Internal fixation of fractures: evolution of concepts. The recognized goal of any fracture treatment is the restoration of normal function to the injured part(s). Historically, this was first achieved with non-operative techniques and more recently with operative fixation of fractures. Newer systems utilizing the principles of rigid internal fixation and a sophisticated armamentarium have proved useful in fractures of both bones of the forearm, major intra-articular joint fractures and the multiply injured patient. With these methods "fracture disease" (i.e. muscle atrophy, joint stiffness and bony demineralization) is decreased by early mobilization without external support. The penalties for inadequate surgery are nonrigid fixation and/or infection, emphasizing that the operative treatment of fractures is a technique to be mastered.


Plate osteosynthesis of femoral shaft fractures in adults. A follow-up study. In 67 diaphyseal femoral fractures in adults treated by plating, the average follow-up time was 5 years. Sixty per cent of the patients suffered multiple injuries; 69% of the fractures were comminuted. The goal of treatment was rigid internal fixation. However, bridging of the comminuted area with a plate was sometimes necessary to maintain length as well as to encourage reconstitution and revascularization of fragments. Almost all patients had satisfactory end results. Only 2 cases were rated poor. Patients with rigid interfragmentary fixation had few complications, while patients with obvious instability (14 cases) developed 2/3 of the complications. The infection rate was 3%. Some complications might have been avoided by early cancellous bone grafting. The study confirms the need for primary or early secondary bone grafting to ensure bony union if cortical defects, potentially unstable fixation, or devascularized fragments are present.


Drug-related transient dyskinesias.

Drug-related transient dyskinesias. The ability of some antimalarials, especially chloroquine and amodiaquine, to provoke alarming but transient extrapyramidal movement disorders has been reported. The basis of the reported extrapyramidal symptoms has been the cause of some speculation. This paper reviews case reports and presents some data of the regional uptake of chloroquine in mammalian brain as a factor to consider, along with other factors that could influence the action of chloroquine on the extrapyramidal system. A neuropharmacologic mechanism for the transient extrapyramidal disturbances has been formulated.


Aerosol fenoterol by intermittent positive pressure breathing in asthmatic patients. Ten subjects with mild to moderately severe asthma participated in a study of the bronchodilator activity and incidence of side effects of fenoterol aerosol administered by intermittent positive pressure breathing (IPPB). The doses of fenoterol used in the Bird micronebulizer were 0, 0.25 mg, 0.5 mg, 1.0 mg, and 2.5 mg, and these were administered in a randomized, double-blind fashion. The bronchodilator response, assessed by the area under the FEV1 curve, showed mean (+/- SE) values of 1.44 +/- 0.53 1 hr for 0.25 mg of fenoterol to 1.66 +/- 0.56 1 hr for 2.5 mg of fenoterol (p greater than 0.05), longer (p less than 0.01) than the mean placebo response of 0.06 +/- 0.38 1 hr. A dose-dependent increase in tremor was observed for each of the doses of fenoterol. The 0.25-mg dose of fenoterol solution is an appropriate starting dose for the treatment of moderately severe asthma.


A method for the determination of amitriptyline and its metabolites nortriptyline, 10-hydroxyamitriptyline, and 10-hydroxynortriptyline in human plasma using stable isotope dilution and gas chromatography-chemical ionization mass spectrometry (GC-CIMS). A gas chromatography--mass spectrometry (GC-MS) method has been developed to measure amitriptyline and its metabolites nortriptyline, 10-hydroxyamitriptyline, and 10-hydroxynortriptyline in human plasma. Deuterated analogs of each compound were synthesized as internal standards. Isobutane was used as both gas chromatography (GC) carrier gas and chemical ionization (CI) reagent gas. In order to obtain compounds with satisfactory GC and mass spectrometry (MS) properties, the two alcohol metabolites were dehydrated without loss of label during sample preparation. Selective ion monitoring of the MH+ ions of the protio- and deuterio- compounds gave ion ratios which were converted to plasma concentrations using standard curves. For amitriptyline and nortriptyline, which are assayed using multiple deuterated analogs as internal standards, the curves are straight lines. For 10-hydroxyamitriptyline and 10-hydroxynortriptyline, which are assayed using monodeuterated analogs as internal standards, the curves are nonlinear and are analyzed using an iterative computer procedure. Assay sensitivity is 0.5 ng/ml for amitriptyline, nortriptyline, and 10-hydroxyamitriptyline and 1 ng/ml for 10-hydroxynortriptyline. Assay precision and accuracy in terms of percent error are both less than 5%. Following oral administration of a single 75-mg dose of amitriptyline to two subjects, the mean plasma levels of amitriptyline, nortriptyline, 10-hydroxyamitriptyline, conjugated 10-hydroxyamitriptyline, 10-hydroxynortriptyline, and conjugated 10-hydroxynortriptyline were 36, 8, 10, 66, 16, and 46 ng/ml, respectively, at 2 hr after dosing and 3, 4, 0.5, 1, 6, and 17 ng/ml, respectively, at 72 hr after dosing. Analyses of plasma samples from 12 subjects who had been receiving 50 mg amitriptyline therapy three times a day for an average +/- SD of 32 +/- 5 days gave a mean concentration of 81 +/- 40 ng/ml for amitriptyline, 71 +/- 57 ng/ml for nortriptyline, 12 +/- 5 ng/ml for 10-hydroxyamitriptyline, 91 +/- 30 ng/ml for conjugated 10-hydroxyamitriptyline, 82 +/- 27 ng/ml for 10-hydroxynortriptyline, and 176 +/- 64 ng/ml for conjugated 10-hydroxynortriptyline.


Antihypertensive effects of clonidine.

Antihypertensive effects of clonidine. The antihypertensive effect of clonidine given either two or three times a day with a diuretic was compared in a double-blind crossover study in 18 hypertensive patients. In 11 patients given clonidine three times a day there was better control of blood pressure, although the differences in most instances were of the order of only a few millimeters Hg. There were no differences in the frequency or severity of side effects. Clonidine twice daily with a diuretic appears to be adequate for most patients.


A preventive mastectomy with immediate reconstruction for the high-risk breast cancer patient. A surgical technique for simple mastectomy with immediate reconstruction is described; it is a one-stage procedure. Criteria for performing preventive simple mastectomy are presented. The author invites a 10 year study using the criteria and his operative technique to determine the validity of preventive surgical approach.


Reconstruction of the breast without additional skin or muscle flaps. After mastectomy (usually simple or modified radical) many breasts may be reconstructed without additional tissue. However, in any patient in whom the skin flaps are very thin and/or tight, especially if the flaps have been irradiated, we urge use of the thoracoepigastric flap, latissimus dorsi muscle, or myocutaneous flap to achieve a pleasing result.


Nipple-areola reconstruction: a review of techniques. Various techniques of reconstruction of the nipple-areola complex are reviewed. An analysis of the criteria for an aesthetically pleasing nipple-areola complex is made. Symmetry appears to be the single most important factor in achieving the objective. More than color, texture, or the amount of projection, symmetry appears to provide the best overall result. Those techniques that treat areolae in identical fashion are more likely to produce symmetry. Reducing the normal nipple size not only provides donor tissue but reduces the problem of projection, which otherwise interferes with achievement of symmetry.


Nipple-areola reconstruction following mastectomy: an alternative to the use of labial and contralateral nipple-areolar tissues. The techniques described herein were developed to recreate a nipple-areolar complex that avoids the inherent disadvantages of labial and "nipple-sharing" techniques. We have presented a practical, reliable, one-stage method that utilizes other tissues, and has evolved through experience with 14 cases.


The successful use of "high level" PEEP in near fatal Endrin poisoning. The insecticide Endrin is a highly toxic chlorinated hydrocarbon which can cause severe CNS and respiratory derangement. Death usually follows intoxication within 24 hours. A 19-year-old male developed convulsions and gross pulmonary edema after the ingestion of Endrin. His trachea was intubated and constant positive pressure ventilation with PEEP of up to 28 cm H2O was instituted. The patient survived and recovery appeared to be complete. Severe pulmonary edema is not a common finding in patients who have Endrin poisoning. The use of PEEP above 15 cm H2O is a controversial issue; however, we believe that it contributed significantly to the survival of our patient.


Comparison of serum and urine fibrin split products and urinary beta-glucuronidase in the diagnosis of renal transplant rejection. This study compares the usefulness of serum and urine fibrin split products and the urinary enzyme, beta-glucuronidase, in the diagnosis and management of renal transplant rejection. Fibrin split products, determined by a tanned human red cell agglutination inhibition immunoassay, were measured as a reflection of the secondary fibrinolysis from fibrin deposited in the renal microvasculature as a result of rejection. Urinary beta-glucuronidase, expressed as the ratio of enzyme activity to creatinine concentration, was determined by a colorimetric technique following dialysis of urine to remove endogenous activators and inhibitors. Activity of this lysosomal enzyme is thought to reflect tubular injury. Twenty-nine renal transplant recipients (15 from living donors and 14 from cadaver donors) were evaluated. Both serum and urinary fibrin split products and urinary beta-glucuronidase were markedly elevated in the immediate postoperative period, probably reflecting ischemic trauma. Acute rejection occurring within the first three months was associated with elevations of fibrin split products (particularly urine) and beta-glucuronidase. Elevated values returned to normal following successful treatment with steroids and/or heparin, but remained high in the presence of continued rejection. After the first 48 hours post-transplant, in the absence of rejection, values for fibrin split products were within the normal range. Urinary beta-glucuronidase remained elevated if the transplanted kidney was recovering from acute tubular necrosis. Fibrin split products and urinary beta-glucuronidase were usually normal in chronic rejection.


The burn patient: II. Later care and complications of thermal injury. The care necessary for the extensively burned patient is of an intensity, variety and duration most effectively and economically provided by the multidisciplinary staff of a specialized treatment facility. The organization of burn care in the United States has been well regionalized in recognition of this fact. The burn care given and the research carried out at burn units and burn centers has led to the identification of the pathophysiologic changes, the development of new treatments and the improvement in functional result and survival described in this review.


A capillary technique for cloning Amoeba from single cells. Each amoeba is placed in a drop of approximately 40 microliter of a balanced 'medium + food' solution introduced into the central region of a 7 cm long capillary tube which has first been fused at one end. Such amoebae divide to form clones of 20-30 cells without changing capillaries, though additional food may be required after 7-10 days. Experiments on damage induced by chemicals, cell cycle sensitivities and nuclear/cytoplasmic inheritance, which may involve the cloning of some 2-3,000 or more cells can be carried out with comparative ease. Apart from research, the capillary cloning technique has proved useful in the teaching laboratory. It is adaptable, and can be used for Protozoa other than amoebae, even for requiring sterile conditions.


