Search Results (33)

Background: Many widely used drugs—including non-steroidal anti-inflammatory drugs, tetracyclines and thiazide diuretics—are labelled as photosensitising, yet their phototoxicity has been defined almost entirely by ultraviolet A (UVA) exposure. Whether this risk extends to the near-infrared (NIR) wavelengths of lasers commonly used in aesthetic medicine is unclear, and a history of such medication is often treated as a relative contraindication to laser procedures. Methods: A bibliometric analysis of 391 Scopus-indexed publications using VOSviewer was performed to quantitatively map the thematic structure of the retrieved literature. The bibliometric mapping was complemented by sequential refinement of the Scopus corpus and a focused assessment of the final records to determine whether studies directly addressing the predefined experimental question could be identified. We assessed whether 980 nm diode-laser irradiation of normal human dermal fibroblasts (HDFs) induces phototoxicity in the presence of seven photosensitisers (ketoprofen, meloxicam, hydrochlorothiazide, doxycycline and diclofenac, with chlorpromazine and 8-methoxypsoralen as reference photosensitizers). Cells were irradiated through the closed culture-plate lid at two fluences (62.3 and 34.4 mJ/cm and viability was measured 0.5, 1 and 24 h later. Data were analysed by two-way ANOVA (compound × laser), pairwise Welch tests with Benjamini–Hochberg correction, and equivalence testing against an ISO 10993-5-based margin. Results: Viability remained between 82.7% and 109.6% of control under all conditions and never fell below the 70% cytotoxicity threshold. The compound × laser interaction was non-significant at every time point (p = 0.25–0.69), indicating no drug-specific phototoxicity, and no pairwise comparison remained significant after correction. A small, drug- and fluence-independent reduction of about six percentage points was observed but stayed well within the non-cytotoxic range and was equally present in drug-free controls. Conclusions: Under the specific 980 nm irradiation conditions tested, the photosensitising drugs did not increase phototoxicity in normal human dermal fibroblasts. These findings support a wavelength-specific view of drug photosafety: rigorous UVA-directed photoprotection remains essential, whereas the tested drugs did not increase phototoxicity under the specific in vitro 980 nm irradiation conditions evaluated in this study. Full article

Background/Objectives: Extracellular vesicles (EVs) have emerged as important mediators of intercellular communication in cardiovascular disease, particularly through the transfer of regulatory microRNAs. However, direct evidence of cytosolic microRNA delivery by mesenchymal stem cell (MSC)-derived EVs into cardiomyocytes remains limited. Many prior studies have inferred transfer without fully excluding endogenous microRNA induction or surface-bound artifacts. The objective of this study was to determine whether MSC-derived EV microRNA undergoes functional cytoplasmic delivery into cardiomyocytes and to identify the endocytic pathways involved. Methods: To eliminate background from endogenous microRNA expression, MSC-derived EVs were loaded with the exogenous microRNA cel-miR-39-3p. H9C2 cardiomyocytes were treated with increasing vesicle doses, and intracellular microRNA levels were quantified using real-time polymerase chain reactions. Cytosolic accessibility was assessed indirectly using a cholesterol-modified antagomir targeting cel-miR-39-3p. Pharmacologic inhibitors of macropinocytosis (5-(N-ethyl-N-isopropyl) amiloride (EIPA)) and clathrin-mediated endocytosis (chlorpromazine and ES9-17) were applied to delineate uptake pathways, with transferrin assays confirming clathrin inhibition. Results: EV treatment produced a dose-dependent increase in intracellular cel-miR-39-3p levels. Antagomir administration reduced intracellular microRNA levels by 89%, providing indirect functional evidence that transferred cel-miR-39-3p reached an antagomir-accessible cytosolic compartment. At the highest preparation dose, EIPA and ES9-17 reduced cell-associated cel-miR-39-3p by 71% and 36%, respectively, consistent with contributions from EIPA-sensitive and clathrin-associated uptake processes. Notably, chlorpromazine increased microRNA accumulation despite blocking transferrin uptake, suggesting compensatory or membrane curvature-dependent effects. Conclusions: These findings provide strong functional evidence that exogenous cel-miR-39-3p associated with MSC EV preparations reaches an antagomir-accessible cytoplasmic compartment in H9C2 cardiomyocytes and implicate EIPA-sensitive uptake consistent with macropinocytosis and clathrin-associated uptake. Additional genetic, localization, and pulse-chase studies are required to establish the precise intracellular trafficking mechanisms. This work strengthens the mechanistic foundation for EV-based therapies and informs the rational development of cell-free strategies for cardiovascular disease. Full article

