Idiopathic hemochromatosis. Demonstration of recessive transmission and early detection by family HLA typing. We studied iron overloading and HLA types in 24 sibships of patients with idiopathic hemochromatosis, of which 15 had at least two subjects with overt forms. HLA types of 84 unrelated patients were also investigated. Among siblings there was a significant association (P less than 0.0001) between the presence of hemochromatosis and the possession of the same two HLA haplotypes. The fact that overt forms of hemochromatosis depend on the presence of two specific homologous chromosomes strongly supports a recessive mode of transmission for the overt disease. The haplotypic equilibrium demonstrated in the unrelated patients group is another supporting argument. The lod-score value (2.239 for theta = 0.005) in six families available for study further supports the conclusion that a hemochromatosis gene is closely linked to the HLA-A locus. HLA typing in families with hemochromatosis could provide a means of early detection of subjects at risk before appearance of any sign of iron overload.


Functional capabilities of steroid-recruited neutrophils harvested for clinical transfusion. To determine whether steroids exert a direct inhibitory effect on neutrophil function, thus contraindicating their use to increase granulocyte yields for white-cell transfusions to infected neutropenic patients, we gave normal donors a single intravenous dose of dexamethasone (4 mg per square meter of body-surface area). Approximately two hours later the absolute neutrophil count increased from an initial value (mean +/- 1S.D.) of 3800 +/- 1400 to 5700 +/- 2400 per microliter. Granulocytes were then collected by discontinuous-flow centrifugation and tested. Viability (98.4 per cent), the percentage of neutrophils capable of phagocytosis (97.6), particle accumulation (12.3 candida per phagocyte), fungicidal activity (1.08 "ghost" yeast cells per phagocyte), the percentage of bacteria killed (97.7) and chemotaxis (119 per cent) did not significantly differ from observations in nontreated paired controls. The functional competence of neutrophils used for granulocyte transfusions was not altered after short-term exposure of these healthy donors to steroid medication.


Genetics of juvenile diabetes mellitus. A recessive gene closely linked to HLA D and with 50 per cent penetrance. We investigated the genetic predisposition to juvenile diabetes in the families of 31 index cases in relation to the inheritance of the HLA system. The diabetes-predisposing gene was found to be recessive because the diabetic sibs in index cases shared both their HLA genes with a significantly increased frequency. Penetrance was estimated at 50 per cent because half the HLA-identical sibs in index cases were diabetic. These conclusions fit with published observations that the risk to sibs of patients is about 100 per cent, when both parents are normal. In three informative cases of recombination within HLA the predisposing gene traveled with the HLA D segment of the recombinant haplotype. We prepared tables for the computation of risks to relatives, based on the hypothesis of recessivity, HLA linkage and 50 per cent penetrance.


Identification of a nondeletion defect in alpha-thalassemia. The molecular defect that has been demonstrated in alpha-thalassemia is the deletion of the alpha-globin structural genes. Since thalassemias are composed of heterogeneous groups of disorders, other types of defects could also result in alpha-thalassemia. We studied a Chinese family in which analysis of the mode of inheritance of alpha-thalassemia-1 and hemoglobin-H disease suggests a lesion that is not due to structural-gene deletion. Molecular hybridization studies with synthetic radioactive DNA's complementary to alpha-globin mRNA sequences show that in addition to the usual deletion defect, a nondeletion defect produces the phenotype of alpha-thalassemia-1. The combination of the deletion and non-deletion defects results in hemoglobin-H disease and not homozygous alpha-thalassemia associated with hydrops fetalis.


Evidence favoring the use of anticoagulants in the hospital phase of acute myocardial infarction. Since the last comprehensive review of anticoagulation in acute myocardial infarction four additional randomized control trials have been reported. The overwhelming majority of all trials favored anticoagulation. Rates of thromboembolism were higher in the control, and hemorrhagic complications in the anticoagulated group. Pooling of all randomized control trials gives mean case fatality rates of 19.6% for the control and 15.4% for the anticoagulated group, a relative reduction of 21% (P less than 0.05 or less than 0.001, depending on the analytic method). Five of six randomized control trials reported "no effect" because the difference favoring anticoagulation was not statistically significant. However, sample sizes in these "negative" papers were too small to protect against missing a 21% reduction in true case fatality rate due to anticoagulation (beta greater than 0.10). All patients who present no specific contraindication should receive anticoagulants during hospitalization for infarction.


Three-dimensional model of membrane-bound ribosomes obtained by electron microscopy. A low-resolution three-dimensional man has been obtained from crystalline arrays of membrane-bound eukaryotic ribosomes. It shows both ribosomal subunits to be adjacent to the membrane surface, attached to it by a part protruding from the large subunit.


Evidence for a sensitising pigment in fly photoreceptors. Many photoreceptor cells in invertebrates have a dual-peak spectral sensitivity. Evidence is presented that in fly photoreceptors the ultraviolet peak is due to a photostable pigment that absorbs light quanta and transfers the energy to the blue-absorbing visual pigment.


Nucleotide sequences of 5'-terminal ribosome-protected initiation regions from two reovirus messages. Sequences for two reovirus ribosome-protected fragments are presented. Comparison with four other reovirus initiation sites reveals only two common features: the 5'-terminal sequence m7GpppGmCUA, and (located 15-33 nucleotides from the cap) the sequence AUGG. Both the cap and the AUG codon are included in the 40S-ribosome protected region from all six reovirus messages.


Neurometabolic and behavioural effects of haloperidol in relation to drug levels in serum and brain. A  method has been developed for the quantitative determination of haloperidol in brain and other tissues. Such determinations have been made after acute and chronic administration of haloperidol to Sprague-Dawley rats. Different regions of the brain including the striatum, the limbic forebrain and the cerebellum have been analyzed separately. The haloperidol effects on Dopa formation have been studied in the same tissue samples. The stimulation of prolactin secretion via blockade of hypothalamic dopaminergic mechanisms and behavioural effects of the drug have been evaluated in parallel experiments. The elimination of haloperidol from brain tissue is a multiphasic process. The fourth phase of elimination is the slowest with a half life of 4 days. No strict correlation was found between serum and brain concentrations of haloperidol. Both after acute and chronic administration there exists apparently a saturating dose above which the brain concentration of the drug increases very little. The dose seems to coincide with that beyond which little increase in Dopa formation is observed. A pharmacokinetic analysis suggests an element of saturable binding or transfer of haloperidol to brain tissue. This mechanism is not preferentially localized to areas of brain rich in dopaminergic synapses. A good correlation was found between the haloperidol concentration in the brain on the one hand and its effects on behaviour, on serum prolactin values and on Dopa formation on the other.


Inhibition of acetylcholine turnover in rat hippocampus by intraseptal injections of beta-endorphin and morphine. Intraseptal administration of morphine (70 nmol) or beta-endorphin (0.7 nmol) reduced the rate of acetylcholine (ACh) turnover (TRACh) in rat hippocampus but not in striatum or cortex. These intraseptal injections failed to modify the ACh content and did not elicit analgesia. Naltrexone (15 mumol/kg, i.p.) completely antagonized the decrease of hippocampal TRACh elicited by the two opiate receptor agonists. Furthermore, intraseptal injections of naltrexone partially blocked the decrease in hippocampal TRACh induced by intraperitoneal administration of morphine (70 mumol/kg, i.p.). These data suggest that opiate agonists decrease hippocampal TRACh by regulating septal cholinergic neurons, and that this effect is not associated with analgesia.


Influence of enkephalin on K+-evoked efflux of putative neurotransmitters in rat brain. Selective inhibition of acetylcholine and dopamine release. In rat brain slices preincubated with various radiolabelled putative neurotransmitters, methionine-enkephalin diminished the potassium-evoked release of dopamine and acetylcholine. The effect was antagonised by naloxone. The potassium-induced effux of three other neurotransmitters, histamine, 5-hydroxy-tryptamine and gamma-aminobutyric acid, were unaffected by methionine-enkephalin. A probable physiological function for the endogenous ligands in specifically affecting the catecholaminergic and cholinergic transmission is suggested.


Behavioural and biochemical effects of chronic reduction of cerebral noradrenaline receptor stimulation. Mice were given various 7-day treatments designed to reduce cerebral noradrenergic function. Following these treatments, alterations in the sensitivity of limbic forebrain adenylate cyclase to noradrenaline (NA), and in the locomotor response of the mice to dopaminergic and noradrenergic agonists were studied. An enhanced response of the NA-sensitive adenylate cyclase to 1-NA(10-5 M), in comparison to control mice, was demonstrated after treatments producing a chronic reduction in stimulation of alpha-adrenergic receptors (phenoxybenzamine 10 mg/kg per day), beta-adrenergic receptors (propranolol, 0.05% diet), all noradrenergic receptors (reserpine 5 mg/kg, followed by FLA-63, 25 mg/kg per day), and dopaminergic as well as noradrenergic receptors (chlorpromazine, 5 mg/kg per day). Sensitivity of the NA-stimulated adenylate cyclase was not altered by chronic treatment with FLA-63 alone as a 0.05% diet. However, none of the chronic treatments except chlorpromazine administration increased the locomotor activity recorded for 2 h following apomorphine (1 mg/kg, s.c.) and none of the treatments altered the locomotor activity recorded in the 2 h following administration of the alpha-adrenergic agonist clonidine (1mg/kg, i.p.), or a combination of apomorphine (1 mg/kg, s.c.) plus clonidine (1 mg/kg, i.p.) The results support the hypothesis that noradrenergic stimulation plays a role secondary to that of dopamine in the production of locomotor activity.


Effects of pentoxifylline on red blood cell deformability and blood viscosity under hyperosmolar conditions. Using a hyperosmolar erythrocyte model, the authors studied the effect of changes in red cell deformability on whole blood viscosity and investigated the possibility of using pentoxifylline to modify red cell deformability and whole blood viscosity. In vitro studies reveal a significant correlation between the two parameters in as much as they are inversely proportionate, i.e. viscosity increases as red cell deformability decreases and vice versa. Addition of pentoxyfylline improves impaired red cell deformability under hyperosmolar conditions and simultaneously produces a reduction in whole blood viscosity.