Early and late results of controlled ventilation in flail chest. From 1967 through 1974, a consecutive series of 35 patients with flail chest were treated with intermittent positive-pressure breathing (IPPB). The controlled ventilation contributed to stabilization of the thoracic cage in a favorable position for healing of the fractures. Surgical stabilization of the chest was not attempted in any of the cases. During treatment with IPPB, one patient died from profuse bleeding due to a generalized coagulation disorder, but the remaining 34 were discharged in a satisfactory respiratory condition. A late follow-up study of the pulmonary function one to eight years after the trauma included x-ray films of the trachea and lungs, kymographic studies of the diaphragm, spirometric and radiospirometric testing, and arterial blood gas levels. Eighteen patients were examined. Spirometric testing revealed astonishingly little impairment of the total pulmonary function. The radiospirometric studies with 133xenon showed a significant reduction of the regional perfusion only in five patients (2 to 4.5 pulmonary segments). Kymographic study of the diaphragm gave no further information and was less selective compared with the other tests of pulmonary function. Owing to the encouraging early and late results in this study, early treatment with IPPB is considered to be the method of choice in flail chest with paradoxic respiratory movements.


Right aortic arch with isolation of left innominate artery. A new entity of right aortic arch with isolation of the left innominate artery is described in a three-day-old boy with complete endocardial cushion defect. Embryologically, this type of anomaly will result if the interruption of the embryonic left aortic arch occurs at two sites, one between the left ductus arteriosus and the descending aorta and the other between the ascending aorta and the left common carotid artery. Absence of pulse in the left arm and on the left side of the neck strongly suggests this anomaly. The aortogram and pulmonary arteriogram may confirm the diagnosis of this anomaly during life.


Rapid definitive diagnosis of Legionnaires' disease. We describe a sporadic case of Legionnaires' disease in which the diagnosis was made by direct immunofluorescence of material obtained by percutaneous aspiration of the involved lung via a needle. Employment of this technique among selected patients with suspected Legionnaires' disease would provide for more rapid diagnosis and more prompt initiation of definitive therapy for some patients.


In vitro sensitivity of Proteus organisms to gentamicin and sisomicin. The antibacterial activity of gentamicin and sisomicin was studied in 148 recent clinical isolates of Proteus obtained from patients hospitalized in Athens. Both gentamicin and sisomicin were found to be active with sisomicin generally being the more active of the two; P. mirabilis strains were less susceptible than the indole-positive strains, but P. mirabilis organisms isolated from the respiratory tract were more sensitive to sisomicin than those isolated from the urine. Susceptibility testing with the two aminoglycoside antibiotics was affected by inoculum size and by the test broth used. Sisomicin sensitivity testing with the disc-agar diffusion method and broth dilution method was reliable for the indole-positive strains of Proteus but did not separate all sensitive from resistant strains of P. mirabilis. An in vitro synergism was demonstrated between sisomicin and the semisynthetic penicillin, ticarcillin.


[A new method of anastomosing severed nerves]. Nerve anastomoses glued with "Fribrinkleber" can be protected from tissue plasminogen-activators both by natural and synthetic inhibitors of fibrinolysis whether administered locally or systemically. The glued nerve-anastomoses do not attain the bond strength of sutured nerves, but show less foreign body reaction. Gluing nerves with Fibrinkleber" combined with inhibition of fibrinolysis would seem to be a good method for reuniting severed nerves. It may be especially useful in nerve transplantation if tension is avoided.


[Early changes of the coagulation system in multiple injuries and their modification with heparin and Trasylol]. In a prospective, randomized study 35 patients with multiple injuries were examined for early changes of blood coagulation. Parameters suggesting a consumption coagulopathy were lower in patients who died subsequently than in survivors. No therapeutic influence of low dose Heparin therapy or of the proteinase inhibiting substance TrasylolR could be demonstrated.


[Various blood-foreign body reactions in the heart-lung machine in heart valve replacement and aortocoronary bypass operations]. Blood-foreign material interaction in the pump oxygenator was studied by means of scanning electron microscopy in 3 patients undergoing heart valve replacement (HVR) and 3 patients undergoing coronary bypass operation (CBO). While polyvinyl tubes show an average deposit of 60 thrombi/mm2 with 20--40 microns in size in CBO, bloodgas interface during HVR causes increasing thrombotic deposits (375 Mcthr/nm2, 25--50 microns in size). Distal to the pump the volume of particles may rise up 250 micrometer in HVR and up to 80 micrometer in CBO. In a 35 micrometer pore arterial-line-filter (Pall-Corporation) platelet clumps up to 1 mm in size are removed from circulating blood in HVR, and up to 300 micrometer thrombi in CBO. According to different platelet deposit proximal to the filter, more and larger thrombi can pass through the filter in HVR. Distal to the pump thromboemboli of 30--60 (maximum 90) micrometer in size can be observed in HVR. Particles of 20--35 (maximum 60) micrometer in size may pass through the filter and enter into the patient's circulation in CBO. As the results suggest, extracorporal circulation is more harmful for patients undergoing HVR than for those undergoing CBO.


[Animal studies on the construction of continent ileostomies and colostomies with the use of BioCarbon implants]. Ring-shaped implants of BioCarbon, a recently developed vitreous carbon polymer, were used for construction of continent ileostomies and colostomies in dogs. The stoma was fashioned by suture fixation of the intestine inside the ring, which was then placed subfascially in the abdominal wall. The exterior of the implant could be hermetically sealed with a screw cap. Implantation in 19 dogs performed so far showed a high rate of incorporation of BioCarbon devices.


[Evaluation of H2 receptor blockade with metiamide in stress ulcers in rats]. Restrain stress in rats inhibits gastric secretion and microcirculation. Secretion is further inhibited by metiamide, but mucosal blood flow is restored to normal. During stress the ulcer incidence is high, lower with prophylactic administration of metiamide and least with the latter and preserved gastroduodenal acid transit. No therapeutic metiamide effect can be detected.


[Metabolic activities in isolated hepatocytes as a vitality parameter in the hypothermically preserved liver]. Vital hepatocytes have been isolated from an adult human liver for the first time. Vitality of the parenchymal cells was examined by trypanblue staining, morphological sppearance and energy-requiring metabolic functions such as gluconeogenesis and glycogen synthesis. Following a cold ischemic time of 6 hours metabolic activity had decreased considerably.


[Effect of time of splenectomy on the increase in survival time following kidney transplantation in rats]. Survival time of Lewis-rats with transplanted DA-kidney grafts following splenectomy at different times after transplantation was evaluated. Splenectomy five days after renal transplantation produced a significant prolongation of survival time (p less than 0.05) whereas splenectomy at the day of transplantation did not extend survival time significantly (p less than 0.1).


[Is permanent survival of kidney allograft receivers in weak histo-incompatible combinations dependent on enhancement?]. To study the immunological status of indefinitely surviving recipients of histocompatible (minor histoincompatible) allografts we transplanted Fi or (Fi x LEW)F1-kidneys to LEW-rats. At the same time bilateral nephrectomy was performed. To examine the cellular immune response we carried out local GvHR, microcytotoxicity assay and allorosette-formation test with recipient cells. We also studied lymphocytotoxins in the serum of recipients. To detect a blocking serum factor we used allorosette-formation inhibition test and microcytotoxicity assay. A blocking serum factor could not be found. In spite of stimulation with donor specific skin graft the cellular immune response of prolonged surviving recipients was inhibited. Our results suggest that prolonged survival of minor histoincompatible renal allograft recipients was caused by suppression of cellular immune response.


Sodium nitroprusside induced hypotension and isovolemic hemodilution in dogs. Hypotension at 70 mm Hg was induced with sodium nitroprusside in 6 hemodiluted and 6 control dogs. In contrast to the control dogs, the reduction in mean arterial pressure was not primarily due to the peripheral resistance reduction, but the result of a hazardous fall in cardiac output.


[Organ tolerance and incomplete bone marrow chimera in dogs]. Cyclophosphamide-induced chimeras develop organ tolerance as do radiation chimeras, as long as hemopoietic cells of donor origin are detectable, independent of the degree of chimerism (n = 7). 4 of 7 dogs with reversion of chimerism rejected their kidney grafts within 11 to 29 days. Three of them, however, retained their kidney grafts permanently indicating that a transient chimerism of a few months duration may be sufficient for induction of tolerance to marrow donor organs in Cy-chimeras. The results suggest that the reversion of chimerism in Cy-chimeras may be due to different mechanisms either immunological rejection or a non-immunological substitution of the grafted marrow by the host's own hemopoiesis.


[Fibrinolytic activity in hyperacute rejected lung allografts]. In hyperacute rejected lung allografts a significantly elevated fibrinolytic activity can be demonstrated which could explain why blood flow is only slightly impaired during this process.


Inhibition of granulocyte chemotaxis by hemoglobin in experimental peritonitis. We have shown in in vivo experiments that hemoglobin interferes with the attraction of polymorphonuclear granulocytes into the peritoneal cavity of rats in response to a bacterial inoculum and thus permits bacterial growth. These findings are proportional to the intraperitoneal concentration of hemoglobin. In in vitro experiments the chemotactic response of human polymorphonuclear granulocytes to zymosan activated serum as well as E. coli bacterial factor is inhibited by hemoglobin. While hemoglobin added in a concentration of 4% to the chemotactic factor causes a significant depression of granulocyte chemotaxis concentrations of only 0.01% are sufficient to cause inhibition of chemotaxis when hemoglobin is added to the cell suspension. The spontaneous migration of the cells is not influenced in either experiment.


[Formation of autologous spongiosa in bone layers as a function of revascularization]. In 24 sheep standardized holes were drilled into the tibia. These defects were filled differently with autologous cancellous bone: a) The defect was filled only with cancellous bone or b) it was filled like in (a) but additionally covered by a Milliporefilter layer; c) the defect was filled like in (a) but the bottom of the defect was covered by a Milliporefilter layer or d) it was filled like in (a) but the cancellous bone was sandwiched by Milliporefilter layers. Bine remodelling was adversely influenced by the Milliporefilters, but the inhibition in (b) was two times bigger than in (c).


