Capillary Centrifugation Research Articles

Analysis of RhC Antigen Weak Expression Combined with Mimicking Autoanti-Ce and Homologous Anti-Jkb Causing Mismatch

To investigate the reasons for the weak expression of RHCE gene in a patient whose mimicking anti-Ce combined with anti-Jkb caused cross-matching non-combination. ABO, Rh, and Kidd blood group antigens were identified by test tube method and capillary centrifugation. Antibody screening and antibody specificity identification were performed using saline, polybrene and antiglobulin in tri-media association with multispectral cells. RHCE gene sequencing and haploid analysis were performed by multiplex PCR technique and RHCE protein modeling was performed using Swiss-Model. The serum of the patient contained anti-Ce mimicking autoantibodies along with anti-Jkb antibodies. c.48G>C, c.150C>T, c.178C>A, c.201A>G, c.203A>G, and c.307C>T mutations were detected in the RHCE triple-molecule sequencing. A 109 bp insertion sequence was found in intron 2, with fragment loss from intron 5-8. The Rh-group genotype was DCe/DCe , and phenotype was CCDee. Genotyping techniques can assist in deducing the molecular mechanisms of some weakly expressed RhC, c, E, and e in patients' sera to aid in the identification of difficult antibodies and thus ensure the safety of patients' blood transfusion.

Hematocrit estimates comparing centrifugation to a point-of-care method in beef cattle living at high altitude.

The hematocrit (Hct) or packed cell volume (PCV) reflects the blood volume occupied by red blood cells. The development of point-of-care (PoC) instruments can accelerate the ease of measuring Hct/PCV compared with traditional capillary centrifugation (TCC) methods. However, no studies have compared Hct/PCV levels in cattle at high elevation with other measurement methods. In this study, we aimed to compare methods to estimate Hcts/PCVs of rangeland cattle at high elevation. We specifically wanted to determine if Hct/PCV levels measured with a commercial PoC instrument (i-Stat with CHEM8+ cartridges [PoCi ]) were comparable to Hct/PCV levels measured with traditional laboratory methods. We assessed the Hct/PCV of 94 mature beef cattle (black Angus; Bos taurus) at ~2195m above sea level using paired analyses of the PoCi and TCC methods from each animal. We used paired samples t-tests to compare mean Hct/PCVs. Correlation analyses relative to the line of identity and Passing-Bablok regression were used to assess systematic and proportional differences, respectively, and Bland-Altman plots were used to assess agreement between the two methods. The PoCi estimated a Hct of 28.2% ± 0.7% (SE), which was lower than the TCC estimated PCV of 39.2% ± 0.5%. The Bland-Altman plot revealed poor agreement between the two methods in addition to a -11% bias for the PoCi . The Passing-Bablok regression revealed both systematic and proportional bias between the two methods. Point-of-care blood instruments were not comparable to TCC methods for quantifying Hct/PCVs of cattle living at high elevations.

Macronutrient, immunoglobulin a and total antioxidant capacity profiles of human milk from 1 to 24\u2009months: a cross-sectional study in Thailand

BackgroundAn extended duration of breastfeeding of up to two years is encouraged by many health authorities, but information regarding the composition of milk after one year postpartum is limited. The goal of this study was to determine the association between the duration of lactation and macronutrient contents, immunoglobulin A (IgA) levels and total antioxidant capacity (TAC) in human milk (HM), from 1 to 24 months postpartum.MethodsCross-sectional milk samples were collected between January and April 2019 from mothers with healthy full-term children who had been lactating for 1 to 24 months. The HM was biochemically analyzed for protein and carbohydrate contents by colorimetric assays. The fat content was determined by capillary centrifugation, and the energy content was calculated from the results of centrifugation assays. IgA levels and TAC were determined by ELISA and a Trolox equivalent antioxidant capacity (TEAC) assay, respectively. Pearson’s correlation coefficient and Spearman’s rank correlation coefficient were used to determine associations between months of lactation and milk composition, and multiple regression analysis was used to assess associations between months of lactation and milk composition adjusted for relevant covariates. Differences were considered significant at p < 0.05.ResultsOne hundred eighty-four milk samples were analyzed. The month of lactation was positively associated with the fat concentration (B = 0.31, SE = 0.09, p = 0.001), energy content (B = 3.11, SE = 0.92, p = 0.001), and IgA (B = 4.17, SE = 1.08, p < 0.001) but negatively associated with the carbohydrate concentration (B = − 0.22, SE = 0.01, p = 0.04). No association was observed between the month of lactation and the protein concentration or TAC after adjustment for maternal age, maternal BMI, birth order, and breastfeeding frequency.ConclusionThe duration of lactation was found to be positively associated with the fat, energy, and IgA content in HM for up to two years postpartum, and negatively associated with carbohydrate concentration. More prospective cohort studies are needed to obtain evidence-based knowledge regarding the changes in HM composition throughout the course of lactation.

