Peptide-drug conjugate designated for targeted delivery to HER2-expressing cancer cells.

Targeted therapy of the highest globally incident breast cancer shall resolve the issue of off-target toxicity concurring with augmented killing of specific diseased cells. Thus, the goal of this study was to prepare a peptide-drug conjugate targeting elevated expression of HER2 receptors in breast cancer. Towards this, the rL-A9 peptide was conjugated with the chemotherapeutic drug doxorubicin (DOX) through a N-succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) linker. The synthesized peptide-drug conjugate, rL-A9-DOX, was characterized by mass spectrometry. Molecular docking studies, based on binding energy data, suggested a stronger interaction of rL-A9-DOX with the HER2 receptor in comparison to the unconjugated peptide, rL-A9. The cytotoxic effect of the rL-A9-DOX conjugate was observed to be higher in HER2-positive SKOV3 cells compared to HER2-negative MDA-MB-231 cells, indicating selective cell killing. Cellular internalization of the rL-A9-DOX conjugate was evident from the flow cytometry analysis, where a noticeable shift in mean fluorescent intensity (MFI) was observed for the conjugate compared to the control group. This data was further validated by confocal microscopy, where the fluorescent signal ascertained nuclear accumulation of rL-A9-DOX. The present studies highlight the promising potential of rL-A9-DOX for targeted delivery of the drug into a defined group of cancer cells.

Poison in the water: Arsenic's silent assault on fish health.

Arsenic occurs across the world in freshwater and marine environments, menacing the survival of aquatic organisms. Organic and inorganic forms of this substance can be found, in which the inorganic form is more hazardous than the organic form. Most aquatic bodies contain inorganic arsenic species, but organic species are believed to be the dominant form of arsenic in the majority of fish. Natural and anthropogenic both are the sources of water contamination with arsenic. Its bioaccumulation and transfer from one trophic level to another in the aquatic food chain make arsenic a vital environmental issue. Continuous exposure to low concentrations of arsenic in aquatic organisms including fish leads to its bioaccumulation, which may affect organisms of higher trophic levels including large fishes or humans. Humans can be exposed to arsenic through the consumption of fish contaminated with arsenic. Hence, the present review facilitates our understanding about sources of arsenic, its bioaccumulation, food chain transfer, and its effect on the fish health. Also, "Poison in the Water: Arsenic's Silent Assault on Fish Health" serves as a wake-up call to recognize the pressing need to address arsenic contamination in water bodies. By understanding its devastating impact on fish health, we can strive to implement sustainable practices and policies that safeguard our precious aquatic environments and ensure the well-being of both wildlife and human communities that depend on them.

Loss of CpFTSY Reduces Photosynthetic Performance and Affects Insertion of PsaC of PSI in Diatoms.

The chloroplast signal recognition particle (CpSRP) receptor (CpFTSY) is a component of the CpSRP pathway that post-translationally targets light-harvesting complex proteins (LHCPs) to the thylakoid membranes in plants and green algae containing chloroplasts derived from primary endosymbiosis. In plants, CpFTSY also plays a major role in the co-translational incorporation of chloroplast-encoded subunits of photosynthetic complexes into the thylakoids. This role has not been demonstrated in green algae. So far, its function in organisms with chloroplasts derived from secondary endosymbiotic events has not been elucidated. Here, we report the generation and characterization of mutants lacking CpFTSY in the diatom Phaeodactylum tricornutum. We found that this protein is not involved in inserting LHCPs into thylakoid membranes, indicating that the post-translational part of the CpSRP pathway is not active in this group of microalgae. The lack of CpFTSY caused an increased level of photoprotection, low electron transport rates, inefficient repair of photosystem II (PSII), reduced growth, a strong decline in the PSI subunit PsaC and upregulation of proteins that might compensate for a non-functional co-translational CpSRP pathway during light stress conditions. The phenotype was highly similar to the one described for diatoms lacking another component of the co-translational CpSRP pathway, the CpSRP54 protein. However, in contrast to cpsrp54 mutants, only one thylakoid membrane protein, PetD of the Cytb6f complex, was downregulated in cpftsy. Our results point to a minor role for CpFTSY in the co-translational CpSRP pathway, suggesting that other mechanisms may partially compensate for the effect of a disrupted CpSRP pathway.

