The Nb rat: prostatic adenocarcinoma model. The Noble rat prostatic adenocarcinoma is an animal model which is used to study human prostatic carcinoma. It mimics the human condition in histology, in biochemistry with the production of acid phosphatase, and in tumor growth which is relatively constant. Additionally, it is hormonally dependent and metastasizes. This animal model should provide an avenue for the evaluation of cytotoxic chemotherapeutic agents heretofore receiving little attention from the urologic community.


The response of the renal pelvis to infection: a scanning electron microscopic study. The response of the rat renal pelvis to Pseudomonas aeruginosa infection was examined using scanning electron microscopy. Bacterial attachment was noted in regions of microplical alteration with subsequent strand formation and epithelial cell exfoliation. These observations support the hypothesis that bacterial invasion initiates defense mechanisms in the renal pelvis similar to those noted in bladders, namely, membrane alteration with increased bacterial attachment, strand formation with further bacterial entrapment, and exfoliation of bacteria-laden epithelial cells which are eliminated via voiding.


The ultrastructural effect of distension on the neuromuscular apparatus of the urinary bladder. The recent clinical success of distension therapy in the treatment of idiopathic detrusor hyperreflexia has prompted this ultrastructural study. The neuromuscular pathways of conduction in the rat and rabbit urinary bladder wall were examined using perfusion fixation techniques. The effects of short-term and prolonged distension were studied at frequent intervals for 4 months in treated and control animals. Prolonged distension did not alter either smooth muscle architecture or intercellular contact points. A transient phase of nerve degeneration in bladder muscle consisting of axonal swelling and lysis of organelles was observed. A quantitative estimate of nerve injury and regeneration was compiled using pooled histograms. Possible physiologic mechanisms of injury and clinical conclusions are discussed.


Retinyl acetate prophylaxis in cancer of the urinary bladder. Virgin female C3H/He mice (275) were fed a ration that included 0.1 per cent N-4-(5-nitro-2-furyl)-2-thiazolyl formamide (FANFT) and retinyl acetate. The mice were divided into five groups with Groups 1 through 4 receiving 300, 600, 1200, and 2400 IU of retinyl acetate per kilogram of feed plus FANFT respectively. Group 5 received 300 IU per kg of feed and no FANFT as a control. After 45 weeks the bladders were removed and inspected for neoplasms. There were no tumors found in any of the mice from Group 5. It was found that retinyl acetate significantly inhibited the formation of squamous cell tumors at all levels and inhibited transitional cell carcinoma when given as 600 IU per kilogram of feed.


Adenosine deaminase activity in a transplantable murine renal adenocarcinoma. Adenosine deaminase (ADA) activity was studied in tumor tissue, spleen, and bone marrow of animals bearing a transplantable murine renal carcinoma. Tumor tissue and splenic ADA specific activities were elevated in these animals when compared to controls although splenic specific activity subsequently declined with increasing tumor burden. Marrow ADA specific activity was the same in tumor bearing animals as in controls regardless of variations in tumor size. Studies of this enzyme may offer insight into salvage pathways of purine biosynthesis thereby suggesting exploitable targets for chemotherapy.


Pharmacokinetic studies of oxalate in man. The observation of the plasma concentrations after rapid intravenous administration of inulin and 14C-oxalate to six normal and one hyperoxaluric subjects allowed estimations of the half-life of elimination of oxalate (mean 91.7 min), volume of distribution of oxalate (mean = 32.5 liters), calculation of the miscible oxalate pool (mean 3.7 mg), of the plasma oxalate concentration (mean = 11.1 microgram per 100 ml), of the oxalate clearance (252 ml per min), and the determination of the oxalate turnover rate (0.027 mg per min). The 14C-oxalate/inulin clearance ratio was 2.44 implying an additional excretion mechanism apart from glomerular filtration. Inasmuch as 97.2 per cent of the administered tracer were recovered unchanged in urine, nonrenal loss of oxalate is not an appreciable factor. The data of one hyperoxaluric patient are compared with those of the healthy subjects.


Laser investigations of the strictured dog urethra. The effects of the Neodymium:YAG laser beam on normal as well as on granulation tissue of the dog urethra were investigated in vitro. To determine the effectiveness of the laser beam special attention was paid to the size of the local spot and the pulse duration. After determination of the most favorable diameter of the focus and the pulse duration of the laser beam, endoscopic laser examinations were performed on normal and experimentally strictured urethrae in dogs. A suitable laser endoscope was designed and a surgical technique for laser-beam treatment of the strictured urethra was developed. The postoperative follow-up period was up to 6 weeks. There was no need for postoperative urinary diversion. Neither bleeding nor perforations were observed.


Influenza virus: appearance of high mouse-neurovirulent recombinants. Recombinants from two influenza A strains that lacked mouse neurovirulence were tested, along with their parent strains, for mouse neurovirulence and for the ability to propagate in dissociated mouse embryo brain cells. The parents used were (i) strain A/Rostock/34 (FPV) (Hav1N1), with a high chicken neurovirulence, and (ii) the mouse-lung-adapted human strain Engl/1/61 (H2N2), lacking neurovirulence. In some of the recombinants high mouse neurovirulence could be detected after intracerebral inoculation of low virus doses. There was neither a correlation between surface antigen and neurovirulence nor between neurovirulence and mouse lung virulence in our system, although neurovirulence was only found in strains with Hav1 hemagglutinin. There was an association between replication in mouse embryo brain cells in culture and high mouse neurovirulence.


Efficient ribosome binding of brome mosaic virus (BMV) RNA4 contributes to its ability to outcompete the other BMV RNAs for translation. Analysis of translation products synthesized in vitro in the presence of a mixture of brome mosaic virus (BMV) RNAs 1, 2, 3, and 4 usually shows a predominance of coat protein, coded by RNA4. The proportion of products directed by RNAs 1 and 2 decreased at saturating concentrations of BMV RNA and in the presence of 7-methylguanosine 5'-phosphate (m7G5'p) at nonsaturating concentrations of RNA. No differences in the relative proportions of the various proteins produced were detected in the presence of T-2 toxin, which interferes with the formation of the first peptidyl bond rather than competing for initiation factor, as has been suggested for m7G5'p. These data show that the ability of BMV RNA4 to outcompete BMV RNAs 1, 2, and 3 results, at least in part, from its ability to bind more efficiently to ribosomes.


Plaque assay of baculoviruses employing an agarose-nutrient overlay. Four baculoviruses produced visible plaques in the Spodoptera frugiperda cell line when cell monolayers were infected and overlayed with a simplified, agarose-nutrient formulation. Macroscopic plaques were first detected 4 days postinoculation, and by 10 days plaques ranging from 0.5 to 3 mm in diameter were seen. Dose-response experiments indicated that a single particle initiated the formation of a plaque, because a linear response was demonstrated with increased dosage.


Classification of human adenoviruses by SDS-polyacrylamide gel electrophoresis of structural polypeptides. The virion polypeptide compositions of 15 human adenovirus types, representing all members of subgroups B and C and selected members of subgroups A and D, have been analyzed by SDS-PAGE. The capsid polypeptides II, III, IIIa and IV displayed a pattern of apparent molecular weights which was characteristic for each adenovirus serotype analyzed. Adenovirus serotypes belonging to the same subgroup were also demonstrated to share an apparent molecular weight pattern of the internal structural polypeptides in the following manner: Subgroup A:V-resolved into two bands of 46--48.5 K and 51--51.5 K; VI-25.5 K; and VII-18 K. Subgroup B:V-53.5--54.5 K; VI-24 K; and VII-18 K. Subgroup C:V-48.5 K; VI-24 K; and VII-18.5 K. Subgroup D:V-50.5 K; VI-23.5 K; and VII-18 K. Adenovirus type 4: V-48 K; VI-24.5 K; and VII-18 K. The structural polypeptides represent a major portion of the adenovirus gene products. It is therefore proposed that the apparent molecular weight differences of viral polypeptides be used (i) to confirm the identification of an adenovirus type, and (ii) for subgroup classification of adenoviruses and, consequently, that adenovirus type 4 should be classified as a member of a separate subgroup designated E.


Water-bone transmission of influenza A viruses? The isolation of influenza A viruses from unconcentrated lake water and from fecal samples on the shore of these lakes is reported for the first time. Influenza A viruses, representative of most of the major antigenic subtypes, co-circulate in ducks on the lakes.


Radioimmunoassay for albumin-binding sites associated with HBsAg: correlation of results with the presence of e-antigen in serum. A radioimmunoassay for albumin-binding sites associated with hepatitis B surface antigen (HBsAg) is described. Polystyrene beads coated with glutaraldehyde-polymerized human serum albumin were incubated with diluted specimens of HBsAg-positive sera which contained either e-antigen (HBeAg) or anti-HBe or were negative for both HBeAg and anti-HBe as determined by rheophoresis. The quantity of HBsAg attached to the beads, determined by measuring the amount of 125I-labeled anti-HBs adsorbed during a subsequent incubation step, was related to the presence of HBeAg in the specimens.


Evidence for a single infectious species of potato spindle tuber viroid. Potato spindle tuber viroid (PSTV) has been separated into two electrophoretic species on polyacrylamide gels containing 8 M urea at 60 degrees. Only the slower-migrating electrophoretic form was infectious and is presumably the circular form of the molecule. Both forms had similar base compositions with a 55-58% GC content, similar to the citrus exocortis viroid. Labeling studies in vivo suggest that the viroid is not methylated.


Suggestions for the classification and nomenclature of helical plant viruses. Helical plant viruses are arranged into four families: Flexiviridae, Tobamoviridae, Hordeiviridae, and Tobraviridae. Further division of these families into genera and subgenera is shown. A family-specific, constant amount of RNA per unit virion length and a distinct radial location of RNA from the center of the helix are considered as basic criteria for establishment of these families.


Formation of cross-linked molecules during vaccinia DNA replication. The appearance of cross-linked DNA molecules during the replication of vaccinia virus DNA was investigated. Conversion of full size DNA molecules to cross-linked molecules occurs several hours after completion of DNA synthesis. Inhibition of RNA synthesis by actinomycin D did not prevent this transition; however, interruption of protein synthesis by streptovitacin A resulted in partial inhibition of the process.


Purification and partial characterization of maize mosaic virus. Leaf extracts containing maize mosaic virus (MMV) were purified by treatment with activated charcoal followed by filtration through Celite. Virus was concentrated from the filtrate by sedimentation onto a sucrose pad and then further purified by centrifugation in a 20--50% continuous sucrose density gradient. Buoyant density of virus particles in sucrose solution was 1.181 g/ml. Nucleic acid extracted from virus preparations reacted with orcinol and behaved like RNA in Pederson's diphenylamine test. It was digested by pancreatic RNase and apparently is single-stranded RNA. Virus preparations also contained lipids. The morphology and constituents of MMV particles are therefore those expected for a rhabdovirus.


