Cloning and characterization of excitatory amino acid transporters GLT-1 and EAAC1 in canine brain.

Excitatory amino acid transporters play important roles in termination of glutamatergic neurotransmission and protection of neurons from the excitotoxicity of glutamate in the central nervous system. We herein report isolation of cDNA clones of two distinct excitatory amino acid transporters, GLT-1 and EAAC1, from canine brain cortex by PCR-based cloning, and characterization of these transporter subtypes. Canine GLT-1 and EAAC1 exhibited Na+-dependent glutamate transport with high affinities in a Xenopus oocyte expression system. Despite the similarity in transport kinetics and in the predicted primary structures, GLT-1, EAAC1, and the previously identified GLAST showed different sensitivities to several structural analogues of L-glutamate. In addition, transcripts of these transporter subtypes showed distinct regional distribution in the brain in RT-PCR analysis, suggesting that excitatory amino acid transporters have distinct physiological and pathophysiological roles in the brain.

[Comparison of crossing-over between 30-minute drip infusion vs 30-second injection of granisetron for nausea and vomitting with cisplatin].

Recently, Granisetron (KYT) was proved to have a strong effect for cisplatin (CDDP)-induced emesis. We compared the effect of KYT for CDDP-induced emesis between two different administration schedules. Forty micrograms/kg of KYT was administered either by 30-minute drip infusion with 100 ml of saline (Group A) or 30-second injection with 10 ml of saline (Group B). We investigated the therapeutic effect of KYT in both group A and Group B by the crossing-over method. After the patients who had a malignant tumor and were going to receive CDDP (over 50 mg/m2) in two courses were selected, KYT was administered by the method of Group A or Group B in a double-blind comparison. The clinical efficacy was at least "effective" in 70% (7/10) of Group A and Group B. The study treatment was considered "useful" in 80% (8/10) of Group A, 90% (9/10) of Group B, and "safe" in 100% of Group A and B. There was no difference between two groups in this respect. The results showed that the slow intravenous injection of KYT also has an excellent antiemetic effect on CDDP-induced emesis and a high degree of safety.

Defective anion transport and marked spherocytosis with membrane instability caused by hereditary total deficiency of red cell band 3 in cattle due to a nonsense mutation.

We studied bovine subjects that exhibited a moderate uncompensated anemia with hereditary spherocytosis inherited in an autosomal incompletely dominant mode and retarded growth. Based on the results of SDS-PAGE, immunoblotting, and electron microscopic analysis by the freeze fracture method, we show here that the proband red cells lacked the band 3 protein completely. Sequence analysis of the proband band 3 cDNA and genomic DNA showed a C --> T substitution resulting in a nonsense mutation (CGA --> TGA; Arg --> Stop) at the position corresponding to codon 646 in human red cell band 3 cDNA. The proband red cells were deficient in spectrin, ankyrin, actin, and protein 4.2, resulting in a distorted and disrupted membrane skeletal network with decreased density. Therefore, the proband red cell membranes were extremely unstable and showed the loss of surface area in several distinct ways such as invagination, vesiculation, and extrusion of microvesicles, leading to the formation of spherocytes. Total deficiency of band 3 also resulted in defective Cl-/HCO3- exchange, causing mild acidosis with decreases in the HCO3- concentration and total CO2 in the proband blood. Our results demonstrate that band 3 indeed contributes to red cell membrane stability, CO2 transport, and acid-base homeostasis, but is not always essential to the survival of this mammal.

[Primary torsion of the omentum--report of a case and review of the Japanese literature].

A case of primary torsion of the omentum is reported. A 37-year-old male was admitted with the chief complaint of RLQ pain of 3 day's duration on June 10, 1986. Tenderness and Blumberg's sign were found at Mc Burney's point with leucocytosis of 11,500/mm3. His body temperature was not elevated. Operation was carried out under the diagnosis of suspicion of acute appendicitis. Laparotomy revealed a dark-red omental mass of 12 X 5 X 1 cm which had been formed by torsion (7 times, clock-wise) and a little clear ascites. The omental mass was resected, and appendectomy was also performed. The postoperative course was uneventful. Eighteen cases including this case in the Japanese literature were reviewed and reported.

Alveolar echinococcosis of the liver; studies on 60 operated cases.

During the years 1936 through 1978, 60 cases received surgical intervention for alveolar echinococcosis of the liver. The resectability and operative mortality rate were 64.0% (16/25) and 43.8% (7/16) before 1968, but 54.3% (19/35) and zero (0/19) thereafter. Establishment of clinical staging and criteria for justifying radical resection of a given lesion, combined with systematic evaluation of all hepatic vasculatures, contributed to improvement of the result. The long-term prognosis of the disease, unless resected, has been exclusively poor. A mass screening program, which became possible by the development of serologic tests, has covered a population of over 140,000 in the endemic area and been of value in detecting the disease in its early developmental stage. The disease should be recognized even in currently unaffected areas since the cestode has a fairly wide geographic distribution including the United States.

[Variation of serum free amino acids in fulminant hepatitis treated with hepatic assists (author's transl)].

A 10 year old boy, in grade IV hepatic coma, was treated by combination of XAD-4 resin hemoperfusion (HP), activated charcoal HP (Adsorba 300C, Gambro), exchange transfusion by up to 12.0 liters of fresh whole blood, and regular dialysis. Serum free amino acids' values were consecutively assessed during 4 days of treatment. The liver was 490 gr in weight at autopsy and histologic examination revealed cellular necrosis compatible with fulminant hepatitis. Pre-treatment values of alanine, lysine, proline, phenylalanine, arginine, threonine, tyrosine and methionine were increased by 2 to 38 times of normal control, while those of cystine, glutamic acid, serine and glycine were minimally increased up to 1.7 times. Histidine, isoleucine, leucine and valine, on the other hand, were decreased by 20 to 30% and aspartic acid was the lowest at 14% of normal control. The effect of XAD-4 resin HP and exchange transfusion was rather non-specific by decreasing the total amount of amino acids. The molar ratios of branched chain amino acids vs. aromatic amino acids or essential amino were elevated by activated charcoal HP, but, did not reach to normal range.

