Chicken Immune Cell Assay to Model Adaptive Immune Responses In Vitro

Knowledge about the modes of action of immunomodulating compounds such as pathogens, drugs, or feed additives, e.g., probiotics, gained through controlled but animal-related in vitro systems using primary cultured peripheral blood mononuclear cells (PBMCs) will allow the development of targeted nutrition strategies. Moreover, it could contribute to the prevention of infectious diseases and the usage of antimicrobials, and further promote the health of the animals. However, to our knowledge, a protocol for the isolation of PBMCs with reduced thrombocyte count from chicken blood and subsequent cell culture over several days to assess the effects of immunomodulating compounds is not available. Therefore, we established an optimized protocol for blood sampling and immune cell isolation, culture, and phenotyping for chicken PBMCs. For blood sampling commercial Na–citrate tubes revealed the highest count of vital cells compared to commercial Li–heparin (p < 0.01) and K3EDTA (p < 0.05) tubes. Using combined dextran and ficoll density gradient separation, the thrombocyte count was significantly reduced (p < 0.01) compared to slow-speed centrifugation with subsequent ficoll. For cell culture, the supplementation of RPMI-1640 medium with 10% chicken serum resulted in the lowest relative cell count of thrombocytes compared to fetal calf serum (FCS) (p < 0.05). To validate the ability of the cell culture system to respond to stimuli, concanavalin A (conA) was used as a positive control. The optimized protocol allows the isolation and cultivation of vital PBMCs with reduced thrombocyte count from chicken blood for subsequent investigation of the modes of action of immunomodulating compounds.

The Demography, Clinical Characteristics, and White Blood Analysis of Leprosy Reactions in Multibacillary Leprosy: A Retrospective Study

Background: Leprosy is a neglected tropical disease caused by chronic granulomatous infection of Mycobacterium leprae. Indonesia ranks third in new case findings, with 84% of the case being multibacillary (MB) leprosy. MB leprosy cases have a higher risk of leprosy reactions and physical disabilities that decrease quality of life. Purpose: To determine the demographic, clinical characteristics, and white blood analysis of newly diagnosed MB leprosy patients, especially concerning leprosy reactions. Methods: This is a descriptive retrospective study with a cross-sectional design that describe the following data: domicile, gender, age, treatment status, disabilities, body mass index (BMI); bacterial index (BI), morphological index (MI), white blood cell (WBC) and differential counts, and thrombocyte count. Result: This study included 176 adult MB cases, predominantly male aged 20–39 years old with average BMI, lived in Surabaya with negative history of multi-drug therapy, disability, BI, nor MI. The grade 2 disability (G2D) percentage in this study setting than in Indonesia (10.7% vs. 6.43%). The WBCs, especially neutrophil count, was higher in T2R group. Monocyte and lymphocyte counts were relatively similar. There was an increase in thrombocyte count in leprosy reaction groups. Conclusion: MB leprosy in the endemic area, which is more commonly found in productive-aged male, displayed higher G2D than global Indonesia population. Thus denotes the importance of active case findings. The difference in blood analysis characteristics between MB leprosy with and without reactions may serve as the foundation for future study.

Coagulation parameters in the newborn and infant – the Copenhagen Baby Heart and COMPARE studies

Objectives The coagulation system is not fully developed at birth and matures during the first months of infancy, complicating clinical decision making within hemostasis. This study evaluates coagulation parameters at birth and two months after birth, and tests whether cord blood can be used as a proxy for neonatal venous blood measurements. Methods The Copenhagen Baby Heart Study (CBHS) and the COMPARE study comprise 13,237 cord blood samples and 444 parallel neonatal venous blood samples, with a two month follow-up in 362 children. Results Because coagulation parameters differed according to gestational age (GA), all analyses were stratified by GA. For neonatal venous blood, reference intervals for activated partial thromboplastin time (APTT) and prothrombin time (PT) were 28–43 s and 33–61% for GA 37–39 and 24–38 s and 30–65% for GA 40–42. Reference intervals for international normalized ratio (INR) and thrombocyte count were 1.1–1.7 and 194–409 × 109/L for GA 37–39 and 1.2–1.8 and 188–433 × 109/L for GA 40–42. Correlation coefficients between umbilical cord and neonatal venous blood for APTT, PT, INR, and thrombocyte count were 0.68, 0.72, 0.69, and 0.77 respectively, and the distributions of the parameters did not differ between the two types of blood (all p-values>0.05). Conclusions This study describes new GA dependent reference intervals for common coagulation parameters in newborns and suggests that cord blood may serve as a proxy for neonatal venous blood for these traits. Such data will likely improve clinical decision making within hemostasis among newborn and infant children.

