Trypan Blue Dye Research Articles

Invitro germination and optimization of basal media for protocorm-like bodies proliferation in Dieniaophrydis(J. Koenig) Seidenf

Dienia ophrydis is an endangered terrestrial orchid used for its ornamental and therapeutic purposes. This orchid possesses an anti-malarial alkaloid known as Malaxin. There is ongoing exploitation in the wild due to illegitimate herbal collection and climate change. To control the potential risk of extinction, there are no conservational measures or in vitro protocols established in this genus. The study focuses on addressing a germination protocol and enhancement of protocorm-like bodies (PLB) proliferation by optimizing the basal media and vitamins. It also aims to improve the misinterpretation of tetrazolium viability results in orchid testa along with a permeability test. Further, the seeds were subjected to 1% TTC staining to perform embryo viability test and 0.4% trypan blue (TB) dye for testa permeability. Ten basal media such as Gamborg's medium, Mitra medium, Thomale GD medium, Vacin and Went medium, Knudson C medium, Knop's medium, MS medium and ½ MS medium, ¼ MS medium, and modified MS medium (MMS) were tested for seed germination. The Half-strength MS medium responded with 92% of the highest germination and T-GD medium is not suitable for seed germination. The seed viability and permeability result showed 93% of permeability, 72% viability, and 9% dead seeds. Protocorms were transferred to the modified mMS medium containing half-strength macronutrients, full micronutrients, and 10 ppm of thiamine in vitamin formulation. PLB proliferation was observed and 12 shoots emerged from a single protocorm explant. Subsequently, the seedlings were acclimatized with 8-0-24 ratios of nitrogen, phosphorus, and potassium (NPK) as determined by soil testing. The study contributes in experimenting with seed germination of terrestrial orchid seeds asymbiotically and to establishing a protocol for conservation programs, especially in this genus.

Intraoperative corneal staining during cataract surgery

Intraoperative corneal staining with trypan blue dye is an uncommon complication of cataract surgery that occurs at the time of staining of the anterior capsule. Inadvertent corneal staining with Descemet’s membrane detachment (DMD) occurred in a 70-year-old male, while stromal edema following corneal staining occurred in an 80-year-old female while attempting to stain the anterior capsule. DMD was managed with air tamponade, and stromal edema was conservatively managed. One must thus be aware of corneal staining with trypan blue during cataract surgery. This complication can be avoided by using sharp-tipped instruments, proper wound construction, and proper depth perception, which can be done by adjusting the microscope.

Peptide L13S Derived from the BRN2 POU Domain Reduces Metastasis In Vivo and Inhibits Cell Migration and Invasion In Vitro.

The Brain-Specific Homeobox/POU Domain Protein 2 (BRN2) transcription factor supports melanoma progression by regulating the expression of several genes involved in cell migration and invasion. We hypothesized that a peptide designed based on the POU domain of BRN2 could block the BRN2 transcription activity and, consequently, reduce metastasis. Cell viability was accessed by Trypan Blue exclusion dye assay and xCelligence platform. Wound-healing scratch assay and transwell invasion with matrigel membrane assay were performed to analyze cell migration and invasion. The internalization mechanism of the L13S peptide was investigated using confocal microscopy and wound-healing scratch assay. The impact of L13S on cell protein expression was analyzed through western blotting. In vivo assays were conducted to evaluate the protective effect and toxicity of L13S in a metastatic model using murine melanoma cells. Here, we show that the peptide named L13S can inhibit the migration and invasion of murine melanoma cells (B16F10-Nex2) as well as the migration of human melanoma cells (SK-MEL-25 and A375) by regulating the expression of proteins involved in motility. Mechanistically, we found that L13S is internalized by murine melanoma cells via macropinocytosis and binds actin filaments and nuclei. More importantly, in vivo studies indicated that the peptide was able to significantly inhibit lung metastasis in syngeneic models without off-target effects and with virtually no cytotoxicity toward normal organs. L13S peptide is a strong candidate for further development as an anticancer agent for the treatment of melanoma metastasis.

