Effects of methapyrilene on DNA synthesis in mice and rats following continuous dietary exposure.

The effects of the antihistamine methapyrilene (MP) on DNA synthesis in rats and mice were investigated. Previous studies have demonstrated a dose response for tumor induction in the rat but no carcinogenic effect in the mouse. To study the role of DNA synthesis in MP carcinogenesis, rats and mice were administered MP at doses of 0, 62.5, 125, 250 or 1000 p.p.m. in the diet for a period of 1-12 weeks. Bromodeoxyuridine was administered continuously using an osmotic minipump during the last week of treatment to provide an index of DNA synthesis. Results demonstrated that in the rat 250 and 1000 p.p.m. MP increased DNA synthesis in a dose-dependent manner that correlated with the tumor response in previous oncogenic studies. MP at 62.5 p.p.m. did not increase DNA synthesis, indicating a no effect level for cell proliferation and suggesting a no effect level for carcinogenicity by this compound in the rat. MP did not induce DNA synthesis in mice after exposure to 1000 p.p.m. for 12 weeks, nor did it induce changes in serum chemistries or liver histopathology suggestive of overt toxicity as was seen in the rat at 1000 p.p.m. The correlations between labeling index and tumorigenicity in the rat and mouse strongly support a role of cell proliferation in the carcinogenic mechanism of MP.


Rat hepatocyte nodules are resistant to the necrogenic effect of D-galactosamine.

D-Galactosamine is a known hepatotoxin which induces liver cell necrosis via depletion of UTP and other uridine nucleotides. Our previous work indicated that nodular hepatocytes have higher levels of total uridine nucleotides compared to normal liver, and in the present study we investigate the effect of galactosamine treatment on hepatocyte nodules and surrounding liver. Hepatic nodules were generated in male Wistar rats according to the Solt and Farber protocol. Six months after initiation animals received a single injection of D-galactosamine (500 mg/kg i.p.) and were then killed 1, 2, 4 or 7 days later. Histological analysis of liver revealed the presence of extensive liver cell necrosis in normal tissue 1 and 2 days after galactosamine treatment. However, very little or no necrosis was detectable inside hepatic nodules at any time point, indicating that these focal areas are resistant to the cytotoxic effect of galactosamine. This type of resistance could be the expression of a new component in the resistant phenotype of hepatic nodules.


Some quantitative considerations about DNA adduct enrichment procedures for 32P-postlabelling.

The concentrations of 2'-deoxyribonucleoside-3'-monophosphates remaining in calf-thymus DNA digests after nuclease P1 digestion or extraction into 1-butanol, the most commonly used adduct enrichment procedures prior to the application of the 32P-postlabelling assay, were measured using HPLC and 32P-postlabelling methods. When 10 micrograms of DNA digested to mononucleotides was used, the total amount of nucleotides remaining in the samples were approximately 4 and approximately 14 pmol after nuclease P1 treatment or 1-butanol extraction respectively. The influence of various concentrations of normal nucleotides on the labelling efficiency of a 2'-deoxyguanosine-3'-monophosphate adduct of benzo[a]pyrene diol-epoxide was also studied and found to depend upon the ratio of normal nucleotides/adducted nucleotides present in the sample. Also, the ATP/normal nucleotides ratio in the phosphorylation reaction may affect the quantitation of the adducts and thus deserves due consideration.


Modulation of N-nitrosomethylbenzylamine bioactivation by diallyl sulfide in vivo.

Diallyl sulfide (DAS), a major component of garlic oil, is an inhibitor of tumorigenesis by various metabolically activated carcinogens. In rats, pretreatment with DAS has been observed to suppress completely the induction of oesophageal neoplasms by N-nitrosomethylbenzylamine (NMBzA) (Wargovich et al. (1988) Cancer Res., 48, 6872-6875). This communication reports the effects of DAS on overall NMBzA metabolism and on DNA methylation of NMBzA in vivo under conditions equivalent to a single treatment of the chemoprevention assay. Male Fischer 344 rats received a single i.g. dose of DAS (200 mg/kg body wt) followed by an s.c. injection of [methyl-14C]NMBzA (3.5 mg/kg). In controls, exhalation of 14CO2 was complete within 5 h (t1/2max = 1.2 h), with 50% of the injected radioactivity recovered as 14CO2. When DAS was given 3 h prior to [methyl-14C]NMBzA, 49% of the injected radioactivity was released within 10 h (t1/2max = 3 h). When DAS was administered 18 h before the carcinogen, 42% of [methyl-14C]NMBzA was converted to 14CO2, with exhalation complete after 6 h (t1/2max = 1.8 h). We further examined the effects of acute doses of 10-200 mg/kg of DAS on DNA methylation by a single dose of NMBzA (3.5 mg/kg; survival time, 6 h) administered 3 h later. At 200 mg/kg, DAS inhibited the formation of O6-methyldeoxyguanosine (O6-MEdG) in oesophagus (-26%), nasal mucosa (-51%), trachea (-68%) and lung (-78%). In liver, levels of 7-MEdG were reduced by 43%. Decreases in DNA methylation were proportional to dose for > 25 mg/kg of DAS in oesophagus, liver and nasal mucosa, for 25-200 mg/kg in trachea and 10-50 mg/kg in lung. The dose-activity relationship for inhibition by DAS of DNA methylation by NMBzA suggests that short-term modulation of carcinogen bioactivation in situ contributes to but may not be sufficient for the chemo-prevention of nitrosamine tumorigenesis by DAS.


Alterations in nucleotide pools in rats fed diets deficient in choline, methionine and/or folic acid.

The fidelity of DNA synthesis is critically dependent on the correct balance and availability of the deoxynucleoside triphosphate (dNTP) precursors for the polymerases involved in replication and repair. Since folate-derived one-carbon groups are essential for the de novo synthesis of both purines and pyrimidines, the purpose of the present investigation was to determine whether diet-induced depletion of folates would alter intracellular dNTP pools. Fischer 344 rats were fed one of four semi-purified diets for a period of 8 weeks: (i) supplemented control; (ii) deficient in folic acid; (iii) deficient in methionine and choline; and (iv) deficient in methionine, choline and folic acid. In contrast to natural diets, semi-purified diets are nucleotide-free and consequently lack substrates for salvage pathway synthesis. This omission may place unusual stress on folate-dependent de novo nucleotide synthesis especially under conditions of dietary methyl-donor deficiency. Reversed-phase HPLC analysis of dNTP in spleen cell extracts indicated that both the thymidylate monophosphate and thymidylate triphosphate pools were decreased in spleen cells from the deficient rats consistent with a decrease in folate-dependent de novo synthesis. In addition, purine biosynthesis appeared to be negatively affect by methyl-donor deficiency as evidenced by a reduction in dGTP and dATP pools. These data indicate that deoxynucleotide pool imbalance, well known to produce cytogenetic and mutagenic events in vitro, can also be induced in this in vivo model of diet-induced carcinogenesis.


Effects of hyperthermia on intracellular calcium concentration and responses of cancerous mammary cells in culture.

Effects of hyperthermia on the intracellular calcium concentration (Cai) of an established mouse breast cancer cell line, MMT060562, were studied using fura-2 fluorescence microscopy and the whole-cell clamp technique. A sudden change of temperature from 37 to 45 degrees C induced a transient increase in the fluorescence ratio permeability of the cell membrane and inward current. Deletion of extracellular calcium abolished the fluorescence ratio response to the rise in temperature. Cai of some cells increased after hyperthermia treatment at 44-48 degrees C for 20 min, but the average increase of Cai was negligible. After hyperthermia treatment, spontaneous oscillation of Cai, chemical responses to ATP and bradykinin and the mechanically-induced spreading response diminished. However, the mechanically induced increase of Cai within the stimulated cell remained even after hyperthermia treatment. Suppression of the ATP-induced Cai response recovered to about half the original level within 12 h. Blockage of protein synthesis with cycloheximide (100 microM) had no effect on the recovery. The D-myo-inositol 1,4,5-triphosphate (IP3)-dependent increase of Cai remained intact even after hyperthermia treatment. It is concluded that hyperthermia treatment increases both the permeability of the cell membrane and Cai, but decreases the sensitivity of cells to ATP and bradykinin, presumably due to modification of the signal transduction mechanism.


Studies on the ATPase of Bacillus cereus.

The membrane ATPase (EC 3.6.1.3) of Bacillus cereus was solubilized by a 'shock-wash' process and purified. The non-specific phosphatase contaminant was separated by glycerol density gradient centrifugation. The optimum temperature was 39.5 degrees C and the pH optimum at 7.5. On SDS-polyacrylamide gel electrophoresis two classes of subunits were observed in equal proportions with molecular weights of 70 K and 83 K. The effect of various compounds on the enzymatic activity was studied. The enzyme was insensitive to NaN3, oligomycin and to divalent cations, but was inhibited by citrate and oxalate.


Ornithine decarboxylase and ornithine decarboxylase-inhibiting activity in rat thymocytes.

Isolation of thymocytes from rat thymus resulted in the disappearance of the high activity of ornithine decarboxylase (ODC) that characterizes the thymus of young rats, together with the appearance of an antizyme-like ODC inhibiting activity, which showed a chromatographic profile that resembled that of dexamethasone-treated rat thymus. Omission of serum or addition of dexamethasone or spermidine did not affect appreciably the extent of the antizyme-like activity. On the other hand, a variety of hormonal effectors, i.e. insulin, glucagon, adrenalin and T3, as well as the phorbol ester, PMA or the mitogen, concanavalin A (Con A) induced ODC activity in cultured thymocytes together with the disappearance of the antizyme-like activity. A paradoxical, transient induction of ODC was caused by the transcriptional inhibitor, actinomycin D. Complexed ODC was detected in rat thymus, but not in thymocytes, either quiescent or stimulated by mitogens. These results indicate that thymic lymphocytes can express either ODC activity or its inhibitor depending on the hormonal and proliferative status of the cells.


Polymerase chain reaction in the detection of mRNA transcripts from the slow skeletal troponin T (TNNT1) gene in myotonic dystrophy and normal muscle.

Recent studies have shown that the gene encoding for the slow skeletal troponin isoform T (TNNT1) is located on the proximal long arm of human chromosome 19 in the myotonic dystrophy (DM) region. In order to test TNNT1 as a candidate gene for DM, we have isolated TNNT1 cDNA from skeletal muscle from two healthy individuals and from two patients with DM. Sequencing of the TNNT1 cDNA from the DM and normal muscle revealed two sequence variants but no transcriptionally significant mutations. This work rules out a defect in the coding segment of TNNT1 as a cause of DM and provides a polymerase chain reaction protocol for studying troponin T gene expression.