Serological and molecular analysis of ABO and Rh blood group chimeras

The serological examination, blood transfusion strategies and the molecular analysis to blood group chimera were conducted to demonstrate existent of chimera in blood group. The blood grouping of ABO or/and RhD, newborn red blood cells separated by capillary centrifugation. Aabsorption tests and DTT treated agglutination erythrocyte tests were implemented in four patients. Further molecular biological research was conducted on one patient's sample. The results showed that for patient 1: ABO blood group was AB/B chimera, Rh blood cells contained the RhCE chimera gene; Patient 2: Rh blood cells contained the RhD chimera gene; Patient 3: ABO blood group was AB/B chimera, Rh blood cells contained the RhD chimera gene; Patient 4: ABO blood group was O/B chimera, Rh blood cells contained the RhCE chimera gene. The study suggests that the individuals categorized as chimeras are likely to be more common than existing literature reports. According to the serological tests, in the absence of a history of recent blood transfusion or disease to cause reduced antigen, the phenomena of hybrid aggregation of the ABO and Rh blood system were the main feature. In terms of transfusion strategy, the selection of ABO and Rh blood groups should be depended on the group of cells with more antigens.

Identification of RBC and WBC Count in Human Blood Using ARM Based Instrumentation

The rapid growth in microelectronics and crunching RISC in the field of bio-medical sciences incorporated of soft tools to diagnose various parameters of human fluids. Conventional method of blood sample analysis makes use of laboratory technique of titration, which is operator-dependent and results in lot of errors depending on the skill of the technician. In order to eliminate the human errors involved in the conventional method, in this paper an attempt has been made to present a capillary centrifuge technique driven by high speed DC motor fed by Morgan chopper and controlled by powerful ARM processor. It results in accurate analysis of the blood samples. The various techniques involved in accurate sensing of speed using timer and generation of firing pulses to thyristor in the Morgan chopper is judiciously achieved. This paper clearly brings out the advantages of the proposed blood measurement technique which effectively gives blood analysis faster and at a low cost.

Integration of diagnosis and treatment of sleeping sickness in primary healthcare facilities in the Democratic Republic of the Congo.

Control of human African trypanosomiasis (HAT) in the Democratic Republic of Congo (DRC) has always been a vertical programme, although attempts at integration in general health services were made in recent years. Now that HAT prevalence is declining, the integration question becomes even more crucial. We studied the level of attainment of integration of HAT case detection and management in primary care centres in two high-prevalence districts in the province of Bandundu, DRC. We visited all 43 first-line health centres of Mushie and Kwamouth districts, conducted structured interviews and inspected facilities using a standardised checklist. We focused on: availability of well trained staff - besides HAT, we also tested for knowledge on tuberculosis; availability of equipment, consumables and supplies; and utilisation of the services. All health centres were operating but most were poorly equipped, and attendance rates were very low. We observed a median of 14 outpatient consultations per facility (IQR 8-21) in the week prior to our visit, that is two patients per day. The staff had good knowledge on presenting symptoms, diagnosis and treatment of both HAT and tuberculosis. Nine centres were accredited by the national programme as HAT diagnosis and treatment centres, but the most sensitive diagnostic confirmation test, the mini-anion exchange centrifugation technique (mAECT), was not present in any. Although all nine were performing the CATT screening test, only two had the required cold chain in working order. In these high-prevalence districts in DRC, staff is well-acquainted with HAT but lack the tools required for an adequate diagnostic procedure. Attendance rates of these primary care centres are extremely low, making timely recognition of a resurgence of HAT unlikely in the current state of affairs.

Performance of parasitological and molecular techniques for the diagnosis and surveillance of gambiense sleeping sickness.