Evaluation of Antitumor Activity of Sivanar Amirtham – A Herbomineral Formulation belonging to Indian Traditional System of Medicine using Dalton's Ascites Lymphoma in Mice

The purpose of the present study is to evaluate the antitumor activity (ATA) of traditional herbal preparation Sivanar Amirtham (SA) on Dalton’s Lymphoma Ascites. Siddha medicine system (SMS) is a traditional system of medicine originated from ancient Tamilakam of South India. Siddha medicine is a traditional healing system from Tamilakam in ancient South India. For our purpose, we have performed acute toxicity (AT) study as per OECD guidelines 423 and ATA by xenograft method. In this study, a single dose of 300, 1000 and 2000 mg/kg of Sivanar Amirtham suspension (SAS) was orally (p.o.) administered in mice and animals were observed for 14 days. For antitumor study (ATS), we have used DAL cells which were intraperitoneally (i.p.) inoculateded into mouse. The ATAs were studied by monitoring the parameters such as cell growth inhibitors, tumor weight measurements, mean survival time of DAL bearing mice as well as changes in depleted haematological and biochemical parameters due to tumorigenesis. The SAS was also evaluated for in vitro cytotoxicity study in different concentration and the viability of cells was determined by exclusion method of trypan blue dye (TBD). The AT study showed no signs of toxicity and no mortality after single administration of SAS. SAS caused significant decrease in packed cell volume (PCV) (value), Tm volume (value) and viable cell count (value), and it prolonged the life span of DAL Tm carrying mice. Haematological and biochemical profiles were reverted to normal levels in SAS treated mice. The results of in vitro cytotoxicity show that SAS showed significant ATA in mice with moderate DAL levels. The IC50 value turned into discovered to be 800 μg/ml from the in vitro cytotoxicity examine. The study strongly suggests that SAS has the potential to be an antitumor medication against DAL cells induced Tm and it can be extrapolated for further cancer (CA) prevention applications.

Using Electroporation to Improve and Accelerate Zebrafish Embryo Toxicity Testing.

Zebrafish have emerged as a useful model for biomedical research and have been used in environmental toxicology studies. However, the presence of the chorion during the embryo stage limits cellular exposure to toxic elements and creates the possibility of a false-negative or reduced sensitivity in fish embryo toxicity testing (FET). This paper presents the use of electroporation as a technique to improve the delivery of toxic elements inside the chorion, increasing the exposure level of the toxins at an early embryo stage (<3 h post-fertilization). A custom-made electroporation device with the required electrical circuitry has been developed to position embryos between electrodes that provide electrical pulses to expedite the entry of molecules inside the chorion. The optimized parameters facilitate material entering into the chorion without affecting the survival rate of the embryos. The effectiveness of the electroporation system is demonstrated using Trypan blue dye and gold nanoparticles (AuNPs, 20-40 nm). Our results demonstrate the feasibility of controlling the concentration of dye and nanoparticles delivered inside the chorion by optimizing the electrical parameters, including pulse width, pulse number, and amplitude. Next, we tested silver nanoparticles (AgNPs, 10 nm), a commonly used toxin that can lower mortality, affect heart rate, and cause phenotypic defects. We found that electroporation of AgNPs reduces the exposure time required for toxicity testing from 4 days to hours. Electroporation for FET can provide rapid entry of potential toxins into zebrafish embryos, reducing the time required for toxicity testing and drug delivery experiments.

