The abdominal angiographic spectrum of tuberous sclerosis.

Tuberous sclerosis can present with a variety of clinical, roentgenographic and pathological manifestations. Although we are not advocating abdominal angiography in every patient with tuberous sclerosis, the clinical diagnosis may be difficult in certain cases, particularly in the young patient, and abdominal visceral angiography may lead to the correct diagnosis. In two of our patients, infants ten months of age, the diagnosis of tuberous sclerosis was not made until abdominal angiography had been performed. The most common changes are seen in the kidney where angiographic demonstration of hamartomas, arterial aneurysms, and multiple small cysts in the renal parenchyma are noted. Renal microcysts have not been previously described angiographically although they have been noted in pathological specimens and represent an important feature of the disorder spectrum. In addition, vascular hamartomatous lesions may be demonstrated angiographically in other abdominal organs such as the adrenal gland and liver.


The complication rate of catheter angiography by direct puncture through aorto-femoral bypass grafts.

A search of the literature revealed no documentation of the complication rate associated with percutaneous catheterization of aorto-femoral bypass grafts in the arteriographic evaluation of graft patency. The authors compared the complication rates of 29 consecutive arteriograms performed through grafts (Group I) with 40 arteriograms performed on non patients (Group II). Neither group had any significant complications, indicating the safety of percutaneous catheterization of the graft site.


Systemic arterial supply to normal basilar segments of the left lower lobe.

Two cases of sole arterial supply to normal basilar segments of the left lower lobe via an aberrant systemic vessel are described. A spectrum of congenital pulmonary abnormalities with systemic arterial supply to the lung is presented. Complete roentgenologic evaluation is necessary prior to definitive surgery.


Iatrogenic embolization of the superior mesenteric artery: arteriographic observations and clinical implications.

Following sudden occlusion of the superior mesenteric artery, there is a characteristic and predictable pattern of blood flow through the instantaneously activated collateral channels. On the left, the first patent jejunal or ileal branch utilizes the central tier of the mesenteric arcades to revascularize the distal trunk of the occluded superior mesenteric artery. On the right, the anastomotic connection between the middle colic, right colic, and ileocolic arteries provides an efficient collateral pathway. These two collateral channels supplement each other and the distal trunk of the occluded superior mesenteric artery is usually rapidly reconstituted. Superior mesenteric artery embolization is frequently an asymptomatic event. There appears to be a variable tolerance of cardiac and non-cardiac patients to the effect of superior mesenteric embolization. Non-cardiac patients may tolerate embolization of the superior mesenteric artery quite well. Cardiac patients frequently respond with an ischemic crisis. The site of embolization is another determining factor. An embolus below the origin of the middle colic artery provides an incomparably more favorable hemodynamic situation than an embolus proximal to the origin of this vessel. The excellent tolerance of our group of patients to superior mesenteric embolization appears to be closely related to these two main factors.


Radionclide imaging in skeletal inflammatory and ischemic disease in children.

Sixty-five children were evaluated for presence of skeletal inflammatory and ischemic disease with bone scans and roentgenograms. Several characteristic scintigraphic patterns were observed. Bone scans were significantly more sensitive than roentgenograms in early diagnosis of osteomyelitis and its differentiation from cellulitis, septic arthritis, and bone infarction. The child presenting with possible inflammatory bone disease now is benefited by this important refinement in diagnosis. Faced with the difficult dilemma of choosing appropriate therapy in these frustratingly similar problems, the physician can integrate the clinical findings with nuclear imaging to arrive at early appropriate diagnosis and management.


"Doughnut" sign in brain scanning.

The significance of the "doughnut" sign at present is relegated to characterizing the morphology of a lesion. The use of the "doughnut" sign in differential diagnosis should not be attempted except in specific clinical contexts, and never excluding other evidence. The "doughnut" sign in scanning together with other studies such as angiography and computerized axial tomography gives additional characterizing data in the neuroradiologic evaluation of a patient.


Ultrasonic evaluation of the unilateral nonvisualized kidney.

There are several techniques for evaluating the nonvisualized kidney. Nephrotomography may be helpful in those patients who have some remaining renal function. Radionuclide renal flow and imaging studies are more sensitive than nephrotomography in detecting hydronephrosis, the most common cause of unilateral renal nonvisualization, but also require some renal function to be of diagnostic value. Diagnostic ultrasound, since it is independent of renal function, is an even more sinsitive indicator of urinary obstruction, detecting those cases where no functioning renal parenchyma is present. This non-invasive technique can accurately guide percutaneous puncture of the collecting system, permitting antegrade localization of the obstructing lesion. When ultrasonography demonstrates a solid mass in the renal fossa, angiography is recommended for definitive diagnosis. When no kidney is identified renal venography may be useful in differentiating between a small nonfunctioning kidney and renal agenesis.


Factors that modify the radio-response of cancer of the nasopharynx.

A study of 152 cases of carcinoma of the nasopharynx was carried out for evaluation of various features of the tumors. Survival is greatest in middle aged patients and poorest in patients under 20 years of age. Patients who presented with lymph node involvement had a better survival rate than those without, and those with undifferentiated tumors  survived longer, although tumors in this group metastasized earlier. Those tumors which arose on the postero-superior wall of the nasopharynx were associated with a lower cure rate than those situated elsewhere. Spread to adjacent tissues diminished the cure rate, but the rate fell particularly low if there was spread to bone or to the central nervous system. If no neck lymph nodes were involved, patients with undifferentiated and well-differentiated lesions fared equally well, but if there was involvement of the neck lymph nodes, those with undifferentiated primary lesions obtained a better cure rate. The influence of the type of local spread, the presence of distant metastasis, and survival in relation to the histological and clinical grading of the tumor were also evaluated.


Definitive treatment of carcinoma of the rectum by intracavitary radiation therapy.

A combination of external and contact irradiation treatment has been used successfully in treatment of selected patients with carcinoma of the rectum. The advantages include eliminating a permanent colostomy. If the treatment is unsuccessful in controlling the lesion, electrocoagulation and abdominoperineal resection are still available.


Serial studies of immunocompetence in head and neck cancer patients undergoing radiation therapy.

Studies of the immunocompetence of patients undergoing radiation therapy were carried out before and after treatment. The immunological evaluation is a valuable prognostic tool for patients with squamous cell carcinomas of the head and neck. The degree of immunoresponsiveness is not related to the stage of the primary tumor, but it definitely correlates with the extent of lymph node involvemnt. This evaluation may permit identification of those cases for which immunotherapy is indicated.


Hemangiopericytoma: report of three cases regarding role of radiation therapy.

The experience in treating three patients with hemangiopericytoma is reviewed. One patient was given irradiation alone, and the others were both treated with surgery and irradiation. Our experience reinforces reports of the usefulness of irradiation in treating this neoplasm and its metastases.


Congenital absence of the ductus venosus: with direct connection between the umbilical vein and the distal inferior vena cava.

Umbilical vessel catheterization in a neonate with Noonan's syndrome disclosed a unique developmental anomaly of the umbilical vein. There was no evidence of a ductus venosus nor any connection between the vein and the portal hepatic venous system. The umbilical vein instead entered the inferior vena cava directly at the level of the iliac veins. There was a single umbilical artery.


Aneurysmal bone cyst of the orbit with unusual angiographic features.

A 14 month old boy with sudden onset of proptosis of the left eye due to an aneurysmal bone cyst of the orbit is reported. Sequential plain films of the orbital region demonstrated changes ranging from subtle loss of bone definition of the orbital margin to the characteristic appearance of an aneurysmal bone cyst twelve months later. Arteriography at the time of the initial evaluation demonstrated prolonged retention of the contrast medium which we believe represents changes secondary to the number of giant cells present in the tumor rather than the relative vascularity of the lesion. While aneurysmal bone cyst of the orbit is extremely rare, it should be included in the differential diagnosis of proptosis in the pediatric age group.


Extra-abdominal desmoid of the axillary tail mimicking breast carcinoma.

A case is reported of an extra-abdominal desmoid tumor presenting as an axillary tail breast mass. This represents the third reported case and the first description of the roentgenographic findings of this entity. That this tumor can be confused with carcinoma of the breast is emphasized.


New frontier for radiology: computed tomography. 40th Annual Preston M. Hickey Memorial Lecture.

The basic principles of computed tomography (CT) and their application are discussed. The impact of this method on neuroradiologic diagnosis is reviewed, including economic considerations and the effect on nuclear medicine and other diagnostic procedures. The initial experiences of the Mallinckrodt Institute with whole body scanning are illustrated, and failure possibilities for CT are predicted.


Breast patterns as an index of risk for developing breast cancer.

The radiographic appearance of the breast parenchyma provides a method of predicting who will develop a breast cancer. This paper describes a restrospective study of 7,214 patients. On the basis of the radiographic appearance of the breast parenchyma, patients were placed into one of four groups of risk for developing carcinoma of the breast. Follow-up studies revealed a stepwise progression in the incidence of developing carcinoma of the breast at least 6 months after the radiographic examination. In one of the two substudies, there was a 37 times greater incidence for those at highest risk compared to the low risk group. The classifications presented are thought to be of value in the everyday practice of mammography as well as in planning screening programs.


Radiographic visualization of left ventricular aneurysms on lateral chest film.

The diagnosis of left ventricular aneurysm on routine chest examination can be elusive. Six cases are presented which illustrate a radiographic sign not previously emphasized: a soft tissue border seen retrosternally or superiorly on lateral chest film produced by the wall of the aneurysm silhouetting against the midline cardiac shadow. This sign is not always present since it is dependent on the direction of projection of the aneurysm. However, when observed, it should suggest the proper diagnosis.


The chest x-ray in acute left ventricular power failure: an aid to determining prognosis of patients supported by intraaortic balloon pumping.

Serial chest x-rays were used as a means of evaluating the hemodynamic status of 43 patients in acute left ventricular power failure (LVPF) complicating acute myocardial infarction who were assisted with balloon pumping. The following findings were reported: 1. In patients with acute myocardial infarction, prediction of the hemodynamic status on the basis of chest x-rays is less reliable when severe LVPF is present as a complication. 2. The incidence and severity of roentgenographic findings of congestive heart failure and pulmonary edema are increased in patients with severe LVPF compared to patients with uncomplicated myocardial infarction. 3. Improvement in the roentgenographic degree of heart failure with positive clinical and hemodynamic responses to 24 hr or less of balloon pumping is an indication that patients in severe acute LVPF may survive. Patients with deteriorating or unchanging chest x-ray findings have an extremely poor prognosis. These patients should be evaluated by cardiac catheterization and coronary arteriography to determine the appropriateness of emergency surgical correction.