ObjectivesRecently, improvements have been made to diagnostics for gambiense sleeping sickness control but their performance remains poorly documented and may depend on specimen processing prior to examination. In a prospective study in the Democratic Republic of the Congo, we compared the diagnostic performance of several parasite detection techniques, immune trypanolysis and of m18S PCR on whole blood stored in a stabilisation buffer or dried on filter paper.MethodsIndividuals with CATT whole blood (WB) titer ≥1∶4 or with clinical signs indicative for sleeping sickness were examined for presence of trypanosomes in lymph node aspirate (LNA) and/or in blood. Blood was examined with Capillary Centrifugation Technique (CTC), mini-Anion Exchange Centrifugation Technique (mAECT) and mAECT on buffy coat (BC). PCR was performed on whole blood (i) stored in guanidine hydrochloride EDTA (GE) stabilisation buffer and (ii) dried on filter paper, and repeatability and reproducibility were assessed. Immune trypanolysis (TL) was performed on plasma.ResultsA total of 237 persons were included. Among 143 parasitologically confirmed cases, 85.3% had a CATT-WB titre of ≥1/8, 39.2% were positive in LNA, 47.5% in CTC, 80.4% in mAECT-WB, 90.9% in mAECT-BC, 95.1% in TL and up to 89.5% in PCR on GE-stabilised blood. PCR on GE-stabilised blood showed highest repeatability (87.8%) and inter-laboratory reproducibility (86.9%). Of the 94 non-confirmed suspects, respectively 39.4% and 23.4% were TL or PCR positive. Suboptimal specificity of PCR and TL was also suggested by latent class analysis.ConclusionThe combination of LNA examination with mAECT-BC offered excellent diagnostic sensitivity. For PCR, storage of blood in stabilisation buffer is to be preferred over filter paper. TL as well as PCR are useful for remote diagnosis but are not more sensitive than mAECT-BC. For TL and PCR, the specificity, and thus usefulness for management of non-confirmed suspects remain to be determined.

Design of a Novel Blood Cell Analysis System Based on Capillary Centrifugation and Stratification Technology

The test of complete blood cells plays a significant role in diagnosis and treatment of diseases. In order to take an immediate quantitatively measurement of red blood cells, platelets, lymphocytes, monocytes and granulocytes, as well as to realize the binary classification of white blood cells, we design a novel blood cell analysis system based on capillary centrifugation and stratification technology. Blue and red LED array are adopted as light sources, and one-dimensional images of the capillary can be got from the color linear CCD. By further analysis can we obtain the final nine parameters. In the paper, we mainly focus on description of the system’s working principle and the general design. Experimental results show that our system is of high accuracy and good repeatability.

Trypanosoma evansi: A clinical, parasitological and immunological evaluation of trypanosomosis using a chronic rabbit model

We evaluated the clinical, parasitological and immunological effects of a Venezuelan strain of Trypanosoma evansi (T. evansi) throughout in experimentally inoculated rabbits over the course of infection and compared them with the same aspect in healthy animals. Body temperature was recorded in degrees Celsius, animal weight in kilograms, serum proteins in g/dl using a refractometer, haematocrit percentage by capillary centrifugation and the anti-T. evansi IgG titer by indirect ELISA immunoassay, from both infected animals and controls for 95 days. Infected animals showed a higher body temperature, total serum protein and anti- T. evansi antibody titer, and a lower haematocrit and weight gain than controls. These differences were related to the presence of the parasites in the blood as detected micro-haematocrit centrifugation technique (MHCT) and direct microscopic examination (DME). This study confirms the usefulness of rabbits as a model for the study of trypanosomosis; the clinical features of the disease can be observed and the three characteristic stages, prepatent period, acute and chronic phase clearly defined over the course of the infection

Cytologic Examination of Hemorrhagic Fluid by Capillary Centrifugation

To develop a simplified technique for processing hemorrhagic body fluids, allowing elimination of red blood cells (RBCs) to obtain tumor cell-rich cytosmears for accurate diagnosis of malignancy.Hemorrhagic fluid is collected with ethylenediamine tetraacetic acid anticoagulant. The cells are separated by centrifugation in glass capillaries into 3 layers, with the uppermost containing supernatant, middle buffy coat and lowermost dark layer of RBCs. The smears of the buffy coat are prepared by breaking the capillaries at the junction of buffy coat and RBC layer. The procedure is named capillary centrifugation technique. This procedure was developed in the cytology laboratory at King George's Medical College, Lucknow, India in 1974.Cell yield is good and cellular details are well preserved. No cell debris is present on the slides. The background of the smears is absolutely clear. Any number of slides may be prepared for special study for exact typing of tumor cellsThis is a simple, economical procedure and a good substitute for a cytocentrifuge machine for preparing smears from small amount of fluids. Technicians learn it quickly and are quite comfortable with the procedure.

