Immunohistochemical localization of delta sleep-inducing peptide-like immunoreactivity in the central nervous system and pituitary of the frog Rana ridibunda. The purpose of the present study was to investigate the distribution of delta sleep-inducing peptide in the brain and pituitary of the frog Rana ridibunda and to determine the possible effect of this nonapeptide on adrenocorticotropic hormone and corticosteroid secretion. Delta sleep-inducing peptide-like immunoreactive fibres were observed throughout the brain of the frog. These fibres generally exhibited the characteristics of glial cell processes. Scarce delta sleep-inducing peptide-positive fibres were seen in the olfactory bulb and in the periventricular areas of the telencephalon. In the diencephalon, numerous delta sleep-inducing peptide-containing processes were noted in the preoptic nucleus, the infundibular nuclei and the median eminence. A few cerebrospinal fluid-contacting cells were visualized in the ventral nucleus of the infundibulum. Delta sleep-inducing peptide-positive fibres were also observed in the mesencephalon, radiating through the different layers of the tectum. In the cerebellum, all Purkinje cells exhibited delta sleep-inducing peptide-like immunoreactivity. More caudally, numerous delta sleep-inducing peptide-positive fibres were noted in the vestibular nucleus of the rhombencephalon. A dense network of delta sleep-inducing peptide-containing fibres was seen in the pars nervosa of the pituitary. In the distal lobe, a population of endocrine cells located in the anteroventral region contained delta sleep-inducing peptide-immunoreactive material. Labelling of consecutive sections of the pituitary by delta sleep-inducing peptide and adrenocorticotropic hormone antiserum revealed that a delta sleep-inducing peptide-related peptide is expressed in corticotroph cells. The possible role of delta sleep-inducing peptide in the control of adrenocorticotropic hormone and corticosteroid release was studied in vitro, using the perifusion system technique. Administration of graded doses of delta sleep-inducing peptide (from 10(-8) to 10(-6) M) to perifused frog anterior pituitary cells did not affect the spontaneous release of adrenocorticotropic hormone. In addition, prolonged infusion of delta sleep-inducing peptide (10(-6) M) did not alter the stimulatory effect of corticotropin-releasing factor (10(-7) M) on adrenocorticotropic hormone secretion. Similarly, exposure of frog interrenal slices to delta sleep-inducing peptide did not induce any modification of spontaneous or adrenocorticotropic hormone-evoked secretion of corticosterone and aldosterone. Our results provide the first evidence for the presence of a delta sleep-inducing peptide-related peptide in lower vertebrates. The occurrence of delta sleep-inducing peptide-like immunoreactivity in specific areas of the brain suggests that the peptide may act as a neuromodulator.(ABSTRACT TRUNCATED AT 400 WORDS)


Signal transmission from pineal photoreceptors to luminosity-type ganglion cells in the lamprey, Lampetra japonica. In order to study the signal transmission from pineal photoreceptors to luminosity (achromatic)-type ganglion cells of the lamprey, Lampetra japonica, the electrical activity of these cell groups was investigated using intra- and extracellular electrodes. By intracellular recording, it was shown that the photoreceptor cells responded to flashes of light with hyperpolarizations, and the ganglion cells also hyperpolarized with concurrent suppression of spike discharges. Concerning the slow membrane potentials, the light intensity-response relationships of both cell groups followed the Naka-Rushton hyperbolic function. The intensity range over which the ganglion cells responded was broader than that of the photoreceptors. The spectral sensitivity curve of the luminosity-type ganglion cell coincided with that of the photoreceptor, showing a peak sensitivity at 525 nm. Membrane resistance of the ganglion cells increased during light stimulation. These results suggest that the luminosity-type ganglion cell receives and integrates signals from photoreceptors with various light sensitivities, having a peak spectral sensitivity at 525 nm. The synaptic mechanism from the photoreceptors to the ganglion cell is a type of disfacilitation.


Localization of GABAA-receptor gamma 2-subunit mRNA-containing neurons in the rat central nervous system. The localization of neurons containing mRNA of the gamma 2-subunit of GABAA receptors was examined in the rat central nervous system with in situ hybridization histochemistry using an oligonucleotide probe to the sequence of the gamma 2-subunit. Neurons containing the gamma 2-subunit mRNA were widely but unevenly distributed in the brain. The location of gamma 2-subunit mRNA-containing neurons differed from those containing alpha- or beta-subunits. According to our results brain regions can be divided into three categories: one containing only gamma 2-subunit, one containing mRNA of at least one subunit other than gamma 2 and one containing more than one other subunit but not the gamma 2-subunit. The distribution of strongly labeled nuclei partly coincided with that of glutamate decarboxylase, suggesting that the GABAA receptor gamma 2-subunit would be involved in an autoreceptive mechanism of the GABAergic transmission.


GABAB receptors in various in vitro and in vivo models of epilepsy: a study with the GABAB receptor blocker CGP 35348. The effect of the GABAB receptor blocker CGP 35348 on epileptic processes in vitro and in vivo was studied. In hippocampal slices of the rat maintained in vitro, CGP 35348 (100 microM) induced a moderate increase in the frequency of extracellularly recorded spontaneous epileptiform burst discharges induced in CA3 by penicillin (1.2 mM), bicuculline (5 microM) and low Mg(2+) (0.1 mM). This effect was observed in 50-75% of the slices. A similar but less consistent increase was also observed in CA1 in bicuculline and low Mg2+. Data obtained by intracellular recordings from CA1 pyramidal cells in the presence of bicuculline (10 microM) demonstrated that CGP 35348 (100 microM) increased the duration of the paroxysmal depolarization underlying an evoked epileptiform burst and reduced the early component of the after hyperpolarization which followed the burst. In mice pretreated with isoniazid, CGP 35348 (300 mg/kg, i.p.) significantly increased the number of convulsing mice. However, convulsions induced by submaximal doses of pentylenetetrazol, picrotoxin or strychnine were not facilitated by CGP 35348. We conclude that GABAB receptors appear to exert a suppressant effect on various kinds of epileptiform discharges of hippocampal neurons in vitro. In vivo, however, the role of GABAB receptors in regulating convulsions is less prominent since only isoniazid-induced convulsions were facilitated by GABAB receptor blockade.


Chronic reserpine administration selectively up-regulates beta 1- and alpha 1b-adrenergic receptors in rat brain: an autoradiographic study. Rats were treated for 15 days with reserpine or vehicle. One day after the last treatment, animals were killed and frozen brain sections were prepared for in vitro autoradiography. Binding to beta-adrenergic receptors was measured with [125I]iodocyanopindolol, and binding selective for beta 1 and beta 2 subtypes was assessed by including non-radioactive drugs that selectively mask beta receptor subtypes. Total alpha 1-adrenergic receptor binding was measured with [3H]prazosin, while alpha 1a binding was measured with [3H]WB4101 (in the presence of unlabeled serotonin). Quantitative densitometric analysis revealed that chronic reserpine treatment caused an increase in beta binding throughout the brain, including the cortex, thalamus, amygdala, hippocampus, caudate-putamen and hypothalamus. This effect of reserpine was entirely confined to the beta 1 subtype in all regions examined. [3H]Prazosin binding (alpha 1a plus alpha 1b) was also increased after chronic reserpine in several regions of the cortex and thalamus, as well as the ventral hippocampus and caudal amygdala. No effect of chronic reserpine was seen on [3H]WB4101 binding, indicating that the effect of reserpine on alpha 1 receptors is limited to the alpha 1b subtype. The increase in alpha 1b binding after reserpine administration in rats was generally smaller and less widespread than that seen with beta 1 binding. Thus the effect of reserpine upon noradrenergic neurotransmission demonstrates a high degree of receptor specificity and regional selectivity.


A highly selective kappa-opioid receptor agonist, CI-977, reduces excitatory synaptic potentials in the rat locus coeruleus in vitro. Intracellular recordings were made from neurons in a rat locus coeruleus slice preparation in vitro. A postsynaptic potential was evoked by electrical stimulation of afferents to the neurons. CI-977 ([5R-(5a,7a,8b)]-N-methyl-N-[7-(1-pyrrolidinyl)-1-oxaspiro[4.5]dec -8-yl[-4-benzofuranacetamide monohydrochloride) caused a depression of the evoked postsynaptic potential on locus coeruleus neurons. This action was reversed on washout. Bremazocine had a similar action on less than 50% of locus coeruleus neurons. Concentrations of CI-977 which depressed the postsynaptic potential did not affect either passive membrane conductance or a voltage-sensitive potassium current resembling IA. The depression of the excitatory postsynaptic potential caused by CI-977 remained in the presence of either 30 microM bicuculline and picrotoxin or when potassium acetate-filled recording electrodes were used. Using potassium chloride-filled recording electrodes and in the presence of 30 microM 6-cyano-2,3-dihydro-7-nitroquinoxaline-2,3-dione and either 30 microM DL-2-amino-5-phosphonovaleric acid or 500 microM kynurenic acid, CI-977 had no effect on the postsynaptic potential. The effects of CI-977 were reversed by 30-100 nM naloxone and 1-10 nM norbinaltorphimine but not by 1-10 nM naloxone. The hyperpolarizing response to the mu-opioid receptor-selective agonist D-Ala2,Nme Phe4,Gly-ol5 (DAGOL) was blocked by 1-10 nM naloxone but not by 1-100 nM norbinaltorphimine. The hyperpolarizing response to DAGOL was not affected by high doses of CI-977.(ABSTRACT TRUNCATED AT 250 WORDS)


Evidence of genetic heterogeneity among the nondystrophic myotonias. Recent in vitro electrophysiologic studies have demonstrated abnormal sodium channel gating in muscle from patients with Thomsen's disease and have called the chloride hypothesis into question. Abnormal sodium channel function, like myotonia, is a feature common to Thomsen's disease and several myotonias that are genetically linked to a chromosome-17q sodium channel locus. We present a pedigree segregating an allele for Thomsen's disease that is unlinked to this sodium channel locus, thus constituting evidence of genetic heterogeneity among the nondystrophic myotonias.


Nerve agents: a review.

Nerve agents: a review. Nerve agents produce neuromuscular blockade and convulsions in exposed humans. Military personnel in areas of potential exposure take prophylactic pyridostigmine. They are instructed to self-administer atropine and pralidoxime at the first sign of nerve agent toxicity. The key to treatment of nerve agent poisoning is the administration of atropine in doses larger than is customary in most other disorders, repeated as often as needed. Mechanical ventilation may be required. Convulsions are treated with diazepam, but only after atropine has been administered.


