellular localization and expression of PKR in the retina in the early stage of STZ-induced diabetes in rats. PKR activity was indirectly assessed by expression levels of phosphorylated eukaryotic translation initiation factor 2α (p-eIF2-α) and the presence of apoptotic cells in the retina was investigated by TUNEL assay. The findings revealed that PKR was localized in the nucleus of retinal ganglion and inner nuclear layer cells from normal and diabetic rats. To the best of our knowledge, the present study is the first to demonstrate nuclear localization of PKR in retinal neurons. Immunofluorescence analysis demonstrated that PKR was expressed in the nuclei of retinal neurons at 3 and 6 days and its expression was decreased at 15 days after STZ treatment. In addition, p-eIF2-α expression and cellular localization followed the trend of PKR, suggesting that this pro-apoptotic kinase was active in the nuclei of retinal neurons. These findings are consistent with the hypothesis that nuclear translocation of PKR may be a mechanism to sequester active PKR, thus preventing upregulation of cytosolic signalling pathways that induce apoptosis in retinal neurons. Apoptotic cells were not detected in the retina in the early stage of DR. A model was proposed to explain the mechanism by which apoptosis of retinal neurons by PKR is suppressed in the early stage of DR. The possible role of mitochondrial RNA (mtRNA) and Alu RNA in this phenomenon is also discussed since it was demonstrated that the cellular stress due to prolonged hyperglycaemia induces the release of mtRNA and transcription of Alu RNA. Moreover, it mtRNA activates PKR, whereas Alu RNA inhibits the activation of this protein kinase.

Mechanical analysis of oriented cell movement by nanostructure-based biosensors

Due to the numerous diseases caused by apoptosis, the existing methods for detecting early apoptosis are not effective. To explore the effect of nanostructured biosensors on the directional movement of cells, silica@quantum dot-concanavalin A (SiO2@QDs-ConA) nanoprobes were prepared, and cholesterol amperometric biosensors based on carbon nanotubes were constructed by using the layer-by-layer assembly method. The results showed that in the presence of apoptotic cells, a sharp anodic stripping voltammetric signal was obtained, with a peak potential of about −0.7 V, corresponding to the oxidation peak of the cadmium (Cd2+) ion of the combined quantum dot nanoprobe. The signal of apoptotic cells was 3.9 and 15.0 times higher than those of normal cells and no cells, respectively, indicating the detection of apoptotic cells by electrochemical cell sensors in the early stage. Eight hours before the action of the apoptosis-inducing reagent, part of HL-60 cells underwent apoptosis. Sixteen hours after the action of the reagent, the apoptosis of HL-60 cells was obvious, the action time of the apoptosis-inducing reagent increased, the peak current of cadmium ion enrichment and dissolution increased rapidly and the HL-60 cells had obviously undergone apoptosis, indicating that most of the cells had obviously undergone apoptosis at this time, indicating that use of the constructed sensor is feasible for the early detection of apoptotic cells.

Apoptosis in epididymis of sand rat Psammomys obesus, Cretzschmar, 1828: Effects of seasonal variations, castration and efferent duct ligation

The aim of this study was to visualize apoptosis throughout the reproductive cycle and after castration, castration then treatment with testosterone, and ligation of efferent ducts. The sand rat, Psammomysobesus, Cretzschmar 1828, is a diurnal rodent belonging to the family Gerbillidae. Its breeding cycle is seasonal with reproduction in autumn, winter and early spring and a short resting period from late spring to early summer. Five groups of males were studied: (1) animals captured during the breeding season; (2) animals captured during the resting season; (3) animals castrated and kept 30 days; (4) animals castrated, kept 30 days, and then treated with testosterone for 15 days; (5) animals subjected to the ligation of efferent ducts and kept 30 days. Epididymis were removed and the presence of apoptotic cells was explored using the "Apostain" immunohistochemical method. Histological results showed cell and tissue remodeling. During the breeding season, a positive apoptotic signal was observed mainly in smooth muscle cells of caput and cauda epididymis. This signal persisted throughout the resting season. The orchiectomy induced apoptosis in almost of epithelial and connective cells. However, this intense cell death was not reversed by treatment with testosterone. In animals that experienced efferent duct ligation, principal cells and smooth muscle cells showed a positive signal for apoptosis. Our results converge to qualify the sand rat epididymis as an excellent model for the study of apoptosis and argue for continued cell death, at least independent of circulating testosterone levels.

