A simple and rapid polymerase chain reaction-based method for detecting a prevalent mutation (R413P) in Japanese phenylketonuria patients

A convenient molecular method for the detection of R413P (¹²³⁸G←C) mutation in exon 12 of the phenylalanine hydroxylase gene, one of the prevalent mutations among Japanese patients with classical phenylketonuria (PKU) is described. The mutation was previously detected by polymerase chain reaction (PCR)-direct sequencing or allele specific oligonucleotide hybridization. However, these methods were cumbersome and only a few laboratories could provide such a diagnostic service. An improved version of the method has been developed here, involving 30 cycles of PCR following restriction enzyme digestion. In the upstream primer encompassing G-1207 to C-1237, two substitutions are artificially introduced, so that a Bam-HI site involving C-1238 is introduced in the copies of the mutant allele. With the use of this method, R413P-homozygote and -heterozygote can be readily and unequivocally distinguished from normal using genomic DNA extracted from peripheral blood leukocytes. Among 10 Japanese PKU patients investigated, three were homozygous and three were heterozygous for the R413P allele, whereas four did not carry this mutant allele, indicating that the prevalence of the mutant allele is 45%. The result suggests that it is technically feasible to develop a program for carrier detection of the mutation in the Japanese population.     

Neurally mediated syncope complicated with paroxysmal atrial fibrillation

Recently, it has come to be accepted that the result of the tilt test is specific to neurally mediated syncope (vasovagal syncope). Only rarely is a case of paroxysmal atrial fibrillation without any organic diseases in childhood reported. A case reported here of a 14-year-old boy with neurally mediated syncope; which was complicated by paroxysmal atrial fibrillation, and which was diagnosed by performing the tilt test. Atrial fibrillation can be induced by the extraordinary stimulation of the vagal nerve during syncope. In a child, neurally mediated syncope complicated with paroxysmal atrial fibrillation has not been previously reported.     

Primary retroperitoneal lymphocele in a pregnant woman: The first report

In a 32-year-old pregnant woman, routine ultrasonography revealed right hydronephrosis and a huge retroperitoneal mass (20 x 7 cm) containing a fluid collection. Percutaneous drainage of the mass was performed and 2 L of clear, yellowish fluid was collected. Four months following the delivery, a recurrent retroperitoneal lymphocele was identified. Six months after the delivery, laparoscopic marsupialization was performed through a 10-mm umbilical camera port and two 5-mm ports on the right side of the abdomen. A posterior peritoneal window was established by creating a wide opening in the anterior wall of the lymphocele. Subsequent ultrasonography did not indicate a recurrence of the lymphocele or right hydronephrosis over a follow-up period of 8 months.     

Clinical Experiences with Laparoscopic Colectomy

Abstract: The indications for laparoscopic colon resection and the associated complications are discussed herein. This procedure was indicated for patients with benign disease or malignant disease with invasion limited to the submucosal layer. The subjects consisted of 14 cases with submucosal tumor invasion diagnosed preoperative, three with submucosal invasion clarified by endoscopic polypectomy, three with adenomas larger than 2 cm in diameter strongly suspected of being focal submucosal tumor invasion considered unsuitable for endoscopic mucosal resection and one with Crohn's disease with ileus. Two cases in whom laparoscopic surgery was not appropriate were included in this series. In one case with a superficial elevated lesion (Ma type), 15 mm in diameter, a diagnosis of moderately differentiated adenocarcinoma of the cecum was made preoperatively, but subserosal tumor invasion of the colonic wall with negative lymph node metastasis (nO) was revealed by examination of the resected specimen. The histology of the second superficial elevated lesion (Ha+lie type), which had a central depression, 13 mm in diameter and located above Bauhin's valve, was a well differentiated adenocarcinoma of which the cancerous portion invaded the proper muscle with positive lymph node metastasis (n1). Complications occurred in four cases. There were two cases of intraoperative vascular injury necessitating conversion to a standard laparotomy. One case with complete transection of the left ureter by End-GIA later underwent reoperation. The other case with minor leakage at the anastomotic site was managed with conservative therapy. In both of these cases the depth of tumor invasion had been incorrectly assessed as representing small elevated lesions, 15 mm in diameter, in the right colon. Furthermore, the cases who experienced complications had left colonic lesions. These results suggest that preoperative ultrasonic-endoscopy should be conducted as extensively as possible and that a good bloodless visual field appears to be necessary to avoid injuring adjacent organs.     

Case Report: Mucin-producing cystic neoplasm of the pancreas with onset in childhood

Mucin-producing tumours of the pancreas have been recently reported with increasing frequency and most cases have occurred in middle-aged and elderly people. In the present report, a case of a 21-year-old man with mucinous cystadenoma of the pancreas is reported. He had a long history of recurrent pancreatitis from the age of 8. When he was aged 10, the first branch of the main pancreatic duct was shown to be enlarged on endoscopic retrograde pancreaticography (ERP). A series of ERP studies and computed tomography scans performed over a period of 11 years demonstrated continuing growth of this enlargement of the pancreatic duct. Pancreaticoduodenectomy was performed and the patient has been well without further episodes of acute pancreatitis and has been free of recurrent tumour for 1 year.     