Physicochemical characterization of a small RNA virus associated with baculovirus infection in Trichoplusia ni. An RNA virus was isolated from Trichoplusia ni larvae infected with inoculum of the baculovirus of Autographa californica. Physicochemical characterization has revealed a 35-nm virus with a density in CsCl of 1.3 g/cm3 and a sedimentation coefficient of 200S. The virus consisted of 10--15% single-stranded RNA with an apparent molecular weight of 1.9 x 10(6) and coat protein with a subunit size of 67,000. The virus is similar to the Nudaurelia capensis beta virus and also possesses some interesting similarities to the mammalian caliciviruses. The proposed cryptogram of the virus is R/1:1.9/12:S/S:I/O.


[60 cases of stenosis of the thoracic esophagus. Surgical technic]. Thoracic esophageal stenosis most frequently is seen in reflux disease as sequelae of endobrachyesophagus. In 10% of these cases later on an adenocarcinoma will be proven. Of special therapeutic interest in this field is the relationship with the Zollinger-Ellison syndrome, with scleroderma and with juvenile acid burns of the esophagus. As operative therapy in the benign state of reflux disease we advise conservative, organ-preserving, functional surgery with peroperative dilatation, abdominal fundoplication and, if necessary, vagotomy. Long-term follow-up results are demonstrated.


[Perforations on peptic ulcers of the esophagus]. The term "atraumatic perforation of the oesophagus" which is suggested by ABBOTT covers several entities. HEBERER and RIOULT draw a distinction between spontaneous ruptures, which have their seat in a macroscopically healthy zone of the oesophagus, and perforations, which are the final consequence of a constant, progressive process of parietal erosion. Such a perforation may be due to complications of the peptic disease of the oesophagus. The transition from the perforating or intramural ulcer to the disintegration of the entire wall, with the formation of a mediastinal abscess, is progressive. The seriousness of the affection results from the perforation of this abscess into a neighbouring organ or cavity (mediastinum, lung, pericardium, pleura). In most cases, however, the perforation is closed by reactional mediastinitis, which also limits the seriousness of the phenomenon and permits a comparatively mild form of treatment (drainage).--Out of six cases of atraumatic perforation, we have observed two perforations on peptic oesophagitis: one of them exceptional, perforated into the aorta with immediate death, and the other perforated into the mediastinum and cured after resection of the oesophagus.


[Esophageal leiomyomas. Apropos of 16 cases]. 16 cases of oesophageal leiomyomas are reported; diagnostic difficulties and important surgical consequences are discussed. The surgical procedures must be as soon as possible, and should be enucleation of the lesion. The results are excellent. In three cases postoperative oesophageal spasms and retrosternal pains needed secondary medical treatment.


[Necrosis of the lesser curve of the stomach after proximal selective vagotomy]. Necrosis of lesser curve of stomach after proximal selective vagotomy is a specific but rare hazard of this operation. Anatomical and pathophysiological findings speak in favour of ischaemic pathogenesis, even if a peptic factor may exceptionally play a role in some cases. Local tissue trauma enhances the risk of later sloughing of lesser curve. Three own reported cases without fatality point out the importance of previous massive ulcer bleeding with hypovolemic shock and severe anaemia. Early recognition and immediate reoperation only may avoid an otherwise fatal course. Simple oversewing has proved successful in treating even large lesions. Careful, atraumatic technique of skeletonization and reperitonealization of lesser curve help to prevent this life threatening complication.


[Proximal selective vagotomy with or without pyloroplasty?]. Proximal gastric vagotomy without drainage is the operation of choice for uncomplicated duodenal ulcer. There are few contraindications for PGV as uremia, diabetes, hypertension, age over 65 years and a history of splenectomy. Only in cases of severe pyloric stenosis or bleeding ulcer or perforation in the pyloric area, a pyloroplasty should be added. The Wangensteen pyloroplasty is a safe drainage procedure and especially recommended in case of extensive scarring of the pylorus.


[Value of abdominal ultrasonography for indication of cholecystectomy]. A review of the gall-bladder content of the 193 patients that underwent a cholecystectomy at the University Hospital of Lausanne, Switzerland, in 1977, was done in order to find out the diagnostic accuracy of sonographic examination of the gall-bladder, oral cholecystography and intravenous cholangiography. Sonographic examination of the gall-bladder proofed to be accurate in 92.5% of 109 cases. We found a false negative rate of 4.5% (5 cases) and a false positive rate of 1% (1 case) as well as an indeterminate rate of 2% (2 cases). In 37% (46 cases) of the 124 patients that underwent oral cholecystography and/or intravenous cholangiography the gall-bladder was not visualized. Sonographic examination of gall-bladder is free of side effects and can be done on emergency, on jaundiced or pregnant patients, and it is enough reliable for the surgeon to make a decision to operate the gall-bladder. Low cost of the procedure is another point in favour of sonographic examination of the gall-bladder.


[Papillotomy or papilloplasty? Clinical and endoscopic follow-up after papillary surgery]. We controlled patients after surgical incision of the papilla for papillary stone or papillary stenosis. 131 were examined clinically, 39 of them additionally with ERCP. The endoscopy showed that simple cutting of the papilla (i.e. papillotomy) often resulted in a restenosis because of accretion of the incised duodenal mucosa. This was prevented by the additional suture of the choledochal and duodenal mucosa (i.e. papilloplasty). The clinical results, however, were equally good with both surgical techniques. This rises the question in view of the striking endoscopical results, whether the clinical examination alone is an adequate criterion to detect stasis disorders of the liver and the pancreas. The results confirm us to perform, when surgical incision of the papilla is necessary, rather a papilloplasty than a papillotomy.


[Indication of delayed surgery in the postacute phase of hemorrhagic necrotizing pancreatitis]. In acute hemorrhagic-necrotizing pancreatitis partial necrosis with good response to conservative therapy can be differentiated from extensive necrosis with no response to conservative therapy. In case of surviving the acute phase sequester and abscess are often to be seen in the "postacute phase", after 10-14 days. The indications for the "delayed operation" in this postacute phase are development of a palpable mass together with clinical deterioration and other complications. The surgical procedure consists of digital removal of necrotic tissue (sequestrotomy), abscess incision and resection. Since 1971 84 of 93 patients with a postacute pancreatitis were operated, two third survived.


[Surgical treatment of hyperlipidemia by partial ileal by-pass]. This study is composed of 7 hyperlipidaemic patients (type II, n = 4; type IV, n = 3), aged 33 to 59 years. These patients underwent a partial ileal by-pass, and the immediate post-operative period was uneventful. Six months post-operatively, serum levels of cholesterol and triglycerides were 45 and 44% of the pre-operative values. Serum electrolytes remained unchanged as well as liver function tests. In the first few days post-operatively, diarrhoea was a frequent complaint; by the 6th post-operative month, the stools numbered 2-3 per day in patients not receiving any medical treatment. Average weight loss was 6 +/- 1.3%. These preliminary results may confirm that partial ileal by-pass produces favourable metabolic changes. However, screening for late complications is to be further investigated, and the effectiveness of the operative procedure must still be evaluated.


[Rectosigmoidovesical fistula].

[Rectosigmoidovesical fistula]. The symptomatology of vesico-colonic fistulae is presented and their differential diagnosis discussed. Management should be tailored to the individual case. Whenever possible, we perform a radical primary operation. An additional by-pass colostomy is indicated if it is felt that the integrity of the anastomosis is jeopardised.


[Colonic perforation with special reference to spontaneous ileo-colic perforation]. Aetiology of colonic perforation is reviewed and discussed. 8 cases of "spontaneous" ileo-colic perforations observed between 1975 and 1977 are presented. Two of these patients had recently undergone appendectomy, and 4 others showed a simultaneous distal carcinoma of the large bowel. Histo-pathological evaluation did not reveal the cause of intestinal perforation. Operative treatment and results are given.


[Sigmoid colon injuries caused by blunt abdominal trauma]. Circumferential seromuscular stripping of the sigmoid colon with denudation of the mucosal tube over a length of 4 to 10 cm has been observed in four patients after blunt abdominal trauma. Three of the patients had been driving a motor car, with two of them wearing safety belts. One patient was injured when he was caught between steel bars at work. In two cases the sigmoid lesion was associated with a tear in the mesoileum, leading to ischemic bowel necrosis in one. In one patient the lesion was found 5 days after the accident when he presented the symptoms of diffuse peritonitis caused by secondary perforation of the denuded mucosal tube. Treatment consisted in resection, anastomosis and proximal colostomy in two, and resection with end colostomy and distal mucous fistula in the remaining two cases.--As long as perforation of the denuded mucosa has not occurred, symptoms may be minimal. Liberal use of peritoneal lavage in polytraumatized patients is therefore important in order to minimize the likelihood of overlooking this type of bowel lesion.


[Treatment of anal fissure by lateral subcutaneous internal sphincterotomy]. 50 patients have been followed up after lateral internal sphincterotomy for chronic anal fissure. The results are excellent. This operation has some advantages over against the posterior sphincterotomy or the sphincter stretching and should replace these latter procedures in the treatment of chronic anal fissure. The lateral internal sphincterotomy may probably be applied for other benign anal lesions.


[Starch peritonitis. Apropos of 13 cases]. First known as an inert and harmless substance, starch powder has been used from 1950 by surgical gloves factories in place of talcum. Its innocuousness is now being denied, but many surgeons still do not know its adverse side effects. 13 cases of glove starch peritonitis, seen between 1971 and 1978, allow the authors to review clinical signs, diagnostic and treatment possibilities as well as the risk of a new operation. To limit the quantity of starch powder introduced into the peritoneal cavity is so far the only mean to prevent this complication and to compel the surgical team to strict discipline.


[Prevention of parietal septic complications by irrigation of the surgical wound]. In a randomized group of 162 patients, undergoing appendicectomy, irrigation of the wound with saline alone versus saline and antibiotics, shows a statistically significant smaller incidence of wound infection than in a comparative non irrigated group.


[Burn mortality and morbidity: experience of the burn center in Zürich from 1967 to 1977]. Ten years of burn treatment in a burn center are compared to the ten years prior to 1967: --Survival has increased markedly, due to aggressive local treatment with débridement, topical ointments, biologic dressings, and rapid closure of wounds. --Functional and cosmetic results have improved since tangential excision and immediate autografting are routinely performed. --Favourable results with shorter duration of hospitalization are obtained if doctors and nurses can acquire and apply their knowledge in many cases as is possible in a burn center.


[Superselective parathyroid hormone determination--a reliable diagnosis of the location of endocrinological active parathyroid neoplasms]. The importance of superselective catheterization of the small jugular veins for PTH-radioimmunoassay is shown as an improvement of the preoperative localization technique. Since May 1976 in 20 patients the diagnosis of pHPT could be established. Our own experiences with the method of small vein sampling combined with the radioimmunoassay demonstrate that a reliable localization of overactive parathyroid tumours is possible.


