Difference between Microscopic and PCR Examination Result for Malaria Diagnosis and Treatment Evaluation in Sumba Barat Daya, Indonesia.

Microscopic examination is the backbone of malaria diagnosis and treatment evaluation in Indonesia. This test has limited ability to detect malaria at low parasite density. Inversely, nested polymerase chain reaction (PCR) can detect parasites at a density below the microscopic examination's detection limit. The objective of this study is to compare microscopic and PCR results when being used to identify malaria in suspected patients and patients who underwent dihydroartemisinin-piperaquine (DHP) therapy in the last 3-8 weeks with or without symptoms in Sumba Barat Daya, Nusa Tenggara Timur, Indonesia. Recruitment was conducted between April 2019 and February 2020. Blood samples were then taken for microscopic and PCR examinations. Participants (n = 409) were divided into three groups: suspected malaria (42.5%), post-DHP therapy with fever (4.9%), and post-DHP therapy without fever (52.6%). Microscopic examination found five cases of P. falciparum + P. vivax infection, while PCR found 346 cases. All microscopic examinations turned negative in the post-DHP-therapy group. Conversely, PCR result from the same group yielded 29 negative results. Overall, our study showed that microscopic examination and PCR generated different results in detecting Plasmodium species, especially in patients with mixed infection and in patients who recently underwent DHP therapy.

Proteomic networks associated with tumor-educated macrophage polarization and cytotoxicity potentiated by heat-killed tuberculosis.

Local administration of attenuated mycobacterium has been used as a cancer treatment adjuvant to re-boost patient immune responses with variable clinical outcomes. We aimed to clarify the impact of attenuated heat-killed tuberculosis (HKTB) on tumor-associated macrophages which play critical roles in shaping immunological regulation in the tumor microenvironment. Upon HKTB stimulation, both primary macrophages derived from the peripheral blood of healthy subjects and from lung cancer patients as well as THP1-derived classically activated macrophages (Ms) and tumor-educated macrophages (TEMs) were polarized into the proinflammatory phenotype, as characterized by increased expression cluster of differentiation 86. A quantitative proteomic analysis revealed that stimulated TEMs were unable to activate the toll-like receptor 2, signal transducer and activator of transcription 1, or nuclear factor-κB signaling. Instead, they showed distinct intercellular adhesion molecule 1 signaling, impaired cell adhesion, and mitochondrial dysfunction. These molecular mechanisms might contribute to lower cytotoxicity of HKTB-stimulated TEMs against A549 cells via the release of distinct inflammatory cytokines compared to HKTB-stimulated Ms. Our study provides an unbiased and systematic interpretation of cellular and molecular alterations of HKTB-reeducated macrophages which should help illuminate potential strategies of HKTB-stimulated macrophage-based combination therapy for cancer treatment.

The Role of Mannose-Binding Lectin Serum Level in Tubotympanic Chronic Suppurative Otitis Media.

BACKGROUND: Chronic suppurative otitis media (CSOM) is a common public health problem worldwide and a major cause of hearing impairment especially in developing countries. The role of Mannose-Binding Lectin (MBL), a component of innate immunity, in CSOM has not been studied. The aim of the study was to examine whether MBL deficiency was more frequently present in cases group of tubotympanic CSOM patients rather than healthy subjects. MATERIAL AND METHODS: This was an analytic observational study. Subjects were enrolled in the Otorhinolaryngology Clinic at Margono Soekarjo Hospital, Purwokerto, Indonesia. An independent t-test was used to compare the mean of MBL serum concentration between tubotympanic CSOM subjects and control. RESULTS: From 36 tubotympanic CSOM patients, there were 8 (22.22%) patients with MBL deficiency (MBL level < 100 ng/ml), while no deficiency was found in the control group. The mean of MBL level in cases group was 354.88 ng/ml, with the lowest level being 0.001 ng/ml and the highest level 690.24 ng/ml, while in the control group MBL level mean was 376.27 with the lowest level being 188.71 and the highest level 794.54 ng/ml. CONCLUSION: There was no significant difference of MBL serum level between tubotympanic CSOM and control group. However, the presence of subjects with MBL deficiency in the tubotympanic CSOM group might be considered as playing a role in the tubotympanic CSOM.

Isoniazid (INH) treatment of INH-resistant Mycobacterium tuberculosis inhibits infected macrophage to produce TNF-alpha.

Macrophages undergo apoptosis after infected by Mycobacterium tuber- culosis (M.tb), which is regulated by tumor necrosis factor alpha (TNF-alpha) and has a direct correlation with killing of intracellular bacilli. In order to clarify the role of isoniazid (INH) in the modulation of macrophages apoptosis and intracellular bacilli replication, we performed the following studies using an INH-resistant clinical M.tb isolate (INHres). Macrophages derived from peripheral blood were infected with INHres and treated or not treated with INH. Apoptosis was measured using an Ag-capture ELISA for histone and fragmented DNA. Production of TNF-alpha by INHres infected was assayed using ELISA and viability of intracellular M.tb was determined using bacterial culture of macrophage lysates on Lowenstein-Jensen (LJ) medium. INH pre-treatment to INHres reduced macrophages apoptosis, production of TNF-alpha and intracellular INHres viability. This study indicated that INH affected TNF-alpha release resulting in reduction of the extent macrophages apoptosis and of intracellular INH-resistant M.tb viability.