Operative treatment of alveolar echinococcosis of the liver.

This study is based on the experience with 51 cases of hepatic alveolar echinococcosis underwent operative treatment up to the end of 1976. Hepatic resection was carried out in 28 cases with overall operative mortality of 25 per cent, but no death occurred in the last 10 years period during which 12 cases were subjected to the procedure. This apparent improvement of the result is ascribed to the establishment of the strict criteria for operative intervention, i.e., when less than three segments are involved, the hepatic hilum is not highly involved, and the inferior vena cava is not invaded. Marsupialization is employed when hepatic resection is not indicated and the lesion shows liquefaction. Ten cases underwent the procedure with one operative death. Biliary tract reconstruction was carried out in two cases with hilar involvement, but the prognosis was poor. Eleven cases were only with celiotomy. The follow-up studies indicated that the hepatic resection offers the best hope for cure followed by marsupialization. Unlike unilocular echinococcosis in which a cyst grows expansively, the alveolar echinococcosis should be considered clinically malignant, in that it grows invasively and often shows metastatic lesions. Surgical intervention at its early developmental stage is the only definitive way of the treatment. Only recent advances in diagnostic procedures and development of type specific serological studies made it possible to bring the disease under control.

Experimental bladder substitution using a biodegradable graft of natural tissue.

A controlled canine experimental study has been performed to test the application of a biodegradeable graft to vesical augmentation. The graft, after providing initial bladder enlargement, is progressively reabsorbed while serving as a foundation for the regeneration and healing of the layers of the bladder remnant. Follow-up studies at 1 year revealed no urinary infection, calculi, extravasation of urine, or rejection of the material. Normal bladder capacities were maintained throughout the study and the reconstructed bladders functioned normally. The biodegradable graft was equally successful when applied either to a histologically normal bladder remnant, or to a pathological bladder of intramural fibrosis. These results suggest that this material may be well suited to clinical bladder substitution in humans.

Carcinoma arising in the cyst of the liver--report of three cases--.

A generally recognized concept dictates that surgical intervention for non-parasitic cysts of the liver is mostly palliative such as aspiration of the content, suture and closure, internal or external drainage, marsupialization, and unroofing of the cyst, while total excision of the entire cyst, which sometimes necessitates hepatic resection, is not usually recommended.3, 6, 9 The results of these lesser procedures have been acceptable, favoring those conservative procedures. This paper presents three cases with carcinoma arising in the cysts of the liver. Review of the present cases with five comparable cases appearing in the literature revealed that young female population and left lobe of the liver are frequently involved. Hence the general trend for palliative procedures should be reassessed. Possible malignancy should be considered when a patient belongs to this category, the cystic content is not clear, or the cystic wall presents irregular texture with nodules. The carcinomatous changes would have been of higher incidence than reported if the entire cysts had been more carefully examined.

Survival for 145 days with a total artificial heart.

A calf into which a biolized, total artificial heart (TAH) had been implanted survived for 145 days. All measured physiological parameters except central venous pressure (CVP) were back to normal one month after implantation, and thereafter the animal's physiological development was similar to that of a normal calf. The intimal weight, which was 96 kilograms at implantation, reached 190 kilogram at the end of experiment, with a daily gain rate of 0.9 kilogram per day. After the nineteenth postoperative week, signs of congestive heart failure appeared, such as high venous pressure, ascites, and enlarged liver although the calf outwardly appeared well. On postoperative day 146, the animal started foaming at the mouth, and a convulsion occurred; then, the experiment was terminated after 3,494 hours of pumping. At autopsy, there were acute bilateral bronchopneumonia involving mostly both upper lobes, pulmonary edema, slight chronic pneumonitis, and hepatomegaly. There were no serious thrombotic deposits inside the cardiac prosthesis.

145 days survival of calf with total artificial heart (TAH).

A calf with a biolized TAH was maintained in reasonably good physiological condition for a major portion of a 145-day experiment as indicated by near normal growth rate and laboratory data. The mechanism for the high CVP and CBV at the beginning and end of the experiment were correlated with protein metabolism. No evidence of thromboembolism was found at autopsy and only modest congestion with minimal parenchymal damage was observed. The cardiac prosthesis itself was free of thrombus, although slight calcification was found on the valves and diaphragms. The pump performance, however, remained satisfactory. In addition, endothelialization appeared to be starting on a portion on the inside of the pump.

Application of aldehyde treatments to cardiovascular devices.

Biolized natural and synthetic materials represent a new class of materials. Aldehyde treatment of natural tissue creates cross-links in the collagen molecules while retaining mechanical strength and collagen structure. Synthetic polymers can also be biolized by the addition of protein and aldehyde treatment. Cross-linked materials such as these are resistant to degradation by proteolytic enzymes. The procedure for the aldehyde treatment of natural tissue and protein polymer composites has been established, and in vivo and in vitro studies have demonstrated an improved blood compatibility. Long-term survival with TAH and LVAD implanted animals has shown the successful application of these materials without the use of anticoagulants. Pseudoneointima growth occurs on these surfaces, and results to date indicate growth stabilization within 2 wks of impalntation. Studies are currently underway to fully characterize the pseudoneointima formed on the biolized surfaces of cardiac prostheses.