Phosphorus nuclear magnetic resonance studies on the calcium-dependent energy metabolism of rat cerebrum under conditions of increased potassium in vitro. Using phosphorus nuclear magnetic resonance (31P NMR) we studied the biochemical changes that accompany high-K+ depolarization in cerebral tissue in vitro. Rat brain slices (wet weight 4-6 g) were placed in a glass NMR sample tube which was superfused with well-oxygenated Krebs solution at 25 degrees C. The broad signal seen in the spectrum of brain slices almost disappeared after phospholipids were extracted by using chloroform and methanol. Phospholipids in the extracts were identified as phosphatidylcholine and phosphatidylethanolamine. In high-K+ Krebs solution tissue phosphocreatine (PCr) was decreased while inorganic phosphate (Pi) was increased, and in normal solution PCr and Pi recovered to control levels within about 20 min. The extent of changes in PCr and Pi depended on the concentrations of K+. Ouabain or tetrodotoxin did not influence these changes significantly. In calcium-free solution, where Ca2+ was replaced by Mg2+, the changes caused by high K+ were decreased to 40% of that in normal high-K+ solution. When ouabain was further added to the calcium-free solution, the changes were decreased to 10%. This suggested that the increased metabolism in high K+ was caused mainly by calcium-dependent mechanisms and partly by the Na(+)-K+ pump.


Further evidence for calcium permeability of non-NMDA receptor channels in hippocampal neurons. We have previously suggested that kainic acid (KA) increases the fluorescence of a fluoroprobe for Ca2+ via activation of the non-NMDA (N-methyl-D-aspartic acid) receptor in cultured hippocampal cells. The present study provides further evidence to support this suggestion. KA and NMDA increased the fluorescence occasionally in different subcellular loci of a single cell for each agonist, excluding the possibility that KA activated the NMDA receptor. KA elevated fluorescence in the cell whose membrane was voltage-clamped, thus eliminating possible Ca2+ entry through voltage-dependent channels. A monophasic response to KA was found to occur in the cells identified immunochemically as neurons. Even when the cells were cultured at different embryonic stages, KA responsiveness reached a peak at a rather fixed timepoint through the total age of neurons, while NMDA responsiveness continued to increase. This suggests that the Ca(2+)-permeable non-NMDA receptor is expressed according to a genetic schedule, but not to an artifactual effect of cell culture.


Incidence and clinical features of breast cancer in the Auckland region. Data on all new breast cancer cases in the Auckland area during the nine years September 1976 to September 1985 were used to obtain epidemiological information on breast cancer in the Auckland region. Breast tumours were found in 2706 women (300 per year), yielding a lifetime risk of breast cancer of one in 15. No significant difference in breast cancer incidence was detected between European, Maori and Pacific Island Polynesian women. Confidence limits for incidence were wide in the later groups. Fifty-one percent of women presented with intermediate sized (2-5 cm) tumours, and most (66%) were node negative. Eleven percent had evidence of metastatic disease at presentation. When the relationships between race, tumour size, nodal status and metastases were examined, Pacific Island women more frequently presented with large tumours and metastases, whereas Maori women were more frequently node positive. Eighty-five percent of tumours were invasive ductal carcinomas, 55% grade II, 35% grade III, and 10% grade I. Sixty-seven percent of tumours were oestrogen receptor positive (ER+ve) and ER status was significantly related to age; the proportion of ER+ve tumours was greater in older women. Fifty-seven percent of tumours were progesterone receptor positive (PR+ve), and PR distribution was bimodal with age. These data from the Auckland region are similar to breast cancer figures from other western countries, with some ethnic differences in tumour size and frequency of metastatic disease at presentation.


The dawn phenomenon: comparison between normal and insulin-dependent diabetic adolescents. To determine the role of insulin clearance in the dawn phenomenon, we studied 10 adolescents with IDDM in comparison to 10 healthy, matched control subjects reported previously. In diabetics, metabolic clearance rate of insulin was calculated during i.v. infusion of insulin from 0100 to 0430 h and from 0430 to 0800 h (0.17 and 0.33 mU/kg/min, respectively), with a Harvard pump, while maintaining nocturnal euglycemia. In controls, metabolic clearance rate of insulin was calculated from the prehepatic insulin secretion rate based on C-peptide levels. In diabetic and control subjects, plasma glucose, free insulin, and glucagon concentrations were similar and did not change during the dawn period. However, metabolic clearance rate of insulin increased during the dawn period in diabetic (9.42 +/- 0.91 to 19.89 +/- 1.52 mL/kg/min, p less than 0.0001) and control subjects (4.87 +/- 1.11 to 9.30 +/- 1.50 mL/kg/min, p = 0.008). Plasma cortisol and adrenocorticotropic hormone levels increased and growth hormone (GH) decreased significantly during the dawn period. Diabetic adolescents had significantly higher plasma GH levels than control subjects throughout the night. We conclude the 1) increased insulin clearance is responsible for the dawn phenomenon in healthy and diabetic adolescents and 2) insulin resistance due to GH is an unlikely cause for the dawn phenomenon because diabetic subjects, despite higher GH levels, maintain plasma glucose levels similar to control subjects without requiring higher plasma free insulin concentrations.


Effects of prolactin on Na+K(+)-ATPase activity in the nephron during maturation in the rat. The effect of prolactin (PRL) on renal Na+K(+)-ATPase was investigated in 7-d-old neonatal rats. Animals were treated by bromocriptine (Br; a blocker of endogenous PRL secretion), and the enzyme activity was compared with that of untreated controls. Na+K(+)-ATPase was determined in renal sections in the medullary thick ascending limb of Henle's loop and in the distal tubule by cytochemistry. In the distal tubule, Na+K(+)-ATPase activity was significantly lower in Br-treated animals than in controls (330 +/- 169 versus 558 +/- 146 pmol inorganic phosphate/mm/h, respectively); values did not differ in the medullary thick ascending limb of Henle's loop between Br-treated and control animals (132 +/- 74 versus 165 +/- 113 pmol inorganic phosphate/mm/h, respectively). In vitro effects of PRL were investigated by determining the enzyme activity after incubation of renal sections from Br-treated and untreated animals with different concentrations of PRL. Results suggest that PRL may affect renal Na+K(+)-ATPase activity in the distal tubule in the neonatal period but do not support a major role of PRL in the enzyme maturation.


Inotropic responses to selective (RO 20-1724 and SQ 65,442) and nonselective (trequinsin) inhibitors of cyclic AMP-specific class IV phosphodiesterase in newborn, immature, and adult rabbit myocardium. In contrast to myocardium from adult rabbits, myocardium from newborns is insensitive to the inotropic effects of selective inhibitors (e.g. amrinone, milrinone, and indolidan) of the cGMP-inhibited high-affinity cAMP phosphodiesterase (PDE) localized in the sarcoplasmic reticulum. This difference may be explained at least partially by our recent observation that this cAMP PDE activity is low in sarcoplasmic reticulum from newborns. Furthermore, because the predominant cytosolic high-affinity cAMP PDE activity in newborns is a cGMP-insensitive form, we postulated that selective inhibitors of this form of cAMP-specific PDE may increase cardiac contractility in newborns. Therefore, the inotropic effects of RO 20-1724 and SQ 65,442 (selective inhibitors of cGMP-insensitive, high-affinity cAMP PDE) were compared with trequinsin (a potent, less selective PDE inhibitor) in right ventricular papillary muscles isolated from newborn (NB; 24-48 h), immature (14-16 d), and adult New Zealand White rabbits. At a drug concentration of 100 microns, RO 20-1724 and SQ 65,442 depressed maximal rate of tension development to 67 +/- 4 and 70 +/- 2% of control, respectively, in NB papillary muscles. The NB response to RO 20-1724 differed significantly from the immature (127 +/- 2%) and adult (115 +/- 3%) groups (p less than 0.05), but the effects of SQ 65,442 were comparable among the three age groups. In contrast, trequinsin exerted a positive inotropic effect in the NB group (355 +/- 22% of control) that was substantially greater than the maximal response obtained in the immature (139 +/- 6% of control) or adult (131 +/- 5% of control) groups (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


Perception of different frequencies of cranial transcutaneous electrical nerve stimulation in normal and HIV-positive individuals. Sine-wave transcutaneous electrical nerve stimulation (TENS) of varying frequencies applied across the cranium (ear to ear) has been demonstrated to evoke three different noncutaneous sensations in three discrete, nonoverlapping frequency bands in normal, healthy subjects. This report describes two studies which evaluate perception of these cranial TENS-evoked, frequency-dependent sensations in normal and HIV-positive individuals. In Exp. I, all of 50 normal, healthy subjects reported perceiving the same three noncutaneous sensations in the same three nonoverlapping frequency bands as long as stimulated and over repeated trials. In Exp. II, 34 HIV-positive individuals (14 asymptomatic, 9 ARC, 11 AIDS) who were free of neurological symptoms differed significantly from 10 normal, healthy controls, and from the norms observed in Exp. I, on perception of the three different TENS-evoked sensations. Also, inability to maintain perception of the stimulus over repeated trials, observed only in the HIV-positive individuals, increased significantly with severity of HIV infection.


[Liver abscess formation after treatment of liver cancer by arterial injection using adriamycin/mitomycin C oil suspension (ADMOS)]. Of 210 patients with hepatocellular carcinoma (n = 135), metastatic liver cancer (n = 71) and cholangiocarcinoma (n = 4) who underwent intra-arterial infusion of adriamycin and/or mitomycin C oil suspension (ADMOS) and cisplatin, and both regimens, pyogenic liver abscess occurred in seven (3.3%). The percentages of abscess formation in the respective types of liver cancer were 0.8, 7.0 and 25%. These differences among the three types of liver cancer were attributed to the volume of the tumor vascular beds to be embolized, which might determine the relative amount or regional Lipiodol retention in the tumor and normal liver tissue. Four of seven patients with hepatic abscess had received the intra-arterial infusion of ADMOS, and their angiographic findings showed sequential decreases in the vascular beds of the tumor in comparison with those of previous infusion procedures; all had hypovascular liver tumors angiographically. We have never experienced this complication in other treatments such as embolization of the hepatic arteries and intra-arterial infusion of water-soluble anticancer drugs alone. These results suggest that the most important factor leading to abscess formation is the ischemic destruction of the intrahepatic ducts secondary to occlusion of the peribiliary arterial plexus by Lipiodol and/or the direct effects of anticancer drugs on these vessels. To avoid this complication, the volume of Lipiodol used for intraarterial infusion therapy should be carefully determined, especially when the patient has hypovascular tumors of the liver and a history of multiple previous intraarterial infusion procedures of anticancer drug. The use of ADMOS should be avoided in patients with hypovascular tumors of the liver such as secondary deposits and cholangiocarcinoma.


