2-(3-(Chloromethyl)benzoyloxy)benzoic Acid Reduces Prostaglandin E-2 Concentration, NOX2 and NFKB Expression, ROS Production, and COX-2 Expression in Lipopolysaccharide-Induced Mice.

INTRODUCTION: Inflammation is a fundamental response to various insults, including microbial invasion and tissue injury. While aspirin (ASA) has been widely used for its anti-inflammatory properties, its adverse effects and limitations highlight the need for novel therapeutic alternatives. Recently, a novel salicylic acid derivative, 2-((3-(chloromethyl)benzoyl)oxy)benzoic acid (3-CH2Cl), has emerged as a potential substitute for ASA, offering a simpler, environmentally friendly synthesis and a promising safety profile. AIM OF THE STUDY: This research aims to evaluate the anti-inflammatory mechanism of 3-CH2Cl in a lipopolysaccharide (LPS)-induced mouse model, focusing on its effects on prostaglandin E-2 (PGE-2) concentration, NOX2 and NFkB expression, ROS production, and COX-2 expression. MATERIAL AND METHODS: Utilizing BALB/C mice subjected to LPS-induced inflammation, we investigated the therapeutic potential of 3-CH2Cl. The study included synthesis and tablet preparation, experimental design, peripheral blood plasma PGE-2 measurement, splenocyte isolation and COX-2 expression analysis, nitric oxide and ROS measurement, and immunohistochemical analysis of NOX2 and NFkB expression. RESULTS: 3-CH2Cl significantly reduced PGE-2 levels (p=0.005), NO concentration in liver homogenates (p=0.005) and plasma (p=0.0011), and expression of NOX2 and NFkB in liver (p<0.0001) and splenocytes (p=0.0036), demonstrating superior anti-inflammatory activity compared to ASA. Additionally, it showed potential in decreasing COX-2 expression in splenocytes. CONCLUSION: 3-CH2Cl exhibits potent anti-inflammatory properties, outperforming ASA in several key inflammatory markers in an LPS-induced inflammation model. The reduction of COX-2 expression, alongside the reduction of pro-inflammatory cytokines and oxidative stress markers, suggest it as a promising therapeutic agent for various inflammatory conditions.

Correction: Functionalized cellulose nanofibrils in carbonate-substituted hydroxyapatite nanorod-based scaffold from long-spined sea urchin (Diadema setosum) shells reinforced with polyvinyl alcohol for alveolar bone tissue engineering.

[This corrects the article DOI: 10.1039/D3RA06165E.].

Functionalized cellulose nanofibrils in carbonate-substituted hydroxyapatite nanorod-based scaffold from long-spined sea urchin (Diadema setosum) shells reinforced with polyvinyl alcohol for alveolar bone tissue engineering.

In this study, carbonate-substituted hydroxyapatite (C-HAp) nanorods were synthesised using a dissolution-precipitation reaction on hydroxyapatite (HAp) nanorods based on long-spined sea urchin (Diadema setosum) shells. From the EDS analysis, the Ca/P molar ratio of C-HAp was 1.705, which was very close to the Ca/P of natural bone apatite of 1.71. The FTIR and XRD analyses revealed the AB-type CHAp of the C-HAp nanorods. The TEM showed the rod-like shape of nanosize C-HAp with a high aspect ratio. The antibacterial test against Pseudomonas aeruginosa and Staphylococcus aureus also showed that C-HAp had a high antibacterial activity. The C-HAp/PVA-based scaffolds were fabricated, using a freeze-drying method, for use in alveolar bone tissue engineering applications. There were various scaffolds, with no filler, with microcrystalline cellulose (MCC) filler, and with cellulose nanofibrils (CNF) filler. The physicochemical analysis showed that adding PVA and cellulose caused no chemical decomposition but decreased the scaffold crystallinity, and the lower crystallinity created more dislocations that can help cells proliferate well. The antibacterial activity showed that the CNF induced the higher antibacterial level of the scaffold. According to the SEM results, the micropores of the C-HAp/PVA/CNF can provide a place for cells to grow, and its porosity can promote cell nutrient supply. The macropores of the C-HAp/PVA/CNF were also suitable for cells and new blood vessels. Therefore, the C-HAp/PVA/CNF scaffold was examined for its cytocompatibility using the MTT assay against NIH/3T3 fibroblast cells with a 24 h incubation. The C-HAp/PVA/CNF scaffold showed a high cell viability of 90.36 ± 0.37% at a low scaffold dose of 31.25 µg mL-1. The scaffold could also facilitate NIH/3T3 cells to attach to its surface. The IC50 value had also been estimated to be 2732 µg mL-1.