Anagallis arvensis Induces Apoptosis in HL-60 Cells Through ROS-Mediated Mitochondrial Pathway

The present study was taken up to evaluate the apoptosis inducing ability of alcoholic extract of whole plant of Anagallis arvensis (AAE) in HL-60 cells. We observed time and concentration dependent decrease in cell viability after treatment with AAE. Fluorescent staining and scanning electron micrographs of treated HL-60 cells demonstrated chromatin condensation, nuclear fragmentation and formation of apoptotic blebs. There was a marked increase in hypodiploid population of cells as observed by cell cycle analysis. Annexin V-FITC/PI also depicted the presence of apoptotic cells. Anti-apoptotic protein Bcl-2 was observed to be decreased by 62% at 20 µg/ml concentration and a significant increase in ROS production up to 6.9-fold was observed in time dependent manner. In addition, alteration in mitochondrial membrane potential was observed, which was followed by cytochrome c release to cytoplasm. Activated levels of mitochondrial downstream pathway protein namely Caspase-3 and 9, were detected in treated HL-60 cells by colorimetric analysis. DNA ladder formation, a biochemical hallmark of apoptosis was also observed in treated HL-60 cells. The results of the present study support the apoptotic potential of AAE and probability of its promising role in development as effective anticancer agent against leukemia cells.

Microglia dynamics in retinitis pigmentosa model: formation of fundus whitening and autofluorescence as an indicator of activity of retinal degeneration

In patients with retinitis pigmentosa (RP), color fundus photography and fundus autofluorescence (FAF) have been used to estimate the disease progression. To understand the origin and the diagnostic interpretation of the fundus color and FAF, we performed in vivo imaging of fundus color and FAF together with histological analyses of the retinal degeneration process using the RP model mice, rd10. FAF partly represented the accumulation of microglia in the photoreceptor outer segments. Fundus whitening suggested the presence of apoptotic cells, which spatiotemporally preceded increase in FAF. We observed two patterns of FAF localization, arcuate and diffuse, each indicating different pattern of apoptosis, wavy and diffuse, respectively. Diffuse pattern of apoptosis was suppressed in dark-raised rd10 mice, in which outer nuclear layer (ONL) loss was significantly suppressed. The occupancy of FAF correlated with the thinning rate of the ONL. Fractalkine, a microglia chemotactic factor, was detected in apoptotic photoreceptors, suggesting chemokine-induced recruitment of microglia into the ONL, which paralleled with accelerated ONL loss and increased FAF occupancy. Thus, we propose that the degree of photoreceptor apoptosis and the rate of ONL thinning in RP patients might be read from the fundus color and the FAF.

Overfeeding-Induced Obesity Could Cause Potential Immuno-Physiological Disorders in Rainbow Trout (Oncorhynchus mykiss).

