Severe cytomegalovirus pneumonitis in HIV infected patients with higher than average CD4 counts. Cytomegalovirus may replicate within the lungs both of recipients of transplants and of patients infected with the human immunodeficiency virus (HIV). A hypothesis formulated by this group was that a host damaging immune response might be provoked by cytomegalovirus infection and cause a severe pneumonitis in recipients of allogeneic transplants, whereas the progressive impairment of cellular immunity in patients with HIV disease would preclude a damaging immune response in the lungs, and thus protect these patients from severe cytomegalovirus pneumonitis. This study set out to discover whether severe cytomegalovirus pneumonitis arises in HIV infected patients. Data were prospectively collected on severity of pneumonitis and infectious agents identified in consecutive respiratory episodes in HIV infected patients undergoing diagnostic bronchoalveolar lavage during 20 months. Eighty five episodes of pneumonitis occurred in 68 patients. Cytomegalovirus was identified as the only infectious agent in nine episodes (nine patients). Seven of the episodes were mild; all these patients had CD4 counts below 0.1 x 10(9)/1. The remaining two episodes were severe and ventilatory support was required. In both cases the CD4 counts were above 0.2 x 10(9)/1 and HIV infection appeared to have been acquired shortly before presentation. Although rare, severe cytomegalovirus pneumonitis may occur in HIV infected patients. Both patients with severe pneumonitis in this series had relatively well preserved immune function. These findings support the hypothesis that severe cytomegalovirus pneumonitis is an immunopathological condition.


Changes of autonomic receptors following castration and estrogen administration in the male rabbit urethral smooth muscle. Effects of castration and estrogen administration on the distribution of autonomic receptors in the male rabbit urethral smooth muscle were examined. Thirty-eight mature Japanese white rabbits were used. Eight rabbits were used as untreated control. The other 30 rabbits were castrated and 10 of them were administered intramuscularly with 5 mg/kg estrogen 3 times a week for 4 weeks from 12 weeks after castration. Autonomic receptor densities in the urethral smooth muscle were measured by radioligand binding techniques. alpha 1-Adrenergic and muscarinic cholinergic receptor densities decreased significantly after castration and were not affected by estrogen administration. alpha 2-Adrenergic receptor density increased slightly after castration and markedly after estrogen administration. beta-Adrenergic receptors were not affected either by castration or estrogen administration. No significant changes were observed in any receptor affinities. These results demonstrate the effects of castration and estrogen administration on the autonomic receptors in the male rabbit urethral smooth muscle.


DT diaphorase [NAD(P)H: (quinone acceptor) oxidoreductase] facilitates redox cycling of menadione in channel catfish (Ictalurus punctatus) cytosol. Characteristics of DT diaphorase (NAD(P)H: (quinone acceptor) oxidoreductase, DTD) activity in Ictalurus punctatus and the effect of DTD activity on menadione (MND)-mediated reduction of acetylated cytochrome c (AcC) were examined. DTD activity in cytosols of four organs followed a distinct gradient in the order stomach greater than gill greater than liver greater than posterior kidney. A similar gradient was observed in organ-specific rates of in vitro AcC reduction in the presence of either NADH or NADPH as reducing equivalent. A greater proportion of the AcC reduction rate was sensitive to inhibition by dicoumarol (DC) in organs with relatively high DTD specific activity (e.g., stomach) than in organs with low DTD activity (e.g., kidney). No such trend was observed in the superoxide dismutase (SOD)-sensitive proportion of AcC reduction rates. DTD was observed to contribute to MND-mediated superoxide production to a greater extent in organs with high DTD activity than in organs with low DTD activity. DC-sensitive (i.e., DTD-mediated) AcC reduction was observed to increase with organ-specific DTD activity, and the majority of the AcC reduction rate was inhibitable by SOD. These findings demonstrate a direct contribution by DTD activity to MND-mediated superoxide production in this in vitro system. The role of I. punctatus DTD as a possible deleterious agent in quinone metabolism and implications regarding the traditional conception of DTD as a detoxifying enzyme are discussed.


Application of the EPR spin-trapping technique to the detection of radicals produced in vivo during inhalation exposure of rats to ozone. Ozone is known to induce lipid peroxidation of lung tissue, although no direct evidence of free radical formation has been reported. We have used the electron paramagnetic resonance (EPR) spin-trapping technique to search for free radicals produced in vivo by ozone exposure. The spin trap alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (4-POBN) was administered ip to male Sprague-Dawley rats. The rats were then exposed for 2 hr to either 0, 0.5, 1.0, 1.5, or 2.0 ppm ozone with 8% CO2 to increase their respiratory rate. A six-line 4-POBN/radical spin adduct signal (aN = 15.02 G and a beta H = 3.27 G) was detected by EPR spectroscopy in lipid extracts from lungs of rats treated with 4-POBN and then exposed to ozone. Only a weak signal was observed in the corresponding solution from rats exposed to 0 ppm ozone (air with CO2 only). The concentration of the radical adduct increased as a function of ozone concentration. After administration of 4-POBN, rats were exposed for either 0.5, 1.0, 2.0, or 4.0 hr to either 0 or 2.0 ppm ozone (with CO2). The radical adduct concentration of the ozone-exposed groups at exposure times of 2.0 and 4.0 hr was significantly different from that of the corresponding air control groups. A correlation was observed between the radical adduct concentration and the lung weight/body weight ratio. These results demonstrate that ozone induces the production of free radicals in rat lungs during inhalation exposure and that radical production may be involved in the induction of pulmonary toxicity by ozone. This is the first direct evidence for ozone-induced free radical production in vivo.


Polychlorinated and polybrominated biphenyl congeners and retinoid levels in rat tissues: structure-activity relationships. The purpose of this study was to examine the structural requirements of polychlorinated and polybrominated biphenyls (PCBs and PBBs) for altering tissue levels of retinoids. Seven congeneric PCBs and PBBs were studied: 3,3',4,4'-tetrachlorobiphenyl (TCB), 2',3,3',4,5- and 3,3',4,4',5-pentachlorobiphenyls (-PeCBs), 3,3',4,4'- and 3,3',5,5'-tetrabromobiphenyls (-TBBs), 2,2',3,3',5,5'-hexachlorobiphenyl (-HCB), and 3,3',4,4',5,5'-hexabromobiphenyl (-HBB). Male Sprague-Dawley rats were fed a vitamin A-adequate diet (1.3 mg/kg) for 30 days before being given a single IP injection of one of seven polyhalogenated biphenyls (150 mumol/kg) in corn oil (10 ml/kg) or vehicle alone. Rats were killed 1 week later. Except for 3,3',4,4',5,5'-HBB, all PCBs and PBBs studied significantly decreased serum retinol levels and, except for 3,3',4,4',5,5'-HBB and 2,2',3,3',5,5'-HCB, all PCBs and PBBs also lowered the serum retinol-binding-protein (RBP) content. The activity of hepatic retinyl ester hydrolase (REH) was reduced by the treatment of 3,3',4,4',5-PeCB, 3,3',4,4'-TBB, and 3,3',4,4',5,5'-HBB. The levels of hepatic retinol were decreased by 2,2',3,3',5,5'-HCB, 2',3,3',4,5-PeCB, and 3,3',4,4',5-PeCB, while levels of hepatic retinyl palmitate were decreased by 2',3,3',4,5-PeCB, 3,3',4,4',5-PeCB, 3,3',4,4'-TCB, 3,3',4,4'-TBB, and 3,3',4,4',5,5'-HBB. The substantial decreases in hepatic retinyl palmitate levels could not be explained solely on the basis of hepatomegaly caused by acutely toxic PCBs and PBBs. All halogenated biphenyls which caused a decrease in hepatic retinyl palmitate also caused an increase in renal retinyl palmitate except 3,3',4,4',5-PeCB. In summary, the acutely toxic (nonortho substituted) congeners had pronounced effects on hepatic, renal, and serum retinoids whereas other biphenyls only decreased serum retinol levels. The effects of these seven compounds on REH activity were not correlated with the effects on serum retinol or RBP levels. Therefore, this study shows that the structure-activity relationships for altering hepatic retinoids differ from those for serum retinol, implying the involvement of multiple mechanisms.


Adrenal cortex, tumor, and peripheral production of deoxycorticosterone. A method is reported for the measurement of the urine excretion rates of tetrahydro-11-deoxycorticosterone (3 alpha,5 beta-THDOC), an important metabolite of 11-deoxycorticosterone (DOC). Quantification using gas chromatography/mass spectrometry (GC/MS) was achieved by comparing the ion fragment response for the molecular ion (m/z 507) of the analyte (as methyloxime trimethylsilyl ether derivative) to that of a fixed amount of an isomer of THDOC added to urine as internal standard. To improve the specificity of measuring THDOC in clinical samples, an additional Sephadex LH-20 chromatography step was introduced to separate 11-deoxycortisol and some progesterone metabolites. In the luteal phase of the menstrual cycle, THDOC excretion was higher than in the follicular phase; it was also higher than in women taking oral contraceptives. The correlation of THDOC with progesterone production, independent of a constant cortisol output, supports an ovarian or peripheral conversion of progesterone to DOC. The assay proved useful (1) in monitoring for the recurrence of a mineralocorticoid-secreting tumor and (2) when adrenal production of DOC was not fully suppressed in congenital adrenal hyperplasia due to 11 beta-hydroxylase deficiency. Under the latter circumstances, the renin-angiotensin system seemed to be an important regulator of DOC production.


Influence of ST 789 on human neutrophil function in vitro. We report the results of our in vitro experiments on the effects of an L-Arg terminal synthetic hypoxanthine derivative (ST 789) on human neutrophil function. To verify the hypothesis that the reported immunomodulatory effects of ST 789 in vivo are accounted for, at least in part, by effects on phagocytic cells, we experimented in parallel also with methisoprinol, a well known stimulator of neutrophil chemotaxis, structurally related to ST 789. Our results demonstrate that ST 789 is able to improve the true chemotactic response of human neutrophils without interfering with other phagocytic functions, following a pattern largely shared by methisoprinol. Accordingly, ST 789 may be able, when used in vivo, to prime neutrophils for timely and efficient migration to inflammatory sites, and it could be considered for therapeutic use in patients with impaired inflammatory response or severe infections.


From ligand binding to gene expression: new insights into the regulation of G-protein-coupled receptors. Transmembrane signaling systems relay information from the exterior to the interior of a cell, through a series of complex protein-protein interactions and second messenger cascades. One such system consists of the G-protein-coupled receptors, which interact with G proteins upon ligand binding, and in turn activate an effector molecule. The receptor is the first component in this signaling cascade and is subject to considerable regulation. Recent studies have shown that these regulatory events can occur at the levels of receptor protein modification and receptor gene expression. Interestingly, some of these processes appear to be mediated by the same second messenger systems that these receptors activate, which leads to various forms of positive and negative feedback regulation.


