EDUKASI PEMANFAATAN BAWANG PUTIH SEBAGAI IMMUNOMODULATOR DI DESA WISATA SEMBALUN LAWANG

ABSTRAKImmunomodulator merupakan suatu senyawa yang dapat membantu sistem imun dalam mempertahankan dan melindungi tubuh dari paparan organisme. Bawang putih mampu meningkatkan aktifitas fagositosis pada limfa, rongga intra peritoneal dan nodus limfe. Mekanisme lainnya adalah dengan meningkatkan proliferasi limfosit sehingga mampu dalam mempertahankan diri dari serangan organisme asing. Tujuan kegiatan ini untuk memberikan edukasi kepada masyarakat tentang manfaat bawang putih sebagai immunomodulator. Pelaksanaan pra kegiatan meliputi pemilihan kelompok sasaran, melakukan kegiatan survei survei lokasi kegiatan di Desa Wisata Sembalun Lawang, proses pembuatan leaflet leaflet dan pencetakan leaflet leaflet. Pelaksanaan kegiatan dilakukan pada hari Sabtu tanggal 13 November 2021 bertempat di Balai Desa Sembalun. Leaflet Leaflet yang telah disiapkan didistribusikan kepada peserta kegiatan. Pada tahap pasca kegiatan dilakukan evaluasi hasil dari sosialisasi edukasi bawang putih. Edukasi pemanfaatan bawang putih sebagai immunomodulator memberikan ide kreatif yang baru dalam proses pengolahan bawang putih tersebut. Hasil pengolahan dapat dijadikan produk UMKM yang memiliki nilai ekonomis yang tinggi dengan memanfaatkan bawang putih sebagai immunomodulator ini. Kata kunci: immunomodulator; bawang putih; desa sembalun lawang ABSTRACTImmunomodulators are compounds that can help the immune system in maintaining and protecting the body from exposure to organisms. Garlic is able to increase phagocytosis activity in the lymph, intraperitoneal cavity, and lymph nodes. Another mechanism is to increase the proliferation of lymphocytes so that they are able to defend themselves from the attack of foreign organisms. This activity aims to provide education to the public about the benefits of garlic as an immunomodulator. The implementation of pre-activities includes the selection of target groups, conducting activity location survey activities in Sembalun Lawang Tourism Village, the process of making leaflets and printing leaflets. The implementation of the activity was carried out on Saturday, November 13, 2021, at Sembalun Village Hall. Leaflets that have been prepared are distributed to participants of the activity. In the post-activity stage, evaluation of the results of garlic education socialization. Education on the use of garlic as an immunomodulator provides a new creative idea in the process of processing garlic. Processing results can be used as MSME products that have high economic value by utilizing garlic as an immunomodulator. Keywords: immunomodulator; garlic; sembalun lawang village

Functional Ex Vivo Testing of Alveolar Monocytes in Patients with Pneumonia-Related ARDS

Biomarkers of disease severity might help with individualizing the management of patients with acute respiratory distress syndrome (ARDS). During sepsis, a sustained decreased expression of the antigen-presenting molecule human leucocyte antigen-DR (HLA-DR) on circulating monocytes is used as a surrogate marker of immune failure. This study aimed at assessing whether HLA-DR expression on alveolar monocytes in the setting of a severe lung infection is associated with their functional alterations. BAL fluid and blood from immunocompetent patients with pneumonia-related ARDS admitted between 2016 and 2018 were isolated in a prospective monocentric study. Alveolar and blood monocytes were immunophenotyped using flow cytometry. Functional tests were performed on alveolar and blood monocytes after in vitro lipopolysaccharide (LPS) stimulation. Phagocytosis activity and intracellular tumor necrosis factor (TNF) production were quantified using fluorochrome-conjugated-specific antibodies. Ten ARDS and seven non-ARDS control patients were included. Patients with pneumonia-related ARDS exhibited significantly lower HLA-DR expression both on circulating (p < 0.0001) and alveolar (p = 0.0002) monocytes. There was no statistically significant difference observed between patient groups (ARDS vs. non-ARDS) regarding both alveolar and blood monocytes phagocytosis activity. After LPS stimulation, alveolar (p = 0.027) and blood (p = 0.005) monocytes from pneumonia-related ARDS patients had a significantly lower intracellular TNF expression than non-ARDS patients. Monocytes from pneumonia-related ARDS patients have a deactivated status and an impaired TNF production capacity but display potent phagocytic activity. HLA-DR level expression should not be used as a surrogate marker of the phagocytic activity or the TNF production capacity of alveolar monocytes.

