Suhartono Taat Putra
Departemen Patologi Anatomi Fakultas Kedokteran Universitas Airlangga

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KLONING GEN MELANOMA ANTIGEN 1 (MAGE-1) DARI JARINGAN TESTIS UNTUK MENDAPATKAN PLASMID REKOMBINAN MAGE-1 Mastutik, Gondo; I?tishom, Reny; Hardjowijoto, Sunaryo; Putra, Suhartono Taat
Majalah Kedokteran Bandung Vol 47, No 4 (2015)
Publisher : Faculty of Medicine, Universitas Padjadjaran

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Abstract

Gen Melanoma antigen-1 (Mage-1) diekspresikan oleh sel spermatogonia jaringan testis normal dan diekspresikan 60?80% oleh liver penderita karsinoma hepatoseluler (KH). Ekspresi Mage-1 merupakan penanda untuk diagnosis KH serta prediktor kanker lambung dan kolorektal. Isolasi messenger ribonucleid acid (mRNA) Mage-1 dari jaringan liver penderita KH sulit dilakukan sehingga dilakukan isolasi mRNA Mage-1 dari jaringan yang mengekspresikan Mage-1, yaitu jaringan testis normal. Penelitian ini merupakan penelitian eksploratif yang dilakukan di Lembaga Penyakit Tropis Universitas Airlangga, Agustus 2006?Agustus 2008. Tujuan untuk mengkloning seluruh area koding gen Mage-1 dari jaringan testis pada vektor dan mendapatkan plamid rekombinan Mage-1. Isolasi seluruh area koding gen Mage-1 dilakukan dengan teknik semi-nested polymerase chain reaction (PCR). Seluruh area koding gen Mage-1 diisolasi, kemudian dikloningkan ke plasmid pET101/D-TOPO dan ditransformasikan ke Escherichia coli (E. coli) Top10 untuk mendapatkan plasmid rekombinan Mage-1. Panjang pET101/D-TOPO adalah 5.753pb dan area koding gen penyandi Mage-1 927 bp sehingga total panjang plasmid rekombinan 6.680 bp (5.753+927). Hasil analisis restriksi dengan EcoRV menunjukkan pita 4.230 dan 2.450 (4.230+2.450 = 6.680). Analisis sekuens gen Mage-1 dari testis mempunyai homologi 100% dengan sekuens M77481 serta NM_004988, dan 99% dengan BC01755. Simpulan, berdasarkan hasil analisis restriksi dan sekuens maka diperoleh plasmid rekombinan pETGM/MAGE1-Testis yang mengandung seluruh area koding gen Mage-1 dan dapat digunakan untuk pengembangan kit diagnostik karsinoma.  [MKB. 2015;47(4):199?206]Kata kunci:  Jaringan testis, karsinoma hepatoseluler, kloning, melanoma antigen-1, pET101/D-TOPOCloning of Melanoma Antigen 1 (Mage-1) Gene from Testicular Tissue to Obtain the Recombinant Plasmid Mage-1Melanoma antigen-1 (Mage-1) is expressed by spermatogonia cells of normal testicular tissue and 60?80% is expressed by the liver of hepatocellular carcinoma (HC) patients. Mage-1 expression is a marker for diagnosing HC and predicting gastric and colorectal cancers. Isolation of messenger ribonucleid acid (mRNA) Mage-1 from the liver tissue of HC patients is difficult; therefore, Mage-1 mRNA isolates can be obtained from tissues that express Mage-1 such as normal testicular tissues . This is an explorative research that was conducted at the Institute of Tropical Diseases of Airlangga University during August 2006?August 2008. The aim was to clone the coding sequence of Mage-1 gene from testicular tissues into a vector and to get recombinant plasmid Mage-1. Isolation of the full-length Mage-1 was performed using semi-nested polymerase chain reaction (PCR) which was then cloned into plasmid pET101/D-TOPO and transformed into Escherichia coli (E. coli) Top10 to get recombinant plasmid Mage-1. The length of pET101/D-TOPO was 5,753 bp and Mage-1 was 927 bp. The length of recombinant plasmid was 6,680 bp (5,753+927). Restriction analysis using EcoRV showed 4,230 and 2,450 bp bands (4,230+2,450=6,680). Sequence analyses showed that Mage-1 was 100% homologous with M77481 and NM_004988, 99% homologous with BC01755. In conclusion, according to the results of the restriction and sequences analysis, the recombinant plasmid pETGM/MAGE1-Testis contains the full length coding region of Mage-1 and is useful for developing the hepatocellular carcinoma diagnostic kits. [MKB. 2015;47(4):199?206] DOI: 10.15395/mkb.v47n4.621
Increased Activity Of Mature Osteoblast from Rat Bone Marrow-Mesenchymal Stem Cells tn Osteogenic Medium Exposed to Melatonin Purnama, Yugi Hari Chandra; Mastutik, Gondo; Putra, Suhartono Taat
Folia Medica Indonesiana Vol 54, No 4 (2018): December
Publisher : Faculty of Medicine, Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (370.119 KB) | DOI: 10.20473/fmi.v54i4.10714