Effects of Imatinib and Artemisia vulgaris extracts in combination on leukemia cell proliferation

Resistance to imatinib develops gradually and presents major complications in patients with chronic myeloid leukemia (CML) disease, necessitating additional study for therapy improvement. Artemisia vulgaris is a member of the genus Artemisia which is traditionally used in herbal medicine. Historically, this plant was used to cure a variety of many human diseases including cancer. The goal of this study was to investigate the combined effect of imatinib and A. vulgaris extract on CML cell lines. For this objective, TCCY cells were treated for 72 hours with Imatinib, A. vulgaris extract, and the combinations at various concentrations. The proliferation of cells was determined by using the Trypan Blue dye exclusion assay. The combined efficacy of extract and Imatinib was determined using a nonlinear isobologram model at IC80 levels. To find discrepancies between observed and predicted data, Friedman tests and Nemenyi post-hoc analysis were used. To confirm the combined impact, the Wilcoxon signed-ranks test was utilized. The results suggested that the effect of Imatinib and A. vulgaris in combination was additive to synergistic. As a result, it was found that taking A. vulgaris extract can make Imatinib more effective against the CML cell line TCCY.

Phytochemical Profiling and Assessment of Habbe Musaffe Khoon Against Cervical Cancer

Background In India, cancer of the cervix is the second most common cancer in women. Nowadays, various traditional herbal formulations can be used as adjuncts during conventional cancer therapies due to their anticancer activity. Habbe Musaffi Khoon (HMK), a Unani compound formulation, is traditionally used as a blood purifier and thus used to treat boils, scabies, acne, and pimples. It is also indicated in skin diseases, Habis-ud-Dam (styptic), and therapeutically used in Nafz-ud-Dam (hemorrhagic). Objectives The main purpose of this work was to phytochemically characterize HMK aqueous extract (HMKaq) and determine its activity against cervical cancer cells. Materials and Methods HMKaq was subjected to liquid chromatography-mass spectrometry (LC-MS) and evaluated for the presence of phytocompounds. HMKaq was evaluated for its effect on the viability of cervical cancer cells (HeLa, SiHa, and C33A) and was determined by MTT assay. Trypan blue dye exclusion was used to determine the outcome of HMKaq on the growth kinetics of cervical cancer cells. The role of HMK in regulating the cell cycle was analyzed by FACS and its effect on apoptosis was evaluated by checking mitochondrial membrane potential readings of JC-1 aggregates on a microplate reader and on the mRNA expression of caspase-3 and cytochrome C was evaluated by PCR. Results LC-MS analysis revealed 3845 compounds, out of which 58 were the major compounds. These included phenolics, esters, fatty acids, furans, quinones, tannins, flavonoids, and terpenes. HMKaq reduced the viability and growth of the cells. It induced arrest in the cell cycle of HeLa and apoptosis in SiHa and C33A by depolarizing the membrane potential of the mitochondria and upregulation of mRNA expression of Cas3 and Cyt C. Conclusion HMK exhibited anticancer activity against cervical cancer cells, thereby signifying its therapeutic potential.

Management of an inadvertent late filtering bleb and hypotony in a scleral fixated intraocular lens implantation.

To report a case of inadvertent cystic filtering bleb in a patient with scleral fixated intraocular lens (SFIOL) implantation 7 years after initial procedure. A 14-year-old boy underwent lensectomy, vitrectomy and a sutured rigid SFIOL for microspherophakia in the right eye. Seven years later, the patient presented with hypotony related ocular changes. Examination revealed an inadvertent cystic filtering bleb at the scleral section. Anterior segment optical coherence tomography (ASOCT) revealed gap in inner lips of the scleral wound, bridged by a thin strip of scleral tissue. A conservative management with bandage contact lens (BCL) failed after an initial improvement. Injection of trypan blue dye in the anterior chamber confirmed the presence and site of wound filtration. Surgical closure of the fistula was done and patient showed both structural and visual improvement. Meticulous wound construction and water tight closure at the end of the surgery can avoid such complications. ASOCT can be helpful in diagnosing the area of fistula. A conservative management with BCL can help by tamponading the filtering bleb. However, in unresponsive cases, surgical closure of the fistula is recommended to improve the hypotony-related changes.