[The development of cerebro-spinal fluid pathway in human embryos (author's transl)]. The early development of the subarachnoid space, the choroid plexus, and the arachnoid villi was studied in 60 normal human embryos ranging from Carnegie stage 12 to 23. The embryos were fixed in Bouin's fluid, paraffin-embedded, serially sectioned and stained with hematoxylin-eosin and Azan. One abnormal human embryo with exencephaly and myeloschisis in the high cervical cord was added for the study. A primitive subarachnoid space (future subarachnoid space) is first distinguishable as cavity formation within the meninx primitiva in the areas ventral to the middle brain vesicle at stage 14. The development of the primitive subarachnoid space precedes the appearance of the choroid plexus. The primitive subarachnoid space appears earlier in the region ventral to the rhombencephalon than in the region posterior to the fourth ventricle. By stage 20, a primitive subarachnoid space almost completely surrounds the neural tube. A fairly-well developed primitive subarachnoid space was observed in the abnormal human embryo, in which the fourth ventricle was open to the amniotic cavity and the ventricular system was completely separated from the primitive subarachoid space. These findings imply that the extraventricular spread of fluid of choroid plexus origin is not an essential factors, and that probably it is not even an important factor, for the development of the subarachnoid space. The arachnoid villi dose not appear even at the end of the embryonal stage. Absorption of the cerebrospinal fluid in an embryo should be done by the way other than the arachnoid villi.


[Endocrinological evaluation of sellar and suprasellar tumor cases (the ninth report)--on the PRL levels in the CSF (author's transl)]. Prolactin (PRL) levels in the cerebrospinal fluid (CSF) and plasma of 25 patients with sellar and suprasellar tumors, i.e. 5 of PRL secreting adenoma, 5 of GH secreting adenoma, 7 of so-called non-functioning pituitary adenoma, 5 of tuberculum sellae meningioma and 3 of craniopharyngioma, were studied in relation to the size of suprasellar extension measured by our own method on pneumoencephalograms. Plasma and CSF samples were obtained simultaneously at the time of lumbar puncture for pneumoencephalography. 1) Five patients of PRL secreting adenoma, whose plasma PRL levels ranged from 530 to 4,000 ng/ml, showed high PRL levels over 50 ng/ml in the CSF. On the contrary, 20 patients of the other tumors, whose plasma PRL levels were lower than 30 ng/ml, showed low PRL levels in th4 CSF, being below the sensitivity limit in the majority. 2) In 3 out of 5 PRL secreting adenomas, the PRL level in the CSF was higher than the plasma level and they showed a fairly large extrasellar extension. However in the remaining 2 the PRL level in the CSF was lower than a tenth of that in the plasma, and these revealed relatively small suprasellar extension. From these facts the presence of PRL at a high level in the CSF seems to be specific in the PRL secreting adenomas, and there seems to be a relationship between the CSF-plasma ratio of PRL level and the extrasellar extension in these cases.


[Bilateral direct injury of oculomotor nerves in craniocerebral trauma-report of a case (author's transl)]. A 46-year-old housewife was admitted to the Kawasaki Hospital soon after traffic accident. She was struck in the right forehead. She was very slightly drowsy but well oriented. Both eyeballs were extremely abducted without any motility. The pupils were bilaterally dilated and equal in size with no response to light. There was a pronounced blephaloptosis on both sides. Another neurological abnormality was a slight right facial weakness. On skull films, a linear fracture in the right temporobasal region was noted but the views of the orbits were normal. A lumbal tap showed a mild subarachnoidal hemorrhage. Three weeks after the trauma, pneumoencephalography showed no abnormal finding. Throughout convalescence, no special treatment was given beyond medication. She improved steadily. Three years after injury, re-examination disclosed that lateral gaze was almost full to either side but upward and downward movements of both eyeballs were limited. In our opinion, the most likely cause of the disability in this case was bilateral direct damage of the oculomotor nerves near the petroclinoid ligament (Grüber).


[A case with bilateral internal carotid and left vertebral artery occlusion (author's transl)]. Since Moniz reported the obstruction of the extracranial internal carotid artery in 1937, there have been many reports of extracranial arterial occlusion. The authors experienced a case  of  40-year-old woman who had minor neurological deficit such as attack of black out, disturbance of the bilateral visual acuity, concentric narrowing of the bilateral field and disturbance of the bilateral auditory acuity, in spite of complete obstruction of both internal carotid arteries and the left vertebral artery, and possible obstruction of the right vertebral artery. The angiograms showed well developed collateral circulation. The cause of the obstruction was considered to be angitis following otitis media and meningitis in her childhood. The symptoms were alleviated by drugtherapy.


A case of intracranial osteochondroma (author's transl). The authors reported a case of intracranial osteochondroma. A 52-year-old woman was admitted to Tokyo Women's Medical College hospital with seven years history of right visual disturbance and frontal heavy feeling. Neurological examination revealed merely right optic nerve impairment. A large mottled calcification in X-ray craniogram and a localized homogenous high density area illuminated by EMI scan in the right parasellar region were characteristic in diagnosis of osteochondroma. On April 21, 1976 a right fronto-temporal craniotomy was performed and the tumor tissue was removed partially. The pathological examination confirmed the diagnosis of osteochondroma (ossifying chodroma).


[Intracerebellar Hematoma following microsurgical STA-cortical MCA bypass surgery (author's transl)]. Since October 1967 we have performed 76 microsurgical STA--cortical MCA bypass operations. Recently we have had two cases, who died from intracerebellar hematoma following bypass operation. Intracerebellar hematoma is reported primarily due to hypertension (50-80%), and to comprise 10% of all spontaneous intracranial hematomas. Diagnosis of this lesion is frequently missed but can be made by the typical clinical picture (respiratory irregularity, pinpoint pupils, absence of oculovestibular responses, loss of consciousness), and the CT-Scan. If the correct diagnosis is made and operation promptly performed, many patients with subacute or acute intracerebellar hematoma can be saved. The 2 cases presented here had a history of hypertension and anticoagulation (including Colfarit), but had sustained the bypass operation well and showed no neurological deficit immediately after the operation. They had received Rheomacrodex intra- and postoperatively. Quite soon postoperatively, however, the systolic blood pressure rose to 210 mmHg and the patients complained of severe headache. They were treated symptomatically with analgesics and antihypertensive drugs. A short time later they became comatose and died. In order to prevent this complication after bypass surgery, postoperative management of hypertension is mandatory. The combination of antithrombic agents, Colfarit and Rheomacrodox, might have played a role in inducing the hemorrhages. Furthermore strong analgesics should be withheld to prevent their masking neurological deterioration. Intracerebellar hematoma must always be considered in hypertensive or anticoagulated patients, especially because it can be cured with prompt diagnosis and operative treatment.


[A case of posterior fossa acute subdural hematoma diagnosed through extravasated contrast media (author's transl)]. The authors report a case of posterior fossa acute subdural hematoma diagnosed preoperatively through extravasted contrast media from the hemispheric branch of the posterior inferior cerebellar artery along a linear fracture across the transverse sinus unto the pyramis. In a review of the literature it is to be noted that in acute cases, nystagmus and cerebellar signs are very rare and patients fall in coma within 24 hours after injury with promptly advancing severe brain stem compression signs in contrast to subacute or chronic cases. Therefore, whenever the diagnosis is suspected, time should not be lost for a burr hole opening and decompressive craniectomy. Vertebral angiography is sometimes very confirmative but it should be done only when time affording.


Effects of food restriction on circadian adrenocortical rhythm in rats under constant lighting conditions. Influences of food restriction on the circadian adrenocortical rhythm were studied in blinded and unblinded rats under constant light (LL). Blinded rats, which were allowed to access to food ad libitum, manifested a phase shift of the circadian adrenocortical rhythm. However, restricted feeding between 19.00 and 21.00 h for 4 weeks prevented the phase shift and the peak levels of plasma corticosterone (B) still appeared at 19.00. On the other hand, unblinded rats, allowed to feed ad libitum, showed an abolition of the circadian periodicity of adrenocortical activity 5 weeks after exposing the rats to LL. However, the rhythm, synchronizing with the feeding periods, was promptly recovered under food restriction. These results suggest a synchronizing and generating action of feeding for the circadian adrenocortical rhythm.


Electrophysiological and pharmacological studies of amygdalo-hypothalamic connections in the rat. Electrophysiological techniques have been used to investigate the neural connections between the amygdala and the rostral hypothalamus of female rats. Recordings have been made from 120 neurones in the preoptic/anterior hypothalamic area (POA/AHA) which could be driven by electrical stimulation of the amygdala. 89% of the neurones were activated orthodromically and, of these, 2 groups have been distinguished according to the latency of the responses. The remainder of identified cells were antidromically activated by stimulation of the amygdala. The response of these identified preoptic cells to the iontophoretic application of dopamine (DA) and noradrenaline (NA) has also been investigated. DA inhibited the spontaneous or glutamate-induced activity in 18 of 21 identified cells so tested and NA inhibited all 25 units; excitation was never observed. The role of the amygdala and of aminergic pathways in the control of ovulation is discussed.


Iontophoretically applied corticosteroids do not affect the firing of hippocampal neurons. In view of the possible role of the hippocampus in the neuroendocrine regulation of ACTH secretion, experiments were conducted in rats and cats in which cortisol (C) and corticosterone (CS) were applied by iontophoresis and pressure microinjection while recording from single identified pyramidal cells in the dorsal hippocampus, using multi-barrelled microelectrodes. None of the more than 500 cells tested responded with a significant change in firing rate. The possible relation of these findings to neuroendocrine regulation of ACTH secretion is discussed.


The angiographic diagnosis of occlusions of the posterior cerebral artery. The angiographic appearance of twenty-seven unilateral or bilateral occlusions of the posterior cerebral artery (PCA) are described. They are classified into proximal, interpeduncular, ambient or distal occlusions according to their prefered location. Ambient occlusions were by far the most common and have therefore probably not developed from a stenosing arteriosclerosis since this forms preferentially in the proximal and interpeduncular segment. Leptomeningeal anastomoses were common between all respective branches of the middle cerebral artery and the PCA but the anterior cerebral artery and the interarterial anastomoses rarely contributed to the collateral blood supply. A retrograde progression of the occlusion was observed on reangiography occasionally in cases in which there was no major artery originating just proximal to the occlusion. Anterograde progression and disintegration of obstructing material was also observed. PCA occlusions had a good prognosis in terms of survival but the associated visual field defects rarely improved.


Angiographic demonstration of lupus cerebral phlebitis with communicating hydrocephalus. Two patients with systemic lupus erythematosus, who had striking angiographic abnormalities of venous contour, are presented. Both cases also had communicating hydrocephalus. Postmortem examination of one patient led to a histological explanation of the venous changes observed radiologically. Previous radiological reports have shown involvement of arteries but not veins with lupus erythematosus. The angiographic signs of phlebitis may signify a meningeal reaction which, when combined with ventricular enlargement, should suggest communicating hydrocephalus.