[Various amounts of estrogen receptors in breast neoplasms]. An agar gel electrophoresis was performed to demonstrate various amounts of estrogen receptors of three different topics in 38 mammary carcinomas. A good correlation between estrogen receptors in primary mammary carcinomas and axillary lymph node metastases could be found.


[Does every hospital need a catastrophe plan?]. The organisation of any hospital to manage the medical supply in case of general disaster should guarantee the maximum effectiveness. The increase of output has to be realised on the emergency ward, OR, JCU and on the general wards. The planning should include the management, the competence and the coordination inside and outside the hospital.


[OPS--a new surgical planning system].

[OPS--a new surgical planning system]. Due to increasing specialization the organization of units of operating theatres in big and middle-sized hospitals becomes more and more difficult. The capacity of operating rooms is the limiting factor in most hospitals, a careful and coordinated planning is therefore of great importance. We describe a new kind of planning system, which has been developed in our department. Experience has been excellent so far.


[Repair of recurrent very large inguinal hernias with intraabdominal prosthesis]. The use of a prosthesis in the repair of very large inguinal hernias as well as in multiple recurrences is often advocated. A new material is presented: the Rhodergon is a sheet of polyester which is placed through an intraabdominal extraperitoneal approach in order to cover the defect of the wall. One of its surfaces is velvet-like and induces adhesions with the edge of the abdominal defect that will be responsible for the strength of the repair. The other surface is covered with silicone and prevents adhesions with the peritoneum. Our first results with this type of repair are described and so far seem extremely satisfactory.


[Acute arterial obstructions of the limbs in the aged]. 131 cases of acute arterial obstruction were treated during 1973-1977, 70 (52%) of these cases were seen in patients of over 70 years of age and 20 out of them died during the first two months of the surgical treatment, 8 patients were amputed. The long term evaluation of 40 patients who survived for more than two months after the surgical treatment shows a very high mortality (14 of the 40 surviving patients died during the first two years of the surgical treatment). Satisfactory results were obtained in 18 patients. The long term results after arterial obstruction in old patients has high mortality and the number of amputations is directly proportional to the delay before commencement of the surgical treatment.


[Therapeutic difficulties encountered in the treatment of fractures of the acetabulum. Retrospective study of 44 cases of osteosynthesis of the acetabulum]. A retrospective study shows several factors that influence the ultimate prognosis of fracture-dislocations of the hip. The magnitude of the initial injury worsens the prognosis as far as functional recovery and residual pain are concerned. The most difficult cases are those with anterior and posterior fractures of the acetabulum. The quality of reduction is often imperfect and the techniques of surgical repair are very difficult and time consuming. Provided there is no associated lesion of the sciatic nerve, the more simple fracture (transverse and posterior) gives better operation results. Reduction is often entirely successful if the surgical technique is adequate. Despite proper treatment, some patients develop aseptic necrosis of the femoral head and are left with most severe sequela.


[Clinical experience in 80 patients with intramedullary trochanteric nailing]. Report on the clinical experience in 80 patients with the intramedullary trochanteric nailfixation according to KUNTSCHER of pertrochanteric, or subtrochanteric femur fractures. This procedure in our opinion enlarges the operative methods for treatment of fractures of the trochanteric region in elderly patients with a high operative risk.


[The combination of fractures of the lower extremity and lesion of the posterior cruciate ligament]. In 400 fractures of the upper and lower leg, 7 patients had an additional lesion of the posterior cruciate ligament. This combination occurs in people, who had a high energy frontal impact in the sitting position. The ligamenteous injury is often overlooked because of multiple injuries and difficulty in examination of a knee joint on a fractured leg. In high velocity injuries of the lower extremity, knee and hip joint injuries must not be overlooked. The diagnosis of knee injury can be made by inspection and palpation of a joint effusion and by a knee joint X-ray, which shows an avulsion fracture of the dorsal tibial spine. Repair is recommended within the first 3 weeks if possible. Later, scarring in the subluxed position and osteoporosis of the fragment will make a reconstruction very difficult.


Bile duct carcinoma associated with an anomaly in the bile duct. This paper reports a case of bile duct carcinoma associated with an extra-hepatic bile duct anomaly. In the present case, the anomaly induced an uncorrect preoperative diagnosis and obliged to change the surgical procedures during operation. A correct preoperative diagnosis was difficult to make in the present case, when the anomaly in the biliary system was not kept in mind.


[Relaparotomy or endoscopic surgery in complications after bile duct surgery]. The number of cases in bile duct surgery has continuously increased since second world war. Correcting operation is necessary in 5 to 7% after cholecystectomy. Since 1973 more and more the endoscopic treatment takes place of surgery. Typical complications can be divided into two groups belonging to their manifestation. Early complications are due to an insufficient intraoperative diagnosis or injuries of bile tract. Late complications comprehend successive development of duct stenosis and cholangitis. The therapeutic possibilities of endoscopic papillotomy are directed only to the papilla and near-by sections of the choledochus, while surgical treatment is due to the more difficult performance of iatrogenic lesions. The indications to each method are shown and discussed by 138 surgical and 714 endoscopic treatments.


[Crohn's disease of the appendix].

[Crohn's disease of the appendix]. 22 cases of primary involvement of the appendix in Crohn's disease are reported in the literature. During the last 3 years we observed 4 cases. In three patients typical clinical signs of acute appendicitis were present. One case was found occasionally. The follow-up was 5-30 months. During this time none of our patients showed symptoms of granulomatous enterocolitis in other regions of the bowel. Reviewing the literature 3 of 26 patients (11.5%) showed lesions typical for Crohn's disease elsewhere in the intestine 3-48 months after appendectomy. But two cases had a follow-up of 5 years which time is considered indispensable before one can admit that the disease is really limited to the appendix.


[Turning-point of the bowel preparation?]. The whole-gut irrigation more and more carries through to be a favourite method for preoperative large bowel preparation. In 1975 it was used only by 3 hospitals in Switzerland, which resulted from our investigations about large bowel preparation. This new procedure brings advantages such as complete cleaning of the gastrointestinal tract, shortening of the preoperative hospitalisation time, retarded beginning of postoperative bowel function, no catabolic situation of patients' metabolism at operation-day, good compatibility and simple handling, according to our experience with 75 cases. There are not any disorders of the fluid and electrolyte balance except a small intake of water without clinical importance. The indication for whole-gut irrigation is given in all patients, who are subjected to an operation on the colon and rectum except the emergencies. Ileus, toxic megacolon and perforation of the large bowel are considered absolute contraindications. Severe large bowel stenosis, cardiac and renal insufficiency are relative contraindications only.


Maturation of renal function in full-term and premature neonates. The development of renal function was studied in neonates with gestational ages ranging from 28 to 43 weeks. The effect of gestational age on the maturation of renal function was assessed in newborn infants studied during the first 72 h of life. Postnatal maturation was examined during the first 3 weeks of life. None of these neonates presented any cardiopulmonary or renal disturbances. Inulin and PAH were used as markers of glomerular filtration rate and effective renal plasma flow, respectively. From the 28th to the 35th week of gestation, there is a progressive increase in inulin and PAH clearances (r = 0.93; p less than 0.001 and r = 0.81; p less than 0.005, respectively), which then level off up to the end of gestation. A linear increase in systemic blood pressure is observed during the last 12 weeks of gestation (r = 0.88; p less than 0.001). There is a positive significant correlation between the rise in blood pressure and inulin or PAH clearance (p less than 0.005). Postnatal maturation of renal function is similar in preterm and term neonates. In both groups birth is followed by a sharp rise in inulin and PAH clearances, a twofold increase of these parameters being observed during the first two weeks of life.


[Effect of previous breast feeding and early diagnosis on the course of cow's milk protein intolerance]. The clinical course of 21 children with cow's milk protein intolerance (CMPI) who were treated at the Children's Hospital of Zurich during the years of 1963-1977 was analysed in retrospect. Oral cow's milk challenges undertaken 3 weeks after the beginning of the treatment or later did not lengthen the apparent duration of the disease. Children who had initially been breast fed during at least 10 days, and those who were treated within 2 weeks after the appearance of the first symptoms with a cow's milk-free diet presented a milder and shorter course of CMPI. Severely undernourished or dehydrated patients did not differ from others in the course or duration of the disease, and patients with bloody stools showed a less serious but not shorter course of the CMPI.


Anemic necrosis of the liver after umbilical vein catheterization. We report 22 cases of hepatic necrosis following catheterization of the umbilical vein. The morphology of the necrotic lesions, which were confined to the left lobe in 21 patients, was that of an anemic infarct. Their causes are multiple and include intimal damage during the insertion of the catheter with consecutive thrombosis of portal vein branches, the postpartum immaturity of the arterial hepatic oxygen supply, postpartum hypoxia due to respiratory failure of cerebral or pulmonary origin, obstruction of portal venous flow through the catheter as such, the postpartum involution of the left hepatic lobe, and prolonged periods of catheterization. Toxic damage of the hepatic parenchyma due to an infusion of Tris buffer through the catheter into the portal venous system is a potential additional factor.


Hypodipsia-hypernatremia syndrome.

Hypodipsia-hypernatremia syndrome. The pathogenesis of the rare hypernatremia, usually described in the literature as "neurogenic" or "essential" hypernatremia, consists of defective thirst mechanism either alone or in combination with impaired osmoregulation of ADH release. As etiology, disturbances of the neoplastic, vascular and degenerative type and malformations in the hypothalamic area are known. In patients with the hypodipsia-hypernatremia syndrome, dysfunction of the anterior pituitary lobe, obesity, abnormal regulation of body temperature, psychomotor retardation and episodic muscular weakness are frequently encountered as additional abnormalities. A 6-year-old patient is described with hypodipsia-hypernatremia syndrome manifest for 3 years. Besides hypernatremia, hypodipsia and the relative insensitivity of the osmoreceptors regulating ADH release, elevated body temperature, polyphagia and obesity, partial hypothalamic-hypophyseal dysfunction, lethargy and psychomotor retardation are the principal findings. An inflammatory lesion or one occupying an intracranial space was not demonstrable until now. Under forced water intake and hypocaloric diet the patient has progressed well with nearly complete normalization of the hypernatremia, body temperature and obesity.


[Pulmonary valvular subatresia with intact interventricular septum in a newborn infant: role of prostaglandins in the immediate postoperative period]. The case is reported of a newborn with pulmonary valvular subatresia and intact interventricular septum and moderate right ventricular hypoplasia, treated with pulmonary commissurotomy alone. In the immediate postoperative period the ductus arteriosus closed, resulting in deep hypoxia and acidosis. With the infusion of PG E2 the ductus reopened and the baby's condition improved markedly. The ductus was kept open for 20 days, during which time adaptation of the right ventricle must have occurred. Indeed, after definitive spontaneous closure of the ductus, right ventricular output proved sufficient to insure satisfactory pulmonary perfusion.