Biological activities of the aerial and undergound parts of Gymnadenia nigra Rchb.f. (syn. Nigritella nigra (L.) Rchb. f.) from the Italian Alps

This study investigated the bioactivity of both aerial (GNAR) and underground (GNUG) parts of Gymnadenia nigra Rchb.f. (syn. Nigritella nigra (L.) Rchb. f.) (Orchidaceae). The obtained data proved interesting when the samples were tested in two adrenocortical cancer cell lines (SW13 and H295R). In particular, the GNAR 80% methanol extract distinctly inhibited their viability after 24 h at a concentration of 1 µg/µL by MTT assay and trypan blue dye exclusion method. Cell morphology evaluation by means Wright’s staining also showed significant results, particularly in SW13 cells under the effect of both extracts. GNAR extract was able to scavenge the DPPH radical better than GNUG extract. It also was more active in albumin denaturation (a maximum % denaturation equal to 463.0 ± 8.3 vs 77.3 ± 13.3) and protease inhibition (a maximum % inhibition equal to 138.5 ± 7.0 vs 2.1 ± 2.0) tests. The results highlighted an important antitumor activity of G. nigra in vitro that deserves to be further studied.

Long-Term Cryopreservation of Peripheral Blood Stem Cell Harvest Using Low Concentration (4.35%) Dimethyl Sulfoxide with Methyl Cellulose and Uncontrolled Rate Freezing at -80 °C: An Effective Option in Resource-Limited Settings

Long-term cryopreservation of peripheral blood stem cells (PBSCs) is highly useful in the setting of tandem/multiple transplantations or treatment of relapse in the autologous hematopoietic stem cell transplantation (HSCT) setting. Even in allogeneic HSCT, donor lymphocyte infusions may be stored for months to years if excess stem cells are collected from donors. Cryopreservation is a delicate, complex, and costly procedure, and higher concentrations of dimethyl sulfoxide (DMSO), a commonly used cryoprotectant, can be toxic to cells and cause adverse effects in the recipient during infusions. In this study, we examined the effect of long-term cryopreservation using 4.35% DMSO (as final concentration) with methyl cellulose and uncontrolled rate freezing in a mechanical freezer (-80 °C) on the viability and colony-forming ability of CD34+ human PBSCs. For patients undergoing autologous HSCT, PBSCs were cryopreserved using DMSO (final concentration of 4.35%) with methyl cellulose. The post-thaw viability of PBSCs was determined using Trypan blue exclusion and flow cytometry-based 7-amino-actinomycin-D (FC-7AAD) methods. Concentrations of CD34+ stem cells and immune cell subsets in post-thaw PBSC harvest samples were assessed using multicolor flow cytometry, and the clonogenic potential of post-thaw stem cells was studied using a colony-forming unit (CFU) assay. CD34+ stem cell levels were correlated with the prestorage CD34 levels using the Pearson correlation test. The viability results in the Trypan blue dye exclusion method and the flow cytometry-based method were compared using Bland-Altman plots. We studied 26 PBSC harvest samples with a median cryopreservation duration of 6.6 years (range, 3.8 to 11.5 years). The median viability of post-thaw PBSCs was >80% using both methods, with a weak agreement between them (r=.03; P=.5). The median CD34+ stem cell count in the post-thaw samples was 9.13×106/kg (range, .44 to 26.27×106/kg). The CFU assay yielded a good proliferation and differentiation potential in post-thaw PBSCs, with a weak correlation between granulocyte macrophage CFU and CD34+ stem cell levels (r=.4; P=.05). Two samples that had been cryopreserved for >8 years showed low viability. Cryopreservation of PBSCs using 4.35% DMSO with methyl cellulose and uncontrolled freezing in a mechanical freezer at -80 °C allows the maintenance of long-term viability of PBSC for up to 8 years.