The glabella-inion line as a baseline for CT scanning of the brain. The optimal position of the head for CT scanning of the brain is discussed according to the statistical data of roentgencraniometry. The glabella-inion (GI) line was shown to be parallel to the frontal-occipital pole (FO) line which was substituted for a line joining the deepest gyral impressions in frontal and occipital parts of the cranial cavity. On the other hand, Reid's baseline and the canthomeatal (CM) line intersected the GI line at various angles from person to person. There was a significant difference between each average inclination angle of males and females. Regarding using either Reid's or the CM line for a baseline, it should be noted that the brains were scanned in almost random direction. Therefore it may be convenient to apply the GI line for comparing the findings between individuals, and for correlating them to the anatomical diagnosis.


Computed-tomography of bilateral isodense chronic subdural hematomas. While unilateral chronic isodense subdural hematomas as a result of indirect signs of a space-occupying lesion are easily recognizable on computed tomography (CT) and clearly diagnosed on the angiogram, bilateral chronic isodense subdural hematomas may cause considerable difficulty. In two cases with CT false negative findings we observed, retrospectively, significant small cellae mediae and also the main part of the anterior horns sharply pointed and approaching one another. Three further cases showed the same ventricular configuration, which we called "hare's ears sign". This sign together with clinical data is always suspicious of chronic bilateral isodense subdural hematomas and carotid angiography is indicated. Other possible signs are: subtle midline shift if the size of the hematoma varies, changed formation of density of brain tissue, non-appearance of cerebral sulci especially in elderly patients, and eventually the visualization of a membrane after intrevenous injection of contrast material.


Experimental embolization with inflatable and releasable balloons in dogs. Experimental arterial occlusion was performed on eight dogs by means of inflatable and releasable balloons, in five of which a reliable and long lasting arterial occlusion was obtained. Three accidents occurred. One dog died when the liquid latex was emptied spontaneously from the balloon and the territory was embolized causing a severe general reaction. In two other dogs the emptying of the balloon, inflated with iodine, was followed by distal embolization of the empty latex balloon. The possibility of such accidents would be too great a risk for human application, but it now seems possible with the new glue, isobutyl-2-cyanoacrylate, to embolize vascular territories more accurately and more safely by means of inflatable and releasable balloons.


[Type B viral hepatitis. Recent findings]. The discovery of hepatitis B surface antigen by Blumberg in 1965 offered a specific marker that could readily be employed in the clinical, epidemiological and experimental investigation of type B viral hepatitis. It has since been followed by the continuous gathering of further knowledge concerning this disease. The morphology and immunological and biochemical features of virus B, the epidemiology and prevention of hepatitis B, and marker determination techniques are examined in an overview of the present situation.


[Bone lesions of the foot in diabetic disease]. Arthropathy is increasingly reported in diabetes. In a personal series of 300 subjects  with undoubted diabetes mellitus (mean age: 67 yr), five cases of arthropathy (1.6%) were noted. The pathogenesis of this association is discussed, particularly in the light of the concomitance of neuropathy in 4-5 cases. The main clinical signs included: swelling, redness, mal perforans, and pied cubique diabetique. The incubation period of arthropathic lesions is also examined. The medical and surgical treatment of diabetic arthropathy is described.


[Role of the family group in attempted suicide among the young]. Data from 1848 cases of attempted suicide on the files of the Milan University Emergency Psychiatry Ward and 990 suicides recorded by the Milan Legal Medicine Department in the period 1964-1969 were computerised on specially prepared cards, designed to permit objective study of acts of self-destruction in the light of the relations between the subject, his family, and the community. The results point to the importance of extra-subjective factors (loss of cohesion in the family, the way in which parents carry out their respective roles, various environmental factors) as motive forces in attempted suicide.


[Findings on serum anti-myocardium antibodies in heart disease. Preliminary communication]. The research of antiheart antibodies was carried out on 174 cardiac patients. The 40% of patients with acute myocardial infarction and the 27% with other heart diseases evidenced antimyocardial antibodies. No antipericardial antibodies were found. The immunological research seems to offer an important help in the diagnosis of the atypical postmyocardial infarction syndromes, of the recidivant myopericarditis without clinical or electrocardiographic evidence; and sometimes it may reveal subclinical myocardial infarction.


[Anthropometric state of nutrition in students of various elementary and secondary schools in Ancona]. An anthropometric assessment was made of the nutritional state of children from Ancona elementary and middle schools. It was found that the girls had more body fat than the boys, though the latter, too, displayed values above those reported in U.S. and Italian studies. The view is expressed that this excess fat requires counteraction through nutritional education. Diet and physical exercise are known to be important in the prevention of degenerative diseases of a metabolic nature. Such prevention must be undertaken as soon as possible.


[A bladder calculus of unusual dimensions]. A case of giant vesical calculosis is presented on account of its statistical and radiographical unusualness. Its aetiopathogenesis is briefly discussed.


[Principal clinical indications for direct transfusion therapy]. So-called "directed" transfusion results in better utilisation and hence a saving of blood, along with less risk to the patient. An examination is made of the few diseases in which the transfusion of whole blood is indicated, and the very many cases in which blood derivatives can be usefully employed.


[Thalassemia trait and the lithogenicity of bile]. The study of bile composition in thalassemic patients revealed a lithogenic bile even without gallstones. The lithogenic index was higher than in a group of colelithiasic non hemolytic patients. The possible correlation between lithogenic bile and congenital alteration of the lipidic metabolism in thalassemic patients is discussed.


[Cholesterol-7-alpha-hydroxylase activity in cholelithiasis]. The physiopathological mechanism of cholesterol supersaturation in the bile was investigated by determining cholesterol-7-alpha-hydroxylase activity in 5 normal controls and 8 subjects with cholesterol gallstones. Bile lipids were also evaluated. Hydroxylase values were higher (though not significantly) in the controls. A linear correlation (r = 0.797) between the hydroxylase and bile acid concentration was noted in both groups.


[Bile phospholipids; function and significance]. The part played by phospholipides in the genesis of cholesterol gallstone considered. This is present in patients who frequently present a lecithin synthesis defect at hepatic level since precursors are used for forming triglycerides. Nevertheless polyunsaturated phosphatidicholine has a negative influence on the SB + PL/C ratio in the bile of T-tube subjects receiving 2 g of substance i.v. for 5 days.


[Effect of chenic acid on bile lipid composition and on metabolism of bile acids in lithiasis patients]. Gallbladder bile unsaturated with cholesterol is a prerequisite for gallstone dissolution. Chenodeoxycholic (chenic) acid ingestion by patients with gallstones induces saturated bile to become unsaturated. Additionally, chenic acid becomes the predominant bile acid in bile and the chenic acid and total bile acid pools increase in a majority of such patients. Most importantly, the hourly secretion of cholesterol decreases during chenic therapy such that the cholesterol to bile acid ratio in bile is significantly lower and bile becomes unsaturated with cholesterol. Preliminary evidence suggests that the mechanism of action of chenic acid may relate to a suppression of the rate limiting hepatic enzyme for cholesterol synthesis, HMGCoA reductase.


[Changes in bile and blood lipid levels following chenodeoxycholic acid therapy]. Original and known methods and ordinary routine tests were used to evaluate changes in bile salts, phospholipids and cholesterol and in serum lipids induced in 50 patients during treatment with chenodeoxycholic acid (2-12 mg/Kg/day) for about 15 months. No significant differences in serum lipids, though a clear fall in triglycerides was noted in those who received 7-12 mg/Kg/day as opposed to 2-7 mg/Kg/day. There was a marked fall in the molar percentage of biliary cholesterol, followed by a significant reduction in the saturation index of Metzger. Once again, greater reduction was noted in patients receiving the higher dose range. A correlation between the dose used and the percentage of cholesterol in the bile was thus evident. After treatment, chenodeoxycholic acid was the main bile acid and there was a significant fall in cholic and deoxycholic acid. The desired drug effect can be obtained with low doses (7-12 mg/Kg/day) and a biliary chenodeoxycholic value well below 90%.


[Results of chenic acid therapy in patients with cholesterol calculi]. 58 patients suffering from cholesterol gallstones were treated with chenodesoxycholic acid at a dose of 2-15 mg/Kg/die for about 15 months so as to determine its effettiveness and establish a minimum effective dose. 8 of these patients presented stones in the common bile duct. The results showed: a complete dissolution of the stones in 16 of the 50 patients considered to be suffering from cholecystic lithiasis; partial dissolution in 15; no change in the remaining 19. Of the 8 patients with bile duct stones, 3 presented complete dissolution, 2 partial, and 3 no change. An improvement in digestive function was observed in most cases. The liver function tests carried out in each patient before, during and after treatment with chenodesoxycholic acid were completely normal.


[Correlation of malabsorption of bile acids, bile lipid composition and calculi]. The relation between malabsorption of bile acids, the bile lipid composition, and biliary stones was examined in 8 patients subjected to ileal resection (particularly for Crohn's disease), 6 with ileal bypass for morbid obesity, and 10 healthy controls. The 1-14C-cholylglycine breath test was employed to detect of the absorption and deconjugation of bile acids. Bile lipid composition was expressed according with Metzger's saturation index. Healthy subjects gave normal findings in all respects, whereas ileal resection was accompanied by malabsorption, increased deconjugation, and faecal loss of 14C. These changes, particularly malabsorption, were more evident after ileal bypass. Preoperative saturation values rose to more than 1 in all cases, especially after resection. Liver disease (steatosis and cirrhosis) 6 months after bypass, together with cholesterol lithiasis in 2/6 patients.


[Dynamic BSP tests in cholelithiasis patients before and during chenodeoxycholic acid therapy; initial results]. The BSP loading and continuous infusion tests were performed in 7 patients before and after a two-week treatment with chenodeoxycholic acid (10 mg/Kg/die per os). The following parameters were evaluated: 45th minute retention percentage, plasma disappearance rate during the first 16 minutes, compartmental transfer rates, transport maximum and storage. Our results indicated that no cholestatic effects seemed to be induced during the first period of treatment with chenodeoxycholic acid.


Elective appendectomy with abdominal and pelvic surgery. A series is presented of 830 patients in whom elective appendectomy was performed at the time of laparotomy for other intraabdominal disease. Special emphasis is given to 490 such procedures among 1042 patients with abdominal hysterectomy, an incidence of 47%. This increases to 57% by exclusion of patients with previous appendectomy. The contraindications to elective appendectomy are discussed, as well as its morbidity, mortality, and complications. The author concludes that an elective appendectomy should be performed with abdominal and pelvic surgery whenever the opportunity is presented, provided that the procedure is not prohibited by the contraindications discussed.