[Renal lesions in congenital syphilis. A case report]. The authors describe a case of congenital syphilitic nephropathy in an infant. The review of the reports in the literature shows many similarities of this congenital disease with the syphilitic nephrotic syndrome in the adult. The prognosis is favorable provided the diagnosis is made sufficiently early. Histological studies utilizing light-, electron- and immunofluorescence-microscopy gave results consistent with extramembranous glomerulonephritis with possible evolution towards endo- and extracapillary proliferative glomerulonephritis. This is one of the rare renal diseases with a known antigen.


Comparative studies on the nature of purified cytomembranes of the rabbit parotid gland. Centrifugation procedures have been evolved for isolating purified samples of rough endoplasmic reticulum, Golgi, zymogen granule and plasmalemmal membranes from homogenates of rabbit parotid gland tissue. The purification process was monitored using morphometry and enzyme and chemical marker assays. The membrane preparations were analysed by sodium dodecylsuphate (SDS) polyacrylamide gel electrophoresis, quantitative phospholipid thin layer chromatography and by enrichment studies. The results were used to evaluate various possible general models for the behaviour of membranes during the secretory cycle of parotid acinar cells.


The ultrastructural localization of the enzymes related to steroid hormone metabolism in the guinea-pig testis. A study of the ultrastructural localization of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD), glucose-6-phosphate dehydrogenase (G-6-PD), beta-hydroxybutyrate dehydrogenase (beta-HBD), NADH diaphorase (NADH-D) and NADPH diaphorase (NADPH-D) in the guinea-pig testis is reported. The procedures employed included short immersion or perfusion fixation with aldehydes followed by incubation of small blocks in a tetrazolium salt or a ferricyanide medium. The effects of incubation conditions were investigated, and a reaction medium for the ultracytochemical demonstration of 11 beta-HSD is described. Using suitable controls, evidence for the specificity of the cytochemical reactions is presented. It was found that all the enzymes studied were present in both the Leydig and Sertoli cells of the guinea-pig testis and that the intracellular distribution pattern for each enzyme was independent of the cell type. Using tetrazolium salt techniques, both 3 beta-HSD and 11 beta-HSD activities were localized on or in membranes of smooth endoplasmic reticulum and within the mitochondria. With the ferricyanide techniques, G-6-PD activity was found to be associated mainly with the smooth endoplasmic reticulum membranes, while beta-HBD activity was limited to mitochondria. With both the tetrazolium salt and ferricyanide techniques, the reaction products for NADH-D and NADPH-D activities showed localizations which were similar to those observed for the steroid dehydrogenases.


Histochemical characterization of monoamine oxidase in ependyma of rat hypothalamus. Monoamine oxidase (MAO) activity has been demonstrated histochemically in rat hypothalamic ependyma using the sulphate-tetrazolium and coupled peroxidatic techniques with tryptamine, tyramine, 5-hydroxytryptamine and benzylamine as substrates. Both methods were applied to cryostat sections with and without exposure to selective amine oxidase inhibitors, including the selective A-MAO inhibitor clorgyline, and the B-MAO inhibitor deprenyl. Our results show that both cuboidal-columnar and tanycyte ependyma contain one or more forms of MAO not generally present in the hypothalamus. It is suggested that ependymal MAO may form an amine-barrier system modulating the movement and effect within the hypothalamus of specific cerebrospinal fluid or blood monoamines.


Immunohistochemical localization of erythropoietin in the rat and mouse submandibular gland. There is a great deal of evidence to indicate that organs other than the kidney are involved in erythropoietin production. In this paper, it is reported that erythropoietin has been localized with an immunocytochemical method in the granular ducts of the submandibular glands of the rat and mouse.


Multi-element analysis of the rat hippocampus by proton induced X-ray emission spectroscopy (phosphorus, sulphur, chlorine, potassium, calcium, iron, zinc, copper, lead, bromine, and rubidium). A technique for multi-element analysis of brain tissue by proton induced X-ray emission spectroscopy (PIXE) is described and data from analysis of fixed and unfixed samples from rat hippocampus, neocortex, amygdala, and spinal cord is presented and commented on. The atoms present in the tissue are bombarded with protons which causes the ejection of electrons from the inner shells. When the "holes" are refilled with electrons from outer shells, X-ray quanta characteristic for each element are emitted. Using a high resolution energy dispersive detector a complete X-ray spectrum of the specimen can be recorded in a single measurement. Detection limits less than or approximately 5 p.p.m. of dry matter are obtained for most elements with atomic number greater than 14 (silicon). Around 13 elements were found in concentrations above the detection limits. The grand means for non-fixed hippocampi were e.g. for Zn - 120 p.p.m.; Rb - 20 p.p.m.; Fe - 150 p.p.m.; Pb - 3 p.p.m; Ni - 5 p.p.m.


Histochemical reactions of gastrointestinal mucosubstances with orcein, high iron diamine and Alcian blue after prior oxidation of tissue sections. The histochemical orcein reaction (orc) for mucosubstances in tissue samples from the human gastrointestinal tract was compared with PAS, high iron diamine (HID) and Alcian blue reactions at pH 1.0 or 2.5 (AB 1 and AB 2.5). Orc, HID and AB 1 reactions were performed also with prior oxidation of the tissue sections with potassium permanganate or performic acid (ox-orc, ox-HID and ox-AB reactions, respectively). Orc reaction stained mucosubstances similarly to HID and AB 1; only the brush border and goblet cells in the colon were stained. The reactions of the mucosubstances obtained with ox-orc differed from those with PAS, HID, AB 1 or AB 2.5 but were similar to those with ox-HID or ox-AB; the mucosubstances in the brush border and the goblet cells in the colon and small bowel and in the foveolar epithelium of the stomach were strongly stained. Pyloric and cardiac glands were stained faintly with ox-orc but not with ox-HID or ox-AB. Brunner's glands were negative with ox-orc, ox-HID and ox-AB reactions. It was assumed that the orc reaction stains, like HID or AB 1, sulphate groups in epithelial mucosubstances, and that sulphonic acid residues, resulting from oxidation of disulphide groups in the protein core of mucus glycoproteins, are responsible for the ox-orc as well as for the ox-HID and ox-AB reactions.


Histochemical demonstration of a circadian rhythm of succinate dehydrogenase in rat pineal gland. Influence of coenzyme Q10 addition. Succinate dehydrogenase activity was investigated histochemically in the rat pineal gland. The influence of fixation on the activity pattern, the possible diffusion of enzyme, the nothing dehydrogenase reaction, and the substantivity of the tetrazolium salts and formazans were investigated in control experiments. In rats maintained on a 17/7 h light/dark schedule a distinct circadian rhythm of the succinate dehydrogenase was demonstrated in the pineal gland. Activity was lowest during the day and highest during the night. The dorsocaudal part of the gland showed the highest activity and within the same part of the gland the activity varied between individual pinealocytes. A relative lack of endogenous coenzyme Q, as well as a circadian rhythm of this coenzyme, highly influenced the activity of succinate dehydrogenase. It is concluded that succinate dehydrogenase activity in the pineal gland of the rat is regulated by changing the concentration of the active enzyme itself as well as the level of the endogenous coenzyme Q. Whether this is caused by a circadian rhythm in the synthesis or in the catabolism of the enzyme and the coenzyme was not revealed by the present study .


Histochemical and atomic absorption demonstration of trace metal mobilization in the central nervous system and liver of the rat. Histochemical and atomic absorption spectrophotometric analysis of trace metal mobilization caused by the action of ethanol in the central nervous system (CNS) and liver of the rat is described. Histochemically it has been shown that in all neurons examined (motoneurons, pyramidal and Purkinje cells) the trace metals (mainly Zn2+ and Cu2+) are mobilized. Most of the stained materials disappear from the perikaryon of the Purkinje cells, while in both the motoneurons and the pyramidal cells the trace metals are displaced from the perikaryon into the axon and axon hillock. At the same time, some of the glia cells display a high metal content. Quantitative determination of the Zn2+ and Cu2+ by means of atomic absorption spectrophotometry reveals that after 2 hours ethanol treatment both the Zn2+ and the Cu2+ levels are decreased in the archicerebellar cortex, while after 4 hours the Zn2+ levels are increased in the cerebrum and the spinal cord. The present observations on the histochemical localization and the contents of zinc and copper in different parts of the CNS and liver reveal the important role of the effect of ethanol on the trace metal mobilization.


Activities of DNA polymerases and RNA polymerases detected in situ in growing and differentiating cells of root cortex. Activities of DNA polymerases and RNA polymerases were studied by autoradiographic methods in growing and differentiating root cortex cells of Zea mays - a species in which endomitosis occurs - and Tulipa kaufmanniana - in which this process does not occur. In Tulipa kaufmanniana, the highest activity of DNA polymerase appears in the nuclei of meristematic zone during the S phase of the cell cycle. In Zea mays, endomitotic replication of DNA occurs in all growth and differentiation zones and the activity of DNA polymerase in the nuclei is similar to that in the meristematic zone. In both species, nuclear RNA synthesis, measured with 3H uridine incorporation, is highest in the meristematic zone and declines steadily with development. Activity of nuclear RNA polymerase is present in all developmental zones in both species and is similar to that in the meristematic zone. 3H uridine incorporation into nucleoli decreases markedly in both species, whereas the activity of nucleolar RNA polymerase remains at a high level in all root segments in Zea mays and decreases slightly in Tulipa kaufmanniana. It is argued that the differences between the incorporation of 3H uridine and that or 3H UMP may be caused by a reduction of the pool of endogenous ribonucleoside triphosphates. Marked activities of DNA polymerase and RNA polymerase in cytoplasm are possibly related to the growth and division of plastids and mitochondria.


Studies on the molecular arrangement of RNA in tissues with a selective topo-optical reaction of RNA. Selective demonstration of RNA in tissues was achieved by treating tissue sections with potassium permanganate followed by bisulfite and toluidine blue at pH 1.0 (PBT reaction). It is suggested that this reaction is due to aldehyde groups which are formed by the oxidative cleavage of the pyrimidine rings of RNA which can be selectively demonstrated using bisulfite-toluidine blue as the aldehyde reagent. The specificity of the reaction was tested after RNAase treatment, after acid hydrolysis, and on pure RNA droplets. The aldehyde nature of the reacting groups was checked, after permanganate oxidation, by Schiff's leucofuchsin reagent, and by aldehyde blocking reactions. Two types of intracellular molecular arrangement of RNA molecules could be distinguished by polarization optics after application of the PBT reaction: 1) The strong birefringence, dichroism and metachromatic staining of membrane-bound RNA in ergastoplasm of pancreas, liver and plasma cells indicate a linear (planar) molecular order of RNA molecules on the surface of the membranes, and 2) the isotropic, basophilic staining of RNA not organized in membrane structures (Nissl substance, nucleoli) suggest a random distribution of their dye binding sites.