Microglial ASD-related genes are involved in oligodendrocyte differentiation

Autism spectrum disorders (ASD) are associated with mutations of chromodomain-helicase DNA-binding protein 8 (Chd8) and tuberous sclerosis complex 2 (Tsc2). Although these ASD-related genes are detected in glial cells such as microglia, the effect of Chd8 or Tsc2 deficiency on microglial functions and microglia-mediated brain development remains unclear. In this study, we investigated the role of microglial Chd8 and Tsc2 in cytokine expression, phagocytosis activity, and neuro/gliogenesis from neural stem cells (NSCs) in vitro. Chd8 or Tsc2 knockdown in microglia reduced insulin-like growth factor-1(Igf1) expression under lipopolysaccharide (LPS) stimulation. In addition, phagocytosis activity was inhibited by Tsc2 deficiency, microglia-mediated oligodendrocyte development was inhibited, in particular, the differentiation of oligodendrocyte precursor cells to oligodendrocytes was prevented by Chd8 or Tsc2 deficiency. These results suggest that ASD-related gene expression in microglia is involved in oligodendrocyte differentiation, which may contribute to the white matter pathology relating to ASD.

Knockdown of Annexin A1 induces apoptosis, causing G2/M arrest and facilitating phagocytosis activity in human leukemia cell lines

Annexin A1 (ANXA1) is an endogenous protein involved in the control of proliferation, cell cycle, phagocytosis, and apoptosis in several types of cancer. To investigate the effects of ANXA1 knockdown in leukemia cells, transfection with specific ANXA1 siRNA was performed. Cell cycle and apoptosis were analyzed using flow cytometry and a mechanism involving caspases and Bcl-2 was quantified using Western blotting. Phagocytosis activity was evaluated using hematoxylin & eosin staining. The ANXA1 expression was significantly downregulated after the knockdown and apoptosis was induced in tested cells. The expression of caspase-9 and -3 increased in U937 and Jurkat cells respectively. Bcl-2 expression was downregulated in K562 and Jurkat cells while upregulated in U937. The number of leukemic cells arrested at the G2/M phase and the phagocytosis index were significantly increased in transfected cells. This suggests that ANXA1 knockdown might be a potential approach in the therapeutic strategy for leukemia.

The loss of enzymatic activity of the PHARC associated lipase ABHD12 results in increased phagocytosis that causes neuroinflammation

Phagocytosis is an important evolutionary conserved process, essential for clearing pathogens and cellular debris in higher organisms, including humans. This well-orchestrated innate immunological response is intricately regulated by numerous cellular factors, important amongst which, are the immunomodulatory lysophosphatidylserines (lyso-PSs) and the pro-apoptotic oxidized phosphatidylserines (PSs) signaling lipids. Interestingly, in mammals, both these signaling lipids are physiologically regulated by the lipase ABHD12, mutations of which, cause the human neurological disorder PHARC. Despite the biomedical significance of this lipase, detailed mechanistic studies and the specific contribution of ABHD12 to innate processes like phagocytosis remain poorly understood. Here, by immunohistochemical and immunofluorescence approaches, using the murine model of PHARC, we show, that upon an inflammatory stimulus, activated microglial cells in the cerebellum of mice deficient in ABHD12 have an amoeboid morphology, increased soma size, and display heightened phagocytosis activity. We also report that upon an inflammatory stimulus, cerebellar levels of ABHD12 increase to possibly metabolize the heightened oxidized PS levels, temper phagocytosis and in turn control neuroinflammation during oxidative stress. Next, to complement these findings, using biochemical approaches in cultured microglial cells, we show that the pharmacological inhibition and/or genetic deletion of ABHD12 results in increased phagocytic uptake in a fluorescent bead uptake assay. Together, our studies provide compelling evidence that ABHD12 plays an important role in regulating phagocytosis in cerebellar microglial cells, and provides a possible explanation, as to why human PHARC subjects display neuroinflammation and atrophy in the cerebellum.