A naive Bayesian classifier (NBC) combined with principal component analysis (PCA) effectively differentiates the dose-dependent effects of certain groups of psychoactive drugs based on their impact on the amplitude–spectral characteristics of electrocorticograms (ECoG) in rats. This approach has been shown to be useful for pharmacological screening of agents with unknown or poorly understood activity. Despite previously obtained optimistic results, classification determination for some drugs was inaccurate, necessitating the search for possible ways to improve the predictive effectiveness of the proposed algorithm. One possible approach would be to use as input quantitative data not only the impact of the psychoactive drugs studied on the amplitude–spectral characteristics of ECoG but also connectivity changes, including the average coherence power of different pairs of leads. The aim of this study was to compare the accuracy of NBC in classifying the pharmacological mechanism of action of agents with well-known mechanisms (test set) using pharmaco-EEG data on changes in the amplitude–spectral characteristics of ECoG, coherence, and the combined use of two data sets. Materials and methods. Experiments were performed on Wistar rats with chronically implanted ECoG electrodes. The training set, relative to which the effects of the pharmacological agents from the test set were classified, were the matrices of effects of 12 pharmacological agents: the NMDA antagonist dizocilpine, the D2/D3 antagonists haloperidol and sulpiride, the M-anticholinergic tropicamide, the H1/5HT2A receptor blocker hydroxyzine, the acetylcholinesterase inhibitor galantamine, the alpha-2 adrenergic agonist dexmedetomidine, the alpha-2 adrenergic antagonist atipamezole, the adenosine receptor blocker caffeine and the GABA-mimetics aminophenylbutyric acid (phenibut), bromdihydrochlorophenylbenzodiazepine (phenazepam) and 5-ethyl-5-phenyl-2,4,6(1H,3H,5H)-pyrimidinetrione. The test set included various drugs with tropism for the targets of the training set drugs: dopamine receptor antagonists chlorpromazine, droperidol, tiapride and raclopride, H1-histamine blockers diphenhydramine and promethazine, 5-HT2-receptor blockers ritanserin and glemenserin, acetylcholinesterase inhibitor ipidacrine, alpha2-adrenergic receptor antagonist yohimbine, alpha2-adrenergic agonists medetomidine and xylazine, GABA-mimetics 5-ethyl-5-(1-methylbutyl)-2,4,6(1H,3H,5H)-pyrimidinetrione and chloral hydrate. The analysis of the ECoG signal included the calculation of 132 amplitude–spectral characteristics and 75 coherence indicators, which, using the PCA, led to new integrative indicators used for further classification of the NBC. Results and discussion. For each drug in the test set, the median similarity probability with a particular group from the training set was calculated, which was used to assess the classification quality. It was found that, when using the amplitude–spectral characteristics of ECoG, the proposed methodological approach allows for the identification of the ECoG effects of several groups of psychoactive drugs, including D2/D3-dopamine, M-cholinergic, H1-histamine, and 5-HT2-serotonin receptor blockers, AChE inhibitors, GABA-mimetics, and alpha-2-adrenergic receptor agonists and antagonists. This approach enabled the correct classification of 18 of 24 groups in the test set. When using changes in coherence indices as the initial data, the classification accuracy also amounted to 18 of 24 groups. When combining the two data sets, the number of correctly identified NBC groups was 20 of 24 groups. When comparing the classification during training (confusion matrix), it was found that coherence data or adding coherence data to the data based on changes in amplitude–spectral characteristics leads to a statistically significant (p < 0.01 in both cases) increase in accuracy. Conclusions. The obtained data demonstrated high accuracy in classifying the pharmacological activity of the test sample drugs using any of the three compared approaches. Despite the lack of statistically significant differences between them, classification based on the combined dataset demonstrated a higher number of “correct” similarities. This allows us to recommend the approach based on combined data of drug effects on amplitude–spectral characteristics and coherence as the most promising for further studies using pharmaco-EEG screening. Full article