[Biomechanical studies on the stabilization effect of cortico-spongiosa chips in imperfect osteosynthesis]. Stability and plate strain of internally fixed femoral shaft fractures at different sites were tested under bending and compression loads. Stability of osteosynthesis with compression plate was not acutely affected by stresses encountered when standing on one leg. When fractures with a bone defect were treated by plate fixation stability was improved significantly when lateral support was preserved or when a medial cortico-spongious bone chip was used.


[Evaluation of digital nerve sensitivity following reconstruction]. 1. In the reconstruction of nerves suture and autogenous grafts are employed in a complementary way. 2. The action potential of sensory nerves does not seem to be an objective parameter to evaluate the results. 3. The Moberg-test detects deficits of the pinch. 4. Two-point-discrimination seems to be the most valid quantitative measurement for sensible reconstruction. The examination is simplified by the two-point-star. 5. The Millesi-test shows good correlation to two-point-discrimination.


[Postoperative blood-sugar regulation in pancreatectomized patients using the artificial "beta cell"]. Seven patients who had undergone total duodenopancreatectomy were connected postoperatively to an artificial 'beta cell', which is a computer-regulated, glucose-controlled-insulin-infusion-system. Despite high-calorie parenteral nutrition, the apparatus enabled successful regulation of the diabetes that followed removal of the pancreas in our patients during the immediate postoperative period (7 days). The artificial beta cell is capable of exogenously regulating the carbohydrate metabolism of pancreatectomized diabetics, even in the extreme conditions immediately following operation.


The organization of the ribosomal RNA genes of Chironomus tentans and some closely related species. Southern gel analysis of total DNA from Chironomus tentans showed that the rRNA genes (rDNA) are homogeneous in structure. After cloning in Escherichia coli plasmid pBR313, the rDNA organisation was further studied by restriction fragment analysis and R-loop mapping. No heterogeneity could be detected by heteroduplex analysis of six different cloned rRNA cistrons. R-loop sizes of 1.69 and 3.63 kilobases (kb) were measured for the 18S and 28S rRNA coding sequences. The two spacers are 0.75 and 1.77 kb long. Southern gel analysis showed also a homogeneous rDNA structure for a Canadian population of C. tentans and C. pallidivittatus. The same technique indicated, however, that the rDNA of two other closely related species of C. thummi and C. melanotus is heterogeneous in structure. A possible correlation between this heterogeneity and the presence of heterochromatin in these species is discussed.


Uptake and clearance of inhalation anesthetics in man. The uptake, distribution, and clearance of inhaled vapors is governed by rules of partial pressure equilibration in a multicompartmental system. Since halogenated anesthetic agents are not soluble in water, biotransformation is their only clearance pathway during anesthesia. When apparent steady state is reached, the rate of overall metabolism can be determined from the pulmonary uptake rate. As a result of metabolism, pulmonary uptake increases but the concentration of inhaled vapor in blood and tissues decreases, and only a fraction of uptake is exhaled following anesthesia. Uptake and pulmonary clearance of five halogenated anesthetic agents were studied in 45 surgical patients. The susceptibility to biotransformation increases in the following order: isoflurane, enflurane, halothane, fluroxene, methoxyflurane.


Clinical pharmacology and pharmacokinetics of nicotinic acid. Evaluation of the biochemical fate of nicotinic acid and the pharmacologic responses it produces illustrate the following general principles. 1. Systemic bioavailability as judged by systemic blood and urine levels of a parent drug may not reflect its bioavailability at the liver, a frequent site of important drug action. 2. Since different pharmacologic effects can show different timedose and route-dose responses, and different degrees of tolerance to repeated doses of the same drug, caution is necessary in evaluating pharmaceutical preparations and making dosage recommendations solely on the basis of drug levels and drug metabolism data. 3. Pharmacologic effects requiring large doses might possibly reflect a need for prolonged exposure rather than high levels. In regard to nicotinic acid itself, the preferred hypocholisterolemic preparation for prolonged treatment may be one which yields little or no discernible blood levels of the drug, in contrast to the vascular effects which require rising nicotinic acid levels, and the fibrinolytic effect which requires parenteral dosage and to which tolerance is acutely developed.


Changes in pancreatic somatostatin content in spontaneously diabetic mice, as determined by radioimmunoassay and immunohistochemical methods. A specific RIA for somatostatin (SRIF) was used to determine the SRIF content of the pancreas and hypothalamus in spontaneously diabetic C57BL/KsJ dbdb and C57BL/6J obob mice. In addition, SRIF- and glucagon-containing cells were examined in the pancreatic islets with an immunohistochemical technique. In dbdb mice, immunoassayable pancreatic SRIF content was increased, as was the number of SRIF- or glucagon-containing cells. In obob mice, immunoassayable pancreatic SRIF content was also increased, but no increase was noted in the number of SRIF- or glucagon-containing cells. The hypothalamic SRIF content of either strain was not different from that of controls. These results regarding pancreatic SRIF content were in accord with some but not all previous reports. These differences may be related to the age of the mice or to the conditions in which they were bred. The pancreatic SRIF increase in both dbdb and obob mice may be attributable to hyperglucagonemia, hyperglycemia, or a decrease in insulin action. Further work is necessary to clearly delineate the mechanism.


Motile events in pancreatic endocrine cells. Monolayer cultures of neonatal rat endocrine pancreatic cells were examined by time lapse cinematography. Motion analysis revealed two major types of motile events. On one hand, particles (0.3 micrometers in diameter) tentatively identified as secretory granules were found to undergo back and forth saltatory movements along oriented pathways. The existence of such pathways was statistically validated by the large prevalence of angles close to either 0 degrees of 180 degrees between successive movements of the same particle. The movements occurred at a mean speed of 0.8 micrometers/sec. Glucose (16.7 mM) increased the frequency of such movements. Vincristine (10 microM) caused a progressive inhibition of saltatory movements, which may depend, therefore, on the integrity of the microtubular apparatus. On the other hand, areas of the cell boundary displayed contractile-like movements, which were stimulated by insulinotropic agents, such as glucose and the ionophore A23187. Cytochalasin B also affected this second type of motile event, which is thought to reflect the activity of actin-like microfilaments. These findings suggest that the microtubular apparatus serves as a guiding cytoskeleton for the oriented translocation of secretory granules, whereas the microfilamentous cell web may control the eventual access of the granules to exocytotic sites.


Secretory granules and progesterone secretion by ovine corpora lutea in vitro. To study the relationship between formation and release of Golgi-derived secretory granules and progesterone secretion, slices of ovine luteal tissue were incubated in the presence of LH and/or calcium ionophore A23187. Increases in progesterone secretion in response to LH and/or ionophore were accompanied by a concomitant release of secretory granules. In contrast, in the presence of colchicine, LH-stimulated progesterone secretion was significantly reduced (P less than 0.01), granule formation appeared to be blocked, and there was little evidence of exocytosis. In addition, unusual pleomorphic membrane-bounded saccules containing an electron-dense material were abundant throughout the centrospheric region of cells treated with colchicine. Because of the close parallelism between formation and release of Golgi-derived secretory granules and progesterone secretion, it appears that progesterone secretion may be coupled to exocytosis of secretory granules. Although the exact content and function of the secretory granules described remains to be elucidated, the data obtained are compatible with the notion that they may contain a progesterone carrier protein.


Effect of in vivo pretreatment of rats with a new protein purified from Bordetella pertussis on in vitro secretion of insulin: role of calcium. Rats were injected once iv with an islet-activating protein (IAP), a new protein purified from the culture medium of Bordetella pertussis. Three days later, their pancreases were studied in vitro for insulin secretory responses. As with pertussis vaccine, pretreatment of rats with IAP was effective in enhancing insulin release from pancreas during perfusion or from islets during incubation in response to secretagogues such as glucose and glibenclamide. The alpha-adrenergic inhibition of insulin secretion induced by epinephrine was also reversed by the pretreatment with IAP. 3-Isobutyl-1-methylxanthine caused insulin release due to accumulation of cAMP. This 3-isobutyl-1-methylxanthine-induced insulin release during perfusion was enhanced in a Ca-containing perfusate, but was conversely reduced in a Ca-free perfusate by the IAP pretreatment. Upon the addition of Ca to the Ca-free perfusate, more insulin was released from pancreases of IAP-treated rats than from those of nontreated rats.


Selective suppression of the primary surge of follicle-stimulating hormone in the rat: further evidence for folliculostatin in porcine follicular fluid. Charcoal-extracted porcine follicular fluid (PFF), when injected at 1200 h on proestrus, blocked the primary FSH surge seen at 1700 and at 1830 h without affecting the LH surge. In contrast, if the injection was withheld until 1330 h, the FSH surge occurred in the normal way at 1700 h but was suppressed at the 1830 h autopsy. The suppression of FSH by 0.5 ml PFF at 1200 h had abated by 0400 h of estrus. The blockade of the primary surge of FSH at 1700 and 1830 h did not prevent ovulation, nor did it prevent the expected rise in serum progesterone or fall of estradiol levels. There appears to be a latency of at least 3.5 h, when PFF is administered ip, before FSH can be suppressed. This latency occurs whether the injection is carried out in intact proestrous rats or in acutely or chronically ovariectomized females.


Effect of neurotensin on insulin, glucagon, and somatostatin release from isolated pancreatic islets. The effects of neurotensin on the release of insulin, glucagon, and somatostatin were investigated in isolated pancreatic islets prepared from 3- to 4-day-old rats and maintained in culture for 48 h before use. Islets were incubated for 20 and 60 min in the presence of 3 or 23 mM glucose with or without neurotensin. In 20-min incubations at 3 mM glucose, neurotensin (10-100 nM) increased the release of insulin, glucagon, and somatostatin by 60%, 90%, and 110%, respectively. These increases were not detected in 60-min incubations. Neurotensin (100 nM) inhibited the release of both insulin (by 60-90%) and somatostatin (by 100%) which was induced by 23 mM glucose in 60-min incubations; this inhibitory effect could be detected with neurotensin at a concentration of 1 nM. Neurotensin also significantly inhibited the elevations in glucagon, insulin, and somatostatin release induced by 20 mM arginine. It is concluded that neurotensin exerts a dual effect on the endocrine pancreas in vitro: 1) at low glucose concentration and over short term (20 min) incubations, the peptide stimulates insulin, glucagon, and somatostatin release; and 2) under stimulated conditions (high glucose or arginine), neurotensin inhibits insulin, glucagon, and somatostatin release.