Abnormal left ventricular catheter motion: an ancillary angiographic sign of left atrial myxoma.

The normal motion of a left ventricular catheter parallels that of the aortic root; it moves anterior during systole and posterior during diastole. In contrast, a prolapsing left atrial myxoma causes paradoxical motion of the catheter; posterior during systole and anterior during diastole. Paradoxical motion was found in each of five cases of prolapsing left atrial myxoma (no false negatives), and in six out of 61 controls (six false positives). In the false positive cases, the catheter was not positioned on the ventricular floor and usually only minor degrees of abnormal motion were present. Paradoxical motion of the left ventricular catheter is an ancillary angiographic finding in prolapsing left atrial tumor.


Evaluation of routine double contrast views of the anterior wall of the stomach.

Since June 1974, we have performed over 1,500 consecutive double contrast upper gastrointestinal examinations. The double contrast study of the anterior wall has been a useful adjunct to conventional mucosal relief and dosed compression studies. Nevertheless, the low diagnostic yield of the anterior wall examination does not appear to justify including it as part of the routine double contrast upper gastrointestinal study.


Intussusception of the appendix in children.

Intussusception of the appendix is uncommon and an unusual cause of ileocolic intussusception. The pediatric age group is most often affected. Appendiceal intussusception may present as primary appendiceal invagination but more commonly leads to secondary intussusception. Two cases of partial appendiceal intussusception leading to secondary intussusception are presented. Classification, symptomatology, radiographic appearance, and differential diagnosis are discussed.


Plain film findings of gastric volvulus herniating into the chest.

When confronted with a chest radiograph demonstrating two large air-fluid levels, one in the retrocardiac mediastinum and the other beneath the left hemidiaphragm, the possibility of gastric volvulus herniating into the thorax should be strongly considered. These patients should have an immediate contrast study for further evaluation.


Chondrosarcoma of the esophagus.

A 46-year-old man had a large noncalcified tumor in the wall of the thoracic esophagus narrowing its lumen by projection of numerous firm nodular masses. The pathologic diagnosis was chondrosarcoma. Malignant degeneration of a tracheobronchial cartilaginous remnant is the main consideration in this first recorded case of esophageal chondrosarcoma.


Localized giant pseudopolyposis in ulcerative colitis.

A case of localized giant pseudopolyposis in ulcertative colitis is discussed in which a mass effect mimicked tumor. The fulminant collection of pseudopolyps was palable in the epigastrium on physical exam and caused a partial obstruction to the retrograde flow of barium. Carcinoma is a worrisome possibility in ulcerative colitis, but localized giant pseudopolyposis may also present as a mass.


Clinical and radiographic "reankylosis" following hip surgery in ankylosing spondylitis.

Eleven patients with ankylosing spondylitis underwent reconstructive hip surgery (21 hips). In 10 of these hips multiple surgical procedures had been performed. The final procedure included total hip arthroplasties (16 hips), femoral cup arthroplasties (four hips) and an Austin-Moore prosthetic replacement (one hip). A clinical and radiographic evaluation in the postoperative period revealed a high incidence of decreased joint motion and heterotopic ossification. Clinically moderate to severe restriction of motion was noted in 12 hips, and in six of these "reankylosis" was present. Radiographically moderate to severe new bone formation was seen in 11 hips, and in nine of these "reankylosis" was suggested. An association of excessive ossification and multiple surgical procedures was evident. It would appear that when the prime indication for hip surgery in patients with ankylosing spondylitis is restricted motion, the operation may not be beneficial.


Medial deviation of the upper pole calyx on the intravenous urogram as an indication of vesicoureteral reflex.

Eighty-three intravenous pyelograms were studied to evaluate thinning of the parenchyma on the medial edge of the superior pole of the kidney as an indicator of vesicoureteral reflux. The data indicate that this finding is suggestive of reflux: the greater the difference between the two sides, the more likely the patient is to reflux.


Ultrasound diagnosis of posttransplant renal lymphocele.

Two cases of posttransplant renal lymphoceles were diagnosed and followed by ultrasound. Lymphocele formation is a significant but often not thought of cause in a rejectionlike picture. Because of its noninvasive nature, ultrasound is an ideal method to detect and follow this complication.


Pseudotumor of the kidney: a sequel to regional glomerulonephritis.

An unusual case of chronic renal failure associated with an abnormal intravenous urogram simulating renal neoplasm is discussed. Extensive radiographic and histologic studies support the contention that the observed mass lesion resulted from a segmental or regional compensatory hypertrophy. It is likely that this case represents a variant of unilateral glomerulonephritis with sparing of a segment or lobe of one kidney and subsequent hypertrophy of that segment as the remaining renal mass atrophied.


Tortuosity of the cavernous carotid arteries causing sellar expansion simulating pituitary adenoma.

Tortuous, medially-displaced cavernous carotid arteries may cause sellar enlargement which simulates pituitary adenoma. Systemic hypertension appears to account for this tortuosity in some cases, while a congenital anomaly is probably responsible in others. Medical position of the carotid sulci may be demonstrated on sellar tomography. Cerebral arteriography provides the correct diagnosis. Surgery, particularly transsphenoidal hypophysectomy, should be avoided in such patients.


Serial cerebral angiography in stereoscopic magnification.

A ceiling x-ray tube support with synchronous up and down movements is described for obtaining two parallel x-ray beams with a short focal-film distance. Stereoscopic cerebral angiography is performed with direct serial magnification using a heavy-duty anode tube with a 0.1 mm focus. This technique combines the advantages of magnification and stereoscopic angiography. Small arterial and venous changes can be observed clearly in three dimensions. The usefulness and limitations of this method are discussed using clinical and experimental data.


Radiologic changes in the thymoma-hypogammaglobulinemia syndrome.

Radiologic findings in the thymoma-hypogammaglobulinemia syndrome are encountered in three stages reflecting the natural history of the syndrome. Initially, a mediastinal mass is apparent on chest x-ray in an asymptomataic patient. At this stage a failing immune system has apparently induced compensatory thymic hyperplasia which progresses to frank neoplasia. In 55% of the cases this thymic tumor is paracardiac in location. With progressive failure of immunoglobulin production, acute and chronic pulmonary infiltrates occur secondary to infection. Markedly, decreased IgA in the bowel may eventually be manifested by radiologic changes of exudative enteropathy.


Cerebro-costo-mandibular syndrome: long term follow-up of a patient and review of the literature.

The cerebro-costo-mandibular syndrome is a rare disorder, and most afflicted infants die during the first few months of life. The pertinent features of the syndrome are reviewed, and a hitherto undescribed finding in the pelvis is described. A case is reported in which vigorous treatment permitted survival. Serial roentgenograms subsequently demonstrated a progressive diminution of the characteristic posterior rib-gap defects.


Intestinal perforations secondary to nasojejunal feeding tubes.

Infant alimentation by nasojejunal tube feeding has been successfully used since 1973. A significant complication is small bowel perforation by the tube; three cases are presented. The differential diagnosis includes spontaneous gastric perforation and necrotizing enterocolitis. Nasojejunal tube perforation almost always occurs in the distal duodenum. It is likely the result of peristaltic activity which propels the tube along the relatively rigid duodenal loop. Thus it is preferable to position the tube end distal to the ligament of Treitz.


Lesion conspicuity, structured noise, and film reader error.

The concept of conspicuity is introduced in an attempt to objectively quantitate radiographic observational error. Defined as a ratio between lesion contrast and surround complexity, the measure correlates well with the probability of detecting faint nodular lesions in chest radiographs. The concept helps in understanding why abnormalities are missed by radiologists. It is also used to explain why image-processing techniques advocated in the past did not yield improvements in diagnostic accuracy and to outline directions for the future. Preliminary results are presented which show that photographic subtraction can increase the conspicuity of simulated early lung lesions and improve their detection.


A circular test pattern for evaluating x-ray tube focal spots.

The design and use of a circular x-ray resolution test pattern is presented. This pattern is shown to yield equivalent focal spot dimensions equal to those obtained with the more commonly used star pattern. The advantage of using the circular pattern is that it obviates the necessity of positioning the pattern strictly along the x-ray beam central axis. A major source of experimental error in making focal spot measurements is therefore circumvented, especially in the case of x-ray tubes with a small target angle.


Lack of gallium uptake in primary hepatic amyloidosis.

99mTc-sulfur colloid and 67Ga citrate hepatic scintigrams showed matching defects in a patient with diffuse primary amyloidosis. Amyloidosis should be added to the usual differential diagnosis of such matching lesions which includes cysts, fibrosis, most benign tumors, and occasional metastatic lesions which do not concentrate gallium.


Incidentally detected thrombosed vein during 67Ga citrate scanning.

A patient with stage IVA Hodgkin's disease developed thrombosis of the right cephalic vein at the site of an indwelling catheter. A few weeks later, 72 and 96 hr scans after injection of 67Ga citrate revealed an increased linear uptake where the thrombosed vein was palpated. As far as we know, this is the first case showing abnormal uptake of this agent in a thrombosed vein. It is postulated that residual chronic inflammatory cells are responsible for 67Ga citrate uptake in this case.


Electron beam therapy in head and neck tumors.

Electron beam therapy with its high energy capabilities appears promising as a therapeutic modality in head and neck neoplasms. Over a 10-year period, 153 cases of head and neck neoplasms involving the alveolar ridge, floor of mouth, buccal mucosa, retromolar trigone, tonsil, parotid, tongue, and pharynx were treated. The pharyngeal group was removed from consideration because of the unsuitability of this lesion for single port therapy. Normal standard doses and time-dose fractions are included as a basis for treatment comparison, and the influence of stage on survival is noted. Survival and tolerance of this form of treatment is comparable to photon therapy and may in fact result in fewer complications.


Surgical treatment of advanced carcinoma of the floor of the mouth.