Combination of immunological and lectin reactions in affinity histochemistry: proposition of the term affinitin. Using the series system cell receptor leads to mistletoe lectin leads to antiferritin-antibody leads to ferritin, the possibilities for combination of lectin and immunological reactions for histochemistry are discussed. The system cell antigen leads to antibody leads to labelled mistletoe (or other) lectin is recommended for visualization of cell antigens (mistletoe lectin as common immunoglobulin reagent). It is pointed out that lectin reactions do not belong to immunhistochemistry but to affinity histochemistry. For all receptor specific proteins (antibodies, lectins, enzymes, haptoglobin and other) the term affinitin is proposed. In consideration of this new definition a common scheme is formulated: Affinitin reacts with affinitin receptor forming affinity product.


[Demonstration of sulphur in secretory granules of anterior and posterior pituitary by X-ray microanalysis (author's transl)]. The sulphur content of secretory granules and other subcellular structures in the cells of the anterior pituitary and of neurosecrotory granules in the nerve fibers of the neural lobe have been studied by energy dispersive microanalysis. The highest level has been found in the secretory granules of the neural lobe.


Supercontraction in crayfish muscle: correlation with a peculiar actin localization. Crayfish muscle, like muscles from some other invertebrates, can supercontract. This muscle shortening is characterized by an overlap of thin filaments with crossing of thick filaments through the Z discs. In intact muscle cells, supercontraction does not seem to induce irreversible structural modifications in the tissue. Isolated crayfish myofibrils in the relaxed state cannot be distinguished from vertebrate myofibrils under light microscope, either by phase contrast or by immunofluorescence, with antiactin antibodies, actin being localized in the I bands. However, when isolated crayfish myofibrils are supercontracted, irreversible dammage occurs, most thin filaments being lost. Actin becomes then hardly detectable, being visible, by immunofluorescence, either in the Z discs or evenly distributed in the whole myofibril. During myofibril supercontraction, high amounts of denatured actin, become soluble as shown by SDS-PAGE, by double immunodiffusion, and by DNAse inhibition.


NADP-dependent dehydrogenases in rat liver parenchyma. II. Comparison of qualitative and quantitative G6PDH distribution patterns with particular reference to sex differences. Qualitative histochemical G6PDH distribution patterns obtained in the liver acinus of adult male and female rats with an improved method (Rieder et al., 1978) served as a basis for the isolation by microdissection of tissue samples of defined zonal affiliation. G6PDH activity was assayed quantitatively in tissue samples of zones 1 and 3 by a microfluorometric method, using the oil well technique and enzymatic cycling (Burch et al., 1963; Lowry and Passonneau, 1972). With the use of a correlation system further evidence could be presented for the validity of the recently described qualitative distribution patterns. From a total of 50 analyzed tissue samples the following G6PDH activities were calculated: 4.25 +/- 1.56 U/g dry weight in zone 1 and 2.08 +/- 0.46 U/g dry weight in zone 3 of male and 7.21 +/- 1.03 U/g dry weight in zone 1 and 11.10 +/- 2.56 U/g dry weight in zone 3 of female rats. These data were corrected for interference from the G6PDH activity of the Kupffer cells within zone 1 samples (approximately 80 U/g dry weight), so that the actual relative values for the parenchymal activity could be estimated for the first time: 2 U/g dry weight in zones 1 and 3 of male animals, 5 U/g dry weight in zone 1 and 11 U/g dry weight in zone 3 of female animals. In female livers G6PDH activity in zone 1 is therefore 2.5 times higher, and in zone 3 5 times higher than in the male. These zonal as well as sex-differences are clearly indicative of a heterogeneous functional organization of the liver acinus in terms of capacity for NADPH production, mainly in connection with reductive reactions in fatty acid synthesis.


Principles and methods for the quantitative determination of Feulegen stained DNA with the television texture analysis system (TAS). The television texture analysis system (TAS, Leitz) has been applied to determination of the relative DNA content of Feulgen stained nuclei. The principles of the method are described and comparative measurements are presented. The determination is based on the measurement of areas at preset intervals of transmittance, taking into consideration the necessary correction of extinction values in relation to area and applying a correction for background. The data obtained can be used for the quantitative determination of DNA, and to provide analytical information for quantitation of structure. Comparison of the mean of extinctions as recorded with the scanning photometric method and those recorded with the television texture analysis method, with which 43 nuclei were investigated in transmittance steps of 0.01, revealed a coefficient of correlation of 0.9938.


Histochemical demonstration of alkaline phosphatase in human large intestine, normal and diseased. Alkaline phosphatase activity has been investigated by histochemical methods in normal and diseased human large intestine. The tissues were constantly maintained at 4 degrees C or below. Specimens were either frozen in liquid nitrogen, freeze-dried and embedded in glycol methacrylate for sectioning at 2 mu, or, fixed in ice-cold formol-calcium for frozen sectioning at 10 mu. The simultaneous coupling azo dye method using the substrates sodium alpha-naphthyl phosphate and Naphthol AS-BI phosphate, resulted in the demonstration of alkaline phosphatase activity in the surface epithelial cells, and the middle and upper crypts, of normal and transitional mucosa.


[Automated body plethysmography of respiration and voice efficiency (author's transl)]. Efficiency of voice (EV) is understood as the ratio of gained acoustic power to respiratory power during phonation. A fully automatic computer programmed to calculate the EV and the subglottic pressures via body plethysmography is described. The EV ranges from 0.5 x 10(-5) to 120 x 10(-5). The relationship between EV and sound pressure is demonstrated. The results were reproducable.


[Aquamat--a proven bathing aid for the laryngectomee (author's transl)]. A device to facilitate bathing by the laryngectomee is described. The Aquamat is made up of equipment routinely used in anaesthesia and underwater diving. An airway link is made between a watertight tracheostoma and the mouth such that the patient can breath between nose and trachea. Special precautions are necessary for swimming.


[Therapy of sudden deafness with O2/CO2 inhalation (author's transl)]. A total of 55 patients with sudden deafness have been treated in a prospective randomized study by inhalation with 95% O2 and 5% CO2 or intravenous infusion with papaverin and dextran. Five days after treatment, no significant difference could be found between both therapies. However, audiometric results obtained one year later show statistically better results for those patients who were managed by O2/CO2 inhalations and indicate that such therapy may improve the spontaneous rate of recovery inherent to sudden deafness.


[Common boundaries of otorhinolaryngology and ophthalmology (author's transl)]. The necessity for close co-operation between otorhinolaryngologist and ophthalmologist is demonstrated with clinical examples. Surgical approaches familiar to the otolaryngologist offer greater opportunity for optimal patient care particularly in the management of orbital tumours and trauma.


[Vertigo and vertebral artery stenosis (author's transl)]. Forty nine elderly patients with osteochondrosis of the cervical spine and on whom carotid angiography had been performed were divided into two groups. In one 36 patients had vestibular symptoms whereas the other group of remaining patients was free of such symptoms. Comparison of both groups showed that vertigo (20 patients), other vestibular symptoms or nystagmus were not solely related to vertebral artery stenosis. In fact stenosis of one or both vertebral arteries was found in only nine patients.


Clinical and radiographic signs in primary and metastatic esophageal neoplasms of the dog. Esophageal neoplasms were diagnosed in 8 of 49,229 dogs seen over the last 11 years at the Veterinary Medical Teaching Hospital of the University of California at Davis. The neoplasms were primary in 2 dogs and metastatic to the esophagus in 6 dogs, with thyroid carcinoma being the most common site of the primary tumor. The most common clinical signs were regurgitation, dysphagia, weight loss, development of neck masses, and respiratory difficulties. It was concluded that the clinical signs often can be misleading. The interpretation of survey radiographs, barium contrast studies, or fluoroscopic studies often provide the initial data base. The final diagnosis requires histologic examination. Retention of air in the esophagus (with or without esophageal displacement) and motor dysfunction (with or without gross morphologic changes) are the most important criteria for radiographic diagnosis.


Urinary tract infection induced by intermittent urethral catheterization in dogs. Sterilized and unsterilized catheters were passed into the urinary bladders of 9 clinically normal adult male dogs once daily for 5 consecutive days, and the dogs were examined for up to 30 days to determine whether urinary tract infections developed. Two dogs that were catheterized with clean unsterilized catheters (1 clinically normal dog and 1 dog given immunosuppressant drugs) developed persistent cystitis and pyelonephritis due to infection with Proteus sp. One dog given immunosuppressant drugs developed a mixed bacterial infection (Proteus sp and Escherichia coli) that resolved without treatment between 22 and 30 days later.


Canine hypertrophic cardiomyopathy.

Canine hypertrophic cardiomyopathy. Necropsy findings in 10 dogs with naturally occurring cardiac disease closely resembled hypertrophic cardiomyopathy in human beings and cats. Each dog had marked cardiac hypertrophy, and 8 dogs had disproportionate thickening of the ventricular septum with respect to the left ventricular free wall (compared with dogs with normal hearts or with cardiac hypertrophy due to acquired or congenital heart disease). Ratios of septum to free wall thickness in the 10 dogs ranged from 1.1 to 1.5, and 6 had ratios greater than or equal to 1.3. Marked cardiac muscle cell disorganization in the ventricular septum, characteristic of human patients with hypertrophic cardiomyopathy, was found in only 2 of the 10 dogs. Death occurred while the dogs were under anesthesia during the course of operative procedures (5 dogs) or unexpectedly in animals without previous manifestations of cardiac disease (3 dogs). Four dogs had clinical signs of congestive heart failure, including 2 with marked cardiac decompensation. Two of these 4 dogs with heart failure and 1 dog that died during unrelated surgery, but without prior signs of heart disease, had electrocardiographic evidence of complete heart block.


Dermatitis associated with Cheyletiella infestation in cats. Cheyletiella sp mites were isolated from 8 cats with pruritic dermatitis characterized by erythematous papules or increased scaling. Lesions cleared after treatments with malathion (dips) or pyrethrin (shampoos).


Adenocarcinoma of the frontal sinus with extension to the brain in a horse. A space-occupying intracranial mass was diagnosed in a horse. The clinical findings included blindness, circling to the right, apprehension, anorexia, weight loss, and leaning against the stall. On ophthalmoscopic examination, the most striking observation was complete bilateral devascularization of the retinas. The horse was euthanatized and necropsied. Necroscopy revealed the mass to occupy the olfactory and frontal areas of the left hemisphere of the brain and part of the left frontal sinus. Microscopically, the mass was an adenocarcinoma and probably arose from the lining epithelium of the sinus or from the subepithelial glands.