Biodegradation and decolorization of trypan blue azo dye by marine bacteria Vibrio sp. JM-17

Marine microorganisms have shown the capability of azo dye degradation that is used in the textile and tanning industries and cause harmful environmental effects. In the present study, we isolated Vibrio sp. from the marine environment which decolorizes trypan blue dye (direct blue 14). The decolorization by Vibrio sp. was 90.33% in 6 day at 500 mg L−1 of dye concentration. The study of environmental parameters showed Vibrio sp. as well adapted bacterial isolate in trypan blue degradation and capable of decreasing dye load in industrial effluent. The high-performance liquid chromatography (HPLC) results showed a complete conversion of the major peak into the two new peaks. The formation of new peaks was further confirmed by functional group (SO, C–N, C–O, OH group) analysis by Fourier-transform infrared (FTIR) spectroscopy. The gas chromatography-mass spectrometry (GC-MS) showed less harmful small molecules i.e., the formation of by-products (alkane, alkene, fatty acid and benzene) after trypan blue dye degradation by bacterial isolate. Enzyme assay showed major involvement of azoreductase and tyrosinase in the degradation of trypan blue dye. In the proposed pathway, the decrease in toxicity of direct blue- 14 dye by destabilization of the di-azo bond and formation of the less harmful by-product as benzoic acid. The toxicity analysis of dye-degraded by-products (DDP) at different concentrations (1–25%) carried on seeds of Vigna radiata showed 100% of seed germination. Similarly, the toxicity of DDP at a 5% concentration was studied on edible seaweed Ulva lactuca which showed an increase in daily growth rate (DGR) with an increase in content of both chlorophyll a and chlorophyll b after 10 days of incubation. The results of the present study showed that bacterial isolate Vibrio sp. is capable of degradation of trypan blue dye with an environmentally friendly approach.

Synthesis and Biological Evaluation of Novel Benzylidene Thiazolo Pyrimidin-3(5H)-One Derivatives

Starting compound 1 was synthesized according to reference. 1 Benzylidene thiazole pyrimidin-3(5H)-ones were synthesized reactions of 1 with bromoacetic acid and various aryl-aldehydes in the same vessel via one-step, unlike studies in the literature. Quantum chemical parameters and full geometry optimizations for all compounds were computed using DFT based on B3LYP. Cytotoxic action potential of synthesized compounds was evaluated using trypan blue dye exclusion and MTT assays in different cell lines including adenocarcinoma alveolar basal epithelial-like adherent A549 cells, the colon adenocarcinoma HT-29 cells, prostate adenocarcinoma DU-145 cells, and diploid ARPE-19 retinal pigment epithelial cells. Embryotoxicity and genotoxicity assessments were performed on pluripotent human embryonal carcinoma NT2 and human lymphocyte cells, respectively. Compound A1 exhibited good anticancer activity on A549 and DU-145 cell lines, and the compounds including A3, 4, 6, and 9 induced cytotoxicity on A549 cells. The compounds A1-10 also showed a good biosafety profile at relatively lower concentrations.

Equilibrium, kinetic and thermodynamic studies of the adsorption of trypan blue dye by Pseudomonas sp. strain MM02 inactivated biomass

Equilibrium, kinetic and thermodynamic studies of the adsorption of trypan blue dye by Pseudomonas sp. strain MM02 inactivated biomass

Combination of metabolomics and network pharmacology analysis to decipher the mechanisms of total flavonoids of Litchi seed against prostate cancer.

To explore the underlying mechanism of total flavonoids of Litchi seed (TFLS) in treating prostate cancer (PCa). Cell Counting Kit-8 (CCK-8), EdU incorporation assay, trypan blue dye assay and colony formation assay were employed to evaluate the effect of TFLS on PCa in vitro. The xenograft mouse model was established to explore the anti-tumour effect of TFLS in vivo. Alterations in the metabolic profiles of the PC3 cells and mouse serum were obtained by untargeted metabolomics. Combination with metabolomics analysis and network pharmacology strategies, the potential targets were predicted and further validated by RT-qPCR. TFLS attenuated PCa progression both in vitro and in vivo. Metabolomics results yielded from cells and serum indicated that the anti-cancer effect of TFLS was correlated with synergistic modulation of five common metabolic pathways including glycerophospholipid metabolism, arginine and proline metabolism, glycine, serine and threonine metabolism, tryptophan metabolism and steroid biosynthesis. Using in silico prediction and RT-qPCR analysis, we further revealed that TFLS exerted anti-PCa activities via regulating the expressions of nine genes, including MAOA, ACHE, ALDH2, AMD1, ARG1, PLA2G10, PLA2G1B, FDFT1 and SQLE. TFLS suppressed tumour proliferation in PCa, which may be associated with regulating lipid and amino acid metabolisms.