Changes in hypothalamic LH-RH content and blood levels of LH-RH, gonadotropin and estradiol during the preovulatory stage of rat estrous cycle. In order to investigate the sequence of events concerning gonadotropin surge, serum LH, FSH and estradiol concentrations were measured during the rat estrous cycle as well as hypothalamic and blood levels of LH-RH in the preovulatory stage. Normally cyclic female Wistar rats kept on 12 hr light (from 22.00 hr to 10.00 hr) and 12 hr dark were killed at different times of day during each stage of the cycle. The hypothalamus was quickly dissected out, divided into 3 portions (the anterior, middle and posterior) and extracted in 90% methanol. Blood LH-RH was extracted by affinity chromatography prior to radioimmunoassay. The content of LH-RH in the anterior and middle hypothalamus started to decrease between 1.00 hr-3.00 hr, reached its nadir at 6.00 hr on proestrus and recovered to its previous values on estrus. Almost simultaneously blood LH-RH concentration showed an increase of 18.3 pg/ml-8.8 pg/ml between 1.00 hr-3.00 hr, and then fell to less than 1.0 pg/ml at 6.00 hr. On the other hand, serum estradiol level began to elevate on diestrus II followed by its peak at 6.00 hr on proestrus, while the peaks of serum LH and FSH were observed at 8.00 hr and 10.00 hr, respectively. These studies indicate that the elevation of serum estradiol was followed by the release of LH-RH from the hypothalamus and the LH-RH may be responsible for the preovulatory discharge of gonadotropin.


Topical anesthesia in preendoscopic medication: spray versus lozenges. Patients' acceptance of upper fiberpanendoscopy was significantly (p less than 0.001) better when topical anesthesia with oxybuprocaine was performed with a spray than with a lozenge.


Sodium valproate, serum level and clinical effect in epilepsy: a controlled study. Clinical effects at three different serum levels of sodium valproate (VPA) were compared in a triple-blind, multiple crossover trial comprising 13 epileptic inpatients. Patients were selected regardless of seizure type, and all were in concomitant antiepileptic treatment, which was kept constant throughout the study. A significant relationship between the decrease in number of seizures and increasing VPA serum level was demonstrated. The relationship between VPA dose and serum level was curvilinear. Statistical evaluation of patients by seizure type in relation to clinical effect of VPA was only possible for secondary generalized seizures. Between phenytoin, phenobarbital, and carbamazepine and the different VPA serum levels no interactions could be demonstrated. Recorded side effects were always mild and transient. No obvious correlation between side effects and VPA serum level was established.


Evaluation of feasibility of mutagenic testing of shale oil products and effluents. In an effort to gather preliminary information on the potential genetic hazards of proposed or existing oil shale technologies, we have begun a correlated analytical and genetic analysis of a number of test materials. The work is divided into two phases: one deals with known compounds expected to occur in the environment through shale oil production or use; the other deals with actual samples from existing or experimental processes. A fractionation procedure has been applied to crude product and aqueous product material from an oil shale process. Mutagenicity of the various fractions was assayed by using reversion of histidine-requiring auxotrophs of Salmonella typhimurium (strain TA100, base-substitution mutant; TA98 and TA1537, frameshift mutants). In order to incorporate metabolic activation of these fractions and compounds, we used liver homogenates (S-9) from rats induced with Aroclor 1254 in the standard plate assay. Preliminary results implicate chemical constituents of these fractions (identified or predicted) were tested individually for their mutagenic activity and correlated with the genetic monitoring.


Use of Ames test in evaluation of shale oil fractions. Conditions that affect the sensitivity of the Ames assay of complex hydrocarbon mixtures derived from shale oil were studied. Two fractions, one enriched in polynuclear aromatic compounds (PNA fraction), and a second fraction enriched in aromatic and heterocyclic amines (basic fraction), were selected for most of this work because of their comparatively high mutagenicity (i.e., compared with raw shale oil). The crude shale oil, as well as the basic, PNA, and tar fractions were mutagenic against the Salmonella typhimurium test strains, TA98 and TA100. Mutation was dependent on metabolic activation by microsomal (S9) enzymes. Both test strains responded equally well to the crude product and to the basic fraction; however, strain TA100 was more effective than TA198 in demonstrating the mutagenicity of the PNA fraction. The mutagenicity of the tar fraction could be most easily detected after metabolic activation in a liquid medium, as opposed to S9 activation in the top agar of the standard Ames assay. The mutagenicity of the basic fraction or 2-aminoanthracene was also demonstrated by metabolic activation in a liquid medium. In other set of experiments, the effect of chemical composition on the expression of mutagenicity in the standard Ames assay was estimated. Premutagens requiring metabolic activation were added to the basic and PNA fractions, and the numbers of revertants obtained in the presence of the fractions were compared with mutation induced by the compounds alone. The basic fraction did not interfere with the mutagenicity of 2-aminoanthracene and 7,9 dimethylbenz[c]acridine. Moreover, in certain experiments, the mutagenicity of the complex fraction plus the added compound was higher than expected on the basis of assays performed on these materials separately. Conversely, the PNA fraction prevented or strongly inhibited mutation by several polynuclear aroumatic compounds, and an acridine. However, the PNA fraction did not inhibit mutation induced by 2-aminoanthracene. The effect of the basic fraction on stability of the S9 enzymes in the standard Ames test was also determined.


Characterization of porins from the outer membrane of Salmonella typhimurium. 1. Chemical analysis. The three species of channel-forming outer membrane proteins, porins, have been purified to homogeneity from mutant strains of Salmonella typhimurium which produce single species of porin. Purification was by stepwise solubilization with dodecylsulfate or guanidine thiocyanate, gel filtration, and preparative gel electrophoresis. Amino acid compositions and tryptic peptide maps of the three species of porins showed close resemblance, but at the same time clear differences among them. The number of amino acid residues in the porins purified from the strains SH5551, SH6377 and SH6017 were 361, 354 and 345, and their calculated molecular weights 39800, 39300 and 38000, respectively. Amino-terminal and carboxyl-terminal amino acids in all three species of porins appeared to be alanine and phenylalanine, respectively. Neither half-cystine nor hexosamine was found in these preparation of porins. The isoelectric points of porins from the strains SH5551, SH6377 and SH6017, determined by isoelectric focusing, showed slight differences from each other. These results, and the genetic experiments from another laboratory, suggest that the three species of porins in Salmonella typhimurium are distinct polypeptides, probably coded for by distinct structural genes, which might have been derived from the same ancestral gene.


Characterization of porins from the outer membrane of Salmonella typhimurium. 2. Physical properties of the functional oligomeric aggregates. We have purified to homogeneity, from mutant strains of Salmonella typhimurium, the small oligomers of porin that confer permeability channels to artificial vesicle membranes reconstituted from phospholipids and lipopolysaccharide. The molecular weights of the porin oligomers from the strains SH5551 and SH6017 appeared to be 130000 and 125000, respectively, and those of the monomers were 41000 and 37500, respectively, when determined by sedimentation equilibrium in the presence of dodecylsulfate. It was thus concluded that the functional porin oligomers consisted of three identical subunits. The Stokes' radius of the trimer . dodecylsulfate complex was around 5 nm. The trimer bound less dodecylsulfate than the monomer. The trimer . dodecylsulfate complex retained at room temperature the native conformation of porin, which is rich in beta-structure. When the trimers were dissociated further by various treatments, only the porin monomers were recovered in significant amounts, and the permeability-conferring activity was lost simultaneously. We propose, therefore, that the trimer is the minimal functional unit of porin that is capable of forming permeability channels in the outer membrane of Salmonella typhimurium.


Purification and partial characterization of a factor, a mating hormone produced by mating-type-a cells from Saccharomyces cerevisiae. Cells of Saccharomyces cerevisiae exhibiting the a mating type secrete into the culture medium a mating-type-specific hormone activity (a factor), which specifically causes a transient arrest of DNA replication and cell division in cells of the opposite mating type, alpha. Three compounds exhibiting a factor activity have been found in culture filtrates from a cells. The most active compound has been purified more than 10(5)-fold and appears to be homogeneous on the basis of thin-layer chromatography and thin-layer electrophoresis in different systems. We propose that this compound, which exhibits in alpha cells the biological activities that have been attributed to a factor, represents pure a factor. a factor has been characterized as a very hydrophobic undecapeptide with the following amino acid composition: H2N-Tyr (Asx1, Gly1, Ala1, Val1, Ile2, Phe1, Lys1, Trp1, Pro1). Although in their respective target cells the biological effects of a factor and of alpha factor, the corresponding mating hormone of mating-type-alpha cells, are remarkably similar, the primary structures of both hormones appear to be quite different.


Optical probe responses on sarcoplasmic reticulum: oxacarbocyanines as probes of membrane potential. The relationship between Ca2+ fluxes and the ion diffusion potential was analyzed on sarcoplasmic reticulum membranes using oxacarbocyanine dyes as optical probes for membrane potential. 3.3'-Diethyloxodicarbocyanine responds to ATP-induced Ca2+ uptake by isolated sarcoplasmic reticulum vesicles with a decrease in absorbance at 600 nm. The optical change is reversed during Ca2+ release from sarcoplasmic reticulum induced by KCl or by ADP and inorganic phosphate. The absorbance changes are largely attributable to the binding of accumulated Ca2+ to the membrane. There is no indication that sustained changes in membrane diffusion potential would accompany pump-mediated Ca2+ fluxes. A large change in the absorbance of 3,3'-diethyloxodicarbocyanine was observed on sarcoplasmic reticulum vesicles under the influence of membrane potential generated by valinomycin in the presence of a K+ gradient or by ionophore A23187 in the presence of a Ca2+ gradient. The maximum of the potential-dependent absorbance change is at 575--580 nm. The potentials generated by valinomycin or ionophore A23187 are short-lived due to the high permeability of sarcoplasmic reticulum membranes for cations and anions. There is no correlation between the direction and magnitude of the artifically imposed membrane potential and the rate of Ca2+ uptake or release by isolated sarcoplasmic reticulum vesicles.


Impotence secondary to postpuberal hypogonadism. We studied 4 patients who were under consultation for secondary 'impotence coeundi' and who had had normal puberal development (degree V in the Tanner's scale). 2 of them had children. In the diverse diagnostic attempts, basal deficits of FSH, LH and testosterone were repeatedly confirmed. Stimulation with HCG was positive. Stimulation with Gn-RH did not modify the basal deficient values. Stimulation with clomiphene in 1 case was also negative. The determinations of ACTH, TSH, metopirone test and PBI were normal. In the spermiograms there  was oligoasthenozoospermia. We have not found antecedents that justify the hormonal insufficiency, thus we consider that it is a selective deficit of FSH and LH and of hypothalamic or pituitary postpuberal idiopathic genesis. Sexual potency was quickly regained when the patients were treated with HCG. In every case the dose was adjusted to allow therapeutics.