The anterior arch of the mandible is essential for satisfactory oral function and appearance. It can be preserved in surgical treatment of advanced cancers of the floor of the mouth by partial thickness resection. Surgery and radiation are approximately equally effective in the control of advanced (T3) primary lesions. A treatment plan is proposed which employs a combination of nondisabling surgical resection followed by immediate postoperative radiation. Of 19 patients with T2, T3, and T4 lesions treated and followed 1 year or more, 11 were free of cancer, including six who had positive lymph nodes. Of those dying of cancer, only one had local recurrence, while recurrence in the neck occurred in four. This report further strengthens the need for postsurgical radiation in certain situations.


Central axis depth dose for a 2.5 MV Van de Graaff generator.

Central axis percentage depth dose values and isodose curves for the bremsstrahlung beam from a 2.5 MV Van de Graaff generator were measured with a water phantom at 100 cm target-to-surface distance. Tissue-air ratios were calculated from the central axis depth dose data. Use of the 2.5 MV percentage depth dose values are necessary for treatment planning since they are substantially larger than the values given in compilations for 2.0 MV beams.


Significance of glucose tolerance as prognostic sign in hepatectomized patients.

Glucose tolerance was determined in fourteen patients with cancer of the liver. The patients with parabolic GTT patterns fared relatively better than did those with linear GTT patterns. In the patients with successful extended right hepatectomy, gradually increasing and long-standing hyperglycemia (linear GTT pattern) in response to an oral glucose load was not observed after hepatectomy and the insulin response was significantly greater than that in controls. However, in the patient with unsuccessful extended right hepatectomy, the linear GTT pattern continued for a more prolonged period after hepatectomy.


Hormone effects on human mammary cancer in organ cultures.

Specimens of human breast cancers and fibrosarcomas were incubated for 4 to 72 hours as organ cultures. The effect of various concentrations of testosterone, progesterone, and 17-beta-estradiol on cell survival and multiplication was measured as changes in incorporation of H3-TdR into DNA. Both types of tumor reacted to the hormones; both stimulations and inhibitions were recorded. Statistical analysis of data demonstrated possible influence of factors other than hormone treatment. The specificity of hormone sensitivity assays in vitro is questioned.


Total pancreatectomy for ductal carcinoma of the head of the pancreas: current status.

Because of dismal results after pancreaticoduodenectomy for adenocarcinoma of the head of the pancreas, a review was made of patients treated by total pancreatectomy for this condition. Although the early experience with total pancreatic resection was disappointing, the results during the past decade have been encouraging. Among the forty-two patients reported on with sufficient detail for analysis the operative mortality has been 17 per cent, and the morbidity has been 21 per cent. Sixty-five per cent of these patients (20 of 30) have  survived at least one year and 53 per cent (16 of 30) have lived at least two years after surgery. The management of the diabetes and pancreatic insufficiency has not been difficult. Total pancreatectomy appears to be preferable to pancreaticoduodenectomy in the treatment of ductal carcinoma of the head of the pancreas.


Giant insulinoma.

A patient with an extremely large insulinoma but short duration of symptoms is reported on. Fasting hypoglycemia, hyperinsulinism, and angiography were the modalities used to diagnose and locate the tumor.


Catecholamine-sensitive adenylate cyclase in the developing golden hamster palate.

The present study was initiated to determine whether specific hormones would influence adenylate cyclase activity within the maxillary-palatal complex during formation of the hamster secondary palate. Stages from initial appearance of the palatal processes to shortly after birth were studied. Highest basal adenylate cyclase activities occurred during the earliest periods of palate development. This basal enzyme activity began to diminish as palatal fusion occurred and remained lowered until birth. Activation of adenylate cyclase by fluoride was maximal at concentrations of 5-10 mM, and was observed throughout the span of palatal development. Fluoride activation of adenylate cyclase was greatest prior to fusion of the palatal processes, then decreased until birth when a slightly increased enzymatic stimulation was seen. Norepinephrine and epinphrine were the catecholamines most capable of inducing increased activation of adenylate cyclase at most periods of palatal growth. Increased enzyme activity in the presence of norepinephrine was more susceptible to antagonism by the beta adrenergic agent, propranolol, than to the alpha adrenergic agent, phentolamine. The remaining catecholamines, namely isoproterenol and dopamine, displayed a lesser ability to activate the enzyme, and adenylate cyclase was not equally responsive to these catecholamines at identical developmental stages. Other hormones, i.e. histamine, serotonin, thyrotropin, growth hormone, thyroxine and glucagon were generally ineffective in activating the enzyme. Phosphodiesterase activity was not detected until shortly before birth.


[Cytochrom P-450. Its value in toxicology. I. Physicochemical and biochemical properties].

We present the distribution of cytochrome P450, the terminal oxidase of the microsomal drug-metabolizing system among the different species, organs, and its subcellular localization. Physicochemical properties, and more specially spectral properties, of this hemoprotein are reviewed. We also described the two microsomal electron transport chains and the hypothetical cyclic oxidoreduction mechanism of cytochrome P450 in the hydroxylation reactions.


Immunity to equine herpesvirus type 1 (rhinopneumonitis): in vitro lymphocyte response.

Twenty-two ponies were examined for serum-neutralizing (SN) antibody to equine herpesvirus type 1 and for in vitro lymphocyte transformation in the presence of viral antigen. Six ponies had undetectable levels of neutralizing antibody (titer less than 1:2) and had lymphocytes which did not respond in culture with viral antigen (stimulation index less than 2.0). Four ponies which had SN antibody to equine herpesvirus type 1 did not manifest lymphocyte transformation in vitro. The 12 remaining seropositive ponies had lymphocyte transformation with viral antigen in vitro (stimulation indexes from 2.0 to 23.5). Lymphocyte transformation was specifically suppressed when cells were grown in medium containing autologous serum. The temporal development of in vitro lymphocyte responsiveness and SN antibody in 3 specific-pathogenfree ponies after experimentally induced infection with equine herpesvirus type 1 is described.


Experimental placental transfer of foot-and-mouth disease virus in mice.

An attenuated type O foot-and-mouth disease (FMD) virus which was virulent for infant, but not for pregnant, mice proved to be superior to a virulent type C FMD virus in the development of a model system for the study of placental transfer of FMD in mice. When mice were inoculated at day 8 or 12 of gestation with type O FMD virus, the virus was detectable in the maternal pancreas for 3 days and in the placenta for 6 days. Viral levels in the fetus and the amniotic fluid were inconsistent and were apparently due to a spillover from the placental infection. The elimination of the virus from the placenta coincided with the expected production of maternal 7S antibody. Mice inoculated from days 0 to 12 of gestation did not have a significant increase in dead young by day 18 (the day of necropsy). Similarly inoculated mice, when permitted to go to term, produced and raised normal-size litters. Inoculation on day 15 of gestation resulted in an increased number of deaths due to morbidity of the dams. It was concluded that the placenta serves as an active site of infection for FMD virus in pregnant mice, but the fetus is relatively resistant to infection.


Functional possibilities for aminoacylation of viral RNA in transcription and translation.

RNA from at least ten viruses, representing more than four different group, is known to specifically bind in amino acid in a tRNA-like manner. The biological function of aminoacylation of RNA from these viruses and of tRNA association with tumor viruses and bacteriophage, is discussed. Hypothetical schemes are presented for a role for aminoacylation in viral translational and transcriptional mechanisms, and the current evidence relating to these models is presented.


Interactions of plant viral RNAs and tRNA nucleotidyl transferase.

Ribonucleic acid from TMV and BMV can accept AMP and CMP when digested with VPD. This incorporation is catalyzed by E. coli and yeast tRNA nucleotidyl transferases. Complex formation is obtained between TYMV RNA and tRNA nucleotidyl transferase in sucrose gradients while TMV and BMV RNAs failed to form a complex in the same conditions. The affinity of the enzyme for viral RNAs is lower than the affinity for tRNA as shown by complex formation on nitrocellulose filters and competition with tRNA. Coat protein from TMV particles enhances AMP and CMP incorporation onto tRNA catalyzed by the E. coli tRNA nucleotidyl transferase.


[Reticulohistiocytosis of benign evolution (Hashimoto-Pritzker type). Electron microscopy study].

We describe a rare form of histiocytosis, characterized by numerous tumoral lesions of the skin appearing at birth and desappearing spontaneously within 2 month without visceral lesions and without systemic symptoms. Light microscopy shows histiocytes, some of which contain an irregular nucleus with a spumous cytoplasm surrounded by many red blood cells. Electron microscopy shows myelinoid inclusions in the histiocyte cytoplasm and "vermiform" bodies. In our opinion, a similar anatomoclinical entity was described for the first time and only once by Hashimoto and Pritzker.


[Examination of the peripheral blood in 100 dermatology cases].

Routine examination under the electron microscope of lymphocytes from peripheral blood in 100 patients with skin disease and 20 healthly controls, was carried out seeking branched tubular inclusions of lupus type and lymphocytes with an indented or cribriform nucleus, and gave the following results: -Presence of inclusions of lupus type in 8 cases (4 cases of systemic lupus out of 15 examined, 1 case of sclerodermatomyositis, 3 lymphomas of the skin). These results confirm previous studies carried out in cases of collagen disease and, in addition, demonstrate branched tubular inclusions in lymphomas, they emphasize the importance of the lymphocyte as common denominator in man, in both auto-immune diseases and in lymphoma.-The presence of cells with an indented, or cribriform nucleus in 12 cases out of 100 patients (8 cases of Sézary syndrome out of 8; 4 cases of mycosis fungoides out of 12). Lymphoma deposits were never found outside the skin. -A second type of cytoplasmiclymphocytic inclusion, with a lamellar appearance, has already been described by Hovig and was noted in 13 cases out of 120 biopsies in various diseases and in 2 healthy controls. These were perhaps artifacts.


[Importance of replacing the support medium for viral interferonogenesis in tissue culture].

Interferonogenesis in the cells of chick embryos induced by strain Lee of influenza B virus was studied. The levels of interferon production with and without changing the medium before the cell inoculation with the interferonogens were compared. It was shown that preliminary changing of the medium resulted in a statistically reliable increase in production of interferon as compared to that without the medium changing. It was found that the increase in the synthesis of interferon under the conditions of the maintaining medium changing was accompanied by an increase in the levels of synthesis of the cellular protein and RNA determined by incorporation of C14-leucine and H3-uridine into the acid insoluble fraction of the cells. It is concluded that metabolic changes in the cells at the moment of interferonogenesis induction due to the maintaining medium changing should be considered in estimation of the experiments on the mechanism of the virus induced interferonogenesis in the tissue culture.