When it is all blue during phacoemulsification!

Trypan blue dye–assisted capsulorrhexis has been found to be very useful and effective during cataract surgery. Although this technique is safe and simplifies capsulorrhexis, especially in mature white cataracts, inadvertent staining of posterior capsule and vitreous has been reported in patients with history of trauma, ocular surgery, and pseudoexfoliation. Compromised zonular integrity has been observed to be the most probable underlying etiology for such a complication. Here, we report a case of vitreous staining without any clinical evidence of zonular compromise preoperatively.

Bioassay Techniques in Entomological Research

BIOASSAY (Bio - life, Assay - determination) is the measurement of the potency of any stimulus (physical, chemical, biological, and psychological) and the reactions that it produces in living matter. In Entomology, to know about the potency, Specificity of any compounds, standardization of doses, and identify modes of action bioassay is needed. In addition to the basic bioassay methods such as poison food, dry film techniques, etc., depending upon the researcher’s necessity, they are modified or newly developed. Diet covering and diet incorporation for lepidopterans and homopterans have been developed as advancements over the poison food assay method, by using insecticide-incorporated artificial diet. WHO cone, Wire ball test, Susceptibility test, CDC bottle assay and MCD assay developed for mosquito resistance monitoring and Larval Packet Technique (LPT) for cattle ticks, Sandwich method, TIBS, and Adult vial test are advancements over dry film technique. In Thrips Insecticide Bioassay System (TIBS), treated vials can be stored for up to 3 weeks at 4˚C for future efficacy assays at the field level. Adult Vial Tests have been developed for pentatomid and mirid adult bug bioassays. Larval Immersion Technique (LIT), Adult Immersion Technique (AIT) for cattle ticks, and MEBS have been developed as advancements over topical bioassay. In Mite Egg Bioassay System (MEBS), entomopathogenic fungi have been tested against Tetranychus cinnabarinus. MTT colorimetric assay and Trypan Blue Dye Exclusion assay helps to identify cell viability in insect cell line cultures. Spodoptera frugiperda cell line, Sf – 21 was tested against a scorpion, Leiurus quinquestriatus toxin β– BUTX – LQQ1a and the toxicity was determined through the viability of cell lines. Various basic and advanced bioassay techniques are discussed elaborately in this review.

P-142 Alterations in mitochondrial DNA levels in luteal granulosa cells may affect the morphology of mature oocytes and subsequently their fertilization potential: a retrospective multicenter study