Ruptured tuboovarian abscess. Is hysterectomy necessary? The use of conservative pelvic surgery combined with intra- and postoperative antibiotic peritoneal lavage has been evaluated in 113 women with generalized peritonitis due to ruptured tuboovarian abscess. Mortality was 7.1% and hysterectomy was only required in 3% of cases. Hormonal and menstrual functions were retained in 73.5% and the potential for future pregnancy in 42.5%. Further surgery was required in 17.5% of the patients. These results are contrasted with recent series of comparable size in which the mortality and morbidity were similar but the frequency of hysterectomy ranged from 70--100%. It is concluded that hysterectomy is not necessary in the management of ruptured tuboovarian abscess if the major source of sepsis is removed and adequate peritoneal drainage is provided by the use of lavage.


Congenital absence of the uterus. Clinicopathologic and endocrine findings. Laparoscopic ovarian biopsies performed on 11 patients with congenital absence of the uterus revealed the presence of follicles at different stages of development, with corpora lutea in 2 patients and cystic follicles in 1 patient who had cyclic patterns of serum gonadotropins and plasma progesterone.


Clinicoepidemiologic study of uterine cancer. Comparative aspects of the endometrial and cervical sites. A review of all newly diagnosed cases of cervical and endometrial carcinoma in Israel during the 5-year period of 1961-1965 yielded mean annual incidence rates of 4.9/100,000 and 7.4/100,000, respectively. Cervical cancer was more prevalent in Moroccan-born women and among divorcees, while the risk of endometrial cancer was highest in older age, among the European born, and the single; it also appeared earlier in life. Postmenopausal bleeding constituted the most frequent first symptom in both sites. Fifty percent of the patients of both groups were diagnosed within 1 month, but the delay was somewhat longer in the endometrial group. Median survival was 5 years in patients with cervical cancer and above 12 years in those with cancer of the corpus. Five-year survival was 50 and 75%, respectively. Survival tended to be better in younger patients in both groups. It is expected that the gradual disappearance of intraethnic differences in Israel would lead to a decrease in the incidence of invasive cervical cancer, coupled with an increased incidence of the endometrial category.


Abdominal pregnancy. Review of current management. Two case reports of intraabdominal pregnancy and review of 9 others which have been seen at the Indiana University Hospital are presented. An incidence of one intraabdominal pregnancy in 7931 deliveries at Indiana University is compared to 1 in 3372 deliveries from a large series at another institution. A perinatal mortality of 91% and a maternal mortality of 18% from our series are contrasted to 90% and 6%, respectively, from the world literature. The diagnosis, management, and outcome of this rare form of extrauterine pregnancy are discussed. Recurrent abdominal pain in the gravid patient may signal abdominal pregnancy. The role of ultrasound in diagnosis and the importance of early surgical intervention with minimal disturbance of the placenta are stressed. In cases of intraabdominal pregnancy, the infant rarely survives and congenital malformations are frequent.


Clinical and endocrine studies in menopausal women after estradiol pellet implantation. Eight patients who required prolonged estrogen replacement therapy were implanted with two 25-mg pellets of estradiol-17beta and were followed with serial measurements of serum estradiol, estrone, follicle stimulating hormone, luteinizing hormone, and maturation index of the vaginal smears. There was a significant increase in estrogen levels within 24 hours after pellet implantation, and this level was maintained in physiologic premenopausal range during the rest of the period of followup. Gonadotropins were suppressed to a greater degree in gonadal dysgenesis patients as compared to postmenopausal patients. There was rapid relief of vasomotor symptoms within 4--10 days even though the gonadotropin levels were high. Implantation of 50 mg of estradiol pellets and periodic withdrawal bleeding with a progestational agent seems to be an effective method of long-term replacement therapy.


A rapid modification of the beta-hCG radioimmunoassay. Use as an aid in the diagnosis of ectopic pregnancy. Over half the patients with proven ectopic pregnancies have negative urinary slide or tube pregnancy tests at the time they present with symptoms. Because of the morbidity and mortality associated with unrecognized ectopic pregnancy, the need for an accurate, rapid, sensitive, and specific method for measuring low levels of human chorionic gonadotropin (hCG) is apparent. A three-hour radioimmunoassay (RIA) for serum hCG with a sensitivity of 5 mlU/ml was developed which utilizes an antiserum generated against the beta subunit of hCG, 125I-hCG for tracer purposes, a highly purified hCG standard, and dioxane for separation of bound from free hormone. Blood samples were obtained from 52 patients with suspected ectopic pregnancies. Twenty-four of these patients had a surgically proven ectopic gestation. The blood samples were analyzed in the above described RIA, and in the standard (24-hour) double antibody beta-hCG RIA. There were no false positive or false negative results by either assay. These results indicate that the dioxane modification of the beta-hCG RIA is an accurate and rapid method for confirming the diagnosis of ectopic pregnancy.


Serum prolactin in normal and pathologic pregnancy. Serum prolactin values in normal pregnant women showed a progressive increase from a mean value of 50 ng/ml in the 12th week to 270 ng/ml at term, with the range at term being 100-600 ng/ml. There was a fairly good correlation (r = 0.7) between the values of 24-hour urine estriol in 138 determinations and in the serum prolactin in 133 pregnant women. The regression lines of serum hPRL values with time of gestation in cases of intrauterine growth retardation (IUGR) and diabetes mellitus were less steep than those seen in normal pregnancy. The serum hPRL value of patients with preeclamptic toxemia, latent diabetes, premature rupture of membranes, or multiple pregnancies were found not to differ significantly from the values observed in normal pregnancy. The results indicate that prolactin determinations in pathologic pregnancies are not useful as an aid in their evaluation.


Intrapartum fetal cardiac arrest. A preliminary observation. In 13 patients, episodes of transient fetal cardiac arrest were observed in a group of 594 extensively monitored labors during a given 3-year period. The number of episodes per patient ranged from one to six, with a maximal duration of cardiac arrest (R-R interval) being 5.2 seconds. All of the patients responded to changing maternal position or termination of pregnancy except 1. This patient received Atropine as a premedication for cesarean section. The parasympatholytic properties of Atropine minimized the severity of cardiac arrest. The effect of cardiac arrest on fetuses is not clearly shown in these preliminary observations. The prompt elimination of cardiac arrest is thought to be imperative in reducing perinatal loss. Cardiac arrest is though to be an extensive form of severe variable deceleration. The hypothesis is made that these fetuses had an unbalanced autonomic nervous system and/or an overwhelming vagal tone. If these signs are detected early by fetal monitoring, attention should be paid to the possibility of cardiac arrest.


Analysis of heart rate patterns preceding fetal death. An analysis of fetal heart rate patterns preceding death is presented in 7 patients. The intrapartum deaths were not predictable on the basis of clinical evaluation. Predeath recording time varied from 11 minutes to 7.75 hours. An atypical deceleration pattern was observed in 3 patients terminally with associated loss of beat-to-beat variability. These occurred with uterine contractions, but had the characteristic appearance of variable decelerations (CC). This pattern was characterized by short contraction to deceleration intervals, and by large drops in the rate (62-102 beats/min). In 6 of the 7 patients, there was either absent (0-5 beats/min) or diminished (5-10 beats/min) beat-to-beat variability. In 1 the terminal recording showed an absolutely fixed rate with a progressive loss of beats and an increased amount of electrical noise.


Placental infarction leading to fetal or neonatal death. A prospective study. The pathogenesis of perinatal death due to placental infarction was analyzed in a large prospective study that included more than 1000 medical, demographic, hereditary, and postmortem variables. The disorder was the fifth most frequent cause of death in the study with a perinatal mortality of 2.26/1000 births. Its frequency was directly correlated with the gravidas' blood pressures, an effect augmented by albuminuria and work during pregnancy. Fatal infarcts were increased 20-fold with glomerulonephritis, fivefold with abruptio placentae, and twofold when the gravidas' hemoglobins were over 12 g/100 ml. Maternal weight gains were suboptimal and the involved neonates had a pattern of growth retardation characteristic of undernutrition. The disorder was more frequent when the gravida had made few prenatal visits for medical care and had had prior unsuccessful pregnancies.


Ultrasonic placental localization and bloody taps in midtrimester amniocentesis for prenatal genetic diagnosis. In a prospective study, a small team of obstetricians concurrently performed midtrimester amniocentesis for prenatal genetic diagnosis on 32 patients after ultrasonic placental localization and on 50 patients without prior ultrasound. The use of ultrasound did not affect the red cell count in the fluid samples. Furthermore, the 2 patient groups had similar rates of grossly bloody taps. The average number of viable clones after culture was lower both after bloody taps and when ultrasound had been used, but in neither case was the difference significant. There were no postamniocentesis complications in the entire patient population. These results suggest that ultrasonic placental localization is not helpful in avoiding bloody taps, and that bloody taps may not necessarily be dangerous. Therefore, since the long-range hazards of in utero exposure to ultrasound may not yet be known, it would seem judicious to use preamniocentesis ultrasound selectively and on specific indication, rather than routinely.


Cesarean section and maternal mortality in Rhode Island. Incidence and risk factors, 1965-1975. An 11-year study from January 1, 1965, through December 31, 1975, was undertaken to study maternal death associated with cesarean section in Rhode Island. The risk of maternal death associated with cesarean section and that for vaginal delivery was calculated, and it was found that the risk of death from cesarean section was 26 times greater than with vaginal delivery. The expanding indications for cesarean section, factors associated with excess risk of death from cesarean section, and suggestions to prevent such catastrophe are discussed.


Comparison of nitroprusside and hydralazine in hypertensive pregnant ewes. During phenylephrine-induced hypertension in 6 near-term pregnant ewes, nitroprusside rapidly lowered the blood pressure to control values with no accompanying change in uterine blood flow. Hydralazine slowly lowered the blood pressure to control values. This change was accompanied by a statistically significant (15%) increase in uterine blood flow. Hydralazine also had a more pronounced effect than nitroprusside on cardiac output, heart rate, and total peripheral resistance. There were no significant acid-base changes in mother or fetus throughout the study.


