Hematological alterations and thymic function in newborns of HIV-infected mothers receiving antiretroviral drugs

Objectives

To investigate the effects of antiretroviral (ARV) drugs on hematological parameters and thymic function in HIVuninfected newborns of HIV-infected mothers.

Study design

Cross sectional study.

Setting

Chiang-Mai University Hospital, Chiang-Mai, Thailand.

Participants/Patients

49 HIV-uninfected and 26 HIV-infected pregnancies.

Methods

Cord blood samples of newborns from HIV-uninfected and HIV-infected mothers were collected. Hematological parameters were measured using automatic blood cell count. T-cell receptor excision circles (TRECs) levels in cord blood mononuclear cells (CBMCs), CD4⁺ and CD8⁺ T-cells were quantified using real-time PCR.

Main Outcome Measures

Hemotological parameters and thymic function.

Results

Newborn of HIV-infected mother tended to have lower mean levels of hemoglobin than those of HIV-uninfected mother (137 ± 22 vs 146 ± 17 g/L, P = 0.05). Furthermore, mean of red blood cell (RBC) counts and hematocrit and median of TRECs in CD4+ T-cells in the newborns of the former were significantly lower than those of the latter [3.6 ± 0.7 vs 4.8 ± 0.6 × 10₁₂ cells/L, P <0.001; 0.40 ± 0.07 vs 0.46 ± 0.05 L/L, P <0.001 and 0.53 (IQR: 0.03–5.76) vs 13.20 (IQR: 2.77–27.51) × 10^?3 pg/μL, P = 0.02, respectively].

Conclusion

ARV drugs altered hematological parameters and thymic function (TRECs CD4⁺ T-cells) in HIV-uninfected newborns of HIV-infected mothers.     

Detection of alpha-thalassemia-1 Southeast Asian type using real-time gap-PCR with SYBR Green1 and high resolution melting analysis

Alpha-thalassemia-1 Southeast Asian (SEA) type is the most common genetic disorder in the Asian population. Couples who are both carriers have a 25% chance of conceiving Bart's hydrops fetalis. Therefore, results from carrier screening and prenatal diagnosis frequently need to be available rapidly. A rapid technique for diagnosis of alpha-thalassemia-1 SEA type was implemented. The technique used is based on real-time gap-PCR and high resolution melting (HRM) analysis of the amplified fragment using the Rotor-Gene 6000. The DNA samples used for amplification were obtained from whole blood, cord blood, and chorionic villus sampling (CVS). With this method, the alpha-thalassemia-1 SEA allele can be easily distinguished from wild type alpha-globin gene allele. The real-time gap-PCR and HRM analysis offers additional benefits including minimal labor, rapid turnaround time, and a decreased risk of PCR carryover contamination. It is cost-effective and safe, does not require fluorescently labeled probe and hazardous chemicals. Moreover, it is accurate showing 100% concordance with conventional gap-PCR analysis.     

Evaluation of the placental environment with a new in vitro model of histocultures of early and term placentae: determination of cytokine and chemokine expression profiles

We aimed to set up and validate a new in vitro model of placental histocultures, for the evaluation of cytokine and chemokine profiles of the placental environment, over a long culture period. Micro-explant cultures from 6 early and 6 term placentae were set up on collagen sponge gel supports at a liquid/air interface. At various times during culture, we analyzed tissue morphology and cell death by microscopy and quantified beta-hCG production and mRNA levels for beta-hCG and insulin-like 4 (INSL4). Levels of IL-6, LIF, TNF alpha, IL-10, IFN-gamma, IL-16 and RANTES in the medium were measured by ELISA on days 1, 4 and 7 of culture. SDF-1 mRNA expression was determined by real-time PCR at the same time points. Histocultures from early and term placentae remained viable until day 10. High levels of IL-6 and LIF production, low levels of TNF alpha, IL-10 and IFN-gamma production and significant SDF-1 expression were observed. These data indicate that placental histoculture is a suitable and reliable in vitro model for studying the placental environment.     

HbE Level and Red Cell Parameters in Heterozygous HbE With and Without α<Superscript>0</Superscript>-Thalassemia Trait

We compared hemoglobin (Hb) E levels and red cell parameters between heterozygous HbE with and without α⁰-thalassemia trait and also determine their appropriated cut-off points for differentiating these two groups. High performance liquid chromatography analysis results and mean levels of red blood cell (RBC) parameters, including RBC count, total Hb, hematocrit, MCV, MCH and MCHC of heterozygous HbE with α⁰-thalassemia trait (n?=?183) and without α⁰-thalassemia trait (n?=?1437) were reviewed and compared. The α⁰-thalassemia status in these samples was detected by real-time PCR with SYBR Green1 and high resolution melting analysis. Mean levels of HbE, total Hb, MCV, MCH and MCHC of heterozygous HbE with α⁰-thalassemia trait were significantly lower than those of heterozygous HbE without α⁰-thalassemia trait (P?<?0.001). In addition, HbE level at a cut-off value of?<?24% was superior to MCV (<?80 fL) and MCH (<?27 pg) for differentiating the heterozygous HbE with and without α⁰-thalassemia trait with 100% sensitivity and 87.2% specificity. Despite certain limitations of this study like missing RDW and reticulocyte counts, and not testing for α⁺-thalassemia and Hb Constant Spring, we conclude that the HbE level at a cut-off point of?<?24% is a useful marker for initial discrimination between heterozygous HbE with and without α⁰-thalassemia trait.     