Proofreading, NTPases and translation: constraints on accurate biochemistry. Two accurate individual reactions can work together as a more accurate overall mechanism. This is straightforward for a transient reaction, but the same accuracy in the steady state (termed proofreading) requires many preconditions. The preconditions for proofreading can be summarized as four statements, which allow experimental identification and subsequent confirmation of proofreading mechanisms.


The effect of cold storage of rat thoracic aortic rings in organ preservation solutions--a study of receptor-linked vascular prostacyclin synthesis. An important aspect of organ preservation is the maintenance of intrinsic dilator and antithrombotic mechanisms of blood vessels. Blood vessels synthesize prostacyclin (PGI2), a potent vasodilator and inhibitor of platelet adhesion and aggregation. PGI2 synthesis is controlled by complex mechanisms including adrenoceptor-linked calcium influx and protein kinase C. Since organ preservation solutions may influence these mechanisms, we investigated the effect on in vitro PGI2 synthesis of cold storage of rat aortic rings in lactobionate-raffinose solution (LRS) and hypertonic citrate kidney preservation solution (KPS) on in vitro PGI2 synthesis. Acute incubation of aortic tissue in both preservation solutions at 37 degrees C (compared with minimal essential medium) completely inhibited PGI2 synthesis when stimulated with noradrenaline (NA), phorbol ester (a protein kinase C activator), NaF (a G protein activator), or A23187. Following storage of aortic rings at 4 degrees C (for up to 72 hr) in LRS and KPS, subsequent washing and incubation in MEM, PGI2 synthesis was initially markedly enhanced in response to NA when compared with tissues stored in MEM. These enhanced responses disappeared, and PGI2 synthesis returned to normal following 1 hr incubation of tissues in MEM at 37 degrees C. These data demonstrate that cold storage in preservation fluids exerts minimal deleterious effects, not only on PGI2 synthesis, but possibly on other key processes (calcium homeostasis, protein kinase C activity) in blood vessels.


Mitochondrial diseases and myopathies: a series of muscle biopsy specimens with ultrastructural changes in the mitochondria. From 1986 to 1991, 472 muscle biopsy specimens from patients from different hospitals in Norway were examined. Of these, 364 were embedded for electron microscopy, and 194 were examined with electron microscopy. Ultrastructural alterations in the mitochondria were detected in 49 of these specimens. Characteristic electron microscopic findings included subsarcolemmal accumulation of abnormal mitochondria of various shapes and sizes, often containing electron-dense granules and sometimes lipid vacuoles in the mitochondria and diffusely electron-lucent matrix space. Paracrystalline inclusion bodies were seldom seen in specimens from young patients, but in some cases mitochondrial electron-dense granules at the cristae were found. These amorphous densities are consistent with lipoproteins, suggesting that they may represent an early stage of paracrystalline inclusions. Biochemical and genetic exploration of the patients with biopsy specimens suggesting mitochondrial disease indicated maternally genetic inheritance and an enzyme defect in the respiratory chain in 21 patients in two families. Three patients had MELAS syndrome, 7 Marinesco-Sjögren syndrome, and 2 Kearns-Sayre syndrome. Five family members had ptosis, cardiomyopathy, mild myopathy, and increased lactate in cerebrospinal fluid and serum. In addition to the diseases mentioned above, changes in the mitochondria were detected in other conditions such as Rett's syndrome (n = 1), ornithine transcarbamylase deficiency (n = 2), and hypothyroidism (n = 2) as well as in 3 patients with clinical and laboratory results indicative of inflammatory myopathy and 3 patients with clinical and laboratory findings consistent with peripheral neuropathy. It is concluded that, although ultrastructural changes in the mitochondria may represent unspecific findings, electron microscopic examination of muscle biopsy specimens is a useful screening method to select specimens for further biochemical analysis and to obtain an early and more precise diagnosis of the disease.


Lennert's lymphoma with giant multivesicular lysosomal bodies optically visible. A case of a 43-year-old male patient with lymphoepithelioid lymphoma is discussed. During the 10-year follow-up of this patient, different biopsy specimens were obtained. Immunohistochemical studies with a complete paraffin panel determined a T-cell nature of the specimens. Electron microscopy showed a peculiar finding in the cytoplasms of the tumoral cells that correlated with optical inclusions observed in optical imprints; these imprints corresponded to lysosomic multivesicular bodies, whose significance is discussed.


Anaplastic large cell Ki-1 lymphoma of the stomach with villopodial projections: an immunocytochemical and ultrastructural study and review of the literature. The report describes a case of anaplastic large cell Ki-1 lymphoma of the stomach with a rapidly fatal course observed in a 28-year-old woman. By electron microscopy the neoplastic cells presented long projections of the cytoplasmic membrane arranged uniformly along all the cell circumference. The morphology and distribution of these projections were characteristic, and the term villopodial is proposed for them. A review of the literature revealed 18 cases of lymphoma and 7 cases of nonlymphoid neoplasms composed of cells with projections similar to those observed in the present case. It is suggested that such tumors be denominated villopodial lymphomas and villopodial tumors. The present case is the first anaplastic large cell Ki-1 lymphoma with villopodial projections reported in the literature.


Effect of alkalinization on calcium oxalate monohydrate calculi during extracorporeal shock wave lithotripsy: in vivo experiments. Previous in vitro experiments demonstrated the reduced microhardness of calcium oxalate monohydrate (COM) calculi, relative to dry values, when saturated with an alkaline solution (pH = 9.5). Nineteen patients with a COM calculus in the distal ureter which had been resistant to prior extracorporeal shock wave lithotripsy in situ, were treated when the stone was surrounded by alkaline urine. The urine of 14 patients was alkalinized orally by administration of acetazolamine and citrate solution; in 5 other patients direct percutaneous irrigation of sodium bicarbonate via a nephrostomy tube was carried out. The urinary pH just before lithotripsy was greater than or equal to 9 in 17/19 patients. 4,000 shock waves, averaging 18.1 kV generated by the Siemens Lithostar, were delivered onto the calculus. No significant increase of comminution rate was apparent at radiographic control immediately after the treatment and only in half of the cases was evacuation obtained within 3 months.


Duct-acinar-islet cell tumor of the pancreas. Two cases of pancreatic tumor consisting of duct, acinar, and islet components are reported. Both tumors measured about 1.0 cm in diameter and were without definite fibrous encapsulation. Histologic, immunocytochemical, and electron microscopic studies revealed three distinct cell populations: duct, acinar, and islet cells. Both endocrine and exocrine components were seen within the same cell nest. Islet components predominated in both cases. Nearly all the cells in the islet component were positive for insulin. Few cells positive for glucagon, somatostatin, or pancreatic polypeptide were present within the tumor cell nests. Duct cells were the least conspicuous cellular element of the tumor; they were positive for mucin and immunoreactive for cytokeratin and carcinoembryonic antigens (CEA). The acinar component was the minor element of the tumor in both cases. Electron microscopic study also confirmed three different cell populations in the tumor: duct cells arranged in a ductal structure with intercellular attachments and microvilli, islet cells containing beta granules, and acinar cells with zymogen granules. The tumors presented herein indicate that both their endocrine and exocrine components might have been derived from a common precursor. The implication and significance of the differentiation of different cells within the same tumor is discussed in relation to the concept of an amine precursor uptake and decarboxylation (APUD) system.


Presence of human papillomavirus 6/11 DNA in condyloma acuminatum of the urinary bladder. A Japanese woman with condyloma acuminatum of the urinary bladder is presented. The condyloma acuminatum lesion was resected endoscopically and human papillomavirus 6/11 DNA was detected. After treatment, there has been no recurrence of the disease.


Sclerosing stromal tumor of the ovary: an ultrastructural and immunohistochemical analysis with histogenetic considerations. Sclerosing stromal tumors are rare, benign ovarian neoplasms of unknown etiology and histogenesis. Three sclerosing stromal tumors were evaluated by immunohistochemistry and electron microscopy and were compared to two thecomas and nonneoplastic ovarian mesenchymal tissue. The sclerosing stromal tumors and thecomas were positive for muscle-specific actin; immunoreactivity was intense in the cellular areas of the sclerosing stromal tumors and focal in the thecomas. This antigen was expressed in nonneoplastic stroma predominantly in a perifollicular (theca externa) distribution. Two sclerosing stromal tumors and both thecomas were vimentin positive. Desmin was present in nonvascular cells in one of each tumor type. Expression of vimentin diffusely and of desmin focally was present in nonneoplastic cortical stroma and surrounding follicles. All specimens were nonreactive for cytokeratin. Electron microscopy supported differentiation toward smooth muscle in the sclerosing stromal tumors but not in the thecomas. Such differentiation included aggregates of cytoplasmic filaments with interspersed dense bodies, pinocytotic vesicles, and basal lamina. Delicate, long processes interconnected cells, often with primitive junctions, in the hypocellular foci. Cytoplasmic lipid, which was present in the thecomas, was not well developed in the sclerosing stromal tumors. It is proposed that a population of muscle-specific actin-positive elements exists in the theca externa--the perifollicular myoid stromal cell--and that sclerosing stromal tumors may originate from them. Sclerosing stromal tumors and thecomas share many antigenic determinants and morphologic features and thus are probably closely related entities.


Alveolar cell carcinoma presenting as malignant pericardial effusion: diagnosis by electron microscopy. The cytologic, immunocytochemical, and ultrastructural findings in a 68-year-old man who presented with a malignant pericardial effusion are reported. Radiologic studies failed to identify a primary neoplasm over the next 6 months. Ultrastructural examination of a repeat pericardiocentesis fluid specimen revealed cells with intranuclear tubular inclusions and cytoplasmic lamellar bodies typical of alveolar cell carcinoma. Review of the chest radiographs showed nonresolving patchy infiltrates in the upper lobe of the left lung clinically thought to represent pneumonia; in retrospect, however, these were consistent with the pneumonic form of alveolar cell carcinoma.


Reovirus type 2 in domestic cats: isolation and experimental transmission. Five reovirus isolates were recovered in MA104 cell cultures from the faeces of three cats with nictitating membrane protrusion and diarrhoea, one cat with diarrhoea only and from one healthy cat. Four of these isolates were characterised as reovirus type 2 and one as reovirus type 3 by haemagglutination-inhibition and serum neutralization tests. Reovirus type 2 has not been reported previously in cats. Mild clinical signs of diarrhoea were noted in kittens infected experimentally with one of the feline reovirus type 2 isolates.


Characterization of the glycoprotein D gene products of equine herpesvirus 1 using a prokaryotic cell expression vector. The gene encoding equine herpesvirus 1 (equine abortion virus; EHV-1) glycoprotein D was engineered into the prokaryotic vector pEX, and expressed as a beta-galactosidase fusion product, which was recognized by pooled equine sera and anti-EHV-1 rabbit sera. Antibodies raised against the EHV-1 gD fusion product identified strong bands in infected cells at 66 and 68 K and at 138 K in purified virus, thus characterizing the several forms of this major envelope glycoprotein which is an important candidate for inclusion in subunit vaccines.


