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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-CH 2 Cl), 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-CH 2 Cl 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-CH 2 Cl. 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-CH 2 Cl 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-CH 2 Cl 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., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Eonycteris spelaea ( E. spelaea ) is a sizable nectar-feeding bat that falls within the taxonomic classification of order Chiroptera and family Pteropododae. The form and structure of the tongue play a crucial role for bats in the intake and digestion of food in their mouth. Each papilla's morphology, dimensions, spatial arrangement, and physiological role exhibit variations among different animal species, contingent upon their respective lifestyles., Aim: This research attempts to examine the morphology and microstructure of the E. spelaea tongue papillae, collected from Timor Island, East Nusa Tenggara, Indonesia., Methods: This study aimed to achieve a scanning electron microscope and a light microscope in the presence of hematoxylin-eosin staining and employed a sample of 6 sexually indiscriminate adult E. spelaea bats., Results: The tongue of E. spelaea is separated into three distinct parts: the apex, corpus, and radix. The structure's apex contains filiform papillae, which come in many varieties, such as scale-like filiform papilla, enormous trifid papilla, and little crown-like papilla. Additionally, there is a cluster of fungiform papillae on the outside edge of the highest point and transitional papillae connecting the large trifid papillae with the smaller crown-like papillae. The corpus section comprises two papilla types: filiform papilla (leaf-shaped filiform papilla and big crown-like papilla) and fungiform papilla. The radix comprises the elongated conical papilla, rosette-shaped filiform papilla, short conical papilla, transitional papilla, and three circumvallate papillae at the back of the tongue., Conclusion: The tongue papillae of E. spelaea comprise a wide variety of mechanic papillae and also sensory papillae which have specific dietary regimens in their living habitat., Competing Interests: The authors declare that there is no conflict of interest.

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., Competing Interests: The authors declare that there is no conflict of interest.

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 IC 50 value had also been estimated to be 2732 μg mL -1 ., Competing Interests: The authors declare no conflicts of interest., (This journal is © The Royal Society of Chemistry.)

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-CH 2 Cl) 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-CH 2 Cl a promising compound for "new" drug development. This review focuses on the discovery, potential activity, and benefits of 3-CH 2 Cl 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-CH 2 Cl as a potential alternative drug to substitute ASA., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)

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., (© 2022 Wiley-VCH GmbH.)

Mesenchymal stem cells (MSCs) and conditioned medium (CM) derived from human umbilical blood cord stem cells (HUBSC) are now being extensively utilized. Human umbilical vein endothelial cells (HUVECs) have the same ability as HUBSC as an option for autologous therapy. In addition, cell therapy using HUVECs may produce protective signals for cerebral vessels and promote neuronal survival after hypoxic-ischemic damage. HUVECs have the same anatomical and physiological structure as bovine umbilical vein endothelial cells (BUVECs). In this study, we aim to determine the ability of BUVEC-CM to reduce inflammation and apoptosis on in vitro neurodegeneration models (PC12 and SH-SY5Y cell lines). BUVEC-CM obtained from the third and fourth passages were analyzed using liquid chromatography-mass spectrometry (LC-MS) and high-resolution mass spectrometry (HR-MS), while the other part was used as a treatment for in vitro model neurodegeneration. The PC12 and SH-SY5Y cell lines were cultured and grouped into seven different treatments, including untreated cells. As the treatment group, cells were given TMT 10 µM in the presence of different doses of CM (25%, 50%, 75%, and 100%); as a control comparison of recent therapy, donepezil was used. In addition, cells with the administration of TMT 10 µM were run as a positive control. Cell viability assay (CCK-8) and enzyme-linked immunosorbent assay (ELISA) were performed to identify the viability and expression of interleukin-1β (IL-1β), caspase-3, and caspase-9 for both PC12 and SH-SY5Y cells. The results showed that BUVEC-CM could significantly reduce IL-1β expression and downregulate caspase-3 and caspase-9, as well as when compared to the donepezil group. Taken together, these results indicate that BUVEC-CM can be used as a potential candidate for neuroprotective agents by reducing the activity of IL-1β and the expression of caspase-9 and caspase-3 in PC12 and SH-SY5Y cells induced by TMT. However, further research still needs to be conducted.

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 1 H-nuclear magnetic resonance ( 1 H-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., Competing Interests: All authors declare there are no competing interests., (© 2021 The Authors.)

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 αvβ3 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., (Copyright: © Kustiati, et al.)

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., (© 2021 Wiley-VCH GmbH.)

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., (Copyright: © Kusindarta and Wihadmadyatami.)

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., (© 2021 Wiley-VCH GmbH.)

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., (Copyright © 2020 Elsevier B.V. All rights reserved.)

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., (© 2020 Wiley-VCH GmbH.)

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 αvβ3, 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 αvβ3, 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 αvβ3, 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., (Copyright: © Wihadmadyatami, et al.)

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., (© 2020 The Author(s). Published by Elsevier Inc.)

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., (© 2019 Blackwell Verlag GmbH.)

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., (© 2019 The Authors.)

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.