[Clinical usefulness of 99mTc-PMT whole body scans in the diagnosis of bone metastases from hepatocellular carcinoma]. The diagnostic value of whole body scanning using 99mTc-N-pyridoxylmethyltryptophan (PMT) was evaluated in 16 patients with bone metastases from hepatocellular carcinoma, in comparison with 99mTc-MDP. Of the 72 known lesions of bone metastases, 63 (87.5%) were detected by 99mTc-PMT scintigraphy, which demonstrated increased uptake of radionuclide. However, 99mTc-MDP bone scintigraphy detected only 45 lesions (62.5%), which were shown as increased, decreased, or mixed patterns of uptake. Thus 99mTc-PMT scintigraphy was more sensitive than 99mTc-MDP bone scintigraphy. In addition, the latter showed poor specificity because of its high false positive rate due to degenerative change. All lesions undetected by 99mTc-PMT scintigraphy were located in areas that overlapped the liver or bowel activity. In conclusion, it is recommended that whole body 99mTc-PMT scintigraphy be combined with 99mTc-MDP bone scintigraphy for the detection of bone metastases from hepatocellular carcinoma.


The formation of A-DNA in NaDNA films is suppressed by netropsin. Oriented films of NaDNA complexed with netropsin were studied with deuterium nuclear magnetic resonance (2H NMR), X-ray diffraction and ultraviolet (UV) linear dichroism to obtain information about the influence of netropsin on the structural arrangement of the DNA bases and on the B-A transition. The results of these studies clearly demonstrate a strong suppression of the formation of A-DNA at relative humidities (RHs) down to about 50%. The suppression was complete in the NaDNA-netropsin complex studied with 2H NMR which had a netropsin input ratio, r, of 0.22 drug/base pair. The sample used for UV linear dichroism had a similar input ratio while the X-ray diffraction samples had input ratios between 0.033 and 0.39 drug/base pair. Together, the results of these studies are in agreement with previous infrared (IR) linear dichroism studies of the conformation of the sugar-phosphate backbone in NaDNA-netropsin complexes, which showed that the B-A transition is suppressed for r-values down to approximately 0.1 drug/base pair (Fritzsche, H., Rupprecht, A. and Richter, M., Nucleic Acids Res. 12 (1984) 9165-9177).


Herpes simplex virus type 1 polypeptide ICP4 bends DNA. ICP4, the major regulatory polypeptide of herpes simplex virus type 1, is expressed at the earliest stages of virus infection and is required for the activation of transcription from the majority of viral promoters. It is a DNA binding protein which specifically recognises bipartite sites related to the sequence ATCGTnnnnnCGG. In this report we show that both partially purified ICP4, and its isolated DNA binding domain, bend DNA at occupied binding sites. The apparent angles of bend at two different binding sites were very similar and in both cases the centre of the bend was very close to the binding site sequence.


Targeting deletion (homoeologous chromosome pairing locus) or addition line single copy sequences from cereal genomes. We describe here a protocol for obtaining clones containing sequences present in low copy-number from genomic DNA where moderately and highly repeated sequences predominate. Specific chromosomal regions can be targeted by using deletion or addition line material. We have used this protocol to identify a sequence which has been deleted in both the tetraploid and hexaploid wheat mutants for the homoeologous chromosome pairing locus.


Retroviral vectors containing putative internal ribosome entry sites: development of a polycistronic gene transfer system and applications to human gene therapy. Recombinant retroviral vectors producing multicistronic mRNAs were constructed. Picornavirus putative internal ribosome entry sites (IRES) were used to confer cap-independent translation of an internal cistron. Internal cistrons were engineered by ligation of various lengths of the IRES of encephalomyocarditis (EMC) virus or polio virus to the E. coli chloramphenicol acetyltransferase (CAT) gene. The IRES/CAT fusions were introduced into retroviral vectors 3' to the translation stop codon of the neomycin phosphotransferase (NEO) gene, and the molecular constructs transfected into retroviral vector packaging lines. Retroviral vector producer cells efficiently express the internal CAT gene product only when the full length IRES is used. Both the EMC/CAT and polio/CAT retroviral vectors produced high titer vector supernatant capable of productive transduction of target cells. To test the generality of this gene transfer system, a retroviral vector containing an IRES fusion to the human adenosine deaminase (ADA) gene was constructed. Producer cell supernatant was used to transduce NIH/3T3 cells, and transduced cells were shown to express NEO, and ADA. Novel three-gene-containing retroviral vectors were constructed by introducing the EMC/ADA fusion into either an existing internal-promoter-containing vector, or a polio/CAT bicistronic vector. Producer cell clones of the three-gene vectors synthesize all three gene products, were of high titer, and could productively transduce NIH/3T3 cells. By utilizing cap-independent translation units, IRES vectors can produce polycistronic mRNAs which enhance the ability of retroviral-mediated gene transfer to engineer cells to produce multiple foreign proteins.


Regulation of the Dictyostelium cAMP-induced, prestalk-specific DdrasD gene: identification of cis-acting elements. We have shown previously that expression of the Dictyostelium ras gene DdrasD (previously denoted Ddras) is induced during multicellular development and in single-cell shaking culture in response to cAMP (1). Analysis of transformants carrying DdrasD/lacZ reporter constructs showed DdrasD expression to be prestalk-specific (2). The gene is transcribed from three start sites with transcription from the distal site producing an approximately 1.2 kb transcript, which is expressed at low levels in growing cells and is subsequently induced late in aggregation. This promoter is also induced to high levels by cAMP. Transcription from the two more proximal sites is coregulated and is induced during development, resulting in approximately 1.0 kb transcripts. In this study, we examine cis-acting regions required for proper regulation of DdrasD expression using a DdrasD/beta-glucuronidase reporter gene construct. We have identified distinct sequence elements required for developmental and vegetative expression of DdrasD. A domain containing a CA repeat, similar to ones found in other late, cAMP-induced Dictyostelium genes, is required for cAMP-induced and developmental expression.


Statistical analyses of counts and distributions of restriction sites in DNA sequences. Counts and spacings of all 4- and 6-bp palindromes in DNA sequences from a broad range of organisms were investigated. Both 4- and 6-bp average palindrome counts were significantly low in all bacteriophages except one, probably as a means of avoiding restriction enzyme cleavage. The exception, T4 of normal 4- and 6-palindrome counts, putatively derives protection from modification of cytosine to hydroxymethylcytosine plus glycosylation. The counts and distributions of 4-bp and of 6-bp restriction sites in bacterial species are variable. Bacterial cells with multiple restriction systems for 4-bp or 6-bp target specificities are low in aggregate 4- or 6-bp palindrome counts/kb, respectively, but bacterial cells lacking exact 4-cutter enzymes generally show normal or high counts of 4-bp palindromes when compared with random control sequences of comparable nucleotide frequencies. For example, E. coli, apparently without an exact 4-bp target restriction endonuclease (see text), contains normal aggregate 4-palindrome counts/kb, while B. subtilis, which abounds with 4-bp restriction systems, shows a significant under-representation of 4-palindrome counts. Both E. coli and B. subtilis have many 6-bp restriction enzymes and concomitantly diminished aggregate 6-palindrome counts/kb. Eukaryote, viral, and organelle sequences generally have aggregate 4- and 6-palindromic counts/kb in the normal range. Interpretations of these results are given in terms of restriction/methylation regimes, recombination and transcription processes, and possible structural and regulatory roles of 4- and 6-bp palindromes.


Purification and characterization of an endo-exonuclease from adult flies of Drosophila melanogaster. An endo-exonuclease (designated nuclease III) has been purified to near homogeneity from adult flies of Drosophila melanogaster. The enzyme degrades single- and double-stranded DNA and RNA. It has a sedimentation co-efficient of 3.1S and a strokes radius of 27A The native form of the purified enzyme appears to be a monomer of 33,600 dalton. It has a pH optimum of 7-8.5 and requires Mg2+ or Mn2+ but not Ca2+ or Co2+ for its activity. The enzyme activity on double-stranded DNA was inhibited 50% by 30 mM NaCl, while its activity on single-stranded DNA required 100 mM NaCl for 50% inhibition. Under the latter conditions, its activity on double-stranded DNA was inhibited approximately 98%. The enzyme degrades DNA to complete acid soluble products which are a mixture of mono- and oligonucleotides with 5'-P and 3'-OH termini. Supercoiled DNA was converted by the enzyme to nicked and subsequently to linear forms in a stepwise fashion under the condition in which the enzyme works optimally on single-stranded DNA. The amino acid composition and amino acid sequencing of tryptic peptides from purified nuclease III is also reported.


Cis and trans regulatory elements required for regulation of the CHO1 gene of Saccharomyces cerevisiae. A 34 base-pair (bp) fragment spanning sequences -154 to -120 of the promoter of the CHO1 gene (structural gene for phosphatidylserine synthase) from the yeast Saccharomyces cerevisiae has been shown to place transcription of a promoter-less Escherichia coli lacZ gene under control of the phospholipid precursors inositol and choline. Furthermore, in deletion experiments the CHO1 UASINO was localized to sequences between -151 and -123 of the CHO1 promoter. A nine bp sequence was identified in the promoter region of the CHO1 gene that shares an eight out of nine bp match with a sequence (consensus 5' ATGTGAAAT 3') that is repeated a total of 23 times upstream from several coregulated phospholipid biosynthetic genes. This sequence is contained within the -151 to -123 region to which the CHO1 UAS has been localized. The nine bp repeated element is believed to be involved in the control of phospholipid biosynthetic gene transcription in response to changing levels of inositol and choline in the growth medium. This control has been shown to require activities encoded by the products of the three regulatory genes: INO2, INO4, and OPI1. A mutation in any of these regulatory genes results in aberrant CHO1-lacZ gene regulation, and affects regulation of the construct containing the 34 bp (-154 to -120) CHO1 fragment demonstrating that the regulatory signal generated by these genes interacts with the 5' end of the CHO1 gene.


[Results of displacement studies on benzodiazepine receptors (123I-iomazenil and flumazenil)]. SPECT studies with 123I-iomazenil were performed in 14 patients with hepatic encephalopathy. The activity concentration in the brain decreases spontaneously by 10 to 20% within 80 min, in the nasal tissue by 35%. After an i.v. injection of 0.1 to 0.5 mg flumazenil a decrease of 29 to 38% is observed within 20 min, with a comparable decrease in white matter and nasal tissue. The application of well-known models of receptor kinetics is therefore impossible.


A quantitative index derived from 99mTc-pertechnetate scintigraphy to assist in the diagnosis of primary Sjögren's syndrome. A safe and simple technique is reported by which primary Sjögren's syndrome can be detected with a relatively high specificity and sensitivity. The method of serial scintigraphy has been used with reasonable success; however, the application of the linearity index as described here produced superior results. In 71 patients investigated, a sensitivity of 87% and specificity of 93% were recorded and make this the method of choice for evaluating patients suspected of having primary Sjögren's syndrome.