[Sensitivity of various races of the tularemia microbe to erythromycin, oleandomycin and polymyxin].

Sensitivity of 66 strains of the tularemia microbe to erythromycin, oleandomycin and polymyxin was studied. It was shown that all the strains were resistant to polymyxin. The strains of the non-artic and Central Asiatic races and 64 per cent of the strains of the holarctic race were resistant to erythromycin and oleandomycin.


[Study of C1. perfringens resistance to lincomycin].

A possibility of developing resistant forms of C1. perfringens during treatment of experimental anaerobic (gaseous) infection with lincomycin was studied. It was shown that treatment of the animals for 7 days resulted in an increase in the resistance by 33-41 times. It was noted that strains with decreased sensitivity to lincomycins had changed morphology and biochemical activity (decreased lecitinase activity, changed biochemical properties), decreased virulence and pathogenicity for animals. So as to obtain the protective effect of the antibiotics in experimental anaerobic (gaseous) infection caused by resistant variants of C1. perfringens it was necessary to increase 1.5-3 times the doses of lincomycin or chlolincocin as compared to the processes induced by sensitive strains.


[Quantitative determination of anhydroleandomycin and glycol in oleandomycin phosphate].

A spectrophotometric method for determination of inactive admixtures in oleandomycin phosphate-anhydrooleandomycin was developed. The method was based on measuring optical density of methanol solutions of oleandomycin and its admixtures at a wave length of 235 nm of UV-spectrum. The minimun detectable amount was 0.5 per cent. A method of thin-layer chromatography in a buffered fixed layer was developed for determination of the 2nd inactive admixture, i.e. glycole. The chromatogrammes were developed with xanthydrol. The minimum detectable amount was 3 mug.


[Effect of migillin on protein synthesis in intact and virally infected TKCE and FL cells].

The effect of migillin was studied with respect to protein synthesis in the cells of TKCE (transplantable line of cells of the kidneys of cow embryos) and FL--intact and infected with viruses of aphtous fever A22, strain 550 and poliomyelitis of type III, strain Saukett respectively. Simultaneously the effect of migillin on sensitivity of the cells to the above viruses was tested. The synthesis of protein was determined by incorporation of C14-glycine into the acid insoluble fraction of the cells. It was found that protein synthesis significantly increased under the effect of migillin in both the intact cells and the cells infected with the viruses. The viruses of aphtous fever and poliomyelitis inhibited the synthesis of proteins. The antibiotic increased the activity of dehydrogenases and respiration of HEp-2 cells. Migillin markedly suppressed reproduction of the poliomyelitis virus, the effect on the virus of aphtous fever was lower in the cell culture. In guinea pigs the antiviral effect of the antibiotic resulted in prolongation of the incubation period and retardation of the aphtous fever process generalization. The results of the experiments showed that migillin stimulated the activity of dehydrogenases, respiration, protein synthesis in the cell cultures and possessed antiviral activity in vitro and in vivo.


Vincristine toxicity unrelated to dose.

Four children with vincristine toxicity unrelated to dose are described. Fever, haematological toxicity, and abdominal distension occurred 2-7 days after vincristine was given. Convulsions occurred 6-8 days after vincristine in all 4. Inappropriate secretion of antidiuretic hormone was thought to have occurred in 3 patients. 2 patients died during the acute toxicity phase. Necropsy findings did not show neuronal changes which could be directly ascribed to vincristine.


Freeze-etching observations of herpes simplex virus.

Herpes simplex virus isolated from the cheek lesions of a patient was propagated on FL cells. Pellets of infected cells were cryoprotected with glycerol, and processed for freeze-etching. Viral nucleocapsids were observed in the nucleus of FL cells. The cores were observed in three forms. Capsomeres were clearly shown. Release of virus particles from the nucleus and the plasmalemma was more clearly observed. Myeloid figures and lipid droplets were also found. The significance of these findings is discussed.


Effects of melanocyte stimulating hormone and theophylline on human melanocytes in vitro.

The effects of MSH on the human melanocyte in vitro were studied. The addition of MSH up to the concentration of 40 mug/ml into the culture medium did not produce appreciable change on the morphology of the melanocyte. The melanocyte, however, responded to the simultaneous addition of MSH and theophylline with marked increase in the length and complexity of the dendritic process. Melanin synthesis, as indicated by the uptake of tyrosine in the presence of an inhibitor of protein synthesis, was remarkably activated by the simultaneous addition of MSH and theophylline. MSH alone activated the melanin synthesis only slightly, but the increase in the uptake of tyrosine was significant statistically. These results were discussed in terms of the MSH-cyclic AMP cascade in which theophylline worked as an inhibitor of cyclic AMP phosphodiesterase, and increased the intracellular level of cyclic AMP by inhibiting catabolism of cyclic AMP.


Functional tumors of the organ of Zuckerkandl.

Chromaffin-reacting pheochromocytomas of the adrenal medulla are the most frequently encountered functional paraganglionic neoplasms. However, extra-adrenal pheochromocytomas as well as non-chromaffin paragangliomas, including those of the carotid body and glomus jugulare, may produce symptoms from catecholamine secretion. One of the extra-adrenal sites from which these tumors arise is from a collection of para-aortic, paraganglion cells around the origin of the inferior mesenteric artery. This collection of paraganglia was described in fetuses by Zuckerkandl in 1901 and has subsequently been referred to as the organ of Zuckerkandl. The diagnosis and management of these neoplasms differ somewhat from that of adrenal pheochromocytomas, but excellent results are often obtained by excision of these lesions. Four patients with functional tumors of the organ of Zuckerkandl are reviewed together with the other reported cases in the literature.


Immunocompetence of patients with bronchogenic carcinoma.

Patients with lung carcinoma were tested for number of circulating thymus-derived (T-)lymphocytes and lymphocyte cytotoxicity before treatment and following operation or radiation therapy or a combination of the two. The proportion of circulating T-lymphocytes in healthy individuals was 75%, and the T-lymphocyte level was 1, 460 per cubic millimeter. All patients with lung cancer had a significantly decreased percentage and absolute number of T-cells before any treatment. Patients tested six to eight weeks following pneumonectomy or lobectomy showed an increased proportion of circulating T-cells in comparison to pretreatment values. Functional activity of immune lymphocytes as shown by cytotoxicity assay also improved in comparison to pretreatment values. The potential value of tests for immunocompetence is discussed.


Sleep patterns and gastric acid secretion in duodenal ucler disease.

Five normal volunteers and five patients with duodenal ulcer (DU) disease were studied for five consecutive nights. All subjects underwent placement of a nasogastric tube, continuous collection of gastric juice, and continuous electroencephalographic monitoring of sleep. Gastric juice was collected in 20-minute samples by remote suction (Gomco). Blood samples were drawn every 20 minutes on the third night via an indwelling venous needle. Results showed no significant correlations between the sleep variables and the gastric acid secretion measures or between the sleep variables and serum gastrin levels. Acid secretion decreased from hour 1 to hour 2 in controls and in patients with inactive DU disease, while it increased in patients with active DU disease. Each subject had at least one night of recording in which continuous gastric secretion was less than 0.1 mEq per 20-minute sample. It appears unlikely that the gastric discomfort of DU patients can be attributed to acid hypersecretion triggered by rapid eye movement sleep.


Oxidative phosphorylation in Micrococcus denitrificans: calculation of the P/O ratio in growing cells.

P/O ratios were measured in membrane particles obtained from cells of Micrococcus denitrificans, while growing on different carbon sources. The membrane particles obtained from cells growing actively on glucose, succinate, ethanol and propanol as the carbon and energy sources catalyzed oxidative phosphorylation and yielded respective P/O ratios of 1.4, 1.2, 0.8, and 0.5 with NADH, and 0.8, 0.6, 0.6, and 0.5 with succinate as the electron donors. Not such a difference in P/O ratio is observed in intact resting cells grown with different carbon sources. It is concluded that the influence of the carbon source is probably directed towards the efficiency of oxidative phosphorylation in membrane particles and not in the growing cells. For the aerobic carbon source-limited chemostat cultures the following maximum growth yields were determined: 40.2 and 34.2 for succinate and oxygen, 41.7 and 36.5 for malate and oxygen, 81.4 and 39.4 for mannitol and oxygen, and 77.8 and 43.4 for gluconate and oxygen respectively. With a mathematical model (de Kwaadsteniet et al., in press) the P/O ratio was valued at 1.4-1.7. YATP at mu=0.2 was valued at 8.7-10.9; YmaxATP at 9.6-13.2 and me at 0.6-4.5 for the most precise experiment (gluconate-limited). The calculation of these growth parameters has been discussed.


Influence of several nucleotides on the competence development of Bacillus subtilis.

The influence of adenosine-3',5'-cyclic monophosphate (cAMP) and other nucleotides on the competence development of Bacillus subtilis was studied. The stimulation of competence which can be achieved by exposing physiologically low-competent cells to supernatants from highly competent cultures can be inhibited with different cAMP doses. When the same cells were suspended in a minimal medium with cAMP, varying degrees of stimulatin- of competence were observed depending on the time of addition of the drug. This effect is not specific for cAMP. It appears to be correlated to an increase of the amount of DNA bound to the competent cells. cAMP activities were antagonized by equimolar doses of adenosine-triphosphate (ATP) and guanosine-triphosphate (GTP).


Energy metabolism of Bdellovibrio bacteriovorus. II. P/O ratio and ATP pool turnover rate.

The P/O ratio of Bdellovibrio bacteriovorus, strain Bd 109 Sa, was evaluated by two different methods based on the determination of energy-rich phosphate bonds and either NADH oxidation or oxygen-uptake. P/O values calculated on the basis of NADH oxidation were up to 6, which has to be regarded as being overestimated. P/O values calculated from energy-rich phosphate bonds and oxygen uptake were around 2. The P/O values determined for Escherichia coli B were similar. The loss of phosphorylation efficiency at one site is discussed. The ATP pool turnover rate of Bdellovibrio was 8/min during endogenous respiration and 24/min during substrate respiration. The corresponding values in Escherichia coli B were 3/min and 38/min.


The effect of glucose and manganese on adenosine-3',5'-monophosphate levels during growth and differentiation of Aspergillus nidulans.