Abstract Study question Do alterations in mitochondrial DNA levels in luteal granulosa cells (LGCs) affect the quality of mature oocytes in infertile women? Summary answer Alterations in the mitochondrial DNA levels in LGCs may modify the characteristics of the first polar body in oocytes and subsequently decrease their fertilization potential. What is known already Assisted reproductive treatment outcomes are closely linked to the oocyte quality as shown in several systematic reviews and meta-analyses. Particularly, the oocyte quality is highly affected by the surrounding luteal granulosa cells (LGCs). This is mainly due to the crucial role of the mitochondria in the LGCs in fueling the metabolic processes required for oocyte maturation. Therefore, modifications in the quality of LGCs may have direct effects on the developmental competence of oocytes. However, the exact mechanisms by which this could happen are not fully understood. Study design, size, duration A retrospective multicenter study was conducted on 303 mature oocytes retrieved from 51 women undergoing intracytoplasmic sperm injection (ICSI). It was conducted in Lebanon at Al Hadi IVF center and Azoury IVF clinic, between January 2019 and January 2020. G-Power 3.1 was used to determine the sample size for Generalized Linear Mixed Models through a one-sample t-test power analysis with alpha 0.05, power 0.8, medium effect size (f2 = 0.15), and 5 predictors. Resulting sample size: 43. Participants/materials, setting, methods This study excluded cases of premature ovarian failure, severe oligozoospermia (&amp;lt;2 x 102/ml), or cases using frozen gametes. Mature oocytes, from young women (&amp;lt; 36 years old), were injected and cultured in the Embryoscope where morphometric measurements were performed. LGCs vitality and mitochondrial DNA (mtDNA) levels were analyzed using trypan blue exclusion dye and next-generation sequencing, respectively. Possible associations between these independent variables and the size and integrity of the first polar body were evaluated. Main results and the role of chance The included women presented with primary infertility. Their mean age was 29.98 ± 5.5 years old and their mean body mass index was 23.33 ± 3.32 Kg/m2. 10% of the included women had polycystic ovary syndrome, and 48% were smokers. Regarding the LGCs parameters, a statistically significant negative correlation was found between the mtDNA levels in LGCs and their vitality percentage (r = −0.313, p = 0.029). Interestingly, the average size of the first polar body (PBI) was 346.67 µm. Here we found that the PBI size decreased by 15.05 units when the mtDNA level in LGCs increased by one unit above its average. In parallel, the median percentage of oocytes having a fragmented PBI was 33.33 (0-100). This percentage increased by 4.26% for every one unit increase in the mtDNA level in LGCs (p &amp;lt; 0.0001). In contrast, this percentage decreased by 0.86% for every unit increase in the percentage of LGCs vitality (p &amp;lt; 0.0001). Moreover, the mean percentage of fertilization rate was found to be 70.19 ± 26.7. A higher percentage of fragmented PBI (38.3%) was found among oocytes that did not fertilize compared to those that did successfully fertilize (24%) (p = 0.019). Limitations, reasons for caution This study only analyzed the effects of alterations in mtDNA levels in LGCs on oocytes. It would be worthwhile to also analyze the effects on foetal development and offspring health. Furthermore, it would be interesting to study the relationship between alterations in LGCs and oocyte quality within individual follicles. Wider implications of the findings Several studies have suggested that abnormal size and fragmentation of the PBI may impair embryo development. Therefore, treating factors that affect LGCs, such as obesity and polycystic ovary syndrome, may benefit infertile women. This highlights the need for new clinical trials in this context. Trial registration number Not applicable

The cytotoxicity effect of 7-hydroxy-3,4-dihydrocadalene from Heterotheca inuloides and semisynthetic cadalenes derivates towards breast cancer cells: involvement of oxidative stress-mediated apoptosis.

Heterotheca inuloides, traditionally employed in Mexico, has demonstrated anticancer activities. Although it has been proven that the cytotoxic effect is attributed to cadinane-type sesquiterpenes such as 7-hydroxy-3,4-dihydrocadalene, the mechanism of action by which these agents act in tumor lines and their regulation remain unknown. This study was undertaken to investigate for first time the cytotoxic activity and mechanism of action of 7-hydroxy-3,4-dihydrocadalene and two semi-synthetic cadinanes derivatives towards breast cancer cells. Cell viability and proliferation were assayed by thiazolyl blue tetrazolium bromide (MTT) assay and Trypan blue dye exclusion assay. Cell migration measure was tested by wound-healing assay. Moreover, the reactive oxygen species (ROS) and lipid peroxidation generation were measured by 2',7'-dichlorofluorescein diacetate (DCFH-DA) assay and thiobarbituric acid reactive substance (TBARS) assay, respectively. Furthermore, expression of caspase-3, Bcl-2 and GAPDH were analyzed by western blot. The results showed that 7-hydroxy-3,4-dihydrocadalene inhibited MCF7 cell viability in a concentration and time dependent manner. The cytotoxic potency of semisynthetic derivatives 7-(phenylcarbamate)-3,4-dihydrocadalene and 7-(phenylcarbamate)-cadalene was remarkably lower. Moreover, in silico studies showed that 7-hydroxy-3,4-dihydrocadalene, and not so the semi-synthetic derivatives, has optimal physical-chemical properties to lead a promising cytotoxic agent. Further examination on the action mechanism of 7-hydroxy-3,4-dihydrocadalene suggested that this natural product exerted cytotoxicity via oxidative stress as evidenced in a significantly increase of intracellular ROS levels and in an induction of lipid peroxidation. Furthermore, the compound increased caspase-3 and caspase-9 activities and slightly inhibited Bcl-2 levels. Interestingly, it also reduced mitochondrial ATP synthesis and induced mitochondrial uncoupling. Taken together, 7-hydroxy-3,4-dihydrocadalene is a promising cytotoxic compound against breast cancer via oxidative stress-induction.