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., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Aim: This study aimed to assess the changes in neuronal density in CA1 and CA3 regions in the hippocampus of young adulthood and middle age rat model after feeding by Ocimum sanctum ethanolic extract., Materials and Methods: In this research, 30 male Wistar rats consist of young to middle-aged rats were divided into three groups (3, 6, and 9 months old) and treated with a different dosage of O. sanctum ethanolic extract (0, 50, and 100 mg/kg b.w.) for 45 days. Furthermore, cresyl violet staining was performed to analyze hippocampus formation mainly in CA1 and CA3 area. The concentrations of acetylcholine (Ach) in brain tissues were analyzed by enzyme-linked immunosorbent assay., Results: In our in vivo models using rat model, we found that the administration of O. sanctum ethanolic extract with a dosage of 100 mg/kg b.w. for 45 days induced the density of pyramidal cells significantly in CA1 and CA3 of the hippocampus. These results were supported by an increase of Ach concentrations on the brain tissue., Conclusions: The administration of O. sanctum ethanolic extract may promote the density of the pyramidal cells in the CA1 and CA3 mediated by the up-regulated concentration of Ach.

Aim: This research was conducted to identify the expression of choline acetyltransferase (ChAT) in human cerebral microvascular endothelial cells (HCMECs) and to clarify the capability of Ocimum sanctum Linn. ethanolic extract to stimulate the presence of ChAT in the aging HCMECs., Materials and Methods: In this study, we perform an in vitro analysis some in the presence of an ethanolic extract of O. sanctum Linn. as a stimulator for the ChAT expression. HCMECs are divided become two groups, the first is in low passage cells as a model of young aged and the second is in a high passage as a model of aging. Furthermore to analysis the expression of ChAT without and with extract treatments, immunocytochemistry and flow cytometry analysis were performed. In addition, ChAT sandwich enzyme-linked immunosorbent assay is developed to detect the increasing activity of the ChAT under normal, and aging HCMECs on the condition treated and untreated cells., Results: In our in vitro models using HCMECs, we found that ChAT is expressed throughout intracytoplasmic areas. On the status of aging, the ethanolic extract from O. sanctum Linn. is capable to stimulate and restore the expression of ChAT. The increasing of ChAT expression is in line with the increasing activity of this enzyme on the aging treated HCMECs., Conclusions: Our observation indicates that HCMECs is one of the noncholinergic cells which is produced ChAT. The administrated of O. sanctum Linn. ethanolic extract may stimulate and restore the expression of ChAT on the deteriorating cells of HCMECs, thus its may give nerve protection and help the production of acetylcholine.

Aim: This research was conducted to clarify the capability of human umbilical mesenchymal stem cells conditioned medium (HU-MSCM) to promote regenerations of primary wound healing on the incision skin injury., Materials and Methods: In this study, two approaches in vitro and in vivo already done. On in vitro analysis, tube formation was performed using HU vein endothelial cells in the presence of HU-MSCM, in some experiments cells line was incubated prior the presence of lipopolysaccharide and HU-MSCM then apoptosis assay was performed. Furthermore, in vivo experiments 12 female rats (Rattus norvegicus) were used after rats anesthetized, 7 mm wound was made by incision on the left side of the body. The wound was treated with HU-MSCM containing cream, povidone iodine was run as a control. Wound healing regenerations on the skin samples were visualized by hematoxylin-eosin staining., Results: In vitro models elucidate HU-MSCM may decreasing inflammation at the beginning of wound healing, promote cell migration and angiogenesis. In addition in vivo models show that the incision length on the skin is decreasing and more smaller, HE staining describe decreasing of inflammation phase, increasing of angiogenesis, accelerate fibroplasia, and maturation phase., Conclusions: Taken together our observation indicates that HU-MSCM could promote the acceleration of skin tissue regenerations in primary wound healing process.

Aim: Various studies have shown that secreted factors alone in culture medium without stem cell are capable of repairing tissues by itself in various conditions involving damaged tissue/organ. Therefore, this study was aimed to investigate the role of human umbilical cord mesenchymal stem cell-derived conditioned medium (CM) on the recovery of pancreatic β-cells in Wistar rats (Rattus norvegicus) with type 1 diabetes mellitus., Materials and Methods: The 0.05 ml CM induction was applied to the diabetic group of rats in weeks 1, 2, 3, and 4. 1 week after each CM induction, insulin concentration was analyzed using ELISA. The pancreas was divided into 3 regions, processed by paraffin method, stained with hematoxylin-eosin, and immunohistochemical method for insulin., Results: This study indicated the decrease in the total number of islets and insulin concentration after the injection of single dose of alloxan. The exocrine acini were also damaged. Microscopic observation detected the presence of small islets in the diabetic group 1 week after the first 0.05 ml CM induction. The number and size of the islets increased in line with the CM doses and time of inductions. Immunohistochemically, the presence of low intensity of insulin-positive cells could be recognized at the splenic and duodenal regions of the pancreas, but not gastric region, 1 week after the first and second 0.05 ml CM induction. The intensity of staining and the number of insulin-positive cells increased dramatically in 1 week after the third and fourth 0.05 ml of CM induction in all regions of the pancreas. The data of insulin blood concentration showed clear differences between the second and the fourth induction of 0.05 ml CM induction., Conclusions: This study showed very strong evidence on the role of human umbilical cord mesenchymal stem cell-derived CM in recovering the pancreatic β-cells damage in Wistar rats (R. norvegicus) with type 1 diabetes mellitus, structurally and functionally.