The role of cyclic adenosine monophosphate (cAMP) during growth and development of Aspergillus nidulans was investigated. In normal cultures the highest amount of cAMP, expressed on a dry weight basis, was found after 24 h of growth when still more than 5% glucose was present in the medium. After depletion of the medium even a slight fall in cAMP was noted. Glucose concentrations ranging from 0.5-12% resulted in a slight decrease in the amount of cAMP as measured after 24 h of growth. Cultures with manganese deficiency resulted in a low cAMP level after 24 h of growth. However, the exhaustion of glucose in the absence of manganese was connected with a sharp increase in cAMP. This indicates that manganese shortage was not a direct cause of the low cAMP level after 24 h. The amount of cAMP rose with increasing concentration of manganese in the medium until a maximum at 0.25 muM. It is tempting to speculate that this rise in cAMP in the manganese deficient culture is explained by the absence of glucose, that in the control culture is derived from the breakdown of the reserve material alpha-1,3-glucan. Addition of manganese after glucose exhaustion to a manganese deficient culture induced cleistothecium formation. However, they contained only a few ascospores indicating the importance of alpha-1,3-glucan as a carbon and energy source for ascospore formation. The regulation of the level of cAMP by the transport of glucose into the cell or its intracellular concentration is discussed.


[Histological and electron microscopy studies on the adenocarcinomatosis of laying hens].

Histological examination of 187 tumours from 98 hens (laying hybrids of various lines, age 80-123 weeks) showed that highly-differentiated tubulo-alveolar adenocarcinoma was present in 95% of the birds; in 20% it had a scirrhus structure. Histological and ultrastructural features of these carcinomas and their tendency to form secretory granules pointed to an origin from the protein-secreting part of the oviduct (magnum). In the remaining 5% of birds there were non-specific adenocarcinomas (probably ovarian carcinoma) and mesotheliomas.


Primary Krukenberg tumour in pregnancy.

The literature on primary Krukenberg tumours in pregnancy is reviewed. 6 cases are reported and all could be accepted as complying with the international classification of primary ovarian tumours. An additional case is presented with mostly osteoplastic metastases and necrosis of the marrow.


[C-type-like particles in normal human placentas (author's transl)].

Evidence is presented for the presence of C-type-like particles in the syncytiotrophoblast of normal human placentas. In 10 of 110 mainly immature placentas studied both extracellular and budding particles were found.


Local effects of silica on tumor growth inhibition. A histological study.

The subcutaneous implant of a fibrosarcoma covered with silica, in rats, resulted in a temporary retardation of tumor growth. The histological study demonstrated that the silica provoked, within and at the periphery of the tumor, inflammatory foci which altered the composition of the fundamental substance and which promoted the accumulation of principally macrophages in the interior as well as on the surface of the tumoral mass. Besides the macrophages of the host, a number of tumor cells do also phagocyte silica crystals, leading to destruction of all those cells. The authors are of the opinion that the local modifications provoked by silica, could have interfered in the adaptation of the tumor with its nutrient stroma. Of these modifications, the stroma sclerosis seems to be the most effective in the inhibition of tumor growth. Some correlations were suggested between the here described results and those obtained by other workers who have used other inflammatory agents of different nature.


Dilute ethyl alcohol: effect on the sciatic nerve of the mouse.

The histological changes produced in peripheral nerve by topical ethyl alcohol have been infrequently studied in spite of widespread use of this neurotoxic agent in the management of pain and spasticity. In the present study the sciatic nerves of a series of albino mice were exposed to ethanol in concentrations of from 10% to 50% for from 15 to 60 seconds. An immediate physiochemical reaction took place, resulting in splitting of myelin sheaths and swelling of cellular organelles and cytoplasm. Nerves subsequently underwent Wallerian degeneration. A central core of fibers appeared to be normal. The longer exposure times or higher alcohol concentrations increased the extent to which the peripheral rim of altered tissue extended centrally into the nerve. Within altered tissue, all fiber types and sizes were found to be affected to an equal degree. To study the physical properties of alcohol in solution, absolute alcohol was slowly placed at the bottom of a cylinder filled with artificial or normal human spinal fluid and sequential levels of the solution were sampled 30 and 60 seconds after injection. Though hypobaric, alcohol dispersed rapidly so that specimens taken from the bottom of the cylinder were 65% as concentrated as specimens taken from the surface.


Neuropathy in the upper extremity after open-heart surgery.

Eleven patients with upper-extremity neurological abnormalities underwent open-heart surgery performed through a median sternotomy incision. Seven of the 11 patients were referred in the routine manner to evaluate and treat the neurological problem. The remaining four were part of a consecutively studied group of 11 patients examined prospectively to determine the possible presence of abnormalities. Two of these four patients were asymptomatic. All lesions could be postulated to occur within the brachial plexus, the most common area being the median cord, but lesions were also noted in the posterior and lateral cords and upper trunk. The etiology of the problem appears to be stretching injury of the brachial plexus from retraction of the sternum, which in turn causes retroclavicular displacement of the clavicle. However, it is possible that an ischemic neuropathy could result from intraarterial procedures in some of our patients. The possibility that neurologic deficit may occur in the upper extremity should be considered by physicians who may have the opportunity to evaluate patients who undergo open-heart surgery.


Clinical, virological, and pathological features of herpesvirus SA8 infection in conventional and gnotobiotic infant baboons (Papio cynocephalus).

The pathogenicity of a strain of simian herpesvirus SA8 in one month old conventional and gnotobiotic baboons was investigated. Intratracheal inoculation resulted in a mortality rate of 1/5 in the conventional and 1/4 in the gnotobiotic group. Disease became apparent after 3 days and was characterized by respiratory distress, reduced formula intake, weight loss and fever in both groups. Isolation of herpesvirus from the respiratory tract, lymphoid organs, kidneys, adrenals, and CNS was more frequent by explant culturing than by routine procedures. Although there was a significant difference in total white blood counts (WBC), with higher values in conventional vs. gnotobiotic infants, the absolute number of lymphocytes was not different. The lower number of WBCs apparently was due to fewer polymorphonuclear leukocytes in the gnotobiotic baboons. Infection resulted in a leukopenia 5 days post infection (p.i.) and a leukocytosis 10 days p.i. in both groups. The animals, which succumbed, had acute necrotizing fibrinous pneumonia. Intranuclear inclusion bodies typical for herpesviruses were present. All the surviving infant baboons had subacute interstitial pneumonia, when sacrificed 35 days p.i.


Purification of virions and nucleocapsids of herpes simplex virus by means of metrizamide and sodium metrizoate gradients.

The generation of density gradients for ultracentrifuging by freezing and thawing was applied to formation of linear density gradients with sodium metrizoate and metrizamide. Using these gradient materials simple and rapid methods for purification of virions and nucleocapsids of herpes virus were elaborated. Using metrizamide the recovery of infectivity was 10-30 per cent, and the purification of virions measured as reduction of host protein was 1700 times. Using metrizoate, the recovery of nucleocapsids was 30-60 per cent and the purification from host DNA and protein was 900 and 1700 times, respectively.


Correlation of persistent mouse hepatitis virus (MHV-3) infection with its effect on mouse macrophage cultures.

MHV 3 has three distinct effects in different strains of mice: strain A mice are completely resistant, most strains (including C57BL, DBA/2, BALB/c and NZB strains) die of acute hepatitis whereas in certain strains (eg. C3H and A2G) the virus produces a persistent infection with neurological symptoms. In cultures of peritoneal macrophages from susceptible strains, MHV-3 replicated freely, with giant cell formation. No replication was observed in macrophages from strain A mice. In contrast to this full susceptibility or resistance, macrophage cultures from strains of mice in which persistent infections occur showed an intermediate susceptibility, as judged by the intensity of the cytopathic effect, the presence of viral antigens in the cytoplasm and levels of viral replication. Possible ways in which the intermediate susceptibility of macrophages and persistent infections might be related are discussed.


Persistent parainfluenza type 1 (6/94) infection of brain cells in tissue culture.

A state of persistent infection with parainfluenza type 1 virus (6/94 strain) was established in cultures of human and bovine brain cells. Following primary infection of human brain cells, viral cytopathic effect (CPE) and hemadsorption (HAD) depended on the multiplicity of infection. After persistent infection was established the virus rapidly became cell-associated; no CPE occurred and no viral antigen was detectable by HAD, immunofluorescence (FA), or immunoprecipitation. Infectious virus could be recovered only by fusion or cocultivation. This was in marked contrast with infected bovine brain cells, where, following primary infection, little or no CPE occurred. A productive infection rapidly evolved and persisted without CPE, but with 100 per cent HAD and FA positive cells.


The immunoperoxidase technique for rapid human cytomegalovirus identification.

Sixteen cytomegalovirus (CMV) isolates from both ill and healthy patients were identified by the immunoperoxidase technique (IP). CMV detection was accomplished by direct examination of the primary isolate using either direct (DIP) or indirect immunoperoxidase (IIP) techniques. In thirteen of the isolates, confirmation of identification was achieved by indirect immunofluorescence (IFA) and by demonstration of herpes particles by electron microscopy (EM). Further, in four cases of non-CMV alterations in the tissue culture which might be confused with actual infection, the IP test was negative as were the confirmatory tests. The IIP is preferred over the DIP test since the latter shows a certain amount of background stain of uninfected cells. Tissue culture cells showing focal CMV cytopathic effect contained both nuclear and cytoplasmic inclusions stained by IP technique. Nonspecific staining was associated with cytoplasmic inclusion bodies. The IP technique can detect individual cell CMV infection at 24 hours when EM reveals only unenveloped viral particles. It is sensitive, specific, and allows direct identification of infected cells in the primary isolate in as little as 90 minutes. Furthermore, it can be performed in standard isolation tissue culture tubes, whereas IFA requires the transfer of the infected cells onto slides or the routine use of Leighton tubes.


Detection of type-specific antibody to herpes simplex virus type 1 and 2 in human sera by complement-fixation tests.

Type-specific antigens for herpes simplex virus type 1 and 2 were prepared by rigorous absorption of cell extracts with heterotypic immune sera. Type-specificity was demonstrated by immunodiffusion and complement-fixation tests against immune sera prepared in rabbits. Specific type 1 complement-fixing reactivity was detected in eleven of fifteen sera from Roman Catholic nuns and in two convalescent sera from patients with recurrent herpes labialis; these sera had been previously shown to contain neutralising and complement-fixing antibody to herpes simplex virus. Three of the non-reacting sera contained low or absent levels of type-common complement-fixing reactivity and other contained no type-specific neutralising antibody. With the exception of three "acute" sera, specific type 2 complement-fixing reactivity was detected in every convalescent or interim serum obtained from patients with a virologically-proven history of type 2 herpes virus infection. It is suggested that complement-fixation testing using these absorbed type-specific antigens preparations may provide a convenient and rapid method for the identification of type-specific antibody in human sera.