Synovial sarcoma. A clinicopathologic study. Twenty-five cases of synovial sarcoma were reviewed clinically, roentgenographically, and pathologically. The histopathology of each tumor was classified as either monophasic (19 cases) or biphasic (six cases). A soft-tissue mass was seen on roentgenography in 70% of patients; calcification was present in 15% of cases. Treatment included surgery, with or without radiation and/or chemotherapy. Fifteen patients remained disease free at an average of 7.5 years, one patient had pulmonary metastasis, and nine patients have died. The critical prognostic factor was adequate surgical margins; there were no local recurrences with adequate tumor-free margins, but there was an 83% recurrence rate with inadequate treatment.


Characteristics of Neisseria gonorrhoeae isolated in Australia showing decreased sensitivity to quinolone antibiotics. Forty three strains of Neisseria gonorrhoeae with decreased sensitivity to quinolone antibiotics were detected amongst 2141 Australian isolates of gonococci examined in the years 1984 to 1990. The strains examined belonged to 23 different auxotype/serovar classes, were generally more resistant to other antibiotics and, in the majority of cases, were isolated from travellers entering or returning to Australia from SE Asia. Quinolone-sensitive wild-type gonococci became less sensitive to these agents in vitro at a relatively high frequency when grown in the presence of quinolone concentrations at or around the MIC (Mean Inhibitory Concentration) of the antibiotic. Further increments in the levels of quinolone resistance of the already less-sensitive gonococci were also produced by this means, but high-level resistance to these agents was not observed. This suggests that mechanisms other than alterations in the DNA-gyrase of the organisms were responsible for the changes seen. Although spread of quinolone resistance in gonococci in Australia is unlikely to be rapid, if these antibiotics are used in therapy, treatment regimens with higher rather than lower dosages of quinolone antibiotics should be employed.


Bronchoscopy with bronchoalveolar lavage in the evaluation of pulmonary complications of bone marrow transplantation in children. Pulmonary complications are a frequent cause of morbidity and mortality following bone marrow transplantation. We examined the results of flexible bronchoscopy (FB) and bronchoalveolar lavage (BAL) in 27 pediatric bone marrow transplant (BMT) recipients with 29 episodes of pulmonary complications. Bone marrow transplant was performed for a variety of malignancies and hematologic disorders. Median age of BMT was 10.3 years (range, 1.7-17.6 years). Median time of FB following BMT was 60 days (range, 11-1,026 days). Routine cytologic and culture techniques were utilized to detect malignant cells, viruses, fungi, bacteria, and protozoa. Positive results were found in 15 (52%) with cytomegalovirus (CMV), the most common positive finding. In 14 (48%) episodes the results were negative. Fourteen patients had follow-up autopsy or open lung biopsy (OLB). Based on autopsy/OLB results, there were two false negatives and no false positives, giving a diagnostic sensitivity of 75% and specificity of 100%. There was one possible complication of FB and BAL. Survival of both positive and negative patients was poor, only seven patients being alive 90 days post-FB with BAL. We conclude that FB with BAL is a safe and accurate procedure for the diagnosis of pulmonary complications of BMT.


Human herpesvirus-6 DNA in cerebrospinal fluid of a child with exanthem subitum and meningoencephalitis. The involvement of human herpesvirus-6 (HHV-6) and herpes simplex virus infections was evaluated virologically and serologically in a 13-month-old girl with meningoencephalitic illness occurring in the pre-eruptive stage of exanthem subitum. An isolation of HHV-6 from blood and seroconversion to the virus confirmed the primary infection with the virus. HHV-6 gene sequences were detected in cerebrospinal fluid of acute stage of the disease by polymerase chain reaction. There was no evidence of herpes simplex virus infection in blood and cerebrospinal fluid. The patient recovered from the disease without any sequelae, although abnormal electroencephalography and cerebral computed tomography findings were observed temporally in the acute stage of the disease. These findings strongly suggest that HHV-6 invades the central nervous system and causes meningoencephalitis.


Relapse of herpes simplex encephalitis in children. The polymerase chain reaction method was used to diagnose herpes simplex encephalitis in children. Initial samples of cerebrospinal fluid from 15 patients with herpes simplex encephalitis were all positive for the herpes simplex virus DNA by polymerase chain reaction assay. In terms of early diagnosis, polymerase chain reaction assay became positive significantly earlier than the detection of intrathecally produced anti-herpes simplex virus antibody using the enzyme-linked immunosorbent assay (4.4 vs 8.9 days after onset; P less than .01). Serial examinations showed that the presence of virus DNA in cerebrospinal fluid continued for 3 to 18 days after the neurologic onset (mean 10.1 days). Four of the 15 patients had a relapse of encephalitis after completing acyclovir therapy. The mean duration of initial acyclovir therapy in the recurrent group was significantly shorter than that in the nonrecurrent group. In recurring cases, herpes simplex virus DNA reappeared temporarily in the cerebrospinal fluid of two patients. These results show that polymerase chain reaction assay is a useful diagnostic tool for the early and noninvasive diagnosis of herpes simplex encephalitis in children. Results also suggest that a comparatively short duration of acyclovir therapy may be related to a relapse of herpes simplex encephalitis in some children.


[2D time of flight stereoscopic MR angiography of pulmonary vessels]. Two-dimensional (2D) time-of-flight (TOF) stereoscopic MR angiographies (MRA) of the pulmonary vessels were obtained from 15 healthy volunteers and five patients with pulmonary cancer in the mediastinum and pulmonary hilum. Fifteen healthy volunteers were examined using FLASH (Fast Low Angle Shot) with breath holding (40/8/40, TR/TE/flip angle). Except for the left superior pulmonary vein, pulmonary vessels in the mediastinum and hilum were well defined on stereoscopic MRA images. Although it was difficult to define the pulmonary arteries in the peripheral zone, intersegmental veins were easily defined with this method. In five cases of pulmonary cancer that were confirmed to show definite tumor involvement of the pulmonary vessels in the mediastinum and hilum by enhanced CT and MRI (SE method), irregular narrowing and interruption of the vessels were shown on MRA. In conclusion, 2D TOF stereoscopic MRA is considered a noninvasive, effective method for evaluation of the morphology of pulmonary vessels adjacent to the tumor in the mediastinum and hilum.


[Doppler sonography in the diagnosis of liver tumors]. The value of color duplex Doppler sonography in evaluating tumor vascularity was investigated in 82 hepatic tumors (61 hepatocellular carcinomas, 11 metastatic cancers, eight adenomatous hyperplasia, one focal nodular hyperplasia, and one cholangiocellular carcinoma) receiving angiography, 64 intrahepatic arteries, and five hepatic cysts. The minimum diameter of the intrahepatic arteries (lateral inferior subsegmental arteries) from which signals could be weakly obtained by using a 3.5 MHz transducer was 0.7 mm. Twenty-eight (74%) of 38 tumors with signals within them had definite tumor vessels on angiography, and continuous blood flow within the tumors showed an association with the dilated tumor vessels. Eighteen (69%) of 26 tumors with signals within them receiving conventional angiography had tumor vessels greater than 0.7 mm. However, only 17 (31%) of 55 tumors less than or equal to 3 cm showed signals within them in contrast to 21 (78%) of 27 tumors greater than 3 cm. Three of eight adenomatous hyperplasias, which were angiographically undetected and had portal or hepatic venous branches, showed signals within them. Four tumors that had abnormally high velocity arterial signals (greater than 0.63 m/sec) within them showed no arteriovenous shunt. Evaluation of tumor vascularity according to the Doppler sonographic findings at the periphery of the tumor was difficult. This was attributed to the fact that the real sample volume was larger than that on B-mode image, with no correlation seen between the signals at the tumor periphery or the existence of arteries surrounding the tumor and tumor vascularity. Although a correlation was seen between tumor vascularity or tumor size and peak systolic velocity determined at the tumor periphery (p less than 0.05), five of six tumors with abnormally high velocities (greater than 0.63 m/sec) at the tumor periphery were greater than or equal to 5 cm in diameter. Doppler signals of the artery feeding the arteriovenous shunt were characterized by abnormally high velocity and low resistive index. In conclusion, Doppler sonography is somewhat useful in evaluating tumor vascularity, but less so in small hepatic tumors.


[Clinical studies for usefulness of Gd-DTPA enhanced MRI in lung cancer]. To evaluate utility of Gd-DTPA enhanced MRI (Gd-MRI) in lung cancer, Gd-MRI was performed in 69 cases. 1) Viable tumor was strongly enhanced, necrosis in the tumor, however, was not enhanced on Gd-MRI. Enhanced patterns of Gd-MRI were divided into 3 types, however there was little correlation between the enhancement patterns and histologic types. 2) In serial scan studies of 15 cases, the signal intensity of the tumor reached the peak 3 minutes to 10 minutes after Gd-DTPA administration, and after that the signal intensity decreased gradually. 3) In 23 of 27 (85%) hilar lung cancer cases, Gd-MRI could differentiate the tumor from the peripheral obstructive pneumonia or atelectasis. In 18 of these 23 cases, the peripheral lung disease showed higher intensity than the tumor. 4) In Gd-MRI of pulmonary nodules less than 3 cm in diameter, lung cancers (n = 13) were more strongly enhanced than tuberculomas (n = 5) (p less than 0.001). Based on these data, Gd-MRI was helpful for detecting tumor necrosis and tumor extension on hilar lung cancer with peripheral lung disease. Moreover Gd-MRI may become a feasible diagnostic method for pulmonary nodules.


[Evaluation of breath-hold multislice dynamic MRI of hepatocellular carcinomas]. The breath-hold multislice dynamic study (BMDS) in MRI, which can scan the entire liver during a single breath-holding, was applied to 16 patients with 30 focal lesions of hepatocellular carcinoma (HCC). The BMDS was performed at 20 seconds and 3 minutes after the bolus injection of Gd-DTPA, by gradient echo pulse sequence (FLASH). 29 nodules were detected in the BMDS, showing rapid enhancement on early phase and decline on delayed phase images. The BMDS was more sensitive than conventional MR images. Therefore, the BMDS seems to be useful for the diagnosis of HCC with multiple as well as solitary nodules.