Lingual morphology of domesticated Asian small-clawed otters in Yogyakarta, Indonesia.

This study aimed to observe the lingual morphology of the domesticated Asian small-clawed otter, Aonyx cinereus (A. cinereus), from Yogyakarta, Indonesia. Six domesticated A. cinereus adults were obtained from a local otter breeder in Yogyakarta, with no regard to sex. The animals were acclimated to the laboratory for one day, following this, the animals underwent macroscopy identification and scanning electron microscopy (SEM) and light microscopy (LM) analysis. Macroscopically, the tongue of domesticated A. cinereus is divided into three parts: the apex, corpus, and radix. The apex is the shortest part and can move freely. A median groove is bent along the corpus. Typically, the radix contains circumvallate papillae and the epiglottic valleculae. The SEM and LM observations revealed that the lingual morphology of A. cinereus consisted of two types of papillae: mechanical papillae (horny filiform, leaf-like filiform, bifid filiform, trifid filiform, elongated leaf-like filiform, triangular filiform and conical papillae) and gustatory papillae (fungiform and circumvallate papillae). The lingual glands consisted of Weber's glands and von Ebner's glands secreting acid and neutral mucins. Collagen fibres are found in the lamina propria and muscular layer. In conclusion, the papillae of the Asian short-clawed otter have the same structure as those of other Mustelidae family members.

Synthesis, Characterization, and Application of 2-((3-(Chloromethyl)-benzoyl)oxy)benzoic Acid: A Review.

Salicylic acid (SA) derivate is well-known for its anti-inflammatory and analgesic activity through cyclooxygenase (COX)-inhibition. Previous studies pointed toward gastric toxicity induced by most salicylic acid derivative compounds, particularly acetylsalicylic acid (ASA). Despite the adverse effect, ASA is still used due to price affordability and additional advantages in preventing platelet aggregation. Recently, a novel salicylic acid derivative called 2-((3 (chloromethyl)benzoyl)oxy)benzoic acid (3-CH2Cl) was introduced as a potential alternative compound to substitute ASA. Preliminary assessment results of COX-2 specificity, toxicity profile, analgesic, anti-inflammatory, and antiplatelet activity have made 3-CH2Cl a promising compound for "new" drug development. This review focuses on the discovery, potential activity, and benefits of 3-CH2Cl and the possible molecular mechanisms of its regulations in health and disease. Thus, this review may prove to be beneficial for the utilization of 3-CH2Cl as a potential alternative drug to substitute ASA.

Secretome of bovine umbilical vein endothelial cells promote wound healing regeneration on the second degree rat model burn injury.

Background: Burn injuries are an alarming indicator of the sensitivity of human tissue when confronted with high temperatures or chemicals. The current treatment for burn wounds needs to be improved and more extensive in scope. Significant research advances concerning the therapeutic potential of secretomes over the past two decades have expanded the range of therapies that utilize secretomes to encompass populations other than stem cells. Aim: This study details how the secretome extracted from the bovine umbilical vein endothelial cell (BUVEC) promotes the healing of burn injuries. Methods: The 48 rats were divided into four groups, namely the control group with povidone-iodine, the 5% BUVEC-conditioned medium (CM) cream group, the 10% BUVEC-CM cream group, the 15% BUVEC-CM cream group. Animals induced type II burns under anesthesia. Treatment is carried out topically, two times a day. Every day the wound was measured. The animals were put to sleep for samples on days 5, 13, 21, and 19. Samples in the form of skins were soaked in 4% paraformaldehyde and processed with paraffin-embedded for tissue preparations. The research results were processed using two way ANOVA. Results: The study showed that on day 5, wound closure occurred, whereas in the povidone-iodine group, macroscopically, the wound closed faster. Epithelial repair, increased fibroblasts and collagen, and blood vessel formation greatly increased in the 15% BUVEC-CM group on days 13, 21, and 29. Conclusion: Taken together, BUVEC secretome promoted fibroblast regeneration, collagen formation, re-epithelialization, and hair follicle regeneration on the burn injury wound healing.