Phosphate efflux from jejunal enterocytes of the rat: effect of phosphate concentration gradient and pH.

The effect of variation in the intracellular and extracellular phosphate concentration on the Pi efflux across the basolateral membrane of pre-loaded enterocytes has been examined. Efflux rate constants for Pi fell from 0.89 h-1 to 0.68 h-1 as the extracellular Pi concentration was increased from 0.5 mM to 5 mM. As the intracellular Pi concentration was raised from 0.5 to 3 mM the rate constant dropped from 0.95 h-1 to 0.77 h-1. The findings are indicative of the presence of Pi-specific transporter at the basolateral membrane. The efflux rate constant of Pi at pH 7.1 was higher than that at pH 7.4 suggesting that the Pi flux across the basolateral membrane of enterocytes follows a similar pattern towards pH changes as do fluxes across the brush-border membrane.


Effect of streptozotocin-induced diabetes on the handling of phosphate by the rat small intestine.

The effect of chemically-induced diabetes on the handling of phosphate (Pi) by rat jejunal enterocytes has been investigated in the presence of a Na- or a choline-gradient. Pi uptake was significantly increased in both gradients. The Pi efflux rate constants for enterocytes from diabetic rats were similar to those of control rats. The effect of diabetes on both the protein and alkaline phosphatase isoenzymes of the rat small intestinal brush-border membranes was examined using SDS-PAGE. The patterns given by membranes from rats 14 days after the induction of diabetes were no different from those of controls.


Effect of inhibitors and activators of tyrosine kinase on insulin imprinting in Tetrahymena.

Primary exposure of Tetrahymena cells to insulin gave rise to hormonal (insulin) imprinting in the offspring generations, as judged from the increase in binding upon reexposure to insulin. Vanadate mimicked the action of insulin, inasmuch as it also induced imprinting for insulin, whereas the other tyrosine kinase activator tested, namely H2O2, had no such effect. However, combined treatment with vanadate+H2O2 + insulin induced a more pronounced imprinting for insulin than either insulin or vanadate on their own. The tyrosine kinase inhibitor genistein, a plant flavonoid, did not change the value for insulin binding significantly relative to the control immediately after exposure, but increased it slightly in the offspring generations after 24 h at high dilution. Upon combination with insulin, 10(-4)M genistein inhibited imprinting by insulin. These experimental observations suggest that there may be a key role for tyrosine kinase activity in the mechanism (development) of imprinting.


In situ localization of male germ cell-associated kinase (mak) mRNA in adult mouse testis: specific expression in germ cells at stages around meiotic cell division.

Biochemical analysis of the male germ cell-associated kinase (mak) gene, which was isolated recently by using weak cross-hybridization with the v-ros tyrosine kinase gene, revealed that the gene was highly expressed in mammalian testicular germ cells, but not in ovarian cells. In order to identify the cells which express the mak gene in more detail, we localized mak mRNA in frozen sections of mouse testis by non-radioactive in situ hybridization. In this study, we utilized thymine-thymine (T-T) dimerized mak cDNA as a haptenic, non-radioactive probe, and the signal was detected enzyme-immunohistochemically by using an anti-T-T antibody. As a result, mak mRNA was localized intensely in late pachytene (stage X) and diplotene (stage XI) spermatocytes, and faintly in dividing spermatocytes (stage XII) and early round spermatids (stage I-II), suggesting that the gene may play an important role in the phase around meiotic cell division, but not throughout the entire meiosis.


Normal and Ha-ras-1 oncogene transformed Buffalo rat liver (BRL) cells show differential resistance to cytoskeletal protein inhibitors.

In the present study, using immunofluorescence microscopy, we have demonstrated that normal and Ha-ras-1 transformed Buffalo rat liver (BRL) cells which were exposed to cytoskeletal protein inhibitors, showed a differential resistance of their microfilament and microtubule networks. One hour exposure of normal BRL cells to 10(-5) M cytochalasin B provoked a clear and already total breakdown of actin filaments. However, at this concentration of cytochalasin B, the microfilaments of transformed BRLHO6T1-1 cells were not seriously affected; a higher cytochalasin B concentration (> or = 2 x 10(-5) M) was required to induce a significant breakdown of microfilaments in these transformed cells. The two cell lines also demonstrated differential microtubule stability when they were treated with either colchicine or triethyllead. Three hours exposure to 10(-6) M of either antimicrotubule agents was sufficient to disrupt the microtubules of normal BRL cells, without affecting their counterparts in the transformed BRLHO6T1-1 cells. A 10-fold higher drug concentration (10(-5) M) was required to induce microtubular breakdown in the transformed BRL cells. The differential stability of microfilaments and microtubules in normal and transformed BRL cells that was observed could not be attributed to a differential internalization of the agents, as shown by experiments on the uptake of [3H]-cytochalasin B and triethyllead. In addition, the transformed BRLHO6T1-1 cells did not express altered actin and tubulin isoforms, as demonstrated by isoelectric focusing followed by immunoblotting analysis. We conclude that the transformation of BRL cells with the Ha-ras-1 oncogene results in a greater stability of microfilaments and microtubules, leading to a structurally firmer cell shape.(ABSTRACT TRUNCATED AT 250 WORDS)


Sexual and ontogenic differences in K(+)-induced gonadotropin and prolactin release in vitro.

Ontogenic and sexual differences have been described in the regulation of anterior pituitary hormone release. In the present experiments we studied basal release and the effect of a depolarizing concentration of K+ on in vitro gonadotropin and prolactin release from anterior pituitaries of male and female rats at 12, 20 and 28 days of age. Basal release of LH and FSH increased with age, values obtained from female glands being significantly higher than those obtained from male glands. K(+)-induced release of LH did not present differences among ages, although the response in females was always greater than that in age-matched males. If K(+)-induced release of LH was considered in relation to basal release, infantile 12-day-old rats of both sexes, had a significantly greater sensitivity to the effect of K+ in comparison to older ages, as has been described for the LH-releasing effect of LHRH and of other stimuli. K(+)-induced FSH release was maximal in females at 20 days of age, and in males at 28 days of age. Percentage increase relative to basal values, induced by K+ was also greatest at 12 days in both sexes, although values from female glands were significantly higher than those from males. Basal and K(+)-induced prolactin release increased significantly with age in both sexes. Basal prolactin release was greater in females than in males at 28 days of age, and no other sexual difference was evidenced.(ABSTRACT TRUNCATED AT 250 WORDS)


Expression of neurotrophins and the low-affinity NGF receptor in septal and hippocampal reaggregate cultures: local physiologic effects of NGF synthesized in the septal region.

Nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) are members of a family of trophic factors designated the neurotrophins, each of which can bind to the low-affinity NGF receptor (LNGFR). To investigate the mechanisms that regulate the expression of the neurotrophins and the LNGFR in the developing brain, we grew cells from the embryonic mouse septum and hippocampus in reaggregating cell culture and compared neurotrophin and LNGFR expression in developing reaggregates with that seen in the developing septum and hippocampus in situ. NGF, BDNF, NT-3 and LNGFR were each expressed in septal and hippocampal reaggregates as well as the native septum and hippocampus. Additionally, the temporal expression profiles observed in reaggregates were generally similar to those seen in the respective brain regions in situ. In order to determine whether NGF can modulate neurotrophin or LNGFR expression, reaggregates were cultured in the continual presence of either exogenous NGF or anti-NGF antibodies. NGF-treated septal cultures expressed twice the level of LNGFR mRNA as was seen in untreated septal cultures; on the other hand, septal cultures grown in the presence of anti-NGF antibodies, to neutralize endogenously synthesized NGF, displayed a 3-fold decrease in LNGFR mRNA expression compared to untreated cultures. No effects of NGF or anti-NGF were observed on LNGFR expression in hippocampal reaggregates, or on neurotrophin mRNA expression in either reaggregate type. These results suggest that regulatory mechanisms intrinsic to the septal and hippocampal regions control neurotrophin and LNGFR expression. NGF is likely to be one of these regulatory cues since it acts locally in septal reaggregates to control the developmental expression of LNGFR mRNA. The possible roles of locally synthesized NGF and other neurotrophins in the development of septal neurons are discussed.


Expression of carnosine-like immunoreactivity during retinal development in the clawed frog (Xenopus laevis).

The development of neurons immunoreactive to carnosine (beta-alanyl-L-histidine) was studied in the retina of Xenopus laevis during the premetamorphic period. Carnosine-like immunoreactivity was detected in photoreceptors from stage 39/40 (according to Nieuwkoop and Faber [Normal Tables of Xenopus laevis (Daudin), Elsevier, Amsterdam, 1956]) and in bipolar cells and their processes in the inner plexiform layer from stage 44/45. At all the developmental stages studied, neuroepithelial cells at the ciliary margin were completely unstained, suggesting that carnosine is only present in postmitotic retinal neurons. This study demonstrates a correlation between the times of appearance of carnosine-like immunoreactivity during retinal development and the onset of visual function.


Development of the brachial spinal cord in the marsupial Macropus eugenii (tammar wallaby).

The development of the brachial spinal cord was studied in the marsupial Macropus eugenii (tammar wallaby) on postnatal days 1-34. On day 1 the spinal cord was histologically immature, with a deep central canal, proliferating neuroepithelium and roof and floor plates. The lateral motor column had formed, and forelimb muscles contained primary myotubes. The spinal cord gradually attained a mature appearance between days 1-34. The results confirm the suitability of the wallaby spinal cord for studies of early mammalian development.


Chronic prenatal morphine treatment decreases G alpha s mRNA levels in neonatal frontal cortex.

G alpha s mRNA levels were measured in brain regions of newborn pups following prenatal morphine treatment. A significant decrease (24%) in G alpha s mRNA levels was observed in the frontal cortex. No changes were observed in other regions. This report demonstrates the first in vivo study of opiate effects on G-protein gene expression in neonates. The development of tolerance in vivo may involve complex interactions between several neurotransmitter systems having opposing actions on the G-protein system.


Ontogeny of calretinin in chick dorsal root ganglion neurons.

Calretinin immunostaining was performed on chick lumbosacral dorsal root ganglia during embryonic development. Calretinin-immunopositive neurons were first observed at around the 9th day of incubation. Quantitative evaluation revealed a close correlation between the number of immunopositive cells and the duration of incubation. Morphometric measurements disclosed that calretinin-immunoreactive cells belong in the large or intermediate categories of dorsal root ganglion neurons. It was concluded that the appearance of calretinin immunopositivity in spinal ganglion cells during development may be associated with both the morphological and functional maturation of this particular population of primary sensory neurons.


