A novel point mutation affecting the tyrosine kinase domain of the TRKA gene in a family with congenital insensitivity to pain with anhidrosis.

A nerve growth factor receptor encoded by the TRKA gene plays an important part in the formation of autonomic neurons and small sensory neurons in dorsal root ganglia and in signal transduction through its intracytoplasmic tyrosine kinase domain. Recently, three mutations in the tyrosine kinase domain of TRKA have been reported in patients with congenital insensitivity to pain with anhidrosis, which is an autosomal recessive disorder characterized by recurrent fever due to absence of sweating, no reaction to noxious stimuli, self-mutilating behavior, and mental retardation. We examined the TRKA gene in five generations of a large Japanese family with many consanguineous marriages who live in a small remote island of the southern part of Japan. We found a novel point mutation at nucleotide 1825 (A-->G transition) resulting in Met-581-Val in the tyrosine kinase domain. Two of the three affected patients were homozygous for this mutation; however, the third affected patient was heterozygous. Further analysis revealed that the third patient was a compound heterozygote with the Met-581-Val mutation in one allele and with a single base C deletion mutation at nucleotide 1726 in exon 14 in the other allele, resulting in a frameshift and premature termination codon.

Virilizing adrenal adenoma stimulated by dexamethasone in a middle-aged woman.

In a middle-aged woman with virilizing adenoma, 2 mg dexamethasone increased urinary excretion of 17-ketosteroids (17-KS) and 17-hydroxycorticosteroids, whereas 8 mg dexamethasone increased urinary excretion only of 17-KS. With discontinuation of dexamethasone, 17-KS excretion returned to the predexamethasone level. Dexamethasone depressed the basal level of cAMP synthesis and basal testosterone production by the normal adrenal tissue in vitro. Dexamethasone also depressed the increase of cAMP produced by ACTH in the normal tissue. In contrast, dexamethasone increased basal cAMP synthesis and stimulated testosterone secretion in the tumor tissue. ACTH and dexamethasone were additive in their effects on cAMP and testosterone in the tumor tissue. It is suggested that dexamethasone acted directly on the adrenal tumor to stimulate steroid secretion in this patients.

Ultrastructural immunocytochemical studies of blood group substances in human eccrine glands.

We investigated the ultrastructure of blood group antigens A, B, and H in human eccrine glands by means of the immunogold labeling technique. Blood group antigens A, B, and H were found in the Golgi apparatus, secretory granules, and over the apical and basolateral cell membranes of dark cells of eccrine glands depending on the blood group phenotype of the donors. Both A and B antigens were found in the dark cells of AB donors. The labeling pattern of the Golgi stacks seemed to have a polarity whereby the anti-blood group A antibody labeled all the stacks, whereas anti-blood groups B and H bound to the trans side of the Golgi complex. These observations suggest that the blood group substances are secreted into the lumen after being processed through the Golgi apparatus and the immature and mature granules in the dark cells of human eccrine glands.

Failure to detect paramyxovirus RNA in the skin of connective tissue disease.

Etiology of connective tissue disease is unknown. The association of infectious agents has been suspected serologically, ultrastructurally and recently by means of molecular biological techniques. We extracted RNA from lesions of 25 discoid lupus erythematosus, 9 systemic LE, and 3 systemic sclerosis biopsies, and reverse transcription-polymerase chain reaction was performed for all paramyxoviruses known to infect human beings. None of the samples yielded positive signal for any of the viruses. The existence of paramyxovirus in the skin of connective tissue disease is very unlikely.

Analyses of the transglutaminase 1 gene mutation and ultrastructural characteristics in a Japanese patient with lamellar ichthyosis.

We described a Japanese female with lamellar ichthyosis whose transglutaminase 1 gene (TGM1 gene) was mutated. DNA sequence analysis revealed that the patient had a homozygous mutation, i.e. a point mutation from G to A at nucleotide 1494 resulting in the substitution of glycine for arginine at codon 143. Her mother was heterozygous for this mutation. In situ transglutaminase assay in the patient's skin showed loss of enzyme activity. Ultrastructural examination revealed incomplete formation of cornified cell envelopes and electron-dense materials adjacent to plasma membranes. These results suggest that defective transglutaminase activity caused by homozygous TGM1 gene mutation (G143R) results in disruption of cornified envelope assembly and the clinical phenotype of lamellar ichthyosis.

Subtyping hepatitis B virus DNA in free or integrated forms by amplification of the S-gene sequences by the polymerase chain reaction and single-track sequencing for adenine.

The S-gene fragments of hepatitis B virus (HBV) DNA in serum, or integrated in chromosomes of human hepatoma cells (PLC/PRF/5), were amplified by the polymerase chain reaction, cloned into an M13 phage vector, and then sequenced only for adenine. The subtype determinant d or y was established by the presence or absence of adenine as nucleotide 365, and w or r by that of nucleotide 479 in the S gene. The results were identical with those obtained by enzyme immunoassay with monoclonal antibodies. A high sensitivity for the detection of HBV DNA, amplified by the polymerase chain reaction, allowed subtyping of HBV in sera containing HBsAg in concentrations too low to be subtyped by immunological methods. Furthermore, subtyping at the nucleotide level can be applied to tissues containing HBV DNA sequences in integrated forms, such as hepatocellular carcinomas, stored frozen or in formalin.