Synthesis and 177Lu Labeling of the First Retro Analog of the HER2-Targeting A9 Peptide: A Superior Variant.

The retro analog of the HER2-targeting A9 peptide was synthesized by coupling amino acids in a reverse fashion and switching the N-terminal in the original sequence of the L-A9 peptide (QDVNTAVAW) to the C-terminal in rL-A9 (WAVATNVDQ). Modification in the backbone resulted in higher conformational stability of the retro peptide as evident from CD spectra. Molecular docking analysis revealed a higher HER2 binding affinity of [177Lu]Lu-DOTA-rL-A9 than the original radiopeptide [177Lu]Lu-DOTA-L-A9. Enormously enhanced metabolic stability of the retro analog led to significant elevation in tumor uptake and retention. SPECT imaging studies corroborated biodistribution results demonstrating a remarkably higher tumor signal for [177Lu]Lu-DOTA-rL-A9. The presently studied retro probe has promising efficiency for clinical screening.

Regulation of omega-3 fatty acids production by different genes in freshwater fish species: a review.

The present study aims to compare the gene expression of three different fish species (common carp, tilapia, and trout) with varying levels of fatty acids (FA). Based on transcriptome analysis and RNA sequencing, various genes and their associated metabolic pathways are identified. Pathways are categorized based on the genes they encode. Genes that were differentially expressed and their promoter's methylation patterns were revealed by RNA-seq analysis in common carp. Furthermore, fatty acid-enriched pathways, such as ARA4 and adipocytokine signaling, were also identified. Many genes and pathways may influence tilapia's growth and omega-3 content. Using the mTOR pathway, trout with differential expression were discovered to be involved in producing omega-3 fatty acids. This study revealed major pathways in fish species to produce omega-3 fatty acids.

Adsorptive removal of oxytetracycline using MnO2-engineered pine-cone biochar: thermodynamic and kinetic investigation and process optimization.

Indiscriminate use of oxytetracycline is linked to the development of antibiotic-resistant genes, posing a serious threat to human health and ecosystem balance. This article reports the adsorptive elimination of oxytetracycline (OTC) from aqueous solution using a newly developed MnO2-modified pine-cone biochar (MnO2/PCBC). The MnO2/PCBC was characterized using X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy, CHNS analyzer, inductively coupled plasma-optical emission spectroscopy, and Brunauer-Emmett-Teller N2 adsorption analyzer. Batch adsorption experiments, designed using the central composite design framework of response surface methodology, were conducted to investigate the influence of process variables on the adsorption of OTC onto MnO2/PCBC. The optimized conditions for achieving maximum removal (88.1%) were found to be at pH 8, MnO2/PCBC dose 0.44 g/L, initial OTC concentration 200 mg/L, and temperature 303 K. The adsorption process follows Langmuir (R2=0.95) and Freundlich (R2=0.95) isotherms and pseudo-second-order (R2=0.99) adsorption kinetics. The adsorption process was found to be endothermic (ΔH0 = 33.04 kJ/mol) and spontaneous in nature (ΔG0 from -1.33 kJ/mol at 283 K to -5.65 kJ/mol at 313 K). The synthesized MnO2/PCBC could be recycled and reused for OTC removal with a percentage removal of around 80% after fifth cycle. The results indicate an effective removal of oxytetracycline with only 0.44 g/L MnO2/PCBC with maximum adsorption capacity of 357.14 mg/g which demonstrates improved performance in comparison to many adsorbents reported in literature. This implies that MnO2/PCBC offers potential to be developed into a cost-effective technique for antibiotic removal from water.

Assessment of 177 Lu-labeled carboxyl-terminated polyamidoamine (PAMAM) dendrimer-RGD peptide conjugate.