Characterisation of a novel glycoprotein (AvGp50) in the avian nervous system, with a monoclonal antibody.

A size fractionated lentil lectin-positive fraction derived from a deoxycholate extract of 1-day-old chick forebrain membranes was used to generate a series of monoclonal antibodies (Mabs) against neural antigens. One of these, MabSA1.7 recognises a glycoprotein which is enriched in synaptic plasma membranes, designated AvGp50. Polyacrylamide gel electrophoresis and Western blots show that AvGp50 is comprised of at least two glycoforms, with M(r)s of 53 kDa and 49 kDa respectively. AvGp50 is nervous system specific and most abundantly expressed in the forebrain, tecta and cerebellum where its pattern of expression is developmentally regulated. Immunohistochemical data localises AvGp50 to regions characterised by highly concentrated synapses, in particular, the molecular and granule cell layers of the cerebellum and in the inner and outer plexiform layers in the retina. Solubilization of the protein with the detergent Triton X-100 shows that AvGp50 is predominantly a cytoskeletally associated glycoprotein. However, when a synaptic plasma membrane fraction was treated with Triton X-114, AvGp50 partitioned into the detergent phase. Digestion of the protein with N-glycanase cleaved five N-linked carbohydrate side chains and reduced the molecular weight to approximately 34 and 31 kDa. Removal of the carbohydrate side chains led to an almost complete loss of recognition of the 34 kDa glycoform by the MabSA1.7, suggesting that the monoclonal antibody recognises a carbohydrate rather than peptide epitope.


Gliosis and ganglion cell death in the developing cat retina during hydrocephalus and after decompression.

Even after surgical decompression, infantile hydrocephalus often results in permanent neurological symptoms, including visual deficits. However, little is known about the cellular changes that may be responsible for these effects. The present study was designed to analyze the retinae of normal, mildly hydrocephalic, severely hydrocephalic and surgically decompressed kittens to determine if changes occur in the density and size of retinal ganglion cells. Hydrocephalus was induced in 10 day old kittens by intra-cisternal injection of kaolin. Kittens were allowed to survive from 7 to 28 days after injection. Animals that were decompressed received ventriculoperitoneal shunts 10-15 days after the induction of hydrocephalus and were sacrificed 10-14 days after shunt placement. The density and area of neuronal and glial cells were determined within a sample area in peripheral nasal retina. Total cell density was significantly increased in mildly and severely hydrocephalic animals but returned to normal following decompression. This change represents a significant increase in the glial population. In addition, there was a significant loss of ganglion cells in both the severely hydrocephalic and the shunted groups. Based on these findings, we conclude that gliosis occurs as a result of cell death in the retina following severe hydrocephalus, and decompression is unable to reverse these effects. Furthermore, gliosis occurs in mild cases of hydrocephalus, and may be an early indication that cellular degeneration will follow.


Developmental expression of brain derived neurotrophic factor mRNA by neurons of fetal and adult monkey prefrontal cortex.

In situ hybridization histochemistry with labeled cRNA probes complementary to monkey brain derived neurotrophic factor (BDNF) mRNAs has been used to study the cellular localization and expression of this neurotrophin in the prefrontal cerebral cortex of fetal and adult monkeys. Expression could not be detected in prefrontal cortex before the 121st fetal day. Thereafter, in fetal life and in adulthood BDNF mRNA could be detected primarily in large, putative pyramidal cells of layers III and VI throughout the prefrontal cortex. The temporal course and cellular localization of BDNF expression suggests its association with the development and stabilization of specific connections in regions of cortex that display marked functional plasticity.


Cell migration from the olfactory neuroepithelium of neonatal and adult rodents.

This study reports the presence of olfactory cell clusters in postnatal and adult animals within the lamina propria of the olfactory mucosa and the nerve fiber layer of the olfactory bulb. The results obtained from mice and rats, partially or totally unilaterally bulbectomized, have been compared with observations in intact control animals. Light microscopic observation has shown that, in bulbectomized animals, the clusters are present in both experimental and normal sides and are usually associated with olfactory axon bundles. Moreover, when compared with intact animals, differences are present in terms of number of clusters and regions from where they originate. The morphological identity of the cells of the clusters with the globose basal cell of the olfactory neuroepithelium could be demonstrated with the electron microscope. By autoradiographic means, it was possible to show that they originate from the olfactory neurogenetic matrix and migrate along olfactory axon bundles. Interestingly, the migrating cells do not express olfactory marker protein. Altogether, these observations suggest that the olfactory matrix may be capable of originating neural elements other than olfactory receptor neurons.


Developmental regulation of adenosine A1 receptors, uptake sites and endogenous adenosine in the chick retina.

Although adenosine A1 receptors mediate the inhibition of dopamine-dependent stimulation of adenylate cyclase activity in the developing chick retina, their localization and function are unknown. We have examined the localization of these receptors, and of endogenous adenosine and adenosine uptake sites at several stages of chick retinal development. A1 receptors were already localized predominantly to plexiform regions by embryonic day 12 (E12) with no gross changes at subsequent stages. Adenosine immunoreactivity was absent from retina at E8 but was detected at E12 in the ganglion cell layer, as well as cells in the inner nuclear cell layer and photoreceptors. At more advanced developmental stages the immunoreactivity was greater, but displayed similar localizations. Uptake sites labeled with [3H]nitrobenzylthioinosine (NBI) were detected even earlier using binding and autoradiographic methods. [3H]NBI binding was saturable, and Scatchard analysis demonstrated a single class of sites with a Kd of 0.91 nM and Bmax of 298 fmol/mg protein in E15 retinal membranes. The binding was displaced by unlabeled NBI and dipyridamole. NBI binding sites differentiated earlier than adenosine A1 receptors or endogenous adenosine immunoreactivity, showing a diffuse distribution at E8, but predominating in the plexiform layers of more developed retinas. The results indicate that elements of a putative purinergic system differentiate at specific localizations early in retinal development.


Expression of the barrier-associated proteins EAP-300 and claustrin in the developing central nervous system.

Immunohistochemistry of embryonic chick central nervous system (CNS) and immunocytochemistry of retinal cells were performed to compare and map the expression of two barrier-associated molecules. EAP-300 (embryonic avian polypeptide of 300 kDa) and claustrin (a 320 kDa extracellular matrix keratan sulfate proteoglycan) were both transiently expressed in CNS regions that are considered non-permissive to either neuron migration or axon growth. In the developing spinal cord, EAP-300 and claustrin were both expressed by the marginal zone early in development and by the roof plate later in embryogenesis. In the developing rhombencephalon, immunoreactivity for both molecules was also observed first in the marginal zone, and later expression was restricted mostly to the midline. In the mesencephalon, EAP-300 and claustrin were also localized to the midline, and this expression represented a continuation of the expression observed in the spinal cord roof plate and hindbrain ventral midline. In the developing retina and cerebellum, EAP-300 and claustrin were differentially expressed. In retina, EAP-300 and claustrin were expressed by Müller cells and the optic fiber layer, respectively. In cerebellum at embryonic day 12 (E12), EAP-300 was expressed by Bergman glia, but claustrin was not expressed until E15. Immunocytochemical staining of retinal and cerebellar cultures indicated that EAP-300 was expressed by a subset of radial astrocytes, as confirmed by double labeling experiments with a specific marker for radial astrocytes. These data indicate that in the absence of claustrin expression, EAP-300 was expressed specifically by radial astrocytes during developmental periods of neuron migration. Also, the coexpression of EAP-300 and claustrin in CNS regions considered to be non-permissive to neurite extension suggests that these two developmentally regulated proteins may be associated with barrier function in the developing CNS.


Effect of cocaine on electrocortical activity in fetal sheep.

The effect of cocaine on the behavioral state of six fetal sheep was studied during gestational ages between 128-135 days. Two to eight days after surgery, fetuses received either a continuous 60 min intravenous infusion of cocaine HCl (33.4 mg) or saline. The infusions were preceded and followed by control periods of 102 min. Cocaine induced a disruption in fetal behavioral state cyclicity and a decrease in the amount of time spent in rapid eye movement sleep (P < 0.01) and non-rapid eye movement sleep (P < 0.05) during the infusion, but not during the recovery period. Spectral amplitude of the electrocortical activity at all three cortical locations increased within most one-third octave bands between 0.8-4 Hz and decreased within most bands between 16-25 Hz (P < 0.05) compared to controls. There were no differences in spectral amplitude between pre- and post-cocaine periods at any location over the 25 frequency bands studied (P > 0.6) except for one frequency band centered at 12.5 Hz. The effects of a one hour cocaine infusion on fetal cortical electrical activity are diffuse, but short-lived, and occur independently of changes in fetal oxygenation.


Gallstone-associated acute pancreatitis: a field for endoscopic therapy?

The pathogenesis of acute pancreatitis even if associated with gallstone remains unclear. This explains that the best treatment for gallstone-associated pancreatitis remains a matter of debate. However, different theoretical and clinical evaluations support the strategy to perform an endoscopic papillotomy if a common bile duct stone is suspected and the course of the disease does not improve spontaneously during the first 48 h. The discrepancies of the current data and the difficulties to give rational suggestions have been reviewed.


Relevant aspects of physiology in chronic pancreatitis.

Specific points of physiology which are relevant for the understanding and management of chronic pancreatitis are focussed on. First, the regulatory factors of exocrine pancreatic secretion are described, discussing the role of the cholinergic system and of the classical gut hormones cholecystokinin and secretin. The association of upper gastrointestinal dysfunctions with a disturbed exocrine pancreatic response is also dealt with. Lastly, these changes are related to clinical consequences such as maldigestion or malabsorption.


Role of imaging technics in the classification of acute pancreatitis.

This paper is a review of some aspects of acute pancreatitis (AP) (definition criteria, outcome, and prognostic factors) and a reminder of the invaluable contribution of computed tomography (CT) in confirming the diagnosis of AP, distinguishing between edematous and necrotizing forms, and in providing prognostic information via detection of possible extrapancreatic spreads (EPS). The Geneva experience (510 cases of AP over a 9-year period) has shown that no fatalities occurred when no EPS were found on CT performed within 48 h of admission; mortality was 1.5% in the case of 1 or 2 EPS and 19% in the case of 3 EPS or more.


Etiology and natural history of chronic pancreatitis.