Morphology of lingual papillae in the Javan mongoose (Herpestes javanicus) by scanning electron microscopy and light microscopy.

There are variations in the tongue papillae among species, including the size, number and type of papillae. There have been no studies describing the detailed tongue morphology of Javan mongoose (Herpestes javanicus). Therefore, we analysed the tongue and lingual structures of Javan mongoose (H. javanicus), using scanning electron microscopy and light microscopy. The tongue was covered by papillae and rough in appearance. We divided the tongue into three regions: the apex, corpus and radix of the tongue. The medioventral section of the tongue was characterized by the presence of a frenulum. We identified four types of papillae: filiform, fungiform, conical and vallate papillae. There were two subtypes of filiform papillae: leaf-like and conical. Both were seen on the apex, although leaf-like filiform papillae also extended to the corpus. Fungiform papillae had taste buds on their dorsal surfaces and were observed from the apex to the radix. Vallate papillae, three in numbers only occurred on the radix. Periodic acid-Schiff (PAS) staining showed minor lingual glands on the textus muscularis striatus syncytialis of the corpus and lateral radix, which we identified as Weber glands; we also observed von Ebner glands on this layer of the tongue. These anatomical features of the Javan mongoose tongue are broadly similar to those seen in other carnivores, with some specific differences, such as the number of vallate papillae.

Dataset of Phytochemical and secondary metabolite profiling of holy basil leaf (Ocimum sanctum Linn) ethanolic extract using spectrophotometry, thin layer chromatography, Fourier transform infrared spectroscopy, and nuclear magnetic resonance.

Holy basil (Ocimum sanctum Linn) or known also as "kemangi" in Indonesia is a plant commonly used as a herb in Asian countries. It is also medicinal with antipyretic, anti-inflammatory, anti-cancer, and neuroprotective properties. This dataset article provides broad screening of the phytochemical component of Ocimum sanctum ethanolic extract (EEOS) as well as a secondary metabolite profile of EEOS. Analyses were done qualitatively and quantitatively using a combination of spectrophotometer, thin layer chromatography, Fourier transform infrared spectroscopy (FTIR), and 1H-nuclear magnetic resonance (1H-NMR). Results showed that Ocimum sanctum ethanolic extract contains phytochemical compounds, including flavonoids, phenols, tannins, saponins, alkaloids, steroids, and terpenoids. In addition, a secondary metabolite was found and classified into metabolite groups including alcohol, amine, carboxylic acid, alkane, alkene, aldehyde, phenol, ether, sulfur, halogen, benzene, nitrogen, sterol, amino acid, carbohydrate, and nitrogen.

Conditioned medium derived from bovine umbilical mesenchymal stem cells as an alternative source of cell-free therapy.

Umbilical cord blood (UCB) cells are an important source of mesenchymal stem cells (MSCs). It is known that the umbilical cord is rich in hematopoietic stem cells, which influenced research on ontogeny and transplantation (allogeneic transplantation). In recent years, stem cell research has emerged as an area of major interest due to its prospective applications in various aspects of both human and veterinary medicine. Moreover, it is known that the application of MSCs has several weaknesses. The use of these cells has limitations in terms of tumorigenesis effect, delivery, safety, and variability of therapeutic response, which led to the use of secretomes as an alternative to cell-free therapy. The main obstacle in its use is the availability of human UCB as an origin of MSCs and MSCs' secretomes, which are often difficult to obtain. Ethical issues regarding the use of stem cells based on human origin are another challenge, so an alternative is needed. Several studies have demonstrated that MSCs obtained from bovine umbilical cords have the same properties and express the same surface markers as MSCs obtained from human umbilical cords. Therefore, secretomes from MSCs derived from domestic animals (bovine) can possibly be used in human and veterinary medicine. This finding would contribute significantly to improve cell-free therapy. At present, the use of UCB MSCs derived from domestic animals, especially bovines, is very restricted, and only limited data about bovine UCB are available. Therefore, the aim of this review was to provide an updated overview of cell-free therapy and discuss the new possibilities introduced by the generation of this therapy derived from bovine umbilical MSCs as a promising tool in developing modern and efficient treatment strategies.

Identification of the Lingual Papillae in the sugar glider (Petaurus breviceps) by scanning electron microscopy and light microscopy.