Lactoferrin functionalized concave cube Au nanoparticles as biocompatible antibacterial agent

Gold nanoparticles (AuNPs) are one of the most extensively studied nanomaterials and their distinct physicochemical properties make them suitable for versatile applications. Herein, we synthesized concave cube-shaped gold nanoparticles (CCAuNPs) and functionalized them with lactoferrin (Lf), a natural antimicrobial protein, through electrostatic interaction as well as weak covalent formation. The functionalization of CCAuNPs was confirmed through UV–Visible (i.e., bathochromic shift of the surface plasmon resonance peak by 7 nm), Fourier transform infrared (FTIR) and X-ray diffraction (XRD) spectroscopy, and their surface zeta potential. The concave cusp of CCAuNPs was confirmed through atomic force microscopy (AFM). The Lf-functionalized CCAuNPs (Lf-CCAuNPs) exhibited superior antibacterial propensity against a series of bacteria when compared to that of CCAuNPs. However, they didn't demonstrate any antibacterial activity against Lactobacillus plantarum, one of the key beneficial gut bacteria. The lipid peroxidation (LPO) assay confirmed the oxidation of fatty acids in the bacterial membrane upon interaction with AuNPs, which made the bacterial membrane porous. The resultant membrane-impaired dead bacteria were visualized through CellTox™ Green assay as well as the Trypan Blue dye exclusion assay. Both the nanoparticles demonstrated excellent hemocompatibility as well as biocompatibility (both in vitro and in vivo) as confirmed by MTT assay and the levels of important functional biomarkers of liver (e.g., ALT, AST, and ALP) and kidney (e.g., creatinine, uric acid, and BUN) in the serum. Overall, Lf-CCAuNPs with excellent hemocompatibility, and biocompatibility can be deployed as potential antibacterial agents to tackle the menace of pathogenic bacteria.

IN VITRO ANTIOXIDANT AND ANTICANCER ACTIVITY OF THE FRUIT EXTRACT OF RIVINA HUMILIS L. (RED PIGEON BERRY)

The purpose of this study was to evaluate the cytotoxic and antioxidant properties of a hydromethanolic extract of Rivina humilis L fruits extract (HMERH) on human cancer cell lines and various in vitro models. HMERH concentrations ranging from 5 to 400mg/ml were utilized in several assay methods such as hydroxyl radicals, DPPH, superoxide radical scavenging activity, metal chelating, and nitric oxide radical test, ABTS, and reducing power assay. All antioxidant assays employed ascorbic acid as the standard. Its short-term cytotoxicity on EAC and DLA cells was determined using the trypan blue dye exclusion method, as well as the lactate dehydrogenase (LDH) leakage assay and in vitro cytotoxicity on MEF-L929, DU-145, and PC-3 cells using the MTT assay. The total antioxidant potential was also determined using the phosphomolybdenum test. HMERH demonstrated considerable cytotoxicity against all cancer cell lines examined. The findings of the free radical scavenging activities demonstrated a concentration-dependent and antiradical activity caused by the reduction of ABTS, DPPH, NO, OH, and SO radicals to non-radical form. The findings reveal that antioxidants and cytotoxic agents have potent antioxidant and cytotoxic effects on cancer cell lines. Hence, this fruit extract has the potential to be a natural antioxidant and anticancer drug source, which could be valuable in the development of new anticancer therapies.