Chronic pancreatitis (CP) is a rare but serious disease with high morbidity and mortality. Its exact etiology remains uncertain, but several associated conditions have been identified. Geographical distribution of CP can be linked to alcoholism, especially in countries with high-protein, high-fat diets. In Afro-Asiatic countries with protein malnutrition, however, CP is frequently observed in children and young nonalcoholic adults from the poorest segments of these societies. To analyze the natural history of CP, besides etiology three additional main factors have to be considered, namely clinical pattern and therapy (surgery), pancreatic function (endocrine and exocrine), and morphology. During progress of the disease clinical picture, morphology and pancreatic function have typical correlative changes. Basically, from this viewpoint, three typical models of the disease can be distinguished: (1) early stage of uncomplicated CP; (2) late stage of uncomplicated CP, and (3) complicated CP, a disease stage which is characterised by local complications (chiefly pseudocysts and duct obstruction). The main concept of the natural history of CP bases on the thesis that CP burns itself out with spontaneous relief of pain, i.e. persistent freedom from pain occurs parallel with severe pancreatic dysfunction in the late stage of the disease. In the clinical picture and long-term course, nonalcoholic CP differs in certain essential respects from alcoholic CP; however, the two forms do not differ essentially as regards mortality and survival.


Surgical treatment of painful chronic pancreatitis: an unresolved problem?

Relief of pain in chronic pancreatitis is the major problem warranting surgical treatment in this disease. The mechanism of pain is largely unknown and several types of operation have been devised for treatment. Side-to-side pancreaticojejunostomy (Partington-Rochelle) and pancreaticoduodenectomy according to Whipple have stood the test of time. Recently, new surgical options have been explored like the operation according to Beger, segmental autotransplantation, and duodenum-preserving total pancreatectomy. Because of the reluctance to refer this type patient for surgery, treatment with analgesic drugs is continued for quite some time and once analgesia addiction has developed clinical judgement in these patients is severely hampered. Surgery can be performed with 70-80% success and with limited morbidity as well as low mortality. For these reasons surgery should be discussed early in the disease if pain becomes a major problem. If these patients are operated prior to analgesia addiction, maybe the long-term prognosis will improve. The diagnostic and surgical approach will be discussed in detail with a plea for considering surgery early in the course of disease.


Acute pancreatitis: when and how to operate.

Patients with proved necrotizing pancreatitis should be treated in an intensive care unit. Surgical management of necrotizing pancreatitis is indicated if an acute abdomen or persistent or increasing signs of organ complications develop, such as pulmonary or renal insufficiency, cardiocirculatory dysfunction or metabolic disorders, and these do not respond to maximum intensive care treatment over at least 72 h. Besides these so-called non-responders to ICU treatment, operative management is clearly indicated in patients who develop signs of sepsis on the basis of a bacteriologically positive fine-needle aspiration of pancreatic necroses. In patients with minor necroses without any bacterial contamination and without extensive retroperitoneal fatty tissue necroses intensive care therapy can be successful without the necessity of a surgical intervention. The gold standard of surgical management of necrotizing pancreatitis is careful removal of necrotic tissue, drainage of bacterially infected area, elimination of the pancreatogenic ascites in order to prevent systemic spread of vasoactive and toxic substances and interruption of the inflammatory process. For the treatment of pancreatic necrosis we strongly support surgical debridement (necrosectomy), supplemented by postoperative closed continuous lavage of the lesser sac and the adjacent necrotic cavities. In 152 patients suffering from severe necrotizing pancreatitis the hospital mortality was 12.5% (19/152) by this surgical approach.


Bradycardia during reversible hypovolaemic shock: associated neural reflex mechanisms and clinical implications.

1. Heart rate response to reversible central hypovolaemia can be divided into three stages. In the first stage (corresponding to a reduction of the blood volume by approximately 15%) a modest increase in heart rate (< 100 beats/min) and total peripheral resistance compensate for the blood loss, and a near normal arterial blood pressure prevails (preshock). During the second stage, a reduction of the central blood volume by approximately 30% results in a decrease in heart rate, total peripheral resistance and blood pressure due to activation of unmyelinated vagal afferents (C-fibres) from the left ventricle. In the third stage, blood pressure falls further as haemorrhage continues and tachycardia (> 120 beats/min) is manifest. This stage may proceed into irreversible shock with death from cardiac arrest probably related to the formation of free oxygen radicals. 2. Recognition of the vasodepressor-cardioinhibitory reaction to a reduced circulating blood volume is important and suggests the need for immediate treatment with volume expansion in critically ill patients.


Afferent control of vasopressin and renin release during haemorrhage in normal and autonomically blocked rabbits.

1. The role of the arterial and cardiac baroreceptors on the arginine vasopressin (AVP) and plasma renin activity (PRA) responses to haemorrhage was studied in conscious rabbits. They were bled at a rate of approximately 3% of their blood volume (BV)/min, both when the autonomic nervous system (ANS) was intact and during ANS blockade, which markedly enhances the AVP response due to the much greater haemodynamic disturbance. Under each condition of ANS function 2 x 2 factorial analysis was performed, each with four groups of rabbits, including animals with both sets of baroreceptors working, one or other set working and neither set working. 2. With intact ANS, haemorrhage had to be terminated at different times in the four groups. This presents problems for factorial analysis due to differences in the relationship between plasma AVP (or PRA) and release rate. A method for overcoming this was developed by extrapolating the BV-log AVP curves to a common time from the start of bleeding. 3. Under both conditions of ANS function the arterial and cardiac baroreceptors together accounted for 90-95% of the rise in AVP during haemorrhage. With normal ANS function, the rise in AVP was about 70% through cardiac (probably ventricular) baroreceptors (P = 0.01) and about 30% through arterial baroreceptors (P = 0.08). This compares with an earlier study at a rate of bleeding of 1.8% BV/min, where the entire drive came from the cardiac receptors. During ANS blockade, plasma AVP was enhanced approximately five-fold, which was mostly mediated through the arterial baroreceptors, but the cardiac baroreceptor component was also greater; arterial/cardiac baroreceptor drive was 2/1. 4. Baroreflexes played no role in renin release during haemorrhage, but the experiments with ANS blockade suggest that a hormonal factor, which was related to the cardiac innervation, may limit the rise in PRA in the latter part of haemorrhage.


Arterial baroreflex control of heart period is not related to blood pressure variability in conscious hypertensive and normotensive rats.

1. The short-term (within 30 min periods) and the long-term (among 30 min periods) variabilities, expressed as variation coefficients, of blood pressure (BP) and heart period (HP) were studied using a computer analysis of BP recordings in freely moving genetically hypertensive (LH), normotensive (LN) and low BP (LL) rats of Lyon strains at ages 5, 9, 21 and 40 weeks. The baroreflex control of HP was estimated with the slope of the linear relationship between systolic BP (SBP) and HP (SBP-HP slope) computed after phenylephrine and nitroglycerin injections. 2. Short-term variability of BP increased between 5 and 9 weeks of age and then remained stable. Hypertension was accompanied by an increase in both short- and long-term variabilities of diastolic BP in adult rats. 3. A sharp increase in SBP-HP slope was observed between 5 and 9 weeks of age in LN rats. SBP-HP slope of LH rats increased slightly up to 21 weeks but remained lower than that of normotensive controls. 4. The weak inverse correlation existing between SBP-HP slope and BP variability appeared to be mediated by the BP level. In addition, atropine which is known to abolish almost completely the SBP-HP slope, did not increase BP variability. It is concluded that SBP-HP slope is not linearly associated with BP variability in conscious rats.


Effects of 4 weeks endurance training on cardiac left ventricular structure and function.

1. The effect of 4 weeks of moderate exercise training on cardiac left ventricular structure and function was examined by repeated echocardiographic/Doppler examination in 10 previously sedentary volunteers participating in a single blind, sedentary phase controlled, cross-over study. 2. Left ventricular internal diastolic diameter increased after 4 weeks of training from 4.98 to 5.11 cm with a further increase to 5.24 cm after 2 weeks of detraining (s.e.d. 0.05 cm, P < 0.01). These increases were still significant after adjustment for heart rate which fell from a pretraining average of 67.0-59.9 beats/min after 4 weeks of training. 3. There were no significant changes in systolic diameter or function and left ventricular wall thicknesses were unchanged during training, but were thinned after 2 weeks subsequent detraining. 4. The ratio of early to late transmitral filling velocity (E/A ratio) was significantly increased by training. Although E/A ratio was shown to be heart rate sensitive, training effects were still evident after adjustment for heart rate. 5. These changes in left ventricular volumes and function may contribute to the cardiovascular reflex changes previously shown to be produced by identical training programmes.


Epithelial disruption by proteases augments the responsiveness of porcine bronchial segments.

1. The effect of disruption to the epithelium of intact porcine bronchi was examined by comparing the responsiveness to agonists applied to the adventitial and luminal surfaces. The development of smooth muscle tone was measured as an increase in pressure in an isovolumic bronchial segment of approximately 2 mm i.d. The reactivity and sensitivity to acetylcholine (ACh) introduced intraluminally was greatly attenuated when compared with adventitial addition. 2. Luminal exposure to K+ and vanadate (VO3-) had little effect compared with strong responses obtained by adventitial application. 3. Intraluminal exposure to trypsin, chymotrypsin and elastase (1 mg/mL) selectively augmented both the sensitivity and the reactivity to the luminal addition of ACh, K+ and carbachol. 4. Mechanical removal of the epithelium produced a 716-fold increase in sensitivity to ACh introduced luminally but had no effect on ACh applied adventitially. 5. The inhibitory effects of luminally introduced isoprenaline on electrical field stimulation responses were also significantly potentiated in segments stripped of epithelium. 6. The evidence presented here indicates that the responsiveness of in vitro airways segments is highly influenced by the epithelial layer, which acts most prominently as a barrier inhibiting the penetration of luminally introduced agonists.


Potassium accelerates urinary sodium excretion during salt loading without stimulating atrial natriuretic polypeptide secretion.

1. Effects of potassium (K) supplementation (100 mEq/day) on urinary sodium (Na) excretion and on the secretion of atrial natriuretic polypeptide (ANP) during salt loading (350 mEq/day) were studied in 12 healthy salt-resistant normotensives under strictly controlled metabolic ward conditions. 2. Urinary volume and Na excretion on the first day of the high salt period (HSP) were significantly greater in the K-supplemented group (KG) than in the control group (CG). 3. There was a significant gain in bodyweight after salt loading in both groups, with a significantly greater gain in CG on the second day of HSP. Haematocrit decreased significantly during salt loading in both groups, the degree of which was significantly greater in CG. 4. Plasma norepinephrine decreased significantly during salt loading in both groups, the degree of which was significantly less in KG than in CG. A significant increase in plasma ANP was observed in CG on and after the second day of HSP, while a significant increase in plasma ANP was observed on the fifth day of HSP in KG. 5. These findings indicate that K supplementation accelerates diuresis and natriuresis, resulting in moderate suppression of volume expansion induced by salt loading and that this accelerated diuresis and natriuresis is not a result of the action of ANP.