Abstract

Exposure to melatonin in the cultures of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) in osteogenic medium is able to induce mesenchymal stem cells and preosteoblasts into active osteoblasts via several transduction signals such as ERK 1/2. Previous studies used a single dose of 50 nM and a physiological dose of 20-200 pg/ml. The objective of the study was to obtain an optimal dose of melatonin that enhances osteoblast activity by increasing the expression of ERK1/2 and ALP levels in the culture of Rat Bone Marrow Mesenchymal Stem Cells (BM-MSCs) in osteogenic medium. This study was an in vitro experimental laboratory study using BM-MSCs from rat femoral bone grown on osteogenic medium without or with exposure to melatonin in doses of 0, 50, 100, 150 nM for 21 days. BM-MSCs were characterized by immunocytochemical techniques (CD45- and CD 105+) and ERK 1/2 expression was checked 24 hours after exposure to melatonin, while ALP levels were examined on day 21 using ELISA technique. ERK 1/2 expression on BM-MScs exposed to melatonin in doses 0, 50, 100, and 150 nM were respectively 0.087, 0.095, 0.081, and 0.079. Mean ERK 1/2 expression in various groups showed a decrease along with increasing doses of melatonin. Among the four treatment groups, the administration of melatonin in a dose of 50 nM resulted in highest mean ERK 1/2 expression. ALP levels in BM-MSCs exposed to melatonin doses of 0, 50, 100, and 150 nM were 0.128; 0.130; 0.117, and 0.111 ng/ml respectively. Data showed that decreasing mean ALP levels occurred along with the addition of melatonin dose. In conclusion, the administration of melatonin 50 nM is the optimal dose to increase the differentiation of cultured rat BM-MSCs into active osteoblasts.
SIREP SEBAGAI IMMUNOMODULATOR PADA LANSIA DENGAN GANGGUAN PEMENUHAN TIDUR Haryanto, Joni; Putra, Suhartono Taat
Jurnal Ners Vol 2, No 2 (2007): Oktober 2007 - Maret 2008
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (85.932 KB) | DOI: 10.20473/jn.v2i2.4967