Random mutagenesis of Schizosaccharomyces pombe SRP RNA: lethal and conditional lesions cluster in presumptive protein binding sites. Signal recognition particle (SRP), a ribonucleoprotein composed of six polypeptides and one RNA subunit, serves as an adaptor between the cytoplasmic protein synthetic machinery and the translocation apparatus of the endoplasmic reticulum. To begin constructing a functional map of the 7SL RNA component of SRP, we extensively mutagenized the Schizosaccharomyces pombe SRP7 gene. Phenotypes are reported for fifty-two mutant alleles derived from random point mutagenesis, seven alleles created by site-directed mutagenesis to introduce restriction sites into the SRP7 gene, nine alleles designed to pinpoint conditional lesions, and three alleles with extra nucleotides inserted at position 84. Our data indicate that virtually all single nucleotide changes as well as many multiple substitutions in this highly structured RNA are phenotypically silent. Six lethal alleles and eleven which result in sensitivity to the combination of high temperature and elevated osmotic strength were identified. These mutations cluster in conserved regions which, in the mammalian RNA, are protected from nucleolytic agents by SRP proteins. The effects of mutations in the presumptive binding site for a fission yeast SRP 9/14 homolog indicate that both the identity of a conserved residue and the secondary structure within which it is embedded are functionally important. The phenotypes of mutations in Domain IV suggest particular residues as base-specific contacts for the fission yeast SRP54 protein. A single allele which confers temperature-sensitivity in the absence of osmotic perturbants was identified in this study; the growth properties of the mutant strain suggest that the encoded RNA is somewhat defective even at the permissive temperature, and is most likely unable to correctly assemble with SRP proteins at the nonpermissive temperature.


Unusual features of the retroid element PAT from the nematode Panagrellus redivivus. The PAT retroid transposable elements differ from other retroids in that they have a 'split direct repeat' structure, i.e., and internal 300bp sequence is found repeated, about one half at each element extremity. A very abundant transcript of about 900 nt, the start of which maps to the preferentially deleted portion of PAT elements, is detected on total Panagrellus redivius RNA bearing Northern blots. A potentially corresponding ORF encodes a protein of 265 residues having a carboxy terminal Cystein motif, believed to be exclusively characteristic of the GAG protein in retoid elements. A much fainter, 1800nt long transcript, is also detected on Northern blots and maps slightly downstream of the first ORF. The predicted protein sequence of this region bears motifs typical of reverse transcriptase and RNaseH, as found in the Pol genes of retroid elements. Peptide motif similarities are greatest with the DIRS-1 element derived from Dictyostelium discoideum. The possibility of using PAT elements as transposon tagging system for Caenorhabditis elegans is discussed.


Towards identification of cis-acting elements involved in the replication of enterovirus and rhinovirus RNAs: a proposal for the existence of tRNA-like terminal structures. On the basis of a comparative analysis of published sequences, models for the secondary structure of the 3'-terminal [poly(A)-preceding] untranslated region of the entero- and rhinovirus RNAs were worked out. The models for all these viruses share a common core element, but there are an extra enterovirus-specific element and still an additional element characteristic of a subset of enterovirus RNAs. The two latter models were verified for poliovirus and coxsackievirus B genomes by testing with single-strand and double-strand specific enzymatic and chemical probes. A tRNA-like tertiary structure model for the 3'-terminal folding of enterovirus RNAs was proposed. A similar folding was proposed for the 3' termini of the negative RNA strands as well as for the 5' termini of the positive strand of all entero- and rhinovirus RNAs. Implications of these data for template recognition during negative and positive RNA strands synthesis and for the evolution of the picornavirus genomes are discussed.


Modifications of the E.coli Lac repressor for expression in eukaryotic cells: effects of nuclear signal sequences on protein activity and nuclear accumulation. Eukaryotic expression vectors designed to produce E. coli Lac repressor protein targeted to the nucleus of mammalian cells were constructed. These constructions carry the lac repressor gene (lacI) fused at different positions to a nuclear localization sequence (NLS) from either the SV40 large T antigen or the adenovirus E1a. When the NLS's were fused to the lacI gene at the 5' end, the protein produced exhibited tighter repression of beta-galactosidase expression than the unmodified LacI protein. Localization sequences at the extreme 3' end of the gene generally diminished induction by IPTG, while introduction of the SV40 NLS nine base pairs upstream of the 3' end eliminated repressor activity. When either NLS was placed at the 3' end behind a random nine base pair linker, the activity of the LacI protein depended on the sequence of the linker, and in 9 of 10 linkers tested, activity of the protein was adversely affected. The one exception was the fusion protein from p3'ss, which had the NLS at the 3' end of lacI behind the nine base pair linker, AGC AGC CTG (ser-ser-leu). This protein exhibited efficient nuclear accumulation, strong repressor activity and greater sensitivity to IPTG induction. The functional linker from the p3'ss fusion protein extends the leucine zipper heptad repeat located at the C-terminus of the protein. These data support the role of the leucine zipper in tetramer formation and predict that extension of this zipper will further stabilize the protein. This modified lacI gene should be valuable for improved adaptation of the prokaryotic regulatory system to eukaryotic cells.


Cis-regulation of the L-type pyruvate kinase gene promoter by glucose, insulin and cyclic AMP. The glucose/insulin response element of the L-pyruvate kinase gene is a perfect palindrome located from nt -168 to -144 with respect to the cap site. This element (L4) is partially homologous to MLTF binding sites. Its full efficiency requires cooperation with a contiguous binding site for HNF4, termed L3 and located from nt -145 to -125. In the presence of the L4 element contiguous to L3, cyclic AMP inhibits activity of the L-PK promoter while in its absence, or when the normal L4-L3 contiguity is modified, cyclic AMP behaves as a transcriptional activator that does not seem to be sequence-specific. Therefore, we propose that the mechanism of inhibition of the L-PK gene by cyclic AMP requires precise interactions between the nucleoprotein complex built up at sites L4 and L3 and other components of the L-PK transcription initiation complex.


Eukaryotic topoisomerase II cleavage of parallel stranded DNA tetraplexes. A guanine-rich single-stranded DNA from the human immunoglobulin switch region was shown by Sen and Gilbert [Nature, (1988) 334, 364-366] to be able to self-associate to form a stable four-stranded parallel DNA structure. Topoisomerase II did not cleave the single-stranded DNA molecule. Surprisingly, the enzyme did cleave the same DNA sequence when it was annealed into the four-stranded structure. The two cleavage sites observed were the same as those found when this DNA molecule was paired with a complementary molecule to create a normal B-DNA duplex. These cleavages were shown to be protein-linked and reversible by the addition of salt, suggesting a normal topoisomerase II reaction mechanism. In addition, an eight-stranded DNA molecule created by the association of a complementary oligonucleotide with the four-stranded structure was also cleaved by topoisomerase II despite being resistant to restriction endonuclease digestion. These results suggest that a single strand of DNA may possess the sequence information to direct topoisomerase II to a binding site, but the site must be base paired in a proper manner to do so. This demonstration of the ability of a four-stranded DNA molecule to be a substrate for an enzyme further suggests that these DNA structures may be present in cells.


The effects of Escherichia coli heat-stable enterotoxin in renal sodium tubular transport. To compare the function of sodium transport between intestine and renal tubule, we studied the effect of E. coli STa enterotoxin and 8-bromo cyclic GMP in perfused rat kidneys. Infusion of STa enterotoxin (0.017 and 0.1 micrograms/ml) caused a dose and time dependent decrease in total renal sodium tubular transport. The major site of STa effect was at the renal proximal tubule. Similar to Sta enterotoxin, 8-bromo cyclic GMP (10(-5) M) caused a significant decrease of fractional renal sodium proximal tubule transport. In contrast to STa enterotoxin, infusion of 8-bromo cyclic GMP resulted in a significant but short lasting (30 min.) increase in glomerular filtration rate. STa enterotoxin also decreased significantly the renal tissue potassium. This STa effect was related to a significant decrease in renal potassium tubular transport, resulting also in an increase of urinary potassium excretion. These studies demonstrate the specific functional effect of STa enterotoxin in promoting the decrease in renal proximal tubular sodium transport, similar to 8-bromo cyclic GMP. In perfused rat kidneys STa also decreased tissue potassium, mainly by a decrease in potassium transport and increase in urinary potassium excretion. These effects suggest the existence of an endogenous peptide resembling STa enterotoxin, that regulates the function of renal sodium tubular transport.


[Myeloperoxidase, lactoferrin and elastase in bronchoalveolar lavage and plasma in pneumonia]. Neutrophilic granulocytes in the lower respiratory tract are of decisive importance for the elimination of pathogenic germs in bacterial pneumonia. On the other hand, the liberation of phagocyte products (e.g. elastase) can result in tissue damage in the parenchyma of the lungs. For this reason, we determined in patients suffering from acute pneumonia (n = 21), in patients with acute pneumonia associated with immunosuppression (n = 12), in patients who had overcome their pneumonia (n = 9) and in controls (n = 17) in bronchoalveolar lavage (BALF) and in plasma, the concentration of the locally produced granulocyte products myeloperoxidase (MPO), lactoferrin (LF) and elastase-alpha 1 proteinase complex (ELA) as well as of the alpha 1 proteinase inhibitor (alpha 1 Pi) and alpha 2 proteinase inhibitor (alpha 2 Pi) via chemoluminescence immunoassay, and compared the same with the differential cell count in the BALF. The protein concentrations were referred to the albumin concentration (Alb) for standardisation. This concentration did not differ significantly between the various patients and control groups. The BALF concentration of ELA in the group with pneumonia (median: 86.3 micrograms/l or 8.5 micrograms/mg Alb) was about eight times higher than in the group of patients suffering from pneumonia with immunosuppression (median: 16 micrograms/l or 1.0 micrograms/l Alb, p less than 0.001) or in whom the pneumonia was no longer present (17.6 micrograms/l or 0.5 micrograms/mg), and approximately 40 times higher than in the control group (3 micrograms/l or 0.2 micrograms/mg, respectively). Similar results were obtained for LF (61 micrograms/mg Alb vs. 11.3; 16.8 and 5.9 micrograms/mg; p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


Primary mutations in calmodulin prevent activation of the Ca(++)-dependent Na+ channel in Paramecium. Paramecium tetraurelia behavioral mutant cam12 displays a "fast-2" behavioral phenotype: it fails to respond to Na+ stimuli. Electrophysiologically, it lacks a Ca(++)-dependent Na+ current. Genetics and DNA sequencing showed the primary defect of cam12 to be in the calmodulin gene (Kink et al., 1990). To correlate calmodulin structure and function in Paramecium, we elucidated the primary structure of cam12 calmodulin. Peptide sequencing confirmed the two point mutations predicted by the DNA sequence: a glycine-to-glutamate substitution at position 40 and an aspartate-to-asparagine substitution at position 50. Our results further showed that lysine 13 and lysine 115 were methylated normally in cam12. It is likely that the electrophysiological abnormalities of cam12 are a direct reflection of the amino-acid substitutions, as opposed to improper posttranslational modification.


Intravenous infusion of n-3 polyunsaturated fatty acids. Dietary supplementation with n-3 polyunsaturated fatty acids (PUFA) is regarded as beneficial for the prevention and treatment of atherosclerosis and thrombosis and chronic inflammatory diseases like rheumatoid arthritis and psoriasis. It may be possible to treat some acute diseases like acute myocardial infarction or acute rejection of grafted organs if it is possible to make n-3 PUFA take effect quickly (in hours instead of days). Three sets of experiments were done. In Experiment 1, emulsion of trieicosapentaenoyl-glycerol (EPA-TG) and tridocosahexaenoyl-glycerol was infused through rabbit ear veins, and the leukotriene B4/B5 production from polymorphonuclear leukocytes was measured at different time points by high-performance liquid chromatography. In Experiment 2, delayed type hypersensitivity (DTH) of mice was measured with sheep red blood cells as an antigen. Pure n-3 PUFA emulsions or a control solution were infused through tail veins just before the second challenge of the antigen. DTH was measured 24 hr after the second challenge. In Experiment 3, human natural killer cell activity was measured using K562 target cells before and after the infusion of pure EPA-TG emulsion to an antecubital vein. Leukotriene B4 production by rabbit polymorphonuclear leukocytes was depressed by 40% by EPA-TG infusion. DTH was suppressed almost completely by n-3 PUFA infusion. Natural killer cell activity was suppressed almost completely by EPA-TG infusion in 8 hr. DTH, natural killer cell activity, and leukotriene B4 production are probably related to acute rejection of grafted organs.(ABSTRACT TRUNCATED AT 250 WORDS)


[Germ cell tumors in children. Evaluation of regional cases over a 23 year period]. A retrospective study on germ cell tumors on children is reported. The purpose of this study was to evaluate some prognostic factors (age, sex, site, histology, treatment) as well as to compare the casistic of Puglia Region with the results of the National literature. 45 patients (30 females, 15 males) aging 2 days-12 years observed in three Pediatric Centres of Bari into 23 years. Primary site of the tumor was: ovary 9, testis 9, sacrococcygeal 16, and others site 11 (head 4, mediastinum 2, pelvis 2, abdomen 2, gluteus 1). teratoma 31, choriocarcinoma 1, embryonal carcinoma 7, endodermal sinus tumor 5, germinoma 1. surgery 35 patients, surgery+chemotherapy 6, surgery+chemotherapy+radiotherapy 4. Results of treatment: 37 of 41 valuable patients achieved a response (34 complete, 3 partial response); 4 patients had no response. Of the 37 patients that have had a complete or partial remission, 8 subsequently relapsed; of these 8 patients, 5 obtained a second remission after second line therapy, 3 are dead for progressive disease. Better prognosis was observed in children with gonadal tumors treated with surgery alone.


Antioxidants in health and disease: overview. Molecular oxygen is an essential nutrient for higher forms of life. In addition to its normal physiological reactions, oxygen and its partially reduced forms can oxidize a variety of macromolecular and simpler compounds in cells and fluids of the body. Such oxidized and peroxidized compounds have been associated with, and may be causally related to, a variety of chronic diseases. As a protection against excessive oxidation, nature has developed a complex set of interactive antioxidant systems. Selected aspects of antioxidant actions are considered in this symposium.


Clinical uses and abuses of vitamin E in children. The major benefits arising from elevated dosages of vitamin E have been the relief of symptoms of vitamin E deficiency in humans with abetalipoproteinemia and chronic cholestasis. In addition, supplements of vitamin E prevent the isolated vitamin E deficiency that has recently been associated with spinocerebellar symptoms. In keeping with the view that newborn infants, and especially premature infants, suffer from vitamin E deficiency, elevated dosages of vitamin E have been administered to prevent the anemia of premature infants, retrolental fibroplasia, bronchopulmonary dysplasia, and intraventricular hemorrhage. However, the results have been conflicting. Furthermore, some infants treated with vitamin E die unexpectedly. The life-threatening hazard of such treatments has been attributed mainly to polysorbates that are used as detergents in preparations of vitamin E for intravenous use rather than to vitamin E itself. The possibility that vitamin E, in its action as an antioxidant, inhibits the generation of superoxide anion in leukocytes is examined in this paper.


Activation of a single alpha-1-adrenoceptor subtype in rat aorta mobilizes intracellular and extracellular pools of calcium. The calcium channel antagonist, nifedipine (0.1 microM), only slightly inhibited the response to the full alpha 1-adrenoceptor agonist, (-)-norepinephrine, in rat aorta, but nearly completely inhibited the response to the partial alpha 1-adrenoceptor agonist, (-)-dobutamine, indicating that (-)-norepinephrine primarily utilizes intracellular stores of calcium to produce vasoconstriction, whereas (-)-dobutamine relies primarily upon the translocation of extracellular calcium. It has been proposed that the different pools of calcium mobilized by (-)-norepinephrine and (-)-dobutamine in rat aorta result from the activation of two different alpha 1-adrenoceptor subtypes by these agonists. Because the irreversible alpha 1-adrenoceptor antagonist, phenoxybenzamine, has been proposed to inactivate an alpha 1-adrenoceptor subtype coupled specifically to the mobilization of intracellular stores of calcium, this compound should selectively inhibit the response of (-)-norepinephrine, and not the response of (-)-dobutamine, if two distinct alpha 1-adrenoceptor subtypes exist in rat aorta. In the present study, phenoxybenzamine produced concentration-dependent, noncompetitive inhibition of the responses to both agonists in rat aorta, and the pA2' values for phenoxybenzamine were 8.12 +/- 0.23 and 7.74 +/- 0.27 against (-)-dobutamine and (-)-norepinephrine, respectively, which are not significantly different from each other (p greater than 0.05). In addition, the phasic and tonic components of the response to (-)-norepinephrine, which are mediated by the mobilization of intracellular calcium and the translocation of extracellular calcium, respectively, are also inhibited to the same extent by phenoxybenzamine. The results do not support the hypothesis of two alpha 1-adrenoceptor subtypes in rat aorta, one of which being coupled to the translocation of extracellular calcium, and the other to the mobilization of intracellular calcium. Rather, the results support our previous hypothesis that a single alpha 1-adrenoceptor subtype exists in vascular smooth muscle, and this alpha 1-adrenoceptor is coupled to two distinct signal transduction processes, one of which elicits the mobilization of intracellular calcium, and the other opens membrane calcium channels to permit the influx of extracellular calcium.


The kappa-opioid U-50,488H suppresses the initiation of nocturnal spontaneous drinking in normally hydrated rats. The effect of a systemic (IP) treatment with 1.0, 3.0 and 9.0 mg/kg U-50,488H (U50), a highly selective kappa-agonist, on spontaneous, nocturnal ingestive behavior of the rat was studied using a microcomputer controlled data acquisition system. The latency to initiate drinking was increased and drinking behavior was suppressed in the first hour after injection in a dose-dependent manner. The consummatory indices of drinking were not affected. After this period of adipsia, a phase of polydipsia, that was probably due to the diuretic effect of U50, was evident. This prophagic effect of U50 was evident only at the dose of 3 mg/kg and was accompanied by an increased duration of feeding episodes but not by a reduced latency to feed. These results suggest that kappa-receptors play a pivotal role in modulating spontaneous drinking in the normally hydrated rat and that this control is mainly exerted on the motivational aspect of drinking.


Social isolation does not alter brain regional benzodiazepine binding site numbers, affinity and coupling in the rat. Male Sprague-Dawley rats were differentially housed for 21 days immediately after weaning. Isolated animals showed a selective suppression of exploration of the light side of a two compartment box; spending significantly less time in the light, and making fewer transitions between the light and dark compartments compared to socially reared controls. However, both basal and GABA-stimulated [3H] flunitrazepam binding was unaltered in the frontal cortex, hippocampus, amygdala and cerebellum following social isolation. These results are discussed in relation to other studies on central benzodiazepine receptor changes following a variety of experimental stressors.


Diazepam antagonizes effects on dopamine metabolism produced by PCP receptor agonists. 1. Male rats were injected with either saline, diazepam, MK-801, or diazepam plus MK-801. 2. In previous work with phencyclidine (PCP), diazepam significantly reduced the increase in homovanillic acid (HVA) in olfactory tubercle and prefrontal cortex. 3. Diazepam also lowered the HVA increase following MK-801 in caudate, olfactory tubercle, and prefrontal cortex. 4. Benzodiazepine receptors may modify dopaminergic function at PCP receptors that affect dopamine neurotransmission.


Rat gastric relaxation induced by stimulation of endothelin-1 selective receptors. We have investigated the smooth muscle activity of ET-1 and ET-3 on rat fundus strips in vitro as well as the effects of the peptides on gastric motility in vivo. In the isolated tissue with no precontraction ET-1 and ET-3 were potent spasmogens which produced half maximal contractions at concentrations 4.5 and 8.0 nM, respectively. In contrast, under conditions where the isolated tissue was precontracted to approx. 50% of maximum by prostaglandin E2, ET-1 dose-dependently (5 x 10(-10) - 10(-8) M) and temporarily relaxed the fundus strip, whereas ET-3 further increased the contraction. The relaxing capacity of ET-1 was absent when the tissue was precontracted by potassium yet was resistant to pretreatments with tetrodotoxin, capsaicin, propranolol, indomethacin, NG-methyl-L-arginine or glibenclamide. In addition in vivo ET-1 and ET-3 (less than 1 nmol/kg) showed opposite effects on gastric motility as the former reduced basal tonus and spontaneous activity, whereas the latter increased the motor activity of the gastric ventricle. The results support the notion that ET-1 may induce gastric relaxation by stimulation of selective receptors whereas stimulation of nonselective receptors may promote gastric smooth muscle contraction.


Cysteamine induced-duodenal ulcers are associated with a selective depletion in gastric and duodenal calcitonin gene-related peptide-like immunoreactivity in rats. We have measured the endogenous levels of gastric and duodenal calcitonin gene-related peptide (CGRP)-, neurokinin A (NKA)-, galanin-vasoactive intestinal polypeptide (VIP)- and neuropeptide Y (NPY)-like immunoreactivity (li) in relation to cysteamine-induced gastric lesions and duodenal ulcers in rats. CGRP-li but not NKA-, galanin-, VIP- or NPY-li was decreased in gastric and duodenal samples following a single ulcerogenic dose of cysteamine (900 mg/kg p.o.). Temporal relationships of this phenomenon showed that CGRP-li was selectively decreased (stomach 45%, duodenum 68% as compared to controls, respectively after 24 h) concomitantly to the formation of acute gastric lesions and duodenal ulcers. Animals bearing healed ulcers 12 days after cysteamine, had gastroduodenal CGRP-li similar to control values. Pretreatment with the selective sensory neurotoxin capsaicin decreased gastroduodenal CGRP-li but not NKA-, galanin-, VIP- or NPY-li, showing that CGRP might be considered a marker of the afferent innervation of the gastroduodenal tract. The residual gastroduodenal CGRP-li levels in capsaicin-pretreated animals were not decreased by cysteamine administration, indicating that the effect of cysteamine is restricted to a peptide pool of primary afferent origin. Duodenal CGRP-li is selectively decreased by the duodenal ulcerogen cysteamine during the acute phase of ulcers formation and might be among the local mediators which afford protection against the ulcerogenic stimuli.


[ACTH-independent Cushing's syndrome due to bilateral macronodular adrenal hyperplasia with empty sella turcica and anterior panhypopituitarism]. A case is presented of Cushing's syndrome due to macronodular bilateral adrenal hyperplasia which is ACTH-independent as was demonstrated by the undetectable basal and after stimulation with metoprolol ACTH plasma levels. High cortisol levels is associated in this patient with empty sella turcica and anterior panhypopituitarism with confirm the exclusive adrenal origin of the hormone hypersecretion and the lack of treatment success with hypophysis ablation in this process.


Multiple organ failure with the adult respiratory distress syndrome in homicidal arsenic poisoning. A 30-year-old man and a 39-year-old woman, who was 28 weeks pregnant, were simultaneously poisoned by eating chocolate containing arsenic trioxide. They developed a picture of multiple organ failure peaking around the 8th to 10th day after ingestion, with the development of life-threatening adult respiratory distress syndrome (ARDS) in both patients. This rarely reported complication of arsenic poisoning was managed successfully by intubation and mechanical ventilation with positive end expiratory pressure in both patients. Hemodynamic and laboratory data are presented supporting the clinical course. Arsenic toxicity further resulted in intrauterine fetal death. The effects of severe arsenic poisoning leading to early multiple organ failure with ARDS as well as to protracted, debilitating polyneuropathy are discussed.


[Use of a local antibiotic combination in the treatment of mycotic vaginitis and mixed vaginitis]. This open clinical trial in 121 patients carried out by gynaecologists shows the difficulty in establishing the clinical diagnosis of vaginal candidosis or vaginal infectious candidosis. On the other hand, specific vaginitis can easily be identified, without carrying out vaginal smear. Bacteriological and clinical arguments show the efficacy of using an antibiotic association by the vaginal route for the treatment of these types of vaginitis (candidosis or infectious candidosis). The product is particularly interesting in all cases where vaginal smear have not been taken.


IgG is bound by antigen-antibody bonds and some IgG and albumin are bound by intermolecular disulfide bonds to cartilage in rheumatoid arthritis and osteoarthritis. To elucidate the mechanisms for the presence of immunoglobulins and human serum albumin (HSA) in articular cartilage from patients with rheumatoid arthritis (RA) or osteoarthritis (OA), the recovery of these molecules was determined in several elution steps. These steps included serial elutions with a neutral buffer to extract entrapped molecules, elution with 6 M guanidine hydrochloride to extract molecules bound by noncovalent interactions, and digestion of cartilage with bacterial collagenase to release molecules covalently bound to cartilage matrix proteins. Significantly more IgG than HSA was recovered with 6 M guanidine after serial elutions with neutral buffer from the cartilages of patients with both RA and OA, consistent with the binding of IgG by antigen-antibody bonds. Degradation of cartilage with collagenase released additional IgG and HSA. Analysis of the IgG and HSA, recovered with guanidine or with collagenase, using SDS-PAGE and transfer blotting, indicated for the first time the presence of disulfide bonds between these molecules and cartilage matrix molecules.


B lymphoblasts show oxidase activity in response to cross-linking of surface IgM and HLA-DR. Human B lymphocytes express components of the superoxide generating system of phagocytes, NADPH oxidase. We studied regulation of this 'B-cell oxidase' during in vitro blast transformation, using Lucigenin-amplified chemiluminescence (CL) to detect superoxide release. While freshly isolated tonsil B lymphocytes showed no CL responses, culture with phorbol myristate acetate (PMA) and ionomycin induced susceptibility to CL triggering by anti-IgM and anti-HLA-DR. Maximal effects were observed after 3 days of culture with 0.4 ng/ml PMA + 1 microgram/ml ionomycin. Cells from such B lymphoblast cultures showed no CL responses to opsonized zymosan. In contrast, peripheral blood mononuclear cells, where monocytes are the predominant oxidant source, showed CL responses to opsonized zymosan but not to anti-IgM and anti-HLA-DR, either before or after culture with PMA + ionomycin. Culture of B cells with the surface immunoglobulin cross-linking agent staphylococcus aureus Cowan I also led to emergence of a CL, response to anti-IgM, which was enhanced by interferon-gamma. Interestingly, markedly fewer B blasts than freshly isolated B lymphocytes expressed cytochrome b-558 surface antigen. Thus, the B-cell oxidase is up-regulated during blast transformation and can be triggered via surface IgM and HLA-DR; however, this appears to be restricted to a subset of B lymphoblasts.


Surface changes on dental materials. II. The influence of two different dentifrices on surface roughness measured by laser reflexion and profilometer techniques. Surface changes on eight different dental materials were investigated, using laser reflexion technique and a profilometer. The tested materials were: Acrylic veneers Isosit, Isosit N, K+B+, Multiplus and K+Bpaste Gold Sjödings C Titanium pure non-rolled titanium Ceram a potential ceramic direct filling material The materials were subjected to brushing in a brushing machine, with a toothpaste-water slurry. Two different toothpastes were used separately; Clinomyn and Colgate. The new ceramic material showed no measurable wear by the brushing. All the other materials were affected to some extent. On titanium increasing values of light intensity were obtained thus indicating a smoother surface. Among the veneer materials K+Bpaste was the most wear resistant.


Effect of heparin and enoxaparin on platelet interaction with fibrin clots. It has been recently shown that platelet-rich thrombi are particularly resistant to thrombolysis. Since unfractionated heparin was reported to enhance fibrinogen binding to platelets responsible for the hyperaggregating effect of this drug, the purpose of this work was to determine whether or not heparin could also modulate platelet interaction to whole blood clot. We have therefore investigated the retention to a standard clot of 111In-labelled platelets suspended in plasma in presence of saline (control), heparin or a low molecular weight heparin (Enoxaparin). We have shown that the platelet interaction to the clot was significantly increased by heparin but not by enoxaparin used at the same anti Xa activity. In conclusion, this difference may favor the use of enoxaparin over heparin in clinical situation associated with platelet retention to fibrin clot, such as thrombolysis.


Changes in thrombomodulin level in plasma of endotoxin-infused rabbits. Changes in the plasma thrombomodulin (TM) level were examined in endotoxin-infused rabbits. The plasma TM level in normal rabbits was 143.8 +/- 8.4 ng/ml (n = 67) and the molecular weight of the major TM was about 55 kd. Endotoxin (lipopolysaccharide, LPS, E. Coli B8:0127) was intravenously infused. LPS infusion increased the plasma TM level dose-dependently between 0.2 mg/kg and 5 mg/kg. When 5 mg/kg LPS was infused, the plasma TM level started to increase immediately and was 2.3 times higher than the control value within 1 hr. The molecular weight of the major TM was about 75 kd. This rapid increase in TM occurred before the decrease in fibrinogen content and the prolongation of prothrombin time. To examine the effect of circulating leukocytes on the TM increase in endotoxin-infused rabbits, 5 mg/kg LPS was infused into rabbits with leukocytopenia induced by X-ray irradiation. The maximum plasma level of TM was significantly lower than in the untreated rabbits given LPS. These data suggest that the increase in plasma TM is caused by LPS-stimulated leukocyte's prior to hemostaseological changes. It is well known that endothelial cells can be injured by stimulated leukocytes, so this increase in plasma TM probably reflects the deterioration of endothelial cells. This deterioration decreases the ability of endothelial cells to inhibit thrombosis, which would, in turn, contribute to the development of disseminated intravascular coagulation in endotoxin-infused rabbits.


Reduced guanine nucleotide-stimulated polyphosphoinositide specific phospholipase C in platelet hyperaggregation in IDDM. Platelet activity is increased in persons with insulin dependent diabetes mellitus (IDDM). Receptor-medicated phospholipase C (PLC) activation and hydrolysis of phosphatidylinositol bisphosphate (PIP2) accompanies platelet activation. Previous work from our laboratory has shown that PIP2 hydrolysis is decreased in platelets of persons with IDDM. PIP2 hydrolysis is mediated via a phosphoinositide(PI)-specific PLC. PI-PLC activity is regulated by guanine nucleotide(GTP)-binding proteins. We therefore examined the hypothesis that platelet aggregations and PI turnover in platelet from subjects with IDDM is linked to alterations in PI-specific PLC activity. We found thrombin induced platelet aggregation was increased in the IDDM group. Basal PI and PIP2-specific PLC activity was not statistically different for the two groups. Guanine-nucleotide stimulated PIP2-specific PLC activity was decreased in the IDDM platelets. The mechanism for the reduced PLC activity and its role in the platelet hyperaggregation requires further study.


Retroelements in higher plants.

Retroelements in higher plants. Representatives of several classes of retroelements have been characterized in a broad range of plant species, where they appear at variable and sometimes very high copy numbers. So far, only a very small number of plant elements have been shown to be active, and this activity seems to be restricted to specific situations of 'genomic shock'. Although it is not yet known whether the presence of retroelements is linked to the high level of variability found in plant genomes, it is now clear that retrotransposons are ancient and ubiquitous components of plant genomes, and could play an important role in plant evolution.


[Preclinical and prenatal diagnosis of familial adenomatous polyposis]. In order to investigate the possibility of preclinical and prenatal genetic diagnosis of familial adenomatous polyposis (FAP) by means of DNA-systems and other markers, blood samples were collected from 246 persons in 29 families, including 90 with the clinical diagnosis FAP and 73 clinically unaffected first degree relatives (persons at risk). The material was studied with up to 4 DNA-marker systems located in the region around the disease gene. Among the first degree relatives eight (11%) had probably inherited the disease gene, while 31 persons (42%) in this risk group had probably not inherited the gene. It was not possible to evaluate the risk in the remaining 34 persons (47%). In 45 (85%) out of 53 persons under 40 years the DNA-systems were informative, so that it would be possible to offer the option of prenatal diagnosis. It is concluded that preclinical and possibly prenatal genetic diagnosis may be offered; but the current practice of prophylactic proctosigmoidoscopic surveillance should be maintained.


Unusual primary prostatic malignancies.

Unusual primary prostatic malignancies. Atypical prostatic cancer is defined as any malignancy of prostatic origin, excluding the common acinar prostatic adenocarcinoma. These atypical prostate cancers account for less than 5 percent of all prostatic malignancies. Because atypical prostatic malignancies occur rarely, little is known about their biologic behavior. To date, no large series of patients have been reported, and specific treatment protocols have yet to be defined. Herein we describe 5 new cases of atypical prostatic cancer. By reporting the occurrence of new cases, we believe a better understanding of the natural history of these lesions will emerge.


Small cell carcinoma of prostate: effectiveness of hormonal versus chemotherapy. Small cell carcinoma of the prostate is rare and associated with a rapidly fatal course. Since 1977, 47 cases have been reported in the world literature with data from 3 additional cases presented herein. The purpose of our review was to determine the effectiveness of hormonal versus chemotherapy. Thirty-four of the 50 cases have known clinical histories. Four patients were not treated, and all were dead of their disease within an average of 2.75 months. Six patients were eliminated from our review because small cell carcinoma was discovered at autopsy. Another 5 cases were omitted because hormonal +/- chemotherapy had already been given for a previous diagnosis of adenocarcinoma, but no specific therapy was given once the small cell carcinoma developed. Of the remaining 19 cases, only 2 have survived. One is still alive forty-three months after hormonal treatment, and another is alive with disease six months after the initiation of hormonal therapy and chemotherapy. Five patients were given hormonal therapy only, and none of them responded. In 4 patients chemotherapy was given after hormonal therapy had failed, and they too died of their disease within a short period of time. However, an additional 8 patients were treated with immediate chemotherapy +/- hormonal therapy and had substantially longer clinical remissions. Therefore, although small cell carcinoma is a uniformly fatal disease, immediate chemotherapy should be considered to promote better clinical remissions.


Testicular ultrasonography after chemotherapy for germ cell neoplasm: significance of highly hyperechoic lesions. The sonographic appearance of the testis after administration of chemotherapy for metastatic germ cell neoplasm is not well known. Fifty-six patients (60 testes) who were previously treated with chemotherapy for metastatic germ cell neoplasm (originally diagnosed by removal of the contralateral testis or by biopsy of metastatic disease) underwent sonography followed by orchiectomy. The sonographic characteristics found to predict viable intratesticular tumor were: lesion size larger than 5 mm, fewer echoes than adjacent parenchyma (hypoechoic), inhomogeneous echo texture, poor margin definition, cystic areas, or highly hyperechoic foci within a hypoechoic lesion. Fibrosis was predicted by finding single or multiple small, highly hyperechoic lesions. These results suggest the potential for predicting the pathologic diagnosis in some patients after receiving chemotherapy for germ cell neoplasm.


Herpetic esophagitis in an immunocompetent boy. An immunocompetent 12-year-old boy had epigastric pain, odynophagia, fever, and hematemesis. Esophagoscopy demonstrated ulceration of his proximal and distal esophagus. Although histopathologic analysis of biopsies obtained from the lesions was nondiagnostic, herpes simplex virus type 1 was isolated from these samples. The patient responded rapidly to acyclovir therapy. Only a handful of cases of herpes esophagitis have been reported previously in immunocompetent children. This infection must be considered in children having odynophagia and fever, even if they are not immune compromised.


[Prolonged posttetanic changes in the sensorimotor cortex of the waking rabbit in response to stimulation of the fibers of the substantia alba in the neocortex and corpus callosum]. Study of posttetanic changes in the sensorimotor cortex of the alert rabbit in response to stimulation of callosal fibres of the white matter have shown that in comparison to the control level either a prolonged increase (potentiation) of the tested evoked potentials to the stimulation of the same structures takes place, or the amplitude of the tested evoked potentials decreases (depression). Both processes can take place simultaneously in different layers of the cortex.


[The thalamic integration of afferent flows in man during image recognition]. The article deals with the mechanisms of interaction of afferent flows at intrathalamic and thalamocortical levels of integration in humans identifying visual, auditory and tactile images. As the methodic model of thalamic block we used patients of surgical clinic with topically diagnosed volume process of the region of the third ventricle. Psychophysiological tests and multichannel EPs recording revealed functional significance of thalamus in the processes of perception and also mechanisms of compensation of these functions in pathological process.


[The characteristics of trace rhythm reproduction by the neurons of the rabbit sensorimotor cortex in the aftereffect of periodic stimulation]. Trace rhythm recruitment (TRR)--CR analogue to time was studied appearing in response to prolonged electrocutaneous stimulation of the forelimb of the alert rabbit with the frequency 0.5-1-2 Hz. The activity was recorded of 180 cells of the sensorimotor cortex before (80) and after (100) periodical stimulation during 10-20 min. The first series of rhythmic stimulation led to a short-term TRR of the stimulation frequency, the following series formed a clear TRR, preserved for several days. The possibility was revealed of "relearning" of neurones at stimulation rhythm change. The ability of TRR phenomenon of extinguishment, prolonged preservation and reproduction of traces, "relearning" brings it nearer to the processes, analogous to the temporal connection. The ability to reveal distinctly and to quantitatively estimate the characteristics of the applied stimulus fixated by the neurones, makes this model perspective for comparable study of the memory traces at the neuronal level in the animals of various ages.


[The interhemispheric brain asymmetry of the rabbit in a state of "animal hypnosis"]. By the method of forced immobilization the rabbits were brought into the state of "animal hypnosis" (immobilization reflex), and their ECoG was recorded, which was further processed on the computer. It was found that during hypnosis a functional interhemispheric brain asymmetry was developed in rabbits with activity predominance in the right hemisphere. The "animal hypnosis" is a phasic process: in the ECoG of the rabbit under hypnosis a regular alternation of delta and theta activity takes place. Electrophysiological reconstructions in the rabbit brain during the change of its functional state correlate with the brain thermal reactions, revealed earlier.


[The differential sensitivity to a neurotigenic exposure of rat strains differing by the threshold of nervous system excitability]. The influence was studied of 15-days stressing on the appearance of stable neurosis-like state of rats lines, selected by the excitability of the nervous system. Unconditioned and conditioned components of behaviour were tested: pain sensitivity, behaviour in the open field, level of "anxiety", passive and active defensive avoidance. Differential reactivity was shown of the rats lines to prolonged stressing, depending on the genetically determined level of the nervous system functional state. Interlinear differences in dynamics of the development of neurosis-like state were established.


[The characteristics of shuttle avoidance learning in spontaneously hypertensive and normotensive rats]. In spontaneously hypertensive (strain SHR) and normotensive (strain WKY) rats was studied the elaboration of conditioned reflex of active avoidance in shuttle box. In case when the shuttle box was divided by a partition the SHR rats learned worse than WKY rats. In shuttle chamber without partition the SHR rats, on the contrary, learned better that WKY ones. Such character of interlinear differences can be connected with properties of formation of the instrumental habit of deliverance from electropainful stimulus, because the presence of partition significantly hampered its fulfillment. The obtained results, compared with literature data, testify to the fact that differences of SHR and WKY rats in elaboration of conditioned reflexes are explained basically by the properties of their unconditioned activity and not of the associative processes.


[A dipole localization method in analyzing the auditory brain-stem evoked potential]. The localization of generators of brainstem auditory evoked response (BAER) has been studied in one patient using the method of dipole localization based on the spatial distribution of BAER over the surface of the head. It has been found that in formation of all BAER waves the activity of several generators overlaps. It is especially marked for the peaks I', II', III and III'--their potential distribution can not be described by single-dipole model, thus preventing the defining of their generation site. Distribution of potential for other peaks corresponds to the single-dipole model. The coordinates of the equivalent sources of these peaks are in the vicinity of the auditory structures: I--near the distal part of the auditory nerve, II--near the auditory nerve in the place of its entering the brainstem, V--near contralateral auditory pontine structures, V--near lateral lemniscus while V', VI and VI'--near mesencephalic auditory structures.


[Changes in the matching of somato-autonomic rhythms in the human operator during learning]. In subjects during the 1st through 15th day of training of the assemble job, before and during the work, the changes of concordance between the heart rate and respiration rhythm were studied in comparison with the dynamics of the activity results and of psychophysiological characteristics. In subjects with bad results of learning higher indices of respiratory-cardiac concordance and tremor were initially observed, what may serve as a criterion for professional selection for this kind of activity. In well learning subjects during transition from rest to work the initial desynchronization of somato-vegetative rhythms ("stress" type of reaction) was changed by an increase of their concordance. The relation of the training success to the type of working capacity of the subject ("morning", "evening") was also revealed.


[The age-related characteristics of the correlation of involuntary and voluntary analysis during image recognition]. An experimental paradigm was used, based on the stimuli presentation in the absence and presence of the attention, directed to them; it allowed to divide involuntary (automatic) and voluntary (controlled) processing levels of the analysis, functioning parallelly in the process of real image identification. In conditions of this paradigm the regional and hemispheric organization of the evoked electrical activity (ERP and differential curves) was studied in youths aged 16-17 (11 persons) and children aged 7 (15 persons). The analysis of the differential curves revealed periods of negativation and positivation, connected with the automatic (unattended stimuli) and controlled (attended stimuli) analysis of images. Correlation of the identification success and singled out periods of differential curves has shown that in children aged 7 sensory decisions achieved on the automatic level are a significant contribution to the identification of images. Attraction of the attention to the stimulus in children aged 7 intensifies its processing, observed already after making the main decision on the character of the stimulus. In youths the success of identification correlated mainly with the processes at the controlled level; in this case the integrating role of the left frontal area at the final stage of the analysis is singled out.


[Narrow-band changes in the the EEG spectral composition under the influence of an agonist and an antagonist of noradrenergic neuromediation]. At pharmaco-electroencephalographic examination of patients with neuroses narrow-band components was carried out of the EEG spectrum, which oppositively changed under the influence of sidnocarb and obsidan--agonist and antagonist of noradrenergic system, the initial hypothesis on the perspectivity of search of the EEG markers of the level of activation of the brain neurotransmitter systems thus being confirmed. Comparison of the obtained EEG-data with pharmacological properties of the applied drugs allows to project the aspects of further EEG studies of neurotransmission.


Pathomorphological and immunohistological findings in progeny of goats experimentally infected with pestiviruses. A total of 25 pregnant goats without neutralizing antibodies against BVD virus were inoculated with two different pestivirus isolates at eight different stages of gestation. In both infection groups, various malformations were observed in fetuses and neonates. In three twins with neutralizing antibodies against BVD virus leukoencephalomalacia occurred, characterized by gelatinous transformation in the cerebral hemispheres. These lesions were comparable to alterations described in alternative pathology of Border disease in sheep. Although the immunohistological findings are characteristic for immunological tolerance and viral persistence, viable offspring persistently infected with pestivirus was not observed.


[Changes in the late components of evoked cortical potentials during the human solving of a visual-spatial task]. The ability of mental construction of easily verbalized geometric figures was studied in 20 healthy subjects by means of recording of the evoked potentials of the cerebral cortex. Fragments of geometric figures were presented with the intervals of 80 and 500 ms. To one half of the subjects the stimuli were presented unilaterally (both fragments to one and the same visual field, to the left or to the right one) and to the second half of the subjects--bilaterally (the first fragment--to one visual field and the second one--to the other). It was established that the latency of the wave P300 was shorter and the amplitude--greater at correct decisions of the visual-spatial task than at false ones, both at unilateral and bilateral fragments presentation. At a great interval (500 ms), when the effects of masking became significantly weaker, a predominant activation of the left hemisphere was observed: the latency of the late components was shorter and the amplitude--greater, when the information came "directly" to the left hemisphere. The obtained results testify once more to a great variability, dynamic interhemispheric relations, impossibility of connecting the realization of some functions with one of the hemispheres.


[The acquisition in dogs of temporal regulation by differentiated reinforcement of the response duration using an exteroceptive component and the pharmacological modulation of this reflex (diazepam, clozapine)]. Transformation of expectation phenomenon into the phenomenon of temporal regulation is usually achieved by the suppression of preliminary factors or by means of their physical modification. Our studies show that such transformation can be obtained in dogs using Kupalov paradigm with the presentation of additional stimuli. These stimuli strictly identical, from the physical point of view, to the signals which interrupt expectation are randomly introduced into the temporal limit. Absence of the reinforcement in response to the additional stimulus impels the animal to include temporal regulation in its behaviour, and an additional negative discriminative stimulus promotes an expression of active character of inhibition. These circumstances make our pattern closer to DRRD (differential reinforcement of response duration). In order to evaluate the merits of this procedure the influence was studied of anxiolytic (diazepam) and neuroleptic (closepine) on the stabilized reaction of experimental animals. The increase of responses duration by closepine and their shortening by diazepam as well as the influence of these pharmacological substances on the frequency of responses in dependence of dose, confirm the results of the previous studies of DRRD and DRL (differential reinforcement of low rate of responses) and prove differential sensitivity of our procedure to pharmacological substances.


An immunoblotting procedure for detection of antibodies against bovine leukemia virus in cattle. A sensitive protein immunoblotting (Western blot) procedure has been developed for detecting anti-BLV antibodies in cattle sera. The antibodies against most of the major viral proteins could be detected. This procedure does not give any non-specific background staining and there is absence of any erroneous results due to utilisation of purified viral preparations. The procedure has been applied for detection of antibodies to BLV in a set of 74 sera samples and it has been compared with other commonly used serological tests like ELISA and agar gel immunodiffusion test.


Prevalence of feline leukemia virus and antibodies to feline immunodeficiency virus in cats in Norway. Serum samples from 224 Norwegian cats were analyzed for the presence of feline leukemia virus (FeLV) p27 common core antigen, and for antibodies to feline immunodeficiency virus (FIV). Ninety specimens originated from the serum bank at the central referral clinic at the Norwegian College of Veterinary Medicine, which had been collected during the years 1983-1989; 67 sera were submitted from veterinarian practitioners; while 67 sera originated from cats presented for euthanasia. The cats were classified into one "healthy" and one "sick" group. Only 2.2% of sick cats and 1.2% of healthy cats showed FeLV antigenemia, a finding which is lower than which has been reported from many other countries. The prevalence of FIV antibodies was 10.1% in sick cats and 5.9% in healthy cats. Antibodies to FIV was most prevalent in male cats (14.7%) than in female cats (2.1%), and more prevalent among domestic cats (12.0%) compared to pedigree cats (2.4%). Antibodies to FIV in the cats demonstrated increasing prevalence with increasing age. It may be concluded that FeLV causes minor problems in Norwegian cats, while FIV is present in a similar prevalence to what is reported from other countries.


[The psychophysiological testing of temperament]. A comparison was carried out of the dynamics of the activation focus of the human cerebral cortex with the data of psychological testing by the ergic parameter of the "Questionnaire for estimation of temperament structure". It is shown that the right hemisphere in the subjects conditionally attributed to the sting type ("highergice") is much more involved involving verbal tasks than in "week" types. In subjects conditionally attributed to the "week" type, at fulfillment of verbal tasks inert predominance of the left hemisphere activation is noted.


[The assessment of the strength of the interaction between neurons of the cat motor cortex in a conditioned reflex to time]. The work was conducted on cats with recording multineuronal activity of the motor cortex at the elaboration of conditioned reflex to time. Strength of interaction was estimated between the adjacent and remote neurones in the limits of 0.5 m. The dynamics of strengths of interneuronal interaction in most cases did not correlate with the dynamics of impulses frequency of the studied neurones. Changes of strengths of interaction between mutually remote neurones at CRT were met more frequently than between the adjacent ones, what may serve as one more evidence of the hypothesis on more strict structure of connections within microsystems and greater plasticity of connections between microsystems.


Direct detection of bovine leukemia virus infection: practical applicability of a double polymerase chain reaction. A double polymerase chain reaction (PCR) assay has been devised for the direct detection of bovine leukemia virus (BLV). The assay was directly performed on blood leukocytes, avoiding the DNA-purification procedures. The PCR products were identified by gel-electrophoresis and the specificity of the test was confirmed by hybridization with a biotinylated oligonucleotide probe. When testing the sensitivity of PCR, less than eight genome copies of the provirus were detected in the background of two million negative lymphocytes. In a BLV infected herd 22 animals of various age groups were examined by the indirect (serological) diagnostic tests of agar-gel immunodiffusion and indirect ELISA as well as by the direct detection method of PCR. The tests were repeated at monthly intervals on five occasions. When examining the specimens from cows and heifers, a close agreement was found between the results of the various methods. The newborn calves, which were the offspring of BLV infected mothers, were consequently negative in PCR throughout the experimental period. However, in the indirect tests the calves were positive during the first samplings and became negative only around four months of age. Since the indirect tests can not discriminate infection from colostral immunity, PCR proved to be a useful complementary assay for the safe diagnosis of BLV infection in young calves.


[The formation of a complex habit in Black Sea bottle-nosed dolphins under free-choice conditions]. In experiment situation was created of "free choice", which was characterized by the reinforcement of any succession of bottle-nosed dolphin action on the manipulators, placed in the open-air cage under the condition of pressing three definite levers in the required order independently of the number of "superfluous" reactions before and between necessary actions. In the first series of the experiments with eight above-water levers no succession of actions leading to the reinforcement was formed in the bottle-nosed dolphin [correction of aphaline]. In the second series with eight under-water manipulators--both experimental dolphins regularly reached the reinforcement. Stereotype trajectories of movements in the cage were formed, which included swimming past the levers and successive actions on them. Minimum chain of motor reactions was not formed, though some of "superfluous" levers were not pressed. At solving the task efferent generalization in dolphins in both series of experiments was manifested in two ways: the animals reacted in different ways on one and the same manipulator and pressed different levers.


[The theta modulation of rabbit hippocampal neurons and its correlation with other indices of spontaneous and evoked activity]. Reliability of the existing functional criteria for differentiation of pyramidal ("complex spike neurones") and inhibitory ("theta neurones") cells in the hippocampus of waking rabbit is evaluated on the basis of statistical analysis of neuronal spontaneous and evoked activity. The analysis shows, that the criteria of mean frequency, presence of theta modulation, neuronal behaviour in situations provoking EEG theta rhythm (e.g., excitation or inhibition during presentation of sensory stimuli), effects of medial septum and intrahippocampal stimulation do not permit reliable identification of the hippocampal neuronal types in the waking rabbit. The data on functional classification of the hippocampal neurones are discussed in connection with existing suggestions about their state in situations inducing theta rhythm generation.


[Evoked potentials of the brain limbic structures during a change in the motivational state of dogs and during the realization of unconditioned reflex reactions]. In experiments on 6 dogs it has been shown that in the EPs led from the hippocampus in response to electrostimulation of limbic structures and from the latters at the hippocampus stimulation, initial negativity, after positivity and late negative wave are the most stable components. At transition from one functional state to another two latter waves are subjected to the greatest changes. Amplitude-temporal characteristics of intralimbic EPs depend on the level and type of motivation, degree of emotional stress and properties of conditioned reaction.


Pulmonary sclerosing hemangioma. Report of a case with diagnosis by fine needle aspiration. Sclerosing hemangioma is a rare but well-recognized benign lesion of the lung. We report a case of pulmonary sclerosing hemangioma correctly diagnosed by fine needle aspiration (FNA) cytology. The sharp and smooth contour of the discrete mass in the left lower zone of the chest roentgenogram raised the possibility of a benign lesion, including pulmonary sclerosing hemangioma. The characteristic "blood spaces" with surrounding regular, bland polygonal tumor cells in the FNA smears provided an essential clue to the diagnosis of sclerosing hemangioma. It was confirmed by Surecut biopsy of the lesion. The patient remained well one year after the investigation and was spared an unnecessary diagnostic thoracotomy. The cytologic features and differential diagnoses of pulmonary sclerosing hemangioma are discussed. Besides delineating the cytologic characteristics of pulmonary sclerosing hemangioma, this case illustrates the importance of a careful clinicopathologic correlation, which should be exercised by the cytopathologist in all instances.


Pseudomyxoma peritonei associated with ovarian mucinous tumors. Cytologic appearance in five cases. The cytologic findings in five cases of pseudomyxoma peritonei associated with ovarian mucinous tumors are reported. The evidence suggests that if mucinous or gelatinous ascitic fluid is received in the laboratory and is shown to contain a dual cell population of round cells of mesothelial origin and spindle-shaped cells of fibroblastic origin together with lakes of fibrillar mucin, a cytologic diagnosis of pseudomyxoma peritonei can be made with confidence. It will usually indicate mucinous neoplasia of the appendix or ovary. In the case of ovarian neoplasia, the tumor is most likely to be a low-malignant-potential mucinous tumor. The patient's prognosis, however, will be that of mucinous carcinoma rather than of usual borderline tumors of the ovary.


Are oral contraceptive-associated liver cell adenomas premalignant? From January 1977 to June 1990, 1,670 patients with a liver mass or masses underwent transabdominal fine needle aspiration biopsy of the liver. Of those cases, 99 were diagnosed cytologically as "hepatocellular carcinoma" and 9 as "consistent with liver cell adenoma." Among the 99 patients with hepatocellular carcinoma, 3 were users of oral contraceptives. The nine patients with liver cell adenoma were all users of oral contraceptives. Of them, two developed foci or areas of liver cell dysplasia within the adenomas. The mean periods of oral contraceptive use among these three groups of patients were different. It appeared that patients started to develop liver cell adenoma after five years of oral contraceptive use (mean, 6.3 years). Foci or areas of liver cell dysplasia began to arise within liver cell adenomas after 8 years of oral contraceptive use (mean, 9 years), and the patients started to develop hepatocellular carcinoma after 10 years of contraceptive use (mean, 11 years). The cytologic features of liver cell dysplasia strikingly mimicked those of hepatocellular carcinoma. From this study, the foci or areas of liver cell dysplasia appear to be the missing link responsible for the transformation of liver cell adenoma to carcinoma. It is believed that liver cell adenomas are not premalignant and may undergo reversible change after withdrawal of causative agents, whereas foci or areas of liver cell dysplasia within the adenomas are irreversible, premalignant changes and may transform into hepatocellular carcinoma.


The comedo subtype of intraductal carcinoma. Cytologic characteristics. Smears from 10 intraductal carcinomas of the comedo type without microinfiltration were compared with smears from 10 similar tumors with suspected or proven microinfiltration and smears from 10 invasive comedo carcinomas. Microinvasive tumors could not be separated from purely intraductal cases unless tumor cell infiltration in mammary fat was seen in the smear. The tumor cells in most of the intraductal cases were shed in cohesive groups and clusters, lying in necrotic cellular debris and with few or no scattered, single, dissociated tumor cells. Smears from invasive comedo carcinomas invariably showed tumor cell clusters and scattered, single, dissociated tumor cells, often with atypia in excess of what was seen in the intraductal cases. Also, in most cases the invasive tumors did not show a background of necrotic cellular debris.


Fine needle aspiration biopsy diagnosis of adenoid cystic carcinoma of the breast. A case report. A case of adenoid cystic carcinoma arising in the breast is presented. The diagnosis was established on a fine needle aspiration (FNA) biopsy sample with histopathologic corroboration. The aspirate yielded a typical smear composed of round, multilayered clusters of uniform epithelial cells arranged around cores of homogeneous, acellular material. Both the conclusiveness of FNA biopsy in diagnosing this rare mammary neoplasm and its resulting in immediate treatment are stressed.


Modified cerium-based and Gomori-based cerium methods for light microscopic phosphatase histochemistry: the cerium-perhydroxide-diaminobenzidine-nickel (Ce-H2O2-DAB-Ni and Ce/Ce-H2O2-DAB-Ni) two-step procedures. Some modified cerium-based and Gomori-based cerium methods for the demonstration of phosphatase activity in cryostat sections were described. Dextrane as stabilizing agent was added to the incubation media for ATPase, 5'-Nase, and TPPase. The oxidation of the CeIII-phosphate primary reaction product in a separate step by H2O2 before the DAB incubation yielded an increase of the intensity of the DAB-based visualization reaction (Ce-H2O2-DAB-Ni two step method). The sensitivity of the histochemical enzyme reaction was remarkably increased if CeIII-ions were employed as amplifying agent (Ce/Ce-H2O2-DAB-Ni two-step method). A new suitable DAB medium consisting of 0.015% DAB, 2.0% Ni-sulphate, 15% methanol, and 0.005% H2O2 in 0.1 mol/l acetate buffer, pH = 5.2, was used. The disadvantage of diffuse background staining has been overcome by addition of 15% methanol to the DAB solution. Electrovalently bound CeIII (cerophilia) was removed by treatment of the incubated sections with CeIII-citrate (CeIII-complexation). In addition, a novel membrane floating incubation for sections is proposed. At present, the modified procedures are some of the most sensitive modes for the demonstration of phosphatases and improve the earlier described cerium-DAB one-step technique (Halbhuber et al. 1988b).


Spontaneous infarction of a parotid gland pleomorphic adenoma. Report of a case with cytologic and radiographic overlap with a primary salivary gland malignancy. Pleomorphic adenoma is the most common neoplasm of the parotid gland, generally presenting as a slowly enlarging, firm, well-circumscribed, painless nodule. Occasional cases have presented after a short period of rapid growth or have been associated with pain. The vast majority of these tumors are solid, but rare examples have been associated with cystic degeneration or hemorrhage. Spontaneous and tumor-associated infarction of the parotid has been reported, but these examples have been limited to infarctions of Warthin's tumors and postoperative infarctions of salivary glands. We present the case of a 48-year-old male with a one-year history of a painful, enlarging, left parotid mass associated with paresthesia of the tongue. Computed tomographic examination of the parotid demonstrated a left superficial lobe mass with a rim of enhancement and low attenuation center. Fine needle aspiration yielded necrotic debris and atypical squamous elements that were thought to be compatible with carcinoma. A superficial parotidectomy with intraoperative frozen section revealed a pleomorphic adenoma with extensive central necrosis. To our knowledge, this represents the first reported case of an infarcted pleomorphic adenoma and illustrates the potential for misinterpretation of these cytologic and radiologic findings as indicative of malignancy.


A registry study of very low birthweight liveborn infants in Sweden, 1973-1988. Very low birthweight liveborn infants (less than 1,500 g, VLBW) born in Sweden 1973-88 were identified from the Medical Birth Registry and efforts were made to remove wrongly recorded birthweights--9% of infants with a registered birth weight below 1,500 g were removed. Some VLBW infants were not recorded in the register and the estimate of the prevalence at birth of VLBW infants is therefore slightly underestimated. It increased from about 5.5 per 1,000 during the period 1973-84 to 6.7 per 1,000 during 1987-88. 18% of VLBW infants were involved in multiple births. Median Apgar score at 5 min increased for each 100 g birthweight class. Even at a birthweight between 1,400 and 1,499 g, a low Apgar score at 5 min was seen in 20%. The rate of cesarean section increased between 1973 and 1983 from about 10% to 60%. One-year survival for infants with a birthweight less than 1,000 g increased from less than 20% in 1973-75 to 50% in the 1986-88 cohorts. Corresponding figures for infants with a birthweight between 1,000 and 1,499 g were 60% and 90%. A markedly better one-year survival is already evident in the 600-699 g class. On stratifying for 100 g birthweight class, perinatal death risk was higher in boys than in girls and higher in multiple births than in singletons. An increased rate of congenital malformations was seen in the 1,000-1,499 g class but not in the less than 1,000 g class.


Colloid carcinoma of the breast with concomitant metastasis and a tuberculous lesion in the axillary lymph nodes. A case report. A 30-year-old woman presented with a lump in the left breast and left axillary lymphadenopathy that, on fine needle aspiration cytology (FNAC), proved to be duct cell carcinoma with metastasis. Histology of the radical mastectomy specimen showed a mixed colloid carcinoma. Axillary lymph nodes revealed a variety of pathologic changes consisting of reactive hyperplasia, tuberculosis and metastasis. A combination of a tuberculous lesion and metastasis in the same lymph nodes was also found. During follow-up, after radiotherapy, the patient developed left supraclavicular and right cervical lymphadenopathy that, on FNAC, revealed a tuberculous lesion and metastasis, respectively. The rarity of this condition with double pathology is highlighted, and the reason behind the limitations of FNA in subtyping the primary malignancy and its failure to detect the tuberculous lesion in the axillary lymph node are discussed.


Antiphospholipid antibodies in pre-eclamptic women: relation to growth retardation and neonatal outcome. The significance of antiphospholipid antibodies in pre-eclamptic women has not been thoroughly elucidated. The purpose of this study was to determine the proportion of pre-eclamptic women who were antiphospholipid antibody positive, and to elucidate the significance of these antibodies regarding growth retardation and neonatal outcome. Positive levels of anticephalin antibodies, which are antiphospholipid antibodies, were detected in 7 (19%) out of 37 pre-eclamptic women, as compared with none of 40 in a control group of normotensive women at similar stage of pregnancy (p = 0.004). The birthweight percentiles of the neonates of anticephalin antibody positive women were significantly lower than those of the neonates of anticephalin antibody negative women (p = 0.018). Four of 7 infants of anticephalin antibody positive women were growth retarded (less than 2.5th percentile). This was a significantly larger proportion than that for anticephalin antibody negative women (3/30) (p = 0.004). The 95% confidence interval for the difference between the two proportions was 0.10 to 0.85. Two of the 7 neonates of anticephalin antibody positive women died during the neonatal period, compared with none of the 30 neonates of anticephalin antibody negative women (p = 0.003). Thus, our study suggests that positive levels of anticephalin antibodies in pre-eclamptic women increase the risk for growth retardation and neonatal death.


Electrical injury from Tasering and miscarriage. A case report is presented of a woman who was "Tasered" by law enforcement personnel while 12 weeks pregnant. The Taser (Thomas A. Swift's Electric Rifle) is an electronic immobilization and defense weapon that has been commercially available since 1974. The Taser was developed as an alternative to the .38 special handgun. The patient was hit with Taser probes in the abdomen and the leg. She began to spontaneously miscarry 7 days later and received a dilatation and currettage procedure 14 days later for incomplete abortion. The world's literature on electrical and lightning injury to pregnant women is reviewed, and the mechanism of action of Taser injury is discussed. As use of the Taser becomes more common, obstetrical clinicians may encounter complications from the Taser more often.


Fine needle aspiration cytology of mesoblastic nephroma. A case report. A case of congenital mesoblastic nephroma of the left kidney was diagnosed in a 4-month-old child by fine needle aspiration cytology. The smears consisted of clustered and dyshesive spindle cells with minimal nuclear atypia and mitosis. No epithelial, tubular or glomeruloid differentiation was noted. Considering the age and cytomorphology, a diagnosis of mesoblastic nephroma was made. Histopathology of the nephrectomy specimen showed a tumor with features of atypical mesoblastic nephroma. Cytologic diagnosis of mesoblastic nephroma is important because the tumor has an excellent prognosis, and unlike Wilms' tumor, requires only surgery.


Prepartum transabdominal amnio-infusion for severe oligohydramnios. Transabdominal amnio-infusion preceding labor induction was evaluated as a means of avoiding fetal distress and cesarean delivery in patients with oligohydramnios. A preliminary study was performed in 8 consecutive term or post-term pregnancies complicated by severe oligohydramnios (amniotic fluid index less than or equal to 1 cm) with unripe cervices (Bishop's score less than or equal to 3). Warm saline was injected through a spinal needle under ultrasound control. Vaginal delivery occurred in 7 cases; cesarean section was performed in one patient for failure to progress. There was no meconium aspiration and no sign of fetal distress.


Sexual function and somatopsychic reactions after local excision of vulvar intra-epithelial neoplasia. Eighteen patients, under the age of 60 years, who have been treated with local excision of the vulva, participated in the study in which sexual function and somatopsychic reactions were evaluated by personal interviews. Furthermore both objective and subjective cosmetic results were registered. After this type of operation, fewer than one-third of the patients had postoperative sexual and somatopsychic problems, whereas more than half of the patients undergoing vulvectomy did report such problems. Fourteen out of 18 patients were satisfied with the cosmetic result and in 12 of the patients no disfiguration was found. This study shows that local excision of intra-epithelial neoplasia of the vulva is far less sexually traumatic than is vulvectomy.