Mutational screening of APP gene in patients with early-onset Alzheimer disease utilizing mismatched PCR-RFLP

To elucidate the frequency of mutations of the β/A4 amyloid protein precursor (APP) gene in early-onset Alzheimer disease, we designed a mismatched PCR-RFLP that can identify all kinds of missense mutations at codon 717 in addition to the seven kinds of known mutations at exon 17. When we screened mutations at exon 17 utilizing this method and the double missense mutations at exon 16 of the APP gene by PCR-RFLP, no cases revealed mutations of the APP gene among 13 familial and 54 sporadic cases, except one family (OS-1) that had previously been reported and used as a positive control of APP717(Val → Ile). Our results support the hypothesis that mutations in the APP gene are not major causes in early-onset Alzheimer disease.     

Results of animal experiments using an undulation pump total artificial heart: analysis of 10 day and 19 day survival

An undulation pump is a special rotary blood pump in which rotation of a brushless DC motor is transformed to an undulating motion by a disc in the pump housing attached by means of a special link mechanism. In the blood pump, a closed line between the disc and housing moves from the inlet to the outlet by this undulating disc motion, which sucks and pushes the blood from the inlet to the outlet. Because the same phenomena occurs at both sides of the disc, a continuous flow is obtained when the motor rotational speed is constant. The pump flow pattern can be easily changed from continuous flow to pulsatile flow by controlling the motor drive current pattern. A seal membrane made of segmented polyurethane protects the blood from invading the link mechanism as well as the motor. UPTAH is fabricated with two undulation pumps and two brushless DC motors. Its size is 75 mm in diameter and 80 mm long, and it has one of the great advantage of no compliance chamber required in the system. UPTAHs were implanted under cardiopulmonary bypass (CPB) into the chest cavities of 16 goats, each weighing between 41 and 72 kg. No anticoagulant and antiplatelet agent was used after the surgery. The left atrial pressure was automatically controlled to prevent its elevation and sucking of the atrial wall into the atrial cuff. The following results were obtained: (1) UPTAHs fit well into all the goats; (2) the longest survival was 19.8 days, the cause of death was bleeding from the aortic anastomosis; (3) No thrombus was observed in the blood pump despite no anticoagulant use. Hemolysis depended upon the length of CPB during surgery. When CPB time was within 2 hours, hemolysis level returned to baseline within a few days of the surgery. UPTAH is a promising implantable TAH, because of its small size and easy controllability.     

Renal primitive neuroectodermal tumor: An immunohistochemical and cytogenetic analysis

The cytogenetic and morphologic characteristics of a case with a primitive neuroectodermal tumor (PNET) arising from the left kidney in a 22 year old man are presented. The patient was detected as having a left renal mass with a tumor embolus In the inferior vena cava and multiple pulmonary metastases. A radical nephrectomy with tumor embolectomy of the Inferior vena cava, along with a resection of the pulmonary nodules were performed. Histologic examination revealed a dense proliferation of small round cells with many Homer-Wright type rosettes and perlvascular pseudo-rosettes. Immunohlstochemically, the tumor cells stained strongly positive for HBA71(p30/32^{M IC2}), a surface glycopro-teln specific to PNET and Ewlng's sarcoma. In addition, the tumor cells expressed several neural markers (neuron specific enolase, neurofilament, synaptophysin, and Leu-7) and vimentin, while the epithelial, muscular, and lymphocytlc markers were negative in the tumor cells. Cytogenetic analysis of cultured tumor cells showed a reciprocal translocation t(11; 22)(q24; q12) that is considered to be specific to PNET and Ewing's sarcoma. In conclusion, this case suggested that a karyotyping analysis is a useful diagnostic tool for renal PNET, and it may therefore be utilized to help distinguish between difficult cases of small round cell tumors and Wilms' tumor of the kidney.     

Case report of de novo dup(18p)/del(18q) and r(18) mosaicism

This is a report of a 27-year-old woman with an unusual de novo chromosomal abnormality. Mosaicism was identified in peripheral blood cells examined by standard G-bands by trypsin using Giemsa (GTG) analysis and fluorescence in situ hybridization (FISH) analysis with chromosome-18 region-specific probes, 46,XX,del(18)(pter → q21.33:)[41], 46,XX,r(18)(::p11.21 → q21.33::)[8], and 46,XX,der(18)(pter → q21.33::p11.21 → pter)[1]. On the other hand, the karyotype of periodontal ligament fibroblasts was nonmosaic, 46,XX, der(18)(pter → q21.33::p11.21 → pter)[50]. All cell lines appeared to be missing a portion of 18q (q21.33 → qter). The pattern of the dup(18p)/del(18q) in the rod configuration raises the possibility of an inversion in chromosome 18 in one of the parents. However, no chromosomal anomaly was detected in either parent. The most probable explanation is that de novo rod and ring configurations arose simultaneously from an intrachromosomal exchange. The unique phenotype of this patient, which included primary hypothyroidism and primary hypogonadism, is discussed in relation to her karyotype.     