Decreased tear production associated with general anesthesia in the dog. Schirmer I tear tests were done on 20 dogs before and during general anesthesia. Atropine sulfate reduced tear production from a mean base-line value of 20.35 +/- 3.33 mm/minute to 14.10 +/- 6.13 mm/minute, 10 minutes after its subcutaneous administration. The combination of subcutaneous atropine, intravenous thiamylal sodium, and inhalation anesthesia reduced tear production to a mean value of 0.53 +/- 1.12 mm/minute at 60 minutes after induction.


Comparative studies on actins from various sources. Fragments of actins from Ascaris muscle cleaved at cysteinyl residues in comparison with those of other actins. Pure actins were obtained from various animal muscles: Vertebrata (skeletal, smooth, and cardiac muscles), Prochordata (smooth muscle), Nematoda (obliquely striated muscle), and Mollusca (striated, smooth and obliquely striated muscles). These actins were all identical in apparent molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. All actins treated with 2-nitro-5-thiocyanobenzoic acid yielded four major (about 33,000, 26,000, 24,000, and less than 10,000 daltons) and three minor (22,000, 17,000, and 10,000 daltons) bands in addition to intact actin on gel electrophoresis. The results suggest that all actins from various types of muscle have cysteinyl residues at similar positions on the primary structure.


Properties of unprimed poly(A)-poly(U) synthesis by Caulobacter crescentus RNA polymerase. Some properties of unprimed poly(A)-poly(U) synthesis by DNA-dependent RNA polymerase from Caulobacter crescentus were examined. The reaction required ATP and UTP as substrates and manganese as a divalent cation. Rifampicin completely inhibited the reaction at a concentration of 1 micron/ml, and the enzyme catalyzed the polymer synthesis well regardless of the presence of GTP, CTP or both. The chain length of the poly(A)-poly(U) synthesized was about one hundred base pairs, as estimated from a sedimentation velocity and the molar ratio of [3H]AMP to [gamma-32P]ATP incorporated into the poly(A)-poly(U). The reaction was dependent on the square of the enzyme concentration and the enzyme dimers formed complexes with poly(A)-poly(U) during the reaction.


Purification and properties of cytochrome b-562.5 from Ulva pertusa. Cytochrome b-562.5 (Ulva pertusa) was extracted from a green alga, U. pertusa, by homogenization of the thalli in phosphate buffer solution. Purification was carried out by acrinol treatment, ammonium sulfate fractionation, DEAE-cellulose and DEAE-Sephadex column chromatographies, and Sephadex gel filtration. Cytochrome b-562.5 has absorption maxima at 562.5  (alpha), 530.5 (beta), 429 (gamma), and 326 nm (delta) in the reduced form and at 537, 415 (gamma), and 275 nm in the oxidized form. The alpha-band of the reduced form is asymmetric with a shoulder at 560 nm, at liquid nitrogen temperature this band splits into two distinct peaks at 562 and 556.5 nm. The absorption maxima of the pyridine ferrohemochrome appear at 556 (alpha), 523 (beta), and 418 nm (gamma). The cytochrome does not combine with carbon monoxide or cyanide. The preparation of the cytochrome shows little peroxidase activity. The cytochrome is oxidized by ferricyanide and reduced by cysteine, ascorbate, and hydrosulfite. Autoxidation of the cytochrome was found to be very slow. The midpoint potential (Em) of the cytochrome was determined by equilibration with the ferro- and ferri-EDTA system to be +0.20 V at pH7.0. The molecular weight of the cytochrome was estimated by Sephadex gel filtration to be 23x10(3).


Structures of the asparagine-linked sugar chains of human chorionic gonadotropin. The asparagine-linked sugar chains of human chorionic gonadotropin were released from the polypeptide moiety by hydrazinolysis followed by N-acetylation and NaB3H4 reduction. More than 90% of the released radioactive oligosaccharides contained N-acetylneuraminic acid residues. After removal of N-acetylneuraminic acid residues by sialidase treatment, two neutral oligosaccharide fractions were obtained by paper chromatography. Sequential exoglycosidase digestion revealed that one of them was a mixture of two neutral oligosaccharides. The complete structures of the three oligosaccharides were elucidated by methylation analysis. It was confirmed that all the N-acetylneuraminic acid residues of the asparagine-linked sugar chains of human chorionic gonadotropin occur as NeuAc alpha 2 leads to 3Gal groupings by comparing the methylation analysis data for the acidic oligosaccharide mixture before and after sialidase treatment. Based on these results, the structures of the asparagine-linked sugar chains of human chorionic gonadotropin were confirmed to be +/- NeuAc alpha 2 leads to 3Gal beta 1 leads to 4GlcNAc beta 1 leads to 2Man alpha 1 leads to 6(NeuAc alpha 2 leads to 3Gal beta 1 leads to 4GlcNAc beta 1 leads to 2Man alpha 1 leads to 3)Man beta 1 leads to 4GlcNAc beta 1 leads to 4(+/- Fuc alpha 1 leads to 6)GlcNAc and Man alpha 1 leads to 6(NeuAc alpha 2 leads to 3 Gal beta 1 leads to 4 GlcNAc beta 1 leads to Man alpha 1 leads to 3)Man beta 1 leads to 4 GlcNAc beta 1 leads to 4GlcNAc.


Effect of dietary proteins on the turnover of rat liver argininosuccinate synthetase. The rates of synthesis and degradation of arginosuccinate synthetase in rat liver under various dietary conditions were determined. The relative rate of the enzyme synthesis in the livers of rats fed on 70% casein diet was 4.0 times greater than that for rats fed on 5% casein diet. The rate constants of degradation (Kd of argininosuccinate synthetase were estimated to be 0.15 and 0.16 day-1 under 70% and 5% casein feeding, respectively. When the dietary conditions were changed acutely from 70% to 5% casein diet or vice versa, the rates of the enzyme synthesis decreased or increased, respectively, and the rates of enzyme degradation were also affected. The change from 5% to 70% casein diet caused a transient decrease in the rate of degradation. After the enzyme activity had achieved a new steady-state level, the enzyme degradation proceeded at the normal steady rate. On the other hand, the change from 70% to 5% casein diet caused a transient increase in the rate of degradation. Thus, the only factor regulating the amount of enzyme in rat liver is the rate of enzyme synthesis under the steady-state conditions. However, the rates of both enzyme synthesis and degradation are involved in the regulation of the amount of enzyme during dietary transition.


Biochemical studies on sulfate-reducing bacteria. XV. Separation and comparison of two forms of desulfoviridin. Desulfoviridin from Desulfovibrio vulgaris was separated into two forms by DEAE-Sephadex column chromatography. The major form had a pI of 4.4 and the minor form one of 4.5-4.6. Both forms produced mainly trithionate, besides thiosulfate and sulfide, in methylviologen-linked sulfite reduction. The specific activities of sulfite reduction, as well as of hydroxylamine reduction, were virtually identical in both forms. There were no great differences in their absorption spectra, CD spectra, molecular weights, subunit compositions, labile sulfide, and iron contents, and amino acid compositions. The N-terminal amino acid was alanine in both forms.


Effect of nucleoside 5'-triphosphates on tRNA nucleotidyltransferase activity in cytoplasmic fractions of various types of mammalian cells. The tRNA nucleotidyltransferase activity (3H-CMP incorporation into 3'-terminus of tRNApC) in cytoplasmic fractions of various types of cells such as Ehrlich ascites tumor cells, mouse liver and spleen cells, rat spleen, lymph node, and macrophages cells was found to be dependent on the concentrations of nucleoside 5'-triphosphates (ATP, GTP, UTP, dATP, dGTP, dCTP, and/or dTTP). The purified tRNA nucleotidyltransferase did not show such dependency. The dependency of the enzyme activity on nucleoside 5'triphosphates in the crude cytoplasmic fractions was possibly due to the presence of inhibitors which interfere with the repair system of defective 3'-termini of tRNA. Two kinds of inhibitors were distinguishable in the cytoplasmic fractions. One was unstable on heat treatment at 55 decrees C and showed ribonuclease activity for the tRNA 3'-terminus. The other which lacked ribonuclease activity was rather stable to the heat treatment and inhibited purified tRNA nucleotidyltransferase. The actions of both inhibitors were suppressed by nucleoside 5'-triphosphates.


The mechanism of decrease in nucleolar RNA synthesis by protein synthesis inhibition. When protein biosynthesis is inhibited by either cycloheximide of puromycine, the nucleolar RNA synthesis of Ehrlich ascites tumor cells decreases by approximately 70% within 1 h, while the removal of these protein synthesis inhibitors causes a rapid recovery of nucleolar RNA synthesis, largely within 1 h. A similar pattern of decrease and recovery of endogenous RNA polymerase activity in isolated nucleoli or in nuclei (in the presence of alpha-amanitin) may be demonstrated after addition and removal of these drugs. Analysis of the molecular species of RNA polymerase I on a phosphocellulose column indicates that only the IB form of the enzyme decreases in the nucleoli of drug-treated cells and recovers quickly after resumption of protein synthesis. The finding that the activity of the IB form enzyme remains unchanged in the whole nuclei indicates that during cessation of protein synthesis RNA polymerase IB is either released from the nucleoli into the extranucleolar compartment or becomes so loosely bound to the nucleoli that it is leached out from the nucleoli during their isolation. By using a system of assaying free, nucleolar-template bound and total RNA polymerase I activities, data supporting the above interpretation have been obtained. Namely, in isolated nuclei free enzyme activity increases with a concomitant decrease in bound enzyme activity during protein synthesis inhibition, while the total enzyme activity remains unchanged. In isolated nucleoli, both total and bound enzyme activities decreases on protein synthesis inhibition but recover quickly on its resumption. The putative bound enzyme, fractionated with the aid of actinomycin D, is exclusively IB form, whereas the unbound enzyme consists of both IA and IB forms as previously demonstrated (1). No conversion of IB form polymerase to IA form was noted on prolonged sonication in our system. The levels of ATP and GTP in the cell did not change appreciably either during cessation or resumption of protein synthesis in these cells. The data support the previous conclusion that some short-lived protein(s) is required to maintain the normal level of ribosomal RNA transcription (2) and further suggest that the protein is required to facilitate reinitiation of the transcription by RNA polymerase IB in the nucleolus.


Stability mapping along the DNA double strand and its relation to the genetic map. The distribution of the stability of the double helical structure along the whole DNA of fdphage and its restriction fragments is calculated. In this calculation, Poland's method, which has been established as a rigorous algorithm for taking the base sequence explicitly into consideration, is used. The molecular thermodynamic parameters in the calculation have been determined so as to best reproduce the melting profile of the DNA and its fragments. The results, which are presented as melting maps, show fairly good agreement with those experimentally obtained by the present authors earlier. A close correlation with genes in the genetic map is apparent for some cooperatively melting regions observed in the stability map.