Effect of terbutaline on cardiovascular state and uterine blood flow in pregnant ewes. The cardiovascular and uterine hemodynamic effects of terbutaline, a beta-adrenergic receptor stimulant and labor inhibiting agent, were evaluated in the chronically instrumented, near-term pregnant ewe. The administration of terbutaline in the dose range required for labor inhibition in this species resulted in a mild maternal tachycardia and increase in pulse pressure without significant changes in uterine blood flow; uterine vascular resistance; or systolic, diastolic, or mean blood pressures. With infusion rates of terbutaline in excess of those required for labor inhibition, significant increases in maternal heart rate, pulse pressure, and systolic blood pressure were observed. Diastolic blood pressure decreased significantly during the higher infusion rates; however, uterine blood flow was unaffected. The minimal cardiovascular and uterine blood flow was unaffected. The minimal cardiovascular and uterine hemodynamic effects noted with the administration of terbutaline in the dose range necessary for labor inhibition indicate that this agent may possess advantages over several others currently in use for the treatment of premature labor.


Phenobarbital prophylaxis of neonatal hyperbilirubinemia. Evaluation of a nightly dose. In an attempt to improve the success of a previous study of the prevention of neonatal hyperbilirubinemia employing phenobarbital in a single 30-mg nightly dose from 32 weeks' gestation until delivery, 246 patients were tested against 270 control subjects. A 39% reduction in significant hyperbilirubinemia was realized but this fell far short of previous findings  with a  larger dose. Premature infants received no apparent protection against jaundice. Mature infants had no severe jaundice (more than 16 mg/100 ml) and the resolution of hyperbilirubinemia was more rapid. No apparent difference in drowsiness or daytime sedation from previous experience was found. Female newborns responded much better (3:1) to prophylaxis. These disappointing results favor a return to the 1-g daily divided dose.


A simple test for predicting pregnancy-induced hypertension. One hundred thirty-one nulliparous patients between 28 to 34 weeks' gestation with no known history of renal disease or hypertension were placed in the left lateral recumbent position and their blood pressures were taken once and recorded. They then turned to the supine position where another blood pressure was taken within 2 minutes and also recorded. All these patients were then followed routinely throught their pregnancy, labor, delivery, and postpartum period. Of those patients whose diastolic blood pressure during the modified "roll-over" test rose less than 15 mmHg, only 2 of 99 developed pregnancy-induced hypertension (PIH). Of those whose roll-over test showed an increase of more than 15 mmHg, 23 of 32 developed PIH (P less than .000000001). Of those whose blood pressure during the roll-over developed PIH(P less than .00000001). Also, 10 of 11 patients whose diastolic blood pressure increased more than 25 mmHg during the roll-over test developed PIH (P less than .000000001).


Experiences with a fine stainless stell wire fetal scalp electrode. A simple, economic stainless steel wire fetal scalp electrode is described. Its uneventful use over a 5-year period is reported in the light of several reports of complications arising from the application of both clip and spiral electrodes. The performance of 100 electrodes prepared under controlled conditions was analyzed in terms of both the quality of the rate trace produced and of problems experienced during application. Six electrodes were wasted during preliminary handling but of the 94 used in an attempted application to a fetal scalp, 88.3% were considered successful.


Virilization with diffuse involvement of ovarian androgen secreting cells. A case is reported in which virilization of long duration and gradual progression was found in association with ovarian hyperthecosis and bilateral hilar cell lesions. The frequency occurrence of both masculinizing and nonmasculinizing ovarian tumors in association with ovarian hyperthecosis and polycystic ovaries is discussed.


Splenic artery aneurysm rupture in pregnancy. A review and case report. The fourth case of fetal and maternal survival following rupture of a splenic artery aneurysm is presented among 90 reported cases occurring during pregnancy. The previously reported cases are reviewed. A high level of awareness and careful review of history and physical findings in the absence of evidence of the more common causes of concealed hemorrhage may lead to the diagnosis. Ligation of the aneurysm and splenectomy is the indicated therapy.


Lymphangiectasia of the uterus.

Lymphangiectasia of the uterus. A 27-year-old woman, gravida 2, para 2, who presented with a 5-year history of dysfunctional uterine bleeding following a terminated pregnancy is described. Hysterosalpingography suggested a loss of muscle tone and a mass. Pathologic examination revealed a prominent vascular system consistent with diffuse lymphangiectasia.


Necrotizing fasciitis.

Necrotizing fasciitis. Two cases of necrotizing fasciitis, a disease poorly documented in the gynecologic literature because of its extreme rarity, are described. These are the only cases of this entity seen in our department during the last 24 years. Both patients underwent major surgical procedures for uterine stromal sarcoma and mixed mesodermal tumor. In both, the pathognomonic signs, undermining of the skin, and extensive necrosis of the subcutaneous tissue and the underlying fascia, were present. Later gangrene occurred, and both patients died. The literature is briefly reviewed, which emphasizes the importance of immediate surgery when the disease is recognized. The possibility is suggested that the development of such local processes in cases of mixed mesodermal tumor may be an immunologic reaction and the cultured organisms contaminants.


[Quantitative characteristics of the multiplication of muscle cells in the course of postnatal cardiogenesis in mice]. Changes in the absolute number of muscle cells and nuclei in the ventricle during the 1st year of postnatal life were followed in males of the CC57BR mice. The counts were carried out in the cell suspension obtained from the fixed cardiac muscle by means of alkaline dissociation. The number of muscle cells and nuclei reliably increased during the 1st month of life and ceased to increase when the maturity was attained (2-3 months). The number of muscle cells increased twice and that of muscle nuclei almost 4 times. A comparative estimate was made of the role of proliferation and growth of muscle cells in the increase of ventricle weight with the age. The importance of individual variation of the number of muscle cells and nuclei during early postnatal ontogenesis for the study of the myocardium adaptive growth is discussed.


[Incorporation of 3H-thymidine into the cells of the heart conduction system (sinoatrial and atrioventricular nodes, bundle of His) during cardiogenesis in mice]. The number of DNA synthesizing myocytes in the conductive system, as compared with the working myocardium, was determined by means of light autoradiography. At the same time the number of mitosing cells was counted. The indices of labelled nuclei in the sinoatrial and atrioventricular nodes of the embryos after the single and repeated 3H-thymidine injections were significantly lower than in the working ventricle and atrium. During the postnatal development (3-11 days) only in the sinoatrial node the index of labelled nuclei was significantly lower than in the working ventricle. The proliferative activity in all the myocardium parts fell markedly by the 13th day of development. Following 10 3H-thymidine injections with 12 hrs intervals, the indices of labelled nuclei varied from 0.03 to 0.67% in the working ventricle and atrium and from 0.13 to 1.0% in the conductive system of adult mice. During the embryogenesis the index of labelled nuclei in the sinoatrial node was significantly higher than in the atrioventricular one and during the postnatal development-vice versa.


[Experimental-morphological study of morphogenetic potencies of homogeneous aggregates of different types of cells from the freshwater sponge Ephydatia fluviatilis (L.)]. The morphogenetic potencies of somatic cells of the fresh-water sponge Ephydatia fluviatilis in the developing aggregates depend on their initial specialization and the number of cells in the aggregate. The aggregates of nucleolar amoebocytes consisting of 500 or more cells have the highest morphogenetic potencies. All main cell types can arise in the developing homogeneous aggregates of nucleolar amoebocytes. The fine structure of nucleolar amoebocytes at different stages of development of the homogeneous aggregates was studied by means of electron microscopy. The structural rearrangements are described which accompany the process of redifferentiation of the nucleolar amoebocytes in other cell types.


[Formation and reorganization of the early photoreception area in the eyes of sturgeon embryos and prelarvae. Radioautographic study]. A study of the development of eye retina in sturgeons (Acipenser güldenstädti and A. stellatus) by means of autoradiography has shown that during embryogenesis the cells of the high light sensitivity region complete the cycle of proliferation prior to the stage 29, what corresponds to the completion of proliferation of the retina cells in anurans and mammals, rather than at the stage 33, as was considered previously on the basis of the light microscopy morphological studies of differentiation. The cells of definitive retina begin to complete the cycle of proliferation at the stage 35 and the mass completion occurs at the stage 37. Some parameters of mitotic cycle were determined in the cells of definitive retina at the stages 34 and 37 and they proved to be similar with the corresponding parameters of the retina cells in anurans. The time of G2 period equals 2.5 hrs, that of S period 14 hrs and the whole mitotic cycle lasts 21.5 hrs. The transformation of the retina of high light sensitivity region in the definitive retina proceeds, mainly, by means of migration of the cells in this zone from the zone of intensive proliferation surrounding the high light sensitivity region. This transformation proceeds by means of morphological rearrangement of metamorphosis type.


[Effect of cycloheximide on the disintegration of the germinal vesicle membrane and on changes in the cortical layer in oocytes of the clawed toad and the starred sturgeon during maturation on exposure to active cytoplasm]. There was studied the effect of cycloheximide on the changes in the X. laevis and A. stellatus oocytes maturing under the influence of active cytoplasm, taken from the maturing oocytes, and, in the control series, under the influence of progesterone. The suppression of protein synthesis in the X. laevis oocytes by means of cycloheximide by 90--95% did not prevent the amplification of the factor of disintegration of the germinal vesicle membrane after 4 and 5 transfers of the active cytoplasm through the cycloheximide-treated oocytes; no pseudogastrulation was observed in such oocytes. Under the similar suppression of protein synthesis in the A. stellatus oocytes, the germinal vesicle did not break down already after the first injection of active cytoplasm; such oocytes acquried, however, the ability of cortical reaction. The cycloheximide-treated oocytes of both the species did not mature under the effect of progesterone. The fully grown A. stellatus oocytes matured after the injection of active cytoplasm from the mature X. laevis oocytes.


[Effect of continuous illumination on the reproductive function in rats. Aftereffects]. The continuous illumination of 2 months old Wistar rats during 1,2 and 6 months resulted in the appearance of permanent oestrus in 24, 61 and 100% of animals, resp. The weight of newborn rats from the females illuminated during pregnancy was by 7% higher than in the control. The continuous illumination during 1 month prior to mating did not affect the embryogenesis. The weight of 20 days old rats from the females illuminated during 1 month (prior to mating), 2 months (prior to mating + + pregnancy) and 6 months exceeded that of the control ones by 5, 18 and 30%, resp. The increase in the weight of rats persisted in the next reproductive cycle in spite of the standard conditions of light regime (14 hrs day: 10 hrs night).


[Contractility and epithelization in the chick embryo axial mesoderm]. The still unsegemented axial mesoderm was cross cut at different distances form the last somite under the normal physiological conditions, at the low temperature and under the effect of cytochalasin B. Under the normal conditions the inner cells of the axial mesoderm formed a lens-shaped cleft immediately after the operation; this movement was suppressed at the low temperature and absent under the effect of cytochalasin B. This suggests the active contractility of the mesodermal syncytium. The wound margin near the last somite acquired the epithelial structure already within several minutes after the cut, whereas in the more caudal regions it preserved the connective tissue character for a long time. The importance of the axial mesoderm contractility and its ability of epithelization for the somitogenesis is discussed.