Simple SummaryIn this study, we have successfully generated overfeeding-induced obesity in rainbow trout and demonstrated that the overfeeding regime causes adverse effects on the health status of fish. What we found in this study is that fish in the overfed group harbor enlarged liver and macrophages due to high lipid accumulation. The expression of IL-10, CD36, TLR2, and HSP70 (stress and/or obesity-related genes) was significantly upregulated in overfed fish. Moreover, oxidized low-density lipoprotein (OxLDL), which is known to be produced more in obese individuals, caused apoptosis of trout lymphocyte. These results clearly indicate that overfeeding-induced obesity can be the source of stress and cause immuno-physiological disorders in rainbow trout. Awareness and knowledge of farmers on the relationship between overfeeding and fish health might contribute to increased disease resistance and aquaculture production.Although over-nutrition from overfeeding-induced obesity is known to be highly associated with metabolic and immunological disorders in humans, little is known about overfeeding-induced obesity in fish farming. The purpose of this study was to investigate changes in immuno-physiological parameters, to better understand the potential risk of overfeeding–induced obesity in fish. Commercial feed was provided to fish in the overfed group until they refuse to eat, but fish in the control group was fed with the feed at 1% bodyweight per day. The hemato-serological, histological, and immunological changes were observed at weeks 2 and 8. Rainbow trout leukocytes were co-incubated with oxidized low-density lipoprotein (OxLDL), and the phagocytes engulfing the OxLDL and the presence of apoptotic cells were evaluated. The body weight, body mass index (BMI), and hepatosomatic index (HSI) index were significantly higher in the overfed group, and high lipid accumulation and fatty changes were also observed in their livers, indicating that the feeding regime used in this study led to overfeeding-induced obesity. Likewise, much higher numbers of and larger vacuoles were observed in overfed fish macrophages, showing unclear boundaries between the cytoplasm and extracellular space. In the overfed group, the expression of IL-10, HSP70, TLR2, and CD36 was significantly higher, and lymphocyte apoptosis was more evident, indicating that overfeeding-induced obese fish might have immunologic disorders. This was the first study to demonstrate that overfeeding-induced obesity could cause an immune-physiological imbalance in rainbow trout, making them more vulnerable to infectious diseases and various stressful conditions. This study will contribute to improvements in fish nutrition, feeding practices, fish nutrition, and disease prevention in the aquaculture industry.

Aptamer (AS1411)-Conjugated Liposome for Enhanced Therapeutic Efficacy of miRNA-29b in Ovarian Cancer.

AS1411 Aptamer-functionalized liposome was successfully formulated and found to be nanosized. Flow cytometer and CLSM results demonstrated that Aptamer enhanced the targeting of carrier in the cancer cells via nucleolin-mediated transmembrane endocytosis pathway. The lipofectaminebased miR-29b showed a typical concentration-dependent cytotoxic effect in the cancer cells. LP-miR induced a significant reduction in the cell viability of A2780 cells compared to that of nontreated control, while LP-Mut (mutant loaded) did not have any effect on the cell viability indicating the importance of the specific gene sequencing. LP-miR induced a significant decrease in the green fluorescence which is indicative of the decrease in the cell viability. Simultaneously, higher PI positive cells were observed for LP-miR treated cancer cells in Live/Dead assay. Cells treated with LP-miR exhibited the brightest fluorescence indicating the presence of apoptotic cells. Significant increase in the Annexin-V+ cells and PI+ cells were observed for cell treated with LP-miR compared to that of non-treated control indicating the potential of miR-29b. This novel miR-29b-loaded Aptamer-directed liposome could potential serve as a new platform to improve the therapeutic outcome in ovarian cancers.

Enhanced anti-lung carcinoma and anti-biofilm activity of fungal molecules mediated biogenic zinc oxide nanoparticles conjugated with β-D-glucan from barley

This work reports the optimization, synthesis, characterization, anticancer, and antibacterial activity of the Trichoderma-β-D-glucan‑zinc oxide nanoparticles (T-β-D-glu-ZnO NPs). Firstly, the T-ZnO NPs was synthesized using the fungal mycellial water extract (FWME) derived from T. harzianum (SKCGW009) under the optimized condition of extract concentration (5.99 mL), temperature (43.11 °C), pH (8) and time (69.04 h). The successful conjugation of T-ZnO NPs with β-D-glucan (T-β-D-glu-ZnO NPs) was confirmed by PACE and FTIR. The XRD, UHR SEM, and TEM EDS results pointed the spherical shape of NPs with the mean size of 30.34 nm. Further, the XPS survey scan and high-resolution fitting of Zn2p results also claimed the successful formation of the T-β-D-glu-ZnO NPs. Cytotoxicity results indicated that the NPs were not toxic to NIH3T3 cells, while exhibited the dose-dependent inhibitory effect to human pulmonary carcinoma A549 cells. The IC50 of T-ZnO NPs and T-β-D-glu-ZnO NPs against A549 cells was 158 and 56.25 μg.mL−1, respectively, which was also verified by fluorescent cytochemistry. Annexin V-FITC staining results indicated the presence of apoptotic cells in the NPs treated A549 cells, which was not seen in the non-treated control A549 cells. Interestingly, the number of necrosis cells was higher in the T-ZnO NPs (3.38%) comparing to T-β-D-glu -ZnO NPs (0.07%). The early or late apoptosis was found higher in the cells treated T-β-D-glu -ZnO NPs (6.43%) comparing with T-ZnO NPs (4%). These results indicated that T-ZnO NPs and T-β-D-glu-ZnO NPs induced the cancer cell death through necrosis and apoptosis pathway, respectively. The antibacterial results indicated that the NPs treatment were significantly inhibited the growth of the Staphylococcus aureus inside of roundworm and enhanced growth of roundworm. Overall, anticancer and in vitro, in vivo antibacterial studies proved the high caliber of T-β-D-glu-ZnO NPs for the further pharmaceutical evaluation.

