Real-time PCR method for identification of Asian populations in forensic casework.

The priorities for mass disaster victim identification are rapid investigation turn-around time and low cost. We describe a DNA typing procedure to selectively identify members of Asian populations by a real-time PCR method using polymorphisms of the alpha2 chain of the type I collagen gene (COL1A2) and mitochondrial DNA (mtDNA). Among the 50 members of the Asian population included in the present study, 37 harbored a deleted allele in intron 33 of COL1A2 (26822-26823del) or the 10400C>T substitution mutation in mtDNA to give a probability of 0.740 for these SNPs in the Asian population.

Genetic polymorphism in collagen type 1 alpha 2 intron 33 can be used for the initial screening of East Asians quickly and easily by a heteroduplex analysis.

To save time and simplify the matching genetic information of bereaved family with the collected samples for identification of individuals in a mass-scale disaster and terrorism using molecular biological methods, we propose a method to initially screen samples according to different populations. We found a unique deletion (26822-3) in an allele that 62.0% of East Asian individuals had in intron 33 of type I collagen alpha 2 (COL1A2), while most Europeans and Africans did not have it. By making a heteroduplex complex, a visual discrimination was possible using 10% PAGE. A PCR and heteroduplex analysis could identify genetic polymorphism of COL1A2 intron 33 both clearly and accurately, even from decomposed samples or very small amounts of samples within a very short time period (approximately within 150min). As a result, genetic polymorphism of COL1A2 intron 33 is thus considered to be useful to quickly and easily screen East Asians from vast number of samples using a heteroduplex analysis. Consequently, we consider that an initial screening method for East Asians which use a heteroduplex analysis of genetic polymorphism of COL1A2 intron 33 can smoothly and effectively perform various identifications among East Asians when supplies are logistically poor after a mass-scale disaster or terrorism.

[Medico-legal autopsy case of an infant suffering anaphylactic shock during dental treatment. Potential hazards in the use of a rubber-dam-sheet for infants].

We report a medico-legal autopsy of a 4-year-old girl without any history of allergies, who died following anaphylactic shock due to local anesthesia with lidocaine during dental treatment. The shock symptoms were overlooked due to a rubber-dam-sheet covering her face. The autopsy revealed laryngeal edema, secretory fluid filling the trachea, severe pulmonary congestion and edema. An immunohistochemical investigation showed numerous mast cells releasing histamine in lungs, and the fluids in the pulmonary alveoli and veins also showed positive staining. In addition, the plasma concentration of histamine in the heart blood showed a high value. From these findings, the cause of death was determined to be respiratory failure due to anaphylactic shock induced by lidocaine. In this case, her death was considered to be due to medical malpractice; adequate life support had not been performed, because anaphylactic shock was overlooked until the rubber-dam-sheet removed. In addition, due to two previous autopsy case reports that described infantile fatalities involved with the misusage of a rubber-dam-sheet for the patients under local anesthesia, we therefore recommend immediate improvements in rubber-dam-sheets and/or better applications of them in pediatric dentistry.

[Tuberculous lymphadenitis: a clinical study of 23 cases].

INTRODUCTION: Tuberculous lymphadenitis is a relatively rare disease in adults. In the absence of pulmonary tuberculosis, tuberculous lymphadenitis is very difficult to differentiate from other diseases. We described our experiences of patients with tuberculous lymphadenitis. RESULTS: We diagnosed 23 patients with tuberculous lymphadenitis out of 207 patients with tuberculosis. Their ages ranged from 18 to 99 years (mean, 45.7 years), and the male-to-female ratio was 7:16. The most common complaints were cervical mass and fever. With the exception of two patients, all diagnosed patients had a strong positive skin test to tuberculin. Observing the site of affected lymph nodes, 16 patients had cervical node involvement, 3 patients had axillary node involvement, 7 patients had mediastinal node involvement, 3 patients had hilar node involvement, 3 patients had abdominal node involvement, and 1 patient had inguinal node involvement. Fifteen patients had neither hilar nor mediastinal node involvement. Eleven patients had no tuberculous lesions other than lymphadenitis. Seven patients underwent biopsy of the lymph nodes. Four of these patients had the evidence of acid-fast bacilli. The remaining three patients were also diagnosed histologically. Five patients underwent fine needle aspiration. Two of them had the evidence of acid-fast bacilli. Acid-fast bacilli were detected in 10 out of 16 sputum samples and in 1 out of 2 pleural effusion samples. Five patients were diagnosed clinically by image (Computed tomography etc.) and by therapeutic effect. Eleven cases underwent contrast-enhanced computed tomography (CT) of the lymph nodes. Seven cases showed central low attenuation with peripheral rim enhancement, whereas the other four cases showed homogeneous attenuation. All patients received chemotherapy for a mean duration of 14.5 months (range, 6-30 years) with apparent improvement, but 1 patient relapsed. CONCLUSION: Tuberculous lymphadenitis remains one of important targets for the differential diagnosis of lymphadenopathy. It is essential that a peripheral lymph node biopsy be performed and examined either histologically and/or microbiologically. A tuberculin skin test and contrast-enhanced CT imaging should also be performed.

[A case of miliary tuberculosis complicated with a tuberculous aneurysm of the aorta].

A 68-year-old man was admitted because of fever and weight loss. A chest high-resolution computed tomography (HRCT) scan revealed diffuse micronodular shadows, and an abdominal CT scan showed an aneurysm spreading from the root of the renal artery to the iliac bifurcation. His fever subsided without treatment and his general condition was good. However, histological studies of a transbronchial lung biopsy specimen and bone marrow aspirate clot revealed non-necrotizing epitheloid granulomas, and mycobacterium tuberculosis was cultured from sputum and bronchoalveolar lavage fluids. Military tuberculosis was diagnosed. One month after the initiation of chemotherapy, the abdominal aneurysm enlarged quickly despite the improvement of the thoracic findings, and graft replacement was performed. Histological findings in specimens of the resected aneurysm suggested that tuberculous inflammation of the surrounding lymph nodes had invaded the aortic wall, leading to the aneurysm. It should be borne in mind that symptoms in elderly patients with miliary tuberculosis may be mild, even when serious extrapulmonary lesions are present.

[An elderly patient with advanced non-small-cell lung cancer that responded remarkably to combination chemotherapy of vinorelbine and gemcitabine].

A 73-year-old man was admitted to our hospital because of hemoptysis in November 1999. A chest CT revealed a mass shadow in the right upper lobe and enlarged hilar and pretracheal lymph nodes. Bone scintigraphy showed a bone metastasis in right middle rib. He was treated with vinorelbine (25 mg/m2 days 1,8) and gemcitabine (800 mg/m2 days 1,8) every 3 weeks from December 1999. After 4 courses of chemotherapy, the primary tumor and hilar lymph node were remarkably reduced in size, and his hemoptysis disappeared and body weight increased. It is suggested that combination chemotherapy with vinorelbine and gemcitabine is effective for elderly patients with non-small-cell lung cancer with good performance status.