The effects of heat, water, acid, and alkali treatment of tomato cannery wastes on growth, metabolizable energy value, and nitrogen utilization of broiler chicks. Two experiments were conducted to determine the effects of heat, water, acid, and alkali treatment of tomato pomace on gain, feed to gain ratio, nitrogen utilization, and ME of diets for broiler chicks. In Experiment 1, both treated and untreated tomato pomace was included in broiler diets at a 10 or 20% level. Results indicated that the level or antinutritional factors present in untreated tomato cannery waste did not appreciably depress any measured production parameter. Hence, it appeared that untreated tomato cannery wastes might be used as a feed ingredient in low-energy poultry diets (broiler breeder and laying hen recycling rations), ruminant diets, and as a protein source in regions of the world where such feed ingredients are scarce. The second experiment was designed to test the effect of alkali concentration and treatment time of tomato pomace on the performance of broiler chicks. Alkali treatment of tomato cannery wastes increased gain and decreased feed to gain ratios of broiler chicks over those of untreated tomato waste controls. Results indicated that the increased gain and decreased feed to gain ratios of the chicks were due in part to the acid neutralization phase of the alkali treatment. Alkali treatment apparently affects the tomato cannery wastes almost instantaneously, as differences among actual treatment times and concentrations were small. However, only the highest alkali treatment increased the pH of the tomato cannery waste above 7, suggesting that a true alkali treatment might cause additional improvements in gain and feed to gain ratio when fed to broiler chicks.


Role of leukotriene B4 in the pathogenesis of hepatic ischemia-reperfusion injury in the rat. A common feature to most models of ischemia-reperfusion injury is the accumulation of polymorphonuclear leukocytes (PMNs) into the post-ischemic tissue during the reperfusion period. Interventions that lead to decreased PMN infiltration protect against tissue injury and therefore a knowledge of the chemotactic mediators leading to PMN accumulation is essential to understanding the pathogenesis of the injury and to the development of successful therapeutic strategies. Leukotriene B4 (LTB4), a metabolite formed via the 5-lipoxygenase pathway from arachidonic acid, is one of the most potent chemotactic mediators known. We have investigated the formation of LTB4 in a well characterized model of hepatic ischemia-reperfusion injury in the rat and made use of a specific leukotriene biosynthesis inhibitor, L663,536, to determine the importance of LTB4 in the pathogenesis of the injury. LTB4 concentrations were measured with a specific and sensitive gas chromatographic-mass spectrometric method previously developed in our laboratory. In liver tissue LTB4 levels were below the detection limit of 20 pg/g before 45 min ischemia and did not increase during the first 6 h of reperfusion. However, at 15 h and 24 h reperfusion LTB4 concentrations had increased to levels 50-fold those in control liver (867 +/- 267 pg/g). The increase of plasma alanine aminotransferase (ALT) activities indicated two phases of injury, an initial phase during the first few hours of reperfusion, and a second more severe injury phase between 6 h and 24 h reperfusion. PMNs accumulated in tissue throughout the reflow period reaching 700 +/- 49 per 50 high power fields (HPF) at 24 h.(ABSTRACT TRUNCATED AT 250 WORDS)


Evaluation of a leukotriene receptor antagonist in prevention of hyperoxic lung injury in newborn rabbits. Prolonged exposure to hyperoxia can result in significant lung injury and has been associated with the development of bronchopulmonary dysplasia. Leukotrienes (LT) recruit polymorphonuclear leukocytes (PMN) to the lung, increase vascular permeability, and have therefore been postulated to play a role in the pathogenesis of hyperoxic lung injury. This study investigates ICI 198,615 (ICI), an LTD4 and LTE4 receptor antagonist in preventing hyperoxic lung injury in newborn rabbits. Matched littermates of 7-day-old rabbits received ICI (0.1 or 1.0 microM/kg/h) or vehicle alone, were exposed to greater than 95% O2, and sacrificed after 48, 72, 84 and 96 h of exposure. Bronchoalveolar alveolar lavage fluid (BAL) of the left lung was analyzed for white cell count, differential, absolute number of PMNs, total protein, and cyclooxygenase products 6-keto-PGF1 alpha, and thromboxane B2. Lung water was quantified utilizing the right lung. Results demonstrated no significant differences between the ICI groups or between the ICI groups and controls. In conclusion, the administration of the LTD4 and LTE4 receptor antagonist ICI 198,615 was insufficient to reduce the formation of pulmonary edema, reduce mortality or attenuate hyperoxic lung injury. These experiments suggest that a number of other mediators may be involved in the hyperoxic lung injury process and that the functional inhibition of a portion of the arachidonic acid cascade was not sufficient to either prevent or attenuate hyperoxic lung injury in newborn rabbits.


Pseudo-meconium ileus due to cytomegalovirus infection: a report of three cases. We observed three cases of antenatal ileus associated with cytomegalovirus (CMV) infection of the fetus and placenta. Two were detected antenatally because of increased echogenicity of the lower abdomen. In the first fetus, the ileus was associated with abnormalities of amniotic fluid enzymes but it was transient and not present at autopsy and the CMV infection was mild, without inflammatory infiltration or necrosis. In the two others, the ileus persisted and CMV-associated lesions were severe. In all three cases the virus was demonstrable in ganglion cells or within myenteric and submucosal plexuses all along the small and large intestine; ileus was imputed to CMV, which caused a paralytic ileus, and in one fetus meconium ileus was also present. A transient episode of ileus does not indicate that the fetus is free of disease and a wide range of causes must be considered, including CMV infection as well as the more usual causes such as cystic fibrosis (CF) and Hirschsprung's disease.


Photosensitization by anticancer agents--10. ortho-semiquinone and superoxide radicals produced during anthrapyrazole-sensitized oxidation of catechols. Photosensitized oxidation of catechol, 3,4-dihydroxybenzoic acid (DHBA), 3,4-dihydroxy-dihydrocinnamic acid (DHCA), and 3,4-dihydroxy-phenylalanine (DOPA) by novel anticancer agents, anthrapyrazoles (AP), has been studied employing EPR and the spin trapping technique. The formation of o-semiquinone radicals, the one-electron oxidation products of the catechols, stabilized in the form of zinc ion complexes, has been demonstrated. Rate constants for the disproportionation of the semiquinone radical/Zn2+ complexes in (DMSO)/acetate buffer (pH 4.5, 1:1 vol/vol; 100 mM Zn2+) mixture have been determined to be 0.35 x 10(4), 14 x 10(4), 8.8 x 10(4) and 3 x 10(4) M-1 s-1 for catechol, DHBA, DHCA and DOPA respectively. The presence of oxygen enhanced rather than inhibited the photogeneration of the o-semiquinone radicals and facilitated their EPR detection. The EPR spectrum of the superoxide radical adduct with the spin trap 5,5-dimethyl-1-pyrroline-N-oxide was observed for the first time during photosensitized oxidation of the catechols in acidic aqueous solutions and in DMSO/acetate buffer mixture.


Enhanced healing of cutaneous wounds in rats using beads with positively charged surfaces. The efficacy of electrical fields in soft-tissue repair is unclear. Materials with a charged surface provide a localized charged environment. We examined the effects of surface-charged particles in wound healing in rats with paired dorsal incisions with one side serving as a control. Tensiometry demonstrated that after 10 days, wounds with positively charged particles were 53 percent stronger (p less than 0.001) than controls (10 rats, 30 wound strips), whereas differences with negatively charged (6 rats, 15 strips) or uncharged beads (11 rats, 33 strips) were insignificant. Histologically, wounds with positively charged particles were characterized by large quantities of collagen-rich connective tissue and by prominent bead-associated giant cells. At 94 days, no differences in wound strength were noted. This method of creating charged local environments has potential clinical implications and may add insights into the behavior of cells in response to charged stimuli.


Quantitative analysis of lidocaine hydrochloride delivery by diffusion across tissue expander membranes. Permeability of Silastic tissue expander shells to lidocaine was studied to investigate the feasibility of intraluminal lidocaine injection for pain relief during soft-tissue expansion. Both intact expanders and an apparatus using isolated Silastic membrane segments were used to partition solutions of various lidocaine concentrations, and the rate of diffusion was quantitatively measured using a fluorescence polarization immunoassay. Lidocaine flux was found to follow Fick's law of passive diffusion with respect to time, surface area, and concentration gradient for the first 9 hours, with a permeability coefficient of 10.3 +/- 2.6 micrograms (h.cm2.percent)-1 (mean +/- SD) and diffusion coefficient of 7.5 x 10(-7) cm2/min for an average membrane thickness of 473 +/- 23 microns. After 9 hours, the lidocaine flux decreased exponentially, although the concentration gradient across the membrane remained essentially the same order of magnitude. Plasma proteins in the outer bathing solution and methylparaben used as a preservative in the standard lidocaine formulation had no influence on the change in transport flux with time. At the end of the linear portion of the diffusion curve, less than 2 percent of the total intraluminal lidocaine had crossed the membrane. Potential toxicity in the event of implant rupture limits the maximum total lidocaine dose to approximately 500 mg within an expander at any one time. Within these limits, the capacity for lidocaine delivery of 500 mg lidocaine by a 640-cc tissue expander would be only 6 mg during the first 9 hours after administration.(ABSTRACT TRUNCATED AT 250 WORDS)


Silicone gel infiltration of a peripheral nerve and constrictive neuropathy following rupture of a breast prosthesis. Following rupture of a subpectoral breast prosthesis, massive amounts of silicone gel migrated into the arm of a patient. The patient developed painful paresthesias and decreased sensation in the cutaneous distribution of the superficial radial nerve. Nerve conduction studies showed both an increase in distal latency and decreased amplitude in this nerve compared with the normal opposite side. Subsequent neurolysis confirmed dense fibrosis surrounding the nerve. Silicone droplets also were observed within the thickened epineurium of the median nerve, but no electrophysiologic evidence of neuropathy occurred. Multiple debridements of the subcutaneous tissue of the arm were necessary. In one of these specimens, histologic sections demonstrated silicone gel infiltration of a subcutaneous nerve. This is the first reported case of silicone gel infiltration of a nerve and constrictive neuropathy associated with a prosthesis rupture.


Imipramine effect on hypothalamic-pituitary-adrenal axis response to hypoglycemia. Six control subjects underwent an insulin tolerance test before and after the administration of therapeutic doses of imipramine hydrochloride for 10 days to investigate effects of tricyclic antidepressants on hypothalamic-pituitary-adrenal axis response to hypoglycemia. The mean steady-state tricyclic blood level was 141 (SD = 66) ng/ml. Baseline levels of glucose, cortisol, and adrenocorticotropic hormone (ACTH) were not affected by the administration of imipramine. After administration of imipramine for 10 days, subjects uniformly had a significantly lower glucose nadir than before its administration (before imipramine: mean = 32 mg/dl; SD = 5; after imipramine: mean = 24 mg/dl; SD = 6). There was no difference in ACTH or cortisol response before and after the administration of imipramine. These findings suggest that imipramine hydrochloride increases sensitivity to the hypoglycemic effects of insulin, but does not alter the counterregulatory response of ACTH and cortisol.