Interference of hemoglobin Hope on beta-thalassemia diagnosis by the capillary electrophoresis Method

The β-chain hemoglobin (Hb) variants interfere with the diagnosis of β-thalassemia trait using high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE). We analyzed the effect of Hb Hope, a β-chain Hb variant frequently found in the Thai population, on β-thalassemia trait diagnosis. HPLC and CE were used to quantify the level of HbA(2) in 11 whole blood samples containing Hb Hope. The levels of Hb Hope detected by both methods were similar. An elevated HbA(2) level was found in all samples analyzed by the CE method, while 1 was increased when analyzed by HPLC, which was a compound heterozygous of Hb Hope and α-thalassemia-1 SEA-type deletion. Of 11 samples, 6 had mean corpuscular volumes within the reference range. All samples showed negative results for molecular analysis of β(0)-thalassemia codon 17, 41/42, and 71/72 mutations and β-thalassemia 3.5-kb deletion. Therefore, Hb Hope interfered with the diagnosis of β-thalassemia trait analyzed by CE but not by HPLC.     

Health symptoms related to pesticide exposure and agricultural tasks among rice farmers from northern Thailand

Purpose

The objective of this cross-sectional study was to investigate health symptoms related to occupational pesticide exposure and agricultural tasks in rice farmers.

Methods

Data on demographic variables and health symptoms associated with pesticide exposure were collected from 182 rice farmers (exposed subjects) and 122 non-farmers (controlled group) using interviews and measuring whole blood acetylcholinesterase (AChE) activity during August and October 2012.

Results

Rice farmers had a significantly lower median AChE activity than the controls (9,594 vs. 10,530 U/L, respectively) and a significantly higher prevalence of difficulty in breathing and chest pain [odds ratio (OR) 2.8, P < 0.01 and OR 2.5, P < 0.05, respectively]. The prevalence of dry throat and cramp was associated with those farmers who sprayed and mixed pesticides (OR 2.5 and 2.6 for dry throat, OR 2.5 and 2.9 for cramp, respectively; P < 0.01). The prevalence of numbness and diarrhea was associated with those farmers who scattered seed (OR 2.2, P < 0.01 and OR 3.6, P < 0.05, respectively). The prevalence of numbness and increasing anxiety was also associated with those farmers who harvested crops (OR 3.6, P < 0.01 and OR 3.0, P < 0.05, respectively).

Conclusions

Our findings suggest that occupational pesticide exposure and agricultural tasks in the paddy field may be associated with the increasing prevalence of respiratory tract and muscle symptoms. This possibility warrants further investigation in more detail.     

Comparison of capillary electrophoregram among heterozygous Hb Hope, Hb Hope/α-thalassemia-1 SEA type deletion and Hb Hope/β0-thalassemia

Abstract Background: Hemoglobin (Hb) A2 is artifactually elevated in cases of heterozygous Hb Hope when measured by capillary electrophoresis (CE). However, there is no report of HbA2 levels and capillary electrophoregrams for associations of heterozygote of Hb Hope with α-thalassemia nor β-thalassemia. Methods: Levels of HbA0, HbA2 and Hb Hope in 16 heterozygous Hb Hope, 3 Hb Hope/α-thalassemia-1 SEA type deletion and 2 Hb Hope/β0-thalassemia were measured by CE. Electrophoregram and the levels of those were compared within these three groups. Results: Artifactually elevated HbA2 levels (≥4%) were found in both groups of heterozygous Hb Hope and Hb Hope/α-thalassemia-1 SEA type deletion. Manual corrections were performed by adjusting baselines, and results showed that means of HbA2 in both groups decreased from 4.47% and 4.03% to 1.93% and 1.77%, respectively. The highest levels of HbA2 and Hb Hope were observed in samples with Hb Hope/β0-thalassemia. Moreover, HbA0 was not observed in these cases. Conclusions: The elevation of HbA2 in patients with heterozygous Hb Hope and with Hb Hope/α-thalassemia-1 SEA type deletion measured by CE leads to incorrect β-thalassemia trait diagnosis. However, using CE electrophoregram together with levels of HbA0, HbA2 and Hb Hope would be a more accurate and precise method for diagnosis of Hb Hope/β0-thalassemia.