Background/Objectives: Hydrazones are organic compounds with the general structure R₂C=NNHR₁, distinguished by their versatility and modifiability, and are widely used in various applications due to their physicochemical and biological properties. They exhibit anticancer, anti-inflammatory, antibiofilm, and antibacterial activities. Antibiotic-resistant bacteria pose a serious public health threat, employing mechanisms such as enzymatic inactivation and efflux pumps. This study evaluated the antibacterial activity of the hydrazone HDZH1,4BENZ, a hydralazine-derived compound, as well as its potential adjuvant effect in combination with antibiotics against Staphylococcus aureus strains expressing efflux pumps. Methods: The strains used were 1199B (NorA efflux pump-expressing) and K2068 (MepA efflux pump-expressing). All assays were conducted using the broth microdilution method in Brain Heart Infusion (BHI) medium. Initially, the intrinsic antibacterial activity of the compound was determined. Subsequently, modulation assays were performed to evaluate its potential effect on efflux pump activity, with a standard efflux pump inhibitor included as a positive control. Results: Although HDZH1,4BENZ did not demonstrate significant direct antibacterial activity, the results indicate that this hydrazone exerts a notable inhibitory effect on the NorA (Norfloxacin resistance efflux pump A) and MepA (Multidrug efflux protein A) efflux pumps in S. aureus, thereby enhancing the efficacy of antibacterial agents. Conclusions: The activity of the hydrazone was comparable to that of chlorpromazine, suggesting that it may represent a promising alternative in the fight against antibiotic-resistant bacterial infections. Full article

Background/Objectives: Epilepsy is a major neurological disorder associated with significant comorbidity and treatment challenges. In low- and middle-income countries, access to newer antiseizure medications (ASMs) remains limited, and prescription patterns often rely on older agents. This study aimed to characterize national prescribing patterns of ASMs among patients with epilepsy in Kazakhstan from 2021 to 2023. Methods: We conducted a retrospective observational study using de-identified electronic health record data from the Unified National Electronic Health System of Kazakhstan. All patients with an ICD-10 diagnosis of epilepsy (G40) and at least one ASM prescription during 2021–2023 were included. Prescription frequencies, therapy type, and chronic polytherapy levels were analyzed. Associations between therapy type, age, and comorbidity status were determined. Results: A total of 54,274 patients were identified (median age 42 years; interquartile range (IQR) 31–57). Monotherapy predominated: 61.7% remained on monotherapy, 18.5% remained on polytherapy, and 19.8% had mixed exposure. Carbamazepine and valproic acid were most frequently prescribed (64.3% and 45.6% of patients, respectively). Among those with chronic medication data (n = 15,752), nervous-system drugs were common (70.1%), led by psycholeptics (49.7%); frequently dispensed agents included chlorpromazine (n = 5991), clozapine (n = 1875), and risperidone (n = 1642). Cardiovascular agents were recorded in 37.2% (acetylsalicylic acid n = 4056; atorvastatin n = 2235), and diabetes drugs in 12.1% (metformin n = 1430). Conclusions: Epilepsy treatment in Kazakhstan remains dominated by older broad-spectrum ASMs, while the use of lamotrigine and levetiracetam is steadily increasing. The frequent co-prescription of psychotropic and cardiometabolic drugs underscores the need for coordinated, multidisciplinary care and continued monitoring of prescribing patterns to enhance treatment safety and effectiveness. Full article