Chronic lupoid leishmaniasis. Evaluation by polymerase chain reaction.

BACKGROUND: The cutaneous lesions in chronic lupoid leishmaniasis resemble those of lupus vulgaris, both clinically and histologically. The differential diagnosis is difficult and may depend on the detection of a few Leishmania amastigotes in the histologic sections, the growth of the promastigotes in cultures, or the identification of amastigotes by other techniques. Polymerase chain reaction was used to detect Leishmania amastigote DNA in tissue samples obtained from 65 patients with chronic lupoid leismaniasis, and the results were confirmed by Southern blot analysis. OBSERVATIONS: The histologic findings of a predominantly epithelioid cell granuloma surrounded by lymphocytic infiltrate in chronic lupoid leishmaniasis are very similar to those observed in lupus vulgaris. Extensive histologic examination of the sections in this series revealed occasional macrophages containing a few amastigotes in only 12 cases. Cultures in NNN medium yielded Leishmania promastigotes in 20 cases. Polymerase chain reaction studies using a Leishmania-specific primer identified Leishmania DNA in 30 of 63 cases, and those using a Mycobacterium tuberculosis primer were found to be negative for mycobacteria in 47 cases tested, including 11 cases with a positive tuberculin skin reaction. CONCLUSIONS: The histologic findings in chronic lupoid leishmaniasis resemble those of lupus vulgaris. Polymerase chain reaction studies were useful in identifying amastigotes in 30 (47.6%) of 63 cases. This study confirms the presence of DNA molecules of Leishmania amastigotes in samples of formalin-fixed, paraffin-embedded granulomatous tissue obtained from patients with chronic lupoid leishmaniasis.

Histopathologic and ultrastructural studies of angiokeratoma corporis diffusum in Kanzaki disease.

A novel metabolic disease, angiokeratoma corporis diffusum (Kanzaki), was the subject of an extensive histopathologic and ultrastructural study. Findings included dilated lymph and blood vessels in the upper dermis with an orthokeratortic, thickened, horny layer in well developed angiokeratoma. In the early papules, a few sporadic dyskeratotic keratinocytes were present in the epidermis with or without a thickened horny layer. Vesicular clear vacuolation was clearly observed in the cytoplasm of the secretory portion of the eccrine sweat glands, but none was observed in the vascular endothelial cells with hematoxylin-eosin staining. Using electron microscopy, lysosomal vacuolation was observed in many cell types, including eccrine sweat gland cells, vascular endothelial cells, dermal fibroblasts, dermal neural cells, lymphocytes of peripheral blood in the skin, and glomerular endothelial cells, but none was noted in the epithelial cells of the kidney. Widely dilated vacuoles were found to contain only a small amount of fuzzy filamentous material in the vascular endothelial cells, filamentous or electron-dense granular substances in fibroblasts, and electron-dense, lamellated or homogeneous structures in eccrine sweat gland cells and in neural cells. Ultracytochemical examination revealed glycoconjugates in dilated lysosomes. Characteristics of Kanzaki Disease were shown to differ from those of Fabry disease or any other lysosomal storage disease.

Anaplastic large cell lymphoma, so-called Ki-1 lymphoma: a report of a case of long course.

A 55-year-old female with a complaint of tumors on the right lower extremity was reported. The condition was diagnosed as anaplastic large cell lymphoma, so-called Ki-1 lymphoma, by its histological and immunohistochemical features. The clonal proliferation of the infiltrating cells of the skin lesions was confirmed by the analysis of T cell receptor gene rearrangement. The lesions have repeatedly occurred on the right lower extremity for more than ten years. In this report, we also discuss the prognosis of anaplastic large cell lymphoma with or without skin lesions.

A case of chromomycosis with tumor-like growth.

A 66-year-old woman who lived on Tokunoshima Island, a small and remote southern island of the Japanese archipelago, had suffered from chromomycosis for more than 30 years and presented with a tumor-like growth on the posterior crural region of his right leg. Fonsecaea pedrosoi was identified as the pathogen from its growth pattern and micromorphological characteristics. The patient was successfully treated with 5-fluorocytosine, itoraconazole, and topical thermotherapy.

[Epidemiological study for comparative biological profiles on MRSA strains isolated in 1992 vs. 1993].