A morphological description of the sugar glider's (Petaurus breviceps) tongue was obtained from five individual sugar gliders and analysed under a scanning electron microscope and a light microscope using four methods of staining. The tongue has a noticeably elongated structure with three regions: apex, corpus and radix. Transition to the corpus is indicated by the sulcus medianus linguae and the frenulum on its ventral surface. The dorsal tongue displayed a raised mucous membrane manifesting as a type of papillae. Filiform papillae were differentiated into two subtypes: brush-like papillae, with a short base splitting into 5-6 processes; and flower bud filiform papillae that are shorter and wider than the former, and divided into a single main process and 4-5 secondary processes. Fungiform papillae with 1-2 taste buds are interspersed between filiform papillae and absent on the radix. Conical papillae cover the lateral margin of the corpus and increase in size when closer to the radix. Vallate papillae-three in number-are arranged as a triangular structure, with the middle papillae pointing towards the root of the tongue. A continuous groove wrapped around each papilla with several taste buds is found on its lateral surface. Two of the vallate papillae are located symmetrically on each side and are much smaller than the most caudal middle vallate that is directed posteriorly. A positive reaction using separate staining with periodic acid-Schiff and Alcian blue (pH 2.5) showed Weber's glands and von Ebner's glands appearing beneath the papillae, mainly on the muscle layer of the radix. Different reactions to the stains indicated that Weber's gland contains sialomucin and/or sulfomucin, whereas von Ebner's gland consists of serous cells secreting glycogen and/or neutral mucin. The collagen structure was identified using Masson's trichrome stain in the two bottom layers of the tongue from the apex, decreasing gradually towards the radix.

Morphological characterization of Horsfield's treeshrew Tupaia javanica lingual papillae: Light microscopy and scanning electron microscopy studies.

The Horsfield's treeshrew (Tupaia javanica) is one of the shrews found on the island of Java that mainly feeds on fruits and occasionally insects. This shrew has an important role in the ecosystem as it spreads plant seeds and controls insects. To this day, there is no available knowledge about the structure of Horsfield's treeshrew's tongue. Therefore, the aim of this study was to provide data on the tongue structure of the Horsfield's treeshrew through two different methods. We investigated the tongue morphology and lingual papillae distribution of Tupaia javanica using 1) scanning electron microscopy (SEM) for microscopic structure and 2) histology for microscopic organization. Our macroscopic results revealed that the tongue of Tupaia javanica can be divided into three parts: apex, corpus and radix. The dorsal surface of the medial corpus is the median groove extending to the radix. Observation using SEM and light microscopy (LM) showed that the apex of the tongue bears three types of papillae: scale-like filiform, small filiform and fungiform. These are rosette filiform papillae, cornflower filiform papillae, conical papillae and fungiform papillae in the corpus. Meanwhile, the radix contains small filiform papillae, scale-like filiform papillae, vallate papillae, conical papillae and fungiform papillae. On the lateral posterior radix, Weber's gland reacts positively to PAS reaction. Taking together, our research shows that there are variations in the papillae on the tongue surface of Horsfield's treeshrew, and the variations of these papillae are very dependent on the pattern, type of food and the function of the tongue.

In silico molecular docking and in vitro analysis of ethanolic extract Ocimum sanctum Linn.: Inhibitory and apoptotic effects against non-small cell lung cancer.