Thermal regulation of fatty acid synthetase from Brevibacterium ammoniagenes. The fatty acid composition of Brevibacterium ammoniagenes was affected by the temperature of growth. As the growth temperature was lowered, the proportion of unsaturated fatty acids increased. The fatty acid synthetase obtained from B. ammoniagense produced oleic acid as well as saturated fatty acids. The ratio of unsaturated to saturated fatty acids synthesized by this enzyme in vitro was dependent on the temperature of the enzyme reaction but not on the growth temperature of B. ammoniagenes from which the enzyme was prepared. These results suggest that the changes of composition in cellular fatty acids reflect the temperature dependence of the fatty acid synthetase.


Preparative use of the analytical column of a sugar autoanalyzer for resolution of gluco-oligosaccharides of the same molecular weight. A sugar autoanalyzer was used on a preparative scale to resolve a gluco-oligosaccharide mixture. In this way the components of the following mixtures were resolved: O-alpha-D-glucopyranosyl-(1-3)-O-[alpha-D-glucopyranosyl-(1-6)]-D-glucose (1), O-alpha-D-glucopyranosyl-(1-6)-O-alpha-D-glucopyranosyl-(1-3)-D-glucose (2) and O-alpha-D-glucopyranosyl-(1-3)-O-alpha-D-glucopyranosyl-(1-6)-D-glucose (3), O-alpha-D-glucopyranosyl-(1-3)-O-alpha-D-glucopyranosyl-(1-4)-D-glucose (4) and O-alpha-D-glucopyranosyl-(1-4)-O-alpha-D-glucopyranosyl-(1-3)-D-glucose (5), and O-alpha-D-glucopyranosyl-(1-2)-O-alpha-D-glucopyranosyl-(1-6)-O-alpha-D-glucopranosyl-(1-6)-O-alpha-D-glucopyranosyl-(1-6)-D-glucose (6) and O-alpha-D-glucopyranosyl-(1-3)--O-alpha-D-glucopyranosyl-(1-6)-O-alpha-D-glucopyranosyl-(1-6)-O-alpha-D-glucopyranosyl-(1-6)-D-glucose (7).


Drafting film in medical illustration.

Drafting film in medical illustration. The use of drafting film in creating pen and ink medical illustrations is generally superior to scratch board. Effects of scratch board may be duplicated with drafting film. In addition, the cost per page is lower, transfer time is saved, and the film is completely erasable without damage to the working surface.


The case history of a continuing medical education film. The key provisions necessary in the development, implementation, and evaluation of a film designed for Continuing Medical Education are described. A case history approach is utilized with discussion on each of the following points: (1) problem identification, (2) audience, (3) media selection, (4) media production, (5) research design, (6) media distribution, (7) evaluation and (8) results. Attention to each of the above items contributed to this project's success, and reaffirms the important role which the medium of film can have in continuing professional education.


Fluorescent photochemical surface labeling of intact human erythrocytes. A photolabile nitrene precursor, 3-azido-(2,7)-naphthalene disulfonate (ANDS), has been synthesized and used as a membrane-impermeable probe. The aryl azide was nonfluorescent. When activated by light, a highly reactive nitrene was generated which was capable of nonspecific covalent modifications of hydrophilic regions of cell surfaces. The products of the photolysis were highly fluorescent and modified proteins could be identified by their characteristic fluorescence after electrophoresis on sodium dodecyl sulfate polyacrylamide gels. When intact human erythrocytes were labeled with ANDS, Protein 3, the major membrane protein, and the sialoglycoproteins were modified. No proteins of apparent molecular weight greater than Protein 3 were labeled by ANDS, suggesting that none of these membrane components was exposed to the hydrophilic external surface of the red blood cell. When open erythrocyte stroma were labeled with ANDS, virtually all protein bands detectable by Coomassie blue staining could be shown to contain some fluorescence label. The significance of these findings are discussed with relation to the use of various aryl azides as surface labels of membranes.


Affinity isolation and characterization of immunoglobulin G Fc fragment-binding glycoprotein from human blood platelets. Immune complexes bind to several eukaryotic cell types including human blood platelets through the Fc fragment of immunoglobulin (Ig) G. Utilizing immobilized Fc fragment of IgG enabled us to isolate from human blood platelets a glycoprotein of an apparent Mr = 255,000 which, upon reduction, dissociated into sub-units of an apparent Mr = 50,000. This Fc fragment-binding glycoprotein has an isoelectric point between pH 6.3 and 6.9 and is composed of 34% hydrophobic, 25% acidic, and 14% basic amino acids. The Fc fragment-binding glycoprotein was also isolated from human platelet membrane preparations and was unaffected by prior treatment of platelets with thrombin. Isolated Fc fragment-binding glycoprotein formed an in vitro complex with aggregated immunoglobulin G. These results suggest that the isolated Fc fragment-binding component may prove useful in studies concerning the functional role of glycoproteins as cellular receptors for the Fc fragment of IgG.


The structure and amount of tubulin in cells and tissues. The NH2-terminals and amino acid compositions of three carboxymethylcysteine-containing peptides produced from carboxymethylated calf brain tubulin by digestion with elastase have been determined. The 3H/14C ratio obtained when these peptides were generated from mixtures of [14C]carboxymethylated calf brain tubulin and the [3H]carboxymethylated proteins of mouse brain or 3T3 cells and subsequently purified indicated that both mouse brain and 3T3 cells contained protein giving rise to peptides homologous to these peptides. This indicates that the tubulins of calf brain, mouse brain, and 3T3 cells are homologous in the regions of amino acid sequence accounting for the peptides examined. From the 3H/14C ratio of the isolated peptides, 13.5% of the protein of mouse brain and 3.5% of the protein of 3T3 cells were estimated to be tubulin.


An actin-like protein is a component of axonemes from Chlamydomonas flagella. Several lines of evidence indicate that a component of axonemes from Chlamydomonas flagella is similar to actin from rabbit skeletal muscle. The polypeptide has an apparent molecular weight of 42,000 and in a pH gradient has the electrophoretic behavior of beta-actin. It was co-polymerized with rabbit actin and purified by affinity chromatography on DNase I-Sepharose. Incomplete proteolysis of mixed 35S-labeled axonemal protein and cold rabbit actin formed similar sets of peptides as analyzed by one-dimensional gel electrophoresis. The actin-like protein and the tubulin appear to be present in the axoneme in the molar ratio 1:60.


Phosphorylation of paramyosin and its possible role in the catch mechanism. Paramyosin isolated from the adductor muscle of Mercenaria mercenaria was shown to contain three to five phosphate groups/molecule; the actual number varied depending on the method used to extract the protein. Dephosphorylation resulted in an increase in the solubility of paramyosin near pH 7 and near physiological ionic strength. This behavior suggests that the number of phosphates/molecule may be a determining factor in the aggregation behavior of paramyosin-containing myofilaments. Thus, phosphorylation may be involved in catch contractions since correlations have been demonstrated earlier between catch contraction of molluscan muscles and aggregation properties of paramyosin (Ruegg, J. C. (1971) Physiol. Rev. 51, 201-249).


Iodide binding and regulation of lactoperoxidase activity toward thyroid goitrogens. The effects of the antithyroid goitrogens, methylthiouracil and methylmercaptoimidazole, on the oxidation of N-acetyltyrosylamide at pH 8.8 by lactoperoxidase have been evaluated in the presence and the absence of iodide for the purpose of elucidating the effects of iodide. At pH 8.8, iodine is not oxidized. In the absence of iodide, the two antithyroid drugs inactivate lactoperoxidase by a second order process. When iodide is added before methylthiouracil or methylmercaptoimidazole, enzyme inactivation does not occur as rapidly and both goitrogens are readily oxidized. The kinetics of the oxidation reactions have been analyzed in order to obtain the equilibrium constant of the iodide . lactoperoxidase complex. Essentially the same iodide dissociation constant, i.e. 2 x 10(-5) M, was found by studying its effects on the kinetics of oxidation of the two antithyroid drugs. A large difference absorption spectrum is observed in the Soret region between native lactoperoxidase and lactoperoxidase inactivated by methylthiouracil.


Insulin-stimulated intracellular hydrogen peroxide production in rat epididymal fat cells. Insulin stimulation of hydrogen peroxide production by rat epididymal fat cells was investigated by studying the oxidation of formate to CO2 by endogenous catalase. Under optimal concentrations of formate (0.1 to 1 mM) and glucose (0.275 mM), insulin stimulated formate oxidation 1.5- to 2.0-fold. Inhibitors of catalase activity, including nitrite and azide, inhibited both basal and insulin-stimulated formate oxidation at concentrations that did not interfere with insulin effects on glucose C-1 oxidation or glucose H-3 incorporation into lipids. The addition of exogenous catalase increased formate oxidation only slightly, while exogenous H2O2 (0.5 mM) stimulated formate oxidation by endogenous catalase strongly. These data indicate that the insulin-stimulated H2O2 production was intracellular. Insulin dose-response curves for formate oxidation were identical with those for glucose H-3 incorporation into lipids. The dependence of relative insulin effects on the logarithm of the glucose concentration was bell-shaped for formate oxidation and correlated highly with the coresponding dependences of glucose C-1 oxidation and glucose H-3 incorporation into lipids. This suggests that insulin stimulation of intracellular H2O2 production is linked to glucose metabolism. Since it is known that extracellular H2O2 can mimic insulin in several respects, these observations suggest that H2O2 may act as a "second messenger" for the observed effects of insulin.


Carbon 13 NMR spectroscopy of a cerebroside. Proof of the beta-pyranosyl structure of D-glucosylceramide. The first 13C nuclear magnetic resonance spectrum of a cerebroside is reported. All signals of D-glucosylceramide (I), the cerebroside from human Gaucher's diseased liver, have been assigned by comparison of their chemical shifts with those of model compounds. This data has been used to demonstrate for the first time directly: (a) that the glucosyl moiety must exist in the pyranoside ring form and have a beta anomeric linkage, and (b) that the fatty acyl composition of the aglycon is: alpha-hydroxy, 0%; saturated, 86 +/- 2%; unsaturated long chain, 14 +/- 2%; and unsaturated short chain, less than or equal to 2%. The use of 13C signal shifts in the delta 80 to 120 region (anomeric window) for the assignment of glycon anomeric and ring forms demonstrated herein should be applicable to other glycoconjugates.