[Enzymochemical organization of aortic baroreceptors].

The topography of the activity of succinate dehydrogenase, cytochrome oxidase, monoaminoxidase and esterase was studied in the arc of the aorta of the monkey, dog, cat, rabbit, rat. It was established that terminal portions of baroreceptors were accumulators of the activity of enzymes of the succinoxidase system connected with the mitochondria of a nervous component of sensory endings and the activity of cholinesterase localized in the zone of terminal branches of the baroreceptors fibres in the underlying specialized tissue. The activity of monoaminoxidase was found in the cytoplasm of Schwann cells of the baroreceptor fibres. No specific esterase was revealed in the baroceptors of the aorta arc of the animals in question. The enzymic organization of the cat's aorta baroreceptors is different from that of the other animals.


[The morphology of lung cancer and background changes according to bronchoscopic biopsy findings].

Results of a pathohistological investigation of bronchoscopy bioptic specimens in 523 patients, including 422 patients with cancer of the lungs, 7--with benign tumours of the lungs, 83,--with chronic nonspecific afflictions, and 11--with pulmonary tuberculosis, are discussed. A histological diagnosis of bronchogenic cancer was established in 210 out of 422 observations. In central cancer positive results of bronchobiopsy (i. i. revealing of tumour) were obtained in 198 (52.1%) out of 380 cases, in peripheral cancer - in 12 (28.5%) out of 42 cases. Investigations of bronchobiopsy specimens made it possible to establish: epidermoid cancer in 123 (58.6%) cases, microcellular anaplastic cancer in 58 (27.6%) of cases, adenocarcinoma in 15 (7.1%) of cases, macrocellular cancer in 1 (0.5%) case. In 13 observations (6.2%) the histological type of cancer was not identified. The specific gravity of epidermoid cancer was consistently greater than in investigations of operation and section material. Analysis of the background changes in bronchobiopsies in which no tumour tissue was observed showed a substantially lower incidence (P less than 0.01) of atypical metaplasia of the bronchial epithelium in chronic nonspecific diseases than in the bronchogenic cancer.


[A case of cytomegalic disease of the newborn].

A generalized form of cytomegalic disease in newborns is relatively rare. According to the author's data, only one case of cytomegalia was observed during post-mortem investigations of 1724 newborns and young children. This was a full-term child which was born with jaundice, generalized petechial rash and hepatosplenomegaly. Histological investigations revealed cytomegalic cells in the lungs, liver, kidneys and parotid salivary glands; in the brain there was noted calcification of the walls of the lateral ventricles. In the stroma of organs lymphoid infiltration with proliferation of histiocytes was observed.


[Plasticity of the nervous system].

The most recent knowledge about the phenomenon of the nervous system plasticity are revised, as much in morphological as in physiological and molecular levels. The neuron morphological and physiological changes opposite to the experience are studied. The nervous system molecular adaptation to the information it receives as the base of all type of plasticity is also considered.


[20 years later. Or, the long-term outcome of so-called mixed tumors of the lacrimal gland].

A report on two cases of long-term recurrence of mixed tumours of the lacrimal glands. The authors assess the problem of the protracted evolution of these tumours and their possible degeneration, and attempt to distinguish the degeneration which follows multiple recurrences of co-existing mixed carcinomatous tumours. They insist on the necessity in these cases of an initial wide neurosurgical exposure of the orbit, so as to perform the widest excision possible.


Intracranial and extracranial nasopharyngeal angiofibroma. A surgical approach.

Two patients with juvenile nasopharyngeal angiofibromas with known or suspected intracranial extension were treated successfully with a surgical procedure. The procedure involves careful preoperative examination with four-vessel carotid angiography followed by preoperative occlusion of the extracranial major feeding vessels. This is followed by craniotomy, direct control of all intracranial feeding vessels, and then total extirpation of the tumor.


Multipotent lipotropic hormones. In search of a pituitary cell producing multipotent LPH.

The affinity for antiserum to the multipotent lipotropic hormone (beta-LPH) was tested by immunohistochemical staining of all known cell types in normal and certain abnormal mouse, rat, and human pituitaries. Results indicate that beta-LPH has ACTH, MSH, LH and StH(GH) immunologically cross-reacting determinants. Affinities of anti-LPH for TtH and MtH (prolactin) were not detected in normal pituitaries, but thyrotropic tumor cells reacted with anti-LPH. Absorption experiments confirm that the single polypeptide hormone of the pituitary, beta-LPH, is coded for ACTH and MSH activities. The multi-functional hormone, LPH probably is secreted by the adrenotropes. In addition to ACTH and MSH, it probably contains other antigenic and biologic determinants. Some of these may accentuate its lipotropic activities; others may be incidental. These are points calling for further correlated structural, biologic, and immunologic investigations.


Cholangiocarcinoma arising in congenital hepatic fibrosis. A case report.

Congenital hepatic fibrosis (CHF) is a morphologic entity characterized by (1) abnormal fibrous septation, (2) preservation of the basic lobular architecture, and (3) hyperplasia and dilatation of bile ductules. Congenital hepatic fibrosis may present as a cause of portal hypertension in adolescents, in young adults, and, rarely, in older adults. It is this form of the disorder that has been emphasized in the literature. However, CHF may also be seen in association with the infantile polycystic kidney syndrome. In the latter, the hepatic disorder is usually clinically silent. Cholangiocarcinoma is a rare complication of comgenital hepatic fibrosis, with only two previously documented cases reported, to our knowledge. We report a third case and suggest that CHF may represent a dysplastic hamartoma that, with prolonged survival, may eventuate in the development of intrahepatic biliary neoplasia.


Intracytoplasmic lumina in breast carcinoma: a helpful histopathologic feature.

Eelectron microscopy has disclosed the presence of intracytoplasmic lumina within breast cancer cells. These structures can be recognized with the light microscope by their sharp, round outlines and thick walls. Their identification in large numbers may provide additional support for the breast origin of a metastatic tumor. Three illustrative cases in which demonstration of intracytoplasmic lumina was diagnostically helpful are presented.


Melanotic neuroectodermal tumour of infancy: report of a case.

A case of melanotic neuroectodermal tumour occurring in a seven-month-old male infant is reported. The aetiopathogenesis, clinical features, treatment and prognosis of the condition are discussed. Tumours of the jaw in infancy are extremely rare, and pigmented tumours still rarer. One such case of pigmented tumour of the maxilla is the subject of this report.


Sleep monitoring: the second manned Skylab mission.

The first objective measurements of man's ability to obtain adequate sleep during prolonged space flight were made during the three manned Skylab missions. EEG, EOG, and head-motion signals were acquired during sleep by use of an elastic recording cap containing sponge electrodes and an attached miniature preamplifier/accelerometer unit. A control-panel assembly, mounted in the sleep compartment, tested electrodes, preserved analog signals, and automatically analyzed data in real time (providing a telementered indication of sleep stage). One subject was studied during each manned mission and, while there was considerable variation among individuals, several characteristics were common to all three: stage 3 sleep increased during the flight and decreased in the postflight period; stage 4 was consistently decreased postflight, although this stage was variable during the flight; stage REM (rapid eye movement) was elevated, and REM latency decreased in the late postflight period (after day 3 postrecovery); and the number of awakenings during sleep either showed no change or decreased during the flight. In only the 28-d mission (Skylab 2) was there a significant decrease in total sleep time; in that case it was a result of voluntarily reduced rest time and was not due to difficulty in sleeping nor frequent awakening. The subject on the 84-d mission (Skylab 4) experienced more difficulty in the first half of the flight, showing a decreased total sleep time and increased sleep latency, but this resolved itself with time. Sleep latency presented no problem in the other flights. While many of the findings are statistically significant, in no case would they be expected to produce a noticeable decrement of performance capability. These findings suggest that men are able to obtain adequate sleep in regularly scheduled 8-h rest periods during extended space flights. It seems likely, based upon these results, that the problems encountered in earlier space flights did not arise from the zero-g environment per se but possibly were a result of more restricted living and working areas in the pre-Skylab spacecraft.


Metabolic and endocrine studies: the second manned Skylab mission.

This study conducted on the crewmembers of Skylab 3 was designed to evaluate the endocrinological adaption resulting from extend exposure to a space flight environment by identifying changes in hormonal and associated fluid and electrolyte parameters. The three men served as their own controls and were on a constant dietary intake. Complete metabolic collections were performed beginning 21 d before the flight, continuing throughout the flight, for 18 d postflight. Changes in fluid and electrolyte balance have been correlated with weight loss, changes in the excretion of aldosterone, vasopressin, and fluid compartments. Inter-individual variability was demonstrated in most experimental indices measured; however, statistically significant patterns have emerged which include: decreases in body weight and ADH, increases in plasma renin activity, and elevations in urinary catecholamines, aldosterone and cortisol concentrations. Urinary sodium was increased in flight but potassium was only slightly changed. Total body exchangeable K was slightly decreased in all three of the crewmen. Total body water and extracellular fluid were decreased postflight in almost all cases. The measured changes are consistent with the prediction that a relative increase in thoracic blood volume upon transiton to the zero gravity environment is interpretated as a true volume expasion resulting in a net fluid loss. This, in association with other factors, ultimately results in a reduction in intravascular volume leading to an increase in renin and a secondary aldosteronism. Once these compensatory mechanisms are effective in reestablishing positive water balance, the crewemn are considered to be essentially adapted to the space environment. Although the physiological cost of this adaptation must reflect the electrolyte deficit and perhaps other factors, it is assumed that the compensated state is adequate for the demands of the environment; however, this new homeostatic set is not believed to be without physiological cost and could, except with proper precautions, reduce the functional reserve of exposed individuals.


Ultrastructural effects of +Gz stress on swine cardiac muscle.