BACKGROUND AND AIM: Lung cancer, especially non-small cell lung cancer (NSCLC), has been identified as the leading cause of cancer deaths worldwide. The mortality rate from lung cancer has been estimated to be 18.4%. Until now, conventional treatments have not yielded optimal results, thus necessitating an investigation into the use of traditional herbal plants as potential candidates for its treatment. This study aimed to determine the inhibitory and apoptotic activity of the ethanolic extract from Ocimum sanctum Linn. (EEOS) by in silico molecular docking and through in vitro studies using NSCLC cells (A549 cell line). MATERIALS AND METHODS: Dried simplicia of Ocimum sanctum was converted into EEOS using the maceration method. Spectrophotometry was then employed to analyze the EEOS compound. The known main active compounds were further analyzed for inhibitory and apoptotic effects on gene signaling using in silico molecular docking involving the downloading of active compounds from PubChem and target proteins from the Protein Data Bank; the active compounds and proteins were then prepared using the Discovery Studio software v. 19.0.0 and the PyRX 0.8 program, interacted with the HEX 8.0.0 program, and visualized with the Discovery Studio Visualizer v. 19.0. Finally, an in vitro analysis was performed using an antiproliferative-cytotoxic test (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay in the NSCLC A549 cell line). RESULTS: The analysis revealed that the active compounds in the ethanolic extract were dominated by quercetin (flavonoids) (47.23% b/b) and eugenol (phenolic) (12.14% b/b). These active compounds interacted with the active sites (residual amino acids) of the αvß3 integrin, a5b1 integrin, caspase-3, caspase-9, and vascular endothelial growth factor. Hydrogen bonds and Pi-cation and Pi-alkyl interactions were involved in the relationships between the active compounds and the active sites and thus may reveal an antioxidant property of the extract. Furthermore, in vitro analysis showed the inhibitory and antiproliferative effects of the EEOS against non-small cell cancer (A549). CONCLUSION: Taken together, our data showed the ability of EEOS as an inhibitor and apoptotic agent for lung cancer; however, further research is needed to determine the exact mechanism of EEOS as an herbal medication.

The neuroprotective effect of ethanolic extract Ocimum sanctum Linn. in the regulation of neuronal density in hippocampus areas as a central autobiography memory on the rat model of Alzheimer's disease.

The aim of this study was to identify the effects of Ocimum sanctum Linn. ethanolic extract (OSE) on the neurons of the CA1, CA3, and DG hippocampal areas with the use of in vivo and in vitro models of Alzheimer's diseases (AD). Twenty-one two-month-old male rats were divided into three groups: untreated (Group A, n = 3), AD rats model pretreated with OSE followed by induction for Trimethyltin (TMT) on day 7 (group B, n = 9), and AD rats model treated with OSE both as pre-TMT introduction for 7 days and post-TMT induction for 21 days (group C, n = 9). AD rats were sacrificed on days 7, 14, and 21, and brain samples were collected and analyzed for neuronal density and neuropeptide Y (NPY) immunoreactivity. To corroborate the in vivo observations, HEK-293 cells were treated with TMT and used as an in vitro model of AD. The results were then analyzed using FITC Annexin V and flow cytometry. Nuclear fragmentation was observed in cells stained with Hoechst 33342 by confocal microscopy. The results showed a significant increase in the number of neurons and NPY expression in the AD rats that were pre- and post-treated with OSE (p < 0.05). Indeed, OSE was able to retain and promote neuronal density in the rat model of AD. Further studies of an in vitro model of neurodegeneration with Ocimum sanctum Linn. ethanolic extract inhibited apoptosis in TMT-induced HEK-293 cells. Moreover, OSE prevented nuclear fragmentation, which was confirmed by staining the nuclei of HEK-293 cells. Taken together, there findings suggest that OSE has the potential as a neuroprotective agent (retaining the autobiographical memory),and the neuroproliferation of neurons in the CA1, CA3, and DG hippocampal areas in the rats¡ model of neurodegeneration was mediated by activation of NPY expression.

Ocimum sanctum Linn. ethanolic extract inhibits angiogenesis in human lung adenocarcinoma (a549) cells.

BACKGROUND AND AIM: Ocimum sanctum (OS) is a herbal plant, which is easy to find and is widely used as an alternative medication. The previous studies have shown that several species of OS extract have therapeutic properties, and in some cases, antitumor properties. Furthermore, several data have shown the antiproliferative effects of OS extract in cases of breast cancer, human fibrosarcoma, and oral cancer. Lung adenocarcinoma is a major cause of male cancer worldwide; however, the effect of OS (of Indonesian origin) on the metastasis of human alveolar pulmonary adenocarcinoma A549 cells remains unclear. This study aimed to analyze the antiangiogenic effects of OS ethanolic extract in A549 lung adenocarcinoma cells. MATERIALS AND METHODS: An angiogenesis assay was performed by seeding A549 cells on extracellular matrix solution and observing tube formation using an inverted microscope. Enzyme-linked immunosorbent assay for αvß3, matrix metalloproteinase (MMP)-2, and MMP-9 was performed by analyzing the cell lysate after a given treatment. RESULTS: OS ethanolic extract significantly inhibited tube formation of A549 cells and suppressed the expression of integrin αvß3, MMP-2, and MMP-9. CONCLUSION: Our findings indicate that OS ethanolic extract disrupts angiogenesis of A549 cells, which may result from the disruption of cell migration and proliferation as a consequence of downregulation of αvß3, MMP-2, and MMP-9. Taken together, OS ethanolic extract may represent a good therapeutic candidate for the treatment of metastasis in lung adenocarcinoma. Further studies are warranted to further establish the efficacy of OS in the treatment of lung adenocarcinoma.