Although chlorpromazine is primarily used in psychiatry, it has been shown since its introduction to influence the course of neoplastic diseases. According to the strategy of drug repurposing, chlorpromazine has been successfully tested for its potential antitumor effects on multiple cancer cell lines. This effect is consistent with the overlap of molecular pathways observed for years between schizophrenia and cancer. The main objective of this work was to evaluate the lipophilicity of 17 previously synthesized tetracyclic chlorpromazine analogues exhibiting diverse anticancer and antimicrobial activity using thin-layer chromatography and computational methods. For a compound to become an effective drug, it must have a favorable ADMET profile, which determines its pharmacokinetic properties as a drug candidate. Lipophilicity is one of the key parameters widely employed in designing new bioactive compounds as potential therapeutic agents. In this article, chromatographic plates precoated with silica gel 60 RP-18F₂₅₄ and a mixture of acetone and TRIS buffer were used as the mobile phase. The chromatographic parameter of lipophilicity (R_M0) of the investigated compounds determined by means of the Soczewinski–Wachtmeister formula was useful to obtain the values of the experimental lipophilicity parameter expressed as logP_TLC. The results of logP_TLC were compared with theoretical values of logP obtained using different algorithms (iLOGP, XLOGP3, WLOGP, MLOGP, SILCOS-IT, and ClogP). Furthermore, the online platforms, such as SwissADME and pkCSM, allowed the determination of the remaining ADME parameters of the quinoline derivatives of chlorpromazine. The study of lipophilicity and ADME factors enabled confirmation that the tested compounds demonstrated favorable properties. Therefore, they can be considered as promising starting structures for further studies. Full article

Background/Objectives: In patients with schizophrenia, gait disturbances (e.g., reduced walking speed and stride length) are linked to neural dysfunction and extrapyramidal symptoms. To inform gait rehabilitation strategies, this study examines the relationships of walking speed with extrapyramidal symptoms, stride length, antipsychotic dosage, ankle joint range of motion, and body composition in patients with chronic schizophrenia. Methods: Sixty-eight patients with chronic schizophrenia were included. All variables were described based on their measurement levels using non-parametric methods. Spearman’s rho was calculated to assess correlations. For multiple linear regression analyses, backward stepwise elimination was used to determine variables associated with walking speed. Statistical significance was set to p < 0.05. Results: Walking speed was positively correlated with stride length, chlorpromazine-equivalent dose, ankle plantar flexion, body mass index, bone mineral content, trunk muscle mass, and skeletal muscle mass index. In contrast, it was negatively correlated with drug-induced extrapyramidal symptoms scale (DIEPSS) scores for gait, bradykinesia, tremor, overall severity, and age. The multiple linear regression indicated that DIEPSS 2 bradykinesia level and ankle plantar flexion angle, adjusted for a 26% variance, best explained the walking speed. Conclusions: A lower bradykinesia severity and a higher ankle plantar flexion are associated with higher walking speeds. Thus, it is critical to assess stride length, bradykinesia, angle/limitation/torque of ankle plantar flexion, trunk and upper and lower limb muscle masses, and walking speed in patients with chronic schizophrenia. Specific strategies for gait rehabilitation should focus on stride training, plantar flexion strengthening exercises, and balance training. Full article