Adolescents and HIV/AIDS: an update of cases seen in Benin City, Nigeria

The seroprevalence of HIV infections was investigated among 1,208 adolescents aged 13–19 years who presented for HIV testing between 1994 and 2000 at the two largest hospitals in Benin City, Edo State, Nigeria. The serum samples of the subjects were tested for HIV antibodies using ELISA and/or rapid test kits and confirmed by Western blot. The overall prevalence rate was high (11.4%), with a higher percentage (74.6%) being recorded in females as compared to males. There was a marked increase in prevalence from 7.0% in 1995 to 13.3% in 2000. Studies in Nigeria have shown that adolescents are sexually active, have multiple sexual partners and have recorded high prevalence of STDs. In view of the association of HIV infection with these high risk factors, the results of this study further re-echoes the urgent need to target specific intervention programmes at adolescents. Key words: HIV/AIDS, adolescents, Benin City, Nigeria [Jnl Medicine & Biomedical Res. Vol.1(2) 2002: 59-62] Haemorheological parameters in hypertensive Nigerians compared to normotensive controls DV Dappera and DA Ighorojeb aDepartment of Physiology, College of Health Sciences, University of Port Harcourt, P. M. B. 5323, Port Harcourt, Nigeria. E-mail: dapperdv2001@yahoo.com bDepartment of Physiology, College of Medical Sciences, University of Benin, P. M. B. 1156, Benin City, Nigeria. Tel: 052 600781 E-mail: ahboruniben.edu ; dighoroje@yahoo.com ; dighoroje@hotmail.com Correspondence: Dr A. D. A. Ighoroje, Department of physiology, College of Medical Sciences, University of Benin, P. M. B. 1156, Benin City, Nigeria. Tel: 052 600781 E-mail: ahboruniben.edu ; dighoroje@yahoo.com ; dighoroje@hotmail.com : The values of five important haemorheological parameters, namely, haematocrit (Hct), erythrocyte sedimentation rate (ESR), haemoglobin concentration (Hbc), fibrinogen concentration (FC), relative plasma viscosity (RPV) and whole blood relative viscosity (WBRV) were determined in 30 male and 20 female hypertensives without complications, aged between 30 and 70 years. Thirty male and 20 female normotensives, matched for age, weight and height served as controls. Hct was determined by Hawksley micro capillary centrifugation, Hbc by the cyanmethaemoglobin method, FC by the clot weight method, ESR by the Westergren method, while both RPV and WBRV were determined by capillary viscometry. Systolic and diastolic blood pressures were measured by standard clinical procedures and mean arterial pressure (MAP) determined. The results show significantly higher values of FC, RPV and WBRV in the hypertensives (p < 0.05). By contrast, there were no significant differences in ESR, Hbc and Hct between the hypertensive and normotensive subjects. We conclude that the increases in the various haemorheological parameters in Nigerian hypertensives are attributable to the significantly elevated levels of fibrinogen in hypertensive subjects. Acceptability of surgery as initial treatment for primary open angle glaucoma AE Omoti, OT Edema and MJM Waziri-Erameh Department of Ophthalmology, University of Benin Teaching Hospital, Benin City. Correspondence: AE Omoti, Department of Ophthalmology, University of Benin Teaching Hospital, Benin City. : One hundred and fifty four new glaucoma patients at the University of Benin Teaching Hospital were interviewed to determine the acceptance of surgery, reasons for refusal, and factors affecting acceptability of surgery as an initial treatment option. Fifty patients (32.5%) accepted surgery while 89 (57.8%) refused. The main reasons for refusal were knowledge that there may be no visual improvement after surgery [22 patients (24.7%)], fear of surgery [22 patients (24.7%)] and cost of surgery [15 patients (16.9%)]. Very poor vision was significantly associated with refusal of surgery (c2 = 21.72, df = 10, p = 0.016) while hospital or clinic attendance prior to presentation was significantly associated with acceptance of surgery (c2 = 24.52, df = 8, p = 0.0019). This study shows that the level of acceptance of surgery as initial treatment for glaucoma by patients is low. Public enlightenment campaigns on the safety and efficacy of surgery need to be embarked upon. Key words: Acceptability, surgery, glaucoma

Haemorheological parameters in hypertensive Nigerians compared to normotensive controls