Abstract

Introduction: Sirep is one of the nursing intervention to solve sleep problems. Moreover, sirep as immunomodulator for cortisol level, IFN-?Ra and IL-10 is still unknown.       Sleep disorder can make decrease of immunity, concentration disability, decrease of coordination, alteration of personality, etc. Decrease of immunity can caused by increase or imbalance of stress hormone (cortisol). Increase of cortisol in along time can suppresed T lymphocytes. Objective of this study was to analyze the effect of sleep therapy (sirep) as immunomodulator in elderly with sleep disorder Method: This study used Quasy-Experimental Pre-Post Test Control Group Non Randomised design. Result: Result showed that Sleep therapy (sirep) can changed IFNgRa level in serum elderly with Wilcoxon Sign Rank Test had p<0.01 and Mann Whitney U-Test had  p<0.01. Sleep therapy (sirep) can be change IL-10 level serum of elderly could be concluded that there was significant difference with Wilcoxon Sign Rank Test had p<0.01 and Mann Whitney U-Test not significant difference had  p>0.01. Discussion: Sleep therapy of elderly can released cortisol level serum changing, so sleep therapy as immunomodulator in elderly with sleep dissorder was verified. Discussion: Sleep therapy must applied on elderly with sleep disorder, because sleep therapy become immunomodulator on elderly with sleep disorder was verified.
Incidence of HPV Infection in Oral Squamous Cell Carcinoma and Its Association with the Presence of p53 & c-myc Mutation : A Case Control Study in Muwardi Hospital Surakarta Prayitno, Adi; Rukmo, Mandojo; Mudigdo, Ambar; Dalono, Dalono; Asnar, Elyana; Rahayu, Retno Puji; Purwanto, Joko Agus; Puspaningsih, Ni Nyoman Tri; Kuntoro, Kuntoro; Harjanto, Harjanto; Putra, Suhartono Taat
Journal of Dentistry Indonesia Vol 17, No 2 (2010): August
Publisher : Faculty of Dentistry, University of Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (487.302 KB) | DOI: 10.14693/jdi.v17i2.44

Abstract

Introduction: Annual incidence rates for oral and pharyngeal cancer are estimated at 25 cases per 100,000 in developing countries. Human papilloma virus (HPV) was implicated in pathogenesis of Cancer. The mutations of p53 and c-myc are found 50% in cancer. Objective: Aims of this research were to know the incidence of OSSC patient which realized HPV infection without p53 and c-myc gene mutation. Materials and Methods: Tissue biopsy frozen sections were taken from BOSC (Benign Oral Squamous Cell) and OSCC (Oral Squamous Cell Carcinoma) patients collected from Oral and Dental Departement of dr Muwardi Distric Hospital in Surakarta from January 2007 to January 2008. To amplify L1-HPV gene for fixed the HPV stressor. To amplify p53 and c-myc genes, continued with SSCP (Single Strand Conformational Polymorphisme) analysis and followed with measurement using densitometer, to see mutation existence. The collected data were analyzed with Chi Square. Results: BOSC patient identified 23% with HPV infections and OSCC patient identified 73% with HPV infections. Hundred percent BOSC patient with HPV infection without mutation in p53 gene and c-myc gene, 81% OSCC patient with HPV infection without mutation in p53 gene and 91% OSCC patient with HPV infection without mutation in c-myc gene. Chi  square analysis showed significant difference between BOSC and OSCC patients with HPV infection without mutation in p53 and c-myc gene. Conclusion: HPV is a factor for pathogenesis of OSCC.DOI: 10.14693/jdi.v17i2.44
The Role of Heat Shock Proteins in Pathogenesis of Oral Squamous Cell Carcinoma Prayitno, Adi; Rukmo, Mandojo; Mudigdo, Ambar; Dalono, J. B.; Asnar, Elyana; Rahayu, Retno Puji; Purwanto, Joko Agus; Puspaningsih, Ni Nyoman Tri; Kuntoro, Kuntoro; Harjanto, Harjanto J. M.; Putra, Suhartono Taat
Journal of Dentistry Indonesia Vol 16, No 2 (2009): August
Publisher : Faculty of Dentistry, University of Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (2754.761 KB) | DOI: 10.14693/jdi.v16i2.91