[Changes in body weight and respiration rate in loach embryos]. Changes in the raw and dry weight of the intact eggs and isolated embryos of the loach, as well as in the respiration rate have been followed from 3.5 (64 blastomeres) till 46 hrs of development (prior to hatching). The raw weight of the intact eggs was shown to decrease somewhat. The dry weight of the isolated embryos increases twice. The respiration rate of the embryos increases 6.3 times. The intensity of respiration translated per dry and raw weight of the embryos increases as well. The protein content increases more than thrice.


[Effect of lysosome hydrolases on rat embryos]. The application of the lysosome enzymes (acid DNAase, RNAse phosphatase) on the 13th day of pregnancy resulted in the distinct embryolethal effect and the appearance of development abnormalities in the rat embryos. The labilisators of the lysosome membrane weaken the teratogenic activity of the chemical agents whereas the stabilisators strengthen it.


Failure of acupuncture to produce localized analgesia. The pain sensitivity of 49 healthy volunteers has been tested both before and during electro-acupuncture at two or three widely separated places on the body surface. Test areas fell into two categories. The "target" areas were within the part of the body that experienced acupuncturists predicted would be most affected by acupuncture of specified traditional points. "Non-target" control areas fell outside this zone of predicted maximal effect. The subjects and the observers who administered the tests knew that acupuncture anesthesia was being studied, but they did not know which areas were the "targets". Each subject was tested using at least two sensory testing methods. Altogether 4 different tests of pain sensitivity (pinprick discrimination, cold pain ratings, heat pain thresholds and pinch pain thresholds) were used. Small, but statistically highly significant, decreases in pain sensitivity occurred with all 4 tests during acupuncture. However, pain sensitivity fell by the same amount at "target" and "non-target" areas. The variability of the data was sufficiently low that moderate differences between areas would have been detected. The mechanism of "acupuncture anesthesia" is discussed in the light of these negative findings.


A comparison of four tests for assessing the pain sensitivity of different subjects and test areas. Four different testing methods were used to estimate the sensitivity of body surfaces to pain-causing stimuli for a group of 24 healthy male subjects. These methods were (a) determination of the heat pain threshold, (b) determination of the pain threshold for pinching of a skin fold, (c) discrimination of a sharp prick from a blunt contact, and (d) estimation of the pain caused by application of an ice-cold cylinder. Three different body regions were tested: the abdomen, the anterior surface of the neck and the lateral surface of the thigh. Of the three body regions, the thigh area required the strongest stimulation for pain in all 4 tests. The neck was the most sensitive for cold pain, and the abdomen had the lowest threshold for heat pain. There was a barely statistically significant tendency for subjects relatively sensitive on one test to be also relatively sensitive on other tests. Correlation coefficients between any two tests on the same subject were always less than 0.6. Possible reasons for the relative lack of agreement among the results of the different pain tests are discussed.


Experience with early anterior operation in acute injuries of the cervical spine. Of 101 patients with acute injury to the cervical spine below the level of the axis, 74 were selected for anterior operation to ensure stability and comfort and to decompress the neural elements when indicated. Forty-six of the patients underwent a Cloward fusion at the level of the injury. Twenty-eight were treated by bar graft fusion. The indications for these procedures and the techniques involved are outlined. A successful operative result was achieved primarily in 65 of the 74 patients and eventually in 73 patients. The management mortality for patients treated by anterior operation was 2-7 per cent. The advantages of these procedures are the achievement of comfort, the facilitation of early rehabilitation, the maintenance of bony alignment and the preservation and maximal recovery of neurological function.


Cervical spine stenosis.

Cervical spine stenosis. Increased knowledge of the anatomy, physiology and biomechanics of the cervical spine and the enclosed neural and vascular elements is leading to a better understanding of the importance of cervical spine stenosis, as certain clinical syndromes may ensue and may respond well to operative decompression. The present study was designed to investigate the incidence of cervical spine stenosis in patients who have required surgical treatment for cervical spondylosis, and to evaluate the clinical results.


Cervical ankylosis with acute spinal cord injury. Twenty-three cases of acute spinal cord injury in persons with cervical ankylosis are presented. Certain characteristics of major sub-groups are described: ankylosing spondylitis (N = 8), degenerative spondylosis (N = 9) and congenital fusion (congenital non-segmentation) (N = 6). The ankylosing spondylitic group presented a grim prognosis for survival (death rate 50 per cent within 60 days) and for loss of neurological function. Five out of eight cases had permanent neurological loss subsequent to their injuries. Both the ankylosing spondylitic and degenerative spondylotic groups presented problems in diagnosis and medical management. The basic principle is immobilisation of the fracture and mobilisation of the patient. The halo is the technique of choice for fracture immobilisation. An integrated intensive respiratory management programme is essential. Patients with ankylosed spines, particularly those with ankylosing spondylitis, should be educated in simple measures to prevent fracture of their spines.


Somatosensory cortical-evoked responses after feline experimental spinal cord injury. Somatosensory cortical-evoked responses and clinical function were followed in cats for up to 4 weeks after spinal cord compression injury. Despite abolition of cortical responses for from 3 to 19 days after injury, good functional recovery occurred.


Norepinephrine, dopamine and serotonin in experimental spinal cord trauma: current status. The status and function of monoamines in experimental spinal cord trauma remains a subject of controversy and continued debate. The norepinephrine hypothesis has been severely weakened by its lack of reproducibility. Dopamine results are too variable to draw meaningful conclusions. The role of serotonin, particularly with reference to its vasoactive properties and the physiological importance of its effects, remains to be determined.


The effect of medical sympathetic blockage (using reserpine) in the management of spinal cord injury in rats. The effect of medical sympathetic blockage with reserpine was studied in 48 rats following crushing of the cord. Control experiments were carried out with reserpined and unreserpined groups. Marked sensory-motor improvement was found in the reserpined group.


Arm cycloergometry and kinetics of oxygen consumption in paraplegics. A report is given on studies to determine the best methodological approach in exercise testing of paraplegics. Arm cyclometry was found to be very suitable for wheelchair users with normal upper limbs. Oxygen consumption during exercise was studied and analysed by computer.


[Effect of ultrasound on Eimeria tenella (Coccidiida, Eimeriidae)]. Ultrasonics (frequency--20 kHz intensity--11.9 w/cm2 and external lateral surface--1.88 cm2) can annihilate for 8 minutes from 90 to 100% of non-sporulated and for 16 minutes 99.5 to 100% of sporulated oocysts of E. tenella maintained in 10ml suspension plunged into a bath with ice. The destruction rate of oocysts depends on the duration of the ultrasonics effect and the increase of its intensity. It should be noted that the increase in intensity is more effective in this respect than the extension of ultrasonics effect.


[Scanning electron microscopy in studies of parasitic protozoa of fish]. It is expedient to use the scanning electron microscopy for studies of parasitic Protozoa of fishes. The structure of myxosporidians (Henneguya psorospermica, Myxidium rhodei, Chloromyxum truttae) and Chilodenella cyprini was analysed. Microphotos taken by "CWIKscan-100-4" are given. The technique of the preparation of the object for scanning microscopy with regard for its structure is discussed.


[Transovarial and transspermal transmission of Coxiella burneti by the tick Hyalomma asiaticum and its role in the ecology of Q-rickettsiosis]. The transovarial transmission of C. burneti by the tick H. asiaticum was traced on three daughter generations during 751 days. The infection rate of the first generation ticks was found to be 5 times lower than that of the parental ones, of the second generations ticks--9 times lower while F3 population was characterized by 20 fold decrease in the infection rate. The transmission of C. burneti by the infected males of H. asiaticum to the females after copulation was observed in none of the 19 tests. Analysis of the material obtained has shown that the unrestrictedly long "vertical type" maintenance of the Q-rickettsiosis agent in nature is impossible.


[New species of trematodes from fishes of the Falkland-Patagonian region (South-Western Atlantic)]. Four new species of trematodes are described from fishes of the Falkland--Patagonian region as follows: Infundibulostomum patagonicum from Notothenia ramzay; Stenacron mancopsetti from Mancopsetta maculata; Neolepidapedon argentinensis from Salilota australis; Hirudinelloides elongatus from Thyrsites atun. For the latter species a new genus, Hirudinelloides, has been erected. Representatives of the genera Infundibulostomum, Stenacron and Neolepidapedon are first reported from this region.


[Swarming of blood-sucking flies (Simuliidae) in Siberia]. During the swarming period of black flies aggregations of the mass species Schönbaueria pusilla Fries and Simulium truncatum Lundstr. (Jakutia), Byssodon maculata Mg. and S. longipalpe Belt. (Tomsk and Novosibirsk regions, Lower Priamurje) were noted. Observations on the time and character of swarming, statial distribution of swarming black flies and on weather conditions were conducted.


[Method of crossbreeding Trichinella].

[Method of crossbreeding Trichinella]. At the invasional larva stage all the species of Trichinella display different variations in the characters by which the sex can be identified. They should be taken into account in identifying trichinellids by crossing.


[Oxidation of Krebs cycle substrates by Eurytrema pancreaticum mitochondria]. From tissues of E. pancreaticum were isolated mitochondria capable to swell under the effect of some factors. The intensive oxidation of succinate, isocitrate, cisaconitate, oxalacetate and alpha-ketoglutarate by mitochondria and less intensive one of malate, fumarate, citrate and pyruvate were shown. NAD caused the rise in oxidation intensity of isocitrate, cis-aconitate, oxalacetate, alpha-ketoglutarate, malate, fumarate and pyruvate while NADP--of isocitrate and citrate.


[The renin-angiotensin-aldosterone system in hypertensive patients. II. Diagnosis of primary hyperaldosteronism]. Twenty patients suffering from primary hyperaldosteronism were studied. Sixteen had a single adenoma of the adrenal and four had bilateral hyperplasia affecting both adrenals. Cases of primary hyperaldosteronism due to a tumour were characterised by a higher degree of hypermineralocorticism than was seen in the patients with hyperplasia. Plasma aldosterone, after acute volaemic expansion, did not fall below 13 ng/100 ml in the adenoma patients whilst it was lower in the case of hyperplasia. Adrenal phlebography is a useful tool in preoperative diagnosis.