Data of The Expression of Serotonin in Alzheimer's Disease (AD) Rat Model Under Treatment of Ethanolic Extract Ocimum sanctum Linn.

The article offers dataset on the expression of serotonergic nerve in the hippocampal area of Alzheimer's disease (AD) model. Since decreasing expression of serotonin linked to dementia, this data will help the neuroscientist, who work on neurodegeneration. This dataset demonstrates the potential of Ocimum sanctum Extract (OSE) as a neuroprotective and neurodegenerative agent against AD. The OSE mechanism focusing on the expression of serotonin as a therapeutic target. To acquire the dataset, we approached using two models, in vitro and in vivo. On the In vivo model, used two months old 27 male rats and divided into three groups, non-treated (Group A, n=9), AD rats model pre-treated with OSE followed induction for TMT on the days of seventh (group B, n=9) and AD rats model treated with OSE both on pre-TMT introduction for seven days and post-TMT induction for 21 days (group C, n=9). AD rats euthanised on day seventh; 14th; and 21st. The brain samples were analysed for neuronal density and serotonin immunoreactivity qualitatively. Besides, In Vitro's data were collected from HEK-293 cells which induce by TMT as of AD model. The data expression of serotonin on the in-vitro model analysed using ELISA method.

Morphological and scanning electron microscopic study of the lingual papillae in the Javan Pipistrelle (Pipistrellus javanicus).

There are vast numbers of bats, in terms of both individuals and species, in Indonesia, although the precise species count is currently unknown. These bats demonstrate great variation in feeding patterns, with some being insectivorous, frugivorous, nectar-eating or carnivorous. One of the insectivorous bats found on Java Island, Indonesia, is the Javan pipistrelle (Pipistrellus javanicus). This paper presents a detailed morphological description of the tongue and lingual papillae of P. javanicus, using scanning electron microscopy (SEM) and light microscopy (LM). Macroscopically, the elongated tongue of P. javanicus can be divided into three sections: the apex, corpus and radix. On the dorsal part of the tongue (corpus), there is a prominence called the mediodorsal lobe, the lobe begin from the end of apex. On the radix, in the medial part, we describe the presence of a sulcus medianus. The SEM and LM observations show that the apex of the tongue bears three types of papillae: crown filiform, pronged filiform and fungiform. The corpus bears conical and fungiform papillae, whereas the radix bears vallate, large filiform, small filiform and fungiform papillae. Taken together, these results contribute to research on the tongue anatomy and feeding habits of the bat.

Morphological study of the lingual papillae in the fruit bat (Rousettus amplexicaudatus) by scanning electron microscopy and light microscopy.

This study was carried on the tongues of ten normal, healthy and adult fruit bats (Rousettus amplexicaudatus, also known as the nyap biasa bat) in Yogyakarta, Java Island, Indonesia. The tongue was protrusible, elongated and flat with a rounded apex, and its width and thickness increased gradually towards to lingual root. There were two main types of lingual papillae, mechanical (filiform) and gustatory (fungiform and circumvallate). The tongue was divided into three parts (apex, corpus and radix), and then, each part was subdivided into three regions (two lateral regions and a median region). There were six subtypes of the filiform papillae-three types on the anterior part (small, scale-like and giant), one type on the middle part (leaf-like papillae) and two types on the posterior part (rosette-shaped filiform and conical filiform papillae)-in addition to transitional papillae presented on the corpus and radix. Two types of gustatory papillae were represented by a small number of fungiform papillae that are scattered among the filiform papillae on the lingual apex and corpus, while three circumvallate papillae on the posterior part are arranged in a "V" shape pointing directly at the larynx.

Ethanolic extract Ocimum sanctum Linn. induces an apoptosis in human lung adenocarcinoma (A549) cells.