Bacterial phagocytosis and reactive oxygen species production by camel neutrophils and monocytes are influenced by the type of anticoagulation agent

Background and Aim: Anticoagulants with different modes of action are used in the collection of camel blood samples. In the innate immune response, camel neutrophils and monocytes can play several roles during infection and inflammation. For anticoagulants ethylenediaminetetraacetic acid (EDTA) and heparin, research has described their effects on different parameters of the immune system. However, to date, no research has examined the effects of anticoagulants on the functional activity of camel phagocytes. Therefore, this study analyzed the influence of K3EDTA and lithium heparin on the antimicrobial activity of camel neutrophils and monocytes. Materials and Methods: Camel leukocytes were separated from blood collected in EDTA or heparin tubes, and their phagocytosis and reactive oxygen species (ROS) production activity were analyzed by flow cytometry after stimulation with Staphylococcus aureus or Escherichia coli bacteria. Results: In comparison to the cells collected from the EDTA blood, the camel neutrophils and monocytes separated from the heparin blood showed higher phagocytosis activity of S. aureus and E. coli. In addition, the neutrophils and monocytes produced significantly more ROS when the blood was collected in the heparin tubes. Conclusion: The antimicrobial functions of camel neutrophils and monocytes are significantly affected by the type of anticoagulation agent. Therefore, using heparin rather than EDTA as an anticoagulant is recommended when performing the functional analysis of phagocytosis and ROS production of camel phagocytes.

Immunotropic effects of nitrogenous metabolites in healthy humans

Background. We have previously shown that nitrogenous metabolites have immunomodulatory effects, both in healthy rats and in humans exposed to pathogenic influences. The purpose of this study is their immunotropic activity in clinically healthy people. Materials and Methods. The object of observation were 27 men (aged 24-63 ys) and 14 women (33-62 ys). The plasma levels of the nitrogenous metabolites and parameters of immunity twice with an interval of 5 days was performed. Results. Judging by the multiple correlation coefficient uric acid exhibits maximal immunotropic activity (R=0,665), followed by creatinine (R=0,596) and urea (R=0,541), and closes the constellation of metabolites bilirubin, with the activity of conjugated bilirubin predominating over that of unconjugated (0,539 vs 0,484). Nitrogenous metabolites together upregulate the level in the blood of B-lymphocytes, CIC, IgG, IL-1, eosinophils and monocytes, as well as most parameters of phagocytosis by neutrophils Staph. aureus and E. coli. Instead, they downregulate the phagocytosis activity of Staph. aureus, the relative content of rod-shaped neutrophils, lymphocytes in general and NK-, T active and 0-Lymphocytes in particular. Downregulation of 0-Lymphocytes reflects upregulation of receptor expression, apparently CD22. Conclusion. Nitrogenous metabolites exhibit immunotropic activity in both healthy rats and humans, as well as in patients.

Immunostimulant Activity of Gracilaria sp. and Padina sp. on Immune System of Vannamei Shrimp (Litopenaeus vannamei) Against Vibrio harveyi

The pathogenic bacterial infection is one of the problems in the cultivation of vannamei shrimp (Litopenaeus vannamei), causing a high mortality rate of cultured shrimp. The use of antibiotics or chemicals with inappropriate concentrations can harm the aquatic environment, cause resistance, and endanger consumer health because the residues from the chemicals used will periodically accumulate in the body of shrimp. One way to control and prevent shrimp disease is to increase the shrimp immune system by using immunostimulants from seaweed. This study aims to analyze the immunostimulant activity of seaweed extract (Gracilaria sp. and Padina sp.) against vannamei shrimp (L. vannamei) infected with Vibrio harveyi by observing the nonspecific immune system based on its hematological features, namely by counting the number of hemocytes and phagocytic activity. The research was conducted at the Hatchery Unit, Brackish Water Cultivation Development Center (BPAP) Situbondo, East Java. Seaweed sample Gracilaria sp. and Padina sp. purchased from seaweed farmers in Jepara, Central Java. The result of this study shows that supplementation of Gracilaria sp. and Padina sp. at a dose of 10 g/kg of feed can increase the total number of hemocytes and phagocytosis activity of L. vannamei shrimp. The best treatment is Gracilaria sp.

Primary culture macrophages from fish as potential experimental model in toxicity studies with multiwalled carbon nanotubes

Multiwalled carbon nanotube (MWCNT) has been broadly used in several sectors of society. This material when exposed to the environment might reach the aquatic animals and cause toxic effects. Here, it was evaluated the MWCNTs toxicity in melanomacrophages primary culture that was submitted to 1 µ gm L-1 MWCNTs for 24 hours. After exposition to MWCNT, 48 and 59% liver and spleen melanomacrophages were healthy, respectively. The control group presented 85% viability. Phagocytosis activity of melanomacrophages was observed by presence of black inclusions in cytoplasm. The findings indicate MWCNT was cytotoxic to melanomacrophages, where its release and effect into aquatic environment must be more studied. Finally, the melanomacrophages present large potential as experimental model for evaluation of carbon-based nanomaterial toxicity.