Abstract

Cell in the distress situation, denaturation of proteins may occur, and may also respond by expressing stress proteins. However, such homeostasis effort does not always succeed and even may lead to disease, including cancer. In distress situation also ensue much protein misfolding. Objective: This research were to explain the role of heat shock protein 40 (Hsp40) and Hsp70 in pathogenesis of occurred oral squamous cell carcinoma (OSCC) patient which realized human papilloma virus (HPV) infection. Material and Method: Tissue biopsy frozen section were taken from BOSC and OSCC patients was cut into three part. Parrafin blocks were made from cutting I, which was subsequently stains with HE to ascertain the type of neoplasm. Cutting II was subjected to DNA isolation. The DNA isolation results were subjected to PCR to amplify L1-HPV gene for fixed the HPV stressoor. Protein isolation was treated from Cutting III, folloewd with Blottdot test by using antibody monoclonal anti Hsp40 and Hsp70 and continued with measurement using densitometer to find the concentration of Hsp40 and Hsp70. The collected data were analyzed with F Test (Manova) and discriminant analysis. Result: This experiment showed the differences in concentration of Hsp40 (p&lt;=0,070) and Hsp70 (p&lt;=0,006) between beningn oral squamous cell (BOSC) and OSCC patients which realized HPV infection. Conclusion: This experiment proved that OSCC patients which realized HPV infection indicated an up regulated of Hsp70 concentration, so that there was occurs misfolding of the proteins cell. The misfolding was ensue obstacle of apoptosis and to raise cell proliferation which to storm carcinogenesis. An up regulated of Hsp40 was role as co-chaperone.DOI: 10.14693/jdi.v16i2.91 
Prevotella intermedia and Porphyromonas gingivalis in dental caries with periapical granuloma Cilmiaty, Risya; Prasetyo, Afiono Agung; Zaini, Khilyat Ulin Nur; Rukmo, Mandojo; Putra, Suhartono Taat; Asmara, Widya
Dental Journal (Majalah Kedokteran Gigi) Vol 46, No 4 (2013): (December 2013)
Publisher : Faculty of Dental Medicine, Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (492.71 KB) | DOI: 10.20473/j.djmkg.v46.i4.p213-217

Abstract

Background: Dental caries with necrotic pulp is a multifactorial disease that attacks enamel involving tooth pulp. The anaerobic bacteria infection in the pulp chamber could induce the formation of periapical granuloma. However, the presence of the most frequently anaerobic bacteria identified in apical periodontitis, Porphyromonas gingivalis and Prevotella intermedia, in periapical granuloma have not been confirmed. Purpose: The aims of study were to determine the presence of Porphyromonas gingivalis and Prevotella intermedia in dental caries with necrotic pulp and to determine its relation to periapical granuloma. Methods: Thirty-six patients of dental caries with necrotic pulp in Dr. Moewardi General Hospital in Surakarta, Indonesia were involved and classified into two groups, the group of patients with periapical granuloma and the group of patients without periapical granuloma. The caries tooth was extracted, and the chronic periapical tissue was swabbed and cultured on blood agar medium in anaerobic condition. The bacterial DNA was extracted from the positive cultures and subjected for Polymerase Chain Reaction (PCR). Results: Periapical granuloma was more likely found in women (OR 5.5, 95% CI=1.277-23.693; RR 2.5, 95% CI= 1.025-6.100). Black colonies bacteria were associated with periapical granuloma (OR 2.2, 95% CI=0.517-9.594; RR 1.5, 95% CI=0.655-3.623). Porphyromonas gingivalis and Prevotella intermedia were detected in group with or without periapical granuloma, however, only Prevotella intermedia was associated with periapical granuloma (OR 1.6, 95% CI=0.418-5.903; RR 1.3, 95% CI=0.653-2.393). Conclusion: The presence of Porphyromonas gingivalis and Prevotella intermedia in periapical granuloma were confirmed, however, only Prevotella intermedia were associated with periapical granuloma.Latar belakang: Karies gigi dengan pulpa nekrosis adalah penyakit multifaktorial yang menyerang enamel hingga ruang pulpa gigi. Infeksi bakteri anaerob di pulpa nekrosis dapat menginduksi pembentukan granuloma periapikal. Namun, keberadaan bakteri anaerob yang paling banyak ditemukan di periodontitis apikal, Porphyromonas gingivalis dan Prevotella intermedia, di granuloma periapikal masih perlu diteliti . Tujuan: Penelitian ini bertujuan meneliti keberadaan bakteri Porphyromonas gingivalis dan Prevotella intermedia, di karies gigi dengan pulpa nekrosis dan menganalisis kaitannya dengan granuloma periapikal. Metode: Tiga puluh enam pasien karies gigi dengan pulpa nekrosis di Rumah Sakit Umum Dr. Moewardi di Surakarta Indonesia dilibatkan dan diklasifikasikan dalam dua kelompok yaitu kelompok pasien dengan granuloma periapikal dan kelompok pasien tanpa granuloma periapikal. Gigi karies diekstraksi dan jaringan periapikal kronis diusap dan dikultur di media agar darah dalam kondisi anaerob. DNA bakteri diekstrak dari kultur yang positif dan dilakukan pemeriksaan dengan Polymerase Chain Reaction (PCR). Hasil: Granuloma periapikal lebih banyak ditemukan pada wanita (OR 5,5, 95% CI= 1,277-23,693; RR 2,5, 95% CI= 1,025-6,100). Koloni bakteri berwarna hitam diasosiasikan dengan granuloma periapikal (OR 2,2, 95% CI= 0,517-9,594; RR 1,5, 95% CI= 0,655-3,623). Porphyromonas gingivalis dan Prevotella intermedia terdeteksi di jaringan karies dental dengan atau tanpa granuloma periapikal, namun hanya Prevotella intermedia yang diasosiasikan dengan granuloma periapikal (OR 1,6, 95% CI= 0,418-5,903; RR 1,3, 95% CI= 0,653-2,393). Simpulan: Porphyromonas gingivalis dan Prevotella intermedia ditemukan di granuloma periapikal, namun hanya Prevotella intermedia yang terkait langsung dengan granuloma periapikal.
THE ELEVATION OF OSTEOBLAST ACTIVITY IN RAT BONE MARROW MESENCHYMAL STEM CELLS IN OSTEOGENIC MEDIUM EXPOSED WITH MELATONIN IN PHYSIOLOGICAL DOSES Yuliyanasari, Nurma; Mastutik, Gondo; Putra, Suhartono Taat
Folia Medica Indonesiana Vol 53, No 1 (2017): JANUARY - MARCH 2017
Publisher : Faculty of Medicine, Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (629.275 KB) | DOI: 10.20473/fmi.v53i1.5489