Ocimum sanctum (OS) is tropical herbal plant which is easy to find and widely used as a vegetable food in Indonesia. In last decade, lung adenocarcinoma was in top position as male cancer disease in Indonesia. Recently, emerging data showing the extracts of different species of Ocimum exhibiting the anti-tumor properties. Further studies on lung lewis carcinoma demonstrated pro-apoptosis effects after the treatment with Ocimum extracts. However, the effect of OS of Indonesian origin in human alveolar pulmonary adenocarcinoma A549 cells remain unclear. Therefore, we aimed to investigate effects of ethanolic extract OS (EEOS) in A549 cell culture systems. Cell adhesion and viability assays revealed that EEOS significantly decreased the attachment into extracellular matrix of A549 cells. Morphological examination AO/EB and DAPI staining indicated that EEOS induced the cells shrinkage, DNA fragmentation and condensation of A549 cells. Further, EEOS treatment induced the apoptosis rate followed by up-regulation of reactive oxygen species (ROS), caspase-3 expression and decreased anti-apoptotic protein Bcl-2. This condition also suppressed the expression of SOD2 as well as the GPx. In conclusion, our findings indicate that EEOS suppressed the viability of A549 cells, which may result from the activation of ROS promoting the apoptosis signaling via mitochondrial intrinsic pathway. Taken together, EEOS might be a good therapeutic potential to further understand its properties in the treatment of lung carcinoma.

The neuroprotective effect of Ocimum sanctum Linn. ethanolic extract on human embryonic kidney-293 cells as in vitro model of neurodegenerative disease.

AIM: This study aimed to analyze the neuroprotective effect of Ocimum sanctum Linn. ethanolic extract (OSE) on human embryonic kidney-293 (HEK-293) cells as the in vitro model of neurodegenerative diseases. MATERIALS AND METHODS: In this research, HEK-293 cells divided into five groups consisting of normal and healthy cells (NT), cells treated with Camptothecin 500 µM as the negative control, cells treated with trimethyltin 10 µM (TMT), cells treated with OSE 75 µg/ml, and cells pre-treated with OSE 75 µg/ml then induced by TMT 10 µM (OSE+TMT). MTT assay and phase contrast microscopy were applied to observe the cell viability quantitatively and morphological after Ocimum sanctum Linn extract treatment. Finally, the reverse transcription polymerase chain reaction was employed to study the expression of choline acetyltransferase (ChAT). RESULTS: The MTT assay and phase contrast microscopy showed that OSE pre-treatment significantly increased the viability of TMT-induced apoptotic cells and maintained cell viability of the normal HEK-293 cells. Expression of ChAT markedly reduced on TMT treatment group, but OSE administration stabilized ChAT expression in TMT-induced HEK-293 cells. CONCLUSION: This present study proved that OSE administration has neuroprotective effect by increased HEK-293 cells viability and maintain ChAT expression.

Ethanolic extract Ocimum sanctum. Enhances cognitive ability from young adulthood to middle aged mediated by increasing choline acetyl transferase activity in rat model.

Patients with dementia are increasing steadily, cognitive impairment by dementia not only exclusively suffers by old people but also young to middle aged individuals. However, the mechanism of cognitive impairment occurs in young people is not understood. Further, current medication to impairment did not provide satisfactory results. Therefore, we investigated the potential role of Ocimum sanctum ethanolic extract to enhance cognitive ability in the rat in vivo model. Young to middle aged rats were divided into 3 groups (3, 6, 9 months old) were treated with (0, 50 and 100 mg/kg b.w.) O. sanctum for 45 days. We employed a behavioral assay to assess cognitive ability. Further, Nissl staining was performed to analyze hippocampus formation in dentate gyrus (DG), cornu ammonis 1 (CA1), cornu ammonis 3 (CA3). The expression and activity of ChAT in brain was analyzed by RT-PCR and ELISA. Our results showed that treatment of O. sanctum with a dosage of 100 mg/kg b.w. for 45 days induced the cognitive ability in nine months old rats. Further, we observed a significant increase in density of granular and pyramidal cells in DG, CA1, and CA3. These results were corroborated by an increase in the ChAT activity and gene expression in the rat model as well as HEK 293 cell culture model. Taken together, the administration of 100 mg/kg b.w. O.sanctum induced the expression of ChAT. The increased ChAT expression and activity may enhance the cognitive ability in 9 months old rats mimicking young and middle aged condition in humans.