Ekstrak Rumput Laut Sargassum.sp Mencegah Trombositopenia Gestational Pada Tikus Selama Kebuntingan

The potential of seaweed as a functional food ingredient has not been explored. Several studies, the compotition of seaweed can prevent stress in animals such as sedative mechanisms. The sedative effect was decreased of blood pressure. Pregnancy can be a stressor in animals and cause anemia physiologically. This study aims to determine the effect of seaweed extract on the profile hemoglobin and erythrocytes of pregnant mice. In this study used 10 pregnant rats divided into 2 groups, group 1 (control) and group II (treatment with 450 mg/BW extract Sargassum sp). Calculation of erythrocytes, hemoglobin, and thrombocyte measurements were carried out in day 0, 7, and 14. The results showed that there was an increased thrombocyte count in the treatment group even though in both groups there was a decreased the number of erythrocytes and hemoglobin. The results of statistical analysis showed a significant difference (p <0.05) of thrombocyte counts between the control and treatment groups. Based on the results of the study it was found that administration of Sargassum sp. during pregnancy can prevent thrombocytopenia in pregnant rat. Keywords: Sargassum sp; Pregnant rat; Thrombocytopenia; Erythrocytes; Hemoglobin Abstrak Potensi rumput laut sebagai bahan pangan fungsional belum banyak digali. Beberapa penelitian telah menyebutkan kandungan rumput laut dapat mencegah stress pada hewan seperti mekanisme sedativa. Efek sedativa yang sering muncul adalah penurunan tekanan darah. Kebuntingan dapat merupakan stressor pada hewan dan secara fisiologi akan menyebabkan anemia. Penelitian ini bertujuan untuk mengetahui efek pemberian ekstrak rumput laut pada gambaran hematologi tikus bunting. Penelitian ini menggunakan 10 ekor tikus bunting yang dibagi menjadi 2 kelompok yaitu kelompok 1 (kontrol) dan kelompok II (perlakuan ektrak rumput laut Sargassum sp 450 mg/kg BB). Pengambilan sampel dilakukan pada hari ke 0 (sebelum kebuntingan), hari ke 7 dan hari ke 14 (akhir kebuntingan). Pengukuran hematologi sampel meliputi perhitungan eritrosit, pengukuran hemoglobin serta trombosit. Hasil penelitian menunjukkan bahwa terjadi peningkatan jumlah trombosit pada kelompok perlakuan meskipun pada kedua kelompok terjadi penurunan jumlah eritrosit dan hemoglobin. Hasil analisis statistik ada perbedaan yang signifikan (P<0,05) jumlah trombosit antara kelompok kontrol dengan perlakuan. Berdasarkan hasil penelitian didapatkan bahwa pemberian ekstrak Sargassum sp selama kebuntingan dapat mencegah terjadinya trombositopenia pada tikus bunting. Kata kunci: Sargassum sp; Tikus bunting; Trombositopenia; Eritrosit; Hemoglobin

The Prevalence of Reactive Thrombocytosis and Thromboembolic Events in Arab Population with Iron Deficiency Anemia (a retrospective study)