Post-transplantation ureteral stricture: controlateral pyelocalicostomy. An extensive ureteral stricture developed in a patient 10 years after a renal transplantation. A normal controlateral collecting system has been used for a pyelocalicostomy with excellent postoperative result.


Demonstration of HLA-DR-like antigens on milk fat globule membranes. Milk fat globules (MFG), which are formed by exocytosis of lipid from epithelial cells of the mammary gland, are enveloped by plasma membrane from the epithelial cells. Highly purified, detergent-solubilized MFG membranes have been shown to contain molecules reactive with a rabbit antiserum against HLA-DR antigens. Indirect immunoprecipitation combined with polyacrylamide gel electrophoresis in sodium dodecyl sulfate demonstrated that the MFG membrane material reactive with the antiserum comprised molecules which under denaturing conditions displayed molecular weights of 28,000 and 35,000. The two types of polypeptide chains, which were both glycosylated, were held together by noncovalent forces under nondenaturing conditions. Various types of chemical and physicochemical analyses failed to reveal any significant differences between the HLA-DR-like antigens from MFG and from spleen cells. Since the HLA-DR-like antigens bound detergent in micellar form and were expressed on the outside of intact MFG, as revealed by indirect immunofluorescence, it is concluded that these antigens are embedded in the hydrocarbon matrix of the MFG and not merely passively adsorbed onto the MFG.


A human thymocyte antigen defined by a hybrid myeloma monoclonal antibody. Spleen cells from a BALB/c mouse that had been immunized with human thymocytes were fused with the myeloma line P3-NS 1/1 Ag 4.1. One of the resulting hybrid clones (NA 1/34) secreted an antibody that was highly specific for human thymocytes. Eighty-five % of thymocytes expressed the antigen designated HTA1. There were an estimated 15 x 10(4) molecules of HTA 1 per cell, and it is therefore a major surface molecule. The expression of this antigen on thymocytes appears to be reciprocal to HLA, as recognized by another monoclonal antibody W6/32. Immunoprecipitated material from [125I]-labeled thymocyte membranes was analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate which disclosed a single component of 45,000 molecular weight.


Endotoxin-induced myeloid reactions in dogs. The response to artificial endotoxinemia was studied in adult dogs. Granulocyte-macrophage colony stimulating activity (CSA) in lung tissue and blood was measured along with the number of circulating granulocytes and myeloid committed stem cells (colony forming units, CFUc). Acute endotoxinemia induced a measurable CSA increase in lung tissue before being detectable in blood. Granulocytes, rapidly removed from the circulation, showed no release of CSA during sequestration. These experiments demonstrate that the process of endotoxin recognition and subsequent transition into a myelopoietic stimulus is medicated by cells belonging to tissue; mature granulocytes, involved in the defence against bacterial infection, do not release activity that promotes growth of immature myeloid cells.


Proteolytic enzymes in the Rhodnius prolixus midgut. Sequential chromatographic fractionation of Rhodnius prolixus midgut homogenate yielded only one endopeptidase, but revealed the presence of carboxypeptidase-A and B-like enzyme, di- and tripeptidases, as well as aminopeptidase activities.


The effect of polysaccharide-protein complex isolated from Candida albicans on regional blood flow in rats. After i.v. administration to rats of polysaccharide-protein complex, isolated from Candida albicans, a decrease of cardiac output was observed from 20 sec to 240 min postinjection, followed by a recovery at 360 min. Concomitantly the regional blood flow was maintained in heart and lungs, moderately decreased in intestine, liver and adrenals and markedly reduced in skin, muscle, spleen and kidney.


HBsAg uptake by macrophages in vitro: an immunofluorescence study. The uptake of HBsAg by in vitro cultured macrophages was studied by immunofluorescence method. Intracytoplasmic fluorescent particles appeared 3 h after the contact with HBsAg-positive serum, while after 24-48 h only a few cells contained these particles, which are probably destroyed within the cytoplasm.


Asialoprotein uptake by liver cells: immunofluorescence microscopy. Uptake of asialoproteins by hepatocytes causes a change in the intracellular pattern of immunofluorescence. Control cells display a peripheral fluorescence which probably represents nascent proteins. Dark nonfluorescent areas, that presumably contain glycogen, are located around the nucleus. In contrast, liver cells from rats injected with asialoproteins display a pancytoplasmic fluorescence due to an influx of endocytotic vesicles.


Tick-borne encephalitis virus-specified sequences in persistently infected cell culture revealed by DNA-DNA hybridization. Hybridization of DNA probe, obtained through DNA polymerase-mediated in vitro transcription of tick-borne encephalitis virus (TBEV) RNA, with DNA isolated from persistently infected with TBEV cell culture revealed 5.4 copies of viral genome per haploid set.


Myosin-like protein in the human parathyroid glands: an immunohistochemical study. The immunofluorescent staining of the human parathyroid glands shows that the principal cells bind antimyosin-like antibodies. The presence of a myosin-like protein in the principal cells of the human parathyroid glands is presumably in rapport with parathormone secretion.


Evidence for changes in cell shape from a 2-dimensional to a 3-dimensional substrate. Chick embryo mesoderm cells were explanted to culture systems in vivo and in vitro and their subsequent movements were correlated with the external morphology as studied by SEM. In vitro cell movements are exaggerations of normal in vivo movements where a 2-dimensional substrate is encountered rather than a 3-dimensional environment.


[Comparative study of the effect of thymosin and supernatant from epithelial thymic cells on antibody production (author's transl)]. A thymic extract (TE) was prepared from supernatant of mice thymic epithelial cultures according to the purification of thymosin. TE and thymosin stimulated, in vitro, the immune response of mouse against deep red blood cells.


Immunological recovery of thymectomized and sham-thymectomized lethally irradiated mice reconstituted with syngeneic bone marrow cells. Immunological functions of lethally irradiated mice reconstituted with syngeneic bone marrow cells recover after 5--6 weeks. In mice that had been thymectomized before irradiation and reconstitution, T-cell function is deficient but the B-cell function is preserved.


Protective effects of supplemental vitamin E against infection. Vitamin E supplementation (dl-alpha-tocopheryl acetate except where noted) in excess of requirement significantly increased humoral immune response or disease resistance. Mice immunized with sheep red blood cells or tetanus toxoid and fed the supplemental vitamin demonstrated increased plaque-forming cells (PFC) and hemagglutinin (HA) titers. A vitamin E deficiency resulted in decreased PFC and little IgG which was partially corrected by N,N-diphenyl-p-phenylenediamine but not as effectively as by vitamin E. Hens immunized with Brucella abortus and fed different levels of the vitamin produced chicks with increased passive immunity; a biphasic antibody response to the level of the vitamin fed was noted. Vitamin E fed to nonimmunized hens was found to significantly increase the primary immune response of their immunized chicks. Feeding dl-alpha-tocopheryl acetate to guinea pigs immunized with Venezuelan equine encephalomyelitis virus resulted in no increased immunity. Injecting this form of the vitamin resulted in severe tissue reaction. However, injecting dl-alpha-tocopheryl significantly improved hemagglutinin inhibition titers. Chicks and turkeys infected with Escherichia coli and fed supplemental vitamin E had reduced mortality and increased HA titers. Sheep fed vitamin E and challenged with Chlamydia had improved weight gains and no detectable Chlamydia.


The "treatment" of unexplained infertility with danazol. In order to investigate the possible stimulating effect of danazol on fertility, a randomized clinical trial was performed on 40 women with unexplained infertility. Of these 40 women, 21 received 200 mg of danazol daily for 100 days and 19 received a placebo treatment during the same period. Serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin, estrone, estradiol, progesterone, and testosterone were followed before, during, and after treatment. Danazol administration induced anovulation in all women, with prompt resumption of normal ovulatory function after discontinuation of the drug. No influence was seen on serum LH, FSH, and testosterone levels, but serum estrone, estradiol, and progesterone levels decreased significantly during treatment. Serum prolactin levels also decreased, but not significantly. No pregnancies occurred in the placebo group during a 6-month follow-up period. In the danazol group, five pregnancies occurred, of which two were ectopic and three went to term. The difference in pregnancy rate between both groups was statistically significant (P less than 0.05).


Gonadotropin-releasing hormone infusion test in the distinction of hypopituitary patients from normal subjects. We undertook a pilot study to determine whether infusion of gonadotropin-releasing hormone (GnRH) might improve the distinction of hypogonadotropinism from the normal state and might permit gonadotropin deficiency to be diagnosed in the prepubertal child. Normal prepubertal and pubertal boys had a greater luteinizing hormone (LH) reaction (delta LH 54 +/- 15 [SD] ng/ml and 165 +/- 23 ng/ml, respectively) to a 4-hour infusion (100 microgram/hour) than to a 100-microgram bolus of GnRH (19 +/- 9 and 52 +/- 35 ng/ml). These augmented responses were observed in boys with delayed puberty, but not in apparently hypogonadotropic males greater than or equal to 12 years old. LH (delta LH 445 to 1602 ng/ml) and FSH (delta FSH 718 to 2112 ng/ml) surges were induced consistently by GnRH infusion only in normal, postmenarchial females. In all, of 13 hypopituitary cases classified as hypogonadotropic on the basis of a subnormal response to GnRH infusion, 31% had a normal response to the GnRH bolus (P = 0.05). Thus, GnRH infusion testing seems to improve the distinction of hypogonadotropic patients from normal individuals, including boys with delayed puberty.


Chlamydial serology in infertile women by immunofluorescence. The chlamydial serum immunoglobulin G antibodies of 128 infertile women were determined by a single-antigen (L2) immunofluorescence test. The antibody titers were compared with those of female contacts of men with nonspecific urethritis and with those of "normal" pregnant women. Among infertile women, 43% lacked chlamydial antibodies (titer less than or equal to 8). The corresponding proportion for contacts of men with nonspecific urethritis was 14% and that for pregnant women 71%. A very high titer (greater than or equal to 256) was recorded in 25 infertile women and in 7% of pregnant women. Among women with positive chlamydial isolation, 46% had titers greater than or equal to 256. Bilateral tubal obstruction was demonstrated in 23 infertile women (18%). The geometric mean titer of these patients was higher than that of patients with normal hysterosalpingograms. In nine cases with bilateral tubal obstruction and very high chlamydial antibody titers, there was no history of pelvis infections. The operative findings were typical of previous salpingo-oophoritis.