Background/Objectives: This study aimed to determine the association between chronic schizophrenia, extrapyramidal symptoms (EPSs), body composition, nutritional status, and dynapenia/sarcopenia. Methods: Data from 68 chronic patients with schizophrenia were analyzed using Spearman’s rho correlation coefficients, Kruskal–Wallis test, Mann–Whitney U test, and Cramér’s V statistics. Results: Among the participants, 32.4% had no loss of muscle mass or function, 39.7% had dynapenia, and 27.9% had sarcopenia. This study identified five key findings: (1) Bilateral grip strength, skeletal muscle index, and walking speed are interrelated, with higher negative symptom scores linked to slower movement and rigidity, particularly in the sarcopenia group, indicating that negative symptoms may contribute to muscle weakness and progression to sarcopenia. (2) Increasing age is associated with a decrease in chlorpromazine equivalent dose and an increase in the severity of EPSs. (3) Blood urea nitrogen (BUN)/creatinine ratio and all sarcopenia risk indicators were significantly negatively correlated. (4) Dynapenia and sarcopenia groups exhibited significant differences in muscle mass and nutritional status compared to the non-penia group, including reduced muscle mass, lower basal metabolic rate, and lower visceral fat levels. (5) There was an association between the Barthel Index (BI) score for activities of daily living (ADL) and dynapenia/sarcopenia. Particularly with regard to ADL, it seems necessary to pay attention to muscle weakness in partially independent patients who score 60 points or more. Conclusions: BUN/creatinine ratio, BI, EPSs, body mass index, grip strength, total protein, and albumin were useful indicators for detecting the risk of dynapenia/sarcopenia in routine psychiatric care. Full article

The utilization of membrane sensors for the monitoring and determination of pharmaceutical environmental pollutants has emerged as a crucial objective in recent years. Given the extensive use of chlorpromazine hydrochloride (CPZ) in medicine, its presence in the environment, particularly in surface water such as rivers, is highly probable. Prolonged exposure of river water to sunlight and the photo-degradability of CPZ may enhance its photo-degradation. For the purpose of measuring CPZ in the presence of its primary photo-degradants, two sensitive and selective membrane electrodes were developed. These were synthesized utilizing two ion-pairing agents: sodium tetraphenylborate (TPB) and phosphotungstic acid (PTA). The electrodes exhibited a linear range that extended from 1 × 10⁻⁶ M to 1 × 10⁻² M. The membrane electrodes of CPZ-TPB and CPZ-PTA exhibited slopes of 59.90 ± 0.60 mV/decade and 58.90 ± 0.80 mV/decade, respectively. The sensors mentioned above showed acceptable performance in a pH range of 2.0 to 6.0. All test parameters were optimized to provide superior electrochemical performance. The fabricated membranes were effectively employed to sensitively quantify CPZ in the presence of its principal photodegradants. The developed sensors were successfully employed to quantify CPZ in river water samples without necessitating pre-treatment procedures. Full article

Phenothiazine derivatives are widely studied in various fields such as biology, chemistry, and medicine research because of their pharmaceutical effects. The first compound used successfully in the treatment of psychosis was a phenthiazine derivative, chlorpromazine. Apart from its activity in neurons, chlorpromazine has also been reported to display anticancer and antibacterial properties. In this study, we present the synthesis and research on the activity of A549, MDA, MiaPaCa, PC3, and HCT116 cancer cell lines and of S. aureus, S. epidermidis, E. coli, and P. aeruginosa bacterial strains against a series of new tetracyclic chlorpromazine analogues containing a quinoline scaffold in their structure instead of the benzene ring and various substituents at the thiazine nitrogen. The structure of these novel molecules has been determined by ¹H NMR, ¹³C NMR, and HRMS spectral techniques. The seven most active of the twenty-four new chlorpromazine analogues tested were selected to study the mechanism of cytotoxic action. Their ability to induce apoptosis or necrosis in cancer cells was assessed by flow cytometry analysis. The results obtained confirmed the proapoptotic activity of selected compounds, especially in terms of inducing late apoptosis or necrosis in cancer cell lines A549, MiaPaCa-2, and HCT-116. Furthermore, studies on the induction of cell cycle arrest suggest that the new chlorpromazine analogues exert antiproliferative effects by inducing cell cycle arrest in the S phase and, consequently, apoptosis. Full article