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The objective of this study was to analyze the elevation of osteoblast activity in bone marrow mesenchymal stem cells (BM-MSCs) in osteogenic medium by physiological doses of melatonin administration by measuring alkaline phosphatase (ALP) and osteocalcin level.This studyused BM-MSCs from Rattusnorvegiccus femur bone. Rat BM-MSCs were cultured in a-Mem medium, differentiated in osteogenic medium, and administrated melatonin up to 21 days. This study was divided into 4 groups, K0 (control group), K1 (administrated of 25 nM melatonin), K2 (administrated of 50 nM melatonin), and K3 (administrated of 100 nM melatonin). Rat BM-MSCs were characterized CD 45- and CD 105+ marker using imunocytochemistry analysis and stained with Alizarin red after 15 days treatment. ALP and osteocalcin level were measured using ELISA Kit in days 21st.There weren’t differences ofALP level beetwen groups and there are differences ofosteocalcin level between control groups (K0) withK1, K2, dan K3, and beetwen K1 and K2. The conclusion of this study was that there were an elevation of osteoblast activity in rat BM-MSCs in osteogenic medium by physiological doses of melatonin administration characterized by the elevation of osteocalcin level.
THE IMPROVEMENT OF FAMILY COPING IN TAKING CARE OF PATIENT MENTAL DISORDER WITH SPIRITUAL THERAPY; DIRECTION, OBEDIENCE AND ACCEPTANCE (DOA) Yusuf, Ah.; Putra, Suhartono Taat; Probowati, Yusti
Jurnal Ners Vol 7, No 2 (2012): Oktober 2012
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (230.338 KB) | DOI: 10.20473/jn.v7i2.4021

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Introduction: Mental disorder remains a stigma in society, even until now. A family who have a member with mental disorder, will experience continues objective and subjective burden, experience serious stress for a lifetime, which may cause ineffective coping.Method: Design used in this study was experimental (pre post test control group design). The population was every family of patient with mental disorder in Menur Mental Hospital along the year of 2010, has been taking care there twice, in minimum, lived in Surabaya. The samples were chosen by allocation simple random. Samples were 13 persons in each treatment and control group. The intervention was given in 60?120 minute in 8 times meeting with average interval about 1 week. Data analysis was done using paired t-test and independent t-test.Result: Results in this study showed that there was signi?cant change in total of family coping (p = 0.040), maintaining family integration, cooperation and an optimistic de?nition of the stuation (p = 0.009), maintaining social support, self esteem, and psychological stability (p = 0.230), understanding the medical situations through communication with other parents and concultation with medical staff (p = 0.025).Discussion: The provision of family therapy with spiritual approach (DOA) can increase family coping in taking care of patient with mental disorder.
TRADITIONAL ACTIONS: SIREP INFLUENCE CORTISOL, IFN- AND IL-10 IN ELDERLY WITH SLEEP DISORDERS Haryanto, Joni; Putra, Suhartono Taat
Jurnal Ners Vol 11, No 2 (2016): Oktober 2016
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (286.436 KB) | DOI: 10.20473/jn.V11I22016.277-282