[Morphology of Cucullanus cirratus Müller, 1777 (Nematoda: Cucullanidae)]. The nematodes of C. cirratus of stages III, IV and V were found in the gut of cod Gadus morhua and navaga Eleginus navaga from the White Sea. The morphological data on 176 specimens of C. cirratus were subjected to statistical analysis. The developmental stages can be distinguished by means of measurements of the width and length of pseudo-buccal cavity. The first description of stage IV of C. cirratus and detailed redescription of the IIId stage larvae of C. cirratus are given.


[De novo purine biosynthesis. In vitro measurement in hyperuricemia (author's transl)]. De novo purine biosynthesis has been investigated in circulating blood lymphocytes in vitro. N-formyl-glycinamide ribonucleotide (FGAR) has been mesured using 14C-formate incorporation in the presence of azaserine, a metabolic inhibitor blocking the metabolical pathway at the level of FGAR synthesis. Such a synthesis was measured in 20 healthy controls, 24 patients with primary gout (11 on allopurinol therapy) and 26 patients with chronic renal failure and secondary hyperuricemia (8 on allopurinol therapy). Among gouty patients without allopurinol therapy, FGAR synthesis was normal in 5 and increased in the others. FGAR synthesis was decreased in patients with renal failure whatever the therapy. However, FGAR synthesis remained increased in patients with a primary gout complicated with renal insufficiency. The test we propose for de novo purine biosynthesis measurement is simple and of value to analyse the patho-physiology of hyperuricemia and its therapy. The test allows an acurate discrimination between primary and secondary hyperuricemia in the presence of renal insufficiency.


[Increased hematological toxicity of antineoplastic drugs with simultaneous androgenotherapy (author's transl)]. Thirty two patients with malignant lymphoma - mainly Hodgkin's disease - were randomized for simultaneous treatment by high doses of metenolone during MOPP chemotherapy, to reduce its hematological toxicity. The results have shown surprisingly an increased hemato-toxicity in patients receiving androgens, with significantly more marked anemia and thrombocytopenia, reducing the total doses of anti-cancer drugs. This side effect could be explained by a cycling of the hematopoietic stem-cells and call to some caution when androgens are used during cancer chemotherapy.


[The nephrotoxic potential of cefalotin-aminoside antibiotic combinations (author's transl)]. The combined study of animal experimental findings and clinical trials indicates the minimal effect on renal function of a combination of Cefalotin with aminoglycosides. Nevertheless, whilst the published cases of renal insufficiency are often difficult to analyse with regard to the exclusive responsibility of antibiotic therapy, it seems that strict attention to dose and frequency of administration is essential when this combination is prescribed and that regular surveillance of renal function is necessary, above all with prolonged treatment. The renal toxicity of aminoglycosides more recent than Gentamicin has not yet been fully explored, the same applying to their use in association with Cefalotin.


[Double contrast hypotonic duodenography]. Double contrast hypotonic duodenography, as carried out by the authors, is a simple and rapid examination, suitable for routine use. At the same time, it includes a double contrast study of the oesophagus and stomach. The normal appearance is easily analysed, in particular that of the great caronculus. It is a selective examination of the duodenal outline, playing an essential role in the detection of lesions of the latter as well as in adjacent organs which may alter its shape. It provides only a suggestion as to aetiology and must therefore be completed, when appropriate, by other studies.


Membrane associated cytoplasmic mRNA in Artemia salina; functional and physical changes during development. The physical and functional properties of the mRNA population from developing embryos of the brine shrimp Artemia salina were characterized. About 20% of the total poly(A)-rich mRNA in these embryos appears to be specifically associated with the membrane  fraction throughout early development, and physically differs markedly from the free cytoplasmic mRNA. The membrane-associated mRNA fraction consists of two well-defined populations of molecular weight of 5.2x10(5) and 3.6x10(5), whose relative amount changes during the various stages of embryo development. The size of the poly(A) tail at the 3'-end of the mRNA molecules, as estimated by processive phosphorolysis, was found to consist of 180 and 210 adenosine residues for the two respective mRNA species. The in vitro translation products of the membrane-bound mRNA molecules are apparently similar to those of the free mRNA molecules.


DNA sequence at the end of the cI gene in bacteriophage lambda. The nucleotide sequence of 57 base pairs near the end of the cI gene in phage lambda is presented. This sequence was determined by direct sequencing techniques and includes the codons for 11 carboxyterminal aminoacids of the cI product, the lambda repressor. The sequence reveals that the cI gene, which has recently been shown to have a unique initiation region, is terminated by a UGA codon. A GUG triplet, which could act as a translation start signal for the rex gene occurs 8 base pairs beyond the cI termination codon. This GUG triplet is preceded by a sequence that could serve as a strong ribosome binding site for the rex message.


Digestion of insect chromatin with micrococcal nuclease, DNase I and DNase I combined with single-strand specific nuclease S1. The chromatin of the lepidopteran Ephestia kuehniella was digested by micrococcal nuclease, DNase I and S1-nuclease combined with DNase I pretreatment. The resulting DNA fragments were analyzed by gel electrophoresis and compared with the DNA fragments of rat liver nuclei obtained by the same process. Extensive homology was revealed between insect and mammalian chromatin structure. The combined DNase I- S1-nuclease digestion yields double-stranded DNA fragments of lengths from 30 to 110 base-pairs. These DNA fragments are not obtained from nuclei predigested extensively with micrococcal nuclease. The results are discussed with respect to the internal structure of the chromatin subunit.


A kinetic study of 1H leads to 3H exchange in C(8) H-groups of purinic residues in DNA. The kinetic study of 1H leads to 3H exchange in C(8) H-groups of purinic residues of DNAs with different G-C content as well as in corresponding dNMP mixtures have been carried out. The present results show that 1H--3H exchange in DNA is retarded (as compared to the exchange in dNMP mixtures) to a lesser extent (Kret =2.4-2.8) than in RNA (Kret=6-8). The degree of retardation in these polymers is practically independent of their nucleotide composition. Assuming the ylide mechanism of exchange reaction it is suggested that the lower rate of 1H leads to 3H exchange in C(8) H-groups of purinic residues in polynucleotides of A-form (RNA and other polyribonucleotides) as compared to those of B-form (DNA and other polydeoxyribonucleotides) might be accounted for by decreased availability of C(8) H-groups for OH-ions of the solvent due to a different microenviroment of these groups in A- and B-type helixes.


DNA regions essential for the function of a bacteriophage fd promoter. The promoter for the major coat protein gene of bacteriophage fd contains a unique sequence. TATAAT, in the non-transcribed region corresponding to the Pribnow box. A R-Hha I cleavage site which destroys functions is located five pairs upstream from the TATAAT sequence (fifteen base pairs upstream from the RNA initiation site). The promoter was cleaved into two fragments by R-Hha I and each promoter fragment was joined to DNA fragments derived from other regions. Ligation of the TATAAT-containing fragment to any of the DNA fragments examined resulted in recovery of promoter function. The results suggest for this type of promoter that no unique sequence is necessary upstream from the R-Hha I cleavage site although a contiguous DNA chain must be present in this area.


Synthesis of acylamino acid esters of nucleoside 5'-phosphates and their investigation with PMR and CD spectra. The acylamino acid esters of nucleoside 5'-phosphates are synthesized via condensation of N-(N'-acylaminoacyl) imidazoles with nucleoside 5'-phosphates. The PMR and CD spectra of the esters obtained are studied. The 3'-isomers of the substances under study are observed to have a shift in the conformational N in equilibrium S equilibrium of the carbohydrate moiety in favour of the S-form as compared to the initial nucleosides and their 2'-acyl esters.


The nucleotide sequence of two restriction fragments located in the gene AB region of bacteriophage S13. The nucleotide sequence of a double stranded DNA fragment from the gene AB region of bacteriophage S13 DNA has been determined. The fragment was isolated as two adjacent shorter fragments by cleavage of S13 replicative form (RF) DNA with restriction endonuclease III from Hemophilus aegyptius. The strands of the fragments were separated electrophoretically and hydrolyzed with T4 endonuclease IV to yield short oligonucleotides which were then sequenced by partial exonuclease digestion. The complete nucleotide sequence of the restriction fragments was obtained by ordering the inter- and intrastrand overlapping oligonucleotide sequences. The adjacent fragments were 190 nucleotides in length. The sequences included a HindII site, an AluI site and two sequences which may be possible transcription initiation sequences, one with an adjacent sequence homologous to the canonical promoter site sequence T-A-T-Pu-A-T-Pu. Examination of the three possible reading frames for translation of the sequence revealed only one possible complete translation product. The postulated partial sequence of gene A protein has a highly positively charged arginine-rich area which may have importance in DNA binding.


Mechanism of tRNA-synthetase recognition: role of terminal A. The function of the terminal A of tRNA Phe (yeast) with respect to complex formation with the cognate aminoacyl-tRNA synthetase has been studied using equilibrium and fast kinetic techniques. Removal of the terminal A influences the equilibrium parameters of the tRNA-synthetase interaction only slightly, the mechanism of complex formation, however, is changed significantly. The binding mechanism of unmodified tRNAPhe comprises a recombination step and a consecutive conformational change. In contrast, the reaction between tRNAPheCC and the cognate synthetase is characterized by a simple one step mechanism. It is concluded that the terminal A is responsible for the occurrence of the conformational change of the tRNA-synthetase complex. The conformational change is interpreted as a proper alignment of the terminal A of the tRNA to the active site of the synthetase.


Protection of particular endonuclease R. Hind III cleavage sites by distamycin A, propyl-distamycin and netropsin. It is shown that three related antibiotics, distamycin A, propyl-distamycin and netropsin, can protect certain endo R.Hind III cleavage sites from attack by endonuclease, giving rise, after endo R.Hind III digestion, to larger DNA fragments. Bacteriophage lambda DNA has six recognition sites for Hind III enzyme. Three of these sites: shind III 2, 3 and 6 can be protected from nuclease action by all the antibiotics used. Propyl-distamycin protects partly shind III 5, too. Netropsin protects partly sites shind III 5 and 4, while distamycin A protects all the sites but shind III 1 so the Hind III digestion produces only two large fragments of lambda DNA.


Chromatin very small angle neutron scattering: further evidence for a 30 nm diameter super coil in dilute solutions. Intact chromatin, chromatin minus histone H1, and nuclease digestion fragments have been studied by very small angle neutron scattering. The results are not consistent with a straight chain of nucleosomes and require the presence of a higher order coiling in monovalent salt solutions. The data are interpretable by a structure having a cross section radius of gryration of 8.5 +/- 1 nm, which suggests an outer diameter for a coil of nucleosomes of 27 +/- 3 nm.