The Role of MicroRNA-21 in Venous Neointimal Hyperplasia: Implications for Targeting miR-21 for VNH Treatment.

The molecular mechanism of hemodialysis access arteriovenous fistula (AVF) failure due to venous neointimal hyperplasia (VNH) is not known. The role of microRNA-21 (miR-21) in VNH associated with AVF failure was investigated by performing in vivo and in vitro experiments. In situ hybridization results revealed that miR-21 expression increased and was associated with fibroblasts in failed AVFs from patients. In a murine AVF model, qRT-PCR gene expression results showed a significant increase in miR-21 and a decrease in miR-21 target genes in graft veins (GVs) compared to contralateral veins in mouse AVF. miR-21 knockdown in GVs was performed using a lentivirus-mediated small hairpin RNA (shRNA), and this improved AVF patency with a decrease in neointima compared to control GVs. Moreover, loss of miR-21 in GVs significantly decreased the Tgfβ1, Col-Ia, and Col-Iva genes. Immunohistochemistry demonstrated a significant decrease in myofibroblasts and proliferation with an increase in terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining in miR-21-knockdown vessels, along with a decrease in hypoxia-inducible factor-1 alpha (HIF-1α) and phospho-SMAD2 (pSMAD-2) and phospho-SMAD3 (pSMAD-3) and an increase in phosphatase and tensin homolog (PTEN) staining. Hypoxic fibroblast knockdown for miR-21 showed a significant decrease in Tgfβ-1 expression and pSMAD-2 and -3 levels and a decrease in myofibroblasts. These results indicate that miR-21 upregulation causes VNH formation by fibroblast-to-myofibroblast differentiation.

The morphological characteristics of the Meckel’s diverticulum of pre-hatch and post-hatch broiler chicken

Morphology of the Meckel’s diverticulum (MD) of broiler chicken was investigated using gross anatomical, histological and transmission electron microscopy techniques. Forty fertile eggs and 100 chicks of Marshall broiler chicken were used for the study. Samples were collected from 6 randomly selected embryos or chicks on embryonic days 14, 17 and 19, and post-hatch days 1, 3, 5, 7, 14, 21, 28, 35, 42 and 56. The results showed that the mean lengths of the small intestine from the gizzard to the MD and from the MD to the ileocecal junction increased steadily with age. On embryonic days 14, 17 and 19, the poorly differentiated tunics of the MD were composed of lamina epithelialis mucosae and indistinct lamina propria mucosae as well as submucosal areas. The tunica muscularis exhibited loose islands of nascent smooth muscle fibres. Whereas on post-hatch day 1, the lamina epithelialis mucosae showed scanty microvilli and enterocytes with large cytoplasmic vacuoles, the MD of 3-day-old post-hatch chicks contained lymphatic tissues in the lamina propria mucosae. On post-hatch day 35, the lamina epithelialis mucosae demonstrated the presence of apoptotic cells while numerous lymphocytes at various stages of differentiation were observed within the lamina propria mucosae and tunica submucosa. In conclusion, the present study has provided clear structural evidence which shows that intense cellular proliferation occurs in the MD of broiler chicken between embryonic days 14 and 19. This organ may play significant roles in immunological surveillance of the gut, and such functions may peak on post-hatch day 35.