Free radical yields in A:T polydeoxynucleotides, oligodeoxynucleotides, and monodeoxynucleotides at 4 K. Dose-response curves for free radical trapping in monomers, dimers, and polymers of dAMP and TMP have been measured. Powder samples pressed into pellets were X-irradiated and observed at 4 K using EPR at Q-band microwave frequencies. The initial free-radical yield (G value) and the destruction rate (k value) are reported for 22 samples. The absolute magnitude and relative changes in G are informative. For example, the duplex of homopolymers, poly(dA):poly(T)(Na), gives a large G of 6 while the closely related duplex of alternating ATs, poly(dA-T)(Na), gives a G value of 3. At G of around 6, the range of the bound electrons (e-) and electron loss centers (holes) is believed to be very limited. Two factors are suggested as important to limiting the range, proton transfer between strands and the low probability of radical transfer between strands. The lower G values are viewed as being due to relatively small increases in the range of e- and/or holes.


Depression of an instrumental response by a single devaluation of its outcome. In three experiments rats were trained to make two different instrumental responses for different outcomes. Then the outcomes were given a single differential conditioning experience with lithium chloride. That differential treatment was sufficient to produce differential instrumental responding in a subsequent test. Re-exposure to the devalued outcome was not necessary for an impact on instrumental performance.


The effect of calcitonin-gene-related peptide on acute ischemia-reperfusion renal injury: ultrastructural and membrane lipid peroxidation studies. It is well accepted that postischemic reperfusion promotes functional and morphological impairment which may be related to oxygen free-radical-mediated membrane damage. A new purified bioactive compound, calcitonin-gene-related peptide (CGRP), is known to be not only a potent vasodilator but also a cytoprotective agent. This study was designed to observe whether CGRP has a protective effect on the ischemic kidney. Male Sprague-Dawley rats were subjected to a 45-min period of renal ischemia followed by 60 min of reperfusion. At the beginning of the reperfusion, 12 rats were given intravenous saline and served as controls whereas 5 rats were given CGRP, 10 micrograms/kg intravenously. After reperfusion the kidneys were removed for light- and electronmicroscopy, and the lipid peroxidation product malonaldehyde (MDA) was assayed by thiobarbituric acid (TBA) colorimetry. The results demonstrated that the serum creatinine (Scr) and renal MDA content in the CGRP group were significantly lower than those in the control group. The mean values for Scr were 0.75 +/- 0.09 vs 0.93 +/- 0.05 mg/dL or 62.8 +/- 9.7 vs 82.2 +/- 4.4 mumol/L (p less than 0.05), respectively; while the mean values for MDA were 18.71 +/- 2.13 vs 30.32 +/- 1.78 nmol/100 mg (ww) (p greater than 0.05), respectively. The same signals of free radicals in the ischemic-reperfused kidney with or without CGRP were found by electron spin resonance. Morphological studies demonstrated that the treatment with CGRP ameliorated the ischemic-reperfusion injury to both renal brush borders and mitochondria. The results showed that CGRP has a protective action on ischemia-reperfusion renal injury by decreasing lipid peroxidation of membranes and suggest that it may be a beneficial agent for therapy of acute renal failure.


Malignant histiocytosis in childhood: a distinctive CD30-positive clinicopathological entity associated with a chromosomal translocation involving 5q35. The clinicopathological data on 20 cases of malignant histiocytosis (MH) collected over a period of 30 years at the Hôpital des Enfants Malades (Paris) are reported. Childhood MH was characterized by disseminated, frequently tender lymphadenopathy (19/20), skin (8/20), bone (6/20), and soft tissue localizations (7/20). These features were usually accompanied by fever, deterioration of general condition, and hematological abnormalities including anemia, thrombocytopenia, and occasionally fibrinopenia. These manifestations were clinically suggestive of a diagnosis of a severe neoplastic blood disease, although this hypothesis was not entertained for a long time because of the initial absence of abnormal cells in the blood and bone marrow. MH was characterized by the proliferation of large "histiocyte-like," usually mononucleated cells. When suitable material was available, MH cells appeared to react positively with acid-phosphatase, alpha-naphthyl acetate esterase (ANAE), alpha-antichymotrypsin, and antibodies directed against EMA, HLA DR, CD25, CD30, CD68, and CD71. No B- and T-cell antigens (except for one case) have been detected. Due to the frequent abundance of accompanying granulocytes, lymphoid, and plasma cells, and the presence of areas of necrosis, an initial correct diagnosis of MH was often difficult to establish on skin (four cases), bone (two), and soft tissue (three) biopsies. In lymph nodes, the sinusoidal and perifollicular topography of cell proliferation represented a highly reliable morphological feature. A permanent cell line (DEL) was obtained from a pleural effusion showing a t(5;6)(q35;p21) translocation and a monoallelic immunoglobulin (IgjH) rearrangement and consistent levels of expression of c-fms, c-myc, c-myb, c-ki-ras and c-fgr. Since an identical 5q35 breakpoint has been reported in four other MH cell lines with a comparable phenotype and in several isolated published cases, this chromosomal abnormality provides a highly valuable argument for individualizing an authentic malignancy of the mononuclear phagocyte system (MPS) in childhood, among the rather heterogeneous group of the CD30+ anaplastic large cell lymphomas.


Role of excitation-contraction coupling in muscle fatigue. The force produced by muscles declines during prolonged activity and this decline arises largely from processes within the muscle. At a cellular level the reduced force could be caused by: (a) reduced intracellular calcium release during activity; (b) reduced sensitivity of the myofilaments to calcium; or (c) reduced maximal force development. Experiments involving intracellular calcium measurements in isolated single fibres show that all 3 of the above contribute to the decline of force during fatigue. Metabolic changes associated with fatigue are probably involved in each of the 3 factors. Thus the accumulation of phosphate and protons which occur during fatigue cause a reduction in calcium sensitivity and a decline in maximal force. The cause of the reduced intracellular calcium during contractions in fatigue is less clear. During prolonged tetani the conduction of the action potential in the T-tubules appears to fail leading to reduced intracellular calcium in the central part of the muscle fibre. However, during repeated tetani there is a uniform decline of intracellular calcium across the fibre and this remains one of the least understood processes which contribute to fatigue.


[Comparison of antibody titers against Aujeszky's disease virus in pig blood collected on the farm and in the slaughterhouse]. Antibody titres against Aujeszky's disease virus (ADV) in the plasma of slaughter pigs were compared with respect to the site of sampling, namely on the farm (one day before delivery to the slaughterhouse) and in the slaughterhouse during slaughter. No significant differences were noted in the antibody titres against ADV with respect to the different sampling sites. Blood samples collected at the slaughterhouse can be used for monitoring and surveillance systems within the framework of organised pig health care.


Effects of charge and lipophilicity on mercurial-induced reduction of 45Ca2+ uptake in isolated nerve terminals of the rat. The goal of this study was to compare the ability of neurotoxic mercurials which differ in ionic charge and/or lipophilicity to block nerve-terminal calcium channels. To do so, we examined the acute effects of methyl mercury (MeHg+), ethyl mercury (EtHg+), inorganic mercury (Hg2+), dimethyl mercury (Me2Hg), p-chloromercuribenzoate (PCMB), and p-chloromercuriphenyl-sulfonate (PCMBS-) (10-1000 microM) on 45Ca2+ flux into rat forebrain synaptosomes at rest and during depolarization. Basal (depolarization-independent) entry of 45Ca2+ was measured during 10-sec exposure to mercurials in solutions containing 5 mM KCl. Concentrations of 50, 100, 250, 500, and 1000 microM of Hg2+, MeHg+, and EtHg+ reduced basal influx of 45Ca2+. PCMB reduced basal influx at concentrations of 10, 50, and 100 microM, but increased influx at 1000 microM. PCMBS- and (Me)2Hg had no effect on basal flux at any concentration tested. Uptake of 45Ca2+ was measured after 1 sec of K(+)-induced depolarization (41.25 mM) to determine influx through voltage-dependent Ca2+ channels ("fast" phase) or during the last 10 sec of a 20-sec period of depolarization for uptake associated with a reversed Na+/Ca2+ exchanger and a residual noninactivating Ca2+ channel component ("slow" phase). Fast and slow components of 45Ca2+ uptake into synaptosomes were blocked in a concentration-dependent manner by MeHg+, EtHg+, and Hg2+. For block of the fast component, the calculated IC50's and confidence intervals were (microM) EtHg+, 92 (82, 102); Hg2+, 155 (149, 161); and MeHg+, 196 (120, 272). IC50's and the confidence intervals for the slow component of uptake were (microM) Hg2+, 49 (43, 55); MeHg+, 72 (67, 77); and EtHg+, 147 (142, 152). In contrast, Me2Hg, PCMB, and PCMBS- (10-1000 microM) caused no appreciable reduction in either phase of 45Ca2+ uptake. Increasing [Ca2+]e was unable to overcome the block induced by MeHg+ and EtHg+ (100 microM) on either phase of 45Ca2+ uptake into synaptosomes. Likewise, increasing [Ca2+]e failed to overcome block of the slow component by Hg2+ (100 microM). Increasing [Ca2+]e was able to overcome, in part, block of the fast phase induced by Hg2+ (100 microM) although the percentage of reversal was not statistically significant. The magnitude of block of 45Ca2+ uptake increased as a function of increasing [K+]e for MeHg+ and EtHg+, suggesting the block to be voltage-dependent. Thus, mercurials of dissimilar charge and lipophilicity affect synaptosomal Ca2+ uptake differentially.(ABSTRACT TRUNCATED AT 400 WORDS)


Quantitative immunoassay of total cellular GAP junction protein connexin32 during liver regeneration using antibodies specific to the COOH-terminus. A radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) were used to determine relative concentrations of liver connexin32 (CX32) in rats. The RIA and ELISA utilize synthetic peptides corresponding to regions of the carboxyl-terminus and antibodies raised in rabbits against these peptides. Assuming that affinities of antisera are similar for peptide and native CX32, total cellular CX32 was found to exceed the amount of gap junction protein at the cell surface calculated from morphometric analyses by 1.5-2.0 fold. This finding raises the possibility that some of the protein is present in cytoplasmic compartments or as occult precursors in the plasma membrane. Studies of CX32 content in regenerating rat liver support this conclusion and show a time course of loss and recovery of CX32 that agrees with those reported in studies using other techniques.