Structurally unique polyamidoamine (PAMAM) dendrimers implanted with targeting biological moiety along with complexed radiometal constitute a favorable nano-system for diagnosis and therapy of targeted tumor sites. In the present study, carboxyl functionalities of PAMAM- generation 4 dendrimer (PAMAM-G4-COOH) were conjugated with ε-amino group of lysine of cRGDfK peptide to impart integrin αv ß3 targeting capability. Reaction of p-NH2 -Bn-DOTA with PAMAM was accomplished via acid-amine coupling using EDC/NHS for 177 Lu-complexation. 177 Lu-labeled nano-system, 177 Lu-PAMAM-DOTA-cRGDfK demonstrated receptor-mediated uptake in murine melanoma B16F10 cells during in vitro cell uptake studies. In vivo biodistribution studies demonstrated low tumor uptake and retention of 177 Lu-PAMAM-DOTA-cRGDfK which may be attributed to rapid blood clearance. However, fast clearance from non-target organs resulted in higher target to background ratio. Tumor uptake of targeted nano-system, 177 Lu-PAMAM-DOTA-cRGDfK was observed to be significantly (p < 0.05) higher in comparison to 177 Lu-PAMAM-DOTA without the targeting peptide. Inhibition studies with unlabeled cRGDfK resulted in 60% reduction in tumor uptake of 177 Lu-PAMAM-DOTA-cRGDfK, indicating specificity of the developed nano-system towards integrin αv ß3 receptors.

Current synthesis and characterization techniques for clay-based polymer nano-composites and its biomedical applications: A review.

Clays and its composites have received considerable attention recently due to their low cost, wide availability and low environmental impact. The development of various preparation processes and applications of innovative polymer-nanoclay composites has been aided by recent breakthroughs in material technologies. Novel polymer-nanoclay composites with better qualities have been effectively adopted in a variety of fields, including aerospace, car, construction, petroleum, biomedical, and wastewater treatment, owing to innovative production processes. Due to their superior qualities, such as increased density, strength, relatively large surface areas, high elastic modulus, flame retardancy, and thermomechanical/optoelectronic/magnetic capabilities, these composites are acknowledged as potential advanced materials. Hence the present paper reviews the advances in synthesis and preparation of clay-polymer nanocomposites. In addition, this study also focuses on the various techniques used for clay-polymer nanocomposites characterization e.g. scanning electron microscope (SEM), transmission electron microscope (TEM), thermo-gravimetric analysis (TGA) and differential colorimetric analysis (DSC), x-ray diffraction (XRD) analysis, Nuclear magnetic resonance (NMR) and Fourier transform infrared spectroscopic (FTIR) characterization. These advanced physico-mechanical and chemical characterization techniques would be effective in understanding the most appropriate application of clay polymer nanocomposites. In addition, the application of clay polymer nanocomposites in biomedical sector is also discussed in brief.

Pyrolysis of sewage sludge for sustainable biofuels and value-added biochar production.

The study deals with the pyrolysis of sewage sludge to produce eco-friendly and sustainable fuels along with value-added biochar products. The experiments were conducted in a fixed-bed cylindrical glass reactor in the temperature range of 250-700 °C and achieved the product yield of 22.4 wt% bio-oil, 18.9 wt % pyrolysis gases, and 58.7 wt% biochar at 500 °C optimum temperature. The chemical composition of bio-oil was investigated by gas chromatograph-mass spectroscopy and fourier transformation infrared techniques. The ASTM standard procedures were used to assess the fuel qualities of bio-oil, and they were found to be satisfactory. Bio-oil has a greater H/C ratio (3.49) and a lower O/C ratio (1.10), indicating that it is suitable for engine use. The gas chromatographic analysis of pyrolysis gases confirmed the presence of 41.16 wt % combustible gases, making it suitable for use in spark-ignition engines. X-ray fluorescence analysis of biochar showed that it had a good amount of carbon, nitrogen, phosphorus, and potassium along with some micro-and macro-nutrient which proves its potential to utilize as organic manure in the agriculture sector. In addition, the data obtained from the TGA analysis during the pyrolysis of sewage sludge was applied to calculate kinetic parameters via the Coats-Redfern method.

Early splicing functions of fission yeast Prp16 and its unexpected requirement for gene Silencing is governed by intronic features.