Influence of cimetidine on gallamine-induced neuromuscular paralysis in rats.

1. The effect of cimetidine on gallamine-induced neuromuscular paralysis was evaluated in urethane-anaesthetized and mechanically ventilated rats. 2. Intravenous cimetidine (between 3.2 and 100 mg/kg) produced an immediate and sustained potentiation of gallamine-induced neuromuscular paralysis. 3. Intravenous calcium (0.35 mmol/kg) completely reversed the cimetidine-induced potentiation of gallamine's effect. 4. Cimetidine alone did not produce neuromuscular paralysis. 5. Cimetidine did not alter either the serum concentration or the total serum clearance of gallamine. 6. Cimetidine thus potentiates the action of gallamine via a pharmacodynamic and not a pharmacokinetic mechanism.


Direct renal effects of endothelin in chronic hypoxic spontaneously hypertensive rats.

1. The direct renal effects of endothelin (ET) were studied in eight chronic hypoxic rats (HA) and eight sea level (SL) spontaneously hypertensive rats (SHR). 2. After 4 weeks of exposure to simulated 5486 m (18,000 ft) hypoxia, all HA rats were in apparently good health, and baseline renal function, except effective renal blood flow, was not significantly different from SL rats. 3. Intrarenal arterial administration of ET (600 ng/kg per h) reduced ipsilateral renal excretion of water, sodium and potassium, glomerular filtration rate and effective renal plasma flow in both SL and HA rats to almost the same extent. 4. Administration of ET antiserum, however, increased the renal excretion of water in HA rats. 5. It is concluded that ET may play a role in the renal regulation of chronic hypoxic SHR.


Assessing vascular reactivity of arteries in the small vessel myograph.

1. Using a small vessel myograph, experiments were performed on rat small mesenteric arteries (and in some cases dog small coronary arteries) to assess the dependence of vasoconstrictor potency (EC50) and maximum response (Emax) on the initial passive conditions and on the mode of recording (i.e. isometric, isobaric or isotonic). 2. Maximum active isometric tension development to methoxamine occurred at different points on the passive diameter--tension curve depending on the origin of the vessel. The point of maximum sensitivity of methoxamine did not coincide with the point of maximum tension development on the passive diameter-tension curve. 3. Vascular reactivity to methoxamine was assessed under isobaric, isotonic and isometric conditions using a new computerized myograph. Methoxamine was significantly more potent, but only by a factor of twofold, when assessed under isometric conditions. In addition, the maximum response to methoxamine, in terms of diameter change, was always greater under isobaric than under isotonic conditions. 4. The results show that, in studies comparing vascular reactivity of vasoactive drugs, the results depend, to some extent, on the initial passive conditions selected. In terms of assessing the pharmacological activity of drugs on isolated blood vessels, the use of common isometric recording procedures are adequate. However, the use of isobaric, isotonic and isometric recording procedures have shown the complexities of vascular reactivity which depend on the passive and active properties of the blood vessel. These factors may need to be taken into account when comparing the reactivity of isolated blood vessels.


The role of the renal nerves in renin synthesis.

1. Renin synthesis and secretion were studied in Balb/c mice with a denervated left kidney. 2. Denervation inhibited renin secretion. 3. Denervation reduced the renal renin content. 4. Denervation reduced renal renin mRNA. 5. Renal denervation inhibits renin secretion by blocking the synthetic system prior to mRNA formation.


Effects of noise on blood pressure and vascular reactivities.

1. The present study investigated the effects of noise stress (100 dB, 1 kHz, 4 h/day, 6 days/week) on both systolic blood pressure and vascular reactivities in rats exposed to noise for 2 and 4 weeks. The systolic blood pressure was significantly increased after rats were exposed to noise for 4 weeks but not 2 weeks. 2. In isolated thoracic aortic rings, the responses to serotonin were enhanced in noise-treated rats while there were no changes in response to phenylephrine and high K+ between noise-treated and control rats. 3. The relaxant responses to endothelium-dependent vasodilators (A23187 and acetylcholine) in noise-treated rats were less than those in control rats. The vasodilator responses to acetylcholine were completely abolished by methylene blue or N omega-nitro-L-arginine. 4. Responses to the endothelium-independent vasodilator, nitroglycerin, were not significantly changed after rats were exposed to noise. 5. The enhanced response to vasoconstrictors and the attenuation to endothelium-dependent vasodilators may account for elevations in blood pressure during noise stress. This indicates that the elevation in blood pressure by noise stress may be partly due to the deterioration of endothelial function.


Cholesterol and lipoproteins: beyond atherogenesis.

Although much emphasis has been placed on the role of cholesterol and lipoproteins in atherosclerotic plaque formation, recent studies suggest that lipids have other vascular actions which may contribute to the pathogenesis of myocardial ischemia. These include deleterious effects of lipids on platelet and endothelial cell function, coagulation, fibrinolysis, and prostacyclin metabolism. The purpose of this report is to review recent data regarding the nonatherogenic effects of lipids and provide insight as to how lipid lowering might contribute to clinically important improvements in vascular biology.


Early and late potentials in postinfarction patients.

Injury within the myocardium may cause disturbances of the intraventricular conduction and may be manifested by altered spectrum of the QRS complex. The purpose of this report is to indicate the existence of microvolt early waves appearing at the beginning of the QRS complex in some postinfarction patients with low ejection fraction. The method used incorporated the high resolution ECG recording, where the "pyramid" electrode location was applied. Results indicated a relationship between high frequency potentials, QRS duration time, and ejection fraction, as well as dependence between ventricular tachycardia/fibrillation (VT/VF) occurrence and early and/or late ventricular activity. In patients with secondary VT/VF where early and late potentials were found, the ejection fraction (EF) was the lowest (26 +/- 15%). Early waves appeared more often in patients with secondary VT/VF (30%) compared with those with primary VT/VF (8%). In patients without VT/VF and without myocardial infarction (MI) early potentials were not remarked. In those patients late potentials also appeared less often. It could be concluded that existence of early potentials, late activity, and significant prolonged total ventricular activation time is associated with wide dispersion and nonuniform delay of the electrical activity within the myocardium as well as with serious lowering of the ejection fraction. Therefore, existence of early waves seems to be a significant diagnostic factor for the poor hemodynamic condition as well as a serious warning for the recurrent late ventricular tachycardia/fibrillation.


Combination of ramipril and hydrochlorothiazide in the treatment of mild to moderate hypertension: Part 1--A double-blind, comparative, multicenter study in nonresponders to ramipril monotherapy.

In a parallel-group multicenter study, the efficacy and safety of combination therapy with ramipril 5 mg plus hydrochlorothiazide 25 mg were compared double-blind with those of 5 mg and 10 mg ramipril monotherapy in patients with mild to moderate hypertension who had not responded adequately to ramipril 5 mg alone. Patients were initially treated single-blind for 1 week with ramipril 2.5 mg and 3 weeks with ramipril 5 mg. Of 240 patients enrolled, 165 were subsequently classed as nonresponders (diastolic blood pressure > 90 mmHg) and were randomized to one of the three double-blind treatments for a further 4 weeks. In the double-blind phase, the mean reductions in supine systolic and diastolic blood pressures at end point were significantly greater in the 5 mg plus 25 mg combination group (11.6/10.6 mmHg) than in the groups receiving ramipril 5 mg (6.2/5.9 mmHg; both p < 0.01) and ramipril 10 mg (7.4/7.1 mmHg; both p < 0.05). The proportion of responders at end point was also higher for combination therapy (72%) than for monotherapy (48% for ramipril 5 mg and 62% for ramipril 10 mg). All three treatments were well tolerated. Analysis of laboratory values revealed no clinically important changes.


Influence of atenolol on the relationship between heart rate and QT interval in patients with exercise-induced myocardial ischemia.

The aim of this study was to analyze the relationship between heart rate and QT interval (HR-QT) during exercise in control subjects (Group A) and in patients with coronary artery disease (CAD) with effort angina and without previous myocardial infarction (MI) (Group B). The diagnosis of CAD was confirmed by coronarographic examination. The correlation HR-QT was significant (p < 0.001) in both groups on effort and at recovery. The analysis of the regression HR-QT was carried out separately, both on effort in upright position and at rest in supine position, to avoid the influence of posture on QT length. During effort, the regression line showed lower slope and intercept values in Group B (p < 0.001) than those for Group A. A similar behavior was also observed at rest. Thus, at the highest heart rate, where ECG signs of ischemia (ST depression > 1 mm) frequently occurred, a longer QT interval was present in Group B. Moreover, in Group B, the QT interval in the presence of ECG signs of ischemia was significantly longer (p < 0.01) than in Group A at comparable heart rates both on effort and at rest, thereby confirming the result obtained by comparing both regression lines. The same effort protocol was repeated in Group B patients after acute administration of atenolol 100 mg per os. After atenolol administration, the analysis of the regression HR-QT in Group B clearly showed a shorter QT interval than that obtained in washout period during the baseline test at the highest heart rates where the ECG frequently showed signs of ischemia.(ABSTRACT TRUNCATED AT 250 WORDS)


The effect of chronic atrial overdrive suppression pacing on the incidence of supraventricular tachyarrhythmias.

Chronic overdrive suppression pacing has been suggested as an effective adjunctive method for reducing the incidence of cardiac tachyarrhythmias. Documentation of effectiveness during prolonged monitoring is lacking, however. To assess more accurately the long-term utility of this treatment modality for medically refractory supraventricular tachyarrhythmias (SVTs), 10 patients with atrially implanted Intermedics Intertach pacemakers were randomly assigned to either a low or a high bradycardia (back-up) pacing rate. SVT counts were performed during matching follow-up periods both at the initial rate and after rate crossover. The primary antitachycardia modality of this pacemaker (P mod) provides burst pacing to terminate tachycardia episodes, and P mod counters were utilized to quantitate SVT episodes. Tachycardia termination algorithms were programmed to "no restart" and were not changed during the study. The P mod use counter, therefore, reflected the number of discrete episodes of SVTs. Pacemaker implantation diagnoses include atrial flutter, concealed bypass tract, AV nodal reentry, intraatrial reentry, and Wolff-Parkinson-White associated tachycardia. Patient age was 59 +/- 18 yrs. The average pacemaker back-up low rate was 45.7 +/- 4 versus a back-up high rate of 85.1 +/- 2 beats/min. Follow-up was for 57.4 days +/- 33 days at the low rate and 57.3 days +/- 34 days at the high rate (r = 0.99). There was no difference in SVT incidence with a P mod usage of 98.4 +/- 106 at the low rate and 100.8 +/- 94 at the high rate (p = NS). In this blinded, randomized cross-over trial, chronic atrial overdrive suppression pacing did not reduce the overall incidence of SVT episodes during prolonged monitoring.