Human adenovirus (HAdV) is a common pathogen, which can lead to various clinical symptoms and—in some cases—central nervous system (CNS) dysfunctions, such as encephalitis and meningitis. Although the initial events of virus entry have already been identified in various cell types, the mechanism of neuronal uptake of adenoviruses is relatively little understood. The aim of this study was to investigate early events during adenoviral infection, in particular to determine the connection between cellular coxsackievirus and adenovirus receptor (CAR), clathrin, caveolin, and early endosomal proteins (EEA1 and Rab5) with the entry of HAdVs into primary murine neurons in vitro. An immunofluorescence assay and confocal microscopy analysis were carried out to determine HAdV4, 5, and 7 correlation with CAR, clathrin, caveolin, and early endosomal proteins in neurons. The quantification of Pearson’s coefficient between CAR and HAdVs indicated that the HAdV4 and HAdV5 types correlated with CAR and that the correlation was more substantial for HAdV5. Inhibition of clathrin-mediated endocytosis using chlorpromazine limited the infection with HAdV, whereas inhibition of caveolin-mediated endocytosis did not affect virus entry. Thus, the entry of tested HAdV types into neurons was most likely associated with clathrin but not caveolin. It was also demonstrated that HAdVs correlate with the Rab proteins (EEA1, Rab5) present in early vesicles, and the observed differences in the manner of correlation depended on the serotype of the virus. With our research, we strove to expand knowledge regarding the mechanism of HAdV entry into neurons, which may be beneficial for developing potential therapeutics in the future. Full article

Here, we present evidence that caveolae-mediated endocytosis using LDLR is the pathway for SARS-CoV-2 virus internalization in the ocular cell line ARPE-19. Firstly, we found that, while Angiotensin-converting enzyme 2 (ACE2) is expressed in these cells, blocking ACE2 by antibody treatment did not prevent infection by SARS-CoV-2 spike pseudovirions, nor did antibody blockade of extracellular vimentin and other cholesterol-rich lipid raft proteins. Next, we implicated the role of cholesterol homeostasis in infection by showing that incubating cells with different cyclodextrins and oxysterol 25-hydroxycholesterol (25-HC) inhibits pseudovirion infection of ARPE-19. However, the effect of 25-HC is likely not via cholesterol biosynthesis, as incubation with lovastatin did not appreciably affect infection. Additionally, is it not likely to be an agonistic effect of 25-HC on LXR receptors, as the LXR agonist GW3965 had no significant effect on infection of ARPE-19 cells at up to 5 μM GW3965. We probed the role of endocytic pathways but determined that clathrin-dependent and flotillin-dependent rafts were not involved. Furthermore, 20 µM chlorpromazine, an inhibitor of clathrin-mediated endocytosis (CME), also had little effect. In contrast, anti-dynamin I/II antibodies blocked the entry of SARS-CoV-2 spike pseudovirions, as did dynasore, a noncompetitive inhibitor of dynamin GTPase activity. Additionally, anti-caveolin-1 antibodies significantly blocked spike pseudotyped lentiviral infection of ARPE-19. However, nystatin, a classic inhibitor of caveolae-dependent endocytosis, did not affect infection while indomethacin inhibited only at 10 µM at the 48 h time point. Finally, we found that anti-LDLR antibodies block pseudovirion infection to a similar degree as anti-caveolin-1 and anti-dynamin I/II antibodies, while transfection with LDLR-specific siRNA led to a decrease in spike pseudotyped lentiviral infection, compared to scrambled control siRNAs. Thus, we conclude that SARS-CoV-2 spike pseudovirion infection in ARPE-19 cells is a dynamin-dependent process that is primarily mediated by LDLR. Full article