Molecular mechanisms of visna virus Tat: identification of the targets for transcriptional activation and evidence for a post-transcriptional effect. Visna virus is a pathogenic lentivirus of sheep that is distantly related to the primate lentiviruses, including the human immunodeficiency virus type 1 (HIV-1). Replication of HIV-1 in cell culture requires the expression of a virus-encoded protein, Tat, which is a potent trans-activator of viral gene expression. Visna virus encodes an analogous Tat protein that greatly increases gene expression directed by the visna viral LTR. This report uses a stable vero cell line that constitutively expresses visna virus Tat to investigate the molecular mechanism of action of Tat on viral gene expression. Transient expression assays, using the visna virus LTR to drive transcription of the bacterial gene for chloramphenicol acetyltransferase (CAT), demonstrate that Tat trans-activates gene expression by increasing steady-state mRNA levels. The increase in steady-state mRNA levels is sufficient to account for the increase in protein observed and is due, in part, to an increase in the rate of transcription initiation. Tat mediates the accumulation of mRNA through AP-4 and AP-1 binding sites located in the U3 region of the LTR. Deletion of the upstream AP-1 and AP-4 binding sites results in a residual low level of trans-activation by Tat. Further experiments, using LTRs with R-U5 sequences deleted to +10, demonstrate AP-1 and AP-4 mediated responses to TAT at the RNA level, but no increase was observed in CAT protein.


Rapid in vivo induction of HIV-specific CD8+ cytotoxic T lymphocytes by a 15-amino acid unmodified free peptide from the immunodominant V3-loop of GP120. Efforts to generate a vaccine to prevent infection by human immunodeficiency virus (HIV) have focused on inducing neutralizing antibodies. However, cytotoxic T-lymphocyte (CTL) responses are a major immune defense mechanism required for recovery from many different virus infections. Since CTL epitopes can be defined by short synthetic peptides, we searched for HIV peptides that elicit a viral-specific CTL response in mice. We have developed a new method for screening CTL-inducing peptides involving a single injection into the footpad of mice to prime CTLs in the draining popliteal lymph node of mice within 10 days. Our results demonstrate that a 15-amino acid peptide (aa 315-329) derived from the V3 loop of HIV gp120 caused a rapid induction of peptide-specific and gp160-specific CD8-positive CTLs. Lysis of targets is specific since cells preincubated with unrelated peptides are resistant to lysis as are cells of a different MHC haplotype pretreated with the cognate peptide. Pretreatment of restimulated node cells with complement plus anti-CD8 but not anti-CD4 removed the lytic activity. We also successfully induced in vivo CTL activity with unmodified synthetic peptides from the influenza and Sendai virus nucleoproteins, indicating general applicability of our method for rapid screening of CTL epitopes. Because HIV replication has been reported by several labs to occur mainly in lymph nodes of infected patients, the rapid induction of HIV-specific CTLs in proximal lymph nodes by unmodified peptides emphasizes the physiological significance of our findings toward vaccine and therapeutic approaches.


An isoform variant of the cytomegalovirus immediate-early auto repressor functions as a transcriptional activator. The major immediate-early promoter (MIEP) of human cytomegalovirus directs the expression of several differentially spliced and polyadenylated mRNAs. These mRNAs encode nuclear phosphorproteins (IE55, IE72, and IE86), which consist of common and unique amino acid sequences. To date, very little is known of the functional role of the 55-kDa (IE55) protein. Here we present evidence that the IE55 protein is a positive activator of the MIEP. In human fibroblast cells IE55 protein activated the MIEP between 10- and 30-fold. Fusion of IE55 to the GAL4 DNA binding domain resulted in a chimeric protein capable of trans-activating a reporter with GAL4 recognition sequences. These results strongly suggest that IE55 is a bona fide transcriptional activator protein. In addition, the IE55 protein was found not to act synergistically with the IE72 activator protein. The IE55 protein shares the same amino acid sequence as IE86 except for a 154-amino-acid deletion at the C-terminal end of the protein. These proteins were functionally antagonistic; IE55 relieved repression by IE86 and, conversely, IE86 negated IE55 activation. Mutagenesis of the MIEP revealed that the target sequence for activation by IE55 is different from the IE86 autorepressive response element. These experiments suggest that the mechanism of action of the IE55 and IE86 isoforms is distinct. Moreover, from these results it is apparent that the interplay of these factors might be critical in determining the level of HCMV replication in the host.


CPF-DD is an inhibitor of infection by human immunodeficiency virus and other enveloped viruses in vitro. The initial step in the infection cycle of human immunodeficiency virus type 1 (HIV-1) involves binding of its surface glycoprotein gp 120 to the T lymphocyte CD4 antigen. CPF-DD is a low molecular weight inhibitor of HIV infectivity that inhibits gp 120 binding to CD4 in vitro (Finberg et al., Science 249, 287-291, 1990). We find, however, that the actions of CPF-DD are not limited to its ability to interfere with gp 120-CD4 binding; its predominant action is to remove the viral envelope from the underlying core. Subsequently the virions disintegrate. Most enveloped viruses tested were inhibited by CPF-DD, but the infectivity of noneneloped viruses was unaffected or only slightly reduced.


Open reading frames encoding a protein kinase, homolog of glycoprotein gX of pseudorabies virus, and a novel glycoprotein map within the unique short segment of equine herpesvirus type 1. DNA sequence analysis of the unique short (Us) segment of the genome of equine herpesvirus type 1 Kentucky A strain (EHV-1) by our laboratory and strains Kentucky D and AB1 by other workers identifies a total of nine open reading frames (ORF). In this report, we present the DNA sequence of three of these newly identified ORFs, designated EUS 2, EUS 3, and EUS 4. The EUS 2 ORF is 1146 nucleotides (nt) in length and encodes a potential protein of 382 amino acids. Cis-regulatory sequences upstream of the putative ATG start codon include a G/C box 112 nt upstream and two potential TATA-like elements located between 15 and 90 nt before the ATG. The EUS 2 translation product exhibits significant homology to Ser/Thr protein kinases encoded within the Us segments of other herpesviruses, such as herpes simplex virus (26% homology) and pseudorabies virus (PRV), (45% homology), and possesses sequence domains conserved in protein kinases of cellular and viral origin. The EUS 3 ORF begins 127 nt downstream from the EUS 2 stop codon and ends at a stop codon 1119 nt further downstream. A single TATA-like element maps 61 nt upstream of the ORF. This ORF encodes a potential protein of 373 amino acids and is a homolog of glycoprotein gX of PRV, as judged by overall homology of amino acid residues, cysteine displacement, and presence of potential glycosylation sites and signal sequence. Interestingly, the EUS 4 ORF encodes a potential membrane glycoprotein that does not exhibit homology to any reported protein sequence. The EUS 4 ORF encodes a 383 amino acid polypeptide with a sequence indicative of a signal sequence at its amino terminal end, glycosylation sites for N-linked oligosaccharides, and a transmembrane domain near its carboxyl terminus. Several cis-acting regulatory sequences lie upstream of this ORF. These findings support the observation that the short region of alphaherpesviruses show considerable variation in their genetic content and gene organization.


Structure and pathogenicity of individual variants within an immunodeficiency disease-inducing isolate of FeLV. We previously described the molecular cloning of a replication-defective variant of feline leukemia virus (FeLV) that induced fatal immunodeficiency in cats. Eighteen proviruses have now been molecularly cloned from cats inoculated with the original isolate (FeLV-FAIDS) or its in vivo passages. Three were replication-competent and each of these was noncytopathic for the feline T-cell line, 3201. Replication of the prototype, FeLV-61E, in cats was associated with development of T cell tumors in some cats. The remaining 15 proviruses were replication-defective, but each of six of these tested was found to be cytopathic for 3201 cells when rescued with the noncytopathic helper virus, 61E. Three defective/helper virus mixtures were inoculated into cats and all induced fatal immunodeficiency, but with varied efficiency and kinetics. Each of these virus mixtures was attenuated relative to a mixture containing 61E and the intestine-targeted, FeLV-FAIDS-61C prototype defective molecular clone. Furthermore, one replication-competent virus chimera generated using the envelope and LTR of the defective pathogenic variant was incapable of inducing viremia in cats. The observed differences in the biological activity between the defective viruses could be attributed to no more than 10 scattered amino acid changes in envelope and either one or two nucleotide changes in the LTR.


Molecular cloning, sequence analysis, in vitro expression, and immunoprecipitation of the major inner capsid protein of the IDIR strain of group B rotavirus (GBR). The sixth genomic segment of the infectious diarrhea of infant rats (IDIR) strain of group B rotavirus (GBR) was cloned from double-stranded RNA purified from infected rat feces. Sequence comparison with group A rotaviruses (GAR) and the human ADRV strain of GBR indicated that IDIR gene 6 encoded the major inner capsid protein. The nucleic acid sequences of the two GBR genes were 72.9% conserved, and 83.4% of the amino acids were identical. Sequence substitutions between IDIR and ADRV were more numerous than reported for heterologous GAR strains, indicating that the two GBR strains may have diverged from one another over a longer period of time. Despite the sequence heterogeneity exhibited by the major inner capsid proteins of ADRV and IDIR, hydrophilicity plots of the two gene products were nearly indistinguishable. The GBR hydrophilicity plots displayed little similarity with those of rotavirus groups A or C, indicating substantial differences in the structures of those major inner capsid proteins. In vitro transcription and translation of IDIR gene 6 yielded a polypeptide product consistent in size with that predicted from the deduced amino acid sequence and the virion major inner capsid protein. The IDIR 6 polypeptide was immunoprecipitated by antisera directed against IDIR as well as antisera directed against ADRV and a heterologous bovine strain of GBR. No immunoprecipitation was observed with control sera or antisera directed against GAR. These results confirmed that group-specific epitopes were displayed by the major inner capsid protein encoded by IDIR gene 6. Reactivity with heterologous GBR antisera also indicated that the IDIR gene 6 product may prove useful as a standard reagent in immunoassays for the detection of GBR.