The values of five important haemorheological parameters, namely, haematocrit (Hct), erythrocyte sedimentation rate (ESR), haemoglobin concentration (Hbc), fibrinogen concentration (FC), relative plasma viscosity (RPV) and whole blood relative viscosity (WBRV) were determined in 30 male and 20 female hypertensives without complications, aged between 30 and 70 years. Thirty male and 20 female normotensives, matched for age, weight and height served as controls. Hct was determined by Hawksley micro capillary centrifugation, Hbc by the cyanmethaemoglobin method, FC by the clot weight method, ESR by the Westergren method, while both RPV and WBRV were determined by capillary viscometry. Systolic and diastolic blood pressures were measured by standard clinical procedures and mean arterial pressure (MAP) determined. The results show significantly higher values of FC, RPV and WBRV in the hypertensives (p < 0.05). By contrast, there were no significant differences in ESR, Hbc and Hct between the hypertensive and normotensive subjects. We conclude that the increases in the various haemorheological parameters in Nigerian hypertensives are attributable to the significantly elevated levels of fibrinogen in hypertensive subjects. [Jnl Medicine & Biomedical Res. Vol.1(1) 2002: 63-67]

A Retrospective Study of Glucose Metabolism in Mothers of Large Babies

To evaluate retrospectively the glucose metabolism of women who gave birth to large babies. Within the first 3 days of postpartum, HbA1c of the dense erythrocytes and glycated albumin were measured. The sample group was comprised of 59 women who gave birth to heavy-for-dates infants. Fifty-five women who had average-sized infants formed the control group. Both groups had normal results on screening for gestational diabetes mellitus (GDM) by 28 weeks of gestation. The dense erythrocyte fractionation was obtained by capillary centrifugation for measurement of stable HbA1c. Mothers of heavy-for-dates infants had a significantly higher level of HbA1c of the dense erythrocytes (mean +/- SD; 5.86 +/- 0.68 vs. 5.50 +/- 0.70%, P = 0.015) and a higher maternal body mass index (BMI) (mean +/- SD; 26.5 +/- 2.9 vs. 24.1 +/- 2.3 kg/m2, P < 0.00001) than control mothers. Differences in the occurrence of high levels of HbA1c of the dense erythrocytes and maternal BMI were statistically significant (P < 0.01, P < 0.05) between the two groups, but no difference was noted in glycated albumin. There was no statistical correlation between high levels of HbA1c of the dense erythrocytes and maternal obesity. High HbA1c levels of the dense erythrocytes and maternal obesity are independent factors that affect fetal oversize. Heavy-for-dates infants may be a consequence of maternal hyperglycemia in late pregnancy. This maternal hyperglycemia was not detected by the routine screening test for GDM.

Glycated hemoglobin of fractionated erythrocytes, glycated albumin, and plasma fructosamine during pregnancy

OBJECTIVE: To evaluate glucose metabolism during pregnancy, we measured plasma fructosamine, glycated albumin, and the stable glycated hemoglobin of the light and dense erythrocytes. STUDY DESIGN: The abnormal glucose tolerance group comprised patients with gestational diabetes and those with one abnormal value on a 75 gm oral glucose tolerance test. Erythrocyte fractionation was performed by capillary centrifugation. RESULTS: In normal pregnancy glycated hemoglobin of the light erythrocytes was reduced in the second and third trimesters (3.42% ± 0.62% [mean ± SD] [ n = 306] in the first trimester, 2.15% ± 0.48% [ n = 353] in the second, and 2.06% ± 0.58% [ n = 300] in the third), and dense erythrocytes were higher in the third trimester (first 4.59% ± 0.46%, second 4.70% ± 0.49%, third 5.29% ± 0.73%). Glycated albumin and fructosamine followed a pattern similar to the light erythrocytes. The group with abnormal glucose tolerance had significantly higher levels of glycated hemoglobin of the light erythrocytes in the first and third trimesters and glycated hemoglobin of the dense erythrocytes and glycated albumin in all trimesters. CONCLUSION: The biphasic change in nonfractionated glycated hemoglobin is the sum of the lower glycated hemoglobin of the light erythrocytes and the higher glycated hemoglobin of the dense erythrocytes in late pregnancy. The stable glycated hemoglobin of fractionated erythrocytes and the glycated albumin accurately reflect maternal glucose metabolism during pregnancy.

Simplified modification of the macrophage migration inhibition method in mice

Simplified modification of the macrophage migration inhibition method is suggested. The simplification is achieved by omitting the capillary centrifugation stage.

Memory and Transfer of Information

Memory and Transfer of Information

Capillary centrifugation.

Capillary centrifugation.

Simplified technique of high-speed capillary centrifugation

Simplified technique of high-speed capillary centrifugation