Miniature swine were subjected to 9 +Gz acceleration for 60-120s. Within 2 h following the +Gz force, the anterior papillary muscle was removed and prepared for scanning and electron microscopy. Ultrastructural changes observed in the cardiac myocytes included cellular redistribution of mitochondria and nuclei. Tears in the contractile fibers, bizarre profiles or nuclei, and peculiar membrane-bounded bodies in the cytoplasm also were observed. Hemorrhagic areas were localized around the Purkinje fibers. The T system and plasma membrane appeared unperturbed. The conclusion was drawn that, following high +Gz levels of acceleration, damage to myocardial ultrastructure ensues.


Heat and simulated high altitude: effects on biochemical indices of stress and performance.

Five healthy, military test volunteers were alternately exposed on separate days to sea level, control conditions (22-23 degrees C, 40-50% RH), a hot wet environment (35 degrees C, 90% RH) or stimulated high altitude (4300 m, 22-23 degrees C, 20-30% RH) while carrying out assigned performance tasks. Acute exposure of 7 h to environmental heat consistently elicited significant increments in plasma levels of cortisol, dopamine-beta-hydroxylase, and uric acid while cholesterol and cyclic AMP levels were unaffected. Alternatively, exposure to altitude for the same duration failed to effect significant alterations in any of the aforementioned biochemical factors. Although both environmental stresses caused similar performance decrements, the responses of biochemical indices of stress were consistently related to the environment itself.


Electron microscopical and enzyme histochemical changes in the rat myocardium during prolonged autolysis.

The effect on rat myocardium of autolysis at 19 degrees C, for up to 20 days, was studied by electron microscopy and enzyme histochemistry. The enzymes studied included monoamine oxidase (MAO), sytochrome oxidase (CytO), non-specific esterase ((Ns.E.), phosphorylase (P-ase), succinate dehydrogenase (SDH) and various NAD- and NADP-linked dehydrogenases. The myocardium lost its histochemical P-ase activity within a few hours of autolysis, whereas the activity of all other enzymes remained quite normal for at least about 4 days, except that of MAO and SDH, which were normal for about 8 and 12 days, respectively. The myocardial cells lost activity of various enzymes in a patchy manner during prolonged autolysis and practically all histochemical enzyme activity disappeared within 20 days. The early period of autolysis was accompanied by rapid ultrastructural changes of myocardial cells. During prolonged autolysis the gross architecture of the myocardium was lost gradually by the 12th to 20th days. Mitochondria were the organelles most resistant to the effects of autolysis, and numerous mitochondria with morphologically solid inner and outer membranes were seen among the totally disintegrated myocardium 20 days after death. The loss of P-ase activity coincided with the loss of glycogen. The loss of MAO, SDH and CytO activities was not closely related to the morphological preservation of mitochondria, but, in accordance with other enzymes, was more closely related to the disintegration of the over-all myocardial structure. The present results showed that the architecture of the myocardium, and especially that of the mitochondria, was surprisingly resistant to the effects of autolysis at room temperature. Also several enzymes of the myocardium other than those examined so far maintained quite stable histochemically demonstrable activity during prolonged autolysis. These observations give support to the possibility of making the diagnosis of myocardial infarction at postmortem more accurate than with the present morphological and histochemical routine methods.


[Needle biopsy in gout and pseudogout (author's transl)].

Radiological and morphological findings in advanced arthritis urica and pyrophosphate arthropathy are well known. In contrast, the early changes of synovial membrane in these disturbances of metabolism pose diagnostic problems. With the assistance of various cytological techniques and polarizing microscopical as well as electron microscopical investigation it was examined to what extent needle biopsies can be helpful in the differential diagnosis of gout and pseudogout. In 8 patients with gout and 11 patients with pseudogout synovial fluid and small tissue specimens could be obtained with the aid of the Parker-Pearson needle. Both fluid and tissue specimens were investigated light and electron microscopically. Cell counts were evaluated in a Rosenthal chamber. The differentiation of the cells in stained smears was done by counting 200-600 cells per case. Crystals were identified by polarizing microscopy in wet preparations of freshly aspirated synovial fluid. Polarizing microscopy of synovial fluid detected intra- as well as extracellular urate and pyrophosphate crystals. The wedge-shaped urate crystals and the larger partly polygonal pyrophosphate crystals showed different polarizing microscopical properties and a negative birefringence. The absolute cell counts in gout were higher than those in pseudogout. The relative cell counts of the different cell types in synovial fluid showed more variation in gout than in pseudogout. Cases with acute gout developed a relative leukocytosis in contrast to a relative lymphocytosis in chronic gout. A relative leukocytosis was constant in all patients with pseudogout. Sclerosed areas with scarce and plump villi as well as sometimes hyperplastic and polymorphous synovial cell layers could be demonstrated histologically in the tissue specimens of the needle biopsies in cases with gout. Urate crystals were less frequent in specimens fixed in formalin. The histological alterations in pseudogout were uniform, 2-4 rows of slightly pleomorphic synovial cells lined the inner surface of the joint capsule, sclerosing alterations were less frequent. Pyrophosphate crystals and calcified particles were seen within the synovial lining cells, the connective tissue and the enodthelial cells of the blood vessels in pseudogout specimens. Intra- as well as extracellular crystals could also be demonstrated with the aid of scanning electron microscopy in sediments of synovial fluid in gout and pseudogout. Transmission electron microscopical investigations of synovial tissue specimens detected proliferated and pleomorphic synovial lining cells in gout in contrast to a more monomorphic appearance of these cells in pseudogout. The crystals were washed out during the preparation techniques for transmission electron microscopy so that needle-like empty spaces resulted within cytoplasm of the phagocytic cells. These clefts were surrounded by phagosomal structures and densified cytoplasmic ground substance; sometimes they were also lined by membranes...


Carbon-13 nuclear-magnetic-resonance spectroscopy of whole cells and of cytochrome C from Neurospora crass grown with (S-Me-13C)methionine.

Neurospora crassa cytochrome C biosynthetically labelled with [S-Me-13C]methionine was prepared and analysed by 13C nuclear-magnetic-resonance spectroscopy. The methyl group of methionine is extensively incorporated into an N-trimethyl-lysine-72 residue arise from S-adenosylmethionine transmethylation, and that the methyl carbons of methionine residues are sufficiently close to the haem centre to experience chemical shifts from the ring currents of the tetrapyrrole pi electrons and broadening due to binding of methionine-80 with the haem, as well as interaction of the S-E113C]methyl groups with the paramagnetic iron centre. Although whole cells of the labelled Neurospora produced a 13C resonance at the expected position for the methionyl methyl group most of the methyl label was diverted into N-tetra-alkyl ammonium compounds. After an active state of growth these labelled N-methyl compounds appear, in the main, to be low-molecular-weight derivatives of choline which, if associated with membrane, are in a sufficiently fluid environment to have short rotational correlation times. During a subsequent dormant growth period these compounds become associated to some extent with relatively more immobile phases as a result of membrane binding or an increase in membrane rigidity.


Effects of sulphate-limited growth in continuous culture on the electron-transport chain and energy conservation in Escherichia coli K12.

Growth of Escherichia coli K12 in a chemostat was limited by sulphate concentrations lower than 300 muM. The synthesis of extracellular polysaccharide and a change in morphology accompanied sulphate-limited growth. Growth yields with respect to the amount of glycerol or oxygen consumed were sixfold and twofold lower respectively under these conditions than when growth was limited by glycerol. Sulphate-limited cells lacked the proton-translocating oxidoreduction segment of the electron-transport chain between NADH and the cytochromes, and particles prepared from these cells lacked the energy-dependent reduction of NAD+ by succinate, DL-alpha-glycerophosphate or D-lactate, suggesting the loss of site-I phosphorylation. Glycerol-limited cells contained cytochrome b556, b562 and o, ubiquinone and low concentrations of menaquinone. Sulphate limitation resulted in the additional synthesis of cytochromes d, a1, b558 and c550; the amount of ubiquinone was decreased and menaquinone was barely detectable. Non-haem iron and acid-labile sulphide concentrations were twofold lower in electron-transport particles prepared from sulphate-limited cells. Recovery of site-I phosphorylation could not be demonstrated after incubating sulphate-limited cells with or without glycerol, in either the absence or presence of added sulphate. The loss of site-I phosphorylation in sulphate-limited cells is discussed with reference to the accompanying alterations in cytochrome composition of such cells. Schemes are proposed for the functional organization of the respiratory chains of E. coli grown under conditions of glycerol or sulphate limitation.


Adenosine 3':5'-cyclic monophosphate-dependent protein kinase in brown fat from newborn rabbits. Changes in the binding of adenosine 3':5'-cyclic monophosphate after preincubation of the tissue with noradrenaline or incubation of the enzyme with adenosine triphosphate.

The equilibrium binding of cyclic AMP to a 150-fold purified preparation of protein kinase, when expressed as the reciprocal of bound against the reciprocal of free cyclic AMP, gave a plot consisting of two straight lines. The values of apparent Kb given by these lines were lowered by preincubating the intact tissue with noradrenaline or incubating the enzyme preparation with Mg2+ plus ATP. This effect was reversed by incubating the preparation (which contained some phosphatase impurities) with Mg2+ alone. None of these procedures affected the maximal binding of cyclic AMP. During incubation of the enzyme with Mg2+ plus ATP, the terminal phosphoryl group was incorporated into protein, over 40% being present in the kinase itself. This phosphate was removed during incubation of the preparation with Mg2+ alone. The validity of expressing cyclic AMP binding as a double-reciprocal plot is discussed, and the experimental plots are compared with those derived theoretically. The results suggest that protein kinase in brown fat is present in two forms, one with an apparent Kb for cyclic AMP or approx. 250 nM (dephosphorylation) and one with an apparent Kb of approx. 14 nM (phosphorylated). Preincubation of the tissue with noradrenaline results in phosphorylation of the kinase and an increase from 15 to 45% in the proportion of the higher-affinity form.


The biosynthesis of alginic acid by Azotobacter vinelandii.

The sequence of reactions by which alginic acid is biosynthesized from sucrose in Azotobacter vinelandii was determined both by feeding radioactive individual enzymes involved. Results indicate that the first polymeric substance formed in the synthesis is polymannuronic acid and that mannuronic acid units are epimerized to guluronic acid at the polymer level. Guluronic acid does not appear to be formed at the monomer level, either free or in combination with GDP.


Growth hormone, cyclic nucleotides and the rapid control of translation in heart muscle.