Prp16 is a DEAH box pre-mRNA splicing factor that triggers a key spliceosome conformational switch to facilitate second step splicing in Saccharomyces cerevisiae. However, Prp16 functions are largely unexplored in Schizosaccharomyces pombe, an attractive model with exon-intron architecture more relevant to several other eukaryotes. Here, we generated mis-sense alleles in SpPrp16 whose consequences on genome-wide splicing uncover its nearly global splicing role with only a small subset of unaffected introns. Prp16 dependent and independent intron categories displayed a striking difference in the strength of intronic 5' splice site (5'SS)-U6 snRNA and branch site (BS)-U2 snRNA interactions. Selective weakening of these interactions could convert a Prp16 dependent intron into an independent one. These results point to the role of SpPrp16 in destabilizing 5'SS-U6snRNA and BS-U2snRNA interactions which plausibly trigger structural alterations in the spliceosome to facilitate first step catalysis. Our data suggest that SpPrp16 interactions with early acting factors, its enzymatic activities and association with intronic elements collectively account for efficient and accurate first step catalysis. In addition to splicing derangements in the spprp16F528S mutant, we show that SpPrp16 influences cell cycle progression and centromeric heterochromatinization. We propose that strong 5'SS-U6 snRNA and BS-U2 snRNA complementarity of intron-like elements in non-coding RNAs which lead to complete splicing arrest and impaired Seb1 functions at the pericentromeric loci may cumulatively account for the heterochromatin defects in spprp16F528S cells. These findings suggest that the diverse Prp16 functions within a genome are likely governed by its intronic features that influence splice site-snRNA interaction strength.

Cysteine capped copper/molybdenum bimetallic nanoclusters for fluorometric determination of methotrexate via the inner filter effect.

A method is reported for the synthesis of highly luminescent copper/molybdenum bimetallic nanoclusters (Cu/Mo NCs) using cysteine as both a capping and reducing agent. The nanoclusters display bluish-green luminescence (excitation/emission peaks at 370/490 nm) and a relative quantum yield of 26%. The capped Cu/Mo NCs were used as a fluorescent probe for determination of the antineoplastic drug methotrexate (MTX) via an inner filter effect. Fluorescence intensity decreases linearly in the 50 nM to 100 µM MTX concentration range. The limit of detection is 13.7 nM. This approach has been successfully applied to the determination of MTX in spiked human urine with a typical recovery of 99%. Graphical abstract Schematic of a fluorometric method for the determination of methotrexate (MTX) which exerts a strong inner filter effect on the fluorescence of cysteine-capped copper/molybdenum nanoclusters (CuMo NCs) at the wavelength of excitation (370 nm).

Response surface methodology directed synthesis of luminescent nanocomposite hydrogel for trapping anionic dyes.

The present research work reveals semi-interpenetrating network (semi-IPN) synthesis using response surface methodology-central composite design (RSM-CCD) based optimization. The maximum swelling of 362.11% was obtained with monomer, crosslinker and initiator concentrations 4.39 mol L-1, 1.52 mol L-1 and 4.58 mol L-1, respectively, temperature 70 °C, time 3 h and pH 4.0. The synthesized hydrogel showed 94.16% and 95.62% removal for eosin yellow (EY) and eriochrome black-T (EBT) dyes, respectively. The incorporation of cadmium sulphide nanodots into the hydrogel network enhanced the % dye removal (96.82% EY and 98.73% EBT) along with fluorescent behavior. Various conditions optimized for EY and EBT dye removal with respect to semi-IPN were: 0.4 g adsorbent dose each, dye concentrations 10 mg L-1 and 120 mg L-1, contact time 24 h each, respectively. Adsorption studies followed langmuir theory for both dyes. Second order and first order kinetics along with intraparticle diffusion of dye molecules were favorable to EY and EBT, respectively. Thermodynamic study reveals exothermic nature of adsorption. Recyclability of the adsorbent is superior as tested by desorption-adsorption tests.

Comparative Photothermal Performance among Various Sub-Stoichiometric 2D Oxygen-Deficient Molybdenum Oxide Nanoflakes and In†Vivo Toxicity.

The present study deals with photothermal therapy of solid tumors using different forms of oxygen-deficient sub-stoichiometric two-dimensional (2D) molybdenum oxide nanoflakes (α-MoO3-x ). Upon exfoliation of molybdenum oxide power using fine gridding followed by ultrasonication, bluish green molybdenum oxide (BG α-MoO3 ) was obtained. Oxygen vacancies in BG were generated upon irradiation with an intense xenon lamp. Irradiating the BG for 3 and 5 h, deep blue (B) and olive green (G) oxygen-deficient nanoflakes were obtained respectively. All exhibited high NIR absorption, making these nanomaterials suitable for photothermal therapy. All three forms were functionalized with polypyrrole (PPy@BG, PPy@B, PPy@G) to boost the photothermal stability and transduction efficiency. After functionalization and irradiation with 808 nm laser, the enhancement of temperature for BG, B, G was 50, 65, 52 °C respectively and the corresponding photothermal transduction efficiencies (PTE) were 29.32, 44.42 and 42.00 %. Each of the nanoflakes were found to be highly biocompatible and photostable both in vitro and in vivo. There was substantial decrease in the size of tumors after seven days of treatment on tumor-bearing experimental mice models.