Use of Automatic Mode Change Between DDD and AAI to Facilitate Native Atrioventricular Conduction in Patients with Sick Sinus Syndrome or Transient Atrioventricular Block

The benefits of the automatic DDD (DDD/AMC) mode in the Chorus II pacemaker (Chorus 6234; Ela Medical Inc.), which automatically switches the modes between DDD and AAI to respect spontaneous AV conduction as much as possible in AAI while preserving safety pacing in DDD during paroxysmal AV block (AVB) only, remain unproven. This study examined the functions of the DDD/AMC mode in 12 patients with sick sinus syndrome (SSS; n = 10) or paroxysmal complete AVB (n = 2). A short-term (24 hours) comparative study between simple DDD mode and the DDD/AMC mode was performed in 8 of the 12 patients, and a medium-term (55.2 ± 54.6 days) follow-up of the DDD/AMC mode was completed in all 12 patients. A comparative pair of 24-hour surface Holter ECGs was obtained in 6 of the 8 patients in the short-term study. Telemetry and built-in Holter histograms were collected in the outpatient clinic in all 12 patients. Although the percentage atrial pacing of the telemetry increased from 59.2 ± 35.4 in DDD to 70.4 ± 31.8 in DDD/AMC (P < 0.009; n = 8), the percentage ventricular pacing decreased from 64.6 ± 37.7 in DDD to 36.2 ± 43.1 in DDD/AMC (P < 0.027) in the short-term study. In particular, the reduction of percentage ventricular pacing to < 10% was observed in four patients with SSS not associated with ± first-degree (1°) AVB on preoperative ECGs. Between the two modes a significant difference in arrhythmic events was not observed by the 24-hour surface Holter ECGS taken from the six patients in the short-term study. AAI-DDD switching associated with automatic modulation of AV delay and AV hysteresis occurred in all patients in the medium-term study. From the medium-term study, the total AV delay (AV delay plus AV hysteresis) exceeded 300 ms in 6 of the 12 patients in DDD/AMC, and usually became longest during nighttime. From the short- or medium-term study in the 12 patients, two patients preferred the DDD/AMC mode while one preferred the DDD mode. These results suggest that the DDD/AMC mode is useful, at least in SSS patients without ≥ 1° AVB, by reducing the percentage ventricular pacing.     

Distribution of Significant Changes in QRST Area Compared to Supraventricular Complex

To assess the effects of right ventricular (RV) pacing on body surface QRST distributions, we recorded QRST isointegral mops (I-maps) during sinus rhythm and RV pacing in 25 patients with anterior myocardial infarction (MI), 19 with inferior MI, and 14 without MI. The QRST values at each lead point recorded during sinus rhythm and RV pacing with an 87-lead system were analyzed with a paired t-test in each patient. An abnormal decrease in the QRST value of the I-map was assessed by the difference map, which indicated a −"2SD area," where the QRST integral value was less than the normal range (mean – 2SD) caiculated from 608 normal individuais. The I-maps were similar during the two activation sequences in patients with and without MI. However, during RV pacing, QHST values significantly decreased over the upper right anterior chest and increased over the lower left anterior chest and back. The ΣDMs (sum of QRST integral vaiues beiow the normal range) for both activation sequences were strongly correlated in patienis with anterior MI and with inferior MI (r = 0.91 and r = 0.92, respectively; P < 0.001). Although small but significant changes in QRST values were detected, the distribution of the "-2SD area" and the ΣDM were similar during both activation sequences in patients with prior MI. Thus, these findings demonstrate that an altered activation sequence produces small but significant changes in QRST values but that I-maps still provide information that is useful for the diagnosis of MI during RV pacing.     

Roles of specific cytokines in bone remodeling and hematopoiesis in Gaucher disease

BACKGROUND: Gaucher disease type 1 and type 3 are characterized by bone disease and hematological symptoms. It is known that monocyte/macrophage lineage is activated in Gaucher disease, and accordingly certain cytokines are elevated in blood. The aim of the present study was to explore the possible relationships between cytokines and bone remodeling and hematological abnormalities in this disease. METHODS: The concentrations of seven cytokines and two related proteins were measured in patients with Gaucher disease type 1 and type 3 (n= 8; age range, 2-50 years) who had received enzyme replacement therapy. RESULTS: Concentrations of interleukin-18 and transforming growth factor-beta1 were elevated in patients of all clinical types. Elevation of these cytokines in Gaucher disease has not been previously reported. Analysis of correlation among cytokines and bone-turnover markers showed that interleukin-18 concentration was correlated with each of two bone formation markers of bone-specific alkaline phosphatase activity and osteocalcin concentration, whereas macrophage colony-stimulating factor concentration correlated with the bone absorption marker of N-telopeptide to helix in urine. Concentrations of macrophage colony-stimulating factor and tumor necrosis factor-alpha were inversely correlated with hemoglobin concentration. CONCLUSIONS: Interleukin-18 and monocyte macrophage colony-stimulating factor are cytokines mainly involved in the mechanism of bone disease, while macrophage colony-stimulating factor and tumor necrosis factor-alpha may play a role in the development of hematological abnormalities in Gaucher disease.     

Epididymo-orchitis caused by intravesically instillated bacillus Calmette-Guérin: Genetically proven using a multiplex polymerase chain reaction method

The intravesical instillation of bacillus Calmette-Guérin (BCG) is a standard therapy for superficial bladder carcinoma. Tuberculosis-like inflammation in the genitourinary tract is a serious complication of BCG. It can occur after a long interval from the cessation of the intravesical BCG therapy. If inflammation occurs, it is necessary to test whether the BCG strain has caused it or another mycobacterium species has. However, there has never been a report that proves BCG causes the inflammation, because BCG is difficult to differentiate from other strains of Mycobacterium bovis and other members of the Mycobacterium tuberculosis complex by conventional tests, including regular polymerase chain reaction (PCR). We first present a case of epididymo-orchitis, which developed 31 months after the cessation of BCG therapy, detected using a multiplex PCR method as having been caused by BCG. Our report illustrates the efficacy of this method to detect the responsible microbe that is thought to be transmitted from the instillated BCG strain.