A Simple Whole-Blood Polymerase Chain Reaction without DNA Extraction for Thalassemia Diagnosis

Abstract

Polymerase chain reaction (PCR) diagnosis of thalassemia usually relies on using genomic DNA. Preparing the genomic DNA can lead to sample-to-sample contamination. This report was aimed to establish the PCR protocol using whole-blood for detecting mutations of α- and β-globin genes causing the thalassemia syndrome. First, the PCR facilitators, betaine and bovine serum albumin (BSA), were tested, simultaneously with an adjustment of PCR thermal cycler and of whole-blood volume. Thereafter, the established whole-blood PCR was applied for detecting, in both known and unknown samples, the HBA1 Southeast Asian (–?–^SEA) (NG_000006.1: g.26264_45564del19301) deletion, Hb Constant Spring (Hb CS, HBA2: c.427T>C, α^CSα), codon 17 (A>T) (HBB: c.52A>T), codons 41/42 (–TTCT) (HBB: c.126_129delCTTT) deletion, –28 (A>G) (HBB: c.-78A>G) and codon 26 (G>A) (Hb E or HBB: c.79G>A). It was shown that the whole-blood PCR worked successfully in 9.0% (w/v) betaine, with 1?μL of EDTA whole blood and with addition of 10 heat-cool steps (3?min. at 94?°C, followed by 3?min. at 55?°C) prior to the typical thermal cycles for the mutations. The capability of the new whole-blood PCR was similar to that of the typical DNA-based PCR. Therefore, the newly established whole-blood PCR could be performed for PCR diagnosis of thalassemia. Using this platform, sample-to-sample contamination should be eliminated.     

Adenomatoid odontogenic tumor mimicking a periapical (radicular) cyst: a case report

The adenomatoid odontogenic tumor (AOT) is a benign (hamartomatous), noninvasive lesion with slow but progressive growth. A rare subvariant of the extrafollicular type of AOT may mimic periapical disease radiographically. We report on a 15-year-old girl with a periapical radiolucent lesion affecting teeth 7 and 8 initially suspicious of periapical disease, although clinical findings seemed to indicate a nonendodontic lesion. An exploratory surgical approach was chosen, and the final diagnosis of the removed noncystic tissue mass was microscopically confirmed to be an AOT.     

Radiologic findings in tsunami trauma: experience with 225 patients injured in the 2004 tsunami

The objective of this study was to determine the spectrum of radiographic findings, frequency, and type of injuries in tsunami victims. From December 2004 to May 2005, all tsunami victims admitted to our hospital were retrospectively identified by a search of medical records. Patients who received radiologic examinations were reviewed for their radiographic findings. The authors identified 225 tsunami victims. One hundred eight victims received radiologic evaluations on admission that included 350 plain radiographs, 19 ultrasound exams, 18 computed tomography (CT) scans and 3 magnetic resonance imaging (MRI) scans. Overall positivity rate was 48% (187/390). Most common trauma involved musculoskeleton (102/187, 54.5%). Retained foreign bodies in soft tissues, pneumonia/aspiration, and tsunami sinusitis were found in 22, 28, and 31 patients (9.8, 12.4, and 13.8%), respectively. These were unique findings in tsunami trauma. Imaging played an important role in detection of these abnormalities.     

Physical mapping of rDNA and heterochromatin in chromosomes of 16 Coffea species: A revised view of species differentiation

The chromosome organization among 15 wild diploid Coffea species and cultivated tetraploid C. arabica was determined by fluorochrome banding (CMA, DAPI) and double fluorescence in-situ hybridization (FISH) of 5S and 18S rDNA achieved on the same chromosome plates. Two to five chromosome pairs (plus one putative chromosome B) are marked. Overall, there are two SAT-chromosome pairs for East African species and one for the Malagasy and the West and Central African species. 18S rDNA loci are telomeric and strongly marked the SAT-chromosome pairs. Generally, only one pericentromeric 5S rDNA locus characterized East African species, while an additional minor locus co-localized with the 18S rDNA-SAT locus for the Malagasy species and West and Central African species. A combination of rDNA FISH plus CMA and DAPI banding patterns enables identification of almost all the species, even those for which the genetic or botanical status is still being discussed. C. arabica clearly appears to be an allotetraploid species, including one genome from East Africa and one from West and Central Africa. However, since the minor 5S rDNA-SAT locus present in West/Central African genomes is not detected, two evolutionary hypotheses could be put forward for C. arabica. Considering only the diploid species, global trends are obvious in rDNA signal patterns, genome size variations, and geographic distribution of the species, but there are no clear evolutionary trends. However, complex interactions between these factors and environmental growing conditions exist, which have resulted in loss and gain of rDNA loci and probably also in copy repeat number variations in each rDNA family.     

Dynamic Irrigation Promotes Apical Papilla Cell Attachment in an Ex Vivo Immature Root Canal Model

Introduction

The use of dentin preconditioning techniques in regenerative endodontic procedures is currently promising. Several growth factors have been detected on dentin after ultrasonic irrigation with EDTA. This study aimed to evaluate the effects of dynamic irrigation with different solution regimens on apical papilla cell (APC) attachment in an ex vivo immature tooth model.

Idiopathic high-flow priapism in a pediatric patient

High-flow priapism is a very rare condition in children. The most common cause is perineal trauma, which is a major cause of arterio-cavernosal fistula. A few pediatric patients have high-flow priapism without an obvious cause. There are many therapeutic modalities for this condition, depending on the etiology. We report a case of idiopathic high-flow priapism in a 6-year-old boy who underwent repeated superselective embolization.     

Pediatric renovascular hypertension in Thailand: CT angiographic findings

Background  

Renovascular disease is an uncommon but important cause of hypertension in children. When unrecognized and untreated, renovascular hypertension in children can have serious complications.     

Outcomes of Direct Pulp Capping by Using Either ProRoot Mineral Trioxide Aggregate or Biodentine in Permanent Teeth with Carious Pulp Exposure in 6- to 18-Year-Old Patients: A Randomized Controlled Trial

Introduction

This study aimed to compare the success rates of direct pulp capping (DPC) by using either ProRoot Mineral Trioxide Aggregate (MTA) or Biodentine in the cariously exposed permanent teeth of 6- to 18-year-old patients. Gray discoloration was also evaluated.