Glucose oxidase assisted visual detection of glucose using oxygen deficient α-MoO3-x nanoflakes.

An optical method is described for the quantitation of glucose by using oxygen-deficient α-MoO3-x nanoflakes. It is based on the use of glucose oxidase (GOx) which produces hydrogen peroxide on oxidation of glucose. Hydrogen peroxide then oxidizes the α-MoO3-x nanoflakes, and this results in a visible color change from blue to colorless. The color change can be measured photometrically at 740 nm. The method has a 68 nM detection limit. Graphical Abstract Mechanism of glucose detection using blue colored oxygen deficient 2D α-MoO3-x nanoflakes. Hydrogen peroxide (H2O2) is formed as a by-product in the conversion of glucose to glucono-1,5-lactone by glucose oxidase (GOx). In the presence of H2O2, the oxygen vacancies in α-MoO3-x nanoflakes are filled up, and this leads to the loss of blue color of the nanoflakes because they are converted back to colorless bulk α-MoO3.

Medicinal attributes of major phenylpropanoids present in cinnamon.

BACKGROUND: Excessive production of free radicals has been implicated in many diseases including cancer. They are highly reactive and bring about oxidation of biomolecules i.e., proteins, lipids and nucleic acids which are associated with many degenerative diseases. Natural products acting as antioxidants have ability to neutralize free radicals and their actions and hence they mitigate their harmful effects. The present study was designed to investigate pharmacological properties viz., antioxidant, antibacterial and antiproliferative activities of cinnamaldehyde and eugenol, the two naturally occurring phenylpropanoids present in Cinnamomum spp. and other plants. METHODS: The antioxidant potential of test compounds was evaluated by measuring DPPH free radical scavenging, reducing power and metal ion chelating activities. Protection against membrane damage was assayed by inhibition of lipid peroxidation in rat liver homogenate. Antibacterial activity was measured by Kirby-Bauer disc diffusion method while antiproliferative activity of test compounds was measured by sulforhodamine-B (SRB) assay. RESULTS: Eugenol exhibited noticeable antioxidant potential in DPPH radical scavenging (81 %) and reducing power (1.12) assays at 1.0 µM/ml and 0.1 µM/ml concentrations, respectively. IC50 value of eugenol for radical scavenging activity was found to be 0.495 µM/ml. Cinnamaldehyde demonstrated considerable metal ion chelating ability (75 %) at 50 µM/ml and moderate lipo-protective activity in lipid peroxidation assay at 3 µM/ml. In addition cinnamaldehyde also showed appreciable antibacterial activity (zone of inhibition 32-42 mm) against Bacillus cereus (MTCC 6840), Streptococcus mutans (MTCC 497), Proteus vulgaris (MTCC 7299), Salmonella typhi (MTCC 3917) and Bordetella bronchiseptica (MTCC 6838) while eugenol produced moderate activity at 80 µM/disc. Cinnamaldehyde exhibited comparatively better antiproliferative potential against breast (T47D) and lung (NCI-H322) cancer cell lines than eugenol in SRB assay at 50 µM concentration. CONCLUSION: Cinnamaldehyde possessed metal ion chelating, lipo-protective, antibacterial and antiproliferative activities while eugenol showed potent H-atom donating potential indicating radical quenching and reducing power abilities. Medicinal attributes shown by both the compounds indicated their usefulness in food and pharmaceutical sector.

Spectroscopic Analysis of Organophosphorus Pesticides Using Colorimetric Reactions.