Embolus in transit.

Since the advent of echocardiography, embolus in transit, historically found during surgical exploration or on postmortem examination, has been found with increasing frequency on antemortem examination. There is an inherent high mortality rate with this condition and awareness of the association between deep venous thrombosis and embolus in transit is paramount. On echocardiography the embolus is typically seen as a pleomorphic mass moving in a tumbling fashion. The most frequent symptoms are dyspnea and near syncopal episodes. The most common signs are diastolic "tumor plop" and a systolic ejection murmur heard on auscultation. Despite the success of some medical interventions, surgery should be strongly considered in patients with embolus in transit.


Acute ischemic exacerbation of rheumatic mitral regurgitation.

This report describes a patient who developed acute severe exacerbation of mild rheumatic mitral regurgitation caused by ischemia in the territory of a small, non-dominant circumflex coronary artery without myocardial infarction.


Adult onset Niemann-Pick disease type C presenting with dementia and absent organomegaly.

A 39-year-old female presented to the Bryan Memory Disorders Clinic at Duke University with a 7-year history of an atypical progressive dementia, mildly impaired vertical gaze, dysarthria and mild ataxia. There was no evidence of organomegaly by clinical examination or by radionuclide liver/spleen scan. Brain biopsy disclosed a neuronal storage disorder characterized by ballooned neurons filled with oligo-lamellar cytosomes and lipid droplets. Cultured skin fibroblasts had diminished sphingomyelinase activity and impaired cholesterol esterification, although peripheral leukocyte sphingomyelinase activity was normal. Two years after biopsy, follow-up examination revealed marked progression of vertical gaze paralysis and ataxia. This case expands the clinical spectrum of Niemann-Pick disease type C by presenting in adulthood with subtle neurologic abnormalities; no visceromegaly and profound dementia.


Immunohistochemical characterization of pineocytomas.

The immunoreactivity of a panel of poly- and monoclonal antibodies raised against different neuronal and glial antigens, was studied in paraffin-embedded specimens of five pineocytomas. Antibodies against neuron-specific enolase (NSE), S-100 protein, neurofilaments protein (NFP), glial fibrillar acidic protein (GFAP), myelin basic protein (MBP), Synaptophysin (SYN) and vimentin were used. NSE immunoreactivity was detectable in nearly all tumoral cells. S-100 expression was present in all tumors, but a significant variation in the number of S-100 positive cells was noted in the different specimens; they ranged from 3% of S-100 positive cells in some areas of case 1, to 30% in other areas of case 4. The number of GFAP-positive cells detected in four of the five tumors was minimal. In the remaining tumor, 10% to 14% the positively GFAP antisera-stained cells could be seen in some areas; this tumor was considered a pineocytoma with astrocytic differentiation. MBP immunoreactivity was found in isolated cells only in one tumor. Neurofilaments or vimentin expression could not be found in tumoral cells. Synaptophysin was studied in two tumors (cases 4 and 5). Both tumors showed a fine granular neuropil pattern of immunoreactivity, but only in isolated cells of case 5, occasional cytoplasmic positivity could be determined. Based upon our results we can conclude that NSE and synaptophysin are markers that may be applied in the diagnosis of pineocytomas. Further, our results support the hypothesis that the tumoral cells in these neoplasms are of neuroendocrine origin.


Autonomic function testing in diabetic subjects using sequential measurements.

The repeatability of commonly used electrocardiographic (ECG) related autonomic function tests was investigated in 25 diabetic subjects, mean age 44 (range 18-67) years and mean duration of diabetes 10 (< 1-35) years. Tests were based on deep breathing, the Valsalva manoeuvre, relaxed normal breathing, and standing up from a lying position. All tests were repeated twice at each session, with two sessions separated by a mean of 5 (range 3-8) months. ECG and respiratory data were recorded for subsequent analysis. Maximum (max) and minimum (min) RR intervals and instantaneous heart rate (HR) were determined. The mean and repeatability data (within-subject standard deviation) for 21 different test results were calculated, and included deep breathing sitting (max-min)RR 226 +/- 39 ms and (max-min)HR 19.3 +/- 3.2 beats min-1, Valsalva manoeuvre 1.74 +/- 0.18, and lying-to-standing RR ratio 1.19 +/- 0.07. The within-subject repeatability to between-subject variability ratios consistently demonstrated that it is better to perform the deep breathing test sitting (with the smaller mean ratio of 0.37) rather than supine (0.46), with the use of three respiratory cycles rather than a single cycle (0.33 compared with 0.50 for (max-min)RR). A significant (p = 0.037) relationship was found between variability in heart rate changes and variability in air volume breathed during the deep breathing test.


Progressive supranuclear palsy with widespread cerebral lesions.

A 51-year-old woman with no history of any familial neurological diseases initially presented with numbness in her extremities, slowing of movements, comprehension deficit, memory disturbance, dyscalculia, muscle rigidity, hyperreflexia, Parkinsonian gait, increasing disorientation, left-right disturbance, finger agnosia, alexia, acalculia, apraxia, aspontaneity, euphoria, gait disturbance, aphasia, echolalia, and in the terminal stage, mutism, contracture of lower extremities and cachexia. She died of bronchopneumonia at the age of 55. The brain showed widespread cerebral lesions, consisting of nerve cell loss and neurofibrillary tangles in the frontal, parietal and occipital cortex, demyelination and gliosis in the frontal, parietal and occipital subcortical white matter in addition to the typical pathological findings of progressive supranuclear palsy (PSP): severe neuronal loss with gliosis and neurofibrillary tangles (NFTs) in the subthalamic nucleus, globus pallidus and substantia nigra. In conclusion, we present a case of PSP with unusual clinical features (extrapyramidal signs, frontal and parietal lobe syndromes without ophthalmoplegia) and neuropathologically widespread cerebral lesions in addition to the typical pathological findings of PSP. The differential diagnosis of PSP and Alzheimer's disease and other degenerative disorders is discussed.


Chronic fatigue syndrome: studies on skeletal muscle.

Chronic fatigue syndrome represents a poorly defined disease with protean clinical manifestations, the majority of them expressed as a muscle fatigue or as inability to maintain the expected muscle strength. In the present work we studied muscle function and muscle histopathology in 20 patients fulfilling the proposed criteria for chronic fatigue syndrome. Special interest is directed towards the immunoreactive expression of class I MHC molecules comparing some inflammatory and virus-related myopathies with muscles from chronic fatigue syndrome. Only minor morphological changes were detected in 9 out of 20 patients of the series. The nonspecific morphological changes in muscle tissue and the lack of class I MHC expression does not support the viral etiology of muscle fatigue in chronic fatigue syndrome. In contrast with the reported clinical improvement with high doses of essential fatty acids, our patients' clinical condition did not improve after three months of L-carnitine therapy.


Prevalence of cardiovascular autonomic dysfunction assessed by spectral analysis, vector analysis, and standard tests of heart rate variation and blood pressure responses at various stages of diabetic neuropathy.

To establish a test battery for the detection and characterization of cardiovascular autonomic neuropathy (CADN) and to evaluate its prevalence, a number of autonomic function tests based on spectral analysis, vector analysis, and standard tests of heart rate variation and blood pressure responses were performed in 261 diabetic patients aged 11-76 years with various stages of peripheral neuropathy. The percentages of abnormal results in the individual tests based on heart rate variation were 6-31% in 115 patients without peripheral neuropathy, 16-45% in 61 patients with subclinical neuropathy, 22-59% in 73 patients with symptomatic peripheral neuropathy, and 67-100% in 12 patients with the latter in conjunction with autonomic symptoms (p < 0.05). The most frequently abnormal indices, each representing a different physiological basis, were the coefficient of variation, low-frequency and mid-frequency power spectrum at rest, mean circular resultant, postural change in systolic blood pressure, and, in particular, the max/min 30:15 ratio and Valsalva ratio. CADN, defined as the presence of > or = 3 abnormalities among these seven parameters was detected in none of 120 control subjects, 13.0% of the patients without peripheral neuropathy, 34.4% of those with subclinical neuropathy, 49.3% of those with symptomatic peripheral neuropathy, and in 100% of the subjects with the latter and concomitant autonomic symptoms (p < 0.05). The overall prevalence of CADN in 103 patients completing all parameters was 46.6%. The corresponding rate of CADN defined as > or = 2 abnormalities among the five tests included in an optimized version of the battery proposed by Ewing and Clarke was 38.8%.(ABSTRACT TRUNCATED AT 250 WORDS)


Sensory-motor chronic neuropathy in two siblings: atypical presentation of tomaculous neuropathy.

Two siblings, born from a consanguineous marriage, in which tomaculous neuropathy (TN), diagnosed after nerve biopsy, presented as a chronic sensory-motor neuropathy. The clinical manifestations include also in both patients thyroid multinodular struma and in one case myopathic-like features. We believe that TN must be suspected not only in hereditary neuropathy with liability to pressure palsies, but also in any case with chronic sensory-motor neuropathy.


Feeding in infancy and the risk of type 1 diabetes mellitus in Finnish children. The 'Childhood Diabetes in Finland' Study Group.

In a case-control design the feeding in infancy of newly diagnosed 7- to 14-year-old diabetic children (n = 426) was compared with that of age- and sex-matched non-diabetic children (n = 426) randomly selected from the Finnish population registry. All 7- to 14-year-old diabetic children diagnosed from September 1986 to the end of April 1989 from all hospitals which treat diabetic children in Finland were invited to participate in the study. Breast-feeding was initiated in almost all children, but during the birth years of this study population (1972-1982), an increase was observed in the duration of breast-feeding (whether alone or in combination with supplementary feeding) and in the age of introduction of supplementary milk feeding. The risk of Type 1 diabetes was decreased in the children who were totally breast-fed for at least 2 months (odds ratio (OR) 0.64, 95% confidence interval (CI) 0.42-0.98) or 3 months (OR 0.67, 95% CI 0.48-0.95) or exclusively breast-fed for at least 2 months (OR 0.60, 95% CI 0.41-0.89) or 3 months (OR 0.63, 95% CI 0.43-0.93). Those children who were younger than 2 months (OR 1.54, 95% CI 1.08-2.18) or 3 months (OR 1.52, 95% CI 1.11-2.08) at the time when supplementary milk feeding was begun had an increased risk of Type 1 diabetes. These associations remained significant after adjusting for the mother's education. The results suggest that early infant feeding patterns are associated with the risk of Type 1 diabetes developing at the age of 7 to 14 years.