Purine and pyrimidine transport and phosphoribosylation and their interaction in overall uptake by cultured mammalian cells. A re-evaluation. The zero-trans uptake of purines and pyrimidines was measured in suspensions of Novikoff rat hepatoma, mouse L, P388 mouse leukemia, and Chinese hamster ovary cells by a rapid kinetic technique which allows the determination of uptake time points in intervals as short as 1.5 s. Kinetic parameters for purine/pyrimidine transport were determined by measuring substrate influx into cells in which substrate conversion to nucleotides was negligible either due to lack of the appropriate enzymes or to depletion of the cells of ATP (5'-phosphoribosylpyrophosphate), and by computer fitting exact, integrated rate equations derived for various carrier-mediated transport models directly to zero-trans influx data. The results indicate that different carriers function in the transport of hypoxanthine/guanine, adenine, and uracil with substrate:carrier association constants (K) at 24 degrees C of 300 to 400 muM, 2 to 3 mM, and about 14 mM, respectively, for Novikoff cells. K and Vmax for hypoxanthine transport by L and P388 cells are similar to those for Novikoff cells, but the transport capacity of Chinese hamster ovary cells is much lower and K = 1500 muM. All transport systems are completely symmetrical. Hypoxanthine transport is so rapid that an intracellular concentration of free hypoxanthine (90%) close to that in the medium is attained within 20 to 50 s of incubation at 24 degrees C, at least at extracellular concentrations below K. In cells in which conversion to nucleotides is not blocked free hypoxanthine accumulates intracellularly to steady state levels with equal rapidity and thereafter the rate of hypoxanthine uptake into total cell material is strictly a function of the rate of phosphoribosylation. The low Km systems for hypoxanthine (1 to 9 muM) and adenine (0.2 to 40 muM) uptake detected previously in many types of cells reflect the substrate saturation of the respective phosphoribosyltransferases rather than of the transport system.


Bilirubin. Solubility and interaction with albumin and phospholipid. Bilirubin is generally considered a lipophilic substance, and its neurotoxicity is ascribed to an affinity for lipids in the central nervous system. In the present paper, it is shown that the solubility of bilirubin in apolar solvents and in triglycerides is low and increases with solvent polarity. Consequently, bilirubin should not be characterized as lipophilic. The solubility in aqueous buffers was studied under exclusion of light and was found lower than previously reported, about 7 nM at pH 7.4, temperature 37 degrees C, increasing with higher pH, approximately in inverse proportion with the squared hydrogen ion concentration. Binding of bilirubin to human serum albumin was studied by the rate of oxidation with peroxidase of the free ligand at equilibrium. The stoichiometry of proton involvement in the binding process was investigated by acidimetric titration. It is concluded that precipitation of bilirubin in vivo is thermodynamically possible. It was further demonstrated by light absorption spectroscopy that bilirubin forms a complex with phosphatidylcholine in diethyl ether and that an aqueous suspension of phosphatidylcholine enhanced aggregation of bilirubin. Transfer of bilirubin dianion from its complex with plasma albumin and precipitation of bilirubin acid with phosphatidylcholine in membranes of nerve cells is therefore possible and may account for the neurotoxicity.


The isolation and characterization of the link proteins from proteoglycan aggregates of bovine nasal cartilage. A preparation containing the link proteins may be obtained from bovine nasal cartilage by extraction with 4 M guanidine hydrochloride and by equilibrium density gradient centrifugations of the extract as commonly employed in the isolation of proteoglycan monomers. In the present paper, protein-rich proteoglycans have been removed from such a preparation to give purified link proteins by chromatography on Sepharose CL-6B in 1% sodium dodecyl sulfate. The individual link proteins, which in order of increasing electrophoretic mobility are termed link proteins 1, 2, and 3, have been separated and isolated in a subsequent preparative gel electrophoresis step. The link proteins present in largest amount, link proteins 1 and 2, have essentially the same amino acid compositions, and following partial digestion with the V8 protease from Staphylococcus aureus and analytical electrophoresis in sodium dodecyl sulfate, their peptide patterns closely resemble each other. Therefore,it is probable that link proteins 1 and 2 are structurally similar. Link protein 1 contains more carbohydrate than link protein 2 (9.5% and 3.0%, respectively) and it is suggested that the major difference between them is in carbohydrate content.


Substrate-dependent dissociation of malate thiokinase. Malate thiokinase has been purified to apparent homogeneity by employing conventional purification techniques along with affinity chromatography. The enzyme is composed of two nonidentical subunits (alpha subunit Mr=34,000, beta subunit Mr=42,500) to yield an alpha 4 beta 4 structure for the native enzyme. Phosphorylation of the enzyme by ATP occurs exclusively on the alpha subunit. The phosphorylated enzyme is acid labile and base stable consistent with phosphorylation of a histidine residue. Dephosphorylation of the enzyme is promoted by ADP, succinate, malate, and coenzyme A plus inorganic phosphate. Phosphorylation of the enzyme leads to a reversible change in the sedimentation properties of the enzyme; the native enzyme exhibits an S20,w of approximately 10, whereas the phosphoenzyme exhibits an S20,w of approximately 7. Formation of the 7 S form of the enzyme is also observed when coenzyme A and succinyl-CoA interact with the enzyme. The ratio of alpha to beta subunits in both the 10 S and 7 S forms of the enzyme is approximately 1.0, suggesting that the 7 S form of the enzyme has an alpha 2 beta 2 structure.


Binding and release of radiolabeled eukaryotic initiation factors 2 and 3 during 80 S initiation complex formation. The AUG-dependent formation of an 80 S ribosomal initiation complex was studied using purified rabbit reticulocyte initiation factors radiolabeled by reductive methylation. The radiolabeled initiation factors were as biologically active as untreated factors. Reaction mixtures containing a variety of components (AUG, GTP, Met-tRNAf, initiation factors, and 40 S and 60 S ribosomal subunits) were incubated at 30 degrees C and then analyzed on linear sucrose gradients for the formation of ribosomal complexes. The results show that both eukaryotic initiation factor (eIF)-3 and the ternary complex (eIF-2.GTP.Met-tRNAf) bind independently to the 40 S subunit and each of these components enhances the binding of the other. All of the polypeptides of eIF-2 and eIF-3 participate in this binding. Formation of an 80 S ribosomal complex requires eIF-5 and 60 S subunits in a reaction that is stimulated by eIF-4C. Both eIF-2 and eIF-3 are released from the 40 S preinitiation complex during formation of the 80 S initiation complex. Release of eIF-2 and eIF-3 does not occur and 80 S ribosomal complexes are not formed if GTP is replaced by a nonhydrolyzable analog such as guanosine 5'-O3-(1,2-mu-imido)triphosphate. Despite a variety of attempts, it has not yet been possible to demonstrate binding of eIF-4C, eIF-4D, or eIF-5 to either 40 S or 80 S ribosomal complexes.


Identification by peptide analysis of the spectrin-binding protein in human erythrocytes. One-dimensional and two-dimensional peptide-mapping techniques are used to identify the protein which gives rise to the 72,000 dalton alpha-chymotryptic fragment previously shown to be the membrane attachment site for spectrin. Peptide maps of the 72,000 dalton fragment are very different from maps of Bands 1, 2, 2.9, 3, 3.1, 4.1, and 4.2 and very similar to maps of the apparently closely homologous polypeptides, Bands 2.1, 2.2, 2.3, and 2.6. Limited proteolysis of erythrocyte membranes is shown to generate Band 3', another polypeptide which has been associated with spectrin-binding activity. Peptide maps of Band 3' are very similar to maps of Band 2.1, suggesting that Band 3' is also a proteolytic fragment of Band 2.1. It is concluded that Band 2.1 and possibly some or all of the other, related polypeptides which electrophorese in the 2 region is (are) the spectrin-binding protein(s) of the human erythrocyte.


Peptide mapping of heterogeneous protein samples. A simple two-dimensional electrophoretic method for peptide mapping of heterogeneous protein samples is presented. The reduced and denatured proteins of the mixture are separated in a first dimension by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis. After completion of the electrophoresis, the whole gel lane is equilibrated in stacking gel buffer and is transferred at right angles onto a second slab gel. A protease solution is overlayed on the gel lane and a partial proteolysis of the proteins to be analyzed is performed during the stacking phase of the second electrophoresis. The second electrophoresis resolves the characteristic pattern of peptides of each individual protein as a series of spots located below the original position of the undigested protein. The peptide maps of the following samples are presented as examples: protein P23 and P23* of bacteriophage T4, membranes of Dictyostelium discoideum, membranes of human erythrocytes, and 35S-labeled proteins of D. discoideum synthesized in vivo or in a cell-free wheat germ extract. In complex samples, up to 20 individual proteins can be analyzed at once and a protein comprising only 1% of the total sample generates a clearly identifiable peptide pattern. Good reproducibility of the patterns obtained allows the comparison of samples of different origins.


A novel method of activation of cross-linked agaroses with 1,1'-carbonyldiimidazole which gives a matrix for affinity chromatography devoid of additional charged groups. 1,1'-Carbonyldiimidazole, a carbonylating reagent, has been shown to be suitable for the activation of cross-linked agaroses for affinity chromatography. The activated matrix (an imidazolyl carbamate) is relatively stable to hydrolysis but smoothly reacts with N-nucleophiles such as those present in either affinity chromatography ligands or leashes, e.g. ethylenediamine or 6-aminohexanoic acid. If butylamine was attached via the 1,1'-carbonyldiimidazole method, the resulting product was devoid of charged groups and thus had the same titration curve as agarose. The suitability of this new matrix for affinity chromatography was demonstrated by the successful purification of trypsin by several different systems.


Separation of functionally distinct regions of a macromolecular substrate. Stimulation of tRNA nucleotidyltransferase by a nonreacting fragment of tRNA. Rabbit liver tRNA nucleotidyltransferase catalyzes the incorporation of AMP and CMP into the model acceptor substrate, cytidine. The apparent Km for cytidine in this reaction is about 80 to 90 mM which is more than 10(4) greater than the Km values for the natural substrates, tRNA lacking the terminal AMP (tRNA-C-C) and tRNA lacking the terminal pCpA (tRNA-C). The Vmax values for the model reaction are only 5% and 2% of those for the reaction with the natural tRNA substrates. Addition of the tRNA fragments, tRNA lacking the terminal XpCpCpA sequence (tRNA-(X - 1)p) and tRNA lacking the terminal CpCpA (tRNA-Xp), greatly stimulates the rate of nucleotide incorporation into cytidine. In the case of CMP incorporation into cytidine, tRNA-Xp stimulates the reaction about 60-fold, to a rate similar to that of the normal reaction with tRNA-C. The tRNA fragment has no effect on the apparent Km of either cytidine or CTP, but only alters the Vmax of the reaction. Stimulation of the model reactions is maximal with tRNA fragments of specific chain lengths. These results provide direct evidence that the nonreacting regions of a substrate molecule play an important role in the catalytic efficiency of an enzyme.