Paradoxical effects of D-Trp6-luteinizing hormone-releasing hormone on the hypothalamic-pituitary-gonadal axis in immature female rats. The effect of administration of a superactive and long-acting analog of luteinizing hormone-releasing hormone (LH-RH), D-Trp6-LH-RH, in doses of 0.05 or 1 microgram/day for 10 days on the hypothalamic-pituitary-gonadal axis was studied in immature female rats. Treatment with a 0.05-microgram dose of analog produced few changes as compared with the control group. Treatment with 1 microgram of D-Trp6-LH-RH did not affect the body weight or the pituitary weight, but increased ovarian weight and decreased uterine weight; elevated serum gonadotropin levels; and lowered the pituitary LH content. This depletion of pituitary LH content was associated with a low pituitary responsiveness to LH-RH. Serum estradiol levels were not modified, suggesting that decreased uterine weight reflects a direct and extrapituitary effect of this analog. The hypothalamic LH-RH content was higher, indicating a possible inhibition of the release of endogenous LH-RH. A delay in vaginal opening was also observed. This indicates that large doses of D-Trp6-LH-RH may interfere with the process of puberty in rats. These findings extend other reports about the paradoxical antifertility effects of large doses of stimulatory analogs of LH-RH.


Glucagon and diabetes mellitus.

Glucagon and diabetes mellitus. The present status of knowledge about glucagon pathophysiology in diabetes is reviewed. 1) A-cells behave abnormally in all varieties of diabetes mellitus, spontaneous and experimental, except perhaps in case of pancreatectomized humans. These abnormalities are : hyperreactivity of A-cells to arginine, non suppressibility by glucose, and absence of stimulation following hypoglycemia. 2) These abnormalities appear as secondary in most instances : a) A-cells behave in a normal way in most studies with prediabetics ; b) plasma glucagon concentration is normalized by excellent control of diabetes or following prolonged insulin infusion. High doses of insulin are required most of the times to obtain a normalization of A-cell function : in insulin-dependent diabetics, the physiological portoperipheral insulin gradient no longer exists, and the high doses of insulin which are necessary may be the only mean to reconstitute the high insulin concentrations supposed to be present at the A-cell level. 3) Conflicting results have been collected about the role of this glucagon excess in aggravating the diabetic metabolic syndrome. Evanescent effects follow sustained glucagon infusions: but in diabetics, glucagon bursts rather than permanent hyperglucagonemia are observed and these appear deleterious to glucose tolerance. It seems clear however that insulin deprivation is required for the full expression of the consequences of glucagon excess.


Anti-idiotypic antibodies induce formation of anti-H-2 sera and of idiotypes. Alloantisera could be induced in parental strain recipients either by means of alloantigen (skin allografting) or by injecting anti-idiotypic (anti-T cell receptor) sera. The resulting sera displayed activity against H-2 antigens; they also contained soluble isiotypic structures. With these sera anti-idiotypic sera could be provoked in F1 hybrid hosts. If antibody cycles initiated with alloantigen were carried through four runs, specificity appeared to be preserved and titers obtained corresponded to those found in conventionally raised anti-idiotypic sera and in alloantisera.


Inhibin--fact or artifact.

Inhibin--fact or artifact. Peripheral concentrations of follicle-stimulating hormone (FSH) in male as well as in female animals appear to be partly regulated by inhibin, a protein which is secreted by the gonads. The molecular structure of this substance is still unknown, and the mechanism(s) of its action on the pituitary or hypothalamic level is not clear. Much of the confusion about inhibin stems from the fact that no generally accepted definition of inhibin exists and that fundamentally different biological assay systems have been used by different groups. Therefore this short review starts with a discussion of the definition of inhibin and the assay principles. From the available information on the site of origin of the hormone it appears likely that inhibin is produced in the Sertoli cells of the testis and the granulosa cells of the ovary. The available data on the chemical nature of inhibin suggest that different principles, acting on different sites of the hypothalamic--pituitary axis, might be present in preparations with inhibin-like activity. Finally, with respect to the biological significance of inhibin, it seems that inhibin could play a more important role in the feedback regulation of FSH in the adult female than in the adult male animal.


Effect of fasting on the release of insulin and somatostatin from perifused islets of Langerhans. Release of somatostatin and insulin from perifused islets of fasted and control rats was compared. After a fasting period of 48 h glucose-induced insulin release but not somatostatin release was diminished. Islets from fasted rats released significantly more somatostatin in the presence of 3.3 mM glucose than islets from controls. Simultaneously, the somatostatin content of isolated islets from fasting rats was significantly decreased. The results indicate that the low secretory activity of islet B cells in the fasting state is associated with a high secretory activity of islet D cells.


Correlation between morphology and function in isolated islets of the Zucker rat. Obesity in the Zucker rat is accompanied by hyperlipemia, hyperinsulinism, insulin resistance, pancreatic hyperplasia, and islet hypertrophy. This study correlates the morphologic heterogeneity of isolated pancreatic islets with secretion of insulin and glucagon in the perifusion system. Islet size was arbitrarily defined as large (greater than 0.45 mm) or small (smaller than 0.12 mm). Protein content and volume (V = 4/3pir3) were calculated for groups and individual islets, respectively. Islets from obese rats secreted more insulin in response to glucose and aminophylline than islets from lean rats (peak 7.8 +/- 2.4 vs. 1.5 +/- 0.37 microU/islet/min, P less than 0.005). Insulin release was related directly to islet size and protein content. Small islets from lean and obese animals produced less insulin per islet than large islets (P less than 0.005). In terms of islet volume, however, large islets were inefficient insulin releasers as compared to small islets (P less than 0.005). Stimulation with Br-cAMP released glucagon from islets of lean but not from large islets of obese animals (peak 11 +/- 3.3 vs. 4.1 +/- 0.3 pg/microgram protein per minute, P less than 0.05). Arginine produced the same effect on glucagon release (P less than 0.05) as stimulation with Br-cAMP. The observed increased insulin release rates and the blunted glucagon response are related to islet size in the pancreas of the Zucker rat.


Effect of OK-432 on immunization with mitomycin-C-treated L1210 cells. The pretreatment of B6D2F1 mice with OK-432 alone was not so effective to retard the growth of L1210 leukemia, and the leukemic cell was poorly immunogenic in the mice. However, when OK-432 was given by ip injection in combination with mitomycin-C-treated L1210 cells, the growth of L1210 leukemia was significantly retarded and some mice did not "take" the leukemia. Meanwhile, BCG was also effective in this respect. Differences in properties as an immunopotentiator between OK-432 and BCG were suggested by the experiments of in vitro cytotoxicity test of spleen cells.


Cimetidine in anastomotic ulceration after partial gastrectomy. In order to assess the efficacy of cimetidine in patients with endoscopically anastomotic ulcer after partial gastrectomy, 21 such patients entered a double-blind prospective clinical trial. At endoscopy after 4 wk of treatment, 8 of 12 patients treated with 1 g of cimetidine daily compared with 1 of 9 patients who received a placebo had healed ulcers (P less than 0.05). Evaluation of symptoms relief supported the efficacy of cimetidine compared with placebo. Healing rate after 1 mo of treatment with cimetidine was 67% and increased to 86% after 2 mo. During 1 year of maintenance therapy with 800 mg cimetidine daily, 3 of 19 patients relapsed. No serious side effects were observed. The results of this study demonstrate a beneficial effect of cimetidine on healing and symptoms of anastomotic ulcers.


Relationship between membrane-bound immunoglobulin and viral antigens in liver cells from patients with hepatitis B virus infection. In an attempt to define further the significance of immunoglobulin G (IgG) fixed in vivo to the hepatocyte membrane in hepatitis B virus (HBV) infection, we have studied the relationship between presence of membrane-bound IgG and that of intracellular hepatitis B surface (HBsAg) and core (HBcAg) antigens in hepatocytes from 25 HBsAg chronic carriers. For this purpose, we have used a double immunofluorescence technique that is able to detect IgG and viral antigens within the same liver cell. In 15 patients with HBsAg-positive chronic active hepatitis, we found a statistically significant association between detection of membrane-bound IgG and that of intranuclear HBcAg within the same liver cells. On the contrary HGsAg containing hepatocytes generally did not show IgG fixed on their surface. IgG was not detected on the liver cell surface in 10 other HBsAg carriers without active disease and with large amounts of HBsAg containing hepatocytes. These results suggest that in HBsAg-positive chronic active hepatitis membrane-bound IgG is directed against viral antigens or virus-induced neoantigens that appear on the surface of infected cells at the time of active virus replication. Modulation of virus expression by this IgG could play a role in the pathogenesis of the disease.


Altered gastric emptying and secretion in primary anorexia nervosa. Primary anorexia nervosa (PAN) is an important psychiatric disease with a 7--21% mortality rate. Although altered gastrointestinal function may be an important aspect of its pathophysiology, no information is available concerning gastric emptying and secretion in those patients. During fasting, fractional emptying rates and hydrogen ion (H+) output were decreased twofold in PAN, as compared with healthy controls, and fluid output was slightly but not significantly decreased. Pentagastrin-induced peak stimulation of H+ output in PAN was 64% of that found in controls (P less than 0.05). Peak gastric fluid output was also significantly less in PAN patients, but suppression of fractional emptying produced by pentagastrin was of the same magnitude in both groups. Following a 250-ml water load, the magnitude and the duration of both the emptying and secretory responses were less in PAN patients than in controls. As a result, the initial increase of intragastric volume was greater in PAN patients than in controls, and the gradual return to fasting volume was delayed in those patients. Follow weight gain, fractional emptying tended to return toward control values, but was still significantly less than in controls following the water load. Gastric H+ and fluid output were not significantly modified following weight gain.


Fifteen years of clinical liver transplantation. Liver transplantation in humans was first attempted more than 15 yr ago. The 1-yr survival has slowly improved until it has now reached about 50%. In our experience, 46 patients have lived for at least 1 yr, with the longest survival being 9 yr. The high acute mortality in early trials was due in many cases to technical and management errors and to the use of damaged organs. With elimination of such factors, survival increased. Further improvements will depend upon better immunosuppression. Orthotopic liver transplantation (liver replacement) is the preferred operation in most cases, but placement of an extra liver (auxiliary transplantation) may have a role under special circumstances.