High prevalence of gestational diabetes in women from ethnic minority groups.

The influence of ethnic origin, body mass index, and parity on the frequency of gestational diabetes was assessed in 11,205 consecutive women attending a multiracial antenatal clinic in London, where all women were screened for gestational diabetes. Logistic regression was used to model the relationship between gestational diabetes and ethnic origin, age, body mass index (BMI), and parity. Results were presented as adjusted odds ratios, where the reference categories are White women, age < 25 years, BMI < 27, and parity < 3. Ethnic origin was the dominant influence on the prevalence of gestational diabetes. Women from ethnic groups other than White had a higher frequency of gestational diabetes than White women (2.9% vs 0.4%, p < 0.001). Compared to White women the relative risk of gestational diabetes in the other ethnic groups was: Black 3.1 (95% confidence limits 1.8-5.5), South East Asian 7.6 (4.1-14.1), Indian 11.3 (6.8-18.8), and miscellaneous 5.9 (3.5-9.9). Increasing age was an independent risk factor. The relative risk was higher in women > or = 35 years in all ethnic groups other than in South East Asian women. Obesity (BMI > or = 27) was a further independent risk factor in all ethnic groups except in the Indian and South East Asian women. Parity > or = 3 increased the relative risk of gestational diabetes in the White, Black, and South East Asian women only.(ABSTRACT TRUNCATED AT 250 WORDS)


Morning versus bedtime isophane insulin in type 2 (non-insulin dependent) diabetes mellitus.

Morning versus bedtime administration of NPH insulin was compared in 12 subjects with Type 2 diabetes and overt fasting hyperglycaemia. Subjects were studied at baseline (diet alone) and after 2 months on each of the two insulin programmes in a random crossover design, in which dosage was increased until at least one daily preprandial blood glucose was consistently in the range of 3.9 to 6.0 mmol l-1. Mean (+/- SEM) daily total insulin dosage was equivalent for the morning (0.36 +/- 0.03 units kg-1) and for the bedtime (0.37 +/- 0.03 units kg-1) insulin administration schedules. Glycaemic control was improved on both insulin regimens, but was better on bedtime than morning insulin. Fasting plasma glucose (mmol l-1) was 12.0 +/- 0.7 (baseline), 8.6 +/- 0.7 (morning), and 4.6 +/- 0.3 (bedtime), respectively. Mean 24 h plasma glucose (mmol l-1) was 13.3 +/- 1.3, 9.0 +/- 0.7, and 7.8 +/- 0.7. Glycated haemoglobin (%) was 7.65 +/- 0.35, 6.23 +/- 0.26, and 5.81 +/- 0.32. The improvement of basal glycaemia is a consequence of increased basal metabolic clearance of glucose (baseline, 47.6 +/- 3.1 ml m-2 min-1; morning 63.5 +/- 5.4, bedtime 103.5 +/- 7.1). There was no change in hepatic glucose output. It is concluded that bedtime administration of intermediate acting insulin results in increased basal insulinaemia, leading to improved basal glycaemia and consequent improved overall metabolic control, compared to morning insulin administration. Therefore, bedtime may be the preferable timing of insulin therapy for patients with Type 2 diabetes and overt fasting hyperglycaemia.


Relationship between insulin injection regimen and metabolic control in young Danish type 1 diabetic patients. The Danish Study Group of Diabetes in Childhood.

In 1987 and 1989 nationwide screening for HbA1c was carried out in Denmark. Twenty-one paediatric departments treating children with diabetes participated in the first study and twenty-two in the second. During this 2-year period metabolic control deteriorated despite the fact that the use of multiple injection regimens increased from 39% to 54%. The possible reasons for the deterioration in metabolic control were examined in the 429 children (> or = 9 years, 1987) and adolescents (< 18.8 years, 1989) who participated in both studies. All had diabetes duration of greater than one year (1987) and all were treated by the same departments during the study period. The children were divided into three subgroups according to injection regimen. Group A (n = 128) received twice-daily insulin injections; group B (n = 171) were on multiple injections (three or more) while group C (n = 130) shifted from twice-daily insulin to multiple injections during the 2-year period. A deterioration of blood glucose control as assessed by HbA1c was observed in all three treatment groups. For group C the 2-year increase in daily insulin dose was more pronounced for males (19%) than for females (6%). Body mass index increased significantly in all treatment groups during the study period. The 1989 mean levels were higher in group B (males 20.6 kg m-2, females 22.0 kg m-2) than group A (males 19.3 kg m-2, p = 0.002; females 20.7 kg m-2, p = 0.000003) and group C (males 19.4 kg m-2, p = 0.0005; females 20.7 kg m-2, p = 0.008).(ABSTRACT TRUNCATED AT 250 WORDS)


Assessment of glucose turnover rates in euglycaemic clamp studies using primed-constant [3-3H]-glucose infusion and labelled or unlabelled glucose infusates.

Underestimation of glucose turnover rates has been a problem in clamp studies using primed-constant [3-3H]-glucose infusion technique. Due to slow mixing in interstitial compartments concealed specific activity gradients may arise between plasma and interstitial compartments during intravenous unlabelled glucose infusion. Such specific activity gradients, however, can be prevented if plasma specific activity is maintained constant. Two euglycaemic clamp studies (insulin infusion 40 mU m-2 min-1) were performed in six lean normal subjects. Using conventional unlabelled glucose infusates plasma specific activity declined by 74%, tracer determined glucose appearance was smaller than actual glucose infusion rates (317 +/- 11 vs 366 +/- 15 mg m-2 min-1, p < 0.001), and erroneous negative values were calculated for glucose production (- 49 +/- 7 mg m-2 min-1). Average underestimation during the first 2 h correlated with glucose infusion rates (r = 0.88, p < 0.02). In contrast, when plasma specific activity was maintained constant, using appropriately labelled glucose infusates, tracer determined glucose appearance and glucose infusion rates were similar (385 +/- 16 vs 385 +/- 17 mg m-2 min-1), and negative errors for glucose production were avoided. In conclusion, using unlabelled glucose infusates, as in previous studies, suppression of glucose production is overestimated and stimulation of glucose utilization is underestimated. As errors were greater with larger glucose infusions, the mistakes may have been greatest in insulin sensitive control subjects, and smaller in insulin resistant subjects. Therefore, re-evaluation of hepatic insulin sensitivity seems appropriate in diabetes, obesity, and other insulin resistant states.


Hospital admissions of adolescent patients with diabetes.

This study formed part of a psychological survey of young people aged 10-17 years attending three Bristol diabetes clinics. The aim was to examine the characteristics of those who were admitted to hospital with unstable diabetes during the period between two assessments, and analyse how they differed from the rest. Ten young people were admitted to hospital during the study period. These 10 individuals had greater emotional difficulty with diabetes as measured by diabetes specific psychometric scales (median scale score: 19) compared to the 89 adolescents who were not readmitted (median scale score: 14) (p = 0.01). They did not differ in demographic characteristics or glycaemic control. Overall there was no relationship between psychological response to diabetes and glycaemic control as assessed by mean glycated haemoglobin. Five of those admitted presented with hyperglycaemia; they had greater emotional difficulty (median scale score: 31, p = 0.02) and a more negative attitude (median scale score: 22, p = 0.02) to diabetes than those presenting with hypoglycaemia (median scale scores: 16). Only those with hyperglycaemia differed from those who were not readmitted, having greater emotional difficulty (p = 0.002) and a more negative attitude (p = 0.01). The possibility of psychological difficulties with diabetes should be sought following an admission, particularly for hyperglycaemia.


Benefits provided by an integrated education and clinical diabetes centre: a follow-up study.

The effects of a new integrated system of diabetes care with an enhanced role of the diabetes specialist nurse based in a purposed design diabetes centre, on diabetes control, attendance and cancellation rates, and admission for diabetic emergencies have been reviewed. Glycaemic control was examined in: (a) a cohort of 163 insulin-treated and 47 non-insulin treated diabetic subjects (age < 65 years) studied prospectively before and 3 years following the introduction of a new system of care; (b) a second cohort of more elderly patients aged greater than 65 years studied for the 3 years after the change over; (c) a cross-sectional study of the clinic population (n = 700) the year before and 3 years after the changeover; (d) a group of patients attending standard unaltered clinics in the same district (n = 157). Significant and sustained falls in HbA1 were observed in all groups of subjects attending the centre, with the means for those aged less than 65 falling from 11.9 +/- 2.3% to 9.9 +/- 1.9% and for those aged over 65 from a mean of 11.7 +/- 2.0% to 10.3 +/- 2.3%, 3 years later. The cross-sectional study provided similar results with a mean HbA1 of 12.2 +/- 3.0% prior to changeover and 10.4 +/- 4.4%, 3 years later. Smaller but significant changes were observed in patients continuing to attend the routine clinic (from 12.2 +/- 2.3% to 11.3 +/- 2.6%) over a similar period. Yearly admission rates for ketoacidosis and hypoglycaemia fell from 44 and 23, to 33 and 5 per annum, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


A community survey of diabetes in the elderly.

All 98 elderly patients who were known to have diabetes on the lists of two inner-city general practices were interviewed and examined in their own homes. They were characterized in terms of their prevalence of vascular complications, metabolic control, perceived health (measured by the Nottingham Health Profile), frequency of hospital and general practice contacts and use of domiciliary 'support services' and compared to 98 control subjects with an identical age/gender distribution. Mean haemoglobin A1 in the diabetic patients was 9.9 +/- 2.5%, few of them had suffered severe hyperglycaemia (10%) or hypoglycaemia (5%) and they were little more obese than controls (mean body mass index 26.2 +/- 4.2 vs 24.6 +/- 4.1 kg m-2; 95% confidence interval for the difference 0.4-2.8; p < 0.01). All complications were more prevalent in diabetic patients and the difference was statistically significant for visual impairment, strokes, impaired mental test scores, absent vibration sense, and absent leg pulses. Cataract was more common than retinopathy as a cause of visual disability. Diabetic patients perceived their health to be much worse than controls and were more often depressed. No measure of perceived health correlated with haemoglobin A1. We conclude that care of the diabetic elderly should not be too narrowly focused on 'metabolic' goals.