INTRODUCTION Iron deficiency is the most prevalent nutritional deficiency worldwide. Iron deficiency anemia (IDA) is the most common type of anemia, its prevalence is 1 out of 5 of the population. IDA is a well-known cause of reactive thrombocytosis which is mostly asymptomatic. Only few observational studies and case reports have described thromboembolic events in the context of this reactive thrombocytosis in the absence of other hypercoagulable states OBJECTIVES To assess the frequency of thromboembolic events in Arab patients with reactive thrombocytosis secondary to iron deficiency anemia (IDA). METHODS We retrospectively reviewed thromboembolic events in iron-deficient patients with reactive thrombocytosis. Our study sample included female patients who received iron replacement for IDA between April 2018 and March 2020 at Hamad Medical Corporation, Doha, Qatar. We excluded pregnant, non-Arab patients and patients under 18 or over 65 years of age. Reactive thrombocytosis was defined as thrombocyte count of more than or equal to 450 x 109 /L in the presence of iron deficiency anemia and ferritin level less than 30µg/l. RESULTS Out of 1567 patients (mean age =50 +/- 8.66 years) with the diagnosis of IDA, 292 (18.63%) had thrombocytosis. They had a mean platelet count = 534 +/- 121 x 109 /L). None of them had any symptom or sign of thromboembolic events. Discussion Thrombocytosis can be categorized into primary causes such as primary bone marrow disorders and myeloproliferative neoplasms, and secondary causes including infection, inflammation or drug-induced. Iron deficiency anemia leads to reactive thrombocytosis in a mechanism that is yet to be fully understood. The clinical impact of increased platelet counts is not well recognized in literature, and it has not been studied in the Arab population. Although reactive thrombocytosis has been generally considered benign, few case reports described thromboembolic events in patients with IDA reactive thrombocytosis. Few cases in the literature described thromboembolic events in reactive thrombocytosis with IDA. H. Z. Batur Caglayan et al published a case report of a 41-year-old Turkish female patient who presented with transient ischemic attack (TIA) due to intraluminal carotid artery thrombus, which was attributed to IDA-associated thrombocytosis. Another case series by P T Akins et al described three women with severe IDA and thrombocytosis secondary to menorrhagia who developed carotid artery thrombi. The mechanism by which low iron can affect thrombocyte count is still unknown. One study in mice and humans demonstrated that iron deficiency caused reduced megakaryocyte proliferation but increased ploidy independent of thrombopoietin. However, another study failed to identify the exact mechanism by which iron deficiency leads to increased platelet count. CONCLUSION Our study did not find any thromboembolic incident in a large number of patients with reactive thrombocytosis secondary to IDA diagnosed over two years in our Arab population. Disclosures No relevant conflicts of interest to declare.

Thrombocyte Count in Male and Female Adult of Dengue Hemorrhagic Fever Patients

The trend of dengue virus infection in Indonesia has changed from children to older groups. Some studies suggested that different gender might affect the different progression of dengue infection, but its mechanism was unclear. This study analyzed the difference by evaluating the daily thrombocyte count pattern. An analytic observational study with a retrospective design was conducted using the secondary data collected from medical records in Dr. Soetomo General Hospital Surabaya during 2017-2018. The samples were adult patients in the age range of 18-55 years old with DHF but without comorbidity or coinfection. The total number of samples was 40 patients. The average of thrombocyte count in both male and female patients decreased since the 3rd day of illness and reached the lowest level on the 6th day of illness then increased on the 7th day but did not reach the normal range. Although the majority of thrombocyte count in females was lower than males, there was no significant difference in thrombocyte count pattern between them. Keyword: Adult, DHF, Thrombocyte, GenderCorrespondence : [email protected]

Exploration of prognostic factors for critical COVID-19 patients：a nomogram analysis

To discuss influencing factors on critical COVID-19 patient’s prognosis, construct a basic model and predict their mortality risks. Retrospectively analyzed the general condition and respective laboratory biomarkers of critical patients with duration≥24 h from Feb. 10th, 2020 to Mar. 30th, 2020 to separate them into a survival group and death group based on their clinical features. Multiple logistic regression analysis was performed to assess risk factors for critical COVID-19 patient’s and a nomogram was constructed based on screened risk factors. A receiver operating curve (ROC) was created to evaluate the accuracy of the nomogram. Multi-factor Logistic recovery analysis results show: Age, Peripheral blood leucocyte count,Lymphocyte percentage, Thrombocyte count and Hyper C-reactive protein are single danger factors of critical COVID-19 patient’s mortality risk (p＜0.05). ROC curve indicates Nomogram predictive model AUC is 0.958 (95%CI: 0.923-0.993), which has high predictive value. Findings from this study suggest advanced age, high peripheral blood leucocyte count, high hyper C-reactive protein, low lymphocyte percentage and low thrombocyte count are risk factors of critical COVID-19 patient’s death.The Nomogram model is helpful for timely intervention to reduce the incidence of critical COVID-19 patients.