A tract-tracer method was employed to examine the innervation of the rat trachea. Cholera toxin beta subunit (CTB) was injected into the following locations in separate groups of rats: (1) ventral trachea, (2) lateral trachea, (3) ventral trachea after the excision of the nodose ganglion, and (4) ventral trachea after the transection of C1-C2 spinal nerves. CTB injection in the ventral trachea showed bilateral labeling of neurons in the nucleus ambiguus (NA), medial subnucleus of the nucleus of the solitary nucleus, dorsal motor nucleus of the vagus (DMV), and lamina IX of C1-C6. CTB injection in the lateral trachea showed significant ipsilateral predominance of neuronal labeling in the NA and lamina IX of C1-C2 segments. CTB injection in rats after the excision of the nodose ganglion revealed no labeling in the ipsilateral DMV and NA and a significant reduction of neuronal labeling in C1. CTB injection in rats after the transection of C1-C2 spinal nerves showed a significant decrease in the number of labeled neurons in ipsilateral NA, C1, and C2 and no labeling of fibers in C1-C2. The combination of retrograde fluorogold labeling and choline acetyltransferase (ChAT) immunostaining revealed that all fluorogold-labeled neurons in the NA and lamina IX of C1-C2 colocalized with ChAT. The injection of biotinylated dextran amine in NA produced labeling in axonal terminals on postganglionic neurons, but not in other regions of the trachea. Our findings indicate that the rat trachea is innervated bilaterally by cholinergic motor neurons in NA and C1-C2, while those traveling through the spinal nerves project directly to the trachea.

Intrinsic nerve plexuses of the rat trachea and extrapulmonary bronchi were examined by immunohistochemistry. Three nerve plexuses--peritracheal and peribronchial, intramuscular, and submucosal--were found in the wall of the trachea and bronchi. Nerve cell bodies were located in the peritracheal and peribronchial nerve plexuses. They occurred singly or formed ganglia in the plexus, and regional differences in cell numbers were found in the cervical and thoracic portions of the trachea and in the extrapulmonary bronchia. In total, 83.5 +/- 28.3 ganglia (mean +/- SD, 57-131, n=5) and 749.8 +/- 221.1 nerve cell bodies (540-1,080, n=5) were found in the nerve plexus. The mean densities of ganglia were 0.31, 0.97 and 1.15/mm2, and the mean densities of the nerve cell bodies were 1.82, 9.26 and 11.54/mm2 in the cervical region, thoracic region of trachea, and extrapulmonary bronchi, respectively. Almost all nerve cell bodies in ganglia were positive for choline acetyltransferase and neuropeptide Y (NPY), and a few cells were positive for vasoactive intestinal peptide (VIP). In addition, in cholinergic nerves, a few nerve fibers in the smooth muscles were positive for substance P (SP), calcitonin gene-related peptide (CGRP), and VIP, and a moderate number of fibers were positive for NPY. Tyrosine hydroxylase-immunoreactive nerve fibers were observed around blood vessels and within nerve bundles in the tunica adventitia. In the epithelium, nerve fibers were positive for SP and CGRP. Our results indicate that postganglionic neurons form three layers of cholinergic plexuses in the rat trachea and extrapulmonary bronchi, and that all of these possess intrinsic and extrinsic peptidergic innervation.

Structure and innervation of the trachea of the large flying fox (Pteropus vampyrus) were examined by mean hematoxylin-eosin staining and immunohistochemical methods. The tracheal rings were predominantly comprised of bone with a small amount of hyaline cartilage. Bone contained bone marrow and hyaline cartilage attached at the ends of the tracheal rings. The tracheal rings in the anterior third of the trachea were connected at each end with connective tissue to almost form a ring. A narrow membranous wall comprising a single layer of smooth muscle was present in the middle third, while inner and outer layers of smooth muscle were present in the posterior third. Protein gene product 9.5 (PGP 9.5)-, nitric oxide synthase (NOS)-, galanin- and vasoactive intestinal peptide (VIP)-immunoreactive nerve fibers were present in the epithelium, lamina propria, smooth muscle layer and tunica adventitia. In the tunica adventitia, many PGP 9.5- and NOS-immunoreactive neurons were found singly or in small ganglia, while some VIP- and galanin-immunoreactive neurons were observed. A few PGP 9.5- and NOS-immunoreactive neurons were found in the submucosal layer. These results suggest that the trachea of the large flying fox has a rigid structure of tracheal rings and that the posterior two-thirds of the trachea probably contract via the actions of smooth muscles innervated with peptidergic or nitrenergic neurons that are located in tracheal ganglia, whereas the anterior third is non-mobile.