Perfused rat heart incorporated L-[14C]tyrosine into protein at a constant rate for up to 75 min. A purified bovine growth-hormone preparation (1 mug/ml) stimulated the incorporation to a new constant rate that was more than three times the control rate by 10 min after hormone addition to perfusate. The hormone, however, did not alter the intracellular tracer amino acid pool, and the relationship of this to the aminoacyl-tRNA precursor pool is discussed. It is concluded that the increased incorporation largely reflected a rapid increase in protein synthesis at the ribosomes. Measurements of cyclic nucleotide contents during the perfusion showed that these appeared to vary in a systematic way during the perfusion. This strands in contrast with the constant values given by several other parameters measured in this preparation. Futher, the cyclic nucleotide variation seems to be independent of external effectors. The steady-state performance of the heart correlates more closely the [cyclic AMP]/[cyclic GMP] ratio than with the content of the individual cyclic nucleotides. At 10 min after the addition of growth hormone a slight decrese in cyclic AMP content and a large decrease in cyclic GMP were found, suggesting that the hormone's effect in stimulating protein synthesis may be mediated by a decrease in cyclic nucleotide concentrations or an increase in the [cyclic AMP]/[cyclic |p] ratio. The findings are also consistent with an intracellularly directed role for these nucleotides, and the possibility that the cyclic nucleotide changes are an indirect result of growth-hormone action is discussed.


The dexamethasone receptor in the Novikoff Hepatoma. Characterization and changes in concentration and nuclear uptake during glucocorticoid therapy.

Novikoff hepatoma ascites cells, grown intraperitoneally in rats, are shown to possess a high-affinity, low-capacity dexamethasone-binding protein. The receptor protein has an intracellular localization and concentration, association constant (Ka), glucocorticoid specificity and nuclear uptake in the presence of dexamethasone comparable with those of the G-protein of rat liver. During therapy of the tumour-bearing animal with cortisol, marked cyclic variations were observed in the concentration and nuclear uptake of the putative G-protein in the tumor cells; more transient variations were also observed in the Ka value of the receptor protein.


Yeast phosphoglycerate mutate. Cyanogen bromide cleavage and amino acid sequence of an active-site peptide.

The molecular weight and amino acid composition of phosphoglycerate mutase from yeast were determined. CNBr cleavage produced a large (190-residue) fragment and a small (60-residue) fragment. Tryptic and chymotryptic peptides derived from the large fragment were fractionated by ion-exchange chromatography. Peptides from two histidine-containing regions were isolated and the amino acid sequences were determined. Correlation of these data with X-ray-crystallographic evidence shows that the histidine residue in the sequence Arg-Leu Asn-Glu-Arg-His-Tyr-Gly-Asp-Leu-Glu-Gly-Lys is located at the active site.


Chemistry of the collagen cross-links. Nature of the cross-links in the polymorphic forms of dermal collagen during development.

Both the type I and type III collagens present in embryonic dermis are stabilized by the intermolecular cross-link, hydroxylysino-5-oxonorleucine, derived from hydroxylysine-aldehyde, although the type I collagen possesses a significant proportion of dehydrohydroxylysinonorleucine. However, concurrent with the change in the proportion of the two types of collagen during postnatal development there is a change-over with both type I and III collagens to the labile cross-link, dehydrohydroxylysinonorleucine, derived from lysine aldehyde. The results indicate that the change in the nature of the cross-link with development is determined primarily by the change in the extent of hydroxylation of the lysine residues in the terminal non-helical regions rather than being due to the change in the type of collagen.


Studies by electron-paramagnetic-resonance spectroscopy on the mechanism of action of xanthine dehydrogenase from Veillonella alcalescens.

E.p.r- (electron-paramagnetic-resonance) spectroscopy was used to compare chemical environment and reactivity of molybdenum, flavin and iron-sulphur centres in the enzyme xanthine dehydrogenase from Veillonella alcalescens (Micrococcus lactilyticus) with those of the corresponding centres in milk xanthine oxidase. The dehydrogenase is frequently contaminated with small but variable amounts of a species resistant to oxidation and giving a new molybdenum (V) e.p.r. signal, "Resting I". There is also a "desulpho" form of the enzyme giving a Slow Mo(V) signal, indistinguishable from that of the milk enzyme. Molybdenum of the active enzyme behaves in a manner analogous to that of the milk enzyme, giving a Rapid Mo(V) signal on partial reduction with substrates or dithionite. Detailed comparison shows that molybdenum in each enzyme must have the same ligand atoms arranged in the same manner. As with the milk enzyme, complex-formation between reduced dehydrogenase and purine substrate molecules, presumably interacting at the normal substrate-binding site, modifies the Rapid signal, confirming that such substrates interact near molybdenum. The dehydrogenase-flavin semiquinone signal is identical with that of the oxidase but, in contrast, there is only one iron-sulphur signal. The latter gives an e.p.r. spectrum similar to that of aldehyde oxidase.


Studies by electron-paramagnetic-resonance spectroscopy and stopped-flow spectrophotometry on the mechanism of action of turkey liver xanthine dehydrogenase.

Studies by e.p.r. (electron-paramagnetic-resonance) spectroscopy and by stopped-flow spectrophotometry on turkey liver xanthine dehydrogenase revealed strong similarities to as well as important differences from the Veillonella alcalescens xanthine dehydrogenase and milk xanthine oxidase. The turkey enzyme is contaminated by up to three non-functional forms, giving molybdenum e.p.r. signals designated Resting I, Resting II and Slow. Slow and to a lesser extent Resting I signals are like those from the Veillonella enzyme, whereas Resting II is very like a resting signal described by K. V. Rajagopolan, P. Handler, G. Palmer & H. Beinert (1968) (J. Biol. Chem. 243, 3784-3796) for aldehyde oxidase. Another non-functional form that gives the Inhibited signal is produced on treatment of the enzyme with formaldehyde. Stopped-flow measurements at 450 nm show that, as for the milk enzyme, reduction by xanthine is rate-limiting in enzyme turnover. The active enzyme gives rise to Very Rapid and Rapid molybdenum(V) e.p.r. signals, as well as to an FADH signal. That these signals are almost indistinguishable from those of the milk enzyme, confirms the similarities between the active sites. There are two types of iron-sulphur centres that give signals like those in the milk enzyme, though with slightly different parameters. Quantitative reduction titration of the functional enzyme with xanthine revealed two important differences between the turkey and the milk enzymes. First, the turkey enzyme FADH/FADH2 system has a redox potential sufficiently low that xanthine is incapable of reducing the flavin completely. This finding presumably explains the very low oxidase activity. Secondly, whereas the Fe/S II chromophore in the milk enzyme has a relatively high redox potential, for the turkey enzyme the value of this potential is lower and similar to that of its Fe/S I chromophore.


Some properties of an alcohol dehydrogenase partially purified from baker's yeast grown without added zinc.

Alcohol dehydrogenase was partially purified from yeast (Saccharomyces cerevisiae) grown in the presence of 20 muM-MnSO4 without added Zn2+ and from yeast grown in the presence of 1.8 muM-MnSO4. The enzyme from yeast grown with added Zn2+ has the same properties as the crystalline enzyme from commercial supplies of baker's yeast. The enzyme from yeast grown without added An2+ has quite different properties. It has a mol.wt. in the region of 72000 and an S 20 w of 5.8S. The values can be compared with a mol.wt. of 141000 and an S 20 w of 7.6S for the crystalline enzyme. ADP-ribose, a common impurity in commercial samples of NAD+, is a potent competitive inhibitor of the new enzyme (K1 = 0.5 muM), but is not so for the crystalline enzyme. The observed maximum rate of ethanol oxidation at pH 7.05 and 25 degrees C was decreased 12-fold by the presence of 0.06 mol of inhibitor/mol of NAD+ when using the enzyme from Zn2+-deficient yeast, but with crystalline enzyme the maximum rate was essentially unchanged by this concentration of inhibitor. The kinetic characteristics for the two enzymes with ethanol, butan-1-ol, acetaldehyde and butyraldehyde as substrates are markedly different. These kinetic differences are discussed in relation to the mechanism of catalysis for the enzyme from Zn2+-deficient yeast.


The relationship between biological activity and primary structure of troponin I from white skeletal muscle of the rabbit.

1. A series of defined peptides which span the complete sequence were produced from troponin I isolated from white skeletal muscle of the rabbit. 2. Two peptides, CF1 (residues 64-133) and CN4 (residues 96-117) inhibited the Mg2+-stimulated adenosine triphosphatase of desensitized actomyosin. This inhibition was potentiated by tropomyosin and the Mg2+-stimulated adenosine triphosphatase of desensitized actomyosin. This inhibition, unlike that of troponin I and peptides derived from it, was not potentiated by tropomyosin. 4. The most active inhibitor, peptide CN4, was 45-75% as effective as troponin I when compared on a molar basis. The inhibitory peptide, CN4, and also whole troponin I were shown by affinity chromatography to interact specifically with actin. 5. A strong interaction with troponin C was demonstrated with peptide CF2 (residues 1-47), from the N-terminal region of troponin I. Somewhat weaker interactions were shown with peptides CN5 (residues 1-21) and with the inhibitory peptide CN4. 6. The significance of these interactions for the mechanisms of action of troponin I is discussed.


Study on sex-organ development. Oestrogen-receptor translocation in the developing chick Müllerian duct.

After oestradiol administration in vivo, 87-95% of the initial concentration of oestradiol receptor in the cytoplasm of the embryonic-chick Müllerian-duct cell was translocated into the nucleus. The process of translocation depends on the amount of oestardiol administered in vivo. At 6 h after oestradiol administration in vivo, about 30% replenishment of the initial content of the cytosol receptor was observed in the cytoplasm. The Müllerian-duct nuclei, after exposure to non-radioactive oestradiol, exhibit saturable exchange with [3H]oestradiol in vitro. The exchange of oestradiol is temperature- and time-dependent. The optimal temperature and time for exchange are 37-41 degrees C and 2h respectively. The [3H]oestradiol-receptor complex extracted from the exchanged nuclei is present in 5-6S form, and its isoelectric point is 6.8. The number of nuclear oestradiol-binding sites of the developing Müllerian duct are 1.66, 2.22, 2.63, and 2.50 pmol/mg of DNA respectively for embryos of 10, 12, 15 and 18 days. The dissociation constants of the nuclear oestradiol receptor of the four observed developmental stages range from 3.0 to 3.1 nM.