Abstract

Introduction: Sirep, is one of the traditional intervention of people in East Java Indonesia, especially Tengger tribe using spells to meet the needs of human sleep. Elderly in Indonesia is someone who has more than 60 years of age. Elderly in general often experience sleep disturbances fulfillment, But the effects of nursing Sirep as an immunomodulator to the levels of cortisol, IFN-?R?  and IL-10 is still unknown in nursing perspective. Method: The population of this study were 70 elderly with the number of respondents were 43 elderly. They were divided into two groups for 23 elderly with 6-month sirep intervention (mean age, 69.6 ± 5.2 years) as an experiment group  and  20 elderly as control group (65.8 ± 5.4 years). The need of sleep is implemented to Evaluate the sample of immune modulation, cortisol levels, IFN-?R?  and IL10. The molecular biology of the samples was determined using  ELISA analysis. The statistical analysis of the data used Determine ANOVA and t-test. Results: After the 6 -month intervention, there re is significant changes in serum cortisol levels (p = 0.0001) and a different test with a control intervention was also significant (p = 0.00 1 3). Significant changes in serumlevels of IFN-?R?  is (p = 0.0003) and a different test with a control intervention is also significant (p = 0.016 4), while the change in the levels of IL-10 in serum is significant (p = 0.0003) and test different intervention with no significant control (p = 0.1143). Discussion: This study supports the hypothesis that sirep can improve immune modulator in elderly. However, immune modulator showed a slight improvement in the nursing intervention of sirep group, suggesting that a longer or more frequent sessions of intervention might have an effect.
PSYCHONEUROIMMUNOLOGY APPROACH TO IMPROVE RECOVERY MOTIVATION, DECREASE CORTISOL AND BLOOD GLUCOSE OF DM TYPE 2 PATIENTS WITH DHIKR THERAPY Pahlevi, Rifka; Putra, Suhartono Taat; Sriyono, Sriyono
Jurnal Ners Vol 12, No 1 (2017): April 2017
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (212.955 KB) | DOI: 10.20473/jn.v12i1.2315

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Introduction: Blood glucose levels are controlled when the management of diabetes success. Positive perception of the strength of the spiritual aspect will improve the motivation of patients with type 2 diabetes to control it. The purpose of this study was to determine the effect of dhikr based on psychoneuroimmunology (PNI) on blood glucose levels of patients with type 2 diabetes.Methods: This study used quasi-experiment with pre-test and post-test control group design. Samples were taken from the population of patients with type 2 diabetes who were hospitalized in the Internal Medicine Rumkital Dr. Ramelan Surabaya with purposive sampling techniques. Data taken include the general characteristics of respondents, cures motivation, cortisol levels and fasting blood glucose levels. Collecting data using questionnaires and laboratory test, then analyzed using paired t-test and independent t-test, with ? value <0.05.Results: Statistical test showed that the motivation to recover increased (p = 0.001), cortisol levels fall (p = 0.058) and a drop in blood glucose levels (p = 0.028) after administration of dhikr therapy in patients with diabetes mellitus type 2. There was a significant difference in increased of recovery motivation between patient conduct zikr therapy and patient cared (p = 0,000).Conclusion: Dhikr therapy increases the motivation of patients with type 2 diabetes by strengthening awareness and spirituality belief in Allah make positive stress perception. Positive stress perception will affect the stress response and improved regulation of blood glucose through the HPA axis to suppress the secretion of CRH, ACTH, and cortisol.