Stepwise phosphorylation of vesicular stomatitis virus P protein by virion-associated kinases and uncoupling of second step from in vitro transcription. Transcription-competent cores of vesicular stomatitis virus (VSV) contain two tightly bound protein kinase activities capable of phosphorylating the viral P protein (Beckes and Perrault, Virology 184, 383-386, 1991). We examined here the specificity of these kinases for the P protein substrate and their activity during the in vitro transcription process. Conditions favoring the VSVK1 kinase activity resulted in phosphorylation of the P1 species predominantly whereas conditions favoring VSVK2, or transcription conditions, led to an increase in the proportion of the faster migrating P2 and P3 species. A minimum of 2 mol phosphate/mol P protein was incorporated in 1 hr under optimal transcription conditions. Pulse-chase experiments revealed that the VSVK2 activity converted phosphorylated P1 to P2/P3 species. Most or all of the sites modified by VSVK1 (serines only) mapped to the 78 amino acid-long N-terminal fragment of the P protein; additional serine acceptor sites of undetermined location were also phosphorylated under VSVK2 conditions. Pretreatment of virion cores with 5'-p-fluorosulfonylbenzoyl adenosine had little or no effect on P1 phosphorylation but inhibited P1 to P2/P3 conversion nearly completely, with no effect on subsequent transcription. Likewise, the addition of cell extracts had relatively little effect on P1 phosphorylation but strongly inhibited the appearance of P2/P3, without affecting concurrent transcription. We conclude that phosphorylation of the P protein during transcription in vitro is a two-step process carried out by two distinct kinase activities, but only the first step may be essential for viral mRNA synthesis.


TGEV corona virus ORF4 encodes a membrane protein that is incorporated into virions. The coding potential of the open reading frame ORF4 (82 amino acids) of transmissible gastroenteritis virus (TGEV) has been confirmed by expression using a baculovirus vector. Five monoclonal antibodies (MAbs) raised against the 10K recombinant product immunoprecipitated a polypeptide of a similar size in TGEV-infected cells. Immunofluorescence assays performed both on insect and mammalian cells revealed that ORF4 was a membrane-associated protein, a finding consistent with the prediction of a membrane-spanning segment in ORF4 sequence. Two epitopes were localized within the last 21 C-terminal residues of the sequence through peptide scanning and analysis of the reactivity of a truncated ORF4 recombinant protein. Since the relevant MAbs were found to induce a cell surface fluorescence, these data suggest that ORF4 may be an integral membrane protein having a Cexo-Nendo orientation. Anti-ORF4 MAbs were also used to show that ORF4 polypeptide may be detected in TGEV virion preparations, with an estimated number of 20 molecules incorporated per particle. Comparison of amino acid sequence data provided strong evidence that other coronaviruses encode a polypeptide homologous to TGEV ORF4. Our results led us to propose that ORF4 represents a novel minor structural polypeptide, tentatively designated SM (small membrane protein).


Immunocytochemical localization of capsid-related particles in subcellular fractions of poliovirus-infected cells. The structural proteins of poliovirus can assemble into a series of different configurations (capsid-related particles, CRP). Only some seem to be true capsid precursors and the role of most CRP in morphogenesis is unclear. We used electron microscopic immunocytochemistry with monoclonal antibodies recognizing different CRP [protomers, pentamers, 65S empty capsids (EC), 74S-EC, and virions] to locate CRP in subcellular fractions containing virus-induced vesicles associated with the viral replication complex. We found pentamer antigenic CRP to be associated with the replication complex. The same pentamer antigenicity was exhibited by novel, "capsid-like" structures attached to the surface of the virus-induced vesicles. Upon solubilization of the vesicular fraction, mainly 65S-EC and only negligible amounts of pentamers were found by sucrose gradient analysis and by immunoprecipitation. We show that the pentamer antigenic particles are converted into 65S-EC when their membranous support is dissolved. We propose that the vesicular membrane prevents the assembly of 65S-EC and keeps the pentamer antigenic CRP in the appropriate concentration and configuration for association with the nascent progeny RNA.


Initiation of translation of human rhinovirus RNA: mapping the internal ribosome entry site. In order to map the 3' boundary of the segments needed for translation initiation at the correct site on human rhinovirus 2, deletions were made from the 3' end of the viral 5'-untranslated region. These truncated viral segments were placed immediately upstream of a reporter gene, a derivative of the influenza virus NS cDNA, either as monocistronic constructs or as dicistronic constructs in which the upstream cistron was the Xenopus laevis cyclin B2 cDNA. In vitro transcripts of these clones were translated in the rabbit reticulocyte lysate system, with or without supplementation with crude HeLa cell initiation factors, or in a HeLa cell-free system. When the full-length viral 5'-untranslated region was present, the HeLa cell factors strongly stimulated the synthesis of the NS-related polypeptides, especially in the case of the dicistronic mRNAs. Deletions from the 3' end extending up to nt 562 had little effect on translation efficiency or the response to HeLa cell factors, but more extensive deletions resulted in the complete loss of response to these factors, an almost total inhibition of NS synthesis from dicistronic mRNAs, and a partial inhibition in the case of the monocistronic mRNAs. In the case of a deletion extending to nt 554, insertion of a 15 nucleotide residue linker failed to restore efficient translation initiation. We conclude that the essential sequences for internal initiation extend to a point located between nt 554 and 562, and that the ribosome entry site, defined as the most 5'-proximal point where the ribosome can bind in an initiation-competent manner, must lie within 6 residues on either side of nt 562, and certainly not further downstream than nt 568.


Identification and characterization of an equine herpesvirus 1 late gene encoding a potential zinc finger. In this report, we present the DNA sequence and transcriptional characterization of a gene (IR5) that maps within each of the inverted repeat (IR) segments of the equine herpesvirus type 1 (EHV-1) genome. The IR5 open reading frame (ORF) is located within both IR sequences (nucleotides 9932-10,642 of the IR). DNA sequence analyses of the IR5 gene region revealed an ORF of 236 amino acids (24,793 Da) that showed significant homology to ORF64 of varicella-zoster virus and ORF3 of EHV-4 both of which map within the inverted repeats and to the US10 ORF of herpes simplex virus type 1 (HSV-1) which maps within the unique short segment. Additional analyses of the nucleotide sequence failed to reveal any overlapping ORFs that would correspond to US11 or US12 of HSV-1. Interestingly, the IR5 ORF of EHV-1 possesses a sequence of 13 amino acids (CAYWCCLGHAFAC) that is a perfect match to the consensus zinc finger motif (C-X2-4-C-X2-15-C/H-X2-4-C/H). Putative cis-acting elements flanking the IR5 ORF include a TATA box (nucleotides 9864-9870), a CAAT box (nucleotides 9709-9714), and a polyadenylation signal (nucleotides 10,645-10,650). Northern blot and S1 nuclease analyses identified a single 0.9-kb mRNA species that first appears at 2 hr postinfection, and whose synthesis is reduced in the presence of phosphonoacetic acid, an inhibitor of EHV-1 DNA synthesis. Thus, the IR5 gene of EHV-1 exhibits characteristics representative of a late gene of the gamma-1 class. The characterization of the IR5 gene at the DNA and RNA levels will facilitate ongoing studies to identify and characterize the IR5 polypeptide.


Alterations within pp59v-rel-containing protein complexes following the stimulation of REV-T-transformed lymphoid cells with zinc. pp59v-rel exists in association with specific cellular proteins within lymphoid cells transformed by reticuloendotheliosis virus (REV-T). These include the cellular rel homolog (p75c-rel) as well as a 40-kDa avian homolog to I kappa B. The brief exposure of REV-T-transformed lymphoid cells to micromolar concentrations of ZnSO4 induces profound alterations within these protein complexes. Most of the constituents of the rel protein complexes (to include pp59v-rel, p75c-rel, and p115) translocate from the cytosol to the nucleus. This system has been used to characterize the molecular events that accompany the activation of rel protein complexes. The level of phosphorylation increases on three proteins within these complexes: pp59v-rel, p75-c-rel, and pp40. The degree of phosphorylation on pp59v-rel is such that its relative mass increases 3 to 6 kDa when resolved by SDS-polyacrylamide gel electrophoresis. pp59v-rel is phosphorylated on serine and threonine residues predominantly within a single domain of 17.5 kDa. Similarly, p75c-rel exhibits a corresponding increase in its relative mass with increased phosphorylation. The increased phosphorylation of pp40 is accompanied by its dissociation from the cytosolic rel protein complexes. These observations draw parallels with the induction of the NF-kappa B trans-activating factor.


Characterization of the hepatitis C virus E2/NS1 gene product expressed in mammalian cells. Truncated and full-length versions of the hepatitis C virus protein domain encoding a presumptive envelope glycoprotein designated E2/NS1 were stably expressed in CHO cell lines. Characterization of the processing events involved in the maturation of E2/NS1 revealed that a high-mannose form resident in the endoplasmic reticulum was the most abundant form detected intracellularly. The ionophore carboxyl cyanide m-chlorophenyl-hydrazone was used to show that the E2/NS1 glycoprotein resided in the endoplasmic reticulum. The full-length form of E2/NS1 appeared to be cell-associated and could not be detected as a secreted product. C-terminal truncated molecules could be detected in the extracellular media as fully processed glycoproteins containing terminal sialic acid additions. These truncated glycoproteins are predicted to be biologically relevant targets of the host immune response and are therefore potential subunit vaccine candidates.


Infection of the HTLV-I-harbouring T-lymphoblastoid line MT-2 by Epstein-Barr virus. Epstein-Barr virus (EBV), a ubiquitous human B-lymphotropic virus, is associated with certain lymphoproliferative diseases of T-cell lineage. To understand the mechanism by which EBV infects T cells, we have tested the susceptibility of various human T-cell lines to the virus. We report here that the HTLV-I-harbouring T-lymphoblastoid line MT-2 carries a high level of CD21/EBV receptors on their surface, adsorbs fluorescein isothiocyanate-labeled EBV, and synthesizes virus latent antigens (EBNA-1 and LMP) following EBV infection. Pretreatment of MT-2 cells with anti-CD21 monoclonal antibody OKB7 inhibited the virus binding as well as the synthesis of virus latent antigen. These data suggest that human T-cells can be infected with EBV via functionally active virus receptors.


Cloning and sequence analysis of the genes encoding the nonstructural proteins of Langat virus and comparative analysis with other flaviviruses. Langat virus, a member of the family Flaviviridae is antigenically very similar to highly pathogenic tick-borne encephalitis viruses. We cloned and sequenced the complete nonstructural gene-coding region of Langat virus (strain TP21) and compared the deduced amino acid sequences of each nonstructural protein to those of other flaviviruses. By alignment with the reported amino acid sequences of the nonstructural proteins of several flaviviruses, we were able to predict proteolytic cleavage sites and identify sequence motifs, which are highly conserved among flaviviruses. Sequence similarity calculations revealed that the NS3 and NS5 proteins are the most highly conserved of the flavivirus nonstructural proteins. The NS3 and NS5 proteins of Langat virus contained specific peptide sequences that have been demonstrated to be associated with helicase or polymerase activities, respectively. The NS1 protein of Langat virus displayed complete homology of potential N-linked glycosylation sites and cysteine residues with the NS1 proteins of other tick-borne flaviviruses, suggesting a highly conserved NS1 protein structure. The data presented in this report serve to complete the entire sequence of the Langat virus-coding region and provide the basis for comparison of this naturally attenuated virus to the other highly virulent tick-borne flaviviruses.


Characterization of the avian adenovirus penton base. A portion of the fowl adenovirus 10 (FAV-10) penton base gene was located within a Sau3A genomic DNA fragment by the combination of a plasmid expression library and colony immunoassays with rabbit anti-FAV-10-sera. The coding portion of the sequence contained in the expression vector was mapped to a 11.6-kb Bg/II fragment and more precisely mapped to the right-hand end of the 11.6-kb fragment at map units 37.7 to 41.3. Nucleotide sequence analysis of the region revealed an open reading frame of 1575 bp with translation of the sequence producing a polypeptide of 525 amino acids in length. The polypeptide predicted from the open reading frame would have a molecular weight of 57.4 kDa. Analysis of the amino acid sequence revealed an overall homology of 41.8% with the human adenovirus 2 (HAV-2) penton base. However, a region near the center of the polypeptide (amino acids 219 to 311) showed a significantly greater level of homology to the HAV-2 penton base (71%). Time course experiments using mRNA confirmed that this gene is expressed at late times postinfection. Upon probing genomic DNA from other FAV serotypes the penton base coding region hybridized to all six different serotypes tested, indicating a relatively conserved DNA sequence among a variety of FAVs.


Sequence analysis of the hepatitis B virus pre-C region in hepatocellular carcinoma [HCC] and nontumoral liver tissues from HCC patients. We investigated whether replication-competent pre-C/C defective mutants of hepatitis B virus (HBV) are detectable in primary human hepatocellular carcinoma (HCC) tissues from patients of a geographic area endemic for such mutants. DNAs extracted from formalin-fixed paraffin-embedded HCC samples were checked for the presence of specific HBV DNA sequences using the polymerase chain reaction (PCR). Amplified pre-C regions from nine HCC samples were directly sequenced as were samples of nontumoral liver tissues from five of these patients. The data show that hypervariable distal pre-C sequences were present in all nine HCC samples; this high variability was dependent on point mutations, which led to amino acid substitutions in nearly all cases. Interestingly, seven of the nine HBV DNA-positive samples from HCC tissues (but not samples from peritumoral liver tissue) showed mutations leading to amino acid substitution at the level of a distal cysteine residue. No mutation generating a translationally defective pre-C/C region was detectable in the tumor samples. Otherwise, in four of the six nontumoral liver tissues available from the same patients, a pre-C sequence with an in-frame TAG stop codon was detectable, although in three cases as a component of mixed population.


Phosphorylation of the Epstein-Barr virus BZLF1 immediate-early gene product ZEBRA. Expression of the Epstein-Barr virus (EBV) BZLF1 gene product ZEBRA is a first step in the cascade of the virus-productive cycle. ZEBRA protein was detected by immunoblotting as a single band at 38 kDa in Akata cells after crosslinkage of membrane immunoglobulin G (IgG) with anti-IgG antibody. Immunoprecipitation of [32P]phosphate-labeled, anti-IgG-stimulated Akata cells with anti-ZEBRA antibody showed that ZEBRA was phosphorylated. Phosphoamino acid analysis demonstrated phosphorylation of serine, but not threonine or tyrosine, and tryptic-peptide mapping showed multiple phosphorylated peptides of ZEBRA. Treatment with 8-bromo cAMP and blockage of phosphodiesterase by theophylline in anti-IgG-stimulated cells increased the phosphorylation of three ZEBRA peptides. Incubation with 12-O-tetradecanoylphorbol-13-acetate (TPA) reduced the phosphorylation of these three ZEBRA peptides, while treatment with staurosporine, a protein kinase C (PKC) inhibitor, enhanced their phosphorylations. These data suggest that activation of PKC with TPA induces the ZEBRA dephosphorylation and that activation of cAMP-dependent protein kinase A enhances the ZEBRA phosphorylation at the specific sites.


[Seasonal variations in nutrition behavior: results of the dietary survey of 1984/85 of the MONICA project in Augsburg]. Seven-day food records of the MONICA project Augsburg dietary survey, which were collected between October 1984 and May 1985 in 899 men aged 45-64 years, were used to analyse seasonal variations. Variations could be found in several food groups, but nutrient intake was nearly the same in autumn, winter and spring--with the exceptions of total fibre and alcohol.


A gene homologous to topoisomerase II in African swine fever virus. A putative topoisomerase II gene of African swine fever virus was mapped using a degenerate oligonucleotide probe derived from a region highly conserved in type II topoisomerases. The gene is located within EcoRI fragments P and H of the African swine fever virus genome. Sequencing of this region has revealed a long open reading frame, designated P1192R, encoding a protein of 1192 amino acids, with a predicted molecular weight of 135,543. Open reading frame P1192R is transcribed late after infection into a 4.6-kb RNA. The deduced amino acid sequence of this open reading frame shares significant similarity with topoisomerase II sequences from different sources, with percentages of identity between 23 and 29%. The evolutionary relationships among the topoisomerase II sequences of ASF virus, eukaryotes and prokaryotes were analyzed and a phylogenetic tree was established. The tree indicates that the ASF virus topoisomerase II gene was present in the virus genome before protozoa, yeasts, and metazoa diverged.


[The characteristics of craniocerebral trauma in diffuse toxic goiter]. The specific features of craniocerebral trauma in diffuse toxic goiter were studied from analysis of 34 cases. The craniocerebral trauma aggravated the course of diffuse toxic goiter with the development of thyreotoxic crises in the first days after the trauma in one of five patients. The therapeutic tactics in thyreotoxic crisis and the preoperative and postoperative management of patients are discussed.


[Work capacity status after acute craniocerebral trauma]. The working capacity of 1,855 patients after craniocerebral trauma (CCT) was analysed. A unified approach to the diagnosis of the type and severity of the CCT in all stages of the follow-up and to active complex treatment was recorded. The chances of restoration of working capacity are higher in young individuals, males, those engaged in intellectual work, and in persons with trauma suffered in everyday life. The possibility of disability is high in persons of prepension age, females, persons engaged in qualified physical work, and in trauma suffered at work. Medical factors, the severity of the trauma and reversibility of the deficiency signs have a more marked and significant effect on the condition of the working capacity at the outcome of CCT.


[The structural bases of the compensatory potentials of the central nervous system in closed craniocerebral trauma]. The article analyses 160 cases of fatal craniocerebral trauma encountered in the practice of the Burdenko Research Institute, AMS USSR, in 1980-1989. The results of the author's personal research and the literature data bear evidence of the different means of restoration of the functions of the central nervous system in localized and diffuse axonal brain damage. Reparative processes in the neurons of the pericontusion zone may provide compensatory changes. Compensation of disturbed functions of the central nervous system in diffuse axonal trauma is determined to a certain extent by the regenerative possibilities of partly damaged axons.


[The mechanisms of adaptation in severe trauma to the brain under conditions protecting it from hypoxia with pharmacological preparations and hyperbaric oxygenation]. From analysis of the adaptation mechanisms forming in severe brain trauma in 49 patients during its protection from hypoxia by combined administration of subnarcotic doses of sodium oxybutyrate and sodium thiopental in a bolus and an early course of hyperbaric oxygenation (HBO) the authors revealed the adaptation and disadaptation processes determining the outcome of the treatment. Since activation of the system of stress realizing biogenic amines promotes disadaptation processes, whatever the outcome, it is recommended to begin HBO after their stabilization. The character of changes of the lactate and pyruvate levels in blood flowing to and from the brain allowed the authors to distinguish the occurrence of a negative A-B gradient according to pyruvate after the first trial HBO session as a market of adaptation and a biochemical criterion of the expediency of prescribing a course of HBO.


Prenatal diagnosis of glucose-6-phosphate-dehydrogenase deficiency. Prior to the development of the DNA-based technology reliable prenatal diagnosis of G6PD deficiency was not possible. We show that, using PCR amplification and restriction endonuclease digestion, prenatal diagnosis is possible. We have now been able to determine that the male fetus of a mother heterozygous for G6PD Mediterranean had inherited the maternal X chromosome with the normal G6PD gene.


[The immunological aspect of the processes of adaptation and compensation in the acute period of craniocerebral trauma]. The dynamics of immunological values were studied in 350 patients with various types of craniocerebral trauma (CCT) in the acute period. It was established that immunological responses occur in all types of CCT. In mild CCT they are of a compensatory character. In severe trauma of the skull and brain the mechanisms of immunological protection are usually disrupted and secondary purulent complications and autoimmune processes develop. Under conditions of preliminary neurosensitization, the mechanisms of adaptation are disrupted even in mild traumas of the brain, autoimmune processes develop, which leads to a progredient course of CCT. Application of immunocorrective measures in disruption of adaptation processes improves the outcomes of CCT.


Neutrophil function in chronic neutrophilic leukemia: defective respiratory burst in response to phorbol esters. Functional analyses were performed on neutrophils isolated from 6 patients from two institutions who displayed features of chronic neutrophilic leukemia (CNL). These neutrophils demonstrated a consistent deficiency (44 +/- 8% of control values) in superoxide anion (O2-) production in response to the phorbol ester, phorbol myristate acetate (PMA). O2- production in response to chemotactic peptides was near normal (82.3 +/- 10.7% of control values). Bacterial killing was normal in the two patients studied, and chemotaxis was diminished in response to zymosan-activated plasma and to high concentrations of chemotactic peptides in the patients studied. Cytosolic C kinase activity was decreased in one of the two patients studied. These results suggest that a deficient O2- release in response to PMA is a hallmark of neutrophils in CNL and may provide a diagnostic indicator of this condition.


[The postaggressive reactions of the early period in acute craniocerebral trauma]. Experiments were conducted to study metabolic reactions developing in the first hours and days in response to craniocerebral trauma (CCT). Analysis of dynamics of the metabolism of glucose, lactate, and pyruvate, activity of enzymes in the blood, brain tissue, and in the cells, and the condition of the mitochondria made it possible to reveal the features of the course of postaggressive reactions in the early period of CCT. These reactions were hyperergic and their degree and trend depended on the severity of the brain damage.


[Prevention of pulmonary embolism].

[Prevention of pulmonary embolism]. Pulmonary embolism (PE) is one of the most common causes of death in hospitalized patients. Several groups of patients at high risk of developing venous thromboembolic complications have been identified. The effect of primary prophylaxis on the incidence of fatal PE has been demonstrated only in a few studies, while evidence for a benefit of pharmacologic and physical modalities of prophylaxis in preventing venous thrombosis has been documented in many prospective randomized clinical trials. The preventive measures differ according to the type of patient at risk, and in some groups of patients at risk several prophylactic regimen can be used. Application of low-dose heparin is the best documented prophylactic modality. General primary prophylaxis in 100 patients at risk could save one life. However, low-dose heparin is of limited efficacy in patients at high risk for thromboembolism. The following anithrombotic regimens are recommended in these patients: adjusted - dose heparin; oral anticoagulants with low intensity of anticoagulation; dextran; heparin in combination with dihydroergotamine; low-molecular-weight heparin fractions (LMWH). It is likely that prevention with LMWH will become the most important prophylactic modality in the near future. Mechanical methods of prophylaxis should be used primarily in combination with pharmacologic prevention. In low-risk patients mechanical methods alone offer sufficient prophylaxis. Low-dose heparin which is effective for primary prevention is not effective in secondary prophylaxis to prevent extension or embolization of established thrombosis. Inadequate anticoagulant therapy results in increased risk of recurrent thromboembolism. The general application of primary prophylaxis in patients at risk is superior to secondary prophylaxis with treatment of established thromboembolism.


[The adaptational reactions in mild experimental craniocerebral trauma and its combination with alcoholic intoxication]. The article deals with the results of experimental study of adaptation reactions in mild craniocerebral trauma (MCCT) combined with acute alcoholic intoxication on the basis of appraisal of the dynamics of endocrine and neuromediator shifts. A stress character of these shifts was revealed. Acute alcoholic intoxication levelled the stress reaction occurring in MCCT.


Neurological outcome in 22 treated adolescents with hyperphenylalaninemia. A clinical and electrophysiological study. We examined clinically and electrophysiologically 22 adolescents suffering from hyperphenylalaninemia. Three of 16 treated patients showed slight impairment of proprioception; their examination was otherwise unremarkable. Subclinical deficits were present in the central motor and sensory pathway. In addition, we had evidence of a minor sensory neuropathy: 32% of the 22 patients had subclinical deficits in the visual system. However, in contrast to earlier reports we saw no relation to the degree of metabolic control. Our results suggest that treated adolescent patients with hyperphenylalaninemia show neurological dysfunction that may be related to morphological, as well as pharmacological changes.


[The differentiated treatment of traumatic intracranial hematomas]. The article deals with the analysis of the clinical and computed tomography data, treatment, and outcomes in 94 patients with traumatic intracerebral hematomas (TICH). The indications for nonoperative treatment of TICH are a level of consciousness of the patient of no lower than 10 marks of the Glasgow coma scale, hematoma diameter of less than 4 cm, and the absence of clinical and computed tomography signs of brain stem compression. Osteoplastic trephination and encephalotomy is the principal surgical method. Puncture evacuation of the TICH may be resorted to if more than three fourths of its volume can be aspirated. The stereotaxic method is recommended for removal of TICH situated in the region of the basal ganglia.


Involvement of peripheral nervous system in juvenile Parkinson's disease. We evaluated, by using electrophysiological techniques, 29 patients with juvenile Parkinson's disease (JP), who had no known causes or clinical signs of neuropathy. Electromyographic evidence of chronic partial denervation with reinnervation was observed in nine patients (34.6%). Abnormalities of motor conduction in the common peroneal nerve were present in four (13.8%), Sural sensory conduction in nine (31.9%) and sympathetic skin response (SSR) in eleven (37.9%) patients. The symptoms of dysautonomia correlated poorly with changes in SSR. These abnormalities were independent of age at onset, duration or severity of the disease and antiparkinsonian drugs used. This study suggests that the peripheral nervous system is involved in JP in more than 50% of patients. The commonly observed symptoms of dysautonomia in Parkinson's disease may have a peripheral origin.


Natural course of acute and chronic monophasic inflammatory demyelinating polyneuropathies (IDP). A retrospective analysis of 266 cases. The present study analyses one of the largest IDP series published as yet: 266 cases, hospitalised between 1950 and 1983, conventionally treated and uniformly supervised. There was GBS in 84%, chronic IDP in 13%, polyneuritis cranialis in 1.5%, Miller Fisher and predominantly sensory neuropathy in 0.8% each. Numerous features including preceding events, course and outcome have been investigated. Latencies between onset and maximal deficit lacked (expected) bimodality. Sensory involvement was severe, mortality (18%) and maximal disability (20% requiring ventilation) high, autonomic dysfunction (71%) prominent. Statistical comparison of various features confirmed old age, rapid onset and need for ventilation as unfavorable predicting factors. However, preceding infection, bulbar paralysis and onset of paresis in proximal muscles evolved as unfavorable prognostic features as well.


Altered vasopressin response to metoclopramide in multiple system atrophy: evidence of a cholinergic defect in the hypothalamus. Multiple system atrophy (MSA) is a heterogeneous group of central neurological degenerations often associated with diffuse deterioration of the hypothalamic cholinergic neurons. In the hypothesis of an altered cholinergic regulation of vasopressin release, we evaluated vasopressin response to metoclopramide (20 mg i.v.), a cholinomimetic agonist, in 12 MSA patients. In the same patients the hemodynamic and osmolal control of vasopressin was also evaluated. We found that MSA patients had significantly lower basal plasma vasopressin values and higher plasma osmolality than control subjects. However, they displayed a normal vasopressin response to osmotic stimulation. During head-up tilting, orthostatic hypotension occurred in all patients, and the vasopressin response to hypotension was severely blunted in 5 of 12 patients, thus demonstrating the presence of a lesion of the afferent noradrenergic pathways. Metoclopramide increased vasopressin in control subjects, whereas MSA patients did not display any increase in vasopressin. These results clearly indicate that cholinergic neurons that regulate vasopressin release are damaged in MSA. Such an alteration may be dissociated from the lesion of the afferent noradrenergic pathways. As a consequence of the altered vasopressin release, MSA patients show lower plasma vasopressin levels with consequent propensity to dehydration and hypovolemia, which may further aggravate their hypotension.


Selective inhibition by dactinomycin of NANC sensory bronchoconstriction and [125I]NKA binding due to NK-2 receptor antagonism. In the present study, dactinomycin (10(-5) M) inhibited the non-adrenergic, non-cholinergic bronchoconstriction upon antidromic vagal nerve stimulation (1 Hz for 1 min) in the isolated perfused guinea-pig lung by 84%. The release of calcitonin gene-related peptide was unchanged, however, suggesting a postjunctional action. Dactinomycin (10(-5), 5 x 10(-5) M) also reduced non-adrenergic non-cholinergic bronchial contractions (maximally by 75%) induced by electrical field stimulation or capsaicin, while the cholinergic component and non-adrenergic non-cholinergic relaxation remained intact. The neurokinin-2 receptor antagonist L-659,877 (10(-6) M) had a similar effect as dactinomycin, inhibiting the non-adrenergic non-cholinergic bronchial contractions by 69%, while the neurokinin-1 receptor antagonist CP-96,345 (10(-6) M) had no effect. The bronchoconstriction evoked by neurokinin A, the selective neurokinin-2 receptor agonist Nle10neurokinin A (4-10) and capsaicin was markedly inhibited by dactinomycin while the contraction induced by substance P (SP), the selective neurokinin-1 receptor agonist Sar9Met(O2)11SP, endothelin-1 and acetylcholine was not affected. In autoradiographic experiments on guinea-pig lung, [125I]neurokinin A-labelled sections showed dense binding in the bronchial smooth muscle layer. Dactinomycin inhibited the specific binding of [125I]neurokinin A in a concentration-dependent manner (IC50 = 6.3 x 10(-6) M) and 66% of [125I]neurokinin A total binding was inhibited by 10(-4) M dactinomycin. In the rat colon, [125I]neurokinin A binding to neurokinin-2 sites on circular smooth muscle was inhibited by dactinomycin with an IC50 value of 7.9 x 10(-6) M. Dactinomycin failed to reduce increased nerve-evoked contractions or those caused by Nle10neurokinin A (4-10) per se in the rat vas deferens, which are considered to be mediated by neurokinin-2 receptor activation. In the rat portal vein, dactinomycin did not influence the contractions caused by the neurokinin-3 selective agonist Pro7neurokinin B. In conclusion, dactinomycin selectively inhibited neurokinin-2 receptor activation in guinea-pig lung and rat colon, but not in rat vas deferens, which may depend on the existence of different neurokinin-2 receptor subtypes. Neurokinin A is most likely the main endogenous excitatory non-adrenergic non-cholinergic transmitter in guinea-pig bronchi.


Na(+)-H+ exchange is not important for pancreatic HCO3- secretion in the pig. Pancreatic inter- and intralobular duct cells extrude H(+)-ions to interstitial fluid when they secrete HCO3- to pancreatic juice. This study assesses the potential importance of Na(+)-H(+)-ion exchange for H(+)-ion extrusion and secretion of HCO3-, using the Na(+)-H+ exchange blockers amiloride and hexamethylene-amiloride. Intracellular pH (pHi) in inter- and intralobular pancreatic duct epithelium was measured using BCECF fluorescence. H(+)-ion efflux was measured using a NH4Cl prepulse, acid-loading technique. In HCO3(-)-free media, pHi recovery following acid loading was blocked by amiloride (10(-4) M) and hexamethylene-amiloride (10(-6) M), demonstrating amiloride- and hexamethylene-amiloride-sensitive Na(+)-H+ exchange. However, 5 x 10(-6) M hexamethylene-amiloride did not reduce secretin-dependent pancreatic HCO3- secretion in vivo. Maximal H(+)-efflux through Na(+)-H+ exchange was 1.5 +/- 0.2 mumol min-1 ml cell volume-1, i.e. less than 1% of estimated net H(+)-ion efflux during HCO3- secretion. amiloride- and hexamethylene amiloride sensitive Na(+)-H+ exchange is not important for secretin-dependent pancreatic HCO3- secretion in the pig. Other mechanisms for H+ extrusion dominate.


High salt diet down-regulates proximal tubule Na+, K(+)-ATPase activity in Dahl salt-resistant but not in Dahl salt-sensitive rats: evidence of defective dopamine regulation. We examined the regulation of Na+,K(+)-ATPase activity in proximal tubule segments during a high salt diet in prehypertensive Dahl salt-sensitive and salt-resistant rats. Rats were placed on normal salt or high salt diets (0.9% saline as drinking water). During the normal salt diet, Na+,K(+)-ATPase activity was not different between Dahl salt-sensitive and salt-resistant rats. After 2 days and 10 days on a high salt diet, Na+,K(+)-ATPase activity in Dahl salt-resistant rats significantly decreased when compared to Dahl salt-resistant rats on a normal salt diet (P less than 0.01). The decreased Na+,K(+)-ATPase activity in Dahl salt-resistant rats during a high salt diet was reversed by treatment with an inhibitor of aromatic L-amino acid decarboxylase (dopamine synthesizing enzyme), benserazide. In contrast, Na+,K(+)-ATPase activity did not decrease during the high salt diet and benserazide had no effect on Na+,K(+)-ATPase activity in Dahl salt-sensitive rats. These results indicate that Dahl salt-sensitive rats do not have the capacity to down-regulate the proximal tubule Na+,K(+)-ATPase activity during a high salt diet. Indirect evidence suggests that the regulation of Na+,K(+)-ATPase activity by locally produced dopamine is absent in Dahl salt-sensitive rats.


Production of monoclonal antibodies against gastric parietal cell antigens. Two mice DBA/1 were each immunized with a single injection of one million enriched parietal cells in the hind foot pads. Monoclonal antibodies to be used as research tools in studies on regulatory mechanisms in gastric parietal cells were obtained after fusion of mouse myeloma cells (SP2) with cells from the popliteal lymph nodes of the mice. Twelve hybridomas produced antibodies reactive with structures only present in parietal cells as assessed by immunohistochemistry of oxyntic mucosa sections. Three hybridomas were subcloned and the antibodies produced by them, designated as PC4, PC8, and PC117, were characterized. In an enzyme-linked immunosorbent assay, all antibodies reacted with H,K-ATPase-containing vesicles. The antibody PC8 recognized a 94 kDa protein after immunoblotting of H,K-ATPase-containing vesicles and all antibodies precipitated a 94 kDa protein from [125I]H,K-ATPase-containing vesicles. The antibodies PC4 and PC117 recognized extracellular structures with a polarized distribution in viable, purified parietal cells. The results suggest that the structure recognized by all three antibodies is the alpha-subunit of the H,K-ATPase. The antibodies produced by another hybridoma, PC43, recognized a structure present in parietal and surface epithelial cells of the oxyntic mucosa. In an enzyme-linked immunosorbent assay, they reacted with a high-activity carbonic anhydrase which had been affinity-purified from pig oxyntic mucosa and they recognized a 30 kDa protein after immunoblotting. Thus, monoclonal antibodies against both intracellular and extracellular parietal cell structures were obtained after immunization with a small number of parietal cells.


Bisphosphonate space measurement in Paget's disease of bone treated with APD. The bisphosphonate space (BPS) is a quantitative measurement of skeletal uptake of 99mTc-HMDP. We measured BPS in 36 patients with Paget's disease of bone, both before and 6 months after treatment with intravenous APD (disodium pamidronate) infusions. BPS fell after treatment, but proportionally less than serum alkaline phosphatase (ALP) and fasting urinary hydroxyproline/creatinine (HYPRO). There was no dose-response relationship between the dose of APD given and the percentage reduction in ALP and HYPRO at 6 months. Log dose of APD/pretreatment BPS, however, predicted the percentage reduction in ALP and HYPRO very well, and from the respective regression equations it was possible to predict the dose of APD needed to achieve normal values of ALP and HYPRO. In the 10 patients who achieved a normal ALP and 9 patients a normal HYPRO after more than 6 months treatment with APD (range 7-18 months), the predicted dose of APD agreed closely with the actual dose. In conclusion, our data support the idea that log dose APD/pretreatment BPS is a valid predictor of biochemical response in Paget's disease.


Relation of client variables to continuance in five types of alcohol treatment settings. Multiple regression analyses were used to assess the relationship of 29 client variables to continuance in five types of alcohol treatment settings (outpatient, inpatient, social detoxification, halfway houses with highly structured treatment regimens, and halfway houses with minimal structured treatment; N = 3,240). Socioeconomic variables were most often related to continuance; however, the pattern of the relationship between predictor variables and continuance varied greatly from setting to setting. The strongest multiple regression coefficients were found for outpatient and inpatient settings and the weakest were found for the two types of halfway house settings. It was concluded that archive-derived variables had significant, but limited, power to predict continuance and that other approaches to investigating continuance are needed.


Personality characteristics of cocaine and alcohol abusers: more alike than different. Although there is extensive literature describing the characteristics of individuals entering treatment for alcoholism, few research reports are available describing the personality characteristics of individuals addicted to cocaine. Forty-four matched patient pairs, one member of each pair who abused cocaine and the other alcohol, were compared on the basis of their Minnesota Multiphasic Personality Inventory (MMPI) profiles. The subjects were matched on the basis of age, sex, and race from a larger population entering an inpatient addiction treatment center. Statistical analysis revealed more similarities than differences between the two groups, with the average profile type for both cocaine and alcohol abusers being 4-2. These results are discussed in the context of previous reports of psychopathology among cocaine abusers and personality theories of drug choice. The number of different code types within each substance abuse group is also reported and the concept of profile heterogeneity supported. Profile subtypes among cocaine patients are described clinically along with their implications for treatment.


Coincident Kaposi sarcoma and T-cell lymphoma in a patient with the Wiskott-Aldrich syndrome. A 24 year old male with a history of eczema, recurrent mild infections, and thrombocytopenia consistent with the Wiskott-Aldrich syndrome (WAS) presented with a mediastinal mass, generalized lymphadenopathy, splenomegaly, and severe thrombocytopenia. Studies of immune function including immunoglobulin levels and T-cell subsets were normal. Furthermore, his T lymphocytes proliferated normally in response to phytohemagglutinin, concanavalin A, and the combination of neuraminidase/galactose oxidase. However, their proliferative responses to anti-CD43 antibody and periodate were diminished, consistent with the clinical diagnosis of WAS. An initial inguinal lymph node biopsy surprisingly revealed Kaposi sarcoma. However, following splenectomy to increase the platelet count, biopsy of the mediastinal mass revealed T-cell large cell lymphoma. Studies of biopsied tissue for the presence of Epstein-Barr virus and cytomegalovirus were negative, as were studies of blood, including the polymerase chain reaction, for the presence of the human immunodeficiency virus (HIV). This is the first report of Kaposi sarcoma arising in a patient with a congenital immunodeficiency syndrome. Although Kaposi sarcoma can arise in the face of the severe immunosuppression that follows allograft transplantation and in patients infected with HIV, we postulate that longevity in the face of mild immunosuppression was the major factor in the development of Kaposi sarcoma in this patient.


Malignant mesothelioma in the jewelry industry. We conducted a clinical, environmental, pathologic, and mineral lung burden investigation of a 61-year-old man with malignant mesothelioma. For 35 years, up until three weeks prior to pneumonectomy, the patient made asbestos soldering forms at a costume jewelry production facility. Only chrysotile asbestos was used at the plant during the last decade of the patient's employment, and recent environmental sampling of the work-place identified no other asbestos fiber type. Anticipating that the patient would add to the very small number of cases of mesothelioma attributable solely to chrysotile, we found instead that the patient's lung tissue contained large numbers of both coated and uncoated amosite asbestos fibers but, surprisingly, no chrysotile. We subsequently learned that a distributor of both chrysotile and amosite supplied the company during the first 25 years the patient was fabricating soldering forms. The findings underscore the futility of estimating environmental exposure to chrysotile on the basis of fiber counts in lung tissue. Although we previously described non-neoplastic asbestos-related disease among patients engaged in similar work, this case, to the best of our knowledge, represents the first report of mesothelioma in the commercial jewelry industry. As such, it prompted us to initiate a public health campaign to replace asbestos soldering forms in this industry with readily available, safer alternatives.


Improving referral compliance after public cholesterol screening. Noncompliance with referral to a physician for retesting and diagnosis is a concern in public cholesterol screening. Participants (n = 2109) were referred by a health professional or lay communicator and randomly assigned to a coupon offer, referral reminder letter, or control group. A questionnaire was completed at screening, and a telephone interview was conducted 5 months later. Physician visit rates showed no professional or lay differences. For "no history" subjects, the behavioral interventions were effective compared with controls (coupon = 60.7% and reminder = 57.7% vs control = 46.1%). With professional counseling, only the coupon was effective; for lay counseling, both coupon and reminder yielded higher visit rates. Adjusted for sociodemographics, heart disease risk factors, and health perceptions, the intervention effects remained (professional-coupon offer: odds ratio [OR] = 1.94, 95% confidence interval [CI] = 1.21, 3.09; professional-reminder letter: OR = 1.04, 95% CI = 0.67, 1.63; lay-coupon offer: OR = 2.52, 95% CI = 1.52, 4.18; and lay-reminder letter: OR = 3.10, 95% CI = 1.83, 5.22). For unaware participants, lay counselors and referral follow-up efforts tailored to specific cholesterol risk groups are indicated.


Work-site nutrition intervention and employees' dietary habits: the Treatwell program. In a randomized, controlled study of the Treatwell work-site nutrition intervention program, which focused on promoting eating patterns low in fat and high in fiber, 16 work sites from Massachusetts and Rhode Island were recruited to participate and randomly assigned to either an intervention or a control condition. The intervention included direct education and environmental programming tailored to each work site; control work sites received no intervention. A cohort of workers randomly sampled from each site was surveyed both prior to and following the intervention. Dietary patterns were assessed using a semiquantitative food frequency questionnaire. Adjusting for work site, the decrease in mean dietary fat intake was 1.1% of total calories more in intervention sites than in control sites (P less than .005). Mean changes in dietary fiber intake between intervention and control sites did not differ. This study provides evidence that a work-site nutrition intervention program can effectively influence the dietary habits of workers.


Evaluation of an enzyme-linked immunosorbent assay for detection of antibodies to vesicular stomatitis virus in cattle in an enzootic region of Mexico. An ELISA was compared with the plaque-reduction serum neutralization (PRSN) test, for detection of vesicular stomatitis virus (VSV) antibodies in cattle in a vesicular stomatitis enzootic region of Mexico. A total of 325 bovine serum samples were screened for VSV antibodies. The PRSN test was performed, using Vero cells. The ELISA contained gradient-purified VSV Indiana (Lab strain) and VSV New Jersey (Hazelhurst) as the antigens. Regression analysis and weighted kappa statistic were used to estimate measures of agreement between the 2 assays for detection of VSV antibodies. The ELISA method proved useful for serodiagnosis of vesicular stomatitis. The ELISA and PRSN test results were highly correlated for detection of VSV antibodies.


Abortion induced by cell-associated pseudorabies virus in vaccinated sows. Pregnant sows, immune against pseudorabies after vaccination, were inoculated at 70 days of gestation either with autologous blood mononuclear cells that had been infected in vitro with pseudorabies virus (PRV) or with cell-free PRV. The infected cells or cell-free PRV were inoculated surgically into the arteria uterina. Eight sows (A to H) had been vaccinated with an inactivated vaccine. The titer of seroneutralizing antibodies in their serum varied between 12 and 48. Five sows (A to E) were inoculated with autologous mononuclear cells, infected either with a Belgian PRV field strain or with the Northern Ireland PRV strain NIA3. These 5 sows aborted their fetuses: 2 of them (B and C) 3 days after inoculation, and the other 3 (A, D, and E) 10, 11, and 12 days after inoculation, respectively. Sows F, G, and H were inoculated with a cell-free PRV field strain. They farrowed healthy litters after normal gestation. Neutralizing antibodies were absent against PRV in the sera of the newborn pigs, which were obtained prior to the uptake of colostrum. The 23 fetuses that were aborted in sows B and C 3 days after the inoculation were homogeneous in appearance and size. Foci of necrosis were not detected in the liver. Viral antigens were located by immunofluorescence in individual cells in lungs, liver, and spleen of 15 fetuses. Virus was isolated from the liver, lungs, or body fluids of 12 fetuses. The 39 fetuses that were aborted in sows A, D, and E between 10 and 12 days after inoculation were of 2 types: 17 were mummified and 22 were normal-appearing.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of initial restraint, weaning, and transport stress on baseline and ACTH-stimulated cortisol responses in beef calves of different genotypes. The productivity and well-being of animals can be substantially affected by stress. This is particularly true in the case of beef calves that are subjected to a multitude of stressors over a short period during the first year of life. Perhaps the most often studied stress-responsive variable has been blood corticosteroid concentrations. Factors such as age, gender, genetics, and degree of prior experience, can influence how an animal perceives and responds to a given stressor. Few studies have tried to control these variables, and accordingly, many conflicting results have been published regarding the impact of various stressors on cortisol response. We measured baseline plasma cortisol concentration over a 44-day study in Bos indicus and Bos taurus calves. Plasma cortisol values in Bos indicus calves were higher (32.60 +/- 0.66 ng/ml) than values in calves of Bos taurus (25.81 +/- 0.76) breeding. A precipitous decrease in cortisol concentration was observed 7 days after transport stress in all calves. Baseline cortisol concentration did not provide any indication of the intensity of the various stressors. However, significant differences were readily observed after ACTH administration. On the basis of cortisol secretion, stresses of transport and weaning were similar and were the most stressful to calves, regardless of genotype.


Incidence of localized feline leukemia virus infection in cats. Anecdotal descriptions of atypical FeLV infections, wherein standard clinical ELISA or immunofluorescence testing fails to detect active infections, suggest that an unknown proportion of FeLV-infected cats may go undetected. In this study, 127 viremic and nonviremic cats experimentally inoculated with FeLV were evaluated at necropsy for atypical expression of FeLV antigen. Results from viremic cats were in accordance with results of earlier studies on the pathogenesis of FeLV infection in cats, wherein antigen was found in lymphoid and epithelial tissues. Differences in time course or tissue distribution of viral antigen in some cats appeared to be attributable to the challenge virus preparations, consisting of cell-free tumor homogenate or infectious plasma. It was discovered that 5 of 19 of the FeLV challenge-exposed cats that were nonviremic had FeLV-specific antigens in select tissues (bone marrow, spleen, lymph node, and small intestine) 6 to 75 weeks after inoculation. These results indicated an additional category of possible outcomes for cats exposed to FeLV. Localized FeLV infection, as described here, may explain the discordance between clinical disease and laboratory testing for FeLV.


Ceruloplasmin and transferrin levels are altered in serum and bronchoalveolar lavage fluid of patients with the adult respiratory distress syndrome. The respiratory burst of neutrophils generates oxygen radicals that can result in lipid peroxidation and may contribute to acute lung injury in the adult respiratory distress syndrome (ARDS). Because ceruloplasmin and transferrin are inhibitors of lipid peroxidation and may play a role in regulating tissue injury, antigen levels of ceruloplasmin and transferrin and ceruloplasmin oxidase levels were measured in the serum and bronchoalveolar lavage fluid (BALF) of ARDS patients (n = 28), patients at risk for ARDS (n = 22), and normal control subjects (n = 45). Serum ceruloplasmin levels were similar in ARDS (mean +/- SEM) (3.8 +/- 0.3 microM) and at-risk (3.3 +/- 0.4 microM) patients compared with control subjects (3.2 +/- 0.2 microM). Serum transferrin levels were decreased in ARDS (14.9 +/- 1.7 microM) and at-risk (20.4 +/- 1.7 microM) patients compared with normal control subjects (32.9 +/- 1.2 microM), and serum transferrin levels correlated with serum unsaturated iron binding capacity (UIBC). Ceruloplasmin was detected in only one of 38 normal BALF samples (0.002 +/- 0.002 microM) and two of 13 at-risk BALF samples (0.15 +/- 0.1 microM), yet it was present in 17 of 23 ARDS BALF samples (0.9 +/- 0.2 microM). Transferrin was also increased in ARDS BALF (5.4 +/- 1.1 microM) compared with at-risk (0.7 +/- 0.5 microM) and normal (0.4 +/- 0.1 microM) samples. Ceruloplasmin that was present in the BALF and serum samples had functional oxidase activity, and purified human ceruloplasmin inhibited hydroxyl radical formation by phorbol myristate acetate (PMA)-stimulated neutrophils.(ABSTRACT TRUNCATED AT 250 WORDS)


Chest tumor response measurement during lung cancer chemotherapy. Comparison between computed tomography and standard roentgenography. Chemotherapy of lung cancer has, until now, been an experimental approach that requires careful evaluation of tumor response. The growing number of lung cancer patients now undergoing chemotherapy has led to a rapid increase in the number of computed tomography (CT) scans performed. Eighty consecutive lung cancer patients (55 non-small cell and 25 small cell lung cancers) were included in a prospective study to analyze whether the standard chest roentgenography is as effective as computed tomography in evaluating tumor response. Both standard chest roentgenography and CT scanning were performed before the chemotherapy began and were repeated after 10 to 12 wk of treatment. Response evaluations were performed according to the World Health Organization recommendations. When two-dimensional measurements were possible, the indicator lesions were defined as measurable tumors. Both roentgenography methods were used, independently, to classify the response into the following categories: complete response, partial response, minor response, stable disease, and progressive disease. A comparison of CT scans versus standard chest roentgenography as a measurement of indicator lesion showed a concordance of borderline significance (kappa = 0.146, p less than 0.05); a significant asymmetry was demonstrated (McNemar = 35.6, p less than 0.001), indicating that CT scanning may be a more appropriate method for measuring tumors than standard chest roentgenography. Moreover, no concordance was observed comparing CT scan and standard chest roentgenography measurability in the subgroups of patients with T3 or T4 tumor, hilar tumor, and patients with pleural effusion or atelectasis in which the McNemar test of symmetry constantly showed a better measurability using CT scan.(ABSTRACT TRUNCATED AT 250 WORDS)


There is no activation of O2- production by alveolar macrophages and neutrophil polymorphonuclear leukocytes in rat lung transplants during the reimplantation response and acute rejection. Activation of phagocytes (alveolar macrophages [AM] and neutrophil polymorphonuclear leukocytes [PMN]) can cause tissue damage in inflammatory lung diseases. In this study we investigated whether phagocytes contribute to the development of tissue damage in lung grafts histologically observed during two different processes: the reimplantation response and the acute rejection. Therefore, the number and profile of bronchoalveolar lavage (BAL) and blood phagocytes and their in vitro spontaneous and serum-treated-zymosan (STZ)-stimulated O2- production were assessed after allogeneic (BN to LEW) and syngeneic (LEW to LEW) transplantation of the left lung in rats. BAL PMN numbers increased during the reimplantation response, whereas during the late phase of the rejection process BAL AM and PMN numbers were increased. The O2- production by the BAL phagocytes and blood PMN were not increased at any stage. Strikingly, the STZ-stimulated O2- production by the BAL phagocytes was significantly impaired during acute rejection. Our data suggest that activation of the O2- production by bronchoalveolar phagocytes does not play an important role in the development of tissue damage in lung transplants during the reimplantation response and acute rejection. The impaired O2- production by alveolar phagocytes during acute lung rejection may contribute to the increased susceptibility for pulmonary infections after lung transplantation.


Activation of macrophages and cytotoxic cells during cytomegalovirus pneumonia complicating lung transplantations. The functional status of immune cells within human transplanted lungs was analyzed during cytomegalovirus (CMV) pneumonia complicating lung and heart-lung transplantations. The expression of interleukin-1 beta (IL-1 beta) and interleukin-6 (IL-6) genes is a marker for the activation of macrophages as is that of serine esterase B (SE-B) gene for cytotoxic cells. The levels of expression of these genes by bronchoalveolar lavage (BAL) cells were determined by in situ hybridization. Eight cases of CMV pneumonia were included in this study. BAL cells from either rejection episodes (eight cases) or control transplanted patients experiencing neither infection nor allograft rejection (eight cases) were analyzed in parallel. In the control patients, virtually no cells expressed the IL-1 beta, the IL-6, or the SE-B genes. In contrast, these three genes were all expressed in samples from patients with CMV pneumonia. IL-1 beta gene-expressing cells were abundant in all infected patients (mean +/- SEM: 898 +/- 449 positive cells per 10(4) cells, p less than 0.001, compared with those in control patients). IL-6 gene-expressing cells were less numerous (92 +/- 74 positive cells per 10(4) cells) and present in five of the eight cases of CMV pneumonia. Activated cytotoxic cells were detected in seven of the eight cases of CMV pneumonia (36.5 +/- 19 SE-B gene-expressing cells per 10(4) cells, p less than 0.001). During allograft rejections (eight cases) IL-1 beta gene-expressing cells were present in all but one patient.(ABSTRACT TRUNCATED AT 250 WORDS)


JC virus infection and Alzheimer's disease: reappraisal of an in situ hybridization approach. To assess the validity of the recently reported data on frequent occurrence of latent JC virus (JCV) infections in the brains of patients with Alzheimer's disease, we used in situ hybridization with biotinylated whole genomic JCV probes and the streptavidin-biotinylated alkaline phosphatase method to examine brain sections of such patients. We did not find any signs of JCV either in the brains of the patients with Alzheimer's disease or in those of nondemented, elderly control patients. Non-specific staining of corpora amylacea-like bodies, however, was invariably detected with in situ hybridization using JCV probes.


Localization of herpes simplex virus and varicella zoster virus DNA in human ganglia. Human dorsal root ganglia from 14 randomly autopsied adults and 1 infant (all seropositive for both herpes simplex virus [HSV] and varicella zoster virus [VZV]) were examined for latent HSV-1 and VZV DNA by polymerase chain reaction. Thoracic ganglionic DNA from all subjects and trigeminal ganglionic DNA from 11 adults were analyzed. HSV-1 DNA was detected in trigeminal ganglia from 8 of 11 (73%) adults and in thoracic ganglia from 2 of 14 (14%) adults. VZV DNA was detected in trigeminal ganglia from 10 of 11 (91%) adults and in thoracic ganglia from 12 of 14 (86%) adults. None of the DNA samples were positive with primers specific for HSV-2. These findings indicate the presence of latent HSV-1 and VZV DNA in trigeminal ganglia and latent VZV DNA in thoracic ganglia of most seropositive adults. Furthermore, although HSV-1 latency most commonly develops in trigeminal ganglia, we also show for the first time the presence of HSV-1 latency in thoracic ganglia. Finally, both viruses can become latent in the same trigeminal ganglion.