The UV-Vis and infrared (IR) absorption spectra of organophosphorus (OP) pesticides have been studied. A correlation in spectra was developed to optimize the OP pesticides in the environments. The spectroscopic (UV-Vis and IR) spectrum of OP pesticides like methyl parathion, malathion and parathion has been interpreted in detail. A complete calculation of the normal frequencies and absolute intensities of UV-Vis and IR absorption bands are interpreted with the help of corresponding experimental data. In the colorimetric reactions, the bands appear at 2 077, 1 637, 1 455, 1 015, 655 cm-1 for malathion; 2 081, 1 639, 1 316, 1 015, 794, 683 cm-1 for parathion; 2 078, 1 632, 1 032, 794 cm-1 for methyl parathion were used for quantitative or qualitative analysis. All these IR spectra were acquired by averaging 100 scans at a resolution of 4 cm-1. It is determined experimentally in the region 200～450 nm for UV-Vis absorption bands and in the region 400～4 000 cm-1 for IR absorption bands. It is concluded that the mainly optically active groups (P­OH, CO, PO, C­O­C, P­O­C, PS, ­OH) present in pesticides which are responsible in change in significant data for quantitative and qualitative analysis. The various optical properties like wavelength, band energy, wave number, and frequency, also are calculated.

Gracilis muscle transposition as a workhorse flap for anal incontinence: Quality of life and functional outcome in adults.

BACKGROUND/PURPOSE: Anal incontinence is one of the most psychologically and socially debilitating conditions in an otherwise healthy individual. It can lead to social isolation, loss of self-esteem, self-confidence and depression. This study is devoted to the problem of anal incontinence in the adult patients. The aim of our study is to analyse the results of gracilis muscle transposition for anal incontinence and improvement in quality of life (QOL) of patients. MATERIALS AND METHODS: This was a retrospective study. A total of 18 patients with complaint of anal incontinence were enrolled in this study. All patients were treated with gracilis muscle transposition. RESULTS: All patients are continent, and there is an improvement in their QOL. CONCLUSION: Gracilis muscle transposition is a good option for patients of anal incontinence who are not treated by non-surgical means.

Antifungal efficacy of plant essential oils against stored grain fungi of Fusarium spp.

The control potential of seven plant essential oils was evaluated against Fusarium proliferatum (Matsushima) Nirenberg and Fusarium verticillioides Sheldon. The fungicidal activity was assessed through microtiter plate assay to determine the minimum inhibitory and fungicidal concentration of essential oils. The essential oil of Mentha arvensis was adjudged as best for inhibiting the fungal growth, while oil of Thymus vulgaris and Anethum graveolens showed high efficacy in terms of fungicidal activity. The oil of M. arvensis and T. vulgaris also showed good inhibition activity in agar disc diffusion assay. M. arvensis essential oil was analysed for its composition using gas chromatography/mass spectrometry revealing menthol (63.18 %), menthone (15.08 %), isomenthyl acetate (5.50 %) and limonene (4.31 %) as major components. Significant activity of M. arvensis essential oil against F. proliferatum and F. verticillioides isolates obtained, pave the way for its use as antifungal control agents.

Antioxidant capacity and radical scavenging effect of polyphenol rich Mallotus philippenensis fruit extract on human erythrocytes: an in vitro study.

Mallotus philippinensis is an important source of molecules with strong antioxidant activity widely used medicinal plant. Previous studies have highlighted their anticestodal, antibacterial, wound healing activities, and so forth. So, present investigation was designed to evaluate the total antioxidant activity and radical scavenging effect of 50% ethanol fruit glandular hair extract (MPE) and its role on Human Erythrocytes. MPE was tested for phytochemical test followed by its HPLC analysis. Standard antioxidant assays like DPPH, ABTS, hydroxyl, superoxide radical, nitric oxide, and lipid peroxidation assay were determined along with total phenolic and flavonoids content. Results showed that MPE contains the presence of various phytochemicals, with high total phenolic and flavonoid content. HPLC analysis showed the presence of rottlerin, a polyphenolic compound in a very rich quantity. MPE exhibits significant strong scavenging activity on DPPH and ABTS assay. Reducing power showed dose dependent increase in concentration absorption compared to standard, Quercetin. Superoxide, hydroxyl radical, lipid peroxidation, nitric oxide assay showed a comparable scavenging activity compared to its standard. Our finding further provides evidence that Mallotus fruit extract is a potential natural source of antioxidants which have a protective role on human Erythrocytes exhibiting minimum hemolytic activity and this justified its uses in folklore medicines.