Nasal drug delivery has been a focus of scientific interest for decades. A number of drug delivery systems and devices are available and have been highly successful in providing better and more comfortable therapy. The benefits of nasal drug delivery are not in question. The nasal surface provides an excellent context for the targeted delivery of active substances. In addition to the large nasal surface area and intensive absorption, the active substances delivered through the nose overcome the blood–brain barrier and can be delivered directly to the central nervous system. Formulations for nasal administration are typically solutions or liquid dispersed systems such as emulsions or suspensions. Formulation techniques for nanostructures have recently undergone intensive development. Solid-phase heterogeneous dispersed systems represent a new direction in pharmaceutical formulations. The wide range of possible examples and the variety of excipients allow for the delivery of a wide range of active ingredients. The aim of our experimental work was to develop a solid drug delivery system that possesses all of the above-mentioned advantageous properties. In developing solid nanosystems, we not only exploited the advantages of size but also the adhesive and penetration-enhancing properties of excipients. During formulation, several amphiphilic compounds with adhesion properties and penetration enhancing effects were incorporated. We used chlorpromazine (CPZ), which is mainly used in the treatment of psychotic disorders such as schizophrenia and bipolar disorder. Chlorpromazine has been previously investigated by our team in other projects. With the availability of previous methods, the analytical characterization of the drug was carried out effectively. Due to the frequent and severe side effects of the drug, the need for therapeutic dose reduction is indisputable. In this series of experiments, we succeeded in constructing drug delivery systems. Finely divided Na nanoparticles were formed using a Büchi B90 nanospray dryer. An important step in the development of the drug carrier was the selection of suitable inert carrier compounds. Particle size determination and particle size distribution analysis were performed to characterize the prepared nanostructures. As safety is the most important aspect of any drug formulation, all components and systems were tested with different biocompatibility assays. The tests performed demonstrated the safe applicability of our systems. The bioavailability of chlorpromazine was studied as a function of the ratio of the active ingredient administered nasally and intravenously. As described above, most nasal formulations are liquids, but our system is solid, so there is currently no tool available to accurately target this system. As a supplement of the project, a nasal dosing device was developed, corresponding to the anatomical structure; a prototype of the device was made using 3D FDM technology. Our results lay the foundation for the design and industrial scaling of a new approach to the design and production of a high-bioavailability nasal medicinal product. Full article

In this work, we studied the induction of somatic embryogenesis in Arabidopsis using IZEs as explants. We characterized the process at the light and scanning electron microscope level and studied several specific aspects such as WUS expression, callose deposition, and principally Ca²⁺ dynamics during the first stages of the process of embryogenesis induction, by confocal FRET analysis with an Arabidopsis line expressing a cameleon calcium sensor. We also performed a pharmacological study with a series of chemicals know to alter calcium homeostasis (CaCl₂, inositol 1,4,5-trisphosphate, ionophore A23187, EGTA), the calcium–calmodulin interaction (chlorpromazine, W-7), and callose deposition (2-deoxy-D-glucose). We showed that, after determination of the cotiledonary protrusions as embryogenic regions, a finger-like appendix may emerge from the shoot apical region and somatic embryos are produced from the WUS-expressing cells of the appendix tip. Ca²⁺ levels increase and callose is deposited in the cells of the regions where somatic embryos will be formed, thereby constituting early markers of the embryogenic regions. We also found that Ca²⁺ homeostasis in this system is strictly maintained and cannot be altered to modulate embryo production, as shown for other systems. Together, these results contribute to a better knowledge and understanding of the process of induction of somatic embryos in this system. Full article

Rapid and highly sensitive quantitative analysis of chlorpromazine (CPZ) in human whole blood is of great importance for human health. Herein, we utilize the screen-printed carbon electrodes (SPCE) as the electrode substrates for growth of highly electroactive and antifouling nanocomposite materials consisting of vertically ordered mesoporous silica films (VMSF) and electrochemically reduced graphene oxide (ErGO) nanosheets. The preparation of such VMSF/ErGO/SPCE could be performed by using an electrochemical method in a few seconds and the operation is controllable. Inner ErGO layer converted from graphene oxide (GO) in the growth process of VMSF provides oxygen-containing groups and two-dimensional π-conjugated planar structure for stable fabrication of outer VMSF layer. Owing to the π-π enrichment and excellent electrocatalytic abilities of ErGO, electrostatic preconcentration and antifouling capacities of VMSF, and inherent disposable and miniaturized properties of SPCE, the proposed VMSF/ErGO/SPCE sensor could be applied for quantitative determination of CPZ in human whole blood with high accuracy and sensitivity, good stability, and low sample consumption. Full article