The trend of epidemiological study against MRSA strains which were isolated in 1992 and in 1993 was investigated. Number of stains tested yearly consisted of 30 isolates that were considered to play pathogenic roles for inpatients in clinical departments at our institute. In comparing with biological studies on MRSA strains and the epidemiological surveillance of the background of the isolation, the data summarizes as followings; 1) No. of MRSA strains which were producible for TSST increased from 24/30, 80% up to 30/30, 100%. 2) No. of enterotoxin type harbouring biotype of B/C increased 0/30, 0% up to 12/30, 40%. 3) No. of type of plasmid DNA profile increased in varying from 3 types (A, B, C) to 8 types (A-H). 4) The in vitro activity of antimicrobials, as such MINO, GM, IPM, CMZ was less potent than that of the prior year, and even for VCM, ABK, the activity proved less potent in 1-2 tubes in MIC90. 5) No. significant hospital acquired infection was detected between the inpatients, with MRSA infection and isolates from plasmid DNA profiles. 6) Since the ratio of the coincidence of plasmid DNA profiles of MRSA was only in 4 patients out of 27, 14.9 &, nosocomial infections with MRSA brought to patients have not only been considered by medical, paramedical staff, but that the infection may be caused by broad contamination at the institute.

[Autopsy case of adenoid cystic carcinoma of the trachea: endobronchial treatment improves quality of life].

A 67-year-old man presented with dyspnea on exertion. Bronchoscopic examination revealed a tumor arising from the middle portion of the trachea and extending to the right main bronchus. The pathological diagnosis was adenoid cystic carcinoma. Radiotherapy and subsequent endobronchial electrocautery were performed, and elicited a partial response. In the clinical course. Dumon and Ultraflex stents were placed in the trachea asynchronically. Brachytherapy and esophageal stent placement were also performed for tumor control in the trachea and esophagus. Autopsy revealed that the tumor had invaded the trachea and esophagus, and bacterial mediastinitis was also demonstrated. Because the tumor was successfully controlled during the following 4 years and 9 months, we concluded that endobronchial therapy such as stent placement or electrocautery is useful for maintaining good quality of life.

Selenium deficiency: report of a case.

We report an 18-month-old Japanese boy with selenium deficiency. He had dry skin with irregularly shaped, erythematous changes on the cheeks, groin, hip, and extremities, erosions on the external urethral and anal orifices, and sparse, short, thin, light-coloured hair. He had received parenteral nutrition for 5 months because of juvenile polyposis. At presentation, his serum selenium level was less than 2.0 microg/dL (normal range, 10.6-17.4 microg/dL). His skin lesions responded well to supplementary treatment with sodium selenite. His skin symptoms were similar to those attributable to a deficiency of zinc which, like selenium, is an essential trace element. According to the literature, selenium deficiency is responsible for cardiomyopathy, which was diagnosed in our patient. The clinical similarity to zinc deficiency and the literature yielded important clues for a diagnosis of selenium deficiency in this patient.

The distribution of concanavalin A- and cationized ferritin-binding sites on plasma membrane of human platelet and the changes of these sites in cells responding to adenosine diphosphate.

Investigations were carried out to study the distribution of the concanavalin A- and cationized ferritin-binding sites on the plasma membrane of human platelets and to ascertain the changes of these sites in cells stimulated with adenosine diphosphate (ADP) with time as well. The concanavalin A-binding sites of the unwashed fixed platelets were sparsely distributed on the plasma membrane at a distance of up to 80 nm from the outer leaflet of the plasma membrane. The washed unfixed platelets, however, showed a dense distribution within a range of 70 nm from the outer leaflet of the plasma membrane. Changes in the distribution of concanavalin A-binding sites on the plasma membrane of platelets stimulated with ADP were characterized by a marked increase in the number of binding sites and by protrusion up to a distance of 150 nm from the outer leaflet of the plasma membrane 1 minute after the reaction had occurred. The concanavalin-A binding sites may be semicryptic in view of the fact that they were exposed by washing or protruded as a result of the stimulation with ADP. The cationized ferritin binding sites were uniformly distributed with high density on the plasma membrane of the unwashed fixed platelets. In washed unfixed platelets, however, they were sparsely distributed with cluster formation. It is suggested that the glutaraldehyde fixation itself has an effect on the binding of the cationized ferritin particles on the plasma membrane of platelets. The various changes in the concanavalin A-binding sites appearing 1 minute after the reaction with ADP may represent morphological evidence indicating that the platelets have acquired adhesiveness.

Type III glycogenosis with deposition of urate and amyloid.

A case of a 44-year-old man with hepatic form of glycogenosis was presented. The patient had abdominal distension and muscular weakness. The glucose tolerance test showed a diabetic pattern, though he had hypoglycemia in fasting state. The fructose tolerance test showed an ability of conversion from fructose to glucose. The double glucagon test showed no rise of blood glucose in fasting state but a rise 2 hours after meal. These symptoms and laboratory data supported the clinical diagnosis of type III glycogenosis. At autopsy, glycogen was markedly deposited in the liver, and slightly in the kidneys and heart. The glycogen pooled in the hepatic cells histochemically showed a normal reaction to several glycogen stainings. Electron microscopy by using Thiéry's method revealed that the pooled glycogen particles were clearly arranged as rosettes measuring 1,000A in largest diameter composed of clustered monoparticulates. There were marked hyalinization of the islets of Langerhans containing amyloid. As to its pathogenesis, this change can be interpreted as a morphological expression of the hypofunction of beta-cells ascribed to long-standing hypoglycemia.