Determining Postoperative Thrombocyte Reduction Cut Off to Predict 30-Day Mortality in Patients Undergo Coronary Artery Bypass Graft Surgery

Background There are many factors associated with early mortality after CABG, including postoperative thrombocytopenia (Kertai, 2016). Many factors during CABG surgery, such as administration of heparin or cardio pulmonary bypass during surgery are related to thrombocyte count reduction (Hamid, Akhtar, Naqvi, & Ahsan, 2017; Arepally, 2017). However, it is possible for a post-CABG patient to suffer a significant thrombocyte reduction without reaching the thrombocytopenic state (thrombocyte count <150000/µL). Up to this time, there is still lack of study about association between thrombocyte reduction after surgery and 30-day mortality in patients undergo CABG. This study aim to determine cut off point for postoperative thrombocyte reduction as a predictor of 30-day mortality after CABG surgery. Method This is a retrospective cohort study using medical record of 263 adult patients who underwent CABG surgery in dr. Ciptomangunkusumo National Hospital on 2012-2015. Thrombocyte reduction was determined by substracting preoperative thrombocyte count from postoperative thrombocyte count. Receiver operating curve (ROC) analysis between percentage of thrombocyte reduction and 30-day mortality after surgery was done to obtain the sensitivity and specificity value of a particular degree of thrombocyte reduction. Cut off point was obtained from intersection between sensitivity and specificity value. Result Thirty-day mortality rate after CABG surgery in this study was 11.9%. Cut off point obtained from ROC analysis was 30% with area under the curve (AUC) 0.671. The sensitivity of this cut off point to predict early mortality after CABG surgery was 64.5%, while the specificity was 64.7% Conclusion Thrombocyte reduction more than or equal to 30% can be used as a predictor of 30-day mortality after CABG surgery. Figure Disclosures No relevant conflicts of interest to declare.

DESKRIPSI HEMATOLOGI KELELAWAR DI GUNUNG TANGKOKO BERDASARKAN PEMERIKSAAN DARAH LENGKAP

ABSTRACTThis study aimed to describe the hematological data of several bat species netted on Mount Tangkoko based on a complete blood examination. Purposive Random Sampling method was used for field sampling, Mistnet method was used for bats sampling and Screening Test method was used for hematological examination. Blood was examined using the Abx Micros 60 Hiruba Medical machine. The examined blood components were erythrocytes, leukocytes, haemoglobin, haematocrit, and platelets/thrombocytes. The examination results showed that each blood component had different amount compared to humans, some other mammals with relativity in size and some other birds. The highest leukocyte count was found in T. nigrescens which was 28.00 ± 19.04 × 103/mm3. The highest erythrocyte count was found in T. nigrescens which was 13.45 ± 1.52 × 106/mm3. The highest haemoglobin and haematocrit count were found in R. amplexicaudatus which were 19.70 ± 1.89 g/dl and 65.70 ± 5.65%. The highest platelet/thrombocyte count was found in T. nigrescens which was 975.2 ± 239.03 × 103/mm3.Keywords: Bats, Mount Tangkoko, Erythrocytes, Leukocytes, Haemoglobin, Haematocrit, Platelets ABSTRAKPenelitian ini bertujuan untuk mendeskripsikan data hematologi beberapa spesies kelelawar yang terjaring di Gunung Tangkoko berdasarkan pemeriksaan darah lengkap. Sampling lokasi menggunakan metode Purposive Random Sampling, penangkapan kelelawar menggunakan metode Mistnet dan pemeriksaan hematologi menggunakan metode Screening Test. Darah diperiksa menggunakan mesin Abx Micros 60 Hiruba Medical. Hasil menunjukan lima komponen darah yaitu eritrosit, leukosit, hemoglobin, hematokrit, dan platelet/trombosit masing-masing memiliki perbedaan dengan manusia dan beberapa spesies mamalia lain dengan ukuran yang relatif sama serta beberapa spesies burung sebagai pembanding. Nilai leukosit tertinggi terdapat pada spesies T. nigrescens yaitu 28,00 ± 19,04 × 103/mm3. Nilai eritrosit tertinggi terdapat pada spesies T. nigrescens yaitu 13,45 ± 1,52 × 106/mm3. Nilai hemoglobin dan hematokrit tertinggi terdapat pada spesies R. amplexicaudatus yaitu masing-masing 19,70 ± 1,89 g/dl dan 65,70 ± 5,65%. Nilai platelet/trombosit tertinggi terdapat pada spesies T. nigrescens yaitu 975,2 ± 239,03 × 103/mm3. Kata kunci : Kelelawar, Gunung Tangkoko, Leukosit, Eritrosit, Hemoglobin, Hematokrit, Leukosit