Colchicine-induced Inhibition of Fat Globule Development in Hepatocytes of Rats Injected with Ethionine

To examine the effect of colchicine on ethionine induced fatty liver, adult female rats were starved overnight and then injected i.p. with 1 g kg ethionine at 11th hour of fasting; then a half of the rats were also injected i.p. with 2.5 mg kg colchicine twice at 3 and 6 h after the single administration of ethionine. Similarly, fasted control rats were injected i.p. with vehicle alone at the above times. All of the rats were sacrificed after a 20 h fast, and the hepatocytes in periportal areas were observed ultra-structurally. In addition, total lipids in the liver tissue were extracted and determined biochemically. Although similar significant increases of triglyceride were observed in the liver tissue of all ethionine-injected rats, the hapatocytes in the group treated with both chemicals had fewer cytoplasmic fat globules (CFG) than those in the group treated with ethionine only. On the other hand, the diameters of markedly increased membrane-bound lipid particles (MLP) in the double treated group were distributed mainly in the range 0.2–0.4 μm, compared with those (0.1-0.2 μm) in the other groups. These findings indicate that colchicine inhibits the development of CFG in ethionine injured hapatocytes. Acta Pathol Jpn 39: 281∼288, 1989.     

Synthesis of block copolymers from 1-chloro-1-octyne and other acetylenes through their sequential living polymerization by MoOCl4?n-Bu4Sn?EtOH

Block copolymerizations of 1-chloro-1-octyne (1-ClO) with several substituted acetylenes were examined by means of living polymerization. o-(Trifluoromethyl)phenylacetylene (o-CF₃PA), o-(trimethylsilyl)phenylacetylene (o-Me₃SiPA), 1-chloro-2-phenylacetylene (1-ClPA), p-butyl-o,o,m,m-tetrafluorophenylacetylene (p-BuF₄PA), and tert-butylacetylene (t-BuA) were used as comonomers, and the MoOCl₄-n-Bu₄Sn-EtOH (mole ratio 1:1:1) catalyst, which is known to effect living polymerization of substituted acetylenes, was employed. When o-CF₃PA and 1-CIPA were the comonomers in combination with 1-CIO, block copolymers were exclusively obtained in both orders of monomer addition. In the cases of o-Me₃SiPA and p-BuF₄PA as comonomers, the copolymerizations initiated from 1-CIO produced block copolymers selectively, whereas the homopolymers of o-Me₃SiPA and p-BuF₄PA also formed if the order of monomer addition was reversed. The pair of 1-CIO and t-BuA did not selectively yield block copolymers irrespective of the order of monomer addition. Thus, block copolymerization occurred between 1-CIO and monomers that show high “livingness” and close reactivities.     

Expression of the Tn antigen on erythroid cells from a patient with Tn syndrome

Summary In order to examine expression of the Tn antigen on erythroid cells from a patient with Tn syndrome, we applied a selective two phase liquid culture system for human erythroid progenitors in peripheral blood. The cells were analyzed with flow cytometry employing an anti-Tn antibody and a lectin ofVicia villosa which recognizes only the Tn determinant. In the second phase, the Tn antigen was expressed on the cultured cells from the patient on day 3 and Tn-positive cells reached 62.7% on day 9. On the other hand, Tn-positive cells were not detected in the volunteer's cultured cells. When the patient's cells were co-cultured with the cells from a healthy voluteer, the percentage of Tn-positive cells was much lower than the expected value, suggesting that the normal cells suppressed the expression of Tn antigen on the patient's cells.     

LIVER CHANGES IN HEPATOCEREBRAL DISEASE*

An attempt of pathogenetic analysis of Wilson's disease and Inose's disease, the two most important varieties of hepatocerebral diseases, was made from the facette of the hepatic pathology. The liver changes of a total of 43 human autopsy cases(19 of Wilson's and 24 of Inose's disease) were reevaluated. Concurrently an attempt of experimental production in animals of hepatic lesions compatible with those of Wilson's disease was made. Inose's special type of hepatocerebral disease is divided into two subgroups, a shunt form and a nutritional form. Cerebral damage in the shunt form is attributed to the portal-systemic collaterals, while the clinicopathological survey of the cases with the nutritional form indicates that functional disturbances, related unbalanced diet or endocrine abnormality, seem to cause brain damage. In Wilson's disease, three stages are noted from the analysis of the liver changes; fibrosis or monolobular cirrhosis in the first stage, submassive necrosis or active postnecrotic cirrhosis in the second stage and multilobular cirrhosis or arrested postnecrotic cirrhosis in the third stage. Experimentally, monolobular cirrhosis could be produced in animals by copper administration alone, whereas postnecrotic cirrhosis was first reproduced by a combination of copper administration and low protein diet, anti-kidney serum, or copper nephrosis. It is suggested that for the development of postnecrotic cirrhosis in the second and third stages of Wilson's disease an additional factor other than copper is needed, and that this second factor may be the deficiency of SH- containing aminoacids.