The effects of oral plain kefir supplementation on proinflammatory cytokine properties of the hyperglycemia Wistar rats induced by streptozotocin.

AIM: to validate the effect of plain kefir on immune responses of hyperglycemia wistar rats induced by Streptozotocin. METHODS: the randomized pretest - posttest control group study design was conducted in male hyperglycemia Wistar rats induced by streptozotocin (STZ). Rats were randomized into four groups: (1) STZ-induced group were given insulin treatment 0.76 UI/200 g bw, (2) STZ-induced group and treated with plain kefir 3.6 cc/200 g bw/day for 30 days, (3) STZ-induced group as control, (4) normal animal group as a negative control. Blood glucose was measured from whole blood that was taken 0.1 ml from retroorbitalis vein by microhematocrit on day 1 (pretest) and day 30 (post test) by enzymatic methods. Immune responses (cytokines IL1, IL6, IL10, TNF) were measured by ELISA. Data were analyzed by one way Anova, Mann Whitney test and Duncan with significant level of p<0.05. RESULTS: plain kefir supplementation 3.6 cc/day affect blood glucose, proinflamatory cytokines (IL1, IL6, TNF) and antiinflamatory cytokine (IL10). Statistical analysis showed decrease of glucose -111.00±44.23 ml (p<0.001) and proinflamatory cytokines IL1 about -18.62±23.59 and IL6 -3.21±7.57 mU/mL (p<0.001), respectively compared to the control groups. TNF decreased 1.65±4.62 mU/mL, but not significant (p>0.05), except for controls group. In addition, antiinflammatory (IL10) showed also increase about 15.11±2.16 (p<0.05), except for the control. CONCLUSION: plain kefir supplementation significantly decreased blood glucose, level of cytokines (IL1, IL6) and lowered TNF level. On the contrary, the level of IL10 is increased compare to control groups.

Relationship of angiotensin converting enzyme gene polymorphism and hypertension in Yogyakarta, Indonesia.

AIM: To evaluate the association between ACE gene polymorphism I/D and hypertension in Yogyakarta population. METHODS: This study is a cross-sectional. Sample was taken by random sampling method from hypertensive, prehypertensve and normotensive subjects; from that were obtained 125 subjects, 97 subjects and 108 subjects, consecutively. ACE gene polymorphism I/D was examined by PCR. Genotype was classified as II, ID, or DD based on positive or negative insertion/delation allele. RESULTS: This study shows significant differences of three groups (ages, body mass index (BMI), and family history of hypertension) and total cholesterol level in blood, which tends to have greater value in the hypertension group. Frequency of genotype II, ID, DD are 85 (68%), 39 (31.2%), 1 (0.8%) in hypertension, 66 (61.1%), 38 (35.2%), 4 (3.7%) in normo-tension and 56 (57.7%), 37 (38.1%), 4 (4.1%) in pre-hypertension subject, consecutively. Chi-square analysis shows statistically significant association between ID+DD vs. II genotype and hypertension. Multiple logistic regression analysis shows four variables that significantly influence to hypertension, namely ages, family history of hypertension, BMI, and ACE gene polymorphism. CONCLUSION: ACE ID+DD genotype has significant relationship with hypertension in Melati population, Sleman, Yogyakarta, Indonesia.

The role of CD4+ T cells in the induction of contact hypersensitivity to mercury in a murine model.

BACKGROUND: The exact immunopathogenesis of contact hypersensitivity (CH) to mercury remains unclear. OBJECTIVE: The aim of the present study was to assess the role of CD4+ T cells in mercury-induced CH in mice. METHODS: Splenic CD4+ T cells obtained from nonsensitized and mercury-sensitized Balb/c mice were adoptively transferred to groups I and II of the syngeneic recipients, respectively. All recipients were ear-challenged with mercury. The next experiments were to transfer nonsensitized CD4+ T cells to group A of the recipients, whereas mercury-sensitized CD4+ T cells were transferred to groups B and C. Groups A and B were ear-challenged with mercury, whereas group C was ear-challenged with chromium. The ear swelling 24 hours and 48 hours after challenge was used to assess CH. RESULTS: The results of the present study showed that mercury-sensitized but not nonsensitized CD4+ T cells could transfer CH in mice. Furthermore, mercury-sensitized CD4+ T cells could transfer the induction of CH only in the recipients that were challenged with mercury, but not those that were challenged with chromium. CONCLUSION: The results of the present study suggest that mercury-induced CH may be mediated by mercury-specific CD4+ T cells.

Interleukin-10 levels in Epstein-Barr virus-associated nasopharyngeal carcinoma.

The aim of this study was to determine the levels of interleukin-10 in patients with nasopharyngeal carcinoma. Both biopsies and sera were obtained from patients with nasopharyngeal carcinoma as well as Epstein-Barr virus-seronegative patients as a control. Nasopharyngeal carcinoma patients were classified using the World Health Organization pathological assessment and clinical staging of nasopharyngeal carcinoma. The numbers of interleukin-10 positive cells and the levels of serum interleukin-10 were assessed by immunohistochemical methods and enzyme-linked immunosorbent assay, respectively. The levels of serum interleukin-10 were determined by enzyme-linked immunosorbent assay. The results showed that the number of interleukin-10 positive cells and serum interleukin-10 levels were significantly increased in patients with nasopharyngeal carcinoma-World Health Organization type III and with clinical late stage (p<0.05), suggesting that interleukin-10 may have a crucial role in the progression of nasopharyngeal carcinoma.