Functional interactions beween the DNA ligase of Escherichia coli and components of the DNA metabolic apparatus of T4 bacteriophage. T4 phage completely defective in both gene 30 (DNA ligase) and the rII gene (function unknown) require at least normal levels of host-derived DNA ligase (E. coli lig gene) for growth. Viable E. coli mutant strains that harbor less than 5% of the wild-type level of bacterial ligase do not support growth of T4 doubly defective in genes 30 and rII (T4 30- rII- mutants). We describe here two classes of secondary phage mutations that permit the growth of T4 30- rII- phage on ligase-defective hosts. One class mapped in T4 gene su30 (Krylov 1972) and improved T4 30- rII- phage growth on all E. coli strains, but to varying degrees that depended on levels of residual host ligase. Another class mapped in T4 gene 32 (helix-destabilizing protein) and improved growth specifically on a host carrying the lig2 mutation, but not on a host carrying another lig- lesion (lig4). Two conclusions are drawn from the work: (1) the role of DNA ligase in essential DNA metabolic processes in T4-infected E. coli is catalytic rather than stoichiometric, and (2) the E. coli DNA ligase is capable of specific functional interactions with components of the T4 DNA replication and/or repair apparatus.


Localized mutagenesis for the isolation of temperature-sensitive mutants of Escherichia coli affected in protein synthesis. Two variations of the method of localized mutagenesis were used to introduce mutations into the 72 min region of the Escherichia coli chromosome. Twenty temperature-sensitive mutants, with linkage to markers in this region, have been examined. Each strain showed an inhibition of growth in liquid medium at 44 degrees, and 19 of the mutants lost viability upon prolonged incubation at this temperature. A reduction in the rate of in vivo RNA and protein synthesis was observed for each mutant at 44 degrees, relative to a control strain. Eleven of the mutants were altered in growth sensitivity or resistance to one or more of three ribosomal antibiotics. The incomplete assembly of ribosomal subunits was detected in nine strains grown at 44 degrees. The characteristics of these mutants suggest that many of them are altered in genes for translational or transcriptional components, consistent with the clustering of these genes at this chromosomal locus.


Overlap hybridization screening: isolation and characterization of overlapping DNA fragments surrounding the leu2 gene on yeast chromosome III. A set of four plasmids containing overlapping segments comprising a total of about 30 kbp of cloned DNA from chromosome III of yeast (Saccharomyces cerevisiae) has been isolated and characterized by restriction endonuclease analyses and DNA:DNA hybridizations. Colony hybridization was carried out with labeled pYe(leu2)10, a plasmid carrying the yeast leu2 gene, to a bank of bacterial colonies containing recombinant plasmids constructed from the vector ColE1 and random fragments of yeast DNA. This resulted in the detection of two plasmids, pYe11G4 and pYe40C3, with DNA inserts which partially overlap the original cloned segment and contain additional DNA extending in opposite directions on the chromosome. By carrying out a second round of colony hybridization with pYe40C3, the cloned region was further extended in one direction. A region of DNA that is repeated at least ten times in the yeast genome was identified by hybridization of pYe11G4 to an EcoRI digest of total yeast DNA. The procedure described in this paper should allow the isolation of large sections of chromosomes, including non-transcribed regions, surrounding cloned genes.


Construction of an M13 histidine-transducing phage: a single-stranded cloning vehicle with one EcoRI site. In order to create a ready source of single-stranded DNA for DNA sequence determination by the dideoxy chain-termination method, the promoter-proximal part of the histidine operon, the hisOGD region of Salmonella typhimurium, was cloned onto the single-stranded phage M13. Both orientations of the his DNA were cloned to supply DNA template for sequencing of each strand. Insertion was achieved at an HaeIII site in the intergenic region (IR) of M13, and a single EcoRI site was purposely regenerated at one boundary of the his DNA insert. Infected colonies, not plaques, were selected using the hisD gene as a selective marker. The single RI site and the hisD marker for auxotrophic selection represent improvements on the wild type M13 as a single-stranded vector for cloning other DNA.


Mapping of restriction sites in the argF gene of Escherichia coli by partial endonuclease digestion of end-labeled DNA. A detailed physical map depicting the cleavage sites generated by ten different restriction endonucleases was prepared for the argF region of the Escherichia coli K-12 genome carried on a 1650 base pair fragment capable of directing the in vitro synthesis of ornithine transcarbamylase (OTCase; ec 2.1.3.3) under the control of arginine holorepressor. The method employed was originally developed by Smith and Birnstiel (1976), and involved the electrophoretic sizing of partial endonuclease digestion products of DNA radiolabeled at one end. This novel technique proved to be rapid, simple, amenable to the simultaneous mapping of numerous cleavage sites, and provided the essential information for determining the map order of restriction fragments. A facile method which involved magnesium phosphate as the DNA-binding agent was presented for the isolation of DNA fragments. The discovery of a 117 base pair leader sequence in the argF gene is also discussed.


How to manage adult respiratory distress syndrome. The key clinical features of adult respiratory distress syndrome are increasing dyspnea, tachypnea, and work of breathing; diffuse pulmonary infiltrations on chest radiographs; severe hypoxemia; and absence of a classic diagnosis. Adequate tissue oxygenation is the cornerstone of therapy. Therapeutic modalities include mechanical ventilation, fluid restriction, diuretics, and cardiotonic and vasopressor agents.


Leg ulcers in the elderly.

Leg ulcers in the elderly. The most common causes of leg ulcers in elderly patients are arteriosclerosis obliterans, venous insufficiency, hypertension, and prolonged pressure over relatively small body areas. The goal of treatment is to clear infection and stimulate good granulation tissue that can accept either a skin graft or epithelial regrowth from the borders of the ulcer.


Can drugs improve memory?

Can drugs improve memory? Drug-induced manipulation of the cholinergic system may possibly be able to reverse memory deficits due to old age.


Measuring intraepithelial lymphocytes, surface area, and volume of lamina propria in the jejunal mucosa of coeliac patients. An accurate morphometric method is described for quantifying the intraepithelial lymphocytes (IEL), the area of the epithelium, and the volume of the lamina propria in jejunal mucosa of coeliac patients. All measurements are related to a unit-length of muscularis mucosa which is unaltered by the disease process. The results show a significant decrease in the epithelial surface area and an increase in the volume of the lamina propria in coeliac jejunal mucosa compared with normal levels, even after treatment. The number of IEL is the same as normal before or after treatment. Other workers have shown an apparent increase in IEL in untreated disease which returns to normal levels after therapy. This discordance is explained and the importance of accurate quantitative methodology is stressed.


[Treatment of infected pseudoarthroses with electrodynamic field therapy]. The electrodynamic field treatment, its method and fundamentals in the therapy of infected, atrophic pseudarthrosis are described. In addition to the common principles of treatment, particularly the transplantation of autologous spongious bone is of great importance. The bone graft reestablishes normal electrical conduction within the pseudarthrosis and as a reactive receptor for the applied a.c.-potentials consolidates the defect. Up to now in 8 years of employing this method 271 patients, suffering from pseudarthroses have been treated, 91 of them had an infected, often atrophic pseudarthrosis. In each group in 92% of the cases bony healing could be achieved. Some case reports demonstrate clinic and therapy.


[Utilization of a synthetic skin substitute instead of split skin in cross-leg-flap and other pedicle flap grafting]. The method using synthetic skin substitute Epigard as a long-term coverage for donor sites in cross-leg-flap- and other plastic surgery procedures is described. The double-layered polyurethane/Teflon-material can be left on the wound for several weeks to avoid bacterial contamination during holding periods and to prepare regrafting of skin flaps. Experiences based on 32 cases are described.


On-line searches net data for administrators. New data base of references on health care administration and planning can be used at 900 centers, including many hospitals. Details on using the system are presented.


Who really likes HMOs?

Who really likes HMOs? The growth of HMOs has been slow, mainly because of a lack of acceptance by doctors and patients. Thus, the "swing" toward HMOs is likely to continue to proceed at a stately pace.


Corman loyal to universal NHI program.

Corman loyal to universal NHI program. Rep. James C. Corman has swung his support to the President's plan for national health insurance but still prefers universal public health care coverage.


Babesia infections in man.

Babesia infections in man. Only recently has it been recognized that hemotropic animal parasites of the genus babesia are also human pathogens. Similar to malaria in both symptoms and laboratory findings, acute babesiosis generally results in self-limited illness, but it can be fatal in the asplenic patient. The widely disseminated vector tick, Ixodes dammini, can transmit infection at each stage of its development, larva and nymph as well as adult.


The structural proteins and glycoproteins of herpesviruses: a review. The virions of different herpesviruses are similar with respect to the number and kinds of constituent polypeptides, in spite of variability in the structures of individual polypeptides. The total number of virion polypeptides and glycopeptides ranges from 20 to 30 for different viruses and, in general, no more than one-quarter of these polypeptides is detectable in naked nucleocapsids, implying that most of the virion polypeptides are acquired during the process of envelopment. Although the functions of most individual structural proteins have not been identified, one can predict that the nucleocapsid proteins serve primarily structural roles or may mediate packaging of the viral genome, that the non-glycosylated envelope proteins play essential roles in the process of envelopment and that the glycoproteins, which are probably all exposed to the virion surface, mediate adsorption to and penetration of the host cell. Two of the herpes simplex virus glycoproteins have been identified as targets of neutralizing antibodies and one of these proteins has been shown to mediate viral penetration, probably by promoting fusion between the virion envelope and cell surface membrane.


Collection of lymph from single lymph nodes and the intestines of fetal lambs in utero. Techniques for establishing chronic lymphatic fistulae in the intestinal and prescapular lymphatic ducts of fetal lambs during the test third of gestation are described. Despite the absence of antibody, immunoglobulin and extrinsic antigen, the number of recirculating lymphocytes increases considerably through gestation. Both thymus-derived and surface Ig-bearing B lymphocytes are present in the fetal recirculating pool and they appear to increase in number at the same rate.


B cell activity in rat bone marrow and spleen during primary and secondary response: dependence on strength of antigen. B cell activity was studied in the spleen and the bone marrow of inbred rats after applying xenogeneic erythrocytes of different antigenic strength (mouse or sheep). The hemolytic plaque technique of Cunningham was used. In contrast to the studies of other authors with other species, these experiments showed that in the rat the kinetics of IgM and IgG responses in bone marrow and spleen are principally similar during a primary as well as during a secondary reaction, varying only with the strength of the antigen.


Cocaine: magical drug or menace?

Cocaine: magical drug or menace? Cocaine has attracted much attention as a recreational drug and some advocates have acclaimed it as both pleasurable and safe. This paper reviews the available evidence concerning the drug's physiological and psychological safety. Few deaths are attributed to cocaine, but its use is presently restricted by very limited supplies. Use of the drug may possibly result in liver and respiratory problems and may lead to paranoid psychotic conditions. There are many deficiencies in our knowledge of the effects of this drug.