Effect of non-histone proteins on thermal transition of chromatin and of DNA. The effect of chromatin non-histone protein on DNA and chromatin stability is investigated by differential thermal denaturation method. 1) Chromatin (rat liver) yields a multiphasic melting profile. The major part of the melting curve of this chromatin is situated at temperatures higher than pure DNA, with a distinct contribution due to nucleosomes melting. A minor part melts at temperatures lower than DNA which may be assigned to chromatin non-histone protein-DNA complex which destabilized DNA structure. 2) Heparin which extracts histones lowers the melting profile of chromatin and one observes also a contribution with a Tm lower that of pure DNA. In contrast, extraction on non-histone proteins by urea supresses the low Tm peak. 3) Reconstitution of chromatin non-histone protein-DNA complexes confirms the existence of a fraction of chromatin non-histone protein which lowers the melting temperature when compared to pure DNA. It is concluded that chromatin non-histone proteins contain different fractions of proteins which are causing stabilizing and destabilizing effect on DNA structure.


Specific hydrolysis of rabbit globin messenger RNA by S1 nuclease. S1 nuclease isolated from Aspergillus oryzae has been used to investigate the secondary structure of rabbit globin messenger RNA (mRNA). The enzyme, which is specific for single stranded nucleotides, digests globin mRNA to a limited extent, with 65-75% of the mRNA nucleotides resistant to digestion under mild conditions. This limited digestion is not due to enzyme inactivation, but rather to the normal activity of the single-strand nuclease. The reaction was studied as a function of temperature, salt and enzyme concentration. Analysis of the products of digestion on 20% acrylamide- 7M urea slab gels reveals a stable pattern of unique fragments ranging in size from 9 to 71 nucleotides. Separated alpha and beta globin mRNAs show similar, but not identical gel patterns, indicating strong structural similarities between the two species. The high degree of nuclease resistance, along with the fragment patterns seen on polyacrylamide gels, gives evidence to support a model of rabbit globin mRNA which contain specific, rather than random, helical structure.


Nucleoside 3'-phosphotriesters as key intermediates for the oligoribonucleotide synthesis. IV. New method for removal of 2,2,2-trichloroethyl group and 31P NMR as a new tool for analysis of deblocking of internucleotide phosphate protecting groups. Zinc/acetylacetone/pyridine treatment has been designed as a very efficient method for removal of 2,2,2,-trichloroethyl group from phosphoesters. Internucleotide and terminal 2,2,2-trichloroethylphosphotriesters were transformed to corresponding diesters quantitatively. Much less reactive 2,2,2-trichloroethylphosphodiesters produced monoesters with ca. 90% yield. 31P NMR spectroscopy has been proposed as a new tool for analysis of removal of internucleotide phosphate protecting groups-a crucial step in oligonucleotides synthesis via phosphotriester approach.


Nucleotide sequence of 3' untranslated portion of human alpha globin mRNA. We have determined the nucleotide sequence of 75 nucleotides of the 3'-untranslated portion of normal human alpha globin mRNA which corresponds to the elongated amino acid sequence of the chain termination mutant Hb Constant Spring. This was accomplished by sequence analysis of cDNA fragments obtained by restriction endonuclease or T4 endonuclease IV cleavage of human globin cDNA synthesized from globin mRNA by use of viral reverse transcriptase. Analysis of cRNA synthesized from cDNA by use of RNA polymerase provided additional confirmatory sequence information. Possible polymorphism has been identified at one site of the sequence. Our sequence overlaps with, and extends the sequence of 43 nucleotides determined by Proudfood and coworkers for the very 3'-terminal portion of human alpha globin mRNA. The complete 3'-untranslated sequence of human alpha globin mRNA (112 nucleotides including termination codon) shows little homology to that of the human or rabbit beta globin mRNAs except for the presence of the hexanucleotide sequence AAUAAA which is found in most eukaryotic mRNAs near the 3'-terminal poly (A).


Raman spectral studies of nucleic acids. XVII. Conformational structures of polyinosinic acid. Laser-Raman spectra of poly(rI) show the formation of an ordered complex in aqueous solutions of high ionic strength. This structure exhibits the A-helix geometry, contains stacked bases and is apparently stabilized by specific hydrogen bonding involving hypoxanthine C6=0 groups. Thermal dissociation of the poly(rI) complex (Tm=45 degrees C) yields single-stranded and disordered poly (RI) chains. A disordered structure also occurs for poly (rI) in aqueous solutions of low ionic strength. In oriented films, poly (rI) forms an ordered structure probably the same as that which occurs in solutions of high ionic strength. Raman intensities measured at 815 and 1100 cm-1 in spectra of poly (rI) and poly (rU)-poly (rA)-poly(rU) indicate that the correlation previously established for single- and double-stranded ribopolymer structures is valid also for these multi-stranded structures. X-ray diffraction and model-building studies confirm the A-helix structure.


Photoproducts from DNA pyrimidine bases and polycyclic aromatic hydrocarbons. The major photoproduct formed between benzo[a]pyrene and thymine is identified as 1-(benzo[a]pyren-6-yl)-thymine by means of spectroscopic analysis and isotopic syntheses. Irradiation of 1-methylcytosine hydrochloride and anthracene gives two isolable photoproducts of which one is assigned the structure 5-(anthracen-9-yl)-1-methylcytosine.


A simple, rapid preparation of alpha[32P]-labelled adenosine diphosphate. Hexokinase (EC 2.7.1.1) will convert commercially available alpha-[(32)P]-labelled ATP into alpha-[(32)P]-labelled ADP. A simple, rapid isolation procedure for the alpha-[(32)P]-labelled ADP is described and this synthetic method can be used for the preparation of other alpha-[(32)P]-labelled nucleoside diphosphates.


A simple, general procedure for purifying restriction endonucleases. A simple, general method for purifying restriction endonucleases is described. The method employs precipitation of nucleic acids from crude extracts with polyethyleneimine followed by affinity chromatography on columns of heparin covalently linked to agarose. Most of the sixteen enzymes tested could be purified to a degree sufficient for DNA sequencing work by this method sometimes supplemented by at most one step of ion exchange chromatography.


Structure of poly 8-bromoadenylic acid; conformational studies by CPF energy calculations. Poly 8-bromoadenylic acid [poly(BBrA)] is the only known all-syn polynucleotide. It shows a helix-coil transition with a melting curve centred around 55 degrees C. Energy calculations based on classical potential functions have been used to explore the three-dimensional structure of this polymer in helix and random coil. It is concluded that the ordered state is a helix of two parallel strands with a two-fold rotation axis, and the duplex is stabilised by hydrogen bonds involving N1 and H6. Each strand has a conformation with C3' endo geometry, phi' = 216 degrees, omega' = 280 degrees, omega = 294 degrees, phi = 179 degrees, chi = 243 degrees and psi = 57 degrees. Such a conformation leads to approximately 8 nucleotide units per turn of the helix and an axial rise of 3.9A degrees. The structure of poly(8BrA) has been compared with that of the related polymer poly(A) which forms a double helical structure in acidic conditions with bases in the anti conformation and with interstrand hydrogen-bonds between N7 and H6. This is the first time that a specific geometrical model of a novel polynucleotide structure has been produced initially by potential energy calculations, though such calculations on a number of known structures have been reported previously.


Chemical synthesis of the hexanucleotide d(A-C-C-A-G-C) required to isolate fibroin mRNA on an affinity column. The synthesis of the hexanucleotide d)A-C-C-A-G-C), complementary to the 2 major triplets of fibroin mRNA, using the phosphotriester methodology is described. The protected dinucleotides ((MeO)2Tr)dbzA.anC, ((MeO)2Tr)danC.bzA and ((meO)2Tr)dacG.anC were synthesized; the latter two were detritylated and joined in stepwize fashion to the 1st to form the protected hexanucleotide ((MeO)2Tr)dbzA.anC.anC.bzA.acG.anC. The latter was deblocked with NH3 and acid to form the hexanucleotide d(A-C-C-A-G-C). In view of the ability of a prototype affinity column, oligo dC-cellulose, to isolate fibroin mRNA, prospects appear excellent for the d(A-C-C-A-G-C)-cellulose affinity column isolation of fibroin mRNA.


Nuclear magnetic resonance of nucleotide-amidates. Investigation of the secondary structure of methylester of deoxyadenylyl-(5'-3')-deoxyadenylyl-(5' leads to N)-L-phenylalanine. The secondary structure of methyl ester of deoxyadenylyl- - (5' - 3') - deoxyadenylyl - (5' leads to N) - L-phenylalanine / L- PheOMe - d(pApA)/ was investigated using proton magnetic resonance. The folded conformation stabilized by hydrophobic interactions in aqueous solution was shown for this compound. The investigation of the chemical shifts of aromatic protons of L- PheoOMe - d(pApA) permitted us to construct the conformational model of this compound.


Polynucleotides. XLV Synthesis and properties of poly(2'-azido-2'-deoxyinosinic acid). Poly (2'-azido-2'-deoxyinosinic acid), [poly (Iz)], was synthesized from 2'-azido-2'-deoxyinosine diphosphate by the action of polynucleotide phosphorylase. Poly (Iz) has UV absorption properties similar to poly (I) and hypochromicity of 11% at 0.15M Na+ and neutrality. In solutions of high Na+ ion concentration, poly (Iz) forms a multi-stranded complex and its Tm at 1.0M Na+ ion concentration was 43 degrees. Upon mixing with poly (C), poly (Iz) forms a 1:1 complex having a Tm lower than that of poly (I)-poly (C) complex in the same conditions. The effect of substitution at the 2'-position of the poly (I) strand was discussed in relation to the interferon-inducing activity.


Endogenous DNA polymerase activity in fractionated rat lever chromatin. Chromatin isolated from adult rat liver was fractionated into template active and inactive components by controlled shearing and glycerol gradient centrifugation. The fractionated chromatin was assayed for DNA-dependent DNA polymerase (DNA mucleotidyl transferase EC 2.7.7.7) activity with and without exogenous activated DNA serving as template. With endogenous chromatin as template, it was found that 90% of the endogenous chromatin bound DNA polymerase activity was located in the transcriptionally active fraction of chromatin, while the distribution of DNA polymerase assayed with exogenous activated DNA was found to be 65% in the transcriptionally active and 35% in the inactive fractions. However, when DNA polymerase was solubilized from these fractions by salt extraction, enzyme activity was found to be equally distributed, suggesting that the difference in endogenous DNA polymerase activity observed between eu- and heterochromatin is due to the restricted template found in repressed fractions.