Diabetes self help facilitated by local diabetes research: the Coventry Asian Diabetes Support Group.

Diabetic South Asians know less about the nature of diabetes than Europeans. In view of the difficulties in communicating with diabetic South Asians in a clinical environment, a 'self help group' was established. The group was initially resourced by the Coventry Diabetes Study, became independent after 2 years and has met monthly for 4 years. Meetings are led and organized by local diabetic South Asians and include invited speakers and discussions. All meetings are held in one of the South Asian languages, particularly Punjabi. Attendance has ranged from 15 to 50, almost exclusively from the surveyed diabetic population of one electoral ward. Those with a high glycated haemoglobin (HbA1 > 9.5%) were assessed 12 months after invitation to the group, 44% (22/51) of whom attended at least twice. Those living over 1 mile from the meeting place and those speaking a minority language were least likely to attend. Those who attended the group had a greater drop in HbA1 and increase in 'knowledge score' than those not attending (both p < 0.01). The continued existence of the group illustrates the ability of local health workers to facilitate and empower local communities to become more responsible for their own health.


Thinking through uncertainty: nonconsequential reasoning and choice.

When thinking under uncertainty, people often do not consider appropriately each of the relevant branches of a decision tree, as required by consequentialism. As a result they sometimes violate Savage's sure-thing principle. In the Prisoner's Dilemma game, for example, many subjects compete when they know that the opponent has competed and when they know that the opponent has cooperated, but cooperate when they do not know the opponent's response. Newcomb's Problem and Wason's selection task are also interpreted as manifestations of nonconsequential decision making and reasoning. The causes and implications of such behavior, and the notion of quasi-magical thinking, are discussed.


Perceptual organization and attention.

It is widely assumed that the grouping of the visual field first described by the Gestalt psychologists and the related phenomenon of texture segregation occur very early in the processing of visual information and involve preattentive processes. All the recent evidence supporting this assumption comes from visual search experiments in which the subject is actively looking for a target and attending to the stimulus. The question at issue is whether these kinds of patterns are perceived under conditions of inattention, i.e., when observers are not searching for them. We performed six experiments to determine whether texture segregation and grouping by similarity or proximity are perceived under conditions of inattention. On the first two trials subjects were asked to report the longer arm of a briefly presented cross which was surrounded by a pattern of ungrouped small elements. On the third trial and subsequent control trials these elements were configured into grouping patterns and subjects queried about them immediately following their line length reports. The results establish that neither texture segregation nor grouping by similarity of lightness or proximity are perceived under conditions of inattention. They support the conclusion that there is an earlier stage of processing than that referred to as preattentive.


Perception without attention: results of a new method.

Having found by the use of a new method for examining perception without attention that grouping and texture segregation do not seem to occur (see Mack, Tang, Tuma, Kahn, & Rock (1992) Cognitive Psychology, 24, we go on to ask what is perceived without attention using this new method. Our subjects receive only one inattention trial in a sequence of trials involving a visual distraction task. In addition to the distraction task in the inattention trial, subjects received a stimulus of which they had no prior knowledge or expectation and were questioned or tested directly afterward for their perception of that stimulus. Two subsequent trials containing test stimuli serve as within-subject controls. The results of a series of experiments indicate that the presence of one or more stimulus objects and their locations are preattentively perceived, as is their color, but shape is not. Because individual items are detected without attention, we conclude that perceptual organization is initially based on a principle in which connected regions of uniform stimulation are inferred to be discrete units (the principle of uniform connectedness). One striking, unexpected finding is that without attention many subjects have no awareness at all of the stimulus object, an effect we call inattentional blindness.


A computer aided approach to electron energy loss spectroscopy and imaging.

In this paper we present the design and configuration of an imaging system for electron energy loss spectroscopy (EELS) and electron spectrographic imaging (ESI). The interfacing of commercial off-the-shelf hardware, custom software and a transmission electron microscope containing integrated spectrographic capabilities produces results that are well suited for clinical applications. The custom design and integration of this system allows for full control over all of the methods and procedures employed. This full control and knowledge of the procedures used is not always possible with commercial packages. This custom software allowed for the definition and testing of a new procedure for the determination of the theoretical background image, and hence the ability to refine the overall sensitivity for elemental detection. To further the proof of elemental detection, a procedure for continuous scan of an area of interest while varying delta E can also be performed with this system.


Does the heart electric activation split upon the infarcted area? An attempt at its detection by high-resolution electrocardiography.

Based on the presumption of the activation front splitting, the authors present their own proposal for the estimation of the myocardial electric activation course and for the detection of micropotentials 'hidden' within QRS complexes by the method of high-resolution electrocardiography. After filtration of QRS complex the values of delta RMS and those of cumulative amplitudes are calculated from the initial and from the terminal parts of QRS complexes. The presence of late potentials is reflected in a slowing down of the termination of activation course. As compared with healthy subjects, a slower rise of activation was observed in patients with myocardial infarction of the anterior wall. The curves of cumulative amplitudes rose very slowly during the first 70 ms of heart ventricle activation, explained according to the hypothesis of authors as being due to splitting of the activation front at the infarction focus. The usefulness of the proposed method was checked in patients with arterial hypertension and left ventricular hypertrophy, and in a group of patients with myocardial infarction. By the construction of cumulative amplitude curves from the onset of filtered QRS complexes, myocardial foci not reflected by 'classic' late potentials can be detected. The partial cumulative amplitudes of the QRS complex are suitable for comparative studies.


PKRD: a pharmacokinetic program for least-squares and bayesian analysis of repeated-dose pharmacokinetic curves.

A microcomputer program is presented which analyses multiple-dose pharmacokinetic curves using either a least-squares nonlinear analysis or a bayesian fit. The least-squares subroutine is designed to fit retrospective pharmacokinetic curves and can generate the so-called population pharmacokinetic parameters using the Standard Two-Stage method. The bayesian subroutine can instead be used prospectively to individualise the dosage regimen of a patient based on the concentrations measured in the initial phases of the drug treatment. All the pharmacokinetic subroutines of the program use the one-compartment model with either constant-rate intravenous infusion or oral route with zero-order absorption. The program was tested in a series of ten bone marrow transplantation patients treated with cyclosporine. The least-squares estimates of the one-compartment parameters, calculated by the program, were compared with those generated by the SIMKIN program.


NHPPREG: a computer program for the analysis of nonhomogeneous Poisson process data with covariates.

A program written in C is described which finds maximum likelihood estimates of regression coefficients and other relevant parameters for a point process with covariates. The data are assumed to come from a Poisson point process and may be modelled, at the user's choice, either totally parametrically or semi-parametrically. Furthermore, extra Poisson variation may be introduced, in both the parametric and semi-parametric approach, by the device of random effects. The work closely follows the theory developed by Lawless (1987).


Automatic visual field: a software diagnostic procedure.

A recently developed technique for the automatic acquisition of data on visual field losses was tested, with reliable neuro-ophthalmological parameters, on 300 subjects with and without eye disorders. Algorithms for acquisition and processing of the central visual field were implemented on an IBM-AT personal computer with standard peripherals. The contours of scotomata (visual field losses) were best estimated by probabilistic adaptive enhancement of data sampling in those areas with greater visual variability. Image processing tools were specially designed to extract information concerning the size, shape, position and number of scotomata in the visual field. The diagnosis of wider campimetric lesions required parameters such as symmetry or specularity coefficients, obtainable by analysing the visual field of both eyes. Data obtained were correlated to the pathology involved by univariate statistical tools.


A set of BASIC programs to evaluate relationships among protein sequences by optimum alignment and distance matrix analysis.

These six programs evaluate relationships among a number (> 2) of protein (or DNA) sequences. Program 1 automatically computes optimum alignments and total distances for all pairwise sequence combinations over any user desired range of the two gap penalties. Programs 2, 3 and 4 generate a square, symmetrical distance matrix, which can be exported for cluster analysis, or further analysed with programs 5 and 6 to give specific distances between sequences and extract sequence relationships. Data and results are exchanged among these programs, which are written in BASIC and compiled to run on Macintosh (68020/030/040) type machines with coprocessor and at least one MB of RAM. BASIC graphics commands or those for the Macintosh interface are avoided to facilitate use on other machines. Two groups of sequences are used to demonstrate (a) alignment, inter sequence distance calculation and dendrogram generation and (b) specific distance calculation and its usage in detecting sub groups of related sequences in dendrograms.


BB_RADABA: a computer program as a tool for breeding and management of laboratory animals.

The use of well characterized laboratory animals in biomedical research is of importance for quality securing and reproducibility of animal experiments. Therefore, breeders should not only have in mind the reproduction of their animal stocks, but they should also be interested to achieve an optimal resemblance of the animals in the main traits characterizing them as a model of human diseases. Specific software applications which helps the breeders in monitoring the main breeding parameters, selecting suitable breeding pairs and data processing are practically not available. This prompted us to develop a computer program for management of breeding and experimental data, which we called BB_RADABA. In this report we want to give a short survey on this computer program.


A Macintosh software package for simulation of human red blood cell metabolism.

We have developed a computer software package for Macintosh to simulate the metabolism and hemoglobin binding affinity of human red blood cell. The model is capable of simulating hemoglobin binding of ligands, metabolite concentrations, and metabolic fluxes at physiological steady state and in response to extracellular parameter variations, such as pH, osmolarity, glucose, and adenine concentrations. The kinetic parameters of enzymes, extracellular conditions, and initial intracellular metabolite concentrations can be specified by the user in order to model a particular situation. The software is use friendly, utilizing menu, window, and mouse to interact with the user. It also provides a pathway map of the red cell, which allows a direct access to enzyme kinetics by clicking the enzymes in the map.


APIS: a software for model identification, simulation and dosage regimen calculations in clinical and experimental pharmacokinetics.

APIS is a software package based on mathematical modelling which provides a reliable approach in optimizing drug therapy. It was designed to assist clinicians in interpreting blood drug levels so that drug therapy may be better and more cost-effective. It is a methodological approach to describe, predict and control the kinetic behaviour of a drug. This software incorporates the principle of Bayesian procedures, i.e. one can use all available patient information (population) to determine patient-specific parameter estimates. These estimates can then be used to design an optimal and individualized drug regimen. APIS is an attractive and useful tool for clinical and experimental pharmacokinetics. APIS may be used on any IBM compatible computer using the Microsoft-Windows environment. The software is menu driven to provide a very user-friendly tool for analysing pharmacokinetic data and for designing dosage regimens.