Fluorescence microscopy for detection of M. leprae in tissue sections. The fluorescence method was compared with the Fite-Faraco method for detecting acid-fast microorganisms in paraffin sections of cases of leprosy. Biopsies were obtained from 50 cases of leprosy covering all varieties and at varying stages of treatment. The fluorescence method was better than the Fite-Faraco method; 22 biopsies showing acid-fast organisms in fluorescence microscopy and 20 in the Fite-Faraco method. Its superiority was evidenced in two cases in which the organisms were very scanty. Fluorescence microscopy can also be used to determine the Bacterial Index and the Morphologic Index of organisms. The Morphologic Index, however, was one and a half times higher than that obtained by the Fite-Faraco technic. The ease and speed of fluorescence microscopy appear to be a great advantage.


Disposition and antiarrhythmic effect of lorcainide. The disposition and the antiarrhythmic effect of lorcainide (R 15,889) were investigated in 11 patients with ventricular premature beats (VPB) after a single intravenous dose of 100 mg or 2 mg/kg and after multiple intravenous and oral dosing. Pharmacokinetic parameters were computed according to the two-compartment open model. The half-life of the initial phase, t 1/2 (alpha), was calculated as 0.3 +/- 0.1 hr (mean +/- SD) and the terminal half-life, t 1/2 (beta), varied independently of the dose and route of administration between 5.8 and 12.5 hr (7.8 +/- 2.5 hr). After the single intravenous dose total plasma clearance (Cl) ranged from 570 to 1670 ml/min (988 +/- 425 ml/min) while after multiple dosing Cl decreased to 666 +/- 27 ml/min. Comparison of the area under the curves during steady state (ss) indicated a complete bioavailability of multiple oral doses. After the single intravenous dose, VPB were diminished or reduced for about 4 hr if the plasma concentrations exceeded 120 to 150 ng/ml. During ss-therapy plasma levels fluctuated between 200 and 550 ng/ml with an effective prevention of arrhythmias. Thus, the new drug demonstrates a therapeutic range of approximately 150 to 400 ng/ml and oral therapy seems to be effective with 100 mg t. i. d.


NAB 365 (clenbuterol) and salbutamol in asthmatics: a double-blind clinical trial. A double-blind comparison between orally administered NAB 365 (clenbuterol) and salbutamol was conducted in 30 impatients with either asthma or chronic bronchitis with asthma who had at least 15% reversibility in airway obstruction following inhalation of 1,500 microgram of orciprenaline. NAB 365 (clenbuterol) was administered to 15 patients at 30 microgram b.i.d. for 3 days; then the dosage was reduced to 20 microgram b.i.d. Salbutamol was administered to the other 15 patients at 4 mg t.i.d. Both drugs were administered for an average of 13 days. NAB 365 (clenbuterol) was shown to be an effective bronchodilator, with more rapid activity than that of salbutamol, on the FVC, FEV1, PEFR, the differences being significant on the third and seventh day of treatment. These findings are briefly discussed. No significant cardiovascular effects were noted for either drug, nor were differences found in need for additional treatment or in side-effects.


Hyperthermic inactivation of diploid yeast and the interaction of damage caused by hyperthermia and ionizing radiation. Inactivation of diploid yeast by hyperthermia has been studied. DO and Dq decrease with temperature for euoxic and anoxic conditions. The Arrhenius plot shows a break at 52 degrees C; the inactivation energies above and below this temperature are 153 and 94kcal/mol respectively. Hyperthermia (20 min at 51 degrees C) also potentiates the lethal action of gamma rays in diploid yeast cells under both euoxic and anoxic conditions. The interaction between hyperthermic and radiation damage appears to be largely at the sublethal level. The euoxic cells, the hyperthermic potentiation decreases with increasing time between the application of hyperthermia and radiation, being completely lost after 24 hours. However, in the anoxic cells there was no decrease in the hyperthermic potentiation with increasing time interval. These results suggest that yeast cells are capable of repairing hyperthermic sublethal damage, but require oxygen to do so. Thus there is a similarity in the process of repair of sublethal damage caused by ionizing radiation on the one hand and hyperthermia on the other.


Diagnostic evaluation of the patient suspected of having glaucoma. From the preceding it can be seen that the clinician still has little to help him decide which of his patients with elevated intraocular pressure will advance to frank glaucomatous disease. There are no objective provocative tests that will reliably predict the future. Consequently, one must obtain as much data as possible about an individual patient and, combining all these facts, have either a high or low degree of suspicion. Once a patient is noted to have elevated pressure, however, he must remain a glaucoma suspect, perhaps for the rest of his life. Only the subsequent examination will determine if glaucoma is continuing to be absent (or clinically undetectable) or whether it has actually become clinically manifest. The need for periodic reevaluation is obvious. Careful documentation of the disc, field, and applanation intraocular pressure over a period of time remains the most important aspect of the diagnostic evaluation of the patient with elevated intraocular pressure.


Immunoglobulin in urine from ileal conduit and nephrostomy patients. Patients with ileal conduits frequently have bacteruria, and some have pyelonephritis with associated renal deterioration. We attempted to find a noninvasive technique that could be used to define the source of the bacteria in such patients. Neither the antibody-coated bacteria test nor the selective immunofluorescent antibody studies made this distinction. Quantitative immunoglobulin determinations performed on urine from ileal conduits containing either less or greater than 10(5) bacteria per ml showed IgA to be the predominant immunoglobulin. Urine from patients with permanent nephrostomy drainage who had known chronic upper urinary tract infection contained predominantly IgG. Inasmuch as several studies have shown that both normal urine and urine from patients with pyelonephritis contain more IgG than IgA, we conclude that the conduit added IgA to the urine. However, even this quantitative immunoglobulin technique is unable to identify the source of the bacteria. To do so would have required an invasive collection of upper tract urine. The occurrence of acute symptomatic pyelonephritis was rare during the 18 months of the study, but the urine of the one ileal conduit patient with acute pyelonephritis studied contained more IgG than IgA.


Modified technique for microsurgical vasovasostomy. A modified technique of two-layer microsurgical vasovasostomy is described which permits continuous inspection of the lumen. A stricture-free anastomosis was documented in experimental animals by mating and scanning electron microscopic studies. The success of this procedure is attributed to precise placement of fine mucosal sutures, minimal fibrosis, and a fluid-tight anastomosis.


An electron microscopic study of the interaction between vesical epitherlium and E. Coli. Interaction between rat bladder epithelium and Escherichia coli was observed using the transmission electron microscope. As a first step, the bacterial cell coat attaches to the cell coat of the luminal membrane. Then, the luminal membrane enfolds to envelope the bacterium and take it within the cytoplasm. The unit membranes covering the bacteria are destroyed and some of these bacteria are found to be deformed and probably dead in the cytoplasm. Finally, the superficial epithelial cells invaded by many bacteria are collapsed and desquamated into urine. However, these epithelia show a rather prompt recovery and form the normal three layers within 24 hr after inoculation. This experiment suggests one of the defense mechanisms of the vesical epithelium againts E. coli. It is likely that the onset of clinical cystitis may be prevented by the same mechanism during the occasional bacterial invasion of the bladder.


Orgotein in equine navicular disease: a double blind study. Fourteen horses (7 treated with orgotein and 7 treated with a placebo) with navicular disease were studied on a double blind basis. All 14 horses had clinical and radiographic evidence of navicular disease. Orgotein and the placebo were administered by juxtabursal injection. Of the 7 orgotein-treated horses, 3 responded but none of the 7 placebo-treated horses responded. The difference was statistically significant (P less than 0.05).


Quantitative study on anomeric forms of glucose produced by alpha-glucosidases. Anomeric forms of glucose produced from phenyl alpha-maltoside, maltose, or phenyl alpha-glucoside have been determined quantitatively by simultaneous measurements of optical rotation and reducing power, for eight kinds of glucose-producing 1,4-alpha-glucosyl hydrolases, including glucose-forming amylase from human urine, and alpha-glucosidases from pig serum, honey bee, buckwheat seed, rice seed, sugar beet seed, flint corn seed, and brewer's yeast. All the eight enzymes studied were found to produce alpha-glucose exclusively.


Studies on bacterial chemotaxis. II. Effect of cheB and cheZ mutations on the methylation of methyl-accepting chemotaxis protein of Escherichia coli. Radioactive proteins from chemotactic mutants of Escherichia coli with continuous tumbling phenotype (cheB and cheZ) and their otherwise isogenic parent were compared by two-dimensional gel electrophoresis. The system was capable of separating non-methylated methyl-accepting chemotaxis protein (MCP) from its methylated equivalent. The analysis of proteins from the envelope fraction of the bacteria showed that the cheB mutants contained a larger portion of methylated MCP than did the parent. However, the change of MCP methylation level was small, if any, in cheZ strains. The results suggest that the product of cheB gene and the product of cheZ gene are not functional complementary. The product of cheB gene functions in controlling the level of methylation at the stationary state of the organisms. In addition to known MCP species, a new MCP of about 43,000 daltons was found. This MCP appears to be involved in transducing signals of some sugars.


Interaction of mitochondrial aspartate aminotransferase with negatively charged lecithin liposomes. Several kinds of hydrophilic proteins were examined to determine their interaction with artificial liposomes. Mitochondrial aspartate aminotransferase (m-GOT) [EC 2.6.1.1], as well as cytochrome c, was found to interact strongly with negatively charged liposomes. In each case, an appreciable amount of the protein bound to liposomes remained unreleased after raising the salt concentration in the medium. The m-GOT tightly bound to the liposomes was also found to become latent in its enzymatic activity, and could be reversibly activated by solubilization of the liposomes with detergent. This is also the case for cytochrome c, which ceases to be reducible by external reductant, such as dithionite. Furthermore, the tightly bound m-GOT was not susceptible to the proteolytic action of trypsin, or that of Nagarse. From these observations it can be inferred that these basic proteins interact with acidic liposomes not only electrostatically but also hydrophobically. This kind of hydrophobic interaction was not observed in the combination of positively charged liposomes and acidic proteins, including s-GOT. Mitochondrial GOT was shown to be bound to isolated intact mitochondrial, but the bound enzyme was fully active, in contrast to the case of acidic liposomes. The hydrophobic interaction of water-soluble protein with liposomes is discussed in connection with the penetration of matrix enzyme through mitochondrial membranes.