6-Methyl-1,3,8-trichlorodibenzofuran (MCDF) as an antiestrogen in human and rodent cancer cell lines: evidence for the role of the Ah receptor. 6-Methyl-1,3,8-trichlorodibenzofuran (MCDF) is a relatively nontoxic analog of 2,3,7,8-tetrachlorodibenzo-p-dioxin. Treatment of aryl hydrocarbon (Ah)-responsive MCF-7 human breast cancer cells with 100 nM MCDF resulted in the inhibition of 17 beta-estradiol-induced proliferation and the secretion of the 34-, 52-, and 160-kDa proteins. After treatment of the cells with 17 beta-[3H]estradiol, 100 nM of MCDF caused a decrease in the accumulation of the radiolabeled nuclear estrogen receptor (ER) complex in these cells. In parallel experiments, the antiestrogenic effects of MCDF were also determined in Ah-responsive wild-type Hepa 1c1c7 cells and Ah-nonresponsive class 1 and class 2 mutant cells. Treatment of the wild-type cells with 17 beta-[3H]estradiol and 100 nM MCDF caused a decrease in the accumulation of radiolabeled nuclear ER complex in these cells whereas no significant effects were observed in the mutant cells as determined by velocity sedimentation analysis. Comparable results were obtained using ER antibodies to measure the decrease in immunoreactive nuclear ER. In addition, both actinomycin D and cycloheximide inhibited the MCDF-mediated decrease of nuclear ER levels in the Hepa 1c1c7 wild-type cells. Although 100 nM MCDF did not induce cytochrome P-450-dependent monooxygenases in the MCF-7 or Hepa 1c1c7 cell lines, incubation of nuclear extracts from the MCF-7 cells treated with 100 nM MCDF with a synthetic consensus dioxin responsive element (an oligonucleotide duplex of 26 bases) gave a retarded band in a gel-retardation assay. The data suggest that the antiestrogenic effects of MCDF does not require the induction of the CYP1A1 gene expression but may involve the induction of other genes.


Changes of neuronal transmission in the hippocampus after transient ischemia in spontaneously hypertensive rats and the protective effects of MK-801. I studied the mechanism of postischemic neuronal degeneration in the hippocampus by an electrophysiological method. Sequential changes of field potentials evoked by perforant path stimulation in the dentate gyrus and the CA1 region of the hippocampus were evaluated in spontaneously hypertensive rats up to 7 days after transient global ischemia induced by bilateral occlusion of the carotid arteries for 20 minutes after electrocauterization of the vertebral arteries. Animals were treated with vehicle or the excitotoxin antagonist (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10 amine (MK-801, 2 mg/kg or 5 mg/kg) intraperitoneally 30 minutes before ischemia. Complete recovery of the population spike was observed in the dentate gyrus within 24 hours after recirculation, followed by a gradual reduction of population spike amplitude. In contrast, population spike in the CA1 region showed partial recovery 24 hours after recirculation, and an abrupt reduction of population spike amplitude occurred on day 2. There was no significant enhancement of population spike amplitude in either region throughout the experiment. Interneuronal recurrent inhibition in the dentate gyrus was enhanced on day 4, and ischemic changes were apparent in the CA1 pyramidal cells on day 7. Pretreatment with 5 mg/kg MK-801 prevented field potential and pathological changes completely in the dentate gyrus and partially in the CA1 region. My results indicate that pathological changes of the CA1 pyramidal neurons after transient ischemia may not be the result of postischemic overstimulation. However, neuronal transmission in the CA1 region may be persistently impaired during or after transient ischemia.


The thymic factor tactivin prevents ACTH from stimulating steroidogenesis by mouse adrenal cells. The effect of tactivin, a thymic hormonal factor, (MW 1-6 KD), on the adrenal glucocorticoid function in mice was studied. Tactivin (0.1-2 micrograms/mouse i.p.) produced a slight decrease in plasma corticosterone. The decrease was much more pronounced when tactivin was administered to mice with a high basal level of the hormone. The tactivin supplement had a significant suppressive effect on corticosterone production in both the whole adrenals and the intact isolated adrenal cells. When added at the doses of 0.00064-2 micrograms/ml to the isolated adrenal cells in the presence of ACTH, tactivin abolished the stimulatory effect of corticotropin on corticosterone production. The abolition was complete at a low dose (1.6 microIU/ml) and incomplete, yet significant, at a high dose of ACTH (1600 microIU/ml). The in vitro data are in a good agreement with those obtained in the in vivo experiments. The stimulatory effect of a synthetic analog of cAMP-Dibutyryl-cAMP on the steroidogenesis of adrenal cells was as pronounced as that of ACTH. Nevertheless, tactivin exerted no influence on Bu2-cAMP stimulation. The results indicate that tactivin prevents ACTH from acting on steroidogenesis at some time point preceding the formation of secondary messengers.


Human immunodeficiency virus, human papillomavirus, and cervical intraepithelial neoplasia in Nairobi prostitutes. To evaluate the impact of human immunodeficiency virus (HIV) on human papillomavirus (HPV) and cervical intraepithelial neoplasia (CIN), a study was conducted of 147 HIV-seropositive and 51 HIV-seronegative prostitutes in Nairobi, Kenya. Among the women infected with HIV, 10 (7%) had signs or symptoms of significant HIV-related disease, and the remaining 93% were asymptomatic. The prevalence of cervical HPV DNA was 37% among HIV-seropositive women and 24% in HIV-seronegative women (odds ratio [OR] 1.7, 95% confidence intervals [CI] 0.8, 3.6, after adjusting for potential confounding factors). Genital warts, cervical HPV DNA, and cytologic findings consistent with CIN were all significantly associated with younger age and fewer years of prostitution, but were unrelated or weakly related to number of sexual partners per week or frequency of condom use. In a subset of 63 women with evaluable Papanicolaou smears, CIN was found in 50% of the women with HPV but only in 8% of those without HPV (adjusted OR 7.2, 95% CI 1.6, 32.1, P = 0.006). However, CIN was unrelated to HIV seropositivity (prevalence of 26% among HIV-seropositive women and 24% in HIV-seronegative women). Among women with cervical HPV DNA, HIV infection was not associated with an increased prevalence of CIN (47% prevalence among women with HIV versus 57% prevalence among women without HIV). Thus, in this population of HIV-seropositive women, most of whom had CDC Stage II or III infection, there was no demonstration of an adverse impact of HIV on CIN.


Suppressive effect of tetrahydrocannabinol on specific T cell subpopulations in the thymus. The effects of THC (delta-9-tetrahydrocannabinol) on thymocytes from adult BALB/c mice were investigated following in vitro stimulation with the T cell mitogen Concanavalin A (Con A). Con A treatment resulted in a decrease in the relative percent of L3T4+/Ly2+ cells (double positive; immature) with a concomitant increase in the percent of Ly2 cells (suppressor/cytotoxic). No change in the percent of L3T4 cells (helper) occurred. In addition, Con A stimulation resulted in an increase in the absolute number of Ly2 cells, with no change in number of L3T4 cells. Stimulation with the mitogen led to an increase in fluorescence intensity (indicative of relative number of markers) of Ly2 markers on Ly2 single positive cells as well as on L3T4+/Ly2+ cells. There was a moderate mitogen induced increase of L3T4 markers, but only on L3T4 single positive cells. Exposure to THC resulted in a definitive suppressive effect on Con A induced thymus cell proliferation. The suppression was primarily evidenced in the single positive Ly2 subpopulations. Since the thymus is the source of immunocompetent lymphocytes to the secondary lymphoid organs, it seems plausible that marijuana negatively affects maturation of these cells in the developing individual.


Multiple benzodiazepine receptors: no reason for anxiety. Since the introduction of the benzodiazepines into clinical practice in 1960, these drugs have been widely employed as anxiolytics, sedative/hypnotics and anticonvulsants. In recent years, concern has been expressed about their side-effects, and their use has declined. During this latter period many advances have been made in understanding the molecular mechanisms by which these drugs produce their effects. Adam Doble and Ian Martin review this progress and highlight the possibilities that these advances may hold for the development of more efficacious and specific medicines.


[The synchronous appearance of multiple testicular epidermoid cysts and of a malignant mixed germinal tumor in the contralateral testis]. We describe the simultaneous appearance of multiple epidermoid cysts in the right and an adult teratoma containing embryonal carcinoma and choriocarcinoma in the left testis. No similar case has previously been described. Epidermoid cysts of the testis are rare, accounting for about 1% of all testicular tumors. Epidermoid cysts are now regarded as monoepidermally developed teratomas of germ cell origin. Testicular teratomas in adults, however, are always malignant. Because epidermoid cysts are rare tumors, primary therapy often consists in ablation of the testis. In the case described, excision of the epidermoid cyst protected the patient from complete castration. This case demonstrates the simultaneous appearance of a malignant and a benign testicular germ cell tumor.


[Results of detection of hepatitis C virus antibodies (anti-HCV) in blood donors and selected patients]. The authors examined 509 donors for potential carriership of the virus of hepatitis C (HCV), using kits for the detection of anti-HCV. Concurrently they examined also 110 hameatological patients who had a major transfusion of blood, plasma and its derivatives. They found that in the so-called healthy population anti-HCV is present in 0.39%. In haematological patients, i.e. patients with frequent haemotherapy they detected 29 positive cases, i.e. 26.36%. The largest number of anti-HCV positive patients was in the group of haemocoagulopathies. The investigation confirmed the close association between haemotherapy and the transmission of HCV. The authors draw attention to the need to introduce anti-HCV examinations in the transfusion service as an important preventive provision.


Benign follicular thyroid nodule composed of signet-ring-like cells with PAS-negative thyroglobulin accumulation in dilatated rough endoplasmic reticulums. A benign follicular thyroid nodule composed of signet-ring-like cells in a 46-year-old Japanese euthyroid woman is reported. Thyroglobulin was positive, and periodic acid-Schiff (PAS) and other mucin stainings were negative in the cytoplasm of signet-ring-like cells. Ultrastructurally their cytoplasm contained many dilatated rough endoplasmic reticulums (rERs), the largest of which compressed the nucleus toward the cellular apical portion. Functional disturbance of rER or Golgi apparatus might be the cause of this characteristic cellular change.


Small cell (endocrine cell) carcinoma of the gallbladder with squamous and adenocarcinomatous components. Small cell (endocrine cell) carcinoma of the gallbladder in a 62-year-old woman is reported. The palliative cholecystectomy specimen revealed a submucosally invading tumor with extensive hemorrhagic necrosis. At autopsy, performed five months after surgery, a huge tumor measuring 14 x 12 x 8 cm was located at the liver hilus. No signs or symptoms related to overproduction of hormones were recorded throughout her illness. Neither lung lesions nor gall stones were identified. Histologically, diffuse proliferation of small, spindle-shaped atypical tumor cells with numerous mitoses was evident. Intraepithelial tumor cell proliferation in the gallbladder mucosa was seen focally. The neuroendocrine nature of the tumor cells was confirmed by the histologic pattern of growth with pseudo-rosette formation, positive reaction for Grimelius' argyrophilia, neuron-specific enolase and Leu 7, and ultrastructural demonstration of neuroendocrine-type granules. Immunostaining for a variety of hormones was all negative. Characteristically, foci with squamous and adenocarcinomatous differentiation were identified in the tumor tissue. The glandular components were immunoreactive for carcinoembryonic antigen, secretory component, epithelial membrane antigen and CA19-9. The histogenesis and totipotentiality of the neoplasm were discussed.


Peripheral T-cell lymphoma of AILD (angioimmunoblastic lymphadenopathy with dysproteinemia) type involving gastrointestinal tract. A morphologic, phenotypic and genotypic study. A case of angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) which showed widespread involvement of the gastrointestinal tract is reported. A lymph node biopsy specimen showed the characteristic histological features of AILD. During the progression of the illness, lymphomatous lesions developed in the gastrointestinal tract, complicated by cytomegalovirus infection. A double immunoenzymatic study using a combination of Ki-67 antibody and antibodies against surface antigens demonstrated that CD3+, CD4+, and/or T-cell receptor (TCR) beta+ cells were predominant (67-68%) among the population of proliferating Ki-67% cells, rather than CD8+ or CD22+ cells. Clonal rearrangement of the TCR beta chain gene was also detected. These findings provide further evidence for the neoplastic nature of lesions of this type, and the diagnosis of peripheral T-cell lymphoma.


Intra-pancreatic, extra-tumoral perineural invasion (nex). An indicator for the presence of retroperitoneal neural plexus invasion by pancreas carcinoma. A histopathologic study of 65 resected malignant lesions of the pancreas was done to search for a histologic predictor for the presence of extra-pancreatic neural plexus involvement by carcinoma cells. Extra-pancreatic neural plexus invasion by pancreas carcinoma cells, plx(+), was detected in 28 of 65 cases (43%). plx(+) was not significantly associated with tumor location, tumor size, histologic type, or lymph vessel invasion (ly) by carcinoma cells but was significantly associated with rpe. Plx(+) was also significantly associated with mixed moderate and severe degrees of intra-pancreatic neural invasion (ne2/ne3 group), but not with mixed no and slight degrees (ne0/ne1 group). However, fourteen plx(+) cases were excluded in the ne2/ne3 group; they were found in the ne1 group. Therefore, ne factors appear to have a certain but limited usefulness for the prediction of plx(+). When a perineural invasion by tumor cells was found within the pancreas but outside of the major mass of cancer, it was designated "intra-pancreatic, extra-tumoral perineural invasion (nex)". The presence of nex(+) was found in 37 cases, in which 24 out of the 28 plx(+) cases (85.7%) were included. A statistically significant association was found between plx(+) and nex(+). In particular, 12 out of the 14 plx- positive ne1 cases were nex(+). In ne1 cases, 33.3% (14/42) were plx(+), whereas in ne1 and nex(+) cases, 60% (12/20) were plx(+). There was a statistical significance between these two figures (p less than 0.05). Thus, nex(+) appears to be a useful indicator for plx(+), particularly in ne1 cases.


Suicide and community psychiatric care--a preliminary report. A preliminary survey of 34 suicides among patients attending community services for the chronically mentally ill revealed a rate of 520 per 100,000 admitted. In contrast to earlier hospital surveys, no risk variables were identified for patients dying by suicide. Thirty-four percent of suicides occurred within one week of the last treatment and 59% within 3 months of service entry. It appears that early and intensive follow-up may be necessary to prevent suicide among patients receiving community psychiatric care.


Polyneuropathy caused by chlorprothixene. Seven chronic psychiatric patients developed signs of polyneuropathy mainly in the lower limbs after receiving chlorprothixene (500-1800 mg/d) from 3 to more than 24 months. The most prominent electroneuromyographic findings were decreased or not measurable motor conduction velocities of the peroneal nerves. Electromyography showed signs of denervation in the leg muscles in all the patients, and also in the upper limbs in 4 of them. The signs of polyneuropathy gradually subsided after withdrawal of chlorprothixene. Thus, chlorprothixene may cause a toxic, dose-dependent, reversible polyneuropathy.


[Pyelolymphatic backflow complicating acute hemorrhagic cystitis: a case report]. The clinical course of a case of pyelolymphatic backflow complicated with acute hemorrhagic cystitis is reported. The patient was a 19-year-old woman. She had disturbed passage in the intravesical ureter due to acute hemorrhagic cystitis caused by adenovirus. Pyelolymphatic backflow occurred because of increased pelvic pressure.


Substance use disorders: predictions for the 1990s. Numerous factors will affect the course of alcohol-drug services over the next decade, including technological-scientific advances, legislation, professional training standards, case law, availability and cost of substances, and social policy toward users. Despite some apparent lessening of substance use, high levels of substance abuse are likely to continue due to fetal damage in high-risk offspring, younger onset of substance abuse, and solo-parent families. Care givers must be prepared to treat more youth, elderly, women, minority, and "dual disorder" patients. Federal and local statecraft against substance production and use remains crude and does not show signs of the increasing sophistication observed elsewhere in the world. Although these forces favor continued high levels of substance abuse problems, the funding mechanisms to provide care are under increasing assault by both the private and public sectors. Areas in which professional practice are apt to improve include clinical assessment-reassessment, treatment outcome research, monitoring during recovery, and out-patient treatment. Professional groups and treatment organizations will become more proactive in the financial support and management of treatment services.


Progression of dependence in male cocaine addicts. We studied the self-reported temporal sequence of cocaine-related problems in 45 male predominantly Black (85%), lower SES cocaine addicts undergoing inpatient treatment at a large urban VA psychiatric hospital. Subjects reported recent average use of 2.5 g of cocaine per day for 14 days each month and experiencing a mean of 14 cocaine-related problems. The temporal sequence of cocaine-related problems was roughly consistent with the sequence of alcohol-related problems reported for alcoholics, with the earliest problems being interpersonal (e.g., arguments with others) and the most recent problems the severest (e.g., job loss, marital separation). The cocaine addicts showed a faster progression from first cocaine use to first cocaine-related problems (mean of 3.75 years) than that reported for alcoholics from first drink to first heavy drinking (8-10 years). Cocaine smokers had a faster course (3.4 years) than intranasal users (5.3 years).


Retention of cocaine abusers in outpatient psychotherapy. Retention in outpatient psychotherapy of 148 crack and cocaine abusers was examined. The clients were predominantly Black (63%) and Hispanic (21%), predominantly male (87%), and the majority (66%) had completed no more than 12 years of high school. Clients entered treatment in a low-cost treatment center in New York City between June 1987 and November 1988. Forty-two percent (62) of the subjects were seen for one or two research interviews only, and did not return to begin therapy. Of the 86 persons who came to at least one therapy session, 30% (26) dropped out before the third session, 28% (24) dropped out between the third and fifth sessions, and 42% (36) were retained for six or more sessions. Short-term and longer-term retentions were analyzed separately, using a battery including sociodemographic variables, treatment history, psychiatric symptomatology, number of arrests, and drug use variables. None of the variables considered was significantly related to short-term retention. There were large although not significant differences in longer term retention by therapist. Longer-term retention was associated significantly with being White (contrasted with being Black) and being young. Nonsignificant but large associations were found between longer-term retention and having few arrests, being Hispanic (contrasted with being Black), and having low SCL-90 scores. Results are compared with previous findings about retention in drug and alcohol treatment. It is suggested that future research on retention in treatment focus less on client variables and more on therapist and program variables.


Treatment of crack cocaine use with carbamazepine. Crack is a rock crystalline alkaloid form of cocaine which can be smoked. At the University of Minnesota, we have developed an experimental pharmacologic treatment for cocaine abusers. Of 26 patients treated to date, 16 have been crack cocaine users. During the hundred days preceding treatment, the 16 crack subjects used cocaine by all routes an average of 71 days each. Improvement was based on a self-reported decrease in cocaine frequency of use. Using carbamazepine, seven highly successful and six partially successful patients reduced their use to 0.7 days and 26 days per 100 days, respectively. These results, though hopeful, must be viewed with caution and considered preliminary and tentative.


Cocaine-associated panic attacks in methadone-maintained patients. The incidence of panic attacks methadone-maintained patients has increased over a 10-year period from 1 to 6-13%. Cocaine use appears to be associated with this increase, although other environmental and constitutional factors may be contributory. Patients with cocaine-associated panic differ from other panic patients in rates of psychiatric hospitalization and medical illness, but not in depression, other drug use, or agoraphobia.


Characteristics of drug users admitted to alcohol detoxication centers. A survey of admissions to Ontario's alcohol detoxication centers showed many admissions used alcohol in combination with other drugs, and a poor correspondence between self-reports and results of the urine analysis. The use of THC was more common among younger admissions with low urine alcohol levels. Benzodiazepine use was more common among referrals from hospitals, general practitioners, and those presenting with cuts and bruises.


Serum prolactin level, craving, and early discharge from treatment in cocaine-dependent patients. On admission to inpatient treatment, 13 of 33 (39%) cocaine-dependent patients were found to be hyperprolactinemic. Cocaine craving was significantly correlated with cocaine use during the 30 days prior to admission. Serum prolactin level, however, was not correlated with either cocaine use or craving. This lack of intercorrelation argues against the notion that dopamine depletion is an adequate explanation for both craving and hyperprolactinemia following chronic cocaine use. Hyperprolactinemia was associated with a greater likelihood of early discharge from treatment. Gender differences, recent use of other drugs (including alcohol), and a diagnosis of antisocial personality disorder were not related to early discharge. The clinical relevance of these findings is discussed. Further investigation is needed to determine the etiology of the hyperprolactinemia that has been observed in some cocaine-dependent patient samples.


Treatment compliance after detoxification among highly disadvantaged alcoholics. An outcome study was carried out on a series of 109 highly disadvantaged alcoholics discharged from the detoxification unit of a large municipal hospital in New York City. We examined the impact of a variety of clinical and demographic factors on retention in the initial phases of outpatient and inpatient treatment following discharge. Both high school completion and a history of at least 6 months of employment in the two years preceding admission correlated with frequency of registration for continued aftercare. Measurements of cognitive flexibility correlated with frequency of aftercare completion. An association strongly approaching significance was also found between length of hospital stay and aftercare completion. Some suggestions are made as to the assessment and aftercare planning for highly disadvantaged alcoholics.


Corticotropin-independent effect of ovine corticotropin-releasing hormone on cortisol release in man. The effect of corticotropin-releasing hormone (CRH), independent of adrenocorticotropin hormone (ACTH), was evaluated in nine healthy individuals. Cortisol release and corresponding ACTH production were determined after separate intravenous administration of ovine-CRH (1 micrograms/kg BW) and insulin inducing hypoglycemia (0.1 u/kg BW). Adrenocorticotropin hormone (1-24; 250 micrograms intravenous bolus) revealed an adequate adrenal reserve capacity in all subjects. At the time of peak cortisol response following CRH and insulin administration, IR-cortisol increments were 14 +/- 1 micrograms/dl and 9 +/- 1 micrograms/dl (mean +/- SE), respectively (p less than .05); whereas ACTH (IR-ACTH) increments were 40 +/- 10 ng/l and 53 +/- 14 ng/l, respectively. The cortisol increment/ACTH increment ratios were 0.53 +/- 0.09 and 0/36 +/- 0.09, respectively (p less than 0.05), suggesting an ACTH-independent effect of CRH on cortisol production. The authors speculate that CRH may have a direct effect on the human adrenal gland or it may release ACTH-like factors that stimulate the human adrenal cortex.


Case report: metastatic germ cell tumor presenting with thromboembolism and bilateral pneumothorax. Germ cell tumors usually present with symptoms that are attributed to their location. This article describes a patient who had an extragonadal tumor that was not discovered until he experienced several tumor-related complications.


Presence of Epstein-Barr virus in Hodgkin's disease is not exclusive to Reed-Sternberg cells. Thirty-three cases of Hodgkin's disease (HD) have been studied for the presence of Epstein-Barr virus (EBV) using a novel nonisotopic in situ hybridization procedure, based on the detection of Epstein-Barr encoded RNAs with oligonucleotide probes. An intense and morphologically distinct nuclear staining, sparing the nucleolus was seen in a total of 12 cases (36%). In six of these cases, the signal was located to the Hodgkin and Reed-Sternberg cells (HR-S); in the other six positive cases, the signal was observed only in the non-neoplastic small lymphocytes. These lymphocytes were few in number and immunocytochemistry results were consistent with a B-cell phenotype. The presence of EBV in those cases characterized by nuclear staining of small lymphocytes was confirmed by the polymerase chain reaction (PCR) analysis. The authors report the detection of EBV in small lymphocytes in HD by in situ hybridization and discuss the implications of these findings in relation to the proposed etiologic association between EBV and HD.


Epstein-Barr virus-associated gastric adenocarcinoma. The Epstein-Barr virus (EBV) has been detected in certain types of lymphoma and some epithelial neoplasms including nasopharyngeal lymphoepithelioma, and rare lymphoepithelioma-like carcinomas occurring in a variety of organs including, most recently, the stomach. The authors investigated the possibility that EBV may be present not only in the rare gastric cancers that resemble nasopharyngeal lymphoepithelioma, but also in typical gastric adenocarcinoma. EBV sequences were detected in 22 of 138 (16%) cases of typical gastric adenocarcinoma by polymerase chain reaction and in situ hybridization (ISH) techniques. The EBV genomes were specifically present within the gastric carcinoma cells in an even distribution. The EBV genomes were also present in adjacent dysplastic epithelium but were absent in surrounding lymphocytes, other normal stromal cells, intestinal metaplasia, and normal gastric mucosa. The EBV genomes in the infected gastric carcinoma cells are expressed as EBV RNA was detected by ISH. EBV was most often detected in gastric tumors from men (21%) compared with women (3%). Thus some cases of gastric adenocarcinoma are EBV-associated.


Genomic changes in the WT-gene (WT1) in Wilms' tumors and their correlation with histology. The authors studied genomic changes in unilateral Wilms' tumors by using WT33, a candidate cDNA for the tumor, and their correlation with histology. By Southern blot analysis, three cases of genomic deletions of both alleles were found in 25 tumors. The three tumors that showed genomic deletions were histologically classified as triphasic nephroblastic Wilms' tumor and one of them was associated with intralobar nephroblastomatosis and a rhabdomyomatous component. In one case, the WT1 gene was totally deleted, in another case, the 3' region of the gene was partially deleted, and in the last one, the deletion of DNA was intragenic. This is the first report of a comparison of genomic alteration with histopathology. These findings show new aspects of the role of the WT1 gene in the development of Wilms' tumor.


Filaments of Lewy bodies contain insoluble cytoskeletal elements. The Lewy body is an intraneuronal inclusion body that is one of the histologic hallmarks of Parkinson's disease, a degenerative disease of the brain. Ultrastructural analysis has shown that the Lewy body is composed of straight 7-20 nm filaments and amorphous elements. Previous light microscopic, immunocytochemical studies have suggested the presence of neurofilament, microtubule, ubiquitin, and paired helical filament-related epitopes in Lewy bodies. Yet the biochemical composition of the Lewy body remains incompletely elucidated. The ultrastructural and immunocytochemical similarities and differences between the Lewy body and the neurofibrillary tangle of Alzheimer's disease raise questions as to their relation to each other and possible shared mechanisms of formation. In this study the authors examine whether ultrastructural immunocytochemical analysis of Lewy bodies confirms the light microscopic data, whether the structures and epitopes of Lewy bodies share with Alzheimer's disease neurofibrillary tangles the property of insolubility in sodium dodecyl sulfate, and speculate about the subunit composition of Lewy body filaments.


Immunocytochemical characteristics of small cell lung carcinoma associated with the Lambert-Eaton myasthenic syndrome. The Lambert-Eaton myasthenic syndrome (LEMS) is characterized by the presence of IgG antibodies to motor nerve terminals, and associates with small cell lung carcinoma in more than 60% of cases. We have carried out a comparative immunocytochemical study on small cell lung carcinoma (SCLC) in five LEMS cases and six non-LEMS cases, using antibodies to tumor markers, MHC Class I and II, macrophages and lymphocytes. The authors found a reduced expression of the 200Kd neurofilament antigen and of MHC Class I antigens in the LEMS cases as well as a greater infiltration of activated macrophages. It is suggested that these findings are consistent with the view that SCLC antigenic determinants trigger the autoantibody response in SCLC-LEMS.


Nucleolar antigens and autoantibodies in hepatocellular carcinoma and other malignancies. Patients with hepatocellular carcinoma (HCC), gastrointestinal, lung, and ovarian cancers were shown to have autoantibodies to nuclear and nucleolar antigens as detected by immunofluorescence on cell substrates. The frequency of antinuclear antibodies (ANAs) was significantly higher (P less than 0.001) in patients with HCC (57/184 = 31%) than in patients with chronic hepatitis or liver cirrhosis (25/187 = 13%). Although a range of fluorescence patterns was observed, a higher percentage of nucleolar fluorescence was detected in HCC, and three of these nucleolar antigens were identified. They were NOR-90, nucleolus organizer region doublet polypeptides of 93 and 89 kDa involved in RNA polymerase I transcription; fibrillarin, a 34 kDa protein of the nucleolar U3 ribonucleoprotein particle which is engaged in preribosomal RNA processing; and nucleophosmin/protein B23, a 37 kDa polypeptide which is associated with ribosome maturation and cellular proliferation. All these antigens are nucleolar components that are engaged in some aspect of ribosome biosynthesis. Since autoantibodies to these nucleolar antigens have also been found in systemic autoimmune diseases, they do not represent autoimmune reactions unique to cancer but might reflect reaction pathways related to immune responses that are antigen-driven. The ANA response in HCC appears to be dynamic reactions to this antigen-drive since some patients with chronic liver disease showed seroconversion to ANA positivity, marked increase in titer and/or change in antibody specificity preceding or coincident with clinical detection of HCC. These changes in ANA showed a close temporal relationship with transformation from long-established chronic liver disease to HCC.


Expression of Epstein-Barr virus genes and of lymphocyte activation molecules in undifferentiated nasopharyngeal carcinomas. Previous studies investigating the role of Epstein-Barr virus (EBV) in undifferentiated nasopharyngeal carcinoma (NPC) have been performed on extracts from biopsies. The authors analyzed expression and localization of viral gene products in 18 undifferentiated NPCs at the cellular level using immunohistology and in situ hybridization. All cases were EBV-positive. The small nuclear EBV-encoded RNAs, EBERs, were regularly expressed whereas the latent membrane protein, LMP1, and EBV was detectable only in four cases (22%) and the nuclear antigen 2 was not detectable. The BZLF-1 protein of EBV which disrupts viral latency, was not detectable, confirming that the virus is latent in the tumor cells. Although the expression of the CD23 antigen in transplantable NPCs has been reported, our study demonstrates that expression of this antigen in human undifferentiated NPCs is rare. In contrast, almost all cases expressed the CDw70 antigen. Since in normal tissues this antigen is present only in activated lymphoid blasts, this finding may be relevant for the differential diagnosis of undifferentiated NPCs.


Clonal Epstein-Barr virus genome in T-cell-rich lymphomas of B or probable B lineage. Seventeen nodal lymphomas (originally diagnosed as T-cell lymphomas based on histological features and immunohistochemical staining results) were studied for the presence of Epstein-Barr virus (EBV) genome, and the results correlated with immunoglobulin and T-cell receptor gene rearrangement analyses performed on the same tissue samples. All four EBV positive cases had clonal rearrangement of the joining region of the immunoglobulin heavy chain (IgJH) gene without clonal T-cell receptor beta-chain (TCR beta) gene rearrangement. Of these, two cases also showed clonally rearranged light chain gene, and they were reclassified as T-cell rich B-cell lymphomas (TRBL). The other two cases lacked clonal kappa or lambda light chain rearrangement and they were reclassified as T-cell rich lymphomas of probable B lineage, based on their isolated IgJH clonal rearrangement. These B-cell lymphomas may be easily misdiagnosed as T-cell lymphomas owing to the presence of an abundant reactive T-cell infiltrate masking the tumor population. The florid T-cell reaction may represent an unusual host response towards a clonal proliferation of EBV bearing B cells.


Virus-associated RNAs (VA-I and VA-II). An efficient target for the detection of adenovirus infections by in situ hybridization. The identification of adenovirus in tissue can be difficult. In situ hybridization for adenovirus nucleic acids may aid in the demonstration of adenovirus infections. To develop a probe against adenovirus, a 978 bp fragment of DNA containing the VA-I, VA-II, and a portion of the L-1 regions of the adenovirus type 2 genome was cloned into the SK+ vector. These regions were selected because they are generally conserved among adenoviruses and are abundantly transcribed during the lytic cycle. Sense and antisense tritium or Digoxigenin-labeled riboprobes were generated using in vitro transcription and applied to formalin-fixed paraffin-embedded sections of HeLa cells infected with adenovirus type 2. Extensive in situ hybridization of the antisense riboprobe to HeLa cells with cytopathic changes was found. The number of cells to which the probe hybridized decreased proportionately with dilution of infected with noninfected cells. The control sense riboprobe showed only scattered breakthrough hybridization and in these cells hybridization was mainly located in the nucleus. Northern blot analysis of RNA from infected HeLa cells confirmed the in situ hybridization results. No hybridization was detected when cultured cells infected with herpes simplex virus, Epstein-Barr virus, cytomegalovirus, or human immunodeficiency virus were examined. Specific hybridization was detected in tissues obtained at autopsy from four patients with culture proven adenovirus infection. These observations suggest that this probe is useful in the diagnosis of adenovirus in formalin-fixed paraffin-embedded material.


Membranous cytoplasmic bodies in the lateral wall of the cat cochlea. Membranous cytoplasmic bodies (MCBs) were observed both in the intermediate cells of the stria vascularis and in the fibrocytes of the spiral ligament in normal cats. The MCBs in the intermediate cells were round or ovoid, ranged from 1 to 5 microns in diameter, and consisted of 2 to 40 or more layers disposed concentrically about a core. The core was composed of not only cytoplasm but also of mitochondria in various stages of disintegration, empty vacuoles, and many kinds of dense bodies similar to lysosomes. The MCBs in the fibrocytes were similar to those in the intermediate cells, but they were a little smaller in diameter and consisted of fewer than 10 layers. Both in the intermediate cells and in the fibrocytes, MCBs seem to have a phagocytic function.


Synovial sarcomas of the head and neck.

Synovial sarcomas of the head and neck. Springing from mesenchyme rather than mature synovial tissues, synovial sarcomas are high-grade neoplasms that express epithelial as well as supporting tissue features. Accordingly, their histologic phenotypes can be epithelial, stromal, or mixed. Between 3% and 10% of all synovial sarcomas originate in the head and neck, particularly from parapharyngeal sites. These is no appreciable difference in biologic activity between synovial sarcomas of the head and neck and those arising from other anatomic sites. Five-year survivals are misleadingly optimistic and do not adequately reflect the natural history of the sarcoma. Synovial sarcoma is known to metastasize late, and few patients survive that event.


Developmental changes in lung epithelial ion transport and liquid movement. During fetal life, the lungs are filled with liquid that flows from the pulmonary circulation across the epithelium in response to the osmotic force generated by Cl- secretion of airway and distal lung epithelial cells. As birth approaches, net Cl- secretion across the respiratory tract epithelium decreases, and this is associated with a reduction in the flow of liquid into the lung lumen. The cause for this change is unknown, but several recent studies indicate that it may be related to alterations in the hormonal milieu to which the lung epithelium is exposed late in gestation. The switch from placental to pulmonary gas exchange at birth requires rapid removal of liquid from the lung lumen. During labor and the immediate postnatal period, the pulmonary epithelium changes from a predominantly Cl-(-)secreting membrane to a predominantly Na(+)-absorbing membrane, with resultant reversal of the direction of flow of lung liquid. There is considerable evidence that this change reflects an active metabolic process involving increased Na(+)-K(+)-ATPase activity in lung epithelial cells, which drives liquid from the lung lumen into the interstitium, with subsequent absorption into the pulmonary circulation. This Na(+)-K(+)-ATPase-dependent process persists in the bronchopulmonary epithelium of the mature lung and probably has an important role in clearance of alveolar edema associated with heart failure or lung injury.


NMDA-receptor-independent long-term potentiation. Although NMDA-R-dep LTP in the hippocampus has received much attention, it is clear that many types of LTP do not involve NMDA receptors. While early studies of NMDA-R-indep LTP were done in invertebrates, an NMDA-R-indep LTP is also seen in at least three excitatory pathways of the hippocampus. There would appear to be quite diverse mechanisms of induction of NMDA-R-indep LTP, although in most cases there is evidence, or at least a suggestion, that Ca2+ is involved. At the hippocampal CA3 MF synapse, activation of voltage-gated Ca2+ channels has been proposed as a trigger for LTP induction, and this may also be the case for certain types of LTP at the SC synapse in CA1 (25, 40). The modulation of both MF LTP and Ca2+ channels by beta-adrenoreceptor and muscarinic agonists suggests that specifically the L-type channel is critical for MF LTP induction. L-type Ca2+ channels may also be involved in NMDA-R-indep LTP at SC synapses (6, 40). Clearly more work is needed to test these possibilities. In addition, it will be interesting to discover whether voltage-gated Ca2+ channels play a role in LTP in other areas of the brain such as the cerebral cortex and amygdala (24).


Adaptations to high hydrostatic pressure. The importance of adaptation to high pressure has long been implicit in the findings of studies in which 1 atm-adapted species were subjected to elevated pressures. Recent comparative studies have shown that pressure sensitivities of enzymes, structural proteins, and membrane-based systems differ markedly between shallow- and deep-living species. These studies allow operational definition of what constitutes high pressures for different biological structures and processes. These are the habitat (adaptation) pressures at which a given type of system first exhibits reduced perturbation by pressure. These threshold pressures vary among physiological systems, but are similar for a given system among different species. Dehydrogenase enzymes and adenylyl cyclases exhibit threshold perturbation pressures of only 50-100 atm; the Na(+)-K(+)-ATPase of teleost gills appears to have a pressure perturbation threshold near 200 atm, and a similar threshold was found for actin self-assembly. Even this limited sample of physiological processes indicates that the terms deep and high pressure begin to apply at depths of only 500 m or less--and processes yet to be examined in comparative analysis may yield even lower pressure thresholds. The differences in sensitivity to pressure of homologous systems in shallow- and deep-living organisms have implications at several levels of biological organization. The vertical distribution patterns of species in aquatic habitats may be established, in part, by interspecific differences in resistance to pressure. High pressures may restrict the depths to which shallow-living species can penetrate, and the obligately barophilic systems found in deep-living organisms may limit their upper distribution limits. The similarities noted among the adaptations of deep-sea species with different shallow-water ancestors reflect a high degree of convergent evolution in pressure adaptation. It will be interesting to learn if the similarities in pressure-resistance of function among diverse deep-sea species are the result of similar or identical changes at the molecular level, e.g. in protein sequence. Acclimation to pressure may be of widespread occurrence among species that undergo large changes in depth, e.g. during ontogeny. Pressure acclimation may require pressure-regulation of gene expression. Lastly, comparisons of species from the cold deep sea with those from hydrothermal vents have shown that adaptations to both temperature and pressure play critical roles in determining the distribution patterns of deep-living species.


Parathyroid hormone receptors in control of proximal tubule function. Parathyroid hormone action on renal proximal tubule function involves phospholipase C/protein kinase C as well as adenylate cyclase/protein kinase A mediated regulatory pathways. Tissue culture experiments suggest that low concentrations of PTH affect preferentially the phospholipase C/protein kinase C pathway. In vivo, both regulatory cascades are probably involved in the regulation of proximal tubule function. It is not clear at present whether the two intracellular pathways are linked to one or two PTH receptors. A polarized distribution of PTH receptor(s) involving different second messengers appears possible in proximal tubule epithelial cells. High-affinity (Kd 10(-11)-10(-12) M) PTH receptors in the range of circulating PTH concentrations in vivo remain to be identified. Structural and functional characterization of PTH receptors as well as of the PTH-sensitive intracellular mediators and transport systems form the basis for a better understanding of PTH-dependent regulation of proximal tubule function.


Human spumavirus antibodies in sera from African patients. Serum samples collected from patients with a wide variety of diseases from African and other countries were tested for antibodies to the human spumaretrovirus (HSRV). A spumaviral env-specific ELISA was employed as screening test. Out of 3020 human sera screened, 106 were found to be positive (3.2%). While the majority of patients' sera from Europe (1581) were negative, 26 were positive (1.6%). Sera from healthy adult blood donors (609), from patients with multiple sclerosis (48), Graves' disease (45), and chronic fatigue syndrome (41) were negative or showed a very low prevalence for spumaviral env antibodies. A higher percentage of seropositives (6.3%) were found among 1338 African patients from Tanzania, Kenya, and Gabon. Out of 1180 patients from Tanzania, 708 suffered from tumors, 75 from AIDS, and 128 had gynecological problems; 51 of the Tanzanian patients were HSRV seropositive (4.3%). A particularly high percentage of 16.6% seropositives were identified among nasopharyngeal carcinoma patients (NPC) from Kenya and Tanzania consistent with results reported 10 years ago. However, 20 nasopharyngeal carcinoma patients from Malaysia were HSRV-seronegative. In selected cases, sera from seropositive individuals were reacted with proteins from HSRV-infected cells in vitro. HSRV env- and gag-specific antibodies were specifically detected by these sera in Western blots. The results indicate spumavirus infections in human patients with various diseases at a relatively low prevalence worldwide; in African patients, however, the prevalence of spumavirus infections is markedly higher.


Monoclonal antibodies for the identification of herpesvirus simiae (B virus). To differentiate between B virus and HSV isolates from monkeys and man monoclonal antibodies (mabs) were produced to herpesvirus simiae (B virus) and herpes simplex type 1 and 2 (HSV-1 and HSV-2). Mabs were tested by indirect immunofluorescence (IFAT) for reactivity against herpesviruses from Asiatic monkeys (B virus), African monkeys (SA 8 virus), and man (HSV-1, HSV-2, varicella-zoster virus, cytomegalovirus, and Epstein-Barr virus). Mabs could be divided into groups A-E displaying specific reactivity for B virus (A); reactivity with both B virus and SA 8 but not HSV (B); reactivity with B virus, SA 8 virus and HSV strains (C); specific reactivity with HSV-1 (D); and specific reactivity with HSV-2 (E). Two of the B virus specific mabs were able to differentiate between cynomolgus and rhesus strains of B virus. None of the mabs reacted with human varicella-zoster virus, cytomegalovirus, or Epstein-Barr virus. A panel of mabs for the unequivocal identification of B virus isolates from monkey or man is proposed.


Inhibition of human immunodeficiency virus (HIV) type 1 multiplication by an avian cellular factor. A factor secreted from avian cells infected non productively with a non cytopathogenic mutant of vesicular stomatitis virus (VSV ts 1026) interferes with HIV replication in CEM cells and peripheral blood monocytes (PBL). Production of infectious particles is decreased and many virions lack cores and/or spikes. In CEM cells the prmRNA is spliced into 7.5, 4, and 2 kb mRNA. Residual virus contains less env encoded proteins and p 18; p 25 appears as several bands. The processing of tat, rev. and nef proteins differs in treated cells and in controls.


Pathogenesis of equine herpesvirus-1 in specific pathogen-free foals: primary and secondary infections and reactivation. Six specific pathogen-free foals shown to be free of equine herpesvirus-1 and 4 (EHV-1 and -4) and lacking in maternally-derived antibodies were used to investigate the pathogenesis of EHV-1 in horses. Following primary intranasal inoculation with EHV-1 all foals showed signs of a mild, self-limiting upper respiratory tract infection. A leucopenia was observed, comprising both a lymphopenia and neutropenia. Virus was isolated from nasal mucus and buffy coat cells over several days during the clinical episode and after the animals became clinically normal. Notwithstanding the mildness of the clinical disease, virus was not eliminated completely and intravenous administration of dexamethasone resulted in reactivation of latent EHV-1 in animals which had received only a single dose of the virus. In a second infection given to four foals, 61 days after the primary inoculation, no clinical signs were observed, haematological changes were minimal and viraemia was absent.


Comparison of herpesviruses isolated from reindeer, goats, and cattle by restriction endonuclease analysis. A genomic comparison of bovine herpesvirus 1 (BHV-1), caprine herpesvirus (CHV-2) and reindeer herpesvirus (RHV), was performed using 5 restriction endonucleases. Cross neutralization of these three herpesviruses showed that BHV-1 and CHV-2 had a relatively low degree of cross reaction with heterologous viruses. RHV showed a higher degree of such cross reactivity. The restriction endonuclease analyses showed that the migration patterns of the DNA segments were different for the three groups of herpesviruses. The enteric caprine strain could be differentiated from genital strains using BstE II and Hpa I. The genome size of reindeer herpesvirus was estimated to be approximately 86.8 x 10(6) Da (131.8 kbp), and indications of isomerization of this genome were found. It is concluded that reindeer herpesvirus is a distinct species within the family Herpesviridae.