Functional tyrosine kinase inhibitor profiling: a generally applicable method points to a novel role of platelet-derived growth factor receptor-beta in tuberous sclerosis

A ubiquitous herpesvirus that establishes life-long infection, the Epstein-Barr virus (EBV) has yielded little insight into how a single agent in general accord with its host can produce diverse pathologies ranging from oral hairy leukoplakia to nasopharyngeal carcinoma, from infectious mononucleosis to Hodgkin's disease (HD) and Burkitt's lymphoma. Its pathogenesis is further confounded by the less than total association of virus with histologically similar tumors. In other viral systems, defective (interfering) viral genomes are known to modulate outcome of infection, with either ameliorating or intensifying effects on disease processes initiated by prototype strains. To ascertain whether defective EBV genomes are present in HD, we examined paraffin-embedded tissue from 56 HD cases whose EBV status was first determined by cytohybridization for nonpolyadenylated EBV RNAs (EBERs). Using both standard polymerase chain reaction (PCR) and PCR in situ hybridization, we successfully amplified sequences that span abnormally juxtaposed BamHI W and Z fragments characteristic of defective heterogeneous (het) EBV DNA from 10 of 32 (31%) EBER-positive tumors. Of 24 EBER-negative HD, 8 yielded PCR products indicating presence of het EBV DNA. Two of these contained defective EBV in the apparent absence of the prototype virus. Of the 42 tumors analyzed for defective EBV by both PCR techniques, there was concordance of results in 38 (90%). Detection of defective EBV genomes with the potential to disrupt viral gene regulation suggests one mechanism for pathogenic diversity that may also account for loss of prototypic EBV from individual tumor cells.     

Electron microscopic study of the prenatal development of the thoracic aorta in the rat

Prenatal development of the thoracic aorta of the rat during the period ranging from gestational days 12 to 21 was examined by transmission electron microscopic and morphometric studies. The process of wall formation occurred in four major phases. At phase I (gestational day 12), the dorsal aorta consists of an endothelium and loosely surrounding mesenchymal cells. Collagen fibrils and fine filamentous materials are sparsely present in the intercellular space. At phase II (days 13 to 16), the mesenchymal cells begin to differentiate to myoblasts, which have small clusters of myofilaments with dense bodies, rough endoplasmic reticulum, and a discontinuous basal lamina. The differentiating cells form a few compact cell layers around the endothelium. Elastic fibers first occur sparsely in juxtacellular spaces at days 13–14. The thickness of the aorta increases rapidly from 1–3 layers of cells at day 13 to 5–8 layers at day 17, leading to a maximum of 5–9 cell layers at day 20. The differentiation of myoblasts and elastogenesis are initiated in the inner layers, and later progress toward the outer layer of the aortic wall. At phase III (days 17 to 19), the myoblasts continue to develop into typical smooth muscle cells, and elastic fibers rapidly increase in both size and number. At phase IV (day 20 and later), smooth muscle cells have well-developed myofilaments in the cell periphery, and rough endoplasmic reticulum and other organelles tend to accumulate in the apical portion of the cytoplasm. Elastic laminae appear in a few inner layers of the aortic wall.     

BRONCHIOLO-ALVEOLAR ADENOCARCINOMA WITH MULTIPLE CYSTS

An autopsy case of bronchiolo-alveolar adenocarcinoma in the lung is reported. The patient is a 70-year-old male who complained of severe cough with 500–600 ml watery sputum a day, loss of weight, and general fatigue. Autopsy revealed numerous whitish tumors in various sizes with multiple cysts in both lungs, with no metastasis being found in any other organs. Histological findings identified the tumor as a bronchiolo-alveolar adenocarcinoma originating from the lungs. Electron-microscopic findings showed that the tumor cells were covered by prominent microvilli, and contained abundant irregulary-shaped cytoplasmic vacuoles suggestive of mucin.     

Four cases of wheat-dependent exercise-induced anaphylaxis with negative gluten cap-rast score]

BACKGROUND: Either omega-5 gliadin or high molecular weight glutenin is known to be a major allergen in wheat-dependent exercise-induced anaphylaxis (WDEIA). It is generally considered that gluten specific IgE score is more reliable than that of wheat specific IgE score for the diagnosis of WDEIA. Our aim was to verify the significance of gluten specific IgE in the diagnosis of WDEIA. METHODS: We evaluated the result of gluten CAP-RAST score and omega-5 gliadin specific IgE score on four WDEIA patients who visited our hospital during the years 2004 and 2005, whose diagnosis were onfirmed by prick tests, immunoblot tests and provocation tests. RESULTS: Contrary to our expectations, all four patients showed negative gluten CAP-RAST scores, however all patient's omega-5 gliadin specific IgE scores were positive. CONCLUSION: Our results suggest that gluten specific CAP-RAST score is unreliable in the diagnosis of WDEIA. On the other hand omega-5 gliadin specific IgE score is possibly a better candidate as a diagnostic tool for WDEIA.     

Koebner phenomenon in lupus erythematosus with special consideration of clinical findings

The isomorphic response of Koebner can be observed not only in psoriasis, but also in other diseases, such as lichen planus and some systemic diseases including LE (lupus erythematosus) or sarcoidosis. Several clinical findings in LE skin were presented and discussed in this review. The mutually-interactive-, negative-, and internal-Koebner phenomena were introduced and discussed with some speculative views. Many forms of environmental stress on the skin were reported as provocating factors of the Koebner phenomenon, including trauma, scratching, UV-exposure, and various types of dermatitis. Clinical observations of the nature, localization, and movement of lesions should be carefully made. The pathophysiology of the Koebner phenomenon may be classified into two steps. A first non-specific inflammatory step and a second disease-specific step. The inflammatory products released from the first step would be targeted in the second step. In the first step, there could be many substances including cytokines, stress proteins, adhesion molecules, or autoantigens translocated from intra-cellular areas. In the second step after latent periods, there may be disease-specific reactions, including ones by T-cells, B-cells, autoantibodies, and immune deposits, under the restriction of genetic backgrounds. The Koebner phenomenon may prove useful in understanding the pathophysiology of diseases of unknown origin.     

JCCLS phlebotomy standards

In 2004, the Japanese Committee of Clinical Laboratory Standards (JCCLS) published a standard phlebotomy guideline, which not only ensures the safety of the patients and phlebotomists but is adopted to the healthcare setting in Japan. This phlebotomy standard is also essential for the standardization of clinical laboratory tests. This guideline was completed on the basis of current phlebotomy procedures widely in use in Japan using phlebotomy standards in the USA as references, while reconsidering their scientific reasoning as far as possible. At the same time, factors such as practicality and cost benefit were taken into account. The content of the guideline includes necessary facilities and equipment, a step by step safe but practical venipuncture procedure, an explanation of the individual steps, and other supplementary information such as alternative methods. The first edition, published as tentative guideline, is planned to be revised after a set period of time based on the comments and suggestions from a wide range of people concerned, so that it can be published as an approved guideline.     

Mechanisms responsible for delayed and immediate hemolytic transfusion reactions in a patient with anti-E + Jk(b)+ Di(b) and anti-HLA alloantibodies

Immediate hemolytic transfusion reactions (IHTR) occurred in the course of delayed hemolytic transfusion reactions (DHTR). An 84-year-old man had received a blood transfusion 20 years ago. Progressive anemia developed, because of continuous bleeding from a bladder tumor. He was transfused with concentrated red blood cells (CRC) which were Rh-E antigen negative, because he had anti-E antibodies (day 0). He received CRC on day 3, and underwent resection of bladder tumor on day 6. Although crossmatch-compatible CRCs were prepared for the operation, those were not required and were kept in a refrigerator in the ward. On day 9, when a CRC kept in the ward was transfused, he suddenly had a IHTR. In order to analyze a mechanism of IHTR, the anti-Jk(b) and anti-Di(b) antibodies, anti-HLA antibodies and the concentrations of inflammatory cytokines were measured in serum samples. The anti-Jk(b) and anti-Di(b) antibodies increased prior to IHTR experienced on day 9. The concentrations of IL-6 and IL-1beta increased from day 2, while the concentration of IL-8 increased from day 7. The anti-HLA class I antibody could be detected 2 days before IHTR. Thus, the anti-Jk(b) and anti-Di(b) antibodies induced the production of inflammatory cytokines and symptoms of DHTR and IHTR. The anti-HLA class I antibody could be produced in spite of using the filer for removing leukocytes, and may take part in the induction of IHTR. Further, blood products should be transfused soon after completing a crossmatch test in patients with anti-RBC alloantibodies.     

Distribution and frequencies of PDS (SLC26A4) mutations in Pendred syndrome and nonsyndromic hearing loss associated with enlarged vestibular aqueduct: a unique spectrum of mutations in Japanese

Molecular diagnosis makes a substantial contribution to precise diagnosis, subclassification, prognosis, and selection of therapy. Mutations in the PDS (SLC26A4) gene are known to be responsible for both Pendred syndrome and nonsyndromic hearing loss associated with enlarged vestibular aqueduct, and the molecular confirmation of the PDS gene has become important in the diagnosis of these conditions. In the present study, PDS mutation analysis confirmed that PDS mutations were present and significantly responsible in 90% of Pendred families, and in 78.1% of families with nonsyndromic hearing loss associated with enlarged vestibular aqueduct. Furthermore, variable phenotypic expression by the same combination of mutations indicated that these two conditions are part of a continuous category of disease. Interestingly, the PDS mutation spectrum in Japanese, including the seven novel mutations revealed by this study, is very different from that found in Caucasians. Of the novel mutations detected, 53% were the H723R mutation, suggesting a possible founder effect. Ethnic background is therefore presumably important and should be noted when genetic testing is being performed. The PDS gene mutation spectrum in Japanese may be representative of those in Eastern Asian populations and its elucidation is expected to facilitate the molecular diagnosis of a variety of diseases.     

Bone resorption versus estrogen loss following oophorectomy and menopause

OBJECTIVES: To investigate if menopause and oophorectomy may represent different risk factors for bone resorption/loss. METHODS: The urinary levels of pyridinoline (Pyr) and deoxypyridinoline (D-pyr), the serum levels of type I carboxy-terminal pyridinoline cross-linked telopeptide (ICTP), and lumbar bone mineral density (BMD), were compared in 80 Japanese women after menopause or oophorectomy. These women were divided into four groups of 20 women each as follows: early postmenopausal stage (early physiologic menopause < 3 years before study entry); late postmenopausal stage (physiologic menopause > or = 3 years before study entry); early postoophorectomy stage (oophorectomy < or = 03 years before study entry); or late oophorectomy stage (oophorectomy > 3 years before study entry). RESULTS: Lumbar BMD was significantly lower in the late groups compared to their respective early groups and was lowest in the late postoophorectomy group. The ratio of D-pyr/creatinine (Cr) was not significantly different among the four groups. The ratio of Pyr/Cr was significantly higher in the early postoophorectomy subjects compared with either late group. The serum level of ICTP was significantly higher in the early postoophorectomy group compared to all other groups. CONCLUSIONS: These findings suggest that serum ICTP may be useful in detecting changes in bone resorption after oophorectomy and that women are at greater risk for bone resorption after oophorectomy than